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Home , Adiponectin, Resistin

Abstracts S43 Poster session: Track 1, Molecular biology and basic physiology

T1:P1-001 T1:P1-002 Effect of chronic free estrone treatment on body weight Short-term modulation by oleoyl-estrone of plasma and composition of normal-weight Wistar in normal-weight Wistar rats Ferrer R, García B, Gómez S, Fernández-López JA, Salas A, Finazzi C, Fernández-López JA, Remesar X, Alemany M, Remesar X Esteve M, Alemany M University of Barcelona. Barcelona, Spain University of Barcelona. Barcelona, Spain

Aims: To determine whether free estrone enhances the Aims: The purpose of this study was to ascertain whether deposition of in a dose-dependent way oleoil-estrone (OE) effects on energy homoeostasis were Methods: Two-month old Wistar male and female rats were only long-term or there was a short-term component given a daily oral gavage of 0, 0.01, 0.05, 0.1, 1 or 10 nmol/g Methods: Three-month old Wistar female rats were given a estrone during 10 d. BW was measured daily, on sacrifice (d single oral gavage of 10 nmol/g of OE in 0.2 ml oil. Rats were 10); body composition was determined. killed at 3, 6, 24 and 48 h and compared with controls (C). Results: BW increase peaked at the 0.005 nmol/g dose, and BW, food intake and plasma parameters were measured decreased maximally at 10 nmol/g, the effect being more Results: Food intake and body weight decreased in OE vs. marked in males. Highest % lipid was found with the lowest C. Plasma levels were slightly lower in OE, doses of estrone, but was higher in all groups than in was also lower in OE only 6 h after gavage. Plasma TAG did controls, reversing the trend of % protein. Body cholesterol not change with OE, but both NEFA and 3-hydroxybutyrate decreased in all groups compared with controls, except at (3HB) levels increased during treatment, the effects being 0.1 nmol/g. Plasma glucose did not change, but insulin and apparent already at 6 h. Total plasma cholesterol and HDL- levels followed the same pattern than BW. cholesterol fractions decreased dramatically with OE and Conclusions: The effects of oral free estrone were biphasic: remained low from 6 h onwards. at low (i.e. physiological) doses, an enhancement in growth, Conclusions: The effects of oral OE were not yet detected 3 lipid deposition and circulating insulin and leptin were h after gavage, but at 6 h the effects on plasma insulin, observed, whilst at higher (i.e. pharmacological) doses, BW glucose, cholesterol NEFA and 3HB were already observed. insulin and leptin decreased. However, different intensities of These actions of OE were independent of food intake and decrease in plasma cholesterol and body cholesterol suggest that lipid mobilisation and handling of circulating accumulation were observed at all doses of estrone tested. lipids by OE are early events after its administration.

T1:P1-003 T1:P1-004 Oleoyl-estrone decreases plasma insulin, leptin and Chocolate Ensure and diet induced in the rat , lowering muscle and BAT glucose uptake Archer ZA, Rayner DV and Mercer JG. Grasa MM, Díaz-Silva M, Fernández-López JA, Remesar X, Rowett Research Institute, Aberdeen, Scotland, UK Alemany M. University of Barcelona. Barcelona, Spain Aims: To examine the physiological responses of feeding western style (high energy, highly palatable) diets to a Aims: To assess the insulin sensitivity of Wistar rats polygenic rat model of obesity. chronically treated with oleoyl-estrone (OE), which maintain Methods: 80 male out-bred Sprague Dawley rats weighing glycaemia with decreased insulinaemia. ~193g were fed chow (C) for 1wk. Rats were then fed high Methods: Three-month old Wistar female rats were given oral energy (HE) diet (C11024; Research diets) for 3wks and rats gavages of 10 nmol/g of OE in 0.2 ml oil (OE group), or only divided into 2 weight-matched groups: for 10wks half oil (C). After 8 days, insulin, leptin, adiponectin and glucose remained on HE and half had access to chocolate Ensure were measured in plasma; oral glucose tolerance tests (EN; highly palatable liquid diet) in addition to HE. 20 rats [OGTT] and hyperinsulinaemic-euglycaemic clamps were from the HE and EN groups were then killed and tissues performed in anaesthetised C and OE rats. collected. The remaining rats were returned to C for 3wks Results: OE rats lowered plasma insulin, adiponectin, and and then killed. Bodyweight and food intake were measured. leptin, maintaining glycaemia. In OE, OGTT resulted in lower Results: Irrespective of diet, rats exhibited a range of weight area under the curve [AuC] for insulin and no changes in gains. Compared to HE, EN rats had an increased weight AuC for glucose. However, OE rats showed a lower rate of gain with a differential of 78g by wk10. Energy intake was glucose infusion during the clamp, with decreased 2- higher in EN than HE rats throughout the 10wks. On transfer deoxyglucose transport into BAT and as well as in back to C, energy intake and bodyweight decreased in EN soleus and extensor digitorum longus muscles. rats, whereas HE rats maintained their bodyweight. Conclusions: OE increased insulin sensitivity, as shown by Conclusions: Physiological responses to dietary lower basal insulinaemia and the OGTT data. Muscle and manipulations differentially affect the level of bodyweight that BAT decreases in glucose uptake in OE rats agree with the will be defended. The underlying mechanisms involved are lower insulin and adiponectin levels, sparing glucose and likely to be important in the development of human obesity. thus favouring the utilization of lipid by muscle for energy. Funded by EU (QLK1-2000-00515).

International Journal of Obesity Abstracts S44 T1:P1-005 T1:P1-006 Effect of leptin on in ob/ob mice Evaluation of alpha concentration Asensio C, Rohner-Jeanrenaud F, Muzzin P (TNF) in patients with syndrome Département de Biochimie Médicale, Centre Médical Bogdanski P., Pupek-Musialik D., Cymerys M., Luczak M., Universitaire and Division d’Endocrinologie et de Bryl W., Szulinska M. Diabétologie, HUG, Geneva, Switzerland Department of Internal and Metabolic Disorders, University of Medical Sciences, Poznan, Poland Aims: Resistin is a expressed and secreted by white (WAT). It has been proposed to be a Aim of the study was evaluation of TNF concentration in patients with clinical features of insulin resistance (IR) and investigation of factor that contributes to insulin resistance in obesity. Based the relationship between TNF and selected anthropometric and on the observation that leptin administration ameliorates biochemical parameters. insulin resistance of obese mice lacking leptin (ob/ob mice), Methods: 34 patients with and hypertension we hypothesized that leptin may modulate resistin (n=34, aged 47.3±13.2; BMI=38.6±6.1kg/m2) were analysed. 16 expression. healthy lean volunteers constituted a control. Dependent (%FAT) Methods: We determined resistin and protein fat content was measured by bioimpedancy method. TNF and expression in WAT of ob/ob mice and tested the effects of 4 insulin were assessed using RIA method. Insulin resistance was days intraperitoneal leptin injections in these mice. calculated as fasting insulin/glucose ratio (IRI/G). Results: We report here that both resistin mRNA and protein Results: levels of TNF in the study group exceeded those levels are lower in WAT of ob/ob mice than in that of their observed in a control (P <0.05). Higher values of insulin and lean littermates and that the leptin treatment further down- IRI/G ratio in this group were observed (P<0.05). Positive regulated resistin gene and protein expression in ob/ob mice. correlations between TNF and %FAT (r=0.58; P<0.05) and Conclusions: The downregulation of resistin by leptin in WAT between TNF and IRI/G ratio (r=0.69; P<0.05) were found. of ob/ob mice is fitting with the known insulin resistance- Conclusions: 1. Patients with clinical features of IR have ameliorating effects of leptin. We are currently testing the increased level of TNF. 2. Positive correlation between %FAT effect of resistin in insulin-sensitive tissues. The results of and TNF can indicate adipose tissue as an important source of this study will be presented at the congress. this . 3. Positive correlation between TNF and IRI/G ratio suggests that TNF should be considered as an important player in the state of insulin resistance.

T1:P1-007 T1:P1-008 Modeling of resting energy expenditure from organ- The effect of vitamin A on energy and the tissue masses in underweight and obese adults expression of UCP2, FAS and delta-6-desaturase Bosy-Westphal A, Reinecke U, Schlörke T, Illner K, Braunerova R, Hainer V, Kunesova M, Parizkova J, Wagenknecht M, Sponarova J, Kopecky J. Kutzner D, Heller M, Müller MJ rd st Institut of Human Nutrition, University Kiel, Kiel, Germany Institute of Endocrinology and 3 Dept. of , 1 Fac. of Medicine, Prague, Czech Republic Aim: To calculate resting energy expenditure (REE) from organ-tissue masses over a wide range of fat-free mass Aims: To evaluate whether the increase of vitamin A (FFM) applying specific organ metabolic rates. consumption influences resting metabolic rate (RMR), Methods: Cross-sectional study on 57 healthy adults (35 substrate oxidation at rest (defined as RQ) and the females and 22 males, 19-43yr; 14 underweight, 25 normal expression of UCP2, FAS and delta-6-desaturase genes in weight and 18 obese). REE was measured by indirect obese women characterised by low intake of vitamin A. calorimetry (REEm) and calculated (REEc) from masses of Methods: 9 obese women with decreased intake of vitamin A , internal organs, (MM), bone and (<50% RDA) were administered 30 000 IU/d of retinol for 4 adipose tissue (assessed by MRI and DXA) assuming weeks. RMR and fasting non-protein RQ were determined by constant tissue respiration rates. indirect calorimetry. Expression of UCP2, FAS and Results: REEm and REEc showed a high agreement over delta-6-desaturase mRNA in subcutaneous white adipose the FFM range (28-86kg). REE prediction errors were –17 tissue was assessed by real time RT-PCR. ±505, –145 ±514 and –141 ±1058 kJ/d in normal weight, Results: RMR and UCP2 expression tended to increase and underweight and obese subjects, respectively (n.s.). FAS and delta-6-desaturase expression tended to decrease. Regressing REEm on FFM resulted in a positive intercept No significant effects of increased vitamin A consumption on of 1.6 MJ/d that could be reduced to 0.5 MJ/d by adjusting RMR, RQ and the expression of the above-mentioned genes FFM for the proportion of MM/organ mass. 81% of the were found. variance in REEm was explained by MM and mass. Conclusion: The increased intake of vitamin A did not Conclusion: Detailed analysis of metabolically active influence RMR, RQ or UCP2, FAS and delta-6-desaturase compartments of FFM allows modeling of REE over a wide expression in obese women. range of FFM. Non-linearity of REE on FFM could be partly Supported by grant COST B17-40 and IGA 7031-3. explained by FFM-composition.

International Journal of Obesity Abstracts S45 T1:P1-009 T1:P1-010 Regulation of Adiponectin by Adipose Tissue-derived Low-grade inflammatory state associated to obesity is related to the adipose tissue TNFα and leptin expression. Jens M. Bruun, Aina S. Lihn, Camilla Verdich, Steen B. Bulló M, García-Lorda P, Megias I, Salas-Salvadó J. Pedersen, Søren Toubro, Arne Astrup and Bjørn Richelsen. Human Nutrition Unit, Faculty of Medicine. Universitat Rovira i Department of Endocrinology and Metabolism, Aarhus Virgili. Reus, Spain. Amtssygehus, DK-8000 Aarhus C, Denmark Aims: To explore the links between TNFα and leptin adipose Aims: To investigate the interaction between adiponectin and tissue expression, and systemic (CRP and IL-6) adipose tissue-derived cytokines: interleukin-6 (IL-6), and the relationship between inflammation, adiposity, type 2 interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). and other cardiovascular risk factors. 2 Methods: 1) Adiponectin levels in plasma and adipose tissue- Methods: 91 women (BMI 19-65 kg/m ) were divided into biopsies from obese subjects before and after weigh t loss. 2) CRP tertiles. Plasma glucose, lipid profile, IL-6, sTNFR1 and The direct effect of IL-6, IL-8 and TNF- α on the expression 2, leptin and serum CRP levels were measured. Adipose α and release of adiponectin in whole adipose tissue cultures. tissue mRNA of TNF and leptin was also measured. Results: Weight loss resulted in a 51 % (p<0.05) increase Results: CRP and IL-6 were positively related to BMI and waist-to-hip ratio. CRP was related with HOMA IR, in plasma adiponectin levels, parallelled by a significant increase , VLDL-cholesterol, sTNFR1, sTNFR2, leptin, in adiponectin levels in the adipose tissue. Plasma adiponectin TNFα mRNA and leptin mRNA (p<0.05). Plasma sTNFRs was inversely correlated with adiposity, insulin sensitivity and the and leptin, and TNF and leptin mRNA were higher in patients level of cytokines investigated, particular IL-6. In vitro adiponectin with the greatest inflammation grade. BMI, was decreased when incubating with TNF- α and TNFα mRNA levels were significant predictors of serum (p < 0.001) and IL-6 (p < 0.01). CRP levels (r2=0.28, p<0.001). Conclusions: The inverse relationship between adiponectin Conclusions: These results are in agreement with the and cytokines in vivo and the cytokine-induced reduction in hypothesis that the synthesis of adipose tissue TNFα and adiponectin mRNA level in vitro suggests that endogenous leptin could induce the production of IL-6, CRP and other cytokines may inhibit adiponectin. acute phase reactants, thus contributing to the maintenance of chronic low-grade inflammation state involved in the progression of obesity and its associated comorbidities.

