2011 ADA Posters 1261-2041.indd - Diabetes

Integrated Physiology/ 

Obesity 

POSTERS 

1622-P 

Hyperglycemia Mediated Accelerates Aging in Vascular Endothelial 

Cells 

ROKHSANA MORTUZA, SHALI CHEN, BIAO FENG, SUBRATA CHAKRABARTI, 

London, ON, Canada, Lindon, ON, Canada 

Glucose-induced endothelial cell dysfunction is a main factor causing 

tissue damage in the eye, kidney, heart and nerve in Diabetes. We 

hypothesized that hyperglycemia induced oxidative stress causes an 

accelerated aging process and that such process is mediated through 

alterations of silent information regulator proteins or sirtuins(SIRTs) through 

epigenetic mechanisms. 

We investigated glucose induced aging and alterations of SIRTs in two 

types of endothelial cells, human umbilical vein endothelial cells(HUVECs) 

and human microvascular endothelial cells(HMECs). The cells were grown 

and propagated in high glucose (25 mmol, HG) and low glucose(5 mmol, NG). 

Cells were observed daily and images taken to analyze their rate of growth 

and confl uency. At each passage subculture, some cells were collected 

for analysis of SIRTs, excision repair cross complimenting 1 and 4(ERCC1, 

ERCC4) by RT-PCR. Cells from each passage were stained for senescence 

activated beta galactosidase(SA-βgal), a known biomarker of cellular aging. 

Morphometric analyses were performed. 

Both types of cells grew faster in NG compared to HG. When exposed to 

HG, both cell types showed evidence of early senescence as evidenced by 

increased SA-βgal expression. The replicative capacities of these cells were 

diminished and they developed an irregular and hypertrophic phenotype. 

Interestingly, we found that HMEC cells started showing such changes after 

passage 1 in HG(compared to passage 4 in NG), whereas such changes were 

seen in HUVECs at passage 4 in HG(compared to passage 7 in NG). Almost 

all HMECs in HG at passage 5(compared to only 1.5% in NG) and HUVECs 

in HG at passage 11(compared to 61.4% in NG) showed β-gal positivity. 

Quantitative RT-PCR analyses showed this aging process was associated 

with decreased SIRT 1, 2, 5 expressions in both cell lines. ERCC1, ERCC4 

mRNA expression was found to be decreased in HMEC but increased in 

HUVEC in earlier passages. Such changes were prevented by treatment with 

resveratrol. 

Data from this study suggests that hyperglycemia leads to accelerated 

aging in the endothelial cells. Furthermore, such aging process is mediated 

through SIRTs, ERCC1 and 4 and varies depending on the cell type. 

1623-P 

Transcription Factor mafB and a Functional Link to E-cadherin 

Expression in β Cell Differentiation 

MARIKO TSUCHIYA, KEN TSUCHIYA, KOSAKU NITTA, ATSUSHI MAEDA, Tokyo, 

Japan 

It is essential to clarify the mechanism by which endocrine cells including 

β-cells differentiate or transform during development or regeneration in 

the pancreas. First, we identifi ed a few cells in the ductal epithelium of 

the pancreas of newborn that stained positive for insulin and mafB and 

negative for E-cadherin, whereas almost all the surrounding duct cells 

stained positive for E-cadherin, suggesting that there may be cross-talk 

between mafB and E-cadherin. The purpose of the present study was to 

elucidate the consequences of E-cadherin and mafB expression in relation 

to induction of ductal epithelial cell (β-cell progenitor) differentiation into 

endocrine cells by mafB. We used the technique to intravenously inject 

mice and transfect cultured cells with synthetic mafB-siRNA and analyzed 

the resulting alterations in gene and protein expression level by real-time 

PCR or immunohistochemistry. To observe the precise mechanism of this 

process, we suppressed mafB gene expression to 40% of the control level 

in AsPc1 cells, a cell line derived from a pancreatic carcinoma in which 

maf expression has been confi rmed by real-time PCR. Suppression of mafB 

expression by siRNA in AsPC1 cells resulted in upregulation of mRNA (1.4 

fold) and immunostaining intensity (1.4 fold) of E-cadherin. Expression of 

α-catenin (1.2 fold), which is indispensable for cadherin-catenin-complexmediated 

cell adhesion, was also increased, and β-catenin was increased 

by 3.5 fold and GSK3 was slightly suppressed (0.8 fold), suggesting that 

mafB may regulate ductal epithelial progenitor differentiation by expression 

of the cadherin-catenin complex in the WNT signaling pathway. In the in 

vivo study mafB-siRNA treatment in the mouse pancreas resulted in similar 

changes. Moreover, mafB suppression downregulated expression of γ-actin 

by the results of microarray profi ling. Taken together, the fi ndings in this 

study suggested that mafB, whose expression is modifi ed by other mafs or 

other genes, even in stress or metabolic conditions in the mature pancreas 

may regulate the E-cadherin-catenin complex in the WNT signaling pathway, 

resulting in cell adherence and endocrine cell differentiation. 

For author disclosure information, see page 785. 

INTEGRATED PHYSIOLOGY—INSULIN CATEGORY SECRETION IN VIVO 

A442 

1624-P 

Unique Cellular Responses of Adult Blood-Derived Endothelial Progenitor 

Cells and Mature Endothelial Cells to High Glucose 

EMILY KEATS, ZIA A. KHAN, London, ON, Canada 

Diabetes leads to dysfunction of selected organ systems, and vascular 

endothelial cell (EC) dysfunction and loss is the key initiating and perpetuating 

step in the development of secondary diabetic complications. A number 

of studies have investigated the effect of diabetes on non-vasculogenic 

precursor cells (also known as early endothelial progenitor cells). However, a 

comparative study investigating the effects of high levels of glucose on the 

proliferative/vasculogenic endothelial progenitor cells (EPCs; late EPCs) and 

mature ECs is lacking. We herein determined the response of adult bloodderived 

EPCs and mature ECs to high levels glucose. 

Mononuclear cells (MNCs) from normal adult blood were successfully 

differentiated into ECs and characterized through our established protocol. 

Mature ECs were obtained from neonatal foreskin. Cells were then subject to 

5 mmol/L or 25 mmol/L glucose and cellular activity assays were performed. 

We further investigated the mechanisms of increased oxidative stress in 

these EC types. 

Blood-derived EPCs were characterized by fl ow cytometry, cell staining, 

and RT-PCR. The progenitor phenotype was confi rmed by our recently 

discovered endostatin response assay. Following characterization, the 

cells were exposed to high levels of glucose for 6 and 12 days. Our results 

show that short term exposure (6-day) to glucose increased endothelin-1 

while decreasing the endothelin receptors in both EPCs and mature ECs. 

Interestingly, long-term exposure of cells to glucose (12 days) increased 

endothelin receptors in mature ECs but not EPCs. These results suggest 

that mature ECs are more susceptible to the adverse effect of high glucose. 

Our results also show high glucose-induced E-selectin expression in EPCs 

suggesting an angiogenic phenotype which was not evident in mature ECs. 

Lastly, our data illustrate NADPH oxidase was increased in mature ECs but 

remained unaltered in the EPCs. 

In conclusion, our results show differential response of adult blood EPCs 

and mature ECs to high levels of glucose and evidence for the therapeutic 

potential of vasculogenic EPCs. 

Supported by: Canadian Diabetes Association and the Lawson Health Research 

Institute 

INTEGRATED PHYSIOLOGY—INSULIN SECRETION 

IN VIVO 

[See also: Presidents Posters 451-PP to 452-PP, page A125.] 

Guided Audio Tour: Insulin Secretion—In Vivo Physiology (Posters 1625-P 

to 1634-P), see page 13. 

& 1625-P 

Short-Term Administration of the Atypical Antipsychotic Olanzapine 

Induces Meal-Specifi c Impairments in Glucose and Lipid Metabolism, 

Independent of Weight Gain or Psychiatric Disease 

KAREN L. TEFF, MICHAEL R. RICKELS, JOANNE GRUDZIAK, KARL RICKELS, Philadelphia, 

PA 

The atypical antipsychotics are associated with an increased incidence 

of diabetes. The mechanisms underlying these metabolic defects are not 

understood, although it has been speculated that the initiating pathophysiology 

is weight gain, secondary to centrally-mediated increases in appetite. Our 

objectives were to determine if olanzapine administration impairs insulin 

sensitivity and post-prandial responses to the physiologically-relevant stimulus 

of a meal, independent of weight gain or psychiatric disease. Healthy male 

subjects (BMI=69.8+2.6 Kg/m 2 ) were admitted into the CTRC for 12-days. 

Activity was maintained for each subject at a level matched to their free 

living levels. On Days 1 and 3 in a randomized order, subjects underwent a 600 

kcal mixed nutrient meal challenge (n=10) and a euglycemic, hyperinsulinemic 

clamp (7/10 subjects). The meal was labeled with [1- 13 C] glucose to monitor 

glucose appearance and 6,6-[ 2 H 2 ]-glucose was infused to measure glucose 

disposal and endogenous glucose production during both the meal challenge 

and the clamp. Subjects were then administered olanzapine (5 mg/d, Days 4-5; 

10 mg/d, Days 6-12). On Days 10 and 12, the metabolic tests were repeated. 

Weight remained stable throughout the study (69.8+2.6,pre:70.1+2.9 Kg/ 

m 2 , post-olanzapine) and no increase in food intake was found. Olanzapine 

increased post-prandial glucose (area under the curve: pre, 4363+1981 vs 

post, 6579+1838, P

signifi cant decreases in glucose disposal were found during the meal and the 

clamp (P


Obesity 

POSTERS 

Despite of a markedly enhanced GLP-1 response there was no improvement 

in insulin secretion but an unexpected increase in glucagon levels during the 

OGTT shortly after gastric bypass surgery. While gastric bypass surgery in 

the short-term appears to leave beta cell function unaffected it obviously 

exerts a strong stimulating effect on alpha cell function. Collectively, our 

data speak against a major contribution of incretin effects to the early 

improvement in glucose metabolism after gastric bypass surgery which 

appears to rely primarily on enhanced insulin sensitivity. 

DM C 

pre-operative post-operative pre-operative post-operative 

GLP-1 AUC (pg/ml/min) 0.56±0.34 2.61±0.96 0.46±0.35 3.58±1.72 

Glucagon AUC (pg/ml/min) 11.27±3.10 52.20±16.19 10.11±4.22 53.36±12.14 

Supported by: EFSD/MSD Clinical Research Grant 2009 

& 1629-P 

Impaired Insulinotropic Effect of Both GLP-1 and GIP Can Be Induced 

in Healthy Normal Glucose Tolerant Subjects 

KATRINE B. HANSEN, TINA VILSBØLL, JONATAN I. BAGGER, JENS J. HOLST, 

FILIP K. KNOP, Glostrup, Denmark, Gentofte, Denmark, Copenhagen, Denmark 

The insulinotropic effect of the incretin hormones glucose-dependent 

insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) is 

impaired in patients with type 2 diabetes. It remains unclear whether this 

impairment can be induced temporarily in healthy subjects by a short period 

of glucose homeostatic dysregulation. 

We aimed to investigate the insulinotropic effect of GIP and GLP-1 

compared to placebo (saline) before and after 12 days of glucose homeostatic 

dysregulation in healthy subjects. 

The insulinotropic effect was measured using hyperglycemic clamps and 

infusion of physiological doses of GIP, GLP-1 or saline in 10 healthy Caucasian 

males (age: 25±1 years (mean±SEM); BMI: 23±1kg/m 2 ; HbA 1 c: 5.4±0.1%; no 

family history of diabetes) before and after intervention using high calorie 

diet, sedentary lifestyle and administration of prednisolone (37.5 mg/day) 

for 12 days. 

The intervention resulted in insulin resistance (IR) according to the 

homeostatic model assessment (1.2±0.2 vs 2.6±0.5, P=0.01) and glucose 

tolerance deteriorated as assessed by the area under curve (AUC) for 

plasma glucose during 75 g-OGTT (730±30 vs. 846±57mM×2h, P=0.021). The 

subjects compensated for the induced IR by signifi cantly increasing their 

post intervention insulin responses during saline infusion by 2.9±0.5 fold; 

signifi cantly (P=0.001) more than during GIP or GLP-1 infusions (1.78±0.3 and 

1.38±0.3, P=NS). 

These data show that impairment of the insulinotropic effect of both GIP 

and GLP-1 can be induced in healthy male subjects without risk factors for 

type 2 diabetes. This fi nding indicates that the impaired insulinotropic effect 

of the incretin hormones observed in type 2 diabetes is a consequence 

of insulin resistance and glucose intolerance rather than a primary event 

causing type 2 diabetes. 

Supported by: EFSD/Novartis Grant 

& 1630-P 

Pioglitazone (PIO) Does Not Enhance Glucose-Dependent Insulinotropic 

Peptide (GIP) Stimulated Insulin Secretion in Subjects with 

Well-Controlled T2DM 

WILLIAM G. THARP, OLGA SIDELEVA, DARIUSH ELAHI, COURTNEY LEDGER, 

RHONDA MAPLE, RICHARD PRATLEY, Burlington, VT, Baltimore, MD 

Dysregulation of incretin action contributes to hyperglycemia in Type 

2 diabetes(T2DM). GIP stimulated insulin secretion(GIP-SIS) is markedly 

blunted, possibly due to down-regulation of GIP receptor in β-cells by chronic 

hyperglycemia. The promoter of GIP receptor contains a PPAR response 

element. We hypothesized that treatment with PIO, a potent PPARγ agonist, 

would improve GIP-SIS in subjects with T2DM. Twenty four subjects (12M/12F, 

age 58.7+10.6, BMI 33.4+8.1) with well controlled T2DM (A1c

(t=5, t=10, t=15, t=30, t=60) and following cessation of stimulation (t=90 

and t=120 minutes) to determine blood glucose and plasma insulin levels. To 

exclude that the effects on glucose metabolism were stress-induced we also 

measured plasma corticosterone levels. 

In the 200 μA stimulated animals, plasma insulin levels signifi cantly 

increased to 122% of base-line at onset of stimulation (t=5) compared to 

sham stimulated animals whereas blood glucose values showed a slight (4%) 

increase during the fi rst 15 min of stimulation. In contrast, stimulation at 

lower intensity (100 μA) did not affect either measure. Plasma corticosterone 

levels were not signifi cantly changed as compared to the sham and 100 μA 

conditions and thus can not explain changes in insulin secretion or the slight 

glucose rise. 

These data suggest a role for the NAc in the regulation of glucose 

metabolism and we hypothesize that the NAc controls the pancreas and 

thereby insulin secretion via the central nervous system. 

Supported by: ZonMW 

& 1633-P 

Synergism of Dopamine Agonist Plus GLP-1 Analog Therapy on 

Improvement of Glucose Intolerance in Syrian Hamsters 

MICHAEL EZROKHI, SHUQIN LUO, YELENA TRUBITSYNA, ANTHONY H. CIN- 

COTTA, Tiverton, RI 

Previous studies have demonstrated that timed daily treatment of 

bromocriptine, a dopamine D2 receptor agonist, to insulin resistant animals 

and humans improves postprandial insulin sensitivity. Such treatment 

reduces post-meal or post-glucose challenge levels of glucose while 

simultaneously markedly reducing hyperinsulinemia. GLP-1 analogs such 

as exendin-4, are known to enhance glucose – mediated insulin secretion 

and reduce glucagon secretion. Therefore, the possibility exists of a positive 

interaction between such a postprandial insulin sensitizing mechanism 

with such an enhanced postprandial insulin secretory response to improve 

glucose intolerance. 

To test such a postulate, glucose intolerant insulin resistant Syrian 

hamsters weighing 200 grams were treated with either vehicle (N=20) or 

bromocriptine (4 mg/kg; N=20) for 2 weeks and then each group was further 

subdivided into 2 groups and treated with either vehicle or exendin-4 (4 µg/ 

kg) just prior to being subjected to a glucose tolerance test (GTT) (3g/kg). 

Relative to the vehicle/vehicle (control), bromocriptine markedly reduced the 

insulin area under the curve (AUC) by 39% (from 1311 ± 177 to 796 ± 119 

ng*min/ml) (P=0.03) while reducing the glucose AUC by 21% (from 42576 ± 3889 

to 33752 ± 2763 mg*min/dl) (P


Obesity 

POSTERS 

1637-P 

Delayed Insulin Secretory Responses Are Seen in Patients with 

Long Duration of Type 1 Diabetes and Residual Insulin* 

JENNIFER SHERR, LINDA RINK, KEVAN HEROLD, New Haven, CT 

Calculation of insulin secretory rates (ISR) can provide quantitative and 

qualitative information of insulin secretion during a mixed meal test (MMT). 

We previously reported that some patients with type 1 diabetes (T1D) have 

a delayed secretory response during a MMT compared to non-diabetic 

controls and that their decline in insulin secretion was less than patients 

with T1D who showed an early secretion profi le (i.e. @< 45min). To determine 

if there is a relationship between disease duration and ISRs, data derived 

from MMT were analyzed in 1) 20 subjects with T1D diagnosed < 1 year 2) 

12 subjects with T1D 2-5 years duration, 3) 21 subjects with T1D >5 years, 

4) 38 non-diabetic controls. All subjects with T1D 5 

years subjects had undetectable c-peptide levels and were excluded from 

further analysis. Chronobiological Series Analyzer (CSA) software was used 

to calculate ISR by deconvolution of C-peptide using a two-compartment 

model for hormone clearance. Parameters used accounted for the subject’s 

age, sex, height, and diabetes status. Total ISR area under the curve (AUC) 

and time of peak insulin secretion was calculated for each subject. There 

was a decline in total insulin secretion with duration of T1D (p

As part of a randomized, placebo-controlled study of salsalate (SAL) 

and glucose metabolism in patients with IFG and IGT, we investigated 

whether treatment with a high dose of SAL for 3 months would modulate 

I and C-peptide (C) responses to a 75-g oral glucose tolerance test (OGTT), 

and metabolic clearance of insulin (MCI) during both OGTT and euglycemichyperinsulinemic 

clamp (EHC). Present analyses include data from 49 subjects 

(24 SAL/25 placebo) with follow-up OGTTs performed 2 and 3 months (Mo 

2 and Mo 3) after randomization and from 57 subjects (28 SAL/29 placebo) 

with follow-up EHC performed at study-end. 

SAL reduced fasting glucose (Gluc) (11%, p=0.0001), did not change fasting I, 

but reduced fasting C (31% Mo 2, p


Obesity 

POSTERS 

infusion and 30 min GIP infusion (2 pmol/kg/min). The acute insulin response 

to glucose (AIRg) and insulin sensitivity (Si) were derived from the Minimal 

Model, β-cell sensitivity to glucose (GSIS) was calculated as the slope of 

insulin to glucose during the glucose infusion, and GIP-SIS as the increment 

in insulin secretion above GSIS during the GIP infusion. Twenty-four subjects 

were studied (Table). There were no differences in A1C between treatments, 

but BMI was higher in the MET group than D+E (p=0.033) and higher in 

women than men (38.4±8.6 vs 28.4±3.4, p=0.001). AIRg, Si, and GIP-SIS did 

not differ by treatment, but GSIS was ∼3-fold higher in subjects on MET after 

controlling for AIRg and BMI (p=0.044, r2 =0.70). GIP-SIS correlated with AIRg 

(p

WT mice compared to islets of saline infused WT mice. In contrast, islets of 

TG mice subjected to 48h elevation of FFA had no signifi cant impairment in 

β-cell function compared to islets of saline infused mice (Insulin Release in 

pmol/islet/h WT Saline: 0.27±0.03; WT Oleate: 0.14±0.03, p


Obesity 

POSTERS 

Conscious hyperinsulinemic euglycemic clamp was performed in 

a separate set of Ob/Ob mice or lean controls. Si, AIRg, Sg and DI were 

calculated by Minimal Model analysis of FSIVGTT data. Validating FSIVGTT 

against the gold standard euglycemic clamp, the FSIVGTT-derived Si of lean 

vs. Ob/Ob mice (10.6±.19 vs. 1.3±0.5 mU/ml/min, p=0.001) was equivalent to 

clamp glucose infusion rates (38.7±0.8 vs. 6.0±0.2 mg/kg/min, p

expression. These fi ndings suggest that TLR4 in Kupffer cells mediates the 

progression from simple steatosis to infl ammation, partly by inducing the 

production of ROS, thereby leading to the activation of XBP-1. 

Supported by: Hong Kong Research Grant Council (HKU 5/CRF/08 and HKU 

2/07C) 

& 1656-P 

Liver-Specifi c Deletion of JAK2 Leads to Profound Fatty Liver but 

Suppression of Infl ammation and Protection Against Systemic Insulin 

Resistance 

RUBÉN GARCÍA, SALLY YU SHI, DIANA CHOI, ROBIN E. DUNCAN, SHUN YAN LU, 

STEPHANIE A. SCHROER, ERICA P. CAI, CHRISTINE TANG, ADRIA GIACCA, MARK 

NAPLES, MARK J. DEKKER, KAY-UWE WAGNER, KHOSROW ADELI, RICHARD P. 

BAZINET, MINNA WOO, Toronto, ON, Canada, Omaha, NE 

Non-alcoholic fatty liver disease (NAFLD) is considered to be the hepatic 

manifestation of the metabolic syndrome, a well-recognized precursor for 

many chronic diseases including type 2 diabetes, cardiovascular diseases 

and some cancers. NAFLD is a disease spectrum hypothesized to be 

induced by a two-step process: the fi rst hit, steatosis, sensitizes the liver 

to the infl ammatory damage induced by a second pathogenic insult, which 

promotes steatohepatitis. The exact pathogenesis of NAFLD, however, is 

not well understood. The Janus kinase-signal transducers and activators of 

transcription (JAK-STAT) pathway is involved in both infl ammatory signalling 

and in lipid homeostasis in the liver and may contribute to the pathogenesis 

of NAFLD. To investigate the role of hepatic JAK2, an essential player in 

the JAK-STAT pathway, we generated mice with deletion of jak2 specifi cally 

in hepatocytes (L-JAK2 KO) using the cre-loxP system. L-JAK2 KO mice 

developed spontaneous hepatosteatosis early in life, similar to liverspecifi 

c growth hormone receptor- and STAT5-defi cient mice. However, in 

contrast to these mice, L-JAK2 KO mice did not develop systemic insulin 

resistance or glucose intolerance. Interestingly, the fatty acid composition 

of liver triglycerides showed, in addition to increased palmitate, a persistent 

predominance of the less toxic oleate, which was accompanied by reduced 

hepatic infl ammation in L-JAK2 KO mice. Even when challenged with a 

prolonged high fat diet, these mice were completely protected against 

systemic infl ammation and insulin resistance, resulting in improved glucose 

tolerance. Together, our fi ndings indicate a critical and unique role of hepatic 

jak2 in the regulation of fatty acid metabolism and infl ammation, leading 

to a complete halt in NAFLD progression and development of systemic 

insulin resistance in mice lacking JAK2 in hepatocytes. Targeting the JAK- 

STAT pathway may therefore provide a novel therapeutic strategy for the 

treatment of type 2 diabetes. 

Supported by: CIHR; Ibercaja Foundation (Spain); BBDC-Novo Nordisk Studentship 

& 1657-P 

Liver-Specifi c Ablation of PPARγ Reduces Hepatic Mitochondrial 

β-Oxidation and Lipid Accumulation, but Aggravates High Fat Diet- 

Induced Insulin Resistance 

ANISHA A. GUPTE, LAURIE J. MINZE, JESSICA R. WILES, MARTINEZ JESSICA, 

ALAN R. COLLINS, CHRISTOPHER J. LYON, WILLA A. HSUEH, Houston, TX 

Fatty liver is thought to contribute to insulin resistance and diabetes. 

Although peroxisome proliferator-activated receptor-γ (PPARγ) activation 

can increase de novo lipogenesis in the liver during high fat diet (HFD), we 

found that activation increases β-oxidation, improves liver mitochondrial 

function and decreases liver fat accumulation (Gupte A, et al, Hepatology, 

2010). However, very little is known about the role of liver-PPARγ and its 

effects, if any, upon liver metabolism. We thus generated mice with liverspecifi 

c PPARγ knockout (L-PPARγKO), and found that L-PPARγKO mice 

fed obesogenic HFD for 3 months had 50% the liver fat of wild type (WT) 

littermates. However, L-PPARγKO had elevated fasting insulin vs. WT mice 

and impaired insulin sensitivity when assayed by insulin tolerance test, 

but no differences in plasma triglycerides, phospholipids or free fatty acid 

levels. Insulin-stimulated Akt phosphorylation was suppressed in liver, fat 

and skeletal muscle of WT mice on HFD vs. chow, but was increased in liver, 

unchanged in fat, and reduced in skeletal muscle of HFD-fed L-PPARγKO mice 

vs. WT mice. Thus, L-PPARγKO mice have skeletal muscle insulin resistance, 

but decreased liver fat and improved liver insulin sensitivity. Mitochondrial 

function analyses revealed signifi cant suppression of β-oxidation in the 

livers of L-PPARγKO mice, but there was reduced expression of lipogenic 

genes (e.g. fatty acid synthase) corresponding with reduced hepatic fat 

accumulation, suggesting that liver-PPARγ directly regulates hepatic lipid 

synthesis and metabolism. Surprisingly, the PPARγ agonist rosiglitazone 

restored β-oxidation in L-PPARγKO mice on HFD to that seen in WT mice. 

These data suggest: 1) liver PPARγ directly regulates hepatic lipid synthesis 

ADA-Funded Research 

& Guided Audio Tour poster 

INTEGRATED CATEGORY PHYSIOLOGY—LIVER 

A451 

and mitochondrial β-oxidation of fatty acids, 2) PPARγ effects outside the 

liver also impact hepatic β-oxidation, and 3) liver PPARγ may exert signifi cant 

effects upon skeletal muscle insulin sensitivity, even in the absence of 

liver fat. We conclude that liver PPARγ may have important therapeutic 

implications for liver complications found in obesity and diabetes. 

& 1658-P 

Rat Krueppel-Like Factor 10 (Klf-10) Gene Expression Is Regulated 

by the Carbohydrate Response Element Binding Protein ChREBP in 

Primary Rat Hepatocytes 

KATSUMI IIZUKA, YUKIO HORIKAWA, REIKO TOMITA, TETSUYA SUWA, JUN 

TAKEDA, Gifu, Japan 

We previously reported that the carbohydrate response element binding 

protein ChREBP plays an important role in the regulation of glucose and lipid 

metabolism by regulating hepatic glycolytic and lipogenic gene expression. 

We then established a mouse model infected by an adenovirus expressing 

dominant active ChREBP (daChREBP). Using the DNA microarray technique, 

we identifi ed the Krueppel-like factor 10 (Klf-10) as a candidate for one of 

the ChREBP target genes. Recently, Klf-10 has been identifi ed as a circadian 

transcriptional regulator that links the molecular clock to energy metabolism 

in the liver. Here we investigate whether ChREBP directly regulates Klf-10 

gene expression in rat primary hepatocytes and the pancreatic beta cell 

line INS-1E. Klf-10 mRNA is ubiquitously expressed and highly expressed 

in muscles and the liver. The hepatic Klf-10 mRNA in ob/ob mice is about 

2-fold higher than that in C57BL/6 mice. In rat primary hepatocytes and 

INS-1E cells, glucose stimulation and overexpression of daChREBP induces 

klf-10 mRNA in a dose-dependent manner. Consistent with these fi ndings, 

overexpression of dominant negative Mlx inhibits glucose induction of liver 

type pyruvate kinase (Lpk) and fatty acid synthase (Fasn) mRNA. A deletion 

study of pGL3 vector with rat KLF-10 promoter and a CHIP assay against anti- 

ChREBP antibody demonstrated that the carbohydrate response element 

(ChoRE) is located between -200 and -100 bp in the rat Klf-10 gene promoter. 

In addition, adenoviral overexpression of Klf-10 partly inhibits glucose 

induction of ChREBP target genes (Lpk and Fasn) in primary hepatocytes. In 

conclusion, these fi ndings suggest that ChREBP directly regulates Klf-10 gene 

expression at the transcription level and that Klf-10 also weakly contributes 

to regulating ChREBP transactivities. Crosstalk between ChREBP and Klf-10 

is involved in the regulation of the lipogenic pathway. 

& 1659-P 

Role of Sdf2l1, a Novel ER Stress-Related Protein, in the Regulation 

of Hepatic Insulin Sensitivity 

TAKAYOSHI SASAKO, KOHJIRO UEKI, MITSURU OHSUGI, NAOTO KUBOTA, 

KAZUYUKI TOBE, TAKASHI KADOWAKI, Tokyo, Japan, Toyama, Japan 

Dys-regulation of dynamic metabolic changes in liver during the transition 

between fasting and feeding leads to metabolic disorders. To further 

elucidate these changes, we compared global gene expressions in livers 

of B6J mice in 24h-fasted and 6h-refed states by microarray analysis. 

Interestingly several ER stress-related genes were up-regulated by 

refeeding. Among them, we focused on Sdf2l1, a component of chaperone 

complexes, showing the second largest increase of all the genes. Indeed 

hepatic expression of ER stress markers as well as Sdf2l1 was elevated 

prominently and transiently by refeeding in B6J mice. In order to explore the 

role of Sdf2l1 in response to ER stress, we performed promoter assay and 

ChIP assay, revealing that Sdf2l1 was regulated by Xbp1 and ATF6, despite 

absence of conventional ERSE in the promoter region. Since ER stress is 

thought to be a pivotal regulator of hepatic insulin sensitivity, to assess the 

role of Sdf2l1 in this context, we knocked down hepatic Sdf2l1 expression 

by adenovirus-mediated RNA interference. Knocking down of Sdf2l1 in B6J 

mice resulted in greater elevation of ER stress markers after refeeding, while 

it impaired glucose tolerance, attenuated insulin signaling after refeeding, 

and increased triglyceride contents. These prompted us to hypothesize that 

dys-regulation of Sdf2l1 might contribute to insulin resistance in obesity. 

Indeed in livers of db/db mice, a model of obesity and diabetes, Sdf2l1 was 

down-regulated, accompanied by decreased binding of Xbp1, but not ATF6, 

with the promoter region. By contrast, adenovirus-mediated restoration of 

Sdf2l1 in livers of db/db mice improved glucose tolerance, insulin signaling 

after refeeding, and fatty liver. These data suggest that feeding induces ER 

stress in liver even under the physiological condition, while Sdf2l1 regulated 

by Xbp1 and ATF6 may switch off the ER stress response, maintaining 

hepatic insulin sensitivity after feeding. On the other hand, suppression of 

hepatic Sdf2l1 expression in obesity may contribute to the development of 

systemic insulin resistance, providing the possibility of Sdf2l1 to be a novel 

therapeutic target of obesity-induced diabetes. 



Obesity 

POSTERS


Obesity 

POSTERS 

& 1660-P 

FoxO1 Regulates Sterol Regulatory Element Binding Protein-1c Gene 

Expression 

XIONG DENG, CHANDRAHASSA YELLATURU, WENWEI ZHANG, INSUG O-SULLIVAN, 

JAMES B. WILLIAMS, MARSHALL B. ELAM, EDWARDS A. PARK, RAJENDRA RAG- 

HOW, TERRY UNTERMAN, Memphis, TN, Chicago, IL 

Induction of lipogenesis in response to insulin is critically dependent on the 

transcription factor, sterol regulatory element-binding protein-1c (SREBBP- 

1c). FoxO1, a forkhead box class-O transcription factor, is an important 

target of insulin action, but its role in lipid metabolism has not been clearly 

defi ned. We conducted studies to examine the effects of FoxO proteins on 

SREBP-1c gene expression in the liver. Studies in transgenic mice show that 

constitutively active FoxO1 (CA-FoxO1) suppresses expression of SREBP-1c 

mRNA in liver by ∼50%, while SREBP-1c expression is increased ∼2-fold in 

liver-specifi c FoxO knockout mice. Similarly, studies in primary hepatocytes 

show that CA-FoxO1 suppresses SREBP1-c expression and inhibits basal and 

insulin-stimulated SREBP-1c promoter activity, while siRNA knockdown of 

FoxO1 increases SREBP-1c mRNA and protein levels and enhances SREBP- 

1c promoter activity. LXR, a nuclear receptor protein, plays an important 

role in promoting insulin-regulated SREBP-1c expression, and CA-FoxO1 

signifi cantly blunts the effects of LXR and LXR agonist TO901317 on the 

activity of the SREBP-1c promoter and a luciferase reporter construct driven 

by 3 LXR-response elements. Chromatin immunoprecipitation assays reveal 

that CA-FoxO1 reduces the recruitment of LXR to the SREBP-1c promoter 

despite the absence of FoxO-binding sites. Together, these studies indicate 

that FoxO proteins can suppress the expression of SREBP-1c and that this 

effect may be exerted, at least in part, by inhibiting recruitment of and 

transactivation by LXR. 

Supported by: Department of Veterans Affairs Merit Review Program 

& 1661-P 

Investigation of the Role of Irs1 Ser302 in Insulin Sensitivity/Resistance 

in Knock-In Mice 

KYLE D. COPPS, MORRIS F. WHITE, Boston, MA 

Hyperactivity of the nutrient-sensitive mTorc1→S6 kinase pathway impairs 

insulin-stimulated Akt activity in conjunction with serine phosphorylation 

and degradation of the insulin receptor substrates, Irs1 and Irs2. In particular, 

serine 307 of Irs1 (mouse S302) is a major site of phosphorylation by S6K 

in vitro, whereas phosphorylation of this site is decreased in tissue from 

insulin-sensitive S6k1 knockout mice. Thus, Ser302 phosphorylation might 

provide a link between abnormal physiologic states (potentially including 

nutrient excess) and insulin resistance. To test this hypothesis, knock-in mice 

were made having a non-phosphorylatable alanine (A) substitution at S302. 

At age 4 months, male S302A homozygotes (A/A) of mixed 129xC57BL/6 

background were signifi cantly glucose intolerant relative to similarly derived 

control knock-in mice (S/S) that expressed wild-type Irs1, but were also ∼10% 

more massive. By contrast, the A/A and S/S mice were indistinguishable 

by measurement of fasting blood glucose and insulin concentrations or 

sensitivity to IP-injected insulin. To more specifi cally assess the function of 

Irs, the A/A and S/S mice were intercrossed with mice bearing conditional 

(fl oxed) Irs1 and Irs2 alleles to generate males lacking Irs2 and retaining 

a single Irs1 A or S allele in the hepatocyte compartment. Comparison of 

these ‘genetically stressed’ mice (A/- or S/-) demonstrated the emergence of 

moderate glucose intolerance, but unaltered sensitivity to injected insulin, in 

A/- mice between 4-6 months of age. The A/- mice were also slightly larger 

on average than S/- mice at 6 months. Further experiments are in progress to 

carefully evaluate the signaling capacity of the mutant S302A Irs1 protein, in 

the context of both normal and abnormal liver physiology caused by mTorc1 

activation. 

& 1662-P 

Lecithin:Cholesterol Acyltransferase (LCAT) Defi cient Mice Are 

Resistant to Hepatic ER Stress and Insulin Resistance through 

Evasion of Cholesterol (Ch) Accumulation in the ER Membrane 

LAUREN S. HAGER, LIXIN LI, LU LIU, GRAHAM MAGUIRE, MARK NAPLES, CHRIS 

BAKER, LILIA MAGOMEDOVA, CAROLYN L. CUMMINS, PHILIP W. CONNELLY, 

KHOSROW ADELI, DOMINIC S. NG, Toronto, ON, Canada 

LCAT mediates the esterifi cation of free cholesterol (FC) in lipoproteins 

and its absence causes profound HDL defi ciency and hypertriglyceridemia. 

Although low HDL and high TG are typically seen in insulin resistance (IR), 

our lab has reported that chow-fed LCAT null mice in the LDL Receptor 

(LDLR) knockout background (DKO) are paradoxically more insulin sensitive 

than LDLR single knockout (SKO) controls. We observed that DKO mice are 



A452 

resistant to high fat high sucrose (HFHS) diet-induced obesity and IR. We 

also saw an increase in basal hepatic ER stress in SKO mice (2-fold increase 

in CHOP and BiP mRNA; p

TA correction can lead to loss of meaningful differences in GNG and GGL 

between study groups. Future studies are required to tease out differences 

in isolated fasting hyperglycemia IFG/NGT vs. IFG/IGT. 

Supported by: DK29953 

& 1664-P 

The Effect of Colesevelam Hydrochloride on Fasting and Postprandial 

Glucose Metabolism in Subjects with Type 2 Diabetes 

GALINA SMUSHKIN, CHIARA DALLA MAN, MATHENI SATHANANTHAN, PAULA 

D. GIESLER, CLAUDIO COBELLI, ADRIAN VELLA, Rochester, MN, Padua, Italy 

Lipid lowering therapy with a bile-acid sequestrant Colesevelam 

Hydrochloride in people with type 2 diabetes has been associated with 

a small but signifi cant decrease in HbA 1c. However the mechanism of this 

effect on glycemic control is unclear. To examine the effect of Colesevelam 

on fasting and postprandial glucose metabolism in type 2 diabetes we 

studied 40 subjects using a double-blind, placebo-controlled, parallel-group 

design. Subjects were studied at baseline and after a 12 week treatment 

period using a labeled mixed meal consisting of 50g of bacon, 2 scrambled 

eggs and 75g of Jell-O labeled with [1- 13 C]-glucose. [6- 3 H] glucose was 

infused intravenously to measure the systemic rate of meal appearance 

(Meal Ra). Infused [6,6- 2 H 2 ] glucose enabled measurement of endogenous 

glucose production (EGP) and glucose disappearance (Rd). Insulin sensitivity 

(Si) and β-cell responsivity indices (Φ) were estimated using the oral glucose 

and C-peptide minimal models, respectively. The Disposition Index (DI) for 

each individual was subsequently calculated. Therapy with Colesevelam 

was associated with a decrease in fasting (7.05±0.24 vs. 6.56±0.2mmol/l, 

p


Obesity 

POSTERS 

fasting state during elective cholecystectomy from lean subjects without 

DM2 (n=8) and during gastric bypass from obese subjects with or without 

DM2 (n=25). 

After adjusting for center, sex, and age by multiple linear regression 

analysis, α-HB was directly related to FLI (partial r = 0.21, p30% LFAT. Liver fat was associated with a reduction 

in plasma adiponectin and increased insulin resistance in adipose tissue 

(ATIR, %ins supprFFA) and liver (HIRI) (Table) with no further deterioration 

beyond LFAT 10%. There was no relationship between increasing BMI and 

LFAT (Table). Histologically, an increase in LFAT in NAFLD pts from 5-10% vs. 

any group LFAT>10% signifi cantly worsened liver infl ammation (p10%. In contrast, histology continues to worsen with LFAT up to 20-30%, 

but not beyond this point. 

No-LFAT 

(10-20%) 


Severe 

(>20-30%) 

Very Severe 

(>30%) 

Age (yrs) 60±3 59±2 54±2 55±1 54±1 

BMI (%) 33.1±2.1 35.5±1.6 34.8±1.3 35.0±1.1 35.0±1.1 

A1c (%) 6.6±0.4 6.7±0.3 7.3±0.2 7.2±0.2 6.8±0.2 

Adiponectin (µg/ml) 11.1±2.1 8.1±1.4 9.6±1.1 7.8±1.0 9.8±1.0 

ATIR (mmol/l · U/ml) 3±3 6±2 7±2 13±2*† 11±2*† 

% insulin supprFFA (%) 73±9 56±10 60±8 33±6*† 39±6*† 

HIRI(mg/kg-1 · min-1 · µU/ml) 5±2 27±9* 17±7* 26±5* 24±5* 

*p

with increased body weight and/or adiposity. However, other bloodborne 

and metabolic factors can infl uence SI and be more substantial 

predictors of SI under baseline, non-stimulated conditions. We performed 

a comprehensive assessment of metabolic function in healthy male dogs 

(total n=90) to identify key determinants of insulin sensitivity. Wholebody 

SI (SI clamp ), obtained from euglycemic clamp in all animals, was used 

as the primary outcome variable. SI was also measured independently by 

minimal model analysis of the IVGTT in a subset of animals (n=36). Total and 

regional (visceral, subcutaneous) adiposity were measured by MRI (n=90). 

Glucose-stimulated insulin response was measured either by stepwise 

hyperglycemic clamp or acute insulin response from the IVGTT (n=86 and 

36, respectively). Despite similar body weight (28.7±0.3 kg), baseline trunk 

adiposity varied nearly 8-fold (172 to 1363 cm 3 ), refl ecting a broad range 

of both visceral and subcutaneous fat mass. Variability was also refl ected 

in SI clamp (5.9 to 75.9 dl/min per kg per μU/ml). There was an expected 

negative association between fasting insulin and SI clamp (p


Obesity 

POSTERS 

& 1675-P 

Investigation of the Role of Hepatic Vagus Nerve on Hepatic Glucose 

Metabolism by Use of [6, 6- 2 H 2 ] Glucose 

TAZU TAHARA, KUNPEI TOKUYAMA, NORIKAZU OGIHARA, MASATOSHI KIKU- 

CHI, Tokyo, Japan, Ibaraki, Japan, Saitama, Japan 

It has been documented that portal glucose delivery creates an important 

signal to enhance liver glycogen deposition. The signaling pathway has been 

established as a “portal signal”. We have identifi ed that hepatic glycogen 

phosphorylase (GP) activity is a surrogate marker of portal signal, because 

GP activity was immediately reduced after glucose load from portal vein, but 

not from peripherally. To assess the role of hepatic vagus nerve on hepatic 

glucose metabolism, glucose was infused into unrestrained conscious rats 

via either portal (Po) or peripheral (Pe) vein at a constant rate, and parameters 

involved in hepatic glucose metabolism such as glucose production, glucose 

output and glycogen deposition and GP activity were measured. Rats were 

subjected to hepatic vagotomy (HV) or sham operation (SO) 7 days prior to 

the experiment. After 24-hour fasting, tracer amount of [6, 6- 2H 2] glucose 

was infused for 180 minute into the peripheral vein for equilibration, and 

kept infusion throughout the experiment. Then, glucose was infused portally 

or peripherally (13.5 mg/ kg /min) for 120 minutes. Metabolic parameters 

were determined by the mass isotopomer distribution analysis method. In 

SO, hepatic glucose output was signifi cantly reduced in Po than Pe after 30 

minutes glucose infusion (3.13 and 6.05 mg / kg / min), while the reduction 

in glucose output observed in SO was disappeared in HV, and kept higher 

levels (16.7 and 23.2 mg / kg / min in Po and Pe). Hepatic glycogen deposition 

was signifi cantly higher in Po than Pe after 120 minutes glucose infusion 

(57.1±2.1 vs 35.7±1.7μmol / g liver) in SO, whereas the glycogen deposition 

in Po was reduced in HV compared with SO during portal glucose infusion 

and was no siginifi cantly difference between Po and Pe (28.7±7.4vs 25.0±7.2 

μmol / g liver). GP activity in SO was lower in Po than Pe after 120 minutes 

glucose infusion(1.04±0.21 vs 1.54±0.03μmol / g liver), but the difference 

in GP activity between Po and Pe was not observed in HV (0.91±0.18 and 

0.93±0.17 μmol / g liver). From the data we conclude that hepatic vagus 

nerve is important for the regulation of hepatic glucose production and 

glycogen deposition. 

& 1676-P 

Control of Blood Glucose in the Absence of Hepatic Glucose Production 

Due to an Induction of Renal and Intestinal Gluconeogenesis 

by Gluca gon 

FABIENNE RAJAS, ELODIE MUTEL, AMANDINE GAUTIER-STEIN, AYA ABDUL- 

WAHED, MARTA AMIGO, ANNE STEFANUTTI, GILLES MITHIEUX, Lyon, France 

Since the pioneering work of Claude Bernard in the 19 th Century, the 

scientifi c community has considered the liver to be the major source of 

endogenous glucose production (EGP). We investigated the importance of the 

liver as a source of EGP, by using a mouse model of liver-specifi c deletion of 

the glucose-6 phosphatase catalytic subunit gene (LG6pc-/- mice), encoding 

an essential enzyme for glucose production. We previously showed that an 

absence of hepatic glucose release had no major effect on the fasting plasma 

glucose concentration. Instead, there was an early sequential induction of 

gluconeogenesis in the kidney and intestine, the alternative gluconeogenic 

organs. Here, we investigated molecular mechanisms underlying expression 

changes of gluconeogenic genes (Pck1 encoding phosphoenol carboxykinase 

and G6pc) in both kidney and intestine of LG6pc-/- mice. Male adult 

B6.g6pc lox/lox .SACre ERT2/+ mice were treated with tamoxifen to obtain 

Lg6pc-/-. Blood was withdrawn for metabolic studies from 6h-fasted mice. 

Chip assays were performed with phospho-CREB antibody on G6pc and Pck1 

promoters. At 6h of fasting, glucagon level was higher in LG6pc-/- mice than 

in control mice (538±9 pg/ml vs 280±19 pg/ml in control) while LG6pc-/- mice 

showed lower insulin levels than that of control mice. After confi rming the 

presence of glucagon receptors in the kidney and intestine, we showed that 

P-CREB bound to the G6pc promoter in both organs. In contrast, P-CREB was 

only bound to the Pck1 promoter in the intestine. Interestingly, we obtained 

identical results in fed control mice in response of an IP injection of glucagon. 

Complementary data showed that Pck1 expression was mainly induced in the 

kidneys by metabolic acidosis observed in LG6pc-/-. In conclusion, our study 

provides a defi nitive quantitative estimate of the capacity of extrahepatic 

gluconeogenesis to sustain fasting EGP, regardless of the contribution of the 

liver. Moreover, glucagon plays a central role to induce gluconeogenesis in 

the kidney and intestine, as it is well known in the liver. 

Supported by: Agence Nationale de la Recherche and Association Française des 

glycogénoses 



A456 

& 1677-P 

Attenuation of Hepatic Steatosis Is Associated with Increased 

Hepatic Phospholipid Eicosapentaenoic Acid (EPA) and Docosahexaenoic 

Acid (DHA) in Diet-Induced Obese (DIO) Rats Fed a Canola/ 

Flax Oil Blend 

DANIELLE P. HANKE, CARLA G. TAYLOR, PETER C. ZAHRADKA, Winnipeg, MB, 

Canada 

Insulin resistance is the most important pathogenic factor in the etiology 

of non-alcoholic fatty liver disease (NAFLD). NAFLD has been associated with 

reduced omega-3 (n-3) fatty acid status. It has been proposed that n-3 fatty 

acids may prevent hepatic steatosis and progression to NAFLD by increasing 

hepatic fatty acid oxidation via peroxisome proliferator activated receptor 

a (PPARa) and decreasing hepatic fatty acid synthesis via sterol regulatory 

element-binding protein-1c (SREBP-1c). Furthermore, the effi cacy of the plantbased 

n-3 fatty acid, α-linolenic acid (ALA), as a dietary precursor of EPA and 

DHA for modulating hepatic steatosis is unknown. Thus, the present study 

investigated hepatic steatosis, hepatic fatty acid composition and markers 

of fatty acid oxidation and synthesis in 6 week old Obese Prone Sprague 

Dawley rats fed high fat (55% energy) diets containing various fat types 

including high oleic canola oil, canola oil, a canola/fl ax oil blend (C/F, 3:1), 

saffl ower oil, soybean oil, or lard for 12 weeks. Upon completion of the study, 

the C/F and weight matched (WM) groups had the lowest percent liver lipid. 

The C/F group had the highest amount of ALA in their diet and this resulted 

in the highest total n-3 and EPA in hepatic PL. The C/F group also had one of 

the highest DHA and lowest arachidonic acid (n-6) concentrations in hepatic 

PL. Conversely, the type of fat or oil did not affect hepatic protein levels 

of PPARa and acyl-CoA oxidase or hepatic protein levels of SREBP-1c and 

acetyl-CoA carboxylase. In conclusion, our data indicates that the C/F diet, 

which was high in the plant-based n-3 ALA, can attenuate hepatic steatosis 

and favourably alter hepatic fatty acid concentrations of PL by increasing 

EPA and DHA. However, the reduction in hepatic steatosis by the canola/fl ax 

oil blend was not explained by the upregulation of PPARa and/or inhibition or 

suppression of SREBP-1c. 

Supported by: Canola Council of Canada 

& 1678-P 

Overexpression of SIRT1 in the Liver Deacetylates X-Box Binding 

Protein 1 and Attenuates Hepatic Steatosis and Insulin Resistance 

in ob/ob Mice 

YU LI, KAZUTO NAKAMURA, WALSH KENNETH, MENGWEI ZANG, Boston, MA 

Endoplasmic reticulum (ER) stress has been implicated in the pathophysiology 

of human type 2 diabetes (T2DM). Small molecular activators of SIRT1 were 

shown to prevent diet-induced obesity and insulin resistance. Our previous 

studies have indicated that SIRT1 and resveratrol regulate hepatocyte lipid 

metabolism through activating AMPK. To extend these fi ndings, the goal of 

the present study was to explore the mechanism of the therapeutic impact 

of NAD-dependent deacetylase SIRT1 on metabolic deterioration in T2DM. 

We demonstrated that adenovirus-mediated overexpression of SIRT1 in 

the liver of ob/ob mice attenuated hepatic steatosis, lowered hepatic 

lipids contents, and ameliorated glucose tolerance and systemic insulin 

resistance. Enhanced insulin sensitivity by SIRT1 was associated with 

decreased ER stress markers such as GRP78 and CHOP. Importantly, X-box 

binding protein 1(XBP1), a critical ER stress marker, was identifi ed as a direct 

target of SIRT1, since co-immunoprecipitation showed the spliced form of 

endogenous XBP1 physically interacted with SIRT1 in the mouse embryonic 

fi broblasts (MEFs). Hepatic overexpression of SIRT1 increased its interaction 

with spliced XBP1 and resulted in the deacetylation of spliced XBP1 in ob/ 

ob mouse livers, as compared to those of Ad-GFP-injected mice. Hepatic 

overexpression of SIRT1 also decreased SREBP-1c, key enzyme involved 

in lipid synthesis, in ob/ob mouse livers. In contrast, the promoter activity 

of SREBP-1c target, fatty acid synthase, was increased by SIRT1 -/- MEFs, 

compared with SIRT1 +/+ MEFs. Interestingly, serum levels of infl ammatory 

marker monocyte chemotactic protein-1 (MCP-1) were reduced upon SIRT1 

treatment. Moreover, resveratrol decreased tunicamycin-induced NF-κB 

promoter activity and mRNA levels of proinfl ammatory cytokine TNFα in 

HepG2 cells. Taken together, our results 1) identify XBP1 as a novel target of 

SIRT1 through the deacetylation regulation in the liver and in vitro; 2) reveal 

that deacetylation of XBP1 by SIRT1 in the liver may serve as a therapeutic 

strategy to attenuate obesity-induced ER stress, infl ammation, hepatic 

steatosis, and insulin resistance. 

& Guided Audio Tour poster ADA-Funded Research

& 1679-P 

A-769662-Mediated AMPK Activation in the Liver Reverses Hepatic 

Steatosis in Lipodystrophic Mice 

MARC FORETZ, JOCELYNE LECLERC, BENOIT VIOLLET, Paris, France 

AMP-activated kinase (AMPK) is a key regulator of lipid metabolism 

which increases fatty acid oxidation and decreases lipid synthesis. Leptin 

and adiponectin, two adipokines secreted by adipose tissue, activate AMPK 

and stimulate lipid oxidation. Lipodystrophic syndrome is characterized by 

loss of adipose tissue depots associated with low adipokine levels, hepatic 

steatosis and severe insulin resistance. We hypothesized that fatty liver in 

lipodystrophy may be in part explained by an alteration in AMPK activity. 

Inversely, pharmacological activation of AMPK in the liver may alleviate 

hepatic steatosis associated with lipodystrophy. 

We measured AMPK activity in liver from aP2-SREBP-1c mice, a 

lipodystrophy model transgenic mice. We found that AMPK activity was 

decreased by 50% and phosphorylation of acetyl-CoA carboxylase (ACC), 

its downstream substrate, was reduced by 40%. Moreover, hepatic fatty 

acid oxidation assessed by the plasma concentration of ketones bodies was 

decreased by 50% and hepatic triglyceride (TG) content was increased by 

5-fold. The small molecule A-769662 has been identifi ed as a potent and 

direct activator for AMPK. Treatment of aP2-SREBP-1c mice with A-769662 

for 1 week at a dose of 30 mg/kg b.i.d. increased phosphorylation of AMPK 

and phospho-Ser79-ACC/ACC ratio in liver. Hepatic fatty acid oxidation was 

completely restored. Importantly, hepatic TG and cholesterol content were 

decreased by 10-fold and 20%, respectively. 

In conclusion, AMPK activity and fatty acid oxidation were decreased 

in the liver of lipodystrophic mice whereas A-769662 treatment led to 

increased rates of fatty acid oxidation and abolished fatty liver. Our study 

suggests that small molecule AMPK activators may have therapeutic 

potential to reverse hepatic steatosis in patients with lipodystrophy or other 

metabolic diseases. 

1680-P 

De Novo Lipogenesis Induced Hepatic Steatosis and Insulin Resistance 

Involve ER Stress but Not Infl ammation in Contrast to Lipid 

Oversupply 

LU-PING REN, STANLEY MH CHAN, XIAO-YI ZENG, ROSS D. LAYBUTT, TRISTAN 

J. ISELI, EDWARD W. KRAEGEN, GREGORY J. COONEY, NIGEL TURNER, JI-MING 

YE, Sydney, Australia, Melbourne, Australia 

Mitochondrial dysfunction, infl ammation and endoplasmic reticulum (ER) 

stress have been implicated in hepatic steatosis and insulin resistance in 

genetically obese and insulin resistant mice. The present study investigated 

the role of mitochondrial dysfunction, infl ammation and ER stress in the 

development of hepatic steatosis and insulin resistance due to hepatic 

de novo lipogenesis from a high fructose diet (HFru), compared to that of 

extrahepatic lipid oversupply from a high fat diet (HFat). Mice fed HFru or 

HFat developed marked hepatic steatosis (triglyceride increased by 3.5 and 

2.4 fold, p


Obesity 

POSTERS 

novel long-term model indicates that diabetes contributes to progression 

from NASH to more end-stage liver disease. Subsequent studies will further 

defi ne molecular mechanisms of this process. Supported by NHMRC of 

Australia Project Grant #632822. 

Supported by: NHMRC of Australia Project Grant No. 632822 

1683-P 

Differential Response of Mitochondrial Enzymes to Obesity and 

Excessive Dietary Fat Intake 

ELIZAVETA V. MENCHIKOVA, WAN HUANG, FREDERICO G.S. TOLEDO, ROBERT 

M. O’DOHERTY, BRET H. GOODPASTER, VLADIMIR B. RITOV, Pittsburgh, PA 

The current study was undertaken to compare the effect of obesity or 

excessive fat intake on specifi c activity (per mitochondria mass) of electrontransport 

chain (NADH-oxidase) and TCA cycle (citrate synthase (CS) that 

provides entrance of acetyls into TCA cycle) enzymes in human skeletal muscle 

and rat liver. Tissue cardiolipin (CL) content was measured as independent 

non-enzymatic marker of mitochondrial mass. We compared CL, NADHoxidase 

and CS activity in skeletal muscle biopsies from sedentary lean 

insulin sensitive (L, n=9, age 47±2, BMI 24±1) and sedentary obese insulin 

resistant (Ob, n=15, age 42±3, BMI 35±1) subjects. There is no difference 

in CL content in skeletal muscle from L and Ob (74±9 and 76±6 μg/mU CK, 

respectively). The activity of NADH-oxidase was reduced signifi cantly in Ob 

in comparison with L (5.63±1.59 and 2.45±0.23 U/mg CL (P=0.02) for L and Ob 

subjects, respectively). However, the citrate synthase activity is signifi cantly 

higher in obesity in comparison with lean group (35.8±3.4 and 54.5±5.3 U/mg 

CL (P=0.02) for L and Ob, respectively). 

Analysis of liver mitochondria was performed on a rodent high-fat feeding 

model. Five weeks of high–fat feeding in rats (n=4) induces insulin resistance, 

decreases specifi c NADH-oxidase activity (1.470±0.53 and 0.997±0.08 U/ 

mg CL for control (C) and high fat-fed (HF) rats, respectively), signifi cantly 

increases citrate synthase activity (19.7±2.1 and 27.4±1.4 U/mg CL (P=0.023) 

for C and HF rats, respectively) and slightly decreases liver cardiolipin 

content. We hypothesize that excessive dietary fat intake induces activation 

of CS (by induction of protein synthesis or by post-translational modifi cation 

in liver and skeletal muscle) to utilize excessive acetyl-CoA produced in 

beta-oxidation pathway. 

These data highlight that mitochondrial TCA cycle and electron transport 

chain enzymes respond differently to energy excess. Further investigation is 

needed to better understand the respective roles of specifi c mitochondrial 

function in obesity and insulin resistance. ADA-Funded Research 

1684-P 

Effect of Ecdysterone on Hepatic AMP Activated Protein Kinase in 

Mice Fed with a High-Fat Diet 

ZHONG Q. WANG, YONGMEI YU, XIAN H. ZHANG, DAVID RIBNICKY, WILLIAM T. 

CEFALU, Baton Rouge, LA, New Brunswick, NJ 

Chronic nutrient overload leads to obesity and metabolic disorders, 

including insulin resistance and type 2 diabetes. Ecdysterone (Ecdy), 

specifi cally20-hydroxyecdysone, is a steroid hormone from plants). Our 

previous study showed that Ecdy may have anti-obesity and anti-diabetic 

effects, but its molecular mechanisms remain largely unknown. It is well 

documented AMP activated protein kinase (AMPK) plays a key role in glucose 

metabolism. In order to evaluate the effect of Ecdy on AMPK signaling, we 

examined hepatic AMPK pathway proteins in mice fed a low-fat diet (control) 

or high-fat diet (HFD) with or without low dose (25 mg/kg body weight, Ecdy 

L) or high-dose (50 mg/kg, Ecdy H) of Ecdy for 12 weeks. Body weight, fasting 

glucose and insulin concentrations were measured as well. At study end, 

no differences in body weight, food intake or energy expenditure were 

observed between HFD groups with or without Ecdy L, but the body weight 

was signifi cantly lower in Ecdy H group relative to the HFD group (P

1687-P 

Free Fatty Acid-Induced Hepatic Insulin Resistance Is Mediated by 

Endoplasmic Reticulum Stress 

CRISTINA DIRLEA, SANDRA PEREIRA, ADRIA GIACCA, Toronto, ON, Canada 

Insulin resistance is a common feature of obesity and is primarily caused 

by an increased release of free fatty acids (FFA) and altered release of 

adipokines from the expanded adipose tissue. FFA-induced insulin resistance 

has different mechanisms depending on the tissue type and although 

previous work in our lab has shown activation of c-Jun-N-terminal kinase 

(JNK) in the liver upon short-term lipid infusion, whether JNK is causal in 

FFA-induced insulin resistance is unknown. One potential mechanism of 

JNK activation is as a result of endoplasmic reticulum (ER) stress. Therefore, 

the present study investigates the causal roles of ER stress and JNK in 

FFA-induced insulin resistance. Wistar rats were infused intravenously 

for 7 hours with saline or Intralipid plus heparin to elevate plasma FFA 

with or without an ER stress inhibitor (4-phenylbutyrate (4-PBA)) or a JNK 

inhibitor (SP600125). Insulin-induced suppression of endogenous glucose 

production during hyperinsulinemic-euglycemic clamp was measured using 

tracer methods. 4-PBA co-infusion prevented FFA-induced hepatic insulin 

resistance (P


Obesity 

POSTERS 

In this study, we identify the histone demethylase plant homeodomain 

(PHD) fi nger 2 (phf2) as an important regulator of both glycolytic and lipogenic 

gene expression acting as an upstream regulator of ChREBP activity. In 

cultured mouse hepatocytes, using chromatin immunoprecipitation (ChIP) 

analysis, we show that phf2 is recruited to the ChoRE containing region 

of both glycolytic and lipogenic gene promoters (including LPK and FAS) 

in response to glucose and insulin stimulation. Then, phf2 specifi cally 

demethylates di-methylated lysine 9 on histone H3 (H3K9Me2) allowing the 

recruitment of ChREBP to these promoters to increase fatty acid synthesis. 

In the opposite, phf2 silencing results in increased H3K9Me2 methylation at 

these promoters, resulting in decreased of ChREBP occupancy and inhibition 

of fatty acid synthesis. Finally, in liver of fed diabetic db/db mice, high phf2 

activity at these glycolytic and lipogenic gene promoters is associated with 

decreased H3K9Me2 methylation and increased ChREBP transcriptional 

activity, correlating with the development of hepatic steatosis. Our fi ndings 

suggest that inhibition of phf2 activity in liver may be benefi cial for treating 

hepatic steatosis in state of obesity and type 2 diabetes. 

Supported by: Agence Nationale de la recherhe (ANR-09-JCJC-0057-01) and by 

the FRM 

1692-P 

Prenatal Protein Defi ciency Alters Circadian Rhythms in Core Clock 

Oscillator Protein Expression in the Offspring 

ARMAND V. CENTANNI, DIANA C. ALBARADO, GREGORY M. SUTTON, Baton 

Rouge, LA 

The mechanisms linking intrauterine growth retardation (IUGR) with 

adulthood obesity and diabetes are unknown. Previous studies performed 

in our lab have demonstrated an altered circadian phenotype in 8 wk old 

male C57Bl/6J mice subjected to protein malnutrition (undernourished 

offspring, UO) in utero coupled with altered glucose homeostasis. Gene 

expression of the nuclear receptor, Rev-erbα, a component of the circadian 

clock mechanism in liver was dramatically reduced and out of phase in UO at 

8 wk of age compared to control offspring (CO). Rev-erbα repressed genes 

involved in circadian regulation (Bmal1 and Per2) were increased in UO mice. 

Surprisingly, protein expression of Rev-erbα in UO liver did not oscillate and 

was expressed at similar levels at circadian time points (CT) 1200 and 2400 

(noon and midnight) suggesting gene expression and protein translation are 

misaligned compared to CO. Phosphorylation of glycogen synthase kinase- 

3β and protein kinase B, two second messenger signaling molecules that 

play a role both directly (GSK3β) and indirectly (PKB) in Rev-erbα regulation. 

We now demonstrate that phosphorylation at the GSKβ site on Rev-erbα 

is dramatically reduced in UO liver, suggesting altered transcription and 

regulation. These data suggest potentially cell surface signaling, perhaps 

through insulin may play a role in impaired clock controlled processes in 

UO mice. We conclude that UO mice exhibit a metabolic disorder involving 

abnormal circadian patterns of gene and protein expression. Altered Reverbα 

expression and function may be a key factor in metabolic dysregulation 

associated with IUGR. ADA-Funded Research 

1693-P 

Pyruvate Carboxylase, a Novel Therapeutic Target for Type 2 Diabetes 

NAOKI KUMASHIRO, SARA A. BEDDOW, IOANA FAT, SACHIN K. MAJUMDAR, 

FITSUM GUEBRE-EGZIABHER, JENNIFER L. CHRISTIANSON, PRASAD MANCHEM, 

BRETT P. MONIA, SANJAY BHANOT, GERALD I. SHULMAN, VARMAN T. SAMUEL, 

New Haven, CT, Carlsbad, CA 

Fasting hyperglycemia in T2D is due to increased gluconeogenesis, a 

process for which the enzymatic regulation is poorly understood. We recently 

observed an association between fasting hyperglycemia and expression 

of pyruvate carboxylase (PC) but not PEPCK and G6Pase in several rodent 

models and thus, hypothesized that PC is an excellent therapeutic target for 

T2D. To test this, we used antisense oligonucleotides (ASO’s) to decrease 

PC expression in normal SD rats fed a regular chow, high-fat fed (HFF) SD 

rats and Zucker Diabetic Fatty (ZDF) rats. ASO’s specifi cally decrease target 

expression in liver and adipose but not other tissues (e.g. pancreas). PC ASO 

treatment decreased PC expression by ∼90%. In regular chow fed rats, this 

reduced plasma glucose concentrations in both the fasted [Cont: 109±1 vs. 

PC: 103±1, (P

for a period of 3-4 months (n=7 per group; p


Obesity 

POSTERS 


INTEGRATED PHYSIOLOGY—MACRONUTRIENT CATEGORY METABOLISM AND FOOD INTAKE 

In vivo model, Ad-Clu in liver of mice by tail vein injection inhibited 

hepatic steatosis through inhibition of SREBP-1c expression. Additionally, 

Clusterin inhibited LXR-agonist-stimulated LXR-DNA binding to its response 

consensus element on SREBP-1c promoter. This study shows that Clusterin 

inhibits hepatic lipogenesis in vitro and in vivo through downregulation of 

SREBP1c. The present study suggested that clusterin plays a critical role in 

the regulating hepatic lipogenesis. 

