INTEGRATED PHYSIOLOGY—INSULINCATEGORY SECRETION IN VIVO

tigate this, we built upon our zinc data to explore activation of IRAP, via An- giotensin IV, in our rodent model of T2DM. The present data show that zinc is Measure (Mean ± SE) South Asian Caucasian P a neuromodulator, regulating hippocampal memory processes, and suggest Clamp M (mg/kg·min) 4.48±0.24 7.46±0.34 < 0.0001 that zinc supplementation and administration of zinc-dependent peptides Clamp Si (mg/kg·min per μU/ml) 0.088±0.009 0.151±0.009 < 0.0001 may be useful therapeutic agents for cognitive impairment in T2DM. Matsuda Index 7.60±0.99 13.60±1.79 0.0036 Supported by: NIH and the Alzheimer’s Association HOMA-IR (μU/ml·mmol/l) 1.56±0.19 0.77±0.07 0.0013 2762-PO OGTT Mean Insulin (μU/ml) 49.1±6.2 26.5±4.1 0.0064 Tumor Necrosis Factor-ƴ-Induced NF-ƽB Signaling in Human Pan- OGTT Mean Glucose (mg/dl) 115±6 107±5 NS creatic Islets and Human Ƶ-Cells JAI PARKASH, Miami, FL 2+ TNF-_ induces dysregulation of intracellular calcium, [Ca ]i, in `-cells by 2+ 2764-PO decreasing the levels of a cytoplasmic Ca binding protein calbindin-D28k. In the present study we wanted to test the hypothesis that “In human islets Fatty Acids Stimulate Glucose-Induced Insulin Secretion In Vivo in and human islets cells including human pancreatic `-cells, TNF-_ activates Mice NF-gB signaling pathway resulting in nuclear translocation of NF-gB and PAYAL R. PATEL, HWI JIN KO, DAE YOUNG JUNG, XIAODI HU, ISAAC SHIELDS, transcriptional activation by NF-gB”. To test this hypothesis, we treated hu- SARAH E. KINICKI, ASHWINI D. JAVLEKAR, ANNIE H. HUYLER, JUN HO LEE, YUN man islets and human islets cells including human pancreatic `-cells with HEE NOH, JASON K. KIM, Worcester, MA, Seoul, Republic of Korea increasing concentrations of TNF-_. Upon treatment of human islets and Chronic hyperinsulinemia is a compensatory event when insulin resis- human islets cells including human pancreatic `-cells with 2, 5, 10, 20, and tance develops in obesity. Increasing evidence suggests that hyperplasia 30 ng/ml TNF-_, the relative increases in the transcriptional activities by and hypertrophy of pancreatic islets and associated hyper-secretion of in- NF-gB, determined by utilizing the DNA binding activity of NF-gB in an ELI- sulin are precipitating events to islet decompensation, `-cell apoptosis, and SA-based kit, were 1.69±0.32, 1.42± 0.07, 1.77± 0.43, 2.02± 0.02, and 1.96± type 2 diabetes. Thus, it is important to understand the molecular mecha- 0.2 folds respectively (n=3, P< 0.05). The nuclear translocation of NF-gB nism by which obesity leads to chronic hyperinsulinemia. Here we exam- measured by immunofl uorescence confocal microscopy showed the nuclear ined the effects of fatty acids on islet function in vivo. Male C57BL/6 mice translocation of NF-gB both in human islets and in human islet cells includ- received an intravenous infusion of lipid emulsion at 2 different doses (2.5 ing human pancreatic `-cells. These observations suggest that in human or 5 ml/hr/kg; n=7-8/group) or glycerol (controls; n=8) for 5 hrs. Lipid infusion islets and in human islet cells including human pancreatic `-cells, the TNF-_ increased circulating fatty acids (FA) levels to 3~4 mM in a dose-dependent indeed activates NF-gB signaling pathway resulting in nuclear transloca- manner, whereas a glycerol infusion maintained FAs near basal levels (~1 tion of NF-gB and transcriptional activation by NF-gB. Although the exact mM) (Fig. 1; *p<0.05 vs. glycerol). After the 5 hr lipid infusion, a 2-hr hyper- mechanism of NF-gB activation by Ca2+ is still not known, what is known is glycemic clamp was conducted to assess glucose-induced insulin secretion that Ca2+ signaling in cells is generally presented as changes in Ca2+ concen- in awake mice. Plasma glucose levels were quickly raised and maintained at tration as brief spikes which are often organized as regenerative Ca2+ waves. ~20 mM. Hyperglycemia caused a rapid and sustained increase in plasma The intracellular Ca2+ waves can induce changes in the gene expression and insulin levels (Fig. 2). Lipid infusion at 2.5 and 5 ml/hr/kg induced further thus regulate several signal-transduction pathways. dose-dependent increases in plasma insulin levels (Fig. 2). Area-under-curve Supported by: NIH/NIGMS Grant to J.P., SC3GM084751 of plasma insulin levels was signifi cantly increased by 20~40% compared to glycerol (Fig. 3). Overall, these results indicate that elevated circulating FAs dose-dependently increase insulin secretion in mice. Our fi ndings further suggest that fatty acids potently regulate insulin secretion and contribute to chronic hyperinsulinemia in obesity. INTEGRATED PHYSIOLOGY—INSULIN SECRETION IN VIVO

2763-PO Comparison of Measures of Insulin Sensitivity in South Asians SUBBULAXMI TRIKUDANATHAN, ANNASWAMY RAJI, BINDU CHAMARTHI, EL- LEN W. SEELY, DONALD C. SIMONSON, Boston, MA South Asians have increased visceral adiposity, insulin resistance, and diabetes compared with Caucasians of European origin. Quantifying insu- lin sensitivity in this population is vital for large clinical and epidemiologi- cal studies. Surrogate measures of insulin sensitivity have been validated against hyperinsulinemic euglycemic clamps in Caucasian populations, but ethnicity and variations in body composition may affect these surrogate in- dices. We compared insulin sensitivity determined by the euglycemic clamp (1 mU/kg·min), the Matsuda index (75 gm OGTT) and HOMA-IR in 23 non- diabetic South Asians and 18 Caucasians who had similar age (mean ± SE = 34 ± 2 vs. 36 ± 3 yrs), BMI (25.2 ± 1.1 vs. 24.6 ± 0.9 kg/m²), waist-hip ratio (0.81 ± 0.02 vs. 0.77 ± 0.02), and BP (131 ± 4 / 69 ± 3 vs. 123 ± 4 / 65 ± 3). Fasting metabolic measures showed similar FPG (91 ± 2 vs. 89 ± 2 mg/dl) and Supported by: NIH (R01-DK080756) Obesity LDL-C (107 ± 6 vs. 104 ± 7 mg/dl), but South Asians had lower HDL-C (49 ± 2 vs. 60 ± 4 mg/dl; P = 0.0095), higher triglycerides (112 ± 16 vs. 82 ± 10 mg/ 2765-PO dl), and higher fasting insulin (6.9 ± 0.8 vs. 3.6 ± 0.3 μU/ml; P = 0.0016). All Delayed Insulin Secretory Responses Are Seen in Patients With PUBLISHED ONLY Integrated Physiology/ measures of insulin sensitivity were signifi cantly impaired in South Asians Gout (Table). The Matsuda index was correlated with the clamp M value in South SHENGLI YAN, CUICUI LIANG, YANGANG WANG, Qingdao, China Asians (r = 0.50, p = 0.014), Caucasians (r = 0.47, p = 0.046), and in both groups From our previous data of epidemiological investigation of gout and hy- combined (r = 0.62, p < 0.0001). HOMA-IR also was highly correlated with peruricemia in Shandong coastal area we found that the prevalence of dia- the clamp M in South Asians (r = 0.56; p = 0.005), but not in Caucasians (r = betes in patients with gout and/or hyperuricemia was signifi cantly higher 0.34; p = 0.17). Thus, the Matsuda index and HOMA-IR provide simple and than that of normal population. To explore the change of islet `-cell function valid surrogate measures of insulin sensitivity in South Asians, which may in patients with gout, blood samples of 76 patients with gout(37 gout with be useful in performing larger clinical studies in this population. IGT and /or IFG,39 gout with NGT)and 30 healthy subjects were drawn at 0,2,4 and 6 min for determination of the fi rst phase insulin secretion with intravenous arginine stimulation. In the very next day, the second phase insulin secretion at 0, 60, 120 min was determined with OGTT. Statistical

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A688 INTEGRATED PHYSIOLOGY—INSULINCATEGORY SECRETION IN VIVO analysis was performed using the unpaired T test for comparisons between 2768-PO each group. Correlations between the clinical parameters were evaluated by GLP-1 and Exendin-4 Inhibit Insulinotropic Effects of Glucagon and using multiple step regression analysis. The time of peak distributed differ- Oxyntomodulin in Cattle ently between the gouty group and the healthy group (L-arginine stimulation SINT THANTHAN, HONGQIONG ZHAO, SWE YANNAING, HIDETO KUWAYAMA, 2 min:100% vs 27.6%, P<0.05;OGTT 30-60 min:100% vs 32.9%, P<0.05). The Obihiro, Japan peak value of insulin stimulated by L-arginine and OGTT of gouty group was GLP-1, glucagon and oxyntomodulin (OXM) play an important role in glu- higher than that in the healthy group(P<0.05;P<0.01). The incremental plasma cose homeostasis. GLP-1 and GLP-1 analogs, such as exenatide (a synthetic insulin area under the curves (AUCs) after stimulation of L-arginine and OGTT form of exendin-4 (1-39) amide) and liraglutide, are used for treatment of was higher in gouty group than the healthy group (P<0.01;P<0.01). The time type 2 diabetes in humans. This study was aimed to investigate the rela- of peak signifi cantly delayed after the stimulation of OGTT in the gout with tionships among these 4 peptides in the insulinotropic action in cattle. Four IGT and /or IFG subgroup (P<0.05). Multiple step regression analysis showed mo-old Holstein steers (111 ± 2 kg, n = 8) were assigned randomly in an in- that islet secretion was positively correlated with BMI, waist circumference complete Latin square design (8 steers x 9 treatments x 9 experiment days). and uric acid (P<0.05). We conclude that insulin secretory response both in They were administered with vehicle (0.1 % BSA in saline as a control), GLP-1 the fi rst and the second phase are delayed in patients with gout and that the 0. 25 μg/kg BW, exendin-4 0.25 μg/kg BW, glucagon 2 μg/kg BW, OXM 2 μg/ delayed insulin secretory response in the second phase in gout patients with kg BW, GLP-1 0. 25 μg + glucagon 2 μg/kg BW, GLP-1 0. 25 μg + OXM 2 μg/kg IGT and /or IFG is more obvious than that in gout patients with NGT. BW, exendin-4 0. 25 μg + glucagon 2 μg/kg BW, exendin-4 0. 25 μg + OXM 2 μg/kg BW. Injection of peptide and blood sampling were done via a catheter 2766-PO inserted into the jugular vein of the steers. Plasma insulin concentrations Angiotensin-(1-7) Counter-Regulates the Inhibitory Effect of Angio- were analysed by radioimmunoassay method. Results showed that insulino- tensin II on Akt Phosphorylation in NIT-1 Cells tropic effects of OXM and glucagon were inhibited by GLP-1 and exendin-4. MINGTONG XU, JIANI CHAI, SHENGNENG XUE, LI YAN, Guangzhou, China Our fi ndings expanded the relationships among GLP-1, exendin-4, OXM and Angiotensin II has been demonstrated to exert a deleterious effect on glucagon in the insulinotropic actions. insulin sensitivity. Angiotensin-(1-7) opposes many actions of Ang II. But regulation of AngII,Ang-(1-7)on ` cell insulin signal cascade remains largely unknown. In present study, we evaluated the effect of AngII, Ang-(1-7) on phosphorylation level of signaling molecules in insulin signal pathway in NIT-1 cells. NIT-1 cells were incubated with graded concentrations of AngII 24h: 0, 10-7, 10-6, 10-5, 10-4 mol/L; graded concentrations of Ang-(1-7) 0, 10-7, 10-6, 10-5, 10-4 mol/L. Then cells were divided into control, AngII, Ang-(1-7), AngII+ Ang-(1-7) group. Tyrosine phosphorylation level of insulin receptor ` subunit(IR-`-Tyr), serine phosphorylation level of protein kinase B (Akt-Ser) were detected by Western Blot. Insulin-stimulated IR-`-Tyr and Akt-Ser phosphorylation signifi cantly decreased in AngII10-5 and AngII10-4mol/L group; both insulin-stimulated IR-`-Tyr and Akt-Ser phosphorylation in Ang-(1-7) treatment groups were not signifi cantly different from the control group; interestingly, Ang II-induced inhibition of Akt-Ser phosphorylation was restored after co-incubation with Ang-(1-7), Akt-Ser phosphorylation in AngII+ Ang-(1-7)group was signifi cantly increased compared with AngII group , and not signifi cantly different from contro; yet, Ang-(1-7)exerts no effect on AngII inhibited IR-`-Tyr phosphorylation, IR-`-Try phosphorylation in AngII+Ang-(1-7)group was not signifi cantly different from AngII group, and was signifi cantly decreased compared with control. In conclusion, AngII can signifi cantly inhibit insulin-stimulated IR-`-Tyr and Akt-Ser phosphorylation in NIT-1; Ang-(1-7)also has no regulation on insulin-stimulated IR-`-Tyr and Akt-Ser phosphorylation in NIT-1; co-incubation with Ang-(1-7)can revert the inhibitory effect of AngII on Akt-Ser phosphorylation.

2767-PO Relationship Between 25-Hydroxyvitamin D and Pancreatic Islet Ƶ Cell Function in Patients With Type 2 Diabetes Mellitus JINGJING GUO, ZHENG-DA XIAO, XIN LIU, XIA XUE, YONG LU, XIAO YIN, Jinan, China, Beijing, China This study is aimed to investigate the relationship between 25-Hydroxyvi- tamin D (25-OH-D) and pancreatic islet ` cell function under different glu- cose tolerance statuses. Seventy-one patients with type 2 diabetes mellitus (DM group), 65 subjects with impaired fasting glucose/impaired glucose tol- Supported by: Japanese Ministry of Education, Science, Sports, Culture and erance (IFG/IGT group), and 69 normal control subjects (NGT group) under- Technology went 75g oral glucose tolerance test. All the subjects had not taken vitamin D and calcium in 3 months. Fasting serum 25-OH-D was assayed by ELISA. The area under the curve of the insulin secretion (IAUC), homeostasis model 2769-PO assessment for ` cell function index (Homa-`), insulin sensitivity index (ISI) Zinc Decreases Reduction-Induced Precipitation of Insulin Obesity and homeostasis model assessment for insulin resistance index (Homa-IR) KARINA SCHNITTKER, TSU-SHUEN TSAO, Tucson, AZ were calculated to explore their relationships with 25-OH-D. Levels of 25- Insulin is known to precipitate upon reduction of the disulfi de bonds that PUBLISHED ONLY OH-D, ISI and Homa-` in DM group and IFG/IGT group were signifi cantly link A and B chains. Redox states in various cellular compartments are tightly Integrated Physiology/ lower than that in NGT group (P<0.05). Homa-IR in DM group and IFG/IGT regulated to prevent oxidative stress and damage to proteins. Endoplasmic group was signifi cantly higher than that in the NGT group(P<0.05). Pearson reticulum stress, a prominent feature in pancreatic `-cells associated with correlation analysis and partial correlation analysis showed that 25-OH-D type 2 diabetes, results in a reduced redox environment. The extent to which was positively correlated with fasting insulin (FINS), Homa-`(P<0.05). Mul- insulin is reduced within storage granules is currently unknown, as are strat- tiple stepwise regression analysis showed that 25-OH-D was one of the egies employed by cells to minimize insulin reduction and precipitation. We infl uential factors of pancreatic islet ` cell function in patients with type show in the present study that signifi cant amounts of insulin can be reduced 2 diabetes mellitus. So we concluded level of serum 25-OH-D was low in in vitro at the redox state previously reported for secretory pathway. As patients with type 2 diabetes mellitus. 25-OH-D was closely related with the zinc is found at high levels within insulin storage granules and is both coor- function of the pancreatic islet ` cell and was one of the infl uential factors dinated and bound by insulin hexamers, we investigated the effect of zinc of pancreatic islet ` cell function. on precipitation of insulin upon reduction. Zinc specifi cally decreased the

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A689 INTEGRATEDCATEGORY PHYSIOLOGY—LIVER

precipitation of insulin in vitro upon treatment with DTT or the non-thiol- and JDRF are cosponsoring these studies. Results from the fi rst study in 20 containing reducing agent TCEP. Other divalent cations including copper, healthy, obese (BMI 27-38) men and women will be available in 2012 and magnesium, calcium, and manganese were unable to prevent insulin precipi- planned for submission as late-breaking ADA abstracts this year. tation. While zinc slowed the reduction of insulin as determined by tricine Supported by: FNIH Biomarkers Consortium SDS-PAGE, it did not prevent the complete reduction of insulin, suggesting the effect of zinc on insulin precipitation is independent of insulin reduction. 2772-PO In addition, zinc had no effect on reduction of DTNB. Our results suggest that White Blood Cell Count is Independently Associated With Insulin zinc may be able to mitigate potential reduction and precipitation of insulin Resistance in Type 1 Diabetic Patients within storage granules, thus presenting a role for zinc that was previously TOMISLAV BULUM, LEA DUVNJAK, Zagreb, Croatia unknown. Further studies must be completed to determine the biochemical Previous studies have reported signifi cant independent associations of basis by which zinc prevents insulin precipitation and to determine in vivo routinely measured hematological parameters with insulin resistance and relevance. type 2 diabetes. To investigated the relationship between red blood cell count (RBC), hemoglobin (Hgb), white blood cell count (WBC), platelets and 2770-PO insulin resistance in type 1 diabetes we divided 353 patients according to median estimated glucose disposal rate (eGDR=9.72 mg/kg-1min-1) into lower and higher-insulin sensitivity group. None showed signs of acute or chronic WITHDRAWN infl ammatory, renal and cardiovascular diseases. Estimated glucose disposal rate was calculated using the equation: eGDR = 24.31 - (12.22xwaist-to-hip ratio (WHR)) - (3.29xhypertension) - (0.57xHbA1c). Patients with lower insulin sensitivity in comparison to patients with higher insulin sensitivity showed signifi cantly higher WBC and Hgb (7.1 vs 6.3x109/L; 143 vs 136 g/L, p=0.001). RBC, Hgb and WBC were signifi cantly correlated with eGDR (r=-0.12, -0.21 and -0.14, respectively, p)0.01), and WHR (r=0.38, 0.44, and 0.16, respec- tively, p)0.001). In a multiple logistic regression analysis after adjustment for covariates, WBC was identifi ed as an independent predictor of progres- sion to insulin resistance (OR=1.28, 95% CI=1.13-1.45, p<0.001). In patients in the 4th quartile of WBC, in comparison to patients in the 1st quartile of WBC, the risk of progression to insulin resistance increased by a factor of 4.41 (Figure 1). The signifi cant independent association of WBC with the pro- gression to insulin resistance suggests a role of subclinical infl ammation in its pathogenesis.

2771-PO Characterization of Measures of Beta Cell Function: A Precompeti- tive Collaboration of the Foundation for the NIH Biomarkers Consor- tium (FNIH BC) DAVID A. FRYBURG, MARIA T. VASSILEVA, Bethesda, MD INTEGRATED PHYSIOLOGY—LIVER Reliably measuring beta cell (bc) function in multicenter trials is critical for conduct of long-term studies of bc failure. Of deployable methods, little is 2773-PO known about test-retest variability of meal tolerance test (MTT) or the maxi- Estrogen Improves Diet-Induced Impaired Glucose Intolerance mal stimulation test (MST) with arginine or glucagon. Although MTT has by Inhibiting Gluconeogenesis and Lipogenesis Independent of been used in many studies, meal composition and caloric load have varied Insulin considerably. Moreover, there are no data using the same methods across PAN-WEI MU, PING JIANG, WEI JIANG, ZHUO-ZHUO REN, ZHI TAN, SHU-HUI spectrum from health to prediabetes to type 2 diabetes (T2DM), nor results ZHENG, KE-WEN ZHOU, YA-XING ZHANG, TING-HUAI WANG, Guangzhou, China using newer modeling techniques. Methodologic studies of reproducibility of Estrogen has been reported to improve glucose metabolism by enhance- metabolic tests are expensive. As the results benefi t the metabolic research ment of insulin secretion and improvement of insulin sensitive. However, it

Obesity community, sharing responsibility for design and execution among the stake- is still unclear whether it can ameliorate blood glucose through other mecha- holders, including academia, non-profi ts (ADA and JDRF), government (NIH nisms. In present study, diabetic rats were induced by high caloric diet. 17be- and FDA) and the pharmaceutical industry, is best. To that end, a neutral en- ta-estradiol (E2) was injected daily with a dosage of 20μg-1kg-1.Fasting blood PUBLISHED ONLY Integrated Physiology/ vironment is necessary to bring together expertise and funding and ensure glucose and insulin were examined prior to and after 4-month E2 treatment. delivery of results to the scientifi c community. The FNIH BC was created for Insulin sensitive was calculated. Gluconeogenesis and lipogenesis in liver this general purpose in 2006. In 2011, a team of experts from all stakeholder were also detected. The results showed E2 could low fasting blood glucose groups, the Beta Cell Project Team of the FNIH BC, designed a series of fun- signifi cantly but had no effect on insulin secretion or sensitive. It could in- damental clinical studies to facilitate use of standardized methodologies hibit phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase in multi-center trials. A 3-year plan of studies was designed to: 1. Deter- (G6pase), and Forkhead box protein O1 (FoxO1). Furthermore E2 suppressed mine within- and between-subject variability of MTT and MST responses in the activities of sterol regulatory element-binding protein (SREBP) and car- healthy,prediabetic (IFG/IGT) and T2DM subjects; compare gender responses bohydrate responsive element-binding protein (ChREBP). Taken together, the 2. Compare beta cell function as estimated by MTT and FSIVGT 3. Determine results suggest estrogen could improve glucose metabolism by inhibition impact of testing sequence 4. Defi ne effect of medications This presentation of gluconeogenesis and lipogenesis in liver, which is independent of insulin will describe the plan. Nine pharmaceutical companies, the NIH, FDA, ADA, signaling pathway.

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A690 INTEGRATEDCATEGORY PHYSIOLOGY—LIVER

Signal intensities were used for determining fat content. Then the fat total signal area (FTSA) ratio was calculated. For statistical analysis, 2x2 cross- over ANOVA and Pearson’s correlations were used. FTSA was reduced in response to both regimens (p<0.001), more in B (-3.4; 95% CI, -3.8 to -3.1 % in A vs. -4.2; 95% CI, -4.5 to -3.8% in B; p=0.03). Body-mass-index (BMI) was reduced in both regimens (p<0.001), more in B (-0.82; 95% CI -0.94 to -0.69 kg.m-2 in A vs. -1.23; 95% CI -1.4 to -1.17 kg.m-2 in B; p<0.001). Fasting gly- cemia decreased in both regimens (p<0.001), more in B (-0.47; 95% CI -0.57 to -0.36 mmol/l in A vs. -0.78; 95% CI -0.89 to -0.68 mmol/l in B; p=0.004). Decrease in FTSA correlated strongly positively with decrease in fasting glycemia (r=+0.56; p<0.001). After adjustment for changes in BMI this as- sociation remained signifi cant (r=+0.28; p=0.05). Prolonged fasting reduced hepatic fat content more than a diet with the same caloric restriction divided in more frequent meals in patients with T2D. This reduction was associated with a decrease in fasting glycemia, independently on changes in BMI. Supported by: Grant VZ MZO 00023001

2776-PO Serum Fetuin-A Concentrations in Subjects With Prediabetes and Nonalcoholic Fatty Liver Disease HORNG-YIH OU, YIH-CHING YANG, HUNG-TSUNG WU, JIN-SHANG WU, FENG- HWA LU, CHIH-JEN CHANG, Tainan, Taiwan Background: Heptatic steatosis and prediabetes are associated with an increased cardiovascular risk. Fetuin-A, a liver-derived glycoprotein that impairs insulin signaling, is independently associated with risk of cardio- 2774-PO vascular diseases in previous studies. Although serum fetuin-A levels are Absence of Intestinal Gluconeogenesis in Type 1 Diabetic Rats: In associated with liver fat and metabolic syndrome, the association among Vitro and In Vivo Evidence fetuin-A, prediabetes, and nonalcoholic fatty liver disease (NAFLD) is not GABRIEL BAVEREL, GÉRARD MOINET, GUY MARTIN, BERNARD FERRIER, AGNÈS known. Aims: To investigate the relationship between serum fetuin-A levels CONJARD-DUPLANY, Lyon, France and prediabetes in subjects with or without NAFLD. Methods: A total of Recent studies have reported that the small intestine of diabetic rats sub- 510 age- and sex-matched subjects with normal glucose tolerance (NGT), stantially contributes to systemic glucose production via gluconeogenesis. impaired fasting glucose (IFG), and impaired glucose tolerance (IGT) with or Since this would (i) represent a major change in the classical view that only without NAFLD were recruited. NAFLD was assessed by abdominal ultra- the liver and the kidneys have a gluconeogenic capacity and (ii) have major sound. Serum fetuin-A concentrations were compared between groups. The consequences in the pathophysiology of diabetes, we have re-examined this association with clinical and metabolic parameters was examined. Results: question. Metabolically viable intestinal cells from adult fed and 36h-fasted The presence of NAFLD signifi cantly increases serum fetuin-A levels in sub- streptozotocin-diabetic Sprague Dawley rats were incubated in the pres- 13 jects with NGT and prediabetes. As compared with NGT, IGT, but not IFG, ence of 5 mM [3- C]glutamine in the absence and the presence of 5 mM signifi cantly increases serum fetuin-A levels in subjects with or without NA- unlabeled glucose. Substrate removal and product accumulation were mea- 13 FLD. Serum fetuin-A concentrations were positively related to post-load 2h sured by enzymatic and C-NMR spectrometry. Arterio-portal concentration glucose, BMI, triglyceride, homeostasis model assessment (HOMA-IR), but difference measurements were also performed in 36h-fasted diabetic rats. 13 negatively associated with age, HDL-cholesterol, and adiponectin. In mul- Despite the removal of C-glutamine and the formation of glutamate, lac- tiple regression analysis, age, “IGT vs. NGT”, and “IGT+NAFLD vs. NGT” were tate, alanine, citrulline and 13CO at high rates, glucose synthesis by isolated 2 independently associated factors of fetuin-A levels after adjustment for car- small intestinal cells from both fed and fasted diabetic animals was not diovascular risk factors and adiponectin. Conclusions: Both NAFLD and IGT observed and this was not due to the re-utilization of glucose synthesized were independently associated with serum fetuin-A concentrations after from glutamine. Arterio-portal concentration difference measurements for adjusting for cardiometabolic risk factors. This aspect of elevated fetuin-A glucose, lactate, pyruvate, alanine, glycerol, glutamine, glutamate in 36h- levels could have a clinical implication in the increased cardiovascular risk fasted diabetic rats showed that the intestine neither removed nor added associated with NAFLD and IGT. glucose in net amounts. Lactate and alanine were released in net amounts Supported by: NSC-95-2314-B-006-048 by the intestine. Only 15% of circulating glutamine, which has been reported to be the main if not the exclusive intestinal gluconeogenic precursor in dia- betic rats in vivo, was found to be taken up. Assuming that the glutamine 2777-PO taken up was completely converted into glucose, a very unlikely hypothesis, Hepatic Chemerin is Induced in Non-Alcoholic Fatty Liver Disease this would have diluted the circulating glucose by less than 1%, a percent- CHRISTA BUECHLER, JOSEF WANNINGER, SABRINA BAUER, KRISTINA EISING- age which is not measurable in a reliable manner by currently available ana- ER, DORIS SCHACHERER, CLAUS HELLERBRAND, THOMAS S. WEISS, ANDREAS lytical methods. The present results indicate that the existence of intestinal SCHAFFLER, Regensburg, Germany gluconeogenesis remains to be demonstrated in diabetic animals. Chemerin is a recently described adipokine whose adipose tissue and serum levels are increased in obesity. Chemerin is also expressed in the liver, and here, levels of chemerin in liver cells and non-alcoholic fatty liver 2775-PO disease (NAFLD) were studied. Primary human liver cells and liver tissue Prolonged Fasting Reduces Hepatic Fat Content in Patients With of patients with and without liver steatosis were used. Human liver tissue Type 2 Diabetes was obtained according to the guidelines of the charitable state-controlled Obesity HANA KAHLEOVA, LENKA BELINOVA, MILAN HAJEK, MONIKA DEZORTOVA, foundation Human Tissue & Cell Research with the patient’s informed con- MARTIN HILL, TEREZIE PELIKANOVA, Prague, Czech Republic sent. To induce non-alcoholic steatohepatis (NASH), mice were fed a Paigen Prolonged Fasting Reduces Hepatic Fat Content in Patients with Type 2 PUBLISHED ONLY diet for 12 weeks or a methionine-choline defi cient diet (MCD) for 6 weeks. Integrated Physiology/ Diabetes Increased hepatic fat content is associated with type 2 diabetes Chemerin mRNA has been detected in primary human hepatocytes (PHH), (T2D). Caloric restriction is crucial in its treatment typically apportioned into epithelial cells and hepatic stellate cells (HSC) but not in Kupffer cells and fi ve or six small meals during the day. The aim of our study was to compare bile-duct cells. Chemerin in the supernatants of PHH is signifi cantly higher the effect of six meals vs. two meals a day with the same caloric restric- compared to HSC and human adipocytes. In patients with fatty liver hepatic tion on hepatic fat content in subjects with T2D. In a randomized, crossover and serum chemerin is similarly high as in probands with non-steatotic liver. study, we assigned 54 patients with T2D to follow two regimens of a hy- In rodent NASH liver chemerin is increased. Feeding Paigen diet elevates pocaloric diet (-500 kcal/day), each for 12 weeks: six meals a day (A), and body weight and serum chemerin. Despite weight loss MCD fed mice have two meals a day (B). The diet in both regimens had the same macronutrient similar serum chemerin as the respective control animals. Chemerin in con- and energy content. We measured the hepatic fat content by the proton centrations found in serum in obesity impairs insulin mediated Akt phospho- magnetic resonance spectroscopy performed by 3T MR scanner (Magnetom rylation and suppression of glucose release in PHH. The antidiabetic drug - Trio Siemens). Spectra were obtained from three different parts of the liver.

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A691 INTEGRATEDCATEGORY PHYSIOLOGY—LIVER

metformin reduces cellular and soluble chemerin in PHH as has already been 2780-PO described in adipose tissue. In conclusion current data show that PHH are the major source of hepatic chemerin and increased synthesis in NASH may contribute to higher systemic levels and insulin resistance. Supported by: German Research Foundation WITHDRAWN 2778-PO O-GlcNAc Cycling: Association With Genetic Susceptibility to Fructose for Hepatic Lipogenesis and Metabolic Disorders OSAMU SEKINE, YOSHIHIKO NISHIO, MASATERU USHIO, KEIKO KOSAKA, KAT- SUTARO MORINO, TAKESHI YOSHIZAKI, SATORU UGI, ATSUNORI KASHIWAGI, HIROSHI MAEGAWA, Otsu, Shiga, Japan, Kagoshima, Japan Inappropriate nutrient balances cause metabolic derangements. A dy- namic cycle of O-linked N-acetylglucosamine (O-GlcNAc) addition and re- moval is catalyzed by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), respectively, in a process which serves as the fi nal step in a nutrient-driven ‘hexosamine-signaling pathway’. Evidence points to a role for O-GlcNAc cy- cling in diabetes and insulin resistance. We evaluated the relationship be- tween O-GlcNAc cycling and metabolic disorder in the mouse hepatocytes with high fructose diet. Four-week-old male mice of CBA/JN and DBA/2N strains were fed in the control or high fructose diet for 4 weeks, and then liver was extracted. We investigated expression levels of O-GlcNAc modi- fi ed protein in the nucleus using Western blot analysis and mRNA levels of O-GlcNAc cycling-related genes using real-time PCR. Compared with control 2781-PO diet, fructose diet elevated the levels of serum insulin and intrahepatic trig- lyceride content by 2-fold (p < 0.05), respectively, and increased the levels of SREBP-1c mRNA expression by 2.7-fold (p < 0.05) in the CBA/JN mice, but WITHDRAWN not in the DBA/2N mice. Fructose diet increased the expression patterns of O-GlcNAc modifi ed protein and also increased the levels of OGT mRNA expression by 1.3-fold (p < 0.05) in the CBA/JN mice, but not in the DBA/2N mice, compared with control diet. Fructose diet decreased the levels of OGA mRNA expression in the both strains. Similar results were observed 2782-PO in primary hepatocytes after exposure to fructose. The mRNA expression of Akt2 is Required for Insulin-Dependent Hepatic Glycogen Accumu- GFAT1, a regulator for hexosamine-signaling pathway, was less by 0.6-fold (p lation < 0.05) in the DBA/2N mice than in the CBA/JN mice. These results suggest MIN WAN, MORRIS J. BIRNBAUM, Philadelphia, PA that the regulation of hexosamine-signaling pathway and O-GlcNAc cycling Previously researches from our laboratory and others have shown that refl ects on the genetic susceptibility to hepatic lipogenesis and metabolic Akt2, the major Akt isoform in the liver, is required for high-carbohydrate disorders induced by high fructose diet. diet-induced de novo lipogenesis and high-fat diet-induced hepatic steato- sis; however, the glycemic profi le is only minimally affected. Here we re- 2779-PO port that deletion of Akt2 from liver led to modest insulin resistance with a blunted suppression of hepatic glucose production by insulin during an euglycemic-hyperinsulinemic CLAMP. Studies in perfused mouse liver indi- cated that the insulin failed to inhibit hepatic mobilization in livers without WITHDRAWN Akt2, while gluconeogenesis was not altered. In vivo, liver-specifi c dele- tion of Akt2 resulted in defective hepatic glycogen accumulation after an euglycemic-hyperinsulinemia CLAMP or 4-hour refeeding. Interestingly, the canonical Akt-GSK3_/`-GS pathway, which has been suggested to mediate the insulin effects on glycogen metabolism, was not disrupted by deletion of Akt2. These data showed that Akt2 inhibits hepatic glucose production by modulating glycogen metabolism in the liver, and the modulation is indepen- dent of the canonical Akt-GSK3_/`-GS pathway. Supported by: NIH (RO1 DK56886, PO1 DK49210)

2783-PO Hepatic and Skeletal Muscle Lipid Content in Healthy Male Adoles- cents from a Rural Region in South India NIHAL J. THOMAS, ROSHAN LIVINGSTONE, LOUISE G. GRUNNET, MEREDITH HAWKINS, ALLAN VAAG, BJØRN QUISTORFF, IB C. BYGBJERG, Vellore, India, Copenhagen, Denmark, New York, NY Asian Indians have a high prevalence of Type 2 diabetes mellitus (T2DM), Obesity and low birth weight (LBW) subjects have an increased risk of developing T2DM. Whether that is mediated via ectopic fat disposition in liver and mus- cle is unknown. We assessed the association of ectopic lipid with insulin PUBLISHED ONLY Integrated Physiology/ sensitivity for LBW and normal birth weight (NBW) subjects from a rural region in South India. The study included 117 healthy non-diabetic male ado- lescents aged 18 -22yrs, of which 61 were of LBW (BW <2450g) and 56 NBW; (BW 3100> and <3500 g). The mean BMI of LBW and NBW were 19.0kg/m2 and 19.5kg/m2 respectively. All underwent a hyperinsulinemic-euglycemic clamp for evaluating insulin sensitivity. Hepatic cellular lipid (HCL) and intramyocellular lipid (IMCL) from soleus muscle was quantifi ed from 1H magnetic resonance spectroscopy (MRS) using a 3T MR scanner. The HCL content was quantifi ed with water as an internal reference, and IMCL in the soleus muscle was assessed as IMCL/ total creatine (tCr). Mean HCL con- tent for LBW and NBW subjects were 0.43% and 0.31% respectively and in

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A692 INTEGRATEDCATEGORY PHYSIOLOGY—LIVER most cases, lipid peaks were almost undetectable. Mean IMCL/tCr content 2786-PO for LBW and NBW were 6 and 5.92 respectively. Signifi cant negative cor- Androgen Deprivation Worsens High-Fat Diet-Induced Glucose In- relations were observed between HCL and birth weights (LBW: r = -0.34, p = tolerance and Hepatic Fatty Degeneration 0.015; NBW: r = -0.34, p = 0.018). No signifi cant correlations were observed TAKAFUMI SENMARU, MICHIAKI FUKUI, HIROSHI OKADA, YUSUKE MINEOKA, between IMCL/tCr and birth weights. No signifi cant correlation between HIROYA IWASE, MAI ASANO, MASAHIRO YAMAZAKI, HIROSHI OBAYASHI, GOJI insulin sensitivity (M-value) and HCL (LBW: r=-0.04, p=0.78; NBW: r=-0.08, HASEGAWA, NAOTO NAKAMURA, Kyoto, Japan p=0.58), IMCL/tCr(LBW: r=0.24 p=0.10 NBW: r=-0.25 p=0.08) were observed Androgen decline causes insulin resistance. However, the underlying mech- in both groups. In the LBW, HCL and Insulin levels showed a signifi cant cor- anisms remain unclear. The aim of this study was to investigate the effects of relation (r=0.669, p=0.00) prior to the clamp. This fi nding was not replicated androgen deprivation on glucose and lipid metabolism in liver. Male C57BL/6J in the NBW group. Low HCL in both LBW and NBW subjects could be due to mice were either castrated (CAS) or sham-operated (SHAM) at 7 wk of age. age and low BMI. In this population, it may be premature to conclude that Starting at 8 wk of age, then, mice were fed high-fat diet (HFD) or standard diet the early onset of T2DM for LBW maybe due to ectopic fat deposits in the (SD) for 3 wk. Body weight, fasting blood glucose levels, serum insulin levels, liver. A reassessment in the next decade may be warranted. serum triglyceride levels and hepatic lipid accumulation signifi cantly increased Supported by: Copenhagen University and Biocampus in HFD-fed mice compared with those in SD-fed mice. Fasting blood glucose levels and hepatic lipid accumulation increased more in CAS/HFD than those 2784-PO in SHAM/HFD. In intraperitoneal glucose tolerance test, HFD-feeding deterio- rated glucose tolerance. Castration did not affect glucose tolerance in SD-fed mice, however, blood glucose levels at 30, 60 and 120 min after glucose load WITHDRAWN were signifi cantly higher in CAS/HFD than those in SHAM/HFD. Hepatic gene analyses revealed that the expression levels of sterol regulatory element bind- ing protein-1c (SREBP-1c) were higher in HFD-fed mice than those in SD-fed mice. Furthermore, the expression levels of SREBP-1c were signifi cantly higher in CAS/HFD than those in SHAM/HFD. Similarly, the expression levels of fatty acid synthase, a downstream target of SREBP-1c in hepatic insulin signaling pathway, were signifi cantly higher in CAS/HFD than those in the others. In- versely, the expression levels of insulin receptor substrate-2 were signifi cantly lower in CAS/HFD than those in the others. In histological analysis, hepatic fatty degeneration was found in HFD-fed mice and it was more evident in CAS/HFD than in SHAM/HFD. Collectively, these results indicate that andro- gen deprivation worsens HFD-induced glucose intolerance and hepatic fatty degeneration due to the enhancement of hepatic insulin resistance.

2787-PO

WITHDRAWN

2785-PO Time Sequence of the Intensifi cation of the Liver Glucose Produc- tion Induced by High-Fat Diet in Mice HELENTON C. BARRENA, SIMONI OBICI, THAUANY M. TAVONI, RUI CURI, RO- BERTO B. BAZOTTE, Maringá, Brazil, São Paulo, Brazil It is well established that the development of insulin resistance show a temporal sequence in different organs and tissues. Moreover, considering that the main aspect of insulin resistance in liver is a process of glucose overproduction from gluconeogenesis we investigated if this metabolic change also shows temporal sequence. For this purpose, a well established experimental model of insulin resistance induced by high-fat diet (HFD) was used. The mice received HFD (HFD group) or standard diet (COG group) from 1, 7, 14 or 56 days. HFD group showed increased (P < 0.05 vs. COG) epididymal, retroperitoneal and inguinal fat weight from day 1 until day 56.

In agreement with these results HFD group also showed higher body weight Obesity (P < 0.05 vs. COG) from day 7 until day 56. Moreover, the changes induced by HFD on liver gluconeogenesis were progressive, since the increment (P < PUBLISHED ONLY 0.05 vs. COG) in glucose production from L-lactate, glycerol, L-alanine and L- Integrated Physiology/ glutamine occurred 7, 14, 56 and 56 after the introduction of the HFD sched- ule, respectively. Furthermore, glycemia and cholesterolemia increased (P < 0.05 vs. COG) 14 days after starting the HFD schedule. Taken together the results suggest that the intensifi cation of liver gluconeogenesis induced by a HFD is not a synchronous “all or nothing process” but is specifi c for each gluconeogenic substrate and is integrated in a temporal manner with the progressive augmentation of fasting glycemia. Supported by: CNPq (Grants 563870/2010-9), FAPESP and Araucaria Foundation

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A693 INTEGRATED PHYSIOLOGY—MACRONUTRIENTCATEGORY METABOLISM AND FOOD INTAKE

2788-PO 2791-PO Gamma Glutamyl Transferase (GGT) Levels Refl ect Insulin Resis- tance in Subjects With Persistent Impaired Glucose Tolerance RAM JAGANNATHAN, SNEHALATHA CHAMUKUTTAN, NANDITHA ARUN, SAMITH SHETTY, DESMOND G. JOHNSTON, RAMACHANDRAN AMBADY, Chen- WITHDRAWN nai, India, London, United Kingdom Asian Indians have lower body mass index (BMI), higher waist circum- ference and high body fat percentage and more insulin resistance (IR) than many other races. Impaired Glucose Tolerance (IGT) is associated with higher insulin resistance. This analysis was done to evaluate the associa- tion of Gamma Glutamyl Transferase (GGT) with IR, `-cell function and other cardiometabolic risk factors in Asian Indian subjects with persistent IGT. In a cohort of 275 men with persistent IGT (mean age 46.1 + 4.7 years), baseline values of plasma glucose and insulin were measured at 0, 30 and 120 min- utes of OGTT, insulin resistance (HOMA-IR) and `-cell function (incremental insulin/30 glucose,6 I/G) were calculated, BMI, waist circumference, body fat percentage (bioimpedence method) were measured, and fasting serum cholesterol, triglycerides(TG), HDL-cholesterol (HDL-C) were estimated. Correlations of GGT with above parameters were assessed. Median value of GGT was 23.2 IU/l in IGT subjects. GGT showed no correlation with the anthropometric variables. It showed signifi cant correlation with HOMA-IR (r=0.209, p<0.0001), TG (r=0.555,p<0.0001), fasting insulin (r=0.202,p<0.001) HDL-C (r=-0.138,p=0.023) and with cholesterol (r=0.196,p<0.001) (all values log transformed). GGT showed no correlation with the `-cell function. Unlike in many other populations, GGT levels were abnormal even without obesity in Asian Indians with IGT. GGT levels were signifi cantly associated with IR, independent of the anthropometric variables and it showed association with lipid parameters and fasting insulin, known to be surrogate markers of IR. GGT is a simple surrogate measurement of IR in Asian Indian men with IGT and may be a predictor of future diabetes. Supported by: UK India Education and Research INTEGRATED PHYSIOLOGY—MACRONUTRIENT 2789-PO METABOLISM AND FOOD INTAKE 2792-PO WITHDRAWN Ingestion of Leucine and Glycine With Glucose Produces a Partially Additive Effect on Plasma Glucose Despite a Less than Additive Ef- fect on Serum Insulin JENNIFER F. IVERSON, MARY C. GANNON, FRANK Q. NUTTALL, Minneapolis, MN Our laboratory previously demonstrated that the majority of individual 2790-PO amino acids stimulate insulin and attenuate the plasma glucose response Impact of Fatty Liver on Insulin Resistance in Japanese Patients when ingested with glucose. The objective of this study was to determine With Type 2 Diabetes whether ingestion of two amino acids simultaneously with glucose would HIROSHI TAKATA, MARI INOUE, AYAKO ISHIBASHI, YOSHITAKA KUMON, YOSH- result in a synergistic, additive or less than additive effect on glucose and IO TERADA, SHIMPEI FUJIMOTO, Kochi, Japan insulin areas compared to ingestion of the amino acids individually. Leucine Fatty liver links metabolic disorders including obesity, diabetes, dyslipi- (Leu) and glycine (Gly) were chosen because they were two of the most po- demia, and hyperinsulinemia. However, the impact of fatty liver on insulin tent amino acids in lowering plasma glucose when given individually. Nine resistance in patients with type 2 diabetes (T2DM) is not fully understood. healthy subjects were studied on 4 separate days. Test meals were given Contributing factors predicting insulin resistance were analyzed in T2DM at 0800. The fi rst meal was a water control. Subjects then received 25 g patients with or without fatty liver. We recruited 114 Japanese patients [76 glucose or Leu + Gly (1 mmol/kg lean body mass each) with or without 25 g men and 38 women, age: 63±12 years, BMI: 25.2 ±4.4 (mean±SD)] treated glucose in random order. For 2.5 hours after ingestion of the test meals blood without insulin. Total serum concentration of adiponectin, a major adipokine glucose and insulin levels were measured frequently and net areas under the that ameliorates insulin resistance, was measured using an enzyme-linked curve calculated. The mean fasting value was used as baseline. The results immunosorbent assay (Otsuka, Japan). Fatty liver was examined by ultra- were compared with data obtained previously when glucose was ingested sonography. QUICKI, an index of insulin sensitivity, was calculated using with each individual amino acid. Ingestion of glucose + Leu + Gly decreased fasting plasma glucose and serum IRI levels. In simple regression analysis, the glucose area response by 77% (p=0.015) and increased the insulin area QUICKI was correlated with BMI in both the fatty liver group (FG)(n=61, r=- response by 24% (p=0.046) when compared to ingestion of glucose alone. 0.56, p<0.01) and the non-fatty liver group (NFG)(n=53, r=-0.46, p<0.01), and The decrease in glucose response with Leu + Gly + glucose was partially was correlated with adiponectin in NFG (r= 0.47, p<0.01) but not in FG. Step- additive when compared to ingestion of the amino acids individually with wise multiple regression analyses showed that BMI, adiponectin, and age glucose (Leu = 50%, Gly = 58%, Leu + Gly = 77%). In contrast, the increase in

Obesity were independent contributors to QUICKI in all patients. In NFG, adiponectin insulin response seen with Leu + Gly + glucose was less than additive when (`=0.43, F=12.5) and BMI (`=-0.41, F=12.2) were independent contributors to compared to the individual amino acids ingested with glucose (Leu = 66%, QUICKI, accounting for 37.5% of the variability. In FG, BMI (`=-0.68, F=35.1), Gly = 1%, Leu + Gly = 24%). Thus, ingestion of Leu + Gly with glucose produc- PUBLISHED ONLY Integrated Physiology/ adiopnectin (`=0.29, F=6.6), HbA1c (`=-0.17, F=2.4) and age (`=-0.18, F=2.1) es a partially additive effect on glucose area response in healthy subjects were independent contributors to QUICKI, accounting for 40.0% of the vari- despite a less than additive effect on insulin area response. The mechanism ability. Taken together, these fi ndings indicate that in T2DM patients, the by which this occurs is unknown. contribution of adiponectin to insulin resistance is less in those with fatty Supported by: Department of Veterans Affairs liver compared to that in those without fatty liver, suggesting a metabolic effect of fatty liver on insulin resistance in T2DM.

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A694 INTEGRATED PHYSIOLOGY—MACRONUTRIENTCATEGORY METABOLISM AND FOOD INTAKE

2793-PO 2796-PO Role of Thyroid Transcription Factor 1 in Streptozotocin-induced Diabetic Hyperphagia JAE GEUN KIM, BYONG SEO PARK, IL SEONG NAMGOONG, YOUNG IL KIM, WITHDRAWN BYUNG JU LEE, New Haven, CT, Ulsan, Republic of Korea Thyroid transcription factor-1 (TTF-1), a homeodomain-containing tran- scription factor, is postnatally expressed in discrete areas of the hypo- thalamus and closely involved in neuroendocrine functions. We previously reported that TTF-1 transcriptionally regulates _-melanocyte-stimulating 2797-PO hormone (_-MSH) and agouti-related peptide (AgRP) in the rat arcuate nu- Moderate Amounts of Sugar Sweetened Beverages Impair Hepatic cleus and affects feeding behavior via the melanocortin pathway. Moreover, Insulin Sensitivity and Lipid Metabolism, Including Beta-Oxidation hypothalamic TTF-1 expression was signifi cantly increased in the fasting of Fatty Acids, in Healthy Young Men—A Randomized Controlled Trial induced-hypoleptinemia and hypoinsulinemia condition. This study aims to MICHEL HOCHULI, ISABELLE AEBERLI, PHILIPP GERBER, HEINZ TROXLER, LISA identify TTF-1 function in the hyperphagia induced by type 1 diabetes. We SZE, STEFANIE MURER, GIATGEN SPINAS, KASPAR BERNEIS, Zurich, Switzerland fi rst observed expression of TTF-1 in the hypothalamus of streptozotocin High quantities of sugar-sweetened beverages (SSB) have unfavourable (STZ)-induced diabetic rats. These animals revealed signifi cantly increased effects on glucose and lipid metabolism in obese subjects, but the effects level of hypothalamic TTF-1 expression, and severe weight loss and strong of moderate amounts in normal weight subjects are highly debated. The aim appetite as compared with control animals. Surprisingly, the STZ-induced of the present study was to investigate the effects of moderate amounts of hyperphagia was almost completely disappeared by knocking-down hypo- various sugars in SSB on glucose and lipid metabolism in healthy young men. thalamic TTF-1 synthesis with intracerebroventricular (icv) administration of Nine subjects were studied in a controlled cross-over trial. Four three-week an antisense (AS) oligodeoxynucleotide (ODN). Moreover, increased AgRP interventions were assigned in random order: Medium fructose (MF, 40 g/ and decreased _-MSH expression, exerted by the STZ-induced hyperphagic day); high fructose (HF, 80 g/day), sucrose (HS, 80 g/day), glucose (HG, 80g/ condition, were respectively restored to the normal control level by the AS day), all provided as 3x 200ml sweetened beverages per day. Euglycemic ODN. Icv administered leptin attenuated effect of STZ-induced hyperphagia hyperinsulinemic clamps were performed at the end of each intervention on the hypothalamic TTF-1 and neuropeptide expression. These results sug- 2 with [6,6]- H2-glucose labeling to measure endogenous glucose production. gest a strong appetite control action of TTF-1 in the type I diabetes. Acylcarnitine profi les (fasting and at the end of each clamp) were measured by mass spectroscopy. Hepatic glucose production during the clamp was 2794-PO signifi cantly higher after HF compared to HG (ratio clamp/baseline 0.41 ± Diet Oleic Acid (OA) Improves Metabolic Risk Factors in Young 0.11 vs. 0.30 ± 0.10, p=0.009), indicating reduced hepatic insulin sensitivity Adults: Lipidomic Analysis of Sexually Dimorphic Responsiveness after the HF diet. Long-chain fasting acylcarnitines were increased after HF C.L. KIEN, JANICE Y. BUNN, MATTHEW E. POYNTER, ROBERT STEVENS, JAMES and HS diets, but not after HG (e.g. C16 palmitoylcarnitine HF 1.36, HS 1.34 vs BAIN, OLGA IKAYEVA, NAOMI K. FUKAGAWA, CATHERINE M. CHAMPAGNE, HG 0.96 μmol/l, p < 0.05). Depending on the diet, the clamp had differential KAREN CRAIN, TIMOTHY R. KOVES, DEBORAH M. MUOIO, Colchester, VT, Bur- effects on the levels of long chain (intermediates prior to beta-oxidation) ver- lington, VT, Durham, NC, Baton Rouge, LA sus short and medium chain acylcarnitines. Overall, the present study shows Mediterranean diets rich in OA are associated with reduced risk of coro- adverse effects already of moderate amounts of fructose on hepatic insulin nary artery disease and type 2 diabetes relative to Western diets enriched sensitivity. In addition it is the fi rst study in humans analysing acylcarnitine in palmitic acid (PA). To gain insight into mechanisms underlying these ob- profi les after ingestion of moderate amounts of various sugars, and suggests servations, we applied comprehensive lipidomic profi ling tools in 18 healthy differences in the regulation of fatty acid beta-oxidation after diets contain- adults (9 women) enrolled in a randomized, cross-over trial comparing a high ing fructose, i.e. limited substrate clearance and/or enhanced substrate fl ow PA diet (HPA) to a low PA/high OA diet (HOA). Principal components analysis towards this pathway. (PCA)(329 variables) identifi ed one major factor (F1) that was affected by diet (P < 0.0001) and strongly weighted by the PA/OA ratio of serum and muscle 2798-PO lipids. HOA lowered the serum LDL/HDL ratio; adjusting for F1 abrogated this A Novel Technique to Investigate Post-Prandial Responses to Sugar relationship. In women only, during the HOA diet, insulin sensitivity (SI) and Solutions in Rats pancreatic beta cell responsiveness (DI), assessed by intravenous glucose SYED SUFYAN HUSSAIN, ERROL RICHARDSON, NIKI BUCKLEY, KYLIE E. BEALE, tolerance testing, were respectively 63% (P = 0.04) and 46% (P = 0.02) higher, STEPHEN R. BLOOM, JAMES V. GARDINER, London, United Kingdom and F1 inversely correlated with both SI (Spearman r = - 0.786, P = 0.02) and Administration of oral substances, such as glucose, to conscious rodents DI (r = - 0.750, P = 0.05); moreover, the increase in SI was positively modifi ed forms an essential part of post-prandial investigations. This is normally by pre-study physical fi tness (r = 0.90, P = 0.001). In women: (1) Change in SI achieved by oral gavage, which bypasses cephalic responses and may also tended to inversely correlate with change in serum total ceramide concen- induce stress. We present a novel technique to administer sweet solutions to tration (r = -0.667, P = 0.07), which was higher during HPA (fasting and fed rats, which relies on voluntary consumption. This is likely to maintain cephal- states P 0.01). (2) During the fed state on the HPA diet, the sum of medium ic responses and avoids the need for excessive handling during administra- chain (MC) acylcarnitines (AC)(MCAC) and the MCAC/long chain AC ratio in tion. The utility of this technique in delivering 2.5g/kg glucose in male Wistar blood inversely correlated with SI (respectively r = - 0.783, P = 0.01 and r = - rats was assessed following an overnight fast. After 5 days of training, 95% 0.867, P = 0.002). Finally, in women during the HOA diet, several markers of of rats consumed 2.5g/kg glucose within 5 minutes, with minor variation in oxidant and infl ammatory stress were decreased (pJNK level in muscle and consumption time between them. Therefore, this technique can be used to serum concentration of interleukin-6 and ferritin). These results provide an reliably administer sweet solutions over a given period of time. In order to as- unprecedented metabolic snapshot of the Mediterranean paradox and boost sess the post-prandial effects of using this technique, we compared glucose, evidence that replacing dietary PA with OA favors cardiometabolic health in insulin and GLP-1 responses evoked by voluntary feeding (VF) to oral gavage a sexually dimorphic manner, but inherent characteristics such as physical (OG) in 12 male Wistar rats (n=6 per group). Each group was administered

fi tness modify the response in women. 2.5g/kg glucose via VF or OG. Animals were killed 30 minutes after adminis- Obesity Supported by: NIH (R01 DK073284, R01 DK082803, RR00109) tration of glucose and blood was collected for analysis of glucose, insulin and glucagon like peptide-1 (GLP-1). Blood glucose (VF: 7.79 ± 0.41 vs OG: 8.25 ± PUBLISHED ONLY 2795-PO 0.51 mmol/L), plasma insulin (VF: 2.60 ± 0.53 vs OG: 3.60 ± 0.39 ng/mL) and Integrated Physiology/ plasma GLP-1 (VF: 29.9 ± 6.2 vs OG: 30.8 ± 4.4 fmol/ml) were similar using both techniques. Therefore, both techniques delivered sugars into the gas- WITHDRAWN trointestinal system equally and induced similar post-prandial responses 30 minutes after delivery. We conclude that voluntary administration of sugar solutions is an effective technique for post-prandial physiological investiga- tions in rats. This technique may retain normal cephalic responses and avoid stress induced by excessive handling. Future work will explore the utility of this technique in glucose tolerance tests and determine effects of this tech- nique on stress and cephalic responses as compared to oral gavage. Supported by: Wellcome Trust, MRC, BBSRC, NIHR

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A695 INTEGRATED PHYSIOLOGY—MACRONUTRIENTCATEGORY METABOLISM AND FOOD INTAKE

2799-PO 2801-PO Toll-Like Receptor 2 Action in the Hypothalamic Regulation of Sick- ness Behavior BYUNG JU LEE, SUNG HO JIN, JAE GEUN KIM, H. MOO KWON, Ulsan, Republic WITHDRAWN of Korea, New Haven, CT Sickness responses such as anorexia and changes in energy metabolism are closely associated with infl ammatory diseases. A number of cytokines and pathogens induce sickness responses through activating infl ammation in the brain. Toll-like receptors (TLRs) are family members of pattern-recog- nition receptors (PRRs) recognizing conserved structural motifs on a wide array of pathogens as well as some endogenous molecules. TLR2, as a PRR, plays a pivotal role in mediating the immune responses in peripheral organs. However, its brain function is largely unknown. This study aims to identify whether acute infl ammation induces sickness responses via activating TLR2 signaling. Intracerebroventricular injection of Pam3CSK4 (Pam3), a synthetic TLR2 ligand, resulted in decreases in food intake, body weight and locomo- tor activity as well as increase in body temperature in the animals. More- over, Pam3 evoked signifi cant increases in the hypothalamic expression of _-melanocyte stimulating hormone, pro-infl ammatory cytokines and prosta- glandin-synthesizing enzymes. However, Pam3-induced sickness behaviors and change of the cytokine expression were less severe in mice lacking with TLR2 and MyD88 expression. A cyclooxygenase inhibitor, indomethacin, as well as a melanocortin receptor (MC3/4R) antagonist, SHU9119, attenuated the effect of Pam3 on anorexia, weight loss, hyperthermia and hypoactiv- ity. These results suggest that TLR2 is a critical mediator of behavioral re- sponses to sickness by regulating infl ammatory system and melanocortin pathway in the brain.

2802-PO Amino Acids and their Metabolites Activate AMP-Activated Protein Kinase Signaling in Hepatocytes YUSUKE ADACHI, IKUE OBARA, YASUSHI NOGUCHI, MARY-ELIZABETH PATTI, Boston, MA, Kawasaki-shi, Japan Recent studies have suggested that plasma levels of amino acids (AA) 2800-PO are associated with risk of T2D, and that dietary amino acids can modulate Altered Levels of Components of the AMPK/mTOR Sensor System whole-body metabolism. AMP-activated protein kinase (AMPK), a key regu- in the Gestational Undernutrition—Low Birth Weight (UN-LBW) lator of energy metabolism in response to various nutritional signals, can Mouse Model of Adiposity exert benefi cial effects on cellular and systemic metabolism. The branched- ARISTIDES LYTRAS, ELVIRA ISGANAITIS, MICHAEL CHEN, WEN KONG, APAR- chain AA leucine can inhibit AMPK via activation of Tor signaling. In this NA SHARMA, YUSUKE ADACHI, HUIJUAN MA, VICENCIA SALES, JOSHUA study, we examined whether additional AA can also modulate AMPK activ- SCHROEDER, ALISON BURKART, MARY-ELIZABETH PATTI, Boston, MA, Athens, ity in mammalian cells and tissues, using high-throughput in-cell Western Greece assays in cultured cells. Increased phosphorylation of AMPK was observed Experimental gestational undernutrition (UN) results in low birth weight in hepatoma H4IIE cells incubated with L-alanine (Ala)(1-10 mM, 1.5-3.7 (LBW) offspring mice, which subsequently develop increased adiposity and fold increases), L-cysteine (Cys) and L-methionine (Met). Ala rapidly acti- progressive glucose intolerance. To test the hypothesis that altered energy vated AMPK over a dosage range of 0.25-10 mM. Moreover, Ala increased and nutrient sensing may contribute to these phenotypes, we assessed the phosphorylation of AMPK by 1.5 fold (p<0.05) in liver in C57BL/6J mice 30 protein expression and/or phosphorylation status of AMPK/mTOR pathway minutes after acute gavage (1.5 g/kg BW). By contrast, Cys and Met (0.5-10 components, including protein phosphatase 2A (PP2A), in liver and adipose mM) had a more gradual onset of AMPK activation (30 minutes), suggesting tissue from10 week old male control (C) and UN-LBW mice. PP2A exerts key that metabolism might be required for activation. Therefore, we measured inhibitory effects on mTOR activity counteracted by amino acids. Mice were activation of AMPK in H4IIE cells treated with Cys and Met metabolites. fasted overnight and administered leucine (CLEU; n=7, UNLEU; n=7) or vehicle Among them, L-homocysteine and L-cysteine sulfi nic acid (CSA) increased (CV; n=9, UNV; n=7), by oral gavage, 1h before sacrifi ce and tissue collection. phosphorylation of AMPK in H4IIE cells. CSA was particularly potent in its Liver PP2A-A (scaffolding subunit) was 25.1% lower (p<0.022) in UN-LBW ability to activate AMPK by 2.5 fold (8-1000 μM, p=0.00001) at doses >5 (n=14) compared to C (n=16) mice, while LEU did not signifi cantly alter PP2A- μM. These data suggest that products of Cys/Met metabolism, including A levels. UN-LBW mice showed a trend for higher phosphorylated mTOR CSA, may mediate AMPK activation. Understanding the cellular mechanisms (p-mTOR; 13.4%, p<0.09) and p-Akt (48.7% p<0.10). Higher UNLEU/UNV ratios, mediating such effects of AA on cellular metabolism and AMPK activity will compared to CLEU/CV, were observed for both p-mTOR (1.17 vs 1.06) and p-S6 be important to elucidate links with regulation of systemic metabolism and (1.36 vs 1.17) values, raising the possibility that UN-LBW mice may be more diabetes risk. responsive to LEU, albeit these differences did not reach signifi cance. No differences in PP2A-A, p-mTOR, p-Akt, or p-S6 were observed in 10 week 2803-PO

Obesity epididymal adipose tissue. However, microarray analysis of adipose tissue The Proportion of Fat in the Diet has a Role in Amygdala Insulin Sen- from 3 week old UN-LBW mice showed trends for reductions in PP2A-A (Pp- sitivity p2r1a; 23.2%, p<0.02, q<0.26 for 60% of the probe sets, while 40% showed STEPHANE BOGHOSSIAN, MIEJUNG PARK-YORK, DAVID A. YORK, Logan, UT PUBLISHED ONLY Integrated Physiology/ no change, Ppp2r1b; 25.5%, p< 0.04, q<0.35), as well as, changes in PP2A-B The anorectic response of insulin administration into the central nucleus (regulatory subunit which confers substrate specifi city) and PP2A-C (cata- of the amygdala (CeA) is lost within 3 days of feeding a high fat diet contain- lytic subunit) isoform mRNAs. We hypothesize that altered PP2A-A, -B and ing 45% kcal fat (fat-45). This study investigates the effects of the propor- -C subunit levels may have differential impact on the tissue-specifi c activity tion of dietary fat on the development of the CeA insulin insensitivity, mainly of the AMPK/mTOR sensor system and hence on energy fl ux and fuel parti- evidenced by the feeding response to CeA insulin administration and on by tioning in UN-LBW vs C mice during postnatal life. the activation of insulin signaling pathways in the CeA area. First, in rats Supported by: European Commission FP7-PEOPLE-2009 MC-IOF-253131 (A.L.) implanted with cannulas aimed at the CeA, the feeding response to CeA insulin (8mU) was tested after 10 days of Low Fat Diet (LFD) feeding. Then the rats were assigned to 4 experimental group; LFD (10% kcal fat), fat-25 (25% kcal fat), fat-45 (45% kcal fat) and fat-60 (60% kcal fat) and tested again for their feeding response to CeA insulin after 24, 72h and 8 days. As

For author disclosure information, see page 797. ADA-Funded Research

A696 INTEGRATED PHYSIOLOGY—MACRONUTRIENTCATEGORY METABOLISM AND FOOD INTAKE expected, there was no response to CeA insulin in the fat-45 and -60 groups 2806-PO after 72h. However, food intake was non signifi cantly decreased after 72h Conditional Knock-in of Sirt1 in AgRP or Pomc Neurons Lead to Lean in the fat-25 group, suggesting a threshold were individual differences could Phenotype Due to Decreased Food Intake and Increased Energy Ex- have a role on CeA response to insulin. Second, in a different experiment, penditure rats implanted with CeA cannulas were fed the same diets and grouped as TSUTOMU SASAKI, MAYUMI SHIMPUKU, OSAMU KIKUCHI, KOSUKE AMANO, described earlier. After 72h, insulin was injected and the rats were sacrifi ced TOMOYA KITAMZUMI, HIROMI YOKOTA-HASHIMOTO, MASAKI KOBAYASHI, TA- 10 minutes after. CeA area were quickly processed for western blot analyses DAHIRO KITAMURA, Maebashi, Gunma, Japan of insulin signaling pathways activation (phosphorylation of AKTser473 and Sirt1 is a longevity gene that is responsible for improved health and life IRS1ser307). Ten minutes after the administration of insulin onto the CeA, span mediated by caloric restriction, and encodes NAD+-dependent deacety- we observed an increase in phosphorylation levels of both AKTser473 and lase. In order to address the contribution of hypothalamic Sirt1 on regulat- IRS1ser307 in the LFD group. The activation of insulin signaling pathways ing long-term energy balance, we developed conditional Sirt1 knock-in mice disappeared in the fat-25, -45, -60 groups. Taken together, our data suggest using Rosa26 promoter and crossed to Pomc-Cre and Agrp-Cre mice. Male that even a small increase in dietary fat is suffi cient to inhibit insulin signal- Pomc-Cre; Rosa26Sirt1-WT conditional knock-in mice (Pomc-Sirt1 KI mice) ing in the CeA and confi rm that the development of CeA insulin resistance is are signifi cantly leaner than wild-type (WT) littermates with less adiposity dependent on the quantity of fat in the diet. Further investigation is needed on normal chow feeding. Food intake was not altered in Pomc-Sirt1-KI mice, to expend these fi ndings. but the mice tend to have higher energy expenditure without any change Supported by: Utah Science Technology and Research Initiative (USTAR) in locomotor activity, suggestive of increased basal metabolic rate. Pomc- Sirt1-KI mice had signifi cantly higher plasma T4 level and higher expression 2804-PO of mitochondrial genes in brown adipose tissue, consistent with higher basal metabolic rate. Pomc-Sirt1 KI mice also have enhanced fasting-induced li- polysis, indicative of higher sympathetic tone. On the other hand, male Agrp- Cre; Rosa26Sirt1-WT conditional knock-in mice (Agrp-Sirt1 KI mice) are also WITHDRAWN lean on normal chow feeding, but present with signifi cantly decreased food intake and trend toward increased energy expenditure without changes in locomotor activity. Conversely, female Agrp-Cre; Rosa26Sirt1-H355Y con- ditional knock-in mice, which expresses enzyme-dead Sirt1-H355Y mutant, tend to gain more weight with signifi cantly increased food intake. Energy expenditure was not different from wild-type mice in Agrp-Sirt1-H355Y knock-in female mice. Therefore, Sirt1 in both Pomc neurons and Agrp neu- rons contribute to the regulation of energy balance. Sirt1 in Pomc neurons have stronger infl uence on energy expenditure, whereas the effect of Sirt1 in Agrp neurons is stronger on food intake.

2807-PO Ghrelin Action on NPY Neurons: Direct versus Indirect? HIROSHI HASHIGUCHI, ZHENYU SHENG, VANESSA H. ROUTH, JOSEPH BRYAN, Seattle, WA, Newark, NJ How ghrelin activates NPY neurons is unclear. Data on isolated neurons show direct action, while CNQX inhibits ghrelin (GHR) induced action po- tentials (APs) in NPY neurons in brain slices implying a presynaptic effect. Ca2+-imaging shows direct GHR action on 100% of isolated ‘green’ NPY-GFP neurons. Evaluation by whole cell current clamp recording in slices shows ~50% of NPY-GFP neurons are glucose-inhibited (GI); GHR depolarized the majority of NPY-GFP neurons increasing their AP frequency. Multiple chan- nels may be involved; GHR both decreased and increased input resistance consistent with channel opening and closure, respectively. Tetrodotoxin (TTX) blocked GHR effects on membrane potential and increased IR con- sistent with presynaptic action in at least some cases. In isolated neurons inhibition of AMPK blocked, while AICAR mimicked GHR action. Inhibiting CAMKK2, a Ca2+-dependent AMPK activator, with STO-609 blocked GHR action. Activation of adenylyl cyclase elevated [Ca2+]c like GHR, an effect 2805-PO partially blocked by PKA inhibition. Cl--free media elevated [Ca2+]c, while GABA inhibited GHR action consistent with membrane hyperpolarization. Ca2+ channel blocker studies implicated R-type, but not L-, N- or T-type channels in GHR action. CNQX had no effect on direct GHR actions on iso- WITHDRAWN lated neurons, consistent with no synaptic contacts. TTX also had no effect, but lowering [Na+]o did block GHR action implicating non-selective cation channels. KATP channel modulators gave paradoxical results: diazoxide, an agonist, had no effect on GHR action, but did elevate [Ca2+]c without GHR, while glibenclamide, an antagonist, reduced [Ca2+]c in the presence of GHR.

The pharmacologic results are consistent with the stress of isolation up-reg- Obesity ulating SUR1-regulated NCCa-ATP channels, non-selective cation channels activated by Ca2+ when [ATP]c is low, known to be induced under ischemic PUBLISHED ONLY

conditions. The data suggest caution when interpreting results from isolated Integrated Physiology/ hypothalamic neurons. Supported by: NIH DK081538 (V.H.R.), DOD W81XWH-11-1-0305 (J.B.)

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A697 INTEGRATED CATEGORYPHYSIOLOGY—MUSCLE

2808-PO 2810-PO Antioxidant Activity of Kawaratake (Trametes Versicolor) against Dietary Pattern and Microelement in China Diabetic Patients the Oxidized Low-Density Lipoprotein HONG WU, DAJIN ZOU, Shanghai, China MASUMI KAMIYAMA, TAKAO ANDO, MINORU TANAKA, MASAHIKO FUJII, Different studies in adults have shown that dietary pattern and microele- TAMAMI IWAMOTO, TOSHIMI KIDO, KAZUO KONDO, Tokyo, Japan, Saitama, ment play a role in diabetic pathogenesy. It was to determine the associa- Japan tion between dietary pattern plus microelement and gender, BMI, Diabetic Trametes versicolor is known as Kawaratake and its immune-enhancing type, diabetic treatment protocols, in Diabetic patients in China.. Over 1500 activities have been extensively studied in Japan. In 1977, an extract of diabetic patients (668 males) from 30 hospitals in China, were examined cultured mycelia of Kawaratake, protein-bound polysaccharide preparation and questionaired between April and September 2010. Dietary pattern (Krestin) was approved by the Japanese Ministry of Health and Welfare to was collected by questionnaire survey. Vitamins and Microelements were use for cancer treatment as the fi rst polysaccharide antitumor drug from determined including calcium, iron, zinc, selenium, cuprum, manganece, mushroom. In addition to various mechanisms of action, the possibility of an- magnesium,and natrium. Body max index (BMI), Glycosylated hemoglobin tioxidant activity of Kawaratake has been shown. However, detailed mecha- A1C (HbA1C) , Fasting blood glucose (FBG) and postprandial glucose (PBG) nism and useful components remain to be clarifi ed. Fruit body of Kawaratake were also determined. Over 303 (20.27%) diabetic patients were obese (OB), was extracted with 95% ethanol. Also, we prepared molecular weight (MW) and 613 (41.0%) overweight (OW) using centers for disease control (CDC) fractions of Krestin (MW 3,000-20,000, MW 20,000-100,000, and MW norms. Dietary pattern of OB/ OW patients was signifi cantly different from grater than 100,000). We examined the concentration of total polyphenols normal weight (NW) patients (p < 0.05). Vitamin A, Vitamin B2, and Vitamin and amino acid of Krestin itself. Then, we investigated DPPH (1,1-diphenyl- C levels in OB /OW patients were lower than NW patients, respectively. (p 2-picrylhydrazyl) radical scavenging activity and antioxidant activity using = 0.0077, p=0.0071, p=0.0148). Dietary pattern was also different between oxidized low-density lipoprotein (LDL). We detected high concentration of type 1 diabetes and type 2 diabetes patients (p < 0.05). Vitamin A, Vitamin total polyphenols and some amino acids that have antioxidant properties B2, and Vitamin C level was higher in type 1 diabetes (p = 0.0104, p=0.0043, (14% glutamic acid, 9% alanine, 2% methionine, 7% phenylalanine, and 2% p=0.05), while Vitamin E and Vitamin B1 showed no difference between histidine). DPPH radical scavenging activity of Kawaratake solution, and these two groups. Our results suggest that the prevalence of OW/OB is MW fractions and unfractionated solution of Krestin was not detected. higher in diabetic patients in China. Dietary pattern and Vitamins were as- However, Kawaratake solution, and MW fractions and unfractionated solu- sociated with diabetic type and weight difference. Microelements including tion of Krestin showed the signifi cant inhibition of LDL oxidation (ratio of calcium, iron, zinc, selenium, cuprum, manganece, magnesium, and natrium antioxidant activity against control as 1; Kawaratake solution: 1.44, MW have no signifi cant effects on diabetic patients. 3,000-20,000: 1.73, MW 20,000-100,000: 1.36, MW grater than 100,000: 1.53, and unfractionated Krestin solution: 2.06). Our results suggest that the components of Kawaratake maintain LDL resistance to oxidation in spite of INTEGRATED PHYSIOLOGY—MUSCLE no direct activity of radical scavenging. In conclusion, intake of Kawaratake solution is useful for the reduction of oxidized low-density lipoprotein. 2811-PO Loss of Akt Signaling in Skeletal Muscle Causes Age-Dependent Insulin Resistance With Progressive Sarcopenia 2809-PO TOSHIHIRO UMEHARA, KOHJIRO UEKI, KAZUMA KANEKO, TAKAYOSHI SASAKO, MOTOHARU AWAZAWA, YUKIKO OKAZAKI, C.R. KAHN, MORRIS J. BIRNBAUM, TAKASHI KADOWAKI, Tokyo, Japan, Boston, MA, Philadelphia, PA Aging is one the major risk factors of type 2 diabetes associated with age-dependent decreased insulin sensitivity in skeletal muscle, at least in WITHDRAWN part. Indeed, aged people often show decreased skeletal muscle volume, the major site of glucose disposal, and increased adipose tissue, namely sarcopenia. Although age-dependent changes in insulin/IGF-1 signaling, mitochondrial functions and autophagy are thought to be involved in this process, the precise mechanism still remains unclear. Since Akt is the key signaling mediator of insulin /IGF-1 signaling and appears to regulate mi- tochondria and autophagy, in the current study, to assess the Akt role in the context of age-dependent insulin resistance in skeletal muscle, we gen- erated skeletal muscle specifi c Akt1 and Akt2 double knockout mice using MLC1f-Cre mice (MLC1f-DKO mice). Different from the Akt1 and Akt2 double knockout mice using MCK-Cre mice which die within 5 weeks after birth due to the heart failure, MLC1f-DKO mice grow normally and show normal glu- cose tolerance until 8 weeks of age despite the complete shutdown of sig- naling downstream of Akt. Surprisingly, insulin-stimulated glucose uptake is preserved even in the isolated skeletal muscle of these mice. However, MLC1f-DKO mice develop systemic insulin resistance in an age-dependent fashion with decreased skeletal muscle vaolume. In muscle of MLC1f-DKO mice, genes regulating fatty acid oxidation and oxidative phosphorylation are signifi cantly decreased and mitochondria DNA copy number is also de- creased. Moreover, insulin-stimulated glucose uptake is impaired in isolated skeletal muscle of the aged MLC1f-DKO mice. These data suggest that Akt

Obesity in skeletal muscle plays a pivotal role in maintaining skeletal muscle vol- ume and functions associated with insulin sensitivity against aging, and that decreased insulin action in skeletal muscle may exacerbate age-dependent PUBLISHED ONLY

Integrated Physiology/ decrease in insulin sensitivity by accelerated sarcopenia due to decreased Akt signaling.

2812-PO BRSK1 Promotes Glucose Uptake in L6 Myotubes and Mouse Skel- etal Muscle KAZUHIRO NAKANO, XU YAN, AN DING, MICHAEL HIRSHMAN, LAURIE GOOD- YEAR, Boston, MA AMPK and its upstream kinase LKB1 regulate glucose uptake in skeletal muscle. Brain specifi c kinase 1(Brsk1) is an AMPK family member activated by LKB1 via Thr189 phosphorylation and is required for neuronal polariza-

For author disclosure information, see page 797. ADA-Funded Research

A698 INTEGRATED CATEGORYPHYSIOLOGY—MUSCLE tion in mammals. Three Brsk1 isoforms have been reported in brain (long cular blood fl ow (MBF) were determined at baseline, 240, and 360 min us- form, short form, S1 form), whereas there have been no studies of Brsk1 ing contrast-enhanced ultrasound. Candesartan treatment acutely recruited expression and function in skeletal muscle. We used L6 muscle cells and microvasculature in both skeletal and cardiac muscle by increasing MBV adult mouse skeletal muscle to study the expression and function of Brsk1 (~70%, p<0.03 and ~40% p<0.02 respectively) and MBF (~60% and ~40%, in vitro and in vivo. Of the three isoforms of Brsk1, mRNA and protein analy- p<0.03 for both) without altering MFV. Insulin increased cardiac MBV (~50%, ses showed that L6 cells and skeletal muscles abundantly express the short p<0.03) and MBF (p<0.02) but the increase in skeletal muscle was not sta- form. Short form Brsk1 expression was increased during differentiation of L6 tistically signifi cant (~60%, p=0.14). Superimposing insulin infusion 240 min cells from myoblasts to myotubes. To study the function of short form Brsk1 after candesartan administration for two hours did not further recruit micro- in muscle, wild type and mutant (Thr189Ala) Brsk1 were overexpressed in L6 vasculature in skeletal muscle. Candesartan pre-treatment did not change myotubes (4-fold) and shRNA was used to knockdown Brsk1 (34% decrease). insulin-mediated whole body glucose disposal. Cardiac MBV changes cor- Wild type Bsk1 overexpression had no effect on basal glucose uptake, but related positively with plasma renin activity at 240 min (a = 0.728, p=0.007). increased insulin-stimulated (30nM, 30min) glucose uptake by 51% vs. emp- In conclusion, acute AT1R blockade with candesartan recruits skeletal as ty vector control cells (p<0.05). Although mutant Brsk1 expression had no well as cardiac muscle microvasculature in healthy humans without altering effect on glucose uptake, Brsk1 knockdown decreased basal glucose uptake insulin-mediated whole body glucose disposal. This may contribute to the by 21% (p<0.05), and insulin-stimulated glucose uptake tended to decrease observed improvement in cardiovascular outcomes and insulin sensitivity compared to scrambled shRNA (23%, p=0.09). To study the role of Brsk1 in seen in subjects who receive prolonged AT1R blockers. vivo, short form Brsk1 was expressed in tibialis anterior (TA) muscles by in Supported by: NIH vivo gene injection and electroporation. Similar to L6 cells, wild type Brsk1 overexpression tended to increase insulin-stimulated glucose uptake in TA 2815-PO (77%, p=0.09), with no effect of mutant Brsk1. In summary, overexpression of the short form of Brsk1 affects insulin-stimulated glucose uptake in L6 myotubes and skeletal muscle, and knock down of Brsk1 in L6 myotubes WITHDRAWN decreases basal and insulin-stimulated glucose uptake. The short form of Brsk1, an AMPK related kinase, is highly expressed in skeletal muscle and may function in the regulation of insulin action.

2813-PO 2816-PO MicroRNA-494 Regulates Mitochondrial Biogenesis in Skeletal Muscle through Mitochondrial Transcriptional Factor A (mtTFA) and Forkhead Box j3 (Foxj3) KATSUTARO MORINO, HIROTAKA YAMAMOTO, YOSHIHIKO NISHIO, SATOSHI WITHDRAWN UGI, TAKESHI YOSHIZAKI, OSAMU SEKINE, ATSUNORI KASHIWAGI, HIROSHI MAEGAWA, Otsu, Japan, Kagoshima, Japan Skeletal muscles require a large amount of energy, and the number and function of mitochondria within skeletal muscle cells are related to their individual metabolizing capacity; thus, any dysfunction may result in meta- bolic disease. Recent reports suggested a relationship between mitochon- drial dysfunction and diabetes. MicoRNAs (miRNAs) are important post- transcriptional regulators of various biological pathways. In this study, we focused on the role of miRNAs during mitochondrial biogenesis in skeletal muscle. First, Micro-array based screening was performed to fi nd novel miR- NAs related with skeletal muscle differentiation. We have found that the ex- pression of miR-494 markedly decreased in murine myoblast C2C12 cells dur- ing myogenic differentiation, accompanied by an increase in mitochondrial DNA. Also, the expression of predicted target genes for miR-494, including mitochondria transcriptional factor A (mtTFA) and Forkhead box j3 (Foxj3), was post-transcriptionally increased during myogenic differentiation. Inhibi- tion of miR-494 by a specifi c antisense resulted in increase in mitochondrial content and upregulated levels of mtTFA and Foxj3 at the protein level. In addition, overexpression of miR-494 decreased protein expression of mt- TFA and Foxj3 in C2C12 cells. A 3’UTR reporter assay revealed that miR-494 inhibition directly upregulated the luciferase activity of mtTFA and Foxj3. Furthermore, the miR-494 content signifi cantly decreased after endurance exercise in C57BL/6J mice, accompanied by an increase in expression of the mtTFA and Foxj3 proteins. These results suggest that miR-494 regulates mi- tochondrial biogenesis by downregulating mtTFA and Foxj3 during myocyte differentiation and skeletal muscle adaptation to physical stimulation.

2814-PO Candesartan Acutely Recruits Skeletal and Cardiac Muscle Micro- vasculature in Healthy Humans 2817-PO Obesity MATTHEW A. SAUDER, JIA LIU, LINDA JAHN, DALE E. FOWLER, WEIDONG CHAI, Peroxisomal Catalase Restricts Accumulation of Hydrogen Perox- ZHENQI LIU, Charlottesville, VA ide Emitted by Mitochondria in Human Skeletal Muscle PUBLISHED ONLY Angiotensin II type 1 receptor (AT1R) blockade reduces cardiovascular VLADIMIR B. RITOV, ELIZAVETA V. MENCHIKOVA, PAUL M. COEN, GIOVANNA Integrated Physiology/ incidents and new onset diabetes. We recently reported that in laboratory DISTEFANO, NICOLE HELBLING, JASON NG, MAJA STEFANOVIC-RACIC, FRED- animals AT1R tone restricts muscle microvascular blood volume (MBV) and ERICO TOLEDO, BRET H. GOODPASTER, Pittsburgh, PA decreases muscle insulin delivery and glucose use. To examine whether Recent observations indicate that mitochondrial derived oxidative acute AT1R blockade alters skeletal and cardiac microvascular perfusion in stress contributes to skeletal muscle insulin resistance in obesity or type 2 humans, we studied 8 healthy, overnight-fasted young adults thrice in ran- diabetes. We tested hypothesis that accumulation of mitochondrial derived dom order. In study 1, subjects received candesartan (32 mg) orally at time H2O2 from permeabilized muscle fi ber bundles is dependent on peroxisomal 0. In study 2, subjects received placebo at time 0 and a 1 mU/min/kg eugly- catalase activity. Permeabilized fi ber bundles were prepared from vastus cemic insulin clamp from time 240 to 360 min. In study 3, subjects received lateralis biopsies obtained from morbidly obese subjects at 1-3 months post- candesartan at time 0 and insulin infusion from 240 to 360 min. Skeletal bariatric surgery. A “non-invasive” Amplex red HPLC based techniques for and cardiac muscle MBV, microvascular fl ow velocity (MFV) and microvas- analysis of accumulation of H2O2 in small volume (~100 μl) was developed.

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A699 INTEGRATED CATEGORYPHYSIOLOGY—MUSCLE

o Accumulation of H2O2 was determined at 37 C in the presence of succinate whether this gene transfer technology can be applied to study physiological plus antimycin A and in the absence of Amplex red and peroxidase to spare consequences, we selected GLUT4 as a target for shRNA mediated knock- the generated H2O2 before it can reach peroxisomal catalase. The fi ber bun- down due to its well characterized role in facilitating the rate limiting step of dles demonstrate succinate dehydrogenase (SDH) activity (1.03±0.16 U/g insulin induced glucose uptake into skeletal muscle. We are able to demon- wet weight at 30oC; n=15) that linearly depends on the fi ber bundles wet strate that 4 weeks after administration of AAV8 carrying GLUT4 shRNA into weight (R2=0.87; P<0.05; n=15), indicating good accessibility of fi ber interior the tibialis muscle of mice GLUT4 mRNA and protein expression can be re- for succinate. The fi ber bundles demonstrate tightly bound catalase activity pressed by 40-60%. Functionally, this reduction in GLUT4 protein expression that is active at low concentration of H2O2 (~1 μM) and can be blocked com- was associated with a signifi cant 30-40% reduction in insulin-stimulated pletely by low concentrations of sodium azide. These indicate the presence 2-deoxyglucose tracer uptake ex vivo. In conclusion, AAV8 mediated gene of peroxisomes in the bundles. At normal oxygen tension the succinate sup- transfer provides a suitable tool to study the functional role of proteins upon ported generation of H2O2 by the fi ber bundles is non-linear in time, reaching long-term manipulation of their expression in skeletal muscle. a peak concentration 2.18±0.70 μM (n=21) for 30 min of incubation. In the presence of sodium azide (50 μM) the time dependence of succinate sup- ported accumulation of H2O2 in the fi ber bundles incubation medium became 2820-PO absolutely linear at least for 45 min of incubation. These data indicate that Enhanced NAD Synthesis in Skeletal Muscle is Insuffi cient to Pro- peroxisomal catalase restricts accumulation of mitochondria derived H2O2 mote Mitochondrial Biogenesis or Protect Against the Metabolic in human skeletal muscle. This novel data provide valuable insight into the Consequences of High Fat Diet kinetics of H2O2 accumulation in human muscle and suggest an important DAVID W. FREDERICK, JAMES G. DAVIS, BEAMON AGARWAL, EIKO NAKAMA- role for peroxisomal catalase activity. RU-OGISO, JOSEPH A. BAUR, Philadelphia, PA Recent work suggests that skeletal muscle plays a critical role in the regulation of whole body energy balance during calorie restriction (CR) by 2818-PO increasing fatty acid oxidation and insulin sensitivity in a sirtuin 1 (SirT1)-de- pendent manner. NAD-dependent deacetylation of peroxisome proliferator- activated receptor gamma coactivator 1-alpha (PGC-1a) by SirT1 has been proposed to underlie numerous metabolic responses to CR, some of which WITHDRAWN might be reproduced by strategies that increase the availability of NAD. Interestingly, the expression of nicotinamide phosphoribosyltransferase (Nampt), the rate-limiting enzyme in the mammalian NAD salvage pathway, increases in response to both nutrient deprivation and exercise. We tested whether expression of Nampt under control of a muscle creatine kinase pro- moter (MCK-Nampt) would partially mimic the CR state and protect against the metabolic impairments associated with a sedentary lifestyle or ad libi- tum feeding. Preliminary characterization indicates that skeletal muscle in MCK-Nampt mice does not phenocopy prominent transcriptional features of muscle from mice under chronic CR. Despite a 35% increase in intramuscular NAD content, we observed no shifts in respiratory exchange ratio, markers of PGC-1a activity, or mitochondrial content. Additionally, when challenged with a high fat diet (60% calories from fat), MCK-Nampt mice demonstrate a complete rescue of the diet-induced decline in intramuscular NAD, but exhib- it no protection against the development of obesity or glucose intolerance. Ongoing experiments aim to measure the intramuscular abundance of NADH and nicotinamide, as well as acetylation states of SirT1 substrates. Our re- sults suggest that limited nutrient availability invokes additional signals that must coincide with elevated NAD to elicit the metabolic reprogramming of skeletal muscle during CR.

2821-PO AKT Interacts With Smad3 Determining Myogenesis vs. Fibrosis in Injured Muscles YANJUN DONG, RONALK LAKHIA, YANLAN DONG, WILLIAM E. MITCH, LIPING ZHANG, Houston, TX Impaired IGF1 signaling causes weakness, muscle atrophy and reduced muscle regeneration. To explore how decreased IGF1 signaling impairs re- generation, we created satellite cell-specifi c IGF1R KO mice (crossing fl oxed- IGF1R x MyoD-cre mice). Responses to cardiotoxin-induced muscle injury were compared in IGF1R KO and Floxed-IGF1R mice. After 5, 7 or 14 days of 2819-PO muscle injury in IGF1R KO mice, we found fewer and smaller new myofi bers Long-Term GLUT4 Knockdown by AAV8 Mediated Gene Transfer in (i.e., cells with central nuclei; p<0.05) with decreased mRNA and protein of Skeletal Muscle of Mice—A New Gene Transfer Technology Ap- MyoD and myogenin (satellite cell proliferation and differentiation markers plied to Study Insulin Sensitivity respectively); fewer Brdu incorporation in satellite cells; increased mRNA of Obesity SEBASTIAN KREUZ, STEPHAN GLUND, GERD LUIPPOLD, CORINNA SCHOELCH, TGF-`, _-smooth muscle actin, collagen I (p<0.05); increased p-Smad3 and ROBERT AUGUSTIN, Biberach a.d. Riss, Germany trichrome staining in injured muscles. Thus, impaired IGF1 signaling in satel- Studying the functional role for particular genes of interest and their im- lite cells suppresses muscle regeneration but increases fi brosis. To identify PUBLISHED ONLY Integrated Physiology/ pact on skeletal muscle physiology in rodents is currently limited to conven- how impaired IGF1 infl uences TGF-` signaling, we infected C2C12 myoblasts tional or tissue-specifi c gene manipulation, a laborious endeavour, which can with adenoviruses expressing either constitutively active Akt (Myr-Akt) or be e.g. limited by developmental adaptation of animal physiology. Alterna- dominate-negative Akt (Akt-AAA). In response to Myr-Akt, p-Smad3 fell tive approaches applying in vivo electrotransfer or adenoviral transduction sharply. But, in response to Akt-AAA p-Smad3 rose. An interaction between are hampered by short-term effects on gene expression limiting their use Akt and Smad3 was confi rmed in C2C12 cells transfected with Flag-Smad3: to investigate long-term physiological consequences. Here we show that Akt was present in immune complexes with anti-Flag. IGF1 treatment in- in skeletal muscle of mice long-term alterations of local gene expression creased this association while TGF-`1 treatment blocked it. Thus, the as- by RNAi can be achieved by Adeno-associated virus (AAV) 8 mediated gene sociation of Akt with Smad3 impairs TGF-` signaling, promoting satellite cell transfer. Initial validation of the technology was provided using AAV8 vec- proliferation and differentiation. tors expressing GFP or Luciferase and subsequent monitoring of transgene expression in injected muscles by in vivo imaging..In order to investigate

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A700 INTEGRATED CATEGORYPHYSIOLOGY—MUSCLE

Impaired IGF1 signaling in satellite cells promotes TGF-` signaling and fi - brosis (Figure)

2824-PO Imbalance of Mitochondria Dynamics Contributes to Mitochondrial Dysfunction and Metabolic Deterioration in Skeletal Muscle HUEI-FEN JHENG, PEI-JANE TSAI, SYUE-MAIO KUO, LI-HUA KUO, CHERNG-SHY- ANG CHANG, IN-JEN SU, CHUANG-RUNG CHANG, YAU-SHENG TSAI, Tainan, Taiwan, Hsinchu, Taiwan Mitochondrial dysfunction in skeletal muscle has been implicated in the development of insulin resistance and type 2 diabetes. Considering the im- portance of mitochondrial dynamics in mitochondrial and cellular functions, we hypothesized that obesity and excess energy intake shift the balance of mitochondrial dynamics, further contributing to mitochondrial dysfunc- tion and metabolic deterioration in skeletal muscle. First, we revealed that Supported by: Satellite Health, Harold Selzman, and NIHR37 excess palmitate (PA), but not hyperglycemia, hyperinsulinemia, or elevated TNF_, induced mitochondrial fragmentation and increased mitochondria- associated Drp1 and Fis1 in differentiated C2C12 muscle cells. This frag- 2822-PO mentation was associated with increased oxidative stress, mitochondrial ANT1 Carbonylome, The Gate Keeper of Mitochondrial Basal Proton depolarization, loss of ATP production, and reduced insulin-stimulated glu- Leak and its Role in Insulin Resistance Etiology cose uptake. Both genetic and pharmacological inhibition of Drp1 attenu- JEANINE M. CORDOVA, NATALIE LEFORT, SARAH KUZMIAK, WAYNE WILLIS, ated PA-induced mitochondrial fragmentation, mitochondrial depolarization, CARRIE SHARROF, PAUL LANGLAIS, LAWRENCE J. MANDARINO, Scottsdale, AZ, and insulin resistance in C2C12 cells. Furthermore, we found smaller and Tempe, AZ shorter mitochondria and increased mitochondrial fi ssion machinery in the Excessive reactive oxygen species (ROS) and subsequent oxidative stress skeletal muscle of both genetic and diet-induced obese mice. Inhibition of (OS) have been implicated in the etiology of insulin resistance (IR). Because mitochondrial fi ssion improved muscle insulin signaling and systemic insulin protonmotive force and the oxidation-reduction energy down the electron sensitivity of obese mice. Our fi ndings indicated that aberrant mitochondrial transport chain are the energetic precursors to higher 6GATP in IR, mito- fi ssion is causally associated with mitochondrial dysfunction and insulin re- chondria (Mito) from IR muscle maintain higher thermodynamic driving force sistance in skeletal muscle. Thus, disruption of mitochondrial dynamics may and ROS production rates. It has been proposed that under these conditions, underlie the pathogenesis of muscle insulin resistance in obesity and type Mito compensate for increased protonmotive force by raising proton leak, 2 diabetes. primarily by way of oxidative changes in Adenine Nucleotide Translocase Supported by: NSC98-2320-B-006-009-MY3 1 (ANT1), which serves as the ADP/ATP carrier and also a main proton leak contributor. OS leads to post-translational modifi cations (PTM) of proteins and lipids, including protein carbonylation, a spontaneous, mostly irrevers- 2825-PO ible (PTM) that occurs by a free radical reacting with an amino acid side Protein Breakdown Rates in Obese Subjects With Metabolic chain. We previously reported functional lysine acetylation sites in ANT1. Syndrome Here we report the fi rst carbonylome of human skeletal muscle ANT1 and re- LETICIA CASTILLO, SUSAN M. HUTSON, MARIA A. RAMOS-ROMAN, ELIZABETH late these changes to Mito ROS production, respiration and IR. To accomplish J. PARKS, Dallas, TX, Blacksburg, VA this, we used mass spectrometry and normalized spectral abundance factor Obesity is thought to increase whole-body protein breakdown (PB), but (NSAF) quantifi cation with Mito isolated from muscle biopsies of 3 lean, in- the interrelationships between insulin resistance, body composition, plasma sulin sensitive (IS) and 3 obese, IR nondiabetics. Glucose clamps were used amino acid (AA) concentration (conc), and PB have not been explored. The to measure insulin action (Rd) Rd was 4.1±0.4 vs 9.5±0.5 mg/(kg.min) in IR present study measured PB in insulin resistant (IVGTT SI mean ± SD: 2.6 ± -4 -1 vs IS, respectively. We detected 21 unique carbonylated residues on ANT1, 1.5 10 * min per μU/mL), obese Hispanics (Hisp) and African Americans 2 including; 12 hydroxylation, 3 oxidation, 2 oxolactone, 1 carbidomethyl, 1 (AA) (13 females, 9 males, age 47 ± 9y, BMI 34.8 ± 6.5 kg/m ). PB was as- 2 cystic acid and 2 kynurenin. Abundance of ANT1 hydroxylation determined sessed by continuous IV infusion of L-( H3) leucine, the rate of appearance 13 by NASF was inversely related to state 3 respiration utilizing pyruvate and of plasma free fatty acids (RaFFA) by C4-palmitate infusion, followed by malate as fuel (R2=0.68) and directly related to ROS production (R2=0.40). GC/MS. Plasma AA concs were measured by HPLC, energy expenditure (EE) We conclude that site-specifi c oxidation of ANT1 may lower maximum res- by indirect calorimetry, and body composition by DEXA. Protein intake was piration rates and could be a compensatory effort to reduce protonmotive controlled for 1 wk before the study as all food was provided by the clini- force and lower ROS production in insulin resistant muscle. cal research center to maintain constant energy intake. Activity level was Supported by: NIH/NIDDK limited the day before the study. After consuming a 6 PM dinner, subjects Obesity fasted until noon the next day, and then consumed a standardized meal (41g fat, 100g CHO). The ethnic groups did not differ in their conc of any AA, nor

2823-PO PUBLISHED ONLY PB rates - expressed in units of mmol/h (7.1 ± 1.6 vs 6.5 ± 1.6 for Hisp and AA, Integrated Physiology/ respectively, P=0.70) or mmol/kg LBM/h (134 ± 17 vs 123 ± 17, P=0.15). Higher PB was associated positively with elevated lean body mass (LBM, r=0.846), WITHDRAWN higher fasted EE (r=0.905), fed EE (r=0.848) and SI (r=0.601, P<0.004 for all); these characteristics were also associated with a lower conc of a subset of EAA (Tyr, Ile, Val, Lys). PB correlated negatively with trunk fat mass (r=- 0.446, P=0.04), fed-state RaFFA (r=-0.655, P=0.02), and postmeal FFA conc (r=-0.571, P=0.007). Within this group of insulin resistant subjects, those with a high PB had elevated LBM and EE. These characteristics may have positive metabolic implications. Supported by: NIH (RL1 DK 081187)

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A701 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES

2826-PO vs. (17.76 [11.51-24.51] ng/ml, P < 0.001). In addition, N-terminal pro-brain Pre-Training Higher Phosphorylation of Muscle IRS-1 Ser337 and natriuretic peptide, a well-established powerful risk marker in CHF, was in- Ser636 is Associated With Lack of Improvement of Insulin Resis- dependently and positively correlated with A-FABP (Standardized ` = 0.340, tance after Strength Training in the Metabolic Syndrome P < 0.001) after adjusting for confounding factors. Each echocardiographic CHARLES A. STUART, MELANIE P. MCCURRY, MARY E. HOWELL, MARK A. parameter, especially left ventricular ejection fraction, was independently SOUTH, ANDREW S. LAYNE, MICHAEL W. RAMSEY, MICHAEL H. STONE, John- associated with A-FABP (all P < 0.01). Multiple logistic regression analysis son City, TN demonstrated that A-FABP concentration was an independent risk factor Ten non-diabetic volunteers with the metabolic syndrome underwent for CHF (OR = 7.951, 95%CI: 2.324-27.210, P = 0.001). In conclusion, our re- eight weeks of supervised strength training with no weight loss permitted. sults provide the fi rst clinical evidence that A-FABP is closely associated Even though body composition, strength, endurance, blood pressure, and with CHF, and suggest that increased A-FABP may be causally related to the plasma lipids improved, insulin resistance did not change as quantifi ed by pathogenesis of heart dysfunction in humans. euglycemic clamp studies. Signifi cant increases in both insulin receptor and Supported by: The National 973 Program (2011CB504001) GLUT4 expression in skeletal muscle suggested that there was a persistent obstacle in the intracellular pathway mediating insulin-stimulated transloca- 2830-PO tion of GLUT4 in muscle. We have previously shown that persons with the metabolic syndrome have a higher proportion of fast-twitch muscle fi bers and overexpression of activated mTOR. In pre-training samples, we found WITHDRAWN no signifi cant difference between metabolic syndrome subjects and control subjects in the expression of muscle insulin receptor substrate-1 (IRS-1). Immunoblot estimates showed a 18% lower phosphorylation of Tyr896 in metabolic syndrome subjects. IRS-1 serine phosphorylation was compared at four sites. Ser337 phosphorylation was four-fold higher and Ser636 was 2831-PO nearly three-fold higher in the metabolic syndrome, whereas Ser794 and Expression and Distribution of PANcreatic-DERived Factor (PAN- Ser1101 phosphorylation were not different. Ser636 has been shown to be a DER) in Human Gastric and Intestinal Tissues site where phosphorylation occurs in response to mTOR and Ser337 is an ac- SHARI L. MOAK, MARK G. ATHANASON, GRACE C. DOUGAN, MELANIE N. ceptor site for phosphate when GSK-3 is activated. Ser794 phosphorylation KUEHL, WHITNEY A. DANSE, CLAUDIA E. ROBERT-COOPERMAN, BRANT R. is associated with AMPK activation and Ser1101 is phosphorylated by PKC- BURKHARDT, Tampa, FL theta. These data suggest that a major part of the insulin resistance seen in PANcreatic-DERived factor (PANDER, FAM3B) is a 235-amino acid se- the metabolic syndrome is due to the baseline status of serine phosphoryla- creted protein that has been previously characterized to be predominantly tion of IRS-1. It is likely that one or more serine sites with excess phospho- expressed in the endocrine pancreas. Recent animal models have demon- rylation will impair the scaffolding function of this protein and prevent the strated that PANDER regulates glycemic levels via interaction with the liver normal assembly of the components of PI3-kinase, diminishing this critical by inhibition of insulin signaling and subsequently increasing hepatic glu- step in insulin-stimulated glucose transporter translocation in muscle. cose production. Much of the previous work has only focused on PANDER ex- pression in the endocrine pancreas with no evaluation of other tissues that 2827-PO impact glucose homeostasis. To address this, we investigated the expres- sion and distribution of PANDER in both the human and mouse intestine and stomach. Various human tissue lysates were obtained and western analysis WITHDRAWN was performed with an anti-PANDER polyclonal antibody revealing robust PANDER expression in the following human tissues as listed by decreas- ing degree of expression: (1) stomach, (2) pancreas, (3) duodenum, (4) small intestine, and (5) colon. Furthermore, formalin-fi xed, paraffi n-embedded human, rat, and mouse small intestine were evaluated by confocal immu- 2828-PO nofl uorescence microscopy (IFM) and demonstrated distribution of PANDER primarily to the apical striated border membrane with no expression within the basal regions for all 3 species. Further confi rmation of this distribution WITHDRAWN was achieved by similar results obtained by immunohistochemical analysis of PANDER probed formalin-fi xed, paraffi n embedded sectioned murine tis- sues obtained from the duodenum, jejunum, and ileum of C57BL/6 mice. RT-PCR analysis of small intestinal murine tissues revealed the strongest expression of PANDER in the duodenum. PANDER mRNA was not detected in the GLUTAG GLP-1 enteroendocrine cell line as determined by RT-PCR which INTEGRATED PHYSIOLOGY—OTHER HORMONES is consistent with the PANDER non-incretin distribution as identifi ed by IFM. In summary, here we present the fi rst report demonstrating robust PANDER 2829-PO expression and distribution in the small intestine and stomach. Circulating Adipocyte-Fatty Acid Binding Protein Levels are Inde- pendently Associated With Congestive Heart Failure 2832-PO MI ZHOU, MINGYA LIU, YUQIAN BAO, ZHIYONG XU, HUATING LI, HAO ZHANG, WEI ZHU, JIALIANG ZHANG, AIMIN XU, MENG WEI, WEIPING JIA, Shanghai, China, Pokfulam, Hong Kong WITHDRAWN It is well established that obesity is one of the most common risk factors Obesity for cardiovascular diseases, and is directly associated with an increased risk of heart failure. Animal based studies suggested that a predominant protein secreted by adipose tissue, adipocyte fatty acid-binding protein (A-FABP) PUBLISHED ONLY Integrated Physiology/ plays a key role in obesity-related insulin resistance, infl ammation and ath- 2833-PO erosclerosis. We sought to investigate the association of A-FABP with con- gestive heart failure (CHF) in Chinese subjects. Serum A-FABP levels were measured in 252 CHF patients and 261 age, gender, and body mass index WITHDRAWN (BMI) matched non-CHF subjects. Echocardiography was performed on each patient. The severity of CHF was determined by the New York Heart Asso- ciation (NYHA) classifi cation system. After adjustments for age, gender, and BMI, serum A-FABP concentrations in patients with CHF were signifi cantly higher than in non-CHF patients (1.17 [6.63-19.93] ng/ml vs. 5.67 [3.20-8.87] ng/ml, P < 0.001), and signifi cantly progressed with the NYHA class (5.67 [3.20-8.87] ng/ml vs. 9.93 [5.91-15.61] ng/ml vs. 11.26 [6.64-21.55] ng/ml

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A702 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES

2834-PO 2836-PO Strain Ddifferences in the Stimulation of Incretin Hormones by Lipid Feeding ALISON B. KOHAN, STEPHANIE M. YODER, LENA OHLSSON, QING YANG, BO AHRÉN, PATRICK TSO, Cincinnati, OH, Indianapolis, IN, Lund, Sweden WITHDRAWN The incretin hormones, glucagon-like peptide-1 (GLP-1) and glucose-depen- dent insulinotropic polypeptide (GIP), are secreted by enteroendocrine cells in the gut following meal ingestion and enhance insulin secretion. Incretin hormones are released after the ingestion of all types of macronutrients, but the regulation of their secretion has not been well established in vivo. This is partly due to the problem in analyzing the hormones in blood because circulating incretin hormones are rapidly digested by dipeptidyl-peptidase-4 (DPP-4). To allow studies on regulation of incretin hormone secretion in vivo, the conscious lymph fi stula mouse model has been developed. This model allows measurement in real time of lymphatic incretin hormone secretion (since lymph lacks DPP-4). We have used this model to measure incretin hor- mone secretion after a mixed meal in three different mouse strains; we were interested in assessing whether there are underlying differences in incretin secretion between the most commonly used mouse strains. C57BL/6, 129/ SvJ, and FVB/N mice received a single intraduodenal bolus of Ensure, after which lymph was continuously collected for 3 h and analyzed for TG, glu- cose, GLP-1, and GIP. We found that in response to the Ensure bolus, incretin hormone secretion was clearly stimulated in all 3 strains of mice, although the time of peak hormone secretion was different between strains. Addi- tionally, there are signifi cant differences between strains in the pattern of TG and glucose secretion into lymph. We speculate that different strains of mice may display unique incretin hormone secretion patterns when chal- lenged with a meal, most likely because the mechanisms by which GIP and GLP-1 secretion is stimulated by lipid ingestion are different between inbred mouse strains. Furthermore, since many labs use different strains of mice for generating knockouts, our results are important for identifying different metabolic outcomes in animals with different genetic backgrounds. Supported by: DK-076928, DK-059630, F32-091173-01 2837-PO Possible Involvement of Bilirubin in Diabetic Vascular Infl amma- 2835-PO tion, Detected by its Specifi c Marker Pentraxin 3 Regulation of Specifi c Intestinal Cells Functional Activity by the YUICHI TAKASHI, TAKASHI WATANABE, YOKO MATSUZAWA, JUN SAITO, Glucagon-Like 1 Peptide Receptor MASAO OMURA, TETSUO NISHIKAWA, Yokohama, Japan, Tokyo, Japan MAMDOUH H. KEDEES, MARINE GRIGORYAN, YELENA GUZ, GLADYS TEITEL- Chronic infl ammation of blood vessels is known to promote the progres- MAN, Brooklyn, NY sion of atherosclerosis in diabetic patients, but no infl ammation marker highly We studied the localization and functional activity of the glucagon-like specifi c to blood vessels has been established yet. In this study, we investi- peptide-1 receptor (Glp-1r) in the gut. CD-1 mice were used to examine GLP- gated the usefulness of Pentraxin 3 (PTX3) as a specifi c marker of vascular 1r expression in ileum and the effects of its inhibition at the cellular and infl ammation. Blood and urine samples were obtained under written informed molecular levels. In ileum, Glp-1r mRNA was two fold higher in epithelial consent from 53 inpatients of our Center, and 35 of them had type 2 diabetes. cells than non-epithelial tissues (N=3 mice/experiment). Western blot analy- PTX3, high-sensitive CRP (hs-CRP), and 8-epi-PGF2_ level were measured, sis of isolated mucosal cell detected a major 43kDa band that is also found and their relationships with various clinical parameters were examined. in pancreatic islets. By immunohistochemistry, the receptor was localized to PTX3 level was not signifi cantly correlated with the hs-CRP, 8-epi-PGF2_ or Paneth but not to proliferating mucosal cells of the transit amplifying (TA) HbA1c level. It was signifi cantly positively correlated with HbA1c, T-Bil, and region, goblet or L cells. Immunostaining was eliminated by pre-incubation negatively correlated with LDL, TG, and T-cho. In the diabetic patients, the of the antibody with a specifi c blocking peptide. The GLP-1r was expressed PTX3 level was signifi cantly correlated with the T-Bil level but not with other by GLP-1 and Ki67 negative cells in the base of the crypt of duodenum, je- parameters. On multiple regression analysis of HbA1c, T-Bil and T-chol, the junum and ileum, but not of colon. The GLP-1r positive cells were PAS posi- T-Bil level showed the highest correlation among the 3 variables although tive Paneth cells. To test whether the Glp-1r regulates Paneth cell activity, the differences were not signifi cant. PTX3 belongs to the PTX superfamily we performed microarray analysis of ileal mucosal cells of mice with global along with CRP. CRP is secreted by the liver under stimulation by IL-6 and deletion of the glucagon receptor (Gcgr-/-) characterized by elevated levels is not highly specifi c to vascular infl ammation. PTX3, on the other hand, is of circulating Glp-1, littermate controls and Gcgr-/- mice injected with the expressed in vascular endothelial cells of patients with arteriosclerosis and Glp-1r antagonist exendin 9-39. Gcgr-/- mice showed signifi cant increase in is considered to be more specifi c to the blood vessels. In this study, the PTX3 the markers of Paneth cell activation Angiogenin 4 (Ang4) and Cd24 antigen level showed no correlation with the hs-CRP or 8-epi-PGF2_ level, suggesting and in Wntless homolog (Wls), a marker of differentiation. The increased that it refl ects different aspect of chronic vascular infl ammation. T-Bil showed expression of these molecules was downregulated by exendin 9-39. Glp-1r a positive correlation with the PTX3 level. PTX3 has been reported to have protein was also localized to PGP 9.5 positive enteric neurons of the my- a vessel protecting effect, and T-Bil is also considered to be secreted as an

enteric plexus. In ileum segments in vitro, the GLP-1r agonist Exendin-4 antioxidant against chronic infl ammation. Thus the measurement of T-Bil may Obesity (100nM, 30 min) induced mRNA levels of the early response gene c-fos. This contribute to the estimation of chronic vascular infl ammation refl ected by the upregulation was blocked by pre-incubation with a voltage-gated sodium PTX3 level, although further studies are required. PUBLISHED ONLY

channel blocker tetrodotoxin (TTX, 1μM). These fi ndings suggest that GLP- Integrated Physiology/ 1 participates in the regulation of Paneth and progenitor cells activity and 2838-PO myenteric neuron function Both Nateglinide and Acarbose Improve Postprandial Ghrelin Re- Supported by: Merck sponse in Patients With Newly Diagnosed Type 2 Diabetes FENPING ZHENG, XUEYAO YIN, WEINA LU, JIAQIANG ZHOU, HONG LI, Hangzhou, China Abnormal ghrelin regulation is linked to obesity-associated conditions, including type 2 diabetes (T2DM). We compared ghrelin response in newly- diagnosed T2DM patients and BMI-matched normoglycemic controls, and determined the effect of two postprandial hypoglycemic agents on ghre- lin response in T2DM. Open-label, randomized, prospective study in forty

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A703 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES

newly-diagnosed T2DM patients, randomly allocated to receive 120 mg performed. To evaluate the overall estrogenic activity, using the e-screen Nateglinide or 50 mg Acarbose three times a day for 4 weeks, and twenty proliferation cellular bioassay, of the serum of girls and adolescents with BMI-matched normoglycemic controls. Daily mean average glucose excur- T1D compared to healthy controls. We studied prepubertal girls with T1D sion (MAGE) was assessed using a continuous glucose monitoring system (T1Dg, N=8) and adolescents with T1D (T1Da, N=10), who had a gynecologi- and short-term sustained hyperglycemic state was evaluated via serum cal age greater than 2 years and were younger than 20 yr. These patients glycated albumin (GA). Fasting, 30 min and 120 min postprandial serum were compared with healthy prepubertal girls (N=9) and healthy adoles- ghrelin levels were determined by ELISA. Absolute postprandial ghrelin sup- cents (N=12). The overall serum estrogenic activity was evaluated by the pressions at 30 min and 120 min and ghrelin area under the curve (AUCGH) e-screen bioassay. This assay evaluates proliferation of MCF-7 BUS cells, were calculated. T2DM patients had similar fasting ghrelin levels (P = 0.546) which are known to be estrogen sensitive. Proliferation was assessed by and reduced postprandial ghrelin suppressions at 30 min and 120 min (P < fl uorometry (CyQuant® kit). Results were expressed as the relative response 0.01) compared to BMI-matched normoglycemic controls. Both Nateglinide observed to a serum pool obtained from healthy women. E-screen bioassay and Acarbose increased postprandial ghrelin suppression at 120 min and showed good correlation with the serum levels of 17`-estradiol (r = 0.87, p reduced AUCGH compared with baseline (P < 0.05), while only Nateglinide = 0.0002). T1Da exhibited lower estrogenic activity (55 ± 13) compared with increased postprandial ghrelin suppression at 30 min (P <0.05), which was controls (73 ± 27) (p < 0.05, Mann-Whitney test). Prepubertal girls exhibited positively correlated with the change of the early-phase insulin secretion similar serum estrogenic activity in both groups. This is the fi rst report show- by Nateglinide therapy. In stepwise multiple regression analysis, GA was ing lower serum estrogenic activity in T1D adolescents. These preliminary independently correlated with AUCGH (` = 0.46, P = 0.004) and HOMA-IR (` fi ndings suggest that T1D affects estrogenic activity, which may lead to the = 0.43, P = 0.009), and MAGE was independently correlated with HOMA-IR loss of physiologic estrogen protection against cardiovascular disease in (r = 0.42, P = 0.007) and ghrelin suppression at 30 min (` = -0.31, P = 0.042) premenopausal women with T1D. after 4 weeks of Nateglinide and Acarbose therapy. T2DM patients have Supported by: Fondecyt 1100123 reduced postprandial ghrelin suppression. Improved ghrelin regulation most likely plays an important role in glucose homeostasis in T2DM patients with 2842-PO Natelinide or Acarbose therapy. Liraglutide is More Effective Therapy than Insulin in Well-Con- trolled Japanese Patients With Type 2 Diabetic Mellitus 2839-PO HIROAKI SATOH, SATORU YAMAZAKI, YOSHIYUKI SUGAYA, NORITAKA MACHII, HIROYUKI HIRAI, KOJI HASEGAWA, SHINICHI NAKAJIMA, TSUYOSHI WA- TANABE, Fukushima, Japan WITHDRAWN The Action to Control Cardiovascular Risk in Diabetes (ACCORED) trial raised the possibility that hypoglycemia and weight gain may be associated with the increase in the risk of the cardiovascular mortality. Improved glyce- mic control with insulin therapy is associated with increase in body weight and hypoglycemia. In contrast to insulin, liraglutide is a once-daily human glu- 2840-PO cagon-like peptide-1 (GLP-1) receptor agonist that improves overall glycemic Analysis of Serum HMGB1 Levels in Japanese Obese Patients With control with weight loss and low risk of hypoglycemia. We therefore inves- Type 2 Diabetes tigated whether liraglutide reduces hyperglycemia safely without a marked AYAKO ISHIBASHI, MARI INOUE, SEIKI HIRANO, HIROSHI TAKATA, YOSHITAKA fl uctuation toward hypoglycemia compared with insulin therapy. Inclusion KUMON, YOSHIO TERADA, SHIMPEI FUJIMOTO, Nankoku, Japan criteria were controlled HbA1c < 6.9% with insulin therapy. Twelve sub- The high mobility group box-1 protein (HMGB1) is a ubiquitous nuclear jects with type 2 diabetes were enrolled; 75% male, (mean ± SD) age 64.2 ± 2 protein that determines nucleosomal stability binding transcription factors. 13.2 year, body weight 69.0 ± 12.9 kg, BMI 27.6 ± 12.9 kg/m , HbA1c 6.4 ± Serum HMGB1 levels may be derived from release from necrotic cells and 0.4%, dose of insulin 14.1 ± 6.6 U/day (0.21 ± 0.12 U/kg). These insulin treated activated immune cells. HMGB1 is highly expressed in endothelial cells, subjects were switched to liraglutide. There was no change in other medica- vascular smooth muscle cells and macrophages in atherosclerotic lesions. tions. Liraglutide was given once-daily with stepwise dose increments of 0.3 Taken together, HMGB1 is a putative marker of atherosclerosis. Chronic mg/day to 0.9 mg/day. After 3 months, switching from insulin to liraglutide microinfl ammation is involved in the pathogenesis of diabetic vascular signifi cantly improved HbA1c (-0.3%; p<0.05) and reduced body weight (-2.8 complications. However, little is known of the association of serum HMGB1 kg; p<0.05). In addition, during liraglutide treatment, average fasting blood levels with clinical parameters in patients with type 2 diabetes (T2DM). We glucose (FBG) levels for 30 days measured by the self-monitored BG (SMBG) recruited 112 Japanese patients with T2DM [79 men and 33 women, age: data were signifi cantly decreased from 137.2 mg/dl to 107.7 mg/dl (p<0.01). 64 ± 12 years, BMI: 24.8 ± 4.0 (mean ± SD)] treated with or without piogli- Furthermore, mean coeffi cient of variation (CV) for the within-person varia- tazone (PIO) including 54 patients with obesity (BMI*25) [Obese group (OG)] tion in self-measured FBG was lower with liraglutide treatment than with and 58 without obesity (BMI<25) [non-obese group (NOG)]. Serum HMGB1 insulin treatment, (9.1 vs. 28.6%, difference: 19.5%, p<0.01). In conclusion, concentrations were measured using an enzyme-linked immunosorbent as- liraglutide treatment is safe, well tolerated, and further improved glycemic say (SHINO-TEST, Japan). In OG, serum HMGB1 levels were correlated with control with low risk of hypoglycemia and sustained weight loss in subjects high-sensitive CRP (r=0.321, P<0.02) and adiponectin concentrations (r=- reaching target with insulin treatment. 0.279, P<0.05), but they were not in NOG. HMGB1 levels were not correlated with fasting plasma glucose or A1c in either group. Adiponectin concentra- 2843-PO tions were higher in patients treated with PIO than those in patients not Expression of the Glucagon-Like Peptide-1 Receptor in Psoriasis treated with PIO in both groups. Interestingly, HMGB1 levels were lower in Plaques patients treated with PIO (n=24) than those in patients not treated with PIO ANNESOFIE FAURSCHOU, JENS PEDERSEN, METTE GYLDENLØVE, STEN S. (n=30) (0.83 ± 0.36 vs. 1.66 ± 1.54 ng/ml, respectively, P<0.02) in OG, while POULSEN, JENS J. HOLST, CLAUS ZACHARIAE, LONE SKOV, TINA VILSBØLL, PIO did not affect HMGB1 levels in NOG. Thus, serum HMGB1 levels, a puta- FILIP K. KNOP, Hellerup, Denmark, Copenhagen, Denmark

Obesity tive marker of atherosclerosis, are associated with microinfl ammation that Psoriasis vulgaris is a chronic infl ammatory skin disease affecting 2-3% of might be reduced by pioglitazone in obese T2DM patients. the Western population. Recent case reports suggest that treatment with glucagon-like peptide-1 (GLP-1) receptor agonists may result in clinical im- PUBLISHED ONLY Integrated Physiology/ 2841-PO provement of the disease. The aim of the present study was therefore to Estrogenic Bio-Activity in Girls and Adolescents With Type 1 Dia- investigate if GLP-1 receptors (GLP-1Rs) are present in human skin and if betes expression of the GLP1R is up-regulated in patients with psoriasis. Two 3 DANIELA MARTÍNEZ, M. CECILIA LARDONE, ANDREA CASTRO, GERMAN mm-punch skin biopsies were taken from 6 healthy volunteers (one biopsy ® IÑIGUEZ, PATRICIA LÓPEZ, PAULINA M MERINO, FERNANDO CASSORLA, ETHEL was immediately submerged in RNAlater and one was transferred to 4% CODNER, Santiago, Chile paraformaldehyde). From 6 psoriasis patients, four 3 mm-punch skin biopsies Premenopausal women with type 1 diabetes (T1D) have an increased risk were taken; two from unaffected skin and two from psoriasis plaques. Biop- of cardiovascular disease mortality compared with healthy women of similar sies from unaffected and affected skin were either immediately submerged ® age. These data suggest that the physiologic estrogen protection conferred in RNA-later or transferred to 4% paraformaldehyde. The fi xed biopsies against cardiovascular disease is lost in these patients. However, studies were paraffi n-embedded and examined for GLP-1R presence using immuno- evaluating serum estrogenic activity in patients with T1D have not been histochemical staining. Total RNA was extracted, reverse transcribed and

For author disclosure information, see page 797. ADA-Funded Research

A704 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES analyzed for the expression of GLP1R using SYBR® Green-based qPCR. Suf- plasma adiponectin was increased 3-fold in PIO and PIO+ALO compared with fi cient RNA was isolated from biopsies and integrity was confi rmed by gel CON (CON= 3.6 ± 0.2, ALO= 3.7 ± 0.1, PIO= 10.5 ± 0.7, PIO+ALO= 10.2 ± 0.5 electrophoresis before it was reverse transcribed to cDNA. Gene expres- μg/ml; P<0.001). Thus, pioglitazone alone and in combination with alogliptin sion analysis showed expression of GLP1R in 5 out of 6 skin biopsies (83%) delays diabetes onset in the UCD-T2DM rat. Pioglitazone and alogliptin ap- from psoriasis plaques whereas GLP1R expression was found in signifi cantly pear to act additively to improve glucose metabolism. (P=0.02) less biopsies from unaffected skin (2 out of 12 (8.3%) - 1 from a pso- Supported by: Takeda Pharmaceuticals North America, Inc. riasis patient and 1 from a healthy volunteer). Our results provide evidence for expression of GLP1R in psoriasis plaques, which may contribute to under- 2846-PO stand the positive effect of GLP-1R agonist treatment on psoriasis described Effect of Acute Hyperinsulinemia and Angiotensin II Type 1 Recep- in recent case reports. Thus, a direct effect of GLP-1 receptor activation on tor Blockade (ARB) on Androstane and Pregnane Metabolites in psoriasis can not be excluded. Healthy Men and Men With Type 2 Diabetes KATERINA ZIDKOVA, MARTIN HILL, EVA SVEHLIKOVA, JANA VRBIKOVA, TEREZIE 2844-PO PELIKANOVA, Prague, Czech Republic Role of GLP-1 and Insulin in the Regulation of ƴ Cells in Humans The association between lower testosterone levels and type 2 diabe- GIOVANNA MUSCOGIURI, TERESA MEZZA, GIAN PIO SORICE, ANNAMARIA tes in men was delineated; however the responsible mechanisms were PRIOLETTA, LORELLA MARSELLI, JENS I. HOLST, PIERO MARCHETTI, ANDREA not identifi ed till now. We focused on the effect of acute hyperinsulinemia GIACCARI, Rome, Italy, Pisa, Italy, Copenaghen, Denmark and ARB on the plasma levels of androstanes and pregnanes in men with In patients with type 2 diabetes fasting hyperglucagonemia as well as type 2 diabetes (D) and healthy controls (H). Eleven D and 12 age-matched increased glucagon response to meal ingestion contribute to hyperglycemia. H underwent hyperinsulinemic (1mU/kg/min) euglycemic (5 mmol/L) clamp The hormonal mechanism that regulates glucagon secretion is not well elu- (HEC) and HEC after ARB (losartan 200 mg, AT-HEC). Serum concentrations cidated. GLP 1 has been shown to have a powerful ability to lower glucagon of selected steroids were measured using GC-MS before and at 240 minutes secretion although it is still debated if GLP 1-induced inhibition of glucagon of HEC and AT-HEC. Enzymatic activities were assessed as product/precur- secretion is a direct effect of GLP 1 on _ cell or it is mediated by paracrine sor ratios. Repeated measures ANOVA, followed by least signifi cant differ- effects of somatostatin or insulin elicited by GLP 1. To address this question ence multiple comparisons, were used for data evaluation. D had decreased we studied 10 patients (7F/3M) before undergoing to partial pancreatectomy insulin sensitivity, assessed by metabolic clearance rate of glucose (MCR) with a 240-min meal test assaying plasma concentration of GLP-1, gluca- (p<0.001), which was not affected by ARB in either group. T(p<0.001) and gon, insulin, C-peptide and glycaemia; Hyperinsulinemic Euglycemic Clamp DHT (p<0.001) levels, as well as 5_-reductase activity (DHT/T, A3a5a/A2 ra- was performed to evaluate insulin sensitivity. During surgery we collected tios) (p<0.001) were lower in D compared to H. We found consistently higher pancreas specimens in 7 patients on which we performed morphometry and A3a5aC/A3b5aC, Preg20aC/PregC, AdiolC/DHEAC and Adiol/DHEA, A3a5b/ staining for glucagon. The mean age of patients was 61±19.3 years, with BMI A2, A3a5b/A3a5a ratios in D. Hyperinsulinemia affected some of these ra- 27.9±5.3 kg/m2. Insulinsensitivity was related to the area under the curve of tios identically in D and H. There were no differences in androstanes and glucagon secretion during meal test (r=-0.79;p=0.030) and to the percent- pregnanes during HEC compared to AT-HEC in both groups. We conclude that age of areas positive for glucagon in relation to the total pancreatic tissue men with type 2 diabetes have changes in steroid metabolism with consis- (GlucA/TA (r=-0.88; p=0.008). Interestingly, insulin and glucagon secretion tently increased activities of enzymes from aldoketo-reductase family 1. We were inversely related (r= -0.63; p=0.047) and GlucA/TA (r=-0.76;p=0.046). did not prove any signifi cant effect of acute ARB. Although GLP 1 secretion was related to insulin response (r=0.67; p=0.033), it did not correlate with glucagon (r=-0.42; p=NS). These results support the hypothesis that the effect of GLP 1 on _ cells is mostly mediated by insulin secretion, but whether this is due to a paracrin effect or to circulating insulin cannot be deduced. . Furthermore, insulin levels may play a role in determin- ing the morphology of a cells, as shown by the tight relationship of insulin secretion with GlucA/TA. The relation between glucagon positive area and secretion with insulin resistance open new perspectives in the understand- ing of the regulation of the secretion of these hormones.