T1:P1-011 T1:P1-012 WEEKEND EXERCISE PROMOTING MODIFICATIONS ON Plasma leptin and insulin levels on 5 years body weight LEPTIN CONCENTRATION AND LIPOGENISIS IN and BMI changes among school children in Taiwan --- HYPERCHOLESTEROLEMIC MALE ADULT RATS. the Taipei Children Heart Study N.C. Cheik; R. L. F. Guerra; L. M. Oyama; C.M.O Nascimento; Chu NF, Shen MH, Wu DM E. A. Rossi; I.Z. Carlos; K.O. Nonaka; A. R. Dâmaso. Department of Community Medicine and Public Health, DEF/PPGCF/CCBS/UFSCAR; FCF-UNESP Araraquara; Tri-Service General Hospital, NDMC, Taipei, Taiwan, ROC UNIFESP-EPM - São Paulo, Brazil. Can weekend exercise be effective to change Leptin concentration and Aims: To evaluate the association of plasma leptin and lipogenesis? OBJECTIVE: To show if weekend exercise can be insulin levels on 5 years body weight (BW) and body mass effective to change Leptin concentration and lipogenesis in Wistar male adults rats. METHODS: Wistar male adults rats were divided in six index (BMI) changes among school children. groups: Normocaloric and Hypercholesterolemic: Sedentary (NS; HS), Methods: The Taipei Children Heart Study was a longitudinal Continuous Training(CTN; CTH, 5days/week/90minutes/day), study from 1995 to 2000 to ascertain anthropometric and Intermittent Training(ITN; ITH, 2days/week/90minutes /day). After 08 cardiovascular risk factor among children in Taiwan. We weeks of training and treatment, the animals were killed. enrolled 718 children (376 boys and 342 girls), aged 13 to 15 Retroperitoneal and epididimal white adipose tissues and brown years, to collect their BW and BMI changes during 5 years adipose tissue were collected for lipogenesis measurements (L) and period. We also measured baseline (in 1995) plasma leptin was collected for leptin analyze. RESULTS: and insulin levels using RIA method. G/T NS CTN ITN HS CTH ITH Results: Overweight children had higher plasma leptin and Leptin 3,12 ± 1,87 ± 2,34 ± 2,19 ± 1,64 ± 2,11 ± 1,04 0,53∗ 0,47 0,89 0,22 0,54 insulin levels than normal weight at baseline. Plasma leptin L. RET 1,34 ± 0,47 ± 0,17 ± 1,26 ± 0,82 ± 0,82 ± 0,29 0,03∗ 0,02∗ 0,11 0,10 0,18 and insulin levels were positively correlated with baseline L. EPI 0,59 ± 0,32 ± 0,33 ± 1,30 ± 0,68 ± 0,59 ± 0,11 0,04 0,04 0,20 0,06∗ 0,07∗ and 5-year followed BW and BMI. Plasma leptin levels were L. BAT 0,99 ± 1,74 ± 2,32 ± 6,54 ± 7,12 ± 8,64 ± negatively associated with 5-year BMI changes (r=-0.36 for 0,22 0,39 0,23 0,48 0,60 0,90 boys and –0.41 for girls, all p <0.001). These associations ≤ * p 0.05 exercise vs. sedentary were persisted even after adjusting for insulin level and other CONCLUSION:The weekend exercise was able to promote effects in the leptin concentration and lipogenesis rate in different adipose potential confounders. tissues, resulting on control of the obesity in Hypercholesterolemic rats. Conclusions: Plasma leptin may play a significantly role on 5-year body weight and BMI changes among school children in Taiwan.

International Journal of Obesity Abstracts S46 T1:P1-013 T1:P1-014 Sodium Tungstate Modulates PPARγ Function To Exert Its Impaired glucose tolerance (IGT): skeletal muscle fatty Antiobesity Effect acid handling after a high fat meal Corominola H., Claret M., Canals N., Barceló S. and Gomis R. Corpeleijn E, Mensink M, Saris WHM, Blaak EE, Endocrinology and Diabetes Unit, IDIBAPS, Hospital Clinic, University of Maastricht, Maastricht, The Netherlands Barcelona, Spain Aims: Do obese IGT men have an impaired muscle Our previous studies showed that oral administration of sodium uptake and/or oxidation in response to a high fat meal tungstate restores insulin sensitivity and decreases body weight compared to obese controls? in an experimental model of diet-induced obesity. Aims: To Methods: 7 IGT and 6 control (C) men with comparable BMI determine whether tungstate modulates: 1) (IGT= 32.4±3.3, C= 36.4±4.9 kg/m2), bicycle Vo max (IGT= differentiation/apoptotic processes, 2) PPARγ -regulated gene 2 41.6±5.1, C= 37.7±5.4 ml O /kg FFM) and age (IGT= 52± 6, transcription and 3) p42/p44 MAPK pathway to regulate PPARγ 2 C= 58±5 y.) participated in the study. Skeletal muscle activity. Methods: Epidydimal white adipose tissue (eWAT) sections from obese treated (2mg/ml) and untreated rats were substrate metabolism was studied during baseline (B) and stained with haematoxylin/eosin. Apoptosis was detected by the postprandial (PP) conditions (for 4 hrs after a meal: 60 en% TUNEL technique. Pooled cDNA was used for semiquantitative fat, 30 en% CHO, 10 en% protein) by means of the forearm PCR amplification with specific gene primers for: UCP1, ACO, balance technique and muscle local indirect calorimetry. FAS/CD36, HSL, m-CPT1 and LPL. MAPK activation was Results: No differences were seen in blood flow, arterial free assessed by Western blot. Results: Tungstate treatment induced glycerol, free fatty acid and concentrations both in apoptosis in eWAT from obese treated vs obese untreated rats the basal as in the postprandial state. PP arterial glucose (7.8+2.7% vs 3.2+1.2%, p<0.05). It also caused a shift in the cell concentrations increased more in IGT men (p<0.05) than in population that results in small newly differentiate . controls. Skeletal muscle fluxes in glucose, lactate, TAG, free Preliminary results showed decreased expression of genes glycerol and FFA (B: IGT=-39.2±105.3, C=15.0±78.8; PP: regulated by PPARγ and a strong activation of MAPK in obese IGT= -9.4±63.6, C=0.7±63.7 nmol/100ml tissue/min) were not treated rats. Ectopic expression of UCP1 was not observed. different, nor was muscle respiratory quotient. Conclusions: These results suggest that repression of PPARγ Conclusions: These preliminary data show no differences in activity via a MAPK-dependent pathway might be the mechanism skeletal muscle FFA handling during postprandial conditions through which tungstate exerts its antiobesity effects. This could in IGT subjects. Stable isotope tracer data on FFA uptake be a novel therapeutical approach for the treatment of obesity. and release will be available by the time of the congress.

T1:P1-015 T1:P1-016 METABOLIC EFFECTS OF BIOCATALYTIC OXYGENATION. and related factors in De Cristofaro P, Campanaro P, Dragani B, Malatesta G, Drevon CA, Lips E, Haugen F, Andersen MH, Harsem NK, Battistini NC* and Pietrobelli A**. Staff A & Ranheim T. Centro Regionale di Fisiopatologia della Nutrizione ASL Inst Nutrition Research & Dept Mother & Child Ullevaal Teramo, *Cattedra di Nutrizione Umana, Università di Modena, Hospital, University of Oslo, Norway **Clinica Pediatria, Università di Verona (Italia). Aim: We tested the biocatalytic oxygenation (BO) method Aims: Examine expression of some adipokines and related consisting in respiratory integration with therpenic compounds factors in placenta and plasma of normal and preeclamptic alpha and beta pinene derived from Pinus pinaster resins women. peroxidized by technological procedure (Holiste, France). Methods: Fasting (> 6 h) blood samples were taken before Methods: 80 females were randomly divided into two groups cesarean section and placental tissue was collected just after (A and B), and then evaluated by indirect calorimeter. Group A birth. Northern blotting, PCR, RIA and ELISA were used. performed 6 minutes of BO, whereas Group B performed 6 Results: Resistin mRNA in placenta was similar in normal minutes of free respiration. Both groups were followed by and preeclamptic women with a 3.5-fold increase in leptin indirect calorimeter as control. γ Results: Group A showed significant reduction of respiratory mRNA in preeclampsia. PPAR and SREBP-1 mRNA were quotient (QR) (p=0.05) after BO. Patients with BMI (kg/m2) reduced by 20 and 30 %, while C/EBPα was similar in both between 18.5 to 29.9 were selected from group A and B and groups. Fatty acid synthase mRNA was reduced by 33 % in divided in two subgroups (A1 and B1) of 21 and 22 patients preeclampsia, while LDLreceptor mRNA was unaltered. respectively, homogeneous for age, and anthropometry. Plasma concentration of IL-6, TNFα, leptin and adiponectin Group A1, showed significant increase in VO2 (p=0.01), high were 2-fold increased in preeclampsia. Cultures of human significant reduction of QR (p=0.0001), and high correlation placental syncytotphoblasts exhibited low resistin mRNA. between ∆QR and ∆CO2 (r=0.77; p<0.0001) after BO. Conclusions: Placental resistin mRNA is similarly expressed Conclusions: Increase of tissue oxygen biodisponibility and in placenta from preeclamptic and normal women. mRNA of improvement of lipids/carbohydrates oxidation ratio in the PPARγ and SREBP-1 is decreased in placenta from group performed BO underlined the correlation between preeclamptic women, whereas plasma levels of leptin, respiration and substrates-oxidation. Our findings may suggest adiponectin, IL-6 and TNF-α are elevated. a therapeutic utilization of BO.

International Journal of Obesity Abstracts S47 T1:P1-017 T1:P1-018 Postprandial hyperlipidemia: Effect of acute exercise Gene Expression Profiling in Human Adipose Tissue L.H.Enevoldsen, L.Simonsen and J.Bülow Engeli S, Baranov S, Soldatov A, Lehrach H, Gorzelniak K, Department of Clinical Physiology, Bispebjerg University Heintze U, Janke J, Luft FC, Sharma AM, Franz Volhard Hospital, DK-2400 Copenhagen NV, Denmark Clinical Research Center, Charité, Humboldt University of Berlin, Germany Aims: Hyperlipidemia is a significant risk factor for the development of and insulin resistance. The Aims: To investigate adipose tissue gene expression profiles aim of the present study was to unravel the effect of acute under certain conditions (obesity, insulin resistance, exercise on postprandial hyperlipidemia. hypertension, weight loss, pharmacological therapy). Methods: Four healthy male subjects were studied twice. In Methods: In a cross-sectional study, subcutaneous adipose one study subjects were studied during resting conditions tissue samples were obtained from 80 drug-free menopausal before and for 4 hours after a carbohydrate rich meal. In a women and hybridized to 3300 cDNA clones previously second study the subjects exercised for 1 hour after the meal chosen due to differential expression in a subset of the study and then studied for another 3 hours. Local adipose tissue population. Intervention studies are currently running that will was determined by Fick’s Principle, and postprandial be analyzed by the meanwhile established protocols. lipidemia by integration of arterial concentrations. Results: The comparison of lean vs. obese subjects revealed Results: Compared to the resting experiment exercise a total of 250 cDNA clones differentially expressed at a p induced a significant increase in circulating TAG, but over all value < 0.05. Of these, about 130 have perfect hits to named the postprandial hyperlipidemia did not differ between the genes in available databases (eg. IGF1, IGF2, atrial two experiments. Adipose tissue lipolysis during exercise and type A of this protein). All of was similar to our previous findings in fasting subjects, while these clones are currently sequenced to confirm gene concomitantly there was a significant TAG clearance in the annotation. Following this essential step, more in depth present experiment. During exercise there was a significant analyzes will be performed to integrate gene expression data reesterification in adipose tissue. and clinical parameters (cluster analysis, and others). Conclusion: Exercise for 1 hour after a carbohydrate rich Conclusions: This large gene expression profiling study will meal does not influence the postprandial hyperlipidemia, but offer new chances to understand the contribution of adipose adipose tissue reesterification is increased during exercise. tissue to obesity-associated conditions such as hypertension.

T1:P1-019 T1:P1-020 Regulation of 11ß-Hydroxisteroid-dehydrogenase 1 and Synergic effect of wine phenolic compounds and 2 genes in human adipose tissue oleoyl-estrone on the loss of body weight in obese Gorzelniak K, Engeli S, Janke J, Feldpausch M, Heintze U, Zucker fa/fa rats. Luft FC, Sharma AM, Franz Volhard Clinical Research Romero M, Esteve M Center, Charité, Humboldt University of Berlin, Germany University of Barcelona. Barcelona, Spain

Aims: Overexpression of 11ß-Hydroxisteroid-dehydrogenase Aims: To determine a possible synergic effect between 1 (11ß-HSD1) in mice adipocytes increased local formation oleoyl-estrone (oe) and wine phenolic compounds (wpc), of and is associated with the . In since both compounds share effects on lipids handling and obese subjects, 11ß-HSD1 activity in adipose tissue is mobilisation. increased due to unknown mechanisms. We determined Methods: Three-month old obese male Zucker fa/fa rats were gene expression levels 11ß-HSD1 and 2 in adipose tissue. given daily oral gavages of oil (C), of oe 10 nmol/g·d in oil Methods: Sc adipose tissue samples were obtained from (OE ), of wpc 15 nmol/g·d in oil (PC) or both oe 10 nmol/g·d postmenopausal women, weight reduction was achieved by and wpc 15 nmol/g·d in oil (OE+PC). After 30 days of hypocaloric diet and exercise. Gene expression was treatment the rats were killed. Body weight change, food determined by TaqMan real time RT-PCR. intake and plasma parameters were measured. Results: 11ß-HSD1 expression is positively associated with Results: No differences in body weight and food intake were BMI (r2 = 0.19, p < 0.001), whereas 11ß-HSD2 gene found in C and PC groups. Decreases in body weight were expression is negatively associated with BMI (r2 = 0.11, p < higher in OE+PC than in the OE, but no differences in food 0,05). Body weight reduction of 5% in 12 weeks in a subset intake were found between both groups. Plasma glucose, of these women (n=7) did not change gene expression. insulin, NEFA, TAG and 3-hydroxybutyrate were lower in OE Conclusions: Upregulation of the 11ß-HSD1 gene and and OE+PC vs C and PC groups. Plasma leptin increased in concomittant downregulation of the 11ß-HSD2 gene may PC and decreased in the OE and PC+OE groups. Plasma contribute to increased cortisol formation in adipose tissue of acyl-estrone levels were higher in PC vs the C group. obese subjects. Weight loss of 5%, however, did not Conclusions: This effect may be mediated by a protective efficiently influence expression levels of both HSD genes. effect of the wpc on the handling and stability of oe and also by enhancing WAT ponderostat signals: oe + leptin by itself.

International Journal of Obesity Abstracts S48 T1:P1-021 T1:P1-022 DECREASED MUSCLE LIPID OXIDATIVE CAPACITY IN Retinoic acid inhibits resistin expression OBESE SUBJECTS AFTER WEIGHT LOSS. Felipe F, Bonet ML, Ribot J, Palou A. Universitat de les Illes Fabris R., Milan G., Granzotto M., Mingrone G., Manco M., Balears, Palma de Mallorca, Spain Serra R., Scarda A., Greco A.V., Federspil G., Vettor R. Dept. of Medical & Surgical Sciences, Padova, Italy. Aims: Resistin was discovered as an adipocyte gene product down-regulated by antidiabetic drugs (TZDs) and involved in Aims: A low resting energy expenditure and a reduced ability insulin resistance, although results are conflicting. TZDs are to oxidize fat are risk factors for weight gain. Obese subjects PPAR ligands and PPARs regulate transcription as show a decrease in fat oxidation, not completely defined at heterodimers with 9-cis retinoic acid -activated RXRs. Our molecular level. We investigated the expression of genes of main aim was to assess the effect of retinoic acid (RA) on oxidative and lipid synthesis pathways by RT-PCR: PPARα, resistin expression. Acyl-CoA Oxidase (ACO), Carnitine Palmitoyl Transferase 1B Methods: Resistin mRNA was measured by northern blot in (CPT1B), Acetyl-CoA Carboxylase (ACAC)β and Fatty Acid fat depots of mice acutely treated with RA or chronically fed Synthase(FAS) in skeletal muscle biopsies from obese vitamin A-supplemented diets, and in 3T3-L1 cells and patients before and after biliopancreatic diversion (BPD). primary cultures of brown adipocytes treated with retinoids. Methods: Euglycemic clamp was performed in 10 obese Results: Resistin mRNA was reduced in vivo after RA- subjects before and after weight loss. More over the rates of treatment and vitamin A supplementation. Both 9-cis RA and glucose and lipid oxidation in respiratory chamber,and muscle all-trans RA down-regulated resistin mRNA expression in triglyceride (TG) concentration were measured. vitro; the effect was time- and dose-dependent, reproduced Results: Weight reduction improved insulin sensitivity. An by both RAR and RXR and partially blocked by increase in glucose oxidation and a decline in lipid oxidation antagonists of these receptors. and muscle TG content were observed after BPD. PPARα and Conclusions: 1) RA is a direct inhibitor of resistin gene CPT1B expression was significantly reduced. expression. 2) The effect of RA appears to be mediated by Conclusions: Our results suggest the presence of a defect of both RARs and RXRs. 3) Vitamin A status affects resistin both peroxisomal and mitochondrial oxidative pathways at expression. RXR agonists are known to improve insulin muscular level that may contribute to the reduced fat oxidation sensitivity in animal models, and thus our results fit with in formerly obese subjects. resistin being involved in insulin resistance in the mouse.

T1:P1-023 T1:P1-024 Comparison of Different Rat Models for Determination of UCP3 protein and messager are differentially regulated in Drug Effects on Body Weight skeletal muscle of mouse exposed to high oxygen concentration. Fhölenhag K, Svartengren J, Axelsson-Lendin P, Arrhenius- Flandin-blety P, Donati Y, Arboit P, Kühne F, Barrazone C, Nyberg V, Larsson C, Klingström G Piguet PF, Muzzin P Biovitrum AB, Department of Biology, Stockholm, Sweden Medical School of Geneva, Geneva, Switzerland.

Aims: Studies in man and animals demonstrate that enhanced Aims: The 3 (UCP3) is a member of the serotonergic neurotransmission can decrease food intake and mitochondrial carrier protein family. It has been proposed body weight. We have compared different rat models for that high UCP3 levels could protect against reactive oxygen evaluation of food intake and body weight after administration species (ROS) toxicity. To get some evidence on its possible of mCPP and sibutramine. role in oxidative stress, we have examined the effect of Methods: Studies were performed in diet induced obese (DIO) hyperoxia on UCP3 expression. male rats, in old female obese and “resistant” lean rats and in Methods: C57/Bl6 mice were exposed to 95% O during 72 growing male rats. The substance administration was done by 2 hours and their UCP3 mRNA and protein expression were osmotic minipumps with 14 days duration. Results: mCPP is 2-3 fold more potent to reduce body weight in analyzed in skeletal muscle. DIO rats than in lean rats. The same pattern is seen with old Results: Under hyperoxia, UCP3 mRNA expression and female rats. mCPP is less effective in growing male rats as circulating free fatty acid increased respectively by 4.7- compared to DIO animals. Sibutramine and mCPP seem to be (p=0.057) and 3-fold (p=0.022) compared to control. equipotent. A strong correlation between body weight and Unexpectedly, UCP3 protein was down-regulated by 48% leptin concentration in serum is observed in both females and (p=0.03). The amount of mitochondrial protein per muscle males. The regression lines are parallel but not overlapping. was unchanged between hyperoxic and control mice. Conclusions: All the described rat models are relevant to use Conclusion: We observed a decrease of UCP3 protein that for determination of body weight reduction after changes in does not support a protective role of UCP3 against ROS. serotonergic neurotransmission. However, the effect is more These results also suggest that UCP3 expression may be pronounced in the obese animals. regulated at the translational level.

International Journal of Obesity Abstracts S49 T1:P1-025 T1:P1-026 EFFECTS OF TWO ALIMENTARY STATUS AND Leptin inhibits angiotensin II-induced vasoconstriction PHYSICAL TRAINING ON MUSCULAR via a nitricoxide dependent mechanism CHARACTERISTICS OF RATS Rodríguez A, Fortuño A, Gómez-Ambrosi J, Salvador J, Díez J, Freitas Jr. Ismael Forte; Gomes, Robinson Rodrigues; Frühbeck G Sabino , João Paulo Jacob; Dal Pai, Vitalino Metabolic Research Laboratory and Cardiovascular Pathophysiology Division, University of Navarra, Pamplona, Spain São Paulo State University – UNESP Aims: To analyse the effect of leptin on the vasoconstriction exert- Presidente Prudente, São Paulo, Brazil ed by known vasoactive agents and to determine the potential role of nitric oxide (NO) on these actions in the rat aorta. The purpose of this study was to verify the morfologic Methods: The ability of leptin to modify angiotensin II (Ang II)-, adaptations in gastrocnemius muscle of rats submitted to noradrenaline (NA)- and -1 (ET-1)-induced vasocon- two different weekly frequencies of training and two different striction on denuded aortic rings was tested by an or- alimentary diets. Methods: Thirty male Wistar rats were gan bath system. Experiments were repeated preincubating the studied and submitted to eight serial weeks of physical aortic rings with a non-selective inhibitor of NOS (L-NMMA) and a training. The animals were distributed in two groups: one selective inhibitor of iNOS (L-NIL)before leptin addition. Vascular sedentary(10 rats) and trained(20 rats). These trained were smooth muscle cells (VSMCs) were stimulated with leptin 10-8M then subdivided in continuous training(5 t/w) and sporadic during different time periods (0.5, 1, 2, 4, 6, 18 and 24h); culture training (2 t/w).Half of animals were fed with baseline diet medium was collected and nitrite/nitrate concentrations were- measured. and the other half with hipercaloric diet. At the end, Results: No effect of leptin on NA- and ET-1-induced vasocon- fragments of the gastrocnemius muscle were frozen in liquid striction was observed. Ang II 10-7M-induced vasoconstriction was nitrogen. Results:There was observed structural modifications inhibited (p<0.05) by leptin 10-8M. Leptin inhibition of the Ang II-in- by alterations in distribution and in morphology of fibers, duced contraction was significantly (p<0.05) reversed by preincu- such alterations were more significant in rats submitted to bation with both L-NMMA 10-5 M and L-NIL10-5 M. Furthermore, it continuous training and fed with hipercaloric diet. was observed that leptin 10-8 M increased (p<0.05) NO release by Conclusion: Continuous and regular physical training, promoved VSMCs 1 h after stimulation. larger morphologic adaptations than sporadic training, Conclusions: Leptin specifically inhibits Ang II-induced vasocon- especially in hipercaloric group. striction in endothelium denuded aortic rings. This effect is mediat- ed via a NO mechanism, since the inhibitory effect of leptin is blocked after NOS or iNOS activity inhibition.

T1:P1-027 T1:P1-028 Leptin inhibits angiotensin II-induced vasoconstriction via a Upper and lower body adipose tissue function: a direct nitricoxide dependent mechanism comparison of fat mobilization Rodríguez A, Fortuño A, Gómez-Ambrosi J, Salvador J, Díez J, Goossens GH, Tan GD, Humphreys SM, Vidal H, Karpe F Frühbeck G Metabolic Research Laboratory and Cardiovascular Oxford Centre for Diabetes, Endocrinology and Metabolism, Pathophysiology Division, University of Navarra, Pamplona, Spain Churchill Hospital, University of Oxford, Oxford, UK

Aims: To analyse the effect of leptin on the vasoconstriction exerted Lower body fat is not associated with the same risk of by known vasoactive agents and to determine the potential role of cardiovascular as fat in the upper body. Is this nitric oxide (NO) on these actions in the rat aorta. explained by differences in the physiological function? Methods: The ability of leptin to modify angiotensin II (Ang II)-, Aim: (1) To determine whether fat mobilization and blood noradrenaline (NA)- and endothelin-1 (ET-1)-induced flow differ between gluteal and abdominal adipose tissues in vasoconstriction on endothelium denuded aortic rings was tested by an organ bath system. Experiments were repeated preincubating humans. (2) To develop a new technique to assess gluteal the aortic rings with a non-selective inhibitor of NOS (L-NMMA) and adipose tissue function directly. a selective inhibitor of iNOS (L-NIL) before leptin addition. Vascular Methods: Functional studies of adipose tissue involved the smooth muscle cells (VSMCs) were stimulated with leptin 10-8 M assessment of fat mobilization by measurement of adipose during different time periods (0.5, 1, 2, 4, 6, 18 and 24 h); culture tissue blood flows, arterio-venous differences of metabolites medium was collected and nitrite/nitrate concentrations were across each depot and gene expression in tissue biopsies. measured. Results: (1) Gluteal adipose tissue had a 67% lower blood Results: No effect of leptin on NA- and ET-1-induced flow and 87% lower hormone sensitive lipase rate of action vasoconstriction was observed. Ang II 10-7 M-induced -8 than abdominal adipose tissue. Lipoprotein lipase rate of vasoconstriction was inhibited (p<0.05) by leptin 10 M. Leptin action and mRNA expression were not different between the inhibition of the Ang II-induced contraction was significantly (p<0.05) depots. (2) The first demonstration of a novel technique to reversed by preincubation with both L-NMMA 10-5 M and L-NIL 10-5 -8 investigate gluteal adipose tissue metabolism directly. M. Furthermore, it was observed that leptin 10 M increased (p<0.05) NO release by VSMCs 1 h after stimulation. Conclusion: Gluteal adipose tissue is metabolically ‘silent’ Conclusions: Leptin specifically inhibits Ang II-induced compared to that in the abdomen. In other words, abdominal vasoconstriction inendothelium denuded aortic rings. This effect is fat exposes the body to more adverse metabolic mediated via a NO mechanism, since the inhibitory effect of leptin is consequences than gluteal fat. blocked after NOS or iNOS activity inhibition.

International Journal of Obesity Abstracts S50 T1:P1-029 T1:P1-030 Transactivation of the leptin gene promoter by HIF-1 Cerebral activation associated with food stimuli Grosfeld A, André J, Hauguel-de Mouzon S, Berra E, Hämäläinen S, Karhunen L, Carlson S, Martinkauppi S, Pouysségur J and Guerre-Millo M. Martikainen E, Kuikka JT, Könönen M, Pietiläinen K, INSERM U 465, Paris, France. Rissanen A, Keski-Rahkonen A, Tiihonen J, Vanninen R, Vanninen E, Aronen H, Poutanen V-P, Uusitupa M, Kaprio J Aims: We have shown that hypoxia activates the human University of Kuopio, Kuopio, Finland leptin promoter in trophoblast-derived BeWo cells. Here, we tested whether the hypoxia-inducible factor 1 (HIF-1), an Aims: To evaluate cerebral regions involved in the HIF-1α/HIF-1β heterodimer that regulates the transcription of processing of visual food, non-food, unpleasant, pleasant hypoxia-responsive genes, activates the leptin promoter. and neutral stimuli. Methods: BeWo cells were transiently transfected with Methods: The different visual stimulation conditions and a luciferase constructs containing various lengths of the leptin baseline control condition were presented in a counter- promoter. To increase HIF-1 activity, the cells were either balanced manner using block design. Whole-brain fMRI was cultured with CoCl2, that inhibits O2-dependent degradation conducted on a 1,5 T MRI scanner. fMRI data analysis was of HIF-1α, or transfected with a vector encoding wild-type performed by SPM99. So far, 13 (7 female, 6 male) 24-27 yr HIF-1α. A dominant negative HIF-1α was used in some cells. old healthy monozygotic twin pairs from the FinnTwin16 Results: Exogenous HIF-1α or CoCl2 markedly activated study have been examined. leptin promoter activity when HIF-1β was co-expressed. The Results: Significant signal intensity changes were found in dominant negative HIF-1α mutant abolished activation of the the left posterior cingulate cortex (comparison food > non- promoter by either HIF-1α or CoCl2. Deletion analysis, food), bilaterally in the amygdala (unpleasant > baseline) and mutagenesis and electrophoretic mobility shift assays in the left amygdala (food > baseline). The contribution of the revealed that a HIF-1 consensus binding site (HRE), body weight, genetic and other factors to the cerebral spanning –120 to –116 bp, was required for CoCl2 and response will be analysed. exogenous HIF-1α induction of leptin promoter activity. Conclusions: These tentative findings provide functional Conclusions: Leptin is a new hypoxia-inducible gene. The evidence that the left posterior cingulate cortex and the left stimulatory effect of hypoxia on leptin relies on HIF-1 binding amygdala might be involved in cerebral processing related to to a consensus HRE located in the proximal promoter. the sight of food.

T1:P1-031 T1:P1-032 Mice with low metabolic rates are not susceptible to Plasma in morbid obesity before and after weight gain under a high fat diet. weight loss following laparoscopical adjustable C Hambly, Adams A, Fustin JM, Rance KA, Bünger L, Hill gastric banding. WG, Speakman JR School of Biological Sciences, University Hanusch-Enserer U, Cauza E, Brabant G, Rosen HR, Pacini of Aberdeen, Aberdeen, Scotland, UK G, Dunky A, Prager R, Roden M. Wilhelminenspital, Vienna, Austria Aims: Mice that have been divergently selected for high (H) or low food intake (L) were examined to determine whether Aims: To examine the effect of weight loss following gastric provision of a diet high in fat caused significant weight gain. banding (LAGB) on plasma ghrelin and leptin and their Methods: Resting metabolic rates (RMR) have been correlation with insulin resistance (IR) in morbid obesity. previously examined and individuals from the L line have Methods: 18 (15f/3m) underwent modified oral glucose significantly lower RMR than those from the H line. It was tolerance tests (OGTT) to assess IR and insulin secretion hypothesised that this strain might increase their body mass before, 6 and 12 months after LAGB. on the high fat diet, if RMR is a risk factor for obesity. Results: Patients lost 21.4 and 31.5 kg, resulting in reduction Results: This was not the case, as neither sex nor strain of of BMI from 45.27 to 37.24 and 33.56 kg/m² (all p<0.0001). mouse significantly increased their body mass in response to These changes were associated with reduction in waist the high fat diet. In comparison to the chow diet, individuals circumference (p<0.0001), blood pressure, plasma glucose, from the H line reduced their food intake significantly by 40- insulin and IR. Plasma ghrelin was comparable before and 45% while a 31-35% decrease in food intake was seen in after 6 months (234±53 and 232±53 pmol/l) but slightly individuals from the L line. The decreases in food intake increased at 12 months (261±72; p=0.04 vs. 6 months; were seen in conjunction with an increase in their digestive p=0.08 vs. baseline). Plasma leptin was lower at 6 and 12 efficiency, which enabled mice to maintain the amount of months (p<0.0005). Ghrelin neither correlated with leptin and assimilated energy obtained from each diet and explains why body fat mass nor with IR or insulin secretion. there was no weight gain in either strain. Discussion: While short term studies suggest a direct Conclusions: Both H and L strains of mice adjusted their food ghrelin-insulin interaction, long term weight loss is intake when exposed to a high fat diet to maintain energy associated with increased plasma ghrelin independently of balance. Mice with low RMR were not differentially changes of insulin action or body mass. susceptible to mass gain when exposed to high fat diets.

International Journal of Obesity Abstracts S51 T1:P1-033 T1:P1-034 Differences in plasminogen activator inhibitor-1 in Vagal Stimulation increases gastric emptying subcutaneous versus omental adipose tissue in non-obese Heath RB, Jones R, Frayn KN, Robertson MD and obese women University of Oxford, UK He G, Pedersen S. B., Bruun J. M., Lihn A. S., Jensen P. F., Richelsen B. Dept. of Endocrinology and Metabolism C, Aarhus Aims: Vagal stimulation in combination with a fat load Amtssygehus, Aarhus, Denmark causes an exaggerated lipaemic response, yet the potential effects of gastric emptying are not fully understood. We have Aims: To investigate whether there are regional differences in used modified sham feeding (MSF) to investigate the role of the level of mRNA and production of PAI-1 in adipose tissue vagal stimulation on gastric emptying of oral fat. (AT) depots (omental vs. subcutaneous (S.C) abdominal). Methods : Eight healthy subjects (4 male/4 female) Methods: AT from both depots was obtained from obese consumed a 50 g 13C octanoate labelled fat load on two (gastric banding; BMI: 46.9kg/m2) and non-obese separate occasions. On one occasion, the fat load was (gynaecological procedures; BMI: 23.9kg/m2) women. PAI-1 preceded by the MSF for 1 h. Blood, appetite and breath mRNA was measured by RT-PCR and production by ELISA. samples were analysed for 5 h postprandially. Results: In immediately frozen AT PAI-1 mRNA level was 13 similar in two depots and no difference between obese and Results: C recovery in breath was significantly higher after non-obese women was found. However, when tissue were MSF combined with oral fat than that of oral fat ingestion cultured (72h) the PAI-1 mRNA as well as PAI-1 production alone (P<0.001). MSF in addition to oral fat elevated plasma were higher in omental as compared with S.C. AT (p<0.05). In triacylglycerol (TG) (P<0.001) as well as increasing the the culture system, the production of PAI-1 in obese subjects suppression of ghrelin, a marker of food intake (P<0.001). was higher than that in non-obese subjects in both S.C. Appetite scores showed that subjects were also significantly (p<0.05) and omental AT (p=0.19). more fuller and satisfied after the MSF (P<0.001). Conclusion: No differences in PAI-1 expression in two AT Conclusions : Vagal stimulation increased gastric emptying depots were observed in biopsies frozen immediately after of a fat load and suppressed appetite. The cephalic removal, but after incubation of the AT higher levels of PAI-1 regulation of gastric emptying and appetite will require further were found in omental AT compared with S.C. AT. Finally, the investigation. PAI-1 production in AT from obese women was higher than that of non-obese women. Thus, PAI-1 expression can be more activated in omental as compared with S.C. AT.

T1:P1-035 T1:P1-036

Heterogeneous energetic efficiency in skeletal muscle β 3-adrenoceptor knockout in C57BL/6J mice on a high mitochondrial subpopulations. fat diet depresses the insulin response to glucose Iossa S., Mollica M.P., Lionetti L., Crescenzo R., Botta M., M. Jimenez, N. Giovannini, L. Lehr, J.-P. Giacobino, J. Barletta A., Liverini G. Seydoux, F. Assimacopoulos, F. Preitner, P. Muzzin. University “Federico II”, Napoli, Italy. Département de Biochimie Médicale, Faculté de Médecine, Geneva, Switzerland Aims: To establish possible differences in energetic coupling in skeletal muscle mitochondrial subpopulations, Aims: Treatment with a selective β3-adrenergic which differ in location and function. increases insulin secretion. It has been postulated that this Methods: Subsarcolemmal (SS) and intermyofibrillar (IMF) effect is indirect and mediated by an insulinotropic factor mitochondria were isolated from hindleg skeletal muscles of secreted by the white adipose tissue. fed or 24-hour fasted rats. Titration of state 4 respiration Methods: To test this hypothesis, we studied glucose/insulin rate as a function of mitochondrial membrane potential was in β3-adrenoceptor knockout (β3KO) mice on a carried out as an indirect measurement of mitochondrial pure genetic C57BL/6J background under basal conditions proton leak. Measurements were done in the presence of and on a high fat diet for a prolonged period of time. 0.1% bovine serum albumin (basal proton leak) and 0.1% Results: β3KO mice fed 10 months a high fat diet (20% bovine serum albumin + palmitate (fatty acid induced leak). bacon fat) were glucose intolerant as compared to WT mice, Results: SS mitochondria have lower basal proton leak but without any changes in insulin sensitivity. Fasting glycemia higher fatty acid induced leak, compared to IMF was normal, whereas insulinemia was decreased 2.8-fold in mitochondria. In addition, SS mitochondria from fasted rats β3KO as compared to WT mice. Insulin peak values after an exhibit higher basal and fatty acid induced uncoupling, intraperitoneal load of glucose were a 2.6-fold lower in β3KO while energetic efficiency of IMF ones is not affected. as compared to WT mice. Conclusions: Energetic efficiency of skeletal muscle Conclusions: Our results show that a lack of β3-adrenoceptor mitochondria depends on their localisation and can be decreases insulin secretion in basal state and in response to differentially regulated by physiological stimuli. In particular, glucose. Analysis are currently performed in an attempt to SS mitochondria seem to be the population more prone to identify the β3-dependent insulinotropic factor. uncoupling, and therefore appear an attractive therapeutic target for obesity treatment.

International Journal of Obesity Abstracts S52 T1:P1-037 T1:P1-038 Differential response to interstitial angiotensin II in Nitric Oxide Release and Tissue Metabolism in Obesity normal weight and obese men Jordan J, Adams F, Klaus S, Tank J, Sharma AM, Luft FC, Boschmann M, Adams F, Klaus S, Sharma AM, Luft FC, Boschmann M. Franz-Volhard Clinical Research Center, Jordan J. Franz-Volhard Clinical Research Center, Humboldt University, Berlin, Germany. Humboldt University, Berlin, Germany Aims: The adipose tissue expression of endothelial nitric Aims: Increased expression of the tissue angiotensin system oxide synthase (eNOS) is increased in obese subjects. We has been described in obese individuals. We tested the determined the functional relevance of this increase. hypothesis that the increase is associated with increased Methods: In 8 healthy nonobese and in 8 obese subjects, we tissue responsiveness to Angiotensin II (AngII). inserted two microdialysis catheters each in abdominal Methods: In 8 healthy nonobese and in 8 obese subjects, we subcutaneous adipose tissue and in the vastus lateralis inserted microdialysis catheters in abdominal subcutaneous muscle. The catheters were perfused with L-NAME (100 µM) adipose tissue and in the vastus lateralis muscle.Tissues or D-NAME. incremental doses of Ang II (0.01, 0.1, and 1 µmol/l). Results: With D-NAME, adipose dialysate glycerol was 66± Results: During perfusion with Ang II, ethanol ratio and 7 µM in obese and 100 ± 11 µM in normal weight subjects (p dialysate glucose did not change substantially in both tissues = 0.002). In skeletal muscle, obese subjects featured an of both groups. Dialysate glycerol increased (+30%) and increase in dialysate glycerol. In obese subjects, dialysate decreased (-35%) significantly in adipose tissue and muscle, glucose and lactate were markedly reduced in adipose tissue respectively, of non-obese but not of obese men.In both but not in skeletal muscle. Tissue perfusion was unchanged groups, dialysate pyruvate was dose-dependently increased with L-NAME. The differences in tissue metabolism between (p<0.0001), where as lactate / pyruvate ratio was decreased groups were not abolished with L-NAME. (p<0.0025). Conclusions: The alterations in lipolytic activity and Conclusions: In skeletal muscle Ang II acutely inhibits carbohydrate metabolism in obese subjects are not lipolysis and glucose uptake. The effect might contribute to abolished with acute NOS inhibition. At least under basal insulin resistance. Contrary to our initial hypothesis, Ang II conditions, the increase in adipose tissue eNOS expression sensitivity was not increased in obese individuals. in obese individuals may be of limited functional relevance.

T1:P1-039 T1:P1-040 Effects of rosiglitazone on adiponectin and free fatty DIACYLGLYCEROL AFFECTSFAT OXIDATION AND acid levels in patients with type 2 diabetes mellitus APPETITE IN HUMANS Kaltsas T, Samara M, Koliakos G, Efthimiou I Kamphuis MMJW, Mela DJ*, Westerterp-Plantenga MS. Dep. of Endocrinology,Panagia Hospital,Thessaloniki,Greece Dept. Human Biology, Maastricht University, PO-box 616, 6200 MD Maastricht, The Netherlands. Aims: The aim of this study was to evaluate the effects of rosiglitazone(RSG) on adiponectin(AD) and free fatty acid(FFA) Aims: Assess effects of partial replacement of triacylglycerol levels in patients with type 2 diabetes mellitus (T2DM). (TG) by diacylglycerol (DG) on substrate oxidation, energy Methods: 15 patients with T2DM (age 62.4+/-9.4 years, expenditure (EE), blood parameters and appetite measures. diabetes duration 9.0+/-5.5 years,HbA1C 9.4+/-1.3%,BMI Methods: 12 healthy, dietary unrestrained females 31.1+/- 3.5) were enrolled in the study.We measured fasting participated in a single blind, placebo-controlled cross-over glycose,HbA1C, insulin,AD and FFA levels, as well as all design. For 3 days prior to and throughout a 36 h stay in the biochemical markers before and after 12 weeks of treatment respiration chamber, subjects ate an energy balance diet of with RSG(4 mgr od).We also estimated the degree of insulin 55/15/30 energy% as carbohydrate/protein/fat. In the resistance evaluated by homeostasis assesment models of respiration chamber, 40% of fat was consumed as DG or TG insulin resistance (HOMA-IR) oil with similar fatty acid profile. Results: Fat oxidation was Results: Compared with baseline RSG reduced fasting higher and RQ lower with DG vs. TG. Appetite profile on day glycemia(214.5+/-28.8 vs 162.6+/-29.9mg%,p<0.0005),HbA1C 1 did not differ between DG and TG, but feelings of hunger, (9,4+/-1.3 vs 7.9+/-1.0 %, p<0.005), insulin levels (15.1+/-7.1 prospective food intake, appetite and desire to eat were all vs 10.6+/-5.0 mIU/l, p<0.0005), FFA levels (48.3+/-18.8 vs lower with DG on day 2. Plasma -hydroxy-butyrate tended 35.0+/-14.5 mM, p<0.05) and HOMA-IR (142.9+/-67.3 vs to be higher with DG, and was significant at 11.30 on day 2. 74.0+/-28.6,p=0.0001) Adiponectin levels were increased after DG did not affect plasma lipid levels. There were no treatment(18.8+/-8.0 vs 29.3+/-11.9 ng/ml,p=0.0054)as well as differences in EE between DG and TG. Conclusions: Partial BMI(31.1+/-3.5 vs 32.0+/-3.7,p<0.01).Overall RSG was well replacement of TG by DG does not alter total energy tolerated with no adverse events. Conclusions: RSG increases AD and decreases FFA levels in expenditure, but produces metabolic effects, particularly T2DM patients by improving glycemic control and/or insulin increases in fat oxidation, which may be associated with resistance.This effect might prevent atherosclerotic vascular improved appetite control and energy balance. disease in insulin-resistant patients with T2DM. *Unilever Health Institute, Vlaardingen

International Journal of Obesity Abstracts S53 T1:P1-041 T1:P1-042 Serotonin transporter binding in Finnish monozygotic Effect of Obesity on the Receptor and twin pairs Insulin-like Growth Factor-I mRNA expression in Human Karhunen L, Hämäläinen S, Kuikka JT, Vanninen E, Liver and Adipose Tissues Pietiläinen K, Rissanen A, Keski-Rahkonen A, Tiihonen J, Uusitupa M, Kaprio J. SY NAM, Kim KW, H Zemac, S Norgen, C Marcus University of Kuopio, Kuopio, Finland Yonsei University, Korea, Huddinge University Hospital, Sweden

Aims: To examine SERT binding in Finnish monozygotic (MZ) Aims: We investigated the effect of obesity, in the presence of low twin pairs discordant or concordant for body weight. GH secretion on GHR and IGF-I axis. Methods: The full-length GHR (GHRfl), truncated GHR (GHRtr) Methods: So far, 13 (7 female, 6 male, age 24-27 yrs) and IGF-I mRNA expression in human liver, subcutaneous and healthy MZ twin pairs from the FinnTwin16 study have been 123 visceral adipose tissue were determined by using RNase examined. A dose of 185 MBq of [ I]nor-β-CIT (a protection assay in six obese and age-matched lean subjects. radioligand specific for SERT) was injected, and d ynamic Results: The liver GHRfl and GHRtr mRNA expressions in obese single-photon emission tomography (SPET) scans were subjects were comparable to lean subjects, whereas the IGF-I performed 5 min, 6 h and 24 h after the injection. SERT mRNA expression showed a decreasing tendency in obese binding in the midbrain was calculated according to a Logan- subjects. In visceral and subcutaneous adipose tissue both GHRfl and IGF-I expression decreased significantly in obese subjects Patlak plotas (midbrain–cerebellum)/cerebellum. compared to those in lean subjects. A strong positive correlation Results: Intrapair differences in SERT binding were between serum IGF-I level and the IGF-I mRNA expression in both significant (p=0.03) in all pairs. Individual data showed that visceral and subcutaneous adipose tissue was observed only in body weight (r=0.49, p=0.02) and fat-free mass (FFM) the obese subjects. (r=0.62, p=0.002) correlated positively with SERT binding, Conclusion: The effect of GH on GHR/IGF-I axis may be tissue FFM explaining 36% of this variation. However, intrapair specific. Although the GH hyposecretion in obesity decreases differences in weight or FFM did not explain the observed GHR/IGF-I gene expression in each adipocyte, a total amount of differences in SERT binding. GHR and IGF-I gene expression in an excessively accumulated Conclusions: Body weight and especially FFM might be adipose tissue in obesity may in part contribute to a circulating involved in the regulation of SERT binding.The possible GHBP and IGF-I level in obesity. contribution of heredity needs to be examined further.

T1:P1-043 T1:P1-044 Modulation of and lipolysis by green tea The independent and combined effects of two specialty in 3T3-L1 adipocytes carbohydrates consumed as a snack on hunger and food Kim YK, Kim JH, Ahn SM, Park JE intake Pharmaceutical & Health Research Institute, Pacific R&D King, N.A., Craig, S.A.S, Pepper, T., Blundell, J.E Center, Yongin-si, Korea University of Leeds, Leeds, UK

Aims: To clarify the anti-obesity effect of green tea and its Aims: To assess the independent and combined effects of active compounds. xylitol and polydextrose on appetite. Methods: Differentiated 3T3-L1 cells were treated with green Methods: 8 female and 7 male volunteers took part in four tea extract and its main active compounds (, conditions in a counterbalanced order. Each condition varied theanine and caffeine). Cellular viability, glycerol-3- according to the yogurt formulation administered as a snack phosphate dehydrogenase (GPDH) activity, glycerol release during the breakfast-lunch interval. The four conditions were: in the medium and HSL mRNA levels were checked. a control formulation (Cont) and three experimental Results: Glycerol release into the medium was significantly formulation yogurts: xylitol (Xyl), polydextrose (PDX) and increased by the cells treated with green tea extract. xylitol/polydextrose combined (Xyl/PDX). On d1 and d10 only, Theanine and caffeine from green tea also showed some the short and medium-term effects were assessed by level of lipolytic activity. Glycerol-3-phosphate measuring ad libitum lunch intake 90 minutes after the yogurt dehydrogenase activity was remarkably decreased by pre-load and by periodically measuring subjective ratings of catechins. In addition, green tea extract and its catechins hunger. caused an increase in the HSL mRNA levels. Results: The Xyl, PDX and Xyl/PDX yogurts induced a Conclusions: These results suggest that green tea has anti- significant suppression (10%) of food intake at the test lunch obesity effect through inhibition of adipogenesis by its compared with the control yoghurt. The PDX and Xyl/PDX catechins and stimulation of lipolysis by its theanine and yogurts also induced a significant suppression of hunger. This caffeine. Catechins, theanine and caffeine of green tea might suppression of appetite exerted by the xylitol and be the factors responsible for the modulation of lipid polydextrose formulations was due to the agents themselves metabolism. and not to any differential bias in energy content. Conclusions: Xylitol and polydextrose could be used as ingredients in functional foods for appetite control.

International Journal of Obesity Abstracts S54 T1:P1-045 T1:P1-046 Alterations of the functional features and carbohydrate Effect of insulin leptin and adrenalin on erythrocyte determinants (Cd) of erythrocytes (Er) membranes in Na++ /H antiport activity in obesity. women with abdominal obesity (AO). Zolota Z, Kaloyianni M, Paletas K and Koliakos G Kireev R.A., Kurmacheva N.A. Aristotle University, Thessaloniki Greece Saratov State University, Russia. Aims: The objective of our investigation was the studying of Aims: Na+/H+ antiport (NHE1) activity has been related to the Er acid resistance (functional changes) and the structural diabetes complications. Erythrocyte is an easily available cell changes of the membranes complexes carbohydrate that can be used for the study of insulin resistance in obesity. components in women with AO. Methods: NHE1 activity in erythrocytes derived from 10 Methods: We have studied the Er of 74 women (group 1) healthy individuals (BMI<25) and 10 obese patients with AO (the average age 29±0,98). The control group (BMI>30) has been estimated by intracellular pH consisted of 52 women (group 2) with simple obesity mesurements using 2'7'-bis(carboxyethyl)-5(6)-carboxy- (29±1,1). The acid resistance was studied by means of the fluoroscein tetraacetoxymethyl ester (BCECF) and by method of erythrograms. To determine Cd and structural estimating Na22 uptake in the presence of specific sodium changes of the membranes complexes, Er was incubated chanel inhibitors. Measurements in the presence of together with lectins. We have used the following (insulin, adrenalin or leptin) have been performed. preagglutination concentration of L: the β-galatose specific, 5-(N-Ethyl-Nisopropyl)-amiloride (EIPA) was used to inhibit PNA (125 mkg/ml); the mannose specific, ConA (125 NHE1 activity. Calphostin C (CC) inhibited Protein kinace C mkg/ml). Results: In group 1 there was discovered an and phorbol 12 myriststrate 12 acetate (PMA) was used to increase Cd of β-galatose, ready to react with PNA; with the stimulate protein kinase C activity. help of ConA there was observed an increase of the quantity Results: All three hormones and PMA enhanced NHE1 of the accessible receptors location of D- mannose. In group activity that was inhibited by EIPA and CC. Erythrocytes 2 there was not observed an interaction between lectins and derived from obese individuals showed an increased Cd of Er membranes. In the 1 group we noted a decrease of response to adrenalin but a blunted response to insulin and the Er membrane resistance than in group2. leptin. There was no difference in PMA response. Conclusions: The received results testify t o infringements in Conclusions: These differences may reflect changes of system of intercellular interactions, and will allow to erythrocyte hormone receptors in the obese state. understand mechanisms of formation o f an atherosclerosis.

T1:P1-047 T1:P1-048 Impaired Recruitment of Uncoupling Protein 3 Gene The change of plasma ghrelin in people with different Expression in Skeletal Muscle of Preterm Newborns breakfast meal hours Kopecky J, Brauner P, Kopecky P, Flachs P, Rovenska M Sangyeoup L, Yunjin Kim, Hyong-Hoi K, Han -Chul S, Tae- Institute of Physiology AS CR, Prague, Czech Republic Yong J, Mun-Sup S. Pusan National University, Busan, South Korea Aims: Experiments in mice suggest that postnatal recruitment of mitochondrial uncoupling protein 3 (UCP3) Aims: The purpose of this study is to evaluate the change of gene in skeletal muscle could be involved in maturation of plasma ghrelin levels in people with different breakfast hours. newborns and/or in “metabolic imprinting” by nutritional Methods: 4 healthy males (age range 23.4-35.5 years; BMI lipids. Our aim was to characterize the UCP3 expression in range 22.9-27.1kg /m 2) had fixed breakfast time or skipped muscles of human newborns. breakfast; however, they were asked to strictly keep their Methods: Transcript levels were measured using real-time breakfast time fixed and steady starting two weeks before RT-PCR in autopsy samples of skeletal and heart muscle of this study. Blood was drawn in EDTA tubes hourly from 0630 human, mostly preterm neonates, who died during the first until 1130. Ghrelin, leptin, insulin were measured by postnatal month (n = 28), and two aborted foetuses. radioimmunoassay. Nutrition and other clinical data were also recorded. Results: The plasma ghrelin level range of those who Results: Our results document postnatal induction of UCP3 consume their breakfast before and 2 hour after their gene expression in skeletal muscle, and involvement of breakfast was 113.0-800.0 pg/ml and 78.3-553.0 pg/ml. The nutritional lipids in the induction, and impairment of the leptin level range was 4.9-5.1 ng/ ml and 4.4-4.7 ng/ml . induction in neonates delivered before approximately 26 Compared to plasma ghrelin levels measured just before weeks of gestation. Very low expression of UCP3 was breakfast, it showed 7.2-30.9% decrease 2 hours after observed in the heart. breakfast. For subject who skips his breakfast, the plasma Conclusions: The results support a role of UCP3 in the ghrelin level was recorded the lowest at 0730. Conclusions: Plasma ghrelin levels were lowest 2 hours after postnatal activation of lipid oxidation in skeletal muscle. meals although breakfast meal hours were different. Plasma UCP3 may be involved in the delayed postnatal activation ghrelin level was lowest at 0730 in subject not consuming of oxidative metabolism in muscles of preterm neonates breakfast. and may represent a “metabolically imprinted” gene.

International Journal of Obesity Abstracts S55 T1:P1-049 T1:P1-050 Leptin and ghrelin levels in Cushing’s disease. Reduced expression of adiponectin mRNA in visceral Libè R, Morpurgo PS, Cappiello V, Barbetta L, Beck-Peccoz compared to subcutaneous adipose tissue. P and Spada A. Institute of Endocrine Sciences, University Lihn A S, Bruun JM, He G, Pedersen SB and Richelsen B. of Milan, Ospedale Maggiore, IRCCS. Milan (Italy). Aarhus Amtssygehus, Aarhus, Denmark.

Aims: To investigate leptin and ghrelin secretion in Cushing’s Aim: To investigate the gene expression and the production disease (CD). of adiponectin in subcutaneous compared to visceral Methods: evaluation of basal circulating levels of ghrelin, abdominal adipose tissue. insulin and leptin in 14 women (age±SD: 39.5±14 yrs, Methods: Paired adipose tissue biopsies from both normal- BMI±SD: 25.8±5.1 Kg/m²) with active CD. 14 healthy women weighted and obese women were investigated. Adiponectin matched for age and BMI (age 39.4±15, BMI: 26.8±5.5) were mRNA was determined by RT-PCR, and adiponectin used as controls. production by radioimmunoassay. Both fresh frozen adipose Results: Patients with CD showed lower levels of ghrelin tissue and cultured adipose tissue fragments were analysed. (117.8±80.6 vs 176±109.9 pmol/L, p<0.05), higher levels of Results: Fresh frozen adipose tissue adiponectin mRNA insulin (20.6±14 vs 12.7±8 U/L, p=NS) and leptin (27.6±8.5 levels were 33% lower in visceral compared to subcutaneous vs 18.5±19.5 ng/mL, p<0.05) compared to normal subjects. adipose tissue in normal-weighted subjects (p<0.05) and 29% Ghrelin levels were not correlated with urinary free cortisol, lower in visceral compared to subcutaneous adipose tissue in basal ACTH, cortisol, , , 17OH- obese subjects (NS). In normal-weighted and obese subjects , DHEA-S, leptin levels and BMI. visceral adipose tissue fragments cultured for 72 hours displayed 40-60% lower adiponectin mRNA levels compared Conclusions: The study confirms that leptin levels are high in to the level in subcutaneous adipose tissue fragments patients with CD, consistent with the view that (p<0.05). Adiponectin production in adipose tissue from glucocorticoids increase body weigh and load intake by obese subjects tended to be lowest in visceral adipose tissue limiting the central action of leptin. The central and/or fragments (0.32±0.14 vs. 0.52±0.11 µg/µg DNA, p=0.07). peripheral mechanisms responsible for the low levels of Conclusion: Adiponectin gene expression is lower in visceral ghrelin in the presence of glucocorticoid excess remain to be compared to subcutaneous adipose tissue, suggesting a elucidated. more important role for subcutaneous adipose tissue in regulating plasma adiponectin levels.

T1:P1-052 T1:P1-051 Adipose Tissue is a Source of Procalcitonin in Human Adiponectin Gene Expression in Subcutaneous Adipose Sepsis and in Inflammation Tissue of Obese Women: in Response to Short-term Very Linscheid P, Seboek D, Pierer G, Keller K, Müller B Low Calorie Diet and Refeeding Dept. of Research, University Hospitals, Basel, Switzerland Liu YM, Lacorte JM, Viguerie N, Poitou C, Pelloux V, Langin D, Basdevant A and Clément K. Aims: We explored potential -mRNA expression and Université Paris VI, Service de Nutrition Hôtel-Dieu, 75004 Paris procalcitonin synthesis in human adipose tissue. and INSERM U317, 31403 Toulouse, France Methods:Expression of CT-mRNA was analyzed by RT-PCR in adipose tissue, bowel and white blood cells from septic Aims: To investigate the effect of 48h Very Low Calorie Diet and control patients. Adipocyte cultures obtained from and subsequent refeeding on adiponectin (Ad) mRNA healthy subjects undergoing plastic surgery were treated with expression in sc adipose tissue of obese women. IFNγ, TNFβ, IL-1α and LPS. Procalcitonin release into culture Methods: Ad mRNA expression level in sc adipose tissue was supernatants was measured by sensitive chemiluminometric determined using a real time RT-PCR assay. Results: The VLCD caused a 33 % rise (P < 0.01) of average assay and calcitonin-mRNA was determined by quantitative ad mRNA level whereas refeeding caused a 32.8 % fall (P < real-time PCR. 0.05) of the mRNA average value. The change in Ad mRNA Results: Expression of CT-mRNA was demonstrated in level was negatively correlated to the change in weight loss human adipose tissue and in bowel obtained from septic (Rho = -0.62, P = 0.01) and positively correlated to free fatty patients with increased circulating procalcitonin, but not in acid plasma level change (Rho = 0.79 P = 0.02) after VLCD. white blood cells. In vitro, expression of calcitonin-mRNA The subjects who showed acute Ad mRNA response had was increased 315-fold (p<0.01) by IL-1β, and 200-fold by higher levels of Ad mRNA before and after VLCD (P = 0.02 and LPS (p<0.01) after 6 hours. Procalcitonin protein secretion P = 0.003, respectively) and a borderline significantly lower BMI into culture supernatant was increased 8.7-fold (p<0.05) by (P = 0.1) compared with the subjects who showed no Ad mRNA IL-1β, and 6.8-fold (p<0.05) by LPS after 48 hours. response. A significant increase (P = 0.008) of insulin sensitivity Conclusions: The data presented herein strongly suggest an of the responder subgroup was found after VLCD. active role of adipose tissue in inflammation and sepsis- Conclusion: This study indicates that Ad can acutely respond to related increase in plasma procalcitonin. short-term energy changes in some obese subjects. The level of adiposity and insulin sensitivity may contribute to the modulation of this response.

International Journal of Obesity Abstracts S56 T1:P1-053 T1:P1-054 Energy Expenditure In Obese Men And Women Measured Chronic effects of Cannabinoid 1 (CB1) Receptor Using [14C]-Bicarbonate-Urea: Effect Of Weight Loss. Inverse Agonist, AM251, on indices of energy balance N.Luscombe, I.Kirkwood, M.Bellon, C.Tsopelas, G.A.Wittert. and gastrointestinal motility in mice Dept. of Medicine, University of Adelaide, Australia, 5000. Shearman, L, Rosko, K, Camacho, R, Stribling, S, Zhou, D, Kuca, S, Strack, A, MacIntyre, DE. Aims: The process of weight loss or the maintenance of Merck Research Laboratories, Rahway, NJ, USA reduced weight has been shown to either reduce or have no effect on total energy expenditure [TEE]. The aim of this study Aims: To evaluate the effects of chronic administration of the was to determine the effect of an 8-week weight loss program CB1 inverse agonist, AM251, on food intake, body weight, followed by 2-weeks of weight maintenance on TEE using body composition and gastrointestinal (GI) motility in mice. [14C]-bicarbonate-urea. Methods: Dietary-induced obese mice were given AM251 (1 Methods: 6 men and 5 women, mean (±SE) age 50±3 yrs, BMI or 10 mg/kg/day, po) for 30 days. Food intake and body 2 34.1±2.1 kg/m and body fat 38.7±3% were studied before and weight were measured daily; body composition (by NMR) 2 weeks (while weight stable) after an 8% or greater weight was measured pre-study and at termination. GI motility was loss. Weight loss was induced using an ~3.34 MJ/day, 50% measured by charcoal transit in lean mice after single or protein, 37% carbohydrate, 13% fat diet. Body composition sequential (7) daily doses of AM251 (10 mg/kg, po). [DEXA], resting energy expenditure [REE] and the thermic Results: AM251 elicited dose-dependent decreases in daily effect of a 2.7 MJ test meal (18% protein, 62% carbohydrate, food intake, cumulative food intake, body weight and fat 20% fat) [TEF] (indirect calorimetry) were measured in mass. The evoked decreases in body weight plateaued at addition to TEE. Nutrient intake and physical activity were days 5 – 12, and were sustained. Food intake effects were monitored using diaries. maximal at days 1 - 4 and later declined. Enhanced GI Results: Body weight decreased by 11.8 ± 1.0%. Total fat motility was evident following initial exposure to AM251 but mass, abdominal fat mass and lean mass decreased by 19.7 ± not following chronic exposure. 1.8%, 21.8 ± 2.3% and 6.0 ± 1.1%, respectively. At week 12, Conclusions: Although weight loss evoked by chronic AM251 REE decreased by 5.7 ± 1.3% (P < 0.002). Changes in TEF (- treatment is sustained, food intake inhibition and gastric 13.3 ± 13.7%) and TEE (-0.18 ± 3.68 %) were not significant. prokinetic effects undergo tachyphylaxis. Anti-obesity effects Conclusion: In free-living subjects, after weight loss and two of AM251 involve mechanisms other than anorexigenesis, weeks of weight maintenance, there is a decrease REE but such as increased energy expenditure. not TEF or TEE.

T1:P1-055 T1:P1-056 Effects of a β3-adrenergic agonist on adiponectin Differentially expressed genes in cafeteria-fed rats expression in lean and overweight (cafeteria) rats. López IP, Marti A, Milagro FI, Moreno MJ, Martínez JA, De Moreno-Aliaga MJ, Urresti J, Lamas O, Marti A, Martínez JA Miguel C University of Navarra, Pamplona, Spain Universidad de Navarra, Pamplona. Spain.

Aims: Adiponectin is a hormone produced by adipocytes, Aims: Identification of WAT genes differentially expressed in which has been related to insulin sensitivity. The aim of this an obesity model. study was to find out if the potential anti-diabetic properties Methods: Epididymal WAT was obtained from rats fed on of the 3-adrenoceptor agonist Trecadrine might be related control and cafeteria diets for 65 days. RNA from three rats to changes in adiponectin gene expression levels. of each group was pooled and hybridized to Rat Genome- Methods: Control and cafeteria-fed rats were treated with U34A Array (Affymetrix) containing 4500 genes and 8000 placebo or Trecadrine during 35 days. RNA was extracted EST clusters. The data were confirmed by Northern blot with from retroperitoneal adipose tissue using Trizol. Adiponectin leptin, UCP3 and PPARγ in three additional animals of each mRNA was determined by Northern blot and normalized with dietary group. Results: Altered gene expression patterns respect to the 18S ribosomal signal. affecting 15% of the sequences were seen. Thus; 416 genes Results: Trecadrine treatment prevented the increase in were induced more than 2-fold and 429 were repressed body and fat weights observed in rats fed on a cafeteria diet. more than 2-fold in WAT from cafeteria-diet fed animals. The obese cafeteria-fed rats showed a mild Genes among them are involved in functions such as: , which was prevented by Trecadrine. macronutrient metabolism, transcription regulation, Serum glucose levels were similar in all groups. Adiponectin hormones and signal transduction, cytoskeleton and redox or mRNA was significantly decreased in cafeteria overweight stress related proteins. In general, genes implicated in lipid rats. Trecadrine treatment did not have any significant effect metabolism were up-regulated while redox and stress related on adiponectin gene expression. genes were down-regulated in WAT of obese animals. Conclusions: The development of hyperinsulinemia and Conclusions: The cafeteria model of diet-induced obesity is insulin resistance induced by high fat intake could be useful for the characterization of the molecular mechanisms secondary, at least in part, to the fall in adiponectin levels of this disorder and can help to understand human obesity observed in these rats. The known antidiabetic properties of cases that are thought to occur as a consequence of a high Trecadrine are not mediated by changes in adiponectin. fat intake.

International Journal of Obesity Abstracts S57 T1:P1-057 T1:P1-058 Effect of abdominal fat deposits on the response to Involvement of AMP-activated protein kinase in atorvastatin treatment in familial hypercholesterolemia. obesity resistance induced by respiratory uncoupling Martínez-Valls J, Civera M, Sanz J, Real JT, Chaves FJ, in white fat Priego A, Ascaso JF, Carmena R. Servicio de Matejkova O, Flachs P, Sponarova J, Rossmeisl M, Miksik I, Endocrinología y Nutrición Hospital Clínico, Universidad de Thomason-Hughes M, Hardie D G and Kopecky J Valencia. Institute of Physiology AS CR, Prague, Czech Republic

Aims: To study the influence of the type of obesity on Aims: Transgenic (aP2-Ucp1) mice with ectopic UCP1 in lipoprotein phenotype and response to 20 mg day of white fat are resistant to obesity. The transgene reduces atorvastatin in women with molecularly defined familial ATP/ADP ratio, elevates oxygen consumption, decreases hypercholesterolemia (FH). fatty acid synthesis, mitigates lipolysis and induces Methods: We have studied 34 FH women (mean age 43.2 mitochondrial biogenesis in adipocytes. A hypothesis was years, BMI 25.5 ± 4.58 and WHR 0.79± 0.16). In all subjects tested whether the complex effect of the UCP1 is mediated in basal and after 8 weeks of atorvastatin treatment by activation of the AMP –activated protein kinase (AMPK). anthropometric parameters and lipids were measured with Methods: ATP and AMP contents (HPLC) and AMPK activity standard methods. The genetic diagnosis of FH was (specific peptide phosphorylation assay) were estimated in established with PCR-SSCP and Southern blot analysis. adipose tissue of control and transgenic mice. The Results: The abdominal fat deposit (WHR > 50 th of the expression of PPAR-γ and aP2 were also analyzed. whole group), negatively influence blood pressure (SBP Results: ATP/AMP ratio in adipocytes from subcutaneous fat 114.1±13.6 vs 131.47± 16.8 y DBP 70.8±10.6 vs 82.3±7.31) of the transgenic animals was significantly lower than in and basal plasma TG levels (77.5±37.2 vs 124.6±48.4). In controls. The presence of transgenic UCP1 resulted in addition, the response of LDLc values after atorvastatin approximately a 2-fold increase of AMPK activity in treatment, was significantly lower in FH women with a WHR subcutaneous fat. A significant diminution of PPAR-γ and > 0.79 (42.9 ± 8.3% vs 32.9 ± 13.5%). aP2 mRNA levels was found in transgenic animals. Conclusions: The abdominal fat deposit has a negatively Conclusions: AMPK is involved in the obesity resistance of influence on blood pressure, plasma TG levels and the aP2-Ucp1 mice and may have important role in adipose decrease of LDLc values after atorvastatin treatment. tissue biology.

T1:P1-059 T1:P1-060 CHANGES IN MUSCLE MYOSTATIN EXPRESSION IN Ghrelin – a new potential factor reducing blood OBESE SUBJECTS AFTER WEIGHT LOSS . pressure in weight loss treatment Milan G., Granzotto M., Mingrone G., Manco M., Greco A.V., Mizia-Stec K, Zahorska-Markiewicz B, Olszanecka- Federspil G., Vettor R. Glinianowicz M, Janowska J, Mucha Z; Dept. of Medical and Surgical Sciences, Padua, Italy. Silesian University School of Medicine, Poland

Aims: Myostatin belongs to TGF-β superfamily and acts as a Aims: In animal models ghrelin reduces cardiac afterload muscle growth inhibitor. Myostatin-null mice show an and increases cardiac output via receptors in cardiovascular increase in skeletal muscle mass and a reduction in fat system The aim of our study was to evaluate a potential accumulation with age. Myostatin administration in mice relationship between weight loss treatment, blood pressure results in a remarkable skeletal muscle and white adipose and serum ghrelin concentrations in obese women. tissue loss. We studied skeletal muscle myostatin mRNA Methods: 27 obese patients (BMI: 36.5+5 kg/m2) and 11 levels in obese patients before and after weigth loss due to controls (BMI: 22.2+2 kg/m2) were involved in the study. biliopancreatic diversion (BPD). Blood pressure and serum ghrelin levels were assessed Methods: Changes in body composition, as assessed by 3H- before and after a 3 month-weight reduction treatment. water, insulin sensitivity, measured by clamp, and urinary Results: There were not differences between plasma nitrogen were investigated in 5 obese subjects before and ghrelin concentrations in obese patients and controls. after BPD. Myostatin mRNA was quantified by real-time RT- Following weight loss (mean 8.7±4.5kg) SBP decreased PCR in muscle biopsies. (120+13 vs 115+14 mmHg, p=0.01) and serum ghrelin levels Results: After BPD treatment the patients had lost 37% of increased significantly (66.9±13.7 vs 73.9±15.4pg/ml; their initial body weight, reducing their FM and also their p=0.005). There were significant correlations between: FFM. Daily urinary nitrogen excretion was slightly increased post-weight loss ghrelin levels and SBP (r= -0.450, p=0.018), after BPD. Myostatin mRNA levels resulted clearly DBP (r= -0.415, p=0.031); ghrelin levels and ∆SBP (r= decreased in muscle tissue after BPD. 0.516, p=0.006), ∆DBP (r=0.529, p=0.005) Conclusions: Our results suggest a role for myostatin in the Conclusions: It is the first evidence in humans that weight regulation of muscle mass changes in humans. We cannot loss might decrease blood pressure in obese patients by exclude that the down-regulation of Myostatin may be linked ghrelin dependent mechanism. to FM loss and insulin sensitivity improvement we observed.

International Journal of Obesity Abstracts S58 T1:P1-061 T1:P1-062 Tolerance does not develop after chronic infusion of Rapid desensitization of lipolysis in rat white adipocytes mCPP on food intake and body weight gain in rats pretreated with β-adrenergic agonists Ashkzari M, Modiri A-R and Svartengren J Mori S, Nojiri H, Yoshizuka N, Takema Y, *Saito M Biovitrum AB, Department of Biology, Stockholm, Sweden Biological Science Laboratories, Kao Corporation, Tochigi 321-3497, Japan

Aims: The role of the serotonin 5-HT2C receptor in feeding behaviour has been established in numerous studies. This Aims: In adipocytes, transient and prolonged stimulation of - investigation was undertaken to study tolerance development adrenergic receptor (βAR) induce desensitization to on food intake and body weight in rats following chronic catecholamines leading to a decrease in intracellular treatment with the non-selective 5-HT2C receptor agonist, accumulation of cAMP. We assessed catecholamine-induced mCPP. desensitization of three βAR subtypes by examining lipolytic Methods: Male Sprague-Dawley rats were implanted with response of rat white adipocytes. osmotic mini-pumps and mCPP (3, 10 and 30 mg kg-1 day-1) Methods: Adipocytes isolated from epididymal fat pad of or saline were infused for 14 days. Food intake and body Wistar rats were pretreated with isoproterenol (ISO). After weight were registered once a day. Treatment was washing the cells, their lipolytic response to various withdrawn on day 14 but measurements continued. Acute concentrations of NE, dobutamine (β1 agonist), terbutaline challenge of mCPP (30 mg kg-1 s.c.) in drug naive and drug (β2 agonist), or BRL37344 (β3 agonist) were examined by treated rats was done before and after infusion. measuring released glycerol into the incubation medium. Results: Chronic infusion of mCPP produced a dose- Results: When adipocytes were pretreated with ISO, NE- dependent and sustained reduction in body weight gain. induced lipolysis was significantly reduced in dose (10 nM- Food intake decreased also but attenuated over time 100 µM)- and time (5 – 30 min)-dependent manners. No indicative of tolerance development. However, acute noticeable difference was found in the lipolytic response to challenge with mCPP before and after infusion produced dobutamine, terbutaline, and BRL37344 in pretreated cells. similar reduction in food intake. Conclusions: Transient stimulation of AR by non-selective Conclusions: Tolerance appears not to develop even at high -agonists reduced lipolytic response to selective 1, 2 and mCPP doses, thus it is not likely the cause to the attenuated 3 agonists in rat white adipocytes, suggesting some food intake. Body weight reduction persists even after 14 common mechanism for agonist-induced rapid days withdrawal. desensitization of three AR subtypes.

T1:P1-063 T1:P1-064 Ghrelin secretion after standard meal test in obese Effects of tea catechins on diet-induced obesity in mice subjects before and after weigh loss Murase T, Nagasawa A, Suzuki J, Hase T, Tokimitsu I Morpurgo PS, Resnik M, Agosti F, Cappiello V, Sartorio A, Biological Science Laboratories, Kao Corporation, Japan Beck-Peccoz P, Spada A. Institute of Endocrine Sciences, University of Milan, Ospedale Maggiore, IRCCS. Milan (Italy). Aims: We investigated the effects of long-term feeding with tea catechins, which are naturally occurring polyphenolic Aims: To investigate response of ghrelin to a standard meal compounds widely consumed in Asian countries, on the test in obese subjects before and after weight loss. development of obesity in C57BL/6J mice. Methods: evaluation of circulating levels of ghrelin, insulin Methods: We measured body weight, adipose tissue mass, and leptin in response to a meal test at baseline and after 3 and liver fat content in mice fed diets containing either low-fat weeks body weight reducing program (BWR), in 10 obese (5% triglyceride (TG)), high-fat (30% TG), or high-fat subjects (3 male, 7 female; mean age 35±9.3 yrs, BMI supplemented with 0.1-0.5% (w/w) tea catechins for 11 45.2±10.6 kg/m2). 5 normal weight subjects were used as months. The β -oxidation activities and related mRNA levels controls. were measured after 1 month of feeding. Results: At baseline, obese subjects showed significantly Results: Supplementation with tea catechins resulted in a lower ghrelin levels than controls, which were negatively significant reduction of high-fat diet-induced body weight correlated with BMI, weight, insulin and leptin levels. Fasting gain, visceral and liver fat accumulation, and the ghrelin levels were not modified by standard meal test in development of hyperinsulinemia and hyperleptinemia. obese (from 110.8±69.7 to 91.8±70.2 nmol/L, p=NS), while a Feeding with tea catechins for 1 month significantly significant reduction was observed in controls. After the BWR increased ACO and MCAD mRNA expression as well as β- program obese subjects significantly reduced weight, BMI, oxidation activity in the liver. and leptin levels, while no significant changes were found in Conclusions: The stimulation of hepatic lipid metabolism both fasting ghrelin and ghrelin response after the meal. might be a factor responsible for the anti-obesity effects of Conclusions: 5% weight loss obtained after a short-term tea catechins. The present results suggest that long-term period of integrated BWR program is not sufficient to consumption of tea catechins is beneficial for the normalize fasting ghrelin levels nor to restore the normal suppression of diet-induced obesity, and it may reduce the ghrelin suppression after a meal in severely obese subjects. risk of associated diseases including diabetes and CHD.

International Journal of Obesity Abstracts S59 T1:P1-065 T1:P1-066 Ab Effects of various diets on rat mesenteric in vitro. Effect of weight loss on serum concentrations of nitric Naderali EK, Sadler C, Williams G. University of Liverpool oxide, TNF - and soluble TNF- receptors Olszanecka-Glinianowicz M, Zahorska-Markiewicz B, Aims: Diet is thought to play a role in vascular reactivity. In this Janowska J., Zurakowski A, study we have examined the effects of 3 different diets on metabol- Department of Pathophysiology, Silesian University School of ic changes and vascular function in male Wistar rats. Medicine, 40 – 752 Katowice, Poland Methods: Rats were fed either standard laboratory chow through- out(n=5; control) or given chow plus chocolatebiscuit (n=5; biscuit- Aims: Determine whether weightloss modulates NO, TNF-α fed) or a 45% fatdiet (n=5; high-fat) for 10 weeks. and sTNFR1and sTNFR2 serum concentrations. Results: Biscuit- and high-fat-fed animals had significantly higher Methods: 43 obese women ( age 41,8 ± 11,9 y, BMI before body weights (30%, p<0.01) and white adipose tissue mass (195- 36,5 ± 4,6 kg/m2 after 33,4 ± 4,6 kg/m2 ) were examined 274%, p<0.01),but not gastrocnemius and brown adipose tissue before and after 3 months weight reducing treatment, mass, than the control rats. Biscuit-fed, but not high-fat-fed rats consisting of 1000 kcal/day diet and physical exercises. had significantly raised plasma levels of free fatty acids (150%, α p<0.01) and triglyceride (240%, p<0.01) than chow-fed group. Serum concentrations of NO, TNF - and sTNFR1 and sTNFR2 Plasma levels of glucose,insulin and leptin were similar in all three were measured by ELISA. groups (p>0.5).Vasoconstrictions to noradrenaline and KCl were Results: ****p< 0,00001; *****p<0,0000001 significantly higher in mesenteric arteries from biscuit-fed but not NO TNF - STNFR1 STNFR2 high-fat-fed rats, than the controls (for both P<0.05) while vasore- µmol/l pg/ml pg/ml pg/ml laxations to acetylcholine and sodium nitroprusside, but not hista- before 33,5± 10,1 6,9 ± 2,4 1237,2± 226 1782,7 ± 417 mine, were markedly reduced and EC50 values shifted to the right **** ***** ***** in both biscuit- and high-fat-fedgroups (for all, p<0.05). after 35,6± 13,5 5,4 ± 1,6 1432,8± 305 2063,0 ± 341 Conclusions: High-fat and high-energy diets induce obesity that re- sults in metabolic and vascular dysfunction. Diet-induced vascular In multiple regression analysis significant association NO ∆ ∆ α ∆ ∆ dysfunction appears to be selective and affects cGMP-mediated with BMI, TNF- with mass adipose tissue (kg), TNFR1 ∆ NO generation pathway, while cAMP-mediated NO production with body mass were found. pathway appears to be unaffected. Conclusions: Decrease of TNF-α and increase in sTNFR1, sTNFR2 and NO serum concentrations after weight reducing treatment, may be a contrregulation preventing further weight loss.

T1:P1-067 T1:P1-068 Influence of 7 days overfeeding with a low or normal EXPRESSION OF GENES OF LIPID METABOLISM IN protein diet on weight gain and energy expenditure HEART OF GENETICALLY OBESE ZUCKER RATS. J.M. Oomen, M.A. van Baak, and W.H.M. Saris Pagano C, Calcagno A, Granzotto M, Federspil G, Vettor R. Department of Human Biology/NUTRIM, Maastricht Via Ospedale 105, University of Padova, Italy University, The Netherlands Aims: Heart of Zucker diabetic fatty (ZDF) rats display a Aims: The purpose of this study was to examine weight gain preferential shift of excess fatty acids to triglycerides (TG) and changes in energy expenditure (EE) during short-term that accumulate to produce cardiac lipotoxicity. The aim of overfeeding with diets differing in protein content in humans. these study was to investigate if similar abnormalities of lipid Methods: Ten normal weight male subjects were overfed two metabolism are present in genetically obese rats without times 7 days with 1000 kcal extra per day (Normal Protein diabetes. Methods: 8 fa/fa Zucker rats (obese), 10 lean FA/? Diet (NPD): 15En% protein, 35En% fat and 50En% Zucker rats (lean) and 6 diet-restricted fa/fa rats with 20% carbohydrates (CHO); Low Protein Diet (LPD): 3En% protein, reduction of body weight (post-obese) were studied. Heart 30En% fat and 67En% CHO). Diets were given in random was collected after decapitation and immediately frozen in order with 3 weeks in between. Body weight, body liquid nitrogen for determination of gene expression by RT- α composition and EE (respiration chamber: total EE; PCR. Results: Genes associated to lipid oxidation (PPAR- , ventilated hood: BMR) were measured at the start and the ACO, CPT-1, UCP2 and UCP3) were all significantly end of the overfeeding period. increased in obese compared to lean rats, while expression Results: Weight change differed between the two diets (NPD of GPAT, a key step in TG synthesis, was unchanged. In α 0.65 ± 0.20 kg and LPD -0.17 ± 0.21 kg, p < 0.01). Total EE post-obese rats PPAR- was reduced to levels comparable did not change significantly with overfeeding and no to lean rats while ACO, CPT1 and UCP3 remained differences in change between the diets was found. BMR significantly increased; GPAT was increased compared to increased slightly, but significantly, during LPD, but not both obese and lean rats. Conclusions: In contrast to data during NPD. reported in diabetic rats, our data suggest that, in fa/fa rats, Conclusions: Overfeeding a NPD resulted in weight gain, increased fatty acids availability is associated to enhanced LPD did not. These differences could not be explained by myocardium lipid oxidation rather than increased TG differences in EE, as measured in the respiration chamber. deposition. Our data suggest a preferential channelling to TG synthesis in of post-obese rats.

International Journal of Obesity Abstracts S60 T1:P1-069 T1:P1-070 Adiponectin Levels in Patiens With Short term effect of low- and high-carbohydrate diets on Parzková J, Kržová J, Kotrlíková E, Lacinová Z , Justová V, plasma leptin levels upon cold exposure in humans Papežová H and Haluzík M. 3rd Department of Medicine, 1 Parker T, Imbeault P, Doucet E, Weber JM and Haman F Faculty of Medicine, Prague, Czech Republic. School of Human Kinetics, University of Ottawa, Ottawa, Canada. Aims: Adiponectin (ADP) is an adipocyte-derived plasma protein that increases insulin sensitivity in the muscle and Aims: To examine the effect of glycogen availability on liver and may represent a link between obesity, insulin plasma leptin levels before and after cold exposure in resistance and atherosclerosis. The aim of our study was to humans. compare serum ADP levels in patients with anorexia nervosa Methods: 6 healthy men (age: 22 ± 1 y; % body fat: 13 ± 1) (AN) and age-matched healthy females (C). were exposed to 100C (liquid conditioned suit) for 90 min Methods: Serum concentrations of ADP, leptin, soluble leptin after two different 3 day isocaloric diets: 1) low carbohydrate receptor (S-LEPR), TNF-α and cortisol were measured in 15 (LCHO = 68 g/d) with daily exercise bouts (1 h at 65% of VO2 females with AN and 18 age-matched healthy C subjects. max) and; 2) high CHO (HCHO = 494 g/d) without exercise. Results: BMI was significantly reduced in AN patients relative Serum leptin was assayed after an overnight fast and to C. Serum ADP levels in AN were significantly increased following 90 min of cold exposure after each diet. relative to C (48,26±17,14 vs 33,9± 13,36 µg/ml). Serum Results: After the LCHO diet, leptin levels decreased leptin concentrations were reduced in AN, while serum significantly in response to cold exposure (0.53 ± 0.21 to soluble levels were significantly higher than in 0.34 ± 0.25 ng/ml, P < 0.05). Leptin levels also decreased in C. TNF-α and cortisol levels did not differ between groups. response to cold following the HCHO diet, but this did not Conclusions: Plasma adiponectin and S-LEPR levels were reach statistical significance (0.71 ± 0.40 to 0.55 ± 0.42 increased in patients with AN relative to C, while BMI and ng/ml, NS). serum leptin were markedly lower. Hyperadiponectinemia in Conclusions: These results support the notion that the AN patients is most likely the result of reduced body fat activation of the sympathetic nervous system during cold content. The physiological consequences of this finding need exposure decreases circulating leptin levels in humans. This to be clarified. (Supported by grant GACR No. 6663-3) occurs when glycogen stores are a priori depleted through a LCHO diet combined with exercise bouts.

T1:P1-071 T1:P1-072 Anti-glucocorticoid effects of progesterone on rat Resistin expression in different adipose tissue depots adipose tissue metabolism in vivo. during rat development Pedersen SB, Kristensen K, and Richelsen B. Oliver P, Picó C, Serra F, Palou A Aarhus Amtssygehus, Aarhus, Denmark. Universitat de les Illes Balears, Palma de Mallorca, Spain

Aim: To investigate the possible anti-glucocorticoid effects of Aims: To establish the ontogenic pattern of resistin progesterone on rat adipose tissue metabolism in vivo. expression in different white adipose tissue depots (WAT) - Methods: Male rats were adrenectomized divided into four epididymal, inguinal, mesenteric and retroperitoneal - and in groups 1. saline (control), 2. (dexa), 3. (BAT), in rats of different ages (from progesterone (prog), 4. dexa and prog for three weeks. the suckling period to one year of age). Adipose tissue LPL-activity and lipolysis were analysed. Methods: Resistin mRNA was determined by Northern Results: Dexa-treated rats had lower body weight, lower Blotting, and serum resistin levels by enzyme immunoassay. amount of adipose tissue, and higher blood-glucose, than Results: Resistin expression remains almost constant with control rats. Concomitant prog-treatment, had no influence on age in WAT, except in early development, where there is a body weight, blood-glucose, but prog attenuated the decrease peak of expression in the epididymal and retroperitoneal in amount of adipose tissue. Furthermore, LPL-activity was depots, and a decrease in the inguinal one. Moreover, there increased in adipose tissue from dexa-treated rats. This dexa- is a site-specific difference regarding resistin expression: the induced stimulation could be blocked by prog-treatment. depots express characteristic levels of mRNA, especially at Finally, dexa-treatment was associated with an increase in the age of 2 months, moment when resistin mRNA levels are lipolysis, which also was blocked by prog-treatment of the significantly higher in the epididymal and retroperitoneal than rats. in the inguinal and mesenteric WAT and than in the BAT. Conclusion: In male rat adipocytes (which does not contain Circulating resistin levels increase with age probably progesterone receptors), progesterone acted as an anti- reflecting the increase in the body fat content. glucocorticoid and inhibited the dexa-induced increase in Conclusions: The transient increased resistin expression in lipolysis and LPL-activity. It is suggested that progesterone the epididymal and the retroperitoneal WAT at a period of also in humans might inhibit glucocorticoid effects on human time in which there is a change in diet suggests a nutritional adipose tissue metabolism, and thus in women might protect regulation of the resistin gene in both depots. against central accumulation of adipose tissue.

International Journal of Obesity Abstracts S61 T1:P1-073 T1:P1-074 Ghrelin variations during cold exposure after low- FLUOXETINE ALTERS ALPHA-MELANOCYTE and high- carbohydrate diets in lean men. STIMULATING HORMONE IN THE OBESE ZUCKER Pomerleau M, Doucet E, Weber JM, and Haman F. RAT University of Ottawa, Ottawa, Canada Churruca I, De la Cruz V, Morón L, Echevarría E, Macarulla MT, Abecia LC, Portillo MP Aims: To determine the effects of cold exposure on ghrelin University of the Basque Country, Vitoria, Spain levels after low- and high- carbohydrate diets (LCHO AND HCHO). Aims: The purpose of the present work was to describe the Methods: 6 healthy men (age: 22 ± 1y; % body fat: 13 ± 1) regional effects of fluoxetine on α-MSH immunostaining in were exposed to 100C (liquid conditioned suit) for 90 min the obese Zucker rat hypothalamus. after two consecutive 3-day isocaloric diets starting with: 1) Methods: fa/fa Zucker rats were administered with i.p. LCHO (~70 g/d) with 1 h/day exercise sessions at 65 % of fluoxetine chlorhydrate (10 mg/kg/d), for two weeks. The VO2max and; 2) a HCHO (~500 g/d) without exercise. Serum control group was given 0.9% NaCl. 50 µm brain sections ghrelin was assayed after an overnight fast and after 90 min were immunostained for α-MSH by the avidin-peroxidase of cold exposure at the end of each diet. technique and both the numbers of positively stained neural Results: After the overnight fast, ghrelin levels tended to be cells in arcuate nucleus and optical density in other higher following the LCHO than the HCHO (3508 ± 900 vs hypothalamic regions showing immunostained neural fibers, 2158 ± 280 ng/ml, respectively; ns). Ghrelin levels increased were measured with the aid of a computerized image after 90 min of cold exposure following the HCHO (2158 ± analysis system. 147 vs 3217 ± 397 ng/ml, respectively) while no difference in Results: Fluoxetine administration significantly decreased ghrelin levels was observed after 90 min of cold exposure food intake (26%) and body weight (54%). A significant following the LCHO diet. increase in α-MSH immunostaining was observed in the Conclusions: To our knowledge, this is the first study to arcuate nucleus and lateral hypothalamic area, without demonstrate the effects of cold exposure on ghrelin levels in changes in other hypothalamic regions. human subjects. Whether these changes are accompanied Conclusions: These results suggest the involvement of α- by concomitant changes in appetite under these conditions MSH in the anorectic mechanism of this drug in obese will need to be further investigated. Zucker rats. Supported by Basque Government (GV PI-1999-51)

T1:P1-075 T1:P1-076 Mechanism of Action of Azaftig (AZ): A Novel Depletor Sympathetic stimulation by cold exposure and fasting of Fat Depot J Figueroa1, P Vijayagopal1, T Hosoya1, G Mitchell2, and Rayner DV, Duncan JS and Do M-S C.Prasad1 Rowett Research Institute, Aberdeen, Scotland, UK 1Dept of Medicine, LSUHSC, New Orleans, LA, USA; and 2Dept of Pediatrics, Sainte-Justin Hospital, Montreal, Quebec, Aims: Fasting and cold exposure have been used to Canada. sympathetically stimulate adipose tissue The aim of this work is to examine the relative effects of fasting and cold exposure Aims: To examine the mechanism through which AZ, a on circulating catecholamine levels urinary proteoglycan derived from cachectic patients, causes Methods: Mice (Aston strain) were cold-exposed, fasted or increase in lipolysis and loss of fat depot. cold exposed and fasted. Noradrenaline (NA) and Methods: Lipolysis in adipocytes from 3T3-L1 cells and (A) were measured in plasma by radioimmunoassay (Tricat, mouse fat were measured by glycerol release. cAMP levels IDS Newcastle) and UCP1 gene expression in BAT by were determined by RIA. Northern blotting. Results: A pathway involving cAMP and hormone-sensitive Results: Fasting (24h) increased NA from 0.28+0.02 to lipase (HSL) is the major regulator of lipolysis in adipocytes. 0.44+0.06 ng/ml and A from 0.12+0.05 to 0.18+0.04 ng/ml. To examine whether AZ acts through this pathway, we Cold exposure (8 or 24h) increased NA to 0.74+0.11 or measured the effect of AZ on cAMP production by 3T3-L1- 0.71+0.10 ng/ml and A to 0.18+0.07 or 0.30+0.09 ng/ml). adipocytes. While treatment with IBMX, a phosphodiesterse Fasting (24h) plus cold (8h) gave similar results to fasting inhibitor, raised cAMP and stimulated lipolysis, AZ caused a alone (NA 0.48+0.08, A 0.19+0.09 ng/ml). UCP1 mRNA in decrease in cAMP in the face of increased lipolysis. BAT decreased on fasting, increased oncold exposure, but Therefore, AZ may stimulate lipolysis through a mechanism was unchanged during fasting with cold exposure. independent of HSL. This is supported by diminutions in Conclusions: Sympathetic responses vary with the stimulus: lipolysis stimulation by beta-adrenergic agonist but not AZ in the adrenal medulla contributes to the response to cold HSL -/- compared to HSL +/+ adipocytes. exposure, but has a lesser role in fasting or in fasting plus Conclusions: AZ promotes lipolysis through a novel cold exposure. In the fasting response, the overflow of NA mechanism independent of HSL. from adrenergic nerves appears to predominate.

International Journal of Obesity Abstracts S62 T1:P1-077 T1:P1-078 Variation of plasma resistin in obesity Opposite actions of testosterone and progesterone on UCP1 mRNA expression in cultured brown adipocytes Rayner DV, Kim, K-W, Do M-S, Hunter L, Duncan JS and Rodríguez AM, Monjo M, Roca P, Palou A Hoggard N University of the Balearic Islands, Palma de Mallorca, Spain Rowett Research Institute, Aberdeen, Scotland, UK Aims: To investigate the effects of the main physiological sex Aims: Resistin secreted by adipose tissue was recently hormones on rodent brown adipocytes. reported as decreasing insulin sensitivity in the liver. This Methods: Brown adipocytes were grown in primary culture work examines resistin levels in lean, obese and diabetic and treated with sex hormones and their inhibitors. obese Koreans and in Zucker rats (10 and 26 weeks old). Results: Testosterone-treated cells showed fewer and Methods: Resistin was measured by radioimmunassay smaller lipid droplets than controls and a dose-dependent (Phoenix Pharmaceuticals - antibody to resistin (43-65)-NH2) inhibition of UCP1 mRNA, under noradrenaline (NA) and resistin mRNA in rat adipose tissue by Northern blotting. stimulation. These effects were reverted by the androgen Results: Serum resistin was higher (P=0.02, n=7) in diabetic flutamide, suggesting the involvement of obese Koreans (1.70+0.03 ng/ml, BMI 26.2+0.6 kg.m-2), this receptor. Progesterone- and 17- --treated cells compared to the lean and simple obese (1.51+0.06, had more and larger lipid droplets. Progesterone stimulated 1.62+0.09 ng/ml, BMI 20+0.7, 32.7+1.5 kg.m-2). Plasma NA-induced UCP1 mRNA at the lower concentration tested, resistin was less (1.14+0.05 ng/ml, P=0.01) in young obese but not at higher concentrations, showing a dose dependent rats (BW 348g) compared to young lean, old lean and old effect. Interestingly, the progesterone receptor antagonist obese rats (BW 269, 508, 672g, resistin 1.38+0.07, RU486 induced UCP1 and UCP2 mRNAs, suggesting it has 1.38+0.02, 1.37+0.03 ng/ml). Resistin mRNA in epididymal a key effect on brown adipocyte thermogenic capacity. white adipose tissue was less (P<0.01) in all obese rats. Conclusions: Testosterone, 17- -estradiol, progesterone and Conclusions: Increased serum resistin may be associated RU486 have distinct actions on brown adipocytes, thus with insulin resistance in Koreans. In contrast, the decrease modulating UCP1 and UCP2 mRNA expression and/or lipid in resistin gene expression and plasma resistin in young accumulation. Sex hormones are factors that may explain in obese Zucker rats possibly indicates a counter-regulatory part the gender-dependent BAT thermogenic response found role to maintain insulin sensitivity. in previous works.

T1:P1-079 T1:P1-080 PPARγ2 expression in response to cafeteria-diet feeding. Gender- and depot-specific effects. Causes of growth impairment in mice lacking GFRα2 Rodríguez E, Ribot J, Palou A Jari Rossi, Karl-Heinz Herzig, Vootele Võikar, Päivi H. Universitat de les Illes Balears, Palma de Mallorca, Spain Hiltunen, Mikael Segerstråle, and Matti S. Airaksinen

Aims: To investigate the effects of short-term cafeteria- (CAF-)diet feeding on the expression of adipogenic Abstract withdrawn transcription factors and their association with adiposity. Methods: 4-week-old male and female Wistar rats were fed CAF-diet or standard chow for 2 weeks. Parameters determined: PPARγ 2, PPARα, C/EBPα and ADD1 mRNAs in gonadal and inguinal white adipose tissues (gWAT and iWAT) and in brown adipose tissue (BAT), by RT-PCR; body weight; energy intake; depot weights; and serum parameters. Results: As compared to males and regardless of the diet, females had lower PPARγ 2 levels in subcutaneous depots (iWAT and BAT), that correlated with lower tissue weights. CAF-diet feeding resulted in increased body weight, adipose tissue weights and lipid serum levels. Changes of PPARγ 2 expression in response to CAF-diet feeding were restricted to females: in BAT, PPARγ 2 decreased, which could explain, at least in part, the previously described defective induction of BAT-thermogenesis in females; in gWAT, PPARγ 2 tended to increase and this correlated with a higher hypertrophy of the depot versus that in males. Conclusions: Our results implicate PPARγ 2 in gender- and depot-specific effects of CAF-diet on adipose tissues development and function.

International Journal of Obesity Abstracts S63 T1:P1-081 T1:P1-082 UCP3 protein regulation in human skeletal muscle fibre Endurance training increases whole muscle PPARα and types I, IIa and IIx is dependent on exercise intensity PGC-1 mRNA as well as and PPARα protein content in all A.P.Russell, E.Somm, M.Praz, A.Crettenand, O.Hartley fibre types of human subjects. A.Melotti, J-P.Giacobino, P.Muzzin, C.Gobelet, O.Dériaz A.P.Russell, J.Feilchenfeldt, M.Praz, A.Crettenand, Clinique romande de réadaptation SUVA, Sion, Switzerland C.Gobelet, JP.Giacobino, O.Dériaz Clinique romande de réadaptation SUVA, Sion, Switzerland Aims: To determine if human UCP3 fibre type regulation is influenced by exercise intensity. Aims: To determine the effect of endurance training on whole Methods: Whole muscle UCP3 mRNA and muscle fibre type muscle PPARα, β/δ, Y and PGC-1 gene expression as well protein content were quantified using real-time PCR and as PPARα protein content in the different muscle fibre types. immunofluorescence techniques, respectively before and Methods: Whole muscle PPARα, β/δ, Y and PPARγ after 6 weeks of endurance (ETr) and sprint (STr) running coactivator (PGC-1) gene expression and PPARα fibre type training. protein content were quantified using real-time PCR and Results: ETr down-regulated UCP3 mRNA and protein immunofluorescence techniques, respectively. levels, respectively by 65 and 30% as did STr by 50% and Results: Endurance training induced an increase in PGC-1 27%. ETr significantly reduced UCP3 protein content in type and PPARα mRNA expressions by 3.6 and 2.2-fold I, IIa and IIx muscle fibres by 54, 29 and 16%, as did STr by (p<0.01), respectively, with no change in the other PPAR 24, 31 and 26%. isoforms. Before training, PPARα protein was 1.5 fold greater Conclusions: ETr, when compared to STr, reduced in type I than in II type fibres (p<0.001). After training, significantly more and less, UCP3 expression in fibre types I PPARα protein content increase was greater, 3 and 1.5 fold, and IIx, respectively. This suggests that UCP3 regulation in respectively in type I than II fibres (p<0.01). Conclusions: the different fibre types is related to the relative recruitment Training increased whole muscle PGC-1 mRNA expression of the fibres, ie. the more a fibre is recruited, the more it and PPARα protein content in all fibre types. The increase in adapts to training by a decrease in its UCP3 expression. PGC-1 may be responsible, in part, for the increase in the Additionally, the more a fibre type depends on fatty acid β - percentage of oxidative fibres and mitochondrial content and oxidation and oxidative phosphorylation, the more it that of PPARα for the increase in fatty acid oxidative responds to ETr by a decrease in its UCP3 content. capacity, commonly observed after endurance training.

T1:P1-083 T1:P1-084 Zinc-α2-glycoprotein: In Vitro and In Vivo Lipolytic Effects of NO-synthase inhibition on food intake and Activity. leptin signalling in dietary obesity Russell ST, Zimmerman T, Domin B and Tisdale Sadler CJ, Wilding JPH MJ. Pharmaceutical Sciences Research Institute, Aston University of Liverpool, Liverpool, UK University, Birmingham, B4 7ET, UK. Aims: To assess the affects of systemic L-NAME on food Aims: Zinc-α2-glycoprotein (ZAG) was investigated for intake, body weight, hypothalamic NO and the involvement lipolytic activity in vitro and in vivo. of leptin signalling in chow-fed (C) and dietary-induced obese Methods: ZAG was isolated from human plasma and in vitro (DIO) rats. lipolytic activity was examined as release of glycerol from Methods: 24 male Wistar rats received chow or a palatable isolated murine adipocytes. ZAG was also incubated with diet for 9 weeks, then half received i.p. L-NAME (50 mg kg-1) SR59230A or Ro20-1724 to elucidate its mode of action. Ex- for 16 days. Hypothalamic NO content, phosphorylated JAK- breeder male mice were dosed i.v., i.p. or s.c. with 2, phosphorylated STAT-3, SOCