Western Regional Meeting Abstracts

Document technical information

Format pdf
Size 4.8 MB
First found Jun 9, 2017

Document content analysis

Language
English
Type
not defined
Concepts
no text concepts found

Persons

Organizations

Places

Transcript

WESTERN REGIONAL MEETING ABSTRACTS
Western Regional Meeting Abstracts
WAFMR, WSCI, WAP and WSPR
Joint Plenary Session II
1:45 PM
Thursday, January 27, 2011
1
INTRAUTERINE GROWTH RESTRICTION BY
UTEROPLACENTAL INSUFFICIENCY AND MATERNAL
TOBACCO SMOKING EXPOSURE CAUSES
GENDER-SPECIFIC CHANGES IN MRNA EXPRESSION OF
HIPPOCAMPAL ESTROGEN RECEPTOR
Numpang B, Caprau D, Ke X, Hale M, Yu X, Jiang C, Kaur R, Fitzhugh M,
Callaway C, Joss-Moore L, McKnight R, Lane RH University of Utah, Salt
Lake City, UT.
Purpose of Study: 2 major causes for intrauterine growth restriction
(IUGR) in the United States are uteroplacental insufficiency (UPI) and maternal tobacco smoking exposure (MTSE). Both UPI and MTSE impacts
neurodevelopmental outcomes in offspring born with IUGR, particularly in
males. A common characteristic of IUGR animal models is increased hippocampal apoptosis. Neuronal apoptosis is modulated by estrogen signalling estrogen is anti-apoptotic in the hippocampus. Estrogen mediates it effects
on hippocampal apoptosis through estrogen receptors (ER), which are encoded
by the genes esr1 and esr2. Currently, little is known about the effects of IUGR
by either UPI or MTSE on hippocampal expression of ERs.
Methods Used: IUGR was induced by 2 methods in 2 separate experimental groups: (1) uterine ligation in pregnant rats at day 19 of gestation to
mimic UPI, and (2) exposure of pregnant rats to tobacco smoke from day 11
of gestation to term to mimic MTSE. Both groups underwent caesarean
section at term. The pups were killed, hippocampi were dissected and RNA
extracted. Reverse transcriptase real-time PCR was performed to assay
expression of esr1 and esr2.
Summary of Results: UPI decreased esr1 (51% reduction, p=0.01) and
esr2 (55% reduction, p=0.02) expression in male pups compared to controls.
UPI did not alter esr1 or esr2 expression in female pups. In contrast, MTSE
increased esr2 expression (123% increase, p=0.05) in female pups compared
to controls, but no changes were seen in males. MTSE did not alter esr1
expression in either gender.
Conclusions: We conclude that the consequence of UPI and MTSE on
hippocampal ER mRNA expression is different, despite both UPI and MTSE
causing IUGR. The mechanisms behind MTSE induced IUGR is unknown,
but the impact is likely through MTSE induced fetal exposure to UPI, nicotine, and/or some other compounds. Given the different observed effects of
MTSE and UPI on hippocampal ER gene expression, we speculate that
mechanisms through which MTSE affects ER expression occur through
exposure to nicotine and/or some other compound.
2
MECHANICAL VENTILATION OF PRETERM LAMBS LEADS
TO MORE APOPTOSIS AND LESS PROLIFERATION OF
NEURONS AND GLIA
Alvord J, Wint A, McCoy MJ, Dong L, Dahl M, Callaway C, McKnight R,
Yoder BA, DiGernimo R, Lane RH, Albertine K University of Utah, Salt
Lake City, UT.
Purpose of Study: Prolonged mechanical ventilation (MV) of the premature human neonate is associated with lung injury. Frequently, the brain also
is injured. Our studies, using chronically ventilated preterm lambs, indicate
that MV changes histone marks in the brain. These changes could affect
regulation of cell processes, such as apoptosis and proliferation of neurons
and glia. The present studies purpose was to test this possibility. We
88
hypothesized that prolonged MV increases apoptosis and decreases proliferation among neurons and glia in the brain of preterm lambs.
Methods Used: Preterm (PT) lambs, treated with antenatal steroids and
postnatal surfactant,were managed by MVor high-frequency nasal ventilation
(HFNV; akin to bubble nasal CPAP (n=4 each) for either 3d or 21d. We use
HFNV as the positive gold-standard for alveolar formation in the lung. At the
end of 3d or 21d of ventilation, cortical brain tissue from the temporal lobe
was fixed and analyzed by double immunofluorescence to colocalize cleaved
caspase 3 or PCNA within neurons (neuronal marker-positive), immature
oligodendrocytes (O1-positive), or reactive astrocytes (GFAP-positive).
Summary of Results: Neurons in gray matter had significantly (*pG0.05)
more colocalization of cleaved caspase 3 in the MV group compared to the
HFNV group at 3d (22T3* vs 7T3/mm3) and at 21d (22T3* vs 11T4).
Conversely, neurons had significantly less colocalization of PCNA in the MV
group compared to the HFNV group at 3d (17T3* vs 23T7) and at 21d (27T7*
vs 16T1). Among glial cell types, the only colocalization difference was for
immature oligodendrocytes with cleaved caspase 3 at 21d. Significantly more
for immature oligodendrocytes were apoptotic in the MV group compared to
the HFNV group (29T3* vs 16T1).
Conclusions: We conclude that MV for 3d or 21d is associated with a shift
in balance between cell death and proliferation in the brain. The shift is
toward more apoptosis and less proliferation of neurons and immature oligodendrocytes. These results are consistent with less brain-derived neurotrophic factor in the same MV preterm lambs compared to We speculate that
imbalance results from epigenetic alterations in chromatin structure of genes
that regulate apoptosis and proliferation. (HL62875, HL56401, HD41075)
3
IDENTIFICATION OF A NOVEL BIOMARKER THAT
DISCRIMINATES HEART FAILURE ETIOLOGY
Maclellan WC, Maclellan R, Horwich T, Fonarow G, LeBlanc J UCLA, Los
Angeles, CA.
Purpose of Study: Despite improvements in the diagnosis and treatment of
CHF, these patients remain at high risk of cardiac mortality and ongoing
morbidity. Presently, the clinician_s ability to diagnose the etiology, determine prognosis or titrate therapy are limited. We sought to identify circulating
biomarkers that might address these limitations.
Methods Used: We analyzed plasma samples collected from 60 CHF
subjects through the Ahmanson-UCLA Cardiomyopathy Center and 44
normal subjects. Non-abundant plasma proteins were enriched by depletion
with an antibody-affinity column. Masses of the eluted plasma proteins were
determined by high-resolution MALDI-TOF mass spectroscopy.
Summary of Results: 794 proteins were detected within the plasma
samples by MALDI TOF-MS. 34 proteins were significantly different between the CHF patients and normal patients, using independent t-tests with a
PG0.05 and adjusted for multiple comparisons using the Bonferoni Correction. Significant proteins were then compared to a panel of clinical variables
(etiology, New York Heart Association (NYHA) classification, left ventricular ejection fraction (LVEF), and outcome to identify potentially important
biomarkers. This analysis demonstrated that the levels of 19 proteins correlated with etiology, 15 with LVEF, 2 with NYHA, and 2 with outcome. Three
of the unknown proteins were tentatively identified based on accurate mass
measurements (less than 0.5 Da error) as isoforms of APO C-III. To further
study APO C-III, we performed ELISA assays on 72 independent subjects.
The study population included 12 aged-matched normal subjects and 60 CHF
subjects, 30 dilated and 30 ischemic, all of them male, to measure circulating
apolipoproteinC-III levels in plasma. ApoC-III levels were elevated in subjects with dilated CHF compared to ischemic etiology and normals
(PG0.00001). ApoC-III levels remained significantly different (PG0.002) after
a multivariate analysis was to account for total cholesterol, total triglycerides
and age.
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Conclusions: We have used a novel, high through-put method to identify
potential biomarkers in plasma of CHF patients. Our initial results suggest
that ApoC-III levels correlate with CHF etiology. In the future, we will
confirm these findings in larger study populations.
4
RAPAMYCIN ADMINISTRATION IMPROVES METABOLIC
ABNORMALITIES AND DECREASES ADIPOSE TISSUE
INFLAMMATION IN DIET-INDUCED OBESITY
Subramanian S, Goodspeed L, Wang S, O’Brien KD, Chait A, Ladiges W
University of Washington, Seattle, WA.
Purpose of Study: Obesity is often associated with insulin resistance,
which may promote and increase the risk of cardiovascular disease. However,
the exact molecular mechanisms by which obesity leads to insulin resistance
are poorly understood. A hallmark of visceral obesity is the accumulation
of adipose tissue macrophages which can contribute to insulin resistance.
Aberrant, chronic activation of the intracellular nutrient sensing protein,
mammalian target of rapamycin (mTOR), a downstream effector of the
insulin signaling pathway, has been implicated in the pathogenesis of insulin
resistance. We hypothesized that oral administration of rapamycin, a potent
and specific inhibitor of mTOR, could reduce insulin resistance and improve
adipose tissue macrophage accumulation.
Methods Used: C57BL/6J mice were placed on a high fat, high carbohydrate Bdiabetogenic[ diet with rapamycin or placebo for 16 weeks. Rapamycin was administered admixed in the diet a microencapsulated form and
levels were detectable only in the drug-treated group.
Summary of Results: Mice that received rapamycin showed lesser weight
gain (pG0.01) compared to placebo fed animals despite equivalent food intake. Body composition analysis showed decreased body fat mass in the drug
treated group. Rapamycin-fed animals had lower plasma glucose (pG0.05)
and modest improvement in insulin sensitivity on insulin tolerance testing.
Plasma and hepatic triglyceride levels were reduced and morphologic analysis of the liver revealed decreased fat accumulation in hepatocytes in the
rapamycin group compared to the control animals. Immunohistochemical
analysis of perigonadal (visceral) adipose tissue showed decreased staining
for macrophage Mac2 in the mice treated with rapamycin. Gene expression
analysis of adipose tissue revealed decreased expression macrophage F4/80
and TNFa mRNA, while adiponectin gene expression was increased in the
rapamycin treated group.
Conclusions: Taken together, these data show that rapamycin improves the
metabolic profile and decreases adipose tissue inflammation in this mouse
model of diet-induced obesity. These results suggest that rapamycin and/or its
analogs have the potential for development as anti-obesity therapies.
WAFMR, WSCI, WAP, WSPR, WSMRF
Student Subspecialty Award Poster Session
6:00 PM
Thursday, January 27, 2011
5
BODY MASS INDEX AND ACUTE INJURY RISK IN
CHILDREN AGE 5-17 YEARS
McRae L1,2, Campbell J3, Doan Q2, Kissoon N2, AlQahtani A2 1University
of British Columbia, Vancouver, BC, Canada; 2BCCH, Vancouver, BC,
Canada and 3UBC, Vancouver, BC, Canada.
Purpose of Study: Obesity is an epidemic that affects many Canadian children. Between 1981 and 2001 the prevalence of overweight and obese
children in Canada increased by 200Y300%(1). In 2004, 18.1% of Canadian
children age 2Y17 were overweight and 8.2% were obese(1). Obese children
have prolonged recovery times and increased morbidity and mortality following an acute injury(2). The relationship between obesity and risk of acute
injury has not yet been established but is essential for proper weight loss
counseling of youth. Our study investigates the risk of acute injury associated
with increasing body mass index (BMI) in children.
Methods Used: This is a case based, case control study with patients_ age 5
to 17 years who presented to BC Children_s Hospital Emergency Department
from July 2009 to December 2010. Cases were defined as children who
presented for assessment of an injury. Controls were children with any other
complaint who had not sustained an injury in the previous 12 months. Height
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
and weight were measured and BMI categories (underweight, normal, overweight and obese) were assigned according to the CDC classification per age
and sex. Descriptive statistics were used to report distribution of participants_
characteristics (BMI, sex, age, physical activity, socioeconomic status).
Multivariate logistical regressions estimated the odds of injury occurrence by
BMI category adjusted for confounders.
Summary of Results: To date we have screened 500 patients and 394 were
enrolled. Although the proportion of male subjects (45%) and BMI category
distribution (22Y23% overweight and obese combined) was similar among
cases and controls, the distribution for the other variables were considerably
distinctive. The control population is younger, less active and less affluent.
Results of logistic regression analyses are pending.
Conclusions: Although an increased risk of acute injury in overweight and
obese children cannot be concluded from our descriptive analyses, a full
analysis is pending. 1. Starky S. The obesity epidemic in Canada. {http://
www2.parl.gc.ca/Content/LOP/ResearchPublications/prb0511-e.htm#
introductiontxt} (Accessed Sept 23, 2010). 2. Pomerantz W, Timm N,
Gittelman M. Injury patterns in obese versus nonobese children presenting
to a pediatric emergency department. Pediatrics 2010;125(4):681-85.
6
EXAMINATION OF VERBAL INTELLIGENCE AND
PHONOLOGICAL PROCESSING IN CHILDREN WITH
AUTISM SPECTRUM DISORDERS AND THEIR
FAMILY MEMBERS
Thomson AL1, Webb SJ2, Bernier R2, Wijsman E2 1University of Washington
School of Medicine, Seattle, WA and 2University of Washington, Seattle, WA.
Purpose of Study: Autism Spectrum Disorders (ASD) have a strong
genetic influence, and a broader autism phenotype (BAP) has been observed
in first-degree family members. It is well known that children with autism
have an array of communication deficits, including phonology, semantics,
syntax, and pragmatics. The overall aim of this project was to better understand verbal language use in individuals with ASD and their family members.
Methods Used: A total of 279 family members from families with 2 or more
ASD children participated. Verbal intelligence quotients (VIQ), a measure of
vocabulary and reasoning, were assessed with age-appropriate Wechsler
scales. Non-word repetition (NWR), a measure of phonological processing,
was assessed with the CTOPP.
Summary of Results: As expected, ASD children exhibited lower VIQ
compared to mothers, fathers, and unaffected siblings (F(3,274) = 59.8,
p G.01).To examine decreased VIQ in the context of overall cognitive function, discrepancy scores were calculated between verbal and non-verbal IQ.
ASD children_s IQ discrepancy scores were significantly greater than relatives (F(3,274) = 19.8, p G.01). Both our higher-functioning (HF) and lowerfunctioning (LF) ASD groups exhibited a mean discrepancy pattern that
suggested better non-verbal IQ compared to VIQ (pG.05). For phonological
processing, mean performance for all groups was significantly below the
norm on the NWR task, however siblings performed relatively better than
ASD children and fathers (F(3,256) = 2.3, pG.05). Investigation of the relationship between VIQ and NWR revealed significantly higher ratio scores in
ASD children and siblings compared to parents (F(3,253)=11.976, p G.01).
Conclusions: These results suggest that relatively poorer phonological
processing may be an endophenotype of ASD and a part of the BAP. The ratio
of the phonological processing to VIQ suggests that the relation between
these two domains changes with age, but not affected status. Taken together,
these data suggest that multiple aspects of language processing contribute to
both autism and the broader phenotype and will be important quantitative
traits for genetic analyses.
7
INCIDENCE, ETIOLOGY, AND SURVIVAL TRENDS FROM
SUDDEN CARDIAC ARREST IN CHILDREN AND YOUNG
ADULTS AGE 0-35 IN KING COUNTY, WASHINGTON:
A 30-YEAR REVIEW
Meyer L1, Drezner J1, Fahrenbruch C2, Stubbs B2, Maeda C1, Harmon K1,
Eisenberg M1,2 1University of Washington, Seattle, WA and 2King County
Emergency Medical Services, Seattle, WA.
Purpose of Study: Sudden cardiac arrest is a leading cause of death in
children and young adults, although little is known regarding the epidemiology
89
Western Regional Meeting Abstracts
Journal of Investigative Medicine
of such events. This study determined the incidence, etiology, and outcomes
of out-of-hospital cardiac arrest (OHCA) in individuals age 0Y35.
Methods Used: This was a retrospective cohort study of OHCA in children
and young adults from January 1980 through December 2009 in King
County, Washington. Cases of OHCA were identified from the King County
Public Health Division of Emergency Medical Services (EMS) Cardiac
Arrest Database, an ongoing registry of all cases of OHCA involving an EMS
response. Incidence rates were calculated using population census data, and
the etiology of cardiac arrest determined by review of autopsy reports, death
certificates, hospital and other available records.
Summary of Results: A total of 361 cases (26 cases age 0Y2, 30 cases age
3Y13, 60 cases age 14Y24, and 245 cases age 25Y35) of OHCA occurred,
with an overall incidence of 2.28 per 100,000 person-years (2.1 in age 0Y2,
0.61 in age 3Y13, 1.44 in age 14Y24, and 4.40 in age 25Y35). The most common etiologies of OHCA were congenital abnormalities (84.0%) in age 0Y2,
hypertrophic cardiomyopathy (17.9%) in age 3Y13, presumed primary arrhythmia (23.5%) in age 14Y24, and coronary artery disease (42.9%) in
age 25Y35. The overall survival rate was 26.9% (3.8% in age 0Y2, 40.0% in
age 3Y13, 36.7% in age 14Y24, and 27.8% in age 25Y35). Survival increased
significantly throughout the study time period from 13.0% in 1980Y1989 to
40.2% in 2000Y2009 (pG0.001).
Conclusions: The incidence of OHCA in children and young adults is
substantially higher than previously reported. This study details the various
cardiac disorders leading to OHCA in the young, allowing an increased focus
on age related risk factors and development of effective screening models
for prevention. This study demonstrates an important increase in survival,
providing compelling support of contemporary resuscitation protocols for
OHCA in the young.
8
PEDIATRIC MALARIA TREATMENT DELAYS ACROSS
SOCIOECONOMIC LEVELS IN SUBURBAN ACCRA, GHANA
Wong KR 1Yale University School of Medicine, New Haven, CT; 2University
of California at San Diego, La Jolla, CA and 3 University of Ghana School
of Public Health, Accra, Ghana.
Purpose of Study: The study reported here examines how a Ghanaian
family_s socioeconomic status (SES) affects the delay-time before a child
with Plasmodium falciparum malaria is brought to a government hospital. It
was hypothesized that a delay of appropriate clinical treatment results from a
family_s apprehension of inability to pay the hospital fee.
Methods Used: In this ethnographic study, parents of pediatric patients with
blood smear-confirmed malaria (N=15) were interviewed through both
quantitative questionnaires and narratives at the Legon Hospital Pediatrics
Ward. The questionnaires ascertained the dependent variable (time lag during
which the child languished at home), as well as the independent variable
of SES, measured by: 1) monthly income range, 2) health insurance, and 3)
years of household education. Over the course of four months, 170 hours of
dialogue and observation supplemented the numerical data.
Summary of Results: There was actually no statistical association between
delay-time and family financial standing, nor a correlation between delaytime and enrollment in Ghana_s recently-established National Health Insurance Scheme. Parents of all backgrounds postpone clinical treatment even
after the onset of clear malaria symptoms in order to conserve their time;
the waiting lines at hospitals are often several hours long and the illness
is endemic in West Africa. Yet, this study does confirm that lower-SES
households less effectively prevent malaria. Only half of families in the
lowest income group utilized insecticide-treated mosquito nets, while all six
families in the highest income group use the precaution.
Conclusions: In this township, the study suggests that low-SES families do
not disproportionately delay clinical treatment. Especially when raising
multiple children, parents of all SES groups delay at home and self-treat with
herbal teas or street-bought (and often counterfeit) chloroquine and artesunate anti-malarials. This has its risks. Delayed clinical care and the rapid
deterioration of originally-uncomplicated P. falciparum cases contribute to
20,000 pediatric malaria fatalities in Ghana annually.
9
TISSUE SPECIFIC ROLE OF DNA METHYLTRANSFERASE
4 DURING ZEBRAFISH DEVELOPMENT
Hooton J1, Jafri I2, Jones DA2 1University of Washington School of
Medicine, Seattle, WA and 2University of Utah, Salt Lake City, UT.
90
&
Volume 59, Number 1, January 2011
Purpose of Study: DNA methylation is a heritable epigenetic marker. The
mechanism by which DNA methylation and gene expression are regulated
during embryonic development is an area of active research. The zebrafish
presents a unique model which is well suited to investigate mechanisms of
normal embryonic development because of its easily observed rapid embryonic development and availability of tools to manipulate gene expression.
Recent research has started to establish tissue specific roles for the various
DNA methyltransferases (DNMTs) during development. These studies have
also shed some light on the basic mechanisms by which DNMTs regulate
expression of targeted genes. DNMT3 is one of the three groups of DNMTs
found in humans and it has six known zebrafish orthologs: Dnmt3/4/5/6/7/8.
The focus of this project was to investigate the role of zebrafish dnmt4 in
normal development. Whole mount in situ hybridization assays revealed that
dnmt4 is robustly expressed in hematopoietic compartments as well as the
site of hematopoietic stem cell generation. We hypothesized that dnmt4
controls zebrafish hematopoiesis by regulating the transcription of hematopoietic genes.
Methods Used: We took a candidate approach and investigated whether the
knockdown of dnmt4 caused transcriptional up regulation of aid, mbd4, and
gadd45a. These targets were chosen based on recent findings both in published studies and our lab_s unpublished work that indicates they play a role in
hematopoiesis. Antisense morpholino knockdown technology was used to
create zebrafish embryos with substantially decreased dnmt4 expression.
RNA was isolated from whole fish samples and then cDNA was created. RTPCR was conducted to test the relative expression normalized to 28s of the
target genes.
Summary of Results: Expression of our target genes was found to be two
to three times higher in dnmt4 morphants than in controls.
Conclusions: Taken together these data provide evidence that dnmt4 plays a
role in hematopoietic differentiation and establishes a possible tissue specific
role for dnmt4 during embryonic development. Future studies will be aimed
at further characterizing the hematopoetic phenotype of dnmt4 morphants
and verifying that changes in gene expression correlate with tissue specific
changes in DNA methylation of target genes.
10
ESTRADIOL17A (E2) ATTENUATES THE GNRH MEDIATED
INCREASE IN INTRACELLULAR CA2+
Mrdutt M1, Magee C2, Cantlon J2, Murtazina D2, Sanborn B2, Clay C2
1
Univ of Washington, Seattle, WA and 2Colorado State Univ., Fort Collins, CO.
Purpose of Study: Controlling the ovulatory LH surge underlies fertility
regulation. The LH surge is initiated by E2 actions at the hypothalamus and
the pituitary to increase GnRH receptor expression on gonadotropes - the LH
producing cells. Interestingly, however, E2 acutely but transiently suppresses
GnRH mediated LH release thus increasing pituitary content of LH just prior
to the surge. We hypothesized that E2 acutely attenuates the GnRH elicited
increase in intracellular Ca2+ - the cellular event underlying GnRH mediated
LH secretion.
Methods Used: We established cultures of pituitary cells from transgenic
mice in which expression of a fluorescent protein (YFP) is confined to
gonadotropes. The distinct excitation spectra of YFP and the Ca2+ indicator
FURA allowed for selection of gonadotropes and subsequent imaging of
GnRH induced changes in intracellular Ca2+. After 18 hr in culture, cells
were Bloaded[ with FURA for 1 hr. Cells were then incubated in Ca2+
containing or Ca2+ free media and received 100nM E2 or vehicle followed by
a 2nM GnRH pulse at 2 and 12 min post-E2. Increases in intracellular Ca2+
were determined by a change in FURA emission spectra. Both amplitude
(peak height) and magnitude (area under curve) of the Ca2+ response in 136
cells was analyzed 1 min post-GnRH.
Summary of Results: Approximately 93% of YFP positive cells displayed
a Ca2+ response following the initial GnRH pulse. Of these, 84% displayed a
secondary Ca2+ response; however, in both Ca2+ and Ca2+ free conditions,
the magnitude and amplitude of the secondary response was reduced
(pG0.05) as compared to the initial response. E2 did not alter the percentage
of cells responding to either GnRH pulse. However, in both Ca2+ and Ca2+
free conditions, E2 further reduced (pG0.05) the ratio of secondary versus
primary response magnitude by 25% and 31%, respectively.
Conclusions: Consistent with our hypothesis, E2 attenuates the GnRH induced Ca2+ response in gonadotropes - a potential explanation for the acute
block of GnRH mediated LH release by E2 prior to the ovulatory LH surge.
The rapidity of the inhibitory effect of E2 suggests a non-nuclear mechanism
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
of action. If correct, new generations of E2 agonists/antagonists may allow
for fertility regulation without the myriad physiological effects resulting
from nuclear actions of E2.
11
WILSON DISEASE: ABNORMAL METHIONINE
METABOLISM IN PATHOGENESIS OF LIVER DAMAGE
Dolatshahi M, Jose S, Halsted C, Medici V University of California, Davis,
Davis, CA.
Purpose of Study: Wilson disease (WD) is a recessive disorder of copper
accumulation with established genetic background but unclear pathogenesis.
Aberrant hepatic methionine metabolism plays a role in the pathogenesis of
liver injury by regulating homocysteine (HCY) and S-adenosylhomocysteine
(SAH), which induce endoplasmic reticulum (ER) stress and consequent
induction of lipogenic pathways. We hypothesized that abnormal methionine
metabolism is involved in the pathogenesis of hepatic steatosis in WD.
Methods Used: Livers were removed from Jackson toxic milk mice (tx-j), a
model of WD, and C3H control mice fed the same diet at 16, 20, and
24 weeks of age. Profiles of selected hepatic methionine metabolites in the
tx-j mice were compared to those of the wild type mice at the three ages and
were correlated with histopathology, triglyceride accumulation, and activation of ER stress, apoptosis, and lipogenesis markers.
Summary of Results: At 20 and 24 weeks, the tx-j mice showed greater
hepatic triglyceride levels and higher cytoplasmic lipid droplets in hepatocytes compared to the controls. SAH hydrolase (SAHH) mRNA expression,
protein levels, and activity were significantly lower in the tx-j mice at all ages
and correlated negatively with hepatic copper levels, which were significantly
higher in all tx-j mouse age groups. The transcript and protein levels of the
ER stress marker glucose related protein 78 (GRP78), the apoptosis marker
growth arrest DNA damage inducible gene 153 (GADD 153), and the lipogenesis transcription factor sterol regulatory element binding protein 1c
(SREBP-1c) were increased at all age points in the tx-j mice. Plasma HCY
levels were elevated at 20 weeks and correlated with hepatic triglyceride
levels.
Conclusions: Reduced hepatic SAHH expression was associated with increased SAH and subsequently with increased plasma HCY as well as the
activation of ER stress lipogenic pathway gene expression, which were paralleled by the progression of hepatic damage. These findings support the
hypothesis that aberrant methionine metabolism plays an important role in
liver damage pathogenesis in WD.
12
VITAMIN D INHIBITS PROLIFERATION AND INDUCES
DIFFERENTIATION OF C2C12 SKELETAL MYOBLAST
CELLS BY MODULATING THE EXPRESSION OF
PRO-MYOGENIC AND ANGIOGENIC GROWTH FACTORS
King KK1,2, Garcia LA1, Ferrini M1,2, Norris KC1,2, Artaza JN1,2 1Charles
R. Drew University of Medicine and Science, Los Angeles, CA and 2David
Geffen Medical School at UCLA, Los Angeles, CA.
Purpose of Study: Skeletal muscle wasting is a serious public health
problem associated with aging, chronic disease, kidney dialysis and AIDS.
Vitamin D (VD) is widely recognized for its regulation of calcium and
phosphate homeostasis in relation to bone development and maintenance and
for its synergistic effects on target organs such as intestines, kidneys and PTH
glands. It has been shown to improve muscle performance and reduce falls in
VD deficient older adults. Little is known of the underlying mechanism or
role it plays in association with myogenic differentiation. In this study, we
examined the effect of 1,25-D -the active form of VD- at the molecular level
on myoblast cell proliferation, progression and differentiation into myotubes.
Methods Used: C2C12 myoblasts were treated with and without 1,25D in a
time course manner. VDR expression was analyzed by: Immunofluorescence
(IF), Real Time PCR (qRT-PCR) and Western Blots. Expression of muscle
lineage and proliferation markers was assessed by immunocytochemistry (ICC)
and western blots. Expression of pro-myogenic markers and angiogenic growth
factors was analyzed by qRT-PCR arrays and confirmed by qRT-PCR.
Summary of Results: Addition of 1,25D to C2C12 myoblast induces: a)
increase expression and nuclear localization of the VDR; b) decrease PCNA
(a cell proliferation marker) cell expression; c) increase expression of MyoD
and myogenin (early and intermediate myogenic markers respectively); d)
increase expression of IGF-2, FST (follistatin-a myostatin inhibitor), FGF1
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
(fibroblast growth factor 1, involved in muscle development and regeneration) and VEGF which has been demonstrated to promote myotube hypertrophy and increased myogenic differentiation; e) no changes in GDF11
expression; and f) decrease expression of IGF-1 and Mstn- the only known
negative regulator of muscle mass.
Conclusions: This study provides a mechanistic justification for VD
replenishment in muscle waste conditions such as AIDS, cancer, congestive
heart failure, COPD, renal failure, and also in VD deficient older adults who
are known to have age-related loss of muscle mass and strength and an
increased rate of falls.
13
RISK FACTORS FOR EPILEPSY IN TERM INFANTS WITH
HYPOXIC ISCHEMIC ENCEPHALOPATHY
Hong KJ1, Bonifacio SL3, Barkovich AJ4, Sullivan JE2,3, Rogers EE3,
Ferriero DM2,3, Glass HC2,3 1University of Washington, Seattle, WA;
2
University of California San Francisco, San Francisco, CA; 3University of
California San Francisco, San Francisco, CA and 4University of California
San Francisco, San Francisco, CA.
Purpose of Study: Hypoxic ischemic encephalopathy (HIE) is a significant
cause of neonatal death and adverse neurodevelopmental outcome, such as
cerebral palsy, developmental delay, and epilepsy. The goal of this study was
to determine neonatal predictors of epilepsy in term newborns with HIE using
clinical data and MRI.
Methods Used: This was a cohort study of 195 term infants with birth
asphyxia, born 1993 through 2009. Neonatal clinical data, such as neonatal
seizures, encephalopathy scores, and MRI pattern of injury, were obtained
through chart review. Parents of the infants were contacted by telephone and
administered a seizure questionnaire. Neurodevelopmental data including
cognitive scores, neuromotor scores (NMS), and neurological outcomes were
extracted from standardized longitudinal follow up data.
Summary of Results: Of 121 children contacted, 12 (10%) developed
epilepsy. Univariate analyses showed significant differences between epilepsy
and no epilepsy groups for neonatal seizures, EEG seizures, status epilepticus, encephalopathy scores of 5 or 6, and brain injury on MRI (PG0.01). Of
49 infants with neonatal seizures, 12 (24%) developed epilepsy. The infants
with EEG seizures (RR=9; 95% CI=3Y27) and status epilepticus (RR=11;
95% CI=5Y25) were more likely to develop epilepsy than those without.
There was no significant association for clinical seizures (P=0.5) or encephalopathy scores of 3 or 4 (P=0.1) and epilepsy. The children with epilepsy had injury on MRI: 5 had basal ganglia patterns and 7 had watershed
patterns. Children with epilepsy had an average cognitive score of 54T11,
whereas children with no epilepsy had a score of 97T17 (PG0.005). Median
(range) NMS were 5 (0Y5) for children with epilepsy and 0 (0Y5) for children
with no epilepsy (PG0.005).
Conclusions: Term infants with HIE and neonatal seizures, specifically
electrographic seizures and status epilepticus, evidence of brain injury on
MRI, and more severe encephalopathy scores are at a higher risk of developing epilepsy. Our data provide information for clinicians to use when
counseling parents on the possible outcome of epilepsy.
14
INSULIN-LIKE GROWTH FACTOR II PEPTIDE FUSION
ENABLES UPTAKE OF ALPHA-N-ACETYLGLUCOSAMINIDASE
INTO MUCOPOLYSACCHARIDOSIS TYPE III B FIBROBLASTS
Sinow C, Kan S, Tippin B, Troitskaya L, DiStefano A, Todd A, Le S,
Dickson P LA BioMed at Harbor-UCLA, Torrance, CA.
Purpose of Study: Mucopolysaccharidosis type IIIB (MPS IIIB; Sanfilippo
B) is a lysosomal storage disorder characterized by deficiency of >-N-acetylglucosaminidase (NAGLU) causing defective degradation and accumulation
of heparan sulfate. Enzyme replacement therapy (ERT) has been used to treat
lysosomal storage diseases but has not been successful for MPS IIIB. In
particular, ERT for MPS IIIB has been limited by inadequate cellular uptake
of recombinantly produced human NAGLU enzyme. In this study we create,
characterize, and test a novel enzyme: NAGLU fused to the insulin-like
growth factor 2 (IGF-II) peptide (rhNAGLU-IGF-II). The IGF-II motif was
selected to enable recombinant NAGLU to enter cells via the insulin-like
growth factor 2 binding site found on the mannose 6-phosphate receptor for
high affinity delivery to lysosomes.
91
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Methods Used: Following the molecular cloning of the recombinant
NAGLU-IGF-II construct, rhNAGLU-IGF-II was stably expressed in
Chinese hamster ovary cells in secreted form and purified. Activity assays,
biochemical analysis and substrate kinetics were performed. Cellular uptake
by human MPS IIIB fibroblasts and inhibition assays were followed by
confocal microscopy to evaluate the subcellular distribution of rhNAGLUIGF-II in the fibroblasts. Glycosaminoglycan (GAG) storage reduction
was measured.
Summary of Results: The enzyme activity towards the substrate 4-MUNG
was comparable to wild-type rhNAGLU, with peak activation near lysosomal
pH. MPS IIIB fibroblasts and brain-derived cell lines readily took up the
fusion enzyme via receptor-mediated endocytosis that was inhibited by
competitive IGF-II peptide. Co-localization of rhNAGLU-IGF-II enzyme
with lysotracker red in MPS IIIB cells via confocal microscopy confirmed
lysosomal targeting. GAG storage was also reduced up to 50% (pG0.001) in
MPS IIIB cells after treatment with rhNAGLU-IGF-II when measured by
radioactive incorporation assay.
Conclusions: Our data suggest that the fusion enzyme, rhNAGLU-IGF-II,
can enter human cells and localize in the lysosomes. Future studies in vivo are
needed to more fully assess the clinical potential of rhNAGLU-IGF-II as
an ERT for Sanfilippo B syndrome.
15
BONE SCAN OVERUSE IN THE STAGING OF
PROSTATE CANCER
Palvolgyi R1, Daskivich T1, Kwan L1,3, Chamie K1,3, Litwin M1,2,3
1
UCLA David Geffen School of Medicine, Los Angeles, CA; 2UCLA School
of Public Health, Los Angeles, CA and 3UCLA, Los Angeles, CA.
Purpose of Study: With contemporary widespread PSA screening, more
patients exhibit low- and intermediate-risk prostate cancer, thus reducing the
need and yield of bone scan imaging. We aimed to determine the utilization
and subsequent positivity rates of bone scan imaging in a contemporary
Veterans Affairs (VA) cohort of men with prostate cancer.
Methods Used: We retrospectively reviewed the charts of 1597 men
diagnosed with prostate cancer between 1997 and 2004 at the Greater Los
Angeles and Long Beach VA Medical Centers. We used univariate analysis to
measure association between patient (age, race, comorbidity) and tumor
(PSA, clinical stage, Gleason score) characteristics with bone scan use and
positivity. We conducted the analysis for scans that were and were not clinically indicated, according to established AUA guidelines.
Summary of Results: Out of 1121 men not indicated for nuclear imaging,
452 (40%) received a bone scan with only 6 (1%) returning positive. Among
the same group, increasing PSA, clinical stage, Gleason score, and subsequent D_Amico tumor risk, were all positively associated with bone scan
overuse, but not with corresponding positivity rates. Univariate analysis
showed age, PSA, clinical stage, Gleason score, and treatment type as significant predictors of bone scan overuse.
Conclusions: Bone scan overutilization in men with clinically localized
prostate cancer results in unnecessary patient anxiety, time consumption, and
significant economic waste for the VA. If the 40% overutilization rate for
clinically localized prostate cancer reflects national overuse patterns, then
implementing a quality improvement initiative to minimize overutilization
would translate an annual savings of $9,409,680 for the VA, without compromising oncologic outcomes.
92
&
Volume 59, Number 1, January 2011
16
YIN YANG 1 REGULATES THE TRANSCRIPTIONAL
REPRESSION OF SURVIVIN
Galloway N1, Li F2, Sui G3, Wall N1 1Loma Linda University School of
Medicine, Loma Linda, CA; 2Roswell Park Cancer Institute, Buffalo, NY
and 3Wake Forest University School of Medicine, Winston-Salem, NC.
Purpose of Study: Survivin is a member of the Inhibitor of Apoptosis
(IAP) family of proteins, and is highly expressed in all cancers but absent in
normal tissue. Expression level correlates with chemo- and radioresistance,
as well as poor prognosis in cancer patients. The mechanisms for upregulation of Survivin in cells undergoing stress associated with tumor development
and the tumor microenvironment are not well understood. The putative stress
response transcription factor Yin Yang 1 (YY1) was hypothesized to contribute to the upregulation of survivin in tumor cells.
Methods Used: In order to study regulation with luciferase reporter assays,
U2OS cells were transfected with nested deletions of the survivin promoter,
ranging from short (+230 bp) to long (+6280 bp). YY1 involvement in
survivin promoter repression was confirmed using siRNA directed against
YY1. A U2OS cell line containing a stable YY1 Tet-off system was used to
determine whether a temporal increase in YY1 expression affects Survivin
protein levels. To further evaluate the role of YY1 regulation of survivin
expression, reporter constructs containing mutated putative YY1 binding
sites in the proximal survivin promoter were used in reporter assays.
Summary of Results: When YY1 was overexpressed, luciferase expression
was repressed 5Y10 fold. Further studies showed that knockdown of YY1
releases the survivin promoter from the observed repression and leads to a 3Y5
fold increase in promoter activity above basal levels. Furthermore, a low to
moderate decrease in Survivin protein was observed in Tet removal experiments. Site-directed mutagenesis confirmed the involvement of proximal
survivin promoter sites in YY1 overexpression-induced reporter repression.
Mutation of these binding sites on the survivin promoter showed abrogation
of the observed repression of survivin promoter activity.
Conclusions: These data suggest that YY1 is a novel repressor of survivin
transcription. A better understanding of how YY1_s regulation of survivin
expression in vivo affects cell cycle and cell death may provide new directions in therapeutics designed to prevent survivin overexpression in cancer.
17
REGULATION BY ESTROGEN AND PROGESTERONE OF
INTERFERON ALPHA SIGNALING IN HUMAN LEUKOCYTES
Zheng J, Hughes G University of Washington School of Medicine, Seattle, WA.
Purpose of Study: 90% of systemic lupus erythematosus (SLE) patients are
female, and the incidence peaks during reproductive years when estrogen (Es)
and progesterone (Pg) are at their highest. Studies show that Es increases the
risk of SLE in both humans and animal models, while Pg is protective.
Interferon alpha (IFN->) is a central pathogenic cytokine in SLE and can
directly activate multiple immune cell types to favor autoimmunity. Though
recent studies suggest a link between Es and IFN-> signaling in immune
cells, the relationship is not well understood; and very little is known about
the effects of Pg. To investigate the relationship between female reproductive
hormones and lupus autoimmunity, we tested the hypothesis that, in human
leukocytes, Es enhances IFN-> signaling while Pg suppresses it.
Methods Used: Peripheral blood mononuclear cells (PBMCs) were isolated
from blood of healthy donors. Cells were cultured in media, ethanol vehicle,
or physiologic concentrations of A-estradiol (Es), Pg, or a combination
of both hormones, with or without IFN->. Total RNA was isolated, and
expression of IFN-> inducible genes (CXCL10, MX1, PKR, IFIT1, ISG20)
was measured by quantitative PCR and normalized to housekeeping gene
18sRNA.
Summary of Results: Consistent with our hypothesis, we observed that
Es enhanced IFN-> signaling in human leukocytes; we did not observe an
effect of Pg. Es treatment significantly increased IFN-> induced expression
of CXCL10 (p=0.0273, two-tailed paired T-test). A similar trend was
observed for MX1, PKR and IFIT1. Es alone did not increase expression of
any of these genes. In addition, we observed that IFN-> signaling response
to Es was bi-modal, because PBMCs from some donors consistently showed
Es-sensitivity while others did not.
Conclusions: Our research shows for the first time that Es can enhance IFN> induced gene expression in human leukocytes of healthy donors, possibly
by regulating IFN-> signaling pathways. This suggests a novel mechanism by
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
which Es increases a woman_s risk of developing SLE. Additional experiments will allow us to determine whether Es regulates all or a subset of IFN>-inducible genes and what factors determine Es sensitivity, such as Es
receptor expression. We plan to extend these findings to PBMCs of SLE
patients to ask whether abnormal sensitivity or resistance to Es or Pg may
contribute to disease.
18
INHIBITION OF HERPES SIMPLEX VIRUS (HSV)
GLYCOPROTEIN_S gB, gD, gH-gL INDUCED CELL FUSION
AND VIRAL SPREAD VIA HEPARAN MIMICS
Choudhary S1, Tiwari V1,2 1Western University of Health Sciences, College
of Osteopathic Medicine of Pacific, Pomona, CA and 2Western University of
Health Sciences, College of Optometry, Pomona, CA.
Purpose of Study: Multinucleated giant cells (polykaryocytes) resulting
from virus-induced cell fusion are a hallmark of herpes simplex virus-type-1
(HSV-1) infection. However, the role of HSV-1 3-O-sulfated heparan sulfate
(3-OS HS) receptor during cell fusion is poorly understood. The goal of thisc
study was to understand the role of heparan mimics in blocking the HSV- 1
infection.
Methods Used: HSV-1 glycoprotein induced quantitative reporter gene
(luciferase) based cell fusion assay was used. The counting of multinucleated
giant cells (polykaryocytes) was achieved by using fluorescent microscopy.
Effector Chinese hamster ovary (CHO-K1) cells expressing various combinations of HSV-1 glycoproteins were co-cultured with the 3-OST-3 expressing primary cultures of human corneal fibroblasts (CF), a natural target
cell-type for HSV-1 infection. Pre-treatment of effector cells with heparinase
enzyme (4U/ml) and heparan mimics (5.6 nM) were used to demonstrate the
role of 3-OS HS during cell fusion.
Summary of Results: We demonstrated that cell-to-cell fusion and polykaryocytes formation required expression of four essential HSV-1 glycoprotein (gB, gD, gH-gL) and exhibited a very strong dependence on the
expression of heparan sulfate on CF. Further enzymatic removal of HS from
CF surface by heparinase-I treatment severely impaired the fusion reaction.
Interestingly, the incubation of effector cells expressing HSV-1 glycoprotein_s with heparan sulfate mimics inhibited significantly both membrane
fusion and polykaryocyte formation.
Conclusions: Our results indicate that 3-OS HS could play a crucial role in
HSV-1 induced cell-to-cell fusion during corneal eye infection. Generation of
specific inhibitors targeting against HSV-1 gD epitope that interacts with 3OS HS would be beneficial in the development of therapeutics to prevent viral
spread in the corneal stroma. Here, we provide evidence for the physiological
significance of 3-OS HS mediated cell-to-cell fusion.
19
ISLET MATRIX METALLOPROTEINASE-9 IS
DOWNREGULATED IN A MOUSE MODEL OF TYPE 2
DIABETES
Svy D1,2, Aston-Mourney K2,1, Kahn S2,1 1University of Washington,
Seattle, WA and 2VAPSHCS, Seattle, WA.
Purpose of Study: A pathological hallmark of the pancreatic islet in type
2 diabetes is aggregation of islet amyloid polypeptide (IAPP) into extracellular amyloid deposits, the process of which is toxic to A cells. We have
previously shown that (a) glucose dose-dependently increases amyloid
deposition in human IAPP (hIAPP) transgenic (T) mouse islets, (b) matrix
metalloproteinase (MMP)-9 degrades hIAPP, and (c) inhibition of MMP-9
activity increases amyloid deposition in hIAPP T islets. Whether high glucose regulates MMP-9 or tissue inhibitor of metalloproteinase (TIMP)-1, a
natural inhibitor of MMP-9, is unknown. Thus, we determined if downregulation of MMP-9 occurs in islets in vitro when cultured in high glucose
and/or in vivo in a diabetic mouse model.
Methods Used: Amyloid-forming hIAPP T and non-amyloid forming nontransgenic (NT) mouse islets were cultured for 7 days in 11.1, 16.7, or
33.3 mM glucose (n=4-6). Islets were also isolated from diabetic db/db and
control db/+ mice (n=8). To confirm the presence of amyloid in T islets after
culture, islet sections were stained with thioflavin S. MMP-9 and TIMP-1
mRNA levels were determined by real-time PCR using 18S as the endogenous control. MMP-9 activity was measured in islet conditioned medium by
gelatin zymography (n=3).
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Summary of Results: In cultured T and NT islets, neither MMP-9 nor
TIMP-1 mRNA changed with increasing glucose. In contrast, MMP-9
activity tended to increase at 33.3 mM glucose (NT at 11.1, 16.7, and
33.3 mM glucose: 1, 0.98T0.21, 3.61T0.95; T at 11.1, 16.7, and 33.3 mM
glucose: 1.22T0.06, 1.03T0.18, 3.92T1.60; p=0.06), but was not affected
by amyloid formation which was observed at 16.7 and 33.3 mM glucose. In
db/db mouse islets, MMP-9 mRNA decreased by 41% when compared to
db/+ islets (p=0.04).
Conclusions: Glucose did not downregulate MMP-9 activity in cultured
islets and therefore, decreased MMP-9 is not the mechanism by which glucose induces amyloid formation in this model. However, in a whole animal
model, MMP-9 mRNA was significantly decreased in diabetes. This suggests
that downregulation of MMP-9 by another component of the diabetic milieu,
such as elevated free fatty acids, may contribute to the increased amyloid
deposition associated with type 2 diabetes.
20
COLLAGEN SUBSTRATE SPECIFICITY OF PROLYL
3-HYDROXYLASES
Farnand AW1,2, Weis ME2, Kim LS2, Fernandes RJ2, Eyre DR2 1University
of Washington, Seattle, WA and 2University of Washington, Seattle, WA.
Purpose of Study: Current attention has focused on prolyl 3-hydroxylases
(P3Hs), a class of enzymes responsible for 3-hydroxylation of certain prolyl
residues in the collagen triple helix, forming 3-hydroxyproline (3Hyp).
Mutations in the genes encoding P3H1 and associated proteins of its enzyme
complex were found to cause recessive forms of severe osteogenesis
imperfecta (OI). The goal of this study was to investigate the expression
levels of the three P3H enzymesVP3H1, P3H2 and P3H3Vin a cell line
known to form 3Hyp at several sites in type II collagen, to better understand
their different substrate specificities.
Methods Used: Using qPCR, expression of the three P3H enzyme genes
was assayed in the rat chondrosarcoma cell line, RCS-LTC, and in normal
adult rat cartilage. Similarly, the expression profiles of these genes, and other
genes related to the functionality of the P3Hs, were compared qualitatively by
RT-PCR and were compared to expression levels in the human osteosarcoma
cell line, SAOS-2. Collagen produced by the RCS-LTC cells was analyzed for
3Hyp formation at known sites by mass-spectometry.
Summary of Results: The RCS-LTC cells produced mRNA for all three
enzymes, most prominently for P3H2, in contrast with normal adult rat
cartilage and SAOS2 cells which lacked mRNA expression for P3H2 and
P3H3 respectively. Quantitative comparison of P3H mRNA expression
between the RCS-LTC cells and rat cartilage yielded similar expression of
P3H1, a È2-fold increase in P3H2 and a È0.5-fold increase in P3H3
expression in the RCS-LTC cells. Mass-spectometric analysis of the RCSLTC collagen showed near complete 3Hyp formation at Pro944, a secondary
site previously found occupied in the type II collagen of vitreous, but not
cartilage.
Conclusions: The high relative abundance of P3H2 mRNA expression
coupled with high occupancy of Pro944 in the RCS-LTC matrix implicates
P3H2 in the formation of 3Hyp at Pro944. Taken together with previous
findings, it is possible that 3Hyp formation at Pro944 may be a signaling
mechanism for the prevention of cleavage of the collagen N-propeptide,
thereby maintaining the long, thin fibrils as seen in RCS-LTC cells and vitreous.
Funded under NCRR Grant TL1 RR 025016 and NIAMS Grant AR 036794
21
THIRDHAND SMOKE ADVERSELY AFFECTS FETAL LUNG
GROWTH AND DIFFERENTIATION
Chien K, Sakurai R, Torday J, Rehan V Los Angeles Biomedical Institute,
Torrance, CA.
Purpose of Study: Though nicotine is the main cigarette smoke constituent
linked to altered lung structure and function in in utero smoke exposed
infants, other constituents have not been thoroughly evaluated. This is particularly true for thirdhand smoke (THS), defined as tobacco smoke that sorbs
onto indoor surfaces and dust, where semi- and non-volatile chemicals and
particulates can undergo modifications to produce new toxicants that remain
on the surfaces, or later desorb and re-appear in the micro-environment. We
tested the hypothesis that THS exposure alters the growth and differentiation
of the developing lung by affecting the alveolar PPARF signaling known to
be essential for normal lung development.
93
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Methods Used: Embyronic day 19 fetal rat lung explants were exposed
to diluent, nicotine (110-8M or 110-5M), or 4-(methylnitrosamino)-1-(3pyridyl)-1-butanone (NNK) (110-8M or 110-5M), the main tobaccospecific N-nitrosamine constituent of THS, for 24h. Cell proliferation (BrDU
incorporation), cytotoxicity (LDH assay), and expression of key markers of
alveolar differentiation (surfactant protein B and C and PPARF expression,
choline incorporation into saturated phosphatidylcholine and triolein uptake)
were determined.
Summary of Results: Cell proliferation decreased and cytotoxicity increased significantly with NNK compared to nicotine (pG0.05), suggesting
more adverse effects of NNK. With NNK, choline incorporation into saturated phosphatidylcholine decreased significantly (pG0.05), while it increased
significantly with nicotine. Triolein uptake increased significantly with both
nicotine and NNK (pG0.05), though more robustly with NNK (pG0.05 vs.
nicotine). Western analysis showed significantly decreased PPARF and
increased fibronectin levels (pG0.05) with both NNK and nicotine exposure.
Conclusions: NNK affects lung growth and differentiation more robustly
than nicotine, possibly contributing significantly to in utero smoke exposureinduced pulmonary damage. These data can be used in designing specific
preventive and/or therapeutic strategies against the deleterious effects of in
utero THS exposure on lung development [Supported partially by grants from
the TRDRP (15IT-0250, 17RT-0170).
22
FACTORS CONTRIBUTING TO THE EFFICACY OF
THYMOGLOBULIN AS A TREATMENT FOR ACUTE
KIDNEY GRAFT REJECTION
Liu Y1, Kuo H1, Jiang Y2, Bunnapradist S1 1UCLA Medical School, Los
Angeles, CA and 2UCLA, Los Angeles, CA.
Purpose of Study: To examine the outcomes of Thymoglobulin treatment
for acute rejection and to identify risk factors associated with graft failure
using the UCLA Kidney Transplant Database.
Methods Used: Between February 2000 and June 2010, there were 517
hospital admissions for possible kidney allograft rejection. Among these,
there were 114 unique patients treated with Thymoglobulin for acute rejection, and 98 were included in the analysis, with a median follow up time of
567 days (16 patients did not have sufficient follow-up data). An analytical
file was created by retrospective chart-review and baseline characteristics
were collected. The Kaplan-Meier product limit method was used to estimate
the survival rate. Univariate and multivariate survival analyses were performed using the Cox proportional hazard model.
Summary of Results: Graft failure occurred in 36 of the 98 patients
(36.7%) with a median time between Thymoglobulin treatment and graft
failure of 567 days (25Y75%: 152Y1265 days). In multivariate analysis, factors
associated with increased risk of graft failure included doses of Thymoglobulin received (Q9) and number of biopsies performed on the current allograft
(Q1). Factors associated with decreased risk of graft failure included creatinine level at time of Thymoglobulin treatment (G3.0 mg/dL) and identification of vascular rejection versus non-vascular rejection on the biopsy
performed immediately prior to Thymoglobulin treatment.
Conclusions: Resolution of acute rejection is vital to allograft survival and
Thymoglobulin is a last-line treatment for steroid-resistant acute rejection.
Our results indicate that the doses of Thymoglobulin received, the creatinine
level at time of Thymoglobulin treatment, the number of prior biopsies, and
the presence of non-vascular rejection are associated with increased risk
for graft failure. Individual chart-review permitted access to biopsy data not
available in database studies, and the analysis suggests that perhaps Thymoglobulin treatment is less efficacious in acute rejection with chronic
changes. In patients with risk factors found to be significant for graft failure,
nephrologists may choose to lower or omit the use of Thymoglobulin as its
benefits may be outweighed by its recognized adverse effects in the long run.
23
ASTROCYTE MATURATION AND REACTIVITY IN
PEDIATRIC EPILEPSY SYNDROMES
Kwon EE1,2, Mathern G3, Wanner I2 1Western University of Health Sciences,
Pomona, CA; 2University of California, Los Angeles, Los Angeles, CA and
3
University of California, Los Angeles, Los Angeles, CA.
Purpose of Study: Astrocytes are directly involved in the blood brain
barrier and influence potassium buffering through aquaporin4 (AQP4) water
94
&
Volume 59, Number 1, January 2011
channels. AQP4 is known to buffer metabolites during neural injury and
epileptogenesis. GFAP (glial fibrillary acidic protein), a prototypical marker
of astrocytes, is upregulated and reveal astrocytes undergo structural changes
such as hypertrophy and loss of domain organization. Severe forms of
astrogliosis lead to the formation of glial scars and scar-like penumbras
around necrotic or ischemic areas. To further characterize astrocytes in
human epilepsy, we analyzed hallmark markers of astrocytes in pediatric
surgical biopsy specimens of the neocortex.
Methods Used: Cortical tissue were resected from pediatric patients with
epilepsy disorders including Cortical Dysplasia, Rasmussen_s Encephalitis,
and epilepsy due to ischemic stroke primarily from central operculum and
parieto-temporal lobe regions. Specimens were vibratome-sectioned and
stained for GFAP and AQP4 using flourescence immunohistochemistry
and images were binarized and quantified for astrocyte abundance and
coverage as a measure of reactivity.
Summary of Results: Our results show that abundance as well as network
complexity varied in each zone. One striking finding was a noticeable
sparseness, even lack GFAP fluorescence in the grey matter of infant cortical
biopsies. This finding, together with the presence of immature radial glia
marker expressing cells in that zone (BLBP-brain lipid binding protein)
suggests that GFAP-positive astrocytes did not significantly populate the grey
matter until around 4 years of age. This was found to be the case in genetic as
well as acquired epilepsy etiologies, irrespective of case severity, which
suggests a developmental feature of protoplasmic astrocytes rather than
pathological.
Conclusions: These tools allow us to determine reproducible quantitative
differences in astrocyte abundance, morphology and reactivity that may help
define gliosis severity. These findings would help us understand the role of
astrocytes in epilepsy and also be useful in histopathological diagnosis of
various epilepsy syndromes.
24
GENETIC POLYMORPHISMS OF CYTOCHROMES P450
AND PEDIATRIC ASTHMA CONTROL BY INHALED
CORTICOSTEROIDS
Stockmann C1, Fassl B1, Nkoy F1, Stone B1, Willis L1, Gaedigk R2,
Leeder S2, Ward R1 1University of Utah, Salt Lake City, UT and
2
University of Kansas Children’s Hospital, Kansas City, KS.
Purpose of Study: Inhaled corticosteroids (ICS) are mainline treatments for
persistent asthma in children. Up to 10% of children with persistent asthma
fail ICS therapy for unclear reasons. Genetic polymorphisms of cytochromes
P4503A (CYP3A) have been shown to be associated with altered metabolism
of ICS which might have an effect on their therapeutic effectiveness. Fluticasone, a widely used ICS, is metabolized primarily by CYP3A4 whose
metabolism may be decreased by a polymorphism in intron 6. Objectives of
this study are to 1) determine the frequency of genetic variants in CYP3A4
(intron 6 SNP rs35599367,C9T), in children treated with fluticasone for
difficult to control asthma and 2) to describe and compare CYP450 genotypes
in these children with and without previous hospital admissions for asthma.
Methods Used: We enrolled children aged 2Y17 years with a diagnosis of
asthma from Primary Children_s Medical Center. We collected saliva samples
and analyzed for CYP450 genetic polymorphisms at the Developmental
Pharmacology and Experimental Therapeutics Laboratory at Kansas City.
Single nucleotide polymorphisms for 9 alleles that increase or decrease
protein expression and CYP3A activity were determined along with asthma
severity, preventive medication use, and number of hospitalizations for acute
asthma within the preceding 12 months.
Summary of Results: Of 96 children enrolled, 23 were treated with fluticasone; all of these were classified as not-well-controlled asthmatics by NIH
guidelines. Of these 23 children, 14 patients were admitted for asthma in the
previous 12 months, 14/14 featured an rs35599367 C/C genotype consistent
with greater CYP3A4 mRNA level and enzyme activity. Of the 9 patients not
requiring hospitalization within the last 12 months, 5 (55.6%) had at least one
variant rs35599367 allele, consistent with slower metabolism of fluticasone.
There was no difference in the distribution of other CYP3A4, CYP3A5,
and CYP3A7 polymorphisms.
Conclusions: These preliminary findings are consistent with the hypothesis
that slower metabolism of ICS may improve their effectiveness for treatment
of asthma. Additional patient studies are needed to confirm or refute this
association.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
25
COMBINATORY EFFECT OF BMP-2 AND APATITE COATED
SCAFFOLDS ON THE OSTEOGENIC DIFFERENTIATION
OF HUMAN MESENCHYMAL STEM CELLS
Davis HE, Case E, Miller S, Leach JK University of California, Davis,
Davis, CA.
Purpose of Study: It is widely accepted that osteoconductive materials
promote osteogenesis in vivo. However, the impact of 3D osteoconductive
substrates coupled with soluble signals on progenitor cell differentiation is
not well known. In this study, we investigated the influence of carbonate
apatite on the osteogenic differentiation of human mesenchymal stem cells
(hMSCs) seeded in biodegradable poly(lactide-co-glycolide) (PLG) scaffolds
in conjunction with the osteoinductive growth factor BMP-2.
Methods Used: Apatite coated PLG scaffolds were formed by incubating
hydrolyzed microparticles in modified simulated body fluid (mSBF) prior to
being combined with NaCl and compressed at 1500 psi for 1 min. Samples
were then exposed to high pressure CO2 (800 psi) for 16 h. NaCl was
removed by leaching the matrices in distilled H2O for 24 h. We cultured
hMSCs on the scaffolds in osteogenic media supplemented with 25, 100 or
200 ng/ml of BMP-2. The in vitro osteogenic differentiation of hMSCs was
measured by quantifying ALP activity, DNA content, and cell secreted calcium after 3, 7, 14, 21 and 28 d in osteogenic conditions. qPCR was performed for Runx2, SP7, Col1a1, lbsp, and Sparc at 7 and 21 d.
Summary of Results: No significant differences between nonmineralized
and mineralized scaffolds were detected in ALP activity at 0 and 200 ng/mL
concentrations, but mineralized scaffolds had increased levels after 7 d at 25
and 100 ng/mL. Mineralized scaffolds had increased levels of cell-secreted
calcium at all time points. Increasing BMP-2 concentration resulted in a rise
in both ALP and calcium levels. There was no difference between Runx2 and
SP7 levels in mineralized and nonmineralized substrates, but an increase in
SP7 was observed with the addition of BMP-2. At 21 d mineralized substrates
had increased levels of lbsp and decreased levels of Sparc compared to
nonmineralized substrates.
Conclusions: The present data indicate that a combination of apatite and
BMP-2 do not simply enhance the osteogenic response of hMSCs, but act
through different and possibly opposing pathways. Thus multiple signaling
strategies may be necessary to achieve optimal bone regeneration.
Behavior and Development
Concurrent Session
8:30 AM
Friday, January 28, 2011
26
EXAMINATION OF VERBAL INTELLIGENCE AND
PHONOLOGICAL PROCESSING IN CHILDREN WITH
AUTISM SPECTRUM DISORDERS AND THEIR
FAMILY MEMBERS
Thomson AL1, Webb SJ2, Bernier R2, Wijsman E2 1University of
Washington School of Medicine, Seattle, WA and 2 University of
Washington, Seattle, WA.
Purpose of Study: Autism Spectrum Disorders (ASD) have a strong
genetic influence, and a broader autism phenotype (BAP) has been observed
in first-degree family members. It is well known that children with autism
have an array of communication deficits, including phonology, semantics,
syntax, and pragmatics. The overall aim of this project was to better understand verbal language use in individuals with ASD and their family members.
Methods Used: A total of 279 family members from families with 2 or more
ASD children participated. Verbal intelligence quotients (VIQ), a measure of
vocabulary and reasoning, were assessed with age-appropriate Wechsler
scales. Non-word repetition (NWR), a measure of phonological processing,
was assessed with the CTOPP.
Summary of Results: As expected, ASD children exhibited lower VIQ
compared to mothers, fathers, and unaffected siblings (F(3,274) = 59.8,
p G.01).To examine decreased VIQ in the context of overall cognitive function, discrepancy scores were calculated between verbal and non-verbal IQ.
ASD children_s IQ discrepancy scores were significantly greater than relatives (F(3,274) = 19.8, p G.01). Both our higher-functioning (HF) and lower-
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
functioning (LF) ASD groups exhibited a mean discrepancy pattern that
suggested better non-verbal IQ compared to VIQ (pG.05). For phonological
processing, mean performance for all groups was significantly below the
norm on the NWR task, however siblings performed relatively better than
ASD children and fathers (F(3,256) = 2.3, pG.05). Investigation of the relationship between VIQ and NWR revealed significantly higher ratio scores in
ASD children and siblings compared to parents (F(3,253)=11.976, p G.01).
Conclusions: These results suggest that relatively poorer phonological
processing may be an endophenotype of ASD and a part of the BAP. The ratio
of the phonological processing to VIQ suggests that the relation between
these two domains changes with age, but not affected status. Taken together,
these data suggest that multiple aspects of language processing contribute to
both autism and the broader phenotype and will be important quantitative
traits for genetic analyses.
27
MISSED OPPORTUNITIES IN HIGH RISK INFANT
FOLLOW-UP?
Tang B1, Huffman L1, Feldman H1, Gray E1, Kagawa K2, Gould JB1
1
Stanford University School of Medicine, Palo Alto, CA and 2State of
California, Sacramento, CA.
Purpose of Study: To evaluate rates of referrals to state-funded early
intervention (EI) programs for high risk infants demonstrating significant
developmental delay during the 1st year of life following NICU discharge. In
California, high risk infant follow-up (HRIF) programs are supported by
California Children_s Services (CCS). Infants discharged from the NICU
qualify for HRIF based on CCS-defined neonatal medical risk factors. A
central goal of HRIF is to identify children with developmental delays and
refer them to EI that provides appropriate therapeutic services.
Methods Used: A primary analysis of a population-based data set from a
California network of HRIF programs. Standardized developmental assessments were conducted at the 1st HRIF visit at 4Y8 months of age, adjusted for
prematurity. Scores were reported as a developmental quotient (DQ) based on
normative data for each individual test. For this study, infants were included
in the analyses if they had a DQ e70 (Q2 standard deviations below the mean)
in Q1 subtest domain (e.g. motor). Of the included infants, we compared the
proportion who were and were not referred to EI.
Summary of Results: Of the 1880 high risk infants who had a standardized
developmental assessment at the 1st HRIF visit, 342 (18%) had Q1 domain
DQs in the 2Y3% range of the general population. Two-thirds (n=235) had no
EI before the 1st visit. Of those infants, less than half were referred to EI
(Table). This finding persisted across birth weight group classifications and
for infants who had Q2 domain DQs e70.
Conclusions: An alarmingly high proportion of high risk infants are not
referred to EI even after they are identified, using standardized testing, as
having a developmental delay in Q1 domain. Factors associated with the nonreferral group will be explored. Subsequent HRIF visits will determine
whether a delay in referral affects long term developmental outcomes. These
data have utility for the quality improvement of HRIF programs across
the state.
EI Status of Infants with DQs e70 at the 1st HRIF Visit
28
USE OF DIFFUSION TENSOR IMAGING IN THE
EVALUATION OF BRAIN INJURY IN ADOLESCENTS
BORN PRETERM
Gaman-Bean S1, Yeatman J1, Lee E1, Yeom K2, Feldman H1 1Stanford
University, Palo Alto, CA and 2Stanford University, Palo Alto, CA.
Purpose of Study: Subtle, diffuse, non-cystic white matter injuries have
been found in pretern infants and are often missed by standard MRI. Diffusion tensor imaging (DTI) is a technique capable of examining the structure
and organization of white matter at the microscopic level and is particularly
95
Western Regional Meeting Abstracts
Journal of Investigative Medicine
useful for detecting this type of injury. The hypothesis is that the preterm
group will have indications of injury or immaturity in the corticospinal and
somatosensory tracts when comparing the preterm and full-term groups.
Methods Used: DTIs were collected and processed for 50 subjects, both
born preterm and full-term, between 9-16 years of age. Using tractography,
the corticospinal and somatosensory tracts were tracked from the cerebral
peduncle to the primary motor and sensory cortices in each individual. We
computed the diffusion parameters, axial and radial diffusivity, and the
fractional anisotropy along the trajectory of the tracts. These were then
averaged for both groups and compared.
Summary of Results: The axial diffusivity of the preterm group is higher
along the corticospinal and somatosensory tracts when compared to the fullterm group. The radial diffusivity is similar between the two groups along the
tracts. The fractional anisotropy is similar between the two groups, but is
higher in the preterm group at several points along the tracts.
Conclusions: Analysis suggests that there are detectable differences
between the corticospinal and somatosensory tracts of the two groups and that
perinatal injury may show dectectable differences into adolescence. Prior
studies have focused on fractional anisotropy, but this study shows that there
are detectable differences in the axial diffusivity between the two groups.
Future analyses will include a more detailed examination of the preterm
group to look for outliers and to compare the findings of the preterm group
with their motor exam.
An example of the findings: left corticospinal tract.
29
EXAMINATION OF BASELINE ATTENTION DEFICIT/
HYPERACTIVITY DISORDER(ADHD) COMORBIDITIES
AND THEIR ASSOCIATION WITH CAREGIVER STRESS
AND DEPRESSION
Pascoe PE1, Rockhill C1, Myers K1, Palmer N2, Vander Stoep A3, Geyer J2,
McCarty C1 1University of Washington, Seattle, WA; 2Seattle Children’s
Hosptial, Seattle, WA and 3University of Washington, Seattle, WA.
Purpose of Study: ADHD comorbidities have been associated with differential treatment outcomes and higher levels of caregiver stress and
depression. This study examines: 1) the prevalence of ADHD comorbidites
identified using the Diagnostic Interview Schedule for Children (DISC)
versus the Child Behavior Check List (CBCL), and their agreement in the
diagnosis of ADHD comorbidities; 2) the association between ADHD with
and without comorbidities (anxiety disorders, oppositional defiant disorder
(ODD) or both) and caregiver stress and depression.
Methods Used: This study used baseline data from the Children_s ADHD
Telemental Health Treatment Study (CATTS), a randomized clinical trial
examining the efficacy of telepsychiatry for the management of ADHD
among youth referred by their primary care providers. Comorbidity was
assessed using the DISC and the CBCL. Caregiver stress and depression were
measured with a battery of instruments.
Summary of Results: The CBCL and DISC identified a similar number of
youth for the diagnosis of ADHD without comorbid disorders (36% for both
CBCL and DISC) and ADHD with both comorbid oppositional defiant disorder (ODD) and anxiety disorder (16% for C-DISC, 19% for CBCL). The
CBCL was more likely to identify youth with an anxiety disorder while the
DISC was more likely to identify youth with ODD. Agreement between
CBCL and DISC was poor, with kappas ranging from 0.13 to 0.28. Greater
caregiver stress and depression was associated with the presence of comorbidities with ADHD, especially when comorbid diagnoses were based upon
the CBCL and when youth had both ODD and anxiety diagnoses.
96
&
Volume 59, Number 1, January 2011
Conclusions: The results of this study highlight differences between wellknown and frequently used measures regarding identification of ADHD
comorbidities. In addition, this study underlines the need to recognize caregiver stress and depression among youth with ADHD and comorbid
diagnoses.
30
LOW GLYCEMIC LOAD EXPERIMENTAL DIET MORE
SATIATING THAN HIGH GLYCEMIC LOAD DIET
Chang K1, Schwarz Y2, Lampe J1,2, Neuhouser M2 1University of
Washington School of Medicine, Seattle, WA and 2Fred Hutchinson
Cancer Research Center, Seattle, WA.
Purpose of Study: Clinicians need effective yet simple strategies to offer
overweight and obese patients in need of weight reduction, especially with
evidence suggesting metabolic disorders are a risk factor for developing and
dying from cancer. Of interest in this regard is whether particular food types
increase satiation, which may reduce overall food intake. With a randomized
controlled feeding study, we investigated the effect of low vs. high glycemic
load (GL) experimental diets on satiety. We also examined whether BMI,
gender, and leptin concentration influenced results.
Methods Used: 81 healthy adult participants, half of whom were overweight or obese, completed two 4 week feeding periods (low GL and high
GL) which were separated by a 4 week washout. Diets were matched in
calories and macronutrients (except for fiber) and differed only in GL. All
meals were prepared in a controlled manner in a metabolic kitchen. Participants completed a visual analog satiety survey after each feeding period and
T-tests were used to compare mean scores. Serum leptin was assayed from
blood draws taken after each feeding period.
Summary of Results: Participants reported feeling significantly more
satiated on the low GL diet than on the high GL diet (p=0.047). Participants
also reported having more food cravings on the high GL diet than on the low
GL diet (pG0.001). In subgroup analysis, women demonstrated significantly
less hunger and more satiety on the low GL diet (p=0.05, pG0.01). There was
no difference in hunger or satiety for men between the two diets. Participants
of normal body fat percentage (G25% for men; G32% for women) and normal
BMI (G 25.0) reported the food being tastier on the low GL diet than on the
high GL diet (p=0.04, p=0.05). Leptin concentration did not significantly
vary amongst participants between the two diets.
Conclusions: Our data demonstrate that a low GL diet is more satiating than
a high GL diet, especially for women. Reducing GL may be an effective way
to lower the amount of food consumed and improve compliance with weight
reduction prescriptions. A low GL diet may thus be a potential tool to reduce
the risk of developing and dying from cancer by lowering rates of overweight
and obesity.
31
GESTATIONAL AGE ASSOCIATED WITH DEVELOPMENTAL
SCORES IN HEALTHY CHILEAN INFANTS BORN
BETWEEN 37 AND 42 WEEKS GESTATION
Dolgonos O1, Martinez S1, Blanco E1, Lozoff B2, Gahagan S1,2 1University
of California, San Diego, La Jolla, CA and 2University of Michigan, Ann
Arbor, MI.
Purpose of Study: To test the hypothesis that gestational age (GA) is
associated with mental and psychomotor development scores in healthy
infants born between 37 and 42 weeks.
Methods Used: The sample included 1739 participants of a longitudinal
study, which began as an iron-deficiency anemia preventive trial in Santiago,
Chile. Infants were selected for the trial if they weighed 3 kg or more at birth,
were born between 37Y42 weeks of gestation, and were healthy. Children
born between 37Y39 weeks of gestation were compared with children born
at 40Y42 weeks of gestation. The Bayley Scales of Infant Development
at 12 months were used to assess development. Using generalized linear
modeling, we assessed the association between gestational age and developmental status controlling for birth weight, sex, family socioeconomic
status, and Home Observation for Measurement of the Environment (HOME)
Inventory.
Summary of Results: Of the 1739 infants, 851 were born at 37Y39 weeks
GA and 888 were born at 40Y42 weeks GA. Infants born at 37Y39 weeks had
mean Bayley Mental and Psychomotor Development scores that were 2.0 and
2.9 points lower than children born at 40Y42 weeks of gestation, respectively
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
(pG.05). Higher gestational age was associated with higher Bayley Mental
(B=1.6, pG.05) and Psychomotor (B=2.3, pG.05) Development scores after
controlling for covariates. All covariates were statistically significant (pG.05)
except birth weight for the mental development score and sex for the psychomotor score. Assignment to iron or usual nutrition, and occurrence of iron
deficiency anemia or not, were not significant in the models.
Conclusions: In a cohort of healthy full term infants from Santiago, Chile,
developmental status at 12-months was significantly lower in those born at
37Y39 weeks GA than those born at 40Y42 weeks GA. Further research could
assess whether these differences persists beyond infancy.
32
PSYCHOLOGICAL COUNSELING FOR PARENTS
CARRYING A FETUS DIAGNOSED WITH A CONGENITAL
HEART DISEASE
Creel B1, Evans WN1,2, Acherman RJ1,2, Rollins RC1,2, Castillo WJ1,2,
Luna CF1,2, Restrepo H1,2 1Children’s Heart Center Nevada, Las Vegas, NV
and 2University of Nevada, School of Medicine, Las Vegas, NV.
Purpose of Study: To assess the emotional state, changes in daily life, and
emotional support systems in newly diagnosed parents of a fetus with a
confirmed diagnosis of structural heart disease.
Methods Used: In December 2009, we began a new program of psychological counseling for couples carrying a fetus with heart disease. This report
includes the first 16 couples in this program. The couples responded to a
questionnaire developed by the investigators focusing on the emotional state,
available support systems, and the effects of prenatal knowledge of the
diagnosis.
Summary of Results: Mean mothers_ age was 29.1 T 6.7 years (range:
15Y36 years). The group of couples was composed of 7 Caucasians, 2 Hispanics,
2 Asians, and 5 from other races.
Table 1 summarizes the relative percentage of emotional states reported
by couples after the diagnosis of carrying a fetus with a congenital heart
disease.
Knowing the diagnosis in advance was useful in 15 out of 16 couples in
helping them to prepare, economically, mentally, and emotionally for the
birth of the child, only 1 couple responded being too overwhelmed and unsure
of their feelings around this question.
Daily life was impacted with negative emotions in 15 out of 16 couples;
one couple did not answer the question.
13 out of 16 couples reported that the main support systems in helping
cope with the diagnosis were their family and religious groups; the remaining
3 couples reported friends and others as their main support.
Conclusions: We found that in the vast majority of parents carrying a fetus
with a diagnosis of a CHD, negative emotional states impact their daily life
and family and religious groups are the main support systems to cope with the
diagnosis. These results showed that couples with a fetus with a CHD are in
need of psychological counseling in helping cope with the diagnosis and in
preparing for a life with a medically challenged child.
TABLE 1.
33
COGNITIVE PROFILES IN AUTISM SPECTRUM DISORDERS
Vincent L2, Bernier R1 1University of Washington School of Medicine,
Seattle, WA and 2University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Given the phenotypic and genetic heterogeneity of
Autism Spectrum Disorder (ASD), identification of accurate and easily
identifiable subtypes could improve etiologic understanding, intervention
strategies and prognostic recommendations. Using 1280 individuals with
ASD ascertained through the Simons Simplex Collection, our purposes were
to investigate the existence of domain discrepancy profiles based on verbal
and nonverbal intelligence scores, and to explore the relationships between
cognitive profiles and other measurable outcomes.
Methods Used: Based on stringent requirements for identifying significant
differences in verbal and nonverbal IQ scores, a chi-squared test identified
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
subjects as having either a nonverbal 9 verbal intelligence split, no split
(verbal = nonverbal intelligence), or a nonverbal G verbal intelligence split.
Chi-squared analysis was used to look at gender breakdown within profile
groups. Relationships between these profiles and other aspects of autism
symptomatology, including socialization skills, fine motor skills, adaptive
behaviors, head circumference, and autism severity were analyzed using
multivariate analyses of variance. Age was included as a covariate.
Summary of Results: A significantly greater number of individuals with a
cognitive split was found in the ASD group than would be expected. While no
relationships between cognitive profile and adaptive behavior or fine motor
skills were identified, results indicate that individuals with a nonverbal 9
verbal profile showed greater overall autism symptom severity according to
clinician observation and individuals with a nonverbal G verbal discrepancy
profile showed greater social impairment according to parent report.
Conclusions: These findings indicate that there is an increased rate of
nonverbal and verbal intelligence discrepancies in ASD and suggest that
cognitive profiles within the autism spectrum may provide a useful method of
identifying ASD subtypes. Further investigation of these cognitive profile
subtypes may elucidate differential genetic mechanisms and provide guidelines for determining the maximally effective interventions tailored to ability
profiles to further improve outcomes.
34
ARE THERE CHILDREN IN NEED OF EARLY
INTERVENTION SERVICES THAT ARE BEING MISSED
BECAUSE THEY DO NOT HAVE A DIAGNOSIS OF A HIGH
RISK CONDITION?
Weigang T1, Talmi A2 1University of Colorado Denver, Aurora, CO and
2
University of Colorado Denver, Aurora, CO.
Purpose of Study: Early Intervention services have been shown to be
effective resources for young children (under 3 years old) who, either are
developmentally delayed (DD) or have a condition that puts them at a high
risk of becoming DD. These services are most effective when utilized as early
as possible. A little over 2% of the pediatric population under 3 years old is
enrolled in Early Intervention services, but research indicates that as many as
13% of the children under 3 years old may be eligible for these services
(Rosenberg et. al 2008). The current study aimed to look at patterns of
pediatric provider referrals in the presence of an abnormal score on a parent
report child developmental screen (Ages and Stages Questionnaire (ASQ)).
Specifically, the study was designed to confirm findings reported by
Silverstein, et al (2006), who reported that the majority of pediatricians they
surveyed thought that an established diagnosis of a high risk condition was
important when deciding whether to refer a child to Early Intervention. Our
study aimed to answer the following questions: 1) is there a population of
children without established high risk conditions, but who show signs of DD
and are not being referred to Early Intervention services? 2) Do consultations
with mental health liaisons, who are collocated within the primary care
clinic(CLIMB), have any effect on these referral rates?
Methods Used: A retrospective study was done by extracting data from
electronic medical records of all well child checks (WCCs), of children under
3, done at an outpatient Child Health Clinic in a Children_s Hospital between
March of 2009 and April of 2010. Data was obtained by a computer query of
all WCCs documented in the electronic medical records. A computer query
and manual review of these records were used to identify all WCCs that had
an ASQ completed during the visit. The records with abnormal ASQ scores
were analyzed.
Summary of Results: Data analysis is still pending but will include: the
percent of ASQs with abnormal scores, Early Intervention referrals generated, the number of children with established high risk conditions, and
whether CLIMB consults had a significant impact on referrals.
Conclusions: Pending
35
THEORY OF MIND IN CHILDREN AT RISK FOR
DEVELOPING PSYCHOSIS
Reidy RE, Hunter SK University of Colorado Denver School of Medicine,
Denver, CO.
Purpose of Study: Theory of Mind (ToM), the ability to appropriately
attribute mental states to the self and others, is a cognitive ability that normally develops between three and five years of age (Wellman & Liu, 2004).
97
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Theoretically, psychosis may be related to deficiencies in both monitoring
and interpreting intentions of the self and others (Frith, 1992). A large
number of studies show that schizophrenic patients have poor performance
on ToM tasks (see Brune, 2005) and deficits in ToM performance have been
found in first-degree relatives of schizophrenic patients (Anselmetti et al.,
2009), suggesting that poor performance on ToM tasks may be an endophenotype for schizophrenia. Surprisingly, early impairments in ToM have
not been studied in the children of schizophrenics.
Methods Used: Children with at least one parent meeting DSM IV criteria
for diagnosis of schizophrenia or other psychotic disorder and children with
no family history of psychosis are given tasks at 40 and 48 months of age to
assess ToM development. The first is a locations false belief task which
requires the child to predict where a protagonist will search for an object
based on a false beliefs about the object location. The second task is a
contents false belief task and requires the child to attribute knowledge about
the contents of a mislabeled box to the self and others. The Dimensional
Change Card Sort is also being administered in order to analyze interactions
with executive function, another endophenotype for schizophrenia.
Summary of Results: Thirty children have completed the study to date and
preliminary data show children_s scores significantly increase with age, as
predicted. Of the thirty children who have completed the study, three have a
parent with a diagnosis of schizophrenia or other psychotic disorder, thus the
study currently lacks sufficient power to analyze differences in performance
between conditions of parental mental health status.
Conclusions: We hypothesize that poor performance on ToM tasks is an
endophenotype for schizophrenia and other psychotic illness that will be
detectable early in life. Longitudinal analysis is hypothesized to show a
significantly larger increase in task performance for children with no family
history of psychosis.
36
THE EXPERIENCES OF AUTISTIC YOUTH LIVING IN
RURAL COMMUNITIES
Herring JC 1University of Washington, Woodinville, WA and 2Seattle
Children’s Hospital, Seattle, WA.
Purpose of Study: As the American economy becomes more knowledgebased and the job market more competitive successful transition from adolescence to adulthood is even more critical, particularly for students with
disabilities like Autism. The NLTS2 is a ten-year transitional study that
provides researchers, educators and policy makers at the national level the
opportunity to examine the post-secondary experiences of youth with disabilities. The transitional and social experiences of youth with Autism is a
recent topic of research and requires much more investigation. The purpose
of this study is to provide a brief description of the transition, education, and
social experiences of Autistic youth living in rural communities.
Methods Used: The samples for this prospective study were taken from
Wave 1 and Wave 4 of data collection. Data was obtained from both samples
for youth that were 1) Diagnosed with Autism and 2) Attended a school
located in a rural community. Data was analyzed using the social sciences
statistical analysis software, SPSS (Statistical Package of the Social
Sciences, 2008).
Summary of Results: The majority of youth in this sample (ages 13Y16)
have trouble conversing, speaking clearly, understanding others, and using an
appropriate tone. Nearly half of parents (47%) stated that their child had never
gotten together with any friends in the past 12 months, while more than half
(54%) reported that their child had never been invited to any social activities
in the past 12 months. Additionally, 53% of respondents said that their autistic youth never talked to friends on the phone and nearly 66% reported that
their child never interacted with others using email or chatrooms.
Conclusions: Autism and the Associated Spectrum Disorders are a complex
interaction of impairments in verbal/non-verbal communication, behavior,
and social interaction. The results of this study suggest that Autistic youth
living in rural areas are mostly unengaged socially and in their community, as
well as unprepared for the transition to employment and post-secondary
education. More research is needed to further examine the disparities faced
by Autistic youth in rural communities, how to reform the transition process
for this population so as to achieve transition success, and analyze the
influence of youth experiences on future outcomes.
98
&
Volume 59, Number 1, January 2011
Cardiovascular I
Concurrent Session
8:30 AM
Friday, January 28, 2011
37
INDUCTION VS. NON-INDUCTION: THE SAGA CONTINUES
Kawano M, Goldstein Z, Hamilton M, Kobashigawa J Cedars-Sinai Heart
Institute, Beverly Hills, CA.
Purpose of Study: In cardiac transplantation, induction immunosuppression therapy has been administered in hopes of the recipient acquiring tolerance to the donor organ. There have been no randomized trials that have
demonstrated benefit to the use of induction therapy which usually includes
cytolytic drugs, such as antithymocyte globulin (ATG). 60% of the centers in
the United States do not use induction, but rather, triple drug immunosuppression with a regimen of tacrolimus, mycophenolate mofetil, and steroids.
The 40% that do use induction typically utilize ATG. This observational
study reviews two large, local programs in the same era and assesses the long
term outcome of heart transplant patients who were administered induction
versus no induction for heart transplantation.
Methods Used: We evaluated 1207 patients transplanted between 1994 and
2010, and divided them into those that received induction therapy and those
that did not. These patients were assessed for long term outcomes, including
5-year actuarial survival, freedom from cardiac allograft vasculopathy (CAV,
stenosis Q50%), freedom from non-fatal major adverse cardiac events
(NF-MACE, MI, CHF, PTCA, pacemaker, stroke, new peripheral vascular
disease), and 1-year freedom from any-treated rejection.
Summary of Results: The non-induction group (n=766) exhibited greater
freedom from NF-MACE compared to the induction group (n=441)(93% vs.
85%, pG0.001), mostly due to less patients developing CHF (5% vs. 9%,
p=0.009). The non-induction group also demonstrated a trend towards greater
5-year freedom from CAV compared to the induction group (87% vs. 83%,
p=0.07). However, there was no significant difference between groups in
terms of 5-year actuarial survival (78% vs. 78%, p=0.85). Interestingly, there
was also no difference in 1-year freedom from rejection in the induction
group, which used more aggressive early immunosuppression therapy.
Conclusions: Induction immunosuppression therapy does not appear to
have an advantage over non-induction immunosuppression. Specifically,
there was no difference in 1-year rejection and patients on induction therapy
demonstrated significantly higher incidence of NF-MACE as well as a trend
toward increased frequency of 5-year CAV. The cause of increased incidence
of NF-MACE in the induction group remains unknown. A randomized trial
is still needed to assess any benefit of induction therapy.
38
PRE-TRANSPLANT BLOOD TRANSFUSION CAUSING
SENSITIZATION IN PATIENTS AWAITING HEART
TRANSPLANTATION
Davidoff J, Kawano M, Goldstein Z, Hamilton M, Kobashigawa J CedarsSinai Heart Institute, Beverly Hills, CA.
Purpose of Study: Sensitization is the development of circulating antibodies in patients awaiting heart transplantation. Sensitization in these
patients can mean a longer waiting time in order to find a compatible donor
organ. One of the major causes of sensitization is the administration of blood
transfusions in the pre-transplant time period. However, it is not known what
the risk of blood transfusions is in terms of causing sensitization. We sought
to determine the risk of blood transfusions in patients awaiting heart
transplantation.
Methods Used: We reviewed 836 patients awaiting heart transplant between
1994 and 2008 and found 63 patients with baseline and subsequent blood
transfusions and panel reactive antibodies (PRAs) within one year prior to
transplantation. Sensitization was defined as any PRA screen greater than
baseline. Patients were divided into 3 groups depending on baseline PRA:
Group A = 0% (N=22), Group B = 0-10% (N=20), and Group C 9 10%
(N=21).
Summary of Results: 36% of Group A (baseline PRA=0%) patients
developed circulating antibodies after a mean of 1.4 + 0.5 blood transfusions.
Group B and Group C patients also were noted to have increases in circulating antibodies from baseline in 45% and 52% of patients, respectively.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
The risk of 1 transfusion vs. 6 transfusions did not appear to markedly
increase risk of sensitization.
Conclusions: The risk of sensitization following blood transfusions is significant although the extent of antibody production appears to be limited in
Group A-type patients (baseline PRA = 0%), as most of these patients do not
develop PRAs 9 10%. For patients with pre-existing circulating antibodies,
increase in PRA level is also significant. Withholding of blood transfusions to
prevent sensitization and further sensitization in patients with pre-existing
circulating antibodies is warranted.
39
THE IMPACT OF BLOOD PRESSURE CONTROL TOWARD
THE OUTCOME OF HEART TRANSPLANT RECIPIENTS
Moradzadeh N, Kawano M, Goldstein Z, Hamilton M, Kobashigawa J
Cedars-Sinai Heart Institute, Beverly Hills, CA.
Purpose of Study: Hypertension is a known risk factor for patients with
cardiovascular disease. It is not known to have the same impact in patients
post heart transplant. The immunosuppressive medications such as calcineurin inhibitors increase blood pressure (BP) and cause kidney injury, which
can further complicate the hypertensive state. BP measurements in the heart
transplant clinics may not be reliable as patients are told to hold their medication to obtain trough levels. Subsequently, this causes patients_ BP to rise
when they are seen in clinic. Therefore, we sought to determine the association between the number of BP medications and outcomes after heart
transplantation. We further assessed whether there was a protective effect
from any single BP medication as has been reported for ACE inhibitors in
heart transplant patients.
Methods Used: We evaluated 558 patients transplanted between 1994 and
2008. Patients were divided into groups based on the number of BP medications taken for Q 6 months within the first 2 years post heart transplant.
Control patients on 0 BP medications were selected for 2-year conditional
survival. Assessed 5-year subsequent outcomes included survival, freedom
from cardiac allograft vasculopathy (CAV, stenosis Q30%), and freedom from
non-fatal major adverse cardiac events (NF-MACE, MI, heart failure, PTCA,
pacemaker, stroke, new peripheral vascular disease). BP medications were
divided into several categories, including beta blockers, calcium channel
blockers, ACE Inhibitors, and angiotensin II receptor blockers.
Summary of Results: Patients with 3 BP medications exhibited a trend
toward lower survival compared to patients with 0 BP medications (70% vs.
82%, p=0.06) and 1 (70% vs. 82%, p=0.07) or 2 (70% vs. 85%, p=0.06) BP
medications. Subsequent freedom from CAV and NF-MACE were similar
between all groups. Over half of the patients maintained the antihypertensive
medication for the subsequent 5-year follow up. There was no difference in
outcome for the use of any particular class of BP medication in terms of long
term outcomes. Specifically, ACE inhibitors did not reduce angiographic
CAV as intimated in previous reports.
Conclusions: 3 or more BP medications are associated with poor outcome
post heart transplant. This suggests that more refractory hypertension may
relate to lower survival.
40
THE OUTCOME OF AFRICAN AMERICAN HEART
TRANSPLANT RECIPIENTS: ARE THEY TRULY AT RISK?
Geskin A, Kawano M, Goldstein Z, Hamilton M, Kobashigawa J CedarsSinai Heart Institute, Beverly Hills, CA.
Purpose of Study: African American heart transplant patients have been
reported to have increased risk of rejection due to different metabolism of
immunosuppressive medications. For example, differences in the activity of
the enzyme thiopurine methyltransferase which alters metabolism of such
drugs such as azathioprine, has been linked to ethnic disparity in clinical
outcome after heart transplantation. We reviewed our experience with African
American patients to confirm higher risk and difference in immunoresponsive
state in heart transplant patients on triple immunosuppressants.
Methods Used: We reviewed 1160 patients transplanted between 1994 and
2010, and divided them into racial groups (Caucasian, African American,
Latino, and Asian). The end points for this study included 10-year actuarial
survival, freedom from allograft vasculopathy (CAV, stenosis Q30%), freedom from non-fatal major adverse cardiac events (NF-MACE, MI, heart
failure, PTCA, pacemaker, stroke, new peripheral vascular disease), and first-
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
year freedom from any-treated rejection. Patients were treated on triple-drug
immunosuppression.
Summary of Results: The African American group had significantly
greater first-year freedom from cellular rejection compared to the Caucasian
group (89% vs. 94%, p=0.010). The African American group also demonstrated a trend towards lower first-year freedom from any-treated rejection in
comparison to the Caucasian group (85% vs. 91%, p=0.09). In addition, the
African American group demonstrated significantly lower 10-year freedom
from NF-MACE when compared to the Caucasian group (81% vs. 87%,
p=0.41) and the Asian group (81% vs. 92%, p=0.41). However, there was
no difference in 10-year survival and freedom from CAV.
Conclusions: African American heart transplant patients have a significantly higher first-year freedom from cellular rejection, a trend toward lower
first-year freedom from any-treated rejection, and significantly greater lower
10-year freedom from NF-MACE. Further study into metabolic differences
causing these findings is warranted.
41
THE LONG-TERM BENEFIT OF RENAL SPARING
PROTOCOLS DEMONSTRATE THE LACK OF NEED FOR
CALCINEURIN INHIBITORS
Dyo J, Kawano M, Goldstein Z, Hamilton M, Kobashigawa J Cedars-Sinai
Heart Institute, Beverly Hills, CA.
Purpose of Study: Calcineurin inhibitors (CNI) have been the mainstay of
immunosuppression after heart transplantation. However, these agents
have many adverse events which include kidney failure, hypertension,
hyperlipidemia, osteoporosis, and gout. There have been many attempts to
wean patients off CNI in patients who have developed ongoing renal failure.
There is concern that patients weaned off CNI may not have optimal long
term outcome. Therefore, we assessed our patients on a renal sparing protocol
(RSP) without CNI for long-term outcome compared to controls maintained
on CNI-based immunosuppression.
Methods Used: We reviewed 1266 patients transplanted between 1994 and
2010, and identified 40 patients on renal sparing protocol after heart transplantation. These patients were compared to a 2:1 contemporaneous control
group matched for age, gender, and time after transplantation (compared to
start of RSP). Subsequent five year outcomes including renal function,
actuarial survival, actuarial freedom from cardiac allograft vasculopathy
(CAV, stenosis Q30%), and actuarial freedom from non-fatal major adverse
cardiac events (NF-MACE, MI, heart failure, PTCA, pacemaker, stroke, new
peripheral vascular disease) were assessed. Patients placed on dialysis were
censored thereafter.
Summary of Results: Patients were found to start RSP at 5.5 T 3.1 years
post heart transplant, and had an average follow up time of 2.0 T 2.0 years.
Renal function improved significantly in those patients on the RSP compared
to those control not on RSP (percentage change in creatinine: j21% vs. 24%,
pG0.001). Subsequent five year actuarial survival, freedom from CAV, and
freedom from NF-MACE were found to be similar between the two
groups. 4/40 patients who started RSP eventually developed renal failure
requiring chronic renal dialysis at a mean time of 2.1 T 1.4 years post
RSP initiation.
Conclusions: A renal sparing protocol with withdrawal of calcineurin
inhibitors appears to be effective in preserving kidney function in the long
term without increase in mortality or morbidity (worsening renal functioning
necessitating kidney dialysis).
42
DOES GENDER MISMATCH INCREASE THE RISK OF
ANTIBODY-MEDIATED REJECTION?
Barry O, Kawano M, Goldstein Z, Hamilton M, Kobashigawa J CedarsSinai Heart Institute, Beverly Hills, CA.
Purpose of Study: The risk of antibody-mediated rejection (AMR) in heart
transplant patients is increased in multiparous females due to sensitization at
the birth of the baby. Sensitization can also occur due to blood transfusions,
ventricular assist device placement, and previous organ transplantation.
Recently, through several registries, it has been found that gender mismatch
(female donor-to-male recipient or male donor-to-female recipient) have
lower survival. The cause and mechanism of this finding has not been
elucidated. Since gender mismatch may enhance an immune response, we
choose to assess a risk for increased AMR.
99
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Methods Used: We assessed our gender matched and mismatched patients
transplanted between 1994 and 2010 to assess the incidence of 1st-year
AMR. These patients were divided into 4 groups based upon donor and
recipient gender (donor/recipient). AMR diagnosis was defined as pathologic, histologic and immunopathologic findings. We divided these patients
into asymptomatic AMR vs. treated AMR.
Summary of Results: We found that the incidence of 1st-year AMR was
significantly increased in the M/F group versus M/M (11% vs. 5%, p=0.005),
and F/F (11% vs. 6%, p=0.046). The M/F group was found to have significantly greater asymptomatic AMR compared to the M/M group (6% vs. 2%,
p=0.016) but not to the F/F group. The F/M group was found to have similar
incidences of 1st-year treated AMR and asymptomatic AMR when compared
to the gender matched groups. In the M/F and F/F groups, there was no
difference in the percentage of multiparous females.
Conclusions: Heart transplant patients with M/F gender mismatch on tripledrug immunosuppression have an increased incidence of 1st-year antibodymediated rejection compared to gender match patients. Although multiparous
female recipients are known to have more AMR, the F/F gender match group
had similar incidence of AMR to those of the M/M gender match group.
Future efforts to understand and attenuate the underlying mechanisms are
essential.
43
THE DEVELOPMENT OF LATE ONSET AMR: DOES
IT EXIST?
Goldstein Z, Kawano M, Hamilton M, Kobashigawa J Cedars-Sinai Heart
Institute, Beverly Hills, CA.
Purpose of Study: Antibody-mediated rejection (AMR) is usually seen
early after heart transplantation and may be associated with graft dysfunction
and circulating antibodies. Interestingly, as the risk of cellular rejection
gradually decreases the incidence of AMR appears to be rising. Late onset
AMR is also becoming more prevalent. We choose to assess our patients
with late onset AMR (greater than one year) and characterize their clinical
presentation.
Methods Used: We reviewed 1290 patients transplanted between 1994 and
2010 for the development of AMR and divided them into groups based upon
diagnoses of less than one year and greater than one year following heart
transplantation. The definition of AMR was based on pathology of heart
biopsy including characteristic histology and immunopathology findings.
These patients were then assessed for the development of restrictive physiology, hemodynamic compromise, and subsequent 5-year outcomes including actuarial survival and the development of transplant coronary artery
disease (stenosis Q30%).
Summary of Results: There were 112 patients in the early AMR group
versus 37 patients in the late AMR group. There was a greater trend for the
development of restrictive physiology (10.7% vs. 2.5%, p=0.10) and hemodynamic compromise (17.1 % vs. 9.4%, p= 0.19) for the late AMR group.
Similarly, those patients exhibiting late AMR also demonstrated a greater
trend towards developing subsequent 5-year transplant coronary artery disease (43.2% vs. 28.3%, p=0.06). Treatment of these AMR episodes included
IV Solumedrol, Rituximab, Plasmapheresis, Bortezomib, ATG, or IVIG, with
no significant difference in subsequent 5-year actuarial survival between
the two groups (65.2% vs. 54.1%, p=0.285).
Conclusions: Late antibody-mediated rejection demonstrates a consistent
trend towards more severe presentation with hemodynamic compromise,
increased risk for developing CAV, and increased risk in the development of
restrictive cardiac physiology. Continued surveillance for the detection and
means for prevention of AMR in heart transplant patients late after transplant
should be pursued.
44
30 DAY MORTALITY AND ACUTE READMISSION
FOLLOWING PERCUTANEOUS CORONARY
INTERVENTION IN A COMMUNITY SETTING
Blakley B2, Daratha K1,2, Salvatierra G1 1Washington State University,
Spokane, WA and 2University of Washington, Seattle, WA.
Purpose of Study: To establish 30 day mortality and acute readmission
rates following percutaneous coronary intervention (PCI). Previous studies
examining 30 day mortality and acute readmission following PCI have been
limited to Medicare populations and to university-based medical centers.
100
&
Volume 59, Number 1, January 2011
Our study was designed to include all payers in a community multi-hospital
setting.
Methods Used: Analyses included mortality and all inpatient readmissions
including: cardiac, non-cardiac, and all cause, following PCI. There were
9,650 cases (68.8% male) with an average age of 65.5 at their index procedure. The study included cases from patients first treated with PCI
at Providence Sacred Heart Medical Center (PSHMC) between 3/1/2001 and
3/31/2006, followed through 12/31/2009. The study population in this
analysis was extracted from the Spokane Community Clinical Data Repository (SCCDR), currently populated from three sources: Apollo (identifying
revascularization information), Meditech (identifies readmission information), and the Social Security Administration Death Index (identifies death
dates). Only primary ICD-9 diagnosis codes indicating cardiac or non-cardiac
readmission were used to determine readmission causes.
Summary of Results: The overall 30 day mortality rate in this study was
1.5% and overall 30 day readmission rate was 11.1%. Patients Q65 showed a
mortality rate of 2.2%, compared to patients G65 with a mortality rate of
0.7% (pG.001). Males showed a mortality rate of 1.2%, females had a mortality rate of 2.2% (pG.001). Following PCI, males Q65 had significantly
lower mortality rates compared to females Q65, 1.7 vs. 3.1% (p=.002), while
men and women e65 did not have significantly different mortality rates 0.8%
(p=.388). The comparison of male to female all cause readmission was 9.7 vs.
13.3% (p=.001) and cardiac readmissions were 8.2 vs. 10.8% (p=.006)
respectively for the G65 age groups.
Conclusions: This study reports 30 day mortality and readmission rates at
1.5% and 11.1% respectively. Differences between this data and previous
studies are attributed to a younger population, and from including a community population which previous studies, based exclusively in universitybased medical centers, were unable to attain.
45
IMPACT OF METABOLIC SYNDROME ON
CARDIOVASCULAR AND MENTAL DISEASES
IN A VA POPULATION
Singh M2,1, Gu W1,2, Mallios R1, Mcfarland S1, Fong J1, Huang J1,2
1
VA Central California Healthcare System, FRESNO, CA and 2UCSF,
Fresno, Fresno, CA.
Purpose of Study: Metabolic syndrome (METS) is a cluster of conditions
that confer greater cardiovascular (CV) risk and may be associated with
certain mental disorders. A quarter of US population is currently affected by
METS and the prevalence increases with age. We sought to examine the
prevalence of METS and to determine the impact of METS on CV and
psychiatric diseases in a VA population.
Methods Used: Demographics and clinical data on diagnosis, labs and
medication profiles were collected from 17,466 patients in this crosssectional study. METS was defined by modified NCEP criteria. T -test and
Chi square test were used for demographic and risk comparison between
those with and without METS. Chi square test and ANOVA were used for risk
comparison among groups with increasing number of METS components.
Logistic regression model was used to determine the adjusted odds ratios.
Summary of Results: Mean age was 67 years with 96 % males. Prevalence
of METS was 58%. Although numbers of patients who ever smoked tobacco
between the two groups were similar (P90.05), there were significantly less
current smokers in METS+ group (PG0.001). Among patients with METS,
there were significantly higher mean FS and increased prevalence of PCI, MI,
CABG, PVD, ED CKD, depression, PTSD (PG0.001), CVA/TIA, and anxiety
(PG0.002). TC and LDL were lower while non-HDL cholesterol was higher in
METS+ group, consistent with more atherogenic lipid profiles. There was
significantly proportional increase in a number of CV and mental diseases
with increasing number of METS components. After adjusting for age, sex,
BMI, diagnosis of hypertension and diabetes, and statin use by logistic
regression model, METS remained significantly associated with MI, CABG,
PVD, CKD, anxiety and PTSD.
Conclusions: There are more than double cases of METS in this study
compared to US general population. METS is significantly associated with
certain CV and mental diseases and the degree of this association becomes
stronger with increasing number of individual METS components. More
aggressive intervention targeting multiple CV risk factors is particularly
important for this high-risk patient population. Our results also suggest the
need to reduce concomitant CV risk in the long-term management of
depression, anxiety, and PTSD.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
46
INCIDENCE, ETIOLOGY, AND SURVIVAL TRENDS FROM
SUDDEN CARDIAC ARREST IN CHILDREN AND
YOUNG ADULTS AGE 0-35 IN KING COUNTY,
WASHINGTON: A 30-YEAR REVIEW
Meyer L1, Drezner J1, Fahrenbruch C2, Stubbs B2, Maeda C1, Harmon K1,
Eisenberg M1,2 1University of Washington, Seattle, WA and 2King County
Emergency Medical Services, Seattle, WA.
Purpose of Study: Sudden cardiac arrest is a leading cause of death in
children and young adults, although little is known regarding the epidemiology of such events. This study determined the incidence, etiology, and
outcomes of out-of-hospital cardiac arrest (OHCA) in individuals age 0Y35.
Methods Used: This was a retrospective cohort study of OHCA in children
and young adults from January 1980 through December 2009 in King
County, Washington. Cases of OHCA were identified from the King County
Public Health Division of Emergency Medical Services (EMS) Cardiac
Arrest Database, an ongoing registry of all cases of OHCA involving an EMS
response. Incidence rates were calculated using population census data, and
the etiology of cardiac arrest determined by review of autopsy reports, death
certificates, hospital and other available records.
Summary of Results: A total of 361 cases (26 cases age 0Y2, 30 cases age
3Y13, 60 cases age 14Y24, and 245 cases age 25Y35) of OHCA occurred,
with an overall incidence of 2.28 per 100,000 person-years (2.1 in age 0Y2,
0.61 in age 3Y13, 1.44 in age 14Y24, and 4.40 in age 25Y35). The most
common etiologies of OHCA were congenital abnormalities (84.0%) in
age 0Y2, hypertrophic cardiomyopathy (17.9%) in age 3Y13, presumed
primary arrhythmia (23.5%) in age 14Y24, and coronary artery disease
(42.9%) in age 25Y35. The overall survival rate was 26.9% (3.8% in
age 0Y2, 40.0% in age 3Y13, 36.7% in age 14Y24, and 27.8% in age 25Y35).
Survival increased significantly throughout the study time period from
13.0% in 1980Y1989 to 40.2% in 2000Y2009 (pG0.001).
Conclusions: The incidence of OHCA in children and young adults is
substantially higher than previously reported. This study details the various
cardiac disorders leading to OHCA in the young, allowing an increased focus
on age related risk factors and development of effective screening models
for prevention. This study demonstrates an important increase in survival,
providing compelling support of contemporary resuscitation protocols for
OHCA in the young.
47
EFFECT OF RADIOFREQUENCY ABLATION OF
VENTRICULAR TACHYCARDIA ON EJECTION FRACTION
IN PATIENTS WITH CARDIOMYOPATHY
Finch W, Vaseghi M, Shivkumar K David Geffen School of Medicine at
UCLA, Los Angeles, CA.
Purpose of Study: To determine the effect of radiofrequency ablation
(RFA) for ventricular tachycardia (VT) on ejection fraction (EF) and the
correlation of the change in EF with the number of radiofrequency lesions
applied.
Methods Used: We performed a retrospective chart review to create a
database of patients who underwent radiofrequency ablation for VT at UCLA
since 2002. A total of 125 patients received 153 procedures. The average age
of patients in the database was 57 T 16. Of the 125 patients, 84 were male, and
41 were female. The majority of patients (92) had cardiomyopathy; 43 had
non-ischemic cardiomyopathy, and 49 had ischemic cardiomyopathy. Parameters included in the database were the pre-procedure and post-procedure
EF and the number of RFA lesions applied during the procedure, as well as
other procedural characteristics and outcome data. The Wilcoxon signed-rank
test was performed to compare the pre-ablation and post-ablation EF, as the
data did not follow a normal distribution. The correlation coefficient between
the number of RFA lesions and the change in EF ($EF) was calculated.
Summary of Results: The median of both pre-ablation and post-ablation
EF = 27.5 (p = 0.15). The correlation coefficient between the number of RFA
lesions and $EF was -0.08.
Conclusions: Although it has historically been thought that because
radiofrequency creates burns in myocardium, ejection fraction would worsen
after VT ablation, the present study determines that this is not the case. There
was no significant change in EF following ablation. Additionally, there was
no correlation between the number of radiofrequency lesions and $EF. These
results confirm findings from previous studies that VT ablation is not detri-
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
mental to cardiac function, and that applying more lesions does not result
in worsened EF.
48
CHANGES IN SURVIVAL FROM PULSELESS ELECTRICAL
ACTIVITY OR ASYSTOLE CARDIAC ARREST IN KING
COUNTY, WA BEFORE AND AFTER IMPLEMENTATION
OF THE 2005 AMERICAN HEART ASSOCIATION
RESUSCITATION GUIDELINES
Redshaw JD1, Kudenchuk PJ1,2, Rea TD1,2, Fahrenbruch C2, Stubbs B2,
Eisenberg M1,2, Phelps R1,2 1University of Washington, Seattle, WA and
2
Seattle and King County Public Health, Seattle, WA.
Purpose of Study: Increased survival from ventricular fibrillation (VF) out
of hospital cardiac arrest (OOHCA) has been reported in several communities
following implementation of the 2005 American Heart Association (AHA)
resuscitation guidelines. No study to date, however, has focused specifically on the impact of the 2005 AHA guidelines on OOHCA in cases with
pulseless electrical activity (PEA) or asystole as the initial rhythm. We
evaluated whether the re-emphasis on high quality, minimally interrupted
CPR as outlined in the 2005 guidelines increased survival in patients in
PEA or asystole arrest.
Methods Used: The investigation was a retrospective cohort study of all
EMS treated persons with non-traumatic OOHCA and a non-shockable initial
rhythm between 1/1/2000 and 3/31/2010 in King County, WA. A 5 year
historical control was compared to a 5 year intervention group after the
protocol change. The primary outcome was survival to hospital discharge.
Secondary outcomes included return of spontaneous circulation, discharge
location, and CPC score. Contingency tables and multivariable logistic
regression models were used to assess these outcomes before and after the
protocol change.
Summary of Results: 1780 patients were included in the control group and
2173 in the intervention group. Patient demographics, first response time,
proportion of witnessed arrests, and provision of ALS care were similar
between the groups. Unadjusted OOHCA survival rates were 4.4% in the
control period and 6.1% in the intervention period (odds ratio [OR] 1.39;
95% confidence interval [CI] 1.05 to 1.86). In the subgroup of witnessed
arrests, survival was 6.4% in the control and 10.4% in the intervention period
(OR 1.71; 95% CI 1.17 to 2.50). Multivariate regression analysis adjusting
for significant predictors of survival was associated with 1.72 greater odds
of survival from witnessed OOHCA in the intervention period (95% CI 1.15
to 2.58).
Conclusions: In King County, WA, OOHCA survival rates improved in
patients initially found in a non-shockable rhythm after implementation of a
minimally interrupted CPR protocol.
Community Health
Concurrent Session
8:30 AM
Friday, January 28, 2011
49
PROMOTING THE INTEREST OF UNDER-REPRESENTED
YOUTH IN HEALTH CARE THROUGH ONE-ON-ONE
MENTORSHIP
Arellano D, Ebneshahrashoob M, Saldana L, Fogel A, Afghani B University
of California, Irvine, Irvine, CA.
Purpose of Study: Under-representation of minority groups in the health
care field continues to be a major problem. Lack of mentors and role models
have attributed to under-representation of minority groups in health care
fields. The objective of this study was to evaluate the effect of a one-on-one
mentorship program in promoting the interest of underserved high school
students in healthcare fields.
Methods Used: At the Center for Future Health Professionals at UC Irvine,
ten undergraduate or postgraduate mentors were matched with 10 minority
high school students based on their interests. During the one year period, the
students worked on a project that represented a health care challenge in the
student_s local community. The high school students (mentees) chose and
researched the topic under the supervision of their mentor. The projects were
presented at the end of the year in a gathering.
101
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Summary of Results: Health care projects chosen by the mentees included
teenage pregnancy, alcoholism, obesity, heart disease, schitzophrenia and
smoking. The mentees evaluated the program as very effective in building a
bond with their mentor, becoming aware of community problems as well
as increasing their knowledge and interests about different health-related
careers. The mentors liked the sense of giving back to the community and
accessibility to the directors of the program. Both the mentors and the
mentees said that the flexibility in scheduling and pairing according to their
interest was very helpful. The mentees stated that they wished they had
more time with the mentors.
Conclusions: Our one-on-one mentorship program which matched the
mentors and mentees based on their interest and empowered the mentees to
lead a project was very effective and raised both the mentee and mentors_
interest in healthcare fields and community service. Because of the interest of
our school partners, students and mentors, our program is expanding rapidly.
50
IDENTIFYING FACTORS IN UNDER-REPRESENTATION
OF MINORITY YOUTH IN HEALTHCARE CAREERS
Afghani B, Saldana L, Arellano D, Fogel A, Ebneshahrashoob M University
of California, Irvine, CA.
Purpose of Study: The under-representation of minority population in
different healthcare fields has been identified as one of the major reasons
responsible for healthcare disparity in the United States. The objective of this
study was to: a) evaluate some of the barriers among high school students in
choosing healthcare careers and b) to assess the effectiveness of our workshops in introducing different healthcare professions to high school students.
Methods Used: At the Center for Future Health Professionals, UC Irvine,
we conducted workshops regarding different healthcare fields in several high
schools with significant drop-out rates. We distributed a questionnaire to
evaluate the barriers in choosing healthcare fields as a career.
Summary of Results: One hundred and ninety students were surveyed.
The response rate was 100%. Of 190 students 85% were Hispanic. The most
common barriers identified in choosing a healthcare field included lack of a
role model, lack of financial support and the long road to finish. Majority of
students were not aware of careers in allied health care fields, such as child
life, social work, respiratory therapy, case management and healthcare billing.
The students evaluated our workshop as very effective in introducing them to
different healthcare professions and more than 90% said that they were more
interested in a healthcare field as a result of our workshops.
Conclusions: Among several barriers, lack of knowledge about different
healthcare careers may contribute to the under-representation of minority
groups in healthcare fields. Programs that address the barriers responsible for
under-representation of minority in healthcare are needed.
51
ADDRESSING CHILDHOOD OBESITY THROUGH THE
DEVELOPMENT OF A FARMERS_ MARKET IN
GILLETTE, WYOMING
Gorden N University of Washington, Seattle, WA.
Purpose of Study: Campbell County in northeastern Wyoming is experiencing the state_s largest population growth rate, particularly among persons
under age 18. Despite increased spending on recreation programs and high
participation in school athletics, the school district shows that 22% of boys
(N=886) and 22.6% of girls (N=823) in grades 7Y9 are in the 95th percentile
for body mass index and are obese. Access to healthy food, one factor
associated with childhood obesity, is poor. The goal was to establish a
farmers_ market (FM) to provide a source of fresh fruit and vegetables.
Methods Used: Discussions with the school health director, public health
officer, child development coordinator, and physicians revealed childhood
obesity as a health concern. Meetings with local food producers, city officials,
and volunteers were aimed at increasing FM attendance and attracting diverse
food vendors. Research of peer-reviewed articles on obesity and nutrition
identified barriers to healthy foods and effective methods of teaching nutrition. At the FM, media was distributed with the USDA diet guidelines and
listing the benefits of fruit and vegetable consumption. School-aged children
were allowed to try new fruit and vegetables. FM visitors and state representatives received WIC and Senior Farmers_ Market Nutrition Program
resources to bring awareness to federal food subsidy programs.
102
&
Volume 59, Number 1, January 2011
Summary of Results: The FM was moved to a new location to accommodate growth. Ten vendors participated in the first market of the 2010
season: 4 local farmers, 2 confectioners, 1 jeweler, and 3 educators. In four
hours, over 150 visitors attended. Sixteen school-aged children sampled fruit
and vegetables and 20 visitors accepted brochures on the benefits of consuming local fruit and vegetables. Seven visitors inquired about federal food
subsidies. By the seventh week of the FM, 19 vendors and 579 visitors
attended.
Conclusions: Research suggests that individual-centered interventions are
insufficient to address childhood obesity and related disease. Corrective
action must focus on the home, community, and national environment. The
atmosphere of FMs can reach this broad scale. Further FM growth will have
an additive effect on the overall health of communities like Gillette, WY
which already boast high levels of physical activity and a steady economy.
52
PREVENTION OF POSTPARTUM SMOKING RELAPSE IN
MOTHERS TO PREVENT INFANT EXPOSURE TO
SECOND-HAND SMOKE
Adams KK, Merritt TA Loma Linda University, Loma Linda, CA.
Purpose of Study: A significant number of women quit smoking during
pregnancy; however, a majority of them resume smoking following delivery.
Our study aims to prevent postpartum smoking relapse during the first
8 weeks postpartum through a weekly educational messaging program and
thus reduce infants’ exposure to second-hand smoke.
Methods Used: This project was a prospective randomized trial. Mothers of
infants delivered at Loma Linda University Medical Center were interviewed
during their postpartum stay regarding smoking history. Mothers who
smoked during the 12 months prior to conception or during pregnancy and
who were not currently smoking were randomized to the trial. Mothers were
randomized to a Standard of Care (SOC) or Smoking Relapse Prevention
(SRP) group. Mothers in both groups watched an educational video while in
the hospital regarding the importance of smoking cessation. Mothers in the
SRP group received additional information after discharge, including
biweekly robotic phone calls, weekly mailings, and emails which provided
education on smoking relapse prevention and normal newborn care. All
mothers were contacted at 2 months postpartum to assess smoking status.
Summary of Results: From March 2010 to September 2010, 328 mothers
were interviewed. Twenty-nine mothers were identified as smokers, of which,
5 mothers were excluded or declined to participate. To date, 12 mothers have
been contacted at 2 months postpartum, and follow-up information was
available for 8. All mothers identified the video as effective. No mother
identified calling as a useful means of follow-up communication. Two
mothers (25%) reported smoking relapse, 1 in the SOC group and 1 in the
SRP group. There are 12 additional mothers who will contacted by the end
of October 2010.
Conclusions: The postpartum stay provides an opportune time to teach
mothers regarding the importance of smoking cessation and to maintain a
smoke free environment for their infants. Extended contacts through educational messaging may not be enough to prevent smoking relapse.
53
DEPRESSION IN YOUTH: THE ROLE OF PASTORAL CARE
IN BUTTE, MT
Kwan A University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Butte is a low-income town of approximately 34,000
people located in southwest Montana with an extremely high incidence of
depression and suicide. The purpose of this project is threefold: To raise
awareness of the significant problem that depression represents in the community, to decrease stigma associated with mental health issues, and to
provide important community figures with the ability to recognize depression
in children and adolescents, and referral resources for local mental health
providers.
Methods Used: Depression was identified as a major issue through input
from local healthcare providers. Research and literature review were performed to specify the target population and intervention method. Through
community assistance by the chaplain at the local hospital, individual
meetings were arranged with leaders representing Catholic, Lutheran,
Serbian Orthodox, and Episcopal faiths. The religious leaders were engaged in conversations regarding the prevalence of mental health issues in
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
their community, and their experiences with and their attributions for
depression. They received a brief education on identifying depression in
youth, were provided with materials covering ways pastoral care can help,
and were given a list of community mental health resources for referral,
including a personal line to a case manager with experience in dealing with
youth mental health issues.
Summary of Results: The educational material was generally very well
received, with the acknowledgement that both the identification methods and
the referral information were useful. Most of the religious leaders had
experience with depression and suicide in the community, whether through
pastoral care, funeral services, or personal experience. Attributions for the
community prevalence of depression included: alcoholism, secularism,
general depressed state of the town, lack of community and social support
structures, and prevalence of abuse.
Conclusions: Engagement of community leaders regarding mental health
issues and resources allowed for the opportunity to increase awareness of the
issues facing the community, the signs and symptoms of child and adolescent
depression, and the available community mental health resources. Face-toface meetings allowed for a personal and open connection to occur, which
may have increased benefit for reception of the material.
54
ENGAGING CHILDREN IN PHYSICAL ACTIVITY AND
HEALTH AWARENESS, SEATTLE WA
Le TH University of Washington School of Medicine, Seattle, WA.
Purpose of Study: The Rainier Valley has the greatest cultural and
socioeconomic diversity in Washington State, but nearly twice the poverty
rate of the greater Seattle area. This inequality has a profound effect on
childhood development: Children from low-income families are more likely
to become obese, which leads to later chronic health problems. These children are also 80% less likely to receive routine health and vision care. The
purpose of this project was to promote exercise and connect families to
resources for free vision care in the community.
Methods Used: Sports-based youth development programs are strongly
supported in the literature for positive influence on behavior. This project
utilized a tennis camp to advocate both physical activity and to distribute
information regarding health and vision care. With support from the community, donations and volunteers were gathered to organize a non-profit
tennis camp for children. Participants received individual tennis instruction
from volunteer coaches and were encouraged to continue exercising. At the
end of tennis camp, each participant also received: 1) A new junior tennis
racquet and balls. 2) Information about local tennis facilities. 3) NeighborCare Health\ medical clinic brochures. 4) A letter addressed to parents/
guardians about the importance of routine health and vision care. 5) Information about how to access Sight for Students, a national charity program for
free eye exams and glasses for students under 18.
Summary of Results: Eight children, ages 8Y12, attended both days of
tennis camp and received new, donated equipment. Participants and their
families expressed appreciation for the event and interest to continue playing
tennis. A community outreach letter, including a consolidated information
sheet about Sight for Students was created and distributed to twelve families
during the event. This information was reproduced and made available at the
Rainier Park Medical Clinic.
Conclusions: Sports-based youth development programs involve extensive
coordination and organization, and are limited by the number of participants.
Although it may not be the most efficient intervention, the experience
for each participant is powerful and lasting. To maximize community outreach, health information that is well-organized and easy for patients to
understand should also be distributed in these programs and reproduced for
future use.
55
REDUCING FRUSTRATION TO INFANT CRYING: A LAB
BASED TEST OF THE PERIOD OF PURPLE CRYING
MATERIALS
Lou C1, D’Souza N2, Chen M2, Barr R2 1University of British Columbia,
Coquitlam, BC, Canada and 2Child and Family Research Institute,
University of British Columbia, Vancouver, BC, Canada.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Purpose of Study: Infant crying is the most common stimulus for Shaken
Baby Syndrome (SBS) that results in traumatic brain injury in infants. The
Period of PURPLE Crying is a SBS prevention program that is implemented
jurisdiction-wide in British Columbia, Canada and many states in the USA. It
aims to reduce SBS incidence by informing new parents about the properties
of crying in normal infants, the dangers of shaking, and coping strategies.
The object of this study is to determine whether the Period of PURPLE
Crying DVD reduces maternal frustration and affects choice of coping
strategies in response to prolonged crying.
Methods Used: Participants in this study are first time pregnant women
fluent in English with no previous exposure to PURPLE. In a randomized
controlled trial, participants watched either the PURPLE DVD or a control
DVD on Infant Safety. Following the DVD, participants listened to a
10-minute audio-recording of an infant crying (duration unknown to participants) while continuously rating frustration levels on a visual analog
scale (CVAS). Scores ranged from 0 to 100 (Fno frustration_ to Fextreme
frustration_). Participants completed a post test questionnaire asking how
likely (from 0=’not at all likely’ to 4= ’extremely likely’) they were to use
each of ten strategies to cope with crying.
Summary of Results: To date, 33 participants (19 PURPLE & 14 control)
have participated. Mean frustration level during the audiorecording was lower
in the PURPLE than in the control group (19.3T15.5 vs. 28.5 T 23.4, t=j1.3;
p= 0.10). PURPLE participants reported greater likelihood to Bput baby
somewhere safe and walk away[ as a coping strategy (t=2.1, p=0.04). There
were no significant differences in employing the other nine strategies.
Conclusions: PURPLE participants responded to infant crying with lower
frustration than controls. They were also more likely to choose a Bwalk away[
strategy than controls. These preliminary results suggest that PURPLE
education materials may have an impact on maternal frustration and coping
behaviors considered relevant to preventing shaking. Data collection will
continue until the predetermined sample size (n=45/group) is reached.
56
ORAL HYGIENE EDUCATION FOR OLDER CHILDREN IN
CENTRALIA, WASHINGTON
Srivastava K University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Tooth decay and infection are common in children at the
Pope_s Kids Place Medical Clinic in Centralia, WA. The majority of patients
are of low-income and have DSHS (Medicaid) insurance; as a result, they are
at a higher risk of developing tooth decay. The Pope_s Kids Place Dental
Clinic is part of Washington_s Access to Baby and Child Dentistry Program
through DSHS. The ABCD program provides dental education and prevention for children from birth to six years of age. However, there is no outreach
program for children over six years of age at the dental clinic. The purpose of
this project was to develop a presentation about preventing tooth decay
through healthy oral hygiene practices, with the target population of eight to
ten year olds.
Methods Used: Discussions with the dental clinic education/outreach
coordinator led to identifying the target population and development of
materials for the presentation. Four main points were identified for the presentation based on the observations of the coordinator educating children and
parents on oral hygiene. A literature review was conducted to validate and
support the main ways to reduce the risk of developing dental caries in
children - low sugar diet, tooth cleansing, fluoridation, and regular dental
visits.
Summary of Results: The presentation was given to 35 elementary summer school students. First, children brushed off grape juice from hard-boiled
eggs to simulate brushing of the teeth to remove sugars. Proper brushing and
flossing techniques were demonstrated on a large mouth model. Finally, a
laminated flyer for parents was distributed containing tips for maintaining
healthy teeth and dental clinic referrals on one side and a brushing/flossing
daily tracking chart on the other side. The children and their teachers were
very receptive to the presentation. The dental clinic can use this presentation
as part of their outreach program in the future.
Conclusions: The goal of the project was to educate older children on
prevention of dental disease and instill good oral health practices, and in turn,
the children can inform parents and younger siblings of their education.
Pediatricians are the first healthcare providers that children see and therefore,
pediatricians can be actively involved in identifying tooth decay and educating families in preventing infection in the future.
103
Western Regional Meeting Abstracts
Journal of Investigative Medicine
57
ADOLESCENT SUICIDE PREVENTION IN DAYTON,
WASHINGTON
Wurz KA University of Washington, Seattle, WA.
Purpose of Study: Dayton is a small town, population 2,735, located within
Columbia County in the southeastern corner of Washington. Adolescents
make up roughly 10 percent of Dayton_s population, for whom suicide is a
serious public health concern. Suicide is the third leading cause of death for
adolescence in the United States and in 2008 16% of Columbia County youth
reported having considered suicide. Studies have shown that school-based
suicide prevention programs are an effective method for reducing adolescent
suicide rates. With the loss of several Dayton adolescents to suicide, the most
recent in June 2010, this serious public health issue must be addressed by the
community. The purpose of this project was to catalyze and communicate
with key youth community resource organizations to encourage appropriate
adolescent suicide prevention.
Methods Used: The need for a suicide education program in Dayton was
revealed by meeting with: adolescents, public health nurses, a mental health
preventionist, and the Columbia County Coalition for Youth and Families.
These groups collectively agreed that use of both brochures and an in-class
educational program would be most effective. A literature review was preformed to identify effective school-based suicide education programs and to
provide guidance in the creation of an educational flyer. A written report on
adolescent suicide and effective school-based prevention programs was
presented to several youth community resource organizations. The key
suggestion was purchase and implementation of the Signs of Suicide Prevention Program in Dayton schools.
Summary of Results: The mental health preventionist and Dayton School
District superintendent plan to meet and discuss suicide prevention before the
start of school in fall. An educational flyer was created and placed in Dayton
High School restrooms.
Conclusions: The community of Dayton now recognizes the need for an
adolescent suicide prevention program. Various community groups within
Dayton are now taking steps toward implementing a school-based prevention
program, but the cost will be a challenge. The use of flyers to educate Dayton
adolescents about the warning signs and risk factors of suicide is more
cost effective; unfortunately the impact of the flyers produced is not statistically known.
58
HEALTHY LIFESTYLE INTERVENTION: EDUCATING
FIFTH AND SIXTH GRADE STUDENTS WITH THE
HEALTH4LIFE PROGRAM
Young K1, Serrano V1, Castillo F2, Baum M 3 1Loma Linda University,
Loma Linda, CA; 2Loma Linda University, Loma Linda, CA and 3Loma
Linda University Medical Center, Loma Linda, CA.
Purpose of Study: Childhood obesity is listed in California as a significant
health issue. The Health4Life program was an eight week school-based
healthy living educational intervention conducted in fifth and sixth grade classrooms through May, 2010. The questions examined for this analysis were:
1. Did students_ knowledge on healthy living topics increase?
2. Did students increase in healthy behaviors over the program course?
Methods Used: The Health4Life program was conducted in 27 classrooms with a total of 1262 students enrolled. Weekly intervention consisted of a one-hour lecture on 1) Sleep, 2) Breakfast, 3) Soft Drinks, 4)
Fruits and Vegetables, 5) Feelings and Depression, 6) Fats/Fast Food 7) TV
Time/Exercise and 8) Safety. Pretests and post-tests were analyzed for
health knowledge. Individual weekly surveys were analyzed at week one
for baseline, week five and week eight for behavior change.
Summary of Results: Health knowledge improved as students showed a
9.9% increase (p=0.000) in comprehensive knowledge, and specifically in
the topics of Breakfast (11.8%), Soft Drinks (20%), Fats/Fast Food (12.5%),
TV Time/Exercise (13.1%) and Fruits and Vegetables (4.5%). Behavioral
change showed a significant decrease in Fats/Fast Food consumption from
baseline at week five (8.2%, p=0.006) and week eight (12.5%, p=0.011).
There was a decreasing trend in soda consumption at week five (3.8%) and
week 8 (4.3%). Of interest, 20% of students consistently reported weekly
episodes of sadness.
Conclusions: The Health4Life classroom based intervention demonstrated
a health benefit in knowledge acquisition and a change in behavior choices
104
&
Volume 59, Number 1, January 2011
with a decrease of fast food consumption over eight weeks. Additional
information could be gained from longitudinal follow up. Lifestyle changes
are difficult to sustain without additional parental involvement. A benefit to
the program could be Brefresher lectures,[ an incentive system for healthy
choices, and continued school participation in the areas of nutrition and
exercise.
59
STUDENT PERSPECTIVES ON THE INSTITUTIONAL
DIVERSITY CLIMATE AT A U.S. MEDICAL SCHOOL: THE
NEED FOR A BROADER DEFINITION OF DIVERSITY
Dhaliwal JS1, Crane LA2, Kilkenny R3, Valley MA1, Kaul P1, Nyquist A1,
Lowenstein SR1,2 1University of Colorado School of Medicine, Aurora, CO;
2
Colorado School of Public Health, Aurora, CO and 3Colorado Health
Foundation, Denver, CO.
Purpose of Study: Medical schools frequently experience problems related
to diversity and inclusiveness. The authors conducted this study to assess,
from the student body_s perspective, the climate at one medical school with
respect to diversity, inclusiveness and cross-cultural understanding.
Methods Used: In 2008, students in the M.D., P.T., and P.A. programs at a
medical school were asked to complete a survey consisting of 24 Likert-scale,
short-answer and open-ended questions. Questions were designed to measure
student experiences and attitudes in six domains: general environment and
culture; witnessed negative speech or behaviors; barriers to incident reporting; isolation of minorities; diversity and the learning environment; and
campus leadership.
Summary of Results: Of 883 eligible students, 261 (29.56%) participated.
Most respondents agreed that the SOM campus is friendly (90%) and welcoming to minority groups (82%). Although 90% found educational value in
a diverse faculty and student body, only 37% believed the campus is diverse.
Likewise, many respondents have witnessed students or residents make
disparaging remarks or exhibit hostile behaviors toward persons with strong
religious beliefs (43%), low socioeconomic status (35%), non-English
speakers (34%), women (30%), racial/ethnic minorities (28%), and GLBT
individuals (25%). Respondents witnessed similar behaviors from faculty
members toward persons with strong religious beliefs (18%), women (18%),
and persons of low socioeconomic status (12%). Open-ended comments
highlighted strong religious beliefs and conservative values as common targets. Students were unlikely to report these behaviors because of grading
concerns (95%) and the absence of a confidential reporting system (28%).
Conclusions: Diversity and a culture of inclusiveness are important medical
school assets. Schools must utilize broad definitions of diversity such that all
minority groups are valued, including those with conservative viewpoints
and strong religious beliefs.
60
AUDIENCE RESPONSE SYSTEM USE IN UNIVERSITY OF
BRITISH COLUMBIA MEDICAL SCHOOL
UNDERGRADUATE EDUCATION
Wilson I1, Afshar K1, Peterson L2, Masterson J1 1University of British
Columbia, Vancouver, BC, Canada and 2University of British Columbia,
Vancouver, BC, Canada.
Purpose of Study: To further explore the feasibility and benefits of the use
of an audience response system (ARS) in the UBC distributed MD undergraduate program, and to determine whether students favour an inquiry based
learning format incorporating ARS in the educational sessions following the
weekly problem based learning (PBL) case. Finally, to gather feedback from
instructors and students as to how to improve the use of ARS.
Methods Used: Students used an ARS device simultaneously at the
3 geographically separated medical program sites to answer 6 to 7 multiple
choice questions posed by instructors. Questions were related to a recently
completed PBL case and the week_s lecture content. Immediate feedback was
provided as a histogram representing all students_ answer choices and answer
options were discussed in detail. Students were surveyed twice using the
ARS. An e-mail survey of instructors was conducted. Also, a focus group
composed of medical students took place to gather opinions on the ARS.
A 5 point Likert scale was used.
Summary of Results: Students were surveyed in weeks 1 and 4 out of a
5 week period of study. The median answer was Bagree[ to all 5 survey
statements, demonstrating students (average n=180) favoured the educational
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
sessions using an ARS. After 4 weeks of use there was no change in the
response to questions 1 and 2 of the survey (Median=4 both week 1 and 4
Mann Whitney U test p=0.23). This was a uniform finding across the 3 sites.
All instructors (n=6) surveyed felt that use of the ARS should be expanded
within the undergrad curriculum.
Conclusions: The majority of students agreed that the introduction of an
ARS to post-PBL sessions aided them in identifying learning gaps, enhancing understanding of key concepts, and provided for an enjoyable, engaging, and focused session. Both students and instructors agreed they would
like to see the ARS format expanded across the curriculum and incorporated
into these sessions.
Table 1. In Class Student Survey Questions
Endocrinology
Concurrent Session
8:30 AM
Friday, January 28, 2011
61
AN UNUSUAL CASE OF SUPPURATIVE THYROIDITIS
CAUSED BY ESCHERICHIA COLI AND ENTEROCOCCUS
FAECALIS
Bouchonville M, Kapsner P University of New Mexico, Albuquerque, NM.
Case Report: Acute suppurative thyroiditis is an uncommon inflammatory
thyroid disease caused by bacterial infection. Affected individuals are typically susceptible to infection due to immunosuppression or anatomical abnormalities. The most commonly observed organisms in this condition are
Streptococcus and Staphylococcus species. Suppurative thyroiditis involving
Escherichia coli has been rarely reported in association with urinary tract
infections. We report an unusual case of suppurative thyroiditis caused by
Escherichia coli and Enterococcus faecalis in a patient with a dental abscess.
A 50 year-old woman presented to the ER with fevers, tender swelling of
her right neck, dysphagia, and dyspnea one week after being treated for a
dental abscess. WBC count was 12.9K (normal 4Y10K cells/mm3), TSH 1.02
(normal 0.4Y4.5 UIU/mL), total T4 16.3 (normal 4.7Y11.0 ug/dL), free T4 1.1
(normal 0.8Y1.6 ng/dL), total T3 251 (normal 84Y172 ng/dL), and thyroglobulin 5,831 (normal 1.6Y59.9 ng/mL). CT of the neck demonstrated a
large complex hypervascular thyroid nodule measuring 7cm in maximum
diameter with significant mass effect on the trachea, extension into the
superior mediastinum, and evidence suggesting neoplasm versus suppurative thyroiditis. Fine needle aspiration for cytology demonstrated a follicular
lesion, and culture of the aspirate was positive for Enterococcus faecalis and
E.coli. A barium swallow study demonstrated no evidence of pyriform sinus
fistula or brachial arch anomaly. Dental consultation was obtained for tooth
extraction, and the patient was treated with parenteral antibiotics with rapid
reduction in size of the neck mass. She subsequently was referred for hemithyroidectomy confirming the follicular lesion to be a benign adenoma.
The described case of acute suppurative thyroiditis caused by E.faecalis and
E.coli in a patient with dental abscess has, to the best of our knowledge,
never been reported and emphasizes the need to investigate the cause of
this potentially life-threatening condition. We review the relevant literature
regarding acute suppurative thyroiditis as well as a recently proposed algorithm for the management of this rare disorder.
62
DIAGNOSIS OF DIFFUSE IDIOPATHIC PULMONARY
NEUROENDOCRINE CELL HYPERPLASIA IN A PATIENT
WITH A COMPLEX PULMONARY HISTORY
Sadler C, Kapsner P Univ. of New Mexico, Albuquerque, NM.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Case Report: Diffuse idiopathic neuroendocrine cell hyperplasia (DIPNECH) is a rare syndrome with fewer than 50 cases reported. It presents with
cough, progressive dyspnea and irreversible airflow obstruction in middle
aged, non-smoking females. It results from hyperplasia of the pulmonary
neuroendocrine cells scattered through the bronchial tree and is likely part of
the spectrum of carcinoid tumorlet and tumor. We present a case of a patient
with a complicated pulmonary history diagnosed with DIPNECH.
A 56 year old female with a history of asthma, obesity and sleep apnea
presented with progressive dyspnea and hypoxemia. Initial concern for
worsening asthma, congestive heart failure and pulmonary embolism were
not substantiated. Patient had a history of pulmonary nodules noted in the
1990s, followed clinically. On computed tomography scan in 2008 these
nodules were noted to have grown leading to a left video-assisted thoracoscopic upper and lower lobe wedge resections and thoracotomy with mediastinal lymph node dissection. A diagnosis of two carcinoid tumors and
multiple meningothelial nodules was made. Follow up octreotide scan
showed no residual disease. However over the next year, patient_s dyspnea
and hypoxemia significantly progressed. Symptoms were initially felt to be
possibly related to meningotheliomatosis, and patient underwent a trial of
RU486, 200 mg/week. Patient did not show the expected clinical improvement. Serial monitoring of serum chromagranin A showed increasing levels
from 21 ng/ml post wedge resection up to 154 ng/ml (normal 0Y50 ng/ml).
Due to the patient_s complex pulmonary disease and progressive symptoms,
consultation was undertaken at National Jewish Health. Review of pathology
identified carcinoid tumorlets and Bproliferation of single neuroendocrine
cells confined to the bronchiolar epithelium.[ A diagnosis of DIPNECH
was made. Patient was subsequently treated with sandostatin LAR with
improvement in her chromagranin A levels and symptoms.
This case demonstrates a symptomatic presentation of the rare syndrome
DIPNECH in a patient with multiple underlying pulmonary pathologies. We
will review the literature relevant to the diagnosis and management of
DIPNECH.
63
ATYPICAL CASE OF THE MC CUNE ALBRIGHT
SYNDROME
Rastogi MV, Marks DL, Boston B Oregon Health and Science University,
Portland, OR.
Purpose of Study: The Mc Cune Albright syndrome is a clinical syndrome
that consists of a triad of precocious puberty, café au lait spots and fibrous
dysplasia. Its frequency ranges from one in 100, 000 to 1 in 1 million patients.
Precocious puberty is the most common presenting symptom especially in
girls occurring in up to 70Y90%. However, in males precocious puberty
presents in only 15%. In all known cases, the syndrome is due to a somatic
mutation in exon 8 or 9 of the alpha subunit of the stimulatory G protein.
This leads to constitutive activation as a result of impaired GTPase activity,
elevated adenyl cyclase activity and elevated cAMP levels. However, since
precocious puberty is rare in males, we proposed that the mutation in a male
with features of MAS and precocious puberty might be different than in
females. Our purpose was to characterize the defect leading to the Mc Cune
Albright syndrome in a young male with café au lait spots, fibrous dysplasia
and precocious puberty secondary to leydig cell tumors.
Methods Used: Human leydig cells were isolated from resected leydig cell
tumors. These cells were propagated and developed into a cell line. The Gs
alpha subunit was sequenced from paraffin embedded specimens. This sequencing was repeated on CDNA isolated from the patient_s leydig cells.
Cells were stimulated with HCG and analyzed for evidence of cAMP production and media was assayed for steroid production. An existing mouse
leydig cell line was used as a control.
Summary of Results: Sequencing of the entire Gs alpha gene using
genomic DNA from paraffin embedded specimens and CDNA isolated from
the leydig cell culture failed to detect any mutations including the usual
mutations in exon 8 or 9. The subject_s cultured leydig cells were analyzed for
cAMP production and found to have no production at baseline with a dose
dependent increase similar to mouse control. Media assayed for pregnenolone revealed elevated production at baseline with no increase with
HCG stimulation. In contrast, a mouse control showed no baseline steroid
production with an increase on HCG stimulation.
Conclusions: We present the first case of atypical Mc Cune Albright syndrome in which there is not a mutation in GS Alpha. This defect appears to be
in the pathway downstream from cAMP.
105
Western Regional Meeting Abstracts
Journal of Investigative Medicine
64
INFLUENCE OF ETHNICITHY IN SUPPRESSION AND
RECOVERY OF SPERMATOGENESIS IN MALE HORMONAL
CONTRACEPTIVE TRIALS
Ilani N1, Liu PY2, Swerdloff RR1, Wang C1 1Harbor-UCLA Medical Center
and Los Angeles Biomedical Research Institute, Torrance, CA and
2
Woolcock Institute of Medical Research, University of Sydney, Sydney,
NSW, Australia.
Purpose of Study: Unintended pregnancy remains an important global
problem. Male hormonal contraceptive studies in the last three decades
demonstrate efficacy and reversibility of this method in men. In these studies,
it was noted that ethnicity may influence the degree of sperm suppression and
recovery. For this reason, we conducted an integrated analysis of available
data from previous studies to examine the influence of ethnicity in male
hormonal contraception.
Methods Used: De-identified individual subject data were supplied by
investigators of 20 single-, and 10 multi-center studies spanning five continents (North and South America, Europe, Asia and Australia) and completed before 2007 through a standardized worksheet that was approved by
the Harbor-UCLA Medical Center Institutional Review Board. Data were
available from 1756 men and analyzed by Kaplan-Meier and Cox regression.
Summary of Results: Our integrated analysis shows: 1) Male hormonal
contraception using androgens alone suppress sperm output in up to 90% of
Asian men but only up to 80% in Caucasian men. 2) Addition of a progestin
causes suppression of sperm output to G1 million/ml in nearly all Asian
men and over 90% of Caucasian men. 3) All men will eventually recover
sperm output irrespective of ethnicity, although recovery is more rapid in
Asian men.
Conclusions: Ethnic differences in the suppression of spermatogenesis
suggest that testosterone alone male contraceptive methods may not be
suitable globally. Addition of a progestin quantitatively overcomes this ethnic
difference and results in adequate suppression of sperm concentrations and
probably contraceptive efficacy for all men.
65
ESTRADIOL17A (E2) ATTENUATES THE GNRH MEDIATED
INCREASE IN INTRACELLULAR CA2+
Mrdutt M1, Magee C2, Cantlon J2, Murtazina D2, Sanborn B2, Clay C2
1
Univ of Washington, Seattle, WA and 2Colorado State Univ., Fort Collins, CO.
Purpose of Study: Controlling the ovulatory LH surge underlies fertility
regulation. The LH surge is initiated by E2 actions at the hypothalamus and
the pituitary to increase GnRH receptor expression on gonadotropes - the LH
producing cells. Interestingly, however, E2 acutely but transiently suppresses
GnRH mediated LH release thus increasing pituitary content of LH just prior
to the surge. We hypothesized that E2 acutely attenuates the GnRH elicited
increase in intracellular Ca2+ - the cellular event underlying GnRH mediated
LH secretion.
Methods Used: We established cultures of pituitary cells from transgenic
mice in which expression of a fluorescent protein (YFP) is confined to
gonadotropes. The distinct excitation spectra of YFP and the Ca2+ indicator
FURA allowed for selection of gonadotropes and subsequent imaging of
GnRH induced changes in intracellular Ca2+. After 18 hr in culture, cells
were Bloaded[ with FURA for 1 hr. Cells were then incubated in Ca2+
containing or Ca2+ free media and received 100nM E2 or vehicle followed by
a 2nM GnRH pulse at 2 and 12 min post-E2. Increases in intracellular Ca2+
were determined by a change in FURA emission spectra. Both amplitude
(peak height) and magnitude (area under curve) of the Ca2+ response in 136
cells was analyzed 1 min post-GnRH.
Summary of Results: Approximately 93% of YFP positive cells displayed
a Ca2+ response following the initial GnRH pulse. Of these, 84% displayed a
secondary Ca2+ response; however, in both Ca2+ and Ca2+ free conditions,
the magnitude and amplitude of the secondary response was reduced
(pG0.05) as compared to the initial response. E2 did not alter the percentage
of cells responding to either GnRH pulse. However, in both Ca2+ and Ca2+
free conditions, E2 further reduced (pG0.05) the ratio of secondary versus
primary response magnitude by 25% and 31%, respectively.
Conclusions: Consistent with our hypothesis, E2 attenuates the GnRH induced Ca2+ response in gonadotropes - a potential explanation for the acute
block of GnRH mediated LH release by E2 prior to the ovulatory LH surge.
The rapidity of the inhibitory effect of E2 suggests a non-nuclear mechanism
106
&
Volume 59, Number 1, January 2011
of action. If correct, new generations of E2 agonists/antagonists may allow
for fertility regulation without the myriad physiological effects resulting from
nuclear actions of E2.
66
UNDERSTANDING THE ACTIVIN-INDUCED PATHWAY FOR
ENDODERM DIFFERENTIATION FROM EMBRYONIC
STEM CELLS
Sinha A1, Kartikasari A2, Bhushan A2 1David Geffen School of Medicine at
UCLA, Los Angeles, CA and 2University of California at Los Angeles, Los
Angeles, CA.
Purpose of Study: The efficiency of deriving functional A-cells from embryonic stem cells (ESCs) for diabetes treatment is currently low. The first step
in this process is to direct ESC differentiation into endoderm cells using
Activin, a growth factor. However, how Activin promotes endoderm specification is unknown. Our aim was to determine epigenetic changes that promote
mouse endoderm differentiation from mouse-ESCs upon Activin-induction
and to identify the key players through which Activin exerts its effects.
Methods Used: Chromatin immunoprecipitation (CHIP) was used to
identify two histone modifications: the permissive methylated lysine4 on
histone3 (H3K4me3) and the repressive methylated lysine27 on histone3
(H3K27me3). CHIP was also used to assess binding of Smad2/4, a transcription factor involved in Activin signaling, and Jmjd3, an H3K27 demethylase, to endoderm gene promoters before and after Activin-induced
differentiation. Loss of function assays were performed for Smad2/4 and
Jmjd3, using shRNA knockdown.
Summary of Results: Upon differentiation, the bivalent H3K4me3H3K27me3 marks present on endoderm gene promoters in ESCs, were
replaced by the H3K4me3 marks in endoderm, indicating loss of the repressive H3K27me3 marks. A significant increase in binding of both
Smad2/4 and Jmjd3 was noted on endoderm gene promoters upon differentiation. Knockdown of Jmjd3, decreased the efficiency of Activin-induced
differentiation by 3-fold. Knockdown of Smad2/4, abolished Jmjd3 binding
to the promoter of an essential endoderm gene, Sox17, and decreased Sox17
expression by 3.5-fold as measured by qrt-PCR.
Conclusions: Upon endoderm differentiation, Activin signaling via Smad2/4
directs Jmjd3 binding to the promoters of endoderm genes. The binding of
Jmjd3 in turn removes the repressive H3K27me3 marks, allowing the expression of endoderm genes. This study may help increase the efficiency of deriving
functional A-cells from ESCs.
67
DEVELOPMENT AND OPTIMIZATION OF MONOCLONAL
ANTIBODIES TO ZINC TRANSPORTER 8 FOR
IMMUNOHISTOCHEMISTRY
Gitomer S, Sarkar S, Walter J, Wenzlau J, Lee C, Hutton J University of
Colorado School of Medicine, Aurora, CO.
Purpose of Study: Type 1 diabetes is an autoimmune disease in which the
body_s own immune system attacks self-antigens in pancreatic islet cells,
leading to a decrease in production and export of insulin. In type 1 diabetes,
autoantibody markers serve as predictors of disease, measures of pancreatic A
cell damage and potential targets of therapeutics. Optimized monoclonal
autoantibodies can be used to purify proteins and to study protein characteristics, such as protein trafficking and topology, all of which will be
important for further characterization of ZnT8Va recognized auto-antigen in
type 1 diabetes. The goal of this project was to develop and optimize ZnT8
monoclonal antibodies for use in immunohistochemistry.
Methods Used: Hybridoma cultures were created to produce antibodies
against the recombinant C-terminal and N-terminal domains of ZnT8, and the
specificity of the antibodies was tested using radioimmunoprecipitation
assays (RIAs). The supernatants from the hybridomas were used as primary
antibodies for both fluorescence microscopy and DAB histology staining.
Summary of Results: Both RIA and immunohistochemical assays showed
that four of the hybridomasV17H2, 10D7, and 4D2Vproduced antibodies
specific for the N-terminal of ZnT8, but none produced antibodies specific for
the C-terminal domain. These four antibodies consistently co-localized with
insulin antibodies in pancreatic A cells, and were generally excluded from
areas where glucagon and somatastatin antibodies bound.
Conclusions: Overall, three new monoclonal antibodies specific for the
N-terminal of ZnT8 were developed and characterized for use in both
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
human and mouse tissue histology. While further characterization and
titration is necessary, the utility of each of these monoclonals is now
described.
68
MULTIPLE AUTOIMMUNE DISORDERS IN NEW ONSET
TYPE 1 DIABETES
Triolo TM, Yu L, Miao D, Scrimgeour LA, Klingensmith GJ, Eisenbarth GS,
Barker JM University of Colorado, Aurora, CO.
Purpose of Study: Children with type 1A diabetes (T1D) are at risk to
develop multiple autoimmune disorders including autoimmune thyroid disease (AIT), celiac disease (CD), and Addison_s disease (AD). Assays are
available for thyroid peroxidase autoantibodies (TPO) of AIT, tissue transglutaminase antibodies (TTG) of CD, and 21-hydroxylase antibodies (21OH)
of AD. We analyzed these non-islet, organ-specific antibodies at T1D diagnosis and determined the number of individuals diagnosed with multiple
autoimmune disorders within six months of T1D onset.
Methods Used: Patients (n=491) were consented and screened for the
presence of non-islet, organ-specific autoantibodies at T1D onset and followed for the development of AIT, CD, and AD. Eligibility included diagnosis of T1D, age 0 to 30 years, and T1D antibodies obtained within 3 months
of T1D diagnosis. HLA genotyping was performed on 457 subjects with
DNA available.
Summary of Results: Individuals with antibodies associated with AIT,
CD, and AD at T1D diagnosis were followed for progression to clinical
disease. Of the 487 with positive T1D antibodies, 160 (32.9%) had AIT, CD
or AD antibodies at T1D diagnosis. 25% (122/487) were positive for thyroid
peroxidase antibodies (TPO), 11.7% (57/487) were positive for tissue
transglutaminase antibodies (TTG) and 1.0% (1/487) were positive for 21hydroxylase antibodies (21OH). Throughout follow-up, 42 (8.6%) had evidence for progression to clinical disease (AIT, CD, or AD) and of these, 2
individuals had multiple autoimmune diseases in addition to T1D. DNA was
available for 93.4% of subjects. The high-risk diabetes related genotype HLA
DR3/4-DQ8 was present in 27.2% (31/114) of TPO positive (NS), 39.6%
(21/53) of TTG positive (NS), and 60% (3/5) of 21OH positive subjects (NS),
compared with 26.3% (80/304) of those with no additional autoantibodies.
Conclusions: In conclusion, one-third of patients newly diagnosed with
T1D are positive for at least one organ specific autoantibody. Within
6 months of T1D diagnosis 15.2% (28/184) of these were diagnosed with an
additional autoimmune disease. Presence of these other organ specific
autoantibodies is not related to the highest-risk T1D HLA genotype.
69
A CASE OF AN EXTREMELY AGGRESSIVE PITUITARY
TUMOR
Lovato C, Kapsner P, Garimella M Univ. of New Mexico, Albuquerque, NM.
Case Report: Pituitary tumors are relatively common tumors and account
for about 15% of intracranial neoplasms. The majority of pituitary tumors are
benign adenomas. About 0.2% of pituitary tumors are carcinomas. Pituitary
carcinomas pose a diagnostic challenge as they can have an initial presentation similar to invasive pituitary adenomas. However, these tumors typically
have multiple recurrences and eventually develop craniospinal and/or systemic metastases. We describe a case diagnosed as an aggressive pituitary
tumor which may in fact have been a pituitary carcinoma.
A 59 year old women presented multiple times over 4 weeks with
headaches, decreased visual acuity, diplopia, nausea, and vomiting. She was
initially diagnosed and treated for cluster headaches. Her symptoms failed to
improve and a few weeks later she developed ptosis of her right eye. An MRI
of the brain showed a large mass in the sellar region with invasion into the
clavus, sellar floor, anterior temporal lobes, and compression on the right
optic nerve. Prolactin, cortisol, ACTH, LH, FSH, and IGF-1 levels were
normal. The patient underwent debulking of the mass. Pathology was suggestive of an atypical invasive adenoma with a high Ki-67 proliferation
index and positive p53 immunoreactivity.
Three weeks after surgery, the patient developed recurrence of her
symptoms. Repeat MRI of the brain revealed the pituitary mass had doubled
in size with significant spread into the surrounding structures, including
compression of the optic chiasm. She underwent extensive debulking of the
tumor as well as radiation therapy. CT scan of the chest, abdomen, and pelvis
were negative for lymphadenopathy or other primary. Despite a second
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
debulking procedure and radiation therapy, the patient_s neurological deficits
progressed and repeat imaging of the brain a week later showed the tumor
increased in size as well as leptomeningeal spread to the cervical, thoracic,
and lumbar spine. The patient subsequently developed respiratory failure
from acute aspiration and given her poor prognosis was made comfort care.
This case illustrates the importance of recognizing the features of pituitary carcinomas and agressive adenomas as they can have a significant impact
on prognosis in these patients. We review the literature regarding the diagnosis, etiology, and therapy of these rare entities.
70
LOW VITAMIN D LEVELS IN NORTHERN AMERICAN
ADULTS WITH THE METABOLIC SYNDROME
Jialal G, Cook T, Siegel D, Jialal I, Devaraj S UC Davis, Sacramento, CA.
Purpose of Study: Vitamin D (VitD) deficiency is a worldwide problem. A
growing body of evidence suggests that 25-hydroxyvitamin D [25(OH)D], a
generally accepted indicator of vitamin D status, is inversely associated with
adiposity, glucose homeostasis, lipid profiles, and blood pressure along with
its classic role in calcium homeostasis and bone metabolism. Metabolic
syndrome (MetS), is a cluster of cardiometabolic disease risk factors, that is
now a global epidemic and predisposes to increased risks for diabetes and
cardiovascular diseases. Since there is sparse data on VitD status in MetS in
North America especially, the aim of our study was to examine vitamin D
[25(OH)D] concentrations among adults with MetS in Northern California
(sunny climate) , but without diabetes or cardiovascular disease.
Methods Used: VitD levels were assayed in MetS subjects (n=44) and
healthy controls (n=37), using the LIAISON-25OH Vitamin D TOTAL
Assay.
Summary of Results: 25OHD levels were significantly decreased in
MetS compared to controls (22 vs 28 ng/mL, pG0.02). Excluding AfricanAmericans, or using BMI/waist circumference as covariates, did not alter the
significant differences. There was no significant difference between number
of features of MetS and 25OHD levels. 67% of controls and 84% of MetS had
insufficient 25OHD levels (G30 ng/mL) (p=0.1). 8% of controls and 30% of
MetS were deficient in 25OHD (G20 ng/mL; p=0.02, C vs. MetS). There were
no significant differences between groups in blood sampling in winter and
summer months (Chi-square p=0.47). Furthermore, we examined creatinine,
calcium and phosphate, since these could regulate 25OHD levels, however,
these were similar between groups. There were no significant correlations
of 25OHD levels with the different features of MetS except fasting glucose
(r=-0.29, p=0.04) and HOMA (r=-0.34, p=0.04) or biomarkers of inflammation (CRP, leptin, adiponectin).
Conclusions: Adults with MetS, living in Northern California, display
significantly decreased vitamin D status compared to age and gender matched
controls and this cannot be explained by adiposity, sun exposure, renal disease. Decreased dietary intake cannot be ruled out since it was not recorded.
Since there was an increased correlation with HOMA, further studies of
vitamin D supplementation in these subjects on subsequent risk of diabetes,
will prove instructive.
71
THE EFFECT OF A ONE-HOUR AEROBIC RUN ON 25-OH
VITAMIN D LEVELS IN TRAINED AND UNTRAINED
PREMENOPAUSAL WOMEN
Schneider LE1, Prowse M2, Berk L2,3, Daher NS2, Haddock B4 1Loma
Linda University, Loma Linda, CA; 2Loma Linda University, Loma Linda,
CA; 3Loma Linda University, Loma Linda, CA and 4California State
University San Bernardino, San Bernardino, CA.
Purpose of Study: To compare the adequacy of dietary oral intake of
Vitamin D and blood serum levels of 25-hydroxyvitamin D {25(OH)D}
found in low versus high fitness premenopausal women at baseline and after
a one-hour aerobic exercise.
Methods Used: 21 Trained and 14 Untrained women, maximal oxygen
uptake (VO2 max) means 42.2T5.3mL/kg/min and 27.9T3.3mL/kg/min
respectively, performed a one-hour weight bearing exercise at an aerobic
intensity of 70Y80% of their maximum heart rate. Serum levels were determined at pre-, post- and 30 min post-exercise for: Ca2+, iPTH, 17A-Estradiol,
bone formation and resorption markers C-telopeptide of type I collagen
(CTx), N terminal propeptide of type I procollagen (P1NP) and 25(OH)D
and adjusted for plasma volume shifts.
107
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Both groups increased serum 25(OH)D levels with
Purpose of Study: Mucopolysaccharidosis type IIIB (MPS IIIB; Sanfilippo
exercise: Trained pre- to post- (p=.01) and Untrained pre- to post- (p=.02).
Also, the 25(OH)D levels in the Trained were significantly higher than the
Untrained at pre- (p=.01), post- (p=.01) and 30 min post-exercise (p=.02).
In both groups, Ca2+ and iPTH increased from pre-exercise to post exercise
and at 30 mins post-exercise returned to pre-exercise levels.
Conclusions: Levels of 25(OH)D were significantly higher in the Trained
versus the Untrained group at all time points (pG.05). In the Untrained
women 25(OH)D mean levels were G30 ng/mL; whereas in the Trained
women, mean levels were 9 36ng/mL at all time points. In evaluation of
dietary intake, there were no significant differences between groups in calcium intake from foods or vitamins, consumption of fruits and vegetables,
protein, caffeine or sodium intake (p9.05). Calcium and Vitamin D supplementation were similar in both groups. These results would support the
recommendation that Vitamin D supplementation may be beneficial for all
populations, including premenopausal women functioning at higher levels of
fitness and outdoor activity. However, fitness appears to play a key role in
providing higher 25(OH)D blood levels in premenopausal women.
B) is a lysosomal storage disorder characterized by deficiency of >-N-acetylglucosaminidase (NAGLU) causing defective degradation and accumulation
of heparan sulfate. Enzyme replacement therapy (ERT) has been used to treat
lysosomal storage diseases but has not been successful for MPS IIIB. In
particular, ERT for MPS IIIB has been limited by inadequate cellular uptake
of recombinantly produced human NAGLU enzyme. In this study we create,
characterize, and test a novel enzyme: NAGLU fused to the insulin-like
growth factor 2 (IGF-II) peptide (rhNAGLU-IGF-II). The IGF-II motif was
selected to enable recombinant NAGLU to enter cells via the insulin-like
growth factor 2 binding site found on the mannose 6-phosphate receptor for
high affinity delivery to lysosomes.
Methods Used: Following the molecular cloning of the recombinant
NAGLU-IGF-II construct, rhNAGLU-IGF-II was stably expressed in
Chinese hamster ovary cells in secreted form and purified. Activity assays,
biochemical analysis and substrate kinetics were performed. Cellular uptake
by human MPS IIIB fibroblasts and inhibition assays were followed by
confocal microscopy to evaluate the subcellular distribution of rhNAGLUIGF-II in the fibroblasts. Glycosaminoglycan (GAG) storage reduction
was measured.
Summary of Results: The enzyme activity towards the substrate 4-MUNG
was comparable to wild-type rhNAGLU, with peak activation near lysosomal
pH. MPS IIIB fibroblasts and brain-derived cell lines readily took up the
fusion enzyme via receptor-mediated endocytosis that was inhibited by
competitive IGF-II peptide. Co-localization of rhNAGLU-IGF-II enzyme
with lysotracker red in MPS IIIB cells via confocal microscopy confirmed
lysosomal targeting. GAG storage was also reduced up to 50% (pG0.001) in
MPS IIIB cells after treatment with rhNAGLU-IGF-II when measured by
radioactive incorporation assay.
Conclusions: Our data suggest that the fusion enzyme, rhNAGLU-IGF-II,
can enter human cells and localize in the lysosomes. Future studies in vivo are
needed to more fully assess the clinical potential of rhNAGLU-IGF-II as an
ERT for Sanfilippo B syndrome.
Genetics
Concurrent Session
8:30 AM
Friday, January 28, 2011
72
IMPROVED CLINICAL OUTCOMES OF CHILDREN
IDENTIFIED THROUGH NEWBORN SCREENING WITH
DEFECTS OF VITAMIN B12 METABOLISM
Merritt J, Hale S, Hahn S University of Washington, Seattle Children’s
Hospital, Seattle, WA.
Purpose of Study: Most newborn screening (NBS) programs in the United
States have now expanded their screens to include methylmalonic and propionic acidemia based on the propionyl-carnitine (C3) level. Although elevations of C3 are specific not only for these two primary diseases but also a
number of Bsecondary disorders[ of cobalamin and methionine metabolism.
Unfortunately there is only limited information available describing the effect
NBS has had upon the clinical disease and outcome for these secondary
disorders.
Methods Used: We identified a series of patients found to have cobalamin C
disease, the most common of the cobalamin metabolism disorders, in the first
year of life in Washington state.
Summary of Results: Six patients were identified: 3 from newborn
screening and 3 by onset of clinical disease. Average C3 and methionine
levels were 8.4 uM and 6.8 uM, respectively, for all patients. Three patients
were diagnosed with Cobalamin C disease after presenting with clinical
symptoms including failure to thrive, developmental delay, seizures, and
hypotonia. All patients were started on treatment with folinic acid, betaine,
L-carnitine, and intramuscular hydroxocobalamin injections. All affected
patients have subsequently shown clinical improvement in growth and
development. Those infants identified presymptomatically through NBS have
presented with nystagmus or other mild neurodevelopmental delay. All
patients have demonstrated speech delay despite treatment. Those symptomatic patients have shown improvements in seizure severity, gross motor
development, and weight gain. All patients have had dramatic improvements
in all biochemical markers and have maintained mildly elevated total
homocysteine (range 20Y50 uM) and methionine levels in the normal range.
Conclusions: Our clinical observations support previous findings that
newborns who receive early treatment due to newborn screening have
improved outcomes than those who do not receive treatment until they have
developed more substantial clinical disease. This provides further evidence
of additional benefits from expanded newborn screening and that further
attention should be given to patients with low methionine values on the
newborn screen so that treatment may be initiated as soon as possible.
73
INSULIN-LIKE GROWTH FACTOR II PEPTIDE FUSION
ENABLES UPTAKE OFALPHA-N-ACETYLGLUCOSAMINIDASE
INTO MUCOPOLYSACCHARIDOSIS TYPE III B FIBROBLASTS
Sinow C, Kan S, Tippin B, Troitskaya L, DiStefano A, Todd A, Le S,
Dickson P LA BioMed at Harbor-UCLA, Torrance, CA.
108
74
BRAIN MRI/MRS FINDINGS IN PATIENTS WITH
SMITH-LEMLI-OPITZ SYNDROME (SLOS) AND
CORRELATIONS WITH CLINICAL AND BIOCHEMICAL
PARAMETERS
Flory G1, Merkens LS2, Merkens MJ2, Woodward W1, Kroenke CD1,
Roullet J2, Steiner RD2 1OHSU, Portland, OR and 2OHSU, Portland, OR.
Purpose of Study: SLOS is caused by inactivating mutations of the gene
coding for 7-dehydrocholesterol reductase (DHCR7), an enzyme that is
necessary for the biosynthesis of cholesterol. The purpose of the current
study is to identify brain characteristics in SLOS that correlate with disease
severity and change with interventions. If successfully identified, such
characteristics are likely to be useful both as diagnostic or prognostic tools as
well as treatment outcome measures for clinical trials.
Methods Used: Ten patients with a diagnosis of SLOS, ranging in age from
0.4Y18.2 years, served as subjects. An overall measure of disease severity
was obtained using a standardized battery of physical and neurocognitive
tests. Plasma levels of cholesterol and 7-DHC (the cholesterol precursor that
accumulates pathologically in SLOS), were measured. Using a Siemens 3T
TIM Trio MRI system, imaging and spectroscopic (MRI/MRS) data were
obtained under general anesthesia. The images were used to assess gross
structural abnormalities and the spectra to quantify brain lipid MRS intensity.
Summary of Results: Disease severity scores ranged from 0Y40. Plasma
cholesterol and 7-DHC concentrations ranged from 8Y131 and 0.2Y18 mg/dL,
respectively. Negative correlations were found between plasma cholesterol
levels and the overall clinical assessment of disease severity (r=j0.632,
p=0.025), gross structural abnormalities of the brain (r=j0.813, p=0.004),
and the size of the brain_s lateral ventricular system (r=j0.748, p=0.006).
In addition, a positive correlation was found between plasma 7-DHC levels
and brain lipid MRS intensity (r=0.738, p=0.012).
Conclusions: The data show that clinical and biochemical indices of SLOS
disease severity correlate with brain abnormalities. The finding of a positive
correlation between 7-DHC and the MRS measure of brain lipid intensity
corroborates an earlier anecdotal report of abnormally large brain MRS lipid
peaks in patients with SLOS - a measure that was reported to decrease
following cholesterol supplementation therapy. This possibility that brain
lipid MRS signal intensity may have diagnostic and prognostic value in
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
SLOS warrants further study as it may prove useful as an outcome measure
and biomarker in clinical trials.
75
46,XY DISORDER OF SEX DEVELOPMENT: REPORT OF A
FAMILY WITH AN APPARENTLY NOVEL X-LINKED FORM
Myers A, Bernstein J, Lopez G, Hudgins L Stanford University, Stanford, CA.
Case Report: Disorders of sexual development (DSD) are defined by discrepancy between external genitalia, gonadal and/or chromosomal sex.
Various forms have been reported to be inherited in autosomal dominant,
recessive, and more rarely, X-linked fashion. We report a family in which two
genetically male, maternal cousins presented with ambiguous genitalia
associated with impaired testicular function. Pedigree analysis is consistent
with X-linked inheritance and known forms of X-linked DSD have been
ruled out. The proband presented at birth with ambiguous genitalia characterized by microphallus, bilateral undescended testes and a pigmented midline raphe extending posteriorly over fused scrotal/labial structures. Small
mullerian duct remnants were seen on cystoscopy. Endocrine workup in the
neonatal period included a low testosterone level and undetectable antimullerian hormone, implying a Sertoli cell dysfunction or anorchia. After 3
doses of hCG, testosterone level did not increase. At orchiopexy, a small
testis was noted on the right and only remnant tissue was found on the left.
Based on the physical and laboratory findings, endocrinology suggested that
this child_s condition was most consistent with Bvanishing testes[. The
proband_s mother_s sister_s fetus was noted to have discrepant karyotype
(46,XY) and genitalia (female appearing) on prenatal ultrasound. Pregnancy
was terminated at 22 weeks. Ambiguous genitalia was noted on external
examination of the fetus. 105k oligonucleotide array CGH, performed on
fetal tissue to rule out NR0B1 duplication, was normal. This apparently
X-linked form of 46,XY DSD is not consistent with any of the three well
described causes of X-linked DSD: alpha thalassemia/X-linked mental
retardation, androgen insensitivity syndrome, and duplication of NR0B1
formerly known as DAX1 which is associated with congenital adrenal hypoplasia. This appears to be a novel, X-linked form of DSD with two 46, XY
maternal cousins presenting with ambiguous genitalia associated with impaired testicular development. Genetic studies of the family are underway to
identify a locus/loci that may be associated with the familial disorder. The
underlying genetic etiology for the findings in this family may shed light on
one of the causes for Bvanishing testes[.
76
XP11.4 DUPLICATION IN A SIMILARLY AFFECTED
MOTHER AND SON: A NEW RECOGNIZABLE SYNDROME
WITH EXPRESSION IN THE FEMALE
Hall B University of Kentucky, Lexington, KY.
Purpose of Study: Identify phenotype of Xp11.4 duplication. Generally,
Xp duplications of an extensive degree such as Xp11-Xp23 have not resulted
in clinical abnormalities in females. Only 3 cases in females involving Xp11Xp14 duplication have resulted in varied clinical abnormalities plus developmental delay and speech difficulties. This was explained on skewed X
inactivation, both random and preferential. Could smaller Xp duplications
be the cause of recognizable phenotype? A mother/son pair offered proof that
this was so.
Methods Used: Phenotype analysis of mother and son.
A 6 year-old boy was seen for developmental delay, dysmorphic facial
features, speech difficulties, and behavior problems. He was the product of
a normal pregnancy and he had normal prenatal and postnatal growth (ht.,
wt., h.c.). He had been healthy except for dislocation of the same elbow on
3 occasions. His mother had speech problems, particularly in expressive
language, and she was manic depressive, but of normal intellect. Both the
son and his mother had similar facial features which included triangular face,
broad forehead, blue sclera, malar hypoplasia, prominent nasal tip, narrow
palate, and smooth skin. Two differences were the mother had micrognathia
and the son had pectus excavatum and bicuspid aortic valve and mildly
dilated aortic annulus and root. An X-linked disorder was considered because
the mother was somewhat milder, but no phenotype match was found.
Summary of Results: Both mother and son had Xp11.4 duplication as an
explanation for their physical features including speech difficulties and
possible psychiatric/mental problems.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
After 4 years of follow-up a CGH ARRAY was performed and it showed a
duplication at Xp11.4 which was 0.07Mb to a maximum of 0.131 Mb in size.
The mother showed the same duplication and is presently being studied for
skewed X inactivation. The Xp11.4 duplication does duplicate the TSPAN7
gene which is known to cause non-syndromic mental retardation.
Conclusions: Phenotype of Xp11.4 duplication results in a recognizable
phenotype with variable effect on mentation plus expressive language difficulties, behavioral/psychiatric problems, and possible connective tissue
abnormalities.
77
OSTEOGENESIS IMPERFECTA AND GENERALIZED
ARTERIAL CALCIFICATIONS OF INFANCY
Monteil DC, Kunz GJ, Maurer ST, Willis MJ Navy Medical Center San
Diego, San Diego, CA.
Case Report: The purpose of this abstract is to describe an infant afflicted
with two unrelated genetic disorders, Osteogenesis Imperfecta and Generalized Arterial Calcification of Infancy. Osteogenesis Imperfecta, OI, is an
autosomal dominant disorder of type 1 collagen causing bone fragility, short
stature, blue sclera, scoliosis, progressive long bone deformity, hearing loss
and dentinogenesis imperfecta. For OI type II, the most severe form, most
infants die in the perinatal period from respiratory insufficiency. Generalized
Arterial Calcification of Infancy, GACI, is a rare fatal autosomal recessive
disorder thought to be caused by decreased inorganic phosphate production
leading to increased hydroxyapatite deposition in blood vessel walls. This in
turn leads to progressive arterial stenosis and decreased elasticity in vessel
walls. Most infants affected with GACI die within the first year of life due to
progressive stenosis of the coronary arteries resulting in myocardial infarction. Interestingly both these disorders have been treated with bisphosphonates with varying success as reported in the medical literature.
The drug_s presumed mechanism of action in the two disorders differs.
Bisphosphonates are synthetic analogues of pyrophosphate and pyrophosphate is both an inhibitor of bone readsorption and an inhibitor of
hydroxyapatite deposition. We present a male infant diagnosed with OI type
II/III based on clinical evaluation, and confirmed with mutation analysis
(splice site mutation COL2A1 IVS37+1G9A) and collagen analysis in
fibroblasts. Echocardiography during the first days of life showed circumferential calcification of the pulmonary artery root. Repeat echocardiography
at 11 weeks of age showed progression of calcifications in the pulmonary
artery with the development of moderate pulmonary stenosis. Gated CT scan
demonstrated calcifications in other large and small arteries most consistent
with a diagnosis of GACI. Treatment with bisphosphonates was initiated and
patient is now 4 months of age. Both of these conditions are very frequently
fatal in the neonatal period. It is interesting to speculate whether the mechanism of disease in either of these disorders may alter the clinical course or
potentially even palliate the other.
78
EHLERS-DANLOS TYPE VIII, PERIODONTITIS-TYPE:
FURTHER DELINEATION OF THE SYNDROME IN A
4-GENERATION PEDIGREE
Reinstein E1, Wang RY2, Rimoin DL1, Wilcox WR1 1Cedars Sinai
Medical Center, Los Angeles, CA and 2Children_s Hospital Orange
County, Orange, CA.
Case Report: The Ehlers-Danlos Syndromes (EDS) are a family of inherited disorders of connective tissue that share the common feature of joint
hypermobility and skin abnormalities, significantly impacting the health and
well-being of the affected individual. To date, at least six subtypes with
distinct phenotypic features have been delineated; these include classic,
hypermobility, vascular, kyphoscoliotic, dermatosparaxis, and arthrochalasia
types. A few other rare subtypes have been described, including periodontitis,
valvular, and progeroid types. Due to the rarity of those entities, at present, it
is not clear whether they exist as autonomous forms of EDS. The periodontitis type of Ehlers-Danlos syndrome (EDS VIII - OMIM: 130080), was
first described by McKusick in 1972 and recognized as a distinct EDS subtype by Stewart et al in 1977. EDS type VIII is distinguished from other
subtypes by severe gingival recession and periodontitis leading to premature
loss of permanent teeth and resorption of alveolar bone by the second to third
decades of life, as well as lack of vascular and organ rupture. A limited
number of patients and pedigrees with this condition have been described. We
109
Western Regional Meeting Abstracts
Journal of Investigative Medicine
report a 4-generation EDS VIII kindred that has been followed over 12 years,
whose phenotype is generally characterized by joint hypermobility, normal
scar formation but eventual scar atrophy, and severe periodontal disease.
Descriptive statistics is presented for each clinical manifestation including
dental, musculoskeletal, dermatologic, facial features and other evidence of
tissue fragility. Similar to other subtypes of EDS, the age of onset and severity
of symptoms was variable amongst affected individuals. Our initial studies on
this pedigree do not show linkage to the previously reported EDS VIII locus
at 12p13, consistent with previously reported genetic locus heterogeneity.
79
COMPARTMENT SYNDROME, A RARE COMPLICATION
OF EHLERS-DANLOS SYNDROME TYPE IV
Sun A, Reinstein E, Rimoin DL Cedars-Sinai Medical Center, Los
Angeles, CA.
Purpose of Study: Ehlers Danlos Syndrome (EDS) type IV, also known as
vascular type, is caused by mutations in COL3A1 and is characterized by
extreme tissue fragility. Affected individuals display thin, translucent skin,
easy bruising, and develop severe aneurysms. Vascular rupture or dissection
and organ rupture are the presenting signs in 70% of adults, and the median
age of death is 48 years. Compartment syndrome as a complication of EDS
type IV is rare. We present a case of a 37 year old female with EDS type IV
who developed numerous episodes of compartment syndrome as a result of
aneurysms or vascular rupture following minor trauma. She had 25 episodes
involving various extremities, six of which had a documented elevation in
compartment pressure. She underwent 42 surgeries to treat the compartment
syndrome, including fasciotomies. A literature search revealed only three
other cases of compartment syndrome associated with EDS type IV. We
compare our case to those previously published and examine their varying
presentations, area of involvement, treatment modality, clinical course, and
other associated clinical features.
Methods Used: Retrospective chart review and literature search.
Summary of Results: Patient age ranged from 27Y47 years. There were
2 males and 2 females. Areas of involvement included the arm, leg, hand
foot, abdomen, and gluteus. Ruptured arteries were the posterior tibial,
gluteal, and bilateral renal. Documented compartment pressure ranged from
15Y80 mmHg. All patients were treated with fasciotomies except the individual with abdominal compartment syndrome who underwent abdominal
decompression. One patient also had arterial embolization prior to fasciotomy. Three patients had long term follow-up and were still living more
than 5 years after their episode. Only our index case had recurrent episodes;
the others were isolated incidents.
Conclusions: Compartment syndrome as a result of aneurysmal rupture is a
rare but serious complication of EDS type IV. Prompt recognition and
treatment are critical. Recurrence of compartment syndrome appears to be
uncommon and was seen in only 1 of 4 cases in this series.
80
DNA CpG DEMETHYLATION AT IGF-1 INTRON 2 STAT5B
ENHANCER SITE OCCURS AT ONSET OF ADOLESCENCE
Wiedmeier J, Liu S, Yu X, Callaway C, McKnight R, Lane R University of
Utah, Salt Lake City, UT.
Purpose of Study: Insulin growth factor 1 (IGF-1) mediates neurodevelopment, growth, and insulin sensitivity. Hepatic production of IGF-1 determines serum levels. Starting in adolescence, growth hormone (GH) regulates
IGF-1 expression through multiple STAT5b enhancer sites within the IGF-1
gene. The STAT5b enhancer within intron 2 (STEin2) appears to be particularly important. In the newborn mouse, STEin2 is flanked by DNA CpG
methylation. In the adolescent mouse, STEin2 undergoes significant histone
covalent modifications to allow GH signaling. However, little information
exists about if and when STEin2 undergoes DNA demethylation. This
information is important because 1) histone modification changes occur in
conjunction with changes in DNA methylation; and 2) diseases such as
intrauterine growth restriction affect hepatic IGF-1 expression. Characterizing the normal epigenetic regulation of GH-mediated IGF-1 will contribute
to determining how disregulation can lead to disease. We therefore hypothesize that DNA CpG demethylation occurs within STEin2 occurs during
adolescence.
Methods Used: Livers from C57B6 and B6:D2 mice were collected and
flash frozen at days 7, 21, 28, 35 and 90. Four CpG sites on both strands
110
&
Volume 59, Number 1, January 2011
around the IGF-1 intron 2 STAT5b enhancer were analyzed by bisulfite
sequencing.
Summary of Results: Overall, the IGF-1 STAT5b intro 2 enhancer was
hypermethylated (80% meCpG) at day 7 and relatively hypomethylated (25%
meCpG) by day 90. A dramatic decrease of DNA methylation occurred from
day 21 (36% meCpG) through day 35 (15% meCpG). Percentage of methylation on sense and antisense strands in both mouse strains were equivalent.
Conclusions: We conclude that the majority of DNA CpG demethylation
occurs in a relatively brief 2 week period between day 21 and 35 of life. This
period provides a window of study on which to focus upon the chromatin
modifying complexes that regulate IGF-1 expression in normal individuals,
as well as determine which of these complexes (or their components) are
disrupted by intrauterine growth restriction.
81
DUPLICATE GENETIC TESTING IN AN INTEGRATED
HEALTHCARE SYSTEM
Williams MS1, Collingridge DS2, Wood GM1, Gudgeon J1 1Intermountain
Healthcare, Salt Lake City, UT and 2Intermountain Healthcare, Salt Lake
City, UT.
Purpose of Study: Genetic testing for germline mutations is unique in that
a test only needs to be done once in a patient_s life. The purpose of this study
was to determine if two, commonly-ordered genetic tests were being performed more than once in our system.
Methods Used: The tests used for the analysis were Factor V Leiden (FVL)
and the Prothrombin Variant 20210A (PTM). Six years of data were extracted
from the electronic data warehouse (EDW). Duplicate tests were defined as
the specific test code being present 2 or more times in the unique patient
record separated by an interval of time. Data were also collected about the
facility where testing was done, the location within the facility, the name and
specialty of the ordering physician(s). Root cause and a variety of statistical
analyses were performed to identify patterns as well as system factors leading
to duplicate testing.
Summary of Results: Over the six-year period 4% of the FVL and 3.4% of
the PTM tests were duplicated. This resulted in nearly $50,000 (using Utah
Medicare rates) of waste for these tests over this period. Surprisingly, preliminary analysis showed that 20% of the duplicate tests occurred within one
week of the initial test. This suggests that communication in care transitions
(e.g. inpatient to outpatient or ICU to inpatient ward) may play a significant
role in this problem. Additional analyses are currently underway to better
characterize the predictive factors. Based on this information, root cause
analysis will be applied to identify the system failure that results in duplicate
testing.
Conclusions: Duplicate genetic testing is a problem in our system. Elimination of this duplication will save money with no negative impact on patient
care. Persistence of DT suggests there is a need for educating some providers
about the appropriate use of genetic testing for germline mutations as well as
exploring system changes that would prevent DT ordering.
82
DE BARSY SYNDROME; REPORT OF A PREVIOUSLY
UN-DESCRIBED CLINICAL FEATURE THAT SUGGESTS
NEW DIRECTION FOR ETIOLOGIC INVESTIGATION
Willis MJ 1Naval Medical Center San Diego, San Diego, CA and
2
Uniformed Services University, Bethesda, MD.
Purpose of Study: Explore the etiology of de Barsy Syndrome.
Methods Used: Literature review and clinical evaluation.
Summary of Results: De Barsy syndrome is a rare autosomal recessive
progeroid syndrome of unknown etiology. Consistent features in the fewer
than 30 reported patients include; pre and postnatal growth retardation,
progeriod appearance from birth, large fontanelles, large ears, clinched
hands, congenital hip dislocation, hyperextensible joints, severe mental
impairment, seizures and cloudy corneas. Additional connective tissue
abnormalities have been reported less frequently including scoliosis, easy
bruising, thin translucent skin, and other joint dislocations. In a number of
reports authors have identified tissue abnormalities in biopsies from individual cases including abnormalities of both elastic fibers and collagen.
Despite these studies, the genetic etiology of this disorder remains unknown.
Here we report a patient with phenotype of de Barsy syndrome including all
of the above features, who also has the previously unreported abnormality of
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
tortuous cranial and retinal arteries. Systemic arterial tortuosity is a feature of
a number of connective tissue disorders due to mutations in both genes for
structural proteins such as collagen and involving TGFbeta signaling. Cranial
and retinal arterial tortuosity has also been described with Menkes disease, a
disorder of copper transport leading to generalized copper deficiency. Copper
dependent enzymes are important for lysine-derived cross-links in both
elastin and collagen. Not unexpectedly therefore, Menkes patients have laxity
of joints and skin in addition to the more debilitating features of severe mental
retardation and seizures.
Conclusions: Tortuous cranial arteries in this patient with de Barsy syndrome is an addition feature of connective tissue dysfunction. The commonality of this feature with Menkes disease could suggest new direction of
investigation as to the etiology of de Barsy syndrome.
83
ELUCIDATING THE COMPLEXITY OF EPILEPSY: CLUES
FROM 2Q23.1 DELETION
Noh GJ, Graham Jr JM Cedars Sinai Medical Center, Los Angeles, CA.
Case Report: Epilepsy is one of the most common and genetically intricate
neurological disorders in children. Previous studies have helped elucidate
genes, which impart susceptibility to epilepsy. Since the introduction of the
array CGH, additional deletion and duplication syndromes have been delineated. Current research suggests an estimated 10Y30% background risk
for copy number variations in patients with seizures. We report two patients
who presented with seizures, developmental delay and dysmorphic features
due to chromosomal deletions at 2q23.1. Case 1 is a 3 year old male with
seizure disorder, deafness, microcephaly and dysmorphic features: microcornea, curly eyelashes, bushy eyebrows, hypoplastic nose, low-set ears with
an over folded anterior helix, prominent hallucal creases, single transverse
palmar creases, absent patella and severe neurological impairment. Cytogenetic analysis revealed a deleted region of 2q23 to 2q31.1. Case 2 is a
2 year old female with developmental delay, dysmorphic features, hypotonia
who presented with new onset seizures. Dysmorphic findings included
Darwinian tubercle, hypotonic facial features with short upturned nose, and
thickened tongue. An oligonucleotide microarray showed a 0.3 Mb deletion
involving 6 oligonucleotides within 2q23.1. Common physical features included bifrontal narrowing and low anterior hairline, short neck with redundant neck skin, fifth digit clinodactyly, with generalized brachydactyly.
In a literature review of 2q23.1 deletion, the critical region that has been
implicated as a seizure gene is MBD5, which seems to be responsible for
regulating DNA methylation in central nervous system tissue. Our first
patient_s larger area of deletion also involves the SCN gene cluster and
DLX1/2 genes. Abnormal SCN1A channels affect GABA neurons and the
associated seizures respond optimally to antiepileptic drugs that bind to the
GABA receptor. The DLX genes play a major role in controlling craniofacial
patterning and differentiation and survival of forebrain inhibitory neurons,
suggesting that haploinsufficiency might indeed be responsible for our
patient_s phenotype (i.e. seizures, micropthalmia). Chromosomal microarray
serves not only as a diagnostic tool, but also as a window into the pathophysiology and treatment of genetic encephalopathies.
84
A PRENATAL DILEMMA; ELLIS-VAN CREVELD AND
NEUROFIBROMATOSIS
Jelin AC, Perry H, MacLean J, Rauen K UCSF, San Francisco, CA.
Case Report: We report a case of a couple who presented for prenatal
counseling because of a maternal history of café-au-lait macules, axillary and
inguinal freckling and multiple cutaneous neurofibromas. On exam, findings
were consistent with a clinical diagnosis of Neurofibromatosis Type 1 (NF1).
Surprisingly, her partner was found to have a phenotype that consisted of
dwarfism, dolicocephaly, multiple lower lip frenula, a narrow trunk, short
limbs, and postaxial polydactyly. He also had a history of a partial AV canal
with an ostium primum, atrial septal defect and Eisenmenger_s physiology.
His presentation was concerning for Ellis-van Creveld (EVC). Neither had
previously undergone molecular genetic testing. Originally the couple was
mainly concerned about a possible diagnosis of NF1 for their unborn child.
After examination and counseling the risk of EVC was also presented to the
family who then underwent molecular genetic testing.
The male patient was found to be homozygous for a c.1238delA mutation
of the EVC2 gene consistent with Ellis-van Creveld. The female patient
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
was heterozygous for a truncating mutation (c.3567delA) in the NF1 gene.
The female then elected to also have testing for EVC carrier status. An
amniocentesis was performed and a targeted mutation analysis for the disease causing NF1 gene was requested.
Elis-van Creveld (EVC) syndrome is an autosomal recessive skeletal
dysplasia resulting from a mutation in either the ECV or EVC2 gene. The
exact protein responsible for the phenotype has yet to be characterized.
Patients have disproportionate dwarfism with short limbs, short ribs, thoracic
dysplasia and postaxial polydactyly. Cardiac defects occur in 60% of patients
and are usually in the form of an artio-ventricular septation or a single atrium.
This case highlights the importance of joint couple counseling in prenatal
cases. Although this couple originally presented for counseling in regards to a
diagnosis of NF1, the paternal exam was concerning for another syndrome
for which we were also able to offer molecular genetic testing. Although
characteristics of EVC may be identified on prenatal ultrasound, the ability
to perform a molecular diagnosis provides a definitive result to concerned
patients.
Health Care Research I
Concurrent Session
8:30 AM
Friday, January 28, 2011
85
BONE SCAN OVERUSE IN THE STAGING OF
PROSTATE CANCER
Palvolgyi R 1, Daskivich T 1, Kwan L 1,3, Chamie K 1,3, Litwin M 1,2,3 1UCLA
David Geffen School of Medicine, Los Angeles, CA; 2UCLA School of
Public Health, Los Angeles, CA and 3UCLA, Los Angeles, CA.
Purpose of Study: With contemporary widespread PSA screening, more
patients exhibit low- and intermediate-risk prostate cancer, thus reducing the
need and yield of bone scan imaging. We aimed to determine the utilization
and subsequent positivity rates of bone scan imaging in a contemporary
Veterans Affairs (VA) cohort of men with prostate cancer.
Methods Used: We retrospectively reviewed the charts of 1597 men
diagnosed with prostate cancer between 1997 and 2004 at the Greater Los
Angeles and Long Beach VA Medical Centers. We used univariate analysis to
measure association between patient (age, race, comorbidity) and tumor
(PSA, clinical stage, Gleason score) characteristics with bone scan use and
positivity. We conducted the analysis for scans that were and were not
clinically indicated, according to established AUA guidelines.
Summary of Results: Out of 1121 men not indicated for nuclear imaging,
452 (40%) received a bone scan with only 6 (1%) returning positive. Among
the same group, increasing PSA, clinical stage, Gleason score, and subsequent D_Amico tumor risk, were all positively associated with bone scan
overuse, but not with corresponding positivity rates. Univariate analysis
showed age, PSA, clinical stage, Gleason score, and treatment type as significant predictors of bone scan overuse.
Conclusions: Bone scan overutilization in men with clinically localized
prostate cancer results in unnecessary patient anxiety, time consumption, and
significant economic waste for the VA. If the 40% overutilization rate for
clinically localized prostate cancer reflects national overuse patterns, then
implementing a quality improvement initiative to minimize overutilization
would translate an annual savings of $9,409,680 for the VA, without compromising oncologic outcomes.
111
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
86
SCREENING FOR LYNCH SYNDROME USING COLORECTAL
CANCER IMMUNOHISTOCHEMISTRY
88
INSURANCE STATUS AND EMERGENCY DEPARTMENT
UTILIZATION IN PATIENTS WITH ACUTE STONE EPISODES
Khaki A, Madlensky L University of California, San Diego School of
Medicine, La Jolla, CA.
Purpose of Study: Current guidelines recommend that colorectal cancer
(CRC) patients with abnormal immunohistochemical (IHC) staining for
mismatch repair proteins be referred for genetic consultation to determine if
germline genetic testing for Lynch Syndrome is appropriate. Our objective
was to summarize patterns of IHC among CRC patients seen at a single
institution over a 6 year period.
Methods Used: Chart reviews were conducted on all CRC adenocarcinoma
cases at UCSD Medical Center from 2004Y2009. Covariates included age
and sex of patient, family history of CRC, type of insurance, year of diagnosis, tumor location and other histopathologic features.
Summary of Results: A total of 651 patients were identified in the hospital
tumor registry as having a CRC diagnosis in the study time period, of whom
331 had their tumor reviewed by pathology. Of these, 154 (46.5%) had IHC
completed, with younger patients, those with a relative with CRC, and those
with Medicare or private insurance more likely to have IHC completed. Of
those with completed IHC, patients with right-sided tumors or positive CRC
family history were more likely to have abnormal staining. Patients with
abnormal staining were twice as likely to be referred for a genetics consultation than those with normal IHC (42.4% vs 20.3%, p= 0.009).
Conclusions: Patients who are more likely to have Lynch Syndrome can be
efficiently identified through IHC and appropriately referred for a genetics
consultation. In addition, some patients in the study with abnormal IHC did
not have typical risk factors, and so systematic IHC for all patients may
be appropriate.
Han DS1, Schroeder G1, Richards G2, Tenggardjaja C2, Bowman R2,
Agarwal G2, Baldwin D2 1Loma Linda University, Loma Linda, CA and
2
Loma Linda University Medical Center, Loma Linda, CA.
Purpose of Study: According to the U.S. Census Bureau, 50 million people
in the United States are uninsured. It has been suggested that those who are
uninsured follow-up in the Emergency Department (ED) at a higher
frequency due to inability to access specialized care. The primary objective
of this study is to examine the relationship between insurance status and
ED usage among patients during an acute stone episode and evaluate their
follow-up care.
Methods Used: This is a single-center, IRB approved, retrospective study
from January 2007 to July 2009. Patients presenting to the ED with possible
acute stone episodes were identified by ICD-9 codes for nephrolithiasis,
ureterolithiasis, renal colic, and abdominal pain. Patients were excluded if
they did not have radiographic data confirming the presence of upper urinary
calculi. Follow-up location, number of stones, and number of ED follow-up
visits for the same stone episode were collected by chart review and telephone
contact and stratified by insurance status. Data were analyzed using two-tailed
Fisher_s exact and independent samples t-Test with a pG0.05 considered
significant.
Summary of Results: Six hundred and 10 patients were identified. Of these
321 patients were identified that met the criteria for an acute stone episode. In
136 of these patients follow-up location could not be identified. One hundred
and 85 patients were included for analysis based on nephrolithiasis or ureterolithiasis seen on CT and a chief complaint of abdominal or flank pain. Of
the 136 patients with unknown follow-up, 103 were insured (38% of total
insured) compared to 33 who were uninsured (66% of total uninsured),
pG0.001. In total, 143 of the insured patient population (85%) had at least one
follow-up visit (ED, primary care, and/or urology) after initial presentation
for an acute stone episode. In contrast, only 9 of the uninsured population
(53%) had a follow up visit, p=0.003.
Conclusions: Those without insurance utilize the ED at a higher frequency
for follow-up of acute stone episodes. Furthermore, those without insurance
are less likely to follow-up with a urologist after an acute stone episode.
Knowledge of follow-up patterns can guide health care systems to maximize
patient care.
87
CLINICAL AND ECONOMIC IMPACT OF FORMULARY
RESTRICTION ON PHACOEMULSIFICATION
Burns JD1, Tsai PL2 1University of Arizona College of Medicine, Tucson,
AZ and 2University of Arizona College of Medicine, Tucson, AZ.
Purpose of Study: Our purpose was to examine the effect of formulary
restriction on clinical outcomes and perioperative costs in phacoemulsification (phaco).
Methods Used: Retrospective analysis of 79 eyes that underwent phaco to
determine whether subjects_ use of non-preferred fluoroquinolones (ofloxacin
and moxifloxacin) or NSAIDs (bromfenac and diclofenac) due to formulary
restriction over preferred agents (gatifloxacin and ketorolac) had an impact on
clinical outcomes or personal costs. The change in visual acuity (VA), the
absolute value of difference between post-operative and expected spherical
equivalent (abs MRx diff), change in intraocular pressure (IOP), rate of
endophthalmitis or other serious adverse events, and the difference in the
subjects_ cost were compared between the groups.
Summary of Results: Subjects who received ofloxacin (n=34, j0.70,
p=0.008) and moxifloxacin (n=6, j0.54, p=0.034) had greater improvements
in LogMAR VA over the gatifloxacin group (n=39, j0.32); however, they
began with significantly worse vision than the gatifloxacin group [ofloxacin:
(0.79, p=0.003), moxifloxacin: (0.66, p=0.005), gatifloxacin: (0.37)] prior to
surgery. There were no differences in abs MRx diff or IOP between the three
fluoroquinolone groups. The NSAID groups_ clinical outcomes were not
statistically analyzed due to small sample sizes (ketorolac, n=76; bromfenac,
n=2; diclofenac, n=1). There were no cases of endophthalmitis or other
serious adverse events in any of the groups. 100% of subjects on gatifloxacin
had an increase in their personal cost over ofloxacin, while 22.5% had an
increase, 64.5% had no change, and 13.0% had a reduction in cost over
moxifloxacin. 100% of subjects who received ofloxacin and moxifloxacin
had cost savings over gatifloxacin. 100% of ketorolac users had cost savings
over bromfenac, while 20.5% had cost savings and 79.5% had no change
over diclofenac.
Conclusions: No significant differences were found in clinical outcomes for
subjects who received alternate agents versus preferred agents; however,
significant cost savings were noted for alternative agents.
112
89
VARIABLES ASSOCIATED WITH HOSPITAL READMISSION
OF PATIENTS WITH CONGESTIVE HEART FAILURE
Yusufzai M, Singh N, Sirohi R San Joaquin General Hospital, Stockton, CA.
Purpose of Study: Hospital readmission rates are high for patients with
Congestive Heart Failure (CHF). Hospitals are required to reduce 30-day
readmission rates. This study suggests that renal failure is an important factor
in readmission of CHF patients.
Methods Used: Two hundred and two patients were admitted to inpatient
service of a public hospital from June 2009 to June 2010 for CHF (males
n=138, females n=64). The medical records were reviewed in a retrospective
analysis to determine factors associated with readmission within 30 days.
There were age and sex matched controls (n=153) and study group readmitted for CHF (n=49). Chi-square and t tests were used for statistical
analysis.
Summary of Results: Out of 202 patients, 110 were readmitted with total
240 readmissions; of which 49 were readmitted for CHF with 82 readmissions. Out of the eighty-two, 41 readmissions were within 30 days.
Readmission is found related to many variables. Age, sex, smoking, alcohol
and drug abuse were non-significant risk factors. Renal failure was the most
significant factor for readmission within 30 days. Out of the 49 patients
who were readmitted for CHF, 69.4% (34/49) had renal failure compared to
44.4 %(68/153) of controls (pG0.002) (RR=2.22, CI=1.29Y3.82). Among
those readmitted within 30 days, 72.0% (18/25) had renal failure compared
to 47.5 %(84/177) of controls (pG0.018) (RR=2.52, CI=1.10Y5.77). Patients
with CHF readmissions had a lower left ventricular ejection fraction (LVEF)
and higher Serum creatinine levels as compared to controls (pG0.001).
Patients non-compliant with medications and those having non-commercial
state and county assistance insurance implying lower socio-economic status
had higher rates of readmissions (pG0.003). In the total group of 202 patients,
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
63% had diet restrictions, 23% CHF education and 42% cardiology clinic
follow-up appointments documentation in their medical records.
Conclusions: There is a significant increase in readmission rate in patients
with renal failure and the severity of renal failure may have prognostic significance. Follow-up appointment, diet, drug compliance and CHF education
should be given to all patients at hospital discharge. Special consideration
should be given to those with renal failure and limited access to health care.
90
EFFECT OF FAXING EMERGENCY DEPARTMENT
PRESCRIPTIONS ON PHARMACY WAIT TIME AND
SUBSEQUENT HEALTH CARE UTILIZATION
Fernando TJ, Baraff LJ David Geffen School of Medicine at UCLA, Los
Angeles, CA.
Purpose of Study: The primary objective of this study is to assess whether
faxing prescriptions directly to a pharmacy leads to decreased pharmacy wait
times and greater satisfaction with the process of obtaining medication.
Secondary objectives are whether faxed prescriptions are associated with
decreased return Emergency Department (ED) visits or primary care visits.
Methods Used: Patients discharged from the UCLA Emergency Department with prescription medications were approached for study consent.
Study participants were randomized into control or treatment groups. Participants in the control group were discharged with the standard prescription,
while participants in the treatment group had their prescriptions faxed to a
pharmacy. All study participants were contacted 7 days after ED discharge for
a telephone interview.
Summary of Results: Data analysis was performed on 60 patients
(23 males, 37 females) who completed the telephone interview. Study participants had a median age of 36 years (range=6Y86). 31 patients were prescribed 1 medication, 24 were prescribed 2 medications, 4 were prescribed
3 medications, and 1 was prescribed 4 medications. Five patients did not
pick up their medication from the pharmacy. Technical problems occurred
with 9 faxed prescriptions. Data collection is ongoing.
Conclusions: After blocking for technical difficulties that arose with faxed
prescriptions, we found a significant difference between the two groups in
pharmacy wait time (Wilcoxon p = 0.009) and whether medications were
ready for immediate pick up (Fischer_s exact G0.001). There was no significant difference in patient satisfaction with obtaining medication. Faxing
prescriptions did not significantly change the number of return ED or followup primary care visits.
Pharmacy Wait Time
p-value = 0.009
Percentage of Medications Ready For Pick Up
Fischer’s Exact = 0.000
91
LOWERING LIPOPROTEINS AMONG EMPLOYEES:
PROGRESS AT WORK
Raymond LW1,2, Pankowski J2, Shiflett H2 1Univ of North Carolina, Chapel
Hill, Charlotte, NC and 2Carolinas HealthCare System, Charlotte, NC.
Purpose of Study: Blood lipoproteins (BL) are an important reversible
determinant of cardiovascular diseases which can impact worker health, job
performance and health care costs. However, few reports have addressed the
impact of worksite health promotion (WHP) on BL levels. We evaluated the
effect of WHP on BL in employees of three companies which expressed
interest in this outcome.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Methods Used: Workers at a lumber yard (N=250), an electric co-operative
(N=150) and an engineering design firm (N = 115) were invited to participate
in free, non-fasting BL screening, as part of WHP. Serum analytes by autoanalyzer included total cholesterol (TCh) and high-density lipoprotein
(HDL), and triglycerides (TG). Low-density lipoprotein (LDL) was derived
using the Friedewald equation. Counseling on exercise, weight loss, dietary
optimization and smoking cessation were included in the WHP program, and
a physician sent each participant an individual letter with BL results, along
with advice on what to do about the results.
Summary of Results: 168 men and 24 women (mean ages 45 and 46 years;
37% of invitees) provided baseline BL samples, and follow-up samples were
available in 82 men and 16 women. TG and LDL were unavailable in only 5
of 196 blood samples with lipemic plasma due to hypertriglyceridemia. The
ratio of TCh to HDL exceeded 5.5 in 33 participants at baseline, and in
24 after 19 + 8SD months of follow-up. Significant reductions also occurred
in mean values of total, LDL and non-HDL cholesterol and in the ratio of
TCh to HDL.
Conclusions: WHP activities were associated with significant near-term
reductions in BL in these workers. Whether such reductions will be sustained
and be accompanied by less cardiovascular disease remains to be determined.
Non-participants remain a challenge.
Baseline and Follow-up Lipoproteins in 98 Workers at 3
Companies
92
AN EVALUATION OF THE ORAL HEALTH STATUS OF
RURAL UGANDAN SCHOOL CHILDREN ENROLLED IN
FBRIGHTER SMILES AFRICA_
Meredith C1, Berg T1, Ashaba B2, Choi W1, Zhang M1, Musinguzi N2,
Nambatya J2, Nyairo S2, Cannon W1, Kasangaki A2, Macnab A1 1University
of British Columbia, Vancouver, BC, Canada and 2Makerere University,
Kampala, Uganda.
Purpose of Study: To document the oral health status of a cohort of rural
Ugandan primary school children enrolled in an on-going school-based
health-promotion program FBrighter Smiles_, with reference to the distribution of caries and incidence of significant oral pathology.
Methods Used: A Ugandan/Canadian student/faculty team conducted the
program_s annual examinations using a validated dental survey (DMFS
Scores), and health questionnaires; oral pathology and atypical variation were
photographed, and comparison made of the number of anterior versus posterior teeth affected by caries and the extent of the decay. Education was given
to address the dental problems found.
Summary of Results: 294 children aged 5Y13 years were examined; 218
(74.2%) had oral pathology present. Anterior caries was present in 19 (6.5%)
and posterior caries in 95 (32.3%). Other significant atypical findings and oral
pathologies identified included gum recession in 4.4%, gingivitis in 12.9%
and calculus in 36.4%. Malocclusion, Treacher-Collins Syndrome, tetracycline staining, peg lateral incisors, mulberry molars, herpetic stomatitis,
candida, ankyloglossia, transposition and gemination were also present.
Workshops were delivered to teach better oral hygiene techniques for posterior teeth, and suggest health practices to improve other pathologies
where possible.
Conclusions: Posterior caries was more prevalent than anterior disease and
the range of pathologies extensive. Delivery of the Brighter Smiles Program
in rural schools is lowering the incidence of periodontal disease, reducing
caries, and promoting knowledge and healthy practices in a country with
limited health and dental care. The program is also providing valued collaborative educational and research opportunities for the teams involved.
113
Western Regional Meeting Abstracts
Journal of Investigative Medicine
93
REPETITIVE STRESS SYMPTOMS IN RADIOLOGISTS IN
THE DIGITAL AGE
Wood JP University of New Mexico, Albuquerque, NM.
Purpose of Study: With a switch to digital workstations, time spent in the
same position has increased significantly. PACS has increased efficiency, but
possibly at a physical cost to radiologists. This led to the development of this
pilot study: Evaluate the prevalence of repetitive stress symptoms by surveying radiologists in Albuquerque.
Methods Used: We sent our questionnaires to 92 radiologists throughout
Albuquerque (2 private practices, radiology residents and Attending
Radiologists at Univ. of New Mexico). We used EpiInfo and SAS v9.2 for
entry & analysis, and Fisher_s Exact Test due to small N-size.
Summary of Results: Overall, 58/92 responded (63%). 76% reported that
they experience pain (pain, stiffness, soreness, or cramping in extremity,
back, or neck). When response was Byes[ to experiencing pain, we asked
them to specify the type and severity (Likert Scale 1Y5, 5 being most severe).
The Mean ranged from 2.2Y2.4 for: Back, Neck, Arm/Shoulder, Wrist/Hand,
or Leg. Less than 9% reported some sort of ergonomic training in the past
year. When divided into private, academic attending, and resident, there was a
significant difference among the groups spending greater than eight hours per
day at a PACS/workstation (59%, 21%, 53% respectively). When comparing
these 3, academic Attendings reported the highest pain frequency of at least
once per month (86% compared to 76% in private and 67% in residents).
There was no difference amongst the other groups in age and pain frequency.
Conclusions: As a pilot study, repetitive stress symptoms were common.
The total N-size made statistically significant differences difficult, although
some observations were evident. There were enough interesting qualitative
findings (equal distribution of pain sites, no difference in pain correlating
with age) to make this study worth repeating with a larger population. Future
studies could include: the effect of documented ergonomic training on
improvement in symptoms, error rate-pain symptom correlation, or error
rate-break freq correlation.
94
MISSION, SENSEMAKING, AND TOOLS: EXPLORING THE
CAPACITY OF COMMUNITY HEALTH CENTERS TO
IMPLEMENT EVIDENCE-BASED INTERVENTIONS
Terasaki D 1UW School of Medicine, Seattle, WA and 2University of
Washington, Seattle, WA.
Purpose of Study: Community health centers (CHCs) make up a large
segment of the health care safety net in the United States. Due to their patient
demographics and placement in underserved areas, CHCs are well positioned
to decrease health disparities through evidence-based interventions. A multidisciplinary research group has developed a conceptual framework (Practice
Change model) that is useful for exploring change in primary care settings.
A section of the model labeled BAdaptive Reserve,[ which includes 7
characteristics of healthy work relationships, held particular interest to our
research team. We aimed: 1) To assess the applicability of the Practice
Change model to CHCs, focusing on Adaptive Reserve, and 2) To gain
insight into the implementation of evidence-based programs at CHCs.
Methods Used: Key informant interviews of administrative and clinical
leaders were conducted at 4 CHCs and 1 national association. Interviews
were recorded and transcribed verbatim. Three research personnel independently coded each transcript and then convened to discuss emerging
themes. The final set of codes was subsequently entered in Atlas.ti for
further analysis.
Summary of Results: Findings from 14 interviews elucidated 3 domains in
the Practice Change model - inside/outside motivators, dynamic local ecology, and sensemaking - that merited special emphasis with regards to CHCs.
In contrast to mainstream practices, for CHCs: 1) their mission serves as a
major inside/outside motivator; 2) their dynamic local ecology is greatly
influenced by the communities they serve; and 3) tools that facilitate sensemaking and key implementation steps enhance their capacity for change.
Informants underscored BTrust[ as the most significant work relationship and
described their work place as a Bfamily-like community.[
Conclusions: Our findings support the importance to design interventions in
partnership with CHCs that: 1) align with CHC_s mission and investment in
their community; and 2) utilize effective tools to aid in various stages of
114
&
Volume 59, Number 1, January 2011
implementation, including sensemaking. This may achieve greater buy-in
from CHC personnel and, ultimately, facilitate the implementation of
evidence-based interventions that reduce health disparities where the
need is greatest.
95
TRANSPARENT REPORTING OF GAMBLING
EFFECTIVENESS STUDIES
Parhami I, Campos MD, Fink A, Fong T University of California - Los
Angeles, Los Angeles, CA.
Purpose of Study: Gambling disorders are a significant health concern
because they are associated with cardiovascular disease, major depression,
substance abuse, financial troubles, and crime. This study systematically
reviews the transparency of reports on interventions to ameliorate the adverse
consequences of problem and pathological gambling. Transparent reporting
is essential to the assessment of study quality.
Methods Used: Using specific inclusion and exclusion criteria, we identified 25 articles from Pub Med, PsycINFO, and the Web of Science published
between January 2000 and August 2010. Two researchers independently
reviewed each article_s reporting using 58 items adapted from the American
Public Health Association_s and CDC_s TREND (Transparent Reporting of
Evaluations of Nonrandomized Designs) Statement. Twenty-two items represent particularly important criteria, and all should be reported. Differences
in interpretation between reviewers were adjudicated by a third reviewer.
Summary of Results: Only 8 articles met at least 70% of the 58 general
reporting criteria, (median= 64%; range: 41Y84 %). None of the studies met
all 22 of the important reporting criteria. Examples of particularly important
criteria met by at least half the studies include descriptions of eligibility rules
for participation, methods for handling missing data, and methods for
establishing baseline study group equivalence. Fewer than half the studies
described the barriers encountered in implementing the experimental interventions, methods for increasing compliance, and methods for addressing
differences in baseline characteristics of participants who were lost to followup compared to those who were retained. No differences in transparency
were found between articles published before and after the initial dissemination of the TREND criteria (2004).
Conclusions: Published studies reporting on the effectiveness of interventions for gambling related disorders tend to vary in their transparency
with many important reporting criteria unmet. Transparent reporting is a
prerequisite for determining if evidence is available to support implementing specific programs or treatments to address gambling disorders
and the untoward medical and social consequences with which they are
associated.
Hematology and Oncology I
Concurrent Session
8:30 AM
Friday, January 28, 2011
96
SURVIVAL OUTCOMES OF ADULTS WITH ACUTE MYELOID
LEUKEMIA-A STUDY OF NATIVE AMERICAN, HISPANIC,
AND NON-HISPANIC WHITES TREATED IN NEW MEXICO
Armijo B , Libby E , Quintana D , Rivera N , Wiggins C , Lee S , Abdul-Jaleel M
University of New Mexico, Albuquerque, NM.
Purpose of Study: The goal of this study is to identify potential differences
in outcome between ethnic groups from urban or rural areas in patients with
newly diagnosed acute myelogenous leukemia (AML) treated at this center
from 1986-2006.
Methods Used: A retrospective chart review was performed on patients
identified through a New Mexico tumor registry search of all adult patients
(Q 18 years of age) with a new diagnosis of AML treated at this center
from 1986 through 2006. 144 charts were available for review. Electronic,
microfilm and paper-based medical records were reviewed. Data was collected for race/ethnicity, sex, zip code, age at diagnosis, pathologic diagnosis,
date of diagnosis, cytogenetics, treatments received, total number of chemotherapy cycles, achievement of complete remission, and overall survival
from time of diagnosis and from achievement of remission after induction
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
chemotherapy. Cancer cases who resided in New Mexico counties with large
population centers were designated as urban residents and the remaining
cases were designated as rural residents. Statistical analysis was performed
to examine the effect of the variables mentioned above on overall survival.
Summary of Results: The analysis included 144 patients (mean age 49.98,
range 18-85). For those patients who received chemotherapy the standard
treatment was 7+3 followed by high dose cytarabine (1Y4 cycles) or similar
regimen. There were no statistical differences between survival rates when
comparing the three ethnic groups, Caucasians 78 (52%), Hispanics 43
(28%), and Native Americans 23 (15%) (P=0.2496). In agreement with
previous reports, statistically significant differences were seen for treatment
versus palliative care, cytogenetics and age. The study included 61 urban
patients with a mean survival of 426 days and 83 rural patients with a mean
survival of 278 days. Analysis revealed a statistically significance in survival
for urban versus rural patients who achieved complete remission after
induction chemotherapy (P=0.0443).
Conclusions: Survival was improved for urban patients who achieved a
complete remission after induction chemotherapy versus rural patients
(P= 0.0443). Further study should be undertaken to identify the causes
of this disparity.
97
VALUE OF CONTROL GROUPS IN PHASE 2 CLINICAL
TRIALS
Moseley J1,2, Pagel J2,1, Othus M2 1University of Washington, Seattle, WA
and 2Fred Hutchinson Cancer Research Center, Seattle, WA.
Purpose of Study: Each year, numerous abstracts describing new therapies
for acute myeloid leukemia (AML) are presented at the American Society
of Hematology (ASH) meeting. 63/91 abstracts submitted from 1993Y2006
suggested therapeutically promising results; these 63 abstracts covered 37
drugs. However, 34/37 (92%) drugs initially viewed positively at ASH were
found to have negative results with further study or have disappeared from the
clinical trials arena. Walter et al note that absence of a control group contributes to this preponderance of false-positive, misleading results.
We encountered a specific example in a single-arm trial of a 4-drug
combination (FLAM) investigated at Johns Hopkins and UW/FHCRC in 72
patients with Bpoor prognosis[, newly-diagnosed AML. The initial conclusion was that Bthe salutary results of FLAM in poor-risk pts will be extended
to adults with non-poor risk AML and compared with traditional cytotoxic
chemotherapy.[ Our aim was to determine if the same held true when a
control group was included in the analysis.
Methods Used: We identified a control group of 229 newly-diagnosed
patients given standard therapy at either UW/FHCRC or in Southwestern
Oncology Group trials. We next collected information on relevant prognostic
factors in this group: age, performance status, cytogenetic findings, and
whether disease onset was immediate or delayed. Finally, we combined the
FLAM and control patients and used multivariate analysis to examine
whether, after accounting for possible differences between FLAM and control in the distribution of prognostically-relevant covariates, the rate of
complete remission (CR) was higher with FLAM.
Summary of Results: The multivariate analysis suggests that the probability that FLAM is 910% superior to standard therapy is G 1%. Additionally,
FLAM does not appear more effective in any particular prognostic subgroup.
Conclusions: While not a substitute for a randomized trial comparing standard therapy and FLAM, our analysis indicates it is highly unlikely that FLAM
is meaningfully superior to standard therapy. Our results motivated cancellation
of a planned randomized trial. This will enable other drugs to be studied rather
than devoting patients and other resources to further study of FLAM.
98
SURVIVIN AS A POTENTIAL SENSOR OF OXIDATIVE
STRESS IN HUMAN BREAST CANCER
Pervin S1,2, Tran A1, Tran L1, Urman R1, Chaudhuri G1, Singh R1,2 1Charles
Drew University of Medicine and Science, Los Angeles, CA and 2UCLA,
Los Angeles, CA.
Purpose of Study: Breast cancer remains a poorly understood heterogeneous disease. Survivin, a unique member of the inhibitor of apoptosis (IAP)
gene family, is over expressed in aggressive tumors and its down regulation
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
sensitizes cancer cells to various therapeutic agents. The purpose of this study
is to examine the role of survivin as a potential sensor of oxidative stress
in human breast cancer.
Methods Used: Oxidative stress was induced in a panel of breast cancer cell
lines by treating with DETA-NONOate (1-3mM), a nitric oxide (NO) donor
and expression of MAP Kinase Phasphatase-1 (MKP-1), an early stressresponse gene, was examined. Survivin levels were either up regulated by
over-expression of full-length human survivin cDNA or inhibited using small
inhibitory RNA (siRNA) to examine the sensitivity of cells to oxidative
stress. Relationship between survivin and MKP-1 was also examined in fresh
and paraffin sections of human breast tumors by quantitative western blot,
real-time PCR and immunohistochemistry.
Summary of Results: NO-induced oxidative stress down regulated cytosolic survivin and induced MKP-1 expression in estrogen receptor negative
(ER-) breast cancer cell lines. This down regulation of cytosolic survivin
protein was an early event and preceded the induction of MKP-1. High level
of oxidative stress was required to induce MKP-1 in survivin over-expressing
cells. In addition, reduction of basal survivin levels rendered these cells
sensitive to low oxidative stress. However, ER+ cell line had low levels of
only nuclear survivin but high MKP-1 levels. We further demonstrate that
ER+ fresh human breast tumors also had low survivin, but high levels
of MKP-1 expression. This inverse relationship between survivin and
MKP-1 was also detected in fresh and paraffin-sections of ER- human
breast tumors.
Conclusions: Reduction of survivin levels was essential for sensitizing the
breast cancer cells to stress by upregulating MKP-1. High levels of survivin
render ER- aggressive breast tumors insensitive to oxidative stress. We
therefore, conclude that survivin levels determine the sensitivity of human
breast cancer cells to NO-induced oxidative stress.
99
RASSF1C OVER-EXPRESSION PROTECTS LUNG CANCER
CELLS FROM BETULINIC ACID EFFECTS ON CELL
PROLIFERATION
Jo J, Franke E, Amaar Y, Reeves M Loma Linda University, Loma Linda, CA.
Purpose of Study: Ras association domain family 1 (RASSF1) gene is a
Ras effector that plays an important role in carcinogenesis. The gene encodes
two major proteins derived from alternate mRNA splicing. RASSF1A is a
tumor suppressor. The function of RASSF1C remains to be fully elucidated.
Our lab has evidence suggesting RASSF1C may promote lung cancer cell
growth. We showed that RASSF1C over-expression increases proliferation of
non-small cell lung cancer (NSCLC) cells, NCI-H1299. We discovered that
RASSF1C stimulates expression of interesting genes including the stem cell
self-renewal gene PIWIL1 (also known as HIWI) in NSCLC cells.
Betulinic Acid (BA) has been shown to inhibit cell proliferation and
migration, induce apoptosis in different human cancer types including lung
cancer, and to down regulate PIWIL1expression in gastric adenocarcinoma.
In light of these findings and our findings on RASSF1C, we wish to determine if the down regulation of PIWIL1 gene expression and anti-cancer
activities exerted by BA can be reduced by RASSF1C over-expression.
Methods Used: Cell proliferation: NCI-H1299 was transduced with MLV
back-bone vector (NCI-BB) as a control or RASSF1C (NCI-1C) gene. Cells
were cultured, counted, and plated onto 96 well plates. The cells were
acclimated to serum-free media and exposed to 20Y50Kg/ml BA for 24 hr,
and then analyzed by Alamar Blue assay. A plate reader analyzed fluorescence levels to indirectly measure cell proliferation or suppression.
RT-PCR: Cells were plated in 6-well plates and treated with 20-50Kg/ml
BA for 24 hr and then collected. RNA was isolated and used for RT-PCR
analysis using RASSF1C and PIWIL1 gene specific primers.
Western Blot analysis: Cells were plated in 10 cm plates and treated with
20Y50Kg/ml BA for 24 hr. Cells were collected and used to prepare cell
lysates for Western blot analysis using PIWIL1-antibody.
Summary of Results: Over-expression of RASSF1C seems to provide
NCI-H1299 lung cancer cells with protection against the actions of BA.
RT-PCR and Western blot analysis show that BA does not down-regulate
PIWIL1 gene expression in NCI-H1299 cells.
Conclusions: RASSF1C over-expression reduces the anti-cancer activities
exerted by BA on lung cancer cell proliferation. BA does not affect PIWIL1
gene expression in lung cancer cells unlike what has been reported in gastric
adenocarcinoma cells.
115
Western Regional Meeting Abstracts
Journal of Investigative Medicine
100
THE PENTOSTATIN PLUS CYCLOPHOSPHAMIDE (PC)
NON-MYELOABLATIVE REGIMEN INDUCES DURABLE
HOST T CELL FUNCTIONAL DEFICITS AND PREVENTS
MARROW ALLOGRAFT REJECTION
Taylor J1,2, Mariotti J2, Fowler DH2 1University of New Mexico School of
Medicine, Albuquerque, NM and 2National Cancer Institute, Bethesda, MD.
Purpose of Study: Although pentostatin [P] and fludarabine [F] have each
been incorporated into non-myeloablative regimens, it is unknown whether
these purine analogs differentially modulate host immunity prior to
transplantation.
Methods Used: To address this, we utilized murine models to directly
compare pentostatin and fludarabine for their ability to: (1) operate synergistically with cyclophosphamide [C] to induce host T cell depletion; (2)
induce host T cell suppression, as defined by modulation of cytokine
secretion in vitro and abrogation of host-versus-graft reactivity (HVGR) in
vivo; (3) constrain host T cell recovery post-chemotherapy; and (4) prevent
the rejection of T-cell depleted (TCD), fully MHC mismatched bone marrow
allografts.
Summary of Results: Relative to single-agent regimens, combination PC
and FC regimens, which were given over 3-days or 14-days, worked synergistically to deplete host CD4+ and CD8+ T cells; PC and FC regimens were
developed that yielded similar levels of host T cell and myeloid cell depletion.
In the setting of these generally comparable states of host T and myeloid cell
depletion, the PC regimen was found to be more immune suppressive, as
evidenced by reduced host T cell capacity to: (1) secrete IL-2 and IFN-F in
vitro; (2) mediate HVGR in vivo; and (3) recover numerically and functionally during a two-week observation period post-chemotherapy. Finally,
using B6 hosts treated with the 14-day chemotherapy regimens, the PC
regimen more consistently prevented the rejection of BALB/c TCD-allografts
than the FC regimen (rate of alloengraftment, 14/15 [93%] of PC-treated
recipients vs. 8/14 [57%] of FC-treated recipients; pG0.05).
Conclusions: Pentostatin and fludarabine were thus similar in their ability to
operate synergistically with cyclophosphamide to induce marked host T cell
depletion. However, the pentostatin-based regimen more profoundly suppressed host T cell cytokine potential, alloantigen-driven clonal expansion,
numerical and functional recovery, and ability to reject fully MHC-disparate
marrow allografts.
&
Volume 59, Number 1, January 2011
Mutation of these binding sites on the survivin promoter showed abrogation
of the observed repression of survivin promoter activity.
Conclusions: These data suggest that YY1 is a novel repressor of survivin
transcription. A better understanding of how YY1_s regulation of survivin
expression in vivo affects cell cycle and cell death may provide new directions in therapeutics designed to prevent survivin overexpression in cancer.
102
SLUG MRNA IS HIGHLY EXPRESSED IN SQUAMOUS LUNG
CARCINOMA AND ASSOCIATED WITH EXPRESSIONS OF
EGFR AND ZEB1 IN PATIENTS WITH NON-SMALL CELL
LUNG CANCER (NSCLC)
Zhao JM, Yoshida K, Mascaux C, Wynes M, Dziadziuszko R, Witta S,
Hirsch F University of Colorado at Denver, Denver, CO.
Purpose of Study: Understanding biological markers for disease progression and metastasis in lung cancer is a key step for developing new treatment.
The Epithelial-to-Mesenchymal Transition (EMT), a mechanism involved in
the processes of invasion and metastases and sensitivity to EGFR Tyrosine
Kinase Inhibitors, is characterized by the upregulation of the transcription
repressors slug (a member of the snail family) and ZEB1 (Zinc finger E-box
Binding homeobox 1) and the resulting downregulation of E-cadherin.
Methods Used: In the present study, we performed quantitative RT-PCR
analysis on resected tumors from 121 patients with NSCLC to measure the
mRNA expressions of slug, ZEB1 and EGFR using beta-actin as the normalization signal.
Summary of Results: Slug mRNA was found to be significantly higher
expressed in squamous lung carcinoma as compared with adenocarcinoma
with a mean level of expression of 1.41 vs. 0.61 (p = 0.0009), respectively.
In addition, slug was positively correlated with EGFR (Spearman r = 0.33,
p = 0.0011) as well as with ZEB1 (Spearman r = 0.24, p = 0.028).
Conclusions: Our results demonstrate that slug mRNA is more elevated in
squamous lung carcinoma, indicating that slug could be a potential target for
the treatment of this type of lung cancer. The positive correlation between
slug and EGFR based on patient tumors are consistent with the previous cell
line data that showed decreased expressions of slug and ZEB1 when EGFR
pathways were inhibited. Our results offer new insights into the pathways of
EMT activation. and suggest a synergistic relation between EGFR activation,
its downstream Myc product in the MEK/ERK pathway, and slug induction.
101
YIN YANG 1 REGULATES THE TRANSCRIPTIONAL
REPRESSION OF SURVIVIN
103
DETECTION OF INFLAMMATORY BIOMARKERS IN THE
BLOOD OF PATIENTS WITH BREAST CANCER METASTASIS
Galloway N1, Li F2, Sui G3, Wall N1 1Loma Linda University School of
Medicine, Loma Linda, CA; 2Roswell Park Cancer Institute, Buffalo, NY and
3
Wake Forest University School of Medicine, Winston-Salem, NC.
Purpose of Study: Survivin is a member of the Inhibitor of Apoptosis
(IAP) family of proteins, and is highly expressed in all cancers but absent in
normal tissue. Expression level correlates with chemo- and radioresistance,
as well as poor prognosis in cancer patients. The mechanisms for upregulation of Survivin in cells undergoing stress associated with tumor development
and the tumor microenvironment are not well understood. The putative stress
response transcription factor Yin Yang 1 (YY1) was hypothesized to contribute to the upregulation of survivin in tumor cells.
Methods Used: In order to study regulation with luciferase reporter assays,
U2OS cells were transfected with nested deletions of the survivin promoter,
ranging from short (+230 bp) to long (+6280 bp). YY1 involvement in
survivin promoter repression was confirmed using siRNA directed against
YY1. A U2OS cell line containing a stable YY1 Tet-off system was used to
determine whether a temporal increase in YY1 expression affects Survivin
protein levels. To further evaluate the role of YY1 regulation of survivin
expression, reporter constructs containing mutated putative YY1 binding
sites in the proximal survivin promoter were used in reporter assays.
Summary of Results: When YY1 was overexpressed, luciferase expression
was repressed 5Y10 fold. Further studies showed that knockdown of YY1
releases the survivin promoter from the observed repression and leads to a
3Y5 fold increase in promoter activity above basal levels. Furthermore, a
low to moderate decrease in Survivin protein was observed in Tet removal
experiments. Site-directed mutagenesis confirmed the involvement of proximal survivin promoter sites in YY1 overexpression-induced reporter repression.
Mahuron K1, Foubert P2, Natarajan L3, Pierce JP4, Varner J2,1 1University of
California, San Diego, La Jolla, CA; 2University of California, San Diego,
La Jolla, CA; 3University of California, San Diego, La Jolla, CA and
4
University of California, San Diego, La Jolla, CA.
Purpose of Study: Breast cancer is the most common type of tumor and the
second leading cause of cancer-related death for women in North America.
The main cause of these deaths is not the primary tumor, but rather is from
distant metastases. While standard treatment modalities have improved the
overall outlook for women with breast cancer, there is still a need for better
therapeutic and diagnostic approaches for patients that face metastasis.
Clinical and experimental data have shown that pro-inflammatory cytokines
and systemic inflammation can facilitate tumor metastasis, suggesting a
correlation between inflammation and cancer metastasis. Therefore, the aim
of this study is to detect new inflammatory biomarkers in the blood, which
would potentially lead to earlier diagnoses and more successful outcomes for
breast cancer patients with metastasis.
Methods Used: We evaluated the relationship between blood inflammatory
biomarkers and breast cancer recurrence using blood samples from the
Women Healthy Eating and Living (WHEL) study at UCSD. We obtained 20
case-control pairs. The cases are women who had distant metastasis. Controls
are matched to cases based on date of diagnosis and stage of the primary
breast cancer tumor and age at diagnosis of the primary cancer. Blood
samples were acquired from both case and control women 1Y2 years after
their primary tumors were resected and before any signs of metastasis were
present in the case women. RNA was extracted from buffy coats in order
to perform quantitative RT-PCR to evaluate expression of selected inflammatory markers.
116
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: VEGF-C gene expression was higher in buffy coats
from patients with breast cancer metastases versus controls (p=0.01) after
primary tumor resection and before evidence of metastasis.
Conclusions: Our results indicate that VEGF-C might be used as a
biomarker for the discrimination of breast cancer patients who will have
recurrent metastasis and should receive more aggressive therapies.
104
INFLAMMATORY MEDIATORS REGULATE IMP-3
EXPRESSION IN HEAD AND NECK SQUAMOUS CELL
CARCINOMA (HNSCC)
Anastasiou C1, Luo J2, St. John M2 1David Geffen School of Medicine at
UCLA, Los Angeles, CA and 2UCLA Health Center, Los Angeles, CA.
Purpose of Study: Insulin-like growth factor-II mRNA binding protein 3
(IMP-3; also known as homology domain-containing protein over-expressed
in cancer and L523S), is an oncofetal protein believed to regulate translation of the potent growth factor and apoptosis inhibitor, insulin-like growth
factor-II. IMP-3 is over-expressed in many HNSCCs and correlates with
higher histologic grade, lymph node metastases (LNM), and advanced stages
of disease.
HNSCCs are highly inflammatory and notably aggressive cancers. Prolonged exposure of human neoplasms to inflammatory mediators has been
shown to promote epithelial-mesenchymal transition (EMT), tumor growth,
invasion, angiogenesis, genetic instability, and metastasis. Among the myriad
of inflammatory mediators elevated in the HNSCC microenvironment and
associated with EMT and tumor growth is interleukin-1 (IL-1). We examined
the effects of IL-1A on IMP-3 transcription and translation.
Methods Used: Western blot and real-time reverse-transcriptase polymerase
chain reaction (RT-PCR) were utilized to determine how IL-1A affects IMP-3
expression in two well-characterized HNSCC lines Tu212 and Tu686, and in
NFkB knock-down cells.
Summary of Results: IL-1A treated HNSCC cells showed significantly
increased IMP-3 protein expression. This effect was diminished when NFJB
knockdown cells were treated with IL-1A.
Conclusions: Our study demonstrates that inflammatory mediators in the
tumor microenvironment increased IMP-3 expression in HNSCC. This is the
first evidence that IL-1A may play a role in IMP-3 regulation, and that NFkB
may be an intermediate in this regulatory pathway. Unraveling pathways in
which IMP-3 operates may help identify novel targets for use in cancer
prevention and therapy.
105
IS RACE A DETERMINANT OF SURVIVAL IN PATIENTS
WITH CASTRATION RESISTANT PROSTATE CANCER
TREATED WITH DOCETAXEL?
Ravi-Kumar S, Verschraegen C, Lee S University of New Mexico,
Albuquerque, NM.
Purpose of Study: The aim of this study was to identify ethnic differences
between Hispanics(H), Whites (NHW), and Native Americans (NA) with
Castration Resistant Prostate Cancer (CRPC) in response to Docetaxel.
Methods Used: After Institutional Review Board approval, medical records
of 59 CRPC patients treated with Docetaxel between 1999Y2010 at the
University of New Mexico Cancer Center and Hematology Oncology
Associates were reviewed. Collected data included race, age, prior treatments,
site of disease, and Gleason score. Docetaxel outcome variables were concurrent chemotherapy; baseline, nadir, and 12 week PSA; number and toxicity of Docetaxel courses; and addition of other treatments to Docetaxel.
Progression of disease was defined as the duration of treatment with
Docetaxel before the identification of a new lesion (by RECIST criteria)
or PSA progression. Primary end point was overall survival (OS). Secondary endpoints was progression free survival(PFS).
Summary of Results: Median age of our cohort of patients was 71 years.
There were 34 NHW, 16 H, 5 NA, 1 each of Asian, Black and Unknown. The
median PFS was 24.6 weeks. The median overall survival was 44 weeks. The
Gleason score at diagnosis did not have a statistically significant effect on OS
or PFS of these patients. The OS was highest among NHW (75 weeks),
followed by NA and worst for H (42.4 weeks). However, these results were
not statistically significant (p value=0.2778). PFS was greatest for NHW,
followed by NA and then H (not statistically significant with a p value of
0.2778). significant). Statistically positive variables for survival included
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
PSA values at all time points, number of docetaxel courses and level of
neutropenia following the first course of Docetaxel. The addition of
thalidomide to Docetaxel had a favorable effect both on OS and PFS
(pG0.0001 and pG0.0384, respectively).
Conclusions: In conclusions, ethnicities other than Black do not influence
outcome of patients with CRPC, but known variables, such as PSA values and
dose of chemotherapy did. Most interestingly, the addition of thalidomide
significantly improved outcome in all New Mexican ethnicities.
106
DO WE KNOW WHAT CHILDREN RECEIVING LEUKEMIA/
LYMPHOMA THERAPY HAVE VARICELLA ZOSTER IGG
ANTIBODY PROTECTION?
Wolff L Doenbecher Children’s Hospital/Oregon Health & Science
University, Portland, OR.
Purpose of Study: Varicella Zoster (VZ) infection can still be a life
threatening illness in patients (pts) receiving anticancer treatment (Tx).
Difficulty in determining the VZ antibody (Aby) status of pts led to the
question: Do we know what pts have VZ Aby protection (Pro) at diagnosis
(Dx) and during Tx?
Methods Used: A review of the records of all pts under 21 years who were
treated for leukemia or lymphoma (L/L) at Doernbecher Children’s Hospital
between 1/1/2003 and 12/31/2005 was done. Pt activity was followed for at
least 4 years. Data reviewed included Dx, age, Tx, VZ Aby Pro at Dx, VZ
Aby status (at least 4 months after onset of Tx) and clinical course.
Summary of Results: 118 were studied. At Dx, 73 pts were tested for VZ
Aby; 45 were not. 49 pts had Aby Pro. 25 pts were tested at least 4 months
after Tx was started: 7 pts Aby Pro at Dx were Aby Pro, 6 pts not tested at Dx
were Aby Pro, 12 pts were Aby negative. 33 pts were never tested. Review of
immunizations recorded was not helpful. 2 pts developed Varicella: one pt
was never tested and the other Aby negative. 2 patients developed Zoster.
Conclusions: The VZ Aby status at Dx was known in 73 L/L pts (62%).
The VZ Aby status, 4 or more months after Dx was known in 25 L/L
pts (21%).
The VZ Aby status of 33 (L/L) (28%) pts was never known.
During L/L Tx, there is a significant population of pts, whose VZ Aby
status is not known.
Metabolism I
Concurrent Session
8:30 AM
Friday, January 28, 2011
107
PREVALENCE OF METABOLIC SYNDROME (METS) IN A
MALE VETERAN CIGARETTE SMOKING POPULATION
DeMoss D1, Rolle PD1, Ferry L 2,1, Tonstad S 1, Fireck A 2,1 1Loma Linda
University, Loma Linda, CA and 2Loma Linda VA Medical Center, Loma
Linda, CA.
Purpose of Study: We have previously found that MetS is present in nearly
one half of our local Veteran population at the Loma Linda Veterans
Administration Medical Center. Smoking has been reported to be casually
associated with MetS and may therefore contribute to this high prevalence
in Veterans. We determined the proportion of smokers that met the criteria
for MetS. In addition to waist circumference, BMI was used as a measure of
central obesity in an attempt to examine the utility of BMI in comparison to
the gold standard of waist circumference as a criterion for MetS.
Methods Used: We accessed computerized patient record systems for
500 consecutive male patients enrolled in the BBreak the Chains[ smoking
cessation class (August 2008Y June 2009). We used the harmonized definition
of MetS as published in Circulation (2009) for 239 patients (any 3 criteria
of triglycerides 9150 mg/dL, HDL cholesterol G40 mg/dL, systolic BP 9 130
and/or diastolic BP 985 mmHg, fasting plasma glucose 9100 mg/dL and waist
circumference 940 inches*). For all 500 patients we used BMI 930 kg/m2 as
an alternative to waist circumference to evaluate the diagnostic utility of using
BMI**.
Summary of Results: The prevalence of MetS in the subject populations was 57.2% using the BMI criterion and 63.2% using the waist circumference criterion. Of the 239 subjects with both measures, the sensitivity
117
Western Regional Meeting Abstracts
Journal of Investigative Medicine
and specificity to detect an increased central obesity using waist circumference as the gold standard was 80.8% and 84.9%, respectively.
Conclusions: Smokers are at high risk population for MetS and may gain
weight after stopping smoking. While, waist circumference as a measure of
central obesity identified a higher prevalence of MetS than BMI, sensitivity
and specificity of BMI were high in this population of smokers and should
be considered as an alternative measurement. Smokers need to have more
aftercare for chronic disease risk management after achieving long term
abstinence.
* Any of the above components of the metabolic syndrome could be
satisfied alternatively by drug treatment or recorded clinical diagnosis.
**A BMI of 30 was chosen based on the WHO 1998 classification of
MetS criteria.
108
TESTOSTERONE DEPRIVATION ACUTELY DIMINISHES
INSULIN SENSITIVITY IN YOUNG, HEALTHY MEN
Rubinow K, Snyder C, Amory J, Bremner W, Page ST University of
Washington, Seattle, WA.