2845-PO Chronic Administration of Pioglitazone Alone and in Combination With Alogliptin Delays the Onset of Type 2 Diabetes in UCD-T2DM Rats BETHANY P. CUMMINGS, JAMES L. GRAHAM, KIMBER L. STANHOPE, PETER J. HAVEL, Davis, CA Due to the increasing prevalence of type 2 diabetes (T2DM), there is a need to identify strategies for diabetes prevention. We have investigated the effi cacy of pioglitazone, a PPAR agonist, and alogliptin, a DPP-IV inhibi- Supported by: MZO 00023001 tor, alone and in combination to prevent T2DM onset in UCD-T2DM rats, a model of polygenic obese T2DM. At 2 mo of age, rats were divided into three 2847-PO groups: Control (CON), Pioglitazone (PIO; 2.5 mg/kg/d), Alogliptin (ALO; 20 mg/kg/d) and Pioglitazone and Alogliptin in combination (PIO+ALO) (n=16). Rats received the drug in ground chow. Non-fasting blood glucose was mea- sured weekly to determine diabetes onset (>200 mg/dl) and an OGTT was WITHDRAWN

conducted at 3.5 months of age. Energy intake and body weight at 3 mo Obesity were higher in PIO and PIO+ALO compared with CON and ALO (CON= 32 ± 1 kcal/d, 531 ± 5 g; ALO= 32 ± 1 kcal/d, 530 ± 7 g; PIO= 34 ± 1 kcal/d, 560 ± 6 g; PUBLISHED ONLY PIO+ALO= 35 ± 1 kcal/d, 563 ± 7 g; P<0.001). The number of diabetes free days Integrated Physiology/ was greater in PIO and PIO+ALO groups (CON= 170 ± 15, ALO= 169 ± 13, PIO= 239 ± 3 and PIO+ALO= 236 ± 3 days; P<0.001). T2DM incidence was lower in PIO and PIO+ALO groups (CON= 13/16, ALO= 14/16, PIO= 1/16 and PIO+ALO= 2/16). Fasting plasma glucose and insulin were lower in PIO and PIO+ALO groups compared with CON. Fasting plasma FFA were reduced in all treat- ment groups compared with CON at multiple time points (P<0.05). During the OGTT, the glucose AUC was lower in PIO, ALO and PIO+ALO compared with CON (CON= 10929 ± 913, ALO= 8772 ± 957, PIO= 7371 ± 824, PIO+ALO= 6269 ± 540 mg/dl × 120 min; P<0.05) and the glucose AUC was lower in PIO+ALO compared with ALO and PIO (P<0.05). Starting at 3 months of age, fasting

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A705 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES

2850-PO

WITHDRAWN 2848-PO Does Bile Acid Signaling via FGF-19 Contribute to the Early Thera- peutic Effects of Roux-en-Y Gastric Bypass Surgery? ELIZABETH O. BEALE, DENISE HASSON, NAMIR KATKHOUDA, PETER CROOKES, 2851-PO MIN LIN, DAVID SMITH, BARRY M. FORMAN, Los Angeles, CA, Duarte, CA Roux-en-Y gastric bypass (RYGB) leads to rapid improvement in glucose tolerance that is greater than that after gastric banding (GB). An understand- ing of the mechanism of benefi t of RYGB may lead to novel pharmacologic alternatives. Alterations in bile acid (BA) signaling may contribute to this WITHDRAWN benefi t as: 1) circulating BAs have been reported to be signifi cantly high- er following RYGB and 2) BAs are now known to regulate energy balance through distinct pathways, including via induction of intestinal FGF15 (ani- mals)/FGF19 (humans). FGF19 administered to animals decreases adiposity and improves glucose tolerance. We hypothesize that circulating FGF19 will be elevated early post RYGB but not GB. In this pilot study we measured plasma FGF19 pre- and early (1-2 weeks) post-op in fasting, obese patients, of varying diabetes status, without cholecystectomy undergoing either GB or RYGB. See table. There was no signifi cant difference in age or BMI of subjects preoperatively. There was large inter-individual variation in FGF19 both pre- and post-op but the fold change was higher post-RYGB and there was a signifi cant increase in FGF19 following RYGB but not GB. Thus pilot data suggests that RYGB but not GB leads to early increase of FGF19 which may therefore be a key factor in the rapid increase in glucose tolerance seen post- RYGB. This study justifi es ongoing investigation into a role for FGF19 as a novel therapeutic agent for obesity and diabetes, including assessments at later time periods and in the fed state. GB vs. RYGB Operation N M:F Age (years) BMI Fold Paired 1-tailed [mean(sd)] Pre Op Change t-Test Log Pre Op (kg/m2) vs.Log Post Op [mean(sd)] (p-value) GB 7 3:4 48.6 (7.0) 48.6 (7.0) 1.5 (0.7) 0.19 RYGB 15 3:12 43.3 (13.3) 43.3 (13.3) 2.4(2.0) 0.01 2852-PO 2849-PO The Effect of Sitagliptin on Glycemic Control, Adiponectin and Lipid GLP-1 Levels are Markedly Increased 1 Month after Laparoscopic Metabolism Mini-Gastric Bypass Surgery in Non-obese Patients With Type 2 KENICHIO NISHIKAWA, TATSUO YANAGAWA, TAKATERU AKIYAMA, YUHKO Diabetes KIMURA, RIEKO MORINAGA, Tokyo, Japan HYEONGKYU PARK, SEOKCHUN YEOM, MYUNGJIN KIM, SUNGWAN JEON, DPP4 has reported to be one of adipocytokines. The aim of this study is KYOIL SUH, KYUNGYUL HUR, DONGWON BYUN, Seoul, Republic of Korea to evaluate the effects of sitagliptin on glycemic control, adiponectin, and Gastric bypass surgery(GBP) in morbidly obese patients often results in lipid metabolism in diabetic subjects. We included consecutive 40 patients weight loss, increase in incretin levels, and improvement of T2DM. In Asian with type 2 diabetes mellitus (80% males, 62 years of age, BMI 25.9 kg/ countries, T2DM incidence is recently increasing in non-obese as well as m2) who started to take 50mg sitagliptin once daily (the sitagliptin group) obese subjects. Mini-GBP involves creating a long-sleeved gastric tube along and consecutive 20 patients with type 2 diabetes mellitus (80% males, 62 the lesser curvature, and a Billroth II type loop gastroenterostomy with the years of age, BMI 25.1 Kg/m2) who did not take sitagliptin served as con- side of small bowel. Compared to Roux-en-Y GBP, mini-GBP is a simpler and trols (the control group). The levels of serum adiponectin, fasting plasma safer procedure, and therefore, the preferred GBP for patients with moder- glucose (FPG), glycosylated hemoglobin (HbA1c) and serum lipid parameters ate obesity. We investigated the effects of mini-GBP on incretin responses were measured at the study entry and 3 months after the entry. We did to oral glucose tolerance test(OGTT) in non-obese (body mass index[BMI] < not change the medication other than sitagliptin during the 3 months. The 30 kg/m2) patients with T2DM. A total of 12 patients (7 M/5 F, age 45 ± 3 yr, General Linear Model, including repeated measures, was used to analyze BMI 26.2 ± 0.6 kg/m2) who received laparoscopic mini-GBP were included. the results. The value for HbA1c is estimated as an NGSP equivalent value Active glucagon-like peptide-1(GLP-1), gastric inhibitory peptide(GIP), insulin, calculated by the formula HbA1c = HbA1c (JDS) + 0.4. There was a signifi cant and c-peptide levels were measured during 75-g OGTT before and 1 month reduction in the sitagliptin group compared with the control group regarding after mini-GBP. One month after surgery, body weight decreased by 6.2 ± 0.7 HbA1c (JDS) (7.35% to 6.50% vs. 6.21% to 6.30%, P < 0.01), FPG (170 mg/dl

Obesity kg, postprandial(pp) glucose levels signifi cantly decreased(pp 90 min: 335 ± to 142 mg/dl vs.140 mg/dl to 159 mg/dl, P < 0.01) and serum TG (188 mg/dl 11 vs. 269 ± 25 mg/dL, P < 0.05; pp 120 min: 317 ± 17 vs. 232 ± 30 mg/dL, P < to 157 mg/dl vs. 136 mg/dl to 154 mg/dl, P < 0.05). In addition, there was a 0.05). Oral glucose-stimulated insulin and c-peptide levels increased, but not signifi cant increase in the sitagliptin group compared with the control group PUBLISHED ONLY Integrated Physiology/ signifi cantly(insulin area under curve[AUC]: 36 ± 10 vs. 57 ± 14 uIU/mL·min-1; regarding serum adiponectin (8.7 ug/ml to 10.6 ug/ml vs. 10.3 ug/ml to 9.6 c-peptide AUC: 6.8 ± 0.9 vs. 9.1 ± 1.5 ng/mL·min-1). Active GLP-1 levels after ug/ml, P < 0.01) and serum apolipoprotein A-1 (129 mg/dl to 134 mg/dl vs. 150 OGTT markedly increased 3-fold(peak: 207 ± 47 vs. 619 ± 133 pg/mL, P < mg/dl to 147 mg/dl, P <0.05). We have demonstrated that in patients with 0.001; AUC: 108 ± 18 vs. 311 ± 56 pg/mL·min-1, P < 0.001). However, total GIP type 2 diabetes mellitus sitagliptin not only improves the glycemic control levels signifi cantly decreased after mini-GBP(peak: 238 ± 28 vs. 137 ± 16 pg/ but also increases serum adiponectin and apolipoprotein A-1 and decreases mL, P < 0.001; AUC: 184 ± 20 vs. 98 ± 10 pg/mL·min-1, P < 0.001). In summary, serum triglycerides. These results may suggest sitagliptin is most likely to GLP-1 levels were markedly increased after mini-GBP in non-obese patients have anti-atherosclerotic effects. with T2DM. Further studies will be needed to address the long-term effects of mini-GBP on glucose homeostasis in non-obese patients with T2DM.

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A706 INTEGRATED PHYSIOLOGY—OTHERCATEGORY HORMONES

2853-PO groups in the early phase of OGTT and then decreased signifi cantly at 180 min in healthy control group (P<0.05). Our study suggested the different se- cretion pattern of GLP-1 and GIP in PCOS obese and non-obese patients may WITHDRAWN contribute to the different levels of C-peptide and LH and thus be involved into the multiply pathogenesis of PCOS. Incretin may not only stimulate the secretion of insulin but also play a role on the central nervous system. Supported by: National Science Foundation of China (30971398)

2854-PO 2856-PO Association of Increased Angiotensin II and Decreased Osteocalcin Are Prediabetics With an Elevated CRP More Likely to Progress to in Diabetes Diabetes? RAELENE E. MASER, M. JAMES LENHARD, PAUL KOLM, Newark, DE SHASHI K. AGARWAL, East Orange, NJ Recent studies in animal models and humans have suggested that the High sensitivity C-reactive protein (CRP), a marker of infl ammation, is asso- skeleton exerts an endocrine regulation of glucose metabolism through an ciated with diabetes and cardiovascular disease. However, studies examin- osteoblast-specifi c hormone, osteocalcin (OC). Several variables (e.g., BMI, ing the association between CRP and prediabetes among hypertensives with increased leptin, chronic hyperglycemia) have been shown to decrease OC and without diabetes are limited. This study was done to determine whether levels. Furthermore, angiotensin II (ang II) has been shown to inhibit the ex- CRP measurement in hypertensive prediabetics and diabetics has any clini- pression of mRNA for OC in osteoblastic cells in rodent models. Whether cal signifi cance. A retrospective chart review was done on 200 consecutive ang II contributes to decreased OC levels in individuals with diabetes has hypertensive patients who had both HbA1c and CRP recorded. HbA1c was not been explored. We investigated the association between ang II and OC categorized as follows: normal: <5.7(less than 39 mmol/mol); pre-diabetes: levels in 57 individuals with diabetes (53% type 1, 47% type 2). Ang II, af- 5.7% - 6.4% (39 - 46 mmol/mol); diabetes: 6.5% (47 mmol/mol) or higher. CRP ter collection with protease inhibitors, was measured via RIA and intact OC was categorized as follows: normal: 0.9mg/L or less; high: 1mg/L or more. levels were measured by ELISA. Participants (56% men) were aged 51±12 Of the 200 patients, there were 84 males (M) and 116 females (F). Of these, (mean±SD) yrs, duration of diabetes 16±12 yrs, HbA1c 7.6±1.1%, and 67% 141 had CRP 0.9 mg/L or less (M90; F51); Of these 141 with normal CRP lev- were using ACE-inhibitors/ARBs. OC was negatively correlated with ang II els, HbA1c status was as follows: normal: 38 (27%) (M27; F12); prediabetes: (r = -0.36, p<0.01), BMI (r = -0.45, p<0.001), age (r = -0.48, p<0.001), type of 71(50%) (M47; F24); diabetes 32(23%) (M19; F13). 59 had CRP > 1mg/L (M33; diabetes (r = -0.38, p<0.01), and ACE-inhibitors/ARBs use (r = -0.38, p<0.01). F26); Of these 59 with elevated CRP, HbA1c status was as follows: normal Ang II levels were similar for those on ACE-inhibitors/ARBs vs. not (31±13 15 (26%) (M7; F8), prediabetes 22 (37%) (M12; F10); diabetes 22 (37%) (M5; vs. 28±9 pg/mL, p=0.4) whereas OC was lower for those on ACE-inhibitors/ F17). Of the 53 with normal glucose metabolism, 39 (72%) had normal CRP ARBs vs. not (7±4 vs.10±4 ng/mL, p<0.01). Table 1 shows the results of lin- and 15 (28%) had high CRP; Of the 93 with prediabetes, 71 (76%) had normal ear regression with OC levels as the dependent variable (model R2=0.518, CRP and 22(24%) had high CRP; Of the 54 with diabetes, 32 (59%) had normal p<0.001). Ang II, BMI and glycemic control were independent correlates of CRP and 22 (41%) high CRP. This study showed that most hypertensive pre- OC. Ang II appears to have a physiologic function in the suppression of OC diabetics have normal CRP and this is comparable with those with normal in diabetes and may therefore affect OC’s role in the regulation of glucose glucose homeostasis. More diabetics have an elevated CRP when compared metabolism. to prediabetics. This suggests that progression from prediabetes to diabetes Table 1. Regression Analyses with Osteocalcin as Dependent Variable in hypertensives may be accompanied by a rise in CRP. Independent Variables Regression Coeffi cient (SE) p-value Angiotensin II (pg/mL) -0.117 (0.037) 0.003 2857-PO 2 Decreased Caveolin-1 Expression of Peripheral Blood Mononuclear BMI (kg/m ) -0.243 (0.079) 0.003 Cells in Newly Diagnosed Type 2 Diabetic Patients HbA1c (%) -0.897 (0.388) 0.025 WEI JIANG, SHUO LIN, PANWEI MU, XIAOFENG LI, LONGYI ZENG, Guangzhou, ACE-inhibitors/ARBs -1.922 (0.988) 0.058 China Age (yrs) -0.081 (0.044) 0.071 Caveolin-1, a positive mediator of insulin signaling, has recently been shown decreased expression of white adipose tissue in obese people and Type of diabetes 1.057 (1.156) 0.365 rats. However, its expression of circulating peripheral blood mononuclear Gender 0.163 (0.842) 0.847 cells (PBMCs) in newly diagnosed type 2 diabetic patients has not been re- Supported by: Delaware INBRE, NIH Grant 2 P20 RR016472-11 from the NCRR ported. To investigate it, we isolated PBMCs from 47 newly diagnosed type 2 diabetic patients (aged 46.12±10.74 years) and 41 healthy subjects (aged 42.34±10.88 years) by fi coll density gradient centrifugation. Insulin sensitiv- 2855-PO ity was assessed by homeostatic model assessment of insulin resistance Incretin Levels in Obese and Non-Obese Patients With Polycystic (HOMA-IR). Real time PCR and Western blotting analysis demonstrated that Ovary Syndrome caveolin-1 mRNA and protein levels of PBMCs were downregulated in newly YU LIU, LU WANG, Changchun, China diagnosed Type 2 diabetic patients compared with healthy subjects (P<0.05). Polycystic ovary syndrome (PCOS) is an endocrine disease and the patho- Furthermore, caveolin-1 mRNA expression of PBMCs was inversely corre- genesis of PCOS remains unclear. Some researches reported decreasing lated with BMI (r=-0.216, P<0.05), fasting plasma glouse (r=-0.443, P<0.01), levels of postprandial incretions which show an important factor of impaired glycated hemoglobin A1C (r=-0.579, P<0.01), HOMA-IR (r=-0.385, P<0.01) and insulin secretion in Type 2 Diabete(T2D) patients although the level of GLP-1 plasma C-reactive protein (r=-0.302, P<0.01). In conclusion, caveolin-1 expres- and GIP in PCOS patients and the relationship with beta call function still are sion of PBMCs is decreased in newly diagnosed type 2 diabetic patients which not fully elucidated.17 female patients diagnosed with PCOS were divided may contribute to worse glycemic control and lower insulin sensitivity. into obese group(n=9, BMI 25kg/m2 )and non-obese group(n=8, BMI<25kg/ * Supported by: National Natural Science Foundation of China m2 ). 6 female T2D patients (BMI 26.31±1.27 kg/m2) and 8 healthy women(BMI Obesity 21.43±1.53 kg/m2) were used as the control groups. All participants under- went 75 gram OGTT fro 3 hours. Blood samples were collected at 0, 30,60, 2858-PO PUBLISHED ONLY 120,180 minutes to detect the levels of glucose,insulin, C-peptide, GLP-1 and The Relation of Serum Neutrophil Elastase Concentration and Se- Integrated Physiology/ GIP. Serum sex hormones(PRL, E2, T, FSH, LH) were also analyzed in groups. rum Elastase Activity in Metabolic Syndrome The results show the area under curve (AUC) of C-peptide and the secret- BU KYUNG KIM, JAE HEE PARK, WOO MI KIM, YOUNG SIK CHOI, YO-HAN PARK, ing level at 60, 180 minutes in both PCOS obese and non-obese groups are Busan, Republic of Korea signifi cantly higher than in healthy control group(P<0.05). The HOMA-IR in Background: Human neutrophil elastase (NE) is one of the serine protease, PCOS obese patients are increased (P<0.01) while the ratio of LH/FSH are which plays an important role in potent antimicrbial activity and degradation decreased compared with non-obese patients (P<0.05). The level of GIP at of elastin. Elevation of NE level and activity has been demonstrated in a 180 min in obese PCOS patients are increased compared with that in healthy variety of pathological conditions including atherosclerosis, obesity, systolic controls (P<0.05). GLP-1 levels in obese PCOS group show a signifi cantly dif- hypertension and type2 diabetes mellitus. However, studies about NE in ferent time-dependent pattern (P<0.01 for obese PCOS versus time inter- metabolic syndrome (MS) are rare. The aim of this study is whether NE level action). GLP-1 concentrations were similar in non-obese PCOS and control and activity is related with metabolic components, whether it is elevated in

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A707 OBESITY—ANIMALCATEGORY

obese and hypertension, and whether it has a deleterious effect on pulmo- -0.33, P =0.006). In multiple regression analysis, we observed that the rela- nary function. Methods: A 138 health examinees who participated in health tionship between MMP-9 and adiponectin (`=-0.29; p=0.03) was indepen- screening at health promotion center in Kosin University, Gospel Hospital dent of BMI and insulin sensitivity. Our data suggests that anti-infl ammatory were enrolled in this study. They were measured the following, mass index, activity of adiponectin could have a negative infl uence on MMP-9 in young waist circumference, blood pressure, lipid profi le, high sensitivity C-reactive healthy men independently of BMI and insulin sensitivity. protein, serum NE level and activity, and pulmonary function tests including forced expiratory volume in one second (FEV1.0), forced vital capacity (FVC) 2861-PO and FEV1.0/FVC ratio. Results: There was positive correlations between NE activity and systolic hypertension. And also, NE activity was signifi cantly higher in stage 1 hypertension (0.0388 ± 0.01) than normotensive (0.0388 ± 0.01), prehypertensive (0.084 ± 0.09), and stage 2 hypertension (0.022 ± 0.00) (P<0.05, or p<0.01). Serum NE level was positively correlated with NE WITHDRAWN activity (r=0.172, p=0.04). Serum NE concentration was positively correlated with white blood cell (WBC) counts (r=0.223, p=0.009) and erythrocyte sedi- mentation rate (ESR) (r= 0.203, p=0.017) and NE activity was positively cor- related with ESR (r=0.231, p=0.007). FEV1 and FVC in MS were signifi cantly lower than non-MS. Conclusion: Serum NE activity was elevated in stage 1 hypertensive subjects and was positively correlated with systolic hyperten- sion. Serum NE level/activity was positive correlations with infl ammatory markers, such as WBC counts and ESR.

2859-PO

WITHDRAWN

OBESITY—ANIMAL

2862-PO A Short Term Repeated Administration of Low Dose Endocrine Dis- ruptors Associated With the Development of Diabetes In Adoles- cent Male Sprague-Dawley Rats MOON KI HONG, JAE WOONG KIM, SEONGHYUP HYUN, MI YI KIM, SEOK HONG LEE, JAETAEK KIM, SANG-YON KIM, YUN-JUNG YANG, YEON-PYO HONG, JI- HYUN AHN, Seoul, Republic of Korea Recently, the environmental obesogen hypothesis has been proposed, suggesting that environmental chemicals may contribute to the develop- ment of obesity and metabolic disorders such as diabetes. Bisphenol A (BPA) is one of the environmental chemicals that are widely used in daily life. BPA is present in drinking water, canned goods, and even milk bottles. This study was performed to investigate the association between the development of diabetes and environmental exposure level to BPA. Four week old OLETF rats were randomly assigned to three groups of fi ve animals and each group was given different concentration of corn oil with BPA (0, 0.001, 0.1 mg/kg/ day). Plasma insulin level was signifi cantly higher in rats exposed to BPA 0.1 mg/kg/day (P = 0.025) and 0.001 mg/kg/day (P = 0.018) than in control. Plasma malondealdehyde level was also higher in rats exposed to BPA 0.1 2860-PO mg/kg/day than in control (P = 0.017). The glucose levels were signifi cantly Adiponectin as a Negative Independent Predictor of MMP-9 (Matrix higher in the BPA 0.1 mg/kg/day treated group compared to controls at 90 Metalloproteinase-9) in Serum Concentration min after glucose administration (P = 0.008). These data indicate that the AGNIESZKA ADAMSKA, AGNIESZKA NIKOLAJUK, MONIKA KARCZEWSKA- impaired glucose tolerance was present in BPA treated OLETF rats. PPAR- KUPCZEWSKA, ELZBIETA OTZIOMEK, MARIA GORSKA, IRINA KOWALSKA, gamma protein expression levels in the whole cell lysate were decreased MAREK STRACZKOWSKI, Bialystok, Poland, Olsztyn, Poland in the BPA treated group compared to the vehicle-treated group. In conclu-

Obesity The imbalance between various infl ammatory biomarkers plays an impor- sion, low dose BPA exposure in adolescent OLETF rat may contribute to the tant role in the atherosclerosis process. Matrix metalloproteinase-9 (MMP- development of diabetes. 9) is one of the non-classical risk factors of cardiovascular diseases whereas PUBLISHED ONLY Integrated Physiology/ adiponectin has insulin-sensitizing, antiatherogenic and antiinfl ammatory 2863-PO properties. The aim of the present study was to analyze the associations be- The ATP-P2X7 Signaling Axis is Dispensable for Obesity-Associat- tween serum adiponectin and matrix metalloproteinase-9 concentrations in ed Infl ammasome Activation in Adipose Tissue subjects with normal glucose tolerance. We examined 96 young men (mean SHENGYI SUN, YEWEI JI, SHENG XIA, SANDER KERSTEN, LING QI, Ithaca, NY, BMI-25,2 kg x m-2) with normal glucose tolerance according to WHO crite- Wageningen, The Netherlands ria. Insulin sensitivity was measured with the hyperinsulinemic euglycemic Infl ammasome activation in adipose tissue has been implicated in obe- clamp technique. Serum adiponectin and MMP-9 concentrations were esti- sity-associated insulin resistance and type 2 diabetes. However, when and mated. Insulin sensitivity was positively related to adiponectin (r = 0.22, P= how infl ammasome is activated in adipose tissue remains speculative. Here 0.04). Serum adiponectin concentration was negatively associated with BMI we test the hypothesis that extracellular ATP, a potent stimulus of infl am- (r = -0.24, P =0.021) and positively correlated with HDL-cholesterol (r = 0.25, P masome in macrophages via purinergic receptor P2X, ligand-gated ion chan- =0.02). We found negative correlation between adiponectin and MMP-9 (r = nel, 7 (P2X7), may play a role in infl ammasome activation in adipose tissue