INTEGRATED PHYSIOLOGY—MACRONUTRIENT 

METABOLISM AND FOOD INTAKE 

[See also: Presidents Poster 455-PP, page A126.] 

Guided Audio Tour: Macronutrients and Metabolism (Posters 1700-P to 

1708-P), see page 13. 

& 1700-P 

Infl uence of Age on the Brain Response to Eating in Adults Evaluated 

Using Continuous Arterial Spin Labeling Functional Magnetic Resonance 

Imaging 

SARAH LEE, YEE S. CHEAH, YASHICA NATHAN, BULA WILSON, ANDREW 

PERNET, MICHAEL J. BRAMMER, STEPHANIE A. AMIEL, FERNANDO O. ZELAYA, 

London, United Kingdom 

Appetite and satiety dysregulation may result in overeating, obesity and 

Type 2 diabetes. Central control of appetite is an attractive target for weight 

reduction and insulin sensitising therapies. Functional magnetic resonance 

imaging (fMRI) using continuous arterial spin labeling (cASL), a non-invasive 

method for quantifying regional cerebral blood fl ow (rCBF), has been used to 

investigate the responses to a mixed meal ingestion, compared to water, in 

2 groups of healthy volunteers with a signifi cant age difference. 

11 healthy young adults (YA, 4 female): aged 19-33 yrs and 11 older adults 

(OA, 8 female) aged 38-52 yrs (p


& 1703-P 

Meal-Induced Insulin Secretion Is Associated with Postprandial 

Triglyceride Clearance in Subjects Consuming Glucose, Fructose or 

High Fructose Corn Syrup-Sweetened Beverages 

ROEL G. VINK, KIMBER L. STANHOPE, ANDREW A. BREMER, VALENTINA MEDICI, 

GUOXIA CHEN, TAK HOU FONG, VIVIEN LEE, ROSE MENORCA, NANCY L. KEIM, 

PETER J. HAVEL, Davis, CA, Nashville, TN 

It has been proposed that the adverse metabolic effects of chronic 

consumption of sugar-sweetened beverages are a consequence of increased 

meal-induced glucose and insulin excursions, i.e. dietary glycemic index. 

Our objective was to investigate the role of post-meal insulin peaks, which 

activate lipoprotein lipase (LPL), in postprandial triglyceride (TG) clearance in 

48 adults (age: 18-40 years, BMI: 18-35 kg/m²) consuming glucose, fructose 

or high fructose corn syrup (HFCS) sweetened beverages. For 12 days 

subjects resided at home and consumed their usual ad libitum diet along 

with glucose, fructose or HFCS (16/group) sweetened beverages at 25% of 

total energy requirements. Subsequently, fasting ApoC3 and 23-h insulin and 

TG levels were measured at the clinical research center while the subjects 

consumed the sweetened beverages with meals with an energy balanced 

diet (25% sugar-sweetened beverage, 30% complex carbohydrate, 30% 

fat, 15% protein). Net TG clearance was calculated as the decrease of TG 

2 hours after the post-dinner TG peak. Consumption of glucose-sweetened 

beverages resulted in the largest glucose and insulin peaks and 23-h AUC. 

These levels were lowest in subjects consuming fructose and intermediate 

in subjects consuming HFCS (effect of sugar P


Obesity 

POSTERS 



& 1707-P 

Inhibition of Delta-6 Desaturase Increases Phospholipid Linoleate 

Content, Improves Cardiac Mitochondrial Function and Restores 

Glucose Tolerance in ob Mice 

MELISSA A. ROUTH, CHRISTOPHER M. MULLIGAN, CATHERINE H. LE, JULIANO 

SILVEIRA, GERRIT T. BOUMA, GENEVIEVE C. SPARAGNA, SIMONA ZARINI, 

ROBERT C. MURPHY, ADAM J. CHICCO, Fort Collins, CO, Boulder, CO, Aurora, CO 

Epidemiological studies have linked a lower proportion of linoleic acid (LA, 

18:2n6) in serum phospholipids (PLs) to insulin resistance and cardiovascular 

mortality in humans, but the mechanism and pathophysiological basis of 

this phenomenon are unclear. A selective loss of LA also occurs in PLs from 

multiple tissues in obese/insulin-resistant (ob) vs. lean (C57Bl/6) mice, which 

is paralleled by increases in long-chain PUFAs and an accumulation of their 

eicosanoid derivatives. We hypothesized that this PL remodeling results 

from increased activity of delta-6 desaturase (D6D), the rate limiting enzyme 

in long-chain PUFA biosynthesis that utilizes LA as a substrate, and explored 

potential pathophysiological consequences of this phenomenon in ob mice. 

qRT-PCR revealed a 1-2 fold induction of D6D mRNA in ob vs. lean mice 

that paralleled increases in PL PUFA molar% ratios indicative of greater D6D 

activity in heart, liver, serum and muscle (P < 0.01). Treatment of 4 mo old 

male ob mice with the selective D6D inhibitor SC-26196 (SC, 100 mg/kg/d 

for 4 wks) reversed D6D indices and increased the molar% of LA in tissue 

PLs, and ameliorated 2 to 20-fold elevations in several pro-infl ammatory 

eicosanoid species present in the serum and hearts of untreated ob mice. SC 

treatment completely normalized the fatty acid composition of cardiolipin in 

cardiac mitochondria, which was associated with improvements in the rate 

(state 3) and effi ciency (ADP/O ratio) of mitochondrial respiration (P < 0.05). 

Interestingly, D6D inhibition also improved response to an acute glucose 

challenge in ob mice (1 mg/g BW i.p.), indicated by lower peak (146% vs. 

219% of fasting) and fi nal (85 vs. 180% fasting 2 hrs-post challenge) blood 

glucose levels vs. untreated ob mice (P < 0.01). These studies indicate that 

D6D plays a pivotal role in PL remodeling and eicosanoid production in ob 

mice, and highlight the need for further investigation of PUFA metabolism 

in the development of glucose intolerance and cardiac mitochondrial 

dysfunction associated with obesity/insulin resistance. 

Supported by: AHA Grant 

& 1708-P 

Improving Mitochondrial Fat Oxidation by Restoring Tissue Glutathione 

Concentrations Increases Insulin Sensitivity, and Lowers 

Hepatic Fat, Triglycerides and Body Weight in Aged Mice 

RAJAGOPAL V. SEKHAR, DAN NGUYEN, SUSAN L. SAMSON, VASUMATHI T. 

REDDY, Houston, TX 

Introduction: We tested the hypotheses that glutathione defi ciency underlies 

impaired mitochondrial fat oxidation in aging, and contributes to insulin resistance, 

elevated hepatic fat and obesity, and whether improving tissue glutathione 

concentrations with dietary precursors would restore these defects. 

Methods: 8 young (18w) and 16 old (76w) male C57BL/6 mice were 

studied. The old mice were matched for age, sex, weight, glucose tolerance 

and fat oxidation: one group (‘treated’) was pair fed to the other (‘untreated’) 

control group using an isocaloric-isonitrogenous diet for 6w (both groups 

ate identical daily calories and protein nitrogen - diet of the treated group 

had higher content of glycine and cysteine (as n-acetylcysteine), precursor 

amino-acids for glutathione). 

Outcome measures were liver and muscle glutathione, body weights 

and composition, fat oxidation, glucose and insulin tolerance, plasma 

triglycerides, liver and renal profi les, and hepatic fat content. 

Results: Compared to young mice, untreated old mice had30-35% lower 

glutathione levels in the liver and muscle, 20% lower mitochondrial fattyacid 

oxidation, and signifi cantly higher hepatic fat, body weight, total body 

fat, and plasma triglycerides. 

Compared to the untreated old control mice, the treated mice showed 

signifi cantly improved liver and muscle glutathione concentrations by 20- 

36%, and mitochondrial fatty acid oxidation by 22%, completely restored 

liver fat content, and improved body weight by 16%, triglycerides by 36%, and 

insulin sensitivity by 66%, and no changes in expression of glutamylcystinyl 

ligase, an enzyme regulating glutathione synthesis. 

Conclusions: Glutathione defi ciency contributes to impaired mitochondrial fattyacid 

oxidation in aging, and predisposes to elevated hepatic fat, insulin resistance, 

dyslipidemia and obesity. Restoring glutathione concentrations reverses these 

defects. Using simple, safe and inexpensive amino-acid supplementation to 

boost glutathione levels may be developed as a novel nutritional approach to 

treat fatty liver disease, obesity and insulin resistance in aging. 

Supported by: Baylor Seed Fund 

A464 

1709-P 

Amino Acid Metabolites and Insulin Resistance in Human 

BRIAN A. IRVING, AUDREY J. WEYMILLER, HUSNAIN SYED, HELEN KARAKELIDES, 

MATTIAS SOOP, RICKEY E. CARTER, K. SREEKUMARAN NAIR, Rochester, MN 

Elevations in amino acids (AA), particularly branched chain AA (BCAA), 

have been reported to contribute to the development of insulin-resistance. 

We sought to determine the additive effects of AA, AA metabolites, and 

body composition on insulin sensitivity (SI). 24 non-diabetic lean (49+5 y, 

22.8+0.4 kg/m2 ), 27 non-diabetic obese (49+4 y, 30.4+0.4 kg/m2 ), and 

12 type 2 diabetic obese (57+4 y, 30.2+1.0 kg/m2 ) adults were studied. SI 

was assessed using the steady-state glucose infusion rate (GIR, umol/ 

kgFFM/min) obtained during a standardized hyperinsulinemic (1.5 mU/ 

kgFFM/min) euglycemic clamp. Body composition was determined using 

DEXA. Ultra-performance LC-MS/MS was used to quantify fasting AA 

and AA metabolites. Compared to the non-diabetic lean GIR (53+3) was 

progressively lower in both non-diabetic obese (40+3, p

1711-P 

The CB1 Inverse Agonist Rimonabant Prevents Obesity in Mice by 

Increasing Energy Expenditure Rather Than by Reducing Food Intake 

KATY JANE BROWN, HONG WANG, JENNIFER H. YOON, ROBERT H. ECKEL, Denver, 

CO 

The cannabinoid receptor CB1 in the CNS plays an important role in 

regulating energy balance while peripheral CB1 receptors also appear to 

modulate nutrient metabolism. Previously, treatment of ob/ob mice and dietinduced 

obese mice with the CB1 receptor inverse agonist rimonabant (RM) 

caused decreased food intake, weight loss, and increased muscle glucose 

uptake and oxygen consumption. We now tested the effect of RM on 

preventing diet-induced obesity in C57BL/6J mice. At 8-wks male mice were 

ad libitum fed with either a 45% high fat (HF) diet or a 10% fat diet (HC), and 

also treated with RM (10 mg/kg/day) or saline by gastric gavage. Weight and 

food intake were monitored daily and an IP glucose tolerance test (GTT) and 

indirect calorimetry was completed after 7 wks of feeding and treatment. 

Our data show that RM prevented weight gain of mice on both HF and HC 

diets, making RM mice on both diets weigh similarly and less (10%↓, p


Obesity 

POSTERS 

prevented the THAP- and TUNI-induced decreases in insulin-stimulated 

IRS1 Y 612 phosphorylation and glucose uptake. These data suggest a novel 

paradigm where ER stress enhances TRB3 expression leading to insulin 

resistance in skeletal muscle. ADA-Funded Research 

& 1715-P 

Effects of Muscle Specifi c Deletion of Carnitine Palmitoyltransferase 

and Carnitine Acetyltransferase on Fuel Selection 

JINGYING ZHANG, ROBERT C. NOLAN, SARAH E. SEILER, TIMOTHY R. KOVES, 

INDU KHETERPAL, ROBERT D. STEVENS, OLGA R. ILKAYEVA, DEBORAH M. 

MUOIO, RANDALL L. MYNATT, Baton Rouge, LA, Durham, NC 

Mitochondrial fatty acid import and oxidation is initiated by carnitine 

palmitoyltransferase-I (CPTI). CPTI is located in the outer mitochondrial 

membrane and catalyzes the formation of long chain acyl-carnitines from 

carnitine and acyl-CoA. Short chain acyl-carnitines are formed via the 

activity of carnitine acetyltransferase (CrAT) in the mitochondrial matrix. 

Even though CrAT’s role is not completely understood, it is thought to buffer 

acetyl-CoA/CoA in the mitochondria. To understand the role of each enzyme 

in energy homeostasis, substrate utilization and insulin sensitivity; mice 

were generated with muscle specifi c deletion of CPT-I m-/- and CrAT m-/- . CPT- 

I m-/- and CrAT m-/- mice have normal body weight, fat mass, fat free mass, 

energy expenditure and food intake. However, long chain fatty acid oxidation 

is greatly reduced in muscle homogenates and isolated mitochondria 

from CPT-I m-/- mice. Given the popular hypothesis that impaired fatty acid 

oxidation in skeletal muscle causes the accumulation of lipid intermediates 

leading to insulin resistance, we paradoxically observed improved GTT 

and ITT in CPT-I m-/- mice relative to control mice. Metabolic chamber data 

demonstrated that CPT-I m-/- mice oxidize more carbohydrate than control 

mice. Conversely, CrAT m-/- mice had impaired GTT and ITT relative to control 

mice. Metabolic chamber data revealed no difference in 24 hour RER 

between CrAT m-/- and control mice, but there is impaired switching from 

fatty acid oxidation to carbohydrate oxidation during the transition from a 

fasted to fed state. In vitro studies in muscle homogenates and isolated 

mitochondria demonstrated that the addition of carnitine increased PDH 

activity and the complete oxidation of pyruvate in control mice but not CrAT 

m-/- mice. In addition, the ability of pyruvate to suppress fatty acid oxidation 

was blunted in mitochondria from CrAT m-/- mice. Together our data indicate 

that, the loss of skeletal muscle CPTI results in reduced oxidization of long 

chain fatty acids and stimulates glucose oxidation, whereas the loss of CrAT 

in muscle predominately affects metabolic fl exibility. 

Supported by: NIH ADA-Funded Research 

& 1716-P 

Diabetes Mellitus Impact on Left Ventricular Myocardial Structure 

and Function in Aortic Stenosis before Valve Replacement 

INÊS FALCÃO-PIRES, NAZHA HAMDANI, CRISTINA GAVINA, JOLANDA VAN DER 

VELDEN, ATTILA BORBÉLY, HANS NIESSEN, GER STIENEN, ADELINO F. LEITE- 

MOREIRA, WALTER J. PAULUS, Porto, Portugal, Amsterdam, The Netherlands, 

Debrecen, Hungary, Amsterdam, Portugal 

Diabetes mellitus (DM) is an independent risk factor for progression of 

aortic valve stenosis (AS) and signifi cantly impacts longterm outcome after 

valve replacement. High incidence of residual heart failure may account 

for this prognosis. We aimed to assess the impact of DM on diastolic (dys) 

function of AS patients. 

Patients with severe isolated AS (n=46) and AS plus type-II diabetes 

(AS-DM + ,n=16) with preserved left ventricular (LV) ejection fraction and 

no clinical or angiographic signs of coronary artery disease were studied. 

Doppler echocardiographic data was used to compare in vivo LV function. 

Biopsies were used to assess fi brosis, cardiomyocyte hypertrophy 

(MyD), advanced glycation endproducts (AGEs) and phosphorylation of 

myofi lamentary proteins. Isolated and permeabilized cardiomyocytes were 

used to measure active force (F active ), resting force (F passive ) and myofi lament 

calcium sensitivity(pCa 50 ). 

In isolated AS, LV deceleration time and end-diastolic pressure were 

augmented (239±19ms; 21.4±1.4mmHg, respectively) and the latter 

signifi cantly correlated with increased fi brosis (12.9±1.1%; r=0.40,p=0.04) 

and MyD (22.9±0.5μm; r=0.60, p

the curve analysis; LF-C: 2534±124, HF-C: 3371±186 mM*min, P


Obesity 

POSTERS 

inhibitory protein Bcl2. Phospho proteomic analysis and phospho-specifi c 

immunoblotting demonstrated an increased tyrosine phosphorylation of 

STAT3, a known transcriptional activator of Bcl2 expression. Expression of 

a dominant-interfering STAT3 mutant (STAT3-Y705A) was a potent activator 

of skeletal muscle macroautophagy. Together these data demonstrate that 

Fyn functions as a suppressor of macroautophagy through a STAT3 mediated 

inhibition of the Vps34/Beclin1/ATG14 complex 1. 

Supported by: NIH 

& 1723-P 

Macrophages Are Necessary for Exercise-Induced Enhancement of 

Insulin Sensitivity in Skeletal Muscle 

SHIN-ICHI IKEDA, YOSHIFUMI TAMURA, SAORI KAKEHI, YOSHIO FUJITANI, 

TAKAHISA HIROSE, RYUZO KAWAMORI, HIROTAKA WATADA, Tokyo, Japan 

Type 2 diabetes and obesity are characterized by insulin resistance 

in skeletal muscle. It has been well demonstrated that exercise increase 

insulin sensitivity in skeletal muscle. However, it remains still unclear 

how a single bout exercise enhance subsequent insulin sensitivity. The 

understanding of this mechanism would eventually contribute to further 

development of treatment for type 2 diabetes. Recently, it has been reported 

that infl ammatory M1 macrophages (MAC) are associated with development 

of insulin resistance and anti-infl ammatory M2 MAC have positive effect on 

insulin sensitivity in several insulin target organs, such as adipose tissue, 

liver and skeletal muscle. We, therefore, hypothesized that MAC, especially 

M2, are involved in the mechanisms of exercise-induced enhancement of 

insulin sensitivity in skeletal muscle. To test this hypothesis, we injected PBScontaining 

liposome (PBS) or clodronate-containing liposome (CL), a MAC 

suppressor, to C57BL6J mice. Then, mice were subjected to a single bout 

of treadmill running (20m/min, 90 min). Twenty-four hour after exercise, we 

measured ex-vivo insulin-stimulated 2-deoxy glucose (DG) uptake in skeletal 

muscle. We observed that a single bout exercise enhanced CD206 (M2 MAC 

marker)-positive MAC accumulation and insulin-stimulated 2-DG uptake in 

plantaris muscle in PBS group (Figure 1). However, CL treatment completely 

abolished MAC accumulation and enhanced insulin sensitivity in skeletal 

muscle (Figure 1). We also observed that basal AMPK phosphorylation and 

insulin-induced phosphorylation state of Akt and AS160 were not changed 

by exercise or CL treatment in plantaris muscle. From these results, we 

conclude that MAC is involved in enhancement of insulin sensitivity in 

skeletal muscle after exercise, independent of Akt and AMPK activity. 

& 1724-P 

Selective over Expression of Toll-Like Receptor 4 in Skeletal Muscle 

Causes Impaired Adaptation to High-Fat Feeding 

RYAN MCMILLAN, YARU WU, KEVIN VOELKER, JOHN KAVANAUGH, KRISTIN 

WAHLBERG, ANGELA ANDERSON, KIM HAYNIE, MORDECAI HARVEY, GABRIELLE 

FUNDARO, ELIKA SHABROKH, MADLYN FRISARD, RANDY MYNATT, MATTHEW 

HULVER, Blacksburg, VA, Baton Rouge, LA 

Our laboratory has shown that toll-like receptor 4 (TLR4) is elevated in 

skeletal muscle of obese humans and its activation results in a partitioning 

of fatty acids toward storage at the expense of oxidative pathways. To better 

understand this phenomenon, we developed a mouse model, on C57Bl/6 

background, with selective over expression of TLR4 in skeletal muscle 

(mTLR4). Mice were metabolically characterized on chow and a 45% high fat 

(HF) diet. Skeletal muscle from mTLR4 mice displayed heightened activation 

of pro-infl ammatory pathways as evidenced by increased protein levels 

of interleukin-6 and tumor necrosis factor-alpha. On a chow diet, fasting 


INTEGRATED CATEGORY 

PHYSIOLOGY—MUSCLE 

A468 

palmitate oxidation was 27% lower in red skeletal muscle from mTLR4 

mice compared to wild-type (WT); no differences were observed in white 

muscle. Following 16 weeks of HF diet, fatty acid oxidation was signifi cantly 

increased in WT mice (+17.8%, p

of genetically obese and diabetic (ob/ob) mice display a 50% size reduction 

and a dramatic lipid accumulation as compared to lean mice. To determine 

whether activation of Wnt signaling could improve lipid deposition in vivo, 

we performed a direct electrotransfection of Wnt10b cDNA or injected BIO 

in Tibialis Anterior (TA) muscles of ob/ob mice. Both up-regulated Wnt10b 

and down-regulated SREBP-1c proteins 21 days later, then inducing a 

drastic decrease in lipid deposition. Furthermore, mitochondrial Succinic 

DeHydrogenase (SDH) staining showed that Wnt signaling increased 

oxidative fi bers number by 12% ± 3% in TA of ob/ob mice, while decreasing 

glycolytic fi bers of the same amount, the size and total number of fi bers 

remaining unchanged. Immunostaining of TA slices using antibodies raised 

against the slow myosin MyhcI isoform and the fast MyhcIIA isoform 

(the most oxidative among fast isoforms) showed a 16% ± 4% increase in 

oxidative fi bers number, the number of glycolytic fi bers being decreased 

of the same amount. Our results show that activation of Wnt/β-catenin 

signaling in human and mouse skeletal muscle not only decreased lipid 

deposition, but also induced a shift towards oxydative MyHC-expressing 

fi bers, leading to improved insulin sensitivity. 

Supported by: AFM 

& 1727-P 

Mitochondrial Defi ciency of Aging Is Not Associated with Insulin 

Resistance or Higher Intramyocellular Lipid Storage 

FREDERICO G.S. TOLEDO, BRET H. GOODPASTER, PETER CHOMENTOWSKI III, 

Pittsburgh, PA 

In young adults, insulin resistance has been associated with lower 

mitochondrial oxidative capacity in skeletal muscle. This association 

suggests a defi ciency in mitochondria that may promote intramyocellular 

lipid (IMCL) deposition and insulin resistance. However, whether a defi ciency 

in mitochondria by itself is suffi cient to promote IMCL excess and insulin 

resistance in humans has not been established. Aging is associated with 

a decline in mitochondrial content and thus it is plausible that older adults 

with a decline in mitochondria should express higher IMCL accumulation and 

insulin resistance. To test this hypothesis, we studied the skeletal muscle 

mitochondrial content of younger and older adults as it relates to IMCL 

content and insulin sensitivity. Non-diabetic, middle-age (30-55 years-old) 

and older adults (>65) were divided into insulin-sensitive (IS) and resistant 

(IR) groups (Younger-IS, Younger-IR, Old-IS, and Old-IR). Euglycemic clamps 

were used to measure insulin sensitivity defi ned as glucose disposal (mg/ 

kgFFM/min). Quantitative analysis of electron micrographs of skeletal 

muscle biopsies was used to measure mitochondrial and IMCL content. In 

the IR groups, glucose disposal was nearly half of the respective IS groups 

(Younger: 5.8 vs 12.1; Old: 4.7 vs 10.7, P


Obesity 

POSTERS 

28±6% of basal blood glucose, STZ-KO 42±7% of basal blood glucose, 

p

1735-P 

Cytokine Secretion from Diabetic Skeletal Muscle: Differential 

Regulation by LPS and Fatty Acids 

THEODORE P. CIARALDI, YOUNG-SUN HONG, PAMELA TAUB, SUNDER R. MUDA- 

LIAR, ROBERT R. HENRY, La Jolla, CA, Seoul, Republic of Korea 

Chronic low-grade infl ammation has been shown to contribute to 

the development of insulin resistance, mediated in part by increased 

macrophage infi ltration into adipose tissue and skeletal muscle. The possible 

contribution of muscle cells to the production of cytokines and chemokines 

was investigated using cultured human skeletal muscle cells (hSMC) from 

healthy subjects (H, n=4) and individuals with type 2 diabetes (T2D, n=7- 

10) Myotubes were treated with lipopolysaccharide (LPS, 100 ng/mL), 

saturated (palmitate-Palm, 0.3 mM) and unsaturated (oleate,-Ol 0.3 mM) 

fatty acids or pioglitazone (Pio, 10 μM) and the media was analyzed. Under 

control conditions over 24 hrs, T2D hSMC released considerably more IL-8 

and GROα than H muscle cells, with smaller increases in IL-6 and MCP-1. In 

T2D hSMC both LPS and Palm treatment signifi cantly stimulated the release 

of IL-6 and IL-15. Palm was more potent for these effects (ex. 557 ± 71% of 

IL-6 secretion from untreated cells vs 166 ± 23% with LPS, p=0.0002), also 

signifi cantly stimulating TNFα secretion, along with a tendency to elevate 

IL-8 release. Conversely, Ol treatment signifi cantly suppressed the release 

of IL-6, IL-8 and TNFα. Pio treatment was able to reduce the secretion of 

the same cytokines, though to a lesser extent than than Ol, except in the 

case of GROα, where it was more effective (62 ± 13% of release in paired 

control cells, p


Obesity 

POSTERS 

1739-P 

Failure of Angiotensin Converting Enzyme Inhibition To Reverse 

Insulin Resistance in Female Obese Zucker Rats 

DINO PREMILOVAC, STEPHEN RATTIGAN, STEPHEN M. RICHARDS, MICHELLE A. 

KESKE, Hobart, Australia 

Impairment in insulin-mediated microvascular perfusion in muscle contri 

butes to the development of insulin resistance and type 2 diabetes. 

Angiotensin converting enzyme (ACE) inhibition has been reported to improve 

insulin sensitivity. Therefore, the aim of the current study was to determine 

whether ACE inhibition for 4 weeks improves insulin sensitivity by augmenting 

insulin-mediated microvascular perfusion in muscle. Female Obese Zucker 

(ZO) rats, at 10-12 weeks of age, were treated with quinapril (Sigma, 1mg/kg/ 

day provided in the drinking water) and compared to untreated ZO rats and 

untreated age-matched lean Zucker (LZ) rats. Following 4 weeks of treatment, 

overnight fasted anaesthetised rats were subjected to a hyperinsulinaemic 

euglycaemic clamp (20 mU/min/kg, 2hrs). Microvascular perfusion in muscle 

was assessed by metabolism of 1-methyl xanthine and muscle glucose uptake 

by 14 C-2-Deoxyglucose. After 4 weeks, untreated ZO rats had higher body 

weight (190±15 vs. 357±23 g, p

independent risk factor for diabetic heart failure. However, the underlying 

mechanisms remain poorly understood. Previous studies suggest an 

association between diabetic cardiac injury and disturbed autophagylysosome 

pathway, but the specifi c changes and their signifi cance are not 

fully characterized. Here, we show that the expression and distribution of 

Cathepsin D, an aspartyl protease normally restricted within lysosome, 

were dramatically altered in cultured cardiomyocytes exposed to high 

glucose. Specifi cally, Western blotting and RT-PCR showed that high glucose 

(17 or 30 nmol/l) markedly increased the protein and mRNA expression levels 

of Cathepsin D in cardiomyocytes. Moreover, immunostaining analysis 

indicated that Cathepsin D lost its punctate distribution and was diffused in 

cardiomyocytes after high glucose treatment. To investigate the functional 

signifi cance of altered Cathepsin D in hyperglycemic toxicity, we treated 

cardiomyocytes with a low dose of the lysosome inhibitor Bafi lomycin A1 

(BFA, 0.2nmol/l). Remarkably, BFA inhibited Cathepsin D maturation and 

attenuated high glucose-induced myocyte injury, as shown by reduced 

cleavage of Poly (ADP-ribose) polymerase (PARP). Together, these fi ndings 

suggest that increased expression and altered distribution of Cathepsin 

D may contribute to hyperglycemic cardiotoxicity. Consistently, we found 

that Cathepsin D was also increased in the hearts of type 1 diabetic mice 

(Streptozotocin induced or OVE26 genetic model). Thus, future studies are 

warranted to determine if Cathepsin D plays a similar role in diabetic cardiac 

damage in vivo. 



INTEGRATED CATEGORY 

PHYSIOLOGY—MUSCLE 


1744-P 

Increased Inducible Nitric Oxide Synthase Activity Is Responsible 

for Reduced Endothelin-1 Vasoconstrictor Responsiveness in Insulin 

Resistance 

CAROL T. BUSSEY, MICHELLE A. KESKE, STEPHEN RATTIGAN, STEPHEN M. 

RICHARDS, Hobart, Australia 

Increased levels of the potent vasoconstrictor endothelin-1 (ET-1) have 

been implicated in the development of insulin resistance, type 2 diabetes 

and hypertension, and particularly the endothelial dysfunction seen in 

these states. Endogenous ET-1 activity is consistently increased in these 

pathologies, but opinion is divided as to whether ET-1 vasoconstrictor 

responsiveness is enhanced or impaired. Our study examined the sensitivity 

of skeletal muscle vasculature to ET-1 in rats made insulin resistant by high 

fat feeding, using the isolated, pump-perfused hindlimb preparation. Male 

Sprague-Dawley rats were fed a high-fat diet for 4 weeks, resulting in an 

81% increase in epididymal fat pad weight, accompanied by increased 

fasting plasma insulin, but not glucose. Vasoconstriction by ET-1 (1 or 3nM) 

was reduced by the high-fat diet. Basal perfusion pressure was unaffected. 

Treatment with the nitric oxide (NO) synthase (NOS) inhibitor N G -nitro-Larginine 

methyl ester (L-NAME) signifi cantly increased ET-1 vasoconstriction 

in both normal and high-fat fed animals, reaching the same level, indicating 

increased NO bioavailability in insulin resistance. In the presence of 1400W, 

a specifi c inhibitor of the inducible NOS (iNOS) isoform, ET-1 reactivity in highfat 

fed rats was restored to that from rats on normal chow. These fi ndings 

support the notion that high fat feeding increases muscle iNOS activity, 

counteracting ET-1 vasoconstriction. Alterations in the balance between 

constriction and NO-mediated dilation may account for differences observed 

in ET-1 responsiveness in metabolic syndrome pathologies. Furthermore, 

the raised NO production in high fat fed rats may reduce responsiveness to 

further NO-mediated dilation, and may explain the impairment of endothelial 

function seen in insulin resistant states. 

A473 

1745-P 

Losartan Recruits Muscle Microvasculature and Prevents Lipid- 

Induced Metabolic Insulin Resistance 

NASUI WANG, WEIDONG CHAI, EUGENE J. BARRETT, ZHENQI LIU, Charlottesville, 

VA 

Patients with diabetes have increased plasma concentrations of free fatty 

acids (FFAs) which cause insulin resistance in multiple insulin responsive 

tissues, including the microvasculature. Microvascular insulin resistance 

contributes signifi cantly to metabolic insulin resistance seen in diabetes. 

Blockade of angiotensin II type 1 receptor with losartan recruits muscle 

microvasculature and may improve insulin sensitivity. Whether losartan 

recruits muscle microvasculature and rescues insulin’s metabolic action in 

the presence of high FFA concentrations is not known. 

After an overnight fast, male Sprague-Dawley rats received a systemic 

infusion of either saline or intralipid + heparin (3.3% and 30 U/ml) for 3 

hrs after an overnight fast, with a 3 mU/min/kg euglycemic insulin clamp 

superimposed for the last 2 hrs. A third group received the same infusion 

of intralipid + heparin and insulin, with a losartan injection (0.3 mg/kg, i.v. 

bolus) 5 min before starting the insulin infusion. Muscle microvascular blood 

volume (MBV) and microvascular fl ow velocity (MFV) were measured using 

contrast-enhanced ultrasound before and at 30, 60 and 120 min of insulin 

infusion. Muscle microvascular blood fl ow (MBF) was calculated as the 

product of MBV and MFV. Steady state whole body glucose disposal rates 

were calculated. 

Insulin infusion doubled muscle MBV at 30 min and this effect lasted 

for the entire 120 min (p


Obesity 

POSTERS 

to IMTG accumulation. High FFA may compromise mitochondrial function, 

termed mitochondrial lipotoxicity, which could contribute to insulin 

resistance. Thus, we determined mitochondrial function during short-term 

elevation of circulatory FFA. 

We included 12 young, healthy sedentary humans (8 males, 4 females; 

age: 29±2 yrs; BMI: 23.4±0.6 kg/m 2 ) who underwent 4 hours of Intralipid 

(20%) infusion followed by an euglycemic-hyperinsulinemic clamp to assess 

insulin sensitivity. Detailed mitochondrial function, including ADP-coupled 

substrate oxidation on glutamate and succinate (state 3), FCCP-induced 

maximal oxidative capacity (state u) as well as enzyme kinetics (K m and V max ) 

of NADH and FADH 2 dehydrogenase, was examined in permeabilized muscle 

fi bers using high-resolution respirometry. Muscle biopsies from the m.vastus 

lateralis were taken before (baseline) and at 4 hours of intralipid infusion. 

Elevation of plasma FFA (from 0.48±0.05 to 2.12±0.19 mmol/l, p

MCK-PPARγ were compared to related wildtype mice (CON). Weight gain 

was 32% less in female MCK-PPARγ versus CON (p


Obesity 

POSTERS 

actions in vivo, we generated transgenic mice which specifi cally express the 

gene coding for adiponectin receptor 1 (AdR1) in mouse macrophages using 

the scavenger receptor A-I gene (SR-AI) enhancer/promoter. 

Analyzing this transgenic mouse model, we have determined that AdR1 

expression in macrophages was associated with a signifi cantly decreased 

cholesterol (45%, p

& 1759-P 

Dissecting the Intestinal Cholescystokinin Signaling Pathway(s) 

That Regulate Glucose Production 

BRITTANY A. RASMUSSEN, DANNA M. BREEN, GRACE W.C. CHEUNG, ANDREA 

KOKOROVIC, TONY K.T. LAM, Toronto, ON, Canada 

Diabetes and obesity is characterized by a disruption in glucose 

homeostasis due partly to an elevation of hepatic glucose production. To 

date, the mechanisms underlying the regulation of glucose production in 

healthy and obese/diabetic settings remain to be elucidated. Duodenal lipid 

metabolism is recently demonstrated to trigger a gut-brain-liver neuronal 

axis to lower glucose production in rats in vivo. The gut derived hormone 

cholecystokinin (CCK) mediates this gut lipid neuronal effect through the 

activation of CCK-A receptors. However, duodenal lipid-CCK activation fails 

to lower glucose production in response to high-fat feeding, suggesting 

duodenal CCK resistance. To begin locating the molecular signaling defect(s) 

of duodenal CCK-resistance, we fi rst discovered that high-fat feeding did 

not alter duodenal CCK-A receptor expression, suggesting that duodenal 

CCK resistance occurs at the signaling level of the CCK-A receptors. We 

next investigated whether the activation of PKA, a downstream signaling 

molecule of the CCK-A receptor, is suffi cient and necessary for duodenal CCK 

to lower glucose production through a neuronal network in normal rats in 

vivo. First, intraduodenal infusion of the PKA activator, Sp-CAMPS, lowered 

glucose production during the pancreatic (basal insulin) euglycemic clamps. 

Co-infusion of Sp-CAMPS with the PKA inhibitor H89 intraduodenally 

abolished the glucose production suppression effect. Second, intraduodenal 

administration of CCK-8 (the biologically active form of CCK) lowered glucose 

production. Importantly, co-administration of CCK-8 with H89 abolished the 

ability of duodenal CCK-8 to lower glucose production. Third, the ability of 

Sp-CAMPS to lower glucose production was abolished upon intraduodenal 

co-administration of the anesthetic tetracaine with Sp-CAMPS. In Summary, 

these data together illustrate that duodenal PKA activation is suffi cient and 

necessary to lower glucose production via a neuronal network in normal 

rats. This set of preliminary data raises the possibility that high fat feeding 

induces duodenal CCK resistance at the signaling level of duodenal PKA. 

Supported by: CIHR 

& 1760-P 

Gastrointestinal-Mediated Glucose Disposal in Vagotomized Subjects 

ASTRID PLAMBOECK, TINA VILSBØLL, CAROLYN DEACON, ANDRE WETTERGREN, 

SØREN MEISNER, CLAUS HOVENDAL, LARS BO SVENDSEN, FILIP KNOP, JENS 

HOLST, Hellerup, Denmark, Copenhagen, Denmark, Odense, Denmark 

After secretion, glucagon-like peptide-1 (GLP-1) is rapidly degraded by 

the enzyme dipeptidyl peptidase-4 (DPP-4), resulting in less than 15% of 

the intact biologically active peptide reaching the systemic circulation. This 

has led to the hypothesis that GLP-1 acts locally before being degraded. We 

aimed to evaluate the role of the vagus nerves for the effect of GLP-1 on 

gastrointestinal-mediated glucose disposal (GIGD). 

Eight truncally vagotomized (due to duodenal ulcer) subjects (age: 70±2 

years (mean±SEM); BMI: 24±1 kg/m 2 ; fasting plasma glucose (FPG): 5.8±0.2 

mM), 7 subjects previously treated for esophageal cancer with resection 

of the cardia including truncal vagotomy (age: 64±2 years; BMI: 23±1 kg/ 

m 2 ; FPG: 5.4±0.1 mM) and 5 healthy control subjects (age: 68±2 years; BMI: 

25±1 kg/m 2 ; FPG: 5.4±0.2 mM) were examined on three separate occasions: 

two 4h 50-g oral glucose tolerance tests (OGTTs) ± DPP-4 inhibition and an 

isoglycemic intravenous (iv) glucose infusion (IIGI). 

Isoglycemia during IIGIs was obtained using 24±2 g and 28±4 g of glucose 

in patients with truncal vagotomy associated with surgery for duodenal 

ulcer and cardia resection, respectively (P=NS) and 17±2 g in healthy control 

subjects (P


Obesity 

POSTERS 

Guided Audio Tour: Incretin Hormone Biology (Posters 1763-P to 1772-P), 

see page 13. 

1763-P 

Chronic Administration of Ezetimibe Increases Active Glucagon-Like 

Peptide-1 and Prevents the Development of Type 2 Diabetes in Rats 

SOO JIN YANG, JUNG MOOK CHOI, LISA KIM, WON JUN KIM, SE EUN PARK, 

EUN JUNG RHEE, CHEOL-YOUNG PARK, WON YOUNG LEE, KI WON OH, SUNG 

WOO PARK, SUN WOO KIM, Seoul, Republic of Korea 

Ezetimibe is a cholesterol-lowering agent targeting Niemann-Pick C1like 

1, an intestinal cholesterol transporter. Chronic administration of 

ezetimibe may ameliorate several metabolic disorders including hepatic 

steatosis and insulin resistance. In this study, we investigated whether 

chronic ezetimibe treatment prevents the development of type 2 diabetes 

and alters levels of glucagon-like peptide-1 (GLP-1), an incretin hormone 

involved in glucose homeostasis. Male LETO and OLETF rats at 12 weeks of 

age were treated with vehicle or ezetimibe (10 mg·kg-1 · day-1 WITHDRAWN 

) for 20 weeks 

via stomach gavage. OLETF rats were diabetic with hyperglycemia and 

signifi cant decreases in pancreatic size and beta cell mass compared with 

lean controls. There was no difference in weight change and food intake 

between groups during the treatment period. However, chronic treatment of 

the OLETF rats with ezetimibe prevented the development of diabetes with 

reduced fasting serum glucose (7.2 ± 0.1 vs. 10.4 ± 0.2 mmol/l, P < 0.001) 

and improved glucose control during oral glucose tolerance test (28504 ± 

1486 vs. 37512 ± 2052 area under the curve, P = 0.002) compared with OLETF 

controls. Moreover, ezetimibe treatment rescued the reduced pancreatic 

size and beta cell mass in OLETF rats. Consistent with the previous fi nding 

that diabetic patients showed high dipeptidyl peptidase-4 (DPP-4) activity 

and low levels of active GLP-1, DPP-4 activity was higher and active GLP- 

1 tended to be lower in diabetic OLETF rats compared with lean controls. 

Interestingly, ezetimibe signifi cantly decreased serum DPP-4 activity (OLETF 

ezetimibe 309.5 ± 7.3, OLETF control 478.2 ± 29.7 % LETO control; P < 0.001) 

and increased active GLP-1 (OLETF ezetimibe 14.8 ± 4.2, OLETF control 5.3 

± 0.3 pmol/l; P = 0.034) in OLETF rats. These fi ndings demonstrated that 

chronic administration of ezetimibe prevented the development of type 2 

diabetes and increased active GLP-1 levels, suggesting possible involvement 

of GLP-1 in the preventive effect of ezetimibe against type 2 diabetes. 

& 1764-P 

Prohormone Convertase 2 Positive Enteroendocrine Cells Are More 

Abundant in Patients with Type 2 Diabetes—A Potential Source of 

Gut-Derived Glucagon 

FILIP K. KNOP, KRISTINE J. HARE, JENS PEDERSEN, JAKOB W. HENDEL, STEEN S. 

POULSEN, JENS J. HOLST, TINA VILSBØLL, Hellerup, Denmark, Copenhagen, Denmark, 

Herlev, Denmark 

In α cells, the precursor proglucagon (from the glucagon gene) is 

processed by prohormone convertase (PC)2 to glucagon, whereas enteroendocrine 

L cells utilize PC1 in the processing of proglucagon to the 

glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). Hyperglucagonemia 

following oral glucose in type 2 diabetes mellitus (T2DM) is thought 

to arise as a consequence of dysfunctional α cells combined with β cell 

insuffi ciency. However, in contrast to oral glucose, iv glucose does not elicit 

hypersecretion of glucagon in T2DM. Therefore, we hypothesized that T2DM 

patients possess the potential to release glucagon directly from the gut. 

Ten male patients with T2DM (age: 51(41-62) years); BMI: 32(28-39) kg/m 2 ; 

HbA1c: 7.1(5.4-8.7)%) and 10 male healthy control subjects (age: 58(48-67) 

years; BMI: 31(26-36) kg/m 2 ; HbA1c: 5.5(5.2-6.0)%) underwent a 4h meal test 

and a jejunoscopy (including jejunal biopsies) on two separate days. 

Patients with T2DM exhibited exaggerated postprandial plasma glucose 

excursions (379±76 (mean±SEM) vs. 77±33 mM×4h, P=0.001). Postprandial 

insulin (30±6 vs. 27±5 nM×4h, P=0.7) and C-peptide responses (175±25 

vs. 188±24 nM×4h, P=0.7) were similar in the two groups, but patients 

with T2DM exhibited higher postprandial glucagon responses (3.0±0.5 vs. 

1.9±0.2 nM×4h, P=0.02). No differences in glucose-dependent insulinotropic 

polypeptide (GIP), GLP-1, GLP-2 or peptide YY responses were observed. 

Similar numbers of endocrine cells (all stained for PC1) from jejunal biopsies 

were observed in the two groups; including GIP, GLP-1, and GLP-2 positive 

cells. Signifi cantly more PC2 positive cells were found among T2DM patients 

(70±8 vs. 44±4 cells/mm 2 , P=0.01). Similar levels of PC1 and PC2 gene 

expression were observed in the two groups. 

Our results show that a high number of small intestinal endocrine cells in 

T2DM patients are equipped with PC2, which potentially - through processing of 

proglucagon to glucagon - contribute to the hyperglucagonemia of these patients; 

shifting the ‘pancreacentric’ view on type 2 diabetic hyperglucagonaemia 

towards a role for the gut in this pathophysiological trait. 


INTEGRATED PHYSIOLOGY—OTHER CATEGORY HORMONES 

A478 

& 1765-P 

Glucose-Dependent Insulinotropic Polypeptide—A Bifunctional Blood 

Glucose Stabilizer 

MIKKEL CHRISTENSEN, LOUISE VEDTOFTE, JENS J. HOLST, TINA VILSBØLL, FILIP 

K. KNOP, Hellerup, Denmark, Copenhagen, Denmark 

Longstanding knowledge positions glucose-dependent insulinotropic 

polypeptide (GIP) as an incretin hormone in healthy humans, but controversy 

exists regarding the effects of GIP on glucagon secretion. We hypothesized 

that the glucagonotropic effect of GIP, like its insulinotropic effect, is 

glucose-dependent. Therefore, we aimed to evaluate the effect of GIP on 

plasma glucagon and insulin responses at three different glycemic levels. 

Ten healthy male subjects (age: 23±1 (mean±SEM) years; BMI: 22±1 

kg/m 2 ; HbA 1 c: 5.5±0.1%) without family history of diabetes were studied on 

six separate days. Physiological doses of GIP or saline were administered 

intravenously (randomized and double-blinded) during 90 min of either 

insulin-induced hypoglycemia, euglycemia or hyperglycemia (randomized). 

During hypoglycemia plasma glucose (PG) was gradually lowered from 

mean fasting level of 5.0±0.1 mM (mean±SEM) to a plateau level of 2.8±0.1 

mM. GIP infusion resulted in greater glucagon responses during the fi rst 30 

minutes compared to saline (area under curve: 76±17 vs. 28±16 pM×30 min, 

P

Nadir glucose 

(mmol/L) 

Time to reach nadir 

glucose (min) 


Hypoglycemic-GB (H) Asymptomatic-GB (A) Statistical tests (p values) 

Saline Ex-9 Saline Ex-9 H vs. A Ex-9 vs. Saline Interaction 

2.3±0.5 4.1±0.4 3.9±0.3 4.2±0.2 0.19 0.00 0.01 

79±3 146±25 120±30 180±0 0.15 0.01 0.84 

AUCglucose (mmol/L.min) 632±151 994±51 970±42 1056±73 0.16 0.03 0.12 

AUCinsulin (µmol/L.min) 

59.4±14.7 30.6±3.6 40.1±16.2 28.6±11.9 0.40 0.11 0.44 

These fi ndings indicate that enhanced GLP-1 action contributes to 

postprandial hypoglycemia after GB and GLP-1 receptor antagonists are an 

attractive potential treatment for this condition. 

Supported by: NIH-DK083554 

& 1767-P 

Glucose-Dependent Insulinotropic Polypeptide Suppresses the 

Development of Atherosclerosis in Apolipoprotein E-Null Mice Via 

Its Own Receptors 

MASAHARU NAGASHIMA, MICHISHIGE TERASAKI, KYOKO NOHTOMI, MASAKO 

TOMOYASU, SHURI KANEYAMA, TAKUYA WATANABE, AKIRA MIYAZAKI, 

TSUTOMU HIRANO, Shinagawa, Japan, Hachiouji, Japan 

Several reports have suggested glucagon-like peptide-1 has favorable 

effects on the suppression of atherosclerosis. Less is understood, however, 

on how atherosclerosis is affected by glucose-dependent insulinotropic 

polypeptide(GIP). We report experimental evidence demonstrating that GIP 

has direct anti-atherosclerotic properties. 

Starting from the age of 17 weeks, apolipoprotein (apo) E-null mice were 

placed on an atherogenic diet and administered daily subcutaneous infusions 

of vehicle (saline), GIP(1-42) (25 nmol/kg/day), GIP(3-42) (25nmol/kg/day), 

(Pro3)GIP (25nmol/kg/day), or GIP(1-42) plus (Pro3)GIP for 4 weeks. The 

infusions did not signifi cantly affect body weight, food intake, plasma lipids, 

or glucose. GIP(1-42) suppressed the atherosclerotic lesions in the whole 

aorta by 53%, suppressed the plaque volume in the aortic valve by 38%, 

and suppressed macrophage infi ltration by 48%, compared with the vehicle 

controls. In contrast, GIP(3-42) had no effects on these atherosclerotic 

changes, and the co-infusion of (Pro3)GIP abolished the anti-atherosclerotic 

effect of GIP(1-42). The infusion of (Pro3)GIP alone had no effect on the 

development of atherosclerosis. 

An oxidized LDL-induced foam cell formation was detected in the peritoneal 

macrophages obtained from apo E-null mice with various infusions. GIP(1- 

42) infusion suppressed the foam cell formation by 30%, whereas GIP(3-42) 

infusion had no suppressive effect whatsoever. Meanwhile, the co-infusion 

with (Pro3)GIP nullifi ed the suppressive effect of GIP(1-42). 

GIP receptor mRNA was detected in human and murine macrophages, 

vascular smooth muscle cells, and endothelial cells by RT-PCR. GIP receptors 

were also histochemically stained in the vascular endothelium, media, and 

plaque of aorta in the apo E-null mice. 

GIP (1nM) in vitro suppressed the proliferation of smooth muscle cells 

determined by BrdU staining, the gene expressions of ICAM-1 and MCP-1 in 

endothelial cells, and macrophage foam cell formation. 

These results suggest that GIP has direct anti-atherosclerotic effects on 

the cells mainly responsible for atherogenesis via its own receptors. 

& 1768-P 

Antidiabetic Effect of Inhibitors of Apical Sodium-Dependent Bile 

Acid Transport (ASBT) 

XIAOZHOU YAO, JUDI A. MCNULTY, DONALD I. ANDERSON, YAPING LIU, 

CHRISTOPHER C. NYSTROM, DALLAS K. CROOM, SEAN A. ROSS, DEEPAK K. 

RAJPAL, KIMBERLY D. HAMLET, JON L. COLLINS, CHARI D. SMITH, ANDREW A. 

YOUNG, LIHONG CHEN, Research Triangle Park, NC 

Bile acids are increasingly recognized as metabolic modulators. We 

evaluated the effects on glucose homeostasis of a potent ASBT inhibitor 

(264W94), which blocks intestinal absorption of bile acids and increases 

fecal bile acid concentration in Zucker Diabetic Fatty (ZDF) rats. Drug was 

orally administered twice daily for two weeks in doses of 0.001, 0,01, 0,1, 1 

and 10 mg/kg (n=8/dose group). Non-fasting blood glucose, HbA1c, insulin, 

plasma total glucagon-like peptide 1 (tGLP-1) and bile acid concentrations 

and fecal bile acids were measured weekly. An oral glucose tolerance 

test (OGTT) was performed at the end of the study. Oral administration of 

264W94 dose-dependently decreased plasma bile acids, and increased fecal 

bile acid and non-fasting plasma tGLP-1 throughout the course of the study. 

Treatment with 264W94 signifi cantly decreased HbA1c and glucose, and 



A479 

prevented the drop of insulin levels in a dose-dependent manner. Effective 

doses increased GLP-1 up to 2-fold and insulin up to 3-fold during the OGTT. 

Administration of selective agonists at bile acid responsive receptors, 

the farnesoid-X receptor and TGR5, did not replicate the effects of ASBT 

inhibition, suggesting an alternate mechanism. In summary, inhibition of 

ASBT increases bile acids in the distal intestine, promotes GLP-1and insulin 

release and glucose-lowering, and may offer a new therapeutic strategy for 

type 2 diabetes mellitus. 

& 1769-P 

Novel Biological Action of the Dipeptidylpeptidase 4 Inhibitor, 

Sitagliptin, as a GLP-1 Secretagogue 

GANESH V. SANGLE, LINA M. LAUFFER, ANTHONY GRIECO, PATRICIA L. 

BRUBAKER, Toronto, ON, Canada 

Glucagon-like peptide-1 (GLP-1) is released from the intestinal L cell in 

response to nutrients and other secretagogues. GLP-1 has important antidiabetic 

effects including enhancement of glucose-dependent insulin 

secretion. Sitagliptin prevents GLP-1 degradation by inhibiting dipeptidylpeptidase 

4 (DPP4), and is used to lower HbA1c levels in patients with 

type 2 diabetes (T2D). Sitagliptin treatment for 4 wk increased bioactive 

GLP-1 levels by 80% and improved glycemic tolerance by 60% in neonatal- 

STZ rats, a T2D model. This effect was prevented by the GLP-1 antagonist, 

exendin-4(9-39NH2), confi rming GLP-1-dependence of the actions of 

sitagliptin. However, it is unclear if sitagliptin increases GLP-1 levels only 

via DPP4 inhibition or if it has actions on GLP-1 independent of DPP4. The 

effects of sitagliptin (0.1-2μM) directly on the L cell were thus tested using 

the murine (GLUTag) and human (NCI-H716) L cell lines, as well as primary 

fetal rat intestinal L cells (FRIC). Sitagliptin increased total GLP-1 secretion in 

the GLUTag and NCI-H716 cells, by up to 132 and 83%, resp. (P


Obesity 

POSTERS 

Pre-administration of excess non-radioactive exendin(9-39) signifi cantly 

blocked the radioactivity of pancreas after [ 125 I]IB-Ex(9-39) injection. SPECT 

was performed 30 min after [ 123 I]IB-Ex(9-39) administration to mice, which 

revealed the remarkable accumulation of radioactivity in pancreas. These 

results indicated that Ex(9-39) could serve as a useful probe for non-invasive 

SPECT imaging of pancreatic β-cells. Grant Acknowledgement: The Program 

for Promotion of Fundamental Studies in Health Sciences of the National 

Institute of Biomedical Innovation (NIBIO). 

& 1771-P 

Basal Plasma Glucose, Insulin and Glucose-Dependent Insulinotropic 

Polypeptide Constitute Important Determinants of Fasting 

Hyper glucagonemia in Type 2 Diabetes 

JONATAN I. BAGGER, FILIP K. KNOP, MAI-BRITT T. NIELSEN, JENS J. HOLST, 

TINA VILSBØLL, Hellerup, Denmark, Hvidovre, Denmark, Copenhagen, Denmark 

Fasting hyperglucagonemia in patients with type 2 diabetes mellitus 

(T2DM) contribute to the exaggerated fasting plasma glucose (FPG) levels of 

these patients. However, the mechanisms underlying elevated basal plasma 

glucagon levels are poorly understood. We aimed to clarify the relationship 

between basal glucagon levels and several metabolic parameters including 

the gut incretin hormones glucose-dependent insulinotropic polypeptide 

(GIP) and glucagon-like peptide-1 (GLP-1). 

Blood from patients with T2DM (N=104, 27% women; age: 57±1 years 

(mean±SEM); body mass index (BMI): 30±1 kg/m 2 ; FPG: 10.8±0.3 mmol/L; 

HbA 1 c: 8.0±0.2%; diabetes duration: 53±6 months) and healthy control 

subjects (N=71, 27% women; age: 56±1 years; BMI: 28±1 kg/m 2 ; FPG: 

5.6±0.1 mmol/L (P

insulin, glucose, adiponectin, and plasma apelin (p = 0.011). In conclusion, 

apelin expression in adipose tissue is associated with insulin sensitivity as 

assessed by SSPG. This association is independent of plasma apelin levels, 

and suggests that apelin’s impact on systemic insulin sensitivity may be 

infl uenced by its effects in adipose tissue. 

Supported by: NIH: 1K08DK080463 (to PY), 5R01DK071333 (to PST), 5R01DK071309 

(to GMR) 

1775-P 

B-Type Natriuretic Peptide Affects the Response to Intravenous 

Glucose in a Placebo-Controlled Cross-Over Study in Healthy Volunteers 

MICHAEL RESL, BIRGIT HEINISCH, GREISA VILA, MICHAELA RIEDL, EVELYNE 

WOHLSCHLAEGER-KRENN, GIOVANNI PACINI, MARTIN CLODI, ANTON LUGER, 

Vienna, Austria 

B-type natriuretic peptide (BNP) is a hormone secreted from the heart 

in response to volume load and serves clinically to help reduce the cardiac 

work load. BNP is as well a reliable biomarker in the diagnosis of cardiac 

dysfunction and heart failure. As patients with heart failure present an 

increased risk for developing diabetes, we aimed to investigate the role 

of BNP on parameters of glucose metabolism in a placebo-controlled 

crossover study performed in 10 healthy volunteers (25±1 years; BMI 23±1 

kg/m 2 ; fasting glucose 83± mg/dl). Participants received intravenously either 

placebo or 3 pmol/kg/min BNP-32 for 4h. One hour after beginning the BNP/ 

placebo infusion, a 3h intravenous glucose tolerance test (0.33 g/kgglucose 

+ 0.03 U/kg insulin at 20 min) was started. Plasma glucose, insulin and 

C-peptide were frequently measured for minimal model analysis. 

BNP increased the glucose distribution volume (13±1 %BW vs. 11±1, 

P


Obesity 

POSTERS 

metabolic controls of appetite/body weight. We found that oxytocin release 

displayed distinct circadian rhythmicity in normal mice, with hypothalamic 

and circulating levels of oxytocin gradually increased during the daytime and 

drastically declined during the nighttime. The circadian release of oxytocin 

was found to underlie the temporal patterns of appetite in a negative 

manner in normal chow-fed mice. In contrast, chronic high-fat diet (HFD) 

feeding impaired oxytocin release predominantly during the daytime, leading 

to vanished circadian rhythms of oxytocin release. This change underlied the 

prominent promotion of appetite by HFD during the daytime rather than during 

the nighttime. To explore molecular mediators, we examined the relationship 

between oxytocin and synaptotagmin 4 (Syt4), suggested by our recent work 

showing that Syt4 is an exocytotic inhibitor of oxytocin release (Zhang et al, 

Neuron, in press). Data revealed that Syt4 localization in oxytocin vesicles 

was >10-fold less active during the daytime than that during the nighttime. 

Consistently, HFD feeding elevated Syt4 localization in oxytocin vesicles 

predominantly during the daytime, leading to the circadian arrhythmicity of 

oxytocin release. Physiological studies using Syt4 knockout mice revealed 

that Syt4 inhibition prevented HFD-induced circadian arrhythmicity in oxytocin 

release and appetite, which accounted for the anti-obesity phenotype of 

Syt4 knockout mice. In conclusion, Syt4-directed oxytocin release represents 

a hypothalamic neuropeptide program that mediates circadian regulation/ 

dysregulation of energy and body weight balance. 

Supported by: NIH ADA-Funded Research 

1780-P 

Circulating Plasma Pigment Epithelium-Derived Factor (PEDF) Levels 

Are Associated with Insulin Resistance in Women with Estrogen 

Defi ciency 

MICHELE M.A. YUEN, WING SUN CHOW, CHEN CHENG, TING CHUNG PUN, 

LAW RENCE S.C. LAW, ANNETTE W.K. TSO, AIMIN XU, KAREN S.L. LAM, Hong 

Kong, China 

Pigment epithelium-derived factor (PEDF) is implicated in murine insulin 

resistance and metabolic dysfunctions. Women have lower serum PEDF levels 

and 17β-estradiol inhibits the transcription of PEDF in human ovarian epithelial 

cells. To investigate whether estradiol regulates PEDF in vivo, we studied the 

changes in PEDF levels in 21 pre-menopausal women (age = 49.6 ± 3.5 years) 

following bilateral oophorectomy for benign gynaecological conditions. 

Body mass index (BMI), waist circumference (WC), percentage body fat, 

and plasma PEDF (ng/ml) and estradiol (pmol/l) levels were measured before 

and after oophorectomy. The changes (Δ) in PEDF, estradiol, homeostatic 

model assessment of insulin resistance (HOMA-IR) and quantitative insulinsensitivity 

check index (QUICKI) were analyzed with repeated measure 

ANOVA. The mean duration between pre- and post-operative assessment 

was 4.0±0.7 months. BMI was marginally reduced in the post-operative 

period (25.2 ± 4.0 kg/m 2 pre-op vs. 24.8 ± 4.3 kg/m 2 post-op; p = 0.048). There 

were no signifi cant changes in WC or percentage body fat The marked postoperative 

reduction in plasma estradiol levels (225 [109-418] pmol/l pre-op 

vs. 32 [19-50] pmol/l post-op; p < 0.001) was associated with increases in 

plasma PEDF levels (7.7 ± 1.6 ng/ml pre-op vs. 8.8 ± 1.5 ng/ml post-op; p = 

0.001). An inverse relationship was found between Δestradiol and ΔPEDF (r = 

-0.497; p = 0.022). Insulin resistance increased post-operatively, as indicated 

by HOMA-IR (1.06 [0.73-1.93] pre-op vs. 1.63 [1.06-2.26] post-op; p = 0.002) 

and QUICKI (0.38 ± 0.04 pre-op vs. 0.36 ± 0.03 post-op; p = 0.006). ΔHOMA- 

IR and ΔQUICKI became insignifi cant (p = 0.101 and 0.141 respectively) after 

adjusting for ΔPEDF. In conclusion, the reduction in plasma estradiol levels 

following oophorectomy was signifi cantly associated with increases in 

plasma PEDF levels and insulin resistance. Our data suggest that estradiol is 

a physiological regulator of PEDF expression in humans and that an increase 

in circulating PEDF contributes to the increase in insulin resistance in these 

women with estrogen defi ciency following bilateral oophorectomy. 

1781-P 

Decreased Aortic Angiotensin Converting Enzyme 2 (ACE 2) and 

Neprilysin (NEP) Protein Expression in db/db Diabetic Mice 

HARSHITA CHODAVARAPU, NARGES KABLAN, RENDONG QUAN, KHALID 

ELASED, Dayton, OH 

Cardiovascular disease is a long term complication of diabetes, which 

remains a leading cause of morbidity and mortality. There is evidence of 

activation of renin angiotensin system (RAS) in diabetic animals and humans. 

Emerging data shows that the vasoconstrictor actions of Angiotensin II (Ang 

II) may be opposed by formation of the vasodilator, Ang (1-7). ACE2 recently 

was identifi ed as a homologue of ACE that preferentially forms Ang-(1-7) from 

Ang II. Neutral endopeptidase (NEP), a cell surface metallopeptidase also 



A482 

generates Ang (1-7) by degrading Ang I. Ang (1-7) mediates its vasodilatation 

action through a G protein-coupled receptor (Mas). In our previous studies 

we demonstrated an age-dependent increase of blood pressure in db/db 

type 2 diabetic mice. The goal of this study was to evaluate whether there is 

an age-dependent changes in aortic ACE, ACE2 and NEP protein expression 

in db/db diabetic mice. Plasma ACE and ACE2 activities were measured. 

Western blot analysis demonstrated a signifi cant decrease in aortal ACE2 

and NEP protein expression in 12 wk hypertensive db/db mice compared to 

controls (p

control and one preceded by a sham feeding (chewing and expectorating 

peanut butter sandwich, 255kcal). Areas under the curves (AUC) for glucose 

and insulin response to meal ingestion over the premeal values were 

compared using two-way repeated ANOVA. 

H subjects had lower nadir glucose, lower AUC glucose , and higher AUC insulin 

values compared to A group during control studies. Sham feeding had no effect 

on the glucose and insulin responses in the H subjects. However, in A subjects 

sham feeding caused a signifi cant decrease in AUC glucose (p


Obesity 

POSTERS 

1787-P 

Implication of Corticotropin-Releasing-Hormone on INS-1 and Pancreatic 

Islets In Vitro 

BARBARA LUDWIG, JANINE SCHMID, CHRISTIAN ZIEGLER, MONIKA EHRHART- 

BORNSTEIN, STEFAN R. BORNSTEIN, Dresden, Germany 

Hormonal factors, including corticotropin-releasing-hormone (CRH) and 

glucocorticoids (GC) regulate the activity of the hypothalamic-pituitaryadrenal 

(HPA) axis in response to stress. This axis is kept in balance by 

the negative feedback effects of GC on the CRH synthesis and secretion 

in the hypothalamus. Recently CRH has been identifi ed to promote β-cell 

proliferation and potentiates insulin secretion in a glucose dependent 

manner. On the other hand, GCs are referred to as diabetogenic hormones 

due to the induction of gluconeogenesis, implication on development of 

insulin resistance, effects on adipocytes and the functionally insulinantagonizing 

effects. Therefore, an imbalance of the CRH/GC system may 

lead to metabolic dysfunction and diabetes. 

GC access to intracellular receptors is regulated by two isoforms 

of 11β-hydroxysteroid dehydrogenase (11β-HSD) which catalyse the 

interconversion of physiologically active GC to its inactive metabolite 

(11β-HSD-2) and vice versa (11β-HSD-1). 

In the present study we analyzed the mechanism of CRH mediated 

β-cell regulation and asked if pancreatic islets respond directly to CRH by 

interference with 11β-HSD activity and therefore GC activity. 

In in vitro studies we could demonstrate that CRH and its receptor are 

expressed on mRNA and protein levels in INS-1 cells, rat and human islets. 

We also found expression of both 11β-HSD isoforms in rat and human 

islets. CRH exposure of islets signifi cantly decreased mRNA levels of 

both 11β-HSD-isoforms as measured by quantitative RT-PCR. The specifi c 

iso-enzyme activity was analyzed by measuring the production of active 

GC. Following CRH treatment, active GC levels were signifi cantly reduced 

indicating an overbalance in favour of 11β-HSD-2 activity. Stimulation with 

CRH resulted in signifi cantly increased insulin secretion. Moreover, CRHreceptor 

activation caused a signifi cant increase of cell proliferation and 

reduced cell apoptosis. 

We suggest that CRH may not only be of signifi cance within the 

endocrine stress system for triggering and sustaining obesity and metabolic 

dysfunction, but may also play a direct role for glucose homoeostasis and 

the regulation of β-cell mass. 

1788-P 

Improved Glycemic Control Enhances the Incretin Effect in Patients 

with Type 2 Diabetes 

ZHIBO AN, SADIA ALI, COLLEEN ROGGE, FAY HAILES, CATHY BAILEY, BRENDA 

WENSTRUP, BRIANNE REEDY, MARZIEH SALEHI, DAVID A. D’ALESSIO, Cincinnati, OH 

The incretin effect is impaired in type 2 diabetes, and diabetic subjects have 

diminished responses to incretins. However, it is not clear whether the defects 

are specifi c for incretin stimulated insulin secretion or simply another aspect of 

generalized β-cell dysfunction. Correction of chronic hyperglycemia improves 

incretin action in animals. Further, normalization of hyperglycemia in diabetic 

patients improves the potentiation of glucose-stimulated insulin secretion 

by GLP-1 and GIP. The aim of this study was to determine whether glycemic 

control specifi cally improves the incretin effect in humans. Six type 2 diabetic 

subjects with moderate to poor control (age 55±3; BMI 34±2) were studied 

twice using a glucose clamp-OGTT protocol before and after 8 weeks of longacting 

insulin treatment titrated to a fasting glucose target of 6.0 mM, causing 

Hg A1C to reduce from 8.2±0.2 to 6.8±0.2 %. Following an overnight fast and 

basal blood draws (-20-0 min), glucose was given intravenously (IV) to raise 

blood glucose by ∼6.5 mM. At 90 min 75 g of glucose was given orally, and 

the IV glucose infusion was adjusted to maintain the blood glucose constant. 

The incretin effect was calculated by comparing plasma insulin levels before 

and after the oral glucose load. In the basal period, P1 (0-90 min) and P2 (90- 

270 min), the blood glucose levels were 8.6±0.6, 15.1±0.8, and 16.3±1.3 mM 

at visit 1; and 5.9±0.6, 13.2±0.8, and 14.3±1.0 mM at visit 2, respectively. 

The incremental insulin response to IV glucose (P1) increased from 0.10±0.06 

(visit 1) to 0.16±0.08 nM (visit 2). The response to oral glucose ingestion at 

fi xed hyperglycemia (P2) increased from 0.42±0.30 (visit 1) to 0.90±0.38 nM 

(visit 2). After 8 weeks of treatment, the average increase of the incremental 

insulin response to oral glucose was 8 fold greater than the increase to IV 

glucose (p=0.2), with 5 out of 6 subjects having a 4 fold or greater increase. 

Our data support the hypothesis that intensifi ed insulin treatment to improve 

glycemic control leads to a disproportionate improvement of insulin secretion 

in response to oral, compared to isoglycemic IV, glucose stimulation in patients 

with type 2 diabetes. 

Supported by: VA Merit Award to D.D. 



A484 

1789-P 

Involvement of Hypothalamic AMPK and Histamine on Olanzapine- 

Induced Glucose Intolerance in Mice 

MEGUMI ASATO, YOKO ISHIKAWA, HIROKO IKEDA, ATSUKO KAMEI, KENJI 

ONODERA, JUNZO KAMEI, Tokyo, Japan, Kanagawa, Japan 

Atypical antipsychotic drugs are well known to produce metabolic 

disturbance. Treatment with some atypical antipsychotic drugs such as 

clozapine or olanzapine induces the impairment of glucose metabolism, 

which increases the risk for developing metabolic side-effects, including 

weight gain, dyslipidemia, insulin resistance and hyperglycemia. We have 

already reported that central administration of olanzapine produces glucose 

intolerance. Recent study showed that clozapine activated adenosine 

5’-monophosphate-activated protein kinase (AMPK) through histamine H1 

receptors in the hypothalamus. AMPK is known to sense the energy status 

of cells and to regulate fuel availability. In central nervous system, AMPK 

modulates feeding and systemic energy homeostasis. Thus, it is possible 

that atypical antipsychotic drugs such as clozapine and olanzapine induces 

glucose intolerance by activating AMPK in the hypothalamus. Therefore, the 

present study was designed to investigate the involvement of histamine and 

AMPK on olanzapine-induced glucose intolerance. In the glucose CII test, 

both intraperitoneally (i.p.) or intracerebroventricually (i.c.v.) administration 

of olanzapine dose-dependently induced glucose intolerance as compared 

with the vehicle-treated group. The glucose intolerance induced by i.c.v. 

treatment with olanzapine was attenuated by i.p. pretreatment with 

histamine synthesis inhibitor, α-fl uoromethylhistidine (FMH). The glucose 

intolerance induced by i.c.v. treatment with olanzapine was also attenuated 

by i.c.v. pretreatment with an AMPK inhibitor, compound C. In addition, 

i.c.v. treatment with an AMPK activator, 5-aminoimidazole-4-carboxamide 

ribonucleoside (AICAR) also produced the same changes in serum glucose 

levels as olanzapine. In the western blot test, olanzapine increased the 

phosphorylation of AMPK in the hypothalamus, though total amount of 

AMPK was not affected. These results indicate that both histamine and 

AMPK in the hypothalamus are involved in the olanzapine-induced glucose 

intolerance. Furthermore, the present study suggests that olanzapine might 

activate hypothalamic AMPK via histaminergic system. 

1790-P 

Low Estradiol Concentrations in Males with Hypogonadotrophic 

Hypogonadism and Type 2 Diabetes 

SANDEEP DHINDSA, RICHARD FURLANETTO, MEHUL VORA, HUSAM GHANIM, 

PARESH DANDONA, Buffalo, NY, Chantilly, VA 

One-third of men with type 2 diabetes have hypogonadotrophic hypogonadism(HH). 

It has been suggested that HH in these men may be due to 

an increase in plasma estradiol(E 2 ) concentrations secondary to an increase 

in aromatase activity in the adipose tissue which leads to the suppression 

of hypothalamo-hypophyseal-gonadal(HHG) axis. We investigated the 

hypothesis that plasma E 2 concentrations are signifi cantly greater in type 2 

diabetic males with HH as compared to those without HH. Plasma estradiol, 

testosterone(T), LH and sex hormone binding globulin(SHBG) concentrations 

were measured in fasting blood samples of 236 men with type 2 diabetes 

(mean age: 56±12; range:23-83 years; mean BMI: 35±7;range:17-59kg/ 

m 2 ) attending a tertiary diabetes referral center. Total T was measured by 

liquid chromatography tandem mass spectrometry(LC-MS/MS). In 196 men, 

total E 2 was measured by an immunoassay. Free E 2 and T concentrations 

were calculated using total E 2 , T, albumin and SHBG. In 99 men, total E 2 

was measured by the more specifi c LC-MS/MS assay. Free E 2 and free T 

concentrations in these men were measured by tracer equilibrium dialysis. 

HH was defi ned as free T

1791-P 

Mechanism Underlying Metformin-Induced Secretion of Glucagon- 

Like Peptide-1 from the Intestinal L-Cell 

ANDREW J. MULHERIN, AMY H. OH, HELENA KIM, ANTHONY GRIECO, LINA M. 

LAUFFER, PATRICIA L. BRUBAKER, Toronto, ON, Canada 

The incretin hormone glucagon-like peptide-1 (GLP-1) is secreted by the 

intestinal L-cell in response to both nutrient and neural stimulation, leading 

to enhancement of glucose-dependent insulin secretion. GLP-1 is therefore 

a most attractive therapeutic approach for the treatment of Type 2 Diabetes 

Mellitus. The anti-diabetic drug, metformin, has previously been shown 

to increase circulating levels of GLP-1, although its mechanism of action 

is currently unknown. To elucidate this mechanism, GLP-1 secretion was 

measured in murine GLUTag, human NCI-H716, and rat FRIC L-cell cultures 

treated with metformin (5-2000µM) or AICAR (100-1000µM), activators of 

AMPK. Neither metformin nor AICAR directly stimulated GLP-1 secretion, 

despite a 1.7±0.2-fold increase in AMPK phosphorylation (P


Obesity 

POSTERS 

PEDF by adipocytes. The negative correlation between Vitamin D and PEDF is 

novel, and may be related to inverse correlations of Vitamin D and PEDF with 

metabolic risk factors. Further prospective studies are needed to determine 

the chronology and clinical importance of PEDF changes in diabetes. 

1795-P 

Potential Gastrointestinal Mechanisms for the Anti-Diabetic Effect 

of Metformin 

LIHONG CHEN, CHARI D. SMITH, YAPING LIU, MAGGIE S. MCINTYRE, DANA P. 

DANGER, JUDI A. MCNULTY, TYMISSHA D. JACKSON, SEAN A. ROSS, JEAN- 

LOUIS D. KLEIN, JAMES M. WAY, CHRISTOPHER C. NYSTROM, DALLAS K. CROOM, 

DONALD I. ANDERSON, ALAN J. CUNNINGHAM, ANDREW A. YOUNG, Research 

Triangle Park, NC 

Metformin is a fi rst-line oral treatment for type 2 diabetes mellitus. Its 

many reported mechanisms of action include increasing the sensitivity of 

liver, muscle, fat, and other tissues to insulin. Despite ample evidence for 

the effi cacy of orally-administered metformin, there are no clinical reports 

of an antidiabetic effect of parenterally-administered metformin. Its action 

may involve gastrointestinal (GI) targets where concentrations are high at 

effective doses (up to 2500 mg in humans). We evaluated its effects on the 

GI mechanisms, sodium-glucose co-transport (SGLT1) and apical sodiumdependent 

bile transport (ASBT), each of which exhibits antidiabetic effect 

in animal models. Uptake of labeled glucose and labeled taurocholate into 

cells respectively expressing human SGLT1 and human ASBT was assessed 

in vitro at different concentrations of metformin. It could dose-dependently 

and fully inhibit SGLT1 and ASBT, respectively, and was effective on those 

tranporters at concentrations estimated to prevail within the GI lumen at 

effective antidiabetic doses. Acute oral administration of metformin at 

doses that chronically correct diabetes in Zucker Diabetic Fatty (ZDF) rats 

(300mg/kg b.i.d.) signifi cantly decreased post-challenge glucose excursions 

in both ZDF and normal rats, consistent with an effect on glucose transport. 

Acute in vivo studies indicated that metformin could dose-dependently 

inhibit ileal bile salt reuptake, and increase fecal concentrations up to 3-fold. 

The chronic antidiabetic effects of metformin, manifest as dose-dependent 

reductions in non-fasting glucose and HbA1c in ZDF rats, were associated 

with an enhanced GLP-1 response in nutrient challenges. In summary, the 

glucose-lowering effi cacy of metformin in preclinical diabetic models could 

reside, at least partly, in the direct and/or indirect consequences of its 

luminal effects to inhibit SGLT1 and/or ASBT. 

1796-P 

Prolonged Initial Glucagon Response in Patients with Type 2 Diabetes 

Following a Mixed Meal 

MARJAN ALSSEMA, JOSINA M. RIJKELIJKHUIZEN, ELISABETH M. EEKHOFF, 

LEEN M. ‘T HART, GIEL NIJPELS, JACQUELINE M. DEKKER, Amsterdam, The Netherlands, 

Leiden, The Netherlands 

As a pancreatic hormone, glucagon promotes the conversion of liver 

glycogen into glucose when plasma glucose levels decrease. Inappropriate 

suppression of glucagon after oral glucose has been described for type 2 

diabetes patients. The effect of a physiological mixed meal stimulus on 

glucagon profi les as compared to oral glucose are largely unknown. 

In a population-based study, 21 persons with and 182 without type 2 

diabetes received a standardized mixed meal (75 g carbohydrates, 50 g 

fat and 24 g proteins) and an oral glucose tolerance test (75 g glucose) on 

separate occasions. Glucagon profi les up to t=120 minutes (oral glucose 

tolerance test) and t=240 minutes (mixed meal test) were measured by 

radioimmunoassay, after extraction. Time-dependent glucagon profi les 

were analysed by linear mixed models. 

Persons with diabetes had higher mean levels of glucagon in the fasting 

state (13.9 pmol/l versus 9.8 pmol/l, p

In multiple logistic regression analysis, serum A-FABP was an independent 

risk factor for CAD in women (OR=12.078, P=0.031). Serum A-FABP increased 

signifi cantly in multi-vessel diseased patients than in non-CAD subjects 

(P=0.011 in men, P=0.004 in women), and showed a positive correlation 

with CAI (r=0.134, P=0.032) after adjustment for age and gender. In addition, 

amino terminal pro-brain natriuretic peptide (NT-proBNP) was demonstrated 

to be positively and independently correlated with A-FABP in all subjects 

(β=0.075, P=0.014). 

Conclusions: Serum A-FABP is closely associated with the presence and 

severity of CAD in Chinese women. A-FABP levels are also associated with 

the circulating levels of NT-proBNP. 

Supported by: Chinese National 973 Project (2007CB914702) 

1799-P 

SIRT1-Mediated Regulation of FGF21 Expression in Lean Mice Probed 

with Small Molecule Activators and Genetic SIRT1 Deletion 

ANGELA M. COTE, MARC O. JOHNSON, KRISTINE STEARNS, MEGHAN L. DAVIS, 

DAVID J. GAGNE, MARIE YEAGER, JAMES L. ELLIS, VIPIN SURI, GEORGE P. 

VLASUK, Cambridge, MA 

The endocrine hormone Fibroblast Growth Factor 21 (FGF21) has broad 

metabolic actions including weight loss, increased insulin sensitivity 

and reduction of triglycerides in rodent models of obesity and metabolic 

dysfunction. FGF21 is induced in mouse liver by prolonged fasting, a 

ketogenic diet, and by agonists of peroxisome proliferator activated 

receptor alpha (PPARa). The protein deacetylase SIRT1 also modulates FGF21 

expression as hepatocytes defi cient in SIRT1 show reduced PPARa induced 

FGF21 expression while modest overexpression of SIRT1 increases FGF21 

expression in hepatocytes. However, the mechanism of SIRT1-mediated 

regulation of FGF21 secretion in mice has not been fully explored. 

To further understand the role of SIRT1 in mice, we analyzed the effect of 

genetic and pharmacological manipulation of SIRT1 activity on the induction 

of FGF21 by fasting as well as by two specifi c ligands of PPARa. A 24 hour fast 

increased FGF21 expression in liver as well as serum FGF21 levels by 8-10 fold. 

Oral administration of a specifi c small molecule SIRT1 activator during fasting 

further enhanced FGF21 serum levels by about two-fold. Administration of 

PPARa ligands WY14643 or GW7647 to ad lib fed mice also resulted in a dose 

dependent increase in hepatic FGF21 expression as well as serum FGF21 levels 

of up to 60 fold over vehicle treated ad lib fed mice. Concomitant administration 

of a sub-maximal dose of PPARa agonists WY14643 or GW7647 and a SIRT1 

activator to ad lib fed mice signifi cantly increased serum FGF21 levels beyond 

those induced by the PPARa agonist alone. 

We also analyzed the effects of fasting or PPARa agonists on FGF-21 

induction in SIRT1 conditional knockout mice. In contrast to the positive 

action of SIRT1 activator on FGF-21 induction, genetic deletion of SIRT1 

signifi cantly muted induction of FGF21 expression in liver as well as serum 

FGF-21 levels. Taken together these observations suggest a key role for 

SIRT1 in the regulation of FGF-21 levels. In addition, these observations 

suggest that regulation of FGF-21 levels may contribute to the metabolic 

benefi ts of genetic or pharmacological SIRT1 activation. 

1800-P 

Sitagliptin Suppresses Ghrelin Hormone in Patients with Diabetes 

BERHANE SEYOUM, ALEMU FITE, ABDUL B. ABOU-SAMRA, Detroit, MI 

Ghrelin is an appetite-stimulating hormone mainly produced by the stomach. 

Circulating levels of ghrelin concentration increase in fasting states and fall 

following meal. Sitagliptin is an orally available new class of anti-diabetic 

drug that inhibits dipeptidyl peptidase-4 (DPP-4) enzyme leading to 2-3 fold 

increase in the serum concentration of endogenous glucagon-like peptide-1 

(GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). This study 

was performed to determine the effects of sitagliptin on circulating levels 

of ghrelin. Control subjects (N=15) and 46 diabetic patients were recruited 

for the study. The diabetic patients were treated with sitagliptin (N=15), 




A487 

metformin (N=16) or combination of sitagliptin and metformin (N=15) for one 

week. Serum concentrations of total and active ghrelin were determined 

in all subjects immediately before and 2 hours after meal challenge. Same 

testes were repeated among patients with diabetes after receiving drug 

therapy for a week. Results demonstrated that postprandial active ghrelin 

was more signifi cantly suppressed in patients with diabetes than in nondiabetic 

controls. Maximum suppression was seen after patients were put 

on treatment and after meal challenge. Active ghrelin was more signifi cantly 

decreased than total ghrelin in diabetic patients (by 36%, p0.05) whereas active ghrelin decreased 

from 140±26 pg/ml to 85±12 pg/ml (p


Obesity 

POSTERS 

but glucagon remains inappropriately elevated during eu- and hyperglycemia. 

Normalization of glucagon requires reduction of FIG accompanied by only minor 

FRG suppression. Table 1 summarizes the impact of ACI on the fold increase in 

glucagon in response to hypoglycemia assuming 0% to 100% reduction in FIG 

(by rows) and FRG (by columns). Values >7 (bold) indicate restoration to levels 

similar to the normal pancreas (vs. 1.6 in insulin defi ciency). 

FIG↓/FRG→ 0% 20% 40% 60% 80% 100% 

0% 1.6 1.5 1.4 1.4 1.3 1 

20% 4.3 2.8 2 1.6 1.5 1 

40% 8.6 7.5 6 3 1.9 1 

60% 13.3 11.6 9.8 7.7 3.1 1 

80% 18.9 17.3 15.6 13.1 7.2 1 

100% 28.8 28.8 28.8 27.2 28.3 

In conclusion, repair of defective GCR is possible by ACI that suppress 

preferentially the basal rather than the pulsatile glucagon release. Since 

deconvolution of hormone time series can distinguish between these two 

secretory components our results suggest a hypothesis to test experimentally 

that ACI with mostly basal suppression may be used as a protection against 

hypoglycemia in insulin defi ciency. 

Supported by: NIDDK grant R01 DK082805 

1803-P 

The Relationships of Pancreatic Polypeptide with Aging, Obesity 

and Diabetes 

ZHIKE CHEN, ZHUO LIU, CHEE W. CHIA, OLGA D. CARLSON, JOSEPHINE M. EGAN, 

Baltimore, MD 

Insulin, glucagon, pancreatic polypeptide (PP), somatostatin and ghrelin 

are secreted from fi ve distinct cells in islets of Langerhans. While the 

regulation of insulin secretion and its dysregulation in type 2 diabetes 

have been extensively investigated, the other four hormones are often over 

shadowed. Plasma glucagon levels are elevated in type 2 diabetes and are 

implicated in the pathogenesis of the elevated fasting glucose levels. PP 

has been largely ignored. We hypothesized that PP secretion might also be 

dysregulated in type 2 diabetes and insulin resistance states. The purpose 

of this study was to investigate the relationships between PP plasma levels 

with aging, obesity and diabetes in the fasting state and after oral glucose 

tolerance test (OGTT). 

Using data collected from the Baltimore Longitudinal Study of Aging 

(BLSA), we found that in non-diabetic healthy subjects the plasma PP levels 

were signifi cantly elevated in aged subjects (80-90 years old) compared with 

young subjects (30-40 years old) in the fasting state (50.2±7.6 vs. 30.6±5.0 

pM, p

estrictive and GI bypass models, respectively. By using SG and RYGB mouse 

models, we tested whether the effects of bariatric surgery in the treatment 

of obesity and insulin resistance are associated with the regulation of 

infl ammation in visceral adipose tissue (VAT) and liver in DIO mice. SG 

induced weight loss, reduced fat mass and improved glucose tolerance in 

glucose tolerance tests within 4 wks, but the effects were not completely 

sustained. However, RYGB resulted in a sustained prevention of weight gain 

and improved liver steatosis and insulin resistance through 12 wks. Both RYGB 

and SG inhibited CD11b macrophages and Th1 (CD3 + /IFNg + ) subsets in liver by 

-60% and -42%, respectively, as well as in VAT (-85% and -64%, respectively) 

and enhanced Th2 (CD3 + /IL-10 + ) subsets in liver (+100%), but not in VAT at 7 

days post-surgery. RYGB, but not SG, suppressed phosphorylation of IRS- 

1/Ser 307 , inhibited (-47%) M1 (CD11c) macrophage polarization, promoted 

(+78.5%) M2 (CD206) macrophage polarization and enhanced (+100%) CD3 + / 

FoxP3 + regulatory T cells in the liver. RYGB, but not SG, reduced circulating 

insulin (-85%), leptin (-45%) and IL-6 (-64%) by one week post-surgery. Our 

study suggests that SG induces weight loss and improves glucose tolerance in 

the short term; however, RYGB persistently improves insulin resistance. Early 

positive effects of RYGB and SG in the improvements of insulin resistance 

appear to be modulated by the inhibition of infl ammation in liver and VAT. 

RYGB specifi cally inhibits infl ammatory M1 macrophages, enhances M2 

macrophages and promotes regulatory T cell subsets in the liver. 

Supported by: NIH grants to D.W. and N.A., JDRF grant to D.Y. 

& 1807-P 

Depletion of α/β T Cells Attenuates Obesity-Associated Infl ammation 

and Metabolic Abnormalities 

ILVIRA M. KHAN, XIAOYUAN PERRARD, JERRY PERRARD, AMIR MANSOORI, C. 

WAYNE SMITH, HUAIZHU WU, CHRISTIE M. BALLANTYNE, Houston, TX 

Recent investigations have linked obesity with low-grade chronic 

infl ammation in adipose tissue (AT), which causes AT dysfunction, thus 

contributing to obesity associated metabolic abnormalities. Diet-induced 

obesity is correlated with accumulation of “classically activated” (M1) 

macrophages in AT, which secret proinfl ammatory cytokines and impair 

insulin signaling and lipid storage in adipocytes. In addition, obesity alters T 

cell numbers in AT. However, the role of T cells in metabolic complications 

of obesity is less defi ned. Using mouse model of diet-induced obesity, we 

found that both αβ and γδ T cells reside in perigonadal AT of lean mice. 

However, only αβ T cells were signifi cantly increased in AT of obese mice 

(25.05 + 4.38%) compared with lean controls (8.89 + 3.60%, n=7/group, 

P


Obesity 

POSTERS 

versus controls, despite similar intake), which may be attributed in part to 

an increased locomotor activity (p

the ASP1941 10 mg/kg group (P


Obesity 

POSTERS 

indicates that MβKO mice have increased adiposity. Signifi cantly elevated 

serum leptin levels were observed in MβKO mice, despite the fact that 

their hematological parameters, serum insulin and glucagon were within 

normal ranges. Leptin-suppressed food intake and body weight reduction 

in MβKO mice were signifi cantly less than WT mice, which further suggest 

that knocking out C/EBPβ in macrophages impairs leptin sensitivity in mice. 

The expression of key lipogenic genes was signifi cantly increased in livers 

and white fats of MβKO mice. On the other hand, the levels of F4/80 and 

other macrophage markers were remarkably decreased in white fat, liver 

and skeletal muscle of chow or high fat diet-fed MβKO mice. Using bone 

marrow-derived macrophages, we found the expression of alternative 

activated macrophage markers were signifi cantly decreased in C/EBPβ 

defi cient macrophages at basal and after IL-4 induction. These results 

indicate that 1) C/EBPβ plays an important role in macrophage activation 

and tissue infi ltration; 2) residential macrophages may actively regulate 

energy metabolism. 

Supported by: NIDDK ADA-Funded Research 

& 1819-P 

Sterol Regulatory Element Binding Protein-1c (SREBP-1c) Is Different 

ly Regulated by TNF-Related Apoptosis Inducing Ligand (TRAIL) 

in High-Fat Diet Induced Obese Mice 

SO-YOUNG PARK, MI-KYOUNG PARK, YING HAN, SU KYUNG PARK, DUK KYU 

KIM, HYE-JEONG LEE, Busan, Republic of Korea 

TRAIL is a member of TNF family of cytokines, which exist either as type II 

membrane or as a soluble protein. Although the well-characterized activity of 

TRAIL is represented by its anti-cancer activity, little is known regarding the 

effects of TRAIL on metabolic pathways. To address this point, we studied 

in vivo effects of TRAIL in high-fat diet induced obese mice. SREBP-1c is a 

transcription factor which is synthesized as a precursor in the membranes of 

the endoplasmic reticulum and which requires post-translational modifi cation 

to yield its transcriptionally active nuclear form. SREBP-1c induces the 

expression of a family of genes involved in glucose utilization and fatty 

acid synthesis. C57BL/6 Mice were classifi ed into two groups and fed with 

the normal (ND) and high-fat diet (HFD) for up to 22 weeks. All mice of the 

high-fat diet group developed obesity and type 2 diabetes. Seven days after 

adenoviral-mediated hTRAIL (ad.hTRAIL) and control GFP (ad.GFP) delivery 

to each groups, mice were killed and samples were collected and analyzed. 

The expression of ad.hTRAIL was determined by western blotting with liver 

tissue. Overexpression of ad.hTRAIL signifi cantly decreased accumulation of 

hepatic lipid and plasma lipid profi les in HFD mice. The expression of hepatic 

SREBP-1c was increased by ad.hTRAIL in ND mice, compared with ad.GFP. 

The expression of SREBP-1c was increased in HFD mice, compared with 

ND mice as expected by high lipogenic activity. However, the expression of 

SREBP-1c was decreased by ad.hTRAIL in HFD mice, compared with ad.GFP 

in HFD mice. The target genes of SREBP-1c, fatty acid synthase, acetyl CoA 

carboxylase, and stearoyl CoA desaturase 1 were altered signifi cantly by 

the changes of the expression of SREBP-1c. Taken together, TRAIL alters 

the expression of lipogenic genes through SREBP-1c and the expression 

of SREBP-1c is differently regulated by TRAIL in response to high-fat diet. 

These fi ndings suggest that TRAIL might have a novel function for lipid 

metabolism via SREBP-1c related mechanism, modulating lipogenesis in 

lipid-rich environment and improving hepatic steatosis. 

& 1820-P 

Roux-en-Y Gastric Bypass Prevents the Progression to Diabetes in 

Obese Mice 

DENG-PING YIN, QIANG GAO, LIAN-LI MA, PHILLIP E. WILLIAMS, OWEN MC- 

GUINNESS, ALVIN C. POWERS, DAVID H. WASSERMAN, NAJI N. ABUMRAD, 

Nashville, TN 

Obesity is associated with insulin resistance and increased risk of 

developing type 2 diabetes (T2D). C57BLKS-db/db (BKS-db) mice, a model of 

insulin resistance, develop diabetes with aging. We tested whether Rouxen-Y 

gastric bypass (RYGB), an effective therapy for morbid obesity and T2D 

in humans, can prevent disease progression in young pre-diabetic (6-7 wks 

old, n=6) and/or reverse diabetes in older diabetic BKS-db mice (14-15 wks 

old, n=4), as well as in wild-type STZ-induced diabetic mice (FVB) expressing 

luciferase under the control of the mouse insulin promoter (MIP-Luc, n=6). 

All BKS-db mice that underwent RYGB were compared to pair-fed BKSdb 

mice that underwent a sham surgery (n=4/group). In the young BKS-db 

mice, RYGB and pair-fed sham-treated groups experienced similar weight 

loss and gain and changes in fat mass throughout the fi rst eight wks. RYGB 

in young BKS-db mice maintained normoglycemia (blood glucose levels < 

200mg/dl) and improved glucose tolerance during intraperitoneal glucose 


OBESITY—ANIMAL 

CATEGORY 

A492 

tolerance tests (area under curve) up to 8 wks after surgery (compared 

with pair-fed sham-treated mice, 993±189 vs 1856±169 at 1 wk, p

saline-treated group fed ad lib. CTL ASO did not change BW or BF versus 

saline-CR. However, R4 ASO further lowered BW by 7-11% and BF by 23- 

25%. In addition, R4 ASO reduced epididymal fat pad wt by 16-22% and 

peri-renal fat pad wt by 29-33% without effect on lean mass. As expected, 

CR signifi cantly reduced whole body VO 2 . In contrast, R4 ASO prevented the 

decline in VO 2 . In separate studies, R4 ASO Rx of lean C57BL/J and CD-1 

mice for 3 mo resulted in > 75% reduction in liver FGFR4 mRNA and was well 

tolerated without any overt side effects, indicating lack of mechanism based 

toxicity with chronic reduction of FGFR4. To further explore mechanism of 

action, the effect of R4 ASO Rx on FGF15 levels was determined. Four-wk 

Rx of DIO mice reduced liver FGFR4 mRNA by > 75%, increased ileum FGF15 

mRNA by > 5-fold and plasma FGF15 protein levels by > 2-fold. These rodent 

fi ndings were confi rmed in Cynomolgus monkeys, where Rx with R4 ASO for 3 

mo reduced liver FGFR4 mRNA by 70% and caused > 5-fold increase of ileum 

FGF19 mRNA and 2-fold increase of plasma FGF19 protein levels without any 

overt toxicities. These data provide further support that peripheral inhibition 

of FGFR4 with antisense drugs is an attractive therapeutic approach for 

obesity. 

1823-P 

Caloric Restriction Reverses Elevated ER Stress and Oxidative 

Stress in Liver of ob/ob Mice 

ATSUYUKI TSUTSUMI, HIROYUKI MOTOSHIMA, SHUJI KAWASAKI, SATOKO 

HANATANI, MOTOYUKI IGATA, TATSUYA KONDO, TAKESHI MATSUMURA, KAKU 

TSURUZOE, TAKESHI NISHIKAWA, EIICHI ARAKI, Kumamoto, Japan 

Endoplasmic reticulum (ER) stress and oxidative stress have been 

proposedto play a crucial role in the development of insulin resistance 

and diabetes in obesity. Although caloric restriction (CR) improves various 

obesity-related disorders, the effects of CR on ER stress and oxidative stress 

in obesity have not been elucidated. 

To investigate how CR affects ER stress, oxidative stress and insulin 

signaling in obesity, a leptin-defi cient ob/ob mice under CR (ob-CR) or ad 

libitum (AL)–feeding (ob-AL) for four weeks were compared on daily foodintake, 

body weight (BW), glucose metabolism, ER stress, oxidative stress 

and insulin signaling. To reduce BW to the level of lean littermates fed AL 

(lean-AL), the ob-CR were given reduced chow (2.0 g/day) to a level which 

previously reported to extend lifespan of ob/ob mice. Glucose and insulin 

tolerance, markers for ER stress and oxidative stress (protein nitrotyrosine), 

and insulin signaling were investigated in these animals. 

CR reduced BW in ob-CR, resulted in comparable BWs between ob-CR 

and lean-AL after 2 weeks, both of which became smaller than ob-AL in BW. 

The ob-CR showed improved glucose tolerance and hepatic insulin action 

compared with ob-AL. Signifi cant increases in ER stress markers (such as 

phosphorylation of PERK and eIF2α, and expression of ATF4 and GRP78 

mRNA) and in protein nitrotyrosine were observed in liver and epididymal 

fat from ob-AL compared with those from lean-AL. CR signifi cantly reduced 

all of these markers in ob/ob mice. CR also signifi cantly reduced both serinephosphorylation 

of IRS-1 and JNK phosphorylation in liver of ob/ob mice. 

Reduction in ER stress by CR tended to be stronger than that obtained by 

treatment with 4-phenyl butyric acid, which is known to reduce ER stress 

in vivo. 

In conclusion, four weeks of CR effectively reduced ER stress and oxidative 

stress, and improved insulin action via suppression of JNK-mediated IRS-1 

phosphorylation in liver of ob/ob mice. 

1824-P 

Catechol-o-Methyltransferase Defi ciency Is Associated with Ab normal 

Adiposity and Glucose Intolerance: Implication for a New Mechanism 

of Metabolic Syndrome 

MEGUMI KANASAKI, KEIZO KANASAKI, DAISUKE KOYA, Uchinada, Japan 

Catechol-o-methyltransferase (COMT), an enzyme linked with several 

neuronal/psychological diseases, metabolizes a group of catechols including 

catecholamines and catechol estrogens. 2-methoxyestradiol, one of the 

metabolite of catechol estrogens via COMT, exerts as an anti-angiogenesis 

and a vascular protective factor via inhibition of hypoxia-inducible factor 

(HIF)-1α. Several single-nucleotide polymorphisms (SNPs) in the human 

COMT gene have been observed and some of which may result in the 

depletion of catalytic activity, such as the well-known functional Val158Met 

isoform. These COMT SNPs have been associated with hypertension, 

preeclampsia, obesity, increased fat mass and obesity in diabetes. However, 

the contribution of COMT suppression in the onset of metabolic diseases 

has not been determined. Using COMT-inhibitor (Ro 41-0960), here we 

show that COMT-inhibitor accelerate adiposity with the impaired glucose 

tolerance in mice fed a high-fat diet. A 2-week high-fat diet (60% fat in total 




CATEGORY 

A493 

energy) induced impaired glucose tolerance when analyzed by the intraperitoneal 

glucose tolerance test in mice. COMT-inhibitor treatment for last 

1 week displayed the exacerbation of glucose tolerance defects and insulin 

resistance in mice with a high-fat diet. Such impaired glucose tolerance 

was associated with the hepatosteatosis, decreased liver glycogen level 

associated with HIF-1α and macrophage accumulation in epididymal fat. A 

signifi cant angiogenesis in the mesenteric fat was also observed in highfat 

fed mice treated with COMT-inhibitor. These abnormal adiposity and 

glucose tolerance defect with the abnormal angiogenesis in COMT-inhibitor 

treated mice were ameliorated by the 2-ME treatment. 2-ME suppressed 

HIF-1α and macrophage accumulation in the epididymal fat. These results 

indicated that 2-ME could be the potential target drug for the treatment of 

metabolic defects via inhibition of abnormal adipogenesis, angiogenesis and 

infl ammation especially in low COMT genotype population. 

Supported by: Kanae Foundation (K.K.) 

1825-P 

Cathepsin K, L Expression in Adipocytes Is Upregulated by Palmitate 

Via TNFα and IL-6 

JEONG SEON YOO, YOUNGMI LEE, EUN HAE LEE, JIWOON KIM, SHINYOUNG 

LEE, JANG-HAN JUNG, JI SUN NAM, SHIN AE KANG, TAE-WOONG NOH, MIN 

HO CHO, JONG SUK PARK, KYUNG WOOK KIM, JAE-WOO KIM, CHUL WOO AHN, 

BONG SOO CHA, EUN JIG LEE, SUNG KIL LIM, KYUNG RAE KIM, HYUN CHUL LEE, 

Seoul, Republic of Korea 

Aim: Cathepsin family is lysosomal cysteine protease. It is recently 

reported that cathepsin K, L, S control adipogenesis and relate to acute 

coronary syndrome. However, it is not clear whether cathepsins expression 

can be affected by saturated fatty acid which has a critical role in metabolic 

disease. We examined the hypothesis that palmitate upregulates cathepsin 

K, L, S expression in obese adipose tissue and infi ltrating macrophages have 

crucial role in this process via infl ammatory cytokines. 

Methods and Results: 3T3-L1 cells were fully differentiated to mature 

adipocytes for 8 to 10 days and treated with palmitate, lipopolysaccharide 

(LPS) which is ligand of TLR4 (toll-like receptor 4). Real-time PCR revealed that 

not cathepsin S but cathepsin K, L expression is upregulated by palmitate as 

dose- and time-dependent manner. But there was no additional effect when 

we use palmitate-treated RAW264.7 cells’ media instead of direct treatment 

to 3T3-L1 cells. Cathepsin K, L is upregulated after treatment of TNFa or IL-6 

like palmitate. Six-week-old C3H/HeN mice with normal TLR4 and C3H/HeJ 

mice with TLR4 gene’s mutation got LPS injection ip (1 mg/kg) or were fed 

with high fat diet (60% fat of total calories) for 13 weeks and sacrifi ce to 

harvest epididymal fat. Cathepsin K, L, S expression increased in both kinds 

of mice fed high fat diet compared with chow diet. But we cannot fi nd similar 

pattern at mice which got LPS ip injection. 

Conclusions: We concluded that cathepsin K, L expression in fat tissue are 

upregulated by saturated fatty acid via infl ammatory cytokines, not TLR4, 

thereby might have a critical role in developing acute coronary syndrome 

of metabolic syndrome and there are no additional effects of infi ltrating 

macrophages. 

1826-P 

Comparison of Genetic Factors Controlling Abdominal Fat Distribution 

and Glucose Metabolism in A/J and SM/J Mice 

MISATO KOBAYASHI, TAMIO OHNO, MASAKO KUGA, MASAHIKO NISHIMURA, 

ATSUSHI MURAI, FUMIHIKO HORIO, Nagoya, Japan 

Obesity is a major risk factor for insulin resistance, type 2 diabetes, 

dys lipidemia, cardiovascular disease, fatty liver and stroke. However, 

each abdominal fat depot, such as mesenteric or epididymal, differently 

contributes to the development of insulin resistance. Previously, we 

mapped a major quantitative trait locus (QTL, T2dm2sa) for impaired glucose 

tolerance on chromosome (Chr.) 2 and revealed that the chromosomal region 

near T2dm2sa on Chr. 2 had affective genes on not only glucose tolerance, 

but also the accumulation of body fat, using SM.A-T2dm2sa congenic 

mice. SM.A-T2dm2sa mice possess the A/J-allele T2dm2sa region in SM/J 

mice. To identify the genetic regions that contribute to fat accumulation in 

epididymal and mesenteric fat depots and to examine whether or not the 

genetic regions that affect glucose metabolism and body fat distribution 

are coincident, we performed QTL analyses using (A/J×SM.A-T2dm2sa)F2 

intercross mice. SM.A- T2dm2sa mice shows lower glucose tolerance and 

higher epididymal fat weight compared with A/J mice. 

Signifi cant QTLs for mesenteric fat were detected on Chrs. 5 (Mfatq3sa, 

LOD: 3.5), 7 (Mfatq1sa, LOD: 7.7) and 16 (Mfatq2sa, LOD: 5.0). Signifi cant 

QTLs for epididymal fat were detected on Chrs. 10 (Efatq2sa, LOD: 3.8) and 



Obesity 

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Obesity 

POSTERS 

12 (Efatq1sa, LOD: 4.5). Thus, QTLs for mesenteric and epididymal fat depot 

were mapped on the different chromosomal regions. The A/J allele on Chrs. 

7, 10 and 16 contributed to increase mesenteric or epididymal fat weights, 

oppositely the SM/J allele on Chrs. 5 and 12 contributed to increase these 

fat weights. This suggests that the fat accumulations in individual fat depots 

are controlled by distinct genomic regions. In addition, the affected allele 

exists in both of the parental strains. 

Signifi cant QTLs for impaired glucose tolerance and blood glucose 

concentration have been detected on Chrs. 3, 6, 11 and 18. Comparison of 

these QTLs for abdominal fat distribution with those for glucose metabolism 

revealed that the genetic factors controlling fat accumulation in adipose 

tissue do not coincide with those controlling glucose tolerance and blood 

glucose concentration in (A/J×SM.A-T2dm2sa) F2 mice. 

1827-P 

Effect of Sucrose vs. Fructose-and-Glucose on the Body Mass of the 

Rat 

GRZEGORZ WYSTRYCHOWSKI, WLADYSLAW GRZESZCZAK, EWA OBUCHO- 

WICZ, ANTONI WYSTRYCHOWSKI, Zabrze, Poland, Katowice, Poland 

U.S. boost in obesity coincided with high-fructose corn syrup (HFCS, 

mixture of 55% fructose (F) and 42-45% glucose (G)) domination as sweetener 

in soft drink industry. Obesogenic effect of HFCS has been claimed with 

little scientifi c evidence and attributed to syrup-specifi c compounds like 

dicarbonyls or higher F content as compared with sucrose (S). We aimed to 

verify it by assessing impact of S vs. HFCS-like mixture of F&G on weight and 

basic metabolic markers in rat model. 

24 M 6-w.o. S-W rats were assigned to 9 weeks of unlimited drinking of 

5% S solution (8 rats, S group), 5% solution of 55% F & 45% G (8 rats, F&G) 

or tap water (8 rats, W). Chow was provided ad libitum. Metabolic markers at 

study completion, weight changes, and fl uid, chow and energy intakes were 

compared between groups. 

F&G and W gained more weight than S (table). There was trend for fl uid, 

chow, and corresponding caloric intakes to be greater in F&G than in S. 

Total energy intake in W tended to be higher than in S, while lower than in 

F&G. Serum cholesterol was higher in F&G and S. Triglycerides tended to be 

increased in F&G. Uric acid and G levels did not differ (not shown). 

S F&G W 

Baseline weight [g] 184±19 175±7 174±15 

Weight gain/day [g] 3.9±0.4 4.5±0.4** 4.7±0.6 ## 

Fluid intake/day [ml] 80.5±9.2 89.2±15.0* 51.1±4.2 ##$$ 

Chow intake/day [g] 21.1±1.1 22.9±1.0* 27.5±1.0 ##$$ 

Energy from fl uid/day [kJ] 65.2±7.5 69.4±11.7* 0 ##$$ 

Energy from chow/day [kJ] 253.7±13.1 274.7±12.3* 330.6±11.7 ##$$ 

Total energy intake/day [kJ] 318.9±5.7 344.1±23.9* 330.6±11.7 #$ 

Serum cholesterol [mmol/l] 1.17±0.17 1.23±0.15 0.93±0.13 ##$$ 

Serum triglycerides [mmol/l] 0.72±0.25 0.99±0.39 0.66±0.24 $ 

*P

were improved by fenofi brate and pioglitazone. High fat diet induced TG 

accumulation in skeletal muscle was also decreased 20% by fenofi brate 

and 24% by pioglitazone. ATGL protein expression was respectively reduced 

40% and 18% in adipose tissue and skeletal muscle of HFD rats and was 

increased after pioglitazone treatment in these tissues. ATGL was increased 

in skeletal muscle but has no change in adipose tissue by fenofi brate. 

Fenofi brate and pioglitazone treatment improved insulin sensitivity and 

increased ATGL protein and coincide with decreased TG content in skeletal 

muscle. These fi ndings suggest that fenofi brate and pioglitazone ameliorate 

insulin resistance through decreased ectopic TG content mediated by 

increased ATGL in mainly tissues. 

Supported by: National Natural Science Foundation of China Project (30670989) 

1831-P 

Fenofi brate Involves in CCK-Induced Reduction of Food Intake in 

Rats 

YING HAN, MI-KYOUNG PARK, SO-YOUNG PARK, SU KYUNG PARK, DUK KYU 

KIM, HYE-JEONG LEE, Busan, Republic of Korea 

Fenofi brate has been reported to decrease food intake independent of 

leptin in rodents. We hypothesized that fenofi brate might decrease food 

intake via cholecystokinin (CCK) dependent pathway. Otsuka Long-Evans 

Tokushima fatty (OLETF) rats, which genetically lack CCK receptor as a result 

of mutation, were divided into OLETF-fenofi brate group (n=5) and OLETFcontrol 

group (n=5). OLETF-fenofi brate group was fed with standard rat chow 

and fenofi brate (30mg/kg/day) and OLETF-control group was only fed with 

standard rat chow for the same period as control. Long-Evans Tokushima 

Otsuka (LETO) rats which have normal CCK receptor, as control groups to 

OLETF rats, were also divided into LETO-fenofi brate group (30mg/kg/day) 

(n=5) and LETO-control group (n=5). After 11 weeks of fenofi brate treatment, 

food intakes of the fenofi brate group were signifi cantly decreased than those 

of the control group in the LETO rats (P< 0.05), but there was no signifi cant 

difference in the OLETF rats. The levels of blood β-ketone, plasma leptin, 

and fasting blood sugar (FBS) did not show signifi cant difference between 

the fenofi brate and control groups neither in the LETO rats nor in the OLETF 

rats. To investigate the expression of CCK by fenofi brate, the protein level 

of CCK were analyzed with small intestinal tissue of the rats. Fenofi brate 

treatment groups showed that the protein levels of PPARa and CCK were 

signifi cantly increased in the small intestine than control groups both in the 

LETO and OLETF rats. To examine the expression of CCK by fenofi brate, the 

Caco-2 cells, intestinal epithelial cell line, were cultured in the presence of 

fenofi brate for 24 hours. Fenofi brate treatment signifi cantly increased the 

protein expressions of PPARα and CCK in the Caco-2 cells. In conclusion, 

fenofi brate treatment decreased food intake in the LETO rats which have 

normal CCK receptor, but not in the OLETF rats which lack CCK receptor. The 

possible anorexigenic mechanism by fenofi brate might be up-regulation of 

CCK in the small intestine. 

1832-P 

High Free Fatty Acids Level Related with Microalbuminuria in Obese 

Rats 

XIAO DONG SUN, YE RONG YU, LI NA HAN, BEN WANG, Chengdu, China 

It is known that obesity is associated with microalbuminuria (MAU), which 

is not only an early manifestation of kidney damage, but also an independent 

risk factor for ischemic cardiovascular disease. High free fatty acids (FFAs) 

level is a common feature of obesity and can cause impaired endotheliumdependent 

vasodilatation (EDV). We hypothesized that increased release 

of FFAs from excessive visceral fat accumulation is related to increased 

urine albumin excretion and reduced circulating FFAs level by fenofi brate 

has renal protective effects in MAU by improving endothelial dysfunction 

in diet-induced obese rats. Male Wistar rats 4 weeks in age were 

randomly divided into three groups. The rats in the control group, obesity 

group and fenofi brate group were fed with normal diet, high-fat diet, and 

high-fat diet plus fenofi brate (100mg/kg/d), respectively. At the end of 24 

weeks, body weight increased 32.8% in obese rats compared to control 

group. The serum FFAs and TG levels increased signifi cantly in obese rats. 

Fenofi brate intervention decreased serum FFAs and TG levels by 43.4% and 

48% respectively, accompanied by reduced ratio of perirenal fat to body 

weight (PF/BW) and visceral fat to body weight(VF/BW). Urinary albumin/ 

creatinine ratio (ACR) increased in obese rats (56.09±22.64 VS 19.52±7.30 

mg/g, P


Obesity 

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reported in humans. Thus, fructose-fed rhesus monkeys represent a novel 

nonhuman primate model useful for investigating the pathophysiology, 

prevention, and treatment of the metabolic and infl ammatory changes that 

lead to an increased risk for type 2 diabetes and atherosclerotic cardiovascular 

disease. 

Supported by: NIH Grants R21 AT250099 and R21 AT003645 


1835-P 

Lack of Inteferon-γ Results in Reduced Body Weight and Better 

Glucose Tolerance 

NICOLE WONG, BARBARA C. FAM, GITTA R. CEMPAKO, GREGORY R. STEINBERG, 

THOMAS T.W. KAY, JOSEPH PROIETTO, SOFIANOS ANDRIKOPOULOS, Heidelberg 

Heights, Australia, Hamilton, ON, Canada, Fitzroy, Australia 

Obesity is a chronic low-grade infl ammatory disease that is caused by 

increased energy intake and reduced energy expenditure. Studies using 

animal models with deletion of infl ammatory cytokines have produced 

confl icting results with some showing increased weight gain and others 

showing no effect or even reduced body weights. Clearly more work is 

necessary to understand the role of cytokines on body weight control. 

Interferon-gamma (IFNγ) is a cytokine derived from T-cells that plays an 

important role in the innate and adaptive immune response particularly to 

viral infections. Emerging evidence suggests that increases in the T-cell 

population in adipose tissue may contribute to obesity and the associated 

metabolic syndrome. Recent studies have shown that IFNγ was increased 

with obesity in both human and rodent models while defi ciency of IFNγ 

resulted in better glucose tolerance in mice. However, how IFNγ affects 

glucose tolerance is not known. The aim of this study was to assess the 

effects of IFNγ on both body weight and glucose metabolism using a global 

knockout mouse model. To do this, male IFNγ -/- and wild-type C57BL/6 mice 

were monitored for 20 weeks for body weight, food intake and physical 

activity on a standard rodent chow diet (3.3 kcal/g). At the end of the study, 

IPGTT, ITT and hyperinsulinaemic/euglycaemic clamps were performed. 

Expression levels of arcuate nucleus NPY, AgRP and POMC mRNA as well 

as circulating leptin levels were also determined. The results show that 

IFNγ -/- mice had reduced weight gain associated with decreased food intake 

and increased physical activity. NPY and AgRP mRNA expression was 

reduced, while POMC mRNA expression was increased as were plasma 

leptin levels. This led to improved glucose tolerance and insulin sensitivity 

due to greater suppression of endogenous glucose output, associated with 

decreased hepatic glucose-6-phosphatase activity. We conclude that global 

deletion of IFNγ in mice resulted in reduced body weight associated with 

negative energy balance, glucose tolerance and improved hepatic insulin 

sensitivity. Our fi ndings demonstrate that IFNγ plays a critical role in the 

regulation of body weight and subsequently glucose metabolism. 

1836-P 

Neuronal Rictor Deletion Leads to Obesity, Growth Defects and 

Glucose Intolerance 

HEIDI E. KOCALIS, RICHARD L. PRINTZ, KEVIN D. NISWENDER, Nashville, TN 

Insulin has pleiotropic effects in the central nervous system (CNS), and 

neuronal resistance to insulin is implicated in the pathogenesis of obesity 

and dysglycemia. Akt is a down-stream mediator of insulin action on growth 

and metabolism. Akt is activated via phosphorylation at serine473 by the 

rapamycin-insensitive companion of TOR (Rictor) containing complex, 

mTORC2, in addition to PDK1 phosphorylation at threonine308. The role of 

mTORC2 signaling targets in CNS regulation of energy homeostasis is not 

well established. Given the role of Akt in energy and glucose homeostasis, 

and the role of mTORC2 in Akt activation, we hypothesized that neuronal 

Rictor gene deletion would lead to obesity and glucose intolerance. 

Cre-Lox P mediated deletion in neurons (Nestin-Cre) lead to gene dose 

dependant reductions in Rictor mRNA, and signifi cantly reduced serine473 

phosphorylation in null mice. Null mice displayed a profound growth defect 

that resolved by 12 weeks of age, after which, body weight and length were 

similar to controls. After 20 weeks of chow diet feeding, het mice gained 

68% (11.1±0.6g, P

1839-P 

Omega-3 Fatty Acids Protect Against Glucose Intolerance and 

Attenuate Ceramide Accumulation with High Fat Diet 

IAN LANZA, AGNIESZKA ZABIELSKI, PIOTR ZABIELSKI, DANIEL JAKAITIS, 

BUSHRA ALI, K. SREEKUMARAN NAIR, Rochester, MN 

Lipotoxicity is implicated as a cause of insulin resistance. Accumulation 

of lipid metabolites (e.g., ceramides) may interfere with insulin signaling in 

muscle. Dietary omega-3 fatty acids (n3PUFA) are reported to protect against 

insulin resistance with high-fat diets. We determined the impact of n3PUFA 

on the content and composition of lipid metabolites in skeletal muscle. Mice 

were fed for 10wks with normal fat diet (NF, 10% fat), high fat diet (HF, 60% 

fat), or high fat diet + fi sh oil (HF+N3, 60% fat with 3.4% kcals from n3PUFA). 

Oral glucose tolerance tests (OGTT) were performed at baseline and 10wks. 

Muscle tissue was homogenized and separated into sarcoplasmic, myofi brillar, 

and mitochondrial fractions. Ceramides and long chain acyl coA (LCACoA) 

were measured by mass spectrometry using internal standards for individual 

species. OGTT revealed signifi cant reductions in glucose tolerance in HF 

but not HF+N3 or NFD. Total ceramide was increased in HF (15.69±0.41 ng/ 

mg tissue) and HF+N3 (14.38±0.16) compared to NFD (13.05±0.43). Closer 

examination of individual ceramide species revealed that n3-PUFAs completely 

prevented HF-induced increases in medium chain saturated (C18) ceramides 

(NF: 7.38±0.31, HF: 9.41±0.27, HF+N3: 8.08±0.17 ng/mg tissue), but signifi cantly 

increased levels of long chain monounsaturated (C24:1) ceramides ceramides 

(NF: 2.74±0.06, HF: 2.45±0.06, HF+N3: 2.76±0.04 ng/mg tissue). Regression 

analyses revealed that C18 ceramide content is a strong predictor (R 2 =0.57, 

P


Obesity 

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1843-P 

Pyruvate Dehydrogenase Kinase 2 (PDK2) Defi ciency Reduces 

Fasting Blood Glucose Levels and Fat Accumulation in the Liver of 

Mice Fed a High Fat Diet 

YOUNGHOON GO, NAM HO JEOUNG, JIYUN JEONG, HYEON-JI KANG, KEUN- 

GYU PARK, EON-JU JEON, CHANG JOO OH, AE-KYUNG MIN, JEONG-KOOK KIM, 

SUN JOO LEE, HYUN-AE SEO, ROBERT A. HARRIS, IN-KYU LEE, Daegu, Republic 

of Korea, Indianapolis, IN 

Regulation of the activity of the pyruvate dehydrogenase complex 

(PDC) is critical for disposal of excess glucose, fuel selection by tissues, 

and conservation of substrates for glucose synthesis. A dominant role for 

PDK4 in down regulation of PDC activity is suggested by the remarkable up 

regulation of PDK4 during fasting. This hypothesis is supported by reduced 

fasting levels of blood glucose levels and three carbon gluconeogenic 

substrates in PDK4 knockout (PDK4 -/- ) mice. Relative to PDK4, less evidence 

has existed for an important role for PDK2 in glucose homeostasis. Although 

ubiquitously expressed, PDK2 is only modestly increased by fasting, and PDK2 

defi ciency has no effect upon fasting blood glucose levels in chow fed mice. 

However, in studies with mice fed a high fat diet for 16 weeks (12-13 mice 

per group), PDK2 defi ciency reduced body weight (WT vs PDK2 -/- , 49.8±0.8 vs 

42.8±1.2 g, Mean±SE, P

however their effects on energy balance were elucidated by our group recently 

showing that kinin B1r participates in the regulation of energy balance via a 

mechanism that could involve the modulation of leptin sensitivity. Now, using 

B2 knockout mice and in vivo techniques, we present here evidence of a novel 

role for kinin B2r in the regulation of mitochondrial activity and adiposity. 

Kinin B2r defi ciency in mice (B2-/-) resulted in less fat content, higher 

glucose uptake in isolated muscle (C), increased lean mass and robust 

protection against high fat diet (HFD)-induced weight gain (A). Under HFD 

B2-/- exhibited improved lipid oxidation and increased energy expenditure 

(B). Surprisingly, B2r defi ciency led to an augment in mitochondrial activity 

(E), fatty acid β-oxidation related gene expression (D) and superoxide 

dismutase activity in muscle (F). Finally, B2r mRNA levels were signifi cantly 

lower in 3T3-422A differentiated adipocytes (G) and during the early days of 

C2C12 myoblast differentiation (H). Together, our data suggest that kinin B2 

receptors participate in the regulation of energy balance via a mechanism 

that may involve the modulation of mitochondrial activity in muscle fi bers. 

Supported by: FAPESP, CNPq, CAPES 

1848-P 

Saturated and Unsaturated Fat-Induces Tissue-Specifi c Changes 

in Critical Nodes of Insulin Resistance in Diet-Induced Obese (DIO) 

Rats 

SURESH K. MOHANKUMAR, PETER ZAHRADKA, DANIELLE P. HANKE, LINDA 

SIEMENS, CARLA TAYLOR, Winnipeg, MB, Canada 

The liver and peripheral tissues including skeletal muscle and adipose 

play a pivotal role in the pathogenesis of insulin resistance (IR) induced by 

high fat diets (HFD). However, the originating nodes of IR in these tissues 

remain poorly understood. The present study therefore investigated the 

tissue-specifi c changes of protein markers of IR in response to isocaloric 

HFD of either saturated fat (SF) or unsaturated fat (USF) in obese-prone 

rats. Fasting serum analysis indicated that rats fed HFD for 12 weeks were 

insulin resistant with marked elevation in the fasting glucose (245±22 

mg/dL), insulin (45±5 µU/ml) and HOMA (27±5). Comparative analysis of 

gastrocnemius muscle (GM), liver (L) and epididymal adipose (EA) tissues 

indicated that the degree of relative phosphorylation/activation of the 

key nodes of IR was greater in some tissues versus others: IRS1-s 636/639 , 

GM>L=EA; IRS1-s 307 , GM>L=EA; AKT-s 473 , GM=EA>L; JNK, L>GM=EA; ERK, 

EA>GM=L, although there were no changes in the respective protein levels. 

These results suggest that activation of these critical nodes in response 

to high fat is tissue-specifi c. Next, we determined whether SF and USF 

differentially infl uence the critical nodes of IR. The HOMA value of SF and 

USF fed rats (37±4 and 19±2) indicated that SF fed rats were more insulin 

resistant than USF. However, there were no differences between SF and USF 

in the activation of the above-mentioned IR nodes. Interestingly, the animals 

fed USF had active tyrosine phosphorylation of AKT-t 308 , a positive regulator 

of insulin signaling, in GM and EA (64% and 32% increase versus SF) despite 

no differences in caloric intake. These results paralleled the reductions of 

fasting glucose, insulin and HOMA in USF fed rats (-27, -31 and -48% versus 

SF group), suggesting that USF sustains metabolic functions of GM and EA 

tissues. In summary, these data establish IRS1/AKT in muscle, JNK in liver 

and AKT/ERK in adipose tissue as key players in the pathogenesis of IR in 

response to HFD and USF delays the progression of IR via AKT. 

Supported by: Canola Council of Canada 




CATEGORY 

A499 

1849-P 

The Contribution of Enhanced Myocardial mTOR/S6K1 Activation 

to Impaired Insulin Signaling, Oxidative Stress and Diastolic 

Dysfunction in Obese db/db Mice 

VINCENT G. DEMARCO, JAVAD HABIBI, LAKSHMI PULAKAT, ADAM T. WHALEY- 

CONNELL, LIXIN MA, MELVIN R. HAYDEN, ROGER D. TILMON, C.B. KRUEGER, NA- 

THAN REHMER, MONA GARRO, MING YANG, JAMES R. SOWERS, Columbia, MO 

Obesity, the metabolic syndrome and heart failure (HF) are epidemics in 

the United States. Obesity related HF is characterized by decreased insulin 

metabolic signaling and diastolic dysfunction. We have investigated the role 

of mTOR/S6K1 activation in impairment of insulin metabolic signaling and 

diastolic relaxation in young db/db mice. The initial diastolic fi lling rate, as 

measured by high resolution MRI, was decreased in 12 week hyperglycemic 

db/db mice compared to age matched controls. There were contemporaneous 

increases in reactive oxygen species (ROS), increased small abnormal 

mitochondria, lipid accumulation and interstitial fi brosis in db/db heart. There 

was increased Thr 389 phosphorylation (P)/activation of S6K1, enhanced 

association of S6K1 with IRS-1 and Ser307 P of IRS-1. This was associated 

with decreased total IRS-1 and a tendency (not signifi cantly) for decreased 

P/activation of Akt. This is consistent with fi ndings in other tissues where 

S6K1 P has been associated with Ser P and associated ubiquinization/ 

degradation of IRS-1. A mitochondrial source of increased ROS is likely as 

the number of mitochondria was increased and NADPH oxidase activity was 

not increased in db/db heart. In summary, young db/db mice manifested 

diastolic dysfunction possibly due to decreased insulin metabolic signaling, 

increased mitochondrial generation of ROS, and interstitial fi brosis. 

Supported by: NIH R01-HL-63904 and VA Merit (JRS), and VA Career Development 

Award (AWC) 

1850-P 

The Inhibition of Cannabionoid Receptor CB1 Increases GLUT4 

Expression in Adipocytes and Adipose Tissue 

DANIELA T. FURUYA, ANA C. POLETTO, HELAYNE S. FREITAS, UBIRATAN F. 

MACHADO, São Paulo, Brazil 

Evidences have suggested that the endocannabinoid system is overactive 

in obesity, resulting in enhanced endocannabinoid levels in both circulation 

and visceral adipose tissue. The cannabinoid CB1 receptor is expressed 

in the adipose tissue besides the brain. Few studies in vitro suggest that 

CB1 activation increases glucose uptake in adipocytes. The objective of the 

present study was to investigate the CB1 receptor modulation on glucose 

transporter GLUT4 expression and the related mechanisms in adipocytes and 

adipose tissue of obese mice. For this, 3T3-L1 adipocytes were incubated 

in the presence of a selective agonist of CB1 receptor (1 mM ACEA) or a 

selective antagonist of CB1 receptor (0.1, 0.5 or 1 mM AM251) or both. After 

2, 4 or 24 hours, cells were harvest to evaluate GLUT4 mRNA (Real Time 

PCR) and protein (Western blotting), and NFkappaB activation specifi cally 

in the promoter of SLC2A4 gene (EMSA) which encodes GLUT4 protein. 

Additionally, insulin sensitivity in vivo and GLUT4 protein from adipose tissue 

were assessed in monosodium glutamate–induced obese mice untreated or 

treated with AM251 (0.01 mg/kg body weight). Acute and chronic incubation 

with AM 251greatly increased GLUT4 protein content [0.1, 0.5 or 1 µM AM251 

for 4 hours treatment, 309%, 313% or 357% vs C, respectively, P


Obesity 

POSTERS 

body fat content in dietary-induced obese rats (DIO rats). PPARg agonists, 

like Pioglitazone (Pio), are effi cacious drugs for the treatment of type 2 

diabetes. However, a common side effect of this drug class is weight gain. 

Here, we investigated whether BI 10773 could attenuate Pio-induced 

weight gain of DIO rats after 4 weeks treatment with 10mg/kg BI 10773, 

10mg/kg Pio and the combination of both (n=10 per group). Pio signifi cantly 

increased body weight (+6.8 %), whereas BI 10773 led to weight loss (-3.1%) 

compared to control animals. Animals that received BI 10773 in combination 

with Pio had no signifi cant increase in body weight (+1.6% vs. vehicle). 

As DIO rats are insulin-resistant and hyperinsulinemic we investigated 

the effect of the combination on metabolic parameters. Both compounds 

signifi cantly reduced fed plasma insulin levels (vehicle: 5.27 ± 0.7; BI 10773: 

3.53 ± 0.4; Pio: 2.27 ± 0.3 ng/ml) which was further reduced (p=0.054) by the 

combination (BI 10773 + Pio: 1.55 ± 0.1 ng/ml). Furthermore, the combination 

of BI 10773 + Pio reduced plasma triglyceride levels, whereas either 

compound alone had no effect. As expected, BI 10773 and BI 10773 + Pio 

markedly increased urinary glucose excretion. 

After termination of the study carcass analysis was performed to assess 

body composition. The fat content was 147.4 + 7.8 g in the vehicle group 

compared to 140.7 + 7.1 g (BI 10773). Pio increased body fat content to 

170 + 6.8 g but the gain in body fat was prevented by combination treatment 

(BI 10773 + Pio: 149.6 + 7.2 g). 

Our data show that BI 10773 prevented the Pio-induced body weight gain 

and the associated gain of body fat after 4 week treatment of DIO rats. 

Furthermore, BI 10773 improved hyperinsulinemia of DIO rats alone and in 

combination with Pio. Our data support the concept of a combination of BI 

10773 with Pio for the treatment of type 2 diabetes. 

1852-P 

Tissue-Specifi c Dysregulation of Hexose-6-Phosphate Dehydrogenase 

and Glucose-6-Phosphate Transporter Expression in a Mouse 

Model of Type 2 Diabetes 

YANJUN LIU, YUICHI NAKAGAWA, YING WANG, WEI WANG, JUAN ORTEGA, 

THEODORE C. FRIEDMAN, Los Angeles, CA, Hamamatsu, Japan 

Tissue-specifi c amplifi cation of glucocorticoid action through 11β-hydroxysteroid 

dehydrogenase type 1 (11β-HSD1) affects the develop ment of the 

metabolic syndrome. 

Hexose-6-phosphate dehydrogenase (H6PDH) mediates intracellular 

NADPH availability for 11β-HSD1 and depends on the glucose-6-phosphate 

transporter (G6PT). To assess whether the tissue-specifi c alterations of 

H6PDH and G6PT expression could contribute to local glucocorticoid action 

and insulin resistance in type 2 diabetes and obesity, we characterized the 

role of H6PDH and G6PT in pre-receptor metabolism of glucocorticoids by 

examining the production of the hepatic 11β-HSD1-H6PDH-G6PT system in 

a mouse model of type 2. 

We observed that increased production of hepatic H6PDH in diabetic 

db/db mice was paralleled by upregulation of hepatic G6PT production and 

elevated circulating levels of corticosterone. In contrast, pharmacological 

inhibition of H6PDH and G6PT expression decreased NADPH production 

accompanied by reduction of 11β-HSD1 in the liver and subcutaneous fat, 

and attenuated the phenotype of type 2 diabetes. Incubation of mouse 

hepatocytes with corticosterone enhanced G6PT and H6PDH production 

with corresponding activation of 11β-HSD1 and PEPCK. Knockdown of 

H6PDH by siRNA attenuated the corticosterone-induced H6PDH production 

in these intact cells. Addition of the G6PT inhibitor to primary hepatocytes 

suppressed H6PDH production. These fi ndings suggest that increased 

hepatic and adipose H6PDH and G6PT expression may contribute to 

11β-HSD1 upregulation of local glucocorticoid action that may be related to 

the development of type 2 diabetes. 

Supported by: NIDDK SC1DK087655 

1853-P 

TrkB Agonist Induces Body Weight Loss by Activating Satiety Centers 

in the Brain 

TIFFANY GARESKI, SARAH WILL, DAVID KUBASIAK, THADDEUS UNGER, 

KIMBERLY COUGHLAN, GUO FENG, YING SUN, XIANGPING LI, ARIFUL QADRI, 

DARRELL PANZA, SEUNG HAHM, JULI JONES, JANET PAULSEN, RUTH GIMENO, 

MYLENE PERREAULT, Cambridge, MA 

Brain-derived neurotrophin factor (BDNF) and its receptor, tropomyosinrelated 

kinase B (TrkB), have emerged as critical regulators of central neural 

circuits involved in energy homeostasis. 

The potential therapeutic application of TrkB activation has been 

demonstrated in several rodent models in which BDNF administration 

induced weight loss mainly through appetite suppression. In this study, we 


OBESITY—HUMAN 

CATEGORY 

A500 

sought to determine the biological effects of a TrkB agonist and identify its 

mechanism of action. 

Diet-induced obese (DIO) mice were injected with a potent and highly 

selective mouse monoclonal TrkB agonist and the effects on body weight 

and food intake were determined. We observed that TrkB activation induced 

robust and reversible weight loss accompanied by a concomitant reduction in 

food intake. To gain further insight into the hypophagic effect of the agonist, 

we studied circadian meal patterns and observed that TrkB agonism reduced 

the amount of food ingested per meal but did not affect meal frequency. 

In addition, pair-feeding experiments showed that body weight was mostly 

regulated by food intake. Given the high selectivity of the blood-brain-barrier, 

it is unknown whether the effects of this TrkB agonist on food intake are 

mediated through activation of TrkB receptors in the central nervous system 

(CNS) or periphery. To address this, we peripherally injected fl uorescently 

labeled TrkB agonist and detected fl uorescence in the medium eminence and 

the arcuate nucleus of the hypothalamus as well as the dorsal vagal complex 

of the hindbrain. Collectively, our studies show that TrkB agonism decreases 

body weight mainly by increasing satiety through direct activation of the 

brain satiety centers. 

1854-P 

Weight Control Diminishes Progression of Insulin Resistance Due 

to Estrogen-Deprived Condition 

MUJALIN PRASANNARONG, KANOKWAN VICHAIWONG, VITOON SAENGSIRI- 

SUWAN, Bangkok, Thailand 

Prolonged estrogen deprivation in ovariectomized rats resulted in the 

phenotypic features of the metabolic syndrome including increased visceral 

fat accumulation, dyslipidemia, impaired glucose tolerance, and insulin 

resistance of skeletal muscle. Because these animals are also hyperphagic, 

the development of metabolic defects under estrogen-deprived condition 

could be confounded by excessive calorie consumption. To determine the role 

of calorie consumption in the development of the phenotypic characteristics 

of the metabolic syndrome in animal under prolonged estrogen deprivation, 

adult female Sprague-Dawley rats were randomly assigned to shamoperated 

(SHAM), ovariectomized (OVX), pair-fed ovariectomized (OVX+PF), 

or body weight-controlled ovariectomized (OVX+WC) groups. After a 12week 

experimental period, whole-body glucose tolerance, skeletal muscle 

insulin action, intra-abdominal fat content and serum lipid profi le were 

evaluated. When compared to OVX group, both pair-feeding and weight 

control paradigms signifi cantly reduced intra-abdominal fat accumulation by 

32% and 44% (p

esistance and type 2 diabetes. By comprehensive proteomic profi ling of 

the human adipocyte secretome we identifi ed DPP4 as a novel adipokine. 

This study assessed the association of the adipokine DPP4 to the metabolic 

syndrome. 

Fully differentiated adipocytes exhibit a substantially higher release of 

DPP4, when compared to preadipocytes or macrophages. DPP4 release 

from adipocytes is increased after treatment with insulin and TNFα. Direct 

addition of DPP4 to adipocytes impairs insulin signaling at the level of 

insulin-stimulated Akt phoshorylation. A signifi cantly higher level of DPP4 

protein expression was seen in visceral as compared to subcutaneous fat of 

obese patients. DPP4 expression in lean subjects was signifi cantly lower in 

both depots compared to the obese patients and with no regional difference. 

DPP4 serum concentrations signifi cantly correlated with adipocyte size, 

serum leptin and various clinical parameters related to the metabolic 

syndrome as BMI, fasting insulin and hs-CRP. Using adipose tissue explants 

from lean and obese subjects, we observed a 2-fold increase in DPP4 release 

in obesity that strongly correlated with adipocyte volume and parameters of 

the metabolic syndrome. DPP4 release from adipose tissue was decreased 

to the lean level after weight reduction of the obese patients induced by 

bariatric surgery. Both circulating DPP4 and DPP4 released from adipose 

tissue strongly correlated positively with an increasing risk score for the 

metabolic syndrome. 

DPP4 is a novel adipokine which may be linked to adipocyte hypertrophy 

and adipose tissue infl ammation. As DPP4 release strongly correlates with 

adipocyte size, adipose tissue could represent an important source of DPP4 

in obesity. We therefore suggest that DPP4 may be critical in linking adipose 

tissue and the metabolic syndrome. 

& 1856-P 

Lipid Accumulation Products Index as a Better Predictor for Insulin 

Resistance and Glucose Intolerance Than BMI in Young Obese Women 

with Polycystic Ovary Syndrome 

JEE-YOUNG OH, HYE JIN LEE, YOUNG SUN HONG, YEON-AH SUNG, Seoul, 

Republic of Korea 

Elevated waist circumference and elevated triglycerides has been 

considered as a surrogate marker of cardiovascular risk. BMI may not to 

be the best marker for obesity-related disease. Lipid accumulation product 

(LAP) index, an ordinal scale combining waist circumference and triglycerides 

concentration, was suggested as a better marker for cardiovascular disease 

than BMI. 

We examined whether the LAP index is associated with cardiovascular 

risk factors and a better predictive marker than BMI in obese women with 

polycystic ovary syndrome (PCOS). We recruited 223 obese (BMI ³23kg/m 2 ) 

women with PCOS and 230 age-, and BMI-matched regular cycling control 

women by voluntary participation for the prevalence study. PCOS was 

diagnosed by using NICHD, and LAP index was calculated using the formula 

[waist circumference(cm)-58] x TG(mmol/L). Glucose intolerance was defi ned 

as IFG, IGT or diabetes, and insulin resistance as HOMA value greater than 

90 percentile of obese controls. 

Mean age and BMI were 24±4 yr and 25.6±2.2 kg/m 2 in PCOS and 24±4 yr 

and 25.2±1.8 kg/m 2 in controls. LAP index was signifi cantly higher in women 

with PCOS than controls (32.2±21.9 vs. 23.6±15.6, P


Obesity 

POSTERS 

& 1859-P 

Liraglutide Provides Weight Maintenance and Additional Weight 

Loss after Low Calorie Diet-Induced Weight Loss in Obese Subjects 

without Diabetes: The SCALE Maintenance Study 

THOMAS A. WADDEN, PRISCILLA HOLLANDER, SAMUEL KLEIN, KEVIN NIS- 

WENDER, VINCENT WOO, PAULA HALE, TU DUYEN LE THI, LOUIS J. ARONNE, 

Philadelphia, PA, Dallas, TX, St. Louis, MO, Nashville, TN, Winnipeg, MB, Canada, 

Princeton, NJ, Copenhagen, Denmark, New York, NY 

Weight loss is diffi cult to achieve and sustain by lifestyle therapy alone. 

Liraglutide, a once-daily human glucagon-like peptide-1 analog approved 

for the treatment of T2DM, induced dose-dependent wt loss in obese 

subjects without diabetes in a phase 2 study, with highest effi cacy shown 

by a 3mg/d dose. The present randomized, placebo-controlled trial tested 

the ability of liraglutide (3mg/d) to maintain diet-induced wt loss. Subjects 

who lost >5% body wt after a 4–12 week run-in (RI) with a low-calorie diet 

(1200–1400kcal/d) and exercise counseling were randomized to receive 

liraglutide or placebo plus a 500kcal/d defi cit diet and exercise regimen for 

56 weeks. Of 511 subjects entering RI, 422 were randomized 1:1 to the 2 

arms. The mean RI wt loss for all randomized subjects was 6.0% (6.3kg). 

Over the next 56 weeks, additional wt loss occurred post-randomization (PR) 

(6.1% vs 0.05%, liraglutide vs placebo, p

with ≥1 post-baseline weight on study drug). More NB32-treated women (vs. 

placebo) achieved weight loss of ≥5% (46.5 vs. 23.8%; p


Obesity 

POSTERS 

the confounding effect of differential use of rescue medications, the last 

available glycemic measure prior to initiation of rescue medication was 

carried forward in the following post hoc analyses for glycemic parameters: 

Compared to PBO, NB32 improved HbA1c (-0.6% vs 0.1%; p

& 1870-P 

Relationship between Total Body Fat and Body Fat Distribution and 

Left Ventricular Diastolic Dysfunction in Men with the Metabolic 

Syndrome 

ANNIE MORIN, JEAN-PIERRE DESPRÉS, MARIE ARSENAULT, MARIE-KRISTELLE 

ROSS, JEAN BERGERON, ANGELO TREMBLAY, NATHALIE ALMERAS, PAUL POIRIER, 

Quebec, QC, Canada 

Left ventricular diastolic dysfunction (LVDD) has been observed in 

subjects with metabolic syndrome. We examined the relationship between 

several indices of adiposity (waist circumference, waist-to-hip ratio, waistto-height 

ratio, body mass index, body fat estimated by bioimpedance, and 

visceral adiposity assessed with computed tomography) and LVDD in men 

with metabolic syndrome without heart disease. A total of 76 men between 

30-65 years (age 48.5 yrs) evaluated by echocardiography for the presence 

of LVDD, using doppler mitral infl ow with/without Valsalva maneuver, 

pulmonary venous fl ow and tissue doppler imaging, were included. LVDD 

was graded according to guidelines. All had normal left ventricular ejection 

fraction. Twenty-one subjects were classifi ed as normal, whereas 55 were 

classifi ed as abnormal in terms of LVDD, including 17 grade I, 33 grade 2, 

and 5 as unclassifi ed. Groups were comparable regarding adiposity indices. 

Visceral adipose tissue volume was also similar between groups: normal 

diastolic function (mean volume 1778±480 cm 3 ) vs. LVDD (1938±460 cm 3 ); 

grade I (1924±531 cm 3 ) vs. grade II (1923±439 cm 3 ). However, subcutaneous 

adipose tissue volume was different between groups; 2121±828 cm 3 for the 

group with normal function vs. 1698±499 cm 3 for LVDD (p=0.04). There was 

no signifi cant difference between grades I and II. Ratio of visceral adipose 

tissue volume on total adipose tissue volume was also statistically different 

between normal function (0.47±0.11) and LVDD (0.53±0.07) (p=0.01). Again, 

no signifi cant difference was found between grades I and II. Thus, after 

adjustment for age, no adiposity indices explained the presence or the 

severity of LVDD. Furthermore, there was no difference regarding fasting 

glucose, insulinemia, C-peptide, insulin resistance, lipid profi le, C-reactive 

protein and adiponectin between groups. In conclusion, with age taken 

into consideration, we did not observed any signifi cant relationship 

between indices of adipose tissue distribution and LVDD in our sample of 

asymptomatic men with features of the metabolic syndrome. 

Guided Audio Tour: Pharmacologic Treatment of Obesity (Posters 1871-P 

to 1878-P), see page 15. 

& 1871-P 

Reduction in the Expression of β-Amyloid Precursor Protein (APP) 

Following Roux-en-Y Gastric Bypass Surgery (RYGB) and Weight Loss 

PARESH DANDONA, HUSAM GHANIM, SCOTT MONTE, CHANG LING SIA, 

JEROME SCHENTAG, SANDEEP DHINDSA, JOSEPH CARUANA, Buffalo, NY 

Obesity and type 2 diabetes are associated with an increase in the 

incidence and prevalence of Alzheimer’s disease (AD). We have recently 

demonstrated that peripheral blood mononuclear cells (MNC) express APP, 

the precursor of β-amyloid, which forms the pathognomonic plaques in the 

brain. We hypothesized that APP expression diminishes after the marked 

caloric restriction and weight loss associated with RYGB. Fifteen type 2 

diabetic patients with morbid obesity underwent RYGB following which 

their caloric intake diminished and they lost 38.5 ±2.9 Kg in weight over 6 

months. BMI fell from 54.4±3.1 to 40.5± 2.9 Kg/m 2 . There was a signifi cant 

fall in plasma concentrations of glucose (from 148±8 to 101±4mg/dl), insulin 

(from 18.5±2.2 to 8.6±1.0µU/ml) and HOMA-IR (from 7.1±1.1 to 2.1±0.3).). The 

expression of APP mRNA fell by 31±9% and that of protein fell by 22±12%. In 

addition, there was a reduction in other AD related genes including presinilin 

2 (PN2) which fell by 27±10%, ADAM-9 which fell by 35±12% and GSK-3β 

which fell by 28±6% (P


Obesity 

POSTERS 

before 63 ± 8 vs. after 69 ± 8 bpm, p = 0.049). Low frequency gain (LF-gain), 

a measure for barorefl ex sensitivity, decreased after the HD (LF-gain before 

13.0 ± 9.7 vs. after 9.5 ± 6.1 msec/mmHg, p = 0.017). HF variability (HFvar), 

primarily determined by respiratory vagal functions, decreased after the HD 

(HFvar before 496 ± 795 vs. after 354 ± 519 msec², p = 0,050). LF/HF ratio, an 

indicator of sympathovagal balance, did not change after the HD. 

Our results indicate that a hypercaloric diet induces insulin resistance in 

lean healthy men and has signifi cant effects on cardiovascular autonomic 

indices which point to changes towards sympathetic predominance/vagal 

withdrawal. This suggests that functional changes in the autonomic nervous 

system are part of the adaptive response to a hypercaloric milieu and may in 

part mediate the changes in glucose metabolism. 

& 1874-P 

Naltrexone SR/Bupropion SR Combination Therapy Improves Control 

of Eating and Reduces Food Cravings in Overweight and Obese Subjects 

JAMES O. HILL, HOLLY WYATT, SONJA K. BILLES, COLLEEN BURNS, RAUL 

HARRIS-COLLAZO, EDUARDO DUNAYEVICH, JOHN BLUNDELL, Denver, CO, La 

Jolla, CA, Leeds, United Kingdom 

The effects of naltrexone/bupropion combination therapy (NB) on obesity 

and eating behavior (Control of Eating Questionnaire: CoEQ, twenty 100mm 

visual analog scales) were evaluated in obese/overweight individuals in 

the COR-II study at baseline and Weeks 8, 16, 28, and 56. This Phase 3, 

double-blind, placebo-controlled, 56-week study, randomized 1496 subjects 

in a 2:1 ratio to NB32 (32mg naltrexone sustained-release [SR]/360mg 

bupropion SR) or placebo. The completion rate was 54% in each group. 

Baseline characteristics for the modifi ed ITT-LOCF population (subjects with 

≥1 post-baseline weight on study drug) were: 84% female, 85% Caucasian, 

mean±SD age 44±11y, weight 100±16kg, and BMI 36±4kg/m 2 . Week 56 

weight loss was greater (p

placebo-controlled 56-week study. Subjects (N=1496) were randomized 2:1 to 

NB32 (32mg naltrexone SR/360mg bupropion SR) or placebo. The completion 

rate was 54% in each group. Baseline characteristics for the modifi ed ITT- 

LOCF population (subjects with ≥1 post-baseline weight on study drug) were: 

84% female, 85% Caucasian, mean±SD age 44±11y, weight 100±16kg, BMI 

36±4kg/m 2 , IWQOL-Lite Total score 73±17 (moderate impairment), IWQOL- 

Lite subscales: Physical Function 70.4±20.4, Self-Esteem 56.2±25.6, Sexual 

Life 75.5±26.3, Public Distress 86.6±18.0, Work 87.1±17.1, and SF-36: 

Physical Component 50.2±7.1 and Mental Component 54.4±7.3 (a score of 50 

is equivalent to average health). Week 56 weight loss was greater (p


Obesity 

POSTERS 

rs29941 (KCTD15; p=0.006) and rs7561317 (TMEM18; p=0.043) had effect 

also on waist-hip ratio. The carriers of minor alleles G in homozygote forms 

in variants rs4923461 (BDNF; p=0.036) and rs1800592 (UCP1; p=0.033) had 

increased muscle mass in body composition, furthermore, GG homozygotes 

in rs1800592 had decreased fat mass (p=0.037) and also increased musclefat 

ratio (p=0.026). In summary, we confi rmed associations of majority of the 

10 obesity-susceptibility variants with several obesity-related parameters; 

the most complex infl uence on anthropometric features was identifi ed in 

variant rs7561317 in TMEM18 gene. 

Supported by: IGA MHCR NS/9839-4 and NS10209-3/2009 

1882-P 

Associations of Waist Circumference and Body Mass Index with 

Fasting Blood Glucose among Overweight African American Parishioners 

in a Southern Semi-Urban Community 

RICHARD W. SATTIN, LOVORIA B. WILLIAMS, JAMES K. DIAS, THOMAS JOSHUA, 

LUCY N. MARION, Augusta, GA 

African-Americans (AAs) are at high risk for type 2 diabetes (T2D) partly 

because rates of obesity are about twice as high as that in the overall US 

population. Although body mass index (BMI) has been considered an accurate 

clinical measure of body fat, central adiposity, approximated by waist 

circumference (WC), has been found to be a reliable predictor of metabolic 

disturbances, morbidity and mortality. Our objective is to determine, in a 

cohort of overweight AAs participating in the Fit Body and Soul Study (FB&S), 

the statistical relationships between WC, BMI, demographic characteristics, 

and fasting blood glucose (FBG). FB&S is a single-blinded, cluster randomized, 

community trial developed to test the effectiveness of a faith-based 

adaptation to the Diabetes Prevention Program. AA church parishioners, in 

a semi-urban Southern US community aged 20 to 64 years, with a BMI of 

25 kg/m 2 or greater and without a self-reported history of diabetes, were 

eligible to participate. Baseline demographic and measurement data from 

the participants in the fi rst ten of 20 churches available were collected, 

including age, gender, BMI, WC, and FBG. Standard descriptive statistics 

were calculated and multiple linear regression and correlation analyses 

were conducted to model FBG. From September 2009 through August 2010, 

393 AAs were consented. Eighty-four percent were female; the average 

age of those consented was 46.8 years (95% confi dence interval (CI): [45.6, 

47.8]); the average BMI was 35.4 kg/m 2 (95% CI: [34.7, 36.2]); the average 

WC was 107.7 cm (95% CI: [106.1, 109.4]); the average FBG was 93.0 mg/ 

dl (95% CI: [91.4, 94.5]). The partial correlation of FBG with BMI, adjusted 

for age and gender, was 0.189 (p-value < 0.001). The partial correlation of 

FBG with WC, adjusted for age and gender, was 0.192 (p-value < 0.001). Our 

fi ndings suggest that both BMI and WC are important contextual variables 

and provide similar results for the study of T2D in AAs. 

Supported by: NIH grant DK082401 

1883-P 

Bariatric Surgery in a Multi-Ethnic WITHDRAWN 

Type 2 Diabetes Population 

BRANDON J. ORR-WALKER, MARIE WHITE, IRENE ZENG, Auckland, New Zealand 

This study is to determine the effect of adjunctive support compared with 

standard care in a morbidly obese, diabetic cohort on body mass index (BMI) 

undergoing publicly-funded bariatric surgery with secondary endpoints of 

glycemic control (HbA1c), blood pressure, lipids, pulmonary function and 

quality of life. As the study has not yet been completed, unblinded results 

of recruitment and clinical response for completers to 1 year post surgery 

are presented. All subjects have type 2 diabetes on hypoglycaemic agents 

and are enrolled in a diabetes chronic care management program (CCM) 

which provides free quarterly review in primary care, in addition to the usual 

funded laboratory, pharmaceutical and hospital services. 

Entry to the study was for those enrolled for at least one year in CCM 

with a BMI ≥35, 7.0≤ HBA1c ≤10.0 on hypoglycaemic medication, serum 

creatinine ≤150 (≤ 120 for women) and urinary albumin creatinine ratio ≤ 

200 mg/mmol. Additional surgical criteria limiting surgery to non-smokers 

aged 20-60, with a weight of ≤160kg were applied resulting in a potential 

recruitment population of 1924 patients from the total 7367 enrolled. (27% 

Maori, 55% Pacifi c, 11% European, 7% Asian). Informed consent for both 

surgery and the study was required for inclusion. 

Of those referred to the study team by their family practitioner, 113 were 

not enrolled( 63 did not meet the study criteria, 9 had other contraindications, 

34 declined surgery, 2 declined study involvement but had surgery, others 5), 

and 9 withdrew after randomisation. Overall Pacifi c and Asian peoples are 

underrepresented by referral, resulting in lower inclusion in the study. Rates 

of non-enrolment were similar. Enrolees had a mean age of 48 years, and 

64% were female, 33% Maori, 30% Pacifi c, 35% European and 1% Asian. Of 



CATEGORY 

A508 

those completing the study so far(n=34) BMI fell from 49 ± 7 to 34 ± 4kg/m 2 , 

and weight from 137 ± 22 to 96 ± 15 kg. Median HBA1c fell from 7.8 (IQR 7.2, 

9.1) to 5.9 (IQR 5.4, 6.4). 

Bariatric surgery provides early clinical benefi ts to a patient group already 

participating in fully-funded proactive chronic disease management for 

diabetes. Referral for surgery appears to be lower for Pacifi c and Asian 

peoples. 

1884-P 

Basal Endogenous Glucose Production and Insulin Levels Are 

Reduced 2 Weeks after Bariatric Surgery with No Effect on Hepatic 

and Peripheral Insulin Sensitivity 

BARBARA A. DE WEIJER, EDO AARTS, IGNACE M. JANSSEN, ARNOLD VAN DE 

LAAR, KARIN KAASJAGER, MARIETTE T. ACKERMANS, ERIC FLIERS, MIREILLE 

J. SERLIE, Amsterdam, The Netherlands, Arnhem, The Netherlands 

Bariatric surgery has very early metabolic effects on glucose metabolism 

before the occurrence of clinically signifi cant weight loss. This suggests an 

acute effect of the surgery itself, i.e. bypassing the nutrient fl ow from the 

proximal gastrointestinal tract. We hypothesized that Roux-en Y gastric 

bypass surgery (RYGB) ameliorates glucose metabolism by increasing 

hepatic and peripheral insulin sensitivity. 

We studied glucose metabolism and lipolysis in the basal state and during 

a hyperinsulinemic euglycemic clamp using stable isotopes two weeks 

before and two weeks after RYGB. 

We included 12 pre-menopausal Caucasian women (median age 38 [26- 

49] yrs, median BMI 45 [39.2-53.9] kg/m2) scheduled for RYGB. Two weeks 

after RYGB median weight loss was 5.5kg [2-14 kg]. 

Basal insulin and glucose levels decreased after surgery (insulin presurgery 

73 [21-122] vs post-surgery 48 [15-120] pmol/L, p = 0.0025 and 

glucose pre-surgery 5.5 [4.4 – 6.9] vs post-surgery 4.8 [3.9 – 5.7] mmol/L, 

p = 0.0058 resp.). Endogenous glucose production (EGP) was lower after 

surgery (before 13.5 [11.3- 15.9] vs after 11.4 [10.3 – 14.2] μmol/kgFFM/ 

min, p = 0.0078). Insulin levels were lower during the clamp after surgery, 

suggesting enhanced clearance. Hepatic and peripheral insulin sensitivity 

(both corrected for insulin) did not change after surgery. FFA were increased 

after surgery in the basal state (0.76 [0.67–1.13] vs 0.94 [0.73-1.33] mmol/L, 

p = 0.0049) and during the fi rst step of the clamp (0.13 [0.02 – 0.29] vs 0.34 

[0.06 – 0.49] mmol/L, p = 0.0010). Lipolysis expressed per REE tended to 

increase in the basal state (259 [182 - 426] vs 257 [167 - 384] µmol/kcal, p = 

0.0674), and was higher during hyperinsulinemia (91 [70 - 298] vs 132 [100 - 

403] µmol/kcal, p = 0.0020). 

Within 2 weeks, bariatric surgery reduces basal EGP, insulin and glucose 

levels but has no acute benefi cial effect on hepatic or peripheral insulin 

sensitivity. The latter may be explained by higher rates of lipolysis and 

exposure to FFA induced by the hypocaloric state. 

1885-P 

CREW (Calcium REduces Weight) Study: Impact of Calcium Supplementation 

on Weight, Adiposity, Glycemia and Hypertension 

LAKSHMI KANT SHANKHDHAR, KSHITIJ SHANKHDHAR, UMA SHANKHDHAR, 

SMITA SHANKHDHAR, Lucknow, India 

Aims: CREW study aimed to observe impact of Calcium supplementation 

(CaS) on wt, waist circumference (WC), body fat% (BFP), BP, glycemia in T2 

diabetic (2D) & nondiabetic (ND) women. 

Methods: 120 drug naïve hypertensive obese female patients, both ND 

& 2D, were divided into 2 control (NDCo&2DCo) & 2CaS receiving (NDCa 

and 2DCa) subgroups, with age (ND=44.6±9.86, 2D=50.85±7.37yrs), Wt 

(ND=94.7±9.07, 2D=89.0±8.07Kg), BMI (ND=37.91±4.41, 2D=36.75±3.37Kg/ 

M2), WC (ND=102±6.44, 2D=100.9±4.08Cm) & BFP (ND=41.0±5.26, 2D= 

39.96±3.0%). They were advised 300 lesser calories, 30min brisk walk for 3 

months along with education. 

Results: NDshowed more reduction than 2D in Wt (NDCo=3.6 vs 

2DCo=1.13%, NDCa=5.78 vs 2DCa=5.23%), WC (NDCo=1.91 vs 2DCo=0.59%, 

NDCa=1.76 vs 2DCa= 1.58%) & BFP (NDCo=4.87 vs 2DCo=1.63%, NDCa=5.36 

vs 2DCa=3.33%). CaS reduced SBP more (2DCa=8.2 Vs 2DCo=3.03%) than 

DBP (2DCa=2.92 vs 2DCo=1.26%) in diabetics. Glycemia improved more with 

CaS:2DCa (FBG=19.08, PPBG=25.9, A1c=8.8%) vs 2DCo (FBG=7.5, PPBG=8.8, 

A1c= 4.9%) 

Conclusions: CaS plays positive role in reduction of adiposity, glycemia 

& BP. 

& Guided Audio Tour poster ADA-Funded Research

Observations: 

NDCo (n=30) NDCa (n=30) 

Baseline Mean Final Mean p value Baseline Mean Final Mean p value 

Wt (Kg) 96.10±7.76 92.70±8.42 0.004 93.30±10.45 87.90±11.96 0.0003 

BMI (Kg/M2 ) 38.31±4.28 36.74±4.62 0.006 37.51±4.28 35.28±4.06 0.0004 

WC (Cm) 99.00±4.00 97.10±5.13 0.003 102±6.61 100.50±7.26 0.0014 

BFP (%) 41.01±4.70 38.98±4.90 0.002 40.99±6.02 38.64±5.42 0.00003 

SBP (Hg mm) 132.80±5.67 127.60±4.19 0.003 131.00±7.37 126±.0.00 0.0263 

DBP (Hg mm) 80.60±1.34 78.80±2.14 0.0148 81.00±3.16 78.80±3.15 0.0646 

2DCo (n=30) 2DCa (n=30) 

Wt (Kg) 88.30±8.52 87.30±8.80 0.0236 89.70±8.00 85.00±8.11 0.000003 

BMI (Kg/M 2 ) 36.93±3.19 36.49±3.07 0.0250 36.58±3.70 34.65±3.49 0.000003 

WC (Cm) 101.00±2.86 100.40±2.45 0.0025 100.80±5.20 99.20±5.15 0.00002 

BFP (%) 40.32±2.16 39.66±2.08 0.0002 39.61±3.74 38.11±3.40 0.00000 

SBP (Hg mm) 132.40±6.31 128.20±3.45 0.0358 133.80±10.47 122.40±4.50 0.0014 

DBP (Hg mm) 78.80±3.15 77.80±4.15 0.1231 82.00±4.32 79.60±0.84 0.052 

FBG (mg%) 121.20±20.24 112.00±8.76 0.0588 131.00±18.01 106.00±7.45 0.0028 

PPBG (mg%) 153.60±21.64 139.00±16.94 0.0276 165.60±27.93 122.70±10.01 0.0006 

A1c (%) 7.5±0.22 7.1±0.18 0.0002 7.67±0.43 6.99±0.20 0.00006 

1886-P 

Depot-Specifi c Expression of Adipogenic Genes in Subcutaneous 

and Omental Adipose Tissues of Women and Their Relationship with 

Dyslipidemia 

HONGYUN LU, XIAOFENG LI, PANWEI MU, JIANG WEI, LONGYI ZENG, Guangzhou, 

China 

Depot differences in adipose tissue function may have implications 

for insulin resistance (IR), the adipogenesis inability of the subcutaneous 

adipose tissue (SAT) may cause ectopic fat deposition and IR, but the 

molecular mechanism remains unclear. This study aimed to measure gene 

expression involved in adipocytes differentiation and adipokines secretion 

in human SAT and omental adipose tissue (OAT), and then investigate their 

relationship with obesity and IR. We examined the expression of candidate 

genes from 29 lean and 12 overweight non-menopausal women. Serum 

leptin, adiponectin, retinol-binding protein 4(RBP4) and fasting insulin (FINS) 

levels were measured by using ELISA. IR was evaluated by the HOMA model 

index (HOMA-IR). We found that serum concentrations of leptin and RBP4 

were elevated and adiponectin decreased in overweight patients compared 

to lean controls (P


Obesity 

POSTERS 

different metabolic conditions can induce signifi cantly divergent exhaled gas 

profi les. Integrations of this information into condition-specifi c predictive 

models (for instance use of different gases in predictive algorithms for 

glycemia in obese, T1DM and T2DM) is likely to considerably improve 

accuracy of breath based measurements of plasma variables, accelerating 

the development of portable, clinically usable breath testing devices. 

Supported by: NIH 1UL1RR031985, K24 DK085223 ADA-Funded Research 

1889-P 

Early Aggressive Weight Loss Efforts Using Adjustable Gastric Banding 

Leads to Improvement or “Remission” of Type 2 Diabetes Mellitus 

TED OKERSON, MICHAEL OEFELEIN, SUNIL BHOYRUL, PAMELA BARNETT, JOHN 

B. DIXON, APEX, Irvine, CA, La Jolla, CA, Melbourne, Australia 

Bariatric surgery (malabsorptive or restrictive techniques) has been 

established as an effective treatment to reduce weight in severely obese 

patients. This study reports the 1 year “remission” (elimination of hypoglycemic 

medication) and/or improvement (reduction in hypoglycemic 

medication) of type 2 diabetes mellitus (T2D) after laparoscopic placement 

of an adjustable gastric band (AGB) as documented by T2D medication 

reduction/discontinuation, and the accompanying change in BMI and comorbidities. 

The APEX study is an ongoing 5-year, prospective, multi-center, 

open-label, observational study to assess weight reduction, co-morbidities 

and quality of life after implantation of the LAP-BAND AP ® gastric band, a 

restrictive weight loss technique. This is an interim analysis of subjects who 

reported daily medical therapy for T2D before AGB and who have completed 

the 1 year post-operative scheduled visit. At baseline (BL), 94 out of 436 

subjects (22%) reported T2D requiring daily medical therapy; data from 64 

subjects contained suffi cient information to assess outcome at 48 weeks. 

Overall, 86% had remission and/or improvement in T2D, with remission more 

likely to occur in patients treated earlier after the diagnosis of T2D: 

Remission Improvement Stable Worse 

%(n) 34 (22) 52 (33) 13 (8) 2 (1) 

Mean Duration T2D(mo) 63 76 90 1 

BL BMI 46 44 52 47 

Δ BMI -8.9 -7.6 -8.1 -2.9 

Δ Wt (lb) -55 -48 -52 -18 

% Wt Δ -19 -21 -15 -6 

Baseline BMI, reductions in BMI and % change in weight were not 

statistically different among the groups, although numbers were small. 

As in patients with T2D, resolution or improvement also occurred in other 

pre-existing co-morbidities measured: hypertension (78%), hyperlipidemia 

(57%), depression (71%), obstructive sleep apnea (69%) and GERD (93%). 

These data suggest that a minimally-invasive restrictive gastric banding 

procedure in obese patients with T2D results in clinically meaningful weight 

loss, as well as a reduction in T2D medication requirements, with an earlier 



CATEGORY 

A510 

aggressive weight loss intervention being more likely to facilitate remission 

of disease. Larger and longer-term studies are required. 

Supported by: Allergan 

1890-P 

Ectopic Fat Accumulation and Metabolic Syndrome (MSYN) Are Associated 

with Reduced Subcutaneous Adipose Tissue (SAT) Lipid Storage 

in Obese Postmenopausal Women (OPW) 

MONICA C. SERRA, ALICE S. RYAN, RONALD L. PRIGEON, ANDREW P. GOLDBERG, 

Baltimore, MD 

Impaired expandability of SAT and the accumulation of fat in visceral 

adipose tissue (VAT) and muscle (Low Density Lean Tissue [LDLT]) may lead to 

metabolic dysfunction in obesity. We hypothesize that a decreased ability to 

store triglycerides (TG) in SAT, due to low adipose lipoprotein lipase activity 

(LPL), results in lipid overfl ow into VAT and LDLT stores, insulin resistance (IR 

[measured by HOMA-IR]) and MSYN. To test this hypothesis we measured 

body composition (DXA, CT), MSYN variables, fat cell weight (FCW) and 

LPL in abdominal and gluteal adipose biopsies in 125 Caucasian overweight 

and OPW (25-44 kg/m 2 ; age 45-77 yrs) grouped into lowest and highest 

quintiles of VAT/total abdominal fat (VAT/[VAT+SAT]; mean±SD: 0.17±0.02 

vs. 0.37±0.05). OPW in the highest quintile had a greater prevalence of 

impaired glucose tolerance (IGT; 67% vs. 20%, P

in the pathogenesis of T2D, and suggest that altering the intestinal site of 

delivery of ingested nutrients has therapeutic effects. 

Before surgery 6 M 9 M 12 M 

Weight (kg) 79.4±11.9 74.7±10.9* 75.5±11.2* 75.9±12.4* 

HOMA-IR 5.2±3.1 5.7±3.2 5.6±2.6 6.4±3.3 

Medication score 1.40±0.55 0.89±0.65* 1.04±0.73* 1.05±0.76* 

HbA1c (%) 9.3±1.7 7.0±1.0* 7.1±1.2* 7.4±1.7* 

GLU iAUC (mg/dLx120min) 16300±4600 13700±4400* 12500±3900* 11700±4000* 

C-PEP iAUC (µg/mLx120min) 158±62 306±66* 376±112* 363±90* 

INS iAUC (µU/mLx120min) 882±360 1560±700* 1540±620* 163±610* 

C-PEP secretory response 

(c-PEP iAUC/GLU iAUC) 

0.011±0.008 0.026±0.019* 0.035±0.021* 0.035±0.014* 

Insulinogenic index 

(INS iAUC/GLU iAUC) 

Values are means±SD. 

0.06±0.04 0.13±0.09* 0.16±0.13* 0.15±0.07* 

Value different from before surgery: * p


Obesity 

POSTERS 

1895-P 

Effects of a Three-Month Combined Exercise Program on Fibroblast 

Growth Factor 21 and Fetuin-A Levels and Arterial Stiffness in Obese 

Women 

HYE JIN YOO, HO CHEOL HONG, HAE YOON CHOI, MYONG JIN CHO, YOON JUNG 

KIM, JOO HYUNG KIM, CHAI RYOUNG EUN, SAE JEONG YANG, HEE YOUNG KIM, 

JI A. SEO, NAN HEE KIM, SIN GON KIM, SEI HYUN BAIK, DONG SEOP CHOI, KYUNG 

MOOK CHOI, Seoul, Republic of Korea 

We examined the relationship between brachial-ankle pulse wave velocity 

(baPWV) refl ecting arterial stiffness and the levels of novel hepatokines 

fi broblast growth factor 21 (FGF21) and fetuin-A. In addition, we evaluated the 

effect of a three-month combined aerobic and resistance exercise program on 

FGF21 and fetuin-A levels as well as arterial stiffness in obese women. 

Forty non-diabetic, obese women (BMI = 27.6 ± 2.4 kg/m 2 ) were included 

in the study and were compared before and after a three-month exercise 

program, which was composed of 45 minutes of aerobic exercise at an 

intensity of 60–75 % of the age-predicted maximum heart rate and 20 

minutes of resistance training fi ve times a week. 

At baseline, baPWV levels were signifi cantly correlated with age, BMI, 

systolic blood pressure, high-density lipoprotein cholesterol, fasting glucose, 

and serum FGF21 levels. In a multiple stepwise regression analysis, baPWV 

levels were independently associated with age, BMI, SBP, FGF21, and fetuin-A 

levels (R 2 = 0.744). After the exercise program, BMI, waist circumference, 

SBP, DBP, and triglyceride levels were signifi cantly decreased. Moreover, 

baPWV values were signifi cantly improved (P < 0.001) while FGF21 levels 

declined signifi cantly (P = 0.043). However, fetuin-A levels were not changed 

signifi cantly (P = 0.202). 

A three-month combined exercise program signifi cantly decreased the 

arterial stiffness and FGF21 levels in obese Korean women. 

1896-P 

Five-Year Follow-Up of Type 2 Diabetic Patients Who Underwent Bariatric 

Surgery 

HELEN M. HENEGHAN, FADY MOUSTARAH, SHAI MERON-ELDAR, SANGEETA 

KASHYAP, JOHN KIRWAN, BIPAN CHAND, STACY BRETHAUER, TOMAZ ROGULA, 

MATTHEW KROH, LAURENCE KENNEDY, PHILP R. SCHAUER, Cleveland, OH 

In addition to weight loss, bariatric surgery has powerful metabolic 

effects, including resolution of obesity-related comorbidities such as type 

2 diabetes mellitus (T2DM). However, the durability of these benefi ts is 

largely unknown. The aim of this study was to determine 5-year outcomes 

of morbidly obese diabetic patients who underwent bariatric surgery, and to 

identify factors associated with durable diabetes remission. We identifi ed 

all T2DM patients who underwent bariatric surgery at our institution, and 



CATEGORY 

A512 

had 5-year follow-up data available. Patient’s current T2DM status was 

determined by biochemical analyses and review of medications. Remission 

was defi ned as fasting glucose 0.05), 

whereas associations were detected for VAT/SAT ratio with LYPLAL1, GRB14, 

and ADAMTS9 (p-value range 0.0016-0.03); these same SNPs were also 

associated with SAT (p

1898-P 

Heritability of the Risk Factors Characteristic for the Metabolic 

Syndrome: A Twin Study 

GYÖRGY JERMENDY, LEVENTE LITTVAY, RITA STEINBACH, ÁDÁM JERMENDY, 

JÁNOS OSZTOVITS, Budapest, Hungary 

Both genetic and environmental factors play role in the pathogenesis of the 

metabolic syndrome and obesity. The precise magnitude of genetic infl uence 

on the components of the metabolic syndrome is poorly described. 

In our study, a total of 101 (63 monozygotic [MZ] and 38 dizygotic [DZ]) 

adult twin pairs (n=202; mean age: 43.3±15.8 years) were investigated. 

Past medical history was recorded and physical examination was carried 

out in each subject. Fasting venous blood samples were used for measuring 

laboratory parameters. Intraclass correlations were evaluated and 

heritability model analyses (A-C-E model) were performed. All parameters 

were corrected for age, gender and BMI values. 

The intraclass correlation for waist circumference was higher in MZ 

(r=0.744; p


Obesity 

POSTERS 

1902-P 

Intentional Weight Loss Is Associated with Increased Total and 

High Molecular Weight Adiponectin in Men and Women with Type 

2 Diabetes in Look AHEAD (Action for Health in Diabetes); Findings 

on Fitness and Adiposity Change 

L. MARIA BELALCAZAR, STEVEN M. HAFFNER, WEI LANG, RON C. HOOGEVEEN, 

KATHERINE M. DONADIO, DAWN C. SCHWENKE, F. XAVIER PI-SUNYER, RUSSELL 

P. TRACY, ANDREA M. KRISKA, CHRISTIE M. BALLANTYNE, Galveston, TX, Houston, 

TX, Winston-Salem, NC, Phoenix, AZ, New York, NY, Burlington, VT, Pittsburgh, PA 

Adiponectin and particularly high molecular weight adiponectin (HMW-A) 

levels are low in people with type 2 diabetes. Little is known about the 

differential effects of weight loss and increased fi tness on total and HMW-A 

in obese diabetic persons. In a subset of 1,397 Look AHEAD participants (with 

characteristics similar to the whole sample) we evaluated the effects of a 

1-year intensive lifestyle intervention for weight loss (ILI) and of usual care 

(Diabetes Support and Educations [DSE]) on total and HMW-A levels using a 

quantitative ELISA and investigated the independent contribution of adiposity 

and fi tness changes on their levels. Look AHEAD is a randomized trial evaluating 

whether ILI will reduce cardiovascular events/mortality when compared to 

DSE in obese type 2 diabetic persons. Subjects (mean age 57.2 years; BMI 

36.3 kg/m 2 ; submaximal fi tness 5.2 METS) had baseline total and HMW-A 

levels (median [interquartile range]) of 4.1 (3.0, 5.8) and 1.6 (1.0, 2.7) in men 

and 5.1 (3.8, 7.5) and 2.2 (1.4, 3.5) µg/mL in women, respectively. At 1-year, ILI 

participants lost 8% of baseline weight and increased fi tness by 21% (vs 0.6% 

and 6% in DSE, respectively). ILI increased baseline total adiponectin by 24% 

in men and 6% in women and HMW-A by 36% and 15% in men and women, 

respectively (p

1906-P 

Long-Term Weight Loss and Improvement in Glycemic Parameters 

with Low-Dose, Controlled-Release Phentermine/Topiramate (PHEN/ 

TPM CR) 

ROBERT F. KUSHNER, W. TIMOTHY GARVEY, BARBARA TROUPIN, WESLEY W. 

DAY, Chicago, IL, Birmingham, AL, Mountain View, CA 

Obesity is an epidemic linked to chronic comorbidities, including type 2 

diabetes mellitus (T2DM). Previously, weight loss associated with PHEN/ 

TPM CR demonstrated improvements in glycemic parameters through 56 

weeks in the CONQUER study. SEQUEL, an extension of CONQUER, was 

conducted to evaluate long-term (108 weeks) effects of PHEN/TPM CR on 

weight loss and glycemic parameters related to T2DM. This double-blind 

extension maintained the original blinded treatment groups for an additional 

52 weeks in subjects completing 56 weeks of CONQUER: placebo (PBO; 

n=227), PHEN 7.5 mg/TPM CR 46 mg (7.5/46; n=153), or PHEN 15 mg/TPM 

CR 92 mg (15/92; n=295). All subjects with T2DM were managed to the ADA 

standards of care. In the overall ITT-LOCF sample, least-squares (LS) mean 

percent weight loss was signifi cantly greater with both doses of PHEN/TPM 

CR compared with PBO (P


Obesity 

POSTERS 

The multiple adjusted odds ratio for those who had the lowest creatinine 

level (≤0.80mg/dL) was 1.785 (95% CI 1.145– 2.781) compared with those 

who had the highest creatinine level (1.01–1.20). In conclusion, lower serum 

creatinine was associated with an increased risk of glucose abnormality, but 

not other components of MS among obese Korean men. 

1910-P 

Metabolic Syndrome Is Associated with Impaired Pulmonary Function 

in a Restrictive, but Not Obstructive, Pattern, in Japanese Men 

TAKEHIDE OGIHARA, NORIKO CHIKATSU, TAKESHI NAWA, RYOICHI NAMBA, 

YUSUKE YAMAMOTO, MICHIKO MURAOSA, SHINJI HIRAI, YUJI OKA, Hitachi, Japan 

Metabolic syndrome (MS) has been shown to be associated with impaired 

pulmonary function, especially chronic obstructive pulmonary disease (COPD). 

Systemic infl ammation is a possible mechanism underlying the association 

between these disorders. In this study, we investigated the effect of MS on 

pulmonary function by analyzing data from 9,573 men aged 22 to 89 years. 

Based on the diagnostic criteria for MS in Japan, current/former-smokers 

and those who never smoked were each divided into MS and non-MS groups. 

Then we examined the relationship between MS and measures of pulmonary 

function (including percentage predicted forced vital capacity; %FVC, 

percentage predicted forced expiratory volume in 1 s; %FEV 1 , and FEV 1 - to- 

FVC ratio). Next, we investigated the correlation between visceral fat area 

and each measure of pulmonary function. Lastly, we analyzed the infl uence of 

MS diagnostic criteria on each measure of pulmonary function using stepwise 

regression analysis. Non-smokers and smokers with MS showed signifi cantly 

lower %FVC and %FEV 1 than those without MS, respectively. However, 

FEV 1 - to-FVC ratio was signifi cantly higher in subjects with MS than in those 

without MS. Among smokers and non-smokers, visceral fat area showed weak 

negative correlations with %FVC and %FEV 1 . In the regression equation with 

%FVC as a dependent variable, waist circumference, blood pressure, lipids 

and fasting blood glucose were signifi cant negative explanatory variables. 

However, in that with FEV 1 - to-FVC ratio, lipids and fasting blood glucose were 

positive explanatory variables. Thus, indicators of impaired pulmonary function 

associated with MS are %FVC and %FEV 1 , suggesting that MS is associated 

with impaired pulmonary function in a restrictive, but not obstructive, pattern. 

As airfl ow limitation for the diagnosis of COPD is judged by FEV 1 - to-FVC ratio, 

there is a risk of missing early COPD among smokers with MS. Therefore, 

smokers with MS should be advised early of the need for smoking cessation. 

1911-P 

Metabolically Healthy Obese Individuals Have Decreased Heart 

Failure Risk Compared to Normal-Weight People in a Six-Year Mediter 

ranean Study 

VASILLIS VOULGARIS, CHRISTINA VOULGARI, NICHOLAS TENTOLOURIS, PAUL 

POIRIER, Athens, Greece, Quebec, QC, Canada 

Heart failure (HF) is one of the leading causes of mortality and its prevalence 

continues to rise, despite the decline in cardiovascular death rates. Among 

the risk factors identifi ed as consistently associated with its development 

are Obesity and Metabolic Syndrome (MetS). “Metabolically Healthy Obese” 

(MHO) people, despite their body fat, display a favorable metabolic profi le 

characterized by high levels of insulin-sensitivity, and a benefi cial bloodpressure, 

glycemic, lipids, and infl ammation profi le. It remains controversial 

whether this healthier metabolic profi le is translated into a lower cardiovascular 

risk compared with normal-weight individuals with MetS. 

A total of 550 individuals without diabetes or baseline macrovascular 

complications were studied. Participants were classifi ed by presence 

(n=271) or absence (n=279) of MetS and by BMI (BMI;

There is an association of the Gly482Ser polymorphism with type 2 

diabetes and obesity. No previous study has evaluated the association of 

this polymorphism with the outcomes of bariatric surgery. We tested the 

hypothesis that Gly482Ser polymorphism could predict clinical, laboratorial 

and structural atherosclerotic marker [carotid intima-media thickness 

(C-IMT)] outomes. Patients (n=55) were submitted to Roux-en-Y gastric 

bypass (RYGB) and evaluated for anthropometric, C-IMT, metabolic and 

infl ammatory parameters at baseline, 1, 6 and 12 mo after RYGB. The 

polymerase chain reaction- restriction fragment length polymorphism 

was performed. Serum IL-6, adiponectin, leptin, insulin and MCP-1 were 

determined. An euglycemic-hyperinsulinemic clamp was performed. C-IMT 

was measured 1.0 cm distal to the bulbus over a length of 1.0 cm of carotid 

arteries. At baseline, all parameters were similar between groups. At 1-year 

after surgery, BMI dropped from 45 to 28 in the Gly/Gly group, and from 44 to 

28 in Gly/Ser+Ser/Ser group. There was a signifi cant reduction in the waist/ 

hip ratio only in the Gly/Ser+Ser/Ser group. Lipid profi le improved after 12 

mo similarly in both groups. Fasting blood glucose dropped signifi cantly only 

in the Gly/Ser+Ser/Ser. Insulin, HOMA-IR and clamp results, and adipokines 

(leptin, adiponectin) presented signifi cant changes similarly in both groups. 

Both Gly/Gly and Gly/Ser+Ser/Ser groups presented reductions of us-CRP, 

MCP-1 and IL-6. Signifi cant inter-group differences were detected for us-CRP, 

blood leukocyte counts and IL-6. Both Gly/Gly and Gly/Ser+Ser/Ser patients 

presented signifi cant reductions at 12 mo (0.20 mm, and 0.27 mm, respectively, 

p


Obesity 

POSTERS 

1918-P 

Prevalence-Adjusted Cost of Comorbidities in Overweight/Obese 

Patients with a Body Mass Index 25-34.9 vs ≥35 

DIANA I. BRIXNER, MORGAN BRON, BRANDON BELLOWS, XIANGYANG YE, 

VENKATESH HARIKRISHNAN, GARY M. ODERDA, Salt Lake City, UT, Deerfi eld, IL 

Obesity is associated with an increased prevalence of comorbidities. 

This study looked at prevalence and cost of comorbidities in 2 categories of 

overweight/obese patients based on BMI. 

Prevalence of 25 comorbidities in patients with BMI 25-34.9 and ≥35 was 

calculated and ranked based on data from the GE Centricity EMR research 

database. Patients whose BMI was within NHLBI guideline categories for 

overweight (25-29.9), class I obesity (30-34.9) and class II or III obesity (≥35) 

between 3/1/05 and 6/30/09 were included. BMI index date was defi ned as 

date of either only BMI on record, or latest BMI for patients with 2 or more 

BMI readings. Prevalence of comorbidities was based on presence of ICD- 

9 code within 2 y prior to BMI index date. Annual comorbidity-associated 

costs were based on an ICD-9 code for overweight or obesity and patient 

activity for 1 y post comorbidity index date in Thomson Reuters MedStat 

MarketScan database. Prevalence-adjusted cost of comorbidities was 

determined by multiplying annual comorbidity-associated cost per patient 

from MarketScan by comorbidity prevalence rate ratio from 2 BMI groups 

of the EMR database. 

The GE EMR database identifi ed 109,885 patients with BMI ≥25 and 2 y EMR 

activity before BMI index date. The 3 comorbidities with highest prevalence (BMI 

25-34.9, ≥35) were hyperlipidemia (14.6%, 16.3%), hypertension (10.2%, 17.7%), 

and back pain (7.5%, 8.3%). Annual comorbidity-associated cost per overweight/ 

obese patient was $1136 for hyperlipidemia, $1511 for hypertension, and $2081 

for back pain. Prevalence-adjusted per patient cost increased from BMI 25-34.9 

to BMI ≥35 in hyperlipidemia ($166 to $185), hypertension ($154 to $268), and 

back pain ($156 to $173). As an example, a plan with 1,000 members with BMI 

25-34.9 would have hyperlipidemia-associated cost of $166,000 vs $185,000 for 

a plan with 1,000 members with BMI ≥35. 

This study confi rmed the incremental impact of weight on prevalence and 

cost of selected comorbidities in overweight or obese patients. Management 

or therapies that can lower patients’ BMI have the potential to decrease 

comorbidity rates and may lower costs for health plans. 

1919-P 

Short-Term Evolution of the Insulin Resistance Depending on the 

Type of Bariatric Surgery 

LOURDES GARRIDO-SANCHEZ, MORA MURRI, JOSE RIVAS-BECERRA, DIEGO 

FERNANDEZ-GARCIA, JUAN ALCAIDE, EDUARDO GARCIA-FUENTES, FRANCISCO 

TINAHONES, Tarragona, Spain, Malaga, Spain 

Obesity is very often accompanied by other diseases, the most common being 

type 2 diabetes mellitus and cardiovascular complications. Bariatric surgery 

is almost the only effective strategy for treating morbidly obese patients. 

The aims of this study was evaluate the metabolic changes that occur in the 

early stage after two types of bariatric surgery, biliopancreatic diversión of 

Scopinaro (BPD) and Sleeve gastrectomy (SG), in morbidly obese persons. The 

study was undertaken in 31 morbidly obese persons (7 men and 24 women) 15 

days before and 15, 30, 45 y 90 days after bariatric surgery (times 0, 1, 2, 3, and 

4, respectively). There is a signifi cant improvement in anthropometric variables 

as result of the two types of bariatric surgery, without signifi cant differences 

between both types of interventions. In patients undergoing BPD, serum 

glucose, cholesterol, triglycerides, HDL-cholesterol and FFA were signifi cantly 

reduced. The changes that occur in these biochemical variables following the 

SG were not signifi cant. Insulin resistance decreased signifi cantly over the 

90 days after surgery, showing the highest decrease at 15 days. Meanwhile, 

in patients undergoing SG, insulin resistance worsened at 15 days and later 

diminished. In conclusion, this study shows that the surgical technique that 

excludes the duodenum (BPD) has immediate postoperative changes in the 

degree of insulin resistance in morbidly obese patients compared to those 

techniques that do not exclude (SG). 

Supported by: JCI-2009-04086, CP04/00133, PS09/01060, PS09/00997 

1920-P 

Small Decrements in Glucose Increase Hunger in the Presence of 

Visual Food Cues 

RENATA BELFORT DE AGUIAR, SARITA NAIK, DONGJU SEO, RAJITA SINHA, 

ROBERT SHERWIN, New Haven, CT 

Hypoglycemia is associated with increased hunger, triggering increased 

food consumption. The mechanism for how circulating glucose infl uence 

food-seeking behavior is unclear. In this study, we evaluated the effects of 

small changes in glucose within the physiological range on measurements 



CATEGORY 

A518 

of food motivation in response to visual food cues. Ten healthy subjects 

(mean age 31±8, A1c 5.4±0.4) underwent a 2mU/kg.min hyperinsulinemic 

euglycemic clamp (goal=plasma glucose ∼90mg/dl) while viewing high fat, 

low fat food and non-food pictures, 2 hours after receiving a standardized 

meal. Liking and wanting ratings were collected for each picture. Hunger 

ratings were collected at baseline, 60, 85min and at 2 hrs. Initially plasma 

glucose rose from 83±9 mg/dL at baseline to 98±11 mg/dL after 10mins. 

During the interval between 10 and 60 min plasma glucose showed more 

variability, although mean levels decreased slightly to 93±7mg/dL (it 

decreased by 14+/-4 mg/dL in 6 subjects and increased by 11+/-5 mg/dL in 

4 subjects). Subsequently, plasma glucose stabilized at 95±9mg/dL until the 

end of the clamp. Hunger ratings, after subjects viewed food and non-food 

images, signifi cantly increased from 1.7±0.7 at baseline to 4.0±0.8 at 60min 

(p=0.01) and 5.2±2.9 at 2 hrs. (p=0.004). 

Hunger levels at the end of the clamp were associated with glucose at 

10min (r=0.816,p=0.004) and 60min (r=-0.702,p=0.02) and the drop in glucose 

between 10 and 60mins (r=0.848,p=0.002). The changes in glucose were not 

associated with signifi cant increases in plasma glucagon, cortisol, FFA, or 

ghrelin from baseline levels. In addition, leptin levels increased (p

Several studies indicated that obese subjects, particularly those with 

visceral fat accumulation, have reduced plasma levels of adiponectin. Other 

results indicate that rs1049353 (1359G/A) polymorphism of the CNR1 gene is 

associated with increased abdominal circumference (AC). 

The aim of our study was to investigate the association between the 

rs1049353 polymorphism and low adiponectin levels in subjects with 

abdominal obesity (AO). 

The study included 377 subjects divided in: 219 subjects with AO 

(AC≥102cm in men; AC≥88cm in women) and 158 subjects without AO. The 

groups were divided in 2 subgroups: one with low adiponectin levels (


Obesity 

POSTERS 

within an intron of DDX60L which encodes a ATP-dependent helicase 

binding with RNA. CD83 and RNF182 are the only 2 genes within ±500kb 

of rs6459353. CD83 is a cell-surface glycoprotein that regulates immunity 

response, and RNF182 is a brain-enriched E3 ubiquitin ligase that may 

participate in controlling the neurotransmitter release mechanism. To 

explore potential associations between these 3 genes and BMI, the putative 

promoter regions and all exons and exon-intron boundaries of these genes 

were sequenced in 24 Pima Indians. 4 novel SNPs were identifi ed (2 in 

DDX60L, 1 in CD83 and 1 in RNF182); one novel SNP within DDX60L predicts a 

1323F/I substitution and the other 3 SNPs are in untranslated regions. These 

SNPs, in addition to 53 tag SNPs from the HapMap (CHB) are currently being 

genotyped in our samples. 

1926-P 

Vitamin D Insuffi ciency, a Risk Factor for Prediabetes and Diabetes 

Type 2 in Abdominal Obesity Women 

TATIANA KARONOVA, ELENA MICHEEVA, DANY AZZI, ELENA KONOPLYANNI- 

KOVA, IDELYA MAMINA, ELENA KRASILNIKOVA, EUGENIYA PATRAKEEVA, 

ELENA GRINEVA, EUGENIY SHLYAKHTO, St. Petersburg, Russia 

Some studies suggest that serum 25(OH) vitamin D concentration is 

inversely associated with insulin resistance and the prevalence of diabetes 

type 2. However due to the uncertainty of these associations, we examined 

the relationship between serum concentration of 25(OH)D and glucose as well 

as insulin levels, HOMA-IR, and ISI (0,120) in abdominal obesity women. 

We examined 162 women 40 to 52 years old (mean age 46.9±5.5) with 

abdominal obesity (according to IDF criteria, 2005). The mean waist circumference 

was 96.5±1.7 cm, body mass index (BMI) was 31.3±0.4 kg/m 2 , waist-to-hip ratio 

(WHR) - 0.9±0.004. Serum 25(OH)D and insulin levels were determined by 

ELISA, plasma glucose levels - by standard biochemistry. All patients underwent 

standard oral glucose-tolerance test (75 g of glucose). Insulin resistance was 

evaluated by HOMA-IR and insulin sensitivity by ISI (0, 120). 

Glucose level was 6.1±0.2 mMol/L, insulin level 14.9±0.9 (mIU/ml), HOMA- 

IR - 3.2±0.1, ISI (0, 120) – 7.52±0.39. Serum 25(OH)D level was from 19.4 to 

134.1 nMol/L (mean 55.6±2.9). Vitamin D insuffi ciency and defi ciency was 

revealed in 83.6% of patients, only 16.4% women had normal 25(OH)D level. 

Glucose intolerance was revealed in 44 women (25.9%), diabetes type 2 

was diagnosed in 25 women (16.1%). Correlation analysis showed inversed 

correlations between serum 25 (OH)D level and fasting glucose, insulin 

levels (r=0.14, p=0.05; r=0.20, p=0.008 respectively) and ISI (0,120) (r=0.33, 

p=0.001). We found that HOMA-IR was related to BMI value (r=0.5, p=0.001), 

but not with 25(OH)D levels (r=-0.14, p=0.08). 

Our results showed that women with abdominal obesity have higher 

tendency for developing vitamin D defi ciency and this later is correlated with 

high fasting plasma glucose, serum insulin levels and decreased tissues 

insulin sensitivity. Hence vitamin D insuffi ciency might possibly be a risk 

factor for pre diabetes and diabetes type 2. 

1927-P 

Vitamin D-Associated Polymorphisms Are Related to Insulin Resistance 

and Vitamin D Defi ciency in Polycystic Ovary Syndrome 

WITHDRAWN 

ELISABETH WEHR, THOMAS R. PIEBER, BARBARA OBERMAYER-PIETSCH, Graz, 

Austria 

Women with polycystic ovary syndrome (PCOS) frequently suffer from 

insulin resistance, which might be related to vitamin D metabolism. We 

aimed to investigate the association of polymorphisms in the vitamin D 

receptor (VDR) gene and vitamin D level associated genes with metabolic 

and endocrine parameters in PCOS and control women. 

Metabolic, endocrine, and anthropometric measurements and oral 

glucose tolerance tests were done in 545 PCOS and 145 control women. 

Genotyping of VDR (Cdx-2, Bsm-I, Fok-I, Apa-I, and Taq-I), GC, DHCR7, and 

CYP2R1 polymorphisms was performed. 

25-hydroxyvitamin D (25[OH]D) levels correlated negative with insulin 

resistance and positive with insulin sensitivity (p

subjects completing 56 weeks of treatment, NB was generally well tolerated 

and associated with dose-dependent sustained weight loss and numerous 

improvements in weight-related risk factors, including lipids and glycemic 

parameters, as well as improved quality of life and control of eating. 

Supported by: Orexigen Therapeutics 


ISLET BIOLOGY—APOPTOSIS 

[See also: Presidents Posters 469-PP to 470-PP, page A130.] 

Guided Audio Tour: Apoptosis (Posters 1929-P to 1937-P), see page 11. 

& 1929-P 

Impact of Vildagliptin on Glucose Tolerance, and beta Cell Mass in 

Insulin Receptor Substrate-2-Knockout Mice 

KOICHIRO SATO, AKINOBU NAKAMURA, JUN SHIRAKAWA, TOMONORI MURA- 

OKA, YU TOGASHI, KAZUKI ORIME, YASUO TERAUCHI, Yokohama, Japan 

Loss of beta-cell mass underlies the development of diabetes and insulin 

receptor substrate-2 (Irs2) is critically required for beta cell growth and 

survival. GLP-1 reportedly increased islet proliferation and reduced apoptosis 

of beta-cells in rodents. In this study, we explored the chronic effect of 

dipeptidyl peptidase-4 (DPP-4) inhibitor vildagliptin on glucose tolerance 

and beta-cell mass in Irs2-knockout (Irs2-/- ) mice fed a high-fat diet for 20 

weeks. Wild type (WT) mice and Irs2-/- mice of the same genetic background 

were fed standard-chow until 8 weeks of age, and then given free access 

to high-fat diet for 20 weeks. Half of the animals in each genotype were 

given vildagliptin in the drinking water (0.3 mg/ml) and others were given 

tap water without vildagliptin. Whereas there were no differences in body 

weight or food intake, vildagliptin signifi cantly reduced fasting blood glucose 

in Irs2-/- mice. In both genotypes of mice, vildagliptin signifi cantly decreased 

AUC0-120min blood glucose and signifi cantly increased the insulin response to 

glucose during oral glucose tolerance test (OGTT). To evaluate the chronic 

effect rather than the direct effect, we also performed OGTT one day after 

discontinuation of vildagliptin administration. AUC0-120min blood glucose was 

still signifi cantly decreased and the insulin response to glucose during OGTT 

was signifi cantly increased in Irs2-/- mice treated with vildagliptin compared 

with those without vildagliptin. Histochemical analysis of pancreatic islets 

revealed that treatment with vildagliptin signifi cantly increased beta-cell 

mass (p

2011 ADA Posters 1261-2041.indd - Diabetes

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