Purpose of Study: In men with prostate cancer, androgen deprivation
therapy (ADT) alters glucose metabolism. Whether sex steroids modulate
insulin sensitivity in healthy men remains unclear, as does whether such
effects result from changes in testosterone or its active metabolite estradiol.
We evaluated the effects of acute sex steroid withdrawal on insulin sensitivity
in healthy men and determined the relative contributions of testosterone (T)
and estradiol (E) to these effects.
Methods Used: We recruited 24 healthy men, 18Y55 years old, with normal
baseline T levels. All subjects received the GnRH antagonist acyline
(300 mcg/kg/2 weeks 2). In addition, subjects were administered either
placebo transdermal gel and pills (Group 1), transdermal T gel 10g/day plus
placebo pills (Group 2), or transdermal T gel plus the oral aromatase inhibitor
anastrozole 1 mg/day (Group 3) daily for 28 days. Body weight, fasting
serum hormones, adiponectin, glucose and insulin levels were measured
bi-weekly.
Summary of Results: Group 1 subjects were medically castrate with
treatment (T=0.8T0.8 nmol/L, E=32T11 pmol/L) whereas normal, baseline
sex steroid levels remained unchanged in Group 2 subjects. Group 3 subjects
maintained normal T levels while their serum E levels were equivalent to
those in Group 1 (E=37T14 pmol/L). Group 1 subjects exhibited no changes
in body weight or fasting plasma glucose, but their fasting insulin concentrations increased with treatment (P=0.02 vs. baseline) as did HOMA-IR
(P=0.04 vs. baseline). Interestingly, adiponectin levels substantially increased
in Group 1 (P=0.004 vs. baseline). None of these metabolic changes were
observed among subjects in Groups 2 and 3.
Conclusions: Our data demonstrate that acute T withdrawal reduces insulin
sensitivity independent of changes in body weight. Moreover, in the setting of
androgen deprivation, increases in adiponectin do not exhibit the expected
association with enhanced insulin sensitivity, thereby suggesting that adiponectin may be regulated directly by T. Changes in insulin sensitivity were
not evident among subjects with a selective reduction in estradiol, indicating
that diminished insulin sensitivity was attributable to androgen deprivation.
Whether androgen replacement in hypogonadal men can improve insulin
sensitivity merits further investigation.
109
PROTECTIVE EFFECT OR TESTOSTERONE AGAINST
HIGH FAT DIET INDUCED NON-ALCOHOLIC FATTY
LIVER DISEASE IN T-DEFICIENCY RATS MAY NOT BE
INSULIN-DEPENDENT
Jia Y1, Nikolaenko L1, Wang C1, Chong C1, Diaz-Arjonilla M1, Laurel S1,
French S2, Yee JK1, Lee PW1, Lue Y1, Swerdloff RR1 1LA BioMed,
Torrance, CA and 2LA BioMed, Torrance, CA.
Purpose of Study: NAFLD is associated with obesity, diabetes, insulin
resistance (IR) and Metabolic Syndrome (Met S). Testosterone (T) deficiency
is a risk factor for developing diabetes and Met S, but the role of testosterone
deficiency in hepatic steatosis has not been well studied. Our laboratory has
shown that high fat diet (HFD)-induced NAFLD was attenuated by T replacement in castrated rat. In this study we examined whether T amelioration
of NAFDL is related to changes in IR. We studied the 4 key biomarkers of
insulin action including adipose triglyceride lipase (ATGL) and hormone
118
&
Volume 59, Number 1, January 2011
sensitive lipase (HSL) and sterol regulatory element-binding protein-1
(SREBP-1) and fatty acid synthase (FAS).
Methods Used: Male rats were randomly placed into four groups: castrated
rats on high-fat diet (HFD), castrated rats with T replacement on HFD, intact
rats on HFD, and intact rats on regular chow diet (RCD). The HFD provided
71% energy from fat; RCD provided 16% of energy from fat. The rats were
fed ad libitum for 15 weeks then animals were killed and liver tissue collected and kept at j80C. Western blot was used to determine the changes in
expression of ATGL, HSL, SREBP-1 and FAS.
Summary of Results: As previously reported (Nikolaenko et al. ENDO
2010), serum T levels were not detectable in castrated rats, and T replacement
led to higher serum T levels than in intact rats. No statistical difference was
detected in serum glucose or insulin level between groups. The four insulinregulated proteins, ATGL, HSL, SREBP-1, and FAS were not changed with
castration after T treatment in rats fed a high fat diet.
Conclusions: As we reported previously, T treatment in castrated rats fed
a HFD reduces hepatic steatosis and fibrosis compared with castrated animals. Insulin level and insulin-regulated proteins were not changed after
T replacement. Our data suggest that androgen deficiency contributes to
the severity of hepatic steatosis and that T plays a protective role in liver fat
accumulation and NAFLD development. Insulin resistance may not play an
important role in this rat model of HFD-induced NAFLD and the protective
effect of T in reversing NAFLD.
110
RAPAMYCIN ADMINISTRATION IMPROVES METABOLIC
ABNORMALITIES AND DECREASES ADIPOSE TISSUE
INFLAMMATION IN DIET-INDUCED OBESITY
Subramanian S, Goodspeed L, Wang S, O’Brien KD, Chait A, Ladiges W
University of Washington, Seattle, WA.
Purpose of Study: Obesity is often associated with insulin resistance,
which may promote and increase the risk of cardiovascular disease. However,
the exact molecular mechanisms by which obesity leads to insulin resistance
are poorly understood. A hallmark of visceral obesity is the accumulation
of adipose tissue macrophages which can contribute to insulin resistance.
Aberrant, chronic activation of the intracellular nutrient sensing protein,
mammalian target of rapamycin (mTOR), a downstream effector of the
insulin signaling pathway, has been implicated in the pathogenesis of insulin
resistance. We hypothesized that oral administration of rapamycin, a potent
and specific inhibitor of mTOR, could reduce insulin resistance and improve
adipose tissue macrophage accumulation.
Methods Used: C57BL/6J mice were placed on a high fat, high carbohydrate Bdiabetogenic[ diet with rapamycin or placebo for 16 weeks. Rapamycin was administered admixed in the diet a microencapsulated form and
levels were detectable only in the drug-treated group.
Summary of Results: Mice that received rapamycin showed lesser weight
gain (pG0.01) compared to placebo fed animals despite equivalent food intake. Body composition analysis showed decreased body fat mass in the drug
treated group. Rapamycin-fed animals had lower plasma glucose (pG0.05)
and modest improvement in insulin sensitivity on insulin tolerance testing.
Plasma and hepatic triglyceride levels were reduced and morphologic analysis of the liver revealed decreased fat accumulation in hepatocytes in the
rapamycin group compared to the control animals. Immunohistochemical
analysis of perigonadal (visceral) adipose tissue showed decreased staining
for macrophage Mac2 in the mice treated with rapamycin. Gene expression
analysis of adipose tissue revealed decreased expression macrophage F4/80
and TNFa mRNA, while adiponectin gene expression was increased in the
rapamycin treated group.
Conclusions: Taken together, these data show that rapamycin improves the
metabolic profile and decreases adipose tissue inflammation in this mouse
model of diet-induced obesity. These results suggest that rapamycin and/or its
analogs have the potential for development as anti-obesity therapies.
111
PSYCHOLOGICAL INSULIN RESISTANCE IN A
LOW-INCOME, ETHNIC MINORITY POPULATION
Hsia SH, Machinani S, Bazargan-Hejazi S Charles R. Drew University,
Los Angeles, CA.
Purpose of Study: BPsychological insulin resistance[ is the unwillingness
of diabetic patients to begin insulin therapy, and substantially limits optimal
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
diabetes care. We examined the psychological factors underlying the
unwillingness to begin insulin therapy among low-income Hispanic and
African-American (AA) diabetic patients.
Methods Used: We conducted a cross-sectional survey of 156 non-insulintreated type 2 diabetic patients receiving care at a diabetes referral clinic in
inner city Los Angeles. The Survey for People Who Do Not Take Insulin
(SPI) was completed anonymously by consecutive patients during scheduled clinic visits from February to May 2010. Analyses were restricted to
insulin-naBve patients, and included the differential responses between Hispanic and AA patients, as well as correlations between demographic characteristics with the prevalence of unwillingness or the responses to each
survey item.
Summary of Results: A total of 136 respondents (87%) were insulin-naBve
(57% female; 69% Hispanic, 24% AA). Mean age and diabetes duration were
51.1 T 10.3 and 6.9 T 6.9 years, respectively; median monthly household
income and highest educational level categories were $200Y$1000 and grade
8Y12, respectively. Overall, 41.9% reported a complete unwillingness to
begin insulin. Compared to AA, Hispanic respondents were younger, lived
fewer years in the U.S., had less education, were significantly more likely to
be completely unwilling (46.8% vs. 26.5%, p=0.039), and reported a more
negative attitude to 8 of the 9 survey items (pe0.01 for each). Overall, fewer
years in the U.S. predicted greater unwillingness and a more negative attitude on 8 of the 9 survey items (pe0.03 for each); less education predicted
greater feelings of unfairness (p=0.01); Hispanic ethnicity predicted greater
feelings of failure (p=0.04); and males reported greater feelings of lifestyle
restriction (p=0.04).
Conclusions: Complete unwillingness to begin insulin use and negative
attitudes toward insulin are highly prevalent among low income, Hispanic
type 2 diabetic patients. These data may help to better guide culturally
appropriate patient counseling regarding insulin use.
112
DOES CALORIC INTAKE EXPLAIN THE WEIGHT-NEUTRAL
EFFECTS OF INSULIN DETEMIR VERSUS INSULIN
GLARGINE IN TYPE 1 DIABETES?
Garimella M, Mitchell S, Burge M University of New Mexico,
Albuquerque, NM.
Purpose of Study: Insulin detemir is a long-acting insulin analog that is
weight-neutral compared with other long-acting insulin formulations in
patients with type 1 diabetes. One mechanism for this observation may be an
effect of detemir on satiety. We hypothesized that type 1 diabetes patients
would eat fewer calories when presented with a standardized buffet meal
following a 24 hour fast when using insulin detemir as compared to insulin
glargine.
Methods Used: Ten subjects with C-peptide negative type 1 diabetes participated in a randomized, double blind crossover study in which they
received equivalent doses of either insulin detemir or glargine twice daily for
at least 3 weeks. They were subsequently admitted to the UNM Clinical
Research Unit for a 24 hour fast, after which they were allowed to eat to
satiety from a standardized, 10,000 calorie buffet. Content of the buffet was
based upon food preferences determined at screening, and all buffets were
identical in nutritional composition. Subsequently, all subjects repeated
the protocol with the other insulin. Meaurements of caloric consumption
following the fast, body composition by bioelectrical impedance, hunger
assessment, and indirect calorimetry were performed.
Summary of Results: Subjects were aged 35T11 years with a diabetes
duration of 18T11 years and HbA1c levels of 8T1%. There were 7 females
and 3 males, and BMI was 30T8 kg/ m2. Home capillary glucose control did
not differ during treatment with insulin detemir compared with insulin
glargine during the three week study (187T254 vs. 172T269 mg/dl; p=0.21),
but short acting insulin doses were higher on detemir (15T10 vs. 13T8 units;
pG0.001). Hunger scores did not differ between the study conditions following the 24 hour fast (68T25 vs. 73T24; p=0.58). Total energy ingested
from the buffet following the fast did not differ between insulin detemir
(1418T636 kcal) and insulin glargine (1357T576 kcal; p=0.63). Resting
energy expenditure also did not differ between detemir and glargine
(1405T398 vs. 1457T536 kcal/d; p=0.63).
Conclusions: The weight-neutrality of insulin detemir is not attributable to
reduced caloric intake in this controlled fasting study.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
113
EFFECT OF INSULIN ON SPILLOVER OF MEAL DERIVED
FATTY ACIDS
Nelson RH, Muthusamy K, Vlazny D, Smailovic A, Miles JM Mayo Clinic,
Rochester, MN.
Purpose of Study: Spillover, or release of chylomicron triglyceride fatty
acids by lipoprotein lipase (LPL) into the circulation as free fatty acids (FFA),
may contribute to the elevated total FFA seen in insulin resistant (IR) states.
A previous study in dogs suggested that spillover is regulated by intracellular lipolysis. The current study tested if exogenous insulin infusion during
meal absorption in IR adults, which produces greater FFA suppression than
the meal alone, results in reduced spillover.
Methods Used: Nondiabetic, IR adults (n=7) were studied on two occasions, a control day (C) and an insulin infusion day (I), using a nonrandomized design. Following a 5 day controlled diet and an overnight fast,
subjects were given a liquid meal made from a commercial nutrition supplement with additional vegetable oil to total 35% of calories as fat, 15% as
protein, & 50% as carbohydrate. Subjects sipped 1Y2 ounces of test drink
every 15 minutes for 6.5 hours to achieve steady state. A commercial lipid
emulsion labeled with [3H] triolein was infused to serve as a chylomicron
tracer, and 13C oleate was infused to measure the rate of appearance of FFA
and 3H oleate - the latter to allow calculation of spillover. The study was
repeated 2 weeks later in identical fashion except that the subjects received
insulin at 20 mU&mj2&minj1.
Summary of Results: Plasma FFA concentrations were higher on the C day
compared to I (124T13 v 75T8 Kmol/L, p G 0.01). Glucose concentrations
were not different during C compared to I (113T2 vs 113T1, p=NS). Glucose
infusion rate on I averaged 20T4 g/h. Plasma triglyceride concentration was
similar on C compared to I, both at baseline and during meal absorption
(181T23 v 168T29, p=0.50 & 257T22 v 230T35 mg/dL, p=0.12). Oleate rate
of appearance did not differ on C v I (341T27 v 297T53 HmoL/min, p=0.3).
Spillover was the same on C and I (28T3% vs 29T3%, p=0.77). Oleate
clearance was less with C than I (9T1 v 12T1 mL/min, p=0.02).
Conclusions: Our data indicate that suppression of intracellular lipolysis
with insulin does not reduce LPL-mediated spillover in humans during meal
absorption. Regulation of spillover may be related to noninsulin FFA suppression or other factors.
114
SITAGLIPTIN VERSUS THIAZOLIDINEDIONES AS A
THIRD-LINE ORAL ANTI-HYPERGLYCEMIC AGENT IN
TYPE 2 DIABETES MELLITUS
Hsia SH, Navar MD, Duran P, Shaheen M, Davidson MB Charles R. Drew
University of Medicine and Science, Los Angeles, CA.
Purpose of Study: To compare sitagliptin and thiazolidinediones (TZDs) as
a third-line oral anti-hyperglycemic agent among poorly controlled, ethnic
minority type 2 diabetic patients.
Methods Used: We treated 108 insulin-naive type 2 diabetic subjects who
were sub-optimally controlled on maximum tolerated doses of metformin
plus sulfonylureas with addition of sitagliptin 100 mg daily, and compared
their responses against a group of 104 similar patients treated with rosiglitazone (8 mg) or pioglitazone (45 mg) daily as their third-line oral agent,
using an identical treatment algorithm. Subjects were assessed bi-monthly,
and those who achieved HbA1c G 7.5% by 4 months were continued through
12 months of follow-up. Pioglitazone 45 mg daily was used as rescue therapy
for subjects who failed to achieve and/or maintain HbA1c G 7.5% at or
beyond 4 months.
Summary of Results: At baseline, sitagliptin- and TZD-treated subjects
had identical HbA1c (9.4 T 1.8% and 9.4 T 1.9%, respectively) and similar
known diabetes duration (6.7 T 5.0 and 7.6 T 5.8 years, respectively). HbA1c
was reduced in both groups at 4 months (pG0.001), but the reduction was
greater with TZDs than sitagliptin (j2.0 T 1.7% vs. j1.3 T 1.8%; p=0.006), as
was the proportion of subjects achieving HbA1c G 7.5% (61.5% vs. 46.1%;
p=0.026). Of all subjects achieving HbA1c G 7.5% at 4 months, the same
proportions of subjects in each group successfully sustained their HbA1c
G 7.5% by 12 months (59.1% vs. 57.8%). Of the sitagliptin subjects who
failed to achieve or maintain HbA1c G 7.5% and were given pioglitazone
45 mg daily as rescue therapy, 27.3% successfully achieved HbA1c G 7.5%
after an average of 4 months on pioglitazone. Sitagliptin was well tolerated,
with no significant weight gain, hypoglycemia, or other adverse events.
119
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Conclusions: Among ethnic minority type 2 diabetic patients very poorly
controlled on maximum tolerated doses of metformin and sulfonylureas,
third-line add-on therapy with a TZD controlled hyperglycemia more effectively than sitagliptin after 4 months. Our findings may have implications for
clinical practice recommendations.
&
Volume 59, Number 1, January 2011
factors for the development of insulin antibodies. Physicians need to include
insulin antibodies in the differential of precipitating causes of diabetic
ketoacidosis and be cautious about weaning intravenous insulin rapidly in
these patients.
115
INSULIN RESISTANCE IN ADOLESCENTS WITH TYPE I
DIABETES: ROLE OF HYPERGLYCEMIA
Pryor L1, Hull A1, Green MC1, Forster J4, Reusch JE3,2, Nadeau KJ1 1UC
Denver, Aurora, CO; 2UC Denver, Denver, CO; 3DVAMC, Denver, CO and
4
UC Denver, Aurora, CO.
Purpose of Study: Insulin resistance (IR) and associated metabolic syndrome (MetS) are likely responsible for the increased risk of cardiovascular
disease (CVD) in type 2 diabetes, but the etiology of accelerated CVD in type
1 diabetes (T1D) is unclear. We recently reported the paradoxical presence of
IR in lean T1D youth despite absence of the MetS. This IR may contribute to
CVD in T1D, but its mechanism is unknown. A potential cause of IR in T1D
may be hyperglycemia. A study reported IR in T1D adults that was unrelated
to HbA1c, but this study excluded marked hyperglycemia (HbA1c99.5
excluded, mean HbA1c 7.6%) and has not been replicated in youth. Thus, the
question of glucose_s role in the IR of T1D remains. We aimed to determine
if acute or chronic glycemia contributes to IR in T1D adolescents, a group
notorious for hyperglycemia.
Methods Used: We enrolled 66 lean, sedentary (G3 hours exercise/week),
T1D youth (44% male, age 15.5T2.2 years, HbA1c 8.6T1.5%, HbA1c range
6.2Y11.9%, BMI Z-score 0.38T0.96, %fat 24.9T7.5%) and 14 matched, lean,
sedentary nondiabetic controls. Following a 3-day control diet and overnight
insulin infusion, IR was determined by a 8o mu/m2/min hyperinsulinemic
euglycemic clamp.
Summary of Results: Fasting glucose prior to the clamp was
122.0T23.7 mg/dl in T1D youth and 83.9T5.1 mg/dl in control youth. T1D
youth were significantly more IR than controls (mean glucose infusion rate
8.5T3.3 vs. 15.3T4.7 mg/kg/min, pG0.01; 11.6T4.1 vs. 19.8T4.6 mg/lean.kg/min,
pG0.01) despite similar final glucose and insulin concentrations. By simple
linear regression, age, Tanner stage, BMI Z-score, and % fat mass, but not
gender, were significantly associated with IR in mg/kg/min among T1D
subjects (pG0.005, p=0.01, p=0.001, pG0.0001 respectively). While controlling for age, BMI Z-score, and percent fat mass, there was no significant
association between fasting glucose prior to the clamp or HbA1c, and IR
(expressed either per kg or per lean kg).
Conclusions: Similar to adults with lower HbA1c, T1D adolescents have IR
unrelated to acute or chronic glycemia or the MetS, suggesting the existence
of alternative unique mechanisms of IR in T1D. Future studies will aim to
detect these factors to target CVD risk-reducing therapies.
116
A UNIQUE CASE OF A 34 YEAR OLD WOMAN WITH
TYPE-2 DIABETES PRESENTING WITH DIABETIC
KETOACIDOSIS SECONDARY TO INSULIN ANTIBODIES
Garimella M, Schade DS University of New Mexico, Albuquerque, NM.
Case Report: High titer insulin antibodies are an unreported cause of
diabetic ketoacidosis. We describe the novel case of a 34 year old woman
with type 2 diabetes for 10 years (A1C=10.9), controlled on diet and oral
medications alone, who presented with severe diabetic ketoacidosis. Patient
noted that two weeks prior to admission she had a viral prodrome of fever and
rash along with unusually elevated blood glucoses in the 400s. This prompted
her to take 70/30 human insulin for 5 days. She had previously used 70/30
human insulin only during her two pregnancies, five and seven years prior
to admission. Initial insulin requirements on admission were 40 units/kg/day
(150 units/hour) intravenously for correction of the hyperglycemia and ketoacidosis. Further evaluation revealed an extremely high titer of insulin antibodies. When subcutaneous insulin was finally initiated on day 6, she relapsed
into diabetic ketoacidosis, indicating the continued presence of high titer
insulin antibodies. Her daily hospital chemistries and antibody levels are
shown in the table below.
Conclusion: The temporal relationship between the decline in elevated insulin antibodies and the improvement of diabetic ketoacidosis strongly suggest a causal relationship which has hitherto never been reported. Previous
intermittent use of parenteral insulin and viral illness are likely precipitating
120
Normal levels of insulin antibodies are G0.02 nmol/L
117
SYNERGISTIC EFFECT OF FRUCTOSE AND INSULIN ON
GLUCOSE METABOLISM IN HEPG2 CELLS
Patterson ME1,2, Wahjudi P2, Mao C1,2, Yee JK1,2, Lee PW1,2 1HarborUCLA Medical Center, Torrance, CA and 2LA BioMed, Torrance, CA.
Purpose of Study: The phenomenon that fructose is a better substrate for
glycogen synthesis than glucose is known as the glucose paradox. It has been
postulated that the increase in glycogen synthesis is the result of increases in
glucose uptake and gluconeogenesis in hepatocytes in the absence of insulin.
Whether the addition of insulin can further stimulate glycogen synthesis is
unknown. To test this possibility, we investigated the effect of fructose on
glycogen synthesis in HepG2 cells.
Methods Used: HepG2 cells were grown to confluence and incubated for
4 hours in medium containing 100 mg/dL glucose, 10 mM 50%U-13C3
lactate, and 1 mM sodium pyruvate. 4 sets of conditions were tested: no
fructose or insulin; with fructose, no insulin; no fructose, with insulin; and
with fructose and insulin. The medium concentrations of fructose and insulin
were 3 mM and 100 nM, respectively. Glucose uptake and cell glycogen
quantitation were carried out by GC/MS analysis with U-13C6 glucose as the
internal standard.
Summary of Results: Glucose uptake, which was slightly increased by
fructose and insulin compared with controls, was highest in the fructose plus
insulin group. Glycogen (Kg) had a similar trend with markedly increased
levels in the fructose (40.67 T 4.21) and insulin (18.43 T 3.5) groups compared with controls (10.7 T 1.22) with the largest increase in the fructose plus
insulin group (66.11 T 6.34). Glucose release (mg/dL) decreased with the
addition of insulin (9.65 T 1.09) and increased with fructose (13.9 T 0.65)
compared with glucose only (10.81 T 1.03). The group with fructose and
insulin showed a further decrease (8.64 T 0.61). Increased fatty acid synthesis
was seen with fructose (9.06 T 0.12%) and insulin (9.89 T 0.35%) compared
to the control group (7.76 T 0.11%) with the greatest increase in the fructose
plus insulin group (11.05 T 0.22%).
Conclusions: Our results show that the combined effect of insulin and
fructose increase glucose cycling and fatty acid synthesis, abnormalities
commonly found in metabolic syndrome and insulin resistant diabetes.
Whether these metabolic changes are the underlying mechanisms for the
association between high fructose consumption and the development of insulin resistance in metabolic syndrome remains to be investigated.
Neonatal Y Pulmonary I
Concurrent Session
8:30 AM
Friday, January 28, 2011
118
A NOVEL METHOD FOR THE DETECTION OF
NITRITE-DERIVED NITRIC OXIDE IN WHOLE BLOOD
Goltiao D1, Barceló LE1, Blood AB1,2, Power GG 2 1Loma Linda University,
Loma Linda, CA and 2Loma Linda University, Loma Linda, CA.
Purpose of Study: Nitric oxide (NO) is a potent vasodilator important to
blood flow regulation. Recent studies suggest that nitrite (NO2j), a compound found in plasma at È150 nM concentrations, can be converted into
NO by the following reaction with deoxyhemoglobin: deoxyHb + NO2j +
H+ Y metHb + NO + OHj.
Yet, as this reaction is more than a million times slower than the rates at
which NO itself is scavenged by reactions with hemoglobin, it is still unclear
whether this reaction can create vasodilating amounts of NO. Using a highly
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
sensitive method of measuring NO gas in liquid, this study aimed to measure
NO production from nitrite in whole blood. We hypothesized that nitrite
injections into blood at near-physiological concentrations would produce
vasodilating amounts of NO.
Methods Used: A closed 300-mL circuit with gas exchanger, heater, pump,
pressure and flow rate sensors, was filled with either phosphate buffered
saline (PBS) or with sheep blood circulating at 300 mL/min and deoxygenated by continuously perfusing the gas-phase side of the exchanger with
N2 and CO2. Gas exiting the exchanger was sampled for NO using a
chemiluminescence NO analyzer (NOA) sensitive to È5 parts per billion. The
sensitivity for detecting NO in the liquid phase was determined by injecting
free NO into circulating PBS. The production of NO from nitrite and
hemoglobin was measured by injecting sodium nitrite (50 to 500 KM) into
circulating blood. Methemoglobin was measured via spectrophotometry.
Summary of Results: Injections of NO into PBS resulted in linearly proportional (R2 = 0.995) detection of NO by the NOA from G10 nM to
910 KM concentrations. Although injection of nitrite into whole blood
resulted in the production of methemoglobin as predicted by the above equation, NO production in blood was not detected until nitrite concentrations
reached concentrations È1000 times higher than physiologic concentrations.
Conclusions: Using novel methods of detecting low nanomolar concentrations of NO in liquid, we were unable to detect NO production from nitrite
unless nitrite concentrations were È1000-fold higher than the physiological
range. These data suggest that the vasodilating properties of nitrite are derived
by mechanisms other than the reaction of nitrite with deoxyhemoglobin.
119
NITRITE INFUSION AT PHYSIOLOGIC CONCENTRATIONS
REDUCES CAROTID VASCULAR RESISTANCE IN
FETAL SHEEP
Truong G1, Schroeder H2, Bragg S1, Power GG2, Blood AB1,2 1Loma Linda
University Medical Center, Loma Linda, CA and 2Loma Linda University
Medical Center, Loma Linda, CA.
Purpose of Study: Nitrite, an anion present in mammalian plasma at midnanomolar concentrations, can be converted to the vasodilator nitric oxide
by reacting with deoxyhemoglobin. The reaction occurs faster with fetal
hemoglobin than adult, and may regulate fetal vascular tone under hypoxic
conditions. This experiment tests the hypothesis that nitrite influences fetal
carotid artery vascular resistance under physiological conditions.
Methods Used: We surgically instrumented fetal sheep for continuous
measurement of arterial blood pressure and carotid artery flow, from which
carotid resistance was calculated. After 4 to 5 days of recovery, ewes and
fetuses both received intravenous L-NNA to block nitric oxide synthase
activity. Nitrite was then infused into the fetal carotid artery (via a lingual
artery catheter) in doses increasing from 10 nM to 33,333 nM in 15 minute
steps. Samples for blood gases and nitrite concentrations were obtained
periodically throughout the experiment. One-way ANOVA was used to
determine significant changes over time.
Summary of Results: A significant increase in carotid resistance was observed after L-NNA infusion (pG0.01). Carotid artery resistance decreased
following initiation of nitrite infusion, reaching statistical significance during
the infusion of 333 nM nitrite (pG0.05).
Conclusions: We conclude that nitrite infusion to the carotid artery at nearphysiological concentrations significantly reduces carotid vascular resistance
to flow.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
120
PRENATAL ADMINISTRATION OF ROSIGLITAZONE TO
RAT PUPS DOES NOT ALTER THE ADULT METABOLIC
PHENOTYPE
Sierra H, Liu J, Sakurai R, Truong N, Lee RJ, Zhang G, Lee PW, Torday J,
Rehan V Harbor UCLA, Torrance, CA.
Purpose of Study: To study the metabolic profiles of adult rats exposed
prenatally to RGZ.
Methods Used: Pregnant Sprague-Dawley rat dams were administered either diluent, RGZ (0.3mg/kg), RGZ (3mg/kg), or Dexamethasone
(0.25mg/kg), intraperitoneally (IP) once daily 2 doses, 24h apart, starting
at embryonic day 18. At 11Y12 wks of age glucose and insulin tolerance
tests were performed after a 12h fast (glucose administered at 1g/kg IP,
and insulin administered at 1U/kg subcutaneously). De novo fatty acid
synthesis was analyzed by deuterium incorporation (DI) [99.9% deuterated
water (D2O)]. D2O was injected IP once at a dose equivalent to 4% of
body weight, followed by free access to drinking water containing 6%
D2O for 7 days] and mass spectrometry analysis at 14 wks. At 15 wks,
the animals were sacrificed and lungs, liver, skeletal muscle and fat
were collected for Western hybridization (WH) for PPARF (peroxisome
proliferator-activated receptor gamma) a marker of lung maturity, and
ADRP (adipose differentiation-related protein) a key down-stream functional target of PPARF. Plasma was assayed for triglycerides, cholesterol,
insulin, glucagon and troponin-I.
Summary of Results: Insulin and glucose tolerance tests were not different
between any groups. DI showed no significant differences in de novo fatty
acid synthesis, and WH showed equal expression of PPARF and ADRP.
Similarly, triglyceride, cholesterol, insulin, glucagon and troponin-I assays
revealed no significant differences among all 4 groups, (all p values 9 0.05).
Conclusions: Prenatal administration of RGZ does not alter the adult
metabolic phenotype. We speculate that RGZ therapy is a safe alternative to
dexamethasone for accelerating lung maturation and in preventing bronchopulmonary dysplasia in the permature infant.
121
LUNG LIPOFIBROBLASTS (LIFS) ACTIVELY RECRUIT,
STORE AND RELEASE RETINOIC ACID IN RESPONSE
TO PGE2
Truong N, Sakurai R, Rehan V, Torday J Harbor-UCLA, Torrance, CA.
Purpose of Study: Retinoic Acid (RA) is necessary for epithelial phenotypic expression and alveolar barrier function, yet treatment with exogenous
RA has not been found to consistently promote and protect the alveolar
epithelial barrier, either experimentally or clinically. We have previously
shown that the alveolar type II (ATII) cell increases PGE2 secretion when
stretched, stimulating Lipid Droplet (LD) secretion from neighboring lipofibroblasts (LIFs) at PGE2 concentrations comparable to those produced by
ATIIs in culture. Since RA is also stored in LIF LDs, we hypothesized
that PGE2 would concomitantly stimulate RA and lipid secretion by LIFs,
coordinating the release of ATII surfactant lipid substrate with RA, the latter
being an agonist for both surfactant phospholipid synthesis and surfactant
protein expression. Such a pleiotropic mechanism of RA action would
coordinate surfactant phospholipid and protein production with alveolar
distension, preventing atelectasis.
Methods Used: Uptake, storage and release of RA by cultured fibroblasts
was determined based on the kinetics of 3H-RA in cultured WI38 human
embryonic lung fibroblasts, primary fetal rat lung fibroblasts and adult lung
fibroblasts. Cells were treated with Prostaglandin E2 (5 10Y7M), an agonist
for the uptake and release of RA from LIFs. The sensitivity and specificity of
the PGE2 effects on LIF RA kinetics were determined using time-course
and dose-response curves in combination with the use of specific molecular
inhibitors (phloretin) and stimulators (butaprost) of the PGE2 receptor.
Summary of Results: LIFs actively recruit RA over a 1Y2 hour period, and
store it in lipid droplets. PGE2 (5 10Y7M) caused release of RA, whereas
baseline secretion was minimal.
Conclusions: LIF uptake and release of RA is coordinately regulated by
PGE2 signaling from ATII cells. This effect has novel implications for
understanding the basis for alveolar homeostasis and pathophysiology, providing new molecular targets for the treatment of Respiratory Distress
Syndrome, Bronchopulmonary Dysplasia, Interstitial Pulmonary Fibrosis
and Emphysema.(Partially supported by grants from the NIH (HL-55268,
121
Western Regional Meeting Abstracts
Journal of Investigative Medicine
HL-075405, HD058948, and HD-051857) and the TRDRP (14RT-0073,
15IT-0250, 17RT-0170).
122
PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR
(PPARF) AGONIST PROSTAGLANDIN J2 BLOCKS
HYPEROXIA-INDUCED DECREASE IN PULMONARY
FGF-10 SIGNALING
Sakurai R, Chen A, Torday J, Rehan V Los Angeles Biomedical Institute,
Torrance, CA.
Purpose of Study: Bronchopulmonary dysplasia (BPD) is characterized
by fewer and larger alveoli, and abnormal branching of the affected lungs.
Fibroblast growth factor 10 (FGF-10), an important growth factor that regulates lung branching and alveolarization, is decreased in BPD. On the other
hand, our lab has recently shown in a rat model that BPD can be prevented by
PPARF agonists such as prostaglandin J2 (PGJ2). How PGJ2 affects FGF-10
signaling is not known. The purpose of the study is to determine the effects of
FGF-10 on alveolar fibroblast differentiation and determine if PGJ2 blocks
hyperoxia-induced changes in FGF-10 signaling.
Methods Used: To determine the effects of FGF-10 on alveolar fibroblast
differentiation, both e19 (primary) fetal rat lung fibroblasts and human
embryonic (WI38 cells) were treated with FGF-10 (1-100 ng/ml) for 24h.
Following which, markers of fibroblast differentiation (PPARF, ADRP,
SREBP1c, aP2, leptin and triolein uptake) were analyzed. In separate experiments, neonatal Sprague Dawley rat pups were categorized in the following groups: normoxia (control, 21%O2), normoxia + PGJ2 (0.3mg/kg),
hyperoxia (exposure to 95%O2 for 7d), hyperoxia + PGJ2 (0.3mg/kg). PGJ2
was administered i.p. once daily. At the end of experimental period, the rat
pups were sacrificed and protein was extracted from whole lung samples.
Using Western blotting, we probed for FGF-10, and its receptor FGF receptor
2, PPARF, ADRP, leptin, fibronectin, Bax, Bcl-2 and cleaved caspase-3.
Summary of Results: FGF10-treated fibroblasts showed dose-dependent
increases in markers of differentiation. Under in vivo conditions, PGJ2
blocked hyperoxia-induced decreases in FGF-10, PPARF, leptin and increases in FGF-receptor 2 and fibronectin levels. Furthermore, hyperoxiainduced changes in apoptotic markers (increases in Bax and cleaved caspase
3 and decrease in Bcl-2) were also blocked in the PGJ2-treated group.
Conclusions: Our data demonstrate the down-regulation of FGF-10 signaling and increase in apoptosis on exposure to hyperoxia, both of which
were blocked by PGJ2. We speculate a direct interaction between FGF-10
and PPARF signaling pathways in the maintenance of alveolar growth
and differentiation (Grant support: NIH-HL55268, HL075405, HD058948,
HD051857) and TRDRP (15IT-0250, 17RT-0170).
123
URINARY NITRIC OXIDE METABOLITES (NOx) AND
CYCLIC GUANOSINE MONOPHOSPHATE (cGMP) IN
VENTILATED EXTREMELY LOW BIRTHWEIGHT (ELBW)
NEWBORNS RECEIVING INHALED NITIRIC OXIDE
Horneman H, Chapin C, Barrette A, Keller RL, Ballard R, Ballard P UC San
Francisco, San Francisco, CA.
Purpose of Study: ELBW newborns requiring mechanical ventilation after
7 d of age have 9 70% incidence of bronchopulmonary dysplasia (BPD). In
the NO CLD Trial, treatment with inhaled nitric oxide (iNO) increased survival without BPD (Ballard R, New Engl J Med 2006, 2007) and increased
levels of NOx in plasma and tracheal aspirate in a dose-dependent manner
(Posencheg J Perinatol 2009). The effect of infant iNO treatment on cGMP,
which mediates many responses to NO, is not known.
The objective of this study was to assess concentrations of NOx and
cGMP in urine of a new cohort of ELBW infants.
Methods Used: 379 urine samples were collected at intervals from 38
infants e 30 wk (mean 25.4 wk) in a pilot trial of late surfactant treatment. All
infants received 24 d of iNO per the NO CLD protocol. NOx was assayed
with a Siever_s NO Analyzer and cGMP was measured by Elisa (Cayman).
All data were normalized to urine creatinine (cre, Cayman) and expressed as
median values.
Summary of Results: Baseline values, defined as concentrations after
stopping iNO, were NOx 359 pmol/Kg cre and cGMP 8.3 nmol/Kg cre. NOx
increased in dose-dependent fashion with iNO exposure, plateauing between
10 and 20 ppm: 2 ppm 2.0-fold, 5 ppm 2.4-fold, 10 ppm 3.3-fold, and 20 ppm
122
&
Volume 59, Number 1, January 2011
2.9-fold (all e0.001). Corresponding values for cGMP were 1.1-fold (NS),
1.8-fold, 2.5-fold and 3.1-fold (each e0.002). The fold increase at 10 to
20 ppm was inversely correlated with baseline values for both NOx and
cGMP (r=j0.50/j0.48, p=0.004). Using mean infant values, urinary cGMP
was correlated with NOx at both baseline (r=0.43) and at iNO doses of
10Y20 ppm (r=0.35). Treatment with late surfactant did not affect either NOx
or cGMP.
Conclusions: We conclude that iNO treatment of ELBW infants increases
levels of both NOx and cGMP in the urine, reflecting delivery of NO to the
lung, stimulated cGMP production, and diffusion of both biomarkers into
plasma. Urinary NOx and cGMP are non-invasive measures of endogenous
and exogenous NO activity.
124
LIPOFIBROBLAST LIPID DROPLET TRAFFICKING
CO-REGULATES ALVEOLAR TYPE II CELL (ATII)
SURFACTANT PHOSPHOLIPID:PROTEIN EXPRESSION
Williams J, Sakurai R, Rehan V, Torday J Harbor-UCLA, Torrance, CA.
Purpose of Study: The mechanism coregulating surfactant phospholipid
and protein by ATIIs is largely unknown.Distension of lipofibroblasts (LIFs)
releases lipid droplets. We hypothesized that lipid droplets coordinately
stimulate surfactant phospholipid and protein.
Methods Used: Adipocyte differentiation-related protein (ADRP)-GFP
lipid droplets (LDs) normalized to their triglyceride content were incubated
with A549 cells. Subcellular localization of LDs was monitored by confocal microscopy. The effects of LDs on SP-B expression was determined by
PCR and Western analysis.LD stimulation of saturated phosphatidylcholine
(satPC) synthesis was determined by 3H-choline incorporation into satPC.
LDs were injected i.p. to mechanically ventilated rats to determine if ADRP
localized in the lung and stimulated surfactant synthesis.
Summary of Results: LDs localized in A549 nuclei within 10 mins,
appearing as prominent cytoplasm inclusions over the next two hours. Uptake
of the GFP-ADRP LDs was dose-dependent, increasing SP-B mRNA expression by 80% at 100 Kg/ml, and blocked by co-incubation with actinomycin D.LDs also increased SP-B protein expression in a dose-dependent
fashion, increasing SP-B by 70% at 100 Kg/ml, blocked by co-incubation
with cycloheximide. Incubation of A549s with LDs for 24h stimulated
satPC synthesis by 57-fold; co-incubation with ADRP antibody caused a
dosimetric decrease in LD-induced satPC synthesis. Uptake of LDs administered to mechanically ventilated adult rats (30 min) resulted in a dosedependent increase in lung ADRP content ranging from 5% at 45 Kg/kg to
20% at 4500 Kg/kg. SP-B protein increased between 5% at 45 Kg/kg and
20% at 4500 Kg/kg.
Conclusions: LDs rapidly translocate to ATII nuclei, stimulate SP-B
expression and surfactant phospholipid synthesis in an ADRP-dependent
manner. In vivo administration of LDs results in a dose-dependent increase
ADRP in lung, and a dosimetric increase in SP-B, consistent with the in vitro
findings. Therefore, stretch-regulated ATII-fibroblast interaction causes secretion of LDs that coordinately stimulate surface active phospholipid and
surfactant protein production. (Partially supported by grants from the NIH
(HL-55268, HL-075405, HD058948, and HD-051857) and the TRDRP
(14RT-0073, 15IT-0250, 17RT-0170)
125
LATE SURFACTANT INCREASES SURFACTANT PROTEIN
(SP) LEVELS IN VENTILATED EXTREMELY LOW
BIRTHWEIGHT (ELBW) NEWBORNS
Keller RL1, Merrill J2, Palermo L3, Truog W4, Steinhorn R5, Ryan R6,
Eichenwald E7, Durand D2, Courtney S8, Black D3, Ballard P1, Ballard R1
1
UCSF, San Francisco, CA; 2CHORI, Oakland, CA; 3UCSF, San Francisco,
CA; 4Mercy Children’s Hospital, Kansas City, MO; 5Children’s Memorial
Hospital, Chicago, IL; 6WCHOB, Buffalo, NY; 7UT Houston, Houston, TX
and 8SBUMC, Stony Brook, NY.
Purpose of Study: ELBW newborns requiring mechanical ventilation after
7 d of age have 9 70% incidence of bronchopulmonary dysplasia (BPD), and
most infants experience episodes of surfactant dysfunction secondary to SP-B
deficiency (Merrill, Peds Res 2004). Treatment with inhaled nitric oxide (INO)
increases survival without BPD (Ballard R, NEJM 2006,2007) but does not
abolish surfactant dysfunction. We investigated the safety and efficacy of late
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
doses of surfactant (calfactant) plus INO to improve surfactant composition
in ventilated preterm infants.
Methods Used: Randomized, blinded trial of ELBW (e 1000g and 30
0/7 wk GA) ventilated at 73Y14 d. Infants received INO plus 5 dose procedures of surfactant (SURF) or sham (CTRL). SP-B content (% of total
protein) was quantified by immunoblot assay in large aggregate surfactant
isolated from tracheal tracheal aspirate samples. 85 infants were enrolled;
48 were treated every 3Y4 d (SURF n=28 and CTRL n=20) and the remainder were treated weekly after the first 2 doses.
Summary of Results: Infants treated every 3Y4 d that received SURF
(vs. CTRL) were less mature (25.0 vs. 25.7 wks, P=0.05), and more likely
male (68 vs. 40%, P=0.06). SP-B content at study entry was lower in SURF
(1.06%) vs. CTRL (2.34%, P=0.05). Compared to baseline levels, SP-B
content increased 0.69% (P=0.0003) and 0.05% (P=0.11) on days 1 and 2
after SURF; content decreased slightly in CTRL. The change in SP-B content on d 1 for treated infants was correlated with the mean levels of SP-B
content (r=0.60, p=0.004), perhaps indicating enhanced SP-B degradation
in deficient infants. There was no difference (SURF vs. CTRL) in PDA
ligation (41 vs. 28%, P=0.37), NEC (11 vs. 15%) or IVH 3/4 (4 vs. 5%) after
enrollment. 201 doses of surfactant were administered with adverse event
frequency e 2% (reintubation, or significant bradycardia or respiratory
decompensation).
Conclusions: Late surfactant increased SP-B content. Administration of
late surfactant is well-tolerated and appears safe with regard to co-morbidities
of prematurity.
126
CALCIUM/CALMODULIN-DEPENDENT PROTEIN
KINASE 2 AS A POTENTIAL MEDIATOR OF
BETAMETHASONE-INDUCED FETAL LUNG MATURATION
Nielson JJ, Li C, Li A, Chan B, Ramanathan R, Minoo P Keck School of
Medicine, University of Southern California, Los Angeles, CA.
Purpose of Study: Antenatal Steroids are in common clinical use for
promoting maturation and hence improving perinatal outcome in preterm
neonates. The precise mechanism by which steroids enhance lung development & improve respiratory functions remains unknown. Calcium/calmodulindependent protein kinase (CaMK2) may be an important mediator of
corticosteroid-induced fetal lung maturation. The current study examined the
relationship between the maturational impact of betamethasone on fetal lung
and activation of CaMK2 in a murine model.
Methods Used: The pattern of CaMK2 activation (phosphorylation) was
examined during lung development by western blot analysis. Antenatal
betamethasone was administered to pregnant mice on day 14 (E14) of gestation. The course of the regimen was chosen to resemble what is used in
humans. Two doses of betamethasone separated by 24 hours were administered. Saline was used as control. Fetuses at E16 were delivered by hysterotomy and the lungs isolated for characterization. The lungs were examined
for gross morphology and expression of a number of pulmonary cell differentiation makers including Clara cell-specific kD 10 protein, beta tubulin,
alpha smooth muscle actin, surfactant proteins B and C, and T1a. Activation of CaMK2 was conducted by detection of Phospho-CaMK2 by western
blot analysis.
Summary of Results: CaMK2 activation correlated with the onset of saccular stage during lung development. Betamethasone treatment was associated
with enhanced fetal lung maturation, cell differentiation and concomitant activation of CaMK2.
Conclusions: To our knowledge, these are the first data demonstrating a
close association between CaMK2 activation and lung structural maturation.
The results also show CaMK2 as a target of betamethasone, a well known
stimulator of lung maturation in clinical use. We speculate that CaMK2 may
be an important mediator of betamethasone-induced lung maturation.
127
THIRDHAND SMOKE ADVERSELY AFFECTS FETAL LUNG
GROWTH AND DIFFERENTIATION
Chien K, Sakurai R, Torday J, Rehan V Los Angeles Biomedical Institute,
Torrance, CA.
Purpose of Study: Though nicotine is the main cigarette smoke constituent linked to altered lung structure and function in in utero smoke ex-
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
posed infants, other constituents have not been thoroughly evaluated. This
is particularly true for thirdhand smoke (THS), defined as tobacco smoke that
sorbs onto indoor surfaces and dust, where semi- and non-volatile chemicals
and particulates can undergo modifications to produce new toxicants that remain on the surfaces, or later desorb and re-appear in the micro-environment.
We tested the hypothesis that THS exposure alters the growth and differentiation of the developing lung by affecting the alveolar PPARF signaling known
to be essential for normal lung development.
Methods Used: Embyronic day 19 fetal rat lung explants were exposed
to diluent, nicotine (110Y8M or 110Y5M), or 4-(methylnitrosamino)-1(3-pyridyl)-1-butanone (NNK) (110Y8M or 110Y5M), the main tobaccospecific N-nitrosamine constituent of THS, for 24h. Cell proliferation (BrDU
incorporation), cytotoxicity (LDH assay), and expression of key markers of
alveolar differentiation (surfactant protein B and C and PPARF expression,
choline incorporation into saturated phosphatidylcholine and triolein uptake)
were determined.
Summary of Results: Cell proliferation decreased and cytotoxicity increased significantly with NNK compared to nicotine (pG0.05), suggesting
more adverse effects of NNK. With NNK, choline incorporation into saturated phosphatidylcholine decreased significantly (pG0.05), while it increased
significantly with nicotine. Triolein uptake increased significantly with both
nicotine and NNK (pG0.05), though more robustly with NNK (pG0.05 vs.
nicotine). Western analysis showed significantly decreased PPARF and increased fibronectin levels (pG0.05) with both NNK and nicotine exposure.
Conclusions: NNK affects lung growth and differentiation more robustly
than nicotine, possibly contributing significantly to in utero smoke exposureinduced pulmonary damage. These data can be used in designing specific
preventive and/or therapeutic strategies against the deleterious effects of in
utero THS exposure on lung development [Supported partially by grants
from the TRDRP (15IT-0250, 17RT-0170).
128
MECHANCAL VENTILATION OF PRETERM LAMBS LEADS
TO MORE APOPTOSIS AND LESS PROLIFERATION OF
NEURONS AND GLIA
Alvord J, Wint A, McCoy MJ, Dong L, Dahl M, Callaway C, McKnight R,
Yoder BA, DiGernimo R, Lane RH, Albertine K University of Utah, Salt
Lake City, UT.
Purpose of Study: Prolonged mechanical ventilation (MV) of the premature human neonate is associated with lung injury. Frequently, the brain also
is injured. Our studies, using chronically ventilated preterm lambs, indicate
that MV changes histone marks in the brain. These changes could affect
regulation of cell processes, such as apoptosis and proliferation of neurons
and glia. The present studies purpose was to test this possibility.We hypothesized that prolonged MV increases apoptosis and decreases proliferation
among neurons and glia in the brain of preterm lambs.
Methods Used: Preterm (PT) lambs, treated with antenatal steroids and
postnatal surfactant,were managed by MVor high-frequency nasal ventilation
(HFNV; akin to bubble nasal CPAP (n=4 each) for either 3d or 21d. We use
HFNV as the positive gold-standard for alveolar formation in the lung. At
the end of 3d or 21d of ventilation, cortical brain tissue from the temporal
lobe was fixed and analyzed by double immunofluorescence to colocalize
cleaved caspase 3 or PCNA within neurons (neuronal marker-positive), immature oligodendrocytes (O1-positive), or reactive astrocytes (GFAP-positive).
Summary of Results: Neurons in gray matter had significantly (*pG0.05)
more colocalization of cleaved caspase 3 in the MV group compared to the
HFNV group at 3d (22T3* vs 7T3/mm3) and at 21d (22T3* vs 11T4).
Conversely, neurons had significantly less colocalization of PCNA in the MV
group compared to the HFNV group at 3d (17T3* vs 23T7) and at 21d (27T7*
vs 16T1). Among glial cell types, the only colocalization difference was for
immature oligodendrocytes with cleaved caspase 3 at 21d. Significantly
more for immature oligodendrocytes were apoptotic in the MV group compared to the HFNV group (29T3* vs 16T1).
Conclusions: We conclude that MV for 3d or 21d is associated with a shift
in balance between cell death and proliferation in the brain. The shift is
toward more apoptosis and less proliferation of neurons and immature oligodendrocytes. These results are consistent with less brain-derived neurotrophic factor in the same MV preterm lambs compared to We speculate
that imbalance results from epigenetic alterations in chromatin structure
of genes that regulate apoptosis and proliferation. (HL62875, HL56401,
HD41075).
123
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Neonatology Y General I
Concurrent Session
8:00 AM
Friday, January 28, 2011
&
Volume 59, Number 1, January 2011
Conclusions: We conclude SCG contributes to regulation of CBF in our
model. Our study suggests stimulating the SCG exerts localized vasoconstriction of CBF. Better understanding of the vasoconstrictor effect of the
SCG in regulation of CBF may lead to effective treatment of premature babies
at risk for IVH.
129
TRACT BASED SPATIAL STATISTICS OF NEONATAL
HYPOXIC-ISCHEMIC ENCEPHALOPATHY: SELECTIVE
INVOLVEMENT OF CORTICAL SPINAL TRACT
Weiner A1,2, Ceschin R 3, Wisnowski J 4, Tokar E 5, Nagasunder A 4, Bluml S 4,
Ramanathan R1,2, Paquette L 2,1, Panigrahy A3,4 1USC, Los Angeles, CA;
2
CHLA, Los Angeles, CA; 3Children’s Hospital of Pittsburgh of UPMC,
Pittsburgh, PA; 4CHLA, Los Angeles, CA and 5USC, Los Angeles, CA.
Purpose of Study: HIE in the newborn is heterogeneous with respect to
both neuroimaging patterns and etiologies. In this study, diffusion tensor
imaging (DTI) combined with a TBSS-based analysis were used to test the
hypothesis that selective white matter (WM) tracts in the newborn brain are
vulnerable to HIE despite the pattern of injury seen on conventional MRI
or etiology.
Methods Used: 12 neonates with HIE were scanned between 2005Y2009
using a standardized neonatal imaging protocol and a neonatal head coil on
a 1.5T GE magnet. The inclusion criteria included having a pattern of central and/or peripheral HIE on conventional and diffusion sequences. A group
of term clinical controls with normal conventional MRs was used for comparison. The DTI protocol included 25 directions at a b-value=700. Data
analysis was performed using Oxford University_s FMRIB FSL software.
Voxel-wise statistics were performed using Threshold-Free Cluster Enhancement and corrected for multiple comparisons. Diffusivity metrics were
calculated using a ROI approach and DTI studio. Clinical variables were
recorded.
Summary of Results: The etiologies for HIE in this group were heterogeneous and included perinatal depression, traumatic birth injury, metabolic
disease, non-accidental trauma, and hypotension. In the HIE group, the one
minute Apgar ranged from 1Y3, the five minute Apgar from 0Y5 and the
pH of the first ABG ranged from 6.81Y6.86. TBSS revealed in the HIE cases
selective reduction of anisotropy of the posterior limb of the internal capsule (PLIC) when compared to term controls (pG0.05). No selective patterns
related to the diffusivity metrics were found.
Conclusions: TBSS reveals selective microstructural WM injury in the
PLIC in neonates with different neuroimaging patterns and etiologies of
HIE, suggesting a common vulnerability of the cortical spinal tract fibers.
This could be related to the high metabolism of these structures at birth as
documented by other imaging modalities including PET.
130
ELECTRICAL FIELD STIMULATION OF THE SUPERIOR
CERVICAL GANGLION ATTENUATES CEREBRAL BLOOD
FLOW IN TERM AND PRETERM LAMBS
Czynski AJ1, Blood AB1, Buchholz J2 1Loma Linda Univ., Loma Linda, CA
and 2Loma Linda Univ., Loma Linda, CA.
Purpose of Study: The role of the superior cervical ganglion (SCG) in
controlling cerebral autoregulation is controversial. Most studies involve
adult human or animal models. There are no reports describing SCG function in autoregulation of preterm infants. We hypothesis that SCG regulates
cerebral blood flow (CBF) in premature lambs.
Methods Used: Premature lambs 120Y124 d (n=6) and 2-week-old term
lambs (n=5) were anesthetized, mechanically ventilated, and instrumented.
Arterial PCO2 and PaO2 were maintained in a constant range. CBF was
monitored using laser Doppler flow (LDF) technique. After placement of
LDF probes, the left or right SCG was randomly selected, and platinum
electrodes were placed. Following a stable baseline of CBF, the SCG was
electrically stimulated for 30 seconds 3Y6 times with a return to baseline
between stimulations. Distribution of values for each CBF time point were
analyzed using Friedman 2 Way ANOVA. Differences at specific time points
compared to baseline were determined by Wilcoxon Signed Rank test.
Summary of Results: With SCG stimulation CBF decreased (p=0.01) on
the ipsilateral side, but was unchanged on the contralateral side, and no
change in BP. After stimulation stopped CBF increased but remained lower
than baseline (p=0.018).
124
131
COMPARISON OF UMBILICAL VENOUS AND
INTRAOSSEOUS ACCESS DURING SIMULATED
NEONATAL RESUSCITATION
Rajani A, Chitkara R, Halamek L Stanford University School of Medicine,
Palo Alto, CA.
Purpose of Study: Current Neonatal Resuscitation Program guidelines
recommend placement of an umbilical venous catheter (UVC) for any neonate with persistent bradycardia in spite of appropriate cardiopulmonary resuscitation or hypovolemic shock. UVC placement is complex, imposes
significant space constraints on the resuscitation team, and is only required
in 0.12% of all births. Successful intraosseous needle (ION) placement and
use in neonates is well described, and animal models suggest that epinephrine administered intraosseously has similar effects to other intravenous
routes. This study compares UVC and ION placement in the delivery room
setting to understand time to placement, ease of use and errors in placement
for each device.
Methods Used: Forty physicians were recruited (8 attending neonatologists,
5 neonatal hospitalists, 6 fellows in neonatology, 5 neonatal nurse practitioners, 16 residents in pediatrics). Participants were shown an instructional
video of UVC and ION placement and were allowed to practice placement
with both modalities. Using a realistic delivery room and neonatal mannequin
in a pediatric simulation center with a confederate nurse and respiratory
therapist, participants were asked to attend the two deliveries requiring intravenous epinephrine: one scenario utilized UVC catheterization and the
other ION placement. Subjects were randomly assigned to perform the procedures either in the UVC-IO or IO-UVC order. The scenarios were recorded
using video cameras and the tape was reviewed for time to placement and
errors. Participants were then surveyed regarding the level of difficulty of
each device.
Summary of Results: ION placement was significantly faster than UVC
placement by 46 seconds (pG0.0001). Overall, there was no difference in
perceived difficulty between IO and UVC placement. Residents in pediatrics
(n=16) found ION placement to be significantly easier than UVC (pG0.003).
There was no significant difference in the number of errors between UVC
and IO placement.
Conclusions: ION placement can be performed more quickly than UVC
without any increase in overall perceived difficulty or rates of error. ION
insertion should be considered as a first line option for epinephrine administration in the delivery room, especially for practitioners who do not
routinely place UVCs in the intensive care setting.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
132
COMPARISON OF CARDIAC OUTPUT MEASUREMENTS
BETWEEN ELECTRICAL CARDIOMETRY (EC) AND
ECHOCARDIOGRAPHY
Cayabyab RG1, Bhombal S2, Ebrahimi M1, Seri I 1,2 1LAC+USC
Medical Center, Los Angeles, CA and 2Childrens Hospital Los Angeles,
Los Angeles, CA.
Purpose of Study: To investigate the correlation of cardiac output measurements obtained by the non-invasive continuous cardiac output monitor
(electrical cardiometry, Aesculon\) and echocardiography in term and preterm infants.
Methods Used: This is a prospective observational study. Left ventricular
output (LVO) was measured by echocardiography and EC simultaneously. As
EC estimates LVO by assessing blood flow in the ascending aorta, measurements were performed on term and preterm infants with a patent ductus
arteriosus irrespective of the potential hemodynamic significance of ductal
shunting. Measurements were obtained from day 1Y72 of postnatal life.
Summary of Results: There were 46, paired measurements performed on
neonates with a gestational age of 23Y41 and birth weight of 375Y4330gms.
LVO by ECHO and EC was 653T188, and 658T263 mL/min, respectively
(r2=0.55).
Conclusions: Despite the relative imprecision of LVO measurement by
echocardiography, there is a good correlation between the cardiac output
measurements obtained by EC and echocardiography in neonates. There is a
potential for continuous monitoring of cardiac output with the use of EC in
these infants but additional studies simultaneously investigating changes in
CO in response to treatment are needed to confirm these findings.
133
TRACT BASED SPATIAL STATISTICS (TBBS) OF
ECMO-RELATED NEONATAL BRAIN INJURY:
INITIAL EXPERIENCE
Arora V1, Ceschin R2, Wisnowski J3, Nagasunder A3, Bluml S3,
Ramanathan R1, Paquette L1, Panigrahy A3,2 1Childrens Hospital Los
Angeles, Los Angeles, CA; 2Children’s Hospital of Pittsburgh of UPMC,
Pittsburgh, PA and 3Childrens Hospital of Los Angeles, Los Angeles, CA.
Purpose of Study: Neonates treated with ECMO (extra-corporeal membrane oxygenation) are at risk for long- term neurodevelopmental sequelae.
The precise mechanism of brain injury is not yet understood. In this study,
we used diffusion tensor imaging (DTI) combined with a TBSS-based analysis
to test the hypothesis that ECMO results in selective microstructural injury
involving white matter (WM) in the developing neonatal brain.
Methods Used: 28 neonates who had been on ECMO were scanned between 2005Y09 using a standardized neonatal imaging protocol and neonatal
head coil on a 1.5T GE magnet. There were 18 veno-venous ECMO patients
and 10 veno-arterial. The reason for ECMO included primary PPHN (n=6),
cardiac arrest (n=2), sepsis (n=5), congenital diaphragmatic hernia (n=8),
meconium aspiration syndrome (n=6) and dilated cardiomyopathy (n=1).
Due to technical limitations associated with scan acquisition and/or postprocessing of the data, a total of 11 ECMO cases were used for the analysis.
The comparison groups without ECMO included: (a) term and preterm clinical
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
controls with normal conventional MRIs (n= 60), (b) congenital heart disease
(CHD) (n=45). The DTI protocol included 25 directions at a b-value=700. Data
analysis was performed using Oxford University_s FMRIB FSL software.
Voxel-wise statistics were performed using Threshold-Free Cluster Enhancement and corrected for multiple comparisons. Clinical variables were recorded.
Summary of Results: TBSS revealed no difference in fractional anisotropy of selected WM tracts in the ECMO cases compared to both preterm
and term controls. When comparing ECMO cases to the CHD cases, there
was decreased anisotropy in the splenium of the corpus callosum (CC) of
CHD patients.
Conclusions: Our preliminary results show no definite anisotropic differences between ECMO neonates and clinical controls. This suggests that
ECMO does not increase the risk for brain microstructural injury. In contrast,
CHD patients did demonstrate decreased anisotropy in the splenium of
the CC compared to ECMO patients, which is likely a neurodevelopmental
aberration in CHD neonates.
134
THE EFFECT OF ANTITHROMBIN III ON BLOOD
PRODUCT REQUIREMENTS DURING NEONATAL
EXTRACORPOREAL MEMBRANE OXYGENATION SUPPORT
Perry R1, Klee L1, Ramanathan R1, Stein J2, Seri I1, Friedlich P1 1USC
Division of Neonatal Medicine, Childrens Hospital Los Angeles, USC Keck
School of Medicine, Los Angeles, CA and 2USC Division of Pediatric
Surgery, Childrens Hospital Los Angeles, USC Keck School of Medicine,
Los Angeles, CA.
Purpose of Study: Neonates receiving extra corporeal membrane oxygenation (ECMO) support frequently require transfusion of blood product.
Due to their relative immaturity and their underlying medical and surgical
conditions, neonates treated with ECMO are often deficient in Anti-thrombin
III (ATIII). This in turn further enhances their propensity for blood product
exposures during ECMO support. The purpose of this study is to determine
whether ATIII administration alters the need for blood product administration during ECMO support.
Methods Used: Neonates requiring ECMO at the NICCU of Childrens
Hospital Los Angeles were divided in a pre-ATIII (years 2006Y2008) and
post-ATIII (years 2009Y2010) administration periods. One vial of ATIII is
administered to patients for ATIII level typically less than 60%. Patient demographics, length of ECMO support and blood product [fresh frozen
plasma (FFP), packed red blood cells (PRBC), cryoprecipitate and platelets]
requirements were compared between the two time periods.
Summary of Results: In this ongoing study, so far five patients in the
pre-ATIII periods (Mean birth weight = 3290 +/j 223 g, gestational age =
38 +/j 1 weeks, length of ECMO support = 205 +/j 79 hours) and five
patients in the post-ATIII periods (Mean birth weight = 3295 +/j 307 g, gestational age = 38 +/j 1 weeks, length of EMCO support = 234 +/j124 hours)
were identified. During ECMO, patients in the post-ATIII period were exposed to significantly less FFP as compared to the pre-ATIII period (mean
FFP volume = 48 +/j 11 vs. 126 +/j 44 ml; p = 0.04). No differences in the
volume exposure of PRBC, cryoprecipitate or platelets were noted between
the two groups.
Conclusions: Our preliminary data suggest that supplementation of ATIII
during neonatal ECMO support may result in a reduction of FFP exposure.
135
DEVELOPMENTAL MASSAGE THERAPY (DMT) PROMOTES
PARASYMPATHETIC NERVOUS SYSTEM ACTIVITY IN
PRETERM INFANTS
Smith S1, Haley S2, Wong B1, Moyer-Mileur LJ 2 1Univ of Utah, Salt Lake
City, UT and 2Univ of Utah, Salt Lake City, UT.
Purpose of Study: Preterm infants are exposed to numerous challenges that
elicit a stress response. Massage therapy is reported to decrease stress biomarkers, however, the effect of developmental massage therapy (DMT) on
autonomic nervous system (ANS) activity in preterm infants has not been
tested. We hypothesized that DMT would improve ANS function by promoting parasympathetic activity concomitant with decreased sympathetic
activity.
Methods Used: Preterm, 28Y32 wk infants were randomized to DMT or
Control. DMT infants received a 20-min DMT session by a licensed massage therapist twice daily for 4 wks. Control infants received no stimulation
125
Western Regional Meeting Abstracts
Journal of Investigative Medicine
as the therapist stood at bedside. Masking to treatment was maintained with
a privacy screen. Weekly heart rate variability (HRV) data, from which highfrequency power (parasympathetic) and low-frequency power (sympathetic)
were extracted, was acquired prior to the morning treatment session. The lowto-high frequency ratio (LF:HF) assessed ANS function with a lower ratio
indicative of improved parasympathetic activity.
Summary of Results: 36 infants (DMT n=16/7M & Control n=20/10M)
were studied. Infant demographic and anthropometric characteristics were
similar between groups. DMT infants demonstrated significantly greater
parasympathetic activity over time (F = 5.47, p = 0.03) (Figure 1).
Conclusions: Parasympathetic response is essential for recovery following
stress-induced sympathetic activity. We demonstrate twice-daily DMT promotes greater parasympathetic activity in preterm infants. DMT may be a
useful therapy for attenuation of stress in hospitalized, preterm infants.
136
A RANDOMIZED, MASKED, PLACEBO CONTROLLED
STUDY TO ASSESS THE SAFETY AND EFFICACY
OF DARBEPOETIN ALFA ADMINISTERED TO
PRETERM INFANTS
Ohls R1, Christensen RD3, Wiedmeier S3, Kamath BD2, Rosenberg A2,
Schrader R1, Backstrom-Lacy C1, Lambert D3, Burnett J3, Pruckler B2,
Peceny H1 1UNM, ABQ, NM; 2UC, Denver, CO and 3IHC, SLC, UT.
Purpose of Study: A novel erythropoiesis stimulating agent (ESA),
darbepoetin alfa (Darbe) increases hematocrit (Hct) in adults with end stage
renal disease or cancer, dosing every 1Y3 weeks. Weekly Darbe administration has not been evaluated in preterm infants. We hypothesized that
infants would respond to Darbe in similar fashion to erythropoietin (Epo),
by increasing erythropoiesis and decreasing transfusion need. We measured
erythropoietic response and number of transfusions in preterm infants randomized to weekly Darbe, thrice weekly Epo, or placebo (sham dosing).
Methods Used: Infants e1,250 grams and e48 hours of age were randomized
in masked fashion to Darbe (10 Kg/kg, 1/wk SC), Epo (400 units/kg, 3/wk
SC) or placebo. Dosing continued to 35 weeks gestation. All infants received
supplemental iron, folate, and vitamin E. Complete blood counts, absolute reticulocyte counts (ARC), transfusions, phlebotomy losses, and adverse events
were recorded. Infants were transfused according to a standardized transfusion
protocol.
Summary of Results: A total of 102 infants (946T196 grams, 27.7T
1.8 weeks gestation, 51T25 hours of age) were enrolled. There were no
significant differences between treatment groups at baseline. Infants in the
Darbe and Epo (ESA) groups required fewer transfusions (p=0.01) and were
exposed to fewer donors (p=0.02) than the placebo group (Epo: 1.2 transfusions and 0.8 donors/pt; Darbe: 1.4 transfusions and 0.6 donors/pt; placebo:
2.6 transfusions and 1.4 donors/pt). Hct and ARC were higher at the end of
study in the ESA groups compared to placebo (pG0.001, ESA versus placebo
for both Hct and ARC). Hospital morbidities were similar between groups,
126
&
Volume 59, Number 1, January 2011
including the incidence of ROP, and were no different from previously
published incidences in very low birth weight infants, despite strict transfusion guidelines.
Conclusions: Infants receiving ESAs required fewer transfusions and were
exposed to fewer donors without adverse effects or increased morbidities.
The application of restrictive transfusion guidelines did not increase morbidities among preterm infants. Long term neurodevelopmental follow-up is
in progress (NCT# 00334737; supported by Thrasher Research Fund).
137
EFFECT OF FLUORESCENT LIGHT EXPOSURE ON
METALLOPORHYRIN-TREATED NEWBORN MICE
Schulz-Geske S, Kalish FS, Zhao H, Katayama Y, Champion KA,
Vreman HJ, Wong RJ, Stevenson DK Stanford University School of
Medicine, Stanford, CA.
Purpose of Study: Neonatal hyperbilirubinemia is due to an increased
bilirubin (BR) production and a decreased capacity to eliminate BR. Blue
light (400Y520 nm) is commonly used to reduce BR levels. However, a more
strategic approach might be to prevent BR production. Metalloporphyrins
(Mps) effectively decrease BR formation by competitive inhibition of heme
oxygenase (HO), the rate-limiting enzyme in heme degradation. Chromium
mesoporphyrin (CrMP) and zinc deuteroporphyrin bis glycol (ZnBG) are
promising Mps for use since they are absorbed orally, highly potent, and affect
minimally other hemoproteins. Because some Mps are photosensitizers, the
phototoxicity of CrMP and ZnBG were studied in a 3-d-old mouse model.
Methods Used: 3-d-old FVB mice were given vehicle (CON), CrMP,
or ZnBG [3.75Y30.0 Kmol/kg body weight (BW) IP]. For light treatment,
pups were placed in an open box lined with wet paper towels, set under
2 cool white and 1 blue fluorescent (TL52) tubes at an irradiance of
35.0T1.0 KW/cm2/nm for 3h (CON-; CrMP-; ZnBG-L), and wetted every
30min with water. Age-matched treated pups were returned to mothers and
kept under ambient light (CON, CrMP, ZnBG). Kaplan-Meier plots were
used to determine survival (n=10/group). Liver tissue HO activity, antioxidant capacity, and plasma markers of liver and cardiac muscle injury
(AST, CK-MB) were measured in each group.
Summary of Results: ZnBG-L and CrMP-L treatment resulted in a dosedependent mortality with an LD50 of 20.0 and 21.5 Kmol/kg BW, respectively. However, in contrast to ZnBG, there was no significance difference in
mortality between CrMP-L and CrMP groups. ZnBG-L pups had significant
weight loss (1.7T1.0%), decreased liver antioxidant capacity (4.7-fold, pe0.05),
and increased AST (1.4-fold, pe0.05) and CK-MB (10.0-fold, pe0.05) levels
compared to CON. Moreover, the inhibitory potency of ZnBG for HO decreased significantly by È15% after light exposure. In summary, CrMP is not
phototoxic, but shows chemical toxicity with an LD50 of 22.5 Kmol/kg BW;
whereas, ZnBG is potentially phototoxic.
Conclusions: We conclude that at low doses (G3.75 Kmol/kg BW), ZnBG
has a high inhibitory potency, but low phototoxicity similar to tin mesoporphyrin (SnMP), and thus may still have potential use in the treatment of
neonatal hyperbilirubinemia.
138
ROLE OF HEME OXYGENASE IN A MURINE MODEL OF
EARLY NECROTIZING ENTEROCOLITIS
Schulz-Geske S1, Kalish FS1, Jang KY2, Zhao H1, Huey M1, Vreman HJ1,
Sylvester KS2, Wong RJ1, Stevenson DK1 1Stanford University School of
Medicine, Stanford, CA and 2Stanford University School of Medicine,
Stanford, CA.
Purpose of Study: Necrotizing enterocolitis (NEC) is typified by mucosal
and intestinal necrosis. Three Bstress[ factors are usually present: a preceding
ischemic event, bacterial colonization, and enteral feeding, causing increased
intestinal permeability and injury followed by exaggerated inflammatory
response. Heme oxygenase-1 (HO-1) degrades heme to bilirubin (BR), free
iron, and carbon monoxide (CO). Since BR and CO have antioxidant, antiapoptotic, and anti-inflammatory properties, we propose that HO-1 may protect against the development of NEC.
Methods Used: 10d-old HO-1 heterozygote (Het, HO-1+/j) and WT mice
were used. For NEC induction, all pups were removed from the mom, orally
gavaged with 200KL formula (2g Similac 60/40 in 10mL Esbilac)/5g BW q
4h x 2d and exposed to hypoxia (5%O2 x 2min before feeding, 2x/d, n=5).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
At d3, pups were returned to moms. Control pups (WT-; Het-CON) remained
with moms and breastfed. At d4, all pups were sacrificed and $1.0cm of
distal ileum (IL) and proximal colon (C) were harvested for morphology.
To compare NEC severity, tissues were sectioned for H&E staining. Histological changes were blindly scored on a scale of 0 (normal) to 4 (severe
inflammation/necrosis). Apoptosis in IL and C of CON and NEC-induced
mice were assessed by TUNEL staining.
Summary of Results: In this pilot study, histological scoring showed
morphological changes of IL and C for Het-CON vs WT-CON. NEC development was more advanced and severe in Het-NEC vs WT-NEC pups
(Table). Interestingly, tissue damage was higher in the C than the IL for both
NEC-induced genotypes. TUNEL staining showed an increase in apoptosis
in Het-CON IL and C vs those of WT-CON. Moreover, apoptotic cells
were strongly reduced in Het-NEC C vs WT-CON, Het-CON, and WT-NEC,
which may be a sign of reduced proliferation and the presence of necrotic
tissue.
Conclusions: A partial deficiency in HO-1 leads to an earlier, more progressive and severe development of experimental NEC. Also, it appears that
the early development of NEC may originate in the proximal colon in this
murine model.
139
MAGNETIC RESONANCE IMAGING (MRI) TO ASSESS
CEREBRAL INJURY AND PREDICT
NEURODEVELOPMENTAL OUTCOMES IN INFANTS BORN
WITH CONGENITAL DIAPHRAGMATIC HERNIA (CDH)
Chen M, Bonifacio SL, Glass HC, Rogers EE, Barkovich J, Keller RL UCSF,
San Francisco, CA.
Purpose of Study: Children with CDH are at risk for adverse neurodevelopmental (ND) outcomes. However, there is little systematic data on
the MRI findings of brain injury that precede these adverse outcomes. The
purpose of this study was to determine the pattern of MRI abnormalities in
infants with CDH and the association of these findings with ND outcomes.
Methods Used: We performed a retrospective cohort study of 99 infants
with CDH cared for from birth at UCSF (1998Y2009) who underwent ND
evaluation. Postnatal MRI in 24 newborns over the same period were classified as Normal (n=7) or Abnormal (n=17) based on the determination of
cerebral injury by a pediatric neuroradiologist. ND outcome (n=22) was
classified from the Bayley Scales of Infant Development (II or III). Bayley
scores were used to categorize ND outcome into Normal, Borderline
(1Y2 SDs below the mean) and Abnormal (92 SDs below the mean). Statistical analysis was performed with Rank Sum or Chi Square Test.
Summary of Results: 24/24 infants who underwent MR imaging needed
non-primary repair, as compared to 36/75 infants who did not undergo imaging, P=G0.01. Imaged infants required a greater median number of days
before successful extubation (27 days) compared to infants who were not
imaged (11 days), P=G0.01. Among those with abnormal MRI, the predominant pattern demonstrated was white matter injury (WMI) (14/17).
Children with WMI were less likely to have normal ND outcome (5/13)
compared to those with normal MRI (4/6), P=0.25. Children with WMI also
had lower median MDI and PDI scores compared to children with normal
MRI (MDI 71 vs. 86, P=0.46; PDI 62 vs. 86, P=0.09), although the comparisons did not reach statistical significance.
Conclusions: The majority of MRI studies in this cohort demonstrated
cerebral injury, although more severely affected infants were more likely
to undergo imaging. WMI was the predominant manifestation of injury, the
same pattern identified in association with prematurity and congenital heart
disease. Infants with abnormal MRI are at increased risk for adverse ND
outcome. Thus, MR imaging in infancy may be useful for identifying children with CDH at increased risk for adverse ND outcome.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
140
RISK FACTORS FOR EPILEPSY IN TERM INFANTS WITH
HYPOXIC ISCHEMIC ENCEPHALOPATHY
Hong KJ1, Bonifacio SL3, Barkovich AJ4, Sullivan JE2,3, Rogers EE3,
Ferriero DM2,3, Glass HC2,3 1University of Washington, Seattle, WA;
2
University of California San Francisco, San Francisco, CA; 3University of
California San Francisco, San Francisco, CA and 4University of California
San Francisco, San Francisco, CA.
Purpose of Study: Hypoxic ischemic encephalopathy (HIE) is a significant cause of neonatal death and adverse neurodevelopmental outcome, such
as cerebral palsy, developmental delay, and epilepsy. The goal of this study
was to determine neonatal predictors of epilepsy in term newborns with HIE
using clinical data and MRI.
Methods Used: This was a cohort study of 195 term infants with birth
asphyxia, born 1993 through 2009. Neonatal clinical data, such as neonatal
seizures, encephalopathy scores, and MRI pattern of injury, were obtained
through chart review. Parents of the infants were contacted by telephone and
administered a seizure questionnaire. Neurodevelopmental data including
cognitive scores, neuromotor scores (NMS), and neurological outcomes were
extracted from standardized longitudinal follow up data.
Summary of Results: Of 121 children contacted, 12 (10%) developed
epilepsy. Univariate analyses showed significant differences between epilepsy and no epilepsy groups for neonatal seizures, EEG seizures, status
epilepticus, encephalopathy scores of 5 or 6, and brain injury on MRI
(PG0.01). Of 49 infants with neonatal seizures, 12 (24%) developed epilepsy. The infants with EEG seizures (RR=9; 95% CI=3Y27) and status
epilepticus (RR=11; 95% CI=5Y25) were more likely to develop epilepsy
than those without. There was no significant association for clinical seizures
(P=0.5) or encephalopathy scores of 3 or 4 (P=0.1) and epilepsy. The children with epilepsy had injury on MRI: 5 had basal ganglia patterns and 7
had watershed patterns. Children with epilepsy had an average cognitive
score of 54T11, whereas children with no epilepsy had a score of 97T17
(PG0.005). Median (range) NMS were 5 (0Y5) for children with epilepsy
and 0 (0Y5) for children with no epilepsy (PG0.005).
Conclusions: Term infants with HIE and neonatal seizures, specifically
electrographic seizures and status epilepticus, evidence of brain injury on
MRI, and more severe encephalopathy scores are at a higher risk of developing epilepsy. Our data provide information for clinicians to use when
counseling parents on the possible outcome of epilepsy.
141
COMPARISON OF CEREBRAL REGIONAL OXYGEN
SATURATION (CRSO2) AND RENAL REGIONAL OXYGEN
SATURATION (RRSO2) RESPONSE TO OXYGEN
DESATURATION IN EXTREMELY LOW BIRTH WEIGHT
INFANTS (ELBW) DURING THE FIRST 72 HOURS
OF POSTNATAL LIFE
Soleymani S1,2, Cayabyab R1,3, Seri I1,3 1Children Hospital, Los Angeles,
CA; 2University of Southern California, Los Angeles, CA and 3University of
Southern California, Los Angeles, CA.
Purpose of Study: To study the difference in cerebral and renal response
to oxygen desaturations in ELBW during the first 72 hours of postnatal life.
Methods Used: Preterm infants 23Y28 weeks_ gestation were continuously
monitored using the INVOS cerebral/somatic oximeter (Somanetics, Troy,
Michigan), on the forehead and the flank respectively, during the first
72 hours of postnatal life. The output of the INVOS and other hemodynamic
parameters including SaO2, heart rate and blood pressure were simultaneously collected via the analogue output of the patient monitor on to a laptop
computer. In areas of data which showed SaO2 desaturation from baseline,
percent change for SaO2, CrSO2 an RrSO2 were calculated (Figure1).
Summary of Results: Data for 7 patients with a gestational age of 25.9T
1.7 weeks and birth weight of 899T 152g were used for the analysis. Mean
and standard deviation for percent change in SaO2, RrSO2, CrSO2 and
difference between RrSO2 and CrSO2 were calculated (Table1).
Conclusions: This data shows that renal response to oxygen desaturation
is statistically different compared to the cerebral response (pG.0001). These
findings indicate that even very preterm neonates are capable to respond to
decreases in O2 delivery with non-vital organ (renal) vasoconstriction to
maintain appropriate O2 delivery to the brain under adverse conditions.
127
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Percent Change in Oxygen Saturation
143
TEMPORAL ALTERATIONS IN LESION VOLUME IN A RAT
MODEL OF REPETITIVE MILD TRAUMATIC BRAIN INJURY
Data is presented as mean(std).
Neuroscience I
Concurrent Session
9:00 AM
Friday, January 28, 2011
142
AMYLOID BETA INJECTION IN AXONAL TERMINAL SITES
RESULTS IN MOUSE OPTIC TRACT DEGENERATION
Carrick D1, Sun S2,3 1Loma Linda University, Loma Linda, CA; 2Loma
Linda University, Loma Linda, CA and 3Loma Linda University, Loma
Linda, CA.
Purpose of Study: Amyloid plaques are a pathological hallmark of
Alzheimer_s disease (AD) and the principal component of these plaques is
amyloid beta (AA). Although AA has been implicated in AD pathogenesis, the
mechanism of AA-induced neurodegeneration is not clear. In this study, we
applied novel magnetic resonance diffusion tensor imaging (DTI) to identify
white matter damage as a result of focal exposure of axonal terminals to AA.
Methods Used: In the mouse visual system, the optic nerve and the optic
tract are extended axons originating from the retinal ganglion cells (RGC).
Therefore, the axonal terminal sites are located in the optic radiation in the
subcortical brain areas while the neuronal bodies of the RGC sit in the eye.
AA peptides were injected into the right hemisphere optic radiation region of
six 12 week old female C57BL/6 mice. DTI was conducted on each mouse at
1 and 3 months after AA injection.
Summary of Results: Among the measured white matter tracts, only optic
tracts and optic nerves showed AA effects. The ipsilateral optic tract showed
a significant 15% decrease in axial diffusivity, suggesting axonal damage
caused by AA injection. Damage in the optic tract propagated further to the
optic nerves, which showed significant 10% decreases of mean diffusivity.
Histological examination will be performed to confirm the imaging findings.
Conclusions: This study demonstrated induced axonal degeneration in response to axonal terminal site exposure to AA peptides.
128
Muellner M1, Coats J2, Mohd-Yusof A4, Neglario K3, Obenaus A2,5,6,
Huang L6 1Loma Linda University, Loma Linda, CA; 2Loma Linda
University, Loma Linda University, CA; 3University of California,
Riverside, Riverside, CA; 4California State University, San Bernardino,
San Bernardino, CA; 5Loma Linda University, Loma Linda, CA and 6Loma
Linda University, Loma Linda, CA.
Purpose of Study: Repetitive mild traumatic brain injury (rmTBI) is
an important medical concern for active military personnel with 10Y20%
reporting neurological or psychological symptoms. In the present study, we
characterized the neuropathological profiles of repetitive mTBI using a rat
model where the first mild impact was then followed by a second mTBI
at intervals of 1,3, or 7 days by non-invasive magnetic resonance imaging
(MRI) correlated with histology.
Methods Used: Thirty-four Sprague Dawley adult male rats (2 mo old)
were randomized into sham, single mTBI, and 3 groups with two episodes
of mTBI: 1) 1 day apart; 2) 3 days apart; 3) 7 days apart. A craniotomy
was performed followed by a mild controlled cortical impact (CCI) delivered
by an electromagnetic driven piston (0.5 mm depth, 4 mm diameter tip at
6.0 m/s, 200 ms duration). A second identical impact was delivered at 1, 3
or 7 days after the first CCI event at the same location. Sham animals underwent the same surgical procedure without CCI. T2 weighted imaging
(T2WI) was acquired on a 4.7T MRI (Bruker Biospin) at 24 hrs after each
impact and a final MRI was acquired at day 14 after the initial injury. Results
were correlated with ex vivo histology.
Summary of Results: In the rmTBI groups of 1 and 3 days but not 7 days
apart, a second CCI resulted in increased T2WI lesion volumes that persisted
until 14 days (pG0.05 vs sham).
Conclusions: The brain appears to exhibit heightened vulnerability to a
second mild traumatic insult up to 3 days after an initial mTBI event. Rat
models of rmTBI may serve as a clinically relevant platform for evaluation
of outcome parameters for testing experimental therapeutics. MRI is a sensitive neuroimaging biomarker for monitoring the pathological evolution
after rmTBI.
144
ELEVATED LEVELS OF CASPASE 14 IN PATIENTS WITH
ALZHEIMER_S DISEASE
Van Ornam J1, Crofton A1, Schrag M1, Zabel M1, Mueller C2, Vinters H3,
Kirsch W1 1Loma Linda University, Loma Linda, CA; 2George Mason
University, Fairfax, VA and 3UCLA Medical Center, Los Angeles, CA.
Purpose of Study: Caspase 14 is a recently discovered aspartate protease
that was found to be elevated in a patient who progressed from control to
mild cognitive impairment and who subsequently progressed to and died of
Alzheimer_s disease (AD). The purpose of this current study is to elucidate
the significance of caspase 14 in the etiology of Alzheimer’s disease.
Methods Used: This study utilized flash-frozen brain samples from patients
grouped (n=4 in each group) into control, AD-only, and AD with cerebral
amyloid angiopathy (CAA) via post-mortem pathological diagnosis. Standard immunoblotting was used to compare the levels of caspase 14 in the
brain parenchyma of each group and in brain parenchyma versus isolated
brain vasculature. Standard immunohistochemistry was also performed on
brain sections from each group in order to localize changes in caspase 14.
Human polyclonal caspase 14 antibody was used for both techniques.
Summary of Results: LC-MS/MS demonstrated a 25-fold increase of
caspase 14 from serum in a patient progressing from control to MCI. Immunoblotting of brain homogenate demonstrated a statistically significant
increase (p=0.004) in caspase 14 in AD-only parenchyma compared to
control and AD+CAA parenchyma. Immunoblotting of brain microvessels
showed a strong non-significant trend towards increased caspase 14 expression in both AD-only and AD+CAA compared to control. Immunohistochemistry of AD-only brain sections were negative for intraneuronal staining
of caspase 14 while control and AD+CAA brain sections stained positive.
The vasculature of AD-only and AD+CAA brain sections was also prominently stained.
Conclusions: Our results suggest that AD+CAA cases follow a different
progression of disease than AD-only cases. Future studies will aim to clarify
the role of caspase 14 in Alzheimer_s disease.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Purpose of Study: The percentage of prescription drug abusers in the
145
SURGERY DURATION VERSUS BRAIN IN JURY IN THE
NEONATAL RAT MODEL OF HYPOXIA-ISCHEMIA
Burris M, Chen H, Fajilan A, Zhang JH Loma Linda University School of
Medicine, Loma Linda, CA..
Purpose of Study: In humans, hypoxia-ischemia causes brain injury which
may lead to neurological deficits. For study of neonatal hypoxia-ischemia,
preclinical models have been developed to allow testing of neuroprotective
agents. The purpose of this study is to evaluate the relationship between the
surgical duration and the severity of brain injury in the neonatal rat.
Methods Used: Unilateral carotid ligation was performed in postnatal
day 7 and 10 rat pups, followed by 2.5 hours of enclosed-chamber hypoxia
(8% oxygen). Time-to-ligation of each study group was set at 5, 7, 13 and
21 minutes.
Summary of Results: There was a statistically significant difference between the groups of surgical durations of 5 and 7 minutes compared with
those of 13 and 21 minutes.
Conclusions: The operative time is a major determinant in the resulting
cerebral infarction following unilateral carotid ligation and incubation in an
hypoxic chamber in P7 and P10 rat pups.
Native American Population increased more than six-fold between 1998Y2008
(Treatment Episode Data Set, July 2010). This, compared to the four-fold increase seen in the general population, demonstrates the increasing concern
of narcotic addiction among Native Americans. In recent months, this issue
was brought to the attention of health care providers working on the Nez-Perce
Indian Reservation in Lapwai, ID, when it became evident that a number of
pain patients had become addicted to their prescribed narcotics. Under the
Controlled Substances Act, it is illegal for a provider to continue to prescribe
an addictive substance to an identified addict. The purpose of this project was
to increase awareness and facilitate understanding of the most recent scientific
evidence explaining the biological model of addiction.
Methods Used: To assist providers with understanding the mechanism
of addiction, a review of the most recent addiction research was conducted.
A 30-minute presentation summarizing the role of narcotics in the body
and Dr. Volkow_s recent research describing the predisposition for addiction based on genetically predetermined levels of dopamine receptors in the
frontal lobe was presented to the entire clinic (physicians, nurse practitioners,
physician_s assistants, nurses, medical assistants, behavioral health specialists,
and community health workers). Additionally, copies of the presentation were
made available to the staff for future reference.
Summary of Results: The focus of this presentation was directed toward
health care providers to encourage a deeper understanding of how a patient
unknowingly becomes a prescription drug addict. The latest scientific evidence
has demonstrated a genetic predisposition for addiction- a condition previously
explained through social and psychological stressors. Approximately 50 providers attended the presentation, which encouraged further discussion of narcotic treatment plans.
Conclusions: There are many social theories attempting to demystify why
Native American populations suffer from a disproportionate percentage
of substance abuse. Examined here, is the latest scientific research demonstrating the role of genetics and dopamine receptor levels in prescription
narcotic addiction.
147
INHIBITION OF ASTROCYTE GLT-1 IN THE
VENTROMEDIAL HYPOTHALAMUS ENHANCES THE
EPINEPHRINE RESPONSE IN HYPOGLYCEMIA
COUNTERREGULATORY FAILURE
P7 Surgery Time vs Infarction Percentage
P10 Surgery Time vs Infarction Percentage
146
INVESTIGATING THE CURRENT BIOLOGICAL MODEL
OF ADDICTION
Lawhorn JK University of Washington School of Medicine, Seattle, WA.
* 2011 The American Federation for Medical Research
Shalygin N1,2, Sanders N2 1University of Washington School of Medicine,
Seattle, WA and 2VA Puget Sound Health Care System, Seattle, WA.
Purpose of Study: Hypoglycemia is a major limiting factor in the
achievement of tight glycemic control with intensive insulin therapy. With
repeated exposure to hypoglycemia, CNS-mediated activation of hormonal
counterregulatory responses (CRRs) that normally correct low glucose become
impaired (Hypoglycemia-Associated Autonomic failure, HAAF). Glutamate
signaling within the VMH is essential to the activation of hypoglycemiainduced CRRs. Thus, changes in how glutamate is regulated should be a
mechanism to consider in the development of HAAF. We hypothesize that
hypoglycemia-induced up-regulation in astrocyte GLT-1, a glutamate re-uptake
transporter responsible for clearing extracellular glutamate and terminating
signaling, may be a contributing factor in the development of CRR failure.
Elevated GLT-1 expression would be expected to reduce the availability of
glutamate, thereby impairing hypothalamic glutamate mechanisms mediating
the activation of CRRs.
Methods Used: To test this hypothesis, we injected a rodent model with
dihydrokainic acid (DHK; 1KM in 0.5 Kl/side), a selective inhibitor of astrocyte GLT-1, bilaterally into the VMH and measured glucose and hormonal
CRRs to single (SH) and recurrent (RH) hypoglycemia.
Summary of Results: In response to SH, epinephrine (EPI) levels were
significantly elevated in response to DHK treatment (2,031+239) as compared to ACSF controls (1,134+152) (t.60; pG0.05). The recurrent hypoglycemia (RH) model, as expected, dramatically reduced EPI levels as
compared to SH controls (630+72 vs. 1,134+152, t.60; p=0.024). Injection
of DHK into the VMH prior to the final bout of RH effectively restored
the impaired EPI response to levels observed in SH controls (1,280+194 vs.
1,134+152, t.60; p=0.027). Selective inhibition of GLT-1, in the VMH glucosesensing site enhanced a key CRR to hypoglycemia, adrenal medullary epinephrine release, and prevented the typical downregulation in this CRR that
occurs with repeated hypoglycemia.
129
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Conclusions: These findings suggest that hypoglycemia-induced upregulation in GLT-1 expression and/or function may impair glutamatergic
signaling required for the full activation of hormonal CRRs and reveal a
potentially important and novel astrocyte-mediated mechanism in the development of HAAF.
148
MEASURING DEPRESSION SEVERITY IN PATIENTS
WITH EPILEPSY
Dehdashti H1, Labiner D2, Chong J3 1University of Arizona College of
Medicine, Tucson, AZ; 2University of Arizona College of Medicine, Tucson,
AZ and 3University of Arizona College of Medicine, Tucson, AZ.
Purpose of Study: The purpose of the study was to identify the different
factors which contribute to depression in patients with epilepsy and to compare
relevant findings with patients who only have epilepsy.
Methods Used: Questionnaire packets were given to patients at local
neurology clinics. These packets contained questions relating to the patient_s
Quality of Life (QOL). Patients were asked to rate their QOL and social
support scores. A statistical software package tabulated the data and determined different QOL scores from the patient surveys. The scaled responses
of the patients provided quantitative data which was plotted and analyzed.
Summary of Results: Patients with both epilepsy and depression scored
higher in perceived criticism, stigmatization, and lower self-efficacy. Compared to patients with epilepsy only, these patients were criticized more by
family and friends, felt a greater sense of stigma in the community, and
experienced much lower self-esteem. In another aspect of QOL, patients with
epilepsy and depression also had lower social support scores, less appraisal,
sense of belonging, stigma and tangible levels. They received less treatment
for their condition, lacked familial support and social interaction, felt stigmatized in society, and had little access to transportation and financial aid.
However, families of patients from both groups showed approximately the
same level of emotional support.
Conclusions: Our data showed that patients with both epilepsy and depression experience a much lower QOL compared to patients with only epilepsy.
This shows that depression serves as a comorbidity for patients with epilepsy.
Our methods for acquiring patient data helped illustrate this point by using
variables and scores to determine the QOL in these two different patient groups.
149
STUDY OF THE NEUROPHYSIOLOGY OF COGNITIVE
DYSFUNCTION IN PARKINSON’S DISEASE
Baker KA University of Colorado Denver, Aurora, CO.
Purpose of Study: Cognitive dysfunction affects 20Y40% of persons with
Parkinson_s disease (PD) at the time of diagnosis and is a significant risk
factor for psychosis, dementia, nursing home placement and death. Among
individuals with PD surviving 20 years or longer, cognitive dysfunction is
the leading cause of nursing home placement and 75% eventually develop
dementia. These statistics have not significantly changed since the 1920_s
despite major advances in the treatment of motor symptoms. This study
builds on recent advances in cognitive neuroscience and an innovative neurophysiologic research technique to address this glaring gap in our knowledge and treatment of PD.
Methods Used: We will use magnetoencephalography (MEG) to investigate cognitive dysfunction in subjects with PD performing a demanding
cognitive task (modified cued Stroop paradigm). For each trial, subjects will
receive an instructional cue, either BColor[ or BWord[, indicating whether
they are to name the color or read the word of the upcoming stimulus. The
stimuli are color words written in colored font. The stimuli may be congruent
(e.g. Bred[ written in red letters) or incongruent (e.g. Bred[ written in blue
letters). Subjects will perform this task continuously for 30 minutes while in
their on-medication state. We plan to collect data on 10-20 PD subjects and
10Y20 age-matched controls.
Summary of Results: Preliminary evidence shows that the Stroop paradigm is an effective method of determining cognitive dysfunction mediated
by executive control, indicating that this study is capable of demonstrating
cognitive differences between PD and non-PD populations.
Conclusions: We hypothesize that top-down networks, particularly those
related to medial frontal structures affected by PD, are critical in mediating
cognitive dysfunction. We predict that cognitive dysfunction will correlate
with medial frontal brain activity.
130
150
REPRODUCIBILITY OF GAMA-BAND RESPONSE
RECORDED BY MAGNETOENCEPHALOGRAPHY AND
ELECTROENCEPHALOGRAPHY
Simon ST, Rojas DC University of Colorado Health Sciences Center,
Denver, CO.
Purpose of Study: Patients with schizophrenia often exhibit unusual sensory experiences, ranging from visual or auditory distortions to dynamic hallucinations. Oscillatory neuronal electrical activity in the range of 30Y50 Hz
(the gamma-band oscillation) has been proposed to involve feature binding or
inter-regional communication within the brain and is critically dependent on
inhibitory neurotransmission within the cerebral cortex. Gamma-band oscillatory power has been reported to be impaired in persons with schizophrenia
and is also found in the first-degree relatives of persons with schizophrenia,
suggesting a heritable component. Magnetoencephalography (MEG) and
electroencephalography (EEG) technologies have repeatedly shown this effect,
making the measure an exciting potential tool that could be useful to other
researchers. However, no one has yet performed test-retest measurements in
the same group of subjects. This study is a necessity before such data can be
used in large-scale projects such as genetic linkage analyses or as biomarkers
in clinical trials.
Methods Used: Twenty healthy and normal hearing adults screened for
personal and family history of mental illness and neurological disorders will
be recruited. The subjects will be recorded in an auditory steady state response (ASSR) paradigm while having a 64-channel EEG recording followed
by a 248-channel MEG recording. One week later, they will return and repeat
the same procedures. We will be comparing two stimuli in the MEG and
EEG runs - amplitude modulated white noise and click trains, binaurally presented at 75 dB SPL. Both produce strong 40 Hz responses. We will be looking
at phase-locked response amplitude and phase-locking factors, in both sensor
and source space. The reliability of the source localization will also be a secondary interest variable. Analysis of ASSR-derived measures of gamma-band
power and inter-trial phase-locking for retest reliability will ensue.
Summary of Results: With five of twenty subjects completed, no results
have yet been analyzed.
Conclusions: We expect to see a high level of test-retest reliability with
the MEG and expect it to be significantly greater than that seen in the EEG
recording.
151
THE EFFECT OF PRENATAL ALCOHOL EXPOSURE ON
SUBSEQUENT ETHANOL CONSUMPTION AFTER THE
ADOLESCENT EXPERIENCE OF A TRAUMATIC EVENT
IN MALE C57BL/6J MICE: A PILOT STUDY
Drexler KJ, Caldwell KK University of New Mexico School of Medicine,
Albuquerque, NM.
Purpose of Study: Fetal Alcohol Spectrum Disorder (FASD) affects
0.2Y1% of the population (Sampson et al. 1997; US Department of Health
and Human Services 2005). People with this disorder may demonstrate differential behavioral responses to stressful situations (Kodituwakku 2007,
Hellmans 2010). As part of a series of studies on the interaction of stress
and alcohol abuse, we aimed to test the hypothesis that prenatal exposure
to ethanol predisposes individuals to the development of alcohol misuse
following trauma exposure.
Methods Used: Female C57BL/6J mice were allowed to binge drink a
10% ethanol solution for the duration of pregnancy. Six male offspring from
prenatal alcohol exposure (PAE) litters and six male offspring from saccharin
control litters were provided access to 20% ethanol solutions according to
the Drinking In the Dark model described by Rhodes et al (2005) beginning
at age 45Y50 days. Binge drinking before and after two cycles of withdrawal were recorded. The third withdrawal cycle was coupled with exposure
of the animals to a traumatic event via predator odor exposure after which
binge drinking was again assessed.
Summary of Results: Both PAE and control animals demonstrated a strong
alcohol deprivation effect following the first withdrawal but not following
the second withdrawal. Alcohol consumption increased in both groups following the traumatic exposure with the PAE animals trending toward a greater
increase in drinking after trauma than the controls; however, this difference did
not reach statistical significance (p = 0.1635). Trauma-exposed mice maintained drinking levels, which is not seen after withdrawal cycles prior to trauma.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
There was a significant difference between the maintenance of drinking levels
following the first, second and third withdrawal (with trauma) (pG0.0001) but,
again, the PAE and control groups did not significantly differ from each other.
Conclusions: Exposure to a traumatic event during adolescence increases
binge drinking in both control and PAE C57BL/6J male mice. PAE mice
show a heightened sensitivity to trauma, as evidenced by a greater increase
in drinking which may reach statistical significance with a higher powered
study.
152
MGMT BIOCHEMICAL ACTIVITY IS ASSOCIATED
WITH MYELOSUPPRESSION FOLLOWING THERAPY
WITH TEMOZOLOMIDE
Olson J, Silber J, Chamberlain M University of Washington, Mercer
Island, WA.
Purpose of Study: Inclusion of the methylating agent temozolomide
(TMZ) during radiation therapy, then continuing TMZ as a single agent
after radiation can significantly prolong patient life. However, treatment with
TMZ may result in clinically significant leukopenia, neutropenia, anemia
or thrombocytopenia in a minority of patients. Suppression of bone marrow
function threatens the efficacy of TMZ therapy by necessitating dose reduction or discontinuance. Such clinical limitations emphasize the need to develop
methods to identify patients susceptible to TMZ-induced myelotoxicity. The
purpose of this study is to examine the association between clinically relevant
myelosupression and expression of O6-methylguanine-DNA methyltransferase
(MGMT), the DNA repair activity that removes cytotoxic O6-methylguanine
(O6-meG) DNA adducts induced by TMZ, in human peripheral blood leukocytes (PBLs).
Methods Used: To test our hypothesis, we assayed MGMT biochemical
activity and determined MGMT promoter CpG methylation status, a surrogate
measure of gene expression, in three patient populations (n=28); 10 glioma
patients treated with TMZ showing no myelotoxicity; 8 glioma patients treated
with TMZ showing myelotoxicity (defined as clinically relevant Grade 3 or
higher myelotoxicity that resulted TMZ drug reduction, delay in therapy or
discontinuance) and 10 disease-free, untreated controls. MGMT promoter
CpG methylation status determined by methylation-specific PCR and MGMT
activity was measured by standard biochemical assay that assays transfer of
O6-[3H]meG from DNA to protein.
Summary of Results: Mean MGMT activity was 2-fold lower in patients
with myelotoxicity compared to treated patients with normal counts (8.3 T
3.9 vs 16.3 T 7.8 fmol/106 cells; P e 0.014) and to disease-free, untreated
controls (8.3 T 3.9 vs 15.7 T 6.8 fmol/106 cells; P e 0.011). All samples
displayed unmethylated promoters.
Conclusions: These data suggest that [1] myelotoxicity in TMZ-treated
patients reflects a reduced capacity to remove TMZ-induced O6-meG, [2]
promoter CpG methylation status is not indicative of MGMT expression
in PBLs and [3] low MGMT activity be a marker for increased risk for
TMZ-induced myelotoxicity.
Pulmonary and Critical Care I
Concurrent Session
8:30 AM
Friday, January 28, 2011
153
DIFFUSE OPTICAL SPECTROSCOPY MONITORING OF
CYTOCHROME C OXIDASE REDOX STATE DURING
PHYSIOLOGICAL CHALLENGES IN ANIMAL MODELS
Lee J1, Kim JG1, Mahon S1, Mukai D1, Patino R2, Longoria JA2, Sato R2,
Brenner M1,2 1Univeristy of California, Irvine, Irvine, CA and 2University
of California, Irvine, Irvine, CA.
Purpose of Study: Cyanide (CN) poisoning induces lethal histotoxic anoxia
and stops aerobic cell metabolism by binding to active sites on and disabling
the function of cytochrome c oxidase (CcO). When bound, the electron transport chain is arrested with the near infrared optically active copper core in
reduced form. We developed a broadband diffuse optical spectroscopy (DOS)
prototype system to measure bulk tissue chromophore concentrations and
demonstrated that DOS can quantitatively assess tissue in vivo onset of CN
toxicity and hemorrhage. In this study, we investigated feasibility of DOS
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
detection of CcO redox states by subjecting animals to respiratory challenges
during CN poisoning in a sublethal rabbit cyanide model and swine hemorrhage model where the relationship between hemoglobin and CcO should be
divergent, thus enabling assessment of potential optical CcO signal cross-talk
effects.
Methods Used: New Zealand White rabbits were used in this study. 10mg
of sodium cyanide in 60cc normal saline was infused through the femoral
vein at a rate of 1cc/min. Respiratory challenges, changes in inspired O2 levels
from 100% to 21% and back to 100%, were applied before, during and after
CN infusion, and changes in oxy- and deoxyhemoglobin concentrations and
CcO redox states were monitored continuously with the DOS probe placed
on the right inner thigh muscle. Four hemorrhage studies were conducted
on swine (È25kg). Blood volume (30cc/kg) was removed over 20 minutes
through the right femoral artery. DOS measurements were taken with the
probe placed on the left inner thigh muscle.
Summary of Results: During the hemorrhage, CcO decreased concurrently
with the decrease in oxyhemoglobin indicating that CcO redox state became
reduced due to hypoxia. During CN infusion, CcO redox state also became
reduced while oxyhemoglobin concentration increased due to histotoxic anoxia
and resulting blockage in the electron transport chain.
Conclusions: Spectral similarities between hemoglobin CcO redox states
make non- invasive spectroscopic distinction difficult. By a combination of
physiological perturbations, DOS demonstrates detection of CcO redox state
changes that are decoupled from hemoglobin concentration changes.
154
ONTOGENY AND HIGH ALTITUDE INFLUENCE CA2+
SPARKS IN SHEEP PULMONARY ARTERIAL MYOCYTES
Hadley SR1, Blood Q1, Le P1, Longo LD1, Buchholz J3, Papamatheakis
DG2, Smith GD4, Wilson SM1,2 1Loma Linda University, Loma Linda, CA;
2
Loma Linda University, Loma Linda, CA; 3Loma Linda University, Loma
Linda, CA and 4College of William and Mary, Williamsburg, VA.
Purpose of Study: Ryanodine receptors (RYR) and associated Ca2+ spark
events are important to pulmonary arterial (PA) tone with potential differences in their role in fetuses as compared to adults. RyRs are implicated in
hypoxic pulmonary vasoconstriction (HPV) and ET-1 dependent contractility.
Both HPV and ET-1 signaling are implicated in the development of pulmonary hypertension. Evidence also indicates HPV and PA tone are dysregulated by chronic hypoxia (CH) in fetuses and adults, and yet their combined
influence on Ca2+ sparks and the relationship to PA tone is not known.
We therefore tested the hypotheses that Ca2+ sparks are restricted before
birth and that CH augments their function.
Methods Used: These hypotheses were tested on PA from near term fetal
sheep or adult ewes at low altitude or following 100+ days at 3801 m (CH).
Ca2+ sparks were measured in myocytes of fluo-4 loaded PA strips.
Summary of Results: Ca2+ spark activity was far greater in adults relative
to fetuses while CH inhibited spark activity in fetuses. Ca2+ sparks in pulmonary myocytes from adults had greater amplitudes as compared to fetuses
and the sparks were wider and longer. Chronic hypoxia in comparison reduced
spark amplitude but increased spark width and duration and slowed the time
to peak amplitude. Selective RyR1 inhibition with 10 KM dantrolene reduced
spark activity in normoxic fetuses and adults but not CH adults, while 10 KM
ryanodine restricted spark activity in all three of these groups. Ca2+ spark activity was unaffected by dantrolene or ryanodine in myocytes from CH fetuses.
Overall, RyR function appears developmentally regulated and altered by CH.
Conclusions: The influence of chronic hypoxia and maturation on the role
of RYR activity during HPV remains unresolved, but this data suggests
that RYR function may be blunted in fetus and absent in hypoxic fetus. The
differential effect of CH on RYR activity in fetus and adult provides evidence
that RYRs could be therapeutically relevant in the treatment of pulmonary
hypertension.
155
EFFECTS OF ISCHEMIC CONDITIONING AND ALTITUDE
ON BRACHIAL ARTERY FLOW MEDIATED DILATION
Giri PC1, Matus B1, Rodgers D1, Saleh M1, Lin X3, Foster G1,3, Anholm JD1,2
1
Loma Linda University, Loma Linda, CA; 2VA Loma Linda Healthcare, Loma
Linda, CA and 3VA Loma Linda Healthcare, Loma Linda, CA.
Purpose of Study: Flow mediated dilation (FMD) of the brachial artery is
used to assess endothelial function. Attenuated FMD correlates with adverse
131
Western Regional Meeting Abstracts
Journal of Investigative Medicine
cardiovascular events. Many factors affect flow-mediated vascular reactivity
including plasma nitric oxide (NO) levels. Ischemic conditioning of the extremity (ICE) has cardio-protective effects and one of the mechanisms may
be via NO. NO is also important in the physiologic responses to high altitude
(HA) exposure but the effect of HA on FMD is unknown. The purpose of this
study was to evaluate the effects of ICE and HA on FMD.
Methods Used: FMD was measured in 13 healthy runners at baseline, after
daily ICE or sham-ICE at sea level (SL) and 60 minutes after completing a
12.8km competitive run to the summit of White Mountain, CA at 4342m.
Each subject completed the course twice (with ICE and with sham-ICE)
separated by 6 weeks in a randomized cross-over design. ICE was administered using blood pressure (BP) cuffs around the thigh inflated to 200mmHg
for 5 minutes, deflated for 5 minutes and repeated for 4 cycles. The cuffs
were inflated to 40mmHg for sham-ICE. This was done daily for 5 days at
SL before ascent. Duplex ultrasonography with a linear array transducer was
used to measure brachial artery diameter before and after 5 min of brachial
artery occlusion. The effects of HA and ICE on FMD were evaluated using
repeated-measures ANOVA.
Summary of Results: Complete data were obtained in 12 subjects.
At SL, FMD was 15.5% T 7.0% (mean T SD) without ICE and 9.8% T 6.1%
with ICE. At HA, FMD was 11.5% T 9.5% without ICE and 6.0% T 5.1%
with ICE. Compared to SL without ICE, FMD was attenuated by 36.5% with
ICE at SL, 60.7% with ICE at HA and 25.5% without ICE at HA. Both ICE
and HA blunted the FMD response (pG0.05 for ICE and PG0.01 for HA by
ANOVA, ICE*HA interaction p=NS). Compared to SL without ICE, preinflation diameters were larger by 10.1% with ICE at HA and by 3.6% without
ICE at HA.
Conclusions: FMD is significantly attenuated by both ischemic conditioning of extremity and high altitude. This finding is predominantly due to larger
pre-dilation artery diameters likely induced by vasodilators released by ICE
and altitude exposure.
156
COMPARISON OF SVV TO SCVO2 IN PATIENTS
UNDERGOING MAJOR SURGICAL PROCEDURES
King NC, Blair BM, Collier C, Applegate RL, Sanghvi C, Allard M
Loma Linda University Medical Center, Loma Linda, CA.
Purpose of Study: Major surgical procedures are accompanied by a risk
of intraoperative blood loss and fluid shifting. Prior studies have demonstrated that intraoperative goal directed fluid therapy based on keeping stroke
volume variation (SVV) below a critical threshold might result in increased
cardiac output, stroke volume, and blood pressure. It has also been shown
that increasing low central venous oxygen saturation (ScVO2) with fluid
therapy in sepsis patients is associated with improved patient outcome. This
study was designed to investigate the relationship between SVV and ScVO2
in the intraoperative, non-emergent, surgical setting.
Methods Used: This is a Departmentally sponsored, IRB approved prospective non-randomized sequential study in adult patients undergoing major
surgical procedures with a pre-operative estimated blood loss of Q15% total
blood volume. SVV was measured using an arterial pressure cardiac output
device (Vigileo, Edward Lifesciences, Irvine CA). ScVO2 was continuously
monitored using an oximetric central venous catheter (PreSep, Edward Lifesciences, Irvine CA). Data from these devices was collected through an electronic
interface to the electronic anesthesia record. Intraoperative fluid management was guided to maintain SVV G12%. Transfusion of blood products
was at the discretion of the physicians caring for the patient. Data was analyzed for correlation between SVV and ScVO2 using JMP V8.0.2, with
pG0.05 as statistically significant.
Summary of Results: Data was collected from 34 subjects. A total of
16187 time matched SVV to ScVO2 measurements were used for comparison. There was no correlation between SVV and ScVO2 paired samples
(Pearson_s r = j0.148; r2 of 0.022).
Conclusions: It has been reported that SVV is useful for guiding intraoperative fluid therapy, while ScVO2 has been used to guide fluid therapy in
sepsis patients. This study shows no correlation between SVV and ScVO2
during the intraoperative phase of patient care. Although SVV and ScVO2
can be used independently as guides for fluid therapy in different settings,
intraoperative ScVO2 appears to not be interchangeable with SVV for
guiding intraoperative fluid therapy.
132
&
Volume 59, Number 1, January 2011
References:
Crit Care Cli. 2010;26:323
N Engl J Med. 2001;345:1368
J Intensive Care Med. 2009;24:352.
157
VITAMIN D RECEPTOR DELETION DISRUPTS NORMAL
LUNG DEVELOPMENT AND LEADS TO AN ASTHMA
PHENOTYPE
Li Y1, Kovacs C2, Tou E1, Satyadev N1, Torday J1, Rehan V1 1Los Angeles
Biomedical Institute, Torrance, CA and 2 Memorial University of
Newfoundland, St. John’s, NF, Canada.
Purpose of Study: There are strong epidemiologic data suggesting a link
between vitamin D deficiency during pregnancy and childhood asthma.
These data are complemented by experimental animal data showing that
vitamin D is one of the local alveolar paracrine factors that spatiotemporally modulates perinatal pulmonary maturation. We have recently shown
that vitamin D augments perinatal lung maturation such that its deficiency
would perturb normal lung structural and functional development in a way
that is consistent with the asthma phenotype. However, the mechanistic link
between vitamin D deficiency during pregnancy and childhood asthma is not
fully established. In this study, we aimed to determine the effect of vitamin D
receptor deletion on key markers of lung maturation.
Methods Used: Control (wild type) and vitamin D receptor null mutant
mice (C57BL6 background) were examined at 2 and 7 weeks postnatally.
At sacrifice, lungs were perfused with phosphate buffered saline and collected for morphometry, RNA and protein analyses. Western blot and immunohistochemistry for key markers of lung differentiation were performed.
Furthermore, using the Affymetrix Mouse Genome 430 2.0 array, the transcriptional profile of the lungs from the two groups at 7 weeks of age was
determined.
Summary of Results: Both by Western analysis and by immunohistochemistry, at 2 and 7 weeks of age there was down-regulation of the intermediates for PTHrP/PPARF signaling, a key signaling pathway for lung
maturation, and up-regulation of Wnt signaling intermediates, a key pathway
in asthma pathogenesis. Self-Organizing Map cluster analysis corroborated
these data by showing up-regulation of the Wnt signaling pathway, and downregulation of the PTHrP/PPARF signaling pathway in vitamin D receptor null
mice compared to WT mice.
Conclusions: These data suggest that altered vitamin D signaling leads
to down-regulation of homeostatic PPARF signaling, and up-regulation of
Wnt signaling, predisposing to a myogenic pulmonary phenotype that is
consistent with the propensity to asthma in vitamin D deficiency.
158
SIMVASTATIN INHIBITS EXPRESSION OF EOTAXINS
IN MOUSE AND HUMAN AIRWAY EPITHELIUM:
IMPLICATIONS FOR THE TREATMENT OF
ALLERGIC ASTHMA
Zeki AA1,2, Thai P1,2, Wu R2 1U.C. Davis Medical Center, Sacramento, CA
and 2Center for Comparative Respiratory Biology & Medicine (CCRBM),
Davis, CA.
Purpose of Study: We have previously shown that systemic administration
of simvastatin in a mouse model of allergic asthma inhibits eosinophilic
inflammation and improves lung function. We wanted to evaluate the potential
contribution of the airway epithelium to this observation. We hypothesized
that simvastatin attenuates the expression of Th2 cytokines/chemokines important in human eosinophilic asthma.
Methods Used: Human bronchial epithelial cells (HBE1) were grown under
air-liquid interface (ALI) conditions until confluent. Mouse tracheal epithelial
cells were harvested from naBve BALB/c mice and grown to confluence in ALI
for 4 weeks. HBE1 cells were pre-treated with simvastatin (Sim, 20 KM) for
3 days, then stimulated with IL-13 (20 ng/mL) for 6 hours on the last day. Mouse
cells were initially pre-treated with Sim 10 KM for 24 hours, then stimulated
with IL-13 (20 ng/mL) and co-incubated with Sim for 48 hours. Expression
of chemokines eotaxin-1, eotaxin-3, MCP-1, MCP-2, MCP-3, and CCR3 was
analyzed by RT-PCR. Gene expression was assessed relative to housekeeping
genes Hsp90ab1 (for mouse cells) and A-Actin (for HBE1 cells).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: In HBE1 cells, Sim treatment reduced IL-13induced eotaxin-3 expression by 58% (pG0.00089), and eotaxin-2 expression
by 30.6% (p=0.111). In mouse cells, Sim treatment reduced IL-13-induced
eotaxin-1 expression by 92.6% (pG0.05), MCP-1 expression by 84% (pG0.005),
MCP-2 expression by 53.8% (p=NS), and MCP-3 expression by 87.2%
(pG0.05). Simvastatin (10 KM) also attenuated the mRNA expression of CCR3
by 62.03%, the receptor for chemokines important in eosinophil recruitment
(i.e. RANTES, MCPs, and eotaxins).
Conclusions: These data suggest that simvastatin attenuates Th2 cytokines/
chemokines important in eosinophilic inflammation pertinent to allergic
asthma. At least some of the simvastatin effect may be occurring at the level
of the airway epithelium. Thus, statins may be a novel therapy for allergic
asthma that warrants additional study in humans.
Supported by the following grants: NIH (T32) HL07013, NCRR UL1
RR024146 (K30), HL-076415 (K08), ATS Fellows Career Development
Award, and CTSC K12 Award (KL2 RR 024144).
159
DEVELOPMENT OF A COBINAMIDE-BASED CYANIDE
SENSOR TO RAPIDLY DETECT CYANIDE TOXICITY
Longoria JA1, Vo A2, Blackledge W3, Yoshii I1, Le T1, Liou R1, Mahon S2,
Boss G3, Brenner M1,2 1University of California Irvine Medical Center,
Orange, CA; 2University of California Irvine, Irvine, CA and 3University
of California San Diego, San Diego, CA.
Purpose of Study: Cyanide is an extremely fast acting, potent and lethal
agent and exposure may result from a number of different events including
smoke inhalation, acts of terrorism and industrial accidents. Death can occur
within minutes of exposure. Therefore, rapid diagnosis of cyanide toxicity
is of great importance. Current methods to detect and quantify cyanide in
blood, such as mass spectrometry, high-performance liquid chromatography,
and spectrophotometry are slow (hours to perform), as they require offgassing and trapping of cyanide in HCN gaseous phase. Here we present a
simple, whole blood assay employing cobinamide and spectrophotometry
without the use of toxic reagents or off-gassing of cyanide to rapidly detect
cyanide in blood samples.
Methods Used: We used cobinamide, a precursor to cobalamin biosynthesis with an overall cyanide binding affinity of 1022 Mj2, to bind with cyanide
in whole blood to extract cyanide from erythrocytes. After cyanide was added
to whole blood to simulate cyanide exposure, cobinamide was added to the
specimen and mixed for 3Y5 minutes. Plasma was then separated and absorbance was measured with a spectrophotometer and assessed for characteristic
spectral changes of resultant dicyanocobinamide.
Summary of Results: When cobinamide binds to cyanide and forms
dicyanocobinamide, a peak at 580 nm wavelength in absorbance predictably
shifts based on relative cobinamide/dicyanocobinamide concentrations in
the specimen. Results were reproducible in a rapid whole-blood assay requiring minimal equipment and non-toxic reagents.
Conclusions: This study demonstrates feasibility of concept for development of a rapid cobinamide-based cyanide assay for detecting cyanide levels
and toxicity in whole blood. With completion of standard assay curves, accurate, rapid cyanide assays using cobinamide and simple spectrophotometry
should be possible using this approach. In the future development of Bdipstick[
based rapid field diagnostic testing kits may be possible using these principles.
160
TEMPORARY ARM OCCLUSION MAY REVEAL LEVEL
OF CYANIDE TOXICITY
Kim JG1, Lee J1, Mahon S1, Mukai D1, Yoon D1, Brenner M1,2 1University
of California Irvine, Irvine, CA and 2University of California Irvine Medical
Center, Orange, CA.
Purpose of Study: Previously, we found that amplitudes of non-invasively
detected oxy (OHb)/deoxyhemoglobin (RHb) concentration changes during
respiratory challenge from 100% to 21% oxygen decrease as acute cyanide
(CN) toxicity increases in an animal model. Because respiratory challenge
cannot be used in CN exposed patients since oxygen supplementation is
critical, we tested an alternative physiologic stress for estimating CN level
using temporal occlusion on the upper arm.
Methods Used: New Zealand white rabbits were administered saline (n=5),
10 mg of NaCN (n=6) or 20mg of NaCN (n=6) in 60ml saline via the femoral vein at 1ml/min. After CN infusion, additional 90min of recovery was
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
monitored. Upper arm occlusions using a finger cuff (130mmHg) were
applied for 4 mins, then released before, during, and post cyanide infusion.
Changes in OHb and RHb concentration throughout the experiment were
measured from lower arm muscles using near infrared spectroscopy (NIRS).
The decrease/increase rate of OHb and RHb concentration during occlusion
was determined by calculating the half-time constant,T1/2, using a Hill equation [OHb or RHb = A(tn/(T1/2+ tn))] where A is amplitude of OHb and RHb
changes and t is time.
Summary of Results: Tourniquet application caused a drop of OHb and
increase of RHb, which were reversed as the tourniquet pressure was released. Amplitudes of both OHb and RHb changes lessened during cyanide
infusion and returned to the baseline level at the end of recovery while
control animals showed little changes. 20mg CN infused animals showed a
much greater drop in the amplitudes of both OHb and RHb changes than
10mg CN infused animals (50% vs 25% drop compared to the baseline).
20mg CN infused animals also showed that OHb decrease rate during the
upper arm occlusion was 3.5 times slower compared to the baseline values
during cyanide infusion, compared to 1.5 times from 10mg CN infused
animals.
Conclusions: These results support our hypothesis and show that NIRS
combined with temporal occlusion may be a potential non-invasive tool for
estimating extent of cyanide toxicity in vivo.
161
LETHAL LEVEL CYANIDE POISONING REVERSAL
BY INTRAMUSCULAR INJECTION OF COBINAMIDE
Yoon D1, Lee J1, Kim JG1, Mukai D1, Mahon S1, Boss G3, Blackledge W3,
Brenner M2 1Beckman Laser Institute, Irvine, CA; 2University of California
San Diego, La Jolla, CA and 3University of California Irvine, Irvine, CA.
Purpose of Study: Cyanide poisoning is a major worldwide threat. Military and civilian mass exposures to cyanide may occur through acts of
terrorism and war. Current effective treatment methods, which require intravenous delivery, are ineffective in mass casualty situations. We are investigating
intramuscular administration of Cobinamide, a novel water-soluble molecule
that can bind 2 molecules of cyanide per molecule with higher affinity than
currently approved hydroxocobalamin, for the treatment of mass casualty cyanide poisoning. In this study, we have evaluated the efficacy of IM cobinamide
formulations in a rabbit model of cyanide poisoning.
Methods Used: Lethal level cyanide toxicity was generated in anesthetized and ventilated New Zealand white rabbits by infusion of 20mg cyanide solution intravenously over 60min. At the time of treatment, animals
were injected intramuscularly with specific formulations of oxidized and
reduced cobinamide, and compared to the control saline injected group.
Quantitative analysis was done using continuous wave near infrared spectroscopy (CWNIRS) and diffuse optical spectroscopy (DOS) for CNS region
and tissue oxy and deoxyhemoglobin concentrations. Arterial blood was
collected to measure the level of cyanide, blood gasses and cobinamide.
Summary of Results: IM injection of specific reduced cobinamide formulations has demonstrated the ability to reverse the effects of lethal CN
poisoning, including fast recovery of oxy and deoxyhemoglobin concentrations in cyanide exposed animal tissue. Blood plasma samples also indicated
significant amounts of cobinamide absorption into the blood.
Conclusions: The result indicates that cobinamide is a rapid acting agent
and appears to be effective in reversing the physiologic effects of cyanide
poisoning when given intravenously to lethally CN poisoned subjects. Our
additional study suggested that sulfur containing reducing agents may facilitate
transportation of cobinamide into the blood more effectively. Future investigations on other additives that will allow rapid cobinamide transport are needed.
162
POSTNATAL MATURATION DECREASES THE ROLE
OF RHO-KINASE IN ELECTROMECHANICAL COUPLING
OF SHEEP PULMONARY ARTERIES
Papamatheakis D1, Vemulakonda S1, Patel J2, Blood Q2, Merritt T2, Longo
LD2, Wilson SM2,1 1Loma Linda University Medical Center, Loma Linda,
CA and 2Loma Linda University, Loma Linda, CA.
Purpose of Study: L-type Ca2+ channels (CaL) as well as Rho-kinase are
important in electrically induced contraction of pulmonary arteries, thus being
therapeutically relevant in pulmonary hypertension. Evidence indicates that the
role of CaL in pulmonary artery contraction is potentially reduced by chronic
133
Western Regional Meeting Abstracts
Journal of Investigative Medicine
hypoxia (a risk factor for pulmonary hypertension), but increased after birth.
In comparison, the role of Rho-kinase may increase following chronic hypoxia
and decrease after birth. We tested the hypotheses that CaL and Rho-kinase
coupling is blunted by chronic hypoxia and accentuated following birth.
Methods Used: We examined isometric contraction using wire myography
in pulmonary arterial rings isolated from near term sheep fetuses (È140 days
gestation) and adult ewes maintained under normoxic (300 m altitude) or
chronic hypoxic (3801 m) conditions for 100+ days. Pulmonary arterial rings
were depolarized with cumulative doses from 5 to 125 mM of K+, or stimulated repeatedly with 125 mM of K+.
Summary of Results: One-hundred and twenty-five mM of K+ elicited
the greatest force in pulmonary arteries from adults and this was not influenced by chronic hypoxia. The estimated EC50 for K+-induced contraction
was similar in pulmonary arteries from all four animal groups. CaL inhibition
with 10 KM verapamil, diltiazem or nifedipine reduced, but did not ablate,
K+-dependent contraction. Rho-kinase inhibition with 10 KM Y27632 reduced K+ contraction to a greater extent in pulmonary arteries from fetuses
as ompared to adults regardless of their gestational altitude. The combination of 10 KM nifedipine and 10 KM Y27632 dramatically reduced K+ contraction in both fetuses and adults.
Conclusions: Overall, electromechanical coupling appears augmented by
maturation and unaffected by chronic hypoxia. Moreover Rho-kinase related pathways appear to contribute to this coupling process in a maturational
dependent manner. This adds to a growing body of evidence regarding the
importance of Rho-kinase in the treatment of pulmonary hypertension in
newborns and adults.
163
COUGH AND CHEST BULGE: A RARE CASE
OF SPONTANEOUS LUNG HERNIATION
Giri PC, Anholm JD Loma Linda VA Healtcare System, Loma Linda, CA.
Case Report: Spontaneous lung herniation is a rare but often underdiagnosed entity.
History: 69 year old male with COPD, Gold Stage III presented with cough
and shortness of breath for three weeks. He had an acute onset of right sided
chest pain and swelling one week prior to admission after he felt Bsomething
give away[ in his back.
Physical Exam: Chest wall: 10 x 10 cm bulge below right scapula, prominent with cough. Ecchymoses seen in lower half of chest wall.
Imaging: CT scans showed herniated lung through the right 8th intercostal
space.
Discussion: Less than 300 cases of lung herniation have been reported of
which around a 100 are spontaneous thoracic (intercostal) hernias. Most
result from an acute increase in intrathoracic pressure due to coughing,
sneezing, blowing on a musical instrument or heavy lifting classically in
&
Volume 59, Number 1, January 2011
male smokers with chronic pulmonary disease. The diagnosis is usually
confirmed by means of chest radiography, fluoroscopy or computer tomography. It is sometimes missed on routine chest radiographs.
Management: Complications (strangulation, incarceration) are rare. Conservative management is recommended. Spontaneous resolution seldom occurs.
Compressive pads or corsets may be effective for pain. Surgery is curative and
is indicated when there is pain, recurrent infection, hemoptysis, interference
with daily activities, if complications arise or if the hernias are very large.
Outcome: This patient was seen by thoracic surgery and managed conservatively. He denied symptoms during follow up in clinic.
Surgery I
Concurrent Session
8:30 AM
Friday, January 28, 2011
164
EFFECT OF KIDNEY REST ELEVATION ON
POSTOPERATIVE CREATININE KINASE LEVELS IN
HAND-ASSISTED LAPAROSCOPIC DONOR NEPHRECTOMY
Peters A, Agarwal G, Richards G, Tenggardjaja C, Bowman R, Baldwin D
Loma Linda University, Loma Linda, CA.
Purpose of Study: Rhabdomyolysis (RM) during minimally invasive surgery has been reported in the literature and is a known cause of acute renal
failure. Known risk factors in the perioperative period include positioning,
operative time, and patient size. Our objective was to evaluate whether kidney
rest elevation has an effect on postoperative creatinine kinase (CK) levels and
risk for RM.
Methods Used: Serum CK was prospectively measured in all kidney
donors at a single academic institution between February of 2003 and April
of 2010. Thirty-one patients who underwent donor nephrectomy without
kidney rest elevation were compared to forty patients who underwent donor
nephrectomy with kidney rest elevation. Statistics were performed using either
an independent sample t-test or a Mann-Whitney test, followed by multiple
linear regression to determine independent variables with p G0.05 considered
significant.
Summary of Results: Patient demographics and preoperative characteristics were not significantly different between the two groups. Laparoscopic
donor nephrectomy with the kidney rest elevated was not a significant predictor of increased postoperative CK values compared to patients without
kidney rest elevation (median CK 450 vs 458 IU/L; p = .871). Kidney rest
elevation did not result in increased postoperative creatinine compared to
patients without kidney rest elevation during laparoscopic donor nephrectomy
(median creatinine 1.18 vs 1.17 mg/dL; p = .809). After adjusting for length of
surgery and gender, the kidney rest was not a significant predictor of increased
postoperative CK values (p = .834).
Conclusions: Kidney rest elevation does not affect the risk for increased
postoperative CK and RM. Other factors, including surgeon preference and
comfort with the surgical position, should determine whether or not the
kidney rest is elevated during laparoscopic donor nephrectomy.
165
ADIPOSE DERIVED-STEM CELL ADHERENCE,
PROLIFERATION, AND MIGRATION ON
ALLODERM\ MATRIX
Slack GC1,2, Hagvall S1,2, Zuk P1,2, Rahgozar P2, Yaghoubian A2, Kruger E2,
Ehsani N2, Tabit C2, Bradley J1,2 1UCLA David Geffen School of Medicine,
CA, CA and 2UCLA, Los Angeles, CA.
Purpose of Study: The use of acellular dermal matrices, like AlloDerm\
Regenerative Tissue Matrix, is currently used in breast reconstruction after
ablative mastectomy surgery. Adipose derived stem cells (ASCs) are multipotent cells that may differentiate into fat, bone, muscle or nerve cells. The
aim of our study was to determine if AlloDerm\ may act as a suitable
scaffold for ASCs to assist faster integration of local tissue and graft and
promote revascularization, of importance prior to radiation treatment for
breast cancer.
Methods Used: ASCs, obtained from liposuction patients, were seeded
onto the dermal face of the AlloDerm\ scaffold by two methods: 1) pipet
placement, 2) centrifugation (1000 rpm for 5 minutes). Constructs were
134
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
harvested after 2, 5, 7, 14, 21, 28, and 42 days in tissue culture. H&E, DAPI,
and SEM imaging studies for cell attachment, proliferation and migration
at various matrix depths and real time reverse transcriptase polymerase chain
reaction (RT-PCR) for adipogenic and osteogenic differentiation were performed at each time point.
Summary of Results: ASC adherence and viability was confirmed by SEM
imaging with cellular microvilli and interdigitation within the AlloDerm\
scaffold for both seeding methods. Proliferation increased from 16 to 55 mean
per high-powered field in the first week then stabilized. Migration increased
from 13Y18% to 35Y40% penetration into the matrix depth in the first week;
by week 6, some ASC groups were limited to the surface though others
had 100% penetration. Early studies show cultured ASCs retain adipogenic
potential within AlloDerm\ as seen by expression of adipogenic markers by
RT-PCR.
Conclusions: Our findings show that ASCs seeded, by drop or centrifuge
method, onto AlloDerm\ successfully attach and proliferate over a 6-week
period. In vivo studies are being undertaken.
Western Regional Meeting Abstracts
Summary of Results: The iliohypogastric (IHG) nerve was found to be
2.9mm wide, 27.1mm from the twelfth rib (T12), and 46.2- from the lateral
border of the psoas muscle. The ilioinguinal (ILI) nerve was 2.1mm wide,
43.3mm from T12, and 44.6- from the psoas muscle. The genitofemoral
(GEN) nerve, located 93.7mm from T12, was found overlying the psoas
with a width of 2.3mm. The lateral femoral cutaneous (LFC) nerve was
2.2mm wide, 188mm from T12, and 38.1- from the psoas. The 7.7mm
wide femoral (FEM) nerve was located 150.2mm from T12 and 22.9- from
the psoas.
Conclusions: In this study, we observe considerable anatomic variability
associated with each lumbar plexus branch. The results of the study can improve outcomes in surgical intervention of post-herniorrhaphy inguinodynia.
Measurement of Retroperitoneal Lumbar Plexus Nerves
166
COMBINATORY EFFECT OF BMP-2 AND APATITE COATED
SCAFFOLDS ON THE OSTEOGENIC DIFFERENTIATION
OF HUMAN MESENCHYMAL STEM CELLS
Davis HE, Case E, Miller S, Leach JK University of California, Davis,
Davis, CA.
Purpose of Study: It is widely accepted that osteoconductive materials
promote osteogenesis in vivo. However, the impact of 3D osteoconductive
substrates coupled with soluble signals on progenitor cell differentiation is
not well known. In this study, we investigated the influence of carbonate
apatite on the osteogenic differentiation of human mesenchymal stem cells
(hMSCs) seeded in biodegradable poly(lactide-co-glycolide) (PLG) scaffolds
in conjunction with the osteoinductive growth factor BMP-2.
Methods Used: Apatite coated PLG scaffolds were formed by incubating
hydrolyzed microparticles in modified simulated body fluid (mSBF) prior
to being combined with NaCl and compressed at 1500 psi for 1 min. Samples
were then exposed to high pressure CO2 (800 psi) for 16 h. NaCl was
removed by leaching the matrices in distilled H2O for 24 h. We cultured
hMSCs on the scaffolds in osteogenic media supplemented with 25, 100
or 200 ng/ml of BMP-2. The in vitro osteogenic differentiation of hMSCs
was measured by quantifying ALP activity, DNA content, and cell secreted
calcium after 3, 7, 14, 21 and 28 d in osteogenic conditions. qPCR was
performed for Runx2, SP7, Col1a1, lbsp, and Sparc at 7 and 21 d.
Summary of Results: No significant differences between nonmineralized
and mineralized scaffolds were detected in ALP activity at 0 and 200 ng/mL
concentrations, but mineralized scaffolds had increased levels after 7 d at
25 and 100 ng/mL. Mineralized scaffolds had increased levels of cell-secreted
calcium at all time points. Increasing BMP-2 concentration resulted in a rise
in both ALP and calcium levels. There was no difference between Runx2
and SP7 levels in mineralized and nonmineralized substrates, but an increase
in SP7 was observed with the addition of BMP-2. At 21 d mineralized substrates had increased levels of lbsp and decreased levels of Sparc compared
to nonmineralized substrates.
Conclusions: The present data indicate that a combination of apatite and
BMP-2 do not simply enhance the osteogenic response of hMSCs, but act
through different and possibly opposing pathways. Thus multiple signaling
strategies may be necessary to achieve optimal bone regeneration.
167
RETROPERITONEAL ANATOMY: IMPLICATIONS
FOR INGUINODYNIA
Mannan AS1, Walker A1, Stark E2, Towfigh S3 1David Geffen School of
Medicine at UCLA, Los Angeles, CA; 2David Geffen School of Medicine at
UCLA, Los Angeles, CA and 3Cedars-Sinai Medical Center, Los Angeles, CA.
Purpose of Study: We aim to quantify anatomy of lumbar plexus nerves
to assist in safe performance of laparoscopic triple neurectomy for treatment
of post-herniorrhaphy inguinodynia.
Methods Used: Measurements were obtained bilaterally from 9 female and
6 male cadavers provided by the UCLA Donated Body Program. Distances
from the twelfth rib were measured to where each nerve emerges from the
psoas muscle.
* 2011 The American Federation for Medical Research
Figure 1. Schematic Representation of Retroperitoneal Lumbar
Plexus Nerves
168
TELEMEDICINE PRE-ANESTHESIA CONSULTATION
PROVIDES ACCURATE AIRWAY EXAM INFORMATION
Wolford B, Gildea B, Applegate RL, Stier G, Patchin R, Nyirady J LLU,
Loma Linda, CA.
Purpose of Study: Telemedicine has potential to reduce cost and patient
inconvenience in pre-anesthesia consultation (PAC)3. A pilot report2 described
overall patient and provider satisfaction with pre-anesthesia telemedicine consultation (PATC), but did not compare airway exam findings from PATC with
day of surgery findings. This IRB approved randomized study evaluates whether
risk factors and potential complications of perioperative anesthesia care can be
accurately identified by PATC.
Methods Used: We compared PAC and satisfaction surveys from three
groups (phone, in-person PAC, and PATC) to day of surgery evaluations.
Utilizing criteria-based screening, we assigned adult otolaryngology patients
with no history of anesthesia/airway/major medical complications to the phone
interview group. Others were randomized to PAC, or PATC groups. PATC
telemedicine consultation was done at the otolaryngology clinic, with an anesthesia provider at our pre-anesthesia clinic. PATC employed two-way videoconference camera and monitor, digital electronic stethoscope, and handheld
airway exam camera, operated by an otolaryngology clinic nurse who had been
provided brief training on the airway evaluation. All consultations included
preoperative history and surveys noting anticipated complications or surgical
135
Western Regional Meeting Abstracts
Journal of Investigative Medicine
delays. Providers also conducted ASA 11-pt airway, heart, and lung exams
for the PAC and PATC groups. Laryngoscopic view was graded, with Grade 3
or 4 considered difficult view (DV). Difficult airway management (DA) was
defined as 91 attempt by an experienced provider and/or the need for 91 type
of laryngoscope.
Summary of Results: 174 participated (38 phone, 64 PAC, 72 PATC).
Prediction of DV by PAC and PATC correlated well with the Staff Anesthesiologist’s evaluation on day of surgery. 19 patients (10 PAC; 19 PATC)
had DV: PAC and PATC were equally likely to predict this. DA was present
in 6: the Anesthesiologist’s evaluation was incrementally more predictive
of this (67% vs 50%). No unexpected findings were noted on heart/lung
exams on day of surgery, and patients/providers reported high satisfaction
with PATC.
Conclusions: PATC is technically feasible and equivalent to current inperson evaluation, including the ASA 11-point airway exam, with additional
potential to minimize patient inconvenience and costs of missed/cancelled
appointments.
169
FAILURE STRENGTH OF TENDON REPAIR WITH
VARYING STITCH COUNT IN THE PULVERTAFT
WEAVE AND SIDE-TO-SIDE TECHNIQUES
Adair CR2, Williams P12, Inceoglu S1, Wongworawat MD1 1Loma Linda
University, Loma Linda, CA and 2Loma Linda University School of
Medicine, Loma Linda, CA.
Purpose of Study: The Pulvertaft weave (PTW) is a common repair
technique used during tendon transfers. This study was designed to compare the maximum load-to-failure of the PTW and side-to-side (STS) repair
methods and to analyze the mechanical interaction between the number of
weaves and stitches.
Methods Used: Six groups of repairs were performed using a porcine
tendon model. Three PTW groups, PTW3, PTW4, and PTW5, had one, two,
and three weaves, respectively (N=5). The others, utilizing STS technique
(STS3, N=5; STS4, N=5; STS5, N=7), had no weaves, but equal amounts
of stitches as in each PTW group. For PTW repairs, incisions were made
at the tendon free end and the transfer tendon weaved through the hosting
tendon. A stitch was placed at the free ends and at each weave point. One
weave yielded three stitches with an additional stitch per additional weave.
Overlapping the tendons and placing three to five equidistant stitches completed the STS repairs. Cross-stitches with four stitch points were used for
all repairs. The specimens were tested to failure in tension using an Instron
machine.
Summary of Results: All failures occurred within the repair region.
The mode of failure for all groups was suture breakage. There were no
significant differences among the three STS groups in maximum load-tofailure (STS3, 239T16.4 N; STS4, 262T19.1 N; STS5, 253T91.5 N). PTW4
(402T45.8 N) was not significantly different than PTW3 (309T21.4 N, p=0.08)
and PTW5 (501T50.8 N, p=0.06). However, PTW5 was greater than PTW3
(pG0.05). PTW4 and PTW5 were greater than all STS groups (pG0.05). A linear
regression of the number of weaves versus load for all groups yielded a slope
of 80.84 (R2=0.78, pG0.001). A linear regression of the suture count versus
load-to-failure for STS groups yielded a slope of 1.6 (R2=0.01, p=0.72) and
for the PTW groups yielded a slope of 23.9 (R2=0.82, pG0.001).
Conclusions: The PTW construct had better mechanical properties than
the STS. This study suggests that when using a cross-stitch for tendon
repair, the addition of weaves provides a dramatic increase in maximum
load-to-failure capacity of the construct and that an increase in the number
of stitches provides mechanical benefit only when the tendons are weaved.
170
OCULAR IMAGING TECHNIQUES: AN USEFUL TOOL
IN THE DIAGNOSIS AND MANAGEMENT OF PATIENTS
WITH ANTERIOR SEGMENT PATHOLOGY
Dehdashti N1, Ursea R2 1University of Arizona College of Medicine, Tucson,
AZ and 2University of Arizona College of Medicine, Tucson, AZ.
Purpose of Study: Ocular pathology affecting the anterior segment can
present diagnostic problems because it often involves structures that are
not readily accessible through routine examination methods. The goal of the
study was to assess and compare the utility of ultrasound biomicroscopy
(UBM) and anterior segment optical coherence tomography (AS-OCT) in the
136
&
Volume 59, Number 1, January 2011
evaluation and management of patients with ocular pathology involving the
anterior segment of the eye.
Methods Used: Retrospective review of 40 eyes with various conditions
involving the anterior segment, examined with UBM (Sonomed, Inc, Lake
Success, NY) and AS-OCT (Carl Zeiss Meditec Inc., CA) over a time
period of 18 months.
Summary of Results: Abnormalities were found in 32 eyes of patients
with the following etiologic diagnoses: anterior scleritis, intermediate uveitis,
traumatic hyphema, foreign body, iris or ciliary body mass and intraocular
lens. Indications for the UBM were: visualization of anterior chamber structures, visualization of posterior iris or ciliary body, assessment of intraocular
lens position, pre-operative assessment in eyes with hypotony or trauma and
for follow-up of documented abnormalities. Indications for anterior segment
OCT were: assessment of the angle, measurement of corneal thickness, visualization of anterior iris and preoperatory assessment.
Conclusions: The OCT and UBM imaging techniques offer a complete
assessment of the anterior segment of the eye. Both are sensitive imaging
modalities that provide excellent diagnostic clues in patients with anterior
segment involvement. UBM is invaluable in accurately visualizing the posterior iris and ciliary body region. In the majority of cases the high-resolution
imaging techniques confirmed the clinical diagnosis and were useful in patient
follow up and surgery planning.
171
URETEROSCOPY WITHOUT INTRA-OPERATIVE
FLUOROSCOPIC IMAGING AS A SAFE ALTERNATIVE
Baldwin B, Richards G, Agarwal G, Tenggardjaja C, Bowman R, Baldwin DD
Loma Linda University, Loma Linda, CA.
Purpose of Study: During the course of a ureteral stone episode, a patient
is at risk for multiple doses of radiation. Although the doses are not routinely high enough to cause dose-dependent effects such as skin burns, the
dose-independent effects such as cancer, could potentially be severe, warranting the reduction of radiation exposure whenever possible. This retrospective
study assesses the safety and effectiveness of ureteroscopy for the treatment
of kidney and ureteral stones without the use of fluoroscopic guidance in light
of the potential risks to patients and medical personel that are inherent to radiation exposure.
Methods Used: A retrospective review of ten patients who electively received ureteroscopy without intra-operative imaging between January 2009
and June 2010 was performed and the outcomes of the procedures were
analyzed to determine their effectiveness. These ten patients and their outcomes were then compared with 25 patients who received standard low-dose
fluoroscopy-guided ureteroscopy during that same time period.
Summary of Results: The two groups were found to have no significant differences in gender, age, BMI, stone laterality, stone size, stone location, postoperative complication rate, stone free rate, or repeat procedures
(all P 9 0.45). The greatest difference found was that operating time for the
fluorless precedure was 9.7 minutes shorter than for the low-dose procedure
(p = 0.26).
Conclusions: In select patients, a fluoroless ureteroscopy technique can
safely and effectively be employed by a skilled physician for the treatment
of kidney and ureteral stones without exposing the patient to unnecessary
and potentially harmful radiation.
172
RECONSTRUCTIVE OR COSMETIC PLASTIC SURGERY:
FACTORS INFLUENCING THE TYPE OF PRACTICE
ESTABLISHED BY CANADIAN PLASTIC SURGEONS
McInnes C1, Courtemanche D2, Verchere C2, Bush K2, Arneja J2 1University of
British Columbia, Vancouver, BC, Canada and 2British Columbia Children’s
Hospital, Vancouver, BC, Canada.
Purpose of Study: Recently, some in organizational plastic surgery have
voiced concerns that the specialty is facing an identity crisis. Challenged
by factors such as increasing competition in the cosmetic marketplace and
decreasing reimbursement for reconstructive procedures, many American
plastic surgeons have adopted increasingly cosmetic-focused practices. This
study will investigate the currently unknown practice profiles of Canadian
plastic surgeons to determine the reconstructive-cosmetic mix, as well as
factors which influence the type of practice to see if a similar pattern is
occurring in Canada.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Methods Used: An anonymous online survey was distributed to all 352
Purpose of Study: Lumbar spinal stenosis is a disabling medical condi-
Canadian plastic surgeons with email accounts registered with the Canadian
Society of Plastic Surgeons (CSPS) and/or the Canadian Society for Aesthetic Plastic Surgery (CSAPS) which contained questions regarding practice
profiles.
Summary of Results: A 34% response rate was found (120 responses),
of which 76% have reconstructive practices and 24% have cosmetic practices. Variables more common among reconstructive surgeons were female
gender, younger age, pre-residency research experience, advanced degrees
(Master_s/PhD) and higher educational debt, among others.
Conclusions: The field of reconstructive plastic surgery appears to be
thriving in Canada. Compared to the US, a greater proportion of Canadian
plastic surgeons have reconstructive practices, likely due to different funding models and competition. Compared to cosmetic surgeons, a greater
proportion of reconstructive surgeons hold university positions, have more
academic practices, work longer hours, and take more emergency call. When
deciding which type of practice to establish, reconstructive surgeons were
relatively more influenced by academic opportunities, and less influenced by
financial and non-financial metrics.
tion in which narrowing of the spinal canal compresses the spinal cord and
nerves. Entrapment of the cauda equina roots often presents with difficulty
walking, pain in the back and lower extremities, and weakness in the legs,
a condition called neurogenic intermittent claudication (NIC). The disease
continues to be a leading cause of morbidity in the elderly population and
is the leading cause of lumbar spine surgery for adults over the age of 65.
Historically, the standard operative procedure to treat NIC was laminectomy, an invasive surgery that removed the lamina of the spine. As laminectomy is more invasive with longer postoperative recovery, X-STOP, a
minimally invasive interspinous process implant, was introduced to treat
neurogenic intermittent claudication secondary to lumbar stenosis. This study
assesses the outcome of patients treated with X-STOP up to the 4 year
post-operative period.
Methods Used: IRB approval was obtained for this retrospective cohort
study, in which the Zurich Claudication Questionnaire (ZCQ) via telephone
was administered to an initial total of 23 patients who were implanted with
X-STOP to assess their outcome measures in three domains: 1) Symptom
Severity, 2) Physical Function, 3) Patient Satisfaction. The secondary outcome measure of the Visual Analog Scale (VAS) was used to assess postoperative trends in pain in X-STOP patients. Both the ZCQ and VAS scores
were obtained in 18 patients at the 3 year post-op period and 5 patients at
the 4-year post-op period. Five patients were lost in the follow-up process
due to unavailability or deceased status.
Summary of Results: Based on the ZCQ and VAS scores, there was a
68% (15/22) success rate following the procedure. The VAS levels at three
years were comparable to the one-year post-operative values as the pain
levels stabilized over time. For the patients reaching the 4-year post operation period, ZCQ and VAS scores remained significantly low. There were
no postoperative fractures of the spinous processes, implant dislodgement,
or wound complications.
Conclusions: X-STOP is a safe and effective treatment for NIC that provides marked relief of symptoms with sustained beneficial outcomes at
4 years of follow-up.
173
LIGHTS, CAMERA, SURGERY 2010: EXPANSION AND
STANDARDIZATION OF THE PRODUCTION OF SURGICAL
EDUCATION LEARNING RESOURCES
Kwan K, Duffy D, Masterson J, Blair G University of British Columbia,
Vancouver, BC, Canada.
Purpose of Study: Lights, Camera, Surgery (LCS) was a pilot project
launched in 2009 as part of an ongoing initiative for the development of
novel online resources aimed at preparing medical students for their clinical years. In short, the LCS project created instructional videos for teaching basic surgical procedures. The LCS pilot project was received positively
by the student participants and surgical mentors. It received competitive
grant funding for expansion and continuation of the project. The growth
of the project required implementation of a standardized format for video
production to increase efficiency, ensure consistency of content, and decrease
costs.
Methods Used: Eleven student participants were initially mentored by a
professional videographer. Surgical staff mentors coached students through
the production of the video. At the end of the project, both the student
participants and surgical mentors were surveyed using a 5-step Likert scale.
This scale was used to assess the level of engagement and learning of both
the student participants and surgical mentors in the creation of instructional
videos.
Summary of Results: In LCS 2010, two new pediatric surgical disciplines, Pediatric Cardiac Surgery and Pediatric Orthopedic Surgery, were
added to the existing five disciplines. Furthermore, two new partnerships
were formed - one with the BC Patient Safety and Quality Council, and the
other with Providence Health Care. A standardized format was created to
include 7 important aspects of a surgical procedure: 1) Outline of the Procedure 2) Indications/Contraindications, 3) Preparation, 4) Equipment, 5) The
Procedure, 6) Complications, and 7) Follow-up. Twenty-four professional
instructional videos for teaching basic surgical procedures were produced
using this new standardized format - a 26% increase compared to the pilot
project. Impressions from the student participants and surgical mentors were
obtained.
Conclusions: The implementation of a standardized format in the creation
of instructional videos for teaching basic surgical procedures was a contributing factor to an increase in efficiency during video development and
to a reduction in video production costs. The positive response to LCS
indicates that it can be a feasible and effective learning/teaching undertaking
for many procedurally-based disciplines.
174
X-STOP IMPLANTATION FOR THE TREATMENT
OF NEUROGENIC INTERMITTENT CLAUDICATION
Heyrani N1, Huo SK1, Shamie A1,2 1David Geffen School of Medicine
at UCLA, Los Angeles, CA and 2Santa Monica-UCLA Hospital, Santa
Monica, CA.
* 2011 The American Federation for Medical Research
175
COMPARISON OF RADIATION EXPOSURE AND
COST BETWEEN PERCUTANEOUS RADIOFREQUENCY
AND CRYOABLATION IN THE TREATMENT OF SMALL
RENAL MASSES
Schroeder G1, Richards G1, Agarwal G1, Smith J2, Baldwin D1 1Loma Linda
University School of Medicine, Loma Linda, CA and 2Loma Linda
University School of Medicine, Loma Linda, CA.
Purpose of Study: Percutaneous radiofrequency ablation (PRA) and percutaneous cryoablation (PCA) are being increasingly utilized in the treatment of small renal masses, but no previous study has characterized the
radiation exposure received with these modalities. The purpose of this
study was to compare radiation exposure, cost, and outcomes between PRA
and PCA.
Methods Used: A retrospective review was performed between January
2004 and April 2010 of patients with solitary renal tumors e 4.0 cm treated
with either percutaneous ablative technique. The procedure time, radiation
exposure, and cost were compared using the Mann-Whitney U Test and
Pearson Chi-Square with significance considered at p G 0.05.
Summary of Results: Fifty-three patients with small renal masses were
treated with a percutaneous ablative therapy (23 PRA and 30 PCA). The
tumor size, ASA score, and patient age were similar between groups. The
billed charges was similar between PCA ($18,079) and PRA ($17,148);
p=0.7. PCA had longer procedural time (88.2 vs. 58.6 min; pG0.001), greater
number of probes (3.2 vs 1.3; pG0.001), more CT scans (16.2 vs 10.5;
p=0.003), and greater total radiation exposure (2388 mGy-cm vs 1461
mGy-cm; p=0.02) compared to PRA.
Conclusions: No prior study has attempted to quantify the radiation
dose received during PRA and PCA. Our study demonstrates that all patients
received relatively large radiation dose during percutaneous ablation with
PCA patients receiving the largest dosage. Physicians must consider this
factor when selecting treatment modalities for young patients with renal
masses and continue to work to reduce the radiation exposure received by
these patients.
137
Western Regional Meeting Abstracts
Journal of Investigative Medicine
176
RELATIONSHIP OF ESOPHAGEAL MUCOSAL OXYGEN
SATURATION TO GASTROINTESTINAL FUNCTION
IN PATIENTS UNDERGOING CARDIOPULMONARY BYPASS:
A PRELIMINARY REPORT
Blair BM, King NC, Applegate RL, Sanghvi C, Dorotta IR, Gatling JW
Loma Linda University Medical Center, Loma Linda, CA.
Purpose of Study: Visible light spectroscopy (VLS) allows measuring of
esophageal mucosal oxygen saturation (StO2) during surgery, and reflects
perfusion of the lower esophagus. This may provide information regarding
GI tract perfusion during surgery since blood supply is shared from the
splanchnic circulation. Changes in GI perfusion may be associated with delayed recovery of normal GI function and longer hospital stay. We evaluated
the use of esophageal StO2 in adults requiring cardiopulmonary bypass (CPB).
Methods Used: This is an ongoing IRB approved observational study in
adult patients undergoing elective cardiac surgery requiring CPB. Continuous monitoring of esophageal StO2 (T-Stat, provided as an unrestricted
material grant from Spectros, Portola Valley, CA) was used. Data was captured electronically. Analysis included Pearson correlation between average
esophageal StO2 pre-, during, and post-CPB, the duration of CPB, events
associated with time on CPB, return of normal GI function/bowel movement,
and length of stay. Two-tailed significance was set at 0.05.
Summary of Results: Nine subjects have been analyzed. The magnitude
of the difference between average esophageal StO2 pre-CPB to post-CPB correlated to delayed return of GI function (r = 0.808; p = 0.008) and to length of
stay (r = 0.708; p = 0.033). In addition, esophageal StO2 levels decreased below
pre-CPB baseline levels in the transition periods on and off CPB, but neither of
these findings were statistically related to return of bowel function.
Conclusions: This preliminary analysis revealed esophageal StO2 monitoring has some limitations. Non-physiologic fluctuations in oxygen saturation levels can occur when a TEE probe is used, so measuring esophageal
StO2 during TEE needs to be explored. Our results indicate that return of
normal GI function is delayed in subjects in whom the average esophageal
StO2 post-CPB is significantly higher than the average esophageal StO2
pre-CPB. This translates into longer hospital stays, as patient discharge is
delayed until bowel function returns. We hypothesize that in this subset,
higher average esophageal StO2 post-CPB compared to pre-CPB reflects
hyperemia, which could lead to reperfusion injury of the GI mucosa.
Western Student Medical Research Forum
Student Session I - Global Health
8:30 AM
Friday, January 28, 2011
177
EVALUATION OF THE FEASIBILITY AND EFFECTIVENESS
OF A COMPLEMENTARY FEEDING INTERVENTION FOR
REDUCTION OF CHILDHOOD MALNUTRITION IN KENYA
Ashton R2, McCalmont K2, Tomedi A1, Rohan-Minjares F1 1University of
New Mexico School of Medicine, Albuquerque, NM and 2University of
New Mexico School of Medicine, Albuquerque, NM.
Purpose of Study: To study the operational feasibility and effectiveness
of the distribution of locally-available foods to prevent malnutrition and
improve child growth in Kenyan children.
Methods Used: A quasi-experimental design was chosen with an intervention group consisting of children in all of the villages in one region of
arid, rural, Eastern Kenya, and a non-intervention comparison group of
children in all of the villages in an adjacent region. Children included in the
study were age 6 to 20 months with weight-for-height z score 9 j2 at baseline. The intervention was the distribution of a monthly food ration for the
index child and separate ration for the family, and group education on appropriate complementary feeding and hygiene. At baseline and at the end
of the 7-month intervention, community health workers visited every household to measure child length/height and weight. Each month a questionnaire
was administered to the intervention household to assess receipt of supplemental food and degree to which the index child received the food.
Summary of Results: Children in the intervention (n = 141) and control
(n = 185) groups had similar baseline anthropomorphic measures. The
138
&
Volume 59, Number 1, January 2011
caretakers in the intervention group confirmed that the intended amounts
of food supplements were received monthly, and child nutrient intake was
significantly greater in the intervention group. During the 7-month intervention, the growth of the children, measured by mean z score, was significantly better in the intervention group than the control group, weight for
age difference = 0.79 (p G .0001), weight for height difference = 1.17
(p G.0001). Compared to the control group, the intervention group had a
lower prevalence of wasting (0% vs. 8.8%, p = .0002) and underweight
(5.4% vs. 22.4%, p G .0001). Infectious morbidity (diarrhea and respiratory
infections) was similar in both groups.
Conclusions: The findings suggest that the distribution of locally available
foods is operationally feasible and improves child growth and decreases
malnutrition in Kenyan children.
178
NUTRITION EDUCATION IN THE BHUTANESE NEPALI
REFUGEE COMMUNITY OF TWIN FALLS, ID
Brito T University of Washington, Seattle, WA.
Purpose of Study: The College of Southern Idaho Refugee Program
(CSIRP) has resettled 2,500 refugees since 1980, empowering them to live
successfully and independently in Twin Falls. In the last 2 years, many new
arrivals have been Bhutanese Nepali. Post-migration diets of refugees tend
to be heavily dependent on refined sugars, polished grains and nutrient-poor
snacks. A presentation was designed to educate about making choices that
support a balanced diet and healthy lifestyle.
Methods Used: Interviews of CSIRP staff members identified nutrition as
a challenge for refugees. Review of the primary literature confirmed the
effectiveness of a visually supported intervention based on the USDA Food
Pyramid guidelines for an audience with varying degrees of English proficiency. Attendance at a Bhutanese Nepali festival provided cultural context
for traditional diet. A poster based on the Food Pyramid with photographs
illustrating each food group was created as part of a presentation for an ESL
class; an interpreter was asked to attend.
Summary of Results: Twenty-five refugees attended the presentation
during an ESL class. Though an interpreter was available, most attendees
could follow the presentation in English with the aid of the poster. The target
audience was engaged and asked clarifying questions about adapting traditional recipes. It sparked a discussion about the consequences of poor
nutrition choices made by fellow refugees. Many asked for and were referred
to the USDA website in order to create a personalized diet plan.
Conclusions: The presentation was well received; the audience was involved and interested as evidenced by the relevant questions and discussion.
Translation by the interpreter was not necessary for audience comprehension; this was a success as the intent was for basic language and symbols
on the visual to convey the message adequately to those without a mastery
of English. The poster and presentation will be included as a part of future
ESL classes. The refugee community in Twin Falls is very close knit and it is
common for acquired knowledge to be passed on from old to new refugees
as they are welcomed to their new life. Hopefully the information from this
presentation can be part of that exchange. Though educational interventions
aimed at a non-English-speaking population can seem intimidating, they are
possible and often some of the most necessary.
179
ADDRESSING THE HEALTHCARE WORKER SHORTAGE
IN RURAL UGANDA BY TRAINING VILLAGE
HEALTH WORKERS
Cluka B University of Washington, Seattle, WA.
Purpose of Study: Uganda is currently suffering from a severe deficit
in healthcare workers, with only 81 medical personnel for every 100,000
people. The rural areas are disproportionately impacted by this shortage.
In an effort to address this shortage, Uganda has recently turned its attention toward task shifting, which is the transfer of certain responsibilities
from a doctor or nurse to a healthcare worker with less training. A cornerstone of this concept is the training of Village Health Workers (VHW_s).
This project took place in Kiboga, a rural district in central Uganda with
a population of 300,000 and a mere five physicians responsible for direct
patient care. A VHW training program was conducted in collaboration with
the NGO Global Youth Partnership for Africa (GYPA).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Methods Used: A group of 19 volunteers attended courses four hours
Purpose of Study: 56% of all childhood deaths due to infectious diseases
per week for a total of six weeks. Lectures were based on David Werner_s
BWhere There is No Doctor[ and students were supplied with a copy of
this book and a first aid kit. The author interviewed district officials to ascertain what skills the village health workers needed, and created and
taught classroom sessions in collaboration with a GYPA volunteer physician. A pocket guide was designed by the author and distributed to the
students. Additional copies of the pocket guide were provided to previous
village health worker graduates and to the program coordinator for use in
future courses.
Summary of Results: 18 of the 19 volunteers successfully completed
the six-week training course. The students were enthusiastic about the
training, as they felt it would prepare them to take an active role in influencing the health of their community. Residents throughout Kiboga who were
interviewed stated that the implementation of village health teams has contributed to improved health outcomes and will continue to have a positive impact
on the community. GYPA will continue to organize the course and train
volunteers.
Conclusions: Training VHW_s can help combat the healthcare worker
shortage and leads to improved health literacy throughout the community.
Further analysis is needed in order to ascertain whether training and utilizing VHW_s results in measurable improvements in health outcomes. Additional work is necessary in order to increase collaboration between the
district and the NGO_s teaching the courses.
are associated with underlying malnutrition of which vitamin A deficiency
(VAD) is the most prominent, accounting globally for 667,771 deaths
per year, or 6.5% of all deaths under five. VAD in Uganda is reported in
52% of women and 32% of children, despite national supplementation
campaigns. Since VAD co-presents with infectious diseases and supplementation is shown to improve disease outcomes, the integration of supplementation with routine care serves to amplify both interventions.
Methods Used: A literature review of the indications for vitamin A supplementation (VAS) with the management of malaria, diarrhea, measles,
pneumonia, and HIV in children under 5 was performed and distilled into a
simplified flowchart. The flowcharts were distributed to the entire staff at
Kiboga District Hospital during a CME presentation about VAS. Follow up
training of healthcare providers in the outpatient, maternity, and pediatric
wards was provided to promote the adoption of VAS into regular clinical
practice. Additional work with the pharmacy was done to procure a steady
supply of vitamin A capsules to guarantee sustainability of the practice.
Summary of Results: At the initial CME training, 33 hospital staff
members were instructed about the indications for VAS. Further training
of 12 doctors, nurses, and clinical officers integrated VAS with routine visits
in the outpatient ward and daily nursing rounds in the pediatric and maternity wards. Training also stopped the practice of VAS in HIV positive
postpartum mothers since it increased the risk of MTCT through breastfeeding. Coordination with the head pharmacist secured 20,000 capsules
for immediate use in the project with the prospects of obtaining additional
capsules from the MOH National Stockpile.
Conclusions: The project achieved its goal of integrating the cost-effective
intervention of VAS with the routine clinical management of infectious
diseases in a setting with a massive VAD public health problem. The additional work with the supply of vitamin A capsules ensures sustainability of
the project for the immediate future. However, additional progress in food
fortification, diet diversification, and poverty reduction will need to be
achieved in order to directly address the underlying causes of VAD.
180
REDUCING TYPHOID INFECTIONS IN ANTSIRABE,
MADAGASCAR THROUGH THE PROMOTION
OF HANDWASHING AND HOUSELHOLD
WATER PURIFICATION
Embick E University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Typhoid fever is very common in Madagascar. At
Andranomadio Lutheran Hospital (HLA), located in the central highland
town of Antsirabe, it was of the most common reasons for hospitalization.
During the months of December - July, 2010, the rates of hospitalization
for typhoid fever increased dramatically. Approximately 1/3rd of pediatric inpatients between June 14th and July 10th had been diagnosed with the disease. Since typhoid is transmitted through fecally-contaminated food and
water, an education program was created to promote hand washing and
point-of-use water treatment in an effort to decrease the spread of the disease.
Methods Used: Typhoid was identified as a major health issue in Antsirabe
during a community health assessment. An educational program was created
based on information gathered from the scientific literature and discussions
with local healthcare providers, health-related NGOs, and the families of
typhoid patients. A presentation was given, discussing ceramic filtration,
chlorination, solar disinfection, and boiling as water purification methods, and
a local NGO was identified which produces low-cost water chlorination solution for household use. The presentation also reviewed methods of safe
water storage, as well as standard hand washing technique using soap. A
Malagasy-language, picture-based handout covering key points was created
and distributed.
Summary of Results: The presentation was given to a group of approximately 150 people at a Lutheran Church conference on the outskirts of
Antsirabe. One hundred brochures were distributed to attendees afterward.
The presentation was about twenty minutes long and was given in French
and translated into Malagasy for the congregation.
Two-hundred brochures were left for three pediatric nurses, who agreed
to discuss typhoid prevention with their patients and hand out the brochures.
Conclusions: This project met the goals of helping to increase awareness
of typhoid fever and educating the population about simple affordable
methods to prevent the disease. Additional educational efforts are warranted
in order to reinforce these messages and promote preventative behaviors in
a larger portion of the local population.
181
REDUCING CHILD MORTALITY THROUGH THE
INTEGRATION OF VITAMIN A SUPPLEMENTATION WITH
THE MANAGEMENT OF INFECTIOUS DISEASE IN UGANDA
Furukawa N University of Washington, Seattle, WA.
* 2011 The American Federation for Medical Research
182
COMMUNITY-BASED DENTAL HEALTH EDUCATION
IN THE PHILIPPINES
Hale NA University of Washington, Seattle, WA.
Purpose of Study: Dental decay is one of the most common childhood
illnesses experienced in the Philippines. Upwards of 95% of Filipino children are affected and toothache is often cited as the most common reason
for school absenteeism. To address this issue at Gawad Kalinga (GK)
Santo Niño, a collaborative pediatric dental health education program was
established.
Methods Used: An integrative dental health program, the Bright Smiles
Club (BSC), was formed for children ages seven to thirteen as part of the
preexisting SAGIP youth group. Three local dentists volunteered to conduct
a screening clinic, train volunteers, and provide follow-up care for those
with the worst periodontal disease. Toothbrushes and fluoride toothpaste
were donated by Colgate Philippines. Eight healthcare volunteers (HCV_s)
led two educational sessions with the BSC on improving dental health after
receiving training from the author. As part of their participation, the children created posters on nutrition and dental health and learned the Colgate
educational song BEveryone Has a Smile.[ Parents were debriefed and received Tagalog-language dental hygiene pamphlets from the Department
of Health. In addition, a dental health tracking sheet was created for students to log daily habits, including tooth brushing; the consumption of
fruits, vegetables, and milk; and the avoidance of soft drinks and candy.
A monthly prize was awarded to the student with the highest number of
points. To ensure program sustainability, a continuity plan was distributed to
the HCV’s.
Summary of Results: 31 children participated in the program. Dental
screening revealed significant disease: 28 were found to have one or more
caries and extraction of an average of three teeth per child was recommended for 25 participants. The children enthusiastically participated in the
program and many drastically modified their health habits. The BSC received
an overwhelmingly positive response from the community and HCV_s commented that children_s teeth seemed healthier and that participants were demanding nutrient-rich foods.
Conclusions: This project illustrated that multifaceted, community-based
educational programs can be effective in improving oral health, and can serve
139
Western Regional Meeting Abstracts
Journal of Investigative Medicine
as a framework for other health-promotion activities. Future community
health projects should focus on creatively encouraging increased parental
participation.
183
REDUCING INTESTINAL PARASITIC INFECTIONS
IN RURAL PERU THROUGH COMMUNITY EDUCATION
Pudwill L University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Helminth infections are the second-leading cause
of illness in Yantaló, a village in the tropical Amazon of northern Peru,
for 2006Y2010. The population is susceptible to parasitic infections due to
the unsafe water, poor sanitation, crowded conditions, low education levels,
poverty, agricultural activity, and subtropical climate. The aim of this project was to reduce intestinal parasitic infections in Yantaló through sustainable community-based education.
Methods Used: High school students were taught about parasite life
cycles, health consequences, and prevention. Student volunteers then developed and gave presentations about hygiene and parasites to younger
children. A presentation was given to a group of adults and hand-washing
activities were done with kindergarten students. Educational materials were
prepared for future use by educators, health workers, and volunteers with
the Yantaló Foundation. The education coincided with the village_s first
anti-parasite medication distribution campaign, sponsored by the Ministry
of Health. The work was done with the help of volunteers from the Yantaló
Foundation.
Summary of Results: A total of 214 high school students received education about parasite prevention. Six groups of high school students gave
presentations to 248 elementary students in twelve classes. Twenty adults
attended the lecture about parasite prevention and received a handout. Fifty
kindergarten students participated in the hand-washing sessions. All educational materials and information were given to the Foundation for future
use in the community. The school nurse agreed to supervise high school
students in teaching younger children. The clinic physician was assisted in
the distribution of Albendazole to school children, which will be repeated
every three months.
Conclusions: The project increased awareness of hygiene, water treatment, and parasite prevention among school children through education
that promoted community involvement. The education will be sustainable
with cooperation of local students, the Yantaló Foundation, and the school
nurse. It will be important to complement education with periodic distribution of anti-parasite medication. Additional adult education and water treatment measures are also necessary to reduce the prevalence of parasitic
infections in Yantaló.
184
WHAT NEW KNOWLEDGE DO STUDENTS GAIN
FROM INTERNATIONAL HEALTH WORK?
To F University of British Columbia, Vancouver, BC, Canada.
Purpose of Study: To determine if students gain new knowledge in global
health work after participation in a student-driven international project.
Methods Used: The 17 UBC students that participated in the Global Health
Initiative (GHI) projects were surveyed. Students were asked to rate their
pre and post project knowledge in four areas: cross cultural communication,
project development, project sustainability, and community collaboration/
local empowerment. Mean relative percentage change in each learning area
was calculated. The survey also collected qualitative information on project
learning outcomes by asking BWhat were the most valuable lessons you
learned while working overseas?[.
Summary of Results: Quantitative Results: Students consistently reported
that they increased their knowledge in each of the measured learning areas.
Mean relative increase in each area: cross-cultural communication 44%,
project development 119%, project sustainability 111%, and community
collaboration/local empowerment 105%. Qualitative Results: Even though
there was diversity in participant responses, common lessons and themes
emerged. These include the value of appreciating cultural norms and work
ethics, relationship building with locals, interdisciplinary collaboration, effective communication practices, flexibility in project implementation, thorough
pre-departure preparation (project planning and cultural awareness), and development of locally-sustainable projects.
140
&
Volume 59, Number 1, January 2011
Conclusions: Through partaking in GHI projects, students consistently
demonstrated significant increases in their understanding of international
health project development based on the four learning objectives measured
in this study. Students also have garnered many other lessons in international
health from participation in GHI as reported in the qualitative results.
185
THE TREATMENT AND PREVENTION OF DIARRHEA
AND DEHYDRATION IN RURAL MONGOLIA
Wallace R University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Diarrhea is pervasive in the rural Darhad Valley of
Mongolia due to contamination of surface waters coupled with inadequate
sanitation measures. With no access to oral rehydration packets and little
knowledge concerning rehydration therapy in the community, individuals
with diarrhea, especially children, are at risk for dehydration and death.
Therefore, an educational program was established to instruct community
members about the causes, treatment, and prevention of diarrhea and
dehydration.
Methods Used: Based on a review of recent literature a presentation was
designed to address diarrhea and dehydration in bagh (township) 1 in the
Darhad Valley. With the organizational help of community leaders, the presentation was given to multiple sites within the region including an annual
community festival called Nadaam that attracted much of the population.
To complement the presentation, a muslin poster was made that utilized
pictures and Mongolian text. Additionally, to educate local physicians, a
laminated pamphlet was made with BioRegions International, an NGO with
12 years experience in the area, addressing dehydration and rehydration
therapy. Lastly, a presentation was made for the Darhad Valley Kindergarten
class regarding hand washing and other basic sanitation measures.
Summary of Results: The diarrhea/dehydration presentations were given
to over 50 members of the community; approximately half of the 80 families
in bagh 1 were present. Furthermore, the poster and the notes for the presentation have been left with BioRegions International for future use. The
dehydration pamphlets were distributed to over twenty doctors in the region
during an annual seminar. The handouts were well received and have been
observed hanging on display for the community in several clinics. Lastly,
around 25 children and some of the parents attended the Kindergarten
presentation.
Conclusions: The project successfully met the goals of increasing awareness among the community concerning the causes of diarrhea, the importance of hygiene, ways to identify dehydration in infants and adults, and
the treatment of dehydration using homemade oral rehydration salts. Additional efforts to combat diarrhea are needed, such as improving the quality of
surface waters, promoting zinc tablets, and instituting rotavirus vaccination
programs.
186
PEDIATRIC MALARIA TREATMENT DELAYS ACROSS
SOCIOECONOMIC LEVELS IN SUBURBAN ACCRA, GHANA
Wong KR 1Yale University School of Medicine, New Haven, CT; 2University
of California at San Diego, La Jolla, CA and 3University of Ghana School
of Public Health, Accra, Ghana.
Purpose of Study: The study reported here examines how a Ghanaian
family_s socioeconomic status (SES) affects the delay-time before a child
with Plasmodium falciparum malaria is brought to a government hospital.
It was hypothesized that a delay of appropriate clinical treatment results from
a family_s apprehension of inability to pay the hospital fee.
Methods Used: In this ethnographic study, parents of pediatric patients with
blood smear-confirmed malaria (N=15) were interviewed through both
quantitative questionnaires and narratives at the Legon Hospital Pediatrics
Ward. The questionnaires ascertained the dependent variable (time lag during
which the child languished at home), as well as the independent variable
of SES, measured by: 1) monthly income range, 2) health insurance, and
3) years of household education. Over the course of four months, 170 hours
of dialogue and observation supplemented the numerical data.
Summary of Results: There was actually no statistical association between
delay-time and family financial standing, nor a correlation between delaytime and enrollment in Ghana_s recently-established National Health Insurance Scheme. Parents of all backgrounds postpone clinical treatment even
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
after the onset of clear malaria symptoms in order to conserve their time;
the waiting lines at hospitals are often several hours long and the illness
is endemic in West Africa. Yet, this study does confirm that lower-SES
households less effectively prevent malaria. Only half of families in the
lowest income group utilized insecticide-treated mosquito nets, while all
six families in the highest income group use the precaution.
Conclusions: In this township, the study suggests that low-SES families
do not disproportionately delay clinical treatment. Especially when raising
multiple children, parents of all SES groups delay at home and self-treat
with herbal teas or street-bought (and often counterfeit) chloroquine and
artesunate anti-malarials. This has its risks. Delayed clinical care and the
rapid deterioration of originally-uncomplicated P. falciparum cases contribute to 20,000 pediatric malaria fatalities in Ghana annually.
187
FACTORS AFFECTING PERCEIVED HEALTH OF LATINA
CAREGIVERS IN EAST LOS ANGELES
Wolchok LM1, Mendez-Luck C2 1UCLA, Los Angeles, CA and 2UCLA, Los
Angeles, CA.
Purpose of Study: To examine the effect of perceived burden on the
health of female Mexican and Mexican-American caregivers who give unpaid
assistance to elderly relatives. Currently, eight million Latino adults provide
informal, unpaid care to adult relatives, and the population of Latino caregivers is likely to continue to grow with the growth of the older Latino
population. In order to better support these caregivers, it is important to
understand their caregiving experiences.
Methods Used: We analyzed data from a convenience sample of caregivers (N = 96) on different domains of caregiving, including perceived
physical and emotional intensity, number of ADLs/IADLs, and cultural
reasons for caregiving. We conducted univariate, bivariate, and multivariate
analyses to understand the caregiver population demographics, characteristics of the caregiving situation, attitudes toward caregiving, and how these
factors affect caregiver self-perceived health. Analyses were done using
PASW Statistics GradPack Version 18.0.
Summary of Results: Better caregiver health was associated with higher
education, younger caregiver age, fewer forms of support provided to the
care recipient, the care recipient not having dementia or mental illness, and
lower perceived physical intensity of care. However, only perceived physical
intensity of care was significant in our final regression model.
Conclusions: These results suggest that subjective measures of caregiver
burden like perceived intensity are stronger predictors of Latino caregivers_
health than are objective measures of the amount of care provided.
Western Regional Meeting Abstracts
to pursue further program development. Cooperation was made with the
local hospital_s Physical Therapy department.
Summary of Results: Involving future participants in the initial program
design and addressing their concerns at the forefront was essential to success. Identified infrastructure concerns relating to sidewalks, crosswalks,
and facilities, were directed to the city council. Proposals were made to
involve area youth programs in cleaning a local walking trail. Interest and
perceived benefits of the program were demonstrated by a 224% increase in
attendance from discussion group (49) to seminar attendance (110). Forty
Bpractical exercise[ packets were distributed. A report summarizing the research and senior concerns was distributed to local city and healthcare
leadership to guide future program development.
Conclusions: The benefits of increasing physical activity for seniors are
significant. The success rate of senior fitness programs is greatly improved
by establishing a community-wide approach. Programs are best initiated
after surveying for concerns of the participants, tailoring the program to
address those concerns, and then educating participants prior to program
initiation. Community interest and education will be enhanced.
189
AN ALTERNATIVE SAFETY NET TO THE EMERGENCY
ROOM: A FREE MEDICAL CLINIC FOR THE HOMELESS
IN SPOKANE, WASHINGTON
Western Student Medical Research Forum
Student Session II - Community Health
8:30 AM
Friday, January 28, 2011
Inaba CS University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Community free clinics offer healthcare services to a
growing disenfranchised population that might otherwise use the emergency
department for costly treatment of nonurgent medical issues. Using as a
model the House of Charity (HOC) Outreach Clinic in Spokane, Washington,
this study seeks to explore how a local free clinic can prevent unnecessary
ER visits by providing free outpatient care to the underserved.
Methods Used: Patients seeking free medical care at the HOC were surveyed by questionnaire and chart review. Data collected include where
patients would seek care if the HOC were unavailable, clinic and ER usage,
health status, housing status, insurance coverage, sociodemographics, chronic
illnesses, and reasons for visit and treatments received at the HOC.
Summary of Results: Preliminary results based on 26 surveys collected
between June and August 2010 indicated that 56% of respondents would
wait to receive treatment at the HOC free clinic instead of using the emergency department (28%) if the HOC were temporarily unavailable. 50%
versus 31% indicated that they use the HOC clinic instead of the ER, respectively, as their primary source of medical care. 65% had no insurance
and 50% had been homeless for over six months. Reasons for visit included
primarily medication refills for chronic medical conditions.
Conclusions: The HOC Outreach Clinic serves as a primary source
of medical care for disenfranchised individuals who might otherwise seek
expensive ER treatment for chronic health issues that can be addressed in the
outpatient setting. Funding used to cover the high cost of nonurgent hospital
treatment might be better applied toward the support of free clinics that
provide similar treatment at lesser cost.
188
DEVELOPING PHYSICAL FITNESS PROGRAMS
FOR SENIOR ADULTS IN MONTPELIER, ID
190
SOCIODEMOGRAPHIC PREDICTORS OF TRAVEL
DISTANCE TO A FREE CLINIC IN LOS ANGELES
Aebischer TD UW School of Medicine, Seattle, WA.
Purpose of Study: Diabetes, hypertension, coronary heart disease, and
other Chronic Health Conditions (CHCs), affect over 100 million Americans,
with 88% of those over 65 having at least one or more CHC. Moderate
physical activity provides a 30% reduction in incidence of CHCs. This project
focused on establishing a model for designing and initiating a senior adult
focused physical fitness program that encourages initial participation and
long-term program adherence, a model to be used in Montpelier and other
similar rural communities.
Methods Used: Surveys involving area healthcare providers and leaders
identified the absence of fitness programs. A literature review identified
benefits and best practices for designing and initiating senior oriented fitness
programs. The local newspaper provided project awareness. Area seniors
were gathered for a discussion group focusing on their ideas regarding
attractors and detractors related to physical fitness. An educational seminar
was held covering benefits of physical activity and providing specific exercises addressing identified physical concerns. Local leaders were engaged
Kim MJ1, Ra H3, Han YK1, Jo A2 1David Geffen School of Medicine at
UCLA, Los Angeles, CA; 2David Geffen School of Medicine at UCLA, Los
Angeles, CA and 3College of Letters and Science, UCLA, Los Angeles, CA.
Purpose of Study: Patients often seek health services from free clinics,
sometimes traveling inconvenient distances to do so. We sought to understand the relationship between travel distance and various sociodemographic
factors among patients accessing the UCLA-Korean Resource Center Community Health Center (UK-CHC).
Methods Used: Chart reviews were conducted on 145 patients seen at
UK-CHC from August 2008 to March 2010. Clinic staff calculated individual patients_ travel distance from place of residence to the clinic using Google
Maps. Using STATA 9.1 software, we examined the relationship between
distance traveled and the following variables: age, gender, insurance status,
employment status, language preference, education, and proportion of life spent
in the U.S.
Summary of Results: On average, patients traveled 11.4 miles to reach
the clinic (range: 0Y73.8 miles). The average length of U.S. residence was
* 2011 The American Federation for Medical Research
141
Western Regional Meeting Abstracts
Journal of Investigative Medicine
15.1 years, or approximately 30% of their lifetime. Almost all of the
patients preferred to speak in the Korean language, about half (54%) were
not working and a great majority (92%) reported no health insurance. More
than half (58%) reported having received college education or higher. In
our multivariate analysis, we found that patients lacking health insurance
traveled an average of 10.99 miles farther than their insured counterparts.
Age was weakly inversely associated with the distance traveled.
Conclusions: Among the variables examined, lack of health insurance
was the strongest predictor of the distance of travel for patients in seeking
health services at UK-CHC. This suggests the importance of taking travel
distance into consideration when referring patients to other sites that can
provide more comprehensive health services. Additionally, our results
point to the pressing need for the expansion of health insurance to those
who lack it.
191
COLLABORATION IN SUBSTANCE ABUSE PREVENTION
Marvinsmith BD University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Within the Confederated Salish and Kootenai Tribes
(CSKT) of Montana, there is a significant need for youth substance abuse
prevention (SAP) programs. Tribal Health and Human Services (THHS) is
a comprehensive network of health care providers available to the CSKT
community. Despite a variety of social programs managed by the Tribes,
no youth SAP program exists. The purpose of this project was to provide
new information and foster collaboration in SAP among the care providers
of the tribal community.
Methods Used: Consultation and observation with the primary care providers and behavioral health staff members of THHS demonstrated the
need for SAP in the community. All providers confirmed little collaboration between primary care and behavioral health. A literature review was
performed to garner new information on SAP in tribal communities, and a
presentation on these strategies was created. All studies examined were
published within the past two years (since 2008). The presentation was given
at a regularly scheduled meeting for the clinicians of THHS. The behavioral
health staff was invited to attend.
Summary of Results: The literature revealed new efforts in SAP in tribal
communities. The most successful were in Community-Based Participatory
Research (CBPR). Prevention strategies, evidence-based practice guidelines,
and a detailed explanation of CBPR were presented. A list of research and
academic institutions focused on SAP work with tribal communities was
included. A reference list of strategies reviewed, including point of contact,
was provided. Ten staff members attended the presentation, including representatives from clinicians, behavioral health, and administration. A twentyminute discussion among the providers followed the presentation. All attendees
received a handout. The director of behavioral health requested an electronic
copy of the presentation.
Conclusions: The providers of THHS are a crucial rallying point in development of a youth SAP program due to their interaction with patients
as well as recognition from the relevant authorities within the Tribes. This
educational presentation provided a forum for inter-professional collaboration as well as a basis for developing an indispensable program.
192
ADDRESSING POSTPARTUM TESTING OF GESTATIONAL
DIABETES MELLITUS IN OTHELLO, WA
Morgan K University of Washington School of Medicine, Seattle, WA.
Purpose of Study: The incidence of gestational diabetes mellitus (GDM),
or diabetes affecting pregnant women, is on the rise in the US. The Columbia
Basin Health Association including the Othello Family Clinic sees a large
population of pregnant women, the majority of which are Hispanic. Minority
populations have a higher risk of developing postpartum diabetes if they
have been diagnosed with GDM. Yet there is no system in place to remind
providers to perform postpartum retesting and as a result many women are
not retested. The purpose of this project was to increase provider and staff
awareness of postpartum GDM screening guidelines and to discuss ways to
increase the number of women retested for GDM at their postpartum visit.
Methods Used: Discussions with staff and observation of patient encounters were utilized to determine the health issues in Othello and the patient
group that was to be the focus of the project (recently postpartum women
142
&
Volume 59, Number 1, January 2011
diagnosed with GDM). Clinic quality managers provided clinic statistics
on the number of women diagnosed with GDM in 2009, the number of these
women retested postpartum, and the number of GDM women that developed
postpartum diabetes. A literature review was performed to find the current
guidelines for postpartum GDM testing and to find studies in which solutions
to low postpartum testing were assessed. A presentation was given at the
monthly High Risk OB meeting to increase the awareness of postpartum
GDM testing guidelines and the clinic_s current state of retesting.
Summary of Results: A presentation on the postpartum testing of GDM
was given to a group of twenty-three CBHA providers, hospital nurses,
maternal services staff, and case managers at a monthly High Risk OB
meeting. The presentation ended with a discussion of the obstacles to
retesting and the future steps needed to increase postpartum testing numbers.
A possible system for generating reminder recalls to be sent to both providers
and patients was discussed.
Conclusions: Providers and staff understand the need to improve postpartum testing of GDM, especially with the Hispanic population they serve.
Obstacles to retesting include provider forgetfulness and lack of postpartum
follow up care. By increasing awareness of the current retesting status and
the current guidelines for postpartum testing of GDM, we can increase the
quality of postpartum care of women with GDM.
193
CREATION OF A SUPPORT GROUP FOR FAMILIES OF
DRUG AND ALCOHOL ABUSERS
Naderi R University of Washington, Seattle, WA.
Purpose of Study: Alcoholism and drug abuse are prevalent through all
age groups in the geographically isolated community of Forks, WA. Forks
and the surrounding Native American reservations experience high rates of
poverty and unemployment as well as inadequate activities for teenaged
populations. Though rehabilitation services are available for the substance
abusers, there are very few resources available for family and close friends of
abusers. The objective of this community health project is to initiate dialogue
among the population for the establishment of a support group for family
members of substance abusers.
Methods Used: A cross-disciplinary group of healthcare providers was
interviewed to identify and verify the need for a family recovery resource.
Educational materials from the only available family support program, Al-Anon,
were compiled and reproduced for more widespread use. A literature review
validated the significance of family support groups in the recovery of a substance abuser. An in-depth analysis of the nationally predominant twelve-step
recovery programs was performed. A community meeting was organized for
families and friends of drug and alcohol abusers, to discuss the possibility of
a new family support system. A publicity campaign was initiated to distribute
flyers among local businesses and at chemical dependency centers on Indian
reservations. It was arranged for Families Anonymous meetings to be held
at least for a month following this meeting.
Summary of Results: Health care providers in the area, especially those
working with patients on pain management regimens, supported the effort
by informing patients and families about the meeting. 15 people attended the
meeting and received handouts from Al-Anon, Families Anonymous, and
other 12-step family support groups. There was an open discussion about
the community_s response to the support group. Research findings were
presented, and guest speakers reinforced the effectiveness of family support
groups in the recovery of the addict.
Conclusions: People learned more about Al-Anon and other resources
available in the area. Attendees were enthusiastic about the initiation of
a Families Anonymous chapter. There was discussion of the creation of an
Al-ateen chapter for children living in homes with alcoholics. In summation,
the campaign was well embraced by the community.
194
PROMOTING CHILDREN’S ORAL HEALTH IN
SHELTON, WASHINGTON
Parker M University of Washington, Seattle, WA.
Purpose of Study: One year olds in Washington are five times as likely
to have cavities than average American one-year-olds, and other age groups
show a higher rate as well. This problem is apparent in Shelton, where, like
many of Washington_s rural towns, the water supply is not fluoridated.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Cavities are the most prevalent childhood health problem, and are preventable
with good hygiene. Pediatricians can play a major role in the prevention of
dental disease by educating parents about oral hygiene and dental visits for
children and babies. The purpose of this project was to increase families_
knowledge about preventing dental decay.
Methods Used: Clinical observations and conversations with clinicians
helped identify causes of childhood caries. A literature review was conducted
in order to find educational materials for families, as well as recommendations for counseling parents about oral hygiene and dental care. A family oral
health education table was set up at a community fair, and included interactive materials and supplies from the Department of Health. A brief presentation was given to the providers at Oakland Bay Pediatrics about the
effectiveness of oral health counseling by pediatricians, and recommended
educational points. Oral health posters in English and Spanish were ordered
and installed in exam rooms.
Summary of Results: Twenty-five children participated at the community
fair education table. Several parents said that they did not know that children
should go to the dentist at age 1, and were given educational pamphlets. The
providers at Oakland Bay were very receptive to the information presented,
and were encouraged by the evidence for the effect of parent counseling on
children_s rate of cavities. The providers agreed that more materials were
needed in exam rooms, and are considering putting a reminder in the EMR to
counsel about oral health.
Conclusions: Childhood tooth decay is a complex problem that should be
targeted through public health policy, education, and social programs promoting access to quality health care and healthy foods. Parents need help
learning about existing resources and what they can do to prevent dental
decay in their young children. The providers at Oakland Bay are concerned
about the high rate of dental disease in their patient population, and are
increasing their knowledge and resources to address this problem.
195
IMPROVING IMMUNIZATION ADHERENCE IN THE
COMMUNITY OF SHOSHONE, IDAHO
Rohrbach M University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Reports from America’s Health Rankings indicate that
Idaho ranked 49th in the nation regarding immunization coverage for children ages 19 to 35 months in 2009. Idaho recorded a 65.9 percent coverage,
falling nearly 20 percent below the nation’s top state. The focus of this
intervention was to identify barriers of adherence and initiate time-efficient
strategies for improving immunization rates within the rural community of
Shoshone, Idaho.
Methods Used: To identify the healthcare needs in the community, efforts involved questioning social workers, health educators and clinicians.
After identifying an area of concern, a literature review led to the assembly
of a vaccine information packet (VIP) designed to address the barriers
of adherence identified by recent studies. The most common barriers investigators discovered included: parents’ unanswered concerns; a lack of
education; a lack of direction; poor access to care; and various socioeconomic factors. The initial intervention site in Shoshone utilized an
established event known as the Wlunch in the parkW program, which provides free lunches for children, to serve as the introduction of VIP use.
Additional VIPs were also supplied to the nurse heading immunizations
at the Shoshone Family Medical Center in anticipation of future use, both
in clinical and public settings.
Summary of Results: During the intervention, parental attendance
approached 30 individuals and the inadequate vaccination rates in Idaho
were brought to the attention of more than a dozen parents. Parents’
questions and concerns were addressed and nine VIPs were distributed,
along with various supplemental resources regarding vaccination. Nurses
at the medical center in Shoshone plan to use VIPs in the future during
school registration, for vaccination reminders, and as needed in the medical center.
Conclusions: The state of Idaho has already taken action by supplying
providers with vaccinations free of charge, however these efforts are proving inadequate as they fail to address the majority of barriers clinicians
face. VIPs may be useful in addressing such matters, and although any influence on immunization rates cannot currently be measured, parents were
receptive to the VIPs. Change will not come overnight, but by educating,
informing and addressing parents’ questions and concerns there is hope of
improving rates over time.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
196
PSYCHOSOCIAL MEDIATORS TO PHYSICAL ACTIVITY
DURING THE PERINATAL PERIOD: A SYTEMATIC REVIEW
OF THE LITERATURE
Stevens AR, Leiferman J University of Colorado at Denver, Denver, CO.
Purpose of Study: Less than half of US women reported regular exercise
during pregnancy and 46 percent of normal weight women, 46 percent of
obese women, and 59 percent of overweight women gain in excess of the
recommended gestational weight advised by the Institute of Medicine. This
evidence suggests that a sedentary lifestyle contributes to excessive gestational weight gain, which identifies a need for effective intervention strategies
that target mediators to physical activity (PA) in order to increase activity
levels in perinatal women. The purpose of this study is to critically review
prospective and intervention studies identifying potential mediators to PA
during the perinatal period.
Methods Used: PubMed, MEDLINE, CINAHL, and PsychINFO were
searched for prospective studies that examined potential mediators to PA, as
well as any intervention studies that aimed to increased PA levels during
pregnancy up to two years postpartum. Fourteen prospective studies and 7
intervention studies were selected that met the inclusion criteria of this review. Articles were reviewed and discussed according to the population,
study design, mediators examined, measured outcomes, results, conclusions
and limitations of the studies.
Summary of Results: The prospective studies identified several significant
correlates of PA during the perinatal period including exercise and barrier
self-efficacy, social support, prepregnancy exercise behavior, safety concerns,
and perceived stress. Three intervention studies significantly increased PA
levels, three did not achieve significant increases in activity, and one study
performed an intervention to alter the potential mediators to PA, but did not
measure activity levels.
Conclusions: The most commonly reported significant predictors of PA
during the perinatal period in this review were self-efficacy and prepregnancy
exercise behavior. Several intervention studies followed theoretical frameworks and targeted various mediators to PA. However, very few mediators
were directly measured to determine the efficacy of the interventions in altering the mediators and PA levels. Further research is needed to establish
significant mediators in the perinatal period, as well as effectively alter these
mediators in intervention programs.
197
SCREENING PREGNANT WOMEN FOR ADVERSE
CHILDHOOD EXPERIENCES IN PORT
TOWNSEND, WASHINGTON
Wooldridge B University of Washington, Bremerton, WA.
Purpose of Study: To train medical staff at Olympic Primary Care (OPC)
in Port Townsend, Washington on Adverse Childhood Experiences (ACE)
screening and integrate the ACE screening tool into routine prenatal visits
for pregnant women; bridging a gap between efforts of the county health
department and OPC.
Methods Used: Two years ago, Jefferson County Public Health (JCPH)
implemented ACE screening for pregnant women based on research from
seminal studies from the Centers for Disease Control and Kaiser Permanete.
This research correlates early childhood trauma (i.e. higher ACE scores) to
higher rates of chronic disease, mental illness, and poor health status later in
life. Medical staff at OPC were interested in ACE screening, but had not
implemented it.
ACE screening was researched in the medical literature and the method
for ACE screening at JCPH was reviewed. A training session on ACE
screening was developed and provided to OPC medical staff. The initial target
population for screening was identified to be pregnant women. An ACE
screening template was developed for use in the clinic. A web application
(www.acesurvey.org) was developed for use on smart phones which providers can use as a prompt for ACE screening when outside the clinic
(e.g. home or hospital visits). The ACE screening tool was piloted in the
clinic by screening two prenatal patients.
Summary of Results: Medical staff are ready to implement ACE screening
at the initial prenatal visit for new prenatal patients or the next scheduled visit
for existing prenatal patients. Medical staff are able to access the ACE
screening tool in the clinic or through the use of a web application on a smart
phone when outside the clinic.
143
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Conclusions: ACE screening for pregnant women was implemented at
Olympic Primary Care, the main providers of prenatal care in the area. This
project builds on established methods in the literature and work that had been
done by county public health leaders; bridging a gap between the efforts of
JCPH and OPC. Pregnant women who have the highest ACE scores can be
identified and referred to appropriate mental health services to help stop
intergenerational transmission of trauma.
Western Student Medical Research Forum
Student Session III - Adolescent Medicine,
Pediatrics, and Neonatal
8:30 AM
Friday, January 28, 2011
198
WOMEN INFANTS AND CHILDREN (WIC) POLICY CHANGE:
EFFECTS ON FRUIT AND VEGETABLE CONSUMPTION
AMONG LOW-INCOME CHILDREN NEW MEXICO
Greig E1,2, Davis S2, Myers O2 1UNM SOM, Albuquerque, NM and
2
University of New Mexico, Albuquerque, NM.
Purpose of Study: There is growing recognition of the increased need for
consumption of currently suboptimal levels of fruits and vegetables by
children. Intake is known to protect against chronic disease but low-income
children are at risk for decreased intake. Since the 1960_s WIC has offered
supplemental nutrition packages. The IOM recently reviewed WIC_s original
packages and found the need for improvement. One recommendation was to
increase funding for fruit and vegetables. As of October 2009 under the new
packages, participants will receive monthly vouchers for purchase of fruits
and vegetables. We wanted to investigate if the WIC policy change allows
participants to increase fruit and vegetable consumption as measured by
servings/day and if participants meet USDA guidelines for fruit and vegetable
intake.
Methods Used: Our study is an ancillary study to the Child Health Initiative
for Lifelong Eating and Exercise project, a NIH funded study focused on
obesity prevention and intervention. The participants were children ages 3Y5
from the reference arm of the intervention study. To examine the effects of
policy change we added questions to the parent interview to assess participation in WIC both pre/post policy change. To assess consumption we used a
24-hr dietary recall and food frequency questionnaire. A binary indicator of
whether reported intake was Q USDA guidelines (X servings for vegetables,
Y servings for fruit) was calculated. Summary means and standard deviations
were calculated for intake variables and relative frequencies were used to
summarize the intake goal variables.
Summary of Results: We identified 130 eligible participants during the
pre-policy period and 88 for the post-policy. Pre-policy group reported mean
fruit servings; 2.14T1.8, and mean vegetable servings; 1.61T1.34. Post-policy
group reported mean fruit servings 2.19T1.02, and mean vegetable servings
1.71T1.57. Relative frequency of servings meeting USDA guidelines goal
for fruit intake pre-policy was 51% and post-policy change was 55% and
for vegetable intake 8% and 15% respectively.
Conclusions: The WIC policy change showed minimal increase in fruit and
vegetable consumption among low-income children. Children in the study
did not meet USDA guidelines for number of servings/day.
199
CHARACTERIZING THE TRIGGERS OF PEDIATRIC
ASTHMA IN A RURAL ENVIRONMENT
Mandell RY1, Karr C2 1University of Washington, Seattle, WA and
2
University of Washington, Seattle, WA.
Purpose of Study: Asthma is an important pediatric health concern in the
US, affecting approximately one in 12 children. In the primarily Latino agricultural community of Yakima Valley, Washington, pediatric asthma has
been established as a top health concern by surveys of the community and
health care workers. While asthma triggers have been well characterized in
urban settings, there has been little focus on rural pediatric asthma, particularly among immigrant farm working families. This study sought to characterize asthma in a cohort of 41 children who participate in the Yakima
Valley Farm Workers Clinic Asthma Project. We hypothesized that the role of
ambient environmental exposures and established indoor triggers in a rural
144
&
Volume 59, Number 1, January 2011
community might differ from observations in urban inner-city children with
asthma.
Methods Used: Bilingual interviewers conducted a questionnaire on residential, agricultural, and other ambient exposures. We conducted skin prick
testing for inhalant allergens, exhaled nitric oxide measurements, and spirometric measurements on all subjects.
Summary of Results: The most frequently reported triggers of asthma
included: respiratory infection (92.7%); exercise (80.5%); pollen (68.3%);
household dust (68.3%); and cold weather (68.3%). Among agricultural
factors, participants reported triggering due to cut grass (41.5%); crop sprays
(34.1%); grain dust (29.3%); livestock (26.8%); and hops fields (17.1%).
Participants reported occupational exposures, with 35.0% reporting an adult
in the household exposed to dusts, gases, fumes, chemicals, or strong odors,
and 7.3% reporting that the child is regularly exposed when accompanying a
parent to work. In total, 17.1% said that work exposures contribute to the
child_s asthma. Questions were asked to determine environmental exposures
near to the home: 39.0% reported close proximity (G 3 mile) to crop farms;
22.0% to farms raising animals; 41.5% to major roads with heavy traffic; and
46.3% to dusty roads. Skin prick testing of 22 allergens in the region demonstrated that 75.6% of subjects are atopic and 24.4% are non-atopic.
Conclusions: This study suggests that children in a rural, agricultural setting
may face unique environmental triggers of asthma. Intervention strategies
derived from urban-based studies may require modification to best serve this
population.
200
BICEPS BRACHII LONG HEAD OVERACTIVITY
ASSOCIATED WITH ELBOW FLEXION CONTRACTURE
IN BRACHIAL PLEXUS BIRTH PALSY
Sheffler L1, Lattanza L2, Sison-Williamson M2, James M2 1University of
California, Davis School of Medicine, Sacramento, CA and 2Shriners
Hospital for Children, Northern California, Sacramento, CA.
Purpose of Study: The etiology of elbow flexion contracture in children
with brachial plexus birth palsy (BPBP) has yet to be established. We hypothesized that the biceps brachii long head assists with shoulder stabilization
in children with BPBP and that overactivity of the long head during elbow
and shoulder activity is associated with an elbow flexion contracture.
Methods Used: Twenty-one subjects with BPBP-associated elbow flexion
contracture underwent testing with surface and fine wire electromyography
(EMG). Surface electrodes were placed on the muscle belly and fine wire
electrodes were inserted into the biceps brachii long and short heads, bilaterally. Subjects were asked to perform four upper extremity tasks: elbow
flexion/extension, hand to head, high reach and overhead ball throw. Mean
duration of muscle activity in the affected limb was compared to the contralateral unaffected limb, which was used as a control. Three-dimensional
motion analysis, surface dynamometry and validated function measures were
used to evaluate upper extremity kinematics, elbow flexor-extensor muscle
imbalance and function, respectively.
Summary of Results: The mean activity duration of the biceps brachii long
head was significantly higher in the affected limb, as compared to the unaffected limb during hand to head (p=0.02) and high reach tasks (p=0.03). No
statistically significant differences in mean activity duration were observed
for the biceps brachii short head between the affected and unaffected limbs.
Isometric strength of elbow flexion was not significantly higher than that of
elbow extension in the affected limb (p=0.11).
Conclusions: Overactivity of the long head of the biceps brachii is associated with and may contribute to the development of elbow flexion contracture in children with BPBP. Elbow flexion contracture is not associated
with an elbow flexor-extensor muscle imbalance, as previously hypothesized.
The negative impact of elbow flexion contracture on upper extremity function
warrants future research in the development of preventive and therapeutic
techniques to address elbow flexion contractures in children with BPBP.
201
INCREASING ADOLESCENT SPORTS PARTICIPATION TO
PREVENT TOBACCO USE IN BUFFALO, WYOMING
Conway DM University of Washington School of Medicine, Seattle, WA.
Purpose of Study: In Wyoming, 14.9% of 12 to 17 year-olds use tobacco,
ranking the state worst in the nation. In Buffalo, 19.7% of middle school and
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
37.1% of high school students report having tried cigarettes. 11.7% of high
school students smoked three or more days in the last month and 25% used
smokeless tobacco. This project aimed to increase kids_ interest in sports
participation, specifically basketball, as a means of promoting healthy habits
and preventing tobacco use.
Methods Used: Buffalo_s public health needs were informally assessed
through discussions with various agencies and community members. With
tobacco use a recurring concern, the Buffalo 2009 Youth Risk Behavior
Survey was obtained and a critical review of the literature performed. This
showed sports participation correlates with decreased tobacco use and sports
teams are prime vehicles for curricula promoting healthy lifestyles. The
project partnered with the local YMCA to utilize gym space, marketing
materials, and scholarships. Anti-tobacco curriculum ideas were solicited
from tobacco cessation leaders, respiratory therapists, and online resources.
A basketball camp itinerary was created with professional support.
Summary of Results: A three-day basketball camp with daily tobacco
education activities engaged 15 boys and girls ages 6 through 12. Tobacco
curriculum included peer led activities, hands on experiments, an interactive
talk with a juvenile drug probation officer, and a presentation by a teenage nicotine addict. After active participation, feedback from campers indicated they received the message being shared. All campers articulated their
pledge to be tobacco free on a camp poster. Parents reported campers came
home and shared their newfound knowledge and brochures. Six participants
reported this was their first sports camp and all asked when the next one
would be. These inquiries were suggestive of future interest in sports
participation.
Conclusions: Community anti-tobacco campaigns previously existed in
Buffalo. None of these, however, have focused on sports involvement as
a means of prevention. This project elucidated to community leaders the
viability of this format for transmitting the anti-tobacco message. Children
need an outlet for their energy and an introduction to sports supports a
lifetime of healthy habit development, camaraderie, and positive peer support
networks.
202
VIOLENCE IN THE COMMUNITY: A COMPREHENSIVE
CURRICULUM TO PRODUCE YOUTH PEER-ADVOCATES
FOR VICTIMS OF VIOLENCE
Hunziker A University of Washington, Seattle, WA.
Purpose of Study: Providers at the Rainier Park Medical Clinic identified
violence as a significant health risk in the community. The literature indicates that inter-city youth, aged 12Y15, will encounter violence personally
or secondarily in their lives. The curriculum to produce youth peer-advocates
in the Rainier Valley neighborhood of Seattle, Washington was developed
to increase these youths_ ability to recognize violence, to prevent it, and to
access community resources to treat it.
Methods Used: Youth members from the Rainier Valley Boys & Girls
Club participated in the 5-day, 10-hour course. The curriculum incorporated
information about violence toward self and others from the professional
literature. Education was discussion-based to help students recognize the
emotional and physical health implications of violence. Discussions led by
community leaders included definitions of violence, how to identify violence,
the health effects of violence on a personal and community level, how to
access evidence-based literature electronic databases, how to access community resources, how to prevent violence, how to communicate with victims
of violence and how to direct them to the appropriate resources for intervention and treatment.
Summary of Results: Four of the five youth in the program demonstrated
their understanding of violence, how to recognize it_s effects, how to use
personal coping skills, how they might intervene, and how to access resources. These students earned Proficiency in Peer-Advocacy certificates.
All of the youth received service learning hours to apply to their high
school graduation requirements and gift cards donated by the community
partners. Copies of the curriculum were requested by community partners
in order to repeat the peer-advocacy training.
Conclusions: This age group is interested and can learn in an interactive,
fast-paced community based program. The incentive of gaining service
hours and gift cards was important for registering participants but did not
appear to impact student involvement. Partnering with the community
was key to success and offered a forum for collaboration around youth
violence.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
203
INCREASING ACCESS TO ADOLESCENT REPRODUCTIVE
HEALTH SERVICES ON PRINCE OF WALES ISLAND,
ALASKA
Martinez CL 1University of Washington School of Medicine, Seattle, WA
and 2Alicia Roberts Medical Center, Klawock, AK.
Purpose of Study: Adolescents on Prince of Wales Island (POW) face
multiple barriers to accessing reproductive health services. Barriers include
great distances between island communities, a perceived lack of confidentiality and absence of comprehensive sexual education in schools. Native
teens are more likely to engage in sexual activity and less likely to use
contraceptive methods than non-Native teens. As a result, Natives experience
a disproportionately high rate of STDs. Alaska Natives make up 18% of the
state_s population, yet they account for 46% of Chlamydia cases. This intervention aims to reduce STD rates through education and increased access
to preventive services.
Methods Used: Interviews were conducted with health care providers, tribal
leaders and teens in order to identify barriers to adolescent services. Confidentiality concerns, geographic barriers and lack of education were identified
as factors adversely affecting adolescent health outcomes. A literature review
was conducted to determine sexual health risks facing Alaska Natives and
culturally-specific prevention strategies. Teen clinics were held in the Native
villages of Klawock and Hydaburg, offering education, contraceptive services and STD screening. A POW Youth Resource Guide was developed to
increase awareness and utilization of existing non-profit and tribal programs.
Summary of Results: Teen clinics were held in the Tlingit village of
Klawock and the Haida village of Hydaburg, increasing access to reproductive health services. Teen Resource Guides were distributed at various
sites around the island.
Conclusions: Health care providers and community leaders of POW recognize the need for comprehensive sexual education and preventive services.
However, some community members still remain resistant to such efforts.
Residents may respond more positively if interventions come from within
their village, rather than from outside health care providers. It is recommended that local area health clinics work with tribal leaders and community
members to identify possible educators within the village. It is also suggested
that schools incorporate comprehensive sexual education into their curriculum, rather than solely abstinence-based programs in order to promote open
dialogue about sexual activity and preventive practices.
204
ALCOHOL ABUSE PREVENTION AMONG ADOLESCENTS
IN COTTONWOOD, ID
Parden K University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Alcohol is the most commonly used drug among rural
adolescents in the United States. In Cottonwood, a small farming community,
30% of the population is under 18 years old. Early initiation of alcohol
consumption greatly increases the probability of alcohol-related problems
later in life, thus the alcohol abuse prevention program was created to target
adolescents. The goal was to provide an interactive discussion session with
students to encourage the development of critical thinking and social coping
skills as well as provide education about the physical effects and dangers of
alcohol abuse.
Methods Used: The need for alcohol abuse prevention was determined
after questioning community members and St. Mary_s Clinic staff. A review
of the literature revealed effective methods of adolescent alcohol and substance abuse education, including how to design an interactive session that
encouraged the development of refusal skills. Sports physical nights held in
Cottonwood, Nezperce, and Kamiah by the St. Mary_s Clinic were utilized
to achieve contact with a target population of 6Y12th graders. Each adolescent participated in small-group or one-on-one discussions about alcohol
abuse. Discussions included tactics for handling peer pressure, the adolescent_s own experiences with alcohol, and education about the physical
effects of alcohol consumption on health and athletic ability. Fatal vision
goggles were obtained from the Moscow Police Department to help create
an interactive environment. A brochure was developed to reinforce the discussion and answer additional questions.
Summary of Results: Approximately 138 adolescents participated in
the alcohol abuse prevention discussions at the sports physical nights in
Cottonwood, Nezperce, and Kamiah. Many of the students had not previously
145
Western Regional Meeting Abstracts
Journal of Investigative Medicine
considered the session material and subsequently benefited both from discussing peer pressure situations and from learning more about the physical
effects of alcohol.
Conclusions: An interactive discussion was a helpful format for alcohol
abuse prevention education among adolescents. It encouraged the students to
consider the physical effects of alcohol consumption and to practice refusal
skills. The fatal vision goggles were an effective method of initiating discussion. The prevention program could be retained and even expanded upon
for future education.
205
LIVER EXPRESSION OF IGF-1 IS NOT DECREASED IN
PRETERM LAMBS VENTILATED FOR 3 DAYS AND WEANED
FROM VENTILATION
Block C, Dahl M, Von Der Ahe N, McCoy MJ, Wang Z, Dong L, McKnight
R, Null D, Yoder BA, Lane RH, Albertine K University of Utah, Salt Lake
City, UT.
Purpose of Study: Growth is modulated by insulin-like growth factor-1
(IGF-1). Serum levels of IGF-1 are low in preterm infants with neonatal CLD.
Hepatic expression of IGF-1 determines serum levels. We showed that 21d of
mechanical ventilation (MV) resulted in poor growth and diminished expression of IGF-1 in the liver of preterm lambs (J Invest Med 58:105, 2010).
Liver expression of IGF-1 was also diminished at 3d of MV, when alveolar
simplification was evident in the lung of the same preterm lambs. Whether
liver IGF-1 expression is affected by 3d of MV followed by weaning from
ventilation is not known.We hypothesized that mechanical ventilation for 3d
will lead to long-term decrease in IGF-1 and its upstream signaling molecules
Janis kinase 2 (JAK2) and signal transducers and activators of transcription
5b (STAT5b).
Methods Used: Preterm lambs (È128d gestation; term È150d; n=4), treated
with antenatal steroids and postnatal surfactant, were managed by MV for 3d,
weaned using high-frequency nasal ventilation for 3d, and then weaned from
ventilation and recovered for 10 wk (equivalent to È2 yr postnatal life
in humans). Liver tissue was analyzed by quantitative real time RT-PCR
(normalized GAPDH mRNA expression) and immunoblot (normalized
MemCode). Control lambs were delivered at term and lived 8 wk (n=4).
Summary of Results: At the end of the 11wk study period, the weight of
preterm weaned lambs and control lambs was the same (21T6 vs 22T6 Kg,
respectively). Liver tissue from weaned preterm lambs and control lambs had
the same expression of IGF-1 mRNA (0.7T0.1 vs 0.6T0.1, respectively) and
STAT5b mRNA (0.3T0.1 vs 0.3T0.01, respectively). Likewise, weaned preterm lambs and control lambs had the same relative abundance of STAT5b
protein (10T8 vs 10T3, respectively) and JAK2 protein (77T6 vs 66T11,
respectively).
Conclusions: Preterm birth followed by 3d of MV and subsequent weaning
from ventilation support did not cause sustained growth delay or liver expression of IGF-1 and its upstream signaling molecules. Despite this sparing
effect on the liver of preterm weaned preterm lambs, the lung of the same
preterm weaned lambs had sustained delay in alveolar formation that is
characteristic of neonatal CLD. (J Invest Med 57:223, 2010). (HL62875,
HL56401, HD41075).
206
IL-17 PRODUCTION BY CORD BLOOD MONONUCLEAR
CELLS IS DIMINISHED COMPARED TO ADULTS
Caron JE1,2, La Pine TR1,3, Augustine NH1,2, Martins TB2, Hill HR1,2,3
1
University of Utah, Salt Lake City, UT; 2ARUP Institute for Clinical and
Experimental Pathology, Salt Lake City, UT and 3University of Utah, Salt
Lake City, UT.
Purpose of Study: Human neonates are uniquely susceptible to severe and
overwhelming bacterial and fungal infections. Defective neonatal polymorphonuclear leukocyte (PMN) activation and movement contribute to this
increased susceptibility to infection. T helper 1 (Th1) lymphocytes produce
interleukin 17 (IL-17), which is thought to act on fibroblasts and endothelial
cells to recruit neutrophils into local areas of microbial invasion. Previously,
we have demonstrated that cord blood mixed mononuclear cells have defective production of the Th1 cytokines IL-18 and IFN-F.
Here we examined cord blood mixed mononuclear cell production of
IL-17, a cytokine critical in the host response to bacterial and fungal infections, and compared the results with that of mononuclear cells from adults.
146
&
Volume 59, Number 1, January 2011
Methods Used: Whole blood was collected from healthy adults and umbilical cord blood from healthy term deliveries. Mixed mononuclear cells
(MMCs) were isolated on Ficoll-Paque and stimulated with phytohemagglutinin (PHA), a potent mitogen, and then incubated for 24 hours in tissue
culture medium. IL-17 production was measured using a new, in-house developed assay utilizing Luminex multianalyte technology.
Summary of Results: Mononuclear cells from newborn infants
(0.10 pg/mL +/j 0.1) produced significantly less IL17 than mononuclear
cells from adults (309 +/j 2.94 SEM pg/mL; pG0.02).
Conclusions: IL17 has a profound effect on the immune response to bacterial and fungal infections. For instance, IL17 production is profoundly
deficient in autosomal dominant Hyper IgE or Job syndrome in which
patients suffer repeated bacterial and candida infections. The present report is
the first of a deficiency in IL17 production by neonatal mononuclear cells,
which likely contributes significantly to the increased susceptibility of human
infants to microbial infections.
207
GLUCOSE TRANSPORTER mRNA EXPRESSION IN
PLACENTAS OF GROWTH-RESTRICTED FETUSES
Kim JE1, Cho J1, Shin B2, Devaskar S2, Janzen C1 1David Geffen School of
Medicine at UCLA, Los Angeles, CA and 2David Geffen School of Medicine
at UCLA, Los Angeles, CA.
Purpose of Study: Intrauterine growth restriction (IUGR) is the inability of
a fetus to achieve proper growth. IUGR is associated with increased perinatal
morbidity and mortality. Glucose is an essential nutrient for fetal growth and
is transported from mother-to-fetus across the placenta. Various facilitated
glucose transporters (GLUTs) are present in the placenta, including the
isoforms GLUT1, GLUT3, and GLUT4. Recent mouse studies demonstrated
that homozygous GLUT3 null mutations resulted in early pregnancy loss,
while GLUT3 heterozygotes exhibited fetal growth restriction. The objective
of this study was to determine if mRNA expression of GLUT1, GLUT3, and
GLUT4 in human placenta changes with IUGR.
Methods Used: In this prospective study, human placentas were collected at
time of delivery. GLUT1, GLUT3, and GLUT4 mRNA expression in the
maternal and fetal regions of each placenta was determined by quantitative
real-time PCR. Results were quantified by the comparative CT method and
statistical analysis was conducted using the Student_s t-test.
Summary of Results: Placentas were collected from control groups (n=3,
birth weight percentile 65.3T28.0) and IUGR groups(n=7, birth weight percentile 4.7T3.1). qRT-PCR results showed no significant difference in mRNA
expression levels of GLUT1, GLUT3, or GLUT4.
Conclusions: In this study, significant differences were not found in the
mRNA expression of glucose transporters in human placentas affected by
IUGR. However, there appeared to be a trend towards increased GLUT1
and GLUT3 expression with IUGR. Additional studies with increased
sample sizes are necessary to determine if the observed differences are
significant.
208
PATIENT/FAMILY CHARACTERISTICS IN AMERICAN
INDIAN CHILD AND ADOLESCENT QUALITY OF
CARE RESEARCH
Podlogar M1, Novins DK2 1University of Colorado Health Sciences Center,
Aurora, CO and 2University of Colorado Health Sciences Center, Aurora, CO.
Purpose of Study: Across the country, both rural and urban American
Indian (AI) children and adolescents are at a higher risk than other U.S. ethnic
groups for developing mental health problems such as depression, substance
abuse, domestic violence, and suicide, and have a disparately high need for
mental health services. New tribal healthcare service systems, as well as state
and federal agencies (such as Medicaid and the IHS), could highly benefit
from a systematic assessment of the quality of the mental health care they
provide to AIs. Perhaps the best known framework is that proposed by
Donabedian, which organizes the assessment of care into an assessment of its
structure, process, and outcome. Additionally, health policy makers, providers, and consumers have recognized that culturally competent care addresses
and eliminates racial/ethnic disparities in health care. We propose to introduce a fourth domain, Bpatient/family characteristics,[ to Donabedian_s
framework, and to investigate how this domain relates to the other domains of
assessment, as well as to cultural appropriateness of care.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Methods Used: Data extracted from medical records of three research sites
(with a subsample verified by an independent rater as a quality control
mechanism) and interviews with administrators, clinicians, parents, and
youth will be analyzed with the aid of qualitative analysis programs such as
NVivo, looking for clinician, patient, and family perspectives of treatment in
regards to the four examined domains of quality of care.
Summary of Results: Collaboration with representatives from the partner
sites and preliminary analysis of data extracted from medical records has
defined patient/family characteristics to include sociodemographics, family
structure, specific behavioral health difficulties, attitudes towards treatment,
and barriers or support to care. Further research will analyze the relationship
of these characteristics to structure, process, and outcome.
Conclusions: We hypothesize that the relationship between patient/family
characteristics and structure, process, and outcome of mental health services
will provide a useful model for subsequent studies in more culturally sensitive
AI quality of care research.
Adolescent Medicine and General Pediatrics
Concurrent Session
1:00 PM
Friday, January 28, 2011
209
BICYCLE AND ATV HELMET SAFETY IN MCCALL, IDAHO
Hallett H University of Washington, Seattle, WA.
Purpose of Study: McCall is a rural town of about 2,600 people nestled
against the Payette National Forest. McCall_s picturesque scenery and access
to the outdoors has made it a tourist destination. Because of its proximity to
skiing, hiking, biking, and ATV riding, many McCall residents, as well as
their children, are outdoor enthusiasts. While helmet use decreases head
injury risk in bicycle and ATV crashes by 88% and 64%, respectively, none
of McCall_s sporting good stores currently offer helmet safety courses.
The purpose of this project was to create an interactive ATV and bicycle
helmet program for 6th grade students at Payette Lakes Medical School
that addressed barriers to helmet use, basic brain anatomy and deficits seen
with brain damage, and proper ATV and bicycle helmet fit.
Methods Used: A literature review was performed to determine prevalence
of ATV and bicycle injuries, helmet use, and possible educational interventions. After consulting with the chairman of the school board and the Payette
Lakes Middle School health teacher, a bicycle and ATV helmet safety program was developed and presented to 6th grade students. Students were
polled about their bike and ATV helmet practices. An activity was performed
to simulate falling off a bike or being thrown from an ATV using hard boiled
eggs, to represent the brain, wrapped with materials to represent the meninges
and skull. Half the eggs were equipped with plastic helmets, and students
compared egg damage with and without helmet use. Students were guided
through the steps of properly fitting ATV and bicycle helmets and then asked
to fix incorrectly placed helmets.
Summary of Results: Sixty-five 6th grade students participated in the
bicycle and ATV helmet safety program. Students gave feedback on what
they learned with Bbike helmets provide 88% protection from head injury[
as the most common response. A flier on proper ATV and bicycle helmet
fit was sent home with students to help them educate their parents. The
curriculum was given to the 6th grade teachers and the Payette Lakes Medical
Clinic to continue the program next year.
Conclusions: Because helmet use is low in middle school kids, an intervention to effect change had to be interactive and meaningful. By connecting
brain function with brain damage in ATV and bicycle crashes, the students
gained a better understanding of the purpose of helmets.
210
MATERNAL RISKS, COMMUNITY VIOLENCE EXPOSURE
AND SELF-REPORTED ASTHMA AMONG CHILDREN IN
FOSTER CARE
Hellyer J, Culhane SE, Garrido EF, Petrenko CL, Taussig HN University of
Colorado Denver, Aurora, CO.
Purpose of Study: The occurrence of pediatric asthma has been associated
with exposure to chronic stress. The goal of this study was to examine
whether stress and asthma were associated in a high risk sample. The study
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
examined the relationship between maternal and community risk factors and
asthma in a sample of maltreated children placed in foster care.
Methods Used: Interviews were conducted with 355 maltreated children
(9Y11 years old) who had been court-ordered into foster care within the
past year. Measures included youth self-report of asthma and community
violence exposure and an index of maternal risk (criminal history, incarceration, alcohol and controlled substance use, domestic violence perpetration, domestic violence victim, mental illness, inadequate housing, history of
foster care placement and history of maltreatment) based on data abstracted
from child welfare records.
Summary of Results: After controlling for age, gender and ethnicity,
maternal risk significantly predicted the presence of asthma (OR = 1.26, 95%
CI = 1.04Y1.52, p=0.02) over and above the effects of community violence
exposure, but community violence exposure did not predict asthma over and
above the effects of maternal risk.
Conclusions: Maternal risk factors were significantly associated with the
presence of asthma in a maltreated, foster care population. This connection
has important implications for clinicians who oversee the care of maltreated
children as it may be an indication of which children in this already high-risk
group have a greater propensity for physical health issues.
211
INTRATUBULAR GERM CELL NEOPLASIA IN THE
PEDIATRIC POPULATION: A CASE REPORT
DeGirolamo K1,2, Masterson J1,2 1University of British Columbia,
Vancouver, BC, Canada and 2BC Children’s Hospital, Vancouver, BC,
Canada.
Case Report: Testicular cancer, specifically germ cell tumors, are the most
common malignancy in young men and are curable in up to 95% of cases, but
the incidence is on the rise. (1) The key event in the transformation to
malignancy is hypothesized to occur in early adulthood and perhaps the
carcinogenic pathway for germ cell tumors begins in fetal life due to maternal
estrogen or exposure to environmental estrogens. (1) Intratubular Germ Cell
Neoplasia (ITGCN), previously known as Carcinoma in Situ of the testis, is a
premalignant condition that can progress to testicular cancer and is associated
with testicular germ cell tumors. CIS was first discovered after Skakkebaek
biopsied testicles of infertile men who later developed testicular germ cell
tumors. (2) These cells are thought to be more susceptible to mutations from
changing hormone levels and thus germ cell tumors typically occur after
puberty or in patients with sexual development disorders, such as feminization of the testis. (1) This is a rare occurrence in prepubertal children and it
would be ideal to know the prognostic implications. We present a case report
of a 4-month-old male with undifferentiated ITGCN, and a co-morbid mature
teratoma. To the best of our knowledge this is the first case documented of
this kind and demonstrates how little is known about this condition and how
this specific patient was managed surgically and clinically.
1. Emerson R and Ulbright T. Intratubular Germ Cell Neoplasia of the
Testis and its Associated Cancers: The Use of Novel Biomarkers.
Pathology June 2010;42(4):344Y55.
2. Skakkebaek, NM. Possible Carcinoma-In-Situ of Testis. Lancet
1972;2:516Y7.
212
LEARN PEDIATRICS: CREATING A WEB-BASED
MULTIMEDIA RESOURCE FOR MEDICAL STUDENTS
LEARNING PEDIATRIC CLINICAL SKILLS
Jekyll A, Statham E, Bishop J, Macnab A, Collet J, Hurley RM University of
British Columbia, Vancouver, BC, Canada.
Purpose of Study: Few non-profit online resources currently exist for
teaching medical students pediatric clinical skills. Computer-based education is effective at teaching physical examination in addition to being costeffective. The objective is to create a free, online resource containing written
material, videos and other multimedia formats to assist medical students
with learning pediatric clinical skills and approaches to common pediatric
presentations. The long-term goal is to establish and maintain an open domain,
non-profit website that will enhance the learning of medical students nationally
and internationally.
147
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Methods Used: The website, www.learnpediatrics.com, consists of ten
Purpose of Study: Patterns of growth amongst a nation_s children most
pediatric modules: general, respiratory, cardiology, gastroenterology, newborn, genitourinary, neurology, musculoskeletal, hematology/oncology, and
endocrinology. Seven of these modules will contain physical exam teaching
videos. Medical students write content on a voluntary basis and receive
credit for their contributions. Pediatric residents then edit the articles for
content and clarity. Video scripts are written by pediatric residents, in
consultation with attending staff. All actors involved in the videos are
volunteers. The University of British Columbia (UBC) Media Production
records the narrations, films the clinical teaching videos and performs final
edits with a pediatric resident or staff physician. Final written and multimedia materials are posted to the site by students employed by the faculty
in work-study arrangements.
Summary of Results: Learn Pediatrics has been successfully transferred to
the UBC domain. Completed videos including respiratory, newborn, cardiology and neurology have been posted. The scripts are written for both
gastroenterology and genitourinary. All videos and scripts have been edited
by content experts. Seventy content topics out of 125 have been written by
students and edited by pediatric residents. There is ongoing involvement to
complete writing of the content topics.
Conclusions: Learn Pediatrics is a free, web-based resource for medical
students learning pediatric clinical skills and is envisioned to become a
valuable resource for medical students not only at UBC, but worldwide.
Future directions include self-assessment tools and evaluation of the website.
accurately reflect the nutritional and health status of its population and predict
the health of future generations.
Methods Used: Children enrolled in BBrighter Smiles[, a collaborative
health-promotion program, were evaluated for parameters of growth. Height,
weight, BMI and health data via questionnaire were obtained for children at a
primary school in rural Uganda to compare their status to the WHO Child
Growth Standard (2006). The WHO Standard uses multiethnic sampling
(populations from 6 countries) to capture the genetic variability among
continents and generate a single international growth standard. This standard
is intended to underscore that differences in nurture, rather than nature, determine disparities in physical growth, and provide a basis for appropriate
healthcare policy and intervention.
Summary of Results: 233 children (105 males, 128 females), aged between 5Y13 years, were evaluated. Amongst the girls 91% were below the
50th centile for height and 90% for weight. Similarly, amongst the boys 89%
and 88% were below the 50th centile for height and weight respectively. A
high proportion of these children come from single parent or child-headed
families (this community was the epicentre for the AIDS epidemic) and
poverty, borderline nutrition, malaria, and parasitic infections are prevalent.
Prior research here has also documented that cultural and religious beliefs
lead to restricted intake of essential foods, and both low food quality and
infectious diseases commonly exacerbate malnutrition.
Conclusions: The finding that such a high percentage of our cohort_s
children had height and weight below the 50th centile most probably
reflects the impact of local social circumstances, sub-optimal nutrition and
infectious disease, and indicates the value and potential for the WHO
standards to identify where children_s growth patterns would benefit from
interventions to promote appropriate nurture, and improve health and
growth potential.
213
SMOKING PREVENTION IN ADOLESCENT GIRLS
Ball AJ University of Washington School of Medicine, Seattle, WA.
Purpose of Study: MontanaVthe land of the cowboyVhas high rates of
tobacco use. Marias Medical Center in Shelby, MontanaVa town of approximately 3,500 residentsVsees adolescents who feel cultural pressures to
smoke cigarettes almost daily. Marlboro_s 2007 BCamel No. 9[ advertising
campaign suggests that adolescent girls are actively being targeted for cigarette advertising. The purpose of this project was to promote evidencebased, age and gender appropriate smoking prevention strategies.
Methods Used: Discussions with family doctors, nurses, Toole County
public health department employees, and observing clinic visits were
methods used to identify non-smoking adolescents as the community project
focus. A grassroots approach was taken to reach local youth, including the
submission of a letter to the local paper and a day spent at the Shelby youth
swim meet encouraging swimmers to make a tobacco-free pledge. To develop
local awareness of the harms of smoking, efforts were combined with the
Toole Country public health department at their open house. Youth-friendly
Bprops[ were used to facilitate meaningful discussions with pre-teens. A
literature review was performed for validation of the recommendation of
nosmokingroom.org, an educational Bgame,[ and guidance in choosing
methods and content of communication with the target group.
Summary of Results: A letter to readers of the Shelby Promoter, the local
weekly newspaper, encouraged local adolescents seeking tobacco information to take advantage of local physicians and internet resources such as
nosmokingroom.org, an anti-tobacco, interactive website for girls ages 8Y12
designed by pediatricians. At a youth swim meet, over thirty adolescents
from Shelby and surrounding towns each pledged to remain tobacco-free by
signing a shirt. T-shirts, frisbees, and water bottles featuring anti-smoking
messages were given to swimmers. Further tobacco information was provided
to residents at the health department_s open house.
Conclusions: New evidence-based age and gender appropriate strategies are
needed to prevent smoking in adolescents. Interactive methods such as collecting pledge signatures and game-like educational tools online add to the
tools of smoking prevention. However, more evidence is needed to support
educational gaming as an effective method of smoking prevention in adolescents before the strategy can be applied more globally.
214
BRIGHTER SMILES: A COMPARISON OF GROWTH IN
RURAL UGANDAN CHILDREN TO WHO STANDARDS
Zhang M1, Ashaba B2, Berg T1, Choi W1, Kasangaki A2, Musinguzi N2,
Meredith C1, Nambatya B2, Nyairo S2, Cannon W1, Macnab A1 1University
of British Columbia, Vancouver, BC, Canada and 2Makerere University,
Kampala, Uganda.
148
215
DISPARITY IN HEALTH LITERACY BETWEEN ENGLISH
SPEAKING AND SPANISH SPEAKING PARENTS OF
HOSPITALIZED CHILDREN
McDermott J1,2, Joseph M1,2, Poynter R1 1Children’s Hospital Central
California, Madera, CA and 2UCSF Fresno Medical Education Program,
Fresno, CA.
Purpose of Study: Health literacy is defined as Bthe degree to which
individuals have the capacity to obtain, process, and understand basic health
information and services needed to make appropriate health decisions[,
according to Healthy People 2010. Low socioeconomic status, lower education levels, and English as a second language are factors that correlate
with lower health literacy in adults. However, few have studied disparities
in parental health literacy when comparing groups by primary language,
an important factor that could adversely affect child health outcomes. We
hypothesize a significant difference exists between health literacy rates of
English speaking and Spanish speaking parents in a hospital setting.
Methods Used: The study is being conducted in a tertiary children_s hospital in the Central Valley of California. We aim to study 200 parents, 100
English speaking and 100 Spanish speaking. Parents of patients admitted on
the inpatient pediatric medical services are being recruited. Health literacy
scores are assessed using the BNew Vital Sign[ tool, a 6 item survey validated
in English and Spanish. The same person is administering the surveys to all
subjects; a certified Spanish translator is used to administer the survey to
Spanish speaking parents. All subjects also complete a brief questionnaire to
obtain demographics. Statistical analysis is done using Chi-Square with
Fisher_s Exact Test.
Summary of Results: Results of 43 completed surveys from 21 (49%)
Spanish-speaking parents and 22 (51%) English speaking parents have
been analyzed. Analysis of our initial data shows the possibility of limited
literacy in 11 (52%) in Spanish speaking parents compared to 2 (9%) in
English speaking parents (p=0.003). Data collection is ongoing and will
be completed within two months. Final results will be discussed at the
meeting.
Conclusions: Preliminary analysis reveals significantly lower health literacy
in Spanish speaking compared to English speaking parents. We expect the
same trend to continue in the remaining study population. With the information gained from this study, we plan to educate our medical staff and
support services in identifying parents at risk for low health literacy and
providing linguistically appropriate resources.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Conclusions: Although an increased risk of acute injury in overweight and
216
KEY ELEMENTS FOR SUCCESSFUL GLOBAL HEALTH
EDUCATION ELECTIVES: LESSONS FROM BBRIGHTER
SMILES AFRICA[
Zhang M1, Berg T1, Ashaba B2, Berg M1, Choi W1, Meredith C1, Musinguzi
N2, Nambatya J2, Nyairo S2, Cannon W1, Kasangaki A2, Macnab A1
1
University of British Columbia, Vancouver, BC, Canada and 2Makerere
University, Kampala, Uganda.
Purpose of Study: Global health electives have the potential to foster
opportunities for service learning and create a platform to educate students
on the key determinants of health, and cultural competencies and logistics
necessary for successful program delivery. To be effective and relevant to the
host country such electives require structure.
Methods Used: We describe the key elements (identified via annual evaluation) central to success of a five year global-health partnership (Brighter
Smiles) between the University of British Columbia and Makerere University
in Uganda.
Summary of Results: SERVICE DELIVERY - School-based health promotion is an effective model endorsed by WHO. Giving interactive workshops provides knowledge and skills for healthy practices and promotes
change in school health culture. This informs and empowers the next generation on a broad range of health and social issues. COMMUNITY-BASED
LEARNING - Clinical opportunities and collaborative research/evaluation
provide new knowledge. Students learn the priorities and realities of medical care, prevention and social and cultural practices, establish the effectiveness of service delivery programs, and provide feedback and future
direction to participating individuals and communities. COLLABORATION Partnership of teams from different universities with local community leaders
during project delivery and workshops generates invaluable dialogue and
learning opportunities. Insights into realities and opportunities are gained,
and essential skills learned re team-work, communication, and cultural sensitivity. MENTORSHIP - Passion and experience shared by local and visiting
faculty is infectious and invaluable. This is an essential component to ensure
safe, culturally-relevant and sustainable student experiences.
Conclusions: Global health electives based on the FBrighter Smiles_ template should prove to be relevant, and offer effective and appropriate learning
for university students.
217
BODY MASS INDEX AND ACUTE INJURY RISK IN
CHILDREN AGE 5-17 YEARS
McRae L1,2, Campbell J3, Doan Q2, Kissoon N2, AlQahtani A2 1University
of British Columbia, Vancouver, BC, Canada; 2BCCH, Vancouver, BC,
Canada and 3UBC, Vancouver, BC, Canada.
Purpose of Study: Obesity is an epidemic that affects many Canadian children. Between 1981 and 2001 the prevalence of overweight and obese children
in Canada increased by 200Y300%(1). In 2004, 18.1% of Canadian children
age 2Y17 were overweight and 8.2% were obese(1). Obese children have
prolonged recovery times and increased morbidity and mortality following
an acute injury(2). The relationship between obesity and risk of acute injury
has not yet been established but is essential for proper weight loss counseling
of youth. Our study investigates the risk of acute injury associated with increasing body mass index (BMI) in children.
Methods Used: This is a case based, case control study with patients_ age 5
to 17 years who presented to BC Children_s Hospital Emergency Department
from July 2009 to December 2010. Cases were defined as children who
presented for assessment of an injury. Controls were children with any other
complaint who had not sustained an injury in the previous 12 months. Height
and weight were measured and BMI categories (underweight, normal, overweight and obese) were assigned according to the CDC classification per age
and sex. Descriptive statistics were used to report distribution of participants_
characteristics (BMI, sex, age, physical activity, socioeconomic status).
Multivariate logistical regressions estimated the odds of injury occurrence by
BMI category adjusted for confounders.
Summary of Results: To date we have screened 500 patients and 394 were
enrolled. Although the proportion of male subjects (45%) and BMI category
distribution (22Y23% overweight and obese combined) was similar among
cases and controls, the distribution for the other variables were considerably
distinctive. The control population is younger, less active and less affluent.
Results of logistic regression analyses are pending.
* 2011 The American Federation for Medical Research
obese children cannot be concluded from our descriptive analyses, a full
analysis is pending. 1. Starky S. The obesity epidemic in Canada. {http://
www2.parl.gc.ca/Content/LOP/ResearchPublications/prb0511-e.htm#
introductiontxt} (Accessed Sept 23, 2010). 2. Pomerantz W, Timm N,
Gittelman M. Injury patterns in obese versus nonobese children presenting
to a pediatric emergency department. Pediatrics 2010;125(4):681Y85.
218
CONCUSSION EVALUATION METHODS AMONG
WASHINGTON STATE HIGH SCHOOL FOOTBALL COACHES
AND ATHLETIC TRAINERS
Murphy A1, Kaufman MS2,4, Coppel DB3, Molton I2, Herring SA2,3,4
1
University of Washington School of Medicine, Seattle, WA; 2University of
Washington Medical Center, Seattle, WA; 3University of Washington
Medical Center, Seattle, WA and 4University of Washington Medical Center,
Seattle, WA.
Purpose of Study: To determine concussion assessment methods among
Washington State high school football coaches and athletic trainers.
Methods Used: A Catalyst WebQ survey link was emailed to varsity head
football coaches and athletic trainers from 106/298 Washington State school
districts. The survey was voluntary and anonymous. Survey questions inquired about participants_ concussion management training, familiarity with
the Zackery Lystedt Law, and standardized tools used for concussion assessment both on-the-field and for follow-up.
Summary of Results: Twenty-eight of 59 respondents (47%) used the
Standardized Concussion Assessment Tool 2 (SCAT2) for on-the-field assessment; urban respondents were statistically significantly more likely to use
SCAT2 (pG0.05). Neurocognitive testing (NCT) was used by 19/59 (32%)
respondents. This was statistically significantly used more commonly by
those in urban districts (pG0.001). A neuropsychologist interprets test results
for 11/19 (57.9%). There is no statistically significant correlation between
years of experience and use of the SCAT2, but those with more than 10 years
experience were less likely to use NCT (pG0.01). All participants reported
being familiar with the Zackery Lystedt Law, but only 44.1% reported that the
law has changed their concussion management.
Conclusions: There are statistically significant differences between SCAT2
and NCT use in urban and rural districts, as well as NCT usage amongst
respondents with varying years of experience. Further understanding and
identification of barriers limiting identification and management of concussions in high school athletes is crucial to ensure proper return to play and to
prevent serious permanent injury. Additional education is necessary to ensure
that athletic trainers and coaches are aware of current recommendations by
the International Concussion Conference for the evaluation and management
of concussions.
219
ATHLETIC PARTICIPATION AND SUBSTANCE USE IN
UNIVERSITY STUDENTS
Hsii A1, Carlson J1, Peebles R1,2 1LPCH, Palo Alto, CA and 2CHOP,
Philadelphia, PA.
Purpose of Study: Athletic participation may protect against substance use
in high school students; however, the data is conflicting at the collegiate level.
We examined the associations between athletic participation and substance
use in university students, and the substance use patterns in team-based and
individual-based competitive sports.
Methods Used: US university students (18Y26y;661M,973F) were recruited
through a social networking website to complete an online questionnaire on
athletics and health. Competitive athletes (CA) participated in a competitive
sport within the past year; recreational athletes (RA) exercised independently
and not as part of a team; sedentary students (SS) did neither. CA were further divided into team-based and individual-based sports. Prevalence of alcohol use, binge drinking, cigarette smoking, other tobacco use, marijuana use,
and other drug use were compared between groups using chi-squared analyses.
Summary of Results: There were 969 CA (429M,540F;402 team-based,
565 individual-based), 502 RA (157M,344F), and 166 SS (75M,89F). Alcohol was the dominant substance used, with CA and RA drinking and
binge drinking more than SS (alcohol: 86.8% vs 88.9% vs 76.3%, pe0.001;
binge drinking: 55.5% vs 54.9% vs 39.1%, pe0.001), with the greatest binge
drinking among female RA (53.6%) and male CA (63.0%). There were no
149
Western Regional Meeting Abstracts
Journal of Investigative Medicine
differences in cigarette use. Overall, other tobacco use was greatest in CA
and least in RA (7.0% vs 3.3% vs 6.4%, pe0.05) with higher use in males
(14.4% vs 10.1% vs 13.0%) than females (1.1% vs 0.3% vs 1.2%). Marijuana
consumption was least in CA with an increased trend in RA (24.5% vs 30.4%
vs 25.6%, p=0.06) with significance found in females (22.3% vs 30.1% vs
23.3%, pe0.05) but not in males. There were no differences in other drug
use. Overall substance use was greater in team-based than individual-based
athletes, with significant differences in binge drinking (64.3% vs 43.5%,
pe0.001), cigarette smoking (21.6% vs 12.5%, pe0.001), other tobacco use
(9.6% vs 3.3%, pe0.001), and marijuana use (30.6% vs 16.1%, pe0.001),
but not in alcohol use or other drug use.
Conclusions: Physical activity participation in these university students
did not protect against substance use. Collegiate substance use and awareness programs should consider focusing on athletic participants and should
consider gender differences as well.
220
IDENTIFICATION AND MANAGEMENT OF OVERWEIGHT
AND OBESE PEDIATRIC PATIENTS HOSPITALIZED
IN AN ACADEMIC HOSPITAL
King MA, Nkoy F, Halbern S, Maloney C University of Utah, Salt Lake
City, UT.
Purpose of Study: Obesity is the most common pediatric chronic illness
in the United States. Early identification and adequate management are
crucial steps in reducing related long-term complications and costs.
Studies have reported that outpatient provider compliance with identification of overweight/obesity is suboptimal. Hospitalization may represent an additional opportunity to identify, assess, and make appropriate
referrals for treatment. However, inpatient provider performance in recognizing and addressing overweight/obesity or performance variation
among physicians of various training levels and practice settings (hospitalist vs community) has not been reported. Our objective was to describe physician performance in recognizing and managing overweight/
obesity in hospitalized children.
Methods Used: This was a retrospective cross-sectional study of children
ages 2Y18 years hospitalized on the general pediatric service of a tertiary care
children_s hospital between 2009 and 2010. Administrative database height,
weight, and age information were used to calculate BMI percentile using the
CDC BMI calculator. A random sample of patients with BMI at or above
85th% was identified. Physician provider (student, intern, senior, hospitalist,
and community attending) discharge documentation was reviewed to determine the proportion of admitted overweight/obese patients for whom weight
was: 1) recognized as a problem on physical exam 2) included in patient
assessment, and 3) addressed in the care plan.
Summary of Results: Of the 291 patients with BMI percentile at or above
85th percentile whose charts were reviewed, 42% (n=121) were overweight
and 58% (n=170) obese. Patient median age was 6 years (range: 2Y17). BMI
percentile was documented between 0.4% (n=1) and 2.1% (n=5). Overweight
and obesity were recognized only in 1.2% (n=2) and 6.6% (n=16) on physical
exam, 0.4% (n=1) and 4.1% (n=10) on patient assessment, and 0.4% (n=1)
and 2.9% (n=7) in the care plan, respectively.
Conclusions: Overweight/obesity is surprisingly rarely recognized or
addressed during inpatient stays by physician providers regardless of their
training level and work setting. Future studies will explore inpatient provider
perceptions regarding reasons pediatric overweight/obesity is not recognized
or addressed in the pediatric hospital.
Cardiovascular II
Concurrent Session
1:30 PM
Friday, January 28, 2011
221
HEME OXYGENASE-1 IS PROTECTIVE IN AN
EXPERIMENTAL MODEL OF ABDOMINAL AORTIC
ANEURYSM
Azuma J2, Wong RJ1, Maegdefessel L2, Zhao H1, Kalish FS1, Deng A2,
Stevenson DK1, Tsao PS2 1Stanford University School of Medicine,
Stanford, CA and 2Stanford University School of Medicine, Stanford, CA.
150
&
Volume 59, Number 1, January 2011
Purpose of Study: The cytoprotective enzyme, heme oxygenase-1 (HO-1)
is expressed in several vascular cell types, including macrophages. We hypothesized that HO-1 attenuates macrophage activation and inhibits abdominal aortic aneurysm (AAA) formation.
Methods Used: Wildtype (WT, HO-1+/+, n=14) or HO-1 heterozygous
(Het, HO-1+/j, n=13) FVB mice were subjected to aortic elastase perfusion
to induce experimental AAAs. AAA progression was monitored weekly by
ultrasonography. At D7 and 28 post-AAA induction, mice were euthanized
for histological analyses. Expression of inflammatory cytokines and chemokines was assessed by qRT-PCR in thioglycollate-elicited macrophages
from WT and Het mice (n=5 for each group).
Summary of Results: Abdominal aortic diameters were significantly increased in Het compared to WT mice after elastase perfusion. WT mice
exhibited a mean aortic diameter increase of 27.9T2.1% at D7 and 50.5T5.3%
at D28; whereas, Het mice exhibited a mean increase of 45.0T2.4% (pG0.01)
at D7 and 78.7T8.5% (pG0.001) at D28. Het mice also had increased tissue
macrophages at D7 (311T4 vs 180T6 cells per high-power field [hpf],
pG0.001) and at D28 (231T28 vs 43T9 cells/hpf, pG0.001) as compared with
WT mice. Induction of HO activity in Het mice (n=7) by IP injections of
heme (60 Hmol/kg body weight/2 wks) diminished AAA formation to a
level similar to that of WT mice (pG0.001) and resulted in a macrophage
infiltrate reduction of 25% (53T10 cells/hpf, pG0.001) at D28 relative to
untreated Het mice. In addition, thioglycollate-elicited peritoneal macrophages from Het mice exhibited increased expression of the proinflammatory cytokines MCP1, TNF>, IL1A, and IL6 (4.7-, 2.0-, 3.1- and 2.3-fold,
pG0.05, respectively). Meanwhile, mRNA levels of anti-inflammatory
cytokines IL10 and TGFA1 were reduced in Het mice (0.47- and 0.61-fold,
pG0.05, respectively).
Conclusions: In conclusion, induction of HO-1 activity significantly reduces AAA formation in the elastase model of experimental AAA disease. The
underlying mechanism appears in part to be mediated by a reduction of the
local activation of macrophages in the aneurismal site.
222
IDENTIFICATION OF A NOVEL BIOMARKER THAT
DISCRIMINATES HEART FAILURE ETIOLOGY
Maclellan WC, Maclellan R, Horwich T, Fonarow G, LeBlanc J UCLA, Los
Angeles, CA.
Purpose of Study: Despite improvements in the diagnosis and treatment of
CHF, these patients remain at high risk of cardiac mortality and ongoing
morbidity. Presently, the clinician_s ability to diagnose the etiology, determine prognosis or titrate therapy are limited. We sought to identify circulating
biomarkers that might address these limitations.
Methods Used: We analyzed plasma samples collected from 60 CHF
subjects through the Ahmanson-UCLA Cardiomyopathy Center and 44
normal subjects. Non-abundant plasma proteins were enriched by depletion
with an antibody-affinity column. Masses of the eluted plasma proteins were
determined by high-resolution MALDI-TOF mass spectroscopy.
Summary of Results: 794 proteins were detected within the plasma
samples by MALDI TOF-MS. 34 proteins were significantly different between the CHF patients and normal patients, using independent t-tests with a
PG0.05 and adjusted for multiple comparisons using the Bonferoni Correction. Significant proteins were then compared to a panel of clinical variables
(etiology, New York Heart Association (NYHA) classification, left ventricular ejection fraction (LVEF), and outcome to identify potentially important
biomarkers. This analysis demonstrated that the levels of 19 proteins correlated with etiology, 15 with LVEF, 2 with NYHA, and 2 with outcome. Three
of the unknown proteins were tentatively identified based on accurate mass
measurements (less than 0.5 Da error) as isoforms of APO C-III. To further
study APO C-III, we performed ELISA assays on 72 independent subjects.
The study population included 12 aged-matched normal subjects and 60 CHF
subjects, 30 dilated and 30 ischemic, all of them male, to measure circulating
apolipoproteinC-III levels in plasma. ApoC-III levels were elevated in subjects with dilated CHF compared to ischemic etiology and normals
(PG0.00001). ApoC-III levels remained significantly different (PG0.002) after
a multivariate analysis was to account for total cholesterol, total triglycerides
and age.
Conclusions: We have used a novel, high through-put method to identify
potential biomarkers in plasma of CHF patients. Our initial results suggest
that ApoC-III levels correlate with CHF etiology. In the future, we will
confirm these findings in larger study populations.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
223
ZOTAROLIMUS DRUG ELUTING STENTS IN
PERCUTANEOUS REVASCULARIZATION COMPARED
TO 1ST GENERATION DES IN HIGH RISK PATIENTS WITH
LEFT MAIN CORONARY ARTERY DISEASE
Shanmugasudaram M, Ram V, Jayasuria S, Dewar J, Boyella R, Kalra N,
Goldman S, Thai HM University of Arizona/SAVAHCS Medical Center,
Tucson, AZ.
Purpose of Study: Percutaneous coronary intervention (PCI) in patients
with unprotected left main (UPLM) coronary artery disease continues to
evolve following the results of the SYNTAX trial and other studies demonstrating effective and safe outcomes for patients in the PCI cohort. However,
there are limited data on how second generation drug eluting stents (DES)
will affect outcomes in high risk patients undergoing PCI compared to first
generation DES for UPLM.
Methods Used: We retrospectively reviewed 50 patients with UPLM who
underwent PCI with Zotarolimus DES (ZDES) and compared them with
20 patients also with UPLM who underwent PCI with a Sirolimus or
Paclitaxel DES (SPDES) during the same time period and followed them
for 12 months. IVUS guidance was used in all cases. The primary endpoints were: Myocardial infarction (MI), cardiac death and CVA. The
secondary endpoints were: Target vessel revascularization (TVR), ARC
probable stent thrombosis (ST) and contrast use amount as a surrogate for
deliverability.
Summary of Results: Among patients undergoing PCI with ZDES for
UPLM, the major cardiac event (MACE) rate at 12 months was 6 vs 25 % for
SPDES, (p G 0.05). This was accompanied by zero TVR in the ZDES group.
There was a 10% TVR in the SPDES group (pG0.05). Neither stent groups
had any ARC prob ST. The EuroScore was 7.8 + 2.2 among all patients,
there was no difference between ZDES or SPDES groups. In the ZDES
group the contrast used amount was 216.4 + 101 ml vs 413.4 + 160 ml in the
SPDES group, (p G 0.002).
Conclusions: In high risk patients with PCI for UPLM, use of Zotarolimus
DES resulted in a lower 12 month MACE and TVR compared to 1st generation DES. The improved deliverability of the ZDES platform appears to
have a clinical impact, not only in outcomes but also in amount of contrast
use. Our retrospective review suggests that platform deliverability is an independent predictor of clinical outcomes.
224
IMPACT OF RECENT DEVICE PROCEDURES ON THE
INCIDENCE OF CARDIAC DEVICE INFECTION
UNDERGOING EXTRACTION
Moukabary T1, Indik J2, Ott P2, Thal S1 1University of Arizona/SAVAHCS
Medical Center, Tucson, AZ and 2University of Arizona/UMC, Tucson, AZ.
Purpose of Study: The increased demand of device implants and upgrades
based on current guidelines increase the potential for device infections (DI).
Each type of procedure (new implant, device upgrade, device change) may
potentially impact the development of DI. The aim of this study is to identify if having a cardiac device procedure in the previous year increases the
chances of DI compared with patients with no procedures in the previous
year. As a secondary end point we investigated if any type of procedure
carries a higher risk of DI.
Methods Used: Retrospective analysis of the device extraction procedures
(DE) performed at our institutions between January 2000 and December
2009 was conducted. The following data was collected: age of the patient,
left ventricular ejection fraction (LVEF), history of diabetes, COPD or
Creatinine values 9 2 (CR) at the time of DE, type of DI that resulted in DE,
type of procedure, if any, performed in the previous year to the DE. Categorical variables were compared with a Chi square method. Data was
expressed in mean TSD for continues variables and N (%) for categorical
variables.
Summary of Results: 72 patients underwent DE due to DI: Age 69 T12.5
years, males 64 (89%), LVEF 58 T 36 %; diabetes 17 (23.9%); COPD
6 (8.5%); CR 6 (8.5%); 42 patients (58.3%) had solely pocket infection and
30 (41.7%) endocarditis. Of the entire cohort, 43 (59.7%) had a procedure
performed in the previous year compared with 29(40.3%) with no procedure
done (pG0.001). Of the patients who underwent a procedure the most prevalent was device upgrade 21 (29.2%). This showed to be significantly more
common than device change (13 (18.1%); p=0.01) and new implants
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
(9 (12.5%); p=0.03). There was no difference between device change and
new implants (p=NS).
Conclusions: Device procedures within the previous year carries a significant risk of device system infection. Among such procedures, device upgrade
has the highest prevalence, and there is no difference between new implants
and device changes. Prospective data will be necessary to confirm our
findings.
225
CARDIOMYOPATHY FACTORS IN DUCHENNE MUSCULAR
DYSTROPHY
Gray A1,2, Meaney F1, Andrews JG1, Cunniff CM1, Barber BJ1 1University
of Arizona College of Medicine, Tucson, AZ and 2University of Arizona
College of Medicine, Tucson, AZ.
Purpose of Study: Duchenne Muscular Dystrophy (DMD) is characterized by progressive deterioration of both skeletal and cardiac muscle. This
X-linked disorder is caused by mutations in dystrophin, one of the largest
genes identified. Cardiomyopathy (CM ) is one of the leading causes of
death in DMD. CM treatment options are available and therefore early detection is important. Identifying genetic markers associated with early onset
CM could predict which patients require early intervention. Steroids have
become a widely accepted treatment to preserve skeletal muscle strength
for DMD patients, and previous studies have suggested a cardioprotective
role for steroids. The two objectives of this DMD study were to investigate:
1) genetic markers of CM and 2) the effect of steroids on CM onset.
Methods Used: Data were collected through the Muscular Dystrophy
Surveillance, Tracking and Research Network (MD STARnet), a populationbased surveillance program funded by the Centers for Disease Control and
Prevention with five participating sites (Arizona, Colorado, Georgia, Iowa,
and western New York). Echocardiographic (echo), steroid usage, and genetic
data on DMD patients born from 1982-2001 were analyzed. CM was defined
via echo parameters of systolic function: Shortening Fraction G28% and/or
Ejection Fraction G55%.
Summary of Results: The mean age of CM onset was 14.9 years (n=191).
The mean age of steroid initiation was 7.6 years and averaged 61.8 months of
treatment (n=90). The age of CM onset had a significant positive correlation
(r = .45, p G .001) with the number of months treated with steroids. Cases
with a dystrophin point mutation had an onset of CM 22 months earlier
compared to cases with dystrophin deletions or duplications (p = .027).
Conclusions: The significant findings of this MD STARnet study were:
1) dystrophin point mutations are associated with earlier onset of CM and
2) steroid treatment delays the onset of CM in DMD.
226
THE EFFECT OF IVUS GUIDANCE ON CLINICAL
OUTCOMES IN PERCUTANEOUS REVASCULARIZATION
COMPARED TO CORONARY ARTERY BYPASS GRAFTING
AMONG HIGH RISK PATIENTS WITH LEFT MAIN
CORONARY ARTERY DISEASE
Shanmugasudaram M, Ram V, Jayasuria S, Dewar J, Nguyen J, Boyella R,
Goldman S, Thai HM University of Arizona/SAVAHCS Medical Center,
Tucson, AZ.
Purpose of Study: Percutaneous coronary intervention (PCI) in patients
with unprotected left main (UPLM) coronary artery disease continues to
evolve following the results of the SYNTAX trial and other studies demonstrating effective and safe outcomes for intermediate risk patients undergoing
PCI with drug eluting stents (DES). However, there are limited data on how
intravascular ultrasound (IVUS) guidance combined with DES will affect
outcomes in high risk patients undergoing PCI UPLM.
Methods Used: We retrospectively reviewed 119 patients with UPLM who
underwent PCI with DES and mandatory IVUS guidance against 50 patients
with UPLM who underwent CABG surgery during a 3 year period and
followed them for 12 months. The patients were age and gender matched, it
was not possible to perform a propensity risk evaluation, since the CABG
patients were of low to intermediate risks. The primary endpoints were:
(Major adverse cardiac events = Myocardial infarction (MI), cardiac death
and target vessel revascularization (TVR). The secondary endpoints were:
TVR and graft occlusion/or ARC probable stent thrombosis (ST). Logistic
regression analysis was used to model predictors of MACE.
151
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Summary of Results: Among patients undergoing PCI vs CABG for
UPLM, the major cardiac event (MACE) rate at 12 months was (11.8 vs
12.6 %, p = NS). The TVR rate at 12 months was (3.3 vs 4.2%, p = NS) There
was no graft occlusion and/or ST by either groups. There was a significant
difference in EuroScore in the PCI vs CABG cohort (8.4 + 2.2 vs 3.7 + 1.2,
p G 0.05) demonstrating a lower risk group of patients who underwent
CABG revascularization. Presence of EuroScore 9 6 is associated with an
MACE odds ratio (OR) of 7.9 (p G 0.005) in this study population.
Conclusions: Despite having a higher EuroScore, which was significantly
predictive of MACE, patients undergoing PCI with IVUS guidance for
UPLM had a similar clinical event rate compared to low-intermediate risk
patients undergoing CABG at 12 months. The use of DES and IVUS guidance possibly improved the efficacy and safety outcomes. Our retrospective
review lends credence to a future randomized trial incorporating both components in the design.
227
CHEST PAIN AND ST ELEVATION: THE BRUGADA
ALTERNATIVE (CASE REPORT)
Lyons M1,2, Ratliff D1,2 1University of New Mexico, Albuquerque, NM and
2
Albuquerque Veteran’s Hospital, Albuquerque, NM.
Case Report: In the context of ST elevation without evidence of coronary
artery disease but family history of sudden cardiac death, Brugada Syndrome
must be considered.
A 56-year-old male with a past medical history of depression and tobacco
abuse but no cardiac history presented at the Veteran_s Hospital Emergency
room with a chief complaint of chest pain. The patient reported that the
pain was pressure-like in quality and radiated to his left arm. He described
palpitations, shortness of breath and a feeling of Bsomething was wrong.[
The patient was in his yard when the pain began: it lasted 1Y2 minutes during
the first episode and approximately 5 minutes during the second episode.
In the ambulance the patient had ST elevation per monitors. The patient
was given aspirin. The on call team was alerted. In the ED, an ECG was
performed showing ST elevations in leads V1 and V2. Despite his objective
findings, the patient was in no acute distress with stable vital signs. He was
taken directly for a left heart catherization. The patient was found to have
normal coronaries and ejection fraction. The patient was admitted to the
hospital and started on standard medical therapy.
Because of a combination of negative troponins, completely clear coronary arteries and the transient nature of his ST elevations without coronary
artery spasm, the patient was suspected to have Brugada syndrome. He was
discharged and sent to an electrophysiologist where an ICD was placed for
prevention of cardiac death.
Brugada syndrome, first described in 1989, is characterized by particular subsets of ST elevation in the context of a distinct clinical history. The
ST elevation found in Brugada Syndrome is thought to be the result of a
genetic abnormality of the sodium channels of the epicardium. Brugada is
associated with sudden cardiac death, unexplained syncope, sudden ventricular and atrial arrhythmias. The only proven treatment for patients with
symptomatic Brugada syndrome or Brugada pattern with inducible tachycardia is ICD placement. In patients with unexplained arrhythmias and
chest pain, every clinician should add Brugada to the running differential in
his/her head alongside Prinzmetal_s Angina, pericarditis and electrolyte
imbalances.
228
TAKOTSUBO CARDIOMYOPATHY OCCURRING 14 DAYS
AFTER CEREBRAL ANEURYSM RUPTURE
Ashok Kumar M1, Menon N1, Nakaji P3, Sue R2 1St. Joseph’s Hospital,
Phoenix, AZ; 2St. Joseph’s Hospital, Phoenix, AZ and 3St. Joseph’s Hospital,
Phoenix, AZ.
Case Report: Takotsubo cardiomyopathy can present as a life threatening
cardiac event that involves left ventricular apical akinesis, mimicking acute
coronary syndrome (ACS) in patients with an emotional precipitant. It has
also been noted to be associated with subarachnoid hemorrhages and can
present as severe left ventricular dysfunction with cardiogenic shock in this
patient population. The pathogenesis and temporal relationship between these
acute neurological events and Takotsubo cardiomyopathy is not well understood. We present a 62 year old female with no previous history of coronary
152
&
Volume 59, Number 1, January 2011
artery disease or coronary risk factors who developed cardiogenic shock two
weeks after being admitted with a subarachnoid hemorrhage. The echocardiogram showed classic features of Takotsubo cardiomyopathy, including
apical ballooning with basal hyperkinesis. This case is notable for the very
late presentation of this cardiac syndrome after the offending neurological
event. To our knowledge, this is the first reported case of Takotsubo cardiomyopathy presenting so late after a subarachnoid hemorrhage.
229
ASPERGILLUS ENDOCARDITIS OF THE LEFT VENTRICLE
IN A RENAL TRANSPLANT PATIENT: A CASE REPORT
Randhawa T, Taylor R University of New Mexico Medical Center,
Albuquerque, NM, NM.
Case Report: Aspergillus causes an opportunistic infection with a high
rate of dissemination and poor overall outcome especially in immunocompromised patients. In transplant recipients, intense immunosuppressive
therapy to prevent allograft rejection poses a major risk factor for Aspergillosis. In transplant recipients, Aspergillus can affect virtually any organ with
mortality from endocarditis approaching 70%, despite cardiac surgery and
antifungal therapy. We present the case of a 65 year old male who presented
to the hospital after syncope and left sided weakness. Head CT demonstrated
left frontal lobe infarct and echocardiogram showed a left ventricle multilobe mass which was mobile and attached to the posterior basal wall of the
endocardium by a stalk. Histopathology of the resected mass confirmed
Aspergillus endocarditis. Hospital admission was further complicated by
disseminated infection including endophthalmitis, complete heart block,
meningitis and respiratory failure despite resection of the mass and aggressive antifungal medical therapy. Valve replacement or vegetectomy should
be carried out in all patients. The value of combination therapy with antifungal agents is uncertain, however consideration should be given to use a
second agent in addition to liposomal amphotericin.
230
ADULT CONGENITAL PULMONARY STENOSIS. CASE
REPORT AND LITERATURE REVIEW
Alomari IB, Ricciardi MJ UNM, Albuquerque, NM.
Case Report: While pulmonic valve stenosis (PS) is a relatively common
congenital heart condition, it is typically diagnosed and treated in the first
few years of life. We present a relatively rare case of a 21 year old female
with severe PS who underwent successful balloon pulmonic valvuloplasty
(BPV). This 21 y/o Mexican female complained of long-standing shortness of breath and cyanosis. On presentation she was in no distress with BP
142/92, HR 64, and room air O2 Sat 65% (80% on 6LNC). There was
mild cyanosis of lips and digital clubbing. Cardiac auscultation revealed a
4/6 harsh late-peaking systolic murmur at the LUSB, with attenuation of P2.
Lungs sounds were normal. No lower extremity edema. CXR revealed cardiomegaly. TTE revealed severe right ventricular hypertrophy (RVH) with
moderate dilation, moderate pulmonary valve leaflet thickening, and mean
pulmonic valve gradient of 54 mm Hg. A patent foramen ovale (PFO) was
present. She was treated with beta blocker(BB) and referred for cardiac
catheterization and BPV where the hemodynamic findings were confirmed
and shunt study revealed a right to left shunt with Qp 2.7 and Qs 5.5 L/min.
Successful BPV was performed with resultant gradient of 21 mm Hg. Adult
congenital PS is relatively an uncommon disease. Clinical presentation varies
from mild exertional dyspnea to signs and symptoms of right heart failure
depending on the severity of obstruction and the degree of myocardial
compensation. Our patient was unique in that she presented very late despite
severe stenosis and symptoms with right to left shunting through a PFO.
Indication for BPV is similar to surgical valvotomy; namely peak to peak
gradient greater than 50 mmHg. Depending on the degree of RVH, there is
a risk of hyper dynamic contractility and subsequent dynamic RV outflow
obstruction once the obstruction is relieved, a complication called suicide
right ventricle. This may be ameliorated by pre-procedure BB therapy. In
conclusion, severe congenital PS is rarely seen in adults especially in the US
as most cases are corrected early in life. Adult BPV appears to be equally
effective in adults as children and is a viable alternative to surgical valvotomy
in most patients. Reserved for those with gradients greater than 50mmHg,
complications are few. Suicide RV is one such complication that required
special attention.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
231
OVER-THE-COUNTER ENERGY DRINK CONSUMPTION
AND ACUTE MYOCARDIAL INFARCTION: A CASE REPORT
Martinez JD, Sheldon M, Ricciardi MJ University of New Mexico Health
Sciences Center, Albuquerque, NM.
Case Report: Over-the-counter (OTC) energy drinks purportedly enhance
cognitive and physical function in those that use them. Studies have shown
that their consumption results in endothelial dysfunction and platelet aggregation in otherwise healthy subjects. To date, there have been no published reports suggesting adverse clinical events associated with OTC energy
drink consumption. We report a case of a 23 year old healthy male with no
risk factors for premature coronary disease and negative urine toxicology
screening, who presented with an acute inferior myocardial infarction with
heavy intracoronary thrombus burden following recent use of OTC energy
drinks.
Gastroenterology and Hepatology
Concurrent Session
1:30 PM
Friday, January 28, 2011
232
INTERLEUKIN-6 INCREASES INTESTINAL EPITHELIAL
SMALL MOLECULE PERMEABILITY THROUGH
INCREASED CLAUDIN-2 EXPRESSION
Hashimi M, Boivin M, Ye D, Al-Sadi R, Ma T University of New Mexico
Hospital, Albuquerque, NM.
Purpose of Study: Interleukin-6 (IL-6) is a potent pro-inflammatory cytokine that mediates prolonged inflammatory states, such as sepsis, which can
lead to a defective intestinal epithelial barrier. Data regarding the effect of
IL-6 on epithelial permeability remains conflicting. We have previously demonstrated that IL-6 causes a significant increase in epithelial tight junction
permeability as assessed by trans-epithelial resistance (TER) in intestinal epithelial monolayers. We have also demonstrated a role for the MAP kinase
pathway, JNK, in mediating this effect. The major objective of our subsequent
studies is to determine the mechanism through which IL-6 increases intestinal
permeability by focusing on changes in tight junction protein expression.
Methods Used: The effect of IL-6 on intestinal epithelial barrier function
was examined on filter grown Caco-2 cells. Epithelial Monolayer permeability was assessed by apical to basal transit of radio-labeled paracellular
probes of different sizes (urea È2.5 Angstroms, inulin È8 Angstroms). Tight
junction protein expression was analyzed by Western blotting.
Summary of Results: IL-6 increased the permeability of small molecules
such as urea but not the larger probe inulin. IL-6 caused a significant increase
in Claudin-2 expression but did not effect expression of other tight junction
proteins including occludin, ZO-1, MLCK or Claudins 1 and 4. SiRNA to
Claudin-2 blocked the IL-6 induced drop in TER and increase in urea permeability. The JNK inhibitor SP600125 (previously described to prevent the
IL-6 induced decrease in TER) inhibited both the IL-6 induced increase in
urea permeability and the increased expression of Claudin-2.
Conclusions: IL-6 increases the permeability of intestinal epithelial monolayers to small molecules but not large ones. Our studies suggest this is
mediated by a JNK-mediated increase in Claudin-2 expression.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
233
LIPOPOLYSACCHARIDE EFFECT ON INTESTINAL TIGHT
JUNCTION PERMEABILITY: A KEY ROLE FOR MYOSIN
LIGHT CHAIN KINASE
Alhmoud TZ1, Al-Sadi R1, Ma T1,2 1University of New Mexico, Albuquerque,
NM and 2Albuquerque Veterans Affairs Medical Center., Albuquerque, NM.
Purpose of Study: Lipopolysaccharide (LPS) is a major part of gram
negative bacteria outer membrane. It can illicit strong immune responses
in-vivo and in-vitro. Intestinal epithelial tight junctions (TJs) are the apical
most junctional complexes that act as functional and structural barrier against
permeation of harmful luminal antigens. Defective TJ barrier is a key factor in
inflammatory conditions of the gut, leading to increased permeation of noxious substances which promote intestinal inflammation. It has been shown
previously that myosin light chain kinase (MLCK) plays an important role in
regulation of intestinal epithelial TJ barrier function through phosphorylation of myosin light chain (MLC) leading to contraction of perijunctional
acto-myosin filaments, opening of the TJ barrier and increase permeability.
However, the effect of LPS on intestinal epithelial TJ permeability at
physiologically relevant concentrations and its mechanisms of action remain unknown.Our aims in this study were to determine the effect of LPS
on intestinal epithelial TJ permeability and to delineate the intracellular
mechanisms involved.
Methods Used: Filter-grown Caco-2 intestinal epithelial monolayers were
used as an in-vitro intestinal epithelial model system and following assays
were used:
1) Trans-epithelial resistance (TER);
2) Western blot analysis;
3) Intestinal permeability.
Summary of Results: 1) LPS treatment at physiologically relevant concentrations (0.1Y1 ng/ml) caused a time and dose-dependent decrease in
trans-epithelial resistance (TER) in Caco-2 monolayers. 2) Conversely, LPS
treatment resulted in a time-dependent increase in permeability to paracellular
marker inulin. 3) The LPS-induced increase in Caco-2 TJ permeability correlated with a time-dependent increase in myosin light chain kinase (MLCK)
protein expression. 4) SiRNA-induced knock-down of MLCK resulted in a
near complete depletion of MLCK protein expression. 5) Silencing MLCK
expression by siRNA transfection prevented the LPS-induced increase in
Caco-2 intestinal TJ permeability.
Conclusions: In conclusion, our data demonstrated that LPS caused an
increase in intestinal TJ permeability and that was mediated by an increase in
MLCK protein expression.
234
ROLE OF P38 MITOGEN ACTIVATED PROTIEN KINASE ON
THE IL-1B-INDUCED INCREASE IN INTESTINAL TIGHT
JUNCTION PERMEABILITY
Alhmoud TZ1, Al-Sadi R 1, Ma T1,2 1University of New Mexico,
Albuquerque, NM and 2Albuquerque Veterans Affairs Medical Center,
Albuquerque, NM.
Purpose of Study: Interleukin-1 Beta (IL-1B) is a prototypical proinflammatory cytokine that plays an important role in the inflammatory conditions of the gut. It has been shown that IL-1B caused a marked increase in
intestinal epithelial tight junction permeability.
The IL-1B induced increase in epithelial TJ permeability has been
postulated to be an important pathogenic factor contributing to intestinal
inflammation.
However; the signaling pathways involved in the IL-1B-induced increase
in TJ permeability remain unclear. Our aim in this study was to delineate the
role of p38 mitogen-activated protein kinase (MAPK) in the IL-1B-induced
increase in TJ permeability.
Methods Used: Caco-2 intestinal epithelial monolayes were used as an
in-vitro intestinal epithelial model system,and following assays were used:
1) Trans-epithelial resistance (TER);
2) Western blot analysis.
Summary of Results: Results: 1) IL-1B treatment (10ng/ml) caused a time
and dose-dependent decrease in trans-epithelial resistance (TER) in Caco-2
monolayers. 2) The IL-1B induced decrease in Caco-2 TJ resistance was
preceded by activation of p38 kinase. 3) Inhibition of p38 by SB-203580
(10 mM) prevented the IL-1B-induced drop in Caco-2 TER. 4) To further
validate these results, p38 kinase was selectively knocked-down by siRNA
153
Western Regional Meeting Abstracts
Journal of Investigative Medicine
p38 (small interfering RNA which is a small double-stranded RNA molecule used to silence gene expression of a specific protein.); siRNA-induced
depletion of p38 kinase prevented the IL-1B-induced drop in Caco-2 TER. 5)
Activation of p38 pathway caused a down-stream activation of nuclear transcription factor (ATF-2). Inhibiting ATF-2 by siRNA-induced knock down
prevented the IL1B-induced drop in Caco-2 TER.
Conclusions: In conclusion, this study demonstrated that IL-1B-induced
increase in intestinal TJ permeability is mediated by activation of p38 signaling pathway leading to the activation of ATF2 nuclear transcription factor.
235
WILSON DISEASE: ABNORMAL METHIONINE
METABOLISM IN PATHOGENESIS OF LIVER DAMAGE
Dolatshahi M, Jose S, Halsted C, Medici V University of California, Davis,
Davis, CA.
Purpose of Study: Wilson disease (WD) is a recessive disorder of copper
accumulation with established genetic background but unclear pathogenesis.
Aberrant hepatic methionine metabolism plays a role in the pathogenesis of
liver injury by regulating homocysteine (HCY) and S-adenosylhomocysteine
(SAH), which induce endoplasmic reticulum (ER) stress and consequent
induction of lipogenic pathways. We hypothesized that abnormal methionine
metabolism is involved in the pathogenesis of hepatic steatosis in WD.
Methods Used: Livers were removed from Jackson toxic milk mice (tx-j), a
model of WD, and C3H control mice fed the same diet at 16, 20, and 24 weeks
of age. Profiles of selected hepatic methionine metabolites in the tx-j mice were
compared to those of the wild type mice at the three ages and were correlated
with histopathology, triglyceride accumulation, and activation of ER stress,
apoptosis, and lipogenesis markers.
Summary of Results: At 20 and 24 weeks, the tx-j mice showed greater
hepatic triglyceride levels and higher cytoplasmic lipid droplets in hepatocytes compared to the controls. SAH hydrolase (SAHH) mRNA expression,
protein levels, and activity were significantly lower in the tx-j mice at all ages
and correlated negatively with hepatic copper levels, which were significantly
higher in all tx-j mouse age groups. The transcript and protein levels of the ER
stress marker glucose related protein 78 (GRP78), the apoptosis marker growth
arrest DNA damage inducible gene 153 (GADD 153), and the lipogenesis
transcription factor sterol regulatory element binding protein 1c (SREBP-1c)
were increased at all age points in the tx-j mice. Plasma HCY levels were elevated at 20 weeks and correlated with hepatic triglyceride levels.
Conclusions: Reduced hepatic SAHH expression was associated with increased SAH and subsequently with increased plasma HCY as well as the
activation of ER stress lipogenic pathway gene expression, which were paralleled by the progression of hepatic damage. These findings support the
hypothesis that aberrant methionine metabolism plays an important role in
liver damage pathogenesis in WD.
236
PREVALENCE OF PEDIATRIC INFLAMMATORY BOWEL
DISEASE AMONG HISPANICS IN AN URBAN ACADEMIC
CENTER IN THE SOUTHWEST UNITED STATES
Adair JG, Gremse DA, Scherr RL University of Nevada School of Medicine,
Las Vegas, NV.
Purpose of Study: Inflammatory bowel diseases (IBD), ulcerative colitis
(UC) and Crohn disease (CD), affect approximately 2 million Americans.
While it has been historically thought to be a disease affecting Caucasians
it is now apparent that other ethnic groups are also affected. The Hispanic
population is increasing in the United States; therefore, this study was done
to determine the prevalence of IBD among ethnicities in an urban pediatric
gastroenterology clinic in the southwestern United States.
Methods Used: A retrospective medical record review was performed on
1424 pediatric patients between the ages of 1 and 18 years of age who were
seen at an urban academic pediatric gastroenterology center in Las Vegas
during the years 2004Y2010. The race and ethnicity of all inpatient and
outpatient cases were identified. Groups included African American non
Hispanic (AA), Caucasian non Hispanic (C), Hispanic (H), Asian/other (O)
or unknown. In addition, any patient with the diagnosis of CD, UC, proctitis,
nonspecific colitis, IBD, or colitis were evaluated for radiographic, endoscopic, histologic and clinical course consistent with the diagnosis of IBD.
The prevalence of IBD among race/ethnic groups was compared using a
154
&
Volume 59, Number 1, January 2011
two sample t- test. A p value of G 0.05 was defined as being statistically
significant.
Summary of Results: The patient race/ethnicity distribution of all records reviewed was as follows: 11.6% AA, 42.8% C, 39.2% H, 3.2% O, 3.2%
unknown. In comparison, the Clark County School District has a distribution of 41% H, 12% AA, 42% C, and 5% O. Forty-three of the 1424 patients
were diagnosed with IBD, 29 CD and 14 UC. The prevalence of IBD by race
was 8/558 H (1.4%), 8/165 AA (4.8%), and 27/610 C (4.4%). The prevalence
of IBD between C (4.4%) and AA (4.8%) was similar (p=0.82). However,
the prevalence of IBD was significantly higher in each of these races compared to Hispanics (C vs. H, p=0.0027, AA vs. H, p=0.0088).
Conclusions: In this pediatric patient population, Hispanics have decreased prevalence of IBD compared to Caucasians and African Americans.
Although genetics contribute to the development of IBD, the role of environmental and dietary factors cannot be overlooked.
237
TREATMENT OF CHRONIC HEPATITIS B PATIENTS
ACCORDING TO CURRENT GUIDELINES: A
COMMUNITY-BASED STUDY
Zhang S1,2, Ristau J2, Trinh HN3,2, Garcia RT3,2, Nguyen H3, Levitt BS3,
Nguyen MH4,2 1UCLA School of Medicine, Los Angeles, CA; 2Pacific Health
Foundation, San Jose, CA; 3SJGI, San Jose, CA and 4Stanford University
Medical Center, Palo Alto, CA.
Purpose of Study: Prior studies have found that a significant proportion of
chronic hepatitis B (CHB) patients do not receive antiviral therapy (AVT).
Our goals were to characterize CHB patients by treatment eligibility and
actual treatment status by current published guidelines and to examine reasons for the lack of AVT in patients who are treatment eligible.
Methods Used: We conducted a retrospective study of patients who were
first evaluated for CHB at one of two community gastroenterology clinics
between April 2007 and February 2009. Using criteria published by the
American Association for the Study of Liver Diseases (AASLD) in 2009
and a panel of U.S. hepatologists (US Panel) in 2008, treatment eligibility
was determined for patients using clinical and laboratory data from the
first six months of presentation. Treatment status was followed in the patients
who were found to be treatment eligible and reasons for non-treatment were
identified.
Summary of Results: A total of 612 consecutive CHB patients were included in the analysis. Mean age was 44T13 years, 54% were male, and
almost all were Asian (99%). About half of the patients (51%) were treatment eligible by US Panel guidelines. Of these patients, 47% additionally
met AASLD guidelines. The latter were more likely to be younger, hepatitis B e antigen positive, and had higher median alanine aminotransferase level (ALT) and HBV DNA levels. Overall, treatment was initiated in
50% of eligible patients: 29% of US Panel-eligible-only patients and 72% of
AASLD-eligible-patients. On multivariate analysis, independent predictors
for actual treatment initiation were higher ALT in AASLD-eligible-patients
and higher ALT and older age in US Panel-eligible-only patients. The leading reason for non-treatment was further observation as desired by the
physicians.
Conclusions: Approximately half of treatment eligible patients did not
receive antiviral therapy for CHB as recommended by either AASLD or
US Panel guidelines. Substantial undertreatment of CHB is observed especially in patients eligible only by US Panel guideline and should be further
examined.
238
ROLE OF ANTIMICROBIAL PEPTIDES IN ESOPHAGITIS
Robinson ZD1, Masterson J1,2,3, Hosford L1, Fillon S1,2,3, Furuta GT1,2,3
1
University of Colorado, Denver, CO; 2The Children’s Hospital, Denver, CO
and 3Digestive Health Institute, Denver, CO.
Purpose of Study: Esophagitis is caused by exposure of the epithelia to
allergens (eosinophilic esophagitis-EoE) or acid (gastroesophageal reflux
disease-GERD) but the exact pathogenetic mechanisms of these diseases are
unknown. Recent data supports a potential role for microbial flora in the
initiation or perpetuation of esophagitis. We hypothesize that dysregulated
innate defense molecules (defensins) of esophageal epithelia contribute to
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
esophagitis. This study aimed to measure defensin expression in patients with
esophagitis and in esophageal epithelial cells of an in vitro model of eosinophilic inflammation.
Methods Used: Defensin (hAD1 and 3) expression from esophageal biopsies
(EoE, GERD and normal) was measured by qRT-PCR. Epithelia hAD1 and 3
expression of an in vitro model of esophagitis [esophageal epithelia-HET-1A
cell line and human peripheral eosinophils-IL-5 (100pg/ml) and GMCSF
(100pg/ml)] were measured via qRT-PCR.
Summary of Results: Esophageal biopsies from EoE patients have decreased expression of hAD1 compared to normal (0.19T0.22 vs 1.0T0.61,
respectively; p=0.019). EoE patients treated with topical steroids have similar
hAD1 expression levels compared to normal (1.0T0.80 vs 1.0T0.61 respectively; p=0.56). HAD3 expression in biopsies from EoE patients is significantly decreased compared to normal (0.15T0.21 vs 1.0T0.65, respectively;
p=0.03). HAD3 expression in EoE patients treated with topical steroids trends
but is not significantly reduced compared to normal (0.21T0.14 vs 1.0T0.65
respectively; p=0.06). HAD3 expression in biopsies from GERD patients and
GERD patients treated with proton pump inhibitors is significantly decreased
compared to normal (0.10T0.07 vs 1.0T0.65 and 0.17T0.22 vs 1.0T0.65;
p=0.03 and p=0.03, respectively). HET cell hAD1 and 3 expression are
significantly reduced by IL-5 and GMCSF compared to HET cells cultured in
media alone (0.71T0.12 vs 1.05T0.11 and 0.39T0.14 vs 0.95T0.24; p=0.03
and p=0.01, respectively). HET cells cultured with IL-5, GMCSF, and
eosinophils have a trend towards decreased hAD1 expression compared to
controls without eosinophils (0.77T0.08 vs 1.24T0.22; p=0.07, respectively).
Conclusions: Defensin expression is decreased in esophagitis. We speculate
that dysregulated defensin expression contributes to esophagitis.
Western Regional Meeting Abstracts
Purpose of Study: To delineate the intra cellular signaling pathway that
mediates endotoxin induced increase in intestinal permeability using filter
grown caco-2 monolayer as an in vitro model system.
Methods Used:
1. Filter grown Caco-2 intestinal epithelial monolayer were used as an in vitro
intestinal epithelial model system
2. Intestinal permeability was assessed by measuring Trans epithelial resistance (TER).
3. Western blot analysis.
4. ELISA induced assay was assessed to determine the activation of ELK1(transcription factor).
Summary of Results:
1. LPS at physiologically relevant concentrations of 0.3 ng/ml caused a time
dependent drop in TER.
2. LPS at physiological concentrations of 0.3 ng/ml caused a time dependent increase in P38 kinase activation as determined by P38 kinase
phosphorylation.
3. Inhibition of P38 kinase using an inhibitor prevented the LPS induced drop in TER.
4. LPS treatment resulted in time dependent activation of P38 kinase ELK-1.
5. Inhibiting ELK-1 expression by SiRNA prevented the LPS induced drop in
Caco-2 Trans epithelial resistance.
Conclusions: LPS at physiological concentration caused an increase in
Caco-2 Tight Junction permeability
LPS induced increase in intestinal permeability was mediated by P38
kinase activation of ELK-1 transcription factor.
240
EPIDEMIOLOGY OF BILIARY ATRESIA IN NEW MEXICO
239
P38 KINASE PATHWAY MEDIATES ENDOTOXIN INDUCED
INCREASE IN INTESTINAL EPITHELIAL TIGHT JUNCTION
PERMEABILITY
Naqvi MR1,2, Sadi RA2, Ma T2,1 1University of New Mexico, Albuquerque,
NM and 2UNM, Albuquerque, NM.
Rose R, McLoughlin L University of New Mexico, Albuquerque, NM.
Purpose of Study: Biliary Atresia is a disease that is defined as a complete
obstruction of the lumen of the extrahepatic biliary tree and it is the most
common cause of death from liver disease in early childhood. The purpose of
our study was to investigate the possible existence of seasonal, temporal,
community size or regional influences in the occurrence of Biliary Atresia
in New Mexico.
Methods Used: All patients with the diagnosis of Biliary Atresia who were
born in New Mexico in the years 2003Y2007 were included in this study. The
data was collected from the University of New Mexico as well as Presbyterian
Hospital, which are the only facilities in New Mexico that can make the
diagnosis. Each patient_s season and year of birth, regional location, and
community (urban vs. rural) were evaluated in this analysis. Statistical
analysis was done utilizing the Chi Square Goodness of Fit model.
Summary of Results: We identified 14 patients who met the inclusion criteria. Our data did not show any significance in seasonality, community or
regional location of birth. While the years as a whole did not show any statistical
significance, the year 2004 did show a significant increase with a p-value G.05.
Conclusions: We conclude that there are not statistically significant seasonal, community or regional based variations in the incidence of biliary
atresia in New Mexico. Also, while there is no significance over the time
period as a whole, 2004 was shown to have a significant increase in the cases
of Biliary Atresia. Our data can be used to further elucidate the diagnosis of
Biliary Atresia and its possible contributing factors.
241
SPONTANEOUS RUPTURE OF HEPATIC HEMANGIOMA
IN A NEONATE - SURVIVAL WITH NON-OPERATIVE
MANAGEMENT
Drop in RR after Rx with LPS
phosporylation of P38 kinase
* 2011 The American Federation for Medical Research
Ghuman A1, Schreiber R2,1, Butterworth S1,3 1University of British
Columbia, Vancouver, BC, Canada; 2BC Children’s Hospital, Vancouver,
BC, Canada and 3BC Children’s Hospital, Vancouver, BC, Canada.
Background: Hepatic hemangioma is the most common benign pediatric
tumor of the liver. Spontaneous rupture of these tumors is extremely rare and
carries a very high mortality rate.
Case Report: A 7 day old term male infant presented to the Emergency
Room with jaundice, lethargy, increased work of breathing and a tense abdomen in severe hypovolemic shock. He was intubated, fluid resuscitated and
received a blood transfusion. On admission his hemoglobin was 27. An
abdominal ultrasound demonstrated a large right hepatic subcapsular hematoma, massive hemoperitoneum and numerous additional focal echogenic
155
Western Regional Meeting Abstracts
Journal of Investigative Medicine
lesions in his liver. An abdominal CT scan confirmed these findings and
further characterized the lesions to be most consistent with hepatic hemangioma. The patient was placed on high dose IV steroids and repeat ultrasound
performed 7 days later found the hemoperitoneum had resolved and the
subcapsular lesion persisted. Given that life-threatening hemorrhage had already occurred, it was felt the risk for recurrent bleeding was substantial and
he underwent selective right hepatic arterial embolization which supplied
the ruptured lesion 17 days after admission. He was discharged home on
3 mg twice daily of prednisolone which was tapered to off over 3 months.
At 19 months he is healthy and developing normally. The last ultrasound
demonstrated multiple lesions still present, but decreased in conspicuity.
Conclusion: We present a rare case in which a patient with spontaneous
rupture of a hepatic hemangioma survived and is thriving today with
no immediate surgical intervention. To our knowledge this is the first reported
neonate to have a ruptured hepatic hemangioma and survive by managing
with resuscitation, steroids and delayed embolization.
Health Care Research II
Concurrent Session
1:30 PM
Friday, January 28, 2011
242
EVALUATING DESIGN ELEMENTS IN COMMONLY VISITED
HYPERTENSION PATIENT EDUCATION WEBSITES
Stratton SR, Dotson A, Kim S University of Washington, Seattle, WA.
Purpose of Study: The quality of the health education sites returned by
hypertension-related Internet search queries is of increasing importance.
Specific web design and user interactivity features have been shown to support learning and retention. This study reviewed systematically the content
presentation and web design features of sites commonly visited by patients
searching for information about hypertension.
Methods Used: Using Google AdWord_s Keyword Tool, we identified 60
commonly used search phrases for hypertension and their monthly search
volume. Based on projections of click-through traffic, we generated a list of
39 unique hypertension patient health education sites with at least 150,000
estimated visits per month. Using the results of previous studies examining
website design features, we developed a site design quality score (SDQS) to
grade sites based on the presence or absence of 12 desirable design elements,
11 interactive features, and 13 content transparency and integrity characteristics. The potential score was 0Y36.
Summary of Results: Monthly search volume for selected phrases ranged
between 673,000 and 2,240,000. Websites had a median design element score
of 4/12, median interactivity score of 3/11, and median transparency and
integrity score of 5/13. The overall median SDQS was 14, range 4Y26. The
median Flesch-Kincaid reading grade level was 8.6, range 3.4Y14. Healthcentered non-profits, governmental, and educational sites (n=9) had a median
SDQS of 17 while commercial sites (n=22) had a median SDQS of 11.5.
There was a poor association between SDQS and estimated site traffic from
Google (r=.197).
Conclusions: Search volume for hypertension is high, and patients use a large
variety of search terms. There is great variation in the design quality of patient
health education websites for hypertension. All websites examined have significant room for improvement in design, interactivity, readability, and/or
transparency as well as in the use of multimedia and user self-assessments. Nonprofit, educational, and governmental sites had a higher median SDQS score
than commercial sites. Google_s search algorithm does not strongly select for
hypertension patient education sites that have higher SDQS scores.
243
ONSITE CLINICS IN SIX WORKPLACES: MEASURES
OF SUCCESS
Raymond LW1,2, Pankowski J2, Burns D2, Cole B2, Lovings T2, Shiflett H2,
Yanni A2, Bowers L2 1Univ of North Carolina, Chapel Hill, Charlotte, NC
and 2Carolinas HealthCare System, Charlotte, NC.
Purpose of Study: Workplace clinics (WPCs) provide onsite care for minor
illnesses, can assist primary clinicians (PCs) in disease management, and can
156
&
Volume 59, Number 1, January 2011
deliver occupational health services. There are few reports of utilization
patterns and financial benefits of WPCs, however. We measured near-term
WPC outcomes at a pipe manufacturer (PM), town hall (TH), lumberyard (LY),
canning facility (CF), metal fabricator (MF), and electric insulator manufacturer (EIM). Co-pays were $3 per visit at TH and zero at other WPCs.
Methods Used: We placed a part-time mid-level clinician (MLC) at each
WPC and recorded visit numbers, diagnoses and referrals in each, and estimated cost savings to employers. We used visit complexity as a surrogate for
the cost which would have been incurred, had the service been rendered
offsite in Charlotte, NC.
Summary of Results: Utilization was high (Table). Leading reasons for
visits included: hypertension and dyslipidemia (each 12%), respiratory and/
or allergy symptoms (11%), and obesity (9%). Hence, most visits were for
conditions handled by the MLC. Referrals were made in 24% of visits, nearly
half (11%) to the worker_s PC. Over-the-counter medicines were advised in
60% of visits, and agents with the lowest co-pay were prescribed in another
30% of visits. Estimated cost savings from the WPC ranged from $249 to
$874 per worker per year.
Conclusions: Employers realized substantial savings from WPCs, ranging
from $103 to $320 for each hour of MLC service. However, our analysis does
not identify the costs billed by PCs, and hence could overstate the actual
savings to an unknown extent.
Demographic and Savings of Onsite Clinics at 6 Worksites
244
BUILDING CAPACITY TO REDUCE DIABETES
DISPARITIES:TRAINING COMMUNITY HEALTH WORKERS
Bouchonville M1, Colleran K1, Kipp B2 1University of New Mexico HSC,
Albuquerque, NM and 2University of New Mexico HSC, Albuquerque, NM.
Purpose of Study: Community health workers (CHWs) participate in many
aspects of heath, through committment to their communities by overcoming
barriers to care:language, resources, culture, and trust. CHWs are ideally situated to help tackle the diabetes epidemic. Unfortunately, CHWs face barriers
to obtaining training including: time, cost, and travel. Using the ECHO model,
we developed a distance training program utilizing case based learning, didactics and resource sharing. We hypothesize that participation in the program
will increase CHW attitudes toward treatment, knowledge, and confidence in
working with diabetics.
Methods Used: Twenty-three subjects were enrolled in the first training.
Subjects completed baseline attitude, knowledge (DKT), and confidence
surveys. They attended a three day skills training session, followed by weekly
participation in tele/video conferences for 6-months. Each session consisted
of a didactic, case presentations by participants which were discussed by the
UNM multidisciplinary diabetes team and participants, Q&A and resource
sharing. Participants completed surveys at the end of the training. Wilcoxon
Signed Rank statistics were used to compare pre and post test results. Regression analyses were also done.
Summary of Results: Twenty-one subjects completed the program. There
were significant improvements in the DKT, 57% correct at baseline versus
71% correct at the end of the study p=0.0002. Confidence in clinical skills
increased significantly from 3.3 at baseline to 4.4 (on a scale of 1Y5)
pG0.0001. Confidence in non clinical skills including education and advocacy increase from 3.62 to 4.29, p=0.0002. Participants demonstrated improved attitudes on the seriousness of diabetes (4.1Y4.39, p=0.04). Regression
analysis demonstrated significant correlation between attitudes of the seriousness of diabetes and the psychological impact of diabetes, the role of
patient autonomy, and participant confidence in clinical skills, r= 0.55,
p=0.01, r=0.57, p=0.007, r=0.46, p=0.36, respectively.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Conclusions: CHW participation in the ECHO distance training program
resulted in significant increases in knowledge, confidence, and attitudes in
diabetes. Future studies are ongoing to determine if the training has a impact
on patient outcomes.
245
AVOIDING THE IMPRESSION OF IMPROPRIETY:
UNDERSTANDING COMMON ERROR TYPES IN
PUBLICATIONS CITED BY RESIDENCY APPLICANTS
Simmons HR1, Kim S2, Zins AM2, Amies-Oelschlager AE2 1University of
Washington, Seattle, WA and 2University of Washington School of Medicine,
Seattle, WA.
Purpose of Study: Multiple studies have demonstrated that many publications reported by applicants to residency are unverifiable when searched.
We sought to investigate what type of errors medical students may make in
reporting a publication that may preclude the ability for a listed publication to
be verified.
Methods Used: We performed a retrospective review of the ERAS applications submitted to the University of Washington OB/GYN residency for the
2008 and 2009 matches. We searched reported peer reviewed journal articles/
abstracts, published and unpublished (submitted/accepted) submitted in
PubMed and exhaustive list of online resources. If still unfound, journal
editors were contacted to inquire about the article in question. Errors were
categorized as: 1) deliberate (insertion of name or article never submitted), 2)
overstatement (incorrectly classifying a publication as peer reviewed, listing
articles where they were not an author, changing the ranked author list, or
citing publications without authors, 3) minor (deleting all other authors, title
change, wrong journal publication name, date, volume, wrong Pub Med ID
number, or failure to list the journal name), and 4) unverifiable (publication
unable to be found despite exhaustive search).
Summary of Results: Of the 937 applicants that 546 applicants (58.2%)
listed a total of 2251 publication entries (mean = 4.12). Of these, 354
applicants listed 999 peer reviewed journal articles/abstracts, of which 751
were reported published and 248 reported unpublished. 150 (42.6%) of
applicants who reported a peer reviewed publications had at least one error
identified or a publication which could not be found. In the 999 publications,
there were 10 deliberate errors (1.0%), 161 overstatement errors (16.1%), and
54 minor errors (5.4%). 157 (15.7%) publications or journals were unverifiable. All publications with a Pub Med ID number listed were able to be
verified.
Conclusions: Over one third of peer reviewed publications listed by medical
students applying in OB-GYN are erroneous in some manner. Students
should be aware of these errors in order to avoid the appearance of impropriety, real or perceived.
246
HEALTH NEEDS AND HEALTHCARE SYSTEM RESPONSES
AMONG VICTIMS OF GENDER-BASED VIOLENCE
IN AFGHANISTAN
Stokes S, Miller E UC Davis School of Medicine, Sacramento, CA.
Purpose of Study: Gender-based violence remains a challenge to promoting women_s health in Afghanistan. This study aims to describe the range
of health needs and heterogeneity of healthcare experiences among Afghan
women exposed to gender-based violence.
Methods Used: In-depth, anonymous face-to-face interviews were conducted with 22 Afghan women ages 18 and older currently receiving services
through a Kabul-based non-governmental organization providing shelter to
victims of human rights abuses. When permitted, the interviews were audiotaped and transcribed. Otherwise, detailed notes were taken during the
interviews. All interviews were coded for key themes related to health and
abuse utilizing a content analysis approach.
Summary of Results: The median age of respondents was 19 years (range
18Y26 years). Over half of women reported histories of physical abuse by
their husband (60%, n=13), 46% by their husband_s family (n=10) and 41%
by members of their own family (n=9). 82% also reported receiving threats of
honor killings (n=19). Almost all women endorsed current symptoms of depression (91%, n=20), and eleven reported attempting suicide in response to
abuse experienced. Nearly all women had received medical care for symptoms
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
associated with their abuse (n=19). Nine women were treated for severe injuries
sustained from physical abuse, nine were hospitalized for attempting suicide
and two were required to undergo virginity examinations by a physician. Only
one woman reported that her healthcare provider attempted to report her abuse
to authorities. All women receiving medical care for symptoms associated with
their abuse denied being offered any mental health counseling (n=19), and none
were informed by their healthcare providers about supportive services for
victims of gender-based violence.
Conclusions: The interviews suggest that Afghan women receiving medical care for health complications associated with gender-based violence
may not be properly identified by healthcare providers and may not be receiving counseling or supportive services through healthcare providers.
Further studies on healthcare responses to victims of gender-based violence
are needed. Training in assessment for abuse during medical evaluations
may assist healthcare providers in Afghanistan to better recognize victims
of gender-based violence and to connect women to support services.
247
SMOKING CESSATION DURING PREGNANCY INCREASES
MOTIVATION TO BREASTFEED
Merritt TA, Amstrong S Loma Linda University, Loma Linda, CA.
Purpose of Study: In the second and third trimesters we showed the video
‘‘When You Smoke Your Baby Smokes’’ to 96 high risk women which
highlighted the adverse effects of tobacco smoke on their fetus and newborn.
Methods Used: The videos and follow up questionnaires were followed by
motivational interviews to the gravida or recently delivered mothers.
Summary of Results: Of 96 pregnant mothers, 92% of 25 self identified
as currently smoking (or smoking within the last year) were enrolled in a
tobacco treatment program which provides an incentive-based, 8 week program
with monitoring using salivary cotinine. Fifteen (60%) remained tobacco free
for 8 weeks and at the time of delivery. Among the 40% who failed to remain
tobacco free, viewing the video prompted mothers to quit for at least one week.
21 mothers (83%) chose to breastfeed during the postpartum period.
Conclusions: After delivery (mean Gestational Age mean 35.2+/j3.1 weeks)
the impact of the program prompted 25% of mothers who smoked early in
pregnancy to change from a state of contemplation to preparation and 65%
to change from contemplation to action. Mothers also disclosed that viewing
the video increased their desire and willingness to breastfeed their infant.
Maternal-fetal attachement and the desire to breastfeed is enhanced among
mother who receive video information regarding the hazards of tobacco
smoking during pregnacy and offered enrollment in an incentive-based
program designed to assist mother to quit smoking. We conclude that video
education regarding the hazards of tobacco smoking is an effective tool to
prompt enrollment in smoking cessation programs and to accelerate stages
of change to quit smoking.
248
PATIENT COMPLIANCE IN A CLINICAL TRIAL OF AN
HERBAL THERAPY FOR TYPE-2 DIABETES
Brueckner E, Martin JT, Nguyen DH, Mendoza N, Nguyen B, Espino R,
Hoettedumlao J, Semenova Y Western University of Health Sciences,
Pomona, CA.
Purpose of Study: Patient compliance is the most critical part of the treatment of chronic diseases. Unfortunately, with diseases like Type-2 Diabetes,
where disease onset is insidious, patients demonstrate poor compliance until it
is too late. Compliance in diabetics taking mainstream glycemic-control drugs
is well studied; however, compliance in diabetics taking herbal therapy has not
been investigated. Our study examines compliance among treatment groups
over the duration of our clinical trial.
Methods Used: 107 Type-2 Diabetic subjects are enrolled in a 6 month,
double-blind, placebo controlled trial. Criteria for participation include
having a HA1c value between 7.0 and 9.0, being age 20Y75, having normal
renal and hepatic function, and having undergone no insulin therapy or
diabetic-regimen drug or dosage changes for at least 3 months before and
throughout the trial. Subjects take 1 capsule of the drug or the placebo by
mouth, once daily for 6 months. On a monthly basis, participants meet with
study coordinators to retrieve more capsules, return remaining capsules and
157
Western Regional Meeting Abstracts
Journal of Investigative Medicine
their daily glucose log. The capsules returned are counted, and these values
are used to measure compliance.
Summary of Results: The fraction of subjects that returned more capsules
than expected (indicating that they missed doses) significantly increased over
the six month trial period. The average number of capsules returned by the
experimental and control groups did not differ significantly (T=1.39, DF=43,
P=.17). However, the variance in number of capsules returned was significantly lower in the experimental group (F=1.94, P=.049).
Conclusions: An explanation for the difference in variance between the
control and experimental group may be related to the daily monitoring of the
glucose levels that the subjects performed. If our herbal therapy has a blood
glucose lowering effect, the subjects will notice the changes in their daily
glucose readings. This awareness among the experimental group provides an
incentive to maintain their initial level of compliance, and explains the relatively small variance. The control group lacks this hypothesized incentive,
and therefore is less likely to maintain a steady compliance levels throughout
the study, leading to relatively large variance.
249
BIOMECHANICAL CHARACTERIZATION OF THE
CARTILAGINOUS ENDPLATE IN THE INFANT SPINE
Farmer RP, Paietta R, Ferguson V, Burger E University of Colorado, School
of Medicine, Aurora, CO.
Purpose of Study: The purpose of this study is to improve our understanding of the interface between mineralized and non-mineralized biological
tissues. This is of great importance to ensure successful prosthetic implant
fixation and to improve our understanding of the etiology of spinal disc
degeneration and developmental disorders. In the spine, bone and cartilage
meet at the osteochondral interface - a region that experiences high shear
forces in normal use. We seek to characterize the gradient of properties from
bone to cartilage within this interface using scanning electron microscopy,
in both backscattered (BSE) and secondary electron (SEM) modes, to visualize the 2-D and 3-D structures of the cartilaginous endplate region in the
human infant spine.
Methods Used: Within this interface, anchoring of the intervertebral disc
(IVD) to the adjacent vertebral bodies by collagen fibrils will be assessed via
immunohistochemistry staining for collagen types I, II, IV, and X. BSE imaging
methods were developed using mature, ovine lumbar spine samples that were
histologically embedded in poly(methylmethacrylate) (PMMA) prior to sectioning and imaging. Additional BSE imaging has also been performed on a
human fetal (37 weeks gestational age) lumbar spine L3-L5. A second ovine
lumbar vertebra is currently being in order to image the mineralized surface
of the cartilaginous endplate.
Summary of Results: The infant cartilage endplate (CE) showed a highly
porous mineralized cartilage region that was connected to, but distinct from,
the underlying subchondral bone (SCB) and is an area of rounded pores that
likely contained chondrocytes. While the adult ovine tissue possessed similar
characteristics, the mineralized CE was significantly more compact and of
greater thickness and was thicker in the AF area than in the NP area. This
thinning is likely an adaptation to greater hydrostatic forces within the NP
region. In addition, the CE overlying the annulus contained tidemarks - a
histological feature that is typically associated with thickening of the calcified
region in articular cartilage and may indicate degenerative changes.
Conclusions: This work forms the basis for two separate studies of both
human and ovine spinal tissues to understand how the mineralized CE region
is altered with age in healthy subjects.
250
VALIDITY AND RELIABILITY OF THE JOINT COMMISSION
SUBJECT IDENTIFICATION CRITERIA FOR ASSESSING
THE QUALITY OF CHILDREN_S INPATIENT ASTHMA CARE
West S, Nkoy F, Fassl B, Stone B, Halbern S, Maloney C University of Utah,
Salt Lake City, UT.
Purpose of Study: The Joint Commission (JC) requires free standing children_s hospitals to report compliance with asthma quality core measures for
children hospitalized with acute asthma. Identification of children to be included in JC reporting is based on the primary ICD-9 discharge diagnosis
code. This approach excludes children with asthma not meeting this specific criterion. Objectives of this study are to 1) determine the sensitivity
and specificity of JC ascertainment of asthma discharges and 2) to describe
158
&
Volume 59, Number 1, January 2011
characteristics and outcomes of all asthma discharges stratified by JC ascertainment criteria.
Methods Used: This was a retrospective cross-sectional study of patients
2Y18 years admitted to a tertiary care children_s hospital between 1/08Y07/
08. Using administrative data, we identified all patients discharged with an
asthma ICD-9 diagnosis code (493.xx). We then classified patients as: True
Asthma (children whose symptoms and treatment were clinically consistent
with asthma based on chart review by 2 physicians), JC Criteria (primary
ICD-9 diagnosis of asthma), and Missed Asthma (children with true asthma
not identified by JC criteria). Administrative data was used to retrieve and
compare patient demographics and hospitalization outcomes.
Summary of Results: Of 476 patients with a 493.xx ICD-9 diagnosis code,
235 were classified as True Asthma. 184 of these matched JC Criteria, and
the remaining 51 were classified as Missed Asthma. Reliability of true
asthma ascertainment between the 2 physicians was 100%. JC criteria sensitivity was 78% and specificity 99%. Patient demographic characteristics and
readmission rates were similar among the groups.
However, patients with asthma not identified by JC criteria were more
likely to be admitted with concurrent respiratory infections (p G 0.001), had
significantly longer hospital stays (p= 0.002) and higher cost (p=0.001).
Conclusions: The JC criteria are moderately sensitive but highly specific at
identifying true asthma discharges. A quarter of patients with true asthma
were missed. Quality of asthma care may not be adequately represented by
utilizing JC identification criteria. Future studies should compare compliance
with CAC measures between asthma patients that are and are not identified
using JC criteria.
251
THE USE OF MULTIATTRIBUTE UTILITY THEORY IN THE
SELECTION OF RADIATION THERAPY TREATMENTS
Burt A1, Phillips MH2, Civan Hartzler A3 1University of Washington School
of Medicine, Seattle, WA; 2University of Washington Medical Center, Seattle,
WA and 3University of Washington Medical Center, Seattle, WA.
Purpose of Study: To determine the accuracy of utilities acquired from
Time Trade Off (TTO) and Standard Gamble (SG) exercises for the purpose
of individual decision-making and to evaluate the efficacy of multiattribute
utility algorithms (MAU) in modeling patient preference.
Methods Used: Ten health states, representing hypothetical treatment outcomes, were selected from the 48 total possible health states. Five residents
and physicians from the University of Washington Department of Radiation
Oncology completed direct health state and direct profile rankings based on
a visual analog scale. Utilities for the ten health states were elicited using
TTO and SG. TTO scores were adjusted using certainty equivalents; SG
scores were adjusted using prospect theory.
Summary of Results: TTO and SG were both able to accurately predict
direct health state ranking (TTO Q = 0.855, SG Q = 0.875) and direct health
profile ranking (TTO Q = 0.866, SG Q = 0.851). The MAU model based on
TTO utilities yielded utilities similar to adjusted TTO utilities acquired from
the interview (W = 14, p = 0.1836). The SG MAU model repeatedly underestimated corresponding interview utilities. Average error between the
MAU model and the acquired utility was greater for TTO than SG (TTO
average error = 0.096, SG average error = 0.059).
Conclusions: Both TTO or SG can effectively be employed to gather patient
utilities; utilities from either exercise can be used to construct a MAU model.
Adjustments to the MAU model to increase accuracy should be explored.
Utility gathering is a successful technique for increasing the patients_ role in
health care decision-making. Future work will employ these techniques to
elicit utilities from prostate cancer patients and subsequently utilized to rank
treatment plans.
252
PROGNOSTIC VALUE OF MAXIMAL COMPARED TO
3-MINUTE VASCULAR OCCLUSION TEST IN PATIENTS
WITH SEVERE SEPSIS OR SEPTIC SHOCK
Brisbane W3,4, Shen J3,4, Walters E1, Nguyen H1,2 1Loma Linda University,
Loma Linda, CA; 2Loma Linda University, Loma Linda, CA; 3Loma Linda
University, Loma Linda, CA and 4Loma Linda University, Loma Linda, CA.
Purpose of Study: Severe sepsis and septic shock involve immune activation and inflammatory response at the vascular endothelium leading to
microvascular injury. This diffuse endothelial disruption is responsible for
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
organ dysfunction, global tissue ischemia, and death. Measurement of tissue oxygen saturation (StO2) by near-infrared spectroscopy at the thenar
eminence during a 3-minute vascular occlusion at the forearm has been
shown to provide a marker of disease severity. In this study, we compared the
prognostic value of StO2 after the 3-minute vascular occlusion test (VOT) to
that of a maximal VOT.
Methods Used: A prospective observational study was performed in severe
sepsis or septic shock patients presenting to the ED and admitted to the
medical ICU. The lowest StO2 measurement (StO2min) at the thenar eminence (InSpectra 650 StO2, Hutchinson Technologies, MN) after a 3-min
VOT and maximal VOT was obtained at hour 0, 1, 2, 3, 4, 5, 6, 12, 24, 36, 48,
and 72. The 3-min VOT was performed with a tourniquet on the forearm
inflated for 3 minutes. The maximal VOT was similarly performed but with
the tourniquet inflated until StO2 reached a lowest steady state plateau.
Summary of Results: Sixteen patients were enrolled with age 65.9T
12.8 years and in-hospital mortality was 50%. APACHE II and lactate in
survivors vs. non-survivors were 18.75T8.1 vs. 28.25T10.2 (p=0.06), and
3.9T1.8 mmol/L vs. 8.3T3.6 mmol/L (p=0.01), respectively. Baseline StO2
and StO2min are illustrated in the Table.
Conclusions: StO2min after maximal vascular occlusion instead of only
three minutes of occlusion is better at identifying mortality in severe sepsis
and septic shock. The association of StO2min with oxygen transport mechanisms remains to be examined.
253
DOES EDUCATIONAL INTERVENTION CHANGE
PERCEPTION OF PREGNANT WOMEN TOWARDS CORD
BLOOD BANKING?
Shahani S1, Torres W2, Kirgan M2, Elsadat S3, Ezeanolue E1 1University of
Nevada School of Medicine, Las Vegas, NV; 2University of Nevada School
of Medicine, Las Vegas, NV and 3University of Nevada at Las Vegas, Las
Vegas, NV.
Purpose of Study: Potential indications for umbilical cord blood (UCB)
have increased over the last decade, yet the number of pregnant women who
elect to bank UCB has remained low. We sought to determine awareness and
perceptions toward Umbilical Cord Blood Banking (UCBB) among a cohort
of pregnant women and any change in attitude towards UCBB following an
educational intervention.
Methods Used: A cross-sectional survey of pregnant women 18 years and
older who attended prenatal care at a University affiliated clinic in Las Vegas
was conducted from February 1YMarch 1, 2010. Only participants who could
read English or Spanish were asked to complete a 25-question survey to assess
their awareness and knowledge about UCBB and to determine their willingness to donate UCB to a public or private bank. After completion of the survey, participants were randomly assigned to either of two groups. Group A
received an informational pamphlet from American College of Gynecologist
(ACOG). Group B did not receive the pamphlet. Both groups completed the
same questionnaire after 1 month. The University of Nevada Institutional
Review Board approved this study.
Summary of Results: 142 of 145 eligible participants completed the initial
survey for a response rate of 98%. 32% (45/142) of respondents were aware
of UCBB. 16% were willing to bank UCB, 52% of which to public banks.
The repeat survey response rate was 48% (68/142), with 47% of respondents
in Group A and 52% in Group B. Awareness was 59% in Group A and 42% in
Group B. Willingness to donate UCB was 47% in Group A, 73% of which to
public banks, and 34% in Group B, 42% of which to public banks. There was
no statistically significant difference in age, parity, ethnicity/race, educational
status, marital status, or income level between Groups A and B.
Conclusions: A significant proportion of pregnant women in our cohort
were unaware of UCBB and unwilling to donate blood for banking. Review
of available informational pamphlet led to an increased willingness to donate
cord blood among our cohort.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
254
TELEPHONE FOLLOW-UP OF EMERGENCY DEPARTMENT
PATIENTS: PATIENT CHARACTERISTICS AND CONTACT
PERCENTAGES
Oberfoell S1, Arora S2, Menchine M2 1DGSOM, Los Angeles, CA and
2
Keck School of Medicine at the University of Southern California, Los
Angeles, CA.
Purpose of Study: Emergency Physicians (EPs) fear being unable to
contact patients for follow-up or to provide results of important diagnostic
tests. Many EPs will not discharge patients until all diagnostic tests have
returned, even if the likelihood of a consequential finding is low. This
reluctance to discharge may result in longer wait times and Emergency
Department (ED) crowding. We aim to quantify the percentage of patients
who can be reached after leaving the ED. We hypothesize higher contact percentages among patients who are advised they will be contacted in the future
and who provide verified contact (intervention) than controls.
Methods Used: Based on our literature review, we estimated control follow
up percentages to be 75% and calculated that 626 total subjects would be
needed to achieve 80% power to detect a 10% absolute difference between
groups. We developed and field-tested a bilingual survey instrument on
ED patients. We conducted a randomized, controlled trial of a consecutive
sample of ED patients. Research assistants approached patients, obtained
informed consent, and enrolled subjects in the trial. Subjects were randomized to either control or intervention arms in blocks of 12. All subjects
completed a questionnaire that detailed the subject_s gender, race, ethnicity,
access to a telephone, and living conditions. In the control group, telephone
contact information was obtained from the registration intake. Subjects in
the intervention arm gave best and alternate contact information were explicitly
told of our intent to call them back. A maximum of 4 callback attempts were
made between 48 and 72 hours. The primary endpoint was successful telephone communication with patient or relative. Secondary endpoints are demographic factors associated with successful or unsuccessful callbacks.
Summary of Results: We enrolled a total of 309 patients (n=160 control,
n=149 intervention). We have contacted 75.6% of the control arm and 71.8%
of the intervention arm. This study is ongoing.
Conclusions: Halfway to our desired total enrollment we are not seeing
strong evidence that this intervention improves our ability to contact patients
by telephone. The contact percentages correspond with the predicted percentages from the literature.
Immunology and Rheumatology I
Concurrent Session
1:30 PM
Friday, January 28, 2011
255
REGULATION BY ESTROGEN AND PROGESTERONE
OF INTERFERON ALPHA SIGNALING IN
HUMAN LEUKOCYTES
Zheng J, Hughes G University of Washington School of Medicine,
Seattle, WA.
Purpose of Study: 90% of systemic lupus erythematosus (SLE) patients
are female, and the incidence peaks during reproductive years when estrogen
(Es) and progesterone (Pg) are at their highest. Studies show that Es increases
the risk of SLE in both humans and animal models, while Pg is protective.
Interferon alpha (IFN->) is a central pathogenic cytokine in SLE and can
directly activate multiple immune cell types to favor autoimmunity. Though
recent studies suggest a link between Es and IFN-> signaling in immune
cells, the relationship is not well understood; and very little is known about
the effects of Pg. To investigate the relationship between female reproductive
hormones and lupus autoimmunity, we tested the hypothesis that, in human
leukocytes, Es enhances IFN-> signaling while Pg suppresses it.
Methods Used: Peripheral blood mononuclear cells (PBMCs) were isolated
from blood of healthy donors. Cells were cultured in media, ethanol vehicle,
or physiologic concentrations of A-estradiol (Es), Pg, or a combination of both
hormones, with or without IFN->. Total RNA was isolated, and expression of
IFN-> inducible genes (CXCL10, MX1, PKR, IFIT1, ISG20) was measured by
quantitative PCR and normalized to housekeeping gene 18sRNA.
159
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Consistent with our hypothesis, we observed that Es
Conclusions: These results demonstrate that the tumor suppressor and
enhanced IFN-> signaling in human leukocytes; we did not observe an effect
of Pg. Es treatment significantly increased IFN-> induced expression of
CXCL10 (p=0.0273, two-tailed paired T-test). A similar trend was observed
for MX1, PKR and IFIT1. Es alone did not increase expression of any of
these genes. In addition, we observed that IFN-> signaling response to Es
was bi-modal, because PBMCs from some donors consistently showed Essensitivity while others did not.
Conclusions: Our research shows for the first time that Es can enhance
IFN-> induced gene expression in human leukocytes of healthy donors,
possibly by regulating IFN-> signaling pathways. This suggests a novel
mechanism by which Es increases a woman_s risk of developing SLE.
Additional experiments will allow us to determine whether Es regulates all
or a subset of IFN->-inducible genes and what factors determine Es sensitivity, such as Es receptor expression. We plan to extend these findings to
PBMCs of SLE patients to ask whether abnormal sensitivity or resistance to
Es or Pg may contribute to disease.
T-regulatory functions of Foxp3 are mediated through separate signaling
pathways. Our finding may have important application in the design of
autoimmune and cancer therapeutics targeting FOXP3.
256
EOSINOPHILIC VASCULITIS
Arena-Rook E1, Rankin D2, Colburn K1, Katsaros E1 1Loma Linda
University, Loma Linda, CA and 2Loma Linda University, Loma Linda, CA.
Case Report: Eosinophilic vasculitis is usually equated with Churg-Strauss
syndrome, a rare systemic, anti-neutrophil cytoplasmic antibody-associated
vasculitis. Churg-Strauss syndrome is most often found in patients with
asthma, but the onset can occur simultaneously with the diagnosis of asthma.
We present the case of a 20 year old male college wrestler, who initially
presented with a complaint of dyspnea while wrestling. The patient was found
to have recurrent pleural effusions for which he was referred to cardiothoracic
surgery. During the second decortication procedure, nodularity and thickening were discovered, and a wedge resection of the lung was performed.
The histology was notable for a reactive pleuritis and necrotizing granulomas
of the lung with eosinophilic necrosis, vasculitis and interstitial lymphoid
infiltrates. Rheumatology was consulted for suspected Churg-Strauss syndrome. Laboratory studies demonstrated a peripheral eosinophila of 10.2%.
However, serum anti-neutrophil cytoplasmic antibodies, human immunodeficiency virus antibodies, and antinuclear antibodies were all negative. The
initial fungal and parasitic stains were reported negative. A review of the
pathology slides led to the question of the patient_s participation in a popular
fad involving live crab ingestion, which he confirmed having done at a sushi
bar four months prior to onset of symptoms. Serum antibody tests confirmed
the diagnosis of Paragonimus westermani infection. P. westermani infections
are extremely rare in the United States, but found more commonly in endemic
areas of Asia. Pulmonary parasitic infections of P. westermani usually demonstrate necrotizing granulomas with an eosinophilic infiltrate; however,
there is little evidence in the literature of this parasitic infection leading to
a vasculitis. Our case illustrates a rare parasitic infection diagnosed as a
Paragonimus-induced eosinophilic vasculitis of the lung.
257
T-REGULATORY AND TUMOR SUPPRESSOR FUNCTIONS
OF FOXP3 ARE MEDIATED THROUGH SEPARATE
SIGNALING PATHWAYS
Heinze E1, Chan G2, Mory R1, Khavari R1, Nishimura R3, Weisbart R2
1
Olive View UCLA Medical Center, Sylmar, CA; 2Veterans Affairs Greater
Los Angeles Health Care System, Sepulveda, CA and 3University of
California, Los Angeles, CA.
Purpose of Study: The nuclear transcription factor, Foxp3, is required for
T-regulatory cell function (Tregs), but it also serves as a tumor suppressor
factor that inhibits development of breast cancer. Foxp3 requires NFAT
binding to maintain Treg function, but the requirement of NFAT for its tumor
suppressor activity is unknown. This study was designed to determine if
binding of NFAT to Foxp3 is required for its tumor suppressor activity.
Methods Used: Deletion mutants of Foxp3 were produced that prevent
binding of NFAT. Foxp3 and deletion mutants were transduced into cancer
cell lines as molecular fusion proteins with the protein transduction molecule,
mAb 3E10 Fv.
Summary of Results: Mutations that blocked NFAT binding to Foxp3 had
no effect on the cytotoxicity of FOXP3 in multiple cancer cell lines.
160
258
THE IMMUNOMODULATORY ROLE OF HIGH DOSE
INTRAVENOUS IMMUNOGLOBULIN IN A MURINE
MODEL OF BILIARY ATRESIA
Boguniewicz J1, Tucker R1, Brindley S1, Mack CL1,2 1University of
Colorado School of Medicine, Aurora, CO and 2University of Colorado
School of Medicine, Aurora, CO.
Purpose of Study: Biliary atresia (BA) is an inflammatory, sclerosing disease affecting the bile ducts of neonates causing choleostasis. While the incidence of BA is low, it is the most common indication for pediatric liver
transplantation. Current understanding of the pathophysiology of BA suggests that a viral insult initiates bile duct injury followed by progressive
autoimmune-mediated inflammation and fibro-obliteration of the bile ducts.
Intravenous immunoglobulin (IVIg) is composed of polyclonal, polyspecific
immunoglobulin and has demonstrated clinical benefit in several other autoimmune and inflammatory diseases. One mechanism of IVIg action is to increase Treg production, thereby decreasing autoimmune-mediated injury. The
purpose of our study is to determine if markers of bile duct injury are diminished and Treg production is increased in IVIg-treated BA mice.
Methods Used: Neonatal BALB/c mice were injected with rhesus rotavirus
(RRV) or Hanke_s Balanced Salt Solution (HBSS) 12Y18 hours after birth. On
day 7, 9, 11 BA mice were given 1g/kg high dose IVIg or albumin by intraperitoneal injection. Survival in each group was monitored and, at 14 days
post-injection, mice were sacrificed and blood, livers, and extrahepatic bile
ducts were collected. Direct bilirubin assay was performed on serum from
pools of 3Y4 mice. Liver immune cells were isolated by Percoll density gradient and quantification of liver Tregs was performed by flow cytometry.
Summary of Results: IVIg did not improve survival in BA mice compared
to albumin control. A significant decrease in direct bilirubin levels was observed in mice treated with IVIg (4.1T0.9 mg/dL) compared to albumin
(10.2T1.5) (unpaired t-test, p= 0.01) and untreated BA controls (10.5T1.9;
p=0.001). Increased Treg production was observed in IVIg-treated BA mice
(10.6T1.2% FoxP3) compared to albumin (7.5T0.1; p=0.06) and untreated
controls (5.9T1.4; p=0.07).
Conclusions: Immune therapy with high dose IVIg was associated with
decreased bilirubin suggesting diminished bile duct injury. An increase in
Tregs in IVIg-treated mice implies better control of autoimmunity and may be
a possible mechanism of action for IVIg in BA.
259
HUMAN TRANSITIONAL B CELLS IN NORMAL AND
AUTOIMMUNE PERIPHERAL BLOOD
Hsu S1, Weldon A2, Benitez A2, Baez I2, Colburn K1, Payne K2 1Loma
Linda University Medical Center, Loma Linda, CA and 2Loma Linda
University, Loma Linda, CA.
Purpose of Study: SLE and RA tissues have been shown to have higher
levels of autoreactive B cells suggesting a defect in the negative selection
process that occurs during the immature stages of B cell development. Mouse
studies indicate that immature stages of B cell development can be subdivided into the transitional stages T1 and T2. When these transitional B cells
are exposed to activation stimuli, the cells undergo receptor editing, anergy,
or apoptosis in order to eliminate autoreactive cells. The TNF family members APRIL and BAFF have been shown to promote survival of these transitional subsets in the mouse. Current studies have shown that SLE and RA
patients have elevated serum levels of BAFF and APRIL. High levels of
BAFF and APRIL potentially act on the transitional B cells promoting the
development of autoreactive B cells. For a more translational approach, we
utilized co-expression of CD21 and CD24 markers, which have been used
in mice to identify transitional and mature subsets. The aim of this study is to
identify transitional subsets in normal and autoimmune human peripheral
blood, and determine the effect of growth stimuli.
Methods Used: Human adult peripheral blood samples were stained for
seven-color flow cytometry to assess co-expression of CD24, CD21, IgD,
IgM, CD38, CD27, CD256 (APRIL), CD257 (BAFF), CD267 (TACI), and
BR3 (BAFF-R).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: The use of markers CD21 and CD24 allowed us to
identify T2, late transitional T2, and FM subsets in normal and autoimmune
(SLE and RA) peripheral blood. A lower percentage of T2 cells were identified in the SLE and RA patient samples compared to normal blood. In SLE
and RA samples, the receptors BAFF-R and TACI and the ligand APRIL
were elevated on CD19+, IgM+ B cells compared to normal peripheral blood.
Conclusions: The use of CD21 and CD24 markers can distinguish between
T1, T2, and FM B cell subsets. Fewer transitional subsets are present in
autoimmune peripheral blood and represent a more activated phenotype as
depicted by the higher expression of APRIL, BAFF-R, and TACI. The elevated levels of members of TNF family could contribute to the escape of
autoreactive B cells during the negative selection process during the transitional stages of B cell development.
260
AUTOIMMUNITY AND ASBESTOS RELATED DISEASE IN
AMPHIBOLE EXPOSED INDIVIDUALS OF LIBBY, MONTANA
Marchand LS1, Pfau J2 1University of Washington, Seatlle, WA and 2Idaho
State University, Pocatello, ID.
Purpose of Study: Autoimmune disorders stemming from an over-reactive
immune system are the cause of substantial disease and disability. Recent
literature has proposed a link between asbestos exposure and various autoimmune responses. Exposure to asbestos continues to occur throughout the
world making this issue a current health concern. Residents of the Libby,
Montana area have experienced significant exposure to amphibole asbestos
due to the mining of asbestos-contaminated vermiculite near the community.
This population exhibits an auto-immune-like disorder that has yet to be well
defined. Pleural and/or interstitial abnormalities are common findings in these
patients as both malignant and non-malignant disease types. This study investigated the relationship between autoimmunity, asbestos related disease
(ARD), and asbestos exposure.
Methods Used: Serum samples from Libby’s Center for Asbestos Related
Disease were evaluated through indirect fluorescence testing, checking for
anti-nuclear antibody (ANA) and cell based ELISA testing, looking for antimesothelial antibody (AMA).
Summary of Results: It was found that 62% of the Libby samples were
ANA positive, a frequency nearly double what would be expected of a healthy
population. On average it was determined that the odds of having pulmonary
disease was over 9 times greater for individuals with positive ANA tests
compared with individuals with negative ANA test results. The Libby population also exhibited a much higher level of AMA than did the control
group. A total of 23 (19%) Libby subjects were found to have significantly
high AMA levels and were established as AMA positive. Of these subjects
nearly 75% (17) had pleural disease. This meant that on average the odds of
having pleural disease is 4.72 times greater for individuals with positive
AMA levels compared to subjects without positive AMA. In contrast there
was no significant relationship between AMA positivity and interstitial disease. Sex and age were controlled for.
Conclusions: Data supports the possibility that AMA_s are contributing to
ARD. Additional research may be needed to further establish this relationship, but determining the nature of this association with clinical disease
progression might lead to an excellent target for therapeutic intervention and
disease screening.
261
LYMPHOCYTE PROFILES FROM THE SOFT TISSUES OF
METAL-ON-METAL HIPS
Tan T1,3, Sutthiruangwong P2, Al-Hamad M1, Magyar C2, Nelson S2,
Campbell P1 1Orthopaedic Hospital, Los Angeles, CA; 2SMOH/UCLA, Los
Angeles, CA and 3David Geffen School of Medicine at UCLA, Los Angeles, AR.
Purpose of Study: Aseptic loosening in metal-on-metal hips is classified
as being caused by wear debris or metal allergy. Metal sensitivity is associated with increased lymphocytic aggregates termed ALVAL compared to wear.
The purpose of this study was to characterize the immunological profiles of
lymphocytes and other immune cells in tissues around failed MM hips using
IHC to characterize the nature of the immune response.
Methods Used: Tissue samples from 21 archived MM hips were fixed in
formalin, paraffin embedded, sectioned and stained with H&E. The modes of
failure included 11 metal allergy, 8 high wear-related pseudotumor, and 2
other cases for controls. IHC was performed using a variety of histological
markers specific for immune cells. A 0-4 scoring system was used based on
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
the percentage of cells stained in a HPF. Digital image analysis was based on
recognition by color thresholding.
Summary of Results: 0-4 scoring of all cases showed more total T and T
cell subtypes (CD4,8,45RO) in metal allergy cases than high wear cases or
controls. Additionally, lymphocyte subpopulations in the metal allergy group
showed increased levels of both T and B cells. Plasma cells were less present
than T and B cells in metal allergy cases. Digital analysis produced values
incongruent with manual scoring. These differences occurred mainly in samples that contained wear debris, necrosis, blood, and heterogeneous tissue.
Conclusions: Originally explained as delayed type hypersensitivity (DTH),
soft tissue reactions containing a unique lymphocyte dominated histological
profile were revealed to contain T cells, B cells, and plasma cells. The presence
of B and plasma cells is inconsistent with DTH. Such heterogeneous lymphocytic populations, particularly in metal allergy, suggest an adaptive immune
response. More specifically, the prevalence of memory T cells and activated T
cells (CD45RO) suggest a T cell mediated adaptive response. An antibody
dependent (Type II) mechanism occurring alongside the DTH reaction may
explain the unexpected cells. Wear debris and constant metal ion exposure is
associated with a T cell lymphopenia and may alter lymphocyte profiles. Further
research requires elucidating the role of antibody processes in this response.
262
OUTCOMES AND COST-EFFECTIVENESS OF
RESIDENT-PERFORMED PALPATION-GUIDED
INTRAARTICULAR PROCEDURES IN A
TEACHING HOSPITAL
Yaqub S1, Ashraf U2, Norton H1, Gibb JI1, Sibbitt WL1, Bankhurst A1
1
University of New Mexico Health Sciences Center, Albuquerque, NM and
2
Texas Tech University Health Sciences Center, Amarillo, TX.
Purpose of Study: Patients are often apprehensive of undergoing an invasive procedure performed by inexperienced residents, even with attending
supervision. We hypothesized that intraarticular procedures would be less
painful and have better outcomes when performed by experienced attending
proceduralists compared to resident and fellows in training.
Methods Used: 326 joints underwent arthrocentesis followed by corticosteroid injection using the one-needle two-syring techniques performed by 1)
experienced proceduralists, 2) residents with direct attending supervision,
or 3) fellows with direct attending supervision. All procedures were performed with a mechanical syringe. Pain was measured with the 0Y10 cm
Visual Analogue Pain Scale (VAS) 1) prior to the procedure, 2) during needle
placement ( needle introduction, anesthesia, and arthrocentesis), 3) during
injection of corticosteroid, 4) 2 week post-procedure, and 5) 6 months postprocedure. A responder was defined as an asymptomatic joint at 2 weeks
(VASG 2 cm). Other outcomes included therapeutic duration (months), time
to next injection, cost/patient/per year, and cost/responder/year.
Summary of Results: As measured by the 10 cm VAS, procedural pain and
injection pain for residents and fellows were not statistically different than
attendings (p = 0.4 and 0.5, respectively). Similarly, pain at 2 weeks, pain at
6 months, percentage of responders, percentage of non-responders, therapeutic duration, time to next injection, cost/patient/year, and cost/responder/
year were not different between either residents or fellows compared to experienced proceduralists (p90.1 for all).
Conclusions: The procedural pain, outcomes, costs, and cost-effectiveness
of resident and fellow peformed intraarticular injection/aspiration procedures
performed with attending supervision in a teaching hospital are identical in
all measures of quality to those peformed by experienced proceduralists.
These outcome data are important to reassure patients and 3rd party payers
that quality of care at teaching hospitals is high.
263
CHOLESTASIS AS AN ATYPICAL AND MISLEADING
PRESENTATION OF KAWASAKI DISEASE: CASE REPORT
AND LITERATURE REVIEW
Brar CK1,2, Keosheyan R2,1, Arce-Hernandez E2 1UCSF Fresno Pediatrics,
Fresno, CA and 2Children’s Hospital Central California, Madera, CA.
Purpose of Study: Kawasaki Disease (KD) is an acute self-limited vasculitis of childhood that classically is characterized by fever, bilateral nonexudative conjunctivitis, erythema of the lips and oral mucosa, extremity
changes, rash, and cervical lymphadenopathy. Coronary artery aneurysms
or ectasia develop in 15% to 25% of untreated children. We report a case of
161
Western Regional Meeting Abstracts
Journal of Investigative Medicine
a 4 2 yr old with a very atypical presentation of KD involving cholestasis,
and review the literature on this rare presentation of KD.
Methods Used: Records from this patient_s 12-day hospitalization were
reviewed. A literature review was conducted using PubMed with key search
terms BKawasaki Disease[ and Bcholestasis[ or Bjaundice.[
Summary of Results: A previously healthy 4 2 yr old presented with fever,
emesis, abdominal pain, diffuse rash, and jaundice. Lab findings revealed
conjugated hyperbilirubinemia. She was initially diagnosed with cholangitis;
however, over the next few days the findings of classic KD progressively
developed. Her KD diagnosis and initiation of therapy occurred within the
optimal 10 day window from the onset of KD symptoms. After 2 doses of
intravenous immunoglobulin and pulse steroid therapy, she demonstrated
marked improvement. No coronary artery changes have been detected on
echocardiography.
Two case reports from the literature had similar presentations. The first
was a 6 yr old who had a delay in diagnosis due to his unusual presentation
and developed diffuse dilatation of his coronary arteries, which improved
with treatment. The second was a 10 yr old who also had a significant delay
in diagnosis and for whom treatment was initiated after the 10 day window.
He had a very complicated course consisting of multiple coronary artery aneurysms with severe multiple stenoses requiring surgical intervention.
Conclusions: Our case adds to the 2 similar reports published in the literature highlighting the importance of considering KD in a child who presents
with cholestasis, abdominal pain, and fever of unknown etiology. Not considering this atypical presentation may lead to a late diagnosis, and loss of the
10-day time window for early intervention, thus increasing the risk of coronary artery abnormalities.
264
ULTRASOUND-GUIDED ARTHROCENTESIS OF THE KNEE
USING NEW ASPIRATING SYRINGE TECHNOLOGIES
Kettwich LG, Sibbitt WL, Chavez-Chiang N, Delea S, Norton H, Bankhurst A
University of New Mexico Health Sciences Center, Albuquerque, NM.
Purpose of Study: Ultrasound image-guidance results in less traumatic,
more complete arthrocentesis. The present randomized controlled trial investigated new aspirating syringe technologies for image-guided arthrocentesis of the knee.
Methods Used: 42 ultrasound-guided arthrocentesis procedures of the knee
were randomized to new aspirating technologies: 1) a 60 ml vacuum syringe,
or 2) the 25 ml RPD (reciprocating procedure device) mechanical syringe.
The one-needle two-syringe technique was used. Completeness of arthrocentesis was determined by sonography. Outcome measures included patient
pain by the 10 cm Visual Analogue Pain Scale (VAS), synovial fluid volume,
complications, and therapeutic outcome at 2 weeks. Needle control of the
new technologies was measured in the linear displacement model and compared mechanical and vacuum syringes to conventional syringes.
Summary of Results: Both the mechanical syringe and the vacuum syringe
controlled the needle better than a conventional syringe, reducing unintended
forward penetration by 75% (3.6T0.5 mm) and 87% (12.0T4.2 mm), respectively (pG0.0001). Image-guided aspiration with both aspiration technologies permitted complete arthrocentesis with low levels of procedural
pain (10 cm VAS; Vacuum Syringe: 2.9T 3.1 cm, Mechanical Syringe: 3.1T
2.5 cm, p90.8) and significant fluid yield (Vacuum Syringe: VAS: 35T23 ml,
Mechanical Syringe: 34T27 ml, p90.9). The vacuum syringe permitted facile
automatic aspiration of up to 60 mls; the mechanical syringe permitted 25 ml
aspiration before a syringe exchange was required.
Conclusions: Ultrasound-guided arthrocentesis of the knee can be facilely
achieved with new aspirating syringe technologies with improved needle
control, enhanced patient safety, large volume aspiration, and complete joint
decompression.
265
OUTCOMES AND COST-EFFECTIVENESS OF
ULTRASOUND-GUIDED INVASIVE PROCEDURES
PERFORMED BY RESIDENTS IN A TEACHING HOSPITAL
Ashraf U1, Weis B1, Smalligan RD1, Gibb JI2, Norton H2, Sibbitt WL2,
Bankhurst A2 1Texas Tech University Health Sciences Center, Amarillo, TX
and 2University of New Mexico Health Sciences Center, Albuquerque, NM.
Purpose of Study: Ultrasound-guided procedures are becoming increasing
important in invasive medicine, but skills in ultrasound require considerable
162
&
Volume 59, Number 1, January 2011
experience. We hypothesized that ultrasound-guided procedures would be
less painful and have better outcomes when performed by experienced proceduralists compared to those performed by residents and fellows in training.
Methods Used: 256 joints underwent ultrasound-guided arthrocentesis followed by corticosteroid injection using the one-needle two-syringe technique
performed by 1) experienced proceduralists, 2) residents with attending supervision, or 3) fellows with attending supervision. All ultrasound-guided procedures were performed with a mechanical syringe. Pain was measured with
the 0Y10 cm Visual Analogue Pain Scale (VAS): 1) prior to the procedure, 2)
during needle placement (needle introduction, anesthesia, and arthrocentesis),
3) during injection of corticosteroid, 4) 2 week post-procedure, and 5) 6 months
post-procedure. A responder was defined as an asymptomatic joint at 2 weeks
(VASG 2 cm). Other outcomes included therapeutic duration (months), time to
next injection, cost/patient/per year, and cost/responder/year.
Summary of Results: As measured by the 10 cm VAS, procedural pain and
injection pain during ultrasound-guided procedures for residents and fellows
were not statistically different than experienced proceduralists (p = 0.6 and 0.5,
respectively). Similarly, pain at 2 weeks, pain at 6 months, percentage of responders, percentage of non-responders, therapeutic duration, time to next injection, cost/patient/year, and cost/responder/year were not different between
either residents or fellows compared to experienced proceduralists (p90.1 for all).
Conclusions: Ultrasound-guided procedures performed by trainees were
identical in all measures including procedural pain, outcomes, costs, and
cost-effectiveness to those performed by experienced ultrasound proceduralists. Since ultrasound guidance is becoming increasing important in invasive medicine, these equivalent outcome data are important to reassure
patients and 3rd party payors that quality of ultrasound procedures at teaching
hospitals is high.
Infectious Diseases
Concurrent Session
1:30 PM
Friday, January 28, 2011
266
IDENTIFICATION OF NEUTROPHILS AS A PREDOMINANT
SOURCE OF IL-1A PRODUCING CELLS IN THE EARLY
INNATE IMMUNE RESPONSE TO STAPHYLOCOCCUS
AUREUS CUTANEOUS INFECTION
Hebroni FF, Cho J, Ramos RI, Miller LS UCLA David Geffen School of
Medicine, Los Angeles, CA.
Purpose of Study: Staphylococcus aureus is a gram-positive bacterium responsible for a wide range of skin infections including superficial skin infections such as impetigo and more invasive skin infections such as cellulitis and
folliculitis. It has been previously demonstrated that the IL-1A/IL-1R pathway
is critical for neutrophil recruitment to a site of S. aureus infection in the skin.
However, the dynamics of IL-1A production during the course of infection and
the cellular sources of IL-1A have not yet been defined.
Methods Used: The goal of this study was to determine the kinetics of
IL-1A gene expression in real-time during a S. aureus cutaneous infection
by utilizing an IL-1A-dsRed reporter mouse line and to identify the IL-1A
producing cells using confocal microscopy.
Summary of Results: IL-1A gene induction could be detected in the
infected-skin lesions of live mice in real-time using in vivo fluorescence
imaging as soon as 8h post infection and peaked by 24h. The majority
of these early IL-1A-producing cells were detected with the neutrophil marker
7/4 whereas a significant but smaller number of IL-1A-producing cells were
detected with the monocyte/macrophage marker MOMA-2.
Conclusions: These findings identify what we believe to be a novel role for
neutrophils as the predominant source of IL-1A during the early innate immune response to cutaneous S. aureus infection.
267
INHIBITION OF HERPES SIMPLEX VIRUS (HSV)
GLYCOPROTEIN_S gB, gD, gH-gL INDUCED CELL FUSION
AND VIRAL SPREAD VIA HEPARAN MIMICS
Choudhary S1, Tiwari V1,2 1Western University of Health Sciences, College
of Osteopathic Medicine of Pacific, Pomona, CA and 2Western University of
Health Sciences, College of Optometry, Pomona, CA.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Purpose of Study: Multinucleated giant cells (polykaryocytes) resulting
from virus-induced cell fusion are a hallmark of herpes simplex virus-type-1
(HSV-1) infection. However, the role of HSV-1 3-O-sulfated heparan sulfate
(3-OS HS) receptor during cell fusion is poorly understood. The goal of this
study was to understand the role of heparan mimics in blocking the HSV- 1
infection.
Methods Used: HSV-1 glycoprotein induced quantitative reporter gene
(luciferase) based cell fusion assay was used. The counting of multinucleated
giant cells (polykaryocytes) was achieved by using fluorescent microscopy.
Effector Chinese hamster ovary (CHO-K1) cells expressing various combinations of HSV-1 glycoproteins were co-cultured with the 3-OST-3 expressing
primary cultures of human corneal fibroblasts (CF), a natural target cell-type
for HSV-1 infection. Pre-treatment of effector cells with heparinase enzyme
(4U/ml) and heparan mimics (5.6 nM) were used to demonstrate the role of
3-OS HS during cell fusion.
Summary of Results: We demonstrated that cell-to-cell fusion and polykaryocytes formation required expression of four essential HSV-1 glycoprotein
(gB, gD, gH-gL) and exhibited a very strong dependence on the expression of
heparan sulfate on CF. Further enzymatic removal of HS from CF surface by
heparinase-I treatment severely impaired the fusion reaction. Interestingly, the
incubation of effector cells expressing HSV-1 glycoprotein_s with heparan
sulfate mimics inhibited significantly both membrane fusion and polykaryocyte
formation.
Conclusions: Our results indicate that 3-OS HS could play a crucial role in
HSV-1 induced cell-to-cell fusion during corneal eye infection. Generation
of specific inhibitors targeting against HSV-1 gD epitope that interacts with
3-OS HS would be beneficial in the development of therapeutics to prevent
viral spread in the corneal stroma. Here, we provide evidence for the physiological significance of 3-OS HS mediated cell-to-cell fusion.
268
THE ROLE OF FC-GAMMA RECEPTOR IIIB VARIATION IN
CONDITIONING INTERFERON-GAMMA LEVELS IN
SUB-SAHARAN AFRICAN CHILDREN WITH SEVERE
MALARIAL ANEMIA
Chaudhury A, Perkins D University of New Mexico, Albuquerque, NM.
Purpose of Study: Plasmodium falciparum infections are associated with
the development of severe malarial anemia (SMA), one of the leading causes
of childhood morbidity and mortality in sub-Saharan Africa. Although, the
genetic and molecular basis of SMA is only partially defined, severity of
malarial disease is affected by dysregulation of host-derived inflammatory
mediators. Fc-F Receptor IIIB (FcFRIIIB) facilitates binding to the Fc portion of IgG1 and IgG3, which are important in modulating immune responses
and in protection against P. falciparum in endemic areas. The granulocyte
antigens NA1 and NA2 are the two recognized allelic forms of FcFRIIIB. The
overall aim of this study was to determine the association of FcFRIIIB variants with severe malarial anemia and cytokine production in children presenting at a rural hospital in western Kenya with P. falciaprum parasitemia.
Methods Used: Polymerase chain reaction and Multiplex Assay were utilized to determine how variation in FcFRIIIB conditions functional changes
in cytokine production in children (n=383) with P. falciparum malaria.
Summary of Results: Stratification of children according to parasitemic
status revealed that the SMA group [hemoglobin (Hb) G6.0 g/dL] had lower
levels of circulating IFN-F than the non-SMA group (HbQ6.0 g/dL) (P=0.11).
Multivariate analyses controlling for the confounding effects of age, gender,
HIV-1 status, bacteremia, and sickle-cell trait showed that the heterozygous
FcFRIIIB NA1/NA2 genotype were protected from SMA (OR=0.758, [95%
CI: 0.40-1.20]) relative to homozygous NA1 or NA2. Functional analyses
showed that heterozygosity at FcFRIIIB (NA1/NA2) was associated with
significantly increased circulating IFN-F levels relative to homozygous
individuals (NA1; P=0.02).
Conclusions: Finding important genetic determinants of phenotypic outcomes in children with malaria, would enable clinicians to predict which
children are at a higher risk for developing SMA, and may promote earlier
treatment interventions to reduce mortality. Results from this study illustrate
the need for further exploration to determine if suppressed levels of IFN-F
secretion due to variation in the FcFRIIIB receptor could be ameliorated by
exogenous administration of IFN-F to reduce severe malaria in individuals
with P. falciparum infection.
* 2011 The American Federation for Medical Research
269
PROINFLAMMATORY IMMUNE RESPONSE AND
PUERPERAL GROUP A STREPTOCOCCAL SEPSIS
Spargen PF1, Augustine NH1,3, Clark EA2, Davis SM2, Martins TB3,
Wilson AR3, Silver RM2, Hill HR1,3 1University of Utah, Salt Lake City, UT;
2
University of Utah, Salt Lake City, UT and 3ARUP Institute for Clinical and
Experimental Pathology, Salt Lake City, UT.
Purpose of Study: Group A streptococcus (GAS) is a serious and potentially fatal, source of postpartum infection. Bacterial virulence factors influence
pathogenesis but do not effectively predict disease susceptibility or severity.
The objective of this study was to determine if altered maternal innate immune
response is associated with puerperal GAS sepsis.
Methods Used: Case-control analysis of innate immune response in women
with a history of puerperal GAS sepsis. Cases had febrile illness and/or
endometritis during the postpartum period and at least one positive culture.
Cases were further stratified into mild vs. severe; severe cases had either 1)
disease requiring surgical exploration/debridement, 2) admission to ICU, 3)
toxic shock syndrome, or 4) hospitalization 9 14 days. Cases were at least
one year post-infection at the time of analysis. Controls were healthy subjects
matched 1:1 by age, parity, and race. Immunoassays (Luminex) assessed cytokine/inflammatory marker production in tissue culture supernatant obtained
from stimulated peripheral blood mononuclear cells. Concentrations of 13
cytokines/inflammatory markers (IFNF, IL1A, IL2, IL4, IL5, IL6, IL8, IL10,
IL12, IL13, TNF>, IL2r, and soluble CD40L) were measured in response to
heat-killed emm1 and emm28 GAS, the M-serotypes most closely associated
with puerperal morbidity.
Summary of Results: Ten controls, 10 severe cases, and 6 mild cases were
analyzed. In response to stimulation, CD40L, IL1A, IL2r, and IL6 production
were significantly increased in both severe and mild cases compared to
controls (CD40L mean response 131.7 vs. 69.2 pg/ml, p=0.0001; IL-1A
mean response 1143.3 vs. 502.3 pg/ml, p=0.0001; IL2r mean response 13.7
vs. 6.4 pg/ml, p=0.001; IL6 mean response 4841.1 vs. 2534.0 pg/ml p=0.01).
Conclusions: Puerperal GAS sepsis is associated with altered maternal
innate immune responses. The markedly increased production of inflammatory cytokines may contribute to susceptibility and severity of puerperal
GAS disease.
270
THE INCIDENCE OF BLOOD AND BODY FLUID EXPOSURE
IN OUR TRAINEES
Ives CM1, Inaba K1, Barmparas G1, Swadron S2, Teixeira P1, Saadi Z1,
Lenzini M1, Demetriades D1 1LAC+USC Medical Center, Los Angeles, CA
and 2LAC+USC Medical Center, Los Angeles, CA.
Purpose of Study: The incidence of blood and body fluids exposure in
trainees is not well documented. The purpose of this study was to document
exposure rates in medical students and high risk specialty residents.
Methods Used: Third and fourth year medical students (MS), Surgery and
Emergency Medicine residents (RS) were anonymously surveyed regarding
percutaneous, cutaneous and mucous membrane exposures in the 6 months
preceding survey administration.
Summary of Results: 770 surveys were completed. 37.7% (58/154) of RS
and 15.4% (95/616) of MS were exposed. The incidence of exposure increased in a stepwise fashion with year of training. 31.2% involved a trauma
patient. Exposure occurred most frequently while performing a procedure
(80.6%) followed by observation of a procedure (5.7%) and blood draw
(2.9%). The most frequent RS exposure was solid needlestick (31.3%), followed by hollow needlestick (20.9%) and splash in eyes (13.9%); for MS,
splash in eyes (28.4%), was followed by solid needlestick (27.5%) and splash
on skin (18.3%). Contributing factors were uncontrolled fluid splash (44.3%),
patient movement (12.5%), fatigue (10.8%), unguarded sharps (9.7%) and
unsafe instrument pass (6.8%). Only 27.3% of exposures were reported to
occupational health. 41.5% of source patients underwent HIV and Hepatitis
testing. Overall, 35.4% of incidents reported to occupational health services
triggered the administration of prophylaxis. None reported known seroconversion related to the exposure.
Conclusions: Body fluid exposure is common in trainees, increasing with
level of training to greater than 25% in final year medical students and 70% in
final year residents. Less than a third were reported to occupational health.
163
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Identification of the reasons for non-reporting and potential target areas for
the prevention of exposure is warranted.
271
EFFECTIVENESS OF ROUTINE METHICILLIN RESISTANT
STAPHYLOCOCCUS AUREUS SCREENING
&
Volume 59, Number 1, January 2011
Conclusions: Our study demonstrates that patients undergoing UCBT are
at high risk for invasive fungal infections. Additional studies to elucidate risk
factors for IFI and future prospective trials examining antifungal prophylactic
strategies in UCBT may help provide new options for prevention.
273
BOPA IS REQUIRED FOR PERSISTENCE OF BRUCELLA
Jack I1, Ezeanolue E1,2, Elsadat S1, Rae Tuano K1 1University of Nevada
School of Medicine, Las Vegas, NV and 2University of Nevada Las Vegas,
Las Vegas, NV.
Purpose of Study: Hospitalized patients are routinely screened for MRSA
as a means to reduce hospital acquired MRSA. We sought to determine how
reliable screening tests identify patients with proven MRSA infection.
Methods Used: We retrospectively reviewed charts of all patients aged
0Y18 years hospitalized in a community based children_s hospital between
November 2008YNovember 2009. Patients with culture proven invasive
MRSA infection were identified and their screening tests for MRSA were reviewed to determine any correlation. We assumed a null hypothesis that culture proven invasive MRSA results were independent of screening results and
an alternative hypothesis that there was an association. Data analysis was performed by Fisher_s exact test using SPSS (18) with a p-value G 0.05. Our study
was approved by the Institutional Review Boards of University of Nevada, Reno
and Sunrise Hospital.
Summary of Results: 3453 patients were screened for MRSA during our
study period. 3.45% (119/3453) had a positive test result. 150 patients were
identified to have culture proven MRSA infection. Among those with culture
proven MRSA infection, 42.1% (32/76) had a positive screening test result:
nasal cultures accounted for 6.25% of positive cultures, 28.1% throat cultures, 12.5% urine cultures, 37.5% rectal, 6.25% axillary and 9.38% other sites.
The null hypothesis was rejected in support of the alternative hypothesis.
Conclusions: A significant proportion of patients with invasive MRSA infection also had a positive MRSA screening test result. Routine screening for
MRSA potentially could reduce Hospital acquired MRSA infection by indentifying colonized patient who can then be isolated.
Atluri V, Tsolis R UC Davis, Davis, CA.
Purpose of Study: Brucella species cause brucellosis, a chronic disease
characterized by undulating fever, in about 500,000 people every year. Survival of bacteria in macrophages and chronic persistence in the reticuloendothelial system (RES) of the host, two key features of Brucella pathogenesis,
are dependent on the VirB type IV secretion system (T4SS). Previous studies
have shown that the T4SS is involved in both trafficking of the Brucellacontaining vacuole to an endoplasmic reticulum-derived niche and activation
of genes involved in inflammation and immunity. However, the identity of the
T4SS-secreted molecules contributing to these two phenotypes is unknown.