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A708 OBESITY—ANIMALCATEGORY in obesity. Our data show that infl ammasome is activated in adipose tissue plantation of this device in diet-induced obese (DIO) rats resulted in transient upon 8 week feeding of 60% HFD, coinciding with the onset of hyperglyce- body-weight reduction that was matched by pair feeding in sham animals. mia and hyperinsulinemia as well as the induction of P2X7. Unexpectedly, Consistent with these results, implantation of dELS was associated with a P2X7-defi cient animals on HFD exhibited no changes in metabolic pheno- signifi cant decrease in food intake during the fi rst 15 days post-op. Glucose types, nor in infl ammatory responses and infl ammasome activation compar- tolerance following a mixed meal challenge was also improved in dELS rats ing to the WT cohort. Similar observations were obtained in hematopoietic as compared to sham-control rats. To gain a greater understanding of dELS cell-specifi c P2X7-defi cient animals generated by bone marrow transplanta- implantation on glucose homeostasis, we repeated this initial study in the tion. Our results show that infl ammasome activation in adipose tissue in Zucker Diabetic Fatty-rat (ZDF), a rat model of impaired glucose handling. obesity coincides with the onset of hyperglycemia and hyperinsulinemia, Consistent with the effects observed in DIO rats, dELS implantation induced and unexpectedly, is not mediated by the ATP-P2X7 signaling axis. Thus, a signifi cant, but transient decrease in food intake and a sustained decrease the infl ammasome-activating danger signal(s) in adipose tissue in obesity in body weight throughout the study. The decreased body weight was asso- remain to be characterized. ciated with enhanced fat mass loss, while sparing lean mass, thus producing Supported by: HHMI, The Netherlands Nutrigenomics Center, Cornell Startup a more metabolically favorable body composition. dELS rats were also found Package, NIH to be more glucose tolerant than sham operated controls. Incresed glucose tolerance was associated with elevated GLP-1 and decreased insulin levels 2864-PO in response to a mixed-meal. Taken together, these results confi rm previous DDP-4 Inhibitor Prevents Neuronal Insulin Resistance, Brain Mito- reports suggesting metabolic improvements following dELS implantation. chondrial Dysfunction, and Impaired Learning and Memory Caused Furthermore, these studies suggest that these improvements may be as- by High-Fat Diet Consumption sociated with enhanced insulin sensitivity and GLP-1 secretion as well as SIRIPORN C. CHATTIPAKORN, NOPPAMAS PIPATPIBOON, HIRANYA PINTANA, remodeling of body composition. WASANA PRATCHAYASAKUL, NIPON CHATTIPAKORN, Chiang Mai, Thailand We previously demonstrated that 12-wk high fat diet (HFD) consumption 2866-PO could cause peripheral and neuronal insulin resistance, and brain mitochon- Infl uence of High Glucose Concentration on Bone Metabolism Pa- drial dysfunction in rats. Although vildagliptin is shown to improve periph- rameters in C57BL/6 Mice Fed With a High-Fat Diet eral insulin sensitivity, its effects on neuronal insulin resistance and brain RAUL H. BORTOLIN, SILVIA T. HIMELFARB, FABRICIO M. SANTOS, AGLAUCIELE mitochondrial dysfunction caused by HFD have never been investigated. We MARIA L. OLIVEIRA, FRANCISCO P. FREIRE-NETO, JOÃO F. BEZERRA, LEANDRO tested the hypothesis that vildagliptin prevents neuronal insulin resistance, VINICIUS F. MORAIS, MARCELA ABBOTT G. URURAHY, ANNE CAROLINE S. brain mitochondrial dysfunction and the impaired learning and memory BARBOSA, BARBARA S. LEMOS, FERNANDA A. BRITO, MARIA G. ALMEIDA, caused by HFD consumption. Male rats were divided into 2 groups to receive LUCIANA A. REZENDE, ROSARIO DOMINGUES C. HIRATA, MARIO H. HIARATA, either HFD or normal diet for 12 weeks. Then, each group was divided into ADRIANA A. REZENDE, Natal, Brazil, São Paulo, Brazil, Ribeirao Preto, Brazil two subgroups to receive either vildagliptin (3 mg/kg/day, p.o.) or vehicle for The diet-induced obesity in C57BL/6 mice has common characteristics 21 days. After the 15th week, all animals underwent Morris Water Maze test, of the human metabolic syndrome like high glucose concentration, insulin after which the brain was removed for studying the neuronal insulin receptor defi ciency and resistance. Studies have shown a negative relation between (IR) function, including insulin-induced long-term depression (LTD) and neu- metabolic syndrome and bone mineral density suggesting an increased ronal IR signaling, and brain mitochondrial function. In HFD rats, vildagliptin skeletal fragility. Our study aim was evaluate bone profi le in C57BL/6 male prevented peripheral insulin resistance and attenuated the impaired learn- mice induced to develop high glucose concentrations by diet-induced obe- ing and memory caused by HFD. It also restored insulin-induced LTD (Fig. A) sity, during 32 weeks. Adults C57BL/6 male mice were distributed in two and increased neuronal Akt/PKB-ser phosphorylation (Fig. B), thus indicating groups: control (n=4) and diabetic induced by high-fat diet (DH, n=4). We an improved neuronal IR function. Vildagliptin also attenuated brain mito- evaluated body weight, capillary glucose concentration, serum total ALP, chondrial dysfunction. Our fi ndings suggest that vildagliptin can reverse the tibia histomorphometric parameters [trabecular bone thickness (Tb.Th), impaired learning and memory caused by HFD, via the improved neuronal IR trabecular separation (Tb.Sp) and trabecular bone volume (BV/TV)], and function, and prevents brain mitochondrial dysfunction. mRNA expression of RANK, RANKL, OPG, PPAR-a, IL-6, TNF-_ and PTH in femur. The results showed that body weight was increased for DH (p=1.0e-3) compared to control group and similarly capillary glucose was increased in DH (p=3.0e-2) compared to control group, indicating a possible induction of insulin resistance. In bone metabolism evaluation, ALP activity for DH was increased compared to control group (p=2.4e-3) and histomorphometric pa- rameters showed a possible bone loss through decrease of Tb.Th (p=2.0e-3) and BV/TV (p=1.0e-3) on DH in relation to control group and increased Tb.Sp on DH compared with control group(1.6e-2). In addition, increased expression of RANK and RANKL associated to increased pro-infl ammatory cytokines such as IL-6 and TNF-_ in DH compared to control group (p=4.0e- 2;p=3.0e-4;p=1.0e-2;p=1.0e-2 respectively) suggests an unbalance on bone metabolism in favor of bone resorption associated to an onset of infl amma- tory process. The PTH, OPG and PPAR-a expression did not show signifi cant difference. These results suggest that high glucose concentration induced by high fat-diet may lead to an infl ammatory process and consequently to an Supported by: TRF-BRG 5480003 (S.C.), TRF-RTA 5280006 (N.C.) increased bone resorption. Supported by: FAPESP, CAPES, CNPq 2865-PO

Duodenal Endoluminal Barrier Stimulates Glucagon-Like Peptide 1 2867-PO Obesity Secretion, Improves Body Weight and Enhances Glucose Homeo- Consumption of a Western Diet With Casein-Whey Protein Causes stasis Obesity, Insulin Resistance and Changes in Gene Methylation Com- PUBLISHED ONLY OMAR AL MASSADI, KRISTY M. HEPPNER, YUANGING GUO, MAARIT LEHTI, pared to Soy Protein Diet Integrated Physiology/ JOSE BERGER, JENNA HOLLAND, NICKKI OTTAWAY, SARAH AMBURGY, CHRIS- TIMOTHY D. HOWARD, COLETTE KIRK EL-AMIN, LI ZHANG, MARIO MEDVE- TINE RAVER, DIEGO PEREZ-TILVE, PAUL PFLUGER, MATTHIAS H. TSCHOP, RANDY DOVIC, JING CHEN, SHUK-MEI HO, JANICE D. WAGNER, Winston-Salem, NC, J. SEELEY, KIRK M. HABEGGER, Cincinnati, OH, Munich, Germany Cincinnati, OH Today’s energy-dense diets and sedentary lifestyles have accelerated Nutritional interventions are important for reducing obesity and related the incidence of obesity and its comorbidities. To date, surgical interven- conditions. Soy is a good source of protein and has been shown to affect tions are the most successfully approaches to treat obesity and diabetes. plasma lipids, body weight, and insulin action. A breeding colony of cyno- However, current bariatric interventions are highly invasive and often carry molgus monkeys has been fed diets equivalent in macronutrients except signifi cant risk and thus, the development of less-invasive procedures is for the protein source. Both diets had moderate fat and cholesterol similar highly desirable. The duodenal-endoluminal sleeve (dELS) is a fl exible tube to the typical American diet and either protein derived from soy or casein- that acts as a barrier to nutrient-tissue interaction along the duodenum. Im- whey. Adults and infants born into the colony have been followed over a

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A709 OBESITY—HUMANCATEGORY

four-year period to determine the effect of diet on body weight and glyce- among diabetic patients. The prevalence of BED in ND patients (35%) was mic control. In addition, muscle biopsies were taken from offspring (n=31) greater than in diabetic patients (14.8%) (P=0.006). When we analyzed the to determine levels of gene methylation (Illumina HumanMethylation27 and patients who positively scored for BED, regardless of whether they had 450 BeadChip) and expression (Affymetrix GeneChip Rhesus Macaque Ge- DM2 or not, and compared them with the group without BED, BED patients nome Array). Adults and offspring consuming casein-whey had signifi cantly were younger (47±10 vs 54±10 years; P=0.0002), had higher body mass index increased body weight, insulin concentrations, and impaired glucose clear- (34.5±7.6 vs 31.8±5.4 kg/m2; P=0.03) and presented levels of LDL cholesterol ance. Muscle biopsies revealed 7 GO categories that were enriched for genes 27.3% greater than the patients without BED (140 ± 43 vs 110 ± 36 mg/dL; with signifi cantly increased methylation with soy, and 23 that were enriched P<0.001). No statistically signifi cant difference was observed in relation to for genes with decreased methylation with soy. Of particular interest, two the presence of BED according to the type of antihyperglycemic therapy. A KEGG pathways were identifi ed (sphingolipid and folate metabolism) as well signifi cant higher BMI and LDL cholesterol levels in patients with BED, treat- as increased methylation of HMGB1 (cell death and infl ammation) among ed for hypercholesterolemia as stated by the American Heart Association, monkeys eating the soy protein diet. Gene expression analysis identifi ed suggest that these individuals, in addition to the increased cardiovascular changes in a number of pathways between the two diets, including transla- risk related to the increased BMI and metabolic syndrome, have also an ad- tion, glycolysis-gluconeogenesis, and potential mediators of DNA methyla- ditional risk factor, LDL-hypercholesterolemia, which could be a determinant tion (folate biosynthesis, cysteine and methionine metabolism), which were for a more premature evolution to cardiovascular disease. all increased with the soy diet. In conclusions, this primate breeding colony will be useful to determine dietary effects on gene methylation and expres- 2870-PO sion as it relates to risk of obesity and insulin resistance as well as potential Improved Cardiometabolic Risk Factors in Overweight/Obese fetal programming in offspring born into this colony. Subjects Receiving Controlled-Release Phentermine/Topiramate Supported by: NIH (P40RR021380) (PHEN/TPM CR) LAWRENCE J. CHESKIN, CRAIG A. PETERSON, Baltimore, MD, Mountain View, 2868-PO CA Insulin Therapy Primarily Induces Subcutaneous Fat Deposition in Obesity increases the risk of cardiometabolic comorbidities, including Obese Diabetic Rats hypertension and type 2 diabetes. Obesity treatment goals should include SØS SKOVSØ, JESPER DAMGAARD, JENS LYKKESFELDT, CHRISTIAN FLEDELIUS, improving these parameters. PHEN/TPM CR led to signifi cant weight loss Denmark, Malov, Denmark (WL) vs placebo (PBO) through 56 weeks in the CONQUER study in obese Visceral adiposity is associated with insulin resistance and increased risk subjects with *2 weight-related comorbidities randomized to lifestyle in- of cardiovascular disease whereas recent data suggest that subcutaneous tervention plus PBO, PHEN 7.5mg/TPM CR 46mg (7.5/46), or PHEN 15mg/ adiposity might have the opposite effects. A common adverse effect seen TPM CR 92mg (15/92). This posthoc analysis assessed the proportion of sub- in individuals with type 2 diabetes on insulin therapy is weight gain and in- jects achieving the following composite goal at Week 56: WL >5%, systolic creased adiposity. Here, we studied how insulin therapy affects the relative blood pressure (SBP) <130 mm Hg, and HbA1c <6.5%. Also evaluated were accumulation of abdominal fat (visceral vs. subcutaneous) in streptozotocin mean percent WL (prespecifi ed primary end point), and total change in SBP (STZ) induced type 2 like diabetic high fat fed obese rats. Three weeks after and HbA1c at Weeks 16 and 56 in subjects achieving the composite goal. STZ treatment, the rats were divided into two intervention groups (n = 10) The composite goal was achieved by 14.5%, 45.9%, and 53.5% of the PBO, receiving either vehicle or NPH insulin twice daily for 3 weeks. Body weight, 7.5/46, and 15/92 groups, respectively, and these subjects improved WL, total body fat, visceral and subcutaneous abdominal fat were determined SBP, and HbA1c with PHEN/TPM CR vs PBO (Table). Discontinuation rates prior to STZ treatment, before starting therapy, and at the end of study. Total due to adverse events were 9%, 12%, and 19% for PBO, 7.5/46, and 15/92, body fat was quantifi ed using quantitative magnetic resonance. Visceral and respectively. The magnitude and direction of effi cacy and safety outcomes subcutaneous fat was quantifi ed using computed tomography. Diabetes re- in the subgroup of subjects >65 years of age were generally comparable to duced body weight (-11%), total fat (-30%), visceral (-19%) and subcutaneous the fi ndings in the overall population. These data confi rm that WL can drive fat mass (-47%) while insulin therapy had the opposite effect (+15%, +70%, improvements in cardiometabolic comorbidities. Rates of early goal achieve- +25%, +126%, respectively). Of the diabetes induced weight loss, 38% could ment indicate that PHEN/TPM CR has the potential to signifi cantly increase be explained by changes in adiposity whereas 51% of the weight gain fol- the number of subjects reaching broad treatment goals across many cardio- lowing insulin therapy was due to increased fat mass.This study shows that metabolic risk factors. 3 weeks of insulin therapy affects adipose tissue depots differentially. Thus, accumulation of fat occurred preferentially in the subcutaneous adipose tissue over the visceral adipose tissue. The subcutaneous adipose tissue seems to be more sensitive to changes in circulating insulin levels compared to visceral fat. Furthermore, insulin therapy increases body weight, total body fat, visceral and subcutaneous fat mass in the diabetic STZ rat. As- suming an inverse causal link between subcutaneous adiposity and insulin resistance, our results suggest that weight gain following insulin therapy in type 2 diabetics is associated with improved insulin sensitivity.

OBESITY—HUMAN

2869-PO Positive Screening for Binge Eating Disorder: Association With Changes in the Metabolic Profi le of Patients With Type 2 Diabetes Supported by: Vivus, Inc. Obesity and Overweight/Obesity HELENA SCHMID, JULIANA K. CHAGAS, MARIA C. RIOS, CLÁUDIO F. GOELZER NETO, Porto Alegre, Brazil 2871-PO PUBLISHED ONLY Integrated Physiology/ The aim of the present study was to compare patients who differed in Basal-Plus Regimen of Insulins Glargine and Glulisine Is Weight their responses to a standardized screening questionnaire for binge eating Neutral Across BMI Groups disorder (BED) for their metabolic profi le and to evaluate the possibility of an MARK R. LANKISCH, DAVID R. OWENS, EDWARD WANG, JAY LIN, STEFANO effect of different antihyperglycemic regimens upon the responses to the DEL PRATO, Düsseldorf, Germany, Cardiff, United Kingdom, Bridgewater, NJ, Flem- same questionnaire in diabetic patients. Overweight or obese patients (162 ington, NJ, Pisa, Italy type 2 diabetic (DM2) and 40 non-diabetic (ND) were recruited to participate Insulin glulisine is a novel analogue of human insulin designed for use as a in the study. The presence and severity of binge eating disorder (BED) or its rapid-acting insulin. This study evaluated the weight change resulting from absence (No-BED) was determined with the Binge Eating Scale (BES). The combining insulin glulisine with baseline insulin glargine in patients with type metabolic profi le was determined by measures of plasma glucose, glycated 2 diabetes among 3 different body mass index (BMI) ranges. This analysis hemoglobin, total cholesterol and triglycerides, anthropometric character- pooled data from 4 randomized, multicenter studies that included patients istics, the type of antidiabetic therapy and information on diabetes history with poor glycemic control who were initiated on a combination of basal

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A710 OBESITY—HUMANCATEGORY insulin glargine plus insulin glulisine for up to 6 months. A1C and demograph- fore and after 10 weeks of lifestyle modifi cation. Biochemical and anthropo- ics were measured at baseline and endpoint as specifi ed in the respective metrical measurements were performed.The life style modifi cation program trials. A total of 711 patients were included in this analysis, of whom 46.7% consisted of a reduction of energy intake to an age-adjusted optimum and an were female, and the mean age was 59.9 years. The patients were grouped exercise program (four units per week). RESULTS: The mean weight loss was by BMI < 30 kg/m2 (mean 27.0 ± 1.8), BMI 30-35 kg/m2 (mean 32.3 ± 1.5), and 3.2 ± 3.5 kg (3.7 ± 4.0%).Thr/Thr homozygotes (N=52) increased lean muscle BMI * 35 kg/m2 (mean 40.0 ± 5.4). Overall, mean A1C at baseline decreased mass (p<0.05),decreased total body fat (kg) (p<0.005) and increased basal signifi cantly from 7.6 ± 0.9% to 7.1 ± 0.9 % at endpoint (P < 0.001); 1.7% of metabolic rate (BMR) per 1 kg body weight (p<0.005) in comparison with the patients experienced a severe hypoglycemic event by endpoint. There were Met carriers (N=52), but an association between BMI decrease and AHSG no signifi cant changes in mean weight between baseline and endpoint in variants was not found. CONCLUSION: AHSG gene variants modify the ef- any of the BMI groups (BMI < 30: 75.6 vs 76.4 kg, P = 0.38; BMI 30-35: 91.3 fect of physical activity on BMR.Carriers of the Thr248Thr genotype showed vs 91.8 kg], P = 0.64; BMI * 35: 112.8 vs 114.2, P = 0.50). Overall, there was a higher benefi t from the lifestyle intervention (expressed as changes in no signifi cant difference in weight gain between the BMI groups (BMI < 30: body fat, active muscle and basal metabolic rate). 6 [SD] 0.87 kg [3.69]; BMI 30-35: 6 [SD] 0.54 kg [3.52]; BMI * 35: 6 [SD] 1.19 Supported by: No. 00023001 (IKEM, CR) kg [4.84], P = 0.27). In this analysis of 4 randomized trials, the initiation of a basal-plus regimen with glargine + glulisine was weight neutral in patients 2874-PO across a range of BMI. This basal-plus regimen signifi cantly reduced A1C, and was associated with a low prevalence of severe hypoglycemia. Supported by: sanofi -aventis WITHDRAWN 2872-PO

2875-PO WITHDRAWN Hyperinsulinemia Acutely Decreases Serum Retinol Binding Pro- tein 4 (RBP4) Concentration in Lean and Obese Subjects MAREK STRACZKOWSKI, MONIKA KARCZEWSKA-KUPCZEWSKA, AGNIESZKA NIKOLAJUK, AGNIESZKA ADAMSKA, NATALIA MATULEWICZ, MAGDALENA ZIELINSKA, MARIA GORSKA, IRINA KOWALSKA, Bialystok, Poland, Olsztyn, Poland Retinol binding protein 4 (RBP4) is a protein secreted by hepatocytes and by adipose tissue, which might induce insulin resistance by stimulating he- patic gluconeogenesis and inhibiting insulin signalling in muscle. Circulating RBP4 is increased in obesity, type 2 diabetes and coronary heart disease, however, the regulation of its level by hyperinsulinemia remains unclear. The aim of the present study was to assess the effect the effect of acute hy- perinsulinemia on serum RBP4 concentration in young healthy subjects. The study group consisted of 64 male subjects (mean age, 23.27±2.55), 34 lean (BMI below 25 kg/m2) and 30 with overweight or obesity (BMI between 25 and 40 kg/m2). Serum RBP4 concentration was measured before and after 2 hour euglycemic hyperinsulinemic clamp. Obese subjects had lower insulin sensitivity (p=0.006) and higher serum RBP4 concentration (p<0.0001). Two- hour hyperinsulinemia resulted in a signifi cant decrease in serum RBP4, it was observed in the entire study population and in the lean and obese sub- jects analyzed separately (all p<0.0001). The decrease in RBP4 in response to insulin (6RBP4) was comparable between the groups and after the clamp serum RBP4 was still higher in the obese subjects (p=0.005). Baseline RBP4 was positively related to BMI (r=0.40, p=0.001), waist circumference (r=0.42, p=0.001), triglycerides (r=0.47, p<0.001) and gamma-glutamyl-transpepti- dase activity (r=0.34, p=0.007) and negatively to insulin sensitivity (r=-0.33, p=0.007). Our data show that hyperinsulinemia acutely decreases serum RBP4 in lean and obese humans. Supported by: Grant UDA-POIG.01.03.01-00-128/08; from the Program Innovative Economy

2876-PO Breath/Plasma Gas Ratio as a Biomarker of Altered Metabolism in Obesity and Type 2 Diabetes YU-HSIN HUNG, STACY R. OLIVER, HYUN JI LEE, BRIAN D. TRAN, PIETRO R. GALASSETTI, DONALD R. BLAKE, Irvine, CA Analysis of volatile organic compounds (VOCs) in human breath, an emerg- ing novel screening/monitoring technique for endogenous metabolism, cur- Obesity 2873-PO rently implies that similar exhaled gas levels refl ect similar metabolic condi- An AHSG Gene Variant Modulates Basal Metabolic Rate and Body tions (i.e. a constant breath/plasma ratio, B/P). If this ratio were altered, PUBLISHED ONLY Composition Development after a Short-Time Lifestyle Intervention identical breath concentrations could refl ect different conditions, rendering Integrated Physiology/ Obese Females simple breath analysis inadequate to separate them. We hypothesize that in PAVEL SUCHANEK, IVANA KRALOVA LESNA, VERA LANSKA, JAROSLAV HU- obesity (OW) and diabetes (T2DM), B/P of key exhaled VOCs is altered, pro- BACEK, Prague, Czech Republic viding metabolic information not obtainable from breath analysis alone. In 7 The AHSG (alpha2 Heremans-Schmid glycoprotein) gene is suggested to healthy (CTL, 21±2yrs), 6 OW (5M, 38±6yrs) and 9 T2DM (6F, 45±3yrs) adults, be important for the regulation of body fat and insulin sensitivity. Our study we therefore studied B/P from matched blood and breath samples. Gases aimed to investigate the relationship between the common Thr248Met dissolved in plasma were extracted with a constant fl ow of helium micro- (rs4917) variant and obesity characteristics after an intervention with over- bubbles; all gases were quantifi ed by multi-column/multi-detector gas chro- weight/obese females. MATERIAL AND METHODS: We analyzed 105 unre- matography. Of ~75 VOCs analyzed, several displayed different B/Ps among lated overweight/obese nondiabetic Czech adult females (49.0 ± 12.1 years, groups; among gases related to oxidative stress and carbohydrate metabo- BMI over 28.6 kg/m2, mean BMI before intervention 32.8 ± 4.1 kg/m2) be- lism, isoprene and acetone were signifi cantly higher and lower, respectively,

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A711 OBESITY—HUMANCATEGORY

in OW and T2DM as compared to CTL; MeONO2, previously shown to track 2878-PO glucose fl uctuations in diabetes, was only elevated in OW. Our results are Laparoscopic Sleeve Gastrectomy Decreases Proinfl ammatory likely to 1) enhance the accuracy of current breath-based predictive mod- State and Reduces Chemoattracting Ability of Subcutaneous Adi- els for plasma variables related to OW and T2DM, and 2) generate novel pose Tissue in Morbidly Obese Non-Diabetic Subjects information, not obtainable via current testing methodology; together may MILOS MRAZ, JANA DRAPALOVA, ZDENKA LACINOVA, DENISA HALUZIKOVA, improve diagnostic accuracy in the early stages of dysmetabolic states, as PETRA KAVALKOVA, PAVEL TRACHTA, MARTIN MATOULEK, MOJMIR KASAL- well as monitor disease progression and effi cacy of therapeutic strategies. ICKY, STEPAN SVACINA, MARTIN HALUZIK, Prague, Czech Republic Low-grade infl ammation is one of the key mechanisms of obesity-related complications including type 2 diabetes mellitus and cardiovascular diseas- es. The objective of our study was to prospectively assess the effects of laparoscopic sleeve gastrectomy (LSG) on metabolic and systemic infl amma- tory parameters and expression of proinfl ammatory factors in subcutaneous adipose tissue (SCAT) of morbidly obese non-diabetic subjects. Eleven obese females undergoing LSG were included into the study. Anthropometric, bio- chemical and hormonal parameters were assessed before and 6, 12 and 24 months after LSG. mRNA expression of 39 adipokines and infl ammatory factors in SCAT was measured on a 7900HT Fast Real-Time PCR System in- strument using TaqMan®Custom Array. LSG markedly reduced body weight (BMI at month 24: 33.2 ± 1.9 vs. 43.2 ± 1.7 kg/m2) and improved parameters of glucose and lipid metabolism. Systemic infl ammatory markers were sig- nifi cantly decreased along with circulating leptin and insulin, while serum Supported by: NIH (1UL1RR031985; K24 DK085223) adiponectin considerably increased. Subcutaneous adipose tissue showed a substantially reduced proinfl ammatory state at 12 as well as 24 months 2877-PO after LSG with decreased expression of macrophage surface marker CD68, Differential Weight Loss Effect on Visfatin and Other Adipokines chemokines CCL-3 and -5 and chemokine receptors CCR-1, -4 and -5. mRNA and Markers of Subclinical Infl ammation expression of other proinfl ammatory cytokines, adipokines and receptors EWA KWIECINSKA, ELEKTRA SZYMANSKA-GARBACZ, ANNA BORKOWSKA, including leptin, resistin, IL-8, IL-18 and toll-like receptor 2 showed similar MALGORZATA SARYUSZ-WOLSKA, MACIEJ PAWLOWSKI, JERZY LOBA, LESZEK trends, while the expression of adiponectin and vascular endothelial growth CZUPRYNIAK, Lodz, Poland factor A markedly increased. We conclude that LSG in obese non-diabetic The effect of weight loss on recently discovered adipokines and markers subjects signifi cantly reduces body weight and improves allover metabolic of subclinical infl ammation (SI) is unclear. Fifty overweight subjects (mean and proinfl ammatory profi le. These effects might be at least partially ex- age [±SD] 53.6±12.0 years), without known diabetes were randomized into plained by decreased expression of chemotactic and infl ammation-related Group 1 undergoing intensive intervention consisting of monthly offi ce visits factors and potentially increased angiogenic capacity in subcutaneous adi- with standard lifestyle education, and Group 2 (controls) were subject to pose tissue. standard care i.e. two offi ce visits 3 months apart with identical lifestyle Supported by: MZOVFN2005 education. Oral 75 g glucose tolerance test with fasting plasma visfatin, adi- ponectin, interleukin-6, interleukin-1beta, thrombomodulin, TNF, endothelin, 2879-PO adhesion molecules, and E-selectin measurements were performed. Group 1 A Five Year Chinese Laparoscopic Adjustable Gastric Band Experi- subjects lost 9.3±5.4, and Group 2 3.6±3.1 % of baseline body weight (p<0.01 ence between groups), and proportional improvement in blood glucose and blood QIAOJUN HAN, YUE CHEN, JIANBING ZHUGE, DAJIN ZOU, ZHAOLONG ZHANG, pressure (BP) was noted. Plasma visfatin and E-selectin decreased in both Shanghai, China groups, however at a similar rate (in Group 1 from 106.2±13.7 to 47.8±21.5 Laparoscopic adjustable gastric banding (LAGB) has been our choice op- ng/ml and from 36.3±23.4 to 14.6±11.7 ng/ml; in Group 2 from 105.7±19.9 eration for morbid obesity since 2003. Despite a long list of publications to 55.7±24.1 ng/ml and from 28.3±16.9 to 12.1±9.1 ng/ml, respectively; all about the LAGB during recent years, the evidence with regard to outcome p<0.01 vs baseline). No signifi cant changes were found in other biomarkers. after LAGB in China which has a large population of obese patients has been Interestingly, there was a negative correlation between the degree of sys- rather sparse. The aim of this study was to review 5 years of laparoscopic tolic BP reduction and plasma visfatin decrease (r=0.21, p<0.05, see Figure). adjustable gastric band procedures in a single institution in Shanghai, China. In conclusion, visfatin and E-selectin seem to be the biomarkers which are All LAGB cases at the authors’ center between June 2003 and November most closely related to weight loss and subsequent metabolic improvement. 2009 were retrospectively analyzed. A telephone survey of these patients However, establishing full role of visfatin in metabolic regulation requires was also conducted in 2010. 188 consecutive Chinese patients (69.7% fe- further studies. male) underwent LAGB. The mean age was 27.2±9.1 years (range, 14-55), the mean weight was 106.8± 24.7 kg (range, 67-229.8), and the mean BMI was 37.5± 6.2 kg/m2 (range, 26.1-61.7). Only 8 patients (4.3%, 8/188) were super-obese (BMI> 50 kg/m2). The mortality rate was 0%. Six (3.2%, 6/188) bands had been removed, and 4 for band slippage (2.1%, 4/188). One case was converted to open technique. Of 188 LAGB Chinese patients, 98 were telephone surveyed. There were no signifi cant differences of preoperative characteristics between telephone surveyed and non-telephone surveyed LAGB patients. Follow-up averaged 23.6 months. Weight loss averaged 17.6 ± 12.5 kg. Percent excess weight loss (EWL) was 27.8 ± 16.4%, 39.0 ± 23.1%,

Obesity 44.1± 27.3 and 43.1 ± 28.4% at 3 months, 6 months, 1 year, and 2 years, respectively. The non-responder rate (EWL<30%) at 2 years follow-up was 33.3% (20/60). Weight regain of more than 10 kg from nadir was observed PUBLISHED ONLY

Integrated Physiology/ in 10 (10.2%) of the 98 telephone surveyed patients after LAGB. The LAGB technique is a relatively safe procedure with few major complications. Few LAGB patients underwent reoperation for complication, and mortality was zero. However, there was also a minority of morbidly obese who did not benefi t enough from this kind of bariatric surgery. Supported by: Medical University of Lodz, Poland, Grant No 502-18-835

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A712 OBESITY—HUMANCATEGORY

2880-PO Pbo N=252 Lor QD N=95 Lor BID N=256 How Can a Primary Care Physician Easily Modify Lifestyle of Over- weight Subjects Who Are At Risk of Diabetes? Change in Mean Drug n (%)1 Mean (sd) n (%)1 Mean (sd) n (%)1 Mean (sd) EWA KWIECINSKA, ELEKTRA SZYMANSKA-GARBACZ, ANNA BORKOWSKA, Dose MALGORZATA SARYUSZ-WOLSKA, MACIEJ PAWLOWSKI, JERZY LOBA, LESZEK HbA1C Change -0.4 (0.9) -1.0 (0.9) -0.9 (0.9) CZUPRYNIAK, Lodz, Poland % Daily Dose Change Prevention of type 2 diabetes is of utmost importance for every over- Metformin 229 (91) 6.6 (40.1) 88 (93) 3.0 (36.6) 236 (92) -0.8 (35.9) weight individual. Majority of the at-risk subjects are not seen by an endocri- nologist, but by a primary care physician (PCP). We conducted a three-month SFU 127 (50) 6.5 (98.9) 47 (50) -24.6 (58.0) 129 (50) -16.0 (63.0) prospective randomized controlled study assessing the effect of a simple TZD 63 (25) 3.3 (89.0) 27 (28) -21.3 (57.9) 59 (23) -16.4 (40.3) intervention possible to perform by PCP. Fifty overweight subjects, with- DPP-IV Inhibitor 37 (15) -6.9 (34.1) 12 (13) -16.7 (38.9) 23 (9.0) -4.3 (20.9) out any known glucose intolerance or diabetes attending a single PCP offi ce % of Patients: n (%) n (%) n (%) were randomized into Group 1 undergoing intensive intervention consisting of monthly offi ce visits (four visits in total) with standard lifestyle education, Stopping Drug and Group 2 (controls) - who were subject to standard care i.e. two offi ce Metformin 0 (0.0) 2 (2.1) 10 (3.9) visits three months apart with identical lifestyle education. Oral 75 g glu- SFU 8 (3.2) 13 (14.0) 21 (8.2) cose tolerance test with insulin and GLP-1 measurements were performed TZD 4 (1.6) 8 (8.4) 8 (3.1) at baseline and after the follow-up. 46 subjects (mean age [±SD] 53.6±12.0 years) completed the study. Group 1 subjects lost 9.0±5.7, and Group 2 - DPP-IV Inhibitor 3 (1.2) 2 (2.1) 1 (0.4) 3.3±3.2 kg, blood pressure decreased in Group 1 and 2 from 135±10/85±11 Starting New Drug to 125±9/73±10 and from 134±11/81±10 to 129±9/75±11 mmHg, respectively Metformin 3 (1.2) 1 (1.1) 3 (1.2) (p<0.01 vs baseline). Percentages of subjects with normal glucose tolerance SFU 10 (4.0) 3 (3.2) 9 (3.5) /abnormal glucose tolerance/diabetes changed from 10/53/37 to 42/42/16% in Group 1 and from 32/58/10 to 27/68/5% in Group 2 (p<0.05). Other results TZD 9 (3.6) 1 (1.1) 3 (1.2) are shown in Table 1. In conclusion, just doubling the number of offi ce visits DPP-IV Inhibitor 13 (5.1) 3 (3.2) 10 (3.9) containing healthy lifestyle advice led to impressive improvement in meta- 1n (%) of patients taking drug class at Baseline bolic disorders. This benefi t was largely achieved via reduction in insulin resistance rather than via enhancing incretin effect. 2882-PO Comparative Effi cacy of Lorcaserin for Weight Loss in High Risk Obese Patients and Low Risk Overweight Patients SCOTT STUBBE, JINKUN ZHANG, MATILDE SANCHEZ, WILLIAM R. SHANAHAN, CHRISTEN M. ANDERSON, San Diego, CA Lorcaserin (Lor) is an investigational agent for weight loss that acts as a selective 5-HT2C agonist. Lor was studied in 3 randomized, placebo-con- trolled trials of obese and overweight patients, whose treatments included Lor 10 mg BID or placebo (Pbo), and behavioral modifi cation counseling for 1-2 years. The purpose of this post hoc analysis was to evaluate whether Lor’s effi cacy at 1 year extended to both high risk and lower risk patients. A meta-analysis of the BLOOM, BLOSSOM and BLOOM-DM data defi ned high risk subgroups with BMI *35 kg/m2 and/or 1 or more weight related co-morbid condition (hypertension, dyslipidemia, type 2 diabetes, cardiovas- cular disease, impaired glucose tolerance), and lower risk subgroups with BMI )30 kg/m2 and 0 or 1 weight-related co-morbid condition. Lor was as- sociated with greater mean weight loss than Pbo at 1 year in each risk sub- group (Table). The greatest incremental weight loss relative to Pbo occurred Supported by: Medical University of Lodz, Poland, Grant No 502-18-835 in the lower risk groups, while the lowest incremental weight loss occurred in patients with type 2 diabetes and those with BMI*40. The meta-analysis shows that Lor causes greater weight loss than Pbo in patients with or with- out weight-related co-morbid conditions regardless of degree of obesity. 2881-PO Weight Change (%) from Placebo Lor Lorcaserin Improved Glycemic Control and Decreased Use of Anti- Baseline to Week 52 diabetic Medications during a 1-Year Study of Weight Loss in Pa- n Mean n Mean (ITT/LOCF) (sem) (sem) tients With Type 2 Diabetes EMELYN VARGAS, CINDY TURNER, MATILDE SANCHEZ, WILLIAM R. SHANA- Overall population* 1541 -2.8 (0.1) 1560 -5.9 (0.2) HAN, CHRISTEN M. ANDERSON, San Diego, CA Higher Risk Group, 248 -1.5 (0.4) 251 -4.5 (0.4) Lorcaserin (Lor) is an investigational selective 5-HT2C agonist for weight type 2 diabetes (BLOOM-DM)* management. Lor was evaluated for weight loss in 603 modifi ed intent to Higher Risk Groups; non-diabetic (BLOOM + BLOSSOM) treat patients with inadequately controlled type 2 diabetes (BLOOM-DM Hypertension* 699 -3.2 (0.2) 703 -6.5 (0.3) trial). Lor was associated with greater weight loss and improved glycemic control relative to placebo (Pbo). This analysis evaluated changes in use of Dyslipidemia* 934 -3.0 (0.2) 965 -6.8 (0.2) anti-diabetic agents during the 1 year trial. Patients received Lor 10 mg QD, Hypertension and Dyslipidemia 332 -3.4 (0.3) 349 -6.8 (0.4) Obesity Lor 10 mg BID or Pbo, and behavioral modifi cation counseling. Anti-diabetic BMI >35 to )40* 1042 -2.1 (0.2) 1083 -5.8 (0.2) drug doses were monitored. Changes from baseline to study exit were cal- PUBLISHED ONLY BMI *40* 609 -2.7 (0.3) 638 -5.3 (0.2) Integrated Physiology/ culated by drug class and total daily dose. Lor BID was associated with mean decreases in daily doses of all anti-diabetic drug classes, while daily dose Hypertension AND BMI *35 406 -3.2 (0.3) 396 -6.4 (0.3) increased in all classes except DPP-IV inhibitors with Pbo. More patients on Dyslipidemia AND BMI *35 462 -2.8 (0.3) 474 -6.5 (0.3) Lor than Pbo stopped drugs in each class except DPP-IV inhibitors. Similar % Lower Risk Groups (non-diabetic) of patients on Lor and Pbo started metformin; all other classes were initiated BMI )30 and No Co-morbidity 23 -1.1 (1.3) 12 -5.5 (2.5) less often in the Lor groups. Lor was associated with greater weight loss and improved glycemic control, with decreased use of anti-diabetic agents BMI )30 with 1 Co-morbidity 76 -2.5 (0.7) 97 -7.6 (0.7) compared to Pbo in a 1-year study. *LS mean values

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A713 OBESITY—HUMANCATEGORY

2883-PO 2886-PO Infl uence of Gastric Bypass on the Absorption of Iron Serum IL-18 Concetration is Associated With Resting Energy Ex- INA GESQUIERE, VEERLE FOULON, MATTHIAS LANNOO, BART VAN DER penditure Independently of BMI SCHUEREN, Leuven, Belgium AGNIESZKA NIKOLAJUK, AGNIESZKA ADAMSKA, MONIKA KARCZEWSKA- Roux-en-Y gastric bypass (GBP) is proposed as a treatment for type 2 KUPCZEWSKA, NATALIA MATULEWICZ, MAGDALENA ZIELINSKA, MARIA GÓR- diabetes, but may reduce the absorption of nutritional components such as SKA, IRINA KOWALSKA, MAREK STRACZKOWSKI, Bialystok, Poland iron. Aim of this study: determine the incidence of iron defi ciency (ferritin Numerous studies indicate an association between low-grade chronic ) 20 μg/L) after GBP; explore associations between post-GBP iron defi ciency infl ammation and predisposition to type 2 diabetes and atherosclerosis. IL- and gender, age, preoperative iron status and other vitamin defi ciencies; and 18 is a proinfl ammatory cytokine with proatherogenic properties. Existing examine the infl uence of GBP on the absorption of oral iron supplements. evidence indicated that circulating IL-18 was associated with insulin resis- A monocentric retrospective cohort study of patients who underwent GBP tance, metabolic syndrome and type 2 diabetes. Some data suggest that between 06/’06 and 11/’10 was conducted. In 11 patients with post-GBP iron endogenous IL-18 signaling modulates food intake, metabolism and adipos- defi ciency, an oral challenge test with 100 mg FeSO4.7H2O was performed. ity, as homeostatic regulator. The aim of the present study was to estimate The intestinal absorption of iron was measured as the difference between the relationship between seum IL-18 concentration and resting energy ex- the highest concentration of serum iron obtained 1, 2 or 3h following the penditure (REE). Our study involved 60 young (age: 23.43±2.62 years), ap- ingestion of iron, and the concentration of serum iron at t=0. Cut-off of 80 parently healthy men with normal glucose tolerance. Anthropometric mea- μg/dL was used for suffi cient iron absorption. For the analysis of the data we surements, blood biochemical analysis, euglycemic hyperinsulinemic clamp used generalized linear mixed models. Analyses were performed using NL- and indirect calorimetry before and during the clamp were performed in the MIXED procedure in SAS. The retrospective cohort consisted of 123 female studied group. The serum concentrations of IL-18 was 259.95±98.36 pg/ml. and 41 male patients with a mean age of 43±10 years. Ferritin concentra- We found that serum IL-18 positively correlated with BMI (r=0.26, p=0,04), tion pre-GBP was 106,53±77,84 μg/L. Post-GBP, 52 (42.28%) female patients fasting serum insulin (r=0.26, p=0.047) and free fatty acids after the clamp and 9 male (21.95%) patients developed iron defi ciency. Incidence of iron (r=0.50, p=0.001). A signifi cant association between basal and post-clamp defi ciency was signifi cantly lower in males than females (p=0.017). Young REE with serum IL-18 (r=0.40, p=0.001 and r=0.26, p=0.043, respectively) age (p=0.012), preoperative poor iron status (p=0.0004) and vitamin B12 defi - was observed in the studied group. In multiple regression analysis the rela- ciency (p=0,0009) were associated with a higher incidence of iron defi ciency tionship between IL-18 and REE was independent of BMI (`=0.22, p=0.008). post-GBP. Iron defi ciency seemed to increase over time (p<0.0001). In the Our data indicated that serum IL-18 concentration is associated with resting oral challenge test, there was only one patient who showed suffi cient iron energy expenditure independently of BMI. absorption. Difference in serum iron concentration was lower than 20 μg/dL Supported by: Grant: UDA-POIG.01.03.01-00-128/08 Program Innovative Economy in 2 of 11 patients. Iron defi ciency is frequent post-GBP. Female gender, 2007-2013 young-age, pre-operative poor iron status, vitamin B12 defi ciency and the period post-GBP predispose to develop iron defi ciency. Absorption of oral 2887-PO iron supplements is impaired post-GBP. Timing of Adjustable Gastric Banding and “Remission” or Improve- Supported by: Agency for Innovation by Science and Technology, Flanders ment of Type 2 Diabetes and Associated Risk Factors HELMUTH BILLY, SUNIL BHOYRUL, TED OKERSON, CHRISTOPHER CORNELL, Ven- 2884-PO tura, CA, Del Mar, CA, Yorba Linda, CA, Irvine, CA Genetic Susceptibility, Birth Weight and Obesity Risk in Young Chi- Laparoscopic adjustable gastric banding (LAGB) is one type of surgical ap- nese proach which has been demonstrated to result in signifi cant durable weight JUAN SHI, JIE HONG, LU QI, BIN CUI, WEIQIONG GU, YIFEI ZHANG, LIJUAN LI, loss; however, longer-term data in patients with Type 2 Diabetes (T2D) is MIN XU, LI WANG, YING ZHAI,WITHDRAWN LIN MIAO, RUI WANG, YUFANG BI, WEIQING lacking. Here we describe the 2-yr changes in T2D status and the associated WANG, NING GUANG, Shanghai, China, Boston, MA cardiovascular(CV) risk factors hyperlipidemia(HL) and hypertension(HTN) Thus far, approximately 30 loci infl uencing body mass index (BMI) and the after LAGB in the APEX trial (LAP-BAND AP® EXperience; NCT00501085), an risk of obesity have been identifi ed. In this study, we aimed to examine the ongoing 5-year, prospective, multi-center, observational study. “Remission” individual and joint associations of 23 BMI-associated loci identifi ed from of disease is defi ned as elimination of medication and “improvement” as a recent genome-wide association studies in Caucasians with obesity risk in reduction in medications. A total of 89/395 (22.5%) subjects who reported young Chinese. Birth weight refl ects prenatal metabolic adaption and has T2D at baseline had completed their 2 year visits, with 66 subjects having been related to later-life obesity risk. We particularly assessed whether suffi cient data to assess T2D status. Patients with T2D had a baseline BMI these genetic variants interacted with birth weight in relation to obesity of 43.9 kg/m2 and 95.6% of subjects had remission and/or improvement of risk. We recruited 540 young (14-30 yr) and obese patients (BMI * 30 kg/ T2D. Subjects with remission had a shorter mean duration of T2D (4 yrs; m2), and 500 age- and sex-matched normal-weight healthy individuals (BMI n=23) compared to improved (6.7 yrs; n=24), p=0.03. The group with no < 23kg/m2).We genotyped genetic variants in those 23 BMI-associated loci. change (n=3) had a mean T2D duration of 8.9 years. Baseline BMI was not Six loci, including SEC16B, GNPDA2, BDNF, FTO, MC4R and TMEM160, were different between the groups, with changes in BMI and %weight loss of signifi cantly associated with obesity risk, with odds ratio from 1.314 to -10.1/-22.8, -8.4/-18.5 and -8.0/-18.7 respectively (p=NS). Fifteen of the 66 1.701. These risk loci accounted for 4.84% of the genetic variance in obesity. T2D subjects also had evaluable HTN and HL data at 2 years; among these We created a genetic risk score (GRS) by summing the risk alleles of these (all of whom experienced T2D remission/improvement), 13 (86.6%) and 11 associated genetic variants. Prediction of obesity was signifi cantly improved (73.3%) had their HTN or HL resolve/improve, respectively. Further, those (P < 0.001) when the GRS was added to a model with age and gender, with with “remission” of all three CV risk factors had a mean duration of T2D improvement of discrimination for obesity by 2.7%. In addition, we found of 5.6 yr compared to 7.3 yrs for those who failed to have remission of all that the GRS interacted with birth weight in relation to obesity (Pinteraction< three conditions. Overall remission/improvement rates for HTN and HL were 0.001). The genetic effect appeared to be more pronounced in individuals 90.8% and 76.9% respectively. These results suggest that an early surgical with normal range of birth weight (25-75%) than those with either low (< weight loss intervention with LAGB in patients with T2D is more likely to

Obesity 25%) or high (> 75%) birth weight. In conclusion, we showed that the com- induce remission or control of the three major CV risk factors. Larger and bined genetic risk of variants identifi ed from European populations might longer term datasets are needed to evaluate the durability of these changes signifi cantly improve the identifi cation of high-risk group of obesity in young and the effect on CV outcomes. PUBLISHED ONLY

Integrated Physiology/ Chinese. For the fi rst time, we demonstrated birth weight might interact with Supported by: Allergan, Inc. genetic susceptibility in relation to obesity risk in later life, which deserves consideration in future efforts to prevent obesity. 2888-PO 2885-PO WITHDRAWN WITHDRAWN

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both positively associated with metabolic disorders (all p <0.0001) except for reduced HDL-C in women. Per unit of standard deviation increment, BMI was more strongly associated with hypertension than waist circumference in both genders; the corresponding adjusted odds ratios (ORs) (95%CI) were 1.83 (1.73-1.93) vs 1.15 (1.08-1.21) in men and 1.68 (1.60-1.76) vs 1.23 (1.17- 1.29) in women. Also, waist circumference seemed more strongly associ- ated, especially in women, with Diabetes Mellitus (DM), raised TG and re- duced HDL-C risk compared to BMI; the ORs (95%CI) were 1.48 (1.39-1.58) vs 1.16 (1.09-1.23), 1.56 (1.49-1.63) vs 1.17 (1.13-1.22), and 1.36 (1.31-1.42) vs 1.00 (0.96-1.04), respectively (Figure 1.). Increasing BMI and waist circumference convey different risk of metabolic disorders in the Chinese population.

2891-PO Very Morbidly Obese Patients have a High Prevalence of Glucose Homeostasis Abnormalities ROBERTO BARATTA, FEDERICA VINCIGUERRA, MARIA GRAZIA FARINA, CRISTI- NA PARRINO, PATRIZIA TITA, GIUSEPPINA PADOVA, RICCARDO VIGNERI, LUCIA FRITTITTA, Catania, Italy The obese phenotype is heterogeneous and very severe obesity has been hypothesized to result from a peculiar, less deleterious, metabolic back- ground. Aim of this study was to investigate the prevalence of abnormali- ties of glucose homeostasis in a cohort of patients with different degrees of obesity. The study included 938 patients consecutively recruited (males/ females: 270/668; age 37.3±0.4 years, mean±SEM; BMI 45.3±0.3 kg/m2). In- clusion criteria were BMI *30 kg/m2 and absence of a previous diagnosis of 2889-PO diabetes. Patients were assigned to three different classes of obesity: mild- Change in Predicted Cardiovascular Disease Risk after Bariatric moderate obese (MMO, BMI 30-39.9 kg/m2, n=306), morbid obese (MO, BMI Surgery in Severely Obese 40-49.9 kg/m2, n=374), and super-obese (SO, BMI *50 kg/m2, n=258). After MARIE-ÈVE PICHÉ, JULIE MARTIN, MARJORIE BASTIEN, SARAH-MAUDE CA- clinical evaluation, all patients underwent an oral glucose tolerance test RON-CANTIN, PAUL POIRIER, Sainte-Foy, QC, Canada with calculation of early-phase insulin secretion and insulin sensitivity (the To determine the impact of biliopancreatic diversion surgery (BPDS) on insulinogenic index and the insulin sensitivity index) and their interplay (the cardiovascular risk profi le and predicted cardiovascular risk in severely disposition index). All statistical signifi cances are given after adjustment for obese patients. We compared 1-year follow-up anthropometric and meta- age and gender. Insulin sensitivity deteriorated signifi cantly and progres- bolic profi les in severely obese patients who underwent BPDS (n=73) with sively (p for trend <0.0001) from MMO to MO, and SO patients, as measured controls (severely obese without surgery) (n=33). The 10-year predicted by both HOMA (3.9±0.1, 4.6±0.1, and 5.8±0.2, respectively), and Matsuda’s risk for coronary heart disease (CHD) was estimated using the Framingham index (3.6±0.1, 3.0±0.1, and 2.4±0.1). A worsening with more severe obesity risk-assessment tool. We assigned 10-year and lifetime predicted risks to was observed also for the insulinogenic index (1.7±0.1, 1.5±0.1, and 1.4±0.1) stratify subjects into different groups: high short-term predicted risk (* 10% and the disposition index (5.9±0.3, 4.3±0.3, and 2.6±0.2) (p for trend <0.001). 10-year risk or diagnosed diabetes), low short-term (< 10% 10-year risk)/low The prevalence of IFG and/or IGT increased signifi cantly and progressively lifetime predicted risk or low short-term/high lifetime predicted risk. During from MMO to MO, and SO patients (34.3%, 41.2%, and 50.0%, p for trend the follow-up period, body weight and body mass index decreased markedly <0.0001). A similar trend was observed also for the prevalence of unknown in the surgical group (-52.1 kg and -19.0 kg/m2 respectively, p<0.001) in com- type 2 diabetes (6.5%, 15.5%, and 20.5%, p for trend <0.0001). In conclusion parison to controls (-0.7 kg and -0.3 kg/m2, p=0.51). Weight loss in the surgi- super-obesity is associated with a deleterious insulin secretion and sensitiv- cal group was associated with a signifi cant reduction in HbA1C (6.2 vs. 5.1%), ity phenotype and a high prevalence of glucose homeostasis abnormalities. HOMA-IR index (61.5 vs. 9.3), all lipoprotein levels, as well as blood pressure values (p<0.001). The mean C-reactive protein levels decreased from 11.6 to 2892-PO 1.8 mg/L. The 10-year CHD predicted risk decreased by 43% in women and Clinical Effects of Roux-en-Y Gastric Bypass (RYGB) in Morbidly 33% in men, whereas the estimated CHD risk in the non surgical group did Obese Individuals: A Controlled Study in a Brazilian Multidisci- not change from baseline to follow-up. Before surgery, none of the women plinary Center With Eight Surgeons and only 18% of men showed low short-term/low lifetime predicted risk, HELENA SCHMID, CLÁUDIO F. GOELZER NETO, ANTÔNIO C. WESTON, Porto whereas a signifi cant proportion of subjects had high short-term predicted Alegre, Brazil risk (36% in women and 12% in men). Following surgery, 52% of women and The aim of this study was to compare the effects of RYGB (S group) with 55% of men have a calculated low short-term/low lifetime predicted risk, no surgical treatment (NS group) in a brazilian real world. A case-control and only 5% of women and 2% of men still showed high short-term pre- study was performed in 258 morbidly obese patients. A S group was paired dicted risk. These results highlight the benefi ts of BPDS surgery in reducing with a NS group by sex, age, body mass index (BMI) and follow-up time or eliminating cardiovascular risk factors and suggest a considerable impact (mean ± SD of 12 ± 2.8 and 12 ± 1.2 months, respectively). At baseline, S and on predicted CHD risk in severely obese. NS groups did not differ in BMI (P=0.061). On follow up, the S group showed reduction of BMI (38.1%; P<0.001), WC (28.6%; P<0.001), fasting plasma 2890-PO glucose (10.5%; P<0.001), serum LDL-cholesterol (21.9%; P<0.001), cases of Impact of Body Mass Index and Waist Circumference on Metabolic type 2 Diabetes (47.4%; P<0.001), snoring (95%; P<0.001), arthralgias (59%;

Disorders P=0.032), other clinical complaints (100%; P<0.001), and of the number (n) of Obesity XUHONG HOU, XIAOJING MA, YUNJUAN GU, JUNQIAN LU, WENYING YANG, medications (med) used (83.3%, P<0.001). The RYGB was also effective in re- WEIPING JIA, Shanghai, China, Beijing, China ducing in 85% (P<0.001) the n of patients with Metabolic Syndrome (MetS). PUBLISHED ONLY To explore the impact of body mass index (BMI) and waist circumference For the same parameters, in a similar follow up period, the NS group showed Integrated Physiology/ on metabolic disorders in Chinese adults. We analyzed the dataset of The only decrease in triglycerides levels (4.12%; P=0.047) and increase in n of China National Diabetes and Metabolic Disorders Study, which surveyed a med consumed (33.3%, P=0.027). However, the S group, compared with the representative sample of 46,239 adults aged 20 years and older from June NS one, showed decreased serum total proteins (6.80 vs 7.55 g/dL; P=0.047), 2007 to May 2008. Binary or multinomial logistic regression was conducted B12 vitamin (326 vs 420 pg/mL; P=0.035), increased protein defi ciency (8.5 vs to assess the association of metabolic disorders with the standardized 0%; P=0.01), and iron defi ciency anemia (6.2 vs 0%; P=0.005). In conclusion, variables of waist circumference and BMI. Raised triglycerides (TG) were in morbidly obese patients, RYGB causes mild vitamin and protein defi cien- diagnosed as TG * 1.70mmol/L. Reduced HDL cholesterol (HDL-C) was HDL- cies, but is highly effective in lowering the occurrence of obesity and MetS. C < 1.03 mmol/L in men and < 1.29 mmol/L in women. Multivariable regres- sion analysis revealed that after adjustment for age, region, smoking sta- tus, drinking status, mutually adjusted BMI and waist circumference were

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2893-PO 2896-PO Gender-Specifi c Relationship between Visceral Fat Volume and Cardiovascular Risk Factors in Patients With Type 2 Diabetes WITHDRAWN Among Wide Spectrum of BMI AMR MORSI, NUHA ELSAYED, ADHAM ABDEL MOTTALIB, OSAMA HAMDY, Boston, MA Increased visceral fat volume in patients with type 2 diabetes is known to be linked to infl ammation, insulin resistance and increased cardiovascu- 2894-PO lar risk. However, it is not clear if this relationship is identical in both men Anthropometric Indices of Obesity in Relation to Cardiometabolic and women. In a cross-sectional study, visceral fat volume was measured Disorders in Chinese Adults by bioelectrical impedance technique (ViScan) in 72 patients with type 2 LEI ZHANG, YANHU DONG, JUANJUAN LI, FENGHAI MA, YUAN JING, JAAKKO diabetes and different levels of fatness. The accuracy of this technique is TUOMILEHTO, QING QIAO, Qingdao, China, Weifang, China, Helsinki, Finland well validated against MRI. Thirty-seven women with BMI ranging between Anthropometric indicators such as body mass index (BMI), waist circum- 21.6 - 48.7 kg/m2 (mean 34.2 ± 5.6 kg/m2) and 35 men with BMI ranging ference (WC), waist to hip ratio (WHR), and waist to stature ratio (WSR) between 22.2 - 44.7 kg/m2 (mean 31.5 ± 5.2 kg/m2) were evaluated. We also are used to relate cardiometabolic risk. In order to identifi ed the optimal measured blood pressure, lipid profi le (LDL-Cholesterol, HDL-Cholesterol and anthropometric index in relation to diabetes, hypertension, and dyslipi- Triglycerides), microalbumin/creatinine ratio and high-sensitivity C-reactive demia, we enrolled 15268 Chinese adults aged 18-74 years (M/F: 7018/8250) protein (hsCRP) as indicators of increased cardiovascular risk. Other body who participated in a population-based cross-sectional survey conducted composition parameters including waist circumference, total body fat, trunk in Qingdao, China from 2001-2002. Information on demographic details as fat and fat-free mass were also measured in the two groups. Average age well as body weight, height, WC and HC, blood pressure (BP), fasting plasma of women was 59.1 ± 9.3 yrs, duration of diabetes 12 ± 7.7 yrs, and for men glucose (FPG), 2-h plasma glucose (2hPG), blood lipid profi le were measured. 64.2 ± 8.5 and 13.7 ± 9.5 yrs respectively. Total body fat and trunk fat were Receiver operating characteristic (ROC) curve was plotted and the optimal signifi cantly higher in women (90.9 ± 24.7 lbs versus 70.2 ± 28.3 lbs, p<0.01; cutoff point was identifi ed. Diagnostic accuracy was assessed by the area 46.3 ± 5.9 lbs versus 36.3 ± 7 lbs, p<0.001respectively) but visceral fat under the curve (AUC). Multiple logistic regression analysis was used to was signifi cantly higher in men (18.9 ± 5.7 versus 12.8 ± 3.8 arbitrary unit, examine the independent relationship between each anthropometric index p<0.001). Visceral fat in women signifi cantly correlated with hsCRP (r=0.55, and the risk of having cardiometabolic disorders. BMI, WSR, WHR and WC p=0.001) and triglycerides (r=0.39, p <0.05), while visceral fat in men did were positively associated with systolic BP (SBP), diastolic BP (DBP), FPG, not correlate with any of the measured cardiovascular risk factors. This re- 2hPG, triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) in lationship did not change after controlling for age, body weight, BMI and both men and women (P<0.01), but negatively related to high-density lipo- total body fat. This study suggests that the relationship between increased protein cholesterol (HDL-C) (P<0.01). WSR was found to have the largest AUC visceral fat volume and cardiovascular risk is gender specifi c; being stronger for diabetes (0.64, 95%CI: 0.62-0.65), hypertension (0.68, 95%CI: 0.67-0.70), in women even with little accumulation of visceral fat than in men. and dyslipidemia (0.58, 95%CI: 0.55-0.59) in both genders, followed by WC, Supported by: Joslin Diabetes Center BMI, and WHR. The optimal cut-off points of WSR for diabetes, hyperten- sion, and dyslipidemia were 0.52, 0.51, and 0.53, respectively. The odds ratio 2897-PO (OR) of WSR were 1.87 (95%CI: 1.57-2.24) in men and 1.74 (95%CI: 1.54-1.82) Effect of the Administration of Fucoidan on Anthropometric Mea- in women for having diabetes, 1.80 (95%CI: 1.52-2.11) and 2.43 (95%CI: 2.09- surements, Lipid Profi le, Insulin Resistance and Insulin Secretion in 2.82) for hypertension, and 1.40 (95%CI:1.19-1.65) and 1.31 (95%CI: 1.07-1.61) Adults With Overweight or Obesity for dyslipidemia, respectively. WSR might be the optimal anthropometric MANUEL GONZÁLEZ-ORTIZ, DIANA M. HERNÁNDEZ-CORONA, ESPERANZA index in assessing cardiometabolic risk in Chinese adults. MARTÍNEZ-ABUNDIS, Guadalajara, Mexico Overweight and obesity are associated with insulin resistance; they are 2895-PO important risk factors to develop type 2 diabetes and cardiovascular dis- Effect of Gastric Bypass on Type 2 Diabetes Patients in Obesity vs. eases. Nutritional modifi cation is the accepted indication to treat these pa- Non-Obesity in China During 3 Year Follow-Up tients, however, several adjuvant have been used for such purpose, one of HONGKAI GAO, Beijing, China them is fucoidan, a natural extract of brown seaweeds with some benefi ts Rapid improvements in glucose metabolism following Roux-en-Y gastric on health, including based on both, “in vitro”, and animal model studies, an bypass (GBP) surgery have been reported on type 2 diabetes mellitus (T2DM) improvement in weight, lipids, and insulin sensitivity. The aim of this study with obesity patients. However, data concerning non-obese Asian patients was to evaluate the effect of fucoidan on anthropometric measurements, have not been reported, and the mechanism of improving T2DM after sur- lipid profi le, insulin resistance and insulin secretion in adults with overweight gery in obesity and non-obesity population remains speculative. From Janu- or obesity. A randomized, double blind, placebo control, clinical trial was ary 2006 to June 2007, 102 T2DM patients undergoing GBP were recruited performed in 25 adults with overweight or obesity (BMI: 25.0 to 34.9 kg/m²), for our study and were divided into obesity group and non-obesity group by if without diabetes mellitus, or any pharmacological treatment. 13 patients body mass index(BMI) >25 or not. Oral glucose tolerance test, insulin release received, orally during 3 months 500 mg of fucoidan (GreenFoods) daily on test, C peptide release test, HbA1C, BMI, 6I30/6G30 and 6I30 were evalu- morning and 12 placebo. At the beginning and the end of the study anthropo- ated before and after surgery during the 3 years follow-up. After three years, metric measurements, lipid profi le, insulin resistance (HOMA IR) and insulin fasting plasma glucose returned to normal in 78% of non-obesity patients secretion (HOMA `-cell) were evaluated. Statistical analyses were calcu- and 90% of obesity patients. The HbA1C level returned to under 7.0 in 85% lated with Wilcoxon and Mann Whitney U tests. Both groups were similar of non-obesity and in 95% of obesity patients. BMI dropped signifi cantly in in basal characteristics. There were signifi cant decreasing on diastolic blood obesity group and slightly in non-obesity group. Fasting C peptide and 6I30/ pressure (72 ± 12 vs. 68 ± 14 mmHg, p = 0.041), and LDL-C levels (3.1 ± 0.5 vs. 6G30 increased signifi cantly in non-obesity group comparing with the level 2.7 ± 0.6 mmol/l, p = 0.008) after fucoidan administration, with increasing on

Obesity before surgery. About the insulin resistant, HOMA-IR decreased obviously insulin levels (60.6 ± 24.0 vs. 78.6 ± 32.4 pmol/l, p = 0.026), insulin resistance in obesity group. During the long-term follow-up, we found GBP has differ- (1.9 ± 1.2 vs. 2.8 ± 1.8, p = 0.026) and insulin secretion (35.0 ± 20.8 vs. 50.6 ± ent mechanisms on diabetes in obesity and non-obesity patients. In obesity 18.7, p = 0.033). There were no changes with placebo administration. In PUBLISHED ONLY Integrated Physiology/ group, the remission of T2DM is more related with insulin resistance while in conclusion, fucoidan during 3 months in adults with overweight or obesity non-obesity group, it associated with the improvement of islets function. decreased diastolic blood pressure and LDL-C levels, with increasing insulin Supported by: America-Asia Diabetes Research Foundation levels, insulin resistance and insulin secretion.

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A716 OBESITY—HUMANCATEGORY

2898-PO ized the effect of baseline BMI, menopausal status and age on weight loss Serum Omentin is Neither Associated With the Metabolic Syndrome in subjects who completed at least 26 days of treatment. Data are presented Nor With Angiographically Determined Coronary Artery Disease for completers treated with 0.9 mg/m2, 3 or 6 mg beloranib (beloranib N=17) ALEXANDER VONBANK, CHRISTOPH H. SAELY, AXEL MUENDLEIN, KATHRIN or placebo (N=11). A total of 34 subjects were enrolled into these treatment GEIGER, PHILIPP REIN, HEINZ DREXEL, Feldkirch, Austria, Triesen, Liechtenstein, arms. Baseline characteristics of completers (mean±SD): 93% Caucasian, Philadelphia, PA 49.2±8.9 years, 102.6±12.5 kg bodyweight, and 39.1±3.7 BMI. Weight loss Some recent small studies have decribed associations of the new adipo- was greater with beloranib (mean±SE: -4.3±0.4 kg; p<0.001, paired t test for cytokine omentin with the metabolic syndrome (MetS) and with cardiovas- change from baseline) than with placebo (0.6±0.5 kg, NS). Among beloranib- cular disease. However, data still are very scarce. We therefore measured treated subjects, starting BMI generally did not affect weight loss, although serum omentin in 395 patients undergoing coronary angiography for the subjects in the lowest baseline BMI quartile (BMI 33-38, n=5) lost slightly evaluation of established or suspected stable CAD; the MetS was defi ned more weight (mean±SE: 5.3±0.7 kg) compared to subjects in the 2nd, 3rd and 4th according to NCEP-ATPIII criteria; signifi cant CAD was diagnosed when coro- quartiles (3.8±0.7, 3.7±0.8 and 3.6±1.1 kg). Similarly, weight loss was slightly nary stenoses *50% were present. Omentin was positively correlated with greater among subjects below (4.9±0.6 kg) compared to above (3.7±0.6 kg) age (r=0.170; p <0.001) but did not show signifi cant correlations with waist the median BMI of 40. Weight loss was also similar among premenopausal circumference, fasting glucose, HDL cholesterol, triglycerides, systolic blood (4.0±0.6 kg, n=11) and postmenopausal women (4.6±0.6 kg, n=6), although pressure, or diastolic blood pressure; it was similar in MetS patients (n=118) there was a nonsignifi cant trend for women over 50 years of age to lose as in subjects without the MetS (15±21 vs. 14±15 ng/ml; p=0.460). Omen- slightly more weight (4.6±0.5 kg, n=11) than those under 50 years (3.3±0.6 kg, tin also did not differ signifi cantly between patients with signifi cant CAD n=6). The most common AEs were nausea, infusion site injury and headache. (n=190) and those without signifi cant CAD (14±19 vs. 15±15 ng/ml; p=0.233). Most were of mild/moderate intensity and tended to be self-limiting. Weight When both, MetS and CAD status were considered, omentin was similar loss with intravenous beloranib was consistent across starting BMI groups, in MetS patients as in subjects without the MetS both among those who and was generally independent of age and menopausal status. had signifi cant CAD (15±13 vs. 15±13 ng/ml; p=0.482) and among those who did not have signifi cant CAD (16±30 vs. 14±15 ng/ml; p=0.876); it further did 2901-PO not differ signifi cantly between patients with signifi cant CAD and subjects The Correlation of Eating Out With Obesity without signifi cant CAD among MetS patients (p=0.321) nor among subjects MAN-MEI SEA, JEAN WOO, Hong Kong, China without the MetS (p=0.452). We conclude that omentin is neither associated These days it is well known that obesity is a problem in developed coun- with the MetS nor with angiographically determined CAD. Omentin there- tries which is including Hong Kong. In Hong Kong, the rate of obesity stays fore does not appear to be a useful marker of cardiometabolic disease. high in the past decade. One of the causes of this might be the trend of Supported by: Research Foundation of the Austrian National Bank eating out. The aim of the study was to investigate the correlation between the frequency of eating out and obesity. The demographic parameters, body 2899-PO weight, body fat, waist and hip circumferences, blood pressure, eating out Hypolipemic Effect of Inulin Type Agave in Dyslipidemic Obese frequency and pattern were examined in a sample of 1059 subjects (mean Subjects age 39.5) enrolled in a randomized behavioral intervention trial. The results SANDRA O. HERNANDEZ, ROSA I. BRICIO, ERNESTO G. CARDONA, SARA PAS- showed that the total eating out frequency was positively associated with COE, MARIA G. RAMOS, LEONEL GARCIA, CRISTINA ISLAS, SYLVIA E. TOTSUKA, body weight (r =0.323), body fat (r=0.177), waist (r=0.185) and hip circum- TERESA A. GARCIA, Guadalajara, Mexico ferences (r=0.215). In male, the frequencies of eating out at breakfast and Dyslipidemia is a pathological condition with altered metabolism and dinner were signifi cantly positively associated with body weight, body fat, the serum lipids. Preliminary evidence in animals indicates that adminis- waist circumference and blood pressure, but the effect of eating out at lunch tration of inulin extracted from chicory root by their content of Phospho- was only positively associated with body weight. In female, the frequency oligosaccharides (POS) favorably modifi es serum lipid levels, however, re- of eating out at breakfast was signifi cantly positively associated with body sults from human studies are controversial. Agave tequilana Weber variety weight, body fat, waist and hip circumferences and systolic blood pressure, azul is an economically important in mexican people because it is the sole but the effect of eating out at lunch and dinner was only positively associat- plant allowed for tequila production but because it is a potential source of ed with body weight. Although it is inevitable to prevent people from eating prebiotics, The Inulin-type agave are nondigestible/fermentable carbohy- out, the effect of eating out at different meal time may result differently. The drates which are able through the modifi cation of the gut microbiota, the effect of taking breakfast and dinner at out is more adverse in body fatness POS content of agave inulin differs from inulin extracted from chicory root. and blood pressure in male while eating out at breakfast will be more easily with potential benefi t in cardiovascular risk factors. The aim was to assess prone to be overweight in female. the effi cacy and safety of inulin type agave on lipid profi le in dyslipidemic obese subjects. A clinical trial, open was carried out in 30 obese, hyper- 2902-PO trygliceridemic and hypercholesterolemic subjects between 18 and 60 years Prevalences of Overweight and Obesity in Chinese Adults old. All the subjects received 15 g/day of inulin in the morning, during 60 WEIPING JIA, ZHAOJUN YANG, XUHONG HOU, XIAOJING MA, SHENQI WANG, days. Biochemical and metabolic profi les before and after pharmacological WENYING YANG, Shanghai, China, Beijing, China intervention were performed. After inulin administration, there was a signifi - In the context of rapid economic and social development, there has been cant reduction of the trygliceride concentrations (197.5±72.9 and 179.1±37.0 limited report on the prevalence of obesity in China based upon national mg/dL; p=0.042). Glucose serum (83.0±13.4 and 77.6±12.9 mg/dL; p=0.007), samples in the last 10 years. This paper aimed to provide current data on the and HbA1c (5.8±0.4 and 5.7±0.3 % p=0.007). We observed a tendency to prevalence of overweight and obesity in Chinese adults. The China National increase the high density lipoprotein. There was not a signifi cant reduction Diabetes and Metabolic Disorders Study, a cross-sectional survey, was of total cholesterol, low density lipoprotein and very low density lipoprotein. conducted among a nationally representative sample of 46,239 adults aged Anthropometric parameters did not change in the group and soluble fi ber 20 years or above in 2007-2008. BMI and waist circumference were mea- intake did not produce any gastrointestinal adverse effect. The increase of sured. Overweight and obesity were defi ned according to the World Health

fi ber intake (inulin type agave) are effi cacy and safety to reduced trygliceride Organization (WHO) criteria. According to the International Diabetes Federa- Obesity concentrations levels in dyslipidemic obese patients. tion (IDF) guidelines and Chinese Joint Committee for Developing Chinese guidelines (JCDCG), central obesity was defi ned as waist circumference * PUBLISHED ONLY 2900-PO 90 cm in men (both guidelines) and either * 80 cm (IDF guideline) or * 85 cm Integrated Physiology/ Signifi cant Rapid Weight Loss With Beloranib is Independent of (JCDCG guideline) in women. All means and prevalences were weighted and Baseline Bodyweight, Age or Menopausal Status in Women Com- standardized according to the Chinese population data in 2006. Mean BMI pleting Two Phase I Trials and waist circumference were 24.0 kg/m2 and 83.5cm for men, and 23.4 kg/ DENNIS D. KIM, JAMES E. VATH, JOE PROIETTO, JOANNE MARJASON, THOM- m2 and 78.0cm for women, respectively. The prevalences of overweight and AS E. HUGHES, Cambridge, MA, Heidelberg, Australia, Herston, Australia obesity were 30.8% and 6.0% in men, while they were 24.9% and 4.9% in Beloranib is a fumagillin class methionine aminopeptidase 2 inhibitor that women. The prevalences of central obesity based on IDF and JCDCG defi - reduces food intake and causes long term weight loss in obese animals by nition were 29.0% in men, and 41.6% and 25.2% in women, respectively. increasing triglyceride mobilization and ketogenesis. Two Phase I studies The overall prevalences of overweight, obesity and central obesity were all have evaluated the effects of twice weekly intravenous beloranib on weight higher in men than in women (all P<0.05) except for the prevalence of central loss and associated cardiometabolic benefi t in obese women. We character- obesity as defi ned by the IDF, and higher in urban areas than in rural areas

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A717 OBESITY—HUMANCATEGORY

in men (all P<0.05) but not in women. The age-specifi c prevalences of over- 2905-PO weight for both genders and central obesity for men markedly increased up The Low Quantitative Ultrasound Value at Radius is an Additional to 50-59 years of age, while the age-specifi c prevalences of central obesity Indicator for Metabolic Syndrome and Cardiovascular Disease Risk for women signifi cantly increased across all age groups, regardless of the BEI TAO, JIAN-MIN LIU, ZHENG LI, HONG-YAN ZHAO, LI-HAO SUN, LIN ZHAO, diagnosis criteria (all P<0.001 for trend test). Our results indicates that over- WEI-QING WANG, XIAO-YING LI, GUANG NING, Shanghai, China weight and obesity have become serious threats to public health in China. The objective of the present study was to determine the relationships between quantitative ultrasound (QUS) values, metabolic syndrome (MetS) 2903-PO and cardiovascular disease (CVD) risk. This was a cross-sectional study Socio-demographic, Behavioral, and Biological Predictors of conducted in Shanghai, China. One-thousand four hundred and thirty-nine Weight Loss in Pacifi c Islanders Chinese women and men with or without MetS were studied. Speed of JOSEPH K. KAHOLOKULA, CLAIRE M. TOWNSEND, ARLENE IGE, KAIMI SIN- sound (SOS) at radius, phalanx, and tibia were measured by QUS and their CLAIR, MARJORIE L. MAU, DONNA-MARIE PALAKIKO, SHERYL R. YOSHIMURA, relationships with MetS and Framingham’s 10-year cardiovascular disease ANNE LEAKE, PUNI KEKAUOHA, Honolulu, HI risk scores were evaluated. Our results demonstrated that subjects with Native Hawaiians and other Pacifi c Islanders (NHs/PIs) have a high obesity MetS had signifi cant lower SOS at phalanx, radius and tibia than those with- (49%) and diabetes (20%) prevalence compared to other ethnic groups. They out MetS (P<0.01). Adjusting for age, gender, BMI and menopausal status, face socioeconomic stressors and live in obesiogenic environments that the difference remained at radius SOS (P = 0.016). The prevalence of MetS impede their weight loss efforts. Elucidating factors related to intentional showed its increasing trend with the declining tertiles of SOS at radius. The weight loss in NHs/PIs can lead to effective interventions. We examine the middle and the lowest tertile group had odds ratio of 1.30 and 2.19 for MetS sociodemographic, behavioral, and biological factors related to *3% weight after adjustment for confounding factors (P=0.025). The radius SOS dem- loss in 100 overweight/obese NHs/PIs who completed a lifestyle program. onstrated a negative associations with BMI (r = -0.062, P = 0.023), waist A total of 56 NHs, 22 Chuukese, and 22 other PIs participated in the random- circumference (r = -0.119, P<0.001), waist-hip ratio (r = -0.149, P<0.001), 2-h ized controlled trial. All completed a 3-month weight loss program (WLP) to postprandial glucose level (r = -0.146, P = 0.003). The CVD risk increased initiate weight loss and then randomized into a 6-month family/community with decreasing tertiles of radius SOS after adjustment (P <0.001). It is thus focused weight loss maintenance program called the PILI Lifestyle Program concluded that individuals with lower radius SOS may have greater risks of (PLP; n=49), or a standard behavior weight loss maintenance program (SBP; MetS and CVD, which suggests the radius QUS value as an additional indica- n=51). We collected baseline and 3- and 9-month follow-up data on socio- tor for MetS and CVD risk. demographic characteristics, weight (kg), physical functioning (by a 6-min. Supported by: National Natural Science Foundation of China (30725037, 81000359, walk test), dietary fat, exercise frequency, weight (kg), and blood pressure. 81070693) Based on ANCOVA or logistic fi t, ethnicity, sex, initial weight loss, fat in diet at baseline, change in systolic BP, and intervention type were signifi - 2906-PO cantly associated (p).05) with *3% weight loss at 9-month follow-up. When Families United/Familias Unidas: Development and Feasibility of a entered into a logistic regression model, we found that Chuukese partici- Family-Based Group Offi ce Visit Model for the Primary Prevention pants (OR=6.04; CI=1.14-32.17) and participants who lost more weight in the of Type 2 Diabetes fi rst 3-months (OR=1.47; CI=1.22-1.86) and who were in the PLP (OR=4.50; RANDA M. KUTOB, VIOLET PEREZ-SIWIK, MARY LAREZ, MIKEL AICKIN, CHERYL CI=1.50-15.14) were more likely to achieve *3% weight loss [model: r2 (7, RITENBAUGH, Tucson, AZ N=100) = 45.50, p < .0001]. It appears that the same behavioral lifestyle Pre-diabetes affects 79 million people in the U.S. Intensive lifestyle modi- intervention does not uniformly benefi t all NHs/PIs, possibly due to differ- fi cation can delay the onset of type 2 diabetes. Families United/Families ences in acculturation status (e.g., Native vs. immigrant) and social support. Unidas was a pilot study, combining diabetes prevention with the benefi ts of The fi ndings also point to the importance of initial weight loss to sustain family support utilizing a physician-led, group offi ce visit model. The objec- motivation toward long-term weight loss maintenance. tives included adapting the Diabetes Prevention Program (DPP) for family Supported by: NIMHD of NIH (R24MD001660) physicians, collecting feasibility data, and examining the intervention’s ef- fect on diabetes risk factors in a multi-ethnic population. Adults, ages 18- 2904-PO 70, with any diabetes risk factor were eligible. Participants identifi ed one Human Adipose Tissue Macrophages Express Markers of both support person, age 14-70, to join them in the intervention. Recruitment, Classical and Alternative Activation in Obesity retention, and diabetes risk-related outcome data were measured. Program NEDA RASOULI, ROCIO PEREIRA, WILLIAM JANSSEN, Denver, CO, Aurora, CO development and feasibility data are reported here. Twelve, biweekly inter- Obesity is associated with chronic infl ammation and increased infi ltration vention sessions were developed from DPP and previously developed group of macrophages in adipose tissue, however, the function of adipose tissue offi ce visit materials. Sessions emphasized nutrition, physical activity, and macrophages (ATMs) is not fully understood. In animal models, obesity is as- resilient thinking. Primary participants plus supporters (n=58) were recruited sociated with a shift in the programming of ATMs to a classically activated from September 2009 to August 2010. The average age was 44.8±12.4; 74% state, with an increase in CD11c+ macrophages. Other surface markers such were female. Fifty-seven percent self-identifi ed as White, 37% as Latino, as CD86 and CD206 have been used as markers for classical (M1) and alterna- and 5% as African American. Being overweight/obese (72.4%) and family tive (M2) activation respectively. We hypothesized that surface markers such history of diabetes (55.2%) were the most commonly reported diabetes risk as CD206, CD11c and CD86 can differentiate human ATM activation states in factors. Fourteen (24%) were recruited, but never attended group offi ce ses- obesity. The abdominal subcutaneous adipose tissue biopsy specimens (n=4) sions. Reasons for non-participation included both family and work-related of obese non diabetic subjects were digested to obtain the stromal vascular issues. These individuals were signifi cantly younger (37.0±7.9, p=.001), more fraction. The common leukocyte antigen CD45 and the monocyte differen- often on Medicaid (p=.002), and had lower educational levels (p=.046) as tiation antigen CD14 were used to identify ATMs. Flow cytometry was used compared to participants. Of those attending at least one session, the at- to assess the percentages of CD206, CD11c, CD86 and CD11b positive cells. tendance rate was 72%. Family physicians delivering group offi ce visits to All ATMs expressed high levels of CD11b suggesting that CD11b can also be participants and their support persons proves feasible, acceptable, and has

Obesity used as a marker for macrophages. However, there were subpopulations of potential to be a sustainable model for diabetes prevention. ATMs that displayed differential expression of markers. The majority of ATMs (77%) in obesity expressed both CD206 and CD86; among this population, 2907-PO PUBLISHED ONLY Integrated Physiology/ most (70%) were CD11c negative but a small subgroup (7%) was also positive Racial Differences in the Metabolic Consequences of Abdominal for CD11c. Only a small population of ATMs (9%) was negative for CD206 (ABD) Fat Distribution in Obese Postmenopausal Women and CD86 but positive for CD11c. Furthermore, the expression of CD11b and SEAN VASAITIS, RONALD L. PRIGEON, JACOB B. BLUMENTHAL, ALICE S. RYAN, CD11c were similarly elevated in both ATMs and peripheral blood monocytes; ANDREW P. GOLDBERG, Baltimore, MD however, ATMs had signifi cantly higher levels of the expression for CD206 Excess visceral adipose tissue (VAT) is associated with metabolic dys- and CD86 when compared to monocytes. Altogether these data suggest that function, leading to the metabolic syndrome (MetSyn) and glucose intoler- ATMs are heterogeneous in regard to cell surface markers; the majority of ance (IGT). ABD subcutaneous adipose tissue (SAT) is considered metaboli- ATMs in obesity express markers for both M1 and M2. Whether CD206 and cally protective, and is higher in Black (B) than Caucasian (CAU) obese older CD86 are markers for ATM phenotype or simply markers of macrophage dif- women. We hypothesized that racial differences in the ABD distribution ferentiation from monocytes needs further investigation. of VAT and SAT affect the prevalence of MetSyn and IGT in older B and Supported by: VA Merit CAU women. DXA and CT scans (L4-5), lipid/glucose metabolic profi les and

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A718 ISLET BIOLOGY—APOPTOSISCATEGORY blood pressure (BP) were measured in 83 obese (%Fat >35%) B (48±5 %Fat, plasmic reticulum stress pathway may be a potential tool to improve islet mean±SE) and 199 CAU (47±5%, p<0.05) women 60±6 yrs. old to determine vascularization and, indirectly, islet function. the prevalence of MetSyn (ATP III criteria) and IGT (2hr glucose, G120 >140, Supported by: Ministry of Science and Technology of the People’s Republic of <200 mg/dl). A logistic regression model, controlling for %Fat, was used to China predict racial differences in the prevalence of MetSyn and IGT by VAT and the VAT/VAT+SAT(TAF) ratio. The B had greater SAT (p<0.0001), but similar 2910-PO VAT that resulted in lower VAT/TAF than CAU (0.22±0.07 vs. 0.26 vs. 0.07, Effect of Streptozotocin on Zn+2-ions in Pancreatic B-cells p<0.001). Both VAT and VAT/TAF were independent predictors of MetSyn GABIT G. MEYRAMOV, AUSULU A. KIKIMBAEVA, ASHIM B. AUBAKIROV, AISAN -10 -8 -6 -6 (p = 10 , 10 , respectively) and IGT (p =10 , 10 ). The racial difference in G. MEYRAMOVA, ASEL R. ALINA, ULMEKEN A. AKHMETJANOVA, Karaganda, predicting MetSyn was greater for the VAT/TAF ratio than VAT alone (p = Kazakhstan, Astana, Kazakhstan 0.02 vs. p=0.06). Likewise, the racial difference in predicting IGT was greater It is known that in vivo administation of Streptozotocin (SZ) to animals for VAT/TAF than VAT alone (p<0.01 vs. p=0.03). Blacks had a lower VAT/TAF results in direct selective death of B-cells. This study explores Zn+2 content than CAU for the same prevalence of MetSyn and IGT (p<0.02), i.e., a >70% in B-cells after interaction with SZ. Groups of animals: A) control: 3 rats prevalence of MetSyn and IGT occurred at a VAT/TAF of 0.27 and 0.33 for B, injected with SZ, 35-41 mg/kg - sacrifi ced after 4 days. B) 3 rats injected and 0.33 and 0.42 for CAU, respectively. There were also racial differences with SZ, 36-40 mg/kg - sacrifi ced after 15 minutes. C) 2 rabbits: injected in the relationship of VAT/TAF to the TG (p<0.01) and BP (p<0.04) constitu- with SZ, 0.4-0.5 mcg/ml, and pancreas sections assayed after 30 min. D) ents of the MetSyn, as well as G120 (p<0.04) and HOMA-IR (p<0.0001). This 6 rats injected with Glibenclamide, 1.8-2.0 mg/kg, (complete elimination of suggests the relative amount of ABD SAT alters the adverse metabolic ef- Zn+2 from B-cells) 4 days prior to administration of SZ, 36-39 mg/kg - sacri- fects of VAT in obese Black and CAU postmenopausal women. Thus, there fi ced 4 days after SZ injection. All animals maintained on non-diabetogenic may be racial differences in ABD VAT and SAT metabolism that affect the diet. Methods: Blood glucose level (BG) daily for 1 week. Insulin (IN) in B- risk for MetSyn and IGT in these obese women. cells by immunohistochemical (IG) staining; Zn+2-ion concentration with 8-para(toluenesulphonylamino)quinoline (PSQ) and vital staining with Di- thizone (DZ). Histology: Aldehyde-fucshin (AF) method. Results: A) BG rose ISLET BIOLOGY—APOPTOSIS from 4.0±0.2 mM to 12.6 ±0.7 mM; AF: destruction and death of B-cells on 70-75% of islet’s surface in 75-80% of islets; thus SZ induced diabetes. B) 2908-PO PSQ: reduced from 1.89±0.07 to 1.18±0.09; IG: 1.87±0.09 (no signifi cant dif- Pancreatic Stellate Shaped Cells are Present in Islet of a Spontane- ference from grp A); thus SZ immediately binds Zn in B-cells. C) PSQ: reduced ous Model of Type 2 Diabetes Mellitus, The GK Rat: Implications for from 2.02±0.08 to 1.09±0.04; DZ: from 1.84±0.07 to 1.11±0.04; thus SZ binds Islets Fibrogenesis in Type 2 Diabetes Mellitus Zn within B-cells rather than being eliminated by cell death. D) 4 rats: BG FENGFEI LI, ZILIN SUN, JUNMEI YIN, QING ZHAI, BIJUN CHEN, Nanjing, China slight increase from 4.2± 0.04 mM to 5.8±0.07 mM; AF: partial necrobiosis The cellular mechanisms leading to islet fi brosis in Type 2 Diabetes of B-cells on 15-20% of islet’s surface in 5-12% of islets; PSQ and DZ: from Mellitus(T2DM) remain unclear. Recent studies suggest that pancreatic stel- 1.85±0.08 to 1.49±0.18; IG: slight reduction from 1.87±0.08 to 1.67±0.09. 2 late cells(PSCs) are involved in the progression of pancreas islets fi brosis. rats: larger increase in BG to 8.8±0.11 mM after 4 days; AF: necrobiosis and The aim of ourstudy was to examine whether PSCs are present in fi brotic death of B-cells on 25-40% of islet’s surface in 35-40% of islets; PSQ and islets in spontaneousmodel of T2DM, the Goto-Kakizaki (GK) Rat. Isolated DZ: fell from 1.83±0.09 to 1.14±0.09; IG: greater reduction from 1.85±0.09 to islets from 4-month-old-male GK rat and sex-age-matched Wistar control 1.05±0.06. We conclude that diabetogenic activity of SZ may be determined rat were immunostained for a-SMA, islet fi brosis were examined by Mas- by its Zn+2 chelating ability in B-cells and that prior elimination of Zn+2 from sonstaining assay. Cells grown out from islets were determined by phase B-cells mediates this diabetogenesis. contrast,stained by immunofl uorescence microscopy for a-SMA/GFAP, a- SMA/Vimentin,a-SMA/CD31, Nestin, and be quantifi ed by real-time PCR and 2911-PO Western Blot. Masson staining showed strongly positive within fi brotic GK Effects of Xanturenic Acid on Zn+2 Content of Pancreatic B-cells islets, but not in Wistar islets. Immunofl uorescence microscopy showed that AISAN G. MEYRAMOVA, FARIDA A. MINDUBAEVA, AUSULU A. KIKIMBAEVA, a-SMA positively stained cells distribution in islet fi brotic areas. At cultured GABIT G. MEYRAMOV, Karaganda, Kazakhstan, Astana, Kazakhstan 2-3days, pancreatic stellate shaped cells (PSSCs) began growing out from is- Xanturenic Acid (XA), a diabetogenic metabolite of Tryptophan, is formed lets, more and more PSSCs grown out from GK islets, but not from the Wistar in elderly humans with vit.B6 defi ciency. In B-cells, XA forms an XA/Zn+2 islets, at the following cultured period. Cells grown out from GK islets had a complex resulting in a 49% reduction of insulin (IN) activity. This study inves- low apoptosis rate, a high proliferation rate, and could be cultured at least tigated the Zn+2 content in intact B-cells of animals with diabetes caused for 45 passages in vitro. The 3-45 passages stained positively for a-SMA, by XA. Groups of animals: A) 8 rats maintained 3.5 months on a diabetogenic GFAP, Vimentin, Desmin, Nestin, but negatively for CD31.1. PSSCs are Pres- diet, stimulating endogenous synthesis of XA (starch-52%, casein-22, but- ent in fi brotic islets of GK rat, these cells mainly distributedin fi brotic islet ter-14, yeast-3, sugar-6, salts-3). B) 6 rats treated with Glibenclamide (GB), areas. 2. These cells resembling activated PSCs, but had ahigher prolifera- 1.8 mg/ kg 3 days, resulting in complete elimination of IN/Zn+2 complex tion rate compared with PSCs isolated from Wistar rat pancreas, and could from B-cells. C) 8 rats treated with GB, 1.8 mg/kg 3 days + ordinary, non- be cultured at least for 50 passages in vitro. diabetogenic diet 14 days. Methods: Blood glucose level (BG) weekly, XA Supported by: Chinese Natural Science Foundation in urine (XAU) in Group A monthly. Pseudoisocyanine (PS) and Immunohis- tochemical (IG) staining of IN in B-cells and fl uorescent staining of Zn+2-ions 2909-PO to measure density of staining. Histology: Aldehyde-fucshin (AF) method. Hyperglycemia Induced Islet Endothelial Cell Line MS-1 Apoptosis Results. Group A: BG rose from 4.6±0.5 mM to 11.8±1.2 mM; XAU rose from Through Endoplasmic Reticulum Stress Pathway 0.044±0.08 mg/ml to 0.386±0.061 mg/ml; histology: necrosis and hydropic XUAN LIU, KUAN-FENG XU, HENG CHEN, XIN-YU XU, TAO YANG, Nanjing, China degeneration of B-cells on 60-65% of islet’s surface in 75-80% islets. In Pancreatic islet microendothelium and `-cells exhibit an interdependent destroyed B-cells: marked decrease in IN and Zn+2 content (44% and 48% physical and functional relation-ship. In this study, we investigated the ef- respectively), PS and IG dropped from 1.97±0.07 to 1.10±0.06, Zn+2 fell from fect of hyperglycemia on the apoptosis and proliferation of islet endothelial 2.02+0.07 to 1.05± 0.06. While in intact islets IG and PS fell modestly from cell line (MS-1). MS-1 cells were treated with different concentrations of 1.98±0.09 to 1.68±0.11 and Zn+2 from 1.97+0.05 to 1.31±0.06 (15% and 33% glucose (5.6 mmol/L, 25mmol/L and 33.6mmol/L) and harvested at the indi- respectively). Group B: BG rose from 2.9± 0.2 mM to 4.6±0.5 mM; PS and cated time (12h and 24h). After treated with hyperglycemia for 12h and 24h, Zn+2 were at 1.01±0.02 and 1.04±0.03. Group C: BG constant at 4.1±0.6 mM; the apoptosis rate was obviously higher in 25mmol/L and 33.6mmol/L group histology unchanged; B-cell IN (PS and IG) achieved 1.75± 0.09, with Zn+2 than in 5.6mmol/L group (p<0.05), while cell proliferation was signifi cantly 1.34±0.03. Thus, XA formation in intact islets resulted in ~2-fold reduction in lower in 25mmol/L and 33.6mmol/L group than that in 5.6mmol/l group Zn+2 content compared with IN reduction. In absence of XA, Zn+2 content in (p<0.05); the mRNA level of CHOP, caspase-3, caspase-12 mRNA and the B-cells was restored at slower rate than IN content. We hypothesize that not protein level of CHOP were increased in a dose-dependent manner (p<0.05); all Zn+2 ions in B-cells are in the form of the IN/Zn+2 complex.

the mRNA and protein level of GRP78 were increased after treated with Islet Biology/ hyperglycemia for 12h, and then decreased at 24h (p<0.05). Hyperglycemia Insulin Secretion signifi cantly promotes MS-1 cells apoptosis and decreases proliferation PUBLISHED ONLY through endoplasmic reticulum stress pathway. And regulation of endo-

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A719