A recent screen for genes co-regulated with the T4SS structural genes
identified BopA as a protein with features of a potential effector. Genomic
analysis revealed a second protein, designated BopB, which is similar to
BopA at the amino acid level.
Methods Used: To determine whether BopA and/or BopB play a role in
intracellular survival and persistence mutants lacking one or both of these genes
($bopA, $bopB, and $bopAbopB) were constructed and characterized.
Summary of Results: The $bopA mutant was attenuated for survival
in immortalized murine macrophages and exhibited reduced persistence
in mice. In addition, T4SS dependent inflammasome activation, required
for production of the cytokine IL-1 beta and IL-18 is partially dependent
on BopA. The $bopB and $bopAbopB mutants are currently being
characterized.
Conclusions: These results suggest that BopA contributes to survival of B.
melitensis in macrophages, possibly by interacting with host cell proteins
after cytosolic translocation.
272
INCIDENCE OF INVASIVE FUNGAL INFECTIONS IN
UMBILICAL CORD BLOOD TRANSPLANT RECIPIENTS
274
MACROLIDE RESISTANT TREPONEMA PALLIDUM:
ASSOCIATION WITH MOLECULAR STRAIN TYPE
Chow V1,2, Milano F1, Delaney C1,3, Xie H1, Pergam SA1,2 1Fred Hutchinson
Cancer Research Center, Seattle, WA; 2University of Washington, Seattle, WA
and 3Seattle Children’s, Seattle, WA.
Purpose of Study: Infections are a common cause of morbidity and mortality following umbilical cord blood transplantation (UCBT), however limited
data on invasive fungal infections (IFI) exist in this population. The focus of
this study was to better understand the incidence and potential risk factors for
IFI in UCBT recipients.
Methods Used: This retrospective cohort study evaluated all patients who
underwent UCBT at the Fred Hutchinson Cancer Research Center (FHCRC)
in Seattle, WA between 2006 and 2009. Data were collected using medical
records and the FHCRC_s Long-Term Follow-Up database. Only proven or
probable IFI as defined by international criteria were included in analyses.
Incidence rates were determined by standard methods and 95% confidence
intervals (CI) were estimated based on a Poisson distribution. Risk factors for
developing IFI were assessed using chi squared or Wilcoxon rank-sum
analyses where applicable.
Summary of Results: There were 103 patients eligible for inclusion in the
cohort. Median age of the cohort was 30.4 years (interquartile range [IQR]
14, 48), and most underwent myeloablative conditioning (71/103 [69%]) and
received a double cord transplant (88/103 [85%]). A total of 20 cases of IFI
(5 proven, 15 probable) were found in 17 patients (16.5%). The median onset
of first IFI was 62 days post-transplant (IQR 27, 94). The cohort specific
incidence rate of IFI during the first year post-transplant was 0.97 cases per
1000 patient days (95% CI 0.59, 1.5). Invasive Aspergillus comprised the
majority of cases (13/20 [65%]), while other IFI included Zygomycetes (3),
Scopulariopsis (1) and Fusarium (1); only 2 (2%) developed candidal infections during follow-up. A total of 4/17 (24%) patients died as a direct result
of their IFI. When comparing UCBT recipients who did and did not develop
IFI, graft-versus-host disease, time to engraftment, age, conditioning regimen,
or CMV serostatus were not associated with the development of IFI.
Grimes MD1, Godornes C1, Tantalo L2, Marra CM2, Lukehart SA1 1Univ. of
Washington School of Medicine, Seattle, WA and 2Univ. of Washington
School of Medicine, Seattle, WA.
Purpose of Study: Syphilis is usually treated with intramuscular benzathine penicillin G, which is painful and can be problematic due to penicillin
sensitivity. Azithromycin can be administered orally, has proved effective in
treatment of early syphilis, and has been considered as a possible alternative
drug. Treatment failure of azithromycin and other macrolides, however, has
been documented in some patients and is linked to point mutations, termed
A2058G and A2059G, in the Treponema pallidum (T. pallidum) 23s rRNA
genes. The purpose of this study was to determine the prevalence of these
mutations in T. pallidum samples obtained in Seattle during the past ten years,
and to correlate these mutations with molecular strain type.
Methods Used: Subjects with syphilis were enrolled in an ongoing study of
neurosyphilis; most subjects were HIV-infected. Genomic DNA was isolated
from two sources: 1) T. pallidum strains isolated by inoculating rabbits with
patient_s blood, or 2) directly from patient blood. Strain typing had been
previously conducted on the samples based on analysis of three DNA target
regions. Samples were screened for the A2058G and A2059G mutations
using nested PCR amplification and RFLP analysis.
Summary of Results: The percentage of samples carrying macrolide resistance mutations increased from 21.4% in 2002 to 100% in 2009. The
A2058G mutation was more common and was present from 2002Y2009,
while the A2059G mutation was observed only from 2002Y2005. Each mutation was detected in multiple strain types, but the strain types harboring the
two mutations were distinct.
Conclusions: T. pallidum macrolide resistance mutations in Seattle are associated with specific molecular strain types, and the use of strain typing can
aid in definition of sexual networks. We speculate that selection for, or development of, resistant strains of T. pallidum may be facilitated by the use of
azithromycin prophylaxis in HIV-infected persons. Further investigation of
164
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
specific regional prevalence of macrolide resistance mutations in T. pallidum
is required before informed local decisions can be made concerning the use of
azithromycin as an alternative to penicillin in the treatment of syphilis.
Morphogenesis and Malformations
Concurrent Session
1:00 PM
Friday, January 28, 2011
275
VIABILITY WITH GERMLINE HRAS G12V MUTATION IN
COSTELLO SYNDROME IS DUE TO REDUCED MRNA
EXPRESSION ASSOCIATED WITH DINUCLEOTIDE
SUBSTITUTION
Stevenson DA1, Carey J1, Calhoun A1, Yu P2, Vaughn C2, Dobrowolski S2,
Swensen J2 1University of Utah, Salt Lake City, UT and 2Salt Lake City, UT.
Purpose of Study: Costello syndrome is most commonly due to p.G12S
activating mutations in HRAS with relatively weak transforming activity.
G12V has the highest transforming activity and greatest frequency in human
cancers, but rare in Costello syndrome. G12V has been hypothesized to result
in early lethality and a more severe phenotype in most cases: mice with this
mutation have high perinatal mortality. We report the molecular and phenotypic findings of a 12-year-old boy with an attenuated Costello phenotype
with a germline p.G12V mutation resulting from a dinucleotide substitution,
c.35_36delinsTG without evidence of mosaicism.
Methods Used: DNA and RNA extracted from peripheral blood; DNA also
extracted from hair, cheek swab, and urine. Bidirectional sequencing for the
coding regions and intron/exon boundaries of HRAS performed on DNA
from blood and mutation confirmed in other samples. Random-primed cDNA
prepared from RNA was used for analysis of HRAS expression.
Summary of Results: The boy had hypertrophic cardiomyopathy, mild
ptosis, posteriorly angulated ears, pectus deformity, reflux, normal growth
parameters, and mild developmental delay but full scale IQ of 76. No history
of malignancy, ulnar deviation, or abnormalities of hair. HRAS sequencing
identified a c.35_36delinsTG (p.G12V) mutation in all tissues sampled
without evidence of mosaicism. A significant reduction in the level of the
mutant allele was observed in cDNA.
Conclusions: Attenuated phenotypes have been reported with specific
mutations in Costello syndrome but not with p.G12V. Although our patient
had hypertrophic cardiomyopathy, the remaining phenotype was comparatively attenuated. This is even more surprising given the association of the
p.G12V mutant with higher stage and lethality in carcinomas. We demonstrate reduced levels of the mutant allele in cDNA and speculate this reduction results from altered mRNA processing. The 2 other individuals with
Costello syndrome previously reported with a p.G12V mutation also had
dinucleotide substitutions. We propose that the unusual dinucleotide substitutions result in viability due to reduced expression of the mutant alleles.
Western Regional Meeting Abstracts
Summary of Results: The RCS-LTC cells produced mRNA for all three
enzymes, most prominently for P3H2, in contrast with normal adult rat cartilage and SAOS2 cells which lacked mRNA expression for P3H2 and P3H3
respectively. Quantitative comparison of P3H mRNA expression between
the RCS-LTC cells and rat cartilage yielded similar expression of P3H1, a
È2-fold increase in P3H2 and a È0.5-fold increase in P3H3 expression in
the RCS-LTC cells. Mass-spectometric analysis of the RCS-LTC collagen
showed near complete 3Hyp formation at Pro944, a secondary site previously
found occupied in the type II collagen of vitreous, but not cartilage.
Conclusions: The high relative abundance of P3H2 mRNA expression
coupled with high occupancy of Pro944 in the RCS-LTC matrix implicates
P3H2 in the formation of 3Hyp at Pro944. Taken together with previous findings, it is possible that 3Hyp formation at Pro944 may be a signaling mechanism for the prevention of cleavage of the collagen N-propeptide, thereby
maintaining the long, thin fibrils as seen in RCS-LTC cells and vitreous.
Funded under NCRR Grant TL1 RR 025016 and NIAMS Grant AR
036794.
277
PRENATAL DIAGNOSIS OF AMYOPLASIA CONGENITA
Muller E1, Enns G1, Hintz S2, Barth R3, Hudgins L1 1Stanford University,
Stanford, CA; 2Stanford University, Stanford, CA and 3Stanford University,
Stanford, CA.
Case Report: Amyoplasia congenita, the most common form of arthrogryposis multiplex congenita, is a sporadic disorder, as classically defined by
Hall in 1983. It is characterized by symmetric limb involvement with fatty
and fibrous tissue replacement of hypoplastic muscle, vascular facial skin
lesions, and normal cognitive development. It is a static condition, and there
is accumulated evidence that it can result from a variety of prenatal vascular
insults to the ventral horn cells of the developing fetal spine during critical
periods, as first suggested by Reid in 1986 (discussed in Gaitanis et al, 2010).
Here we present the first published case diagnosed by fetal MRI. Fetal ultrasound at 19 weeks of gestation revealed flexion deformities of the upper
and lower extremities and an absence of fetal limb movement. Chromosome
analysis and FISH for common aneuploidies via amniocentesis were normal.
Fetal MRI at 22 weeks of gestation confirmed ultrasonography findings, and
revealed increased subcutaneous signal indicative of edema or increased fat
in the muscle, and suggestive of muscle atrophy, in all four limbs. Gestation
and delivery proceeded without complication. The patient was born at full
term and noted to have a weak cry, normal gag reflex, and the typical physical
features of amyoplasia congenita were identified. No pterygia were present.
Postnatal brain and spine MRI revealed no specific anomalies of the CNS.
The infant was discharged home on the second day of life.
Amyoplasia congenita is a distinct form of arthrogryposis with a good
prognosis overall. We suggest that the characteristic fatty replacement and
muscular atrophy in the limbs may be visualized on fetal MRI, and in the
context of features consistent with arthrogryposis may allow prenatal diagnosis of this condition.
References:
276
COLLAGEN SUBSTRATE SPECIFICITY OF PROLYL
3-HYDROXYLASES
Farnand AW1,2, Weis ME2, Kim LS2, Fernandes RJ2, Eyre DR2 1University
of Washington, Seattle, WA and 2University of Washington, Seattle, WA.
Purpose of Study: Current attention has focused on prolyl 3-hydroxylases
(P3Hs), a class of enzymes responsible for 3-hydroxylation of certain prolyl
residues in the collagen triple helix, forming 3-hydroxyproline (3Hyp). Mutations in the genes encoding P3H1 and associated proteins of its enzyme
complex were found to cause recessive forms of severe osteogenesis imperfecta (OI). The goal of this study was to investigate the expression levels of the
three P3H enzymesVP3H1, P3H2 and P3H3Vin a cell line known to form
3Hyp at several sites in type II collagen, to better understand their different
substrate specificities.
Methods Used: Using qPCR, expression of the three P3H enzyme genes
was assayed in the rat chondrosarcoma cell line, RCS-LTC, and in normal
adult rat cartilage. Similarly, the expression profiles of these genes, and other
genes related to the functionality of the P3Hs, were compared qualitatively by
RT-PCR and were compared to expression levels in the human osteosarcoma
cell line, SAOS-2. Collagen produced by the RCS-LTC cells was analyzed for
3Hyp formation at known sites by mass-spectometry.
* 2011 The American Federation for Medical Research
Gaitanis JN, et al. Pediatr Neurol 2010;43:142Y147.
Hall JG, et al. Am J Med Genet 1983; 15:571Y590.
Reid COMV, et al. Am J Med Genet 1986; 24:701Y710.
278
MITOCHONDRIAL PATHOLOGY IN ANGELMAN
SYNDROME
Niemi A1, Cox R1, Scharfe C2, Cherry A3, Enns G1 1Stanford University,
Stanford, CA; 2Stanford University, Stanford, CA and 3Stanford University,
Stanford, CA.
Purpose of Study: Mitochondrial diseases share many clinical features
with Angelman syndrome including developmental delay, seizures, gait
ataxia, microcephaly, and GI symptoms. AS is caused by the loss of the
maternally imprinted copy in the 15q11.2-q13 Angelman/Prader-Willi syndrome (AS/PWS) region. Approximately 70% of patients with AS have a
cytogenetically detectable deletion in this region. We report a patient with AS
who had mitochondrial pathology on muscle biopsy to further the understanding of pathophysiology of AS.
Methods Used: Microarray 105k CGH. Review of electronic chart &
medical literature.
165
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Our patient is an 8-year old male with develop-
Purpose of Study: The 19p13.13 microdeletion syndrome (M. Dolan, et al.
mental delay, microcephaly, seizures, ataxia, and constipation. Extensive
biochemical evaluation, sequencing of multiple mitochondrial genes,
and mitochondrial respiratory chain analysis were normal. After a muscle
biopsy showed mitochondrial hyperplasia and ragged red fibers suggesting
mitochondrial dysfunction, he was referred to our center for evaluation
of possible mitochondrial disorder. Urine organic acids showed dicarboxylic
acids and lactate, findings often seen in patients with mitochondrial disorders.
An array CGH revealed a 4.85 Mb deletion from 15q11.2 to 15q13.1
encompassing the UBE3A gene, consistent with a diagnosis of AS.
Conclusions: Evidence of mitochondrial pathology on a muscle biopsy
in a patient with AS suggests that UBE3A may regulate mitochondrial
biogenesis and/or function. UBE3A is an ubiquitin ligase and ubiquination
is important in maintaining the structural and functional integrity of
the mitochondrion. Brains of mice with AS in which the maternal UBE3A
allele is mutated have abnormal mitochondria that exhibit partial defect in
mitochondrial respiratory chain (Su et al. 2009). We are also sequencing 524
nuclear mitochondrial genes in this patient as part of research study to
determine if we can identify any potential modifying genes that may contribute to mitochondrial pathology in AS. Further studies are needed to
determine whether some of the manifestations of AS may be explained by
secondary mitochondrial pathology.
Genetics in Medicine 2010, 12(8):503Y511) is based on a series of cases
with macrocephaly and overgrowth with 19p13.13 microdeletions of varying sizes, the smallest area of overlap being 311Y340 kb and encompassing
16 genes. We present a patient with this phenotype and a smaller deletion
within this same region. This narrows the critical region and suggests NF1X
as a candidate gene responsible for this overgrowth syndrome.
Methods Used: A whole genome 1,800,000 single nucleotide polymorphism
(SNP) (Affymetrix, Inc.) chromosome microarray (LabCorp) was used on DNA
derived from blood.
Summary of Results: An 8 year old male with autism, tall stature, macrocephaly and moderate-severe mental retardation presented for genetic assessment. He had pectus excavatum, constipation, and an undescended testis.
The head circumference was 59 cm (9998th %ile) and height 140 cm (94th
%ile). His eyelids were puffy, eyebrows were straight and his face was triangular. Palpebral fissures were downslanting. Skin on the palms was dry and
loose. He was tactile defensive with stereotypic movements and poor eye
contact. He spoke in 3 word phrases. SNP microarray identified a 188 kb
deletion of 19p13.13 including NF1X (nuclear factor-1-X), a candidate gene
in the original report. Maternal analysis with a region specific BAC FISH
probe was normal. Paternal analysis is pending. NF1X is a transcription
factor expressed in the developing human brain and skeleton. Mutations
in NF1X have recently been reported in patients with Sotos-like and
Marshall-Smith syndromes (V. Malan, et al. AJHG 2010 Aug 13;87(2):
189Y98). Our patient’s clinical characteristics will be presented and compared with other patients with NF1X deletions and mutations from the
literature.
Conclusions: This report refines the clinical and molecular characteristics
of the 19p13.13 microdeletion syndrome and narrows the critical region to a
188 kb area that includes the NF1X gene. Mutations in NF1X have been
recently reported in patients with other overgrowth syndromes. We conclude
that the overgrowth macrocephaly phenotype associated with microdeletions of 19p13.13 is caused by loss of the NF1X gene.
279
RECURRENT ACUTE PANCREATITIS, DISTINCTIVE
FACIAL APPEARANCE, AND 3-HYDROXYISOBUTYRIC
ACIDURIA: A PREVIOUSLY UNRECOGNIZED
PRESENTATION OF PEARSON SYNDROME
Calhoun AR1, Botto L1, Pohl J1, O’Gorman M1, Pasquale M2,1, Longo N1,2
1
University of Utah, Salt Lake City, UT and 2University of Utah, Salt Lake
City, UT.
Purpose of Study: Pearson syndrome is the most severe of the mitochondrial
DNA deletion syndromes and typically presents in infancy with transfusiondependent anemia and variable pancreatic insufficiency. We report a patient with
a novel presentation characterized by recurrent acute pancreatitis, distinctive
facial appearance, and abnormal biochemical findings.
Methods Used: Retrospective chart review.
Summary of Results: The patient presented at 13 months of age with
emesis, lethargy, poor oral intake, and dehydration, associated with failure to
thrive and mild enlargement of the liver. Cognition was normal. Distinctive findings included a round face with full cheeks, and acute pancreatitis
(abdominal pain, elevated lipase/amylase). Abdominal imaging revealed a
pancreatic cyst (2 cm diameter) and mild liver enlargement. Brain MRI and
MR spectroscopy were normal. At presentation she had anemia, mild leucopenia and thrombocytopenia, mild metabolic acidosis and normal liver
enzymes. Over the following months she was re-admitted eight times for
recurrent acute pancreatitis (lipase up to 2,908). She had chronically elevated
lactate (10T3.8 mM (n=56), normal 0.7Y2.1 mM), elevated alanine, low
arginine, lactic and pyruvic aciduria, and persistently elevated excretion of
3-hydroxy-isobutyric and 2-ethyl-3-hydroxypropionic acids suggestive of
mitochondrial dysfunction. Renal tubulopathy was also present, with aminoaciduria, glycosuria and tubular acidosis. She progressed to severe intestinal dysmotility (even with pancreatic enzyme supplements), liver
failure, and transfusion-dependent pancytopenia. Bone marrow biopsy
showed trilineage hematopoesis with moderate dyserythropoesis, vacuolization of erythroid and myeloid precursors without ringed sideroblasts.
Mitochondrial DNA testing identified a large, heteroplasmic deletion consistent mtDNA deletion syndrome.
Conclusions: Pearson syndrome should be suspected in children with recurrent pancreatitis without a known cause, especially in the setting of
multiorgan system involvement and suggestive biochemical testing.
280
A NEWLY DESCRIBED OVERGROWTH, MACROCEPHALY,
DEVELOPMENTAL DISABILITY SYNDROME IS DUE TO A
MICRODELETION OF 19P13.13 AND LOSS OF THE
NF1X GENE
Clark RD1, Papenhausen PR2 1Loma Linda University School of Medicine,
Loma Linda, CA and 2Laboratory Corporation of America, Research
Triangle Park, NC.
166
281
NEWBORN WITH CHOROIDAL FISSURE CYST AND
PANHYPOPITUITARISM
Chitkara R1, Rajani A1, Bernstein J2, Hudgins L2, Shah S3, Hahn J4, Hintz S1
1
Stanford University, Palo Alto, CA; 2Stanford University, Palo Alto, CA;
3
Stanford University, Palo Alto, CA and 4Stanford University, Palo Alto, CA.
Case Report: Little is known about choroidal fissure cysts and the resultant
spectrum of medical complications. We describe the case of a 37-week male
infant with known prenatal fetal findings of a large choroidal fissure cyst,
bilateral clubfeet and screen positive for trisomy 18 and Smith-Lemli-Opitz
(SLO) Syndrome. Fetal MRI at 31 weeks showed a large right-sided choroidal fissure cyst (28mm) and asymmetric ventricles. Amniocentesis was
declined. Delivery was unremarkable. Physical exam was notable for a fixed
and dilated right pupil, bilateral clubfeet and micropenis. The patient was
admitted to the NICU for further work-up including Genetics, Neurology,
Neurosurgery, Ophthalmology, Endocrine and Orthopedic consultations.
Brain MRI re-demonstrated a large right-sided choroidal fissure cyst with
mass effect and midline shift. He underwent cyst fenestration and cystoperitoneal shunt placement on day of life 7. Chromosomal analysis showed 46
XY and 7-deoxycholesterol level was normal, ruling out SLO. Comparative
genomic hybridization showed a 3.64Mb interstitial deletion involving most
of band 10q26.12 and the proximal half of band 10q26.13; the deletion was
confirmed by FISH analysis. Parental samples were negative for the deletion.
Ophthalmologic exam showed anomalous optic nerves, pupil asymmetry and
right cranial nerve III palsy. Endocrinologic workup was notable for low
thyroid, cortisol and testosterone levels for which he was started on levothyroxine, hydrocortisone and testosterone, respectively. Due to severe panhypopituitarism, brittle diabetes insipidus requiring DDAVP, episodes of
sepsis, feeding problems and failure to thrive requiring G-tube, the patient
remained in the hospital for 3 months. He has since been admitted for shunt
revision, Achilles tendon release and fever. This case represents the severe
end of the spectrum of medical complications for children with choroidal
fissure cysts. It is the first to our knowledge that describes the associated
finding of deletion of part of chromosome 10. It highlights the importance of
meticulous evaluation and management and follow-up of these patients, and
for more research in this area.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
282
LONG TERM FOLLOW-UP OF A 61-YEAR-OLD WOMAN
WITH WOLF-HIRSCHHORN SYNDROME DUE TO AN
UMBALANCED INHERITED 4P;13Q TRANSLOCATION
Leon EL, South S, Carey J University of Utah, Salt Lake City, UT.
Purpose of Study: Clinical description and delineation of the chromosomal alteration in the oldest known patient with Wolf-Hirschhorn syndrome
(WHS). To advance knowledge of natural history.
Methods Used: Clinical observation and phenotype analysis, chromosome
and CGH microarray analyses.
Summary of Results: The proposita was born to healthy nonconsanguineous parents, a 23-year-old G1P091 mother and 26-year-old father, of
Caucasian descent. Family history is unremarkable. She was born postterm, via
induced vaginal delivery, following an uncomplicated pregnancy. She was
small for gestational age, with BW of 2.350 kg, and her length was 46 cm.
Her neonatal course was remarkable for poor sucking. Her developmental
milestones proceeded slowly, and by 18-months-old she was able to sit up with
assistance. She has never been able to walk, feed herself or speak. Seizures
started at about 11 months of age; first with grand mal convulsions and later
in her childhood with minor motor seizures. She was diagnosed with agenesis
of corpus callosum at 18 months. She now has intermittent episodes of seizures
and she has been off seizure medication since she was 36. At her initial evaluation (26 years) her growth parameters were below the 3rd centile. Her facial
features were typical for WHS. She had bilateral fifth finger clinodactyly,
scoliosis, spasticity in four limbs, and decreased muscle bulk. Conventional
G-banded cytogenetic studies detected a deletion in 4p. Her mother had a
balanced translocation between 4p and 13q. CGH microarray analysis in
order to characterize the size of the deletion/duplication is pending.
Conclusions: This is the oldest reported patient with WHS; she has an
inherited unbalanced translocation. Previous studies indicate that approximately 15Y45% of WHS cases are due to inherited translocations. The 4p/13q
translocation is a rare translocation causing WHS.
283
NEUROLOGICAL FINDINGS IN TWO SIBLINGS WITH
TAYBI-LINDER SYNDROME
Pierce MJ1, Morse R2, Curry C1 1UCSF Fresno, Madera, CA and
2
Dartmouth-Hitchcock Medical Center, Lebanon, NH.
Case Report: In 1967 Taybi and Linder described two siblings, from consanguineous parents, with dwarfism, skeletal dysplasia, and brain malformations. The syndrome is now known as microcephalic osteodysplastic
primordial dwarfism type I/III, or Taybi-Linder syndrome, and is characterized by microcephaly, growth retardation, and characteristic skeletal findings
including: platyspondyly, short iliac wings, flat/irregular acetabular roofs, and
short and flat long bones.
We present the neurologic and morphologic profiles of two siblings, female
and male, born to non-consanguineous, Caucasian parents. The emphasis of
the report is on the neurological profile of the disease, which includes brain
malformations, intractable epilepsy, sensory deficits, profound developmental
delay, and neuroendocrine dysfunction. We also present novel correlative
neuroimaging (MRI) and electroencephalographic (EEG) findings.
Both siblings were born prematurely, with growth parameters at birth
below the 25th centile for weight and length, and head circumference less
than 10th centile. Facial dysmorphias included hypertelorism, down-slanting
palpebral fissures, micrognathia, and glabellar nevus flammeus. Early skeletal radiography in both was consistent with Taybi-Linder syndrome.
Karyotypes were normal and a targeted microarray in the sister was also
normal. She had significant congenital heart disease, including VSD, PFO,
and PDA. Her brother had micropenis with hypospadias and bilateral
cryptorchidism.
Both sibs had profound cognitive impairment, with blindness secondary
to optic nerve hypoplasia and sensorineural hearing loss secondary to bilateral Mondini malformations. Neonatal onset seizures appeared in both, refractory to anticonvulsants and only partially responsive to ACTH. EEG_s
demonstrated diffuse encephalopathy. Neuroendocrine abnormalities included
central hypothyroidism, pseudo-hyperaldosteronism, and diabetes insipidus.
MRIs of both siblings were notable for anomalies in cortical gyration, cerebellar hypoplasia, and hypoplasia of the corpus callosum.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
The sister expired at age four years from complications of a viral illness.
The brother is now 33 months and is tracheotomy and G-tube dependent, with
limited development.
284
HEME OXYGENASE-1 DEFICIENCY IMPAIRS PLACENTAL
VASCULAR FORMATION AND EMBRYONIC DEVELOPMENT
Zhao H, Azuma J, Kalish FS, Wong RJ, Stevenson DK Stanford University
School of Medicine, Stanford, CA.
Purpose of Study: Placental vasculature formation is critical for establishing the fetomaternal interface and the development of healthy offspring.
Heme oxygenase-1 (HO-1), the enzyme in heme degradation, plays a role in
angiogenesis/vasculogenesis and is highly expressed in the placenta. Its deficiency is associated with several pregnancy disorders. Our objective was to
study the effects of HO-1 deficiency on placental vasculature formation and
embryonic development.
Methods Used: Placentas from crossbred HO-1 heterozygote (Het) mice
were harvested at various gestational ages and genotyped by PCR. Changes
in placental morphology were detected by H&E staining. Uterine NK (uNK)
cells, involved in early decidual vasculature development, were identified by
DAB staining. Maternal vasculature was visualized from casted placentas and
imaged by microCT. Expressions of decidual factors, including PlGF, VEGF,
Flt1, Flk1, TGFA, IFNF, IL10, iNOS and eNOS, were quantified by RT-PCR.
All comparisons were made between age-matched Het and wild-type (WT)
placentas.
Summary of Results: Compared to WT, Het matings overall yielded
smaller fetuses and placentas. In Het placentas, spiral arteries were less dilated and junction zones were thinner by H&E staining. 3D images revealed
that Het placentas had thinner (2.1 vs 2.4mm) maternal vascular regions, less
total vessel volume (8.3 vs 9.0mm3) and reduced vessel volume in the labyrinth (3.1 vs 4.4mm3). Het placentas also had more spiral artery branching,
but with smaller diameters (0.30T0.05 vs. 0.41T0.04mm). Expression of both
PlGF (placental growth factor) and VEGF (vascular endothelial growth
factor), the two key vascular regulators, was significantly elevated in Het
deciduas (up to 2.7-fold and 10-fold, respectively). uNK cells in Het deciduas
appeared significantly smaller (21.8T4.7 vs. 26.3T3.9mm) and with more
granularity.
Conclusions: HO-1 deficiency results in abnormal spiral artery dilation and
remodeling, and poor labyrinth vessel development. The defects are likely
due to irregular expression of vascular growth factors and improper structure
of uNK cells. Therefore we speculate that HO-1 plays an important role in
placental vasculature development and HO-1 deficiency may contribute to
pregnancy complications, such as preeclampsia, spontaneous abortions and
premature births.
285
PRENATAL EXPOSURE TO MATERNAL CIGARETTE
SMOKING HAS A GREATER EFFECT ON FETAL GROWTH
THAN NICOTINE ALONE
Malleske DT, Fung C, Campbell M, Kaur R, Yu X, Callaway C, McKnight
R, Albertine K, Lane RH University of Utah, Salt Lake City, UT.
Purpose of Study: We hypothesized that prenatal exposure to maternal
cigarette smoking, but not nicotine alone, will cause IUGR in the offspring.
Methods Used: Pregnant Sprague-Dawley rats were exposed to MCS or
nicotine alone (NA). MCS exposure (10 cigarettes per dam) occurred daily
from day 11 (d11) of gestation until term (d21), excluding weekend days. NA
dams received intraperitoneal osmotic pumps containing either a nicotine
solution (1, 3 or 6 mg nicotine/kg/d) or saline on d13. All pups were delivered
surgically and weighed on d21. Dams were weighed on d11 (MCS) or d13
(NA) and d21. Serum cotinine levels from dams were determined at time of
delivery via ELISA.
Summary of Results: MCS pups averaged 21% smaller than unexposed
pups (PG0.01), with males and females affected equally. Plasma cotinine
levels in MCS dams were 235+/j43ng/ml (human reference range, 200Y
800ng/ml for active, daily smokers). Dams exposed to NA with similar
plasma cotinine levels (243+/j80ng/ml) produced pups less growth restricted
(5% males, 7% females; PG0.05) than MCS pups. Average initial weight of
dams, litter sizes and placental weights at delivery were not different between
control, MCS and NA animals. Average weight of MCS dams at delivery was
13% lower than controls (PG0.05).
167
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Conclusions: These data suggest that prenatal MCS exposure has a greater
effect on fetal growth and maternal weight gain than nicotine alone.
*DTM and CF contributed equally to this work.
286
MATERNAL TOBACCO SMOKE EXPOSURE AND
INTRAUTERINE GROWTH RESTRICTION ALTER HEPATIC
SREBP1A AND -2 LEVELS IN A GENDER SPECIFIC MANNER
Zinkhan E, Wang Y, Hale M, Jiang C, Kaur R, Yu X, Callaway C, Fitzhugh M,
Brown A, Campbell M, Fung C, Malleske DT, McKnight R, Joss-Moore L,
Albertine K, Lane RH University of Utah, Salt Lake City, UT.
Purpose of Study: Maternal tobacco smoke exposure (MTSE) and uteroplacental insufficiency (UPI) cause intrauterine growth restriction (IUGR).
Both MTSE induced IUGR (MTSE-IUGR) and UPI induced IUGR (UPIIUGR) increase the risk for hypercholesterolemia in adult humans and rats.
The major detrimental component of cholesterol is low density lipoprotein
(LDL). Increased LDL production leads to significant adult morbidity and
mortality. Morbidity and mortality from elevated LDL is more prevalent in
males. Serum and hepatic LDL levels are regulated by sterol-responsive
element binding protein 1a (SREBP1a) and -2 (SREBP2). Despite the importance of SREBP1a and -2 in the regulation of LDL throughout life, it
is unknown whether MTSE-IUGR and UPI-IUGR alter SREBP1a and -2
mRNA and protein levels. We hypothesized that MTSE-IUGR and UPIIUGR decreases SREBP1a and -2 mRNA and protein levels in males at birth
with more substantial decreases seen in the MTSE-IUGR model.
Methods Used: To test this hypothesis we used both a MTSE-IUGR and a
UPI-IUGR rat model to determine mRNA levels of SREBP1a and -2, and
protein levels of SREBP2 at birth in both genders. MTSE-IUGR was achieved
by cigarette smoke exposure from day 11 of pregnancy to term. UPI-IUGR was
achieved by bilateral uterine artery ligation at day 19 of pregnancy.
Summary of Results: MTSE-IUGR decreased SREBP2 mRNA levels to
81% +/j 5% (p=0.058) in males, with no change in SREBP1a in either
gender. UPI-IUGR decreased male SREBP1a mRNA to 64% +/j 8%
(pG0.05), SREBP2 mRNA to 60% +/j 10% (p=0.06), and SREBP2 protein
to 29% +/j 5% (pG0.01), with no change in females.
Conclusions: We conclude that MTSE-IUGR and UPI-IUGR decrease
SREBP1a and -2 in a gender and model specific manner. Model specific
alterations in SREBP1a mRNA levels suggest separate mechanisms through
which each insult impacts gene expression. These mechanisms may include
vasoconstriction from nicotine, hypoxia, or toxins from smoke exposure.
We speculate that MTSE-IUGR and UPI-IUGR hepatic lipid profiles will
demonstrate increased intrahepatic cholesterol in IUGR males.
287
DISTINCT PHENOTYPE ASSOCIATED WITH INTERSTITIAL
LONG ARM DELETION OF CHROMOSOME 4q13.3Y q21.1
Golabi M1, Hall B2, Cotter P3 1Sutter Pacific Medical Center, San Francisco,
CA; 2University of Kentucky, Lexington, KY and 3Children’s Hospital of
Oakland, Oakland, CA.
Purpose of Study: Report a new case of 4q13.3Yq21.1, document two
previously reported cases with identical features, and define the unique features associated with this deletion.
Methods Used: Oligo nucleotide array CGH analysis was performed on our
proband’s DNA. This test was comprised of 99,000+ DNA probes. The
DECIPHER database documented clinical descriptions of two previously
recorded cases with overlapping deletions.
Summary of Results: The proband, a female, was born at 37 weeks gestation to a 35-year-old Mother. CVS showed a 46,XX karyotype. At birth the
proband presented with significant IUGR and microcephaly. She was noted to
have dysmorphic facial features, such as a bulbous nasal tip with wide alae,
abnormal helices, cleft palate, and small mandible. She had a short neck,
broad chest, rhizomelic shortening of upper and lower extremities, short
broad thumbs, distal digital fat pads of fingers and toes, and a plantar lipoma
on her left heel. She had truncal hypotonia, delayed milestones, and sensorineural hearing loss. Tests showed a normal brain MRI, delayed bone-age,
and elevated IGF 1 on two occasions.
Previous reports of patients with similar deletions: (1) A male with prenatal
growth retardation and microcephaly with a deletion of 4q13.3Yq21.21. (2)
A female with prenatal onset of short stature, abnormal ears/hearing deficit, strabismus, abnormal palate, and delayed tooth age with a deletion of
168
&
Volume 59, Number 1, January 2011
4q13.2Yq21.21. Although the proband_s deletion was smaller, she has a distinctive phenotype characterized by microcephaly, cleft palate, and sensorineural hearing loss. She also had a plantar lipoma, which is a rare finding. We
suggest that a 4q deletion should be added to the list of conditions associated
with plantar lipomas. Also of interest, she has significant short stature and
delayed bone age despite a two-fold increase in IGF1 levels. Further evaluation
will determine whether the proband might also have a mutation within the IGF1
receptor or whether one of the genes with the 4q deletion is involved in IGF1
regulation.
Conclusions: We propose the microdeletion of 4q13.3Yq21.1 is associated
with a distinct and recognizable phenotype.
Neonatology Y General II
Concurrent Session
1:15 PM
Friday, January 28, 2011
288
BATH-RELATED BURN IN THE NEONATE
Rayjada N, Sardesai S, Cayabyab RG Division of Neonatal Medicine, Keck
School of Medicine,University of Southern California, Los Angeles, CA.
Case Report: We report a case of a 9-day old full term female infant with
bath-related burns involving 70% total body surface area, managed with
Meplex and Acticoat dressings in neonatal intensive care unit (NICU).
Introduction: Neonatal burns are rare and can be fatal. There is limited literature on the management of significant burns in this unique patient population.
Case Report: A 9-day old full term female neonate with bath-related burns
was transferred to our Burn Unit and later to the NICU from an outside
Emergency Room after nine hours of initial injury. She had sustained 70%
total body surface area burns (7Y10% third-degree burns and 60% seconddegree burns) affecting the arms, legs, abdomen, back and maxillary region
of the face. Early and aggressive fluid resuscitation was initiated. Upon arrival to the Burn Unit the wounds were covered with Meplex and Acticoat
dressings. More severe burns were also kept moist using red-robin catheters
to deliver saline underneath the dressings. The dressings allowed for nearly
painless wound management. Multiple courses of antibiotics and anti-fungals
were initiated secondary to blood cultures that grew Pseudomonas Aeruginosa
and Candida Albicans. The patient was discharged after 80 days. Of note, at
discharge the burns were well-healed with erythematous areas of skin and
fibrotic changes noted on the chest, abdomen, back, lower extremities, buttocks
and labial region. No surgical intervention or skin grafts were necessary. The
patient was recently seen at the Burn Clinic. She was noted to have healed
very well without scarring and had full range of motion of all extremities.
Hypertrophic and erythematous areas were noted around the inner thighs
and mon pubis. She will continue to be followed as an outpatient with the
Burn Clinic.
Discussion: Because of the unique physiology in the neonatal period the
management of thermal injuries in this population poses many challenges. The
goals of our treatment included patient stabilization, preservation of cosmetic
appearance, joint functionality and pain management. Acticoat is a suitable
dressing for neonates who have sustained burn injury, with the advantage of
minimal handling as the dressing need only be changed every 3Y7 days.
289
BOLUS VERSUS LONGER INFUSION OF ENTERAL
FEEDINGS FOR PRETERM NEONATES
Corbin LL, Smith S OHSU, Portland, OR.
Purpose of Study: Developmental immaturity of the gastrointestinal tract
precludes immediate full enteral feedings for many preterm infants. Previous
studies suggest that preterm infants who receive slow infusion of enteral
feedings have improved maturation of intestinal motility when compared to
preterm neonates fed by rapid bolus. Our hypothesis was that infusion of
gavage feedings over 120 minutes will lead to a 33% reduction in feeding
intolerance, as defined by the proportion of total feedings withheld, when
compared to bolus gavage feedings given over no more than 30 minutes.
Methods Used: This study is a single center, randomized controlled trial
comparing two methods of enteral feeding for preterm neonates less than
32 weeks postmenstrual age and birth weight between 500 and 1500 grams.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Group 1 infants (Bbolus[) received intermittent bolus feedings given over
no more than 30 minutes. Group 2 infants (Bdrip[) received intermittent
slow infusion feedings over 120 minutes. The primary outcome was the
proportion of total feedings withheld for feeding intolerance. The proportion
of feedings withheld was analyzed using generalized estimating equations
(GEE) to account for the clustered nature of having multiple days of data
for each child. The model utilized a logit link with exchangeable correlation
structure and, in addition to treatment (drip vs bolus), investigated the effects
of gestational age on the outcome. Informed parental consent was obtained
for all participants and the study was approved by the OHSU Institutional
Review Board.
Summary of Results: A total of 29 infants were recruited for the study,
15 bolus and 14 drip. Odds of a feed not being given were 67.8% lower (95%
CI: 21.8Y86.7 percent lower) for infants on drip feeds compared to the bolus
feed (Wald X^2(1)=6.3, p=0.012). After controlling for gestational age
(23Y27 wks vs 28Y30 wks), the odds of withholding a feed remain 68%
(95% CI: 26Y86%) lower for those on drip feeds compared to those on bolus
feeds (Wald X^2(1)=7.08, p=0.008).
Conclusions: Our study showed that infants receiving feeds over 2 hours
had less feeding intolerance, as measured by the number of feeds withheld,
than those infants who received bolus feedings. Further analysis will determine the number of days to achieve full feeds and to regain birth weight in
this population.
290
INCREASED AMINO ACIDS POTENTIATE GLUCOSE
STIMULATED INSULIN SECRETION IN THE OVINE FETUS
Gadhia M, Maliszewski A, O’Meara M, Thorn SR, Lavezzi JR, Hay WW,
Brown LD, Rozance PJ University of Colorado, Denver, CO.
Purpose of Study: Previous studies of intrauterine growth restricted fetuses demonstrate reduced amino acid supply to the fetus. These fetuses are
also characterized by decreased insulin secretion. It is unknown whether
chronically increased fetal amino acid supply would stimulate insulin secretion, even in normally growing fetuses. We hypothesized that chronically
increasing the fetal amino acid supply would increase glucose and arginine
stimulated insulin secretion (GSIS, ASIS) in the ovine fetus.
Methods Used: Singleton ovine fetuses at 113Y120 days gestation
(term=148 days) were given an intravenous infusion of a complete mixture of
amino acids (AA group, n=8) or saline (CON group, n=8) for 10Y14 days.
Fetal branch chain amino acids (BCAA), glucose, insulin, pH, blood gasses,
and hematocrit were measured during the infusion. On the final day, fetal
GSIS and ASIS were measured using a square wave hyperglycemic clamp
and arginine bolus. Pancreatic insulin content was measured with ELISA.
Summary of Results: Fetal BCAA concentrations increased 50% in the
AA group compared to CON (PG0.05). Glucose decreased over time in the
AA group (22.7T1.5 baseline vs. 17.9T0.4 mg/dl final day, PG0.01) but not
in CON. pO2, O2 saturation, and O2 content tended to decrease during
days 5Y10 in the AA group and then returned to baseline. Hematocrit, pH,
and insulin concentrations were not different between the groups during the
infusion period. However, there was a pronounced increase in early phase
insulin concentrations (15 min: 2.06T0.53 AA vs. 0.80T0.20 ng/ml CON,
PG0.01) and this difference was sustained during the hyperglycemic clamp in
the AA group (105 min: 1.38T0.23 AA vs. 0.70T0.14 ng/ml CON, PG0.001).
ASIS appeared increased in the AA group but this was not statistically significant. Pancreatic insulin content was not different between groups.
Conclusions: Chronically increased amino acid supply to the fetus potentiated insulin secretion in response to glucose. Because GSIS was potentiated
more than ASIS, we speculate that amino acids upregulate glucose metabolism and generation of secondary messengers in the beta-cell.
291
INCREASED VITAMIN E INTAKE IS ASSOCIATED WITH
HIGHER >-TOCOPHEROL CONCENTRATION IN THE
MATERNAL CIRCULATION, BUT HIGHER
>-CARBOXYETHYL HYDROXYCHROMANS
CONCENTRATION IN THE FETAL CIRCULATION
Go M1, Didenco S1, Gillingham M1, Leonard S2, Traber M2, McEvoy C1
1
Oregon Health and Sciences University, Portland, OR and 2Oregona State
University, Corvallis, OR.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Purpose of Study: Vitamin E may prevent oxidative stresses implicated
in perinatal disease processes, but studies suggest fetal vitamin E stores remain low despite maternal supplementation. We hypothesize that the placenta limits vitamin E transfer by increasing it_s metabolism, or the fetal liver
actively metabolizes the vitamin, which would be reflected by the concentrations
of vitamin E metabolites, carboxyethyl hydroxychromans (>- and F-CEHC), in
the fetal circulation. We measured >- and F-CEHC levels in maternal and
umbilical cord blood pairs and examined their relationships to circulating
vitamin E (>- and F-tocopherol) and maternal dietary vitamin E intake.
Methods Used: At least one fasting blood sample and a previous day_s
24-hr diet recall were done during pregnancy in healthy, non-smoking
women. All samples were analyzed for >- and F-tocopherol, >- and F-CEHC
and total lipid concentrations (n=17 pairs, all full term).
Summary of Results: Umbilical cord blood and maternal concentrations
of >- CEHC (30.2 T 28.9 vs. 50.4+52 nmol/L, P = 0.07) and F-CEHC
concentrations (104.5 T 61.3 vs. 141.7+78.5 nmol/L; P = 0.08) were not
significantly different, but the metabolite to tocopherol ratios were significantly higher in cord blood than in maternal blood (>-CEHC:>-tocopherol =
5.7+5.5 vs. 1.6+1.9; P G 0.01 and F-CEHC:F-tocopherol = 389.6+291.5 vs.
86.8+64.4;, P G 0.001). Maternal >-tocopherol:total lipids ratio were correlated with cord blood >-CEHC (r = 0.67, P G 0.003, and higher maternal
vitamin E intakes were associated with the higher cord blood >-CEHC concentrations (r = 0.75, P G 0.003).
Conclusions: This is the first report of CEHC concentrations in cord blood.
Although the results show similar CEHC concentrations in maternal and
cord blood samples, the >-CEHC:>-tocopherol ratios in the fetal circulation
were triple those in the maternal circulation. The higher intake of vitamin E
during pregnancy resulted in elevated fetal metabolite levels, without increasing the fetal blood vitamin E concentration. Studies are needed to further define vitamin E metabolism in pregnancy and the function of vitamin E
metabolites.
292
PROLONGED HYPOGLYCEMIA DECREASES INSULIN
SECRETION, NOT A-CELL POPULATION IN OVINE FETUSES
Lavezzi JR, O’Meara M, Thorn SR, Brown LD, Hay WW, Rozance PJ
University of Colorado School of Medicine, Aurora, CO.
Purpose of Study: Intrauterine growth restriction due to placental insufficiency (PI-IUGR) is characterized by fetal hypoglycemia, hypoinsulinemia,
hypoxia, decreased amino acids, increased catecholamines, nearly absent
glucose stimulated insulin secretion (GSIS), and specific reduction of the
A-cell population. We hypothesized that prolonged fetal hypoglycemia over
the last 40% of gestation, independent of placental insufficiency, would
replicate these defects.
Methods Used: Control fetuses (C; n=5) were compared to fetuses whose
mothers received an 8 wk insulin infusion reducing maternal and fetal glucose 40% (HG; n=5). Four other HG fetuses (HG+I) received a direct insulin
infusion with concurrent dextrose infusion to prevent fall in glucose for the
final wk. Glucose, insulin, amino acids, O2, norepinephrine, GSIS, arginine
stimulated insulin secretion (ASIS), pancreatic weight and fetal weight were
measured at the end of the infusions. A-cell area was measured as the proportion of pancreatic sections which stained insulin+. A-cell mass is the
product of A-cell area and pancreatic weight.
Summary of Results: Glucose (22.8T1.7 C vs. 10.6T0.4 HG mg/dl;
PG0.0001), and insulin (0.47T0.06 C vs. 0.18T0.03 HG ng/ml; PG0.005) were
lower in HG fetuses. O2 and norepinephrine were similar. Taurine was increased (PG0.01), glutamate decreased (PG0.001), but all other amino acids
were unchanged in HG fetuses. GSIS was present but decreased 45% in HG
fetuses (PG0.05) and ASIS was preserved. A-cell area did not change
(4.57T0.52 C vs. 4.66T1.21 HG %). The reduction in A-cell mass (32%) in
HG fetuses was proportional to the decrease in pancreatic (37%) and fetal
(40%) weights. By design in HG+I fetuses, insulin increased (0.89T0.19 ng/ml,
PG0.005) and glucose did not change (9.2T0.8 mg/dl). GSIS was absent and
ASIS decreased (PG0.0005). A-cell area, A-cell mass, fetal, and pancreatic
weight were not statistically different from HG fetuses.
Conclusions: Prolonged hypoglycemia decreased GSIS, though not as
strikingly as seen in PI-IUGR. Similarly, the A-cell population was not specifically decreased relative to fetal and pancreatic weight. These results
contrast those of severe human IUGR and animal models of PI-IUGR and
define chronic experimental hypoglycemia as a model of IUGR with a unique
A-cell phenotype.
169
Western Regional Meeting Abstracts
Journal of Investigative Medicine
293
EFFECTS OF A DIRECT FETAL AMINO ACID INFUSION
ON OXYGENATION AND ACID-BASE BALANCE IN
FETAL SHEEP
Maliszewski A, Gadhia M, O’Meara M, Thorn SR, Hay WW, Rozance PJ,
Brown L University of Colorado, Denver, CO.
Purpose of Study: Human maternal high protein supplementation during
pregnancy increased the risk for small for gestational age birth. Maternal
amino acid infusion in pregnant sheep results in competitive inhibition of
amino acid transport across the placenta, increased fetal oxygen (O2) consumption, fetal hypoxia and acidosis. A direct fetal amino acid infusion, bypassing competitive inhibition of amino acid transport across the placenta,
will increase fetal O2 consumption but preserve acid-base balance.
Methods Used: Singleton fetal sheep were intravenously infused with a
complete amino acid mixture (AA, n=8) or saline (C, n=10) for an average of
12 days during late gestation. A mixed model ANOVA was performed to
determine the effects of treatment group (AA or C) and day of infusion on
fetal arterial branched chain amino acid (BCAA) concentrations; pH; blood
gasses; hematocrit; plasma lactate, glucose, and insulin concentrations, and
glucose/O2 quotient. On the final day of infusion, a metabolic study was
performed to determine rates of umbilical blood flow, fetal O2 consumption,
and fetal glucose and lactate uptakes.
Summary of Results: Fetal [BCAA] were increased by 50% in the AA
group vs. C group (PG0.005). Glucose decreased in the AA group only
(22.6T1.5 baseline vs. 18.0T1.2 mg/dl final day, PG0.0005). Fetal arterial
plasma insulin, pH, pCO2, hematocrit, hemoglobin-O2 saturation, and blood
O2 content did not change. Fetal arterial blood pO2 decreased in AA from
baseline (18.9T0.7 mmHg) on days 5 (16.5T1.4 mmHg, PG0.05) and
8 (16.3T1.3 mmHg, PG0.05) then returned to baseline. Fetal arterial plasma
lactate concentrations increased in the AA group initially (2.05T0.13 baseline
vs 4.41T1.3 mmol/L on day 7, PG.005) then returned to baseline. The fetal
glucose/O2 quotient decreased in AA (PG0.05) but not in C.
Fetal glucose uptake was lower in AA vs. C (2.52T0.36 vs 3.86T
0.11mg/kg/min, PG0.05). Umbilical blood flow, lactate uptake, and O2
consumption rates did not change in AA fetuses.
Conclusions: A prolonged infusion of amino acids directly into fetal sheep
did not increase O2 consumption and preserved acid-base balance. We
speculate that decreased umbilical glucose uptake and glucose/O2 quotient in
the AA group is due to increased fetal amino acid oxidation.
294
INTRAUTERINE GROWTH RESTRICTION (IUGR)
INCREASES RETINAL EXPRESSION OF GENES INVOLVED
IN THE PATHOGENESIS OF RETINOPATHY OF
PREMATURITY (ROP) IN MALE RAT PUPS
Hale MA1, Jiang Y2, Yu X1, Callaway C1, Wang H2, Smith GT2, McKnight R1,
Lane RH1, Hartnett M2 1University of Utah, Salt Lake City, UT and 2University
of Utah, Salt Lake City, UT.
Purpose of Study: IUGR increases the risk of ROP in human preterm
infants. IUGR decreases human infant IGF-1 (insulin growth factor 1) serum
levels, which is a predictor of ROP. IUGR also affects IGF-1 serum levels in
rat pups. Despite the links between IUGR, ROP, and IGF-1, little information
exists on whether IUGR affects retinal IGF-1 expression. Retinal IGF-1 expression is relevant because 1) IGF-1 is expressed throughout the retina; 2)
IGF-1 mediates hypoxia inducible factor 1> (HIF-1>) expression; and 3)
IGF-1 induces erythropoietin (Epo) expression though HIF dependent and
independent mechanisms. Vitreous Epo expression is up regulated in patients
with vascular retinopathy. We hypothesized that IUGR increases IGF-1, HIF1>, and Epo mRNA levels, as well as levels of their respective receptors, in
newborn rat retinas.
Methods Used: To test this hypothesis, we induced IUGR in the rat pups
through uteroplacental insufficiency. Pups were killed at 7 days of life (75%
of retinal vascularization) (n = 6 each gender, control vs. IUGR). Retinal
cDNA was synthesized, and real time RT-PCR was performed. Target genes
include mRNA variants of IGF-1, IGF-1 receptor, HIF-1>, Epo, and Epo
receptor.
Summary of Results: In newborn male rat pups, IUGR significantly increased retinal mRNA levels of 1) IGF-1 mRNA variants from both the
promoter 1 and promoter 2 mRNA variants (P1: 172T24% of male controls;
P2: 245T47% of male controls) (p G 0.03); and 2) Epo mRNA levels
170
&
Volume 59, Number 1, January 2011
(144T17% of male controls) (p G 0.01). IUGR also increased HIF-1> and Epo
receptor mRNA levels, though significance was only approached (p = 0.06).
In newborn female rat pups, IUGR did not significantly affect gene
expression.
Conclusions: We conclude that IUGR increases retinal mRNA levels in
male newborn rats of genes associated with the pathogenesis of ROP. These
finding are intriguing considering previous reports associating retinal paracrine expression of IGF-1 and Epo with the development of vascular retinopathy in adults. We speculate that postnatal environmental insults induce a
neovascular retinopathy in IUGR male rat pups at a greater rate than controls
or female IUGR rat pups.
295
THE INHIBITORY POTENCY OF LOW DOSES OF ZINC
DEUTEROPORPHYRIN BIS GLYCOL ON HEME
OXYGENASE ACTIVITY IN 3-DAY-OLD MICE
Katayama Y, Shaw N, Yaffe ZA, He CX, Kalish FS, Schulz-Geske S, Zhao H,
Wong RJ, Stevenson DK Stanford University School of Medicine, Stanford, CA.
Purpose of Study: Neonatal hyperbilirubinemia is a common problem in
newborns. Structural heme analogs, metalloporphyrins (Mps), competitively
inhibit heme oxygenase (HO), the rate-limiting enzyme in bilirubin production, and are potential compounds for clinical use in the treatment of
neonatal hyperbilirubinemia. Although tin mesoporphyrin (SnMP) has been
studied in human neonates, its property as a photosensitizer and ability to
induce HO-1 may limit its clinical use. We have previously reported on the in
vivo efficacy of an alternative Mp, zinc deuteroporphyrin bis glycol (ZnBG)
in adult rodents. In this study, we investigated the efficacy of ZnBG towards
inhibiting HO activity and its effects on HO-1 expression in 3-day-old mice,
establishing a time and range of dosing for phototoxicity testing.
Methods Used: 3T0.5-day-old FVB mice were given vehicle (controls) or
ZnBG (0.12, 0.23, 0.47, and 0.94 Kmol/kg) IP. After 3h, mice were sacrificed
and the liver and brain were harvested and sonicated in phosphate buffer. HO
activity was quantified by gas chromatography, calculated as pmol CO produced/hr/mg fresh weight (FW) and expressed as % inhibition of HO activity
of control values. HO-1 expression was assessed by measurements of HO-1
protein using Western Blots.
Summary of Results: 3h after IP administration, ZnBG significantly
inhibited HO activity in the liver at all doses, ranging from 28% to 65%.
When % inhibition of HO activity was plotted against the dose of ZnBG, the
I50 of ZnBG was interpolated to be 0.325 Kmol/kg body weight. No significant inhibition of HO activity was observed in the brain. In addition, no
significant changes in HO-1 protein were found in either tissue for any dose.
Conclusions: We conclude that ZnBG is effective in inhibiting HO activity
rapidly and at relatively low doses in 3-day-old mice, and therefore, is an
attractive compound for potential use in the treatment of neonatal hyperbilirubinemia. Further study is required to determine the long-term effects of
ZnBG as well as its photoreactivity.
% Inhibition of HO Activity (meanTSD, n Q4), *pG0.01 vs control
296
BACTERIAL NITRATE REDUCTION IN SALIVA OF
NEWBORN INFANTS
Kanady JA1, Hopper AO1, Ninnis JR1, Aruni W2, Power GG3, Blood AB1,3
1
Loma Linda University, Loma Linda, CA; 2Loma Linda University, Loma
Linda, CA and 3Loma Linda University, Loma Linda, CA.
Purpose of Study: NEC is an acute, idiopathic disease characterized by
infection and necrosis of the bowel wall and occurs in approximately 5Y12%
of premature infants. Nitric oxide (NO) protects the normal adult GI tract by
increasing blood flow, thickening the mucous lining, and killing bacteria.
Nitrite is converted to NO under acidic conditions and therefore constitutes a
significant source of NO in the adult gastrointestinal tract. Swallowed nitrite
is derived by the action of anaerobic bacteria on the dorsal surface of the
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
tongue that convert ingested nitrates to nitrite. The activity of these bacteria in
the oral flora of newborn infants has not been studied. This study tests the
hypothesis that premature infants, known to be at increased risk for NEC,
have decreased bacterial reduction of nitrate to nitrite in the mouth.
Methods Used: Saliva was collected from preterm infants (n=8) and adults
(n=8) by swabbing the back of the tongue with a cotton-tipped applicator. The
volume of saliva collected was determined by weight gain of the swab, whose
tip was then immediately transferred to 3.0 mL of sterile, anaerobic broth at
37 -C. To assess nitrate-reducing activity, 333 mM nitrate was added to the
broth and the increases of nitrite concentration [NO2-] were measured over
30 min by triiodide chemiluminescence.
Summary of Results: Baseline salivary [NO2-] in preterm infants (0.91T
0.12 KM, SEM) was significantly lower than in adults (2.72T 0.35 KM;
pG0.001; ANOVA). The nitrate-reducing activity, calculated from the slope
of nitrite increase after normalizing to the volume of saliva collected, was
minimal in the saliva of preterm infants (2.6T2.7 nmoles/min/mg saliva), and
only a fraction of that in adult saliva (200T49 nmoles/min/mg; pG0.001).
Conclusions: The nitrite concentration in the saliva of preterm infants is
È33% of the salivary [NO2j] in adults. Nitrate reducing activity provided by
bacterial flora on the surface of the tongue of preterm infants is only È1.3%
of the activity found in adults. Thus preterm infants have far less nitrite to be
converted to NO than adults. This deficiency merits further study as a contributing cause of NEC.
297
PROLONGED NEONATAL INTENSIVE CARE STAY FOR
INFANTS OF DIABETIC MOTHERS
Phattraprayoon N1, Al-Ramadhani R2, Barton L1, Ramanathan R1 1Keck
School of Medicine, University of Southern California, Los Angeles, CA
and 2Keck School of Medicine, University of Southern California, Los
Angeles, CA.
Purpose of Study: There has been a decrease in morbidity and mortality
of infants of diabetic mothers (IDM) over the past decades, but there have
been few reports of reasons for IDM babies 937 weeks requiring longer stays
in the neonatal intensive care unit (NICU) than just to be monitored for hypoglycemia. We reviewed our IDM babies from 1997Y1999 and compared
them to a later cohort from 2007Y2009 to look at the length of stay, reasons
for NICU stay and to see if any improvement occurred over time.
Methods Used: IDM infants greater than A1 classification who came to our
NICU for hypoglycemia screening or other reasons during two time periods
(1997Y1999 and 2007Y2009) were reviewed for stays 9 24 hours to determine
the primary cause for continuing NICU care. We excluded babies G37 weeks
or with congenital anomalies because they would likely have other reasons
for NICU stay.
Summary of Results: Six hundred and thirty five IDM infants (348 born in
1997Y1999 and 287 in 2007Y2009) were reviewed for this study.
From the 1997Y1999 cohort, mean birth weight was 3595 T 586 g, range
2205Y5580 g and gestational age 38 T 1 weeks, range 37Y42 weeks, and
length of stay 1Y59 days.
From the 2007Y2009 cohort, mean birth weight was 3391 T 476 g, range
2220Y5040 g and gestational age 38 T 1 weeks, range 37Y42 weeks, and
length of stay 1Y64 days.
Incidence of hypoglycemia, large for gestational age (LGA), birth trauma
and length of stay were decreased in the later cohort; however, incidence of
respiratory distress and feeding intolerance did not improve.
Conclusions: IDM infants still remained at significantly increased risk of
requiring NICU care in our study. Major causes of length of stay have not
changed from earlier years. Respiratory distress and feeding intolerance are
still the main causes of prolonged NICU stay.
Results
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
298
NEONATAL MORTALITY RATE AND CAUSES OF DEATH IN
NINE VILLAGES IN THE LORETO PROVINCE OF PERU
Warren JB, Anderson JM Oregon Health and Science University,
Portland, OR.
Purpose of Study: Worldwide, 40% of deaths in children under-five years
of age occur in the neonatal period. The United Nations Millennium Development Goal 4 (MDG 4) calls for the reduction of the under-five mortality
rate by two thirds. Improvement in neonatal care worldwide is essential to
meeting MDG 4. In Peru, the 2008 neonatal (day 0Y28) mortality rate was 13
per 1000 live births with wide regional differences. The Loreto Province in
northeastern Peru is largely jungle and, in 2004, had an estimated early
neonatal (day 0Y7) mortality rate of 13 per 1000 live births. While national
mortality estimates are useful for global comparisons, knowing regional
neonatal mortality rates and causes of death is imperative to focus funding
and education for those areas most in need. Our objective was to determine
the neonatal mortality rate and causes of neonatal death for nine villages in
the Loreto Province.
Methods Used: We interviewed 131 women who were at least 15 years of
age and had been pregnant at least once in the last five years. Outcomes data
for each pregnancy were collected. In those identified as neonatal death, the
World Health Organization International Standard Verbal Autopsy for death
of a child under-4 weeks of age was used to determine cause of death.
Summary of Results: Outcomes data on 266 pregnancies were obtained.
Seven neonatal deaths were identified from 230 live births. Neonatal mortality rate was 30 per 1000 live births. The causes of neonatal death are listed
in the table. Infection was related to neonatal mortality in 57% of cases (4/7).
Conclusions: The neonatal mortality rate for nine villages in the Loreto
Province was 30 per 1000 live births, which is two times greater than the most
current estimate for all of Peru. The most common cause of or contributor to
death is infection. Knowledge of regional mortality rates and causes of death
is necessary to target interventions to reduce neonatal mortality.
299
A META-ANALYSIS OF DOPAMINE USE IN HYPOTENSIVE
PRETERM INFANTS: BLOOD PRESSURE AND CEREBRAL
HEMODYNAMICS
Sassano-Higgins S, Friedlich P, Seri I Childrens Hospital Los Angeles, Los
Angeles, CA.
Purpose of Study: Dopamine administration results in variable effects on
blood pressure in hypotensive preterm infants. The clinical benefits of dopamine administration in increasing cerebral blood flow (CBF) and reducing
adverse neurological outcomes in hypotensive preterm infants are unclear.
The current study was conducted to examine the efficacy of dopamine for
treatment of hypotension and to investigate the changes in cerebral hemodynamics and central nervous system injury in hypotensive preterm infants
following dopamine administration.
Methods Used: A meta-analytic design was used. All studies published in
peer-reviewed journals with sufficient information for statistical analysis were
included. Random and fixed effects models were used to calculate effect size
estimates and significance levels.
Summary of Results: Random effects meta-analysis found that dopamine increases mean arterial blood pressure (12 studies; N=163; r=0.88,
95%CI=0.76Y0.94) and systolic blood pressure (8 studies; N=142; r=0.81,
95%CI=0.42Y0.94). For increase in blood pressure, dopamine administration
was associated with a significantly greater overall efficacy than dobutamine
(7 studies; N=251; r=0.26; 95%CI=0.20Y0.32), colloid (2 studies; N=67;
r=0.60; 95%CI=0.41Y0.74), and hydrocortisone (1 study; N=28; r=0.40;
95%CI=0.034Y0.67). CBF increased following dopamine administration
171
Western Regional Meeting Abstracts
Journal of Investigative Medicine
(5 studies; N=75; r=0.36; 95%CI=j0.059Y0.67), and the increase in CBF was
greater in hypotensive than normotensive preterm infants (8 studies; N=153;
r=0.16; 95%CI=j0.0080Y0.32). There were no statistically significant differences in adverse neurological outcome between dopamine and dobutamine
(3 studies; N=118; r=j0.13; 95%CI=Y0.31Y0.059), epinephrine (2 studies;
N=46; r=0.06; 95%CI=j0.23Y0.34), colloid (2 studies; N=80; r=0.0070;
95%CI=j0.22Y0.23), or hydrocortisone (1 study; N=40; r=j0.10; 95%
CI=j0.40Y0.22).
Conclusions: Dopamine administration increases mean and systolic blood
pressure in hypotensive preterm infants, and is more effective than dobutamine, colloid, or hydrocortisone alone. Dopamine administration is associated with increased CBF, with greater increases in CBF in hypotensive than
in normotensive preterm infants. Dopamine is not associated with a greater
incidence of adverse effects than other therapies used to treat hypotension.
300
PRENATAL METHAMPHETAMINE USE AND NEONATAL
AND INFANT NEUROBEHAVIORAL OUTCOME: RESULTS
FROM THE INFANT DEVELOPMENT, ENVIRONMENT,
AND LIFESTYLE (IDEAL) STUDY
Kiblawi ZN1,2, Smith LM1,2, LaGasse LL2, Derauf C2, Newman E2, Shah R2,
Arria A2, Huestis M2, Haning W2, Strauss A2, DellaGrotta S2, Dansereau
LM2, Lester BM2 1Harbor-UCLA Medical Center, Torrance, CA and 2IDEAL
Community Research Network, Providence, RI.
Purpose of Study: Methamphetamine (MA) use among pregnant women is
an increasing problem in the United States. How MA use during pregnancy
affects neonatal and infant neurobehavior is unknown. The purpose of this
study is to examine the neurobehavioral effects of prenatal MA exposure in
the multicenter, longitudinal IDEAL study.
Methods Used: IDEAL screened 34,833 subjects at 4 clinical centers.
17,961 (67%) were eligible and consented, among which 412 were enrolled.
Exposed subjects were identified by self-report and/or GC/MS confirmation
of amphetamine and metabolites in infant meconium. Comparison subjects
were matched (race, birth weight, maternal education, insurance), denied
amphetamine use and had a negative meconium screen. Both groups included
prenatal alcohol, tobacco and marijuana use, but excluded use of opiates,
LSD, or PCP. The NICU Network Neurobehavioral Scale (NNNS) was administered within the first 4 days of life and again at 1 month old to 380
enrollees (185 exposed and 195 comparison). ANOVA tested exposure effects
on NNNS summary scores at birth and one month. GLM repeated measures
analysis assessed the effect of MA exposure over time on the NNNS summary
scores with and without covariates.
Summary of Results: Prenatal MA exposure was associated with decreased arousal (P=.001) and excitability (P=.043) scores relative to the
comparison newborns. By one month of age, both groups showed higher
quality of movement (P=.037), less lethargy (P=.001) and fewer asymmetric
reflexes (P=.016). Methamphetamine specific changes in NNNS scores over
the first month of life were noted with increased arousal (P=.031) and decreased total stress scores (P=.03).
Conclusions: Prenatal MA exposure in the neonatal period was associated
with neurobehavioral patterns of decreased arousal and excitability. Improvements in arousal and total stress were observed in MA exposed newborns by
one month of age.
301
EARLY PHYSICAL SIGN OF CONGENITAL RICKETS:
HARRISON_S THORACIC GROOVE
Chan GM University of Utah, Salt Lake City, UT.
Case Report: We would like to present two cases of congenital rickets
where the presence of Harrison_s groove was noted. Case 1 was a 34 week
gestation small for gestation female with a birth weight of 1100 g. This infant
was a product of a 33 year old gravida 7 para 4024. Mother noted decreased
fetal movement during the last 3 weeks of the pregnancy. Case 2 was a
30 week gestation appropriately sized female with a birth weight of 920 g.
This infant was a product of a 24 year old primigravida mother. Physical
exams during the first week of life was unremarkable except for the presence
of a linear groove or sulcus across the anterior lower part of the thorax in both
infants. Radiographic studies of the chest were reportedly normal for lungs
and bones. Laboratory findings included serum calcium levels of 10.9
172
&
Volume 59, Number 1, January 2011
and 10.6 mg% (normal 6Y10 mg%), phosphate of 2.3 and 4.0 mg% (normal 4.2Y9 mg%), 25-hydroxyvitamin D of 19 and 23 ng/mL (normal 30Y
60 ng/mL), 1,25-dihydroxyvitamin D of 107 pg/mL and 89 pg/mL (normal
25Y45 pg/mL), and urine calcium/creatinine ratio of 1.7 and 1.4 (normal
G0.7). We believe that the physical sign of the Harrison_s groove in these
infants is indicative of congenital rickets from maternal calcium and/or vitamin
D deficiency.
Nephrology and Hypertension
Concurrent Session
1:30 PM
Friday, January 28, 2011
302
IL-33-MEDIATED CISPLATIN-INDUCED AKI IS DEPENDENT
ON CD4 T CELL-MEDIATED PRODUCTION OF CXCL1
Edelstein C1, Akcay A1, Nguyen Q1, Turkmen K1, Lee D1, Faubel S1, He Z1,
Pacic A2, Galesic Ljubanovic D2, Jani A1 1University of Colorado, Aurora,
CO and 2University of Zagreb, Zagreb, Croatia.
Purpose of Study: The major complication of cisplatin administration is
acute kidney injury (AKI). IL-33 is a novel pro-inflammatory cytokine predominantly present in endothelium. We have previously demonstrated that
cisplatin causes endothelial cell apoptosis and necrosis. Thus the effect of
cisplatin-induced AKI on IL-33 was determined in mice.
Methods Used: Mice were injected with Cis 25 mg/kg. There is tubule
apoptosis (day 1), necrosis (day 2) and elevated BUN and serum creatinine
(SCr) (day 3).
Summary of Results: On immunofluorescence (IF) of kidney, IL-33 was
predominantly present in the endothelium. On immunoblot, Full-length
(active) IL-33 (34 kDa)increased in the kidney in cisplatin-induced AKI. On
ELISA, IL-33 increased in the kidney in cisplatin-induced AKI compared to
vehicle treated mice. To demonstrate the injurious role of IL-33, it was determined whether soluble ST2 (sST2), a fusion protein that neutralizes IL-33
activity by acting as a decoy receptor, protected against AKI or whether
injection of recombinant IL-33 (rIL-33) worsened AKI. Infiltration of CD4 T
cells into the kidney, serum creatinine (sCr), acute tubular necrosis (ATN) and
apoptosis decreased in the kidney in mice with cisplatin-induced AKI treated
with sST2. CD4 T cells, sCr, ATN and apoptosis were increased in wild type
mice, but not in CD4 j/j mice, injected with cisplatin plus rIL-33. Thus, the
injurious effect of IL-33 is mediated by CD4 T cells. CXCL1, a proinflammatory chemokine, is produced by CD4 T cells. CXCL1 was increased
in kidney in wild type, but not in CD4 T cell j/j mice treated with cisplatin
plus rIL-33. CXCR2 j/j mice, that are deficient in the receptor for CXCL1,
had lower sCr, ATN and apoptosis scores compared to wild type mice
with cisplatin-induced AKI, demonstrating the injurious role of CXCL1 in
cisplatin-induced AKI.
Conclusions: IL-33-mediated cisplatin-induced AKI is dependent on CD4
T cell-mediated production of CXCL1. Inhibition of IL-33 or CXCL1 has
therapeutic potential in cisplatin-induced AKI.
303
VITAMIN D INHIBITS CELL PROLIFERATION WITHOUT
INDUCING APOPTOSIS OF HEART-DERIVED CELLS,
ENHANCING CARDIAC DIFFERENTIATION
BY MODULATING THE EXPRESSION OF KEY Wnt
SIGNALING FAMILY MEMBERS
Garcia LA1, Ferrini M1,2, Gibbons G3, Shohet R4, Norris KC1,2, Artaza JN1,2
1
Charles R. Drew University of Medicine and Science, Los Angeles, CA; 2David
Geffen School of Medicine at UCLA, Los Angeles, CA; 3Morehouse School of
Medicine, Atlanta, GA and 4John A. Burns School of Medicine University of
Hawaii, Honolulu, HI.
Purpose of Study: Cardiovascular disease (CVD) remains the leading
cause of death in the US. Low levels of 25D (G15ng/ml) are associated with
high risk of myocardial infarction, even after controlling for factors known
to be associated with coronary artery disease. A growing body of evidence
suggests that the Vitamin D receptor and 1,25D (the active form of vitamin
D) plays an important role in several CVD related signaling pathways. The
Wnt signaling pathway is pivotal to gene expression and tissue development;
it has been shown to control stem cell renewal, lineage selection and more
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
importantly heart development. The purpose of this study is to demonstrate
that 1,25D induces cardiac differentiation by inhibiting cardiomyocytes cell
proliferation without promoting apoptosis through modulation of selected
members of the Wnt signaling pathway.
Methods Used: H9c2 cardiomyocytes incubated with or without 1,25D
were evaluated for cell proliferation, apoptosis, changes in cell area and the
expression of genes related to the cell cycle and Wnt family members by
RT2PCR, immunocytochemistry and westernblots.
Summary of Results: Addition of 1,25D to H9c2 cardiomyocytes: a)
promotes nuclear translocation of VDR; b) inhibits cell proliferation without promoting apoptosis; c) decreases the expression of genes related to the
regulation of the cell cycle; d) enhances cardiomyotubes formation; e) induces the expression of Casein kinase1->1 and; f) increases the expression
of Wnt11.
Conclusions: 1,25D promotes cardiac differentiation by negative modulation of the non-canonical Wnt signaling pathway through Casein kinase-1
and up-regulating the expression of Wnt11, which has been reported to induce cardiac differentiation. This is the first mechanistic demonstration that
supports the hypothesis that VD repletion might attenuate CVD not only by
down regulating excess cell proliferation but by promoting cardiac cellular
differentiation.
Western Regional Meeting Abstracts
were collected. The Kaplan-Meier product limit method was used to estimate
the survival rate. Univariate and multivariate survival analyses were performed using the Cox proportional hazard model.
Summary of Results: Graft failure occurred in 36 of the 98 patients
(36.7%) with a median time between Thymoglobulin treatment and graft
failure of 567 days (25Y75%: 152Y1265 days). In multivariate analysis,
factors associated with increased risk of graft failure included doses of
Thymoglobulin received (Q9) and number of biopsies performed on the
current allograft (Q1). Factors associated with decreased risk of graft failure
included creatinine level at time of Thymoglobulin treatment (G3.0 mg/dL)
and identification of vascular rejection versus non-vascular rejection on the
biopsy performed immediately prior to Thymoglobulin treatment.
Conclusions: Resolution of acute rejection is vital to allograft survival and
Thymoglobulin is a last-line treatment for steroid-resistant acute rejection.
Our results indicate that the doses of Thymoglobulin received, the creatinine level at time of Thymoglobulin treatment, the number of prior biopsies,
and the presence of non-vascular rejection are associated with increased
risk for graft failure. Individual chart-review permitted access to biopsy data
not available in database studies, and the analysis suggests that perhaps
Thymoglobulin treatment is less efficacious in acute rejection with chronic
changes. In patients with risk factors found to be significant for graft failure,
nephrologists may choose to lower or omit the use of Thymoglobulin as its
benefits may be outweighed by its recognized adverse effects in the long run.
304
COMPARISON OF METHODS FOR COUNTING PODOCYTES
Basgen J1, Nicholas S1,2 1Charles R. Drew University of Medicine and
Science, Los Angeles, CA and 2David Geffen School of Medicine at UCLA,
Los Angeles, CA.
Purpose of Study: The podocyte is a specialized epithelial cell that is
crucial in maintaining the structure and function of the glomerulus. Some
renal diseases report a decrease in podocyte number, but are inconsistent in
the podocyte number counted, possibly due to different counting methods. In
this study, we report results from three methods comparing podocyte number obtained from two widely used counting methods (Weibel-Gomez and
Fractionator/Disector) against a gold standard (Exhaustive Count).
Methods Used: Serial 1-Km epon sections (n=250) were cut from a single
mouse kidney. Images from all sections through nine glomeruli were used for
Exhaustive Count, which allowed all podocyte nuclei to be counted as they
were sequentially observed and is the gold standard. For the Weibel-Gomez
method the number of podocyte nuclei profiles were counted, and the volume
fraction of podocyte nuclei per glomerulus was determined using a counting
grid superimposed over the image. Shape and variance factors were determined and glomerular volume was measured followed by calculation of
podocyte number. For the Fractionator/Disector method, pairs of consecutive
sections were used to determine the number of podocytes. The number of
profiles from podocytes that were present in the Sampling Section but absent
in the Look-up Section was used to calculate podocyte number.
Summary of Results: From the Exhaustive Count method 84.3T7.7
(meanTSD) podocytes were counted compared with 89.3T6.8 podocytes
using the Weibel-Gomez method, which was significantly different, pG0.001.
From the Fractionator/Disector method, 81.9T13.3 were counted, with no
difference compared to the gold standard, p90.30.
Conclusions: These results suggest that the unbiased Fractionator/Disector
method is the recommended method for counting podocytes. Study was
funded by NIH grant 1U54RR026138.
305
FACTORS CONTRIBUTING TO THE EFFICACY OF
THYMOGLOBULIN AS A TREATMENT FOR ACUTE KIDNEY
GRAFT REJECTION
Liu Y1, Kuo H1, Jiang Y2, Bunnapradist S1 1UCLA Medical School,
Los Angeles, CA and 2UCLA, Los Angeles, CA.
Purpose of Study: To examine the outcomes of Thymoglobulin treatment
for acute rejection and to identify risk factors associated with graft failure
using the UCLA Kidney Transplant Database.
Methods Used: Between February 2000 and June 2010, there were 517
hospital admissions for possible kidney allograft rejection. Among these,
there were 114 unique patients treated with Thymoglobulin for acute rejection, and 98 were included in the analysis, with a median follow up time of
567 days (16 patients did not have sufficient follow-up data). An analytical
file was created by retrospective chart-review and baseline characteristics
* 2011 The American Federation for Medical Research
306
HOSPITALIZATIONS IN ELDERLY
HEMODIALYSIS PATIENTS
Martinez MA1,2, Sun Y1,2, Servilla K1,2, Murata GH1,2, Tzamaloukas AH1,2
1
University of New Mexico Health Sciences Center, Albuquerque, NM and
2
Raymond G. Murphy VA Medical Center, Albuquerque, NM.
Purpose of Study: There is an increasing amount of elderly patients developing end-stage renal disease. The benefit of chronic hemodialysis (HD)
for elderly patients has been questioned. Prior studies have not been consistent on whether age is a predictor of hospitalization in chronic HD patients.
The purpose of the study was to determine whether hospital admissions and
hospital days differed between elderly and younger patients on chronic HD.
Methods Used: Hospitalization rate (HR), length of hospitalization (LH)
and co-morbidities were compared between patients 970 years (Group A)
and G70 years (Group B). HR is compared in number per patient per year
(n/[pt-yr]) and LH in days per patient per year (d/[pt-yr]), both presented
as Mean (95% Confidence Interval) between two groups of incident HD
patients.
Summary of Results: Group A had initiation of HD at 77T5 years (n = 173).
Group B had initiation of HD at 57T8 years (n = 213). Diabetes mellitus
(DM) was present in 50% of the Group A patients and 65% of the Group B
patients. Differences in hospitalizations from all causes were marginal: rate,
Group A 2.46 (2.11, 2.81), Group B 1.97 (1.74, 2.20) n/[pt-yr], p = 0.061;
length, Group A 34.2 (26.3, 42.1), group B 23.2 (18.1, 28.2) d//[pt-yr],
p = 0.053. Hospitalizations for individual categories (e.g. cardiac, HD vascular access, infections, etc) did not differ between the two groups. Among
patients with DM, group A subjects had a higher admission rate and greater
length of hospitalizations from all causes and from infections than Group B
subjects. No difference was found for any category of hospitalization between Group A and Group B patients without DM. For hospitalizations from
all causes, multiple linear regression identified co-morbidity, quantified by
the Charlson index (p G 0.001), poor compliance with the HD schedule
(p = 0.069) and only marginally advanced age at HD initiation (p = 0.092)
as predictors of a high rate, and high Charlson index (p G 0.001), older age
at HD (p = 0.035), and marginally poor compliance (p = 0.069) as predictors
of great length.
Conclusions: Co-morbidities, which includes DM, are the most potent
determinants of frequency and length of hospitalizations in HD patients,
while advanced age has only marginal effects.
307
THE ROLE OF NETRIN-1 RECEPTORS IN MEDIATING
ATTENUATION OF ACUTE KIDNEY INJURY
Luebbert TM1, Dalton J1, Grenz A2, Eltzschig HK2 1University of Colorado
School of Medicine, Aurora, CO and 2University of Colorado School of
Medicine, Aurora, CO.
173
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Purpose of Study: Ischemia is the most common cause of acute kidney
injury (AKI). For example, patients undergoing surgical procedures requiring cross-clamping of the aorta or renal vessels experience AKI in up
to 30% of cases. Similarly, AKI after cardiac surgery occurs in approximately 10% of patients and is associated with dramatic increases in morbidity and mortality. Our laboratory has previously shown that netrin-1
attenuates kidney injury and inflammation following renal ischemia. The
current study will investigate which of the known netrin-1 receptors
(UNC5A, UNC5B, UNC5C, UNC5D, neogenin, DCC, A2BAR) mediates
renal tissue protection. These findings could provide new therapeutic options for patients with AKI.
Methods Used: Mice were anesthetized using intraperitoneal pentobarbital. Mice then underwent renal ischemia (30 or 60 minutes) using a
previously-described hanging-weight system to selectively occlude the left
renal artery. The kidney was allowed to reperfuse (2 or 6 hours), then left
nephrectomy was perfomed and the kidney was flash-frozen. Each experiment was completed on 3 wild-type and 3 mice with partial depletion of
netrin-1 expression (Ntn1+/j). Homozygote mice gene-targeted for netrin1 are not viable and die shortly after birth. Fold-changes in netrin receptor
expression were determined via RT-PCR. Confirmatory studies will be
completed using Western blot for receptors that show significant foldchanges in mRNA expression.
Summary of Results: Preliminary data show an up-regulation of
UNC5B and A2BAR in wild type mice and heterozygote mice following renal ischemia and reperfusion compared to controls without ischemia. We have not shown any regulation of the other receptors so far.
However, the number of experiments must be increased to confirm this
initial data set.
Conclusions: Based on mRNA analysis, the kidneys seem to up-regulate
the UNC5B and A2BAR after renal ischemia. Further studies must be performed to confirm these preliminary findings.
308
RISE IN SERUM SODIUM CONCENTRATION AFTER
SALINE INFUSION IN HYPONATREMIA
Rosen BH1, Malhotra D2, Murata GH1, Raj DS3, Shapiro JI2, Tzamaloukas A1
1
University of New Mexico, Albuquerque, NM; 2University of Toledo, Toledo,
OH and 3George Washington University, Washington DC, DC.
Case Report: We investigated factors that may lead to quantitative discrepancies between predicted and measured serum sodium [Na] in a patient
who developed osmotic myelinolysis.
A 55-year-old man with alcoholism was admitted with diarrhea and repeated falls. Blood pressure was 93/49 mm Hg and pulse 90 bpm. Neurologic, cardiac and pulmonary examinations were normal. Edema was not
detected. Initial [Na] was 111 mmol/L and serum osmolality 231 mOsm/kg.
Six hours later, after infusion of 1.75 L normal saline (NS), [Na] was
120 mmol/L, serum osmolality 254 mOsm/kg, urine osmolality 74 mOsm/kg
and urine sodium 10 mmol/L. Infusion rate was decreased. After another
12 hours and a total of 2.5 L NS, [Na] was 129 mmol/L. Osmotic myelinolysis manifested two days later.
To compare values observed and those predicted by the Adrogue-Madias
formula for an infusion of 1.75 L NS, we classified the potential pitfalls into
three categories: (A) Estimates entered in the formula. Volume and sodium
concentrations in infusate and serum are measured, but body water is estimated. We varied body water by 10 L (36 and 26 L) with resulting predicted
serum [Na] of 113.0 and 113.7 mmol/L. (B) Omission of other factors affecting [Na]. Considering changes in the osmotic coefficients of sodium and
potassium and the effects of the Gibbs-Donnan equilibrium, the predicted
[Na] for body water of 36 and 26 L is 133.2 and 114 mmol/L. (C) Assuming
a closed system. We calculated the effects of external losses of water, sodium and potassium through the lungs, skin, gastrointestinal system and the
kidneys. An estimated loss of 0.3 L of water through the first three routes
would lead to a [Na] of 114.7 and 116 mmol/L for body water values of
36 and 26 L. The calculated urine volume with sodium and potassium concentrations of 10 mmol/L each, needed to raise the [Na] to 120 mmol/L was
2.0 and 1.1 L, respectively.
External losses of water and solute are the main causes of discrepancies
between predicted and final [Na] values in patients treated for hyponatremia.
As these losses are unpredictable, monitoring of clinical status and laboratory
parameters during saline infusion is imperative.
174
&
Volume 59, Number 1, January 2011
Western Student Medical Research Forum
Student Session IV - Developmental Biology,
Genetics, and Hematology Oncology
1:30 PM
Friday, January 28, 2011
309
MAINTENANCE OF PHENOTYPE OVER SERIAL PASSAGES
IN FETAL CELL CULTURES
Dye AM, McConaghy S, Ohls R UNM, Albuquerque, NM.
Purpose of Study: Obtaining fetal samples at specific gestational ages
and in timely fashion for investigation can be problematic. The possibility of
maintaining the phenotype of a variety of primary human fetal cell cultures
through passing and freeze/thawing has not been evaluated. In addition,
smaller samples at younger gestational ages limit the total cell number to be
studied, while growing cells under stable conditions allows expansion of cell
numbers. We sought to determine if cell phenotype could be maintained in
retinal and brain cell cultures following freezing and reconstitution.
Methods Used: Human fetal retinal and brain samples were collected between 15Y23 weeks of gestation. Single cell suspensions were created and
suspended in Delbecco_s Modified Essential Media, with 10% fetal bovine
serum and 1% antimicrobials added. Cells were grown to confluence and
passed by incubating in 0.05% trypsin, washing in DMEM, and resuspending at a concentration of 5 105 cells per flask. Cells were passed twice
before freezing. Cells were grown on coverslips with each passage, and fixed
with 3.7% formalydehyde for 5 minutes, then stored in phosphate buffered
saline for later immunostaining. Cells were removed from tissue flasks with
trypsin, washed and pelleted, and resuspended in 1 mL of 7.5% dimethyl
sulfoxide (DMSO) in DMEM. The concentrated cell suspension was slowly
frozen in an ethanol freezing chamber. Cells remained frozen at j80- Centigrade for 24 hours, then transferred to liquid nitrogen (j196- Centigrade).
Coverslips were stained with antibodies directed against GFAP (astrocytes),
Nestin (neuronal stem cells), or MAP-2 (neurons).
Summary of Results: Both neuronal and retinal cell lines were viable
after 2 passages and a freeze-thaw step. Cells remained frozen for a minimum
of 3 weeks, and a maximum of 5 months. In all cases, cell lines were successfully reconstituted to the next stage without significant change in phenotype. In retinal cell primary cultures, astrocytes comprised 92Y94%,
neuronal stem cells comprised 3Y5%, and neurons comprised 1Y2% of total
cells. In brain cell primary cultures, cells consisted of 80Y85% astrocytes,
5Y8% stem cells, and 3Y4% neurons.
Conclusions: Both retina and brain cultures were successfully reconstituted.
Further studies are required to determine if the phenotype of the cells change
with further passages.
310
DISPARATE CONSERVATION OF GENE EXPRESSION IN
LMX1B-DIRECTED LIMB DORSALIZATION
Estes MK, Feenstra JM, Pira CU, Oberg KC Loma Linda University, Loma
Linda, CA.
Purpose of Study: The role of LMX1B in limb dorsalization is well
established; however, the process by which it regulates the cascade of events
required for dorsalization remains unclear. Identification of downstream
targets of LMX1B is imperative for understanding limb development and
managing dysmorphogenic diseases such as Human Nail Patella Syndrome.
Microarray analysis comparing wild-type and Lmx1b knockout embryonic
mouse limbs (11.5, 12.5, and 13.5 days post coitum) identified several potential downstream gene targets of Lmx1b. Utilizing whole mount in situ
hybridization (WMISH) analysis, four particular genes of interest (Keratocan,
Lumican, Decorin, and Emx2) have shown dorsally restricted expression in
the mouse model. We hypothesized that Lmx1b-restricted dorsal expression
of these target genes would be conserved throughout tetrapod limb development. Demonstration of conserved developmental expression would allow
clinical questions to be evaluated in animal models. Characterization of normal expression of these potential target genes in animal models is necessary
to characterize their regulation by Lmx1b.
Methods Used: Using WMISH, normal expression patterns for LMX1B,
KERATOCAN, LUMICAN, DECORIN, and EMX2 were documented in the
chick at Hamburger-Hamilton stages 21, 23, 25, and 27.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Uniquely individual expression patterns were observed for all four target genes; however, none of these molecules displayed
dorsally restricted expression patterns. Although initiation of expression
occurred at a stage later than LMX1B, their expression patterns included
specifically localized dorsal and ventral regions. Interestingly, each of the
genes studied demonstrate an overlap of expression in the presumptive
scapular region indicating that LMX1B may utilize these genes to pattern
the scapula.
Conclusions: The lack of correlation between chick and mouse expression
patterns suggests that the LMX1B-directed gene cascade of limb dorsalization is not tightly conserved across species.
311
INCREASED LEF1 EXPRESSION IMPLICATES THE
WNT/A-CATENIN PATHWAY IN THE MEDIATION OF
FGF-INDUCED UP-REGULATION OF SHH
Mattison GL, Steinman ED, Pira CU, Oberg KC Loma Linda University,
Loma Linda, CA.
Purpose of Study: Shh and Fgf play critical roles in limb development and
regeneration. Fgf is secreted from the apical ectodermal ridge (AER) of the
developing limb and promotes outgrowth. In addition, AER-related Fgf
maintains Shh expression from posterior mesoderm adjacent to the AER via
a reciprocal Shh/Fgf loop. In Fgf-induced chick limb regeneration, Shh is
up-regulated; however, the mechanism of Fgf-induced Shh up-regulation is
unknown. Gene array data, derived at 24 hrs during Fgf-induced Shh upregulation, suggested that the Wnt/A-Catenin pathway might play a role
in this mechanism. Thus, we hypothesized that Lef1, a downstream transcription factor of the Wnt/A-Catenin pathway, might be up-regulated by
Fgf during Shh induction.
Methods Used: To determine Lef1 involvement in Shh induction and the
potential mechanistic sequence of events, we generated a Lef1 expression
time course after exogenous Fgf application. Fgf-soaked heparin-acrylic
beads were implanted into the posterior limb bud of Hamburger-Hamilton
stage 23 chick embryos, harvested 3Y24 hrs later, and then analyzed using
whole mount in situ hybridization (WMISH) for Lef1 expression.
Summary of Results: We confirmed the gene array data at 24 hrs with
robust Lef1 up-regulation in response to Fgf. At 3 and 6 hours, WMISH
showed no significant Lef1 up-regulation. By 12 hours, there was detectable
expression that progressively increased at 18 and 24 hours.
Conclusions: In previous studies, we found other Wnt/A-Catenin pathway genes, for Wnt5a and its receptor Fzd4, to be up-regulated at 12 and
18 hours (respectively) after Fgf bead implantation. In this study, we found
that Lef1, the gene for a downstream transcription factor of Fzd4, was upregulated within the same period as Wnt5a and Fzd4. Collectively, this
data further implicates the Wnt/A-Catenin pathway during Fgf induced upregulation of Shh.
312
DEFINING THE MECHANISM OF LIMB REGENERATION:
A POTENTIAL NOVEL ROLE FOR BAMBI IN MEDIATING
FGF-INDUCED SHH UP-REGULATION
Steinman ED, Mattison GL, Pira CU, Oberg KC Loma Linda University,
Loma Linda, CA.
Purpose of Study: Shh is critical to limb patterning during development
and regeneration. The apical ectodermal ridge secretes Fgfs that induce
limb outgrowth and maintain Shh expression in the zone of polarizing activity through a reciprocal Fgf-Shh loop. However, the mechanism by which
Fgf regulates Shh expression is unknown. To identify intermediate molecules involved in this mechanism, we performed gene arrays 24 hr after FGF2
application to the posterior region of the chick limb bud. The gene array data
demonstrated elevated levels of BMP pathway associated genes (BMP7 and
BAMBI). We hypothesized that the BMP pathway might participate in the
Fgf-induced up-regulation of Shh.
Methods Used: We analyzed the expression time course of BMP7 and
BAMBI during FGF-induced SHH up-regulation. FGF2-soaked beads were
implanted into the posterior aspect of stage 23 chick wing buds. The chicks
were then harvested after 3Y24 hr of incubation and an in situ hybridization
for BMP7 and BAMBI was performed.
Summary of Results: BMP7 expression was not elevated after 24 hr of
FGF2 treatment (in contrast to gene array data), while FGF2-induced BAMBI
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
up-regulation was validated. Furthermore, FGF2 elevated BAMBI expression
within 6 hr after application.
Conclusions: These results were unexpected. We anticipated that BMP7
would be up-regulated, followed by its inhibitor, BAMBI. The up-regulation
of BAMBI without up-regulation of BMP7 suggested an alternative role for
BAMBI. Recent studies have demonstrated that, in addition to its role as a
BMP inhibitor, BAMBI can act as a positive modulator of the Wnt/A-catenin
pathway, a pathway known to participate in Shh regulation. Our data suggests
that BAMBI plays a novel role in Fgf-induced Shh up-regulation by modulating the Wnt/A-catenin pathway.
313
ASSESSING SUSCEPTIBILITY TO AGE-RELATED MACULAR
DEGENERATION WITH GENETIC MARKERS AND
ENVIRONMENTAL FACTORS
Grob S1,2,3, Chen Y3, Zeng J3, Zhou X3, Bernstein P2, Bedell M2,3,
Goldbaum M2, Ferreyra H2, Kozak I2, Freeman W2, Zhang K2,3 1University
of California at San Diego, School of Medicine, La Jolla, CA; 2Shiley Eye
Center, La Jolla, CA and 3University of California, San Diego, La Jolla, CA.
Purpose of Study: To evaluate the independent and joint effects of genetic
factors and environmental variables on advanced forms of age related macular degeneration (AMD) including geographic atrophy (GA) and choroidal
neovascularization (CNV) and to develop a predictive model with both genetic and environmental factors included.
Methods Used: Demographic information, including age of onset, smoking
status and BMI, was collected in 1844 participants. Genotypes were evaluated for eight variants in five genes related to AMD. Unconditional logistic
regression analyses were performed to generate a risk predictive model.
Summary of Results: All genetic variants showed strong association with
AMD. Multivariate odds ratios (ORs) were 3.52 (95% CI: 2.08Y5.94) for
CFH rs1061170 CC, 4.21 (95% CI: 2.30Y7.70) for CFH rs2274700 CC, 0.46
(95% CI:0.27Y0.80) for C2 rs9332739 CC/CG, 0.44 (95% CI: 0.30Y0.66) for
CFB rs641153 TT/CT, 10.99 (95% CI: 6.04Y19.97) for HTRA1/LOC387715
rs10490924 TT and 2.66 (95% CI: 1.43Y4.96) for C3 rs2230199 GG. Smoking
was independently associated with advanced AMD after controlling for age,
gender, body mass index (BMI) and all genetic variants.
Conclusions: CFH confers more risk to the bilaterality of GA whereas
LOC387715/HTRA1 contributes more to the bilaterality of CNV. Risk
models with combined genetic and environmental factors together has a
notable discrimination power; Further C3 confers more risk for GA than
CNV. Targeting high risk individuals for surveillance and clinical interventions may help reduce disease burden.
Clinical Relevance: Early detection and risk prediction of AMD could
help to improve the prognosis of AMD and reduce the outcome of blindness.
314
EXCLUSION OF PATHOGENIC CODING REGION
MUTATIONS IN POSITIONAL CANDIDATE GENES FOR
POSTERIOR AMORPHOUS CORNEAL DYSTROPHY
Kim MJ1,2, Lai IN2, Nguyen CK2, Rayner S2, Yellore V2, Aldave AJ2
1
David Geffen School of Medicine at UCLA, Los Angeles, CA and 2Jules
Stein Eye Institute, Los Angeles, CA.
Purpose of Study: Posterior amorphous corneal dystrophy (PACD) is a
rare autosomal dominant disorder that is associated with a constellation of
disorders of the anterior segment of the eye. Clinical findings include partial
or complete posterior lamellar corneal opacification, decreased corneal
thickness, scleralization of the peripheral cornea, corneal flattening with an
average corneal curvature less than 41 D, iridocorneal adhesions, correctopia,
and iris atrophy. Genome-wide linkage analysis has demonstrated linkage
to chromosome region 12q21.33. There are 22 genes in the linked interval,
4 of which were previously found not to have pathogenic coding region
mutations. The purpose of this project was to screen the coding regions of
the remaining 18 genes to identify the genetic basis of PACD.
Methods Used: PCR amplification of the exonic regions of each positional
candidate gene was performed using DNA from two affected individuals, two
unaffected individuals, and an unrelated control. Sanger sequencing reactions
were carried out, and variants were identified by comparison of the generated
and wild type sequences.
Summary of Results: Screening of 16 of the 18 positional candidate genes
(ATP2B1, BTG1, C12orf12, CENPC2, CLLU1, CLLU1OS, DUSP6, GALNT4,
175
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Loc100132126, Loc100287355, Loc100287398, Loc100287505, Loc338758,
MRPL2P1, MRPS6P4, and WDR51B) has been completed. Thirty eight sequence variants were identified in 10 genes, including 35 known variants and
3 novel variants. 10 silent mutations, 8 missense mutations, 1 nonsense mutation, 1 deletion, and 18 non-coding RNA changes were identified. None of the
variants segregated with the affected phenotype in the family.
Conclusions: The coding regions of 20 of the 22 positional candidate genes
for PACD have not revealed the pathogenic mutation in the family in which
linkage to chromosome region 12q21.33 was demonstrated. The pathogenic
mutation may be contained in the two remaining genes in the linkage interval
(Loc643153 and RPL21P106), which are currently being screened. Alternatively, the mutation may be located slightly outside the linkage interval,
may involve a copy number variant or may be in a non-coding region of one
of the genes already screened.
315
ENHANCED LEUKOCYTE CHEMOTAXIS INTO SERUM
FROM A MOUSE MODEL OF GAUCHER DISEASE
Jabre NA2, Pandey M1, Grabowski G1 1Cincinnati Children’s Hospital,
Cincinnati, OH and 2University of Cincinnati, Cincinnati, OH.
Purpose of Study: Gaucher Disease is a lysosomal storage disorder
resulting from mutations in the gene encoding glucocerebrosidase, an enzyme that cleaves glucose from Glucosylceramide (GC). Defective activity
leads to accumulation of GC within macrophages. Affected individuals exhibit hepatosplenomegaly, and CNS deterioration in certain subtypes. Pathologic changes may be attributable to stimulation of macrophages by excess
GC, leading to over-secretion of cytokines and subsequent activation of other
inflammatory leukocytes. To determine if the cytokine profile and leukocyte
activity are abnormal in Gaucher Disease, we analyzed the cytokine contents
of serum from a mouse model of the disorder (9V), and measured the chemotactic response of WT and 9V neutrophils and lymphocytes to both WT
and 9V serum.
Methods Used: Pooled serum was obtained from WT (n=14) and 9V
(n=17) mice and analyzed for the presence of leukocyte-specific cytokines
using a cytokine array. Neutrophils and Lymphocytes from WT and 9V
spleen and lung tissue were then isolated and analyzed by flow cytometry
to confirm identity. The serum and cells were placed on opposing sides of a
3-5Hm porous membrane in a Boyden Chemotaxis Chamber, and were
incubated at 37C for 30min to allow cell migration. Following incubation,
the membrane was removed and stained, and the number of migratory cells
was counted.
Summary of Results: Cytokine analysis of WT and 9V serum indicated
elevated levels of neutrophil (GM-CSF, IL-17), B-Cell (CXCL13, IL-10), and
T-Cell (IL-7, CXCL9, SDF-1, IL-27, IL-16, IL-12p70) specific cytokines/
chemokines in 9V vs. WT serum. Migration of 9V splenic neutrophils and
lymphocytes into 9V serum was characterized by a 4.2 and 1.6-fold increase
in cell number as compared to migration of WT splenic neutrophils and
lymphocytes into WT serum. Similarly, migration of 9V lung neutrophils and
lymphocytes into 9V serum exhibited a 12.6 and 5.7-fold increase, respectively, as compared to their WT counterparts.
Conclusions: Enhanced migration of leukocytes into 9V serum as compared to WT serum is likely due to the presence of an elevated cytokine
profile in the 9V model. Furthermore, 9V leukocytes may be Bactivated[
and more likely to respond to the presence of cytokines than their WT
counterparts.
316
FAMILIAL IDIOPATHIC SCOLIOSIS: INVESTIGATION
OF SFI1 GENE IN ADOLESCENT MALES WITH SEVERE
CURVATURES
Smith GA, Dunn JH, Swindle K, Miller NH University of Colorado School
of Medicine, Aurora, CO.
Purpose of Study: Idiopathic Scoliosis (IS) is a structural lateral curvature
of the spine Q10 degrees that occurs in 2Y3% of individuals of which 3Y9%
require active treatment. Genetic studies suggest that IS is a complex genetic
disorder, with multiple genes and genetic variations contributing to its expression. Genomic screening and subsequent fine mapping has resulted in
significant association between a subgroup of families characterized by a
male having a Q30 degree curvature and an area on chromosome 22 inclusive
176
&
Volume 59, Number 1, January 2011
of the SFI1, SF3A1 and LARGE genes. The goal of this study is to examine
further this association through sequencing analysis of the SFI1 gene within
this subpopulation.
Methods Used: DNA was extracted from blood samples of families with
two or more individuals affected by IS (202 families: 1198 individuals). A
subgroup of trios comprised of parents, regardless of curvature status, and
affected son(s) (18 families, 56 individuals) with curvatures Q30 degrees
were identified for sequencing. Curvature status of parents and son(s) was
confirmed by X-ray and noted for analysis. Primers were designed using
Primer Express\ software to amplify and sequence the 32 coding exons of
SFI1. Standard Sanger sequencing methods were used with read lengths of
~900 nucleotides.
Summary of Results: Results are contingent on ongoing sequencing
analysis. Anticipated results will show any significant associations between
IS and the SFI1 gene.
Conclusions: The current work builds on previous data indicating a significant association between the SFI1 gene and a phenotypic subset of families with IS. The use of clinical criteria to characterize the scoliosis
phenotype may aid in decreasing the heterogeneity of any one large study
population, and enhance the successful identification of specific genes responsible for this disorder. SFI1 is involved in spindle assembly and is
translated in several tissues including bone, nerve, and connective tissues
with notable protein interactions relating to bone and connective tissue development. The identification of a genetic locus is of major clinical and
therapeutic interest and may allow for a deeper understanding of spinal
growth and stability.
317
ASSESSING INTEREST AND ATTITUDES TOWARD
GENETIC PREDICTIVE TESTING FOR COLON CANCER:
METHODOLOGY AND INITIAL PREDICTORS
Tuong W1, Leventhal K2, Graves K2 1UC Davis, Sacramento, CA and
2
Georgetown University, Washington, DC.
Purpose of Study: With continued expansion of personalized medicine
through both commercial availability of predictive SNP testing and emerging
clinical translation of certain genomic applications, we sought to assess interest in, attitudes toward, and initial predictors of SNP testing in a primary
care population.
Methods Used: Based on prior data, we developed hypothetical scenarios
to assess interest in SNP testing. We assessed demographics, interest in
learning about genetic testing, genetics knowledge, and positive/negative
attitudes about SNP testing. We conducted cognitive testing of the survey
with 42 primary care patients.
Summary of Results: Participants (n=19 men, 23 women) reported a mean
age of 50.6 (SD=16.2 years). Most participants (74%) reported they would
Bprobably[ or Bdefinitely[ have a colon cancer SNP test. Greater interest in
SNP testing was related to more interest in learning about genetic testing
(1^2=10.14, p=0.001), and having both higher positive (t=j2.258, df=33,
p=0.031) and negative attitudes (t= j2.314, df=33, p=0.027) toward SNP
testing. Further, participants with higher genetics knowledge had more positive attitudes toward testing (t=j2.260, df=33, p=0.024). In this initial exploration, demographics, genetics knowledge, and personal and family
history of cancer were unrelated to interest in SNP testing.
Conclusions: Primary care patients indicated strong interest in SNP testing
and in learning more about genetic testing. Interestingly, both positive and
negative attitudes toward SNP testing were related to a greater desire to have
SNP testing. Future development of educational tools may help patients
clarify the pros and cons of SNP testing in order to promote more informed
decisions about personalized genomic medicine.
318
FACTOR VIII (FVIII:C) LEVELS WITH AGE IN A COHORT
OF INDIVIDUALS WITH MILD HEMOPHILIA A
Lee L1, Ludlow A2, Deb G2, Vickars L2, Wu J3, Jackson S2 1UBC,
Vancouver, BC, Canada; 2St. Paul’s Hospital, Vancouver, BC, Canada and
3
BC Children’s Hospital, Vancouver, BC, Canada.
Purpose of Study: Hemophilia A is a disorder resulting in absent or reduced FVIII:C levels. Baseline FVIII:C levels are important in determining
management in mild hemophilia (FVIII:C 0.06Y0.40IU/mL). In healthy individuals, FVIII:C increases with age. This trend has not been well characterized
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
in mild hemophilia. This is important to consider because overtreatment
with factor concentrate may paradoxically increase the thrombosis risk.
The study objectives are to describe the changes in baseline FVIII:C levels
with age in a cohort of subjects with mild hemophilia and to identify any
factors (blood type, co-infection status, FVIII mutation type) which may affect
FVIII:C levels.
Methods Used: Medical records for subjects with FVIII levels 0.060.40IU/mL registered with the BC Provincial Bleeding Disorder program
were reviewed. Subjects with a minimum of 2 FVIII:C levels at least 5 years
apart were included in the linear mixed effects model. Retrospective data
was extracted from medical records including age, sex, blood type, FVIII mutation, historical FVIII:C levels, DDAVP response, and co-infection (Hep.C,
B and HIV) status. Hypothesis testing was applied to estimate the effects of
co-variates on the variability and baseline FVIII:C.
Summary of Results: 137 subjects with mild hemophilia were reviewed
for eligibility. 83 subjects were excluded due to charts not available (n=27)
or inadequate FVIII:C data (n=56). Median age was 49 yrs (IQR 33, 59). All
subjects were male. Median duration of observation was 15.8 yrs (IQR 9.4,
23.9). Linear mixed effects modeling using 3 different models showed mean
trend of zero. There were fewer data points for subjects over the age of 50 as
compared to other age groups, limiting the ability to assess FVIII:C trends in
older subjects. There was no observable effect of the co-variates assessed on
baseline or variability of FVIII:C.
Conclusions: Our study showed no trend in changing baseline FVIII:C
levels with age in individuals with mild hemophilia. Co-variate effect on
baseline FVIII:C levels and variability was not observed. Further analysis
using similar methods with data from additional hemophilia centres, particularly in subjects 950 yrs, is necessary to confirm these observations.
319
SURVIVAL OUTCOMES OF ADULTS WITH ACUTE MYELOID
LEUKEMIA - A STUDY OF NATIVE AMERICAN, HISPANIC,
AND NON-HISPANIC WHITES TREATED IN
NEW MEXICO (1986Y2006)
Rivera N1, Armijo B1, Quintana D2, Abdul-Jaleel M1, Lee S3, Wiggins C2,
Libby E2 1UNM- School of Medicine, Albuquerque, NM; 2University of
New Mexico Cancer Center, Albuquerque, NM and 3University of New
Mexico, Albuquerque, NM.
Purpose of Study: New Mexico is composed of three main ethnic populations Caucasians, Native Americans, and Hispanics. As the largest
tertiary care center in the state, we offer treatment to significant numbers
of all three groups. Also, many residents of the state live in rural areas.
The goal of this study is to identify potential differences in outcome between ethnic groups from urban or rural areas in patients with newly diagnosed acute myelogenous leukemia (AML) treated at this center from
1986Y2006.
Methods Used: A retrospective chart review was performed on adult
patients with a new diagnosis of AML treated from 1986 through 2006. 144
charts were available for review. Data was collected for race/ethnicity, sex,
zip code, cytogenetics, treatments received, total number of chemotherapy
cycles, achievement of complete remission, and overall survival from time of
diagnosis and from achievement of remission after induction chemotherapy.
Date of death was determined from New Mexico tumor registry data. Statistical analysis was performed to examine the effect of the variables mentioned above on overall survival.
Summary of Results: The study included 144 patients (mean age 49.98,
range 18Y85). Patients who received standard treatment of 7+3 followed by
high dose cytarabine (1Y4 cycles) or similar regimen; there were no statistical
differences between survival rates when comparing the three ethnic groups,
Caucasians 78 (52%), Hispanics 43 (28%), and Native Americans 23 (15%)
(P=0.2496). In agreement with previous reports, statistically significant differences were seen for treatment versus palliative care, cytogenetics and age.
The study included 61 urban patients with a mean survival of 426 days and
83 rural patients with a mean survival of 278 days. Analysis revealed a
statistically significance in survival for urban versus rural patients who
achieved complete remission after induction chemotherapy (P=0.0443).
Conclusions: Survival was improved for urban patients who achieved a
complete remission after induction chemotherapy vs rural patients (P=
0.0443). Further study should be undertaken to identify the causes of this
disparity.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
320
NOVEL COMBINATIONS OF CYTOSINE DEAMINASE AND
URACIL PHOSPHORIBOYSLTRANSFERASE FOR SUICIDE
GENE THERAPY
Runyan R University of Washington School of Medicine, Seattle, WA.
Purpose of Study: Suicide gene therapy (SGT) is a process of selectively
introducing genes into tumor cells causing them to express enzymatic activities, making them susceptible to specific antimetabolites. Administration
of a prodrug results in the formation of the toxic antimetabolite in tumor
cells. Cytosine deaminase (CD), a promising prodrug gene therapy enzyme,
converts cytosine to uracil. CD also converts 5-fluorocytosine (5FC) to 5fluorouracil, which is a potent antimetabolite that inhibits DNA synthesis
and RNA function. Uracil Phosphoribosyltransferase (UPRT) is also a key
enzyme in the conversion of 5FC to the toxic metabolite. Both the yeast and
E. coli CD and UPRT genes have been utilized in SGT. A major problems
associated with this suicide gene/prodrug combination is a low affinity displayed by the gene product for the prodrug. One way to overcome the limited
production of 5FdUMP is by the fusion of key enzymes in the prodrug
activation pathway. Fusing the two enzymes responsible for the initial and
secondary limiting steps in prodrug activation has shown substantial improvement in tumor reduction.
Methods Used: Using the pETHT vector as a backbone, two novel combinations of CD and UPRT were constructed: bCD/yUPRT and yCD/bUPRT.
These new constructs were then transformed into the E.coli strain GIA39
(DE3) and plated, after which plasmid DNA was isolated from single colonies. Restriction digestion and genetic sequencing were performed to confirm
successful cloning. Genetic complementation studies tested protein synthesis
and function using E.coli stains that are deficient in UPRT and CD.
Summary of Results: Sequencing confirmed successful genetic cloning
of bCD/yUPRT and yCD/bUPRt. Through genetic complementation studies
we show that both constructs exhibit increased CD and UPRT enzymatic
activity, when compared to the control bCD/bUPRT and yCD/yUPRT constructs. These novel gene constructs have also demonstrated increased stability and productivity.
Conclusions: These experiments will ascertain the best possible combination of yeast and bacterial CD and UPRT fusions. A suicide gene therapy
with greater sensitivity to the prodrug will lead to a faster and more potent
way to suppress tumor growth with reduced side effects.
Western Student Medical Research Forum
Student Session V Y Surgery
1:30 PM
Friday, January 28, 2011
321
IMPROVED METHOD FOR CHRONIC CANNULATION
OF THE RAT THORACIC DUCT FOR MULTIPLE POOL
KINETIC STUDIES
Douglas MJ, Flake N, Witte M University of Arizona College of Medicine,
Tucson, AZ.
Purpose of Study: Few studies of pharmaco/metabolokinetics incorporate
lymph sampling for more accurate and insightful multiple pool analysis and
translational implications. This deficiency is in part due to relative inaccessibility of the lymphatic compared to blood vasculature and the difficulties
in initial cannulation and longer term lymph collection/drainage experiments
particularly in small animals.
Methods Used: To surmount these challenges, we refined techniques and
performed pilot multiple pool sampling experiments in 6 male Wistar-fuzzy/
Sprague Dawley rats weighing È350 gm. Following a corn oil meal to visualize milky intestinal lymphatics and ketamine-xylazine anesthesia, the
abdominal cavity was entered, and under a Wick dissecting microscope, the
cisterna chyli dissected up to the thoracic duct. The thoracic duct was then
ligated and a .012id x.240od silicone tube passed into the cisterna chyli,
secured with tissue glue, and serial timed samples of lymph collected. Simultaneous samples of central venous blood/serum were obtained by cannulation of the external jugular vein. Lymph drainage was maintained for
a period of up to 6 hours at which time collections were terminated.
177
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Cisterna chyli cannulation and drainage was per-
Summary of Results: 23,418 patients were identified. 1,643 (7.02%)
formed successfully and the lymph/blood sampling protocol carried out effectively. Milky lymph flow at equilibrium averaged 454 T 246 microliters/hr
(mean T SD), but declined gradually over the 6-hour collection period.
Conclusions: This rat preparation should provide the basis for further dynamic experiments examining multiple pool kinetics of absorption, distribution and fate of a wide variety of drugs, toxins, microrganisms, and
trafficking cells. Such experiments would contribute to improved fundamental understanding of the action and effectiveness of these agents, fresh
insights into related processes under normal and pathologic conditions, and
potential treatment approaches.
patients developed nonunion and 356 (1.52%) patients developed malunion
following treatment. Open treatment with internal fixation yielded the highest
incidence of nonunion (16.8%). Incidence of nonunion following external
fixation was 13.6%, and incidence of nonunion following intramedullary
(IM) nailing was 14.24%. 22 patients (4.2%) of the external fixation group
and 158 patients (2.6%) of the IM nailing group developed malunion. Of all
males undergoing external fixation, 39 (10.83%) suffered nonunion, whereas
33 (19.4%) of all females undergoing external fixation suffered nonunion
(p=0.0079). 122 (1.8%) of males and 62 (1.3%) of females in the closed
treatment without manipulation group developed nonunion (p=0.0412).
Conclusions: Our study demonstrated a significant difference in gender
with respect to nonunion incidence in two of the five treatment modalities:
external fixation group and closed treatment without manipulation. Our
study also generated a signifiant difference in gender with respect to malunion incidence in two of the five treatment modalities: open treatment with
internal fixation and closed treatment without manipulation. Further studies
should examine this gender disparity.
322
THE INCIDENCE OF USE OF PLATELET RICH PLASMA
IN ORTHOPEDIC SURGERY IN IDAHO AND WASHINGTON
Butikofer SA1,2, Seegmiller J2 1University of Washington, Seattle, WA and
2
University of Idaho, Moscow, ID.
Purpose of Study: To measure the frequency of platelet rich plasma (PRP)
use in orthopedic surgery in Idaho and Washington, and to report the factors
that influence orthopedic surgeons to use or not use PRP.
Methods Used: A questionnaire was developed to gather data in three areas:
physician knowledge of PRP applications, PRP use in orthopedic practices,
and factors influencing PRP use. 45 orthopedic physicians in Idaho and
Washington responded to the questionnaire. Analysis of variance was used to
determine if any significant differences existed in PRP use when influential
factors, such as high or low PRP knowledge, were compared. In addition,
correlation tests were run to identify any relationships between use and potential influencing factors.
Summary of Results: Respondents described their own familiarity with
PRP in orthopedic surgery as more than Fsomewhat familiar_ on a four
point scale (mean=3.27 T 0.72). Our sample used PRP less than Frarely_
(mean=1.89 T 1.02) and a Pearson_s correlation test found that physician
knowledge of PRP and PRP use was weakly correlated (r=.235). Physician
knowledge of PRP, experience using PRP, and improved patient outcomes
were rated on a four point scale as the most influential factors for physicians
who use PRP with means of 3.48 T 0.59, 3.16 T 0.87, and 3.22 T 0.95,
respectively. For physicians who choose not to use PRP, the most highly
rated factor that influenced this decision was a lack of randomized controlled
trials involving platelet rich plasma use in orthopedic surgeries with a mean
of 3.54 T 0.74.
Conclusions: Orthopedists are generally knowledgeable about PRP in orthopedic applications. Despite high levels of knowledge and familiarity with
PRP, 46% of our respondents do not use PRP in their practices. Knowledge,
training, and experience have little effect on whether physicians choose to use
PRP or not. However, knowledge, experience, and positive patient outcomes
are influential to physicians who do choose to use PRP. Whether the physician
used PRP or not, all seemed to recognize a lack of literature evidence for
some applications and considered this an important influencing factor.
323
GENDER DIFFERENCES IN NONUNION AND MALUNION
FOLLOWING TIBIAL SHAFT FRACTURES: A REVIEW
OF 5 TREATMENT MODALITIES
Chapman LW, Hame S, Wang JC David Geffen School of Medicine at
UCLA, Los Angeles, CA.
Purpose of Study: Tibial shaft fractures are the most common type of
tibia fracture and predominately occur following car accidents, sports injuries, and falls. The purpose of this study was to compare 5 treatment modalities for tibial shaft fractures and to evaluate the potential benefit of
undergoing a specific treatment given gender. Potential benefit in this study
involves minimizing the incidence of nonunion and malunion following
treatment.
Methods Used: Patients undergoing treatment for tibial shaft fractures
from years 2004Y2009 were retrospectively reviewed using a commercially
available online database of private insurance billing records. Fracture
treatment was identified by CPT code. The 5 treatment modalities were external fixation, intramedullary nailing, open treatment with internal fixation, closed treatment without manipulation/reduction, and closed treatment
with manipulation/reduction. Records were cross referenced for nonunion
and malunion.
178
324
POOR CORRELATION BETWEEN PRE-TRANSPLANT
VISUALIZED ESTIMATE OF DECEASED DONOR LIVER
STEATOSIS AND REPERFUSION BIOPSY ASSESSMENT
Fuller SD, Mangus S, Fridell J, Vianna R, Milgrom M, Tector J Indiana
University School of Medicine, Indianapolis, IN.
Purpose of Study: Pre-transplant deceased donor liver steatosis is estimated by the procuring surgeon by gross appearance of the organ, by a frozen
section biopsy, or a combination of the two. This estimate is often the determining factor for the use or non-use of the organ. Permanent section
biopsy of the reperfused liver may give a more accurate and reliable assessment of steatosis. This paper assesses the correlation between the
surgeons_ estimate of hepatic steatosis and the steatosis measured on permanent section biopsy from the reperfused liver.
Methods Used: This study is a retrospective review of all deceased donor
liver transplants at a single center over 5 years_ time. The surgeons_ estimates
for all procured livers were recorded from the original on-site records from
the organ procurement organization. There was no standardization to the
reporting such that each recorded value reflected the estimate recorded by
the on-site coordinator as stated by the procuring surgeon. All reperfusion
biopsies were read by experienced liver transplant pathologists at a single
center. A scatter plot with best fit line was utilized to determine correlation.
Summary of Results: Data were available for 485 liver transplants. Correlation between the surgeon estimated steatosis and assessment by permanent biopsy was r2=0.26. The surgeon underestimated steatosis in 25 cases
(5%), overestimated in 110 cases (23%), and was in agreement (+/j 5%) in
350 cases (72%).
Conclusions: These results suggest a reasonable ability of the procuring
surgeon to determine the presence of liver graft steatosis on site at the time
of organ procurement, but the estimate of the extent of steatosis correlates
poorly with that seen on permanent reperfusion biopsy.
325
VARIATIONS IN THE QUALITY OF CARE DELIVERED
TO PATIENTS UNDERGOING TOTAL HIP REPLACEMENT
AT THREE AFFILIATED HOSPITALS
Gurbani A1, SooHoo N2 1UCLA David Geffen School of Medicine,
Los Angeles, CA and 2UCLA David Geffen School of Medicine, Los
Angeles, CA.
Purpose of Study: While excellent clinical results have been seen with
total hip replacement, there has been extensive documentation of variations
in outcomes due to factors such as hospital and surgeon volume. These
disparities raise concern towards possible variations in the quality of care
being delivered. This study attempts to identify such variations at three affiliated hospitals by measuring adherence to an explicit set of evidence-based
quality measures.
Methods Used: Retrospective chart review was used to examine the
quality of care delivered to all patients receiving total hip replacement at the
Santa Monica-UCLA Medical Center (SM-UCLA), a private community hospital, in 2009. Both electronic and paper charts were examined, including
physician notes, ancillary service notes, results, and orders. Quality of care was
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
measured by determining adherence to a set of 32 evidence-based quality
indicators that were created using the RAND/UCLA Modified-Delphi expert
panel methodology.
Summary of Results: The results given represent a small initial sample
of thirteen patients (of a total of two-hundred and seven). The overall rate
of adherence for this sample was 56% (95% CI 50Y62%). There was significant variation between individual quality indicators. For example, preoperative risk assessment was conducted in all cases in this sample (100%),
while a complete physical examination was rarely documented (8%).
Conclusions: The initial results indicate an overall poor rate of adherence
to quality measures in this small sample. Specifically, adherence rates were
lower for preoperative and postoperative indicators than for intraoperative
indicators. We are in the process of collecting and analyzing data for the
remainder of patients from SM-UCLA. Concurrently, collaborators at the
UCLA Medical center, a high-volume academic center, and Harbor-UCLA
Medical Center, a lower volume public hospital, are undertaking this process. Once completed, we will conduct statistical analyses to determine
whether there is a significant difference in adherence to these measures
amongst these three sites. The finding of variations in care may be key
in identifying correctable causes for variable outcomes of total joint
replacement.
326
USE OF NEGATIVE PRESSURE THERAPY ON CLOSED
SURGICAL INCISIONS
Huberty SA1, Gabriel A2, Gupta S2 1University of Washington, Vancouver,
WA and 2Loma Linda University Medical Center, Loma Linda, CA.
Purpose of Study: Management of complicated wounds pose a continual
challenge to health care providers, particularly in patients with associated
risk factors and co-morbidities. Negative pressure wound therapy (NPWT)
is becoming the standard of care in open wounds. This study evaluated
the role of NPWT in closed incisions in patients with high risk factors for
wound breakdown.
Methods Used: 15 consecutive patients were evaluated in this pilot study.
VAC therapy (KCI) was utilized as our NPWT of choice and was applied
in the operating room to clean, closed surgical wounds. GranuFoam Silver
dressing (KCI) was applied to all closed wounds. The therapy was initiated
at 125mmHG below ambient pressure in a continuous mode for 3Y5 days.
Inclusion criteria included a minimum of one of the risk factors: BMI 935,
Diabetes Mellitus (DM), Peripheral Vascular Disease (PVD), smoking, history of radiation, steroid use, and malnutrition measured by albumin and
prealbumin. Mean follow-up was 9 months.
Summary of Results: All patients completed their treatment successfully.
Surgical incisions ranged from abdominal, chest and lower extremity incisions. There were no returns to the operating rooms and no wound breakdowns. No other complications were seen in our pilot study.
Conclusions: These results are suggestive of NPWT role over incisional use
in high-risk patients. Clearly randomized trials need to address this in the
near future; however, we recommend its use on the high-risk patients who
are undergoing elective or emergent procedures to minimize wound breakdown and improve healing.
327
INCIDENCE AND CORRELATES OF FAILURE FOR
VENTRICULOPERITONEAL AND LUMBOPERITONEAL
SHUNTS IN PATIENTS WITH IDIOPATHIC INTRACRANIAL
HYPERTENSION
Martin RL1,2, Lambert W2, Tanne E2, Choi D2, Lobb B2, Loboy S2
1
University of Washington School of Medicine, Seattle, WA and 2Oregon
Health and Science University, Portland, OR.
Purpose of Study: Idiopathic Intracranial Hypertension (IIH) is a condition of raised intracranial pressure with unknown cause. Symptoms include
headache, pulsatile tinnitus, stiff neck and radicular pain, transient visual
obscurations due to papilledema, and in severe cases, visual loss and blindness. Although IIH can occur in people of every demographic group, studies
have found that gender, age, and obesity are strong risk factors. For example,
men have an estimated incidence of 1.5 cases of IIH per 100,000, while
overweight women of childbearing age (15Y44) have a significantly higher
incidence at 20 cases per 100,000. With the number of overweight and obese
women on the rise, the health burden of IIH is expected to increase. One of
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
the primary surgical options for IIH patients includes cerebral spinal fluid
(CSF) shunt procedures. Two major types of shunts are used to treat IIH:
lumboperitoneal (LPS) and ventriculoperitoneal (VPS). While successful
at providing acute symptom relief, LPS and VPS frequently require one
or more revisions and have a number of complications. Current information concerning the comparative incidence of failure in LPS and VPS is
inconclusive.
Methods Used: Using the largest IIH patient database in the world, our
study will be able to assess the comparative incidence of failure between
LPS and VPS in a large cohort of geographically diverse patients. Using life
tables analysis and Chi-square test for difference of two proportions, we
will determine whether life table estimated failure rates differ between LPS
and VPS at 3, 6, 12, and 24 months. Additionally, we will determine whether
BMI, lumbar opening pressures, and time from initial diagnosis to first surgery, are associated with failure.
Summary of Results: Pending: will be available upon presentation.
Conclusions: This information will provide patients and providers with
critically important evidence needed to make informed treatment decisions.
328
A CASE OF MIXED GONADAL DYSGENESIS WITH
45,X/46,XRINGY/46,XY KARYOTYPE AND ITS
LAPAROSCOPIC MANAGEMENT
Molavi I1,2, Masterson J1,3 1University of British Columbia, Vancouver, BC,
Canada; 2Office of Pediatric Surgical Evaluation and Innovation, Vancouver,
BC, Canada and 3BC Children’s Hospital, Vancouver, BC, Canada.
Case Report: This is a case report on a 13 month old boy who presented
with undescended testes and was found to have Mixed Gonadal Dysgenesis
with karyotype 45,X/46,XringY/46,XY. MRI investigations revealed a left
intra-abdominal gonad in the iliac fossa, whereas no contralateral gonadal
structure was identified. Imaging suggested the presence of a cervix with a
small uterine structure at the superior aspect. Laproscopic exploration documented a hemi-uterus with a fallopian tube and streak gonad on the right
side. On the left side, a testicle was located in the abdomen proximal to the
internal ring of the inguinal canal. The testicular vessels were double clipped
for a stage I Fowler-Stephens orchidopexy procedure, and no other abnormalities were identified. It is essential to emphasize the importance of current
diagnostic tools in treating and managing patients with sexual development
disorders such as Mixed Gonadal Dysgenesis. Laparoscopic management not
only provides minimally invasive surgery, but it also facilitates all necessary
procedures including evaluation, biopsy, and gonadectomy for diagnosis and
treatment.
329
ALLOGRAFT AND MICROSCOPE USE IN SINGLE-LEVEL
ANTERIOR CERVICAL DISCECTOMY AND FUSION
SURGERY DO NOT INCREASE THE INCIDENCE OF
SERIOUS INFECTION
Pannell W1, Ngo S3, Wang C2, Ellerman J2 1UCLA, Los Angeles, CA;
2
UCLA, Los Angeles, CA and 3UCLA, Los Angeles, CA.
Purpose of Study: Wound infection requiring irrigation and debridement
(I&D) is a rare complication of anterior cervical discectomy and fusion
(ACDF) surgery but not an unimportant one. The purpose of this study was
to determine if graft choice or microscope use during surgery affected the
incidence that patients had to return to the operating room for I&D.
Methods Used: A retrospective review of patients who have undergone
single-level ACDF surgery was conducted using a commercially available
database of insurance billing records. CPT codes were then used to identify
patients based on graft choice and microscope use. These records were crossreferenced for CPT codes for regular or deep irrigation and debridement
(I&D) within 45 days of surgery.
Summary of Results: 24,298 patients underwent single-level ACDF surgery from 2004Y2009. Allograft was used in 13,656 (56.2%) of patients,
autograft in 1410 (5.80%) and microscope in 2341 (9.63%). Irrigation and
debridement was performed within 45 days in 1 of the microscope patients
(0.043%) and 10 of the non-microscope patients (0.046%). The difference
in complication rates between these two groups was not significant. Allograft
use was associated with a higher rate of I&D (0.051%) than autograft use
(0%). Within the allograft group, microscope use was associated with a higher
179
Western Regional Meeting Abstracts
Journal of Investigative Medicine
rate of I&D (0.084%) than was the non-microscope group (0.048%), however,
this association also did not reach significance.
Conclusions: While microscope use was not associated with a higher rate
of I&D by itself, when combined with allograft use it was associated with a
non-significant increase in the rate of I&D. Since even these small variations
can be useful in clinical practice, it is important to note that 7 of the 11 (64%)
reported I&D cases were from allograft patients. This may seem small, but it
should not be ignored because bringing the patient back into the operating
room is time consuming, costly, and increases the patient_s recovery time.
330
SURGICAL MANAGEMENT IN THE TREATMENT OF
FIBROUS DYSPLASIA
Richards SM, Adetayo OA, Salcedo SE, Ray AO, Gupta SC Loma Linda
University Medical Center, Loma Linda, CA.
Case Report: The patient is a 34 year old otherwise healthy male who was
diagnosed with fibrous dysplasia at age 16. He presented with complaints of
Preoperative CT illustrating fibrous dysplasia of the orbital region.
&
Volume 59, Number 1, January 2011
new onset frequent headaches and blurry vision in the left eye. The patient
underwent a left fronto-temporo-orbitozygomatic craniotomy, cranioplasty
and microsurgical decompression of the optic nerve. This was followed by
alloplastic reconstruction of the forehead and supraorbital rim as well as
orbital wall reconstruction. This was achieved using a Synthes PEEK implant
which was prefabricated based on the patient_s preoperative CT images. The
implant was made in two pieces to allow flexible contouring of the forehead
and orbital rim.
331
AN ATYPICAL PRESENTATION OF A PANCOLONIC
VENOUS MALFORMATION
Wong F, Blair G University of British Columbia, Vancouver, BC, Canada.
Purpose of Study: To report a unique case of gastrointestinal venous
vascular malformation.
Methods Used: The patient is a 13 month-old girl. Methods used to elucidate her medical situation include CBC & Peripheral Smear, Coagulation
Investigations, Ultrasound, Abdominal X-ray, CT angiogram, Laparotomy,
and Pathology and Histology of the surgical specimen.
Summary of Results: Venous vascular malformations (VM) are common
in infants in the head, neck, extremities, trunk and especially cutaneous
regions. The occurrence of a VM in deeper structures such as the colon,
however, is uncommon. The location of reported gastrointestinal venous
vascular malformation (GIVM) cases is usually in the sigmoid colon and
rectum. Almost all cases in literature presented with hematochezia. Herein we
describe a unique case of a pancolonic VM with atypical locationVbetween
the transverse colon and mid-sigmoid colonVand atypical presenting
symptomVan upper GI bleed. Lab results suggested a consumptive coagulopathy and imaging revealed the section of colon involved. The hypothesized pathogenesis of the presenting upper GI bleed is a Kasabach-Merritt
like syndrome caused by the VM. The venous malformation resulted in stasis
of blood that activated the coagulation cascade. This led to the formation of
blood clots and phleboliths and depleted clotting factors such as fibrinogen
and platelets. The thrombosed vessels caused injury to passing erythrocytes
resulting in anemia. The coagulopathy complicated the upper GI bleed. In
order to correct the source of the coagulopathy, the VM was resected. Pathology of the surgical specimen confirmed thrombi and phleboliths in venous channels as well as recanalized thrombosed vessels. The findings
confirmed the suspicion of coagulation within the malformation that may
have led to the consumptive coagulopathy.
Conclusions: In conclusion, we note that gastrointestinal venous vascular
malformations can present in an atypical way. The presenting symptom
may not be hematochezia. In this case, an upper GI bleed was the only
presenting symptom that hinted at the underlying coagulopathy and the rare
gastrointestinal venous malformation.
General Internal Medicine and Aging
Concurrent Session
8:30 AM
Saturday, January 29, 2011
332
PERIORAL WRINKLES ARE ASSOCIATED WITH GENDER,
AGING, AND SMOKING: DEVELOPMENT AND ANALYSIS
OF A GENDER-SPECIFIC PHOTONUMERIC SCALE
Alloplastic reconstruction of the forehead and supraorbital rim.
180
Cheng N, Chien A, Kang S Johns Hopkins University, Baltimore, MD.
Purpose of Study: Perioral rhytides, or wrinkles around the mouth, are a
highly prevalent phenomenon and common chief complaint among older
adults. Proposed etiologies include sun exposure, phototype, cigarette
smoking, and hormone replacement. Clinical impression suggests perioral
wrinkling affects females to a greater extent than males. Yet, no existing
grading scale accounts for this gender difference. To this end, we developed
an objective, gender-specific photonumeric scale for the assessment of
perioral rhytides and investigated possible associated factors.
Methods Used: A total of 143 individuals were enrolled (72 F, 71 M, aged
21 to 91). Standardized photographs were taken of each participant_s entire
face and perioral area. Five photographs were selected as standards and were
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
assigned grades of 0, 2, 4, 6, and 8, in ascending severity of perioral wrinkling. Subjects were evaluted by three blinded graders and also completed a
questionnaire of 24 potentially associated demographic and lifestyle factors.
Summary of Results: Perioral wrinkling was statistically significantly
correlated with females over age 45 (mean score 4.8 F, 2.9 M, p G 0.001) and
duration of smoking (mean score 3.4 smokers, 2.1 nonsmokers, p = 0.0005).
Other correlations included number of children birthed (p = 0.004), years of
recreational sun exposure (p = 0.0001), years of lip product use (p G 0.0001),
years of hormone replacement therapy (0.0001), and years of vitamin use or
other health supplementation (p G 0.0001). Analysis of the three graders_
maximum range of disagreement (MRD) showed good inter-rater reliability,
and repeat grading and MRD analysis two weeks later showed good intrarater reliability. A modified multiple linear regression model selected gender,
age, and smoking as the best predictors of severity of perioral wrinkling.
Conclusions: We developed a reliable nine-point photonumeric scale to
evaluate perioral wrinkling in female subjects. The correlation with smoking
augments our understanding of tobacco_s detrimental effects on aging.
Studies are ongoing to characterize perioral wrinkling in African-Americans
and investigate possible ethnicity-related patterns of aging.
333
THE EFFECTS OF CANCER DOSING OF BISPHOSPHONATES
ON THE MECHANICAL PROPERTIES OF BONE DEPEND
ON DURATION OF TREATMENT
Luo T, Allen M Indiana University School of Medicine, Indianapolis, IN.
Purpose of Study: Bisphosphonates (BPs) are a class of drugs used to treat
osteoporosis as well as bone metastases related to various cancers (breast,
prostate, multiple myeloma). They reduce bone loss via inhibition of osteoclasts. Our goal was to study side effects associated with long-term and
high dose bisphosphonate use, such as in cancer regimens. Previous animal
studies have shown that BP dosing for osteoporosis produce significant
changes in the mechanical properties of bone, including decreased toughness, the amount of energy that can be absorbed by bone before fracturing.
We hypothesized that cancer dosing regimens of zoledronate, an intravenous BP, reduce bone toughness, and that this effect would be dose- and
duration-dependent.
Methods Used: The model for the experiment was rib bones from skeletally mature beagle dogs, which were part of three separate experiments. The
first experiment tested monthly dosing of zoledronate for 3 and 6 months.
The second experiment assessed biweekly dosing for 3 months. The third
experiment examined biweekly dosing for 9 months with secondary treatment of dexamethasone. Each experiment had a set of untreated controls.
Histological analysis, peripheral quantitative computed tomography (pQCT),
and biomechanical testing were performed on all bones.
Summary of Results: Dosing of zoledronate either monthly or biweekly
reduced remodeling and increased mineralization of the bone but did not alter
mechanical properties at 3 or 6 months. In the 9-month study, we also found
reduced remodeling and increased mineralization, but in addition, the bones
showed significantly lower biomechanical properties in the zoledronate-treated
samples compared to the controls, especially in energy to failure (938 mJ to 497
mJ, p = 0.02) and toughness (41.64 MJ/m3 to 17.06 MJ/m3, p e 0.001).
Conclusions: These results indicate that the effects on bone toughness are
duration-dependent. Interestingly, in the animals treated for 9 months with
zoledronate plus dexamethasone, the dexamethasone-treatment attenuated
the effects of zoledronate and kept bone properties at control levels. Additional studies will be needed to understand how dexamethasone is altering
the effects of zoledronate on bone toughness.
334
CHARACTERIZATION OF ALPHA-CRYSTALLIN UPTAKE
INTO EYE LENS CELLS AS A NOVEL TREATMENT FOR
CATARACT AND RETINAL DEGENERATION
Christopher K, Mueller N, Petrash JM University of Colorado School of
Medicine, Aurora, CO.
Purpose of Study: Protein aggregation disorders are among the most
common diseases afflicting humans and tend to have the fewest treatments
available. In the eye, cataracts can be formed by aggregation of F-crystallin
protein subunits in the lens. Similarly, a mutation in the rhodopsin gene leads
to aggregation and blindness in patients with retinitis pigmentosa. A family
of small heat shock proteins called the >-crystallins show chaperone-like
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
activity (CLA) in the eye as well as elsewhere in the body. During cataract
formation, these proteins tend to accumulate at the membrane of lens cells and
lose their CLA function. Replacing the pool of >-crystallin proteins is a novel
therapeutic strategy to restore CLA in the eye and prevent protein aggregation.
Methods Used: Using human lens epithelial (HLEB3) cells, we characterized the cellular uptake of one of the >-crystallin proteins, >B-crystallin,
and monitored its activity once inside the cell. Recombinant >B-crystallin
was purified and labeled with a fluorescent probe to enable us to monitor
its real-time uptake and subcellular distribution.
Summary of Results: Uptake of wild-type >B-crystallin after packaging
into a cationic polymer transfection reagent proved most effective, however
the reagent caused reduced cell viability. Additional mechanisms to enhance
protein delivery are being studied such as our modified >B-crystallin fused
to a protein transduction domain. Stability of >B-crystallin once inside the
HLEB3 cells was also investigated. Using gel electrophoresis methods, we
observed that the protein was not being rapidly degraded by the cell. Additionally, a series of point mutants of >A-crystallin have been developed,
expressed, and purified. These mutants have the potential for increased
CLA and may be a more effective therapeutic option. Furthermore, a cell line
with inducible aggregation-prone rhodopsin expression was developed and is
being used to test the efficacy of treatment with our therapeutic >-crystallins
to prevent or decrease the amount of protein aggregation in the cell.
Conclusions: The >-crystallin family of small heat shock proteins remains
a promising approach to treatment of protein aggregation disorders in the
eye and elsewhere in the body.
335
PLASMA TESTOSTERONE IS ASSOCIATED WITH
FRAMINGHAM RISK SCORE
Chock BE1,2, Lin T4, Li C3, Swislocki A1,2 1University of California, Davis,
Sacramento, CA; 2VA Northern California Health Care System (VANCHCS),
Martinez, CA; 3University of California, Davis, Sacramento, CA and
4
University of California, Davis, Sacramento, CA.
Purpose of Study: The Framingham risk score predicts a patient_s 10 year
risk of developing cardiovascular disease. Many risk factors included in the
calculation of this score influence (or are influenced by) circulating testosterone. To further clarify the possible association between testosterone and
cardiovascular risk, as defined by the Framingham score, we analyzed data
from a VA database.
Methods Used: A retrospective chart review was performed. Inclusion
criteria were male sex and age 9 20. Exclusion criteria included pre-existing
cardiovascular disease, stroke, and diabetes. Testosterone supplements,
antiandrogen therapy, antihypertensives, or lipid-lowering medications were
not exclusion criteria, but were factored in data analysis. Laboratory and
clinical data were collected on veterans who had total plasma testosterone
checked in the year 2008. Framingham scores were calculated using an online tool from the National Heart Lung and Blood Institute. Using SAS v9.2,
robust linear regression assessed the association between total testosterone
and Framingham score. The Spearman rank correlation coefficient, Wilcoxon
rank-sum test, and Kruskal-Wallis test evaluated associations between total
testosterone and the other patient data. A p-value e 0.05 was considered
significant.
Summary of Results: 1,479 patients were included in the study. Mean age
was 61 years. There was a significant negative association between total
testosterone and Framingham score (estimate of regression parameter for
total testosterone j0.0042, p G 0.0001). Total testosterone was also positively
associated with HDL and negatively associated with total cholesterol and
triglycerides.
Conclusions: Lower plasma testosterone may suggest the presence of other
more traditional cardiovascular risk factors and potentially increased risk for
heart disease.
336
VITAMIN D INHIBITS PROLIFERATION AND INDUCES
DIFFERENTIATION OF C2C12 SKELETAL MYOBLAST
CELLS BY MODULATING THE EXPRESSION OF
PRO-MYOGENIC AND ANGIOGENIC GROWTH FACTORS
King KK1,2, Garcia LA1, Ferrini M1,2, Norris KC1,2, Artaza JN1,2
1
Charles R. Drew University of Medicine and Science, Los Angeles, CA
and 2David Geffen Medical School at UCLA, Los Angeles, CA.
181
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Purpose of Study: Skeletal muscle wasting is a serious public health
problem associated with aging, chronic disease, kidney dialysis and AIDS.
Vitamin D (VD) is widely recognized for its regulation of calcium and
phosphate homeostasis in relation to bone development and maintenance and
for its synergistic effects on target organs such as intestines, kidneys and PTH
glands. It has been shown to improve muscle performance and reduce falls
in VD deficient older adults. Little is known of the underlying mechanism
or role it plays in association with myogenic differentiation. In this study, we
examined the effect of 1,25-D -the active form of VD- at the molecular level
on myoblast cell proliferation, progression and differentiation into myotubes.
Methods Used: C2C12 myoblasts were treated with and without 1,25D in a
time course manner. VDR expression was analyzed by: Immunofluorescence
(IF), Real Time PCR (qRT-PCR) and Western Blots. Expression of muscle
lineage and proliferation markers was assessed by immunocytochemistry (ICC)
and western blots. Expression of pro-myogenic markers and angiogenic growth
factors was analyzed by qRT-PCR arrays and confirmed by qRT-PCR.
Summary of Results: Addition of 1,25D to C2C12 myoblast induces: a)
increase expression and nuclear localization of the VDR; b) decrease PCNA
(a cell proliferation marker) cell expression; c) increase expression of MyoD
and myogenin (early and intermediate myogenic markers respectively); d)
increase expression of IGF-2, FST (follistatin-a myostatin inhibitor), FGF1
(fibroblast growth factor 1, involved in muscle development and regeneration) and VEGF which has been demonstrated to promote myotube hypertrophy and increased myogenic differentiation; e) no changes in GDF11
expression; and f) decrease expression of IGF-1 and Mstn- the only known
negative regulator of muscle mass.
Conclusions: This study provides a mechanistic justification for VD replenishment in muscle waste conditions such as AIDS, cancer, congestive
heart failure, COPD, renal failure, and also in VD deficient older adults who
are known to have age-related loss of muscle mass and strength and an
increased rate of falls.
337
ALCOHOL USE DISORDERS IDENTIFICATION
TEST - CONSUMPTION QUESTIONNAIRE (AUDIT-C):
HOW OFTEN DO SCREENING RESULTS CONTRADICT
REPORTED DRINKING LEVELS?
Delaney KE1, Lapham GT2, Rubinsky AD2, Johnson ML2, Bradley KA2,1
1
Univ. of Washington, Seattle, WA and 2VA Puget Sound, Seattle, WA.
Purpose of Study: The AUDIT-C is a 3-item screening tool that asks about
quantity and frequency of alcohol consumption. Screening thresholds of Q4
(men) or Q3 (women) points have been found to balance sensitivity and
specificity for identifying alcohol use disorders and/or exceeding recommended limits (e14 drinks/week and e4 drinks/day for men; e7 d/wk and e3
d/day for women). However, some patients who report drinking within these
limits on the 3 questions can screen positive according to AUDIT-C score and
some who report exceeding limits can screen negative on the AUDIT-C.
Thus, AUDIT-C screening results (positive or negative) and reported drinking
on the AUDIT-C questions (above or below recommended limits) can be
discordant, though how often this occurs is unknown. The purpose of this
study was to evaluate the prevalence of AUDIT-C screening results that are
discordant with reported drinking among male and female drinkers in two
national samples: US general population and VA outpatients.
Methods Used: All combinations of responses to the 3 AUDIT-C questions
(Bresponse patterns[) were evaluated to identify screening results that were
discordant with reported level of drinking. Proportions of men and women
with discordant response patterns were estimated among drinkers in the US
(N=26,610) and VA (N=468,245) samples.
Summary of Results: For men, 14 AUDIT-C response patterns were discordant. Eleven responses patterns yielded negative screens but indicated
exceeding recommended limits and 3 response patterns yielded positive
screens but indicated drinking within limits. In the US and VA samples,
13.8% and 21.1% of male drinkers, respectively, had discordant response
patterns. For women, there were 7 discordant response patterns: 4 yielded
negative screens but indicated exceeding limits and 3 yielded positive screens
but indicated drinking within limits. In the US and VA samples, 10.6% and
12.1% of female drinkers, respectively, had discordant response patterns.
Conclusions: Many individuals who drink will have AUDIT-C screening
results that appear to contradict their reported alcohol consumption. Response patterns that yield these discordant results need to be considered when
brief alcohol interventions are provided.
182
338
AGE-RELATED CHANGES IN THE RETINAL NERVE FIBER
LAYER OF AFRICAN AMERICAN AND CAUCASIAN
HEALTHY SUBJECTS USING SPECTRAL DOMAIN OPTICAL
COHERENCE TOMOGRAPHY
Buono K1, Leite M1,2, Bowd C1, Weinreb R1, Medeiros F1,3, Zangwill L1
1
University of California San Diego, San Diego, CA; 2Federal University
of Sao Paulo, Sao Paulo, Brazil and 3University of Sao Paulo, Sao Paulo,
Brazil.
Purpose of Study: To evaluate the effect of aging and scan quality on
spectral domain optical coherence tomography (SDOCT) retinal nerve fiber
layer (RNFL) thickness in healthy participants of African descent (AD) and
European descent (ED).
Methods Used: AD (34 eyes) and ED (161 eyes) of 98 healthy participants
from the UCSD Diagnostic Innovations in Glaucoma Study (DIGS) and the
African Descent and Glaucoma Evaluation Study (ADAGES) underwent
RNFL imaging with Cirrus SDOCT (Carl Zeiss Meditec, Inc, Dublin, CA).
AD (30 eyes) and ED (120 eyes) of 75 healthy participants who underwent
RNFL imaging with Cirrus SDOCT also underwent RNFL imaging with
Spectralis SDOCT (Heidelberg Engineering, Inc, Heidelberg, Germany). Univariable and multivariable analysis of the effect of age, SDOCT quality score,
race, disc size and axial length on RNFL thickness was completed.
Summary of Results: There was significant (pG0.05) thinning with increasing age for the inferior Cirrus RNFL sector and the global, inferotemporal, and inferonasal Spectralis RNFL sectors. There was also significant
RNFL thinning with a poorer quality score for all of the Cirrus RNFL sectors
and for the temporal and superotemporal Spectralis RNFL sectors. The effect
of age and quality score was similar in AD and ED eyes and explained approximately 20% and 7%, respectively of the variation in the univariable
models of the inferior RNFL quadrant.
Conclusions: AD and ED eyes demonstrated age-related thinning of the
RNFL in the inferior quadrant, which is a location that is often affected early
in glaucoma disease progression. These results suggest that SDOCT shows
promise in providing quantitative data about the extent of age-related changes
in the RNFL of healthy eyes. Due to the observation that the SDOCT quality
score affects the RNFL measurement, future studies will need to account for
this relationship in order to ensure that measured RNFL changes in healthy
eyes are due to changes in age alone rather than changes in age-related SDOCT
quality scores.
339
DO DIABETES AND HYPERTENSION HAVE A SYNERGISTIC
EFFECT ON COGNITIVE FUNCTION IN MIDDLE AGE?
Chang J, Miller-Martinez D, Safaei Nili N, Crandall C, Karlamangla A
David Geffen School of Medicine at UCLA, Los Angeles, CA.
Purpose of Study: In the first half of this century, the number of individuals
over 60 years of age is expected to more than triple, representing the most
rapidly growing segment of the world population. Intact cognitive function is
essential for the well-being of older adultsVit determines engagement in the
workforce and social life.
Hypertension (HTN) and diabetes (DM) in midlife have each been associated with poorer cognitive functioning in later life. Few studies have
explored the effect of both diseases on cognitive function in middle age,
concurrently, and understanding these relationships may shed light on how
these common diseases alter cognitive function.
We used cross-sectional data from the second wave of a large populationbased study (the Midlife in the United States Study (MIDUS Study)) to assess
the impact of HTN and DM on the risk of cognitive function impairment in
middle-age (n=919), to test for synergistic effects between disease states,
and to examine changes in disease effect with age.
Methods Used: Multivariable linear regression (using SAS 9.2) was used
to evaluate the relationships between predictors (systolic blood pressure,
diastolic blood pressure, log(hemoglobinA1c), HTN, DM) and outcomes
(episodic memory, executive function), controlling for age, gender, education, and race.
Summary of Results: Adjusted for age, gender, education, and race, higher
systolic blood pressure was associated with slightly worse episodic memory (pG0.05) and executive function (pG0.01). Diabetic disease state was
associated with worse executive function (pG0.01), whereas hypertension
was not. No significant interactions were identified between continuous or
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
categorical variables representing diabetic and hypertensive status. There
was a significant age interaction: Log(hemoglobinA1c) was associated negatively with cognitive function only in older ages (965yo) (pG0.05).
Conclusions: Higher blood pressure and diabetic status are associated with
worse cognitive function, even in a middle-aged cohort. Poor glycemic
control (as measured by higher hemoglobinA1c) has a more pronounced
effect on cognitive function in older age, when vascular compensatory
mechanisms may not be as effective. No synergy was documented between
HTN and DM in their associations with cognitive function.
Hematology and Oncology II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
340
DEVELOPMENT OF AN INTEGRATED INTERDISCIPLINARY
ONCOLOGY ELECTIVE
Lai L, Ingledew P UBC Faculty of Medicine. Vancouver, BC, Canada.
Purpose of Study: A deficit in oncology education for medical students
exists throughout North America. Many medical programs expose students
to oncology sporadically, often in a discipline-specific manner which limits
the experience of the unique interdisciplinary nature of oncology. The goal
of this project is to develop an integrated interdisciplinary oncology clerkship, supplemented by online modules and virtual patients (VP).
Methods Used: The Kern approach to curriculum development was employed. A web-based survey was sent to 224 UBC medical students finishing
their first year of clerkship (3rd year of medicine), regarding adequacy of oncology training. The survey polled interest levels for an integrated oncology
clerkship, and student preference for mode of delivery for educational content. Following survey analysis, development of an online module covering
common types of cancer was initiated. In addition, a branching-logic Virtual
Patient (VP) case was developed using OpenLabyrinth (OL), a software tool
for authoring VP cases. Several software tools were evaluated for ability to
extract and analyze data from the OL database for purposes of education
research.
Summary of Results: The survey indicated a high interest in an integrated
oncology clerkship with an online module. 37% (82 of 224) of the target audience responded. 50% (41 of 82) of students had not interacted with cancer
patients. 70% (57 of 82) felt that an integrated oncology elective would be
useful. Nearly 80% (64 of 82) felt that online modules would enhance learning. A large proportion of students (62%, 51 of 82) rated their ability to discuss oncology issues with patients as poor to fair (the two lowest scores out of
a 5-point rating scale). To date, the learning modules for lung, prostate and
colorectal cancer have been written. A branching VP case for colorectal cancer
has been authored. Reports that analyze and display performance data derived
from students working through VP cases were created using Jasper iReports.
Remaining work includes development of a breast cancer module, and
authoring of more VP cases.
Conclusions: The gaps in oncology education can be met by the needsbased development of a new integrated oncology clerkship, especially if
supplemented by an online module. VP cases offer a unique learning environment, where students have the opportunity to practice clinical reasoning.
341
ACUTE MYELOID LEUKEMIA OUTCOMES FOLLOWING
BREAST CANCER TREATMENT
Ma CY1,2, Broady R1 1University of British Columbia, Vancouver, BC,
Canada and 2UBC, Van, BC, Canada.
Purpose of Study: Patients with breast cancer treated with chemotherapy
(alkylating agents/topoisomerase II inhibitors) and/or radiation are predisposed to therapy-related myelodysplastic syndrome or acute myeloid leukemia (tMDS/AML)[1]. Outcomes for tMDS/AML are poor compared to de
novo AML. Proposed causes are high frequency of unfavourable karyotypes,
prolonged cytopenias and injury to organs from therapy[1]. Investigation is
needed to determine factors that contribute to tMDS/AML development.
Methods Used: Retrospective chart review was done on 63 patients who
presented to the BMT clinic with tMDS/AML following previous breast
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
cancer treatment from 1985 to 2009. The median age at tMDS/AML diagnosis was 59 yrs (R=39Y84). The median latency between breast cancer and
tMDS/AML was 47 months (R=1Y447). Surgery only patients were excluded.
Summary of Results: After tMDS/AML diagnosis 21 patients had
palliative/supportive/no treatment and were excluded from further analysis.
45 patients received chemotherapy and 27 patients achieved remission. Of
these 9 patients received allogeneic bone marrow transplant and 4 had autologous transplant. 12 patients are alive without evidence of tMDS/AML or
breast cancer at time of study.
Patients were divided into 3 groups based on previous breast cancer
treatment: chemotherapy and radiation (C+R)(n=32), rad only (R)(n=23)
and chemo only (C)(n= 8). The median time between breast cancer and
tMDS/AML was longest for R (med=129 mos) vs. C+R and C (med=39;
31.5 mos). The median overall survival time between tMDS/AML and death
was 187 days (R=2Y2424).
Bone marrow karyotype was characterized as good (8 pts), intermediate
(29 pts) or poor (17 pts). The median overall survival for good, intermediate
and poor karyotype was 547, 212, and 167 days respectively.
Causes of death for patients were relapse of breast cancer (5 pts), cardiac
complications (3 pts), pulmonary failure (2 pts), sepsis (3 pts) and graftversus-host disease (3 pts with marrow transplant). The remainder of patients
died from a progression of tMDS/AML.
Conclusions: Our descriptive study provides support that cytogenetics and
treatments for cancer affects outcomes and overall survival of tMDS/AML.
References:
1. Chaplain G et al. Increased risk of acute leukemia after adjuvant
chemotherapy for breast cancer: a population-based study.
J Clin Oncol. 2000;18:2836Y2842.
342
TARGETING FOXM1 TRANSCRIPTION FACTOR IN
RHABDOMYOSARCOMA
Dolan JG1, Wan X2, Yeung C2, Helman LJ2 1University of Washington School
of Medicine, Seattle, WA and 2National Cancer Institute, Bethesda, MD.
Purpose of Study: Forkhead Box (Fox) M1 is a transcription factor involved with regulation of the G1/S, G2/M cell cycle checkpoints as well as
the mitotic spindle assembly checkpoint. FoxM1 has been reported in the
literature to be upregulated in different types of cancer. In a recent shRNA
screen, our lab showed that FoxM1 is necessary for growth and proliferation in rhabdomyosarcoma (RMS) cell lines. Recent studies have also
demonstrated that FoxM1 expression can be blocked using the thiazole
antibiotic Siomycin A. The purpose if this study was to determine the effects
of Siomycin A on FoxM1 in two human RMS cell lines, RD and Rh30.
Methods Used: To investigate our hypothesis, we used Western blot analysis and MTT proliferation assays. Western blots were prepared from cell
lysates of human RMS cell lines RD and Rh30 cultured in DMEM supplemented with FBS. Additional lysates from healthy donors and a variety of
other pediatric cancers were also used. Cells were cultured both in the
presence and absence of drug. Experiments were directed at reported targets
of FoxM1 involved with cell cycle progression as well as member of the Akt/
mTOR pathway.
Summary of Results: In this study, we show that FoxM1 is highly
expressed in multiple pediatric cancer cell lines, including RMS, when
compared to normal fibroblasts. Our results suggest that Siomycin A inhibits
FoxM1 and leads to decreased proliferation and increased cell death in a
dose dependant fashion. Interestingly, we found that in addition to blocking
FoxM1, Siomycin A inhibits the serine/threonine kinase Bub-1b. Bub-1b is
a mitotic spindle assembly checkpoint protein also found to be essential
for growth and survival of RMS cells in vitro and in vivo.
Conclusions: These data suggest FoxM1 and Bub-1b as potential therapeutic targets in rhabdomyosarcoma. Current studies are continuing to
characterize the relationship between these targets.
343
MELANOMA PATIENT INFORMATION: ASSESSMENT OF
THE QUALITY OF INTERNET-BASED INFORMATION
RESOURCES
Scali E1, Ingledew P2 1University of BC, Vancouver, BC, Canada and 2BC
Cancer Agency, Surrey, BC, Canada.
183
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Purpose of Study: Melanoma is the most serious form of skin cancer.
Many patients seek web-based information to guide their decision-making;
however, the internet is largely unregulated with few safeguards to regulate
its content. The purposes of this study were to apply a structured rating tool
to melanoma websites and to evaluate the quality of internet-based melanoma
information.
Methods Used: The study consisted of the creation of a standardized rating
tool and a structured review of the top 100 melanoma-related websites. A
rating tool was created based on a combination of existing methodologies.
It assessed websites based on affiliation, authorship, attribution, currency,
interactivity, readability, accuracy and coverage. The keyword Bmelanoma[
was input into three search engines. Exclusion criteria were applied and
the results were sorted to produce a list of the top 100 sites. The validity of
the tool was supported by assessing inter-rater reliability for a random subset
of sites.
Summary of Results: Websites were largely commercial in nature (48%,
n=48) with a smaller minority of non-profit sites (29%, n=29). Only half of
the sites clearly indicated the author (n=50) and the author_s credentials
(n=51) and 58% (n=58) indicated the author_s institutional affiliation. Less
than one-third identified the sources used to compile the information (n=31).
While 80% of sites (n=80) identified the date of creation of the entire site,
only 45% (n=45) had been updated within the past two years. The majority
of websites (87%, n=87) had interactive components; however, only 24%
(n=24) featured a discussion board and 9% (n=9) provided educational
support. The majority of websites (86%, n=86) provided information that
was accurate and 13% (n=13) were mostly accurate. 36% (n=36) published
information of treatment options by stage and only 15% (n=15) published
information on prognosis by stage.
Conclusions: Web-based melanoma information could be significantly
improved. This study revealed deficits in identifying authorship and attribution and less than half of the sites had been recently updated. While most
sites contained basic facts about melanoma, few contained accurate information based on disease stage. Information from this study can help to improve the quality of web-based information for melanoma patients.
344
LARGE SCALE CHEMOENZYMATIC SYNTHESIS OF THE
NON-HUMAN SIALIC ACID, N-GLYCOLYLNEURAMINIC ACID
Haidar Y, Pearce O, Varki A University of California, San Diego School of
Medicine, La Jolla, CA.
Purpose of Study: Classic and recent epidemiological studies continue to
reveal a link between human consumption of red meats/dairy products and
the incidence of carcinomas. While saturated fats or mutagens in grilled
meats are often blamed, these observations can also be explained by the
consumption of a non-human sialic acid, Neu5Gc, which is enriched in red
meats and milk. In addition, all humans have circulating anti-Neu5Gc
antibodies. The interaction of consumed Neu5Gc with these circulating
anti-Neu5Gc antibodies produces a chronic inflammatory state that could
have an important role in the progression of carcinomas. Major rate limiting
factors to further cellular and animal studies of the influence of Neu5Gc
on the incidence of carcinomas is the availability of Neu5Gc in large
quantities.
Methods Used: A novel method for the chemoenzymatic synthesis of
Neu5Gc was attempted (Figure 1). The synthetic scheme utilizes the bacterial enzyme, N-glycolylneuraminic acid lyase, which is purified by lysing
cells and running a manual Ni-sepharose high performance resin column.
The enzyme was covalently attached to an Amberzyme Oxirane resin which
simplifies purification and allows reutilization of the enzyme. Enzyme activity was checked via a thiobarbituric acid assay to determine optimal reaction conditions.
The cheap precursor, glucosamine, is converted to the synthetic intermediate, GlcNGc, by a simple chemical reaction. ManNGc, which is in
constant equilibrium with GlcNGc, is converted to the desired product,
Neu5Gc, via addition of the enzyme. After filtration of the resin, column
chromatography was run to purify the Neu5Gc.
Summary of Results: Large scale synthesis of Neu5Gc is achieved in two
simple reaction steps from the cheap precursor glucosamine and subsequent
purification by filtration and column chromatography.
Conclusions: The large-scale chemoenzymatic synthesis of Neu5Gc will
allow further studies of the link between human consumption of red meat/
dairy products containing Neu5Gc and the incidence of carcinomas.
184
FIGURE 1: Synthesis of Neu5Gc.
345
MITOCHONDRIAL DNA AND MINIMAL RESIDUAL DISEASE
ASSESSMENT IN PEDIATRIC ACUTE LYMPHOBLASTIC
LEUKEMIA
Villanueva N, Agsalda M, Chang L, Ernst T, Saito A, Pritchett L, Shiramizu
B University of Hawaii John A. Burns School of Medicine, Honolulu, HI.
Purpose of Study: Children treated for acute lymphoblastic leukemia
(ALL) have a risk for developing neurocognitive problems, which may be
associated with relapse. Relapse is contributed by minimal residual disease/or
persistent disease (MRD/PD) present in CSF or blood. MRD/PD-positive
specimens may have high mitochondrial DNA (mtDNA) copy numbers/cell,
a possible compensatory survival mechanism for a unique lymphoblastic
cellular phenotype in the brain. This study analyzes CSF for MRD/PD and
mtDNA copy numbers/cell in the subjects who are also undergoing special
MRI scans and neurocognitive tests.
Methods Used: CSF DNA from two consented newly diagnosed children
with ALL were assayed for MRD/PD status and mtDNA levels. Neurocognitive testing and special MRI scans (magnetic resonance spectroscopy,
diffusion tensor imaging, and high resolution MRI) without sedation were
also completed on these children and in age-matched non-ALL controls.
MRD/PD status was assessed by semi-nested real-time PCR using pooled
primer sets from the FR1/variable region of the IgH and two consensus 3_
primers. mtDNA copy numbers/cell were assessed using real-time PCR.
Summary of Results: Two children (001 and 002) were enrolled and had
CSF specimens available for MRD/PD and mtDNA copy number analyses.
Results summarized in Table 1. Subject 001 completed MRI scans and neurocognitive tests while Subject 002 tests are pending. Baseline neurocognitive test for subject 001 were normal while MRI scans indicated presence of
mild to moderate atrophy throughout the brain. Four control children have
also completed MRI scans and neurocognitive tests with normal results.
Conclusions: Although the results are too early to provide any conclusions
or statistical comparisons, MRD and mtDNA analyses of the CSF specimens
demonstrate the feasibility to perform the assays on CSF cells. Additional
children with ALL are continuing to be recruited.
TABLE 1. Summary of MRD and mtDNA Analyses of CSF
Equiv: Equivocal for MRD; Neg: Negative for MRD; Pos: Positive for
MRD; VH1: VH1 family; VH3: VH3 family; *mitochondrial DNA
copy number per cell; N/A: no specimen available.
346
DYNAMIC CONTRAST-ENHANCED MRI FOR PREDICTION
OF PATHOLOGIC RESPONSE TO NEOADJUVANT
CHEMOTHERAPY IN BREAST CANCER PATIENTS
Patel AP1, Chang D1, Chen J1,2, Lin M1, Mehta RS1, Su M1, Nalcioglu O1
1
University of California, Irvine, Irvine, CA and 2China Medical University
Hospital, Taichung, Taiwan.
Purpose of Study: To determine if changes in dynamic contrast-enhanced
(DCE) MRI pharmacokinetic parameters can be used to predict which
patients achieve pathologic complete response (pCR).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Methods Used: 57 breast cancer patients (mean age, 49 T 10 yrs) received
neoadjuvant chemotherapy (NAC) and underwent DCE-MRIs. 26 patients
were HER-2 positive and 31 patients were HER-2 negative. All patients
received doxorubicin and cyclophosphamide (AC) followed by a taxane
regimen. HER-2 positive patients received trastuzumab with taxane. DCEMRIs were obtained before NAC and at first follow-up, which was after 2
cycles of AC. For each MRI study, the whole tumor region of interest (ROI)
was drawn manually, slice-by-slice. A computer algorithm selected and labeled the hotspot that showed the greatest percentage enhancement within
each ROI. Enhancement kinetics from the whole tumor and hotspot were
fitted with the Tofts model to obtain the transfer constant (Ktrans) and rate
constant (kep). Pathology samples were categorized as pCR or non-pCR. The
ability of pharmacokinetic parameters to predict pCR was analyzed using
paired, two-tailed t tests. An artificial neural network was also used to select a
classifier based on Ktrans and kep parameters that could predict pCR. Receiver operating characteristic (ROC) analysis was performed and the area
under the ROC curve (AUROC) was examined.
Summary of Results: 30 patients (30/57, 53%) achieved pCR, including
19 HER-2 positive (19/26, 73%) and 11 HER-2 negative (11/31, 35%). In the
pCR group, the hotspot Ktrans value at the first follow-up was lower compared to the pre-treatment value (pG0.05). There was no significant difference
in Ktrans or kep between the pCR and non-pCR group. The AUROC to
differentiate between pCR and non-pCR was 0.76.
Conclusions: With a 53% pCR rate the NAC regimen used was effective,
especially in HER-2 positive patients (73%) who received trastuzumab with
taxane. While DCE-MRI pharmacokinetic parameters may not strongly
predict pCR after patients received only 2 cycles of AC, further analysis after
the administration of trastuzumab with the taxane regimen may demonstrate
a higher accuracy in prediction because pCR is most likely achieved using
targeted therapy with trastuzumab.
347
WHOLE ORGAN SEGMENTATION VERSE CYLINDRICAL
REGION OF INTEREST IN MEASURING CT RADIATION
DOSE OF ABDOMINAL ORGANS
Sandberg JK, Khatonabadi M, McNitt-Gray M David Geffen School of
Medicine at UCLA, Los Angeles, CA.
Purpose of Study: To determine whether accurate measurements of CT
radiation dose of abdominal organs can be estimated using cylindrical regions
of interests (ROIs) instead of whole organ dosage evaluations. This would
allow large dataset dose measurements and the development of organ dose
maps based on patient specific parameters. The end goal being, to track
lifetime patient radiation dose from CT.
Methods Used: Using a program developed by the lab, the livers, spleens
and kidneys of 19 female and 15 male contrast enhanced CT scans were
manually segmented. Cylindrical ROIs (2cm diameter/1.5cm height for
spleen/liver; 1cm diameter/1.5cm height for kidneys) were also taken at the
approximate volumetric centroid. A Monte-Carlo N-Particle (MCNPX)
simulation, using both fixed tube current (FTC) and tube current modulation
(TCM), calculated organ dose.
Summary of Results: Table 1 and 2 shows dosage RMSE and average
percent difference as well as the relation between patient abdominal circumference to organ dose for FTC and TCM respectively. As shown, full
organ and ROI organ dose measurements are comparable, organ dose is
dependent on patient size and organ dose does not differ across abdominal
organs.
Conclusions: Manual organ segmentation and its dosage calculations are
time consuming (È 5h / case), costly, challenging and require anatomical
expertise. The development of pre-calculated organ dose measurements using
accurate organ dose models would minimize the complications seen in
manual measurements. Thus, lifetime tracking of patient radiation dose
would be feasible, allowing investigators and clinicians to assess risk from
medical procedures.
Table 1: Fixed Tube Current (FTC)
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Table 2: Tube Current Modulation (TCM)
348
ORTHOVOLTAGE ROTATIONAL EXTERNAL BEAM
RADIATION THERAPY ON A BREAST CT PLATFORM:
A SIMULATION STUDY
Prionas ND, McKenney SE, Boone JM University of California Davis,
Sacramento, CA.
Purpose of Study: To evaluate the feasibility of a dedicated breast computed tomography (bCT) platform to deliver orthovoltage rotational external
beam radiation therapy through clinically relevant dose distributions, specifically partial breast irradiation, whole breast irradiation, and dose painting.
Methods Used: Monte Carlo simulations, using MCNPX 2.6.0, were based
on the geometry of a prototype bCT platform. A 178 keV monoenergetic
photon source was used to simulate a 320 kVp spectrum filtered by 4 mm of
copper, as validated by their depth-dose characteristics in polyethylene. The
source was rotated around a 14 cm voxelized polyethylene disk (0.1 cm tall)
or cylinder (9 cm tall) to simulate primary and primary + scattered photon
interactions, respectively. Beam collimation was varied in the x-y plane of
rotation (1Y14 cm) and in the z-direction (0.1Y10 cm). Dose deposition was
measured in the voxelized phantoms as mGy per incident photon.
Summary of Results: Varying collimation in x-y generated a family of 2D
dose profiles; dose distributions showed a cupped profile with high edge dose
using an uncollimated (14 cm) beam and increasingly peaked central dose
with low edge dose as collimation narrowed. The ratio of central dose to
cylinder edge dose was 14.8 for 1 cm and decreased to 0.6 for 14 cm collimation. Dose painting for multiple foci, a line distribution, and a ring distribution was demonstrated using multiple rotations with varying collimation.
A homogeneous dose distribution (G5% fluctuation) with skin sparing was
demonstrated using a weighted sum of four dose profiles. The mean scatter
to primary ratio across the central axial section of the cylinder ranged
from 0.007 (0.1 cm z-collimation) to 0.619 (10 cm z-collimation). Using
2 cm z-collimation, scatter tails decreased exponentially towards the cylinder
top/bottom to 3% of maximum dose deposition.
Conclusions: A bCT system is a feasible platform to deliver orthovoltage
rotational external beam radiation therapy for the treatment of breast cancer.
A variety of dose distributions can be generated that allow for partial breast
irradiation to a single focus, dose painting, and whole breast irradiation with
skin sparing. Dedicated bCT may serve as a platform for both diagnostic
breast imaging and radiation therapy.
349
PERSISTENT FEVER, SPLENOMEGALY, AND ANEMIA IN
A 2 YEAR OLD: A CASE OF SLOWLY PROGRESSING
HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS
Bowser AM1, Shliakhtsitsava K2, Arse E3 1UCSF Fresno, Fresno, CA;
2
UCSF Fresno, Fresno, CA and 3UCSF Fresno, Fresno, CA.
Case Report: Hemophagocytic Lymphohistiocytosis (HLH) is a rare disorder of cytokine dysfunction which may be familial, or associated with
infection, malignancy, or autoimmune disorders. Diagnosis is made by molecular analysis or by clinically meeting 5 major criteria (fever, splenomegaly,
cytopenia, hypertriglyceridemia and/or hypofibrinogenemia, and hemophagocytosis). Additional criteria include low or absent natural killer cell activity,
ferritin9500 mcg/L, or soluble CD2592400 U/ml. We report a 2 year old with
fever of unknown origin, massive splenomegaly, and persistent anemia, who
early-on did not meet HLH criteria. Given a high suspicion for HLH, the child
received an extensive workup, which revealed a diagnosis of HLH.
A 2 year old presented to the ER with 1 week history of fever, splenomegaly, and anemia (hemoglobin 5.5). Inpatient workup revealed evidence of
past EBV infection. Patient received 2 blood transfusions and was discharged
home. Patient returned to ER 2 weeks later with persistent fever and hemoglobin 4.8. Patient was readmitted for a workup, with HLH test results
summarized as follows:
185
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Day 1: triglycerides (144), fibrinogen (268), ferritin (346), bone marrow
biopsy (normal).
Day 8: triglycerides (143), fibrinogen (278), ferritin (462), NK-cell activity
(6), Soluble CD25 (5280).
Day 22: triglycerides (143), fibrinogen (152), ferritin (610), repeat bone
marrow biopsy (normal), lymph node biopsy (+hemophagocytosis), perforin
1 gene mutation analysis (indeterminate).
Patient remained nontoxic, despite having intermittent fevers, and needing regular transfusions. On a Day 22, cyclosporine and dexamethasone were
started per HLH protocol. Patient responded well, was discharged home on
Day 30. Patient is being followed in rheumatology clinic, and is currently in
clinical remission.
We propose that, in a child with persistent fever, splenomegaly, and
anemia, HLH should remain high on the differential. Workup for this rare
condition should be initiated even if the child does not meet diagnostic
criteria initially. We were unable to determine if our case was secondary to a
benign polymorphism or disease-causing mutation; additional mutation
analysis is ongoing.
Immunology and Rheumatology II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
350
ANALYSIS OF TEMPORAL PROTEIN SECRETION FROM
INDIVIDUAL T LYMPHOCYTES
Owens GE1, Ma C2,3, Cheung A2,3, Heath J2,3 1UCLA, Los Angeles, CA;
2
California Institute of Technology, Pasadena, CA and 3California Institute
of Technology, Pasadena, CA.
Purpose of Study: T cell functional diversity is critical for a robust immune
response to infection. This study aimed to develop a novel microfluidic platform
to analyze protein secretion by individual T cells at multiple time points.
Methods Used: An integrated microfluidic chip was assembled by layering
a PDMS slab containing a microfluidic network onto a glass slide containing
a DNA barcode microarray. Microfluidic networks were designed using
AutoCad and fabricated using two-layer soft lithography. DNA microarrays
were prepared using flow patterning of unique ssDNA molecules; addition of
complementary DNA-antibody conjugates created an antibody microarray on
the glass substrate. Each chip contained 768 microchambers. Fluid flow was
controlled with integrated pneumatic valves. The microchambers were loaded
with single or small numbers of Jurkat T cells stimulated with PMA/LPS.
After 24 hrs, detection antibodies tagged with biotin, and fluorescent probe
solution (Cy5-labeled streptavidin and Cy3-labeled M_ ssDNA) were passed
through the device sequentially. Fluorescence intensities of proteins from
each microchamber were quantified using the GenePix 4400B fluorescence
image analysis system.
Summary of Results: T cells were captured in individual micro-chambers.
Secretions of IL-2, TNF-alpha, granzyme B, and CCL-5 at 2, 12, and
24 hours were measured. Quantitative differences in secreted protein levels
were observed in single cells at different time points. Also, as the number
of cells within a chamber increased from single to ~100, CCL5 secretion
appeared to be upregulated.
Conclusions: Using a highly multiplexed microfluidic device, we captured
individual T cells and analyzed their protein secretions at different time
points. Future studies of T cell function both in both healthy volunteers and
patients with T cell dysfunction will permit us to assess functional heterogeneity on the single T cell level.
351
EPIGENETIC REGULATION OF B-1 B LYMPHOCYTES IN
LOW BIRTH WEIGHT INFANTS
Osborne A, McKnight R, Lane RH, Yost CC University of Utah, Salt Lake
City, UT.
Purpose of Study: Low birth weight (LBW) infants exhibit an increased
risk of infection, a condition that persists into adulthood. Specifically, LBW
adults demonstrate an attenuated immune response to T-cell independent
antigens. B-1 lymphocytes are the effector cells responsible for T cell independent antigen responses as well as early immune responses against
186
&
Volume 59, Number 1, January 2011
pathogens. B-1 lymphocytes are comprised of B-1a and B-1b cells. The
co-receptor CD5 is expressed on B-1a cells, but not on B-1b cells. The
expression of CD5 on the cell surface depends on the translation of two
alternative exons termed E1a and E1b. Transcripts initiated from the E1a
exon robustly express CD5 on the cell surface, while transcripts made
from E1b are retained in the cytoplasm. The alternative promoter for E1b
resides in a human endogenous retrovirus, which contains CpGs that can be
methylated. We hypothesize that LBW infants possess decreased numbers of
B-1b cells, and this will be associated with altered DNA CpG methylation of
the CD5 retroviral insert.
Methods Used: B cells were isolated from cord blood of LWB and normal
birth weight (NBW) infants. Cell populations were analyzed using flow
cytometry and the ratio of B-1b to B-1a cells was determined by counting
sorted cells. DNA from FACS-sorted B cells was isolated and subject to
bisulfite sequencing to determine the methylation patterns of the CD5 retroviral insert.
Summary of Results: Analysis has been completed on 40 patients. 3 were
LBW (birth weight of 10th percentile or lower). LBW patients have a decreased ratio of B-1b to B-1a cells compared to NBW patients (0.149 T 0.68 vs.
0.771 T 0.2745). B-1b cells from LBW infants also exhibit 30% and 33%
methylation of the CD5 retrovirus insert, compared to 45% and 50% methylation in NBW patients at two CpG sites within the CD5 retroviral insert.
Conclusions: LBW infants contain fewer B-1b cells and exhibit less
methylation within the CD5 retroviral insert. These are among the first
human findings that associate an epigenetic change within a gene directly
related to the targeted cell phenotype. We speculate that DNA CpG changes
of the CD5 retroviral insert will function as a marker for postnatal decreased
B-1b cell numbers.
352
CHART REVIEW ON 337 CASES OF KAWASAKI DISEASE
IN CENTRAL CALIFORNIA
Zhang Y1, Wright D2,1 1UCSF Fresno, Fresno, CA and 2Childrens’ Hospital
of Central California, Madera, CA.
Purpose of Study: Purpose of Study: Kawasaki disease (KD), a well
known pediatric vasculitis, is divided into 2 groups, typical or incomplete
KD, depending on how well the presentations satisfy the diagnostic criteria.
We were interested in understanding if there were any difference in terms of
outcome, lab values, and response to treatment between these two groups.
Methods Used: Methods Used: 337 cases seen at the Children_s Hospital
Central California (CHCC) from October 2000 to April 2009 who had KD as
one of their discharge diagnoses were reviewed. 199 cases which met the
criteria for KD or incomplete KD, had initial evaluation and were treated at
CHCC. Demographic data, initial presentation, initial and follow-up lab results, type of treatment, response to treatment, and outcome were reviewed
and compared between typical and incomplete KD.
Summary of Results: Summary of Results: In the 199 cases of KD
reviewed, there were no significant differences between the two groups in
terms of age of onset, gender, race, course of fever, season of onset, lab test
values, response to treatment, cardiac outcome, and non-cardiac complication
incidence. As for the initial clinical presentation, except for the course of
fever, typical KD presents with cervical lymphadenopathy, rash, peripheral
extremity change, oral mucosa change and conjunctival injection more than
the incomplete KD (all p=0.000).
Conclusions: Conclusions: Typical KD and incomplete KD most likely
share the same pathophysiological pathway, have no significant differences
in lab values, response to treatment or outcome. This reconfirms that the
diagnosis of incomplete KD should not change the clinical management
strategy.
353
MEASUREMENT OF THERAPY INDUCED NEUTRALIZING
ANTIBODIES TO INTERFERON (IFN)-A IN MULTIPLE
SCLEROSIS PATIENTS UTILIZING A NEW FUNCTIONAL
CELL BASED IFN-A REPORTER GENE ASSAY
Martins TB1, Rose JW3, Jaskowski T1, Kusukawa N1, Gardiner GL1,
Husebye D3, Seraj HS3, Hill HR1,2 1ARUP Institute for Clinical and
Experimental Pathology, SLC, UT; 2University of Utah School of Medicine,
SLC, UT and 3University of Utah School of Medicine, SLC, UT.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Purpose of Study: Interferon (IFN)-A is well established as first line
therapy in relapsing/remitting multiple sclerosis (MS). Patients with therapy
induced neutralizing antibodies (Nabs) to IFN-A have shown reduced
responses to treatment with IFN-A, having higher relapse rates, increased
MRI activity and a higher risk of disease progression.
Methods Used: We employed a new functional assay which measures
IFN-A neutralizing antibody utilizing a division arrested, cryopreserved
human cell line which have been transfected with a luciferase reporter gene.
Summary of Results: This new IFN-A reporter gene assay demonstrated
100% sensitivity compared with the traditional cytopathic effect assay
(CPE), correctly identifying 25 of 25 samples as positive for the presence
of IFN-A neutralizing antibody. Specificity was also 100% with 58 of 58
normal donors testing negative. Additionally, 165 multiple sclerosis patients
undergoing IFN-A therapy were tested for IFN-A neutralizing antibody by
the IFN-A reporter gene assay. The percent of patients testing positive for
neutralizing antibodies to IFN-A were, according to the IFN-A therapy
utilized: Avonex (1>) 34.3%, Rebif (1>) 44.7%, and Betaseron (1A) 38.1%.
The IFN-A cell surface receptor did show slight cross-reactivity in samples
spiked with high concentrations (1000 pg/ml) of recombinant IFN-F, but
showed no cross-reactivity to interleukin (IL)-1A, 2, 4, 5, 6, 8, 10, 12, 18,
and TNF->.
Conclusions: The IFN-A reporter gene assay showed excellent correlation
with the well established CPE assay and also offered clear advantages.
Results can be reported within 20 hours compared to 4Y6 days for the CPE
assay, the cell line has been well characterized and is cryopreserved assuring
better run to run reproducibility, and there is no need to maintain a viral
culture system. The use of this new functional assay should be a valuable tool
for the monitoring of IFN-A treated MS patients for the presence of neutralizing antibodies, hopefully leading to more effective therapy.
354
INVESTIGATION OF STAT3-DEPENDENT NEUTROPHIL
MIGRATION IN JOB SYNDROME
Kumánovics A1,3, Augustine NH1,3, Hill HR1,2,3 1Univ of Utah, SLC, UT;
2
Univ of Utah, SLC, UT and 3ARUP Institute for Clinical and Experimental
Pathology, SLC, UT.
Purpose of Study: Hyper-immunoglobulin E (IgE) syndrome (HIES), or
Job syndrome is a primary immunodeficiency, characterized by high serum
concentration of IgE, eczema, eosinophilia, indolent cold abscesses, candidiasis, and pneumonias complicated by pneumatocele formation and
bronchiectasis. In addition to the immune system defects, HIES is also
characterized by connective tissue, skeletal and dental development defects.
Mutations in STAT3 gene have been identified in classical autosomal dominant HIES patients. STAT3 participates in the signal transduction of many
cytokines and hormones, and thus provides a possible explanation for the
diverse dysfunctions. However, direct connections between the various
clinical manifestations of HIES and STAT3 mutations have not yet been
clearly established. Chemotactic defects in the peripheral blood neutrophils
from HIES patients was first described by us and verified by other investigators. Many noted that the chemotactic defect is a variable feature of the
disease, but these early studies used a variety of methods and chemotactic
agents to study neutrophil migration. The discovery of the genetic etiology
of autosomal dominant HIES makes it possible to re-examine neutrophil
chemotaxis in molecularly-defined HIES patients using chemokines affecting
STAT3-dependent and independent signaling pathways.
Methods Used: Chemotaxis of peripheral blood neutrophils from HIES
patients and healthy donors were examined in response to various stimuli
in multiwell chemotaxis chamber with 5-Km pore-size filters. Actin polymerization was detected by measuring FITC-phalloidin fluorescence intensity by flow cytometry.
Summary of Results: Neutrophils from HIES patients have a chemotaxis
and actin polymerization defect in response to CXCL1 and CXCL2, but
normally respond to the complement component C5a, whereas the response
to formylated peptide fMLP is variable. CXCL1 and CXCL2 signals are
mediated by the CXCR2 chemokine receptor. CXCR2, like the receptors
for fMLP and C5a, belongs to the G protein-coupled receptor family, but it
is unique in that it can reportedly activate the STAT3 signaling system.
Conclusions: Our results demonstrate that neutrophils from HIES patients
have an intrinsic chemotaxis defect in response to chemotactic signals mediated by CXCR2.
* 2011 The American Federation for Medical Research
355
EFFECTS OF A NON-STEROIDAL ANTI-INFLAMMATORY
DRUG ON SUPPRESSED ACTIVITY IN MICE SUFFERING
FROM CHRONIC JOINT INFLAMMATION
Suhail M1, Christianson C2, Koehrn F2, Malkmus S2, Corr M3, Yaksh TL2
1
UCSD, La Jolla, CA; 2UCSD, La Jolla, CA and 3UCSD, San Diego, CA.
Purpose of Study: Chronic pain has profound effects on activity and
sleep-wake cycles. In chronic cancer and arthritic patients, there is a decrease in activity secondary to pain. Previous reports indicate chronic inflammatory conditions result in reduced activity which normalizes upon
pain treatment. However, there has been little systematic investigation of
this process.
Methods Used: Rheumatoid arthritis (RA) is an autoimmune disorder
that causes significant joint pain. The K/BxN serum transfer model has
been characterized both as a model for RA and chronic inflammatory pain.
Transfer of serum from K/BxN mouse to naBve mice will initiate an autoimmune response against glucose-6-phosphate isomerase present in joints,
leading to the development of severe inflammatory arthritis. Here, we investigated the activity of mice following K/BxN serum transfer vs. control
serum and observed the activity changes following delivery of an NSAID
(ketorolac).
Experimental protocols were approved by the Institutional Animal Care
committee. Male C57Bl/6 mice were injected i.p. on days 0 and 2 with 100HL
of K/BxN serum or normal control serum. Activity was monitored by motion
sensors on individual cages and each mouse_s activity was corrected according to baseline pre-treatment values.
Summary of Results: Arthritic mice had significantly decreased activity
on nights 0Y11 (pG.05), after which they normalized. Arthritic K/BxN
serum transfer mice were treated with ketorolac PO (approximately 10mg/kg,
15mg/kg, or 20mg/kg) in their drinking water for nights 6Y8. All doses
resulted in increased activity compared to untreated K/BxN arthritic mice
(pG.05, pG.001, pG.001, respectively). After ketorolac was stopped, animals
receiving 15 and 20mg/kg maintained elevated activity levels as compared
to control though day 12 (pG0.05).
Conclusions: This suggests a long-term behavioral or disease-modifying
effect of ketorolac on inflammatory pain during K/BxN serum transfer arthritis. Furthermore, these results indicate the efficacy of diurnal activity
monitoring to better investigate behavior in uncontrolled pain states and
new therapies for pain.
356
SEVERE URTICARIA AND ABDOMINAL PAIN DUE
TO ANTIBODIES TO IGE RECEPTOR TREATED
WITH CYCLOSPORINE
Wang KY, Roberts RL UCLA, Los Angeles, CA.
Purpose of Study: Urticaria and abdominal pain due to antibodies to the
IgE Fc Receptor was successfully treated in children with oral cyclosporine.
Methods Used: We described 2 children with urticaria due to antibodies
to IgE Fc receptor. The first patient started to develop severe, painful urticaria lesions with abdominal pain at 5 years. Work-up showed no evidence
of allergies (IgE 58 IU/ml) but positive for antinuclear antibody (1:320) and
thyroid antibodies. The urticaria only responded to prednisone. She was
found to be positive for IgE receptor antibody (23.5%). The second patient
developed severe urticaria and severe abdominal pain at 14 years. She also
had a low IgE (38 IU/ml) but positive for ANA (1:160). The lesions only
responded to daily prednisone (25 mg/daily). She also had antibodies to IgE
receptor (64%).
Summary of Results: The first patient was started on cyclosporine (initially 150 mg/daily) and we were able to taper off completely the prednisone
within 2Y3 months. The urticarial lesions completely resolved as did the
abdominal pain.
The second patient was also started on cyclosporine (175 mg/day) and we
were able to taper her prednisone to zero without the recurrence of urticaria
in 3 months. Her abdominal pain also resolved.
Conclusions: Children with severe urticaria due to antibodies to IgE receptor went into complete remission with oral cyclosporine. Their abdominal
pain also resolved with cyclosporine, suggesting that the pain could have
also due to the antibodies to IgE receptor which may have caused local
abdominal pain by histamine release in the gut.
187
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Baseline pain scores and demographics were com357
A RANDOMIZED CONTROLLED TRIAL OF
COST-EFFECTIVENESS OF ULTRASOUND-GUIDED
INTRAARTICULAR INJECTION OF INFLAMMATORY
ARTHRITIS
Norton H1, Sibbitt WL1, Bankhurst A1, Delea S1, Chavez-Chiang N1, Band
P2 1University of New Mexico, Albuquerque, NM and 2New York University
School of Medicine, New York, NY.
Purpose of Study: The present randomized controlled study addressed
whether sonographic needle guidance affected the outcomes of intraarticular
injection for inflammatory arthritis.
Methods Used: 244 symptomatic joints (3% small, 51% intermediate, and
46% large joints) with inflammatory arthritis (76% rheumatoid arthritis)
were-randomized to injection by conventional palpation-guided anatomic
injection (120 joints) or sonographic image-guided injection enhanced with
a-one-handed RPD (the reciprocating procedure device) mechanical syringe
(124 joints). A one needle, two-syringe technique was used. After intraarticular placement and synovial space dilation were confirmed by sonography, a
syringe exchange was performed, and triamcinolone acetonide was injected
with the second syringe through the indwelling intraarticular needle. Baseline
pain, procedural pain, pain at outcome (2 weeks and 6 months), responders,
therapeutic duration, reinjection rates, total cost, and cost per responder were
determined.
Summary of Results: Relative to conventional palpation-guided methods,
sonographic guidance for injection of inflammatory arthritis resulted in 81%
reduction in injection pain (pG0.001), a 35% reduction in pain scores at outcome (pG0.02), 38% increase in the responder rate (pG0.003), 34% reduction
in the non-responder rate (pG0.003), a 32% increase in therapeutic duration
(p =0.01), a 8% reduction ($7) in cost/patient/year, and a 33% ($64) reduction
in cost/responder/year for a hospital outpatient (pG0.001).
Conclusions: Sonographic needle guidance improves the performance,
clinical outcomes, and cost-effectiveness of intraarticular injections of inflammatory arthritis.
358
TECHNIQUE-BASED IMPROVEMENTS IN THE
COST-EFFECTIVENESS OF INTRAARTICULAR INJECTIONS
FOR OSTEOARTHRITIS OF THE KNEE: A RANDOMIZED
CONTROLLED STUDY
Norton H1, Band P2, Sibbitt WL1, Chavez-Chiang N1, Delea S1, Bankhurst
A1 1University of New Mexico, Albuquerque, NM and 2New York University,
New Yord, NY.
Purpose of Study: This randomized controlled study compared the costeffectiveness of two different low-cost, palpation-directed intraarticular injection techniques for osteoarthritis (OA) of the knee.
Methods Used: 128 symptomatic osteoarthritic knees were randomized
to palpation-guided intraarticular injection with either 1) a conventional syringe (CS), or 2) a single-handed aspiration syringe (AS), the RPD (the
reciprocating procedure device). 3 ml of 1% lidocaine were used for anesthesia, followed by arthrocentesis, and then injection of 80 mg of triamcinolone acetonide. Baseline pain, procedural pain, aspirated fluid volume, pain
at outcome (2 weeks and 6 months), responder rates, time to reinjection, cost/
patient/year and cost/responder/year were determined.
188
parable between the groups (p90.2 for all). Both techniques significantly
reduced pain scores at outcome from baseline (p G0.001). Relative to the CS
technique, the AS technique resulted in a 110% greater volume of aspirated
fluid, a 37% reduction in procedural pain (pG0.01), a 43% greater reduction
in pain scores at outcome (pG0.01), a significant increase in responder rate
(pG0.01), a significant reduction in non-responder rate (pG0.01), 29.7% increase in the time to next injection (pG0.01), and a 23% reduction ($28 US)
in cost/patient/year (pG0.001).
Conclusions: Utilization of a low-cost, single-handed aspiration syringe
improved outcomes and reduced overall costs or intraarticular injections of
osteoarthritis of the knee. Larger multicenter studies are needed to confirm
these results in diverse practice settings.
359
NEW DEVICE TECHNOLOGIES FOR SUBCUTANEOUS
FAT BIOPSY
Kettwich LG1, Sibbitt WL1, Chavez-Chiang N1, Delea S1, Michael A2,
Bankhurst A1 1University of New Mexico Health Sciences Center,
Albuquerque, NM and 2Texas Tech Health Sciences Center, El Paso, TX.
Purpose of Study: Subcutaneous fat biopsy by needle aspiration is useful
for the evaluation of amyloidosis, environmental contaminants, lipid metabolism, genetic studies, and diabetes research; however, aspiration with conventional syringes is awkward and can result in needle trauma to patient tissues.
The present study examined new technologies for needle aspiration biopsy.
Methods Used: Subcutaneous fat biopsy in 10 high-risk individuals with
arthralgias and neuropathic symptoms was randomized to 1) a 10 ml RPD
(reciprocating procedure device) mechanical syringe, or 2) a 60 ml vacuum
syringe. In each case an 18 gauge 2 inch (5 cm) Chiba biopsy needle
was utilized. Outcome measures included patient pain by the 10 cm Visual
Analogue Pain Scale (VAS), adequacy of biopsied tissue, complications, and
diagnosis. The operator_s ability to control the syringe was quantitatively
measured by the linear displacement method.
Summary of Results: Both the vacuum and mechanical syringes permitted facile aspiration of subcutaneous fat; in all cases, there was adequate
sample obtained for analysis without complications. The mechanical and the
vacuum syringes enhanced control of the needle compared to conventional
syringes, reducing unintended forward penetration by 75% (3.6T0.5 mm)
and 87% (12.0T1.4 mm), respectively (pG0.0001). Although adipose cells
were obtained in abundance, small blood vessels and connective tissue septa
were also obtained intact permitting precise microhistological examination.
One case of primary AL amyloidosis (kappa light chain disease) was diagnosed in each group.
Conclusions: Subcutaneous fat biopsy by needle aspiration can be facilely
achieved with new aspiration syringe technologies with improved needle
control and enhanced patient safety.
360
MIDDLE CEREBRAL ARTERY RESISTIVITY AND
PULSATILITY INDICES IN SYSTEMIC LUPUS
ERYTHEMATOSUS
Boyer NM1, Roldan C1, Yonan KA2, Sharrar JM1, Sibbitt WL1, Greene ER1,2
1
University of New Mexico, Albuquerque, NM and 2New Mexico Highlands
University, Las Vegas, NM.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Purpose of Study: Systemic lupus erythematosus (SLE) is associated with
significant cerebrovascular and neuropsychiatric disease for which multiple
pathogenesis have been proposed. Although global and focal cerebral
hypoperfusion have been proposed, there are limited data about intracerebral
arterial hemodynamics. Transcranial Doppler (TCD) allows portable, high
temporal (6 ms) and spatial (0.3 mm) resolution, and noninvasive blood
velocity measurements in the middle cerebral arteries and calculations of
standard resistivity (RI) and pulsatility (PI) indices. Determined from the
blood velocity waveforms, RI and PI correlate linearly with cerebral hemispheric arteriolar tone, blood flow resistances, and impedances. Accordingly,
we hypothesized that there would be a significant differences (pG0.05) in RI
and PI between SLE patients and healthy, age and gender matched controls.
With similar blood pressures, decreased RI and PI would be consistent with
a compensatory mechanism to maintain relative hyper or normal cerebral
perfusion.
Methods Used: Thirty four stable SLE patients (35T11 years) and 15
control subjects (34T10 years) underwent TCD to measure RI and PI bilaterally. Patients and controls had simultaneously measured similar, normal
blood pressures and were examined in the supine position during normal,
resting respiration (minimize PCO2 changes). RI and PI were determined by
a single observer blinded to all subjects_ clinical data.
Summary of Results: SLE patients had lower (pG0.05) average RI values
as compared to controls: 0.45T0.10 versus 0.52T0.05; respectively. Similarly,
SLE patients had lower (pG0.05) average PI than controls: 0.65T0.19 versus
0.77T0.12; respectively. There were no significant differences in RI and PI
bilaterally within each cohort.
Conclusions: Easily measured and robust RI and PI values in the middle
cerebral artery are significantly lower in SLE as compared to controls. These
indirect indices suggest that middle cerebral arterial resistances and impedances are decreased in SLE, and under normotensive conditions, are consistent with increased arteriolar dilation, increased medium size arterial
cerebral blood flow, and a compensatory mechanism for hyper or normal
cerebral perfusion in SLE.
Metabolism II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
361
ISLET MATRIX METALLOPROTEINASE-9
IS DOWNREGULATED IN A MOUSE MODEL OF
TYPE 2 DIABETES
Svy D1,2, Aston-Mourney K2,1, Kahn S2,1 1University of Washington,
Seattle, WA and 2VAPSHCS, Seattle, WA.
Purpose of Study: A pathological hallmark of the pancreatic islet in type
2 diabetes is aggregation of islet amyloid polypeptide (IAPP) into extracellular amyloid deposits, the process of which is toxic to A cells. We have
previously shown that (a) glucose dose-dependently increases amyloid deposition in human IAPP (hIAPP) transgenic (T) mouse islets, (b) matrix
metalloproteinase (MMP)-9 degrades hIAPP, and (c) inhibition of MMP-9
activity increases amyloid deposition in hIAPP T islets. Whether high glucose regulates MMP-9 or tissue inhibitor of metalloproteinase (TIMP)-1, a
natural inhibitor of MMP-9, is unknown. Thus, we determined if downregulation of MMP-9 occurs in islets in vitro when cultured in high glucose
and/or in vivo in a diabetic mouse model.
Methods Used: Amyloid-forming hIAPP T and non-amyloid forming nontransgenic (NT) mouse islets were cultured for 7 days in 11.1, 16.7, or 33.3 mM
glucose (n=4Y6). Islets were also isolated from diabetic db/db and control db/+
mice (n=8). To confirm the presence of amyloid in T islets after culture, islet
sections were stained with thioflavin S. MMP-9 and TIMP-1 mRNA levels
were determined by real-time PCR using 18S as the endogenous control.
MMP-9 activity was measured in islet conditioned medium by gelatin zymography (n=3).
Summary of Results: In cultured T and NT islets, neither MMP-9 nor
TIMP-1 mRNA changed with increasing glucose. In contrast, MMP-9 activity tended to increase at 33.3 mM glucose (NT at 11.1, 16.7, and 33.3 mM
glucose: 1, 0.98T0.21, 3.61T0.95; T at 11.1, 16.7, and 33.3 mM glucose:
1.22T0.06, 1.03T0.18, 3.92T1.60; p=0.06), but was not affected by amyloid
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
formation which was observed at 16.7 and 33.3 mM glucose. In db/db mouse
islets, MMP-9 mRNA decreased by 41% when compared to db/+ islets
(p=0.04).
Conclusions: Glucose did not downregulate MMP-9 activity in cultured
islets and therefore, decreased MMP-9 is not the mechanism by which glucose induces amyloid formation in this model. However, in a whole animal
model, MMP-9 mRNA was significantly decreased in diabetes. This suggests
that downregulation of MMP-9 by another component of the diabetic milieu,
such as elevated free fatty acids, may contribute to the increased amyloid
deposition associated with type 2 diabetes.
362
INCREASED EXPRESSION OF THE VASOCONSTRICTOR
ENDOTHELIN-1 MAY CONTRIBUTE TO ISLET
ENDOTHELIAL DYSFUNCTION IN DIABETES
Ottley A1, Kahn S2,1, Hull R2,1 1University of Washington, Seattle, WA and
2
VAPSHCS, Seattle, WA.
Purpose of Study: Increased production of the vasoconstrictor endothelin-1
(ET-1) and decreased activity of the vasodilation-promoting enzyme prostacyclin synthase (PGIS) contribute to diabetic macrovascular endothelial dysfunction. We have demonstrated that the pancreatic islet is also affected by
diabetic endothelial dysfunction. We thus determined if ET-1 and PGIS are
expressed in islets and whether islet ET-1 expression increases and PGIS expression decreases with diabetes.
Methods Used: ET-1 and PGIS mRNA levels were measured by real-time
PCR in immortalized murine endothelial cells (MS-1) and A cells (A-TC3),
and whole islets. Islets were isolated from diabetic C57BL/6.db/db mice or
non-diabetic db/+ mice at 8 and 16 weeks of age. E-selectin was used as a
marker of endothelial dysfunction, PECAM as a marker of endothelial cell
number, and insulin as a marker of A-cell number.
Summary of Results: ET-1 mRNA was detectable in MS-1 cells. PGIS
mRNA was detectable in both MS-1 and A-TC3 cells. As expected, E-selectin
and PECAM were strongly expressed in MS-1 cells and insulin was strongly
expressed in A-TC3 cells. At the time of islet isolation db/db mice were hyperglycemic compared to db/+ controls (Table). ET-1 mRNA levels tended to
increase in db/db islets at 16 weeks (Table; p=0.09), while PGIS mRNA
levels did not significantly differ from control at 8 or 16 weeks. As expected,
E-selectin mRNA levels were increased in db/db islets at 16 weeks, while
PECAM mRNA levels were decreased at both 8 and 16 weeks.
Conclusions: ET-1 is selectively expressed in islet endothelial cells while
PGIS is expressed in both islet endothelial and A cells. If PGIS activity
decreases in diabetes it is not via decreased mRNA levels. Increased islet
expression of the vasoconstrictor ET-1 occurs with prolonged diabetes and
may, therefore, contribute to islet endothelial dysfunction. This, in turn, may
contribute to islet dysfunction in type 2 diabetes.
Data are meanTSEM. *pG0.05, **pG0.01 vs db/þ age-matched control.
363
OXIDATIVE STRESS PROMOTES ISLET
AMYLOID DEPOSITION
Morcos M1,2, Zraika S2,1, Kahn S2,1 1University of Washington, Seattle, WA
and 2VAPSHCS, Seattle, WA.
Purpose of Study: Type 2 diabetes and chronic hyperglycemia are associated with oxidative stress in pancreatic A cells. Islet amyloid, a pathological
feature of the pancreas in type 2 diabetes, contributes to the loss of A-cell
mass but its relationship to oxidative stress is not fully understood. Previous
studies have shown that amyloid deposition increases oxidative stress in
189
Western Regional Meeting Abstracts
Journal of Investigative Medicine
islets; however, extended exposure to amyloid-induced oxidative stress may
also increase amyloid deposition in a positive feedback manner. Therefore, oxidative stress may promote amyloid deposition and the loss of
A-cell mass.
Methods Used: We used an in vitro model of islet amyloid formation where
human islet amyloid polypeptide (hIAPP) transgenic mouse islets produce
amyloid in a glucose-dependent manner and non-transgenic islets are incapable of producing amyloid. Islets were incubated in 11.1 or 16.7 mmol/L
glucose for 7 days with or without 0.5 mM glyceraldehyde, a known oxidative
stressor. Oxidative stress, amyloid and A-cell area were visualized using nitrotyrosine, thioflavin S and insulin staining respectively. Amyloid severity was
calculated as the proportion of islet area occupied by amyloid and A-cell area
was calculated as the proportion of islet area occupied by insulin.
Summary of Results: Glyceraldehyde treatment increased oxidative stress
in both 11.1 and 16.7 mmol/L glucose-cultured islets. In the 16.7 mmol/L
glucose-cultured hIAPP transgenic islets, treatment with glyceraldehyde did
not significantly increase amyloid deposition (2.91 T 0.84% vs. 2.65 T 0.45%;
p=0.66, n=5), nor did it change A-cell area (66.1 T 2.0 % vs. 67.2 T 2.4%;
p=0.21, n=5). These findings suggest that in islets with amyloid-induced
oxidative stress, further induction of oxidative stress with glyceraldehyde
does not promote additional amyloid formation or A-cell loss. Conversely, in
the 11.1 mmol/L glucose-cultured hIAPP transgenic islets, where amyloid
deposition is normally minimal, glyceraldehyde treatment increased amyloid
deposition (0.29 T 0.10 % vs. 1.22 T 0.33%; p=0.02, n=5) and reduced A-cell
area (71.1 T 0.8% vs. 68.6 T 0.7%; p=0.01, n=5). As expected, the nontransgenic islets did not form amyloid.
Conclusions: The induction of oxidative stress under conditions of minimal
islet amyloid deposition promotes amyloid formation and may contribute to
the loss of A-cell mass in type 2 diabetes.
364
NO MEDIATED REGULATION OF CREB EXPRESSION
Nelson SK, Miller M, Knaub L, Gunter J, Reusch JE University of
Colorado, Denver, CO.
Purpose of Study: CREB (cAMP response-element binding protein) is a
transcription factor that maintains vascular smooth muscle cell (VSMC)
quiescence which is down-regulated in a variety of states in vivo (including
diabetes mellitus, pulmonary hypertension, dyslipidemia and metabolic
syndrome) and in vitro (with oxidative stress, glucose, oxLDL, and PDGF).
Degradation of CREB in vivo and in vitro is accompanied by a decrease in
anti-oxidant capacity, mitochondrial expression and survival. Hypothesis:
Oxidative stress is responsible for increasing CREB expression in acute
settings for maintenance of cellular homeostasis, but also responsible for
decreases in CREB seen chronically. Previously, our lab has shown that LNAME, a NOS (nitric oxide synthase) inhibitor, decreased CREB expression
and increased CREB ubiquitination in vivo, leading us to propose that induction of CREB in response to oxidative stress may be regulated via NOS
production of NO.
Methods Used: Primary VSMC cultures; endothelial NOS (eNOS)
KO mice.
Summary of Results: Primary VSMC cultures were treated with H2O2 to
generate oxidant stress. Over 4 hours following treatment, CREB was initially activated, significantly depleted and then recovered. Neuronal NOS
(nNOS) protein expression increases significantly in parallel with CREB at
the 4 hour recovery time point. Treatment of VSMCs with L-NAME produced similar significant biphasic changes. Preliminary studies in VSMCs +/oxidative stress +/- SNAP (an NO donor) suggest that SNAP can rescue
expression of CREB acutely. Interestingly, aortic lysates from endothelial
NOS (eNOS) KO mice, which we hypothesized would have decreased
CREB expression due to a lack of eNOS generated NO in the endothelium,
showed increased CREB and CREB-target expression compared to agematched wild-types at 8 weeks, while at 18 weeks, CREB activation was
increased but overall CREB content decreased, suggesting that another
pathway might be compensating. Summary: Oxidative stress produces significant changes in CREB expression over a 4-hour time period, nNOS expression parallels the expression pattern of CREB and inhibition of NOS
with L-NAME produces the same effect. Genetic deletion of eNOS in mice
produces an increase in CREB at 8 weeks but a decrease at 16 weeks.
Conclusions: NOS regulated NO production modulates CREB content and
function in response to oxidative stress.
190
&
Volume 59, Number 1, January 2011
365
NICOTINE WORSENS HIGH FAT DIET- INDUCED HEPATIC
AND MUSCLE STEATOSIS IN MICE
Nzenwa I1,2, Vides R1,3, Mangubat M1, Garcia L1, Shen R1, Sinha-Hikim
AP1,2, Friedman TC1,2, Sinha-Hikim I1,2 1Charles Drew University, Los
Angeles, CA; 2David Geffen School of Medicine-UCLA, Los Angeles, CA
and 3Stanford University, Stanford, CA.
Purpose of Study: Smoking is a major risk factor for many diseases.
There is evidence that smoking may contribute to non alcoholic fatty liver
disease (NAFLD) and skeletal muscle abnormalities. The health risk associated with smoking is exaggerated by obesity and is the leading causes of
morbidity and mortality worldwide. We hypothesize that nicotine can have
additive effects on HFD-induced hepatic steatosis and skeletal muscle abnormalities in obese mice.
Methods Used: Adult C57BL6 male mice were fed with normal diet (ND)
or HFD with 60% of calories derived from fat (Research Diets, New
Brunswick, NJ) with or without twice daily injections of 0.75 mg/kg BW of
nicotine for 10 weeks.
Summary of Results: Compared with mice on ND, mice fed with HFD
exhibited significant weight gain and increased abdominal as well as epididymal fat mass. Nicotine-treated mice on a HFD showed significantly less
weight gain than mice fed a HFD alone. Abdominal fat and overall fat content
was substantially reduced as determined by DEXA scan. Food intake was
unchanged. Nicotine-treated mice on a HFD also had greater oxidative stress,
as indicated by low GSH/GSSG ratio in comparison to HFD alone. Light and
electron microscopy revealed increased lipid accumulation of varying sizes
in hepatocytes from mice on a HFD. Addition of nicotine to HFD resulted in
a further increase in lipid accumulation and in the size of lipid droplets.
Intriguingly, compared with mice fed with HFD alone, where no or minimal
lipid droplets were detected, combined treatments of nicotine and HFD resulted in intramitochondrial fat (IMF) accumulation in myofibers. We further
observed mitochondrial swelling and vacuolization after combined treatment
with nicotine and HFD than of the mice on HFD alone.
Conclusions: These results indicate that nicotine further accelerates hepatic
steatosis and skeletal muscle abnormalities induced by HFD and provide a basis
for further studies aimed to elucidate the mechanisms of additive effects of
nicotine and HFD on NAFLD and IMF accumulation in skeletal muscle. We
surmise that nicotine plus HFD is likely to be a very toxic combination in
patients.
366
MONOUNSATURATED FATTY ACIDS INCOMPLETELY
RESCUE 3T3-L1 CELLS FROM EFFECTS OF
SCD1 INHIBITION
Yee JK, Wahjudi P, Patterson ME, Lim S, Mao C, Lee W Los Angeles
BioMedical Research Institute at Harbor-UCLA, Torrance, CA.
Purpose of Study: Enhanced adipogenesis during adipocyte differentiation contributes to obesity development. Stearoyl-CoA desaturase enzyme 1
(SCD1) converts the saturated fatty acids palmitate and stearate to the
monounsaturated fatty acids (MUFAs) palmitoleate and oleate, respectively.
SCD1 is induced during adipocyte differentiation, is upregulated in obesity,
and is a potential target for obesity prevention. Oleate can induce lipid accumulation before differentiation gene induction. The objectives are: 1) to
determine the effects of SCD1 inhibition during adipocyte differentiation on
desaturation and cell proliferation using stable isotopes; 2) to determine if
the effects of SCD1 inhibition are altered by addition of MUFA.
Methods Used: Confluent 3T3-L1 preadipocytes in culture were treated
with or without differentiation induction cocktail in the medium for four
days. Differentiating cells were further treated in 4 groups during induction:
1) no additional treatment 2) SCD1 inhibitor CGX0290 3) CGX0290 and
palmitoleate or 4) CGX0290 and oleate. In the first two days of induction,
all groups received medium with 25% [U13C]glucose. After induction was
completed, they received medium with [U13C]stearate. Cells were harvested
and counted on day 8, fatty acid and DNA isotopomers were analyzed by GC/
MS. The 13C-enriched oleate/stearate ratio was calculated as the desaturation
index (DI), a measure of SCD1 activity.
Summary of Results: CGX0290 decreased the DI based on isotopomers
of newly formed fatty acids. Addition of palmitoleate to CGX0290 parially
restored the DI, but addition of oleate decreased the DI further. After induction
was completed, the DI as measured by desaturation of [U13C]stearate was
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
increased in the CGX0290 group. Cell counts were decreased in the CGX0290
group, although the 13C DNA enrichment was increased.
Conclusions: CGX0290 is an effective SCD1 inhibitor. CGX0290 does
not prevent differentiation of 3T3-L1 cells, but decreases cell numbers likely
through increased apoptosis. Addition of MUFA does not reverse effects of
SCD1 inhibition, and oleate appears to feedback inhibit SCD1 further. Effects
of SCD1 inhibition may be sensitive to modulation by nutrition. Potential use
of CGX0290 in obesity prevention or treatment definitely requires further
investigation.
367
IS 1,5 ANHYDROGLUCITOL (GLYCOMARK\)
A CLINICALLY USEFUL TEST IN TYPE 1 DIABETES?
Duran-Valdez E1, Burge M1, Broderick P2, Shey L1, Valentine V3, Schade DS1
1
University of New Mexico, Albuquerque, NM; 2Presbyterian Hospital,
Albuquerque, NM and 3Diabetes Network, Albuquerque, NM.
Purpose of Study: Glycomark\ is an FDA approved test that measures 1,5
anhydroglucitol concentration in plasma. It is promoted as being a good
assessment of postprandial hyperglycemia in diabetes. However, its clinical
utility in type 1 diabetes has not been established. We utilized established
glucose parameters, both in the blood and the interstitial fluid, to assess its
ability to predict glucose control in type 1 diabetes.
Methods Used: Eighty-four studies were performed in type 1 diabetic
volunteers. A fasting blood sample was drawn for both A1C and the 1,5
anhydroglucitol assay. We then performed five days of blinded-Continuous
Glucose Monitoring (CGM) with simultaneous capillary blood glucose
measurements. The data supplied from the CGM recording device was correlated with the 1,5 anhydroglucitol value. The following parameters were
examined: blood A1C, mean capillary blood glucose, mean interstitial glucose above 180mg/dl, mean postprandial integrated interstitial glucose, percent time spent above 180mg/dl, and mean interstitial glucose.
Summary of Results: The first four of the parameters are shown below,
correlated with the 1,5 anhydroglucitol value. None of the correlations was
strong enough to be clinically useful.
Conclusions: 1,5 anhydroglucitol (Glycomark\) did not demonstrate any
significant correlation to commonly accepted glucose control parameters. 1,5
anhydroglucitol should not be used to draw conclusions about other systemic
glucose parameters until its reliability and clinical utility have been established in patients with diabetes.
368
A NOVEL MECHANISM TO PREVENT CARDIAC
CONSEQUENCES OF IN UTERO COCAINE EXPOSURE
Vides R1,3, Nzenwa I1,2, Shen R1, Sinha Hikim AP1,2, Sinha-Hikim I1,2
1
Charles Drew University, Los Angeles, CA; 2David Geffen School Of
Medicine-UCLA, Los Angeles, CA and 3Stanford, Stanford, CA.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Purpose of Study: Abuse of cocaine during pregnancy exposes ~ 100,000
infants per year to cocaine in the United States. Cocaine readily crosses
the placenta into the fetal circulation affecting the placenta and fetus with
numerous adverse outcomes, including fetal cardiac myocyte death. This
study was designed to test the hypothesis that minocycline, a second generation tetracycline, prevents fetal cardiac myocyte death induced by prenatal
cocaine exposure through inhibition of c-Jun NH2-terminal kinase (JNK)
and p38 mitogen-activated protein kinase (MAPK)-mediated mitochondria
intrinsic pathway signaling.
Methods Used: Timed mated pregnant SD rats received one of the following treatments twice daily from gestational day 15 to 21: (i) IP injections
of saline (control); (ii) IP injections of cocaine (15 mg/kg BW); and IP
injections of cocaine + oral administration of 25 mg/kg BW of minocycline.
Neonatal pups were killed on day 15 after birth. Additional pregnant dams
received twice daily IP injections of cocaine (from days 15 to 21 of gestation) + oral administration of 37.5 mg/kg BW of minocycline. Minocycline
treatment continued from day 15 of gestation until the pups were killed on day
15 after birth. Pups were kept with their mothers and allowed to be breast-fed ad
libitum. Moms were not given cocaine after delivery.
Summary of Results: In utero cocaine exposure resulted in an increase
in oxidative stress and fetal cardiac myocyte apoptosis through activation
JNK and p38 MAPK-mediated mitochondria-dependent apoptotic pathway.
Continued minocycline treatment from day 15 of gestation until the pups
were killed significantly prevented oxidative stress, kinase activation, perturbation of BAX/BCL-2 ratio, caspase activation, and significantly attenuated
fetal cardiac myocyte apoptosis after prenatal cocaine exposure.
Conclusions: Our results show in vivo cardioprotective effects of minocycline in preventing fetal cardiac myocyte death after prenatal cocaine exposure. Minocycline with its proven clinical safety and its ability to cross
placental barrier and enter into the fetal circulation may become an effective
therapy for preventing cardiac consequences of in utero cocaine exposure.
369
DISCORDANCE IN LIPID ACCUMULATION AND LESION
SIZE: DETECTION WITH COMBINED OIL RED O AND
MOVAT_S PENTACHROME STAINING
Kim J1, Haw A1, Reifsnyder P2, Leiter E2, O’Brien KD1 1University of
Washington, Seattle, WA and 2The Jackson Laboratory, Bar Harbor, ME.
Purpose of Study: Atherosclerosis is quantified in murine models either
with a neutral lipid stain, such as Oil red O (ORO), or with a stain to identify
total lesion area, such as Movat_s pentachrome. In humans, lesion composition, especially lipids and macrophage contents, correlate more strongly
with clinical event risk than does lesion size. As plaque neutral lipid is both
extracellular and macrophage foam cell-associated, ORO identifies both
plaque lipid and foam cells.
Methods Used: To facilitate quantification of both the lipid/inflammatory
component and total lesion area on the same tissue section, we developed a
technique in which formal-sucrose-fixed, frozen tissue sections of murine
aortic sinuses were stained with ORO. Following photomicrography, ORO
was removed by rinsing in de-ionized H2O. Sections then were stained with
Movat_s pentachrome.
Summary of Results: The method then was applied to an experiment
assessing the effect of insulin-deficient diabetes due to the Ins2-Akita mutation on atherosclerosis in LDL receptor-deficient, C57BL/6J (B6.Ldlr)
mice. At 6 weeks of age, Ins2-Akita and WT B6.Ldlr mice (N=9/group)
were fed the atherogenic BPaigen[ diet (15% fat, 1.25% cholesterol, and
0.5% sodium cholate), for 14 weeks prior to sacrifice at age 20 weeks.
Average aortic sinus ORO and lesion areas were determined. At sacrifice,
plasma HDL levels were higher in Ins2-Akita vs. WT mice (P=0.044), but
levels of cholesterol, triglycerides, blood urea nitrogen or non-esterified fatty
acids did not differ between groups. Interestingly, lesion ORO area was
significantly lower in Ins2-Akita vs. WT mice (23.6T1.6 vs. 29.6T1.1 %,
P=0.0054), but total lesion areas did not differ (6.6T0.4 vs. 6.4T0.5 10-5
Km2, P=0.78). In addition, ORO and lesion areas were not correlated in
Ins2-Akita mice (R2=0.05, P=0.52) but were correlated strongly in WT
mice (R2=0.79, P=0.0032).
Conclusions: Thus, combined ORO/Movat_s staining efficiently quantifies
both plaque lipid and plaque lesion area. Use of this method in the Ins2-Akita
model demonstrates divergent effects on plaque lipid and plaque size, and
emphasizes the importance of assessing plaque composition in experimental
atherosclerosis.
191
Western Regional Meeting Abstracts
Journal of Investigative Medicine
370
IUGR ALTERS HEPATIC mRNA LEVELS OF HMGA1, RBP4,
AND INSR IN A GENDER SPECIFIC MANNER IN D120 RATS
Burshears B, Hale MA, Fu Q, Yu X, Callaway C, McKnight R, Joss-Moore L,
Lane RH University of Utah, Salt Lake City, UT.
Purpose of Study: IUGR individuals are at higher risk for adult-onset
metabolic disturbances, such as insulin resistance (IR). Furthermore, IUGR
induced by uteroplacental insufficiency in rats has been previously characterized by increased adiposity and IR, with males being more severely affected than females. High mobility group A1 (HMGA1), an architectural
transcription factor, contributes to or regulates the development of IR.
HMGA1 alters transcription of the insulin receptor gene (INSR) and retinol
binding protein 4 (RBP4), genes involved with IR. Hepatic expression of
HMGA1 and INSR is local, while liver secretes RBP4. It is unknown
what affect IUGR has on expression of HMGA1, INSR, and RBP4 in rat
liver. We therefore hypothesized that IUGR would decrease the expression
of HMGA1 and INSR in the rat liver. Additionally, we hypothesized that
RBP4 would be increased in IUGR.
Methods Used: IUGR was generated by bilateral uterine artery ligation on pregnant e19 rats. Using tissue from rat pup livers collected after
c-section at term (e21.5), or Day 0 (D0), D21, and D120. mRNA was isolated. cDNA was synthesized. RT-PCR was used to analyze HMGA1,
INSR, and RBP4.
Summary of Results: IUGR did not alter mRNA levels of HMGA1, INSR,
and RBP4 at D7 when compared to control. However, At D21, IUGR significantly decreased levels of RBP4 mRNA in males (pG0.05) but not in
females. IUGR increased RBP4 mRNA levels in D120 liver in males, but
not females. Moreover, female D120 mRNA levels of HMGA1 and INSR
in liver were increased when compared to control (pG0.05).
Conclusions: IUGR increased mRNA levels of HMGA1, INSR, and
RBP4 at D120 in a gender specific manner. Recent studies have shown
that increased adiposity correlates positively with RBP4 levels in males,
possibly through increased hepatic production. Therefore, we speculate
that decreased RBP4 levels in D21 males and increased RBP4 levels in
D120 males may represent the change in adiposity seen in our IUGR
model over time.
&
Volume 59, Number 1, January 2011
372
MEASUREMENT OF MOLECULAR DETERMINANTS OF
BODY WEIGHT REGULATION IN THE HUMAN BRAIN
USING MAGNETIC RESONANCE SPECTROSCOPY
Foster-Schubert KE1, Shankland E2, Frayo R1, Kushmerick M2, Conley K2,
Cummings D2 1University of Washington, Seattle, WA and 2University of
Washington, Seattle, WA.
Purpose of Study: Body weight is regulated such that alterations in bodyfat stores trigger compensatory changes in appetite and energy expenditure
that resist weight change. Increasing evidence from animal models demonstrates that hormonal and nutrient-related signals interact in key brain areas,
such as the hypothalamus, to regulate energy balance. For example, under the
influence of insulin and leptin, increases in intracellular long-chain fatty acylCoA (FACoA) lipids in hypothalamic neurons are hypothesized to convey a
state of energy abundance, thereby decreasing food intake. The relevance of
such pathways in the human brain is unknown because these molecules have
not been measured. The goal of our project is to measure FACoA in the human
brain using proton magnetic resonance spectroscopy (MRS).
Methods Used: 13 lean and obese healthy men and women will be studied
at baseline and following an intravenous injection of glucose and insulin to
stimulate FACoA. Serial hypothalamic MRS acquisitions will be obtained
over 1h to capture the FACoA peak. Fasting blood samples for glucose, free
fatty acids, insulin, leptin, and adiponectin will be obtained, as well as insulin,
glucose, and free fatty acids during the 1-h post-infusion period to model the
acute insulin response to glucose. We will determine whether changes in
peripheral biomarkers correlate with FACoA concentrations, and whether
these differ between lean and obese.
Summary of Results: Preliminary studies were completed to optimize
spectral acquisition parameters in human subjects. Using water suppression
(11139 mm3 voxel) with 4.8 min spectral acquisition binning, we detected metabolites at the concentration we hypothesize will be similar for
stimulated FACoA (Table).
Conclusions: Preliminary analyses indicate no competing lipid peak exists
in the region of interest for FACoA, and the voxel size and location does not
limit our ability to measure metabolites of comparable concentration for
stimulated FACoA.
371
A BIOASSAY OF INSULIN BIOACTIVITY USING HEPG2
CELLS IN CULTURE
Patterson ME1,2, Wahjudi P2, Yee JK1,2, Lee PW1,2 1Harbor-UCLA Medical
Center, Torrance, CA and 2LA BioMed, Torrance, CA.
Purpose of Study: Insulin is known to increase glucose phosphorylation
and glycogen synthesis in hepatocytes. The purpose of this experiment was
to develop a bioassay to examine the dose dependent effects of insulin on
the metabolism of glucose using HepG2 cells.
Methods Used: HepG2 cells were grown to a density of 25 10$6 cells/
mL, and were incubated in medium containing 100 mg/dL glucose, 10 mM
50% enriched U-13C3 lactate, and 1 mM sodium pyruvate for 4 hours. Three
sets of conditions were tested: A) no insulin, B) low dose insulin, and C)
high dose insulin. Medium glucose and cell glycogen quantitation were
carried out by GC/MS analysis with U-13C6 glucose as the internal standard
for the calculation of glucose uptake and glycogen synthesis. Contribution
from gluconeogenesis was assessed by determining 13C isotopomer in glycogen and medium glucose.
Summary of Results: Insulin stimulated glucose uptake in a dosagedependent manner. By contrast, glucose release as determined by the increase
in 13C label in medium glucose was decreased with increasing insulin concentrations. Glycogen production was increased with higher insulin concentrations. However, the fraction of labeled glucose in glycogen did not change
suggesting the relative contribution from glucose uptake and gluconeogenesis
to glycogen were not affected by insulin. Increasing insulin concentrations
also resulted in increased synthesis of fatty acids.
Conclusions: Consistent with the known metabolic effects of insulin on
glucose metabolism, we have shown that increasing concentrations of insulin are associated with increased glucose uptake, increased glycogen production, and decreased glucose release in HepG2 cells. The new assay system
gives additional insight into the role of insulin in the regulation of glucose
release and gluconeogenesis.
192
Neonatology Y Developmental Biology
Concurrent Session
8:00 AM
Saturday, January 29, 2011
373
MESENCHYMAL-SPECIFIC DELETION OF APC DISRUPTS
PARABRONCHIAL AND PERIVASCULAR SMOOTH
MUSCLE CELLS DIFFERENTIATION
Chan B, Li A, Li C, Xing Y, Li M, Minoo P Keck School of Medicine,
University of Southern California, Los Angeles, CA.
Purpose of Study: During embryonic development, Mesenchymal cells
that are programmed to become smooth muscle cell (SMC) elongate and
express alpha-smooth muscle actin (alpha-SMA). Dysregulation of SMC
proliferation and differentiation leads to debilitating pulmonary diseases including pulmonary hypertension and asthma. The Wnt signaling pathway
plays a key role in cell proliferation and cell fate determination during development. In the absence of a Wnt signal, Adenomatous Polyosis Coli
(APC) forms a destruction complex and inhibits beta-catenin activity. APC
also regulates cytoskeleton formation and controls cell migration. The role of
APC on SMC development is unknown. The current study was designed to
address this knowledge gap by examining the consequences of inactivating
the Apc gene and stabilizing beta-catenin via a Cre-LoxP approach, specifically in mesenchymal cells.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Methods Used: We used the Dermo1-cre driver line to induce recombination in Apc flox/flox mice. Double-transgenic Dermo1-cre; Apc flox/flox
embryos were sacrificed at different stages of development. The lungs were
analyzed by real-time PCR, western blot and immunohistochemistry.
Summary of Results: APC is expressed in normal developing lungs. APC
is co-localized with markers for smooth muscle and ciliated cells. APC deletion in mesenchymal cells leads to embryonic lethality at E12.5. In mutant
mesenchymal cells, immunoflourescent staining and western blot confirmed
a significant decrease in APC and a commensurate accumulation of betacatenin. The APC mutant lungs showed decreased parabronchial SMC differentiation. Gaps in the layer of alpha-SMA expressing cells surrounding
blood vessels were also found in the mutant embryos. Conditional stabilization of beta-catenin also generates a similar phenotype indicating that
the effect of Apc deletion on SMC differentiation is mediated via the Wnt
pathway.
Conclusions: APC plays a vital role in mesenchymal cell lineages during
late embryogenesis. Mesenchymal-specific deletion of APC blocks parabronchial and perviascular SMC differentiation. The effect of APC on the
SMC development is mediated via the beta-catenin-dependent Wnt signaling.
374
EFFECTS OF ERYTHROPOIETIN AND DARBEPOETIN ON
GROWTH AND DIFFERENTIATION OF CULTURED HUMAN
FETAL CARDIAC MYOCYTES
Stewart EM, McConaghy S, Ohls R University of New Mexico,
Albuquerque, NM.
Purpose of Study: Erythropoietin (Epo) is a regulator of differentiation of
erythroid progenitor cells. In myocardial tissue, Epo, through its interaction
with its receptor, reduces tissue ischemia by inhibiting apoptosis, dampening
of inflammatory response, and stimulation of angiogenesis. We evaluated the
effects of Epo and darbepoetin (Darbe), a long acting erythropoiesis stimulating agent (ESA), on cell growth, gene expression, and histological characterization of fetal cardiac myocytes between 10 and 18 weeks gestation. We
hypothesized that Epo receptor (EpoR) expression would increase with
gestational age, cell counts would increase with increasing ESA concentrations, and phenotype would not be altered by ESA exposure.
Methods Used: Primary myocyte cultures were grown following hand
suspension of 10Y18 week gestational age (GA) human fetal ventricular
cardiac tissue. Cells were incubated at 106cells/ml for 5 to 7 days in culture
medium containing 0, 10, or 100 units/ml Epo or in 1, 10, or 1000 ng/ml
Darbe. Cells were identified histologically and cell counts performed.
Myocyte phenotype was confirmed by positive troponin staining using
fluorescent microscopy, and myocyte progenitors were determined using
CD133, a glycoprotein expressed in cardiac progenitor cells. Total RNA
was isolated and quantified. RNA was reverse transcribed and quantitative
PCR performed using primers and probes to identify EpoR gene expression.
GAPDH was used as an internal control in duplex reactions.
Summary of Results: All cells stained positive for troponin, demonstrating
a myocyte phenotype. An inverse relationship was observed between the
number of cells with CD133 staining and increasing gestational age. There
was no significant difference in cell growth between cells grown in maximum
concentrations of Epo or Darbe compared to controls, and there was no
change in phenotype with ESA exposure. Epo receptor expression increased
with increasing ESA concentrations.
Conclusions: The percentage of progenitor cells in primary cultures of
human fetal myocytes decreased significantly with increased GA. Cells in
culture increased Epo receptor expression with increasing concentrations of
Epo or Darbe. Unlike other fetal tissue types, ESAs do not appear to be
mitogenic to fetal myocytes.
375
IUGR ALTERS MRNA LEVELS OF CHOLINERGIC
AND GABAERGIC RECEPTORS IN A
DEVELOPMENTALLY-SPECIFIC MANNER
Kunkel MD1, Wang Y2, Callaway C2, Yu X2, McKnight R2, Joss-Moore L2,
Lane RH2 1University of Utah, Salt Lake City, UT and 2University of Utah,
Salt Lake City, UT.
Purpose of Study: Intrauterine growth restriction (IUGR) causes abnormal
lung development leading to increased risk of impaired lung function in
adulthood. This is true in both humans and rats. In rat lungs, IUGR increases
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
mesenchymal thickness, which is associated with decreased apoptosis. Apoptosis, morphology, and cell proliferation are affected by cholinergic and
GABAergic autocrine and paracrine signaling systems. Nicotinic acetylcholine receptors (nAChR) and GABA receptors (GABAR) are expressed
in the lung. Specifically, the nAChR >7 and > 4 subunits and the GABAR
subunits A2 and A3 are important in the lung. Despite the importance of
acetylcholine and GABA signaling in lung morphology and apoptosis,
little is known about the effect of IUGR on these cell signaling systems.
We hypothesize that IUGR will decrease mRNA levels of nicotinic acetylcholine (>4 and >7) and GABA (A3) receptors in the rat lung at birth
(pre-alveolarization), and that this decrease will persist through day 21
(post-alveolarization).
Methods Used: IUGR (induced by bilateral uterine artery ligation) and
control rat lungs were harvested at birth, day 7, and day 21. Real-time
RT-PCR was used to measure the mRNA levels of nAChR subunits >4
and >7 and GABAR subunits A2 and A3.
Summary of Results: Results are IUGR as a % of control. At birth, IUGR
significantly decreased mRNA levels of nAChR >4 (73%*) and >7 (59%*)
relative to control; mRNA levels of GABAR A3 trended lower (79%,
p=0.094) in IUGR rats. At day 21, IUGR significantly increased the expression of nAChR >4 (146%*); the expression of nAChR >7 (128%,
p=0.099) and GABAR A3 (126%, p=0.078) trended higher in IUGR rats.
pe0.001.
Conclusions: We conclude that IUGR affects cholinergeric receptors differently before and after alveolarization. Specifically, IUGR decreases mRNA
levels of two types of nAChR at birth, prior to the onset of alveolarization.
We speculate that the protein levels of both nicotinic acetylcholine receptors will be similarly affected. Interestingly, mice with a knock-out >7
nAChR gene exhibit increased cellular hyperplasia. Therefore, we further
speculate that the decreased expression of the cholinergic receptors play an
important role in the changes in morphology and apoptosis seen in IUGR.
376
MATERNAL TOBACCO SMOKE EXPOSURE ALTERS
RAT LUNG PPARF AND SETd8 ABUNDANCE IN A
GENDER-SPECIFIC MANNER
Joss-Moore L, Wang Y, Hale MA, Campbell M, Fitzhugh M, Kaur R,
Jiang C, Brown A, Yu X, Callaway C, McKnight R, Malleske DT, Fung
C, Albertine K, Lane RH University of Utah, Salt Lake City, UT.
Purpose of Study: Fetal exposure to maternal tobacco smoke (MTS)
produces growth restriction with increased lung disease in infancy and
adulthood. Animal models of MTS show alterations in alveolar development.
Alveolar development depends upon precisely timed regulation of gene expression. Environmental insults, such as MTS exposure, can disrupt gene
expression by modifying epigenetic characteristics of chromatin. One chromatin modifying enzyme is the histone lysine methyltransferase, Setd8.
Transcription of Setd8 is directly regulated by PPARF. PPARF is crucial
for alveolar formation and its levels are decreased by placental insufficiency. Despite these connections, the effect of MTS on lung Setd8 and
PPARF expression in offspring is unknown.We hypothesize that MTS will
decrease mRNA and protein levels of Setd8 and PPARF isoforms in the
rat lung at birth.
Methods Used: Maternal rats were subject to tobacco smoke exposure from
E11 to term, pups were delivered by c-section and lungs removed. Real-time
RT-PCR and immunoblot were used to measure mRNA and protein levels
respectively of Set8 and 2 isoforms of PPARF.
Summary of Results: In female rat lungs, MTS decreases Setd8 mRNA
(85%*) and protein abundance (69%*) relative to control at birth. In contrast, in male rat lungs, MTS increases Setd8 protein abundance (130%*)
relative to control, with no change in Setd8 mRNA levels. MTS also decreases mRNA levels of PPARF in male (62%*) and female lungs (61%*)
with a similar reduction of PPARF protein abundance in male (68%*) and
female (69%*) lungs.*pe0.05.
Conclusions: We conclude that MTS exposure alters rat lung Setd8 expression in a gender specific manner at birth. In female rat lungs, the decrease in Setd8 is consistent with a decrease in PPARF. Interestingly, in
male lungs, a similar reduction in PPARF is associated with an increase
in Setd8 abundance. These data are intriguing and may reflect an increase
in male Setd8 protein stability. We speculate that alterations in Setd8 abundance may affect histone methylation status of genes important for alveolar
formation.
193
Western Regional Meeting Abstracts
Journal of Investigative Medicine
377
MECHANICAL TACTILE STIMULATION IN A NEONATAL
STRESS MODEL ALTERS DEPOT-SPECIFIC ADIPOSE
TISSUE EXPRESSION OF TNF-ALPHA AND IL-6 OF
ADULT RATS
Warner G, Haley S, Gulliver K, Lane RH, Moyer-Mileur LJ University of
Utah, Salt Lake City, UT.
Purpose of Study: Preterm hospitalized infants experience numerous to
stressful events. Early life stress is associated with greater fat mass and
preferential visceral adipose tissue (VAT) deposition. VAT produced inflammatory cytokines, TNF-alpha and IL-6, are linked to obesity and insulin resistance. Infant massage has been shown to decrease stress. Using
mechanical-tactile stimulation (MTS, a surrogate for infant massage), we
tested the hypothesis that MTS administered during neonatal stress would
reduce VAT depot mRNA expression of TNF-alpha and IL-6.
Methods Used: Timed pregnant dams were delivered at term (E21), litters
culled to 10 pups(5 M, 5 F), and randomized to: control (CTL; 60 min
maternal separation), neonatal stress (NS; maternal separation + injection +
hypoxia/hyperoxia) and NS + MTS (10 min of stroking and limb movement).
Treatments were given from D5-9 with tissue harvested on D21 and D120.
Subcutaneous (SAT)/VAT depots were detected by MRI. Serum TNF-alpha
and IL-6 levels were measured with ELISA with VAT/SAT depot TNF-alpha
and IL-6 mRNA levels by real-time PCR.
Summary of Results: At weaning (D21) body weight and SAT/VAT
depots were similar. In adults (D120), MTS decreased VAT depot versus
NS or CTL (pG0.04); body weight and SAT depot were not different. At
weaning VAT depot IL-6 and TNF-alpha expression was higher in males
than females (pG0.05). In adults, VAT depot TNF-alpha mRNA levels were
higher in NS than CTL (pG0.02) with higher IL-6 mRNA expression in
both NS and MTS compared to CTL (pG0.05). SAT depot TNF-alpha and
IL-6mRNA expression and serum TNF-alpha and IL-6 levels were similar
at D21 and D120.
Conclusions: MTS administered during neonatal stress decreased adult
VAT deposition and VAT TNF-alpha mRNA expression. Neonatal stress elevated VAT depot expression for both IL-6 and TNF-alpha mRNA in adult
rats. VAT depot TNF-alpha expression is associated with obesity and reduced
insulin sensitivity. We speculate that MTS attenuates early life stress and
promotes age specific changes to signaling pathways affecting adipose tissue
deposition and inflammatory cytokine expression.
378
INTRAUTERINE GROWTH RESTRICTION ALTERS
EXPRESSION OF PPARF CO-REPRESSORS: Ncor1, SMRT,
AND SIRT1 IN JUVENILE MALE RAT ADIPOSE
Lang B, Wang Y, Yu X, Hale M, Callaway C, McKnight R, Joss-Moore L,
Lane RH University of Utah, Salt Lake City, UT.
Purpose of Study: Intrauterine growth restriction (IUGR) predisposes
individuals to adult onset obesity. In a well characterized rat model of IUGR,
we demonstrated altered adipogenesis in juvenile males with increased visceral adiposity and increased expression of peroxisome proliferator activated
receptor gamma (PPARF). PPARF is a key regulator in adipogenesis. PPARF
activity is inhibited by the binding of a co-repressor complex composed of
nuclear receptor co-repressor 1 (Ncor1), silencing mediator of retinoic acid
and thyroid receptor hormone (SMRT), and NAD+ dependent deacetylase
SIRT1. Increased expression of PPARF causes adipogenesis and weight gain;
however, the expression of Ncor1, SMRT, and SIRT1 in response to altered
adipogenesis in the setting of IUGR remains unknown. We hypothesize that
IUGR will alter mRNA and protein levels of the co-repressor complex
components, NCor1, SMRT, and SIRT1, in the visceral adipose tissue of
juvenile male rats.
Methods Used: To test this hypothesis we used a well characterized UPI
induced IUGR rat model. Expression of Ncor1, SMRT, and SIRT1 mRNA
and protein were measured in subcutaneous (SAT) and visceral adipose tissue
(VAT) at day 7 and day 21.
Summary of Results: In VAT, IUGR increases mRNA levels of SMRT in
day 7 and day 21 male rats (224T72* and 234T14*) and Ncor1 in day 21 male
rats (227T32*) when compared to controls. IUGR increases SIRT1 protein
levels of day 21 male rats (321T69*) with no change in SIRT1 mRNA levels.
In SAT, IUGR increases mRNA levels of Ncor1 and SMRT in day 7 male rats
(164T20* and 180T12*) and SMRT in day 7 female rats (114T10*) when
194
&
Volume 59, Number 1, January 2011
compared to controls. IUGR decreases SIRT1 protein levels of day 21 female
rats (47T10*). (*pG.05).
Conclusions: IUGR increases mRNA levels of NcoR1, SMRT and alters
protein levels of SIRT1, in an age, gender, and adipose depot specific manner.
In metabolic models, gender variation is a persistent observation and males
are often more affected than females. We speculate that this increase in corepressor complex expression in IUGR male rats is an appropriate response
to their increased PPARF expression; however, the complex is ineffective in
inhibiting PPARF activity.
379
TISSUE SPECIFIC ROLE OF DNA METHYLTRANSFERASE
4 DURING ZEBRAFISH DEVELOPMENT
Hooton J1, Jafri I2, Jones DA2 1University of Washington School of
Medicine, Seattle, WA and 2University of Utah, Salt Lake City, UT.
Purpose of Study: DNA methylation is a heritable epigenetic marker. The
mechanism by which DNA methylation and gene expression are regulated
during embryonic development is an area of active research. The zebrafish
presents a unique model which is well suited to investigate mechanisms of
normal embryonic development because of its easily observed rapid embryonic development and availability of tools to manipulate gene expression.
Recent research has started to establish tissue specific roles for the various
DNA methyltransferases (DNMTs) during development. These studies have
also shed some light on the basic mechanisms by which DNMTs regulate
expression of targeted genes. DNMT3 is one of the three groups of DNMTs
found in humans and it has six known zebrafish orthologs: Dnmt3/4/5/6/7/8.
The focus of this project was to investigate the role of zebrafish dnmt4 in
normal development. Whole mount in situ hybridization assays revealed that
dnmt4 is robustly expressed in hematopoietic compartments as well as the
site of hematopoietic stem cell generation. We hypothesized that dnmt4
controls zebrafish hematopoiesis by regulating the transcription of hematopoietic genes.
Methods Used: We took a candidate approach and investigated whether the
knockdown of dnmt4 caused transcriptional up regulation of aid, mbd4, and
gadd45a. These targets were chosen based on recent findings both in published studies and our lab_s unpublished work that indicates they play a role in
hematopoiesis. Antisense morpholino knockdown technology was used to
create zebrafish embryos with substantially decreased dnmt4 expression.
RNA was isolated from whole fish samples and then cDNA was created. RTPCR was conducted to test the relative expression normalized to 28s of the
target genes.
Summary of Results: Expression of our target genes was found to be two
to three times higher in dnmt4 morphants than in controls.
Conclusions: Taken together these data provide evidence that dnmt4 plays
a role in hematopoietic differentiation and establishes a possible tissue specific role for dnmt4 during embryonic development. Future studies will be
aimed at further characterizing the hematopoetic phenotype of dnmt4 morphants and verifying that changes in gene expression correlate with tissue
specific changes in DNA methylation of target genes.
380
MECHANICAL-TACTILE STIMULATION DURING
NEONATAL STRESS DECREASES VISCERAL ADIPOSE
TISSUE AND PREVENTS HYPERINSULINEMIA IN
ADULT RATS
Warner G, Haley S, Gulliver K, Thomson JA, Joss-Moore L, Lane RH,
Moyer-Mileur LJ University of Utah, Salt Lake City, UT.
Purpose of Study: Preterm infants are continually challenged by physiologic and environmental stressors in early life. Early life stress is associated
greater abdominal visceral adipose tissue (VAT). Increased VAT is linked to
adult metabolic consequences such as glucose intolerance and insulin resistance independent of obesity. Infant massage decreases stress biomarkers.
Using mechanical-tactile stimulation (MTS) as a surrogate for infant massage, we tested the hypothesis that MTS administered to rat pups during
neonatal stress would decrease VAT depot and improve circulating levels
of glucose, insulin, and the adipokines leptin and adiponectin.
Methods Used: Timed pregnant dams were delivered at term (E21). Litters
were culled to 10 pups (5 M, 5 F) and divided into 3 groups: naBve control (CTL),
neonatal stress (NS; maternal separation + injection + hypoxia/hyperoxia) and
NS + MTS (10 min of tactile stimulation and limb movement). Treatments
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
were given D6-10 and tissue harvested on D120. Animals fasted for 24 hr prior
to serum collection. Measurements included glucose by glucometer, insulin,
leptin, and adiponectin by ELISA, DXA total fat mass (g), and MRI abdominal
subcutaneous (SAT) and VAT depots. ANCOVA was used to determined significance at pG0.05.
Summary of Results: MTS significantly decreased VAT depot versus
CTL (pG0.01) as well as the VAT:SAT ratio when compared to NS or
CTL (pG0.01). MTS serum leptin was 160% higher than CTL (p=0.01)
despite similar total fat mass and SAT depot. In NS males, fasting insulin
was two-fold higher compared to MTS or CTL males (pG0.001) concurrent
with a 75% reduction in serum adiponectin versus CTL (p=0.03). No differences in fasting glucose levels were detected.
Conclusions: We conclude MTS administered during neonatal stress
decreases VAT depot and increases serum leptin and adiponectin levels in
adult rats. In rats, leptin and adiponectin increase insulin sensitivity. The
hyperinsulinemia observed in NS adult male rats may be due to altered
leptin and adiponectin which decreased adipose tissue sensitivity. We speculate that MTS during neonatal stress attenuates the stress response leading
to changes to signaling pathways affecting adipose tissue deposition and
sensitivity in rats.
381
INTRAUTERINE GROWTH RESTRICTION BY
UTEROPLACENTAL INSUFFICIENCY AND MATERNAL
TOBACCO SMOKING EXPOSURE CAUSES
GENDER-SPECIFIC CHANGES IN MRNA EXPRESSION
OF HIPPOCAMPAL ESTROGEN RECEPTOR
Numpang B, Caprau D, Ke X, Hale M, Yu X, Jiang C, Kaur R, Fitzhugh M,
Callaway C, Joss-Moore L, McKnight R, Lane RH University of Utah,
Salt Lake City, UT.
Purpose of Study: 2 major causes for intrauterine growth restriction
(IUGR) in the United States are uteroplacental insufficiency (UPI) and maternal tobacco smoking exposure (MTSE). Both UPI and MTSE impacts
neurodevelopmental outcomes in offspring born with IUGR, particularly
in males. A common characteristic of IUGR animal models is increased
hippocampal apoptosis. Neuronal apoptosis is modulated by estrogen signalling - estrogen is anti-apoptotic in the hippocampus. Estrogen mediates
it effects on hippocampal apoptosis through estrogen receptors (ER), which
are encoded by the genes esr1 and esr2. Currently, little is known about the
effects of IUGR by either UPI or MTSE on hippocampal expression of ERs.
Methods Used: IUGR was induced by 2 methods in 2 separate experimental groups: (1) uterine ligation in pregnant rats at day 19 of gestation
to mimic UPI, and (2) exposure of pregnant rats to tobacco smoke from day
11 of gestation to term to mimic MTSE. Both groups underwent caesarean
section at term. The pups were killed, hippocampi were dissected and RNA
extracted. Reverse transcriptase real-time PCR was performed to assay expression of esr1 and esr2.
Summary of Results: UPI decreased esr1 (51% reduction, p=0.01) and
esr2 (55% reduction, p=0.02) expression in male pups compared to controls.
UPI did not alter esr1 or esr2 expression in female pups. In contrast, MTSE
increased esr2 expression (123% increase, p=0.05) in female pups compared
to controls, but no changes were seen in males. MTSE did not alter esr1
expression in either gender.
Conclusions: We conclude that the consequence of UPI and MTSE on
hippocampal ER mRNA expression is different, despite both UPI and MTSE
causing IUGR. The mechanisms behind MTSE induced IUGR is unknown,
but the impact is likely through MTSE induced fetal exposure to UPI, nicotine, and/or some other compounds. Given the different observed effects of
MTSE and UPI on hippocampal ER gene expression, we speculate that
mechanisms through which MTSE affects ER expression occur through
exposure to nicotine and/or some other compound.
382
MATERNAL SMOKE EXPOSURE DURING PREGNANCY
DECREASES RAT HEPATIC IGF-1 mRNA VARIANT LEVELS
IN A GENDER SPECIFIC MANNER
Lang B, Wang Y, Jiang C, Kaur R, Brown A, Campbell M, Fitzhugh M,
Hale M, Callaway C, Yu X, Fung C, Malleske DT, McKnight R, Joss-Moore L,
Albertine K, Lane RH University of Utah, Salt Lake City, UT.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Purpose of Study: Maternal cigarette smoking during pregnancy causes
intrauterine growth restriction (IUGR). IUGR is the result of fetal adaptation
to a suboptimal in-utero environment. Insulin like growth factor 1 (IGF-1)
plays a key role in fetal growth and serum levels are directly correlated to
birthweight. Serum IGF-1 levels are controlled by hepatic IGF-1 gene expression. IGF-1 gene expression is characterized by multiple mRNA variants,
which affect IGF-1 translation efficiency via multiple promoters and alternative splicing. The IGF-1 gene and mRNA variants are highly conserved
among mammals. IUGR affects IGF-1 in the setting of perinatal malnutrition
with males more affected than females, however; the effects of IUGR from
maternal cigarette smoke exposure on hepatic IGF-1 mRNA levels remain
unknown. We hypothesize that maternal cigarette smoke exposure (MSE)
during pregnancy will decrease hepatic IGF-1 mRNA variant level (IGF-1A,
IGF-1B, P1, and P2) in the newborn offspring, with males more affected
than females.
Methods Used: Cigarette smoking was initiated on day 11 of gestation
and continued to day 21 when rat pups were delivered via c-section. Liver
harvested at day 0 and mRNA quantified via real time RT-PCR.
Summary of Results: MSE during pregnancy decreased IGF-1
mRNA variants P1 and IGF-1A in day 0 females (62.5%T8* and 73%T6*)
when compared to controls. MSE during pregnancy decreased IGF-1
mRNA variant P1 in day 0 males (70%T6*) when compared to controls.
(*pG0.05).
Conclusions: We conclude that MSE during pregnancy decreases IGF-1
mRNA variant levels similarly in both genders. Gender variation is a persistent observation in metabolic models, and males are often more affected
than females. Uteroplacental insufficiency (UPI) IUGR showed an ongoing
decrease in males IGF-1 expression into adolescence. Thus far, MSE IGF-1
mRNA variant levels are very similar to findings in the UPI IUGR model.
Thus, we speculate the primary fetal effects of MSE are due to placental
insufficiency as opposed to toxin exposure.
383
INTRAUTERINE GROWTH RESTRICTION ALTERS
CARDIAC GENE mRNA LEVELS
Bierer R, Kaza E, Hale M, Yu X, Callaway C, Joss-Moore L, Lane RH
University of Utah Division of Neonatology, Salt Lake City, UT.
Purpose of Study: Intrauterine growth restriction (IUGR) predisposes
individuals to cardiovascular disease in adulthood that results in cardiac
hypertrophy. IUGR also causes cardiac hypertrophy in adult rats. Several
genes are specifically expressed during heart development that are abnormally upregulated in cardiac hypertrophy including ANP, BNP, HCN4,
NR2B, and >-skeletal actin. These genes are all partially regulated by a
common master regulatory complex. However, the effect of IUGR on expression of these genes is unknown. We hypothesize that IUGR will increase
mRNA levels of the genes that code for ANP, BNP, HCN4, NR2B and >-skeletal
actin in rat hearts.
Methods Used: Bilateral uterine artery ligation surgery was performed
on rat dams at E19. Rat pup hearts were harvested at term (D0) and D21.
mRNA was isolated and cDNA was produced. Real-time RT-PCR was performed to assess mRNA levels of Nppa (gene that encodes ANP), Nppb
(BNP), HCN4, Grin2b, (NR2B) and Acta1 (>-skeletal actin).
Summary of Results: Results are presented as IUGR as % of Control T
SEM. IUGR increased mRNA levels of Nppa(306T20%***), HCN4
(167T14%*), and Grin2b (187T19%*) in D21 males. IUGR also increased
mRNA levels of HCN4 (150T12%**) in D21 females. IUGR decreased
mRNA levels of Acta1 (75T2%**) in D21 males. No differences were
seen in Nppb mRNA levels or in any D0 animals. *pe0.05, **pe0.01,
***pe0.001.
Conclusions: We conclude that IUGR increases mRNA levels of Nppa,
HCN4, and Grin2b in male rats, but only HCN4 in female rats. Contrary to
our hypothesis, IUGR decreases Acta1 mRNA levels in male rats. Gene
specific binding with the master transcriptional repressor complex, REST
(repressor element 1-silencing transcriptional factor), may account for this
variability. REST binding involves variable co-repressors that alternately
cause transcriptional expression or repression in response to different stimuli.
We speculate that IUGR will increase protein levels of ANP, HCN4, and
Grin2b in the rat heart. We also speculate that IUGR will alter REST expression and binding.
195
Western Regional Meeting Abstracts
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Our most important findings are: In D0 hippocam384
HEME OXYGENASE-1 EXPRESSION IS DIFFERENTIALLY
REGULATED IN VITRO BY METALLOPORPHYRINS
Schulz-Geske S, Zhao H, Kalish FS, McCarthy E, Vreman HJ, Wong RJ,
Stevenson DK Stanford University School of Medicine, Stanford, CA.
Purpose of Study: Neonatal jaundice is caused by an increased production
of bilirubin and a decreased capacity to eliminate bilirubin. Bilirubin is
produced from heme degradation, mediated by heme oxygenase (HO), the
rate-limiting enzyme. Metalloporphyrins (Mps), synthetic derivatives of
heme, have been shown to be potent competitive inhibitors of HO. However,
some may induce the transcription of HO-1 via the de-repression of Bach1,
a heme-sensitive transcriptional factor that suppresses the HO-1 gene. In this
study, we investigated the regulatory mechanism(s) by which Mps induce
HO-1 expression.
Methods Used: NIH3T3 HO-1-luc cells (HO-1-luc), stably transfected
with the full-length HO-1 promoter fused to the reporter gene luciferase
(luc) were incubated with 15-KM chromium mesoporphyrin (CrMP) or zinc
deuteroporphyrin bis glycol (ZnBG). Single cell HO-1 promoter activation
was monitored in real-time over 48h using Bioluminescence Microscopy
(BLM) and expressed as mean luminescence intensity from 6 regions (cells)
of interest. Cells treated with heme (30 KM), a potent HO-1 inducer, served
as positive controls. HO-1 mRNA was quantitated by qRT-PCR in HO-1-luc
and NIH3T3 cells, stably expressing Bach1 shRNA (shB), after 24h of incubation with 15-KM CrMP or ZnBG. HO-1 and Bach1 proteins were
detected by Western Blot.
Summary of Results: In HO-1-luc cells, peak activation of the HO-1
promoter by heme occurred within 4Y9h, whereas HO-1 promoter activity
peaked periodically over 48h after Mp treatment. Both Mps significantly
increased HO-1 mRNA and protein and decreased Bach1 protein, but to a
much lesser extent after ZnBG treatment (Table). HO-1 mRNA levels in shB
cells were significant lower after CrMP treatment compared to HO-1-luc
cells. In contrast, no difference in HO-1 mRNA levels between HO-1-luc and
shB cells were observed after ZnBG treatment.
Conclusions: HO-1 Induction by CrMP is mediated by the de-repression
of the transcriptional factor Bach1, whereas the effect of ZnBG on Bach1
needs further elucidation. HO-1 protein induction following ZnBG and CrMP
treatment may be mediated by the resultant increase in heme due to the inhibition of HO.
385
MATERNAL TOBACCO SMOKE EXPOSURE INFLUENCES
GENES INVOLVED IN HIPPOCAMPAL DEVELOPMENT IN
A GENE AND GENDER SPECIFIC MANNER
Numpang B1, Caprau D1, Ke X1, Schober M2, Hale M1, Callaway C1, Jiang C1,
Fitzhugh M1, McKnight R1, Joss-Moore L1, Lane RH1 1University of Utah,
Salt Lake City, UT and 2University of Utah, Salt Lake City, UT.
Purpose of Study: Maternal tobacco smoke exposure (MTSE) is detrimental for fetal brain development. In humans and animal models, MTSE
predisposes to postnatal learning disabilities with effects persistent throughout
the life span. Changes in gene expression due to changes in epigenetic markers
are believed to be responsible for the persistence of a phenotype long after an
initial insult. Despite the known detrimental effect of MTSE on the developing
brain, it is unknown how MTSE affects expression of genes involved in hippocampal development and learning processes. We hypothesize that in the
hippocampus of d0 rats, MTSE alters the mRNA expression of genes involved
in hippocampus development, such as insulin like growth factor 1 (IGF1),
NMDA receptors (NMDA-NR1) and glucocorticoid receptor (GR) pathways
genes and variants.
Methods Used: To test our hypothesis we used hippocampal tissue from
Sprague-Dawley rats exposed to cigarette smoke from day 11 of gestation
until term and control rats (n = 12/group). We quantified hippocampal mRNA
expression of genes and gene variants of IGF1, NMDA receptors and GR
pathways.
196
pus MTSE 1) increased mRNA of total IGF1 and variants (to 200-350% of
control levels) in females and increased IGF1A mRNA (to 400% of control
levels) in males. 2) increased mRNA expression of NMDA-NR1 and variants
(to 250-700% of control levels) in females and decreased mRNA of total
NMDA-NR1 in males (to 22% of control levels). 3) decreased mRNA of GR
in males (to 40% of control levels) while no change was seen in females.
Conclusions: Two conclusions arise from this data. First, the fetal hippocampal mRNA levels are exceptionally sensitive to MTSE and the effects are
gene specific. Most of the genes studied present with changes in mRNA
expression following MTSE. Second, the fetal hippocampal response to
MTSE is gender specific. We speculate that upon further testing, we will
continue to identify gene and gender specific differences in hippocampal
protein expression and epigenetic markers, associated with gender specific
behavioral changes later in life.
Neonatal Y Pulmonary II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
386
DECREASED PULMONARY VESSEL DENSITY AND
PULMONARY ARTERY ENDOTHELIAL CELL FUNCTION
IN FETAL SHEEP WITH INTRAUTERINE GROWTH
RESTRICTION
Rozance PJ, Seedorf G, Brown A, Gien J, Tang J, Abman S University of
Colorado, Aurora, CO.
Purpose of Study: Intrauterine growth restriction (IUGR) increases the risk
for neonatal respiratory disease including bronchopulmonary dysplasia and
associated pulmonary vascular dysfunction. Abnormal lung growth and
structure have been noted in animal models of IUGR but whether IUGR
adversely impacts fetal pulmonary vascular development and pulmonary
artery endothelial cell (PAEC) function is unknown. Therefore, we hypothesized that fetal pulmonary alveolarization and vascular growth are decreased
in experimental IUGR and that PAECs from IUGR animals have abnormalities of growth and tube formation in vitro.
Methods Used: Studies were performed in an established model of placental insufficiency and IUGR induced by exposing pregnant ewes to elevated temperatures (day 37Y120; term 148). Late gestation alveolarization
was quantified by radial alveolar counts (RAC) and pulmonary vessel density
by staining for von Willebrand Factor. Proximal PAECs were isolated by
enzymatic digestion and cultured by standard methods. In vitro PAEC
function was tested between passages 4Y6 by measuring growth and tube
formation assays in the presence or absence of insulin (50Y200 nmole/L) or
rhVEGFA (50ng/ml).
Summary of Results: RACs were decreased 20% (5.7T0.3 vs. 7.2T0.2,
pG0.005) and pulmonary vessel density was decreased 44% in IUGR
(5.3T0.6 vs. 9.5T0.9 vessels/hpf, PG0.01). In normal PAECs, tube formation
as assessed by branch point counting was increased to a maximum of 39% in
a dose response fashion with insulin treatment (pG0.0001) and by 48% with
VEGFA treatment (pG0.0001). In comparison with normal PAECs, tube
formation by IUGR PAECs at baseline was reduced by 34% and was not
responsive to treatment with insulin or VEGFA (pG0.0001). In normal
PAECs, insulin increased cell growth to a maximum of 68% in a dose response fashion (pG0.01). Cell growth was reduced in IUGR PAECs by 29% at
baseline (pG0.05) and the response to insulin was also attenuated (pG0.01).
Conclusions: Alveolarization, lung vascularity, and in vitro PAEC function
are impaired in IUGR fetuses. We speculate that the pulmonary circulation
of IUGR fetuses is characterized by endothelial dysfunction, which may
contribute to abnormal growth, development, and the increased risk for
chronic lung disease and bronchopulmonary dysplasia in IUGR.
387
IUGR INCREASES MECP2 MRNA AND CALPONIN
ABUNDANCE IN NEONATAL RAT LUNG EXPOSED TO
MODERATE HYPEROXIA
Sainz A, Stiers J, Wang Y, Yu X, Callaway C, McKnight R, O’Brien E,
Albertine K, Lane RH, Joss-Moore L University of Utah, Salt Lake City, UT.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Purpose of Study: Intrauterine growth restriction (IUGR) alters lung development and increases the risk of bronchopulmonary displaysia (BPD),
with male infants more affected. IUGR infants who develop BPD are often
subject to a secondary insult, such as hyperoxia. In animal models, neonatal
hyperoxia exposure leads to increased lung fibrosis, a characteristic of BPD.
Fibrosis is associated with increases in the methyl CpG binding protein,
MeCP2. However, it is unknown if IUGR increases the effects of neonatal
hyperoxia on MeCp2 or calponin (a marker of fibrosis) in the rat lung. We
hypothesize that male IUGR rat lungs will be more susceptible to moderate
neonatal hyperoxia and will display increased MeCp2 mRNA levels and
calponin abundance compared to hyperoxic control rats.
Methods Used: IUGR (induced by uterine ligation) and control rat pups
were exposed to 60% oxygen or room air from postnatal days 3-6 (beginning
alveolar formation), then room air only to postnatal day 21 (completion
of alveolar formation). MeCp2 mRNA and calponin protein was quantified
in day 21 lungs in 4 groups; normoxic control (NC), normoxic IUGR (N-IUGR),
hyperoxia control (HC) and hyperoxia IUGR (H-IUGR).
Summary of Results: N-IUGR increases MeCP2 mRNA in female lungs
(132%*) relative to NC, with no change in male MeCP2 mRNA levels or
calponin abundance in either gender. H-IUGR increased MeCp2 mRNA
in both male (125%*) and female (150%*) lung relative to HC. In contrast,
H-IUGR increased calponin abundance only in male lung (145%) relative to
HC. In female lung, H-IUGR decreased calponin levels relative to HC
(73%*). *pe0.01.
Conclusions: We conclude that IUGR increases the effects of neonatal
hyperoxia on lung MeCP2 mRNA and calponin abundance in male rats.
However, in female rats, IUGR decreases the effects of hyperoxia on calponin
abundance. We speculate that the male IUGR lung may be more susceptible
to the development of fibrosis as a result of a secondary insult such as
hyperoxia.
388
A NOVEL MEANS FOR DELIVERING NASAL
INTERMITTENT POSITIVE PRESURE VENTILATION
(NIPPV) IN INFANTS VIA THE NASAL CANNULA (NC):
MEASUREMENTS OF DELIVERED PARAMETERS
Andaya SC1, Crotwell D2, DiBlasi R2, Ramanathan R1 1Keck School of
Medicine of USC, LAC+USC Medical Center, Los Angeles, CA and 2Seattle
Children’s Hospital Research Institute, Seattle, WA.
Purpose of Study: Nasal ventilation using NIPPV is on the rise in preterm
infants to decrease extubation failures, bronchopulmonary dysplasia, and for
the treatment of apnea of prematurity. NIPPV is typically applied using
continuous positive airway pressure (CPAP) bi-nasal short prongs which can
result in nasal trauma. Nasal Cannula Intermittent Mandatory Ventilation
(NC-IMV) is a novel means of delivering pressure controlled NIPPV breaths
noninvasively to neonates requiring respiratory support. We previously
reported that NC-IMV is feasible and well tolerated in a large number of
neonates. However, pressures/volume delivered to the patient is not known.
Our goal is to determine the magnitude of pressure, volume, and positive endexpiratory pressure (PEEP) delivered to an infant nasal airway/lung model
using different sized NC at different peak inspiratory pressure (PIP) settings
during time-cycled, pressure-limited mode.
Methods Used: We configured a neonatal test lung to simulate an apneic
premature infant with compliance 0.8 mL/cmH20, resistance 75 cmH20/L/
sec. An infant nasal airway model was attached to the test lung. The ventilator
set in IMV mode, rate 40 breaths/min, inspiratory time 0.5 s, Flow 7Y9 L/min,
and PEEP 5 cmH20. The nasal airway was ventilated at PIP of 10, 15, 20, 25,
and 30 cmH20 using infant and intermediate high-flow NC (Fisher Paykel,
Auckland, NZ) and a new prototype NC (Neotech Nasal Cannula\). Pressure, volume, and PEEP were measured in the test lung.
Summary of Results: Under all testing conditions, there was detectable
pressure, volume and PEEP during NC-IMV. There was a linear relationship
between PIP applied by the ventilator and volume up to 30 cmH20. The
Neotech Nasal Cannula\ provided greater pressure, volume, and PEEP than
the other infant NC.
Conclusions: NC-IMV can provide a significant amount of a neonate’s
volume/pressure requirements with less nasal trauma than CPAP prongs.
Further studies are underway to evaluate the pressure/volume deliveries and
reduction of nasal injury in spontaneously breathing neonates on NC-IMV
support.
* 2011 The American Federation for Medical Research
389
PROTEIN AND CATION CONCENTRATIONS IN THE
EXHALED BREATH FLUID OF PREMATURE NEONATES
Morales EM Harbor UCLA, Torrance, CA.
Purpose of Study: Ventilator associated pneumonia (VAP) and pulmonary
edema are common events in premature neonates. Airway fluid can be collected from these babies for diagnostic and investigative studies by lavaging
the lungs with saline, but this involves some risk in terms of gas exchange and
infection. Alternatively, respiratory fluid and solutes can be collected from
filters in the expiratory tubing of the ventilator. This noninvasive approach
can be repeated without endangering the child. We hypothesize that data
obtained from expiratory collections can be used to estimate airway solute
concentrations and eventually, early detection and quantification of pathogens
in airway fluid.
Methods Used: Exhaled breath fluid (EBF) samples were collected from the
expiratory filters of conventional ventilators from 4-6 hours of ventilation.
The filters were washed with 3mL of water and measurements were made of
protein, Na+,NH4+, K+, Ca+2 and Mg+2 concentrations in these samples.
(6 samples with disproportionately high Na+ levels due to saline lavage were
excluded). Protein concentration was assayed with a modified colorimetric
(BCA) assay. Cation concentrations were assayed by ion chromatography.
Summary of Results: Protein concentrations (mg/L) averaged 6.0T 6.2
(SD, n=10) EBF samples. Ion concentrations (micromol/L) averaged: Na=
119.7T112.0, K= 5.9T 7.0, Ca=11.8 T11.4, Mg= 2.7T7.9, and NH4+= 0. If it
is assumed that the sum of Na+ and K+ in airway lining fluid (ALF) equals
that in the plasma (assumed to be 150 mEq/L), then dilution (D) of the EBF
by water vapor, (released as a gas from airway surfaces and delivered by
humidifier) averaged 2,355 T 2,140 (n=10). Protein concentrations in the
ALF averaged 14g/L (estimated estimated from the product of average values
of D and EBF filter concentrations). Protein concentrations measured after
washing the airways with 3 mL of saline in 6 neonates averaged 1.8 g/L.
Conclusions: These observations suggest that the ALF is diluted by a factor
of nearly 10 during conventional airway lavage. These studies suggest that
measurements of solutes in EBF may provide a simple and safe approach for
following abnormalities in the airway fluid.
[Supported partially by grants from the NIH (HL-55268, HL-075405,
HD058948, and HD-051857) and the TRDRP (14RT-0073, 15IT-0250,
17RT-0170)].
390
ANALYSIS OF ALVEOLAR MACROPHAGE POLARIZATION
FOLLOWING LUNG INJURY IN PREMATURE NEONATES
Chan B1, Li A1, George R1, Groffen J1, Heisterkamp N1, Kwong K2,
Ramanathan R1, Minoo P1 1University of Southern California, Los Angeles,
CA and 2Harbor-UCLA Medical Center, Los Angeles, CA.
Purpose of Study: Alveolar Macrophages (AM) exhibit significant phenotypic plasticity. Depending on the microenvironment, they undergo distinct
polarization that either propagate inflammation, or alternatively promote
tissue repair. Inflammation related genes are increased in M1-polarized cells;
whereas, M2 polarized cells are important for tissue remodeling. No information is currently available on AM polarization in premature neonates at risk
of Bronchopulmonary Dysplasia (BPD). The imbalance of pro-inflammatory
and anti-inflammatory factors leads to BPD in premature neonates; we speculate
AM polarization may be perturbed in this paradigm. We examined polarization
state of AM obtained from the tracheal aspirates (TA) of preterm neonates at
risk for BPD.
Methods Used: We obtained serial TA samples from a cohort of intubated
premature infants. The neonates were in two groups: Group 1, infants born
between 22 and 30 weeks gestation (High BPD risk). Group 2 included
infants born between 31 and 36 weeks gestation (Low BPD Risk). The inflammatory cells were isolated and subjected to Fluorescence Activated Cell
Sorting (FACS) using antibodies against specific surface markers. Anti-CD14
and Anti-CD163 were used to identify M1- and M2- polarized AM respectively. The patients_ clinical data were also analyzed.
Summary of Results: TA samples from 12 premature neonates were collected and analyzed. Among the subjects, only 5 neonates survived without
BPD. FACS analysis was performed on 47 samples. The percentile of CD14positive AM was highly variable in both High and Low BPD Risk groups.
No differences were found between the two groups. The abundance of
197
Western Regional Meeting Abstracts
Journal of Investigative Medicine
M1-polarized AM did not correlate with the risk for BPD or death.
Interestingly in a small number of Group 2 neonates M2-polarized AM
increased as a function of postnatal age.
Conclusions: To our knowledge, this is the first study to analyze polarization of AM in the lungs of neonates at risk for BPD. The relationship between
AM polarization and BPD appears complex. In the neonates examined, M1
polarization did not correlate with BPD risk. Additional studies are underway
to elucidate the role of AM polarization in BPD.
391
CAFFEINE WORSENS INFLAMMATION AND ALVEOLAR
HYPOPLASIA IN HYPEROXIA-EXPOSED NEWBORN MICE
Lopez B1, Maisonet TM1, Dayanim S1, Grewal S1, Aghai ZH2, Londhe VA1
1
UCLA, Los Angeles, CA and 2UMDNJ, Camden, NJ.
Purpose of Study: To determine the impact of daily caffeine administration on alveolar development in normoxia and hyperoxia. Caffeine therapy
is a current mainstay of NICU management to treat apnea of prematurity.
Although caffeine has also been shown to reduce the incidence of BPD, the
precise mechanism of action remains poorly understood.
Methods Used: Newborn mice from timed-pregnant dams were divided
into 4 experimental groups: 1) saline normoxia (FiO2 21%); 2) caffeine
normoxia; 3) saline hyperoxia (FiO2 80%); and 4) caffeine hyperoxia.
Starting at P1, pups were treated with daily i.p. injection of saline or caffeine
(20 mg/kg load, 10 mg/kg daily caffeine citrate) under constant normoxia or
hyperoxia conditions. Animals were sacrificed at P3 or P15 to collect BAL
and lung tissue for mRNA, protein, and histological analysis.
Summary of Results: Caffeine-treated pups showed poorer weight gain
compared to saline controls. At P3, caffeine-treated lungs showed increased
neutrophil infiltration in normoxia (p = 0.013) and hyperoxia (p = 0.002). Lung
histology at P3 also exhibited larger saccular airspaces in caffeine groups when
compared to saline controls. At P15, morphometric analysis showed fewer
alveoli in caffeine-treated pups exposed to hyperoxia (p = 0.006).
Conclusions: Daily caffeine exposure increases lung inflammation and inhibits alveolar development under normoxia and hyperoxia in newborn mice.
Caffeine is known to inhibit adenosine receptor activation and thereby blocks
down-regulation of inflammation in response to injury. Caffeine may also induce epigenetic changes that alter transcription of critical lung morphoregulatory genes during alveolar development. Our findings suggest that caffeine
may adversely impact alveolar development under hyperoxic conditions.
392
EXPRESSION OF CARCINOEMBRYONIC ANTIGEN CELL
ADHESION MOLECULE 6 (CEACAM6) IN HEALTHY
AND INJURED CEABAC TRANSGENIC MICE
Barrette A1, Lin S2, Chapin C1, Ballard P1 1UC San Francisco, San Francisco,
CA and 2UC San Francisco, San Francisco, CA.
Purpose of Study: CEACAM6 is a glycosylated, GPI-anchored protein
expressed in epithelial cells of many human tissues. It is overexpressed in
Crohn_s disease, facilitating bacterial colonization, and in tumors where it
inhibits apoptosis and promotes metastasis. Expression is hormonally regulated in developing human lung where the protein is localized to plasma
membrane and associates with surfactant (Kolla Am J Physiol Lung 2009).
Using a novel transgenic mouse model with a human bacterial artificial
chromosome containing members of the human CEA family gene cluster
(CEABAC) developed by Chan and Stanners (Molec Therapy 2004), we were
able to examine the role of CEACAM6 in vivo.
The aim of this study was to characterize the expression of CEACAM6 in
the CEABAC mouse lung and explore effects of injury.
Methods Used: Lung tissue and bronchoalveolar lavage (BAL) were collected from wild-type and heterozygous CEABAC mice for RNA and protein
analysis. Large aggregate surfactant was isolated from BAL by centrifugation.
CEACAM6 was measured by Western and immunodot blot assays and normalized to total protein. Lungs were inflation fixed for immunohistochemistry
(IHC). Acute lung injury was induced by intratracheal injection of bleomycin.
Summary of Results: Expression of human CEACAM3, 5, 6, and 7 in the
CEABAC mouse lung was confirmed by RT-PCR. Content of CEACAM6
protein in CEABAC lungs was comparable to that in normal human infant
lung. In BAL, CEACAM6 was localized to the surfactant pellet, and levels
were È10-fold less than in intubated premature infants. By IHC, CEACAM6
was detected in some alveolar epithelial cells (e10%) and in airway epithe-
198
&
Volume 59, Number 1, January 2011
lium. Bleomycin-injured CEABAC mice exhibited weight loss (up to 25%),
but no mortality over treatment periods of up to 19 days. CEACAM6 content
in lung tissue and BAL increased È2-fold over uninjured and saline-treated
mice. IHC revealed markedly increased intensity and number of CEACAM6positive alveolar cells at sites of injury.
Conclusions: We conclude that expression of CEACAM6 in the CEABAC
mouse lung is comparable to that in the human premature infant lung.
CEACAM6 is up-regulated with lung injury and may play a role in cell
proliferation and/or surfactant stabilization under these conditions.
393
CURCUMIN AUGMENTS NEONATAL LUNG
INJURY/REPAIR BY INHIBITING TGF-A SIGNALING
Sakurai R, Li Y, Torday J, Rehan V Los Angeles Biomedical Institute,
Torrance, CA.
Purpose of Study: There is no effective intervention to prevent or treat
BPD. Curcumin has potent antioxidant and anti-inflammatory properties, and
it modulates PPARF signaling, a key molecule in the pathobiology of BPD.
However, its role in the prevention of BPD is not known. The purpose of this
study is to determine if curcumin i) enhances neonatal lung maturation; ii)
protects against hyperoxia-induced neonatal lung injury; iii) protection is
mediated by blocking TGF-A.
Methods Used: Embyronic day (e) 19 fetal rat lung fibroblasts (FRLF) were
exposed to 21% or 95% O2 for 24h following 1h pretreatment with curcumin.
Cell proliferation (thymidine incorporation), differentiation (Western for parathyroid hormone-related protein receptor, PPARF and adipocyte differentiationrelated protein, (ADRP); triolein uptake) and TGFA signaling (Western blot
analysis and confocal immunofluoresce staining for Smad3) were determined.
In a separate set of experiments, neonatal Sprague Dawley rat pups were categorized in the following groups: normoxia (21% O2), normoxia + curcumin
(5 mg/kg BW), hyperoxia (7d exposure to 95% O2), and hyperoxia + curcumin
(5 mg/kg BW). Curcumin was administered i.p. once daily. At the end of the
experimental period, the pups were sacrificed and the lungs analyzed for markers
of lung injury/repair (Western analysis and lung morphology).
Summary of Results: In vitro, curcumin dose-dependently accelerated e19
fibroblast differentiation (increased PTHrP receptor, PPARF, and ADRP
expression and triolein uptake) and proliferation (increased thymidine incorporation). Furthermore, pretreatment with curcumin blocked the hyperoxiainduced decrease (PPARF and ADRP) and increase (>-SMA and fibronectin)
in markers of lung injury/repair. Similarly, in vivo hyperoxia-induced changes
in molecular markers of lung injury repair [PTHrP receptor, PPARF, ADRP,
fibronectin, TGF-A receptor (ALK5) and Smad3 activation] and lung
morphology (radial alveolar count and septal thickness) were effectively
blocked in the curcumin treated group.
Conclusions: Curcumin accelerates lung maturation by stimulating key alveolar epithelial-mesenchymal interactions and blocks hyperoxia-induced
neonatal lung injury by blocking TGF-A activation, suggesting it as a potential
intervention against BPD. (Grant support: NIH (HL-55268, HL-075405,
HD058948, and HD-051857).
394
BDNF PROTEIN ABUNDANCE IN WHITE MATTER IS
DECREASED IN PRETERM LAMBS MANAGED BY
MECHANICAL VENTILATION COMPARED TO NASAL
VENTILATION
Houston B1, Claar J2, Alvord J1, Schober M1, Dong L1, Dahl M1, Block C1,
McKnight R1, Null D1, Yoder BA1, Lane RH1, Albertine K1 1University of
Utah, Salt Lake City, UT and 2Lawrence University, Appleton, WI.
Purpose of Study: Prolonged mechanical ventilation (MV) of preterm neonates is associated with white matter loss and subsequent neurodevelopmental
delays or deficiencies. The molecular basis of these co-morbidities is unknown.
Brain derived neurotrophic factor (BDNF), an endogenously produced antiapoptotic neurotrophin, is important for brain development. The action of
BDNF is counteracted by pro-BDNF, which promotes neuronal apoptosis.
Whether MV affects BDNF and pro-BDNF protein abundance in the brain of
chronically ventilated preterm neonates is not known.We hypothesized that
prolonged MV will lead to less BDNF and more pro-BDNF protein abundance
in white matter of preterm lambs compared to high-frequency nasal ventilation
(HFNV; positive-outcome gold-standard for alveolar formation; similar to
bubble nasal CPAP).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Methods Used: Preterm lambs (È131d gestation; term È150d), treated
Purpose of Study: Sildenafil use is increasing in the management of
with antenatal steroids and postnatal surfactant, were managed by MV or
HFNV for 3d or 21d (n=4/group). Gestation references were for the time of
preterm fetal delivery (F131) and term gestation (T). Temporal lobe white
matter was analyzed by immunoblot (normalized using MemCode kit).
Summary of Results: BDNF protein relative abundance at 3d or 21d of
MV was less than HFNV at the respective times (table; *pG0.05). Pro-BDNF
protein abundance, by comparison, was greater in both groups of preterm
lambs (table; †pG0.05 compared to T).
Conclusions: Preterm birth followed by prolonged ventilation affect the
balance of BDNF and pro-BDNF protein abundance in white matter. This
shifted balance may contribute to more apoptosis in white matter that occurs
in the brain of chronically ventilated preterm lambs (J Invest Med 57:141,
2010). (HL62875, HL56401, HD41075).
persistent pulmonary hypertension (PPHN) of the neonate. There are concerns that sildenafil may contribute to hemodynamic instability in these
neonates. The study objective was to evaluate hemodynamic changes with
sildenafil use in newborns with meconium aspiration syndrome (MAS) or
sepsis and PPHN.
Methods Used: Late preterm and term infants with MAS or sepsis and
PPHN admitted to CHLA NICU between 2004 and 2010 were identified.
Patients with chromosomal abnormalities and congenital heart defects were
excluded. Data on sildenafil dosing, hourly mean blood pressure (MBP), heart
rate (HR), vasopressor/inotrope use were collected for 72 hours after initiation of
po sildenafil. Data were compared between low-dose (G3mg/kg/day) vs. highdose (Q3mg/kg/day) and early (G7 postnatal days) vs. late initiation (Q7 postnatal
days) of sildenafil administration.
Summary of Results: Seventeen patients (12 MAS, 5 sepsis) were identified
with mean gestational age (GA) of 39 T 2 wks, birth weight (BW) of 3191 T
423 g, median APGAR scores at 1, 5, and 10 minutes of 4, 7, and 7 respectively,
sildenafil dose of 2.7 T1.7 mg/kg/day, day of initiation on 8 T 7 postnatal days,
and hours on inhaled nitric oxide (iNO) of 163 T 95 hrs. Ten patients received
low-dose and 7 received high-dose sildenafil. Eight and 9 patients were started
on sildenafil early and late, respectively. There were no differences in GA, BW,
APGAR scores, hours on iNO, or mortality between the groups. After adjusting
for vasopressor/inotrope use during the first 72 hours of sildenafil exposure,
there were no differences from baseline in MBP and HR when comparing
patients on low-dose and high-dose sildenafil. MBPs were similar between
the early and late initiation group; however an increase in HR was seen in the
late group.
Conclusions: In summary, sildenafil administration in neonates with MAS or
sepsis for the management of PPHN was not associated with clinically relevant
changes in MBP or HR after adjusting for vasopressor/inotropic support.
395
MATERNAL FOOD RESTRICTION ASSOCIATED
INCREASE IN LUNG ELASTIN IS DUE TO
PROTEIN DEFICIENCY
Lee R, Sakurai R, Li Y, Satyadev N, Torday J, Rehan V Los Angeles
Biomedical Institute, Torrance, CA.
Purpose of Study: Using a rat model of 50% maternal food restriction
(MFR) during gestation, we have reported altered Blung lipid-differentiation[
programming, explaining the altered pulmonary structure and function in the
affected offspring. Since lung fibroblasts are the key cells that drive lung
lipid-differentiation program and synthesize elastin, a major matrix protein
that determines the lung structure and function, we determined the effect of
MFR restriction on lung elastin gene expression. We also aimed to determine
the specific nutrient component, the deficiency of which leads to altered
elastin expression in the MFR offspring lung.
Methods Used: Pregnant dams 10 days into pregnancy were either
allowed ad lib feeds or given food-restricted diet, i.e., 50% of rat chow
consumed by control dams. After delivery, the pups were sacrificed at
postnatal day (PND) 21 and 9M, when lungs were examined for elastin
gene expression by RT-PCR and Western blotting. To determine the
specific nutrient deficiency that alters elastin gene expression, primary rat
lung fibroblasts were isolated. At 80-90% confluence, the cells cultured
in DMEM for 24h under the following conditions: No fetal bovine serum
(FBS); 1, 5, or 10% FBS; No FBS+Vitamin D (VD, 10-6M); No FBS+
arachidonic acid (ARA, 10-6M)+doxohexanoic acid (DHA, 10-6M); or
No FBS+1, 5, 10% fetal bovine albumin (FBA), following which the
expression of elastin-related genes was determined.
Summary of Results: Compared to ad lib controls, with MFR, at PND 21
and at 9M, there was significantly increased elastin gene expression, at both
mRNA and protein levels (pG0.05, n=4). Culturing primary rat lung fibroblasts
without FBS significantly increased the expression and protein levels of elastin
and the expression of various elastin-related genes such as Lox, FGFR 3 and 4,
and fibrillin 5, while the expression of fibrillin 2 was significantly decreased (all
pG0.05, n=3). Furthermore, while FBS-restriction associated increase in elastin
expression was only partially blocked by either VD or ARA+DHA, it was
completely blocked with 10% FBA supplementation.
Conclusions: Elastin is significantly increased in the lung of MFR offspring
and this increase appears to be largely due to protein deficiency rather than
due to VD, ARA, or DHA deficiency (Grant support-HD58948).
396
SILDENAFIL USE IN NEONATES WITH MECONIUM
ASPIRATION OR SEPSIS: A RETROSPECTIVE LOOK AT
HEMODYNAMIC EFFECTS
Limjoco JJ, Paquette L, Ramanathan R, Seri I, Friedlich P Center for Fetal
and Neonatal Medicine, USC Division of Neonatology, Children’s Hospital
Los Angeles and LAC+USC Medical Center; Keck School of Medicine,
University of Southern California, Los Angeles, CA.
* 2011 The American Federation for Medical Research
397
SILDENAFIL USE IN NEONATES WITH ISOLATED
CONGENITAL DIAPHRAGMATIC HERNIA: A
RETROSPECTIVE LOOK AT HEMODYNAMIC EFFECTS
Limjoco JJ, Paquette L, Ramanathan R, Seri I, Friedlich P Center for Fetal
and Neonatal Medicine, USC Division of Neonatal Medicine Children’s
Hospital Los Angeles and LAC+USC Medical Center; Keck School of
Medicine, University of Southern California, Los Angeles, CA.
Purpose of Study: Sildenafil use is increasing in the management of
persistent pulmonary hypertension (PPHN) of the neonate. There are
concerns that sildenafil may contribute to hemodynamic instability in these
neonates. The study objective was to evaluate hemodynamic changes with
sildenafil use in newborns with congenital diaphragmatic hernia (CDH)
and PPHN.
Methods Used: Late preterm and term infants with isolated CDH admitted
to CHLA NICU between 2004 and 2010 were identified. Patients with
chromosomal abnormalities and congenital heart defects were excluded.
Data on sildenafil dosing, hourly mean blood pressure (MBP), heart rate
(HR), vasopressor/inotrope use were collected for 72 hours after initiation
of po sildenafil. Data were compared between low-dose (G3mg/kg/day) vs.
high-dose (Q3mg/kg/day) and early (G7 postnatal days) vs. late initiation
(Q7 postnatal days) of sildenafil administration.
Summary of Results: Sixteen patients (14 left, 2 right CDH) were
identified with mean gestational age (GA) of 38 T1 wks, birth weight (BW)
of 3343 T 497g, median APGAR scores at 1, 5, and 10 minutes of 5, 6, and
6 respectively, sildenafil dose of 3.8 T1.7 mg/kg/day, day of initiation
on 12 T 8 postnatal days, and hours on inhaled nitric oxide (iNO) of 442 T
286 hrs. Five patients received low-dose and 11 high-dose sildenafil. Five
and 11 patients were started on sildenafil early and late, respectively. No
differences in GA, BW, APGAR scores, or hours on iNO between groups
existed. After adjusting for vasopressor/inotrope use during the first 72
hours of sildenafil exposure, there were no differences in MBP and HR
when comparing patients on low-dose and high-dose sildenafil. MBPs were
similar between the early and late initiation group; however there was an
increase in HR in the late group.
Conclusions: In summary, sildenafil administration in neonates with
CDH for the management of PPHN was not associated with clinically
relevant changes in MBP or HR after adjusting for vasopressor/inotropic
support.
199
Western Regional Meeting Abstracts
Journal of Investigative Medicine
Neuroscience II
Concurrent Session
9:00 AM
Saturday, January 29, 2011
398
MULTIMODALITY IMAGING OF MALFORMATIONS
OF CORTICAL DEVELOPMENT
Behdinian K David Geffen School of Medicine at UCLA, Los Angeles, CA.
Purpose of Study: Focal cortical dysplasia (FCD), Tuberous Sclerosis
Complex (TSC), and Hemimegalencephaly are malformations of cortical development (MCDs) caused by abnormal neuronal proliferation. These MCDs
are often associated with early onset of seizures. Advances in imaging of these
MCDs have improved the sensitivity and specificity of diagnosing subtle MCDs
that are cryptic or hard to identify by MRI. PET/MRI coregistration and magnetoencephalography (MEG) considerably improve the localization of subtle
abnormalities and Diffusion Tensor Imaging (DTI) provides information about
the integrity of white matter adjacent to the lesion. It is shown that using MEG,
DTI, and PET/MRI coregistration provides a non-invasive and more accurate
method of diagnosing and localizing subtle MCDs which in turn results in better
post operative seizure freedom for the patients.
Methods Used: 18-flourodeoxy glucose PET data was coregistered with 3D
MRI using the commercially available Bfusion[ program. Fractional anisotropy(FA) and Apparent diffusion coefficient(ADC) was obtained with the
program BDTI Studio[. Also, in a patient with TSC, DTI tractography map
was obtained.
Summary of Results: PET/MRI coregistration and MEG localized the
epileptogenic in a patient with FCD and subtle MRI abnormalities. Using
PET/MRI coregistration, widespread hypometabolism was shown in the left
hemisphere of a patient with HME who had subtle abnormalities on MRI. In a
patient with TSC, ADC and FA were measured in the area immediately
adjacent to the tuber with the largest associated hypometabolism as shown on
PET/MRI coregistration. ADC=0.86 was higher than the contralateral side
ADC=0.83. FA was 0.17 on the lesional side and 0.27 on the normal side.
Conclusions: Multi-modality imaging of MCDs using PET/MRI coregistration, MEG, and DTI in addition to high quality MRI considerably
improves the ability to non-invasively localize and characterize the epileptogenic lesions when the MRI abnormalities are subtle.
399
SUBCLINICAL MOOD AND STRESS SYMPTOMS
CORRELATED WITH REDUCED CARDIOVAGAL ACTIVITY
Hameed SR1, Stains J2, Ebrat B2, Suyenobu B2, Tillisch K2 1University of
California, Los Angeles, Los Angeles, CA and 2University of California, Los
Angeles, Los Angeles, CA.
Purpose of Study: Autonomic nervous system (ANS) dysfunction is associated with chronic medical disorders such as irritable bowel syndrome,
fibromyalgia, and depression, for which life stress is a risk factor. Whether
ANS dysfunction is a predisposing factor or a consequence of the clinical
disorders is unknown. We propose to examine the relationship between ANS
function, stress, body symptoms, personality traits, and subclinical psychological symptoms in healthy women. We hypothesize that perceived stress,
body symptoms, trait measures, and psychological state symptoms will
correlate with increased sympathetic and decreased cardiovagal activity.
Methods Used: Healthy women aged 18Y50 completed the NEO Personality
Inventory, Hospital Anxiety and Depression (HAD) Questionnaire, Patient
Health Questionnaire (PHQ), State-Trait Anxiety Inventory (STAI), and the
Perceived Stress Scale (PSS). Resting electrocardiogram was acquired and
processed for heart rate variability (HRV) with Kubios software. High Frequency (HF) power represents cardiovagal tone and Low Frequency/High
Frequency (LF/HF) ratio represents sympathetic function. Pearson correlations,
ANOVA, and regression analysis were performed using SPSS 7 software.
Summary of Results: 41 women had complete data. HF correlated with
HAD Anxiety (r=j0.337, p=0.031), HAD Depression (r=j0.390, p=0.012),
and PSS (r=j0.326, p=0.037) but not PHQ, STAI, or NEO scores. No
correlations were found for LF/HF. Using the correlated variables as predictors for HF in a stepwise regression, the only significant predictor was
HAD Depression (R=0.390, B=j0.818, p=0.012).
200
&
Volume 59, Number 1, January 2011
Conclusions: These results suggest that subclinical mood symptoms and
stress but not body symptoms, are linked with cardiovagal activity in healthy
women. Depressive symptoms are the main predictor of decreased cardiovagal activity.
400
PERIPHERAL BLOOD LYMPHOCYTE IMMUNE
CORRELATES IN GLIOBLASTOMA PATIENTS
UNDERGOING DENDRITIC CELL VACCINE
IMMUNOTHERAPY
Fong BM1, Lisiero D1,2, Odesa S1,2, Prins R1,2,3, Liau LM1,2,3 1David Geffen
School of Medicine at UCLA, Los Angeles, CA; 2David Geffen School of
Medicine at UCLA, Los Angeles, CA and 3David Geffen School of Medicine
at UCLA, Los Angeles, CA.
Purpose of Study: Glioblastoma is the most common and malignant primary brain cancer with a median survival of less than 1 year. At UCLA, we
completed a phase 1, dose escalation study to test the safety and feasibility of
autologous tumor lysate pulsed DC vaccination in malignant glioma patients.
To analyze the frequency and activation status of peripheral blood lymphocyte (PBL) population in glioblastoma patients treated with anti tumor lysate
pulsed DC vaccines. Data was then tested to see if a correlation with survival
existed.
Methods Used: 26 patients with glioblastoma were enrolled in this
study. In 13 patients, PBL from pre and post treatment, were stained
with fluorescently-conjugated antibodies (CD3, CD4, CD8, CD16, CD19,
CD25, CD127, CD69) and the composition of lymphocyte subsets and activation status were analyzed using multi-color flow cytometry. The lymphocyte subsets tested for include CD3+CD4+ Helper T cells, CD3+CD8+
cytotoxic T cells, CD3-CD16+ classical Natural Killer (NK) cells, CD3+
CD16+ NKT cells, CD3-CD19+ B cells. The activation status on these
PBMC were simultaneously evaluated by measuring the median fluorescence
intensity (MFI) of CD25 (High affinity IL-2 receptor) and CD69 (early
lymphocyte activation marker).
Summary of Results: Initial data suggests that the degree of increase in
NKT (P value=.094, r2=.662) and CD8 (P value=.240, r2=.414) cell populations post DC+TLR vaccination may correlate with increased survival.
Furthermore, there was a direct correlation with NK cell CD69 activation and
survival (P value=.022, r2=.432). In most patients, differences among CD25
and CD69 values were not statistically significant.
Conclusions: Decreased post vaccination frequencies of NKT and CD8 cell
populations, along with decreased NK cell activation, were associated with
shorter survival. Additional testing is needed to confirm these early findings.
Further study of flow cytometry analyses of glioblastoma patient PBMC
subsets as a monitoring assay to correlate immune response with survival is
warranted.
401
HOW GENOTYPE INFLUENCES CLINICAL FEATURES,
CELLULAR ELECTROPHYSIOLOGY, AND
MICROANATOMY IN PEDIATRIC PATIENTS WITH
TUBEROUS SCLEROSIS COMPLEX
Oh T1, Mathern G1, Kwiatkowski D2, Vinters H1, Salamon N1, Cepeda C1,
Qiao J1, Chang J1, Huynh M1, Levine M1, Wu J1, Pedram K1 1UCLA David
Geffen School of Medicine, Los Angeles, CA and 2Harvard Medical School,
Boston, MA.
Purpose of Study: Most cases of Tuberous Sclerosis Complex (TSC) have
been localized to mutations in either the TSC1 or TSC2 genes. Current
research has yet to establish how different types of mutations (deletions and
polymorphism) affect the variability in neurological disease phenotype. The
aims of this study are to identify associations between types of genetic
mutations and clinical characteristics, neuroimaging, cellular electrophysiology, brain tissue anatomy, and neuronal protein concentrations in TSC
patients undergoing epilepsy neurosurgery.
Methods Used: Data were compiled for twenty pediatric patients (n=20) who
developed epilepsy as a consequence of TSC. Genetic mutations were grouped
into large frameshift mutations or deletions, single nucleotide polymorphisms
(SNPs), and no detected TSC1 or 2 mutation. Clinical characteristics and imaging data were extracted from patient medical records and biochemical techniques, including Western blots and immunohistochemistry, were utilized on
surgical brain tissue to obtain electrophysiological, anatomical, and polypeptide
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
data. The correlation between these variables will be determined through the
application of ANOVA statistics.
Summary of Results: Most mutations have been localized to the TSC2
gene; only 1 has been localized to TSC1. Preliminary data for clinical
characteristics, cellular electrophysiology, anatomical features, protein concentrations, and neuroimaging have been compiled. However, statistical
analysis remains to be conducted, as data about the patients_ developmental
statuses and the EEG characterizations of their seizures must still be obtained.
Due to the single-blinded design of the study, the nature of genetic mutations
will be revealed and accounted for when the remaining information has been
obtained.
Conclusions: We hypothesize that genotype will correlate with phenotype.
More specifically, patients with the largest genetic defects in TSC mutations
will present with more dysmorphic giant cells and balloon cells, increased
concentrations of GABAergic post-synaptic proteins, greater abnormalities in
cellular electrophysiology, and worse post-operative outcomes.
402
ORBITAL DISTANCES DO NOT CORRELATE TO
CEREBELLAR VOLUME IN CHILDREN WITH AUTISM
SPECTRUM DISORDERS
Camilleri K1,2,4, Angkustsiri K2,3, Nordahl CW2, Lee A2, Boyd SB2,3,
Amaral DG2 1UC Davis School of Medicine, Sacramento, CA; 2UC Davis
MIND Institute, Sacramento, CA; 3UC Davis Medical Center, Sacramento,
CA and 4WSMRF, Monterey, CA.
Purpose of Study: Abnormal cerebellar volumes have been reported in
autism spectrum disorders (ASD). One of the hypothesized etiologies of ASD
may be some sort of embryological insult, which could manifest as abnormal
brain development and facial dysmorphisms. Orbital distances might serve as
a simple, external marker for children who have abnormal brain development
since the cerebellum and the orbits develop around the same time (cerebellum: beginning day 32, eyes: days 29-56). We hypothesize that orbital distances are related to cerebellar volume in children with ASD.
Methods Used: 44 Caucasian, male children with ASD between the ages
of 2.2 and 3.7 years old (mean age=3.0, std dev=.42) from the UC Davis
MIND Institute_s Autism Phenome Project were included in this study.
Three-dimensional surface images of the children_s faces were landmarked
using 3dMD software to obtain intercanthal distance, biocular width, and
intercanthal index. Cerebellar volumes were obtained from structural T1
brain MRIs first using a template-based automated segmentation procedure
followed by refinements done in Mayo Clinic BIR_s Analyze 10.0 software.
Linear regression was used to evaluate the relationship between the individual orbital distances and cerebellar volume.
Summary of Results: Cerebellum volume does not significantly correlate
to intercanthal distance (r2= 0.0012, p=0.82), biocular width (r2=0.0024,
p=0.75), or intercanthal index (r2=0.0095, p=0.53).
Conclusions: The lack of any significant correlation between cerebellum
volumes and orbital measurements suggests that the anthropometric measurements may not be good indicators of cerebellum volume in children with
autism. In children with autism that may have suffered an embryological
insult, cerebellum and orbital development may not be similarly affected.
403
MULTIPLE ESTROGEN RECEPTORS ARE RESPONSIBLE
FOR THE ESTROGENIC ATTENUATION OF THE
CANNABINOID-INDUCED INHIBITION OF GLUTAMATE
RELEASE AT PROOPIOMELANOCORTIN SYNAPSES
Washburn N, Wagner E Western University, College of Osteopathic Medicine
of the Pacific, Pomona, CA.
Purpose of Study: Estrogens exert far-reaching effects on mammalian biology - from reproduction to energy homeostasis. While many actions require
gene transcription and protein synthesis to alter cell function via the activation of
estrogen receptor subtypes ER> and ERA (which takes hours to days), it is now
well recognized that estrogenic signaling can take place on a much more rapid
timescale (minutes) via membrane delimited ER> or a Gq-coupled membrane
ER (Gq-mER). We have shown previously that estrogen rapidly diminishes
cannabinoid-induced hyperphagia; attributable to an attenuation in the
cannabinoid-induced presynaptic inhibition of excitatory glutamate currents
at anorexigenic POMC neurons of the hypothalamic arcuate nucleus. Recently,
we found that activation of ER> and Gq-mER attenuated cannabinoid-induced
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
hyperphagia, whereas ERA did not. Therefore it was the goal of the present
study to further investigate whether ER> or Gq-mER, contribute significantly to
the rapid and sustained inhibitory effect of estrogen on the cannabinoid regulation of POMC neuronal activity.
Methods Used: Voltage clamp experiments were performed in the arcuate
nucleus of female Topeka guinea pigs. Once stable access to the cell was
achieved either the ER> agonist PPT (1KM), the Gq-mER agonist STX
(10nM), or their respective vehicle controls, were bath applied for 10-15 min.
Miniature excitatory postsynaptic current (mEPSC) frequency and amplitude
were then evaluated under both basal conditions and in the presence of the
cannabinoid receptor agonist WIN 55,212-2.
Summary of Results: Transient exposure to STX diminished the decrease
in mEPSC frequency caused by 1 KM WIN 55,212-2, whereas PPT did not
(vehicle: 49.6T9.1%; STX: 85.8T4.2%; PPT: 45.0T4.8% of baseline control).
However, systemic administration of PPT (200KM; s.c.) completely blocked
the cannabinoid-induced decrease in mEPSC frequency (105.8T4.5% of
baseline control).
Conclusions: In conclusion, Gq-mER and ER> contribute to different
components of estrogenic attenuation of the cannabinoid-induced decrease in
glutamate release at POMC synapses: Gq-mER mediates the rapid inhibition,
whereas ER> is involved in the sustained diminution.
404
MTOR SIGNALING IN ANGELMAN SYNDRONE
Hsu R, Moreno S, Liao G, Bi X College of Osteopathic Medicine, Western
University of Health Sciences, Pomona, CA.
Purpose of Study: Angelman syndrome (AS) is caused by the loss of the
normal maternal contribution to a region of chromosome 15. In a normal
individual, the maternal allele is expressed and the paternal allele is silenced.
If the maternal contribution is lost or mutated, the result is AS. It should be
noted that the methylation test that is performed for AS results in a defect in
UBE3A causing human neurological disorder. Other neurodegenerative
disease research such as Alzheimer_s and Huntington_s disease have found
that there are dysregulation in the mammalian target of rapamycin (mTOR)
pathway, which play a crucial role in neuronal development and synaptic
plasticity. However, it remains unclear whether mTOR signaling is disrupted
in AS.
Methods Used: In our study, AS mice models (Ube3a maternal-deficient
mice) and wildtype (Ube3a) were used to compare different mTOR cellular
expression in mice tissue. Immunohistochemical staining followed by confocal
microscopy was used to examine the expression of p-mTOR-2448, p-mTOR2481, LAMP-1, p70S6K, Beclin-1, PS-1 in the motor cortex, somatosensory
cortex, hippocampus, and cerebellum. The images were analyzed using ImageJ
program calculating the number of protein involvement and the intensity of the
fluorescent.
Summary of Results: Our study showed an increase in mTOR phosphorylated at residues Ser2448 and Ser2481 in the Purkinje cells of the
cerebellum while the motor cortex, somatosensory cortex, hippocampus
showed no significant difference. Similarly, levels of LAMP1 expression, a
lysosome protein, was also increased in Purkinje cells while the motor cortex,
somatosensory cortex, hippocampus also showed no significant difference.
P70S6K, Beclin-1, PS-1 remains unchanged.
Conclusions: Autophagy is highly regulated by mTOR in normal signaling.
The high levels of mTOR should normally inhibit the expression level of
autophagic activity, however, our studies demonstrates that AS mice shows
abnormally high levels of mTOR and autophagic activity especially in LAMP-1,
which implies that there is a lack regulatory control in the pathway. Interestingly,
confocal images showed that mTOR, especially its phosphorylated isoforms,
were mainly localized in neurons in endosome/lysosome-like structures suggesting that AS could also be a lysosomal disease.
405
ASTROCYTE MATURATION AND REACTIVITY IN
PEDIATRIC EPILEPSY SYNDROMES
Kwon EE1,2, Mathern G3, Wanner I2 1Western University of Health Sciences,
Pomona, CA; 2University of California, Los Angeles, Los Angeles, CA and
3
University of California, Los Angeles, Los Angeles, CA.
Purpose of Study: Astrocytes are directly involved in the blood brain
barrier and influence potassium buffering through aquaporin4 (AQP4) water
channels. AQP4 is known to buffer metabolites during neural injury and
201
Western Regional Meeting Abstracts
Journal of Investigative Medicine
epileptogenesis. GFAP (glial fibrillary acidic protein), a prototypical marker
of astrocytes, is upregulated and reveal astrocytes undergo structural changes
such as hypertrophy and loss of domain organization. Severe forms of
astrogliosis lead to the formation of glial scars and scar-like penumbras
around necrotic or ischemic areas. To further characterize astrocytes in
human epilepsy, we analyzed hallmark markers of astrocytes in pediatric
surgical biopsy specimens of the neocortex.
Methods Used: Cortical tissue were resected from pediatric patients with
epilepsy disorders including Cortical Dysplasia, Rasmussen_s Encephalitis,
and epilepsy due to ischemic stroke primarily from central operculum and
parieto-temporal lobe regions. Specimens were vibratome-sectioned and
stained for GFAP and AQP4 using flourescence immunohistochemistry and
images were binarized and quantified for astrocyte abundance and coverage
as a measure of reactivity.
Summary of Results: Our results show that abundance as well as network
complexity varied in each zone. One striking finding was a noticeable
sparseness, even lack GFAP fluorescence in the grey matter of infant cortical
biopsies. This finding, together with the presence of immature radial glia
marker expressing cells in that zone (BLBP-brain lipid binding protein)
suggests that GFAP-positive astrocytes did not significantly populate the grey
matter until around 4 years of age. This was found to be the case in genetic as
well as acquired epilepsy etiologies, irrespective of case severity, which
suggests a developmental feature of protoplasmic astrocytes rather than
pathological.
Conclusions: These tools allow us to determine reproducible quantitative
differences in astrocyte abundance, morphology and reactivity that may help
define gliosis severity. These findings would help us understand the role of
astrocytes in epilepsy and also be useful in histopathological diagnosis of
various epilepsy syndromes.
Pulmonary and Critical Care II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
406
ACCELERATED PPARF SIGNALING IN POST
PNEUMONECTOMY LUNG REPAIR
Li Y1, Sakurai R1, Vonswinckel R2, Torday J1, Rehan V1 1Los Angeles
Biomedical Institute, Torrance, CA and 2University of Giessen, Giessen,
Germany.
Purpose of Study: Peroxisome proliferator-activated receptor (PPAR)F, a
key nuclear transcription factor, plays an essential role in lung development
and injury/repair in the newborn, but its role in pulmonary regeneration in the
adult is not known. Here, we aimed to determine the role of PPARF signaling
in adult lung injury/repair, as a prelude to manipulate molecular determinants
for the purpose of enhancing lung regeneration in the adult.
Methods Used: We utilized a post-pneumonectomy (post-PNX) compensatory lung growth model. Adult C57BL/6 mice underwent left-sided PNX.
The right lung was examined on days 2, 5, and 14 post-PNX. Lung volume
was assessed, and PPARF signaling pathway intermediates were examined at
the RNA (real time RT-PCR) and protein levels (immunohistochemistry and
Western blot analysis).
Summary of Results: Following PNX, right-lung volume rapidly compensated for the initial volumes of the left and right lungs. Both Western blot
analysis and real time RT-PCR suggest that compared to sham controls,
following PNX, PPARF was up-regulated (+31% from post-PNX day 2 to
day 5) and this increase was normalized by day 14 post-PNX. Qualitatively, a
similar pattern was demonstrated by down-stream targets of PPARF signaling
such as adipocyte differentiation-related protein (ADRP) and leptin, though
the changes were more pronounced for ADRP. These data were corroborated
by immunohistochemistry.
Conclusions: Our data suggest clear up-regulation of PPARF signaling
during post-PNX lung repair, suggesting a role for this critical pathway in
adult lung injury/repair. We speculate that PPARF agonists could also accelerate adult lung injury repair, similar to what we have recently shown in
neonates (Dasgupta et al, Am J Physiol 2009;296:L1031-41). (Grant support:
NIH-HL55268, HL075405, HD058948, HD051857) and TRDRP (15IT0250, 17RT-0170).
202
&
Volume 59, Number 1, January 2011
407
IMPACT OF VITAMIN D DEFICIENCY ON OUTCOME IN
CRITICALLY ILL PATIENTS
Higgins D1, Queensland K1, Wischmeyer P1, Sufit A1, Heyland D2
1
University of Colorado School of Medicine, Denver, CO and 2Queen’s
University, Kingston, ON, Canada.
Purpose of Study: Although as many as 1 billion people worldwide have a
deficiency in vitamin D which has been associated with a myriad of disease
conditions, the impact of this deficiency on outcome in critically ill patients is
not clear. We hypothesize that vitamin D deficiency in ICU patients is a
common condition and is associated with negative outcomes.
Methods Used: We conducted a prospective multi-center study of baseline
serum vitamin D levels in 195 patients admitted to the ICU to evaluate the
incidence of vitamin D deficiency and associations with mortality, infection
status, length of stay, and organ function. Infection status and clinical outcomes were documented for 28 days after admission. A Cox proportional
hazard model, logistic regression and ANOVA models with adjustment for
age, BMI and APACHE II score were used.
Summary of Results: Vitamin D deficiency or insufficiency (G60 nmol/L)
was observed in 159 patients (82%). Vitamin D status was not associated with
28-day all cause mortality (hazard ratio (HR) 1.004; 95% CI 0.993Y1.015).
However, increased levels of vitamin D were associated with a shorter length
of stay (HR 3.56; 95% CI 1.5Y8.6 at 28 days). Vitamin D deficient and
insufficient patients had a higher incidence of probable infections compared
to patients with normal levels of vitamin D (OR 3.2; 95% CI 0.78Y13.1,
p=0.1). Patients with culture confirmed pneumonia had significantly lower
levels of baseline vitamin D (38.6 nmol/L +/j 3.2) compared to those who
did not have pneumonia (48.23 nmol/L +/j 1.9) (pG.01). Lower levels of
baseline vitamin D also correlated with increased baseline SOFA scores
(pG.05), and elevated serum Cr levels (pG.01).
Conclusions: This is largest prospective cohort trial to date showing vitamin
D insufficiency is a common condition in the ICU and the first data showing a
significant correlation of Vitamin D deficiency with longer length of ICU
stay, increased risk of pneumonia, and elevated organ failure scores. This may
be due the vital role of vitamin D in regulating immune function.
408
CLINICAL UTILITY OF EXHALED NITRIC OXIDE
ASSAY IN ASSESSMENT OF ASTHMA CONTROL
IN CHILDREN
Vijayvargiya P, Joseph M UCSF-Fresno, Fresno, CA.
Purpose of Study: Clinical assessment of asthma control is done primarily
using symptom review of asthma impairments and risk factors (2007 NIH
Asthma Management Guidelines). The only objective tool for asthma control
assessment is the estimation of predicted % of FEV1 and FEV1/FVC. Assay
of exhaled Nitric Oxide (eNO) is a relatively new and non-invasive test
considered useful in asthma control assessment. The purpose of this pilot
study is to compare the accuracy of eNO, FEV1 and FEV1/FVC as objective
tools for asthma control assessment. The study was done in an urban, pediatric asthma clinic in Fresno, CA.
Methods Used: A retrospective chart review of children with persistent
asthma 5-18 years of age, on inhaled corticosteroids attending asthma clinic
during the past 2 years was done. Asthma control was classified as either
Fwell controlled_ or Fnot controlled_. FEV1 and FEV1/FVC were estimated
by office spirometry performed by the same trained technician in all cases.
Only spirometry curves that met the 2005 ATS acceptability criteria were
included. Level of asthma control based on symptom review, corresponding
FEV1, FEV1/ FVC and eNO levels on the same visit were obtained. The
accuracy of the three continuous test variables was analyzed using a ROC
analysis. Area under the curve and confidence interval were calculated using
non-parametric methods.
Summary of Results: We analyzed 92 data sets from 62 asthmatics,37
males and 25 females. Asthma was documented as Fwell controlled_ in 41 and
Fnot controlled_ in 51 visits. Among the three test result variables, both FEV1
and eNO were found to be useful, but eNO appeared to be a better tool for
assessing asthma control. FEV1/ FVC ratio was not found to be useful. At the
established cut off level of 20ppb of eNO as the upper limit of normal in
children, the specificity for predicting asthma control was 82% and sensitivity
55% in this cohort. The area under curve for ROC analysis for eNO was
0.759 (95% CI, 0.66Y0.85; pG0.001).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Western Regional Meeting Abstracts
Conclusions: Estimation of eNO proved to be a better objective tool for
Summary of Results: Out of the 23 questions, 12 questions were intended
assessing asthma control than FEV1 estimation in our cohort from this pilot
study. Our results indicate that eNO assay is an alternate, non-invasive objective tool to assess asthma control. It is a useful aid in clinical decision
making process especially when history is unreliable and spirometry is either
difficult or unavailable.
to test the resident_s theoretical knowledge, 8 questions to test their ability to
assess U/S images, and 8 questions to assess their U/S practical knowledge.
Their scores were as follows: 42.25% of the interns answered the questions
correctly on the theoretical knowledge section vs. 26.3% on assessing U/S
images, and 29.3% on the practical U/S knowledge section. The difference
between theoretical knowledge was statistically significant (p=0.05), however
the difference between theorical and practical knowledge was only a trend.
Conclusions: Interns beginning their clinical rotations have greater background knowledge in theoretical concepts of U/S guided central line placement than ability to interpret U/S images. Curriculae designed to teach this
skill to beginning interns should place special emphasis on the development
of image interpretation skills. Our presentation will also include 3-month
post test data (currently being collected) to evaluate their improvement with
instruction and their late retention.
409
GENETIC POLYMORPHISMS OF CYTOCHROMES P450
AND PEDIATRIC ASTHMA CONTROL BY INHALED
CORTICOSTEROIDS
Stockmann C1, Fassl B1, Nkoy F1, Stone B1, Willis L1, Gaedigk R2, Leeder S2,
Ward R1 1University of Utah, Salt Lake City, UT and 2University of Kansas
Children’s Hopaital, Kansas City, KS.
Purpose of Study: Inhaled corticosteroids (ICS) are mainline treatments for
persistent asthma in children. Up to 10% of children with persistent asthma
fail ICS therapy for unclear reasons. Genetic polymorphisms of cytochromes
P4503A (CYP3A) have been shown to be associated with altered metabolism
of ICS which might have an effect on their therapeutic effectiveness.
Fluticasone, a widely used ICS, is metabolized primarily by CYP3A4 whose
metabolism may be decreased by a polymorphism in intron 6. Objectives of
this study are to 1) determine the frequency of genetic variants in CYP3A4
(intron 6 SNP rs35599367,C9T), in children treated with fluticasone for
difficult to control asthma and 2) to describe and compare CYP450 genotypes
in these children with and without previous hospital admissions for asthma.
Methods Used: We enrolled children aged 2Y17 years with a diagnosis of
asthma from Primary Children_s Medical Center. We collected saliva samples
and analyzed for CYP450 genetic polymorphisms at the Developmental
Pharmacology and Experimental Therapeutics Laboratory at Kansas City.
Single nucleotide polymorphisms for 9 alleles that increase or decrease
protein expression and CYP3A activity were determined along with asthma
severity, preventive medication use, and number of hospitalizations for acute
asthma within the preceding 12 months.
Summary of Results: Of 96 children enrolled, 23 were treated with fluticasone; all of these were classified as not-well-controlled asthmatics by NIH
guidelines. Of these 23 children, 14 patients were admitted for asthma in the
previous 12 months, 14/14 featured an rs35599367 C/C genotype consistent
with greater CYP3A4 mRNA level and enzyme activity. Of the 9 patients not
requiring hospitalization within the last 12 months, 5 (55.6%) had at least one
variant rs35599367 allele, consistent with slower metabolism of fluticasone.
There was no difference in the distribution of other CYP3A4, CYP3A5, and
CYP3A7 polymorphisms.
Conclusions: These preliminary findings are consistent with the hypothesis
that slower metabolism of ICS may improve their effectiveness for treatment
of asthma. Additional patient studies are needed to confirm or refute this
association.
410
INTERN NEEDS IN ULTRASOUND GUIDED CENTRAL
VENOUS ACCESS DIDACTIC TEACHING
Abouhouli H, Boivin M, Baty G UNM, Albuquerque, NM.
Purpose of Study: Residency Training involves teaching critical procedural skills, including invasive and sometimes dangerous procedures. Central
venous catheter (CVC) placement is a core procedural skill that typifies this
subset. Our group sought to develop a curriculum and test different strategies
to teach this multifaceted skill. There is no current standardized approach to
training residents in CVC placement. Our project is designed to determine the
needs within didactic training that would include theoretical and practical
aspects of U/S guided CVC placement aimed toward the interns prior to their
ICU rotation.
Methods Used: We have randomly selected 45 interns to participate in the
above study. Out of the 45 residents 23 completed the pre-test. The questions
were designed to test the theoretical knowledge, ability to assess U/S images,
and the U/S general knowledge of interns at the start of their year. The
questions were validated in ultrasound skilled groups prior to administration.
Some of the questions had actual U/S images and the interns were asked to
properly identify the marked structures. The pre-test is an online computer
based test, each participant had access to the website, and they were allowed
to take the test off campus, at their own pace.
* 2011 The American Federation for Medical Research
411
INHIBITION OF FARNESYLTRANSFERASE WITH FTI-277
EXACERBATES EOSINOPHILIC INFLUX IN ALLERGIC
AIRWAY INFLAMMATION
Zeki AA1,2, Chang KY2, Kenyon N1,2 1U.C. Davis Medical Center, Sacramento,
CA and 2Center for Comparative Respiratory Biology & Medicine (CCRBM),
Davis, CA.
Purpose of Study: We have previously shown that simvastatin inhibits
allergic eosinophilic inflammation and improves lung function in the ovalbumin mouse model by targeting the mevalonate (MA) pathway. Farnesyltransferase (FTase) is a downstream enzyme in the MA pathway that activates
Ras, a small GTPase important in Th2-dependent eosinophilic inflammation.
Thus, we hypothesized that inhibition of FTase with the drug FTI-277 would
attenuate allergic airway inflammation and improve lung function.
Methods Used: BALB/c mice were sensitized to ovalbumin (OVA) over
4 weeks, then exposed to 1% OVA aerosol over 2 weeks. Mice were injected
with FTI-277 (20 mg/kg/day for 14 days) intraperitoneally before each OVA
exposure. Bronchoalveolar lavage fluid (BALF) total and differential cell
counts, lung histology, and lung compliance and resistance were measured
(using a plethysmograph for restrained animals).
Summary of Results: Control and treated animals showed no significant
change in body weight. FTI-277 markedly increased total cell, eosinophil
(pG0.05), and lymphocyte counts (pG0.05) in BALF. Lung histology showed
marked peribronchiolar and perivascular influx of eosinophils, lymphocytes,
and neutrophils in FTI-277 treated mice. Systemic FTI-277 also increased
airway hyperreactivity (AHR) (pG0.01) and decreased lung compliance
(pG0.05).
Conclusions: Systemic treatment with FTI-277 exacerbated allergic eosinophilic inflammation in a mouse model of asthma. This is thought to occur
via inhibition of Ras activity. These data suggest that inhibition of certain
downstream targets in the MA pathway may be harmful, while others may
have benefit.
Supported by the following grants: NIH (T32) HL07013, NCRR UL1
RR024146 (K30), HL-076415 (K08), ATS Fellows Career Development
Award, and CTSC K12 Award (KL2 RR 024144).
412
HUMAN PULMONARY ARTERY ENDOTHELIAL CELL
EXPOSURE TO FIBRIN(OGEN) AUGMENTS
INTRACELLULAR CALCIUM RESPONSES TO THROMBIN
Liang N, Firth A, Marsh J, Yuan J, Morris T University of California
San Diego, San Diego, CA.
Purpose of Study: After pulmonary embolism, some patients are at risk for
developing chronic thromboembolic pulmonary hypertension (CTEPH). Fibrin from patients with CTEPH is resistant to lysis, which leads to persistence
within thrombi of fibrin(ogen), although how this phenomenon might stimulate pulmonary arterial scar formation is unknown. This study was performed to determine if exposure of human pulmonary artery endothelial cells
(PAEC) to regions of fibrin(ogen) would enhance their ability to be stimulated
by subsequent exposure to thrombin.
Methods Used: The central BN-terminus disulfide knot[ (NDSK), was
prepared from human fibrinogen by digestion with cyanogen bromide. NDSK
II (the central region of fibrin) was prepared by cleavage of NDSK with
thrombin. Ca2+ was labeled using Fura-2. PAECs were exposed to NDSK,
203
Western Regional Meeting Abstracts
Journal of Investigative Medicine
NDSK II or control then subsequently stimulated with thrombin. Thrombininduced [Ca2+]cyt increases (i.e. Fura-2 fluorescence) were measured in
peripheral cytoplasmic regions from cells by epifluorescence microscopy.
Data were collected at a rate of 88 samples/minute using excitation 340 and
380 nm with xenon lamp, emission 520 nm at 20 magnification.
Summary of Results: Peak thrombin-induced increases in [Ca2+]cyt
(F340/F360) after pre-perfusion with NDSKII (0.914 +/j 0.139) was significantly higher than after pre-perfusion with control (0.537+/j 0.028;
p G 0.001). The AUC for thrombin-induced [Ca2+]cyt increase was also
higher after NDSKII (73.56+/j 14.02) than after the control (32.49+/j 2.49;
pG 0.0001). There was a trend towards higher thrombin-induced [Ca2+]cyt
increases in cells pre-perfused with NDSK (0.808+/j0.049) compared with
control (0.716+/j 0.051, p=0.2). The AUC for thrombin-induced [Ca2+]cyt
increase was significantly higher after NDSK (78.19+/j 6.97) than after
control (35.89+/j 4.25; pG 0.0001).
Conclusions: Exposure of endothelial cells to the cell signaling regions of
fibrin and, to a lesser extent, fibrinogen enhanced their ability to be activated
by thrombin. The enhancement may mediate the fibrin-endothelial cell
interactions responsible for pulmonary embolism remodeling and persistent
vascular obstruction in CTEPH.
413
UNDIAGNOSED AIRFLOW OBSTRUCTION, OBESITY
AND RESPIRATORY SYMPTOMS: A COMPLEX
INTER-RELATIONSHIP
Zutler M, Singer J, Omachi TA, Blanc PD University of California, San
Francisco, San Francisco, CA.
Purpose of Study: Screening spirometry detects previously undiagnosed
airflow obstruction (AO) in up to 15% of the general adult population. Although increased cough and dyspnea are associated with AO, other covariates, in particular body habitus, have not been well characterized. We sought
to explore the inter-relationships among AO, obesity, and self-reported dyspnea on exertion (DOE).
Methods Used: We analyzed the referent group of Kaiser Permanente
Health Plan members (age 40Y65) recruited for the Function, Living, Outcomes and Work (FLOW) study of chronic obstructive pulmonary disease
(COPD). The referent group had no health utilization for COPD in the 12
months prior to recruitment. Subjects completed a structured interview
(medical history, symptoms and functional status) and a clinical assessment
including body mass index (BMI) and spirometry. We excluded 2 subjects
with AO (FEV1/FVC ratio G0.7) who self-reported a history of physiciandiagnosed COPD. We compared smoking and obesity among subjects with
AO to those without AO. In multiple regression analyses (controlling for age,
sex, race-ethnicity and cumulative smoking), we tested the associations of
AO, obesity (BMIQ30), and dyspnea on exertion.
Summary of Results: Of the 371 subjects studied, 69 (18.6%, 95% CI 14.622.6%) manifested AO. Among those with AO, 43% were never-smokers
compared to 52% among those without AO (pG0.01). Among former and current smokers, those with AO had a higher cumulative smoking history than those
without AO (median 27 vs. 19 pack-years, p=0.02). In multivariate analysis,
obesity was associated with an increased likelihood of DOE (OR 4.0; 95%CI
2.0Y7.8) as was female gender (OR 2.0; 95%CI 1.1Y3.7). AO trended towards an
increased likelihood of DOE (OR 1.7; 95%CI 0.7Y4.6). Obesity was negatively
associated with AO (OR 0.54; 95%CI 0.3Y0.98).
Conclusions: AO is common in adults without an established COPD diagnosis. Cumulative smoking was associated with AO, but, counter to
expectations, obesity was negatively associated with AO. Both female gender
and obesity were linked to a higher likelihood of DOE; AO trended in the
same direction. These data suggest that respiratory symptoms are related to
complex inter-relationships among body habitus, gender, and lung function.
414
SURVIVAL IN SCLERODERMA-RELATED INTERSTITIAL
LUNG DISEASE FOLLOWING LUNG TRANSPLANT
Sottile PD1, Iturbe DF3, Katsumoto T1, Connolly KM1, Collard H1, Golden JA1,
Hoopes C2, Singer JP1 1UCSF, San Francisco, CA; 2UCSF, San Francisco, CA
and 3Hospital Universitario Marques de Valdecilla, Santander, Spain.
Purpose of Study: Interstitial lung disease (ILD) is the leading cause of
death in scleroderma. A diagnosis of scleroderma-related ILD (Scl-ILD) is
204
&
Volume 59, Number 1, January 2011
often a contraindication for lung transplant due to concern that the extrapulmonary organ involvement of scleroderma will result in poorer outcomes.
In this study, survival following lung transplant in patients with Scl-ILD was
compared to idiopathic pulmonary fibrosis (IPF), and idiopathic pulmonary
hypertension (IPH).
Methods Used: We conducted a cohort study from 2001 to 2010. Patients
transplanted for Scl-ILD were matched by age and gender to control patients
transplanted for IPF; patients transplanted for IPH were also included as
controls. Esophageal dysfunction was not a contraindication for transplant.
Demographic and physiologic data were collected. Post-transplant survival
time was the primary outcome. Survival time was estimated using KaplanMeier methods. Secondary outcomes included episodes of acute rejection
and time to development of bronchiolitis obliterans syndrome.
Summary of Results: Twenty-two patients with Scl-ILD, 30 patients with
pulmonary fibrosis, and 11 patients with IPH who underwent lung transplant
were identified. Scl-ILD and IPF patients were similar in age, gender, ethnicity, body mass index, serum creatinine, FEV1, and FVC. Most patients
with Scl-ILD had esophageal dysmotility and/or gastroesophageal reflux.
Patients transplanted for Scl-ILD had similar overall survival to patients
transplanted for IPF and IPH (p=0.4). In the Scl-ILD group, 6, 12 and
36 month survival was 85% 79%, and 79% compared to 90%, 87%, and 69%
in the IPF group. In IPH, 6, 12 and 36 month survival was 90%.
Conclusions: Patients transplanted for Scl-ILD appear to have similar survival following lung transplantation compared to patients transplanted for IPF
and IPH. For patients with end-stage Scl-ILD, referral to institutions with
experience in performing transplant for this disease should be considered.
Future studies are needed to determine whether these similarities in survival
persist, whether there are important differenced in morbidity, and whether
center experience and volume are important factors in Scl-ILD patient
outcomes.
415
THE EFFECT OF INFLAMMATION AND MECHANICAL
VENTILATION ON ALVEOLAR FLUID CLEARANCE
IN MICE
Loraas EK1,2, Smith L1,2,3, Martin TR1,2 1University of Washington, Seattle,
WA; 2VA Puget Sound Medical Center, Seattle, WA and 3Seattle Children’s
Hospital, Seattle, WA.
Purpose of Study: Acute lung injury (ALI) is a syndrome of acute
hypoxemic respiratory failure that affects more than 190,000 people each
year in the United States, and results in over 74,000 deaths annually.
Infections are the most common cause of ALI, and animal studies suggest that when infectious stimuli are combined with mechanical ventilation (MV) there are synergistic increases in the inflammatory and
injury responses in the lungs. The accumulation of a protein rich alveolar
edema fluid is a pathologic hallmark of ALI, and resolution requires an
intact alveolar epithelium to clear the fluid. Impaired alveolar fluid
clearance (AFC) is associated with mortality in adult patients with ALI.
Therefore, we investigated the effect of inflammation and mechanical
ventilation on AFC in adult mice.
Methods Used: Adult (12 week) C57BL/6 mice were treated with
0.36 ng/g intratracheal lipopolysaccharide (LPS) followed by mechanical
ventilation using Vt=15 ml/kg and rate=80 breaths/min for four hours
(LPS+MV). Controls were untreated age-matched, spontaneously breathing mice. Animals were euthanized and fluorescent labeled albumin was
instilled into the airways through a tracheostomy tube. Animals were then
placed on 5 cm H2O continuous positive air pressure and 100% O2 for
30 minutes. The fluorescent labeled albumin was aspirated from the lungs
and measured using fluorescent absorption spectroscopy. The difference
between the initial albumin fluorescence and recovered albumin fluorescence was used to calculate AFC. Cytology was also performed on the
recovered fluid.
Summary of Results: Treatment with LPS+MV (N=8) resulted in a significant recruitment of neutrophils (75% PMN) into the lungs as compared
with untreated control mice (N=10, 0% PMN). Treatment with LPS+MV
resulted in a mean AFC of 48.7 T 5.1, whereas the mean AFC of untreated
control mice was 48.2 T 2.3.
Conclusions: The combination of LPS and mechanical ventilation causes a
significant neutrophil recruitment into the lungs. Surprisingly, the inflammatory response to LPS+MV does not change AFC, suggesting that the
alveolar epithelium remains functionally intact.
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
416
METABOLOMIC ANALYSIS OF EXHALED BREATH
CONDENSATES (EBC) IN ASTHMA AND COPD
Schivo M1, Zhao W2, Aksenov A2, Kenyon N1, Davis C2 1UC Davis,
Sacramento, CA and 2UC Davis, Davs, CA.
Purpose of Study: A significant proportion of patients do not fit classic
historical or spirometric definitions of either asthma or COPD and therefore
elude a confident diagnosis. Metabolomic profiling may provide a better
diagnostic strategy and may eventually elucidate specific therapeutic targets
in asthma and COPD. A novel device called a differential mobility spectrometer (DMS) is a portable, highly sensitive sensor. In this study we aim to
test EBC from asthma, COPD, and health subjects using a DMS. We hypothesize that EBC chemical profiles will differ between subject groups.
Methods Used: 20 subjects were assigned to an asthma, COPD, and nonasthmatic/bronchitic control group based on well-characterized clinical and
spirometric criteria. Both nitric oxide and exhaled breath condensate were
collected as well as a St. George_s Respiratory Questionnaire and anthropomorphic measures. The nitric oxide was measured on an analyzer, and the
exhaled breath condensate was introduced into a gas chromatogram and
analyzed by both a differential mobility spectrometer (DMS) and a mass
spectrometer (MS). Optimization of EBC volatile compound recovery involved modifying heating time, sample agitation, sample volume, vial size,
and background noise reduction. DMS and mass spectrometer results were
compared to note if similar compounds exist, some of which can only be
identified in the mass spectrometry library.
Summary of Results: Subject groups differed by age; subjects with COPD
were significantly older than either those with asthma or the controls
(PG0.05). DMS data is now being analyzed for peak-to-peak differences
(t-testing), biomarker pattern differences (principle component analysis), and
individual compound identification between groups. We anticipate that
principle component analysis of the differential mobility spectrometer data
will allow for accurate separation of the three groups. We also anticipate that
biomarker Bsignatures[ will emerge unique to each group from the DMS and
MS data.
Conclusions: The DMS can measure large profiles of metabolites in exhaled
breath condensate of subjects with asthma and COPD. PCA analyses will
allow us to determine which metabolites distinguish asthma from COPD, and
we plan to present these determinations. Age was A confounder in our study.
Surgery II
Concurrent Session
8:30 AM
Saturday, January 29, 2011
417
SIROLIMUS DETERS THE RATE OF HEPATITIS C
PROGRESSION FOLLOWING ORTHOTOPIC LIVER
TRANSPLANTATION: A SINGLE CENTER EXPERIENCE
Kelly MA, Kaplan M, Zimmerman MA, Wachs M, Bak T, Kam I University
of Colorado Health Sciences Center, Aurora, CO.
Purpose of Study: Recurrence of hepatitis c virus (HCV) infection is a
foreseeable problem following orthotopic liver transplantation (OLT), which
over time leads to liver fibrosis and graft loss. While several animal studies
have shown that sirolimus (SRL) acts to inhibit the rate of liver fibrosis, few
studies in the human population support or refute these findings. Our center
has had ample experience with using SRL as an effective immunosuppressive
agent. In this study we sought to determine the difference in the rate of
recurrence of HCV infection as well as the progression of infection in patients
who received SRL as primary immunosuppression in comparison to patients
who received calcinurin inhibitors (CNIs) post-OLT.
Methods Used: Patients transplanted for end-stage liver disease (ESLD)
due to HCV were identified from our transplant database. These patients were
categorized into two groups: CNI immunosuppression (control group) and
SRL immunosuppression group. We controlled for 3 clincopathologic variables (MELD, warm ischemic time, and cold ischemic time). Cox proportional hazards regression was used to examine the effect of Sirolimus on
overall mortality as well as severe HCV recurrence (defined as a liver biopsy
of stage 2 fibrosis or greater). Logistic regression analysis was used to test the
association between Sirolimus and severe HCV recurrence within in the first
year following OLT.
* 2011 The American Federation for Medical Research
Western Regional Meeting Abstracts
Summary of Results: From January 2000 to July 2009, 313 patients underwent OLT for HCV. After applying exclusion criteria our study population
was 232 patients, of which 60 patients received SRL as primary immunosuppression therapy. Overall, no effect was detected of SRL on mortality. For
severe HCV recurrence within the study period, SRL had a statistically significant reduction in risk of the outcome by 58% (p=0.011) and a trend toward
significant reduction in the odds for severe HCV recurrence within the first
year following OLT (p=0.072).
Conclusions: While a debate exists about the effect of SRL therapy on
recurrence of HCV infection following OLT, we demonstrated a significant
reduction in risk in this single-center experience. Furthermore, SRL therapy
showed a trend toward significance in reducing the odds of severe HCV
recurrence within the first year following OLT.
418
PRE-LIVER TRANSPLANT THERAPIES FOR
HEPATOCELLULAR CARCINOMA CAN ACHIEVE
EQUIVALENT SURVIVAL FOR PATIENTS WITH WORSE
PROGNOSIS
Klingler K, Mangus S, Maluccio M Indiana University School of Medicine,
Indianapolis, IN.
Purpose of Study: Liver transplantation is the most effective treatment for
patients with cirrhosis and hepatocellular carcinoma (HCC). Patients often
undergo liver-directed therapies (LDT) aimed at controlling tumor growth
while they await transplantation, or to downsize tumors to meet listing criteria
for transplantation. The study examines the impact of LDT on post-transplant
survival.
Methods Used: Records from a prospectively collected database of 1116
consecutive adult liver transplants performed over a 9-year period were
reviewed. Of these patients, 244 (22%) were diagnosed with HCC by explant
pathology, with median follow-up time of 61 months. LDTs included any
localized intervention made to decrease or slow the progression of the HCC.
Summary of Results: There were 124 patients (51%) who received, and
120 patients (49%) who did not receive any form of LDT. Those who received LDT had significantly larger (median 3.3cm vs 2.0cm, p G0.001) and
more numerous (mean 2.5 vs 1.9, p=0.001) tumors, and were more likely to
be outside Milan criteria (71% vs 29%, p G0.001) and to have neurovascular
invasion on explant pathology (22% vs 14%, p=0.12). In spite of the LDT
group having less favorable tumor characteristics, there was similar survival
(p=0.51) over a 5-year follow up period by Cox proportional hazards regression modeling.
Conclusions: Patients with HCC who undergo LDT to slow progression or
to downsize tumors prior to transplantation can achieve survival similar to
that seen in patients with a much more favorable tumor status and similar to
that seen for all liver transplant recipients at 5-years.
Cox proportional hazards patient post-liver transplant for 244
patients with a history of cirrhosis and hepatocellular carcinoma.
205
Western Regional Meeting Abstracts
Journal of Investigative Medicine
419
IN VITRO MICRODISTRACTION OF HUMAN
ADIPOSE-DERIVED STEM CELLS MAY PROMOTE
THEIR OSTEOGENIC POTENTIAL
Lee JC, Fan K, Sorice S, Bradley J, Zuk P David Geffen School of Medicine
at UCLA, Los Angeles, CA.
Purpose of Study: Distraction osteogenesis has been used over the past 15
years to induce bone formation through the application of gradual, outward
linear force. The molecular mechanisms in which these forces promote bone
formation in the field of craniofacial reconstruction are not well understood.
In an effort to study the response of osteoblasts to compressive and distractive
forces in a 3D environment, we have developed an in vitro model utilizing a
microdistraction (MD) system capable of subjecting cells to distractive and
compressive forces. Using the MD system, our aim is to characterize the
effect of distraction forces on the expression of osteogenic, cytoskeletal, and
cell cycle-related genes in adipose-derived stem cells (ASCs).
Methods Used: Adipose-derived stem cells were isolated from raw human
lipoaspirates, cultured, and suspended in a type-1 collagen gel fitted to the
MD system. ASC-collagen 1 gels were subjected to 1) outward linear distraction forces (distraction gels), 2) static forces (isometric controls), and 3)
no forces (free floating). At designated time intervals, ASCs were harvested
and gene expression for markers of osteogenesis, cell cycle progression, and
cytoskeletal changes were measured using RT-PCR.
Summary of Results: Distraction over four days produced fluctuating
temporal expression patterns in cytoskeletal and cell-cycle markers. While
short-term distraction over four days demonstrated inhibition of osteogenic
Cbfa-1, AP, OP, and OC expression, long-term distraction up to two weeks
resulted in increased levels of Cbfa-1, AP, and ON expression compared to
isometric and free-floating controls.
Conclusions: The application of linear forces on ASCs has complex temporal effects on many markers associated with cell cycle progression and
cytoskeletal changes. Linear stress is capable of promoting the expression of
several key osteogenic genes in our in vitro model. The osteogenic response
to these forces appears to be complex, dependent on the type and timing of
linear stress. Further study is focused on more precisely examining the response of ASCs to distractive manipulation and the roles of the various
intracellular signaling pathways involved.
420
NOVEL RISK FACTORS FOR ANTERIOR CRUCIATE
LIGAMENT INJURY: SHORT LATERAL TIBIAL PLATEAU
LENGTH AND DECREASED LATERAL TIBIAL PLATEAU
RADIUS OF CURVATURE
Westermann RW, Wahl C, Blaisdell G, Cizik A U Washington SOM,
Seattle, WA.
Purpose of Study: Attributes of femoral condyle and tibial plateau geometry may play a role in the stability of the knee. We studied differences in
lateral knee geometry in ACL-injured and uninjured cohorts.
Methods Used: 185 age and activity-matched uninjured and non-contact
unilateral (uACL)- and bilateral (bACL)-injured groups were included. MRI
measurements of the articular cartilage surfaces in the mid-weight-bearing
sagittal plane of the lateral compartment evaluated tibial plateau radius of
curvature (TPr) and distal femoral radius of curvature (Fr). Femoral condyle
(FAP) and tibial plateau (TPAP) lengths were also measured. Measurements
were made by 3 observers blinded to gender and injury group.
Summary of Results: The mean lateral plateau radius of curvature, femoral
radius of curvature, and tibial plateau lengths were all significantly smaller in
ACL-injured versus non-injured groups (33.6 vs 37.4mm, p=0.003; 25.1 vs
24.2mm, p=0.01; 31.4 vs 33.2mm, p=0.004; respectively). Femoral length
was not different between ACL-injured and uninjured groups (p=0.19).
Gender Groups: Compared to uninjured males, ACL-injured males
demonstrated significantly smaller tibial plateau radius (41.1 vs. 35.3mm,
p=G0.001), femoral radius (26.7 vs. 25.5mm, p=0.002), and tibial plateau
length (35.5 vs. 33.1mm, p=G0.001). No differences in geometry were noted
between female groups.
Conclusions: Our data suggest that all females (injured or uninjured) and
ACL-injured males share a common lateral knee geometry characterized by a
shorter tibial plateau length relative to the femur and smaller (more convex)
articulating surfaces of both the proximal tibia and distal femur. It is possible
that the opposition of two highly-convex articulating surfaces may be in-
206
&
Volume 59, Number 1, January 2011
herently less stable to anterior tibial translation and rotation. These findings
also may help to explain the apparent female predilection for ACL-injury
compared to males, but suggest that this may be as much driven by geometry
as it is by gender.
421
PREOPERATIVE CT IMAGING IN REOPERATIVE
CARDIAC SURGERY
Chen WS, Wu J, Johnstone S Cardiothoracic Surgery, Kaiser Permanente,
Honolulu, HI, USA, Honolulu, HI.
Purpose of Study: Preoperative imaging is being utilized with increasing
frequency in the planning of reoperative cardiac surgery cases. This study
sought to evaluate the use of preoperative CT imaging in our reoperative
cases and its impact on operative approach and surgical outcome.
Methods Used: This is a retrospective review of all patients who underwent
reoperative cardiac surgery from 2000Y2009 at a single health maintenance
organization hospital. We evaluated operative reports of all reoperative cases
to determine if there was significant modification of surgical approach.
Modification of surgical approach was defined as incision other than median
sternotomy, femoral or axillary exposure prior to sternotomy, initiation of
cardiopulmonary bypass before sternotomy, or use of circulatory arrest. The
rate of modification of surgical approach was compared between the groups
of patients with preoperative CT imaging and without imaging. A comparison
of the observed-to-expected mortality rates was made using the Euroscore
prediction model.
Summary of Results: A total of 128 reoperative cardiac surgical cases
were performed from 2000Y2009. Preoperative CT imaging was used in 49
patients. Surgical approach was modified in 49% (24) of patients with preoperative CT imaging and only 34% (27) of patients without preoperative CT
imaging (p = 0.17). Observed-to-expected mortality rates were 1.5 and 1.28
respectively for the group with preoperative CT imaging and the group
without.
Conclusions: Patients undergoing preoperative CT imaging had a greater
rate of modification of surgical approach than those who didn’t. However, this
did not have a significant impact on the observed-to-expected mortality ratio.
While additional studies are necessary, our results indicate that the use of
preoperative CT imaging could be useful in the planning of reoperative
cardiac surgery. Preoperative CT imaging may provide information resulting
in the modification of surgical approach, possibly leading to safer surgery. We
believe it is worthwhile to consider preoperative CT imaging in high-risk
patients undergoing reoperative cardiac surgery.
422
GHRELIN ADMINISTRATION ATTENUATES THE
MYOCARDIAL INFLAMMATORY RESPONSE TO
HYPOTHERMIC ISCHEMIA/REPERFUSION
AND AFFORDS CARDIOPROTECTION
Austin EW, Ghaly Yousif N, Ao L, Fullerton D, Meng X University of
Colorado School of Medicine, Aurora, CO.
Purpose of Study: During most cardiac surgical operations, the heart must
endure the injuries of hypothermic ischemia followed by blood reperfusion.
Mechanisms of inflammation are recognized to contribute to injuries of ischemia/reperfusion (I/R). Ghrelin is an endogenous peptide, principally
produced by the gastric mucosa, and has recently been shown to have antiinflammatory actions. We hypothesized that exogenous administration of
ghrelin would reduce the myocardial injury associated with hypothermic
ischemia and blood reperfusion. In a heterotopic heart transplant model, the
purpose of this study was to examine the anti-inflammatory actions of ghrelin
in the myocardium.
Methods Used: Syngeneic mice (C3H/HeN strain) underwent heterotopic
heart transplantation. Donor hearts were stored in cold, cardioplegic solution
for 4 hours. Recipient mice received synthetic ghrelin (100 Kg/kg, iv) prior to
cervical implantation of the heart. Donor hearts were reperfused by the
recipient_s blood for 4 hours, and then harvested for analysis. Myocardial
injury was evaluated by serum cardiac troponin-I (cTn-I) levels. Levels of
monocyte chemotactic protein-1 (MCP-1), intercellular adhesion molecule-1
(ICAM-1) and monocyte infiltration were assessed by ELISA, immunoblotting and immunofluorescence, respectively. Controls underwent the same
procedure without ghrelin. Statistical analysis was performed using Student’s
t-test (pG0.05 significant).
* 2011 The American Federation for Medical Research
Journal of Investigative Medicine
&
Volume 59, Number 1, January 2011
Summary of Results: Ghrelin was cardioprotective, and this protection
was associated with significant anti-inflammatory actions. Ghrelin treated
animals exhibited less myocardial MCP-1 and ICAM-1 by 68.3% and
61.3% (both pG0.05), respectively. These findings correlated with a 58.3%
reduction of monocytic infiltrates in the myocardium (pG0.05). These antiinflammatory actions were associated with significantly reduced circulating
levels of cTn-I (pG0.05).
Conclusions: Exogenous administration of the anti-inflammatory protein,
ghrelin, was cardioprotective. It suppressed myocardial expression of MCP-1
and ICAM-1, and reduced mononuclear cell infiltration and myocardial
damage. Based on these novel findings, we conclude that ghrelin is a potent
suppressor of the myocardial inflammatory response and protects the myocardium against I/R injury.
423
AUGMENTED TENDON ACHILLES REPAIR USING
A TISSUE REINFORCEMENT SCAFFOLD:
A BIOMECHANICAL STUDY
Frizzell L, Giza E, Farac R, Williams J UC Davis, Sacramento, CA.
Purpose of Study: Missed or chronic Achilles tendon ruptures may have
muscle atrophy, tendon retraction, and a defect that must be augmented with
endogenous or exogenous materials. The Artelon\ Tissue Reinforcement
(ATR) scaffold is a readily available synthetic degradable poly(urethane urea)
material used to augment tendon repair. The objective of this study was to
compare load to failure of human cadaveric Achilles tendon repairs with and
without ATR.
Methods Used: Eighteen fresh frozen human cadaver limbs were dissected
and a rupture was simulated two cm proximal to the calcaneal insertion. The
control group of nine specimens was repaired with sutures, while the experimental group was repaired with sutures and reinforced with a tubularized
patch of ATR. Specimens were tested for ultimate load to failure in an Instron
machine after preloading to 10 N followed by cyclic loading for 20 cycles
from two to 30 N.
Summary of Results: The ultimate load to failure in the control group was
a mean of 248.1 N T 19.6 (202 to 293 at 95% CI) versus 370.4 N T 25.2 (312
to 428 at 95% CI) in the ATR group. The ultimate load to failure was 370.4 T
25.2 N (312 to 428 at 95% CI) and 248.1 T 19.6 N (202 to 293 at 95% CI) in
the experimental and control groups, respectively (p=0.0015). Creep of the
ATR augment group was 2.051 T 0.517 mm, compared to 3.126 T 1.141 mm
for the control group (p=0.026).
Conclusions: ATR is a readily available material that can be used to augment chronic or acute Achilles tendon ruptures. It is less bulky than currently
available allograft or xenograft alternatives and provides a statistically considerable improvement in load to failure when compared to control specimens
in a cadaver model. This finding may allow for development of more aggressive rehabilitation techniques following chronic Achilles tendon repairs.
Western Regional Meeting Abstracts
MATLAB Accuracy: Calculation of relative marker motion yielded
nonsense data with very large noise signal at small ranges of motion
(È6300% at 0.005[), but became much more accurate at large displacements
(1.3% at 1.500[). Accuracy remained poor even at large displacements in the
Y-axis (A/P), with È13% error, but was comparable between X- and Z-axes.
Signal to noise exceeded 4:1 in the axis of interest at 0.020[, but noise in
other axes was still significant.
Calibration Error Tolerance: Recalculation of displacement data using
calibration footage from alternate camera position and angle showed negligible differences in accuracy.
Conclusions: Existing automatic tracking algorithms match or exceed
human tracking of markers, where the proposed system performs with a fixed
absolute error of under È0.005[ at a focal distance of È36[. At displacements less than 0.020[, point-to-point position error is poorly tolerated by the
MATLAB algorithm. However, the results do show the high tolerance of the
system to environmental perturbations via replication of output data amidst
minor changes in camera position and angle.
425
SEVERE PRE-LIVER TRANSPLANT CORONARY ARTERY
DISEASE ASSOCIATED WITH SIGNIFICANT WORSENING
OF LATE POST-TRANSPLANT SURVIVAL
Simon N1, Mangus S2, Kinsella S1, Fridell J2, Vianna R2, Quirk C1, Wilkes
K1, Tector J2 1Indiana University School of Medicine, Indianapolis, IN and
2
Indiana University School of Medicine, Indianapolis, IN.
Purpose of Study: Coronary artery disease (CAD) is an important risk
factor for perioperative complications and worse clinical outcomes in patients
undergoing liver transplantation (LT). This study reviews the pre transplant
cardiac history of a large number of LT recipients and evaluates post transplant outcomes for those with a history of severe CAD.
Methods Used: A complete cardiac history was available for 1122 of 1170
LT recipients (96%) from 2001 to 2010. Patients were categorized into 3
study groups, based upon a history of (1) no previous cardiac intervention, (2)
a Bremote[ (912 months pre transplant) intervention, or (3) Brecent[ (within
12 months of transplant) cardiac intervention. Intervention included either
surgical revascularization, angioplasty or placement of a coronary artery
stent. A Cox proportional hazards model was constructed using direct variable entry to evaluate long-term post-transplant patient survival.
Summary of Results: There were 59 patients with a history of pre transplant cardiac intervention (5.2%), 3.7% recent and 1.5% remote. Median
follow up for this cohort was 59 months. Rates of 90-day post LT MI, stroke
and DVT did not differ among the 3 study groups. The Cox patient survival
curve is shown in Figure-1.
Conclusions: Patients with previous cardiac intervention, both recent and
remote, had significantly worse long term post LT survival. However, these
groups did not have significantly worse perioperative morbidity.
424
VALIDATION OF REAL-TIME, LOW-COST STEREO
VIDEOGRAPHY IN MEASURING 3D SPINE MOTION
Namperumal S1, Leung E2, Curtiss S1, Roberto R1 1UC Davis School of
Medicine, Sacramento, CA and 2UC Davis, Davis, CA.
Purpose of Study: A low-cost spine motion measurement system is validated for accuracy and resolution to aid in the development of therapeutic,
motion-sparing fixation hardware.
Methods Used: Stereo high-resolution digital video was taken of precise
linear motions of LED triad markers in a 3D measurement space. Calculation
of marker positions was performed using SIMI motion analysis software
(SIMI Reality Motion Systems, Germany) using both automatic and manual
tracking of marker motions. A previously validated MATLAB algorithm was
then used to calculate the relative motion between markers. Error calculations
were made for both SIMI and MATLAB displacement data to validate the
accuracy of 3D motion measurement in the proposed system.
Summary of Results: Tracking: Noise generated at rest in automatic
tracking (29%Y81%, mean position of 0.0019[) was comparable to that of
manual tracking (38%Y87% of 0.0026[), and accuracy of automatic tracking
was comparable to or exceeded that of manual tracking (mean error of
0.0003[ to 0.0032[) over absolute displacements from 0.005[ to 1.000[.
Accuracy was comparable in all axes of 3D motion.
* 2011 The American Federation for Medical Research
Figure 1. Cox proportional hazaeds model for post LT survival.
207
Western Regional Meeting Abstracts
Journal of Investigative Medicine
426
SONOGRAPHIC EVALUATION OF THE PERONEUS
LONGUS AND BREVIS TENDONS: ANATOMICAL
RELATIONSHIPS AT REST AND UNDER STRESS
Chen LE1, Hager NA2,3 1University of Washington School of Medicine,
Seattle, WA; 2University of Washington Medical Center, Seattle, WA and
3
University of Washington Medical Center, Seattle, WA.
Purpose of Study: Ultrasound has been considered a valuable imaging
modality to assess superficial tendons due to its high resolution, noninvasiveness, and cost effectiveness. The normal locations of the peroneus longus
and brevis tendons at rest and under stress have not been well characterized in
the literature. It is hypothesized that the tendon position changes are correlated with the various provocative maneuvers and the stress applied to the
tendons. The goal of the study is to identify the particular provocative maneuver of an ankle that demonstrates the greatest change in tendon positions.
Practitioners could then rely on this maneuver as their primary assessment
method.
Methods Used: The sample population consisted of 17 asymptomatic
subjects (34 ankles) with no history of ankle pathology. Inversion, eversion,
plantarflexion, and dorsiflexion were performed, first passively without resistance then actively against resistance. Transverse dimensions were imaged.
The changes in the positions of the peroneal tendons at rest and under stress
were determined by the displacement angle of the peroneus brevis tendon
with respect to the peroneus longus tendon.
Summary of Results: At the resting position, 65% of the samples showed
the peroneus brevis tendon off-center lateral to the peroneus longus tendon.
No statistically significant difference was seen in the tendon displacements
among four passive motions. Under stress, eversion combined with dorsiflexion demonstrated an averaged displacement of 66- T 12- compared with
the resting position, and this result does not show statistically significant
difference from pure eversion (56- T 6.9-). Inversion against resistance
showed the least position change (25- T 3.5-) from the tendons at rest.
Conclusions: Without resistance, the tendons do not show significant differences in positions among various passive maneuvers. The greatest displacements of the relative tendon positions were noted during eversion
combined with dorsiflexion against resistance. These findings suggest that
the optimal provocative test maneuver for assessing the status of the peroneal
tendons is eversion and dorsiflexion of the foot against resistance.
427
PREDICTORS OF COMPLICATIONS IN CHILDREN
WITH DUPLICATED RENAL COLLECTING SYSTEMS
Spelliscy C, Kiddoo D University of Alberta, Edmonton, AB, Canada.
Purpose of Study: 1. To identify which factors are correlated with urinary
tract infections (UTIs) and decreased renal function in children with antenatally
diagnosed duplicated renal collecting systems (DRCS). 2. To provide suggestions as to early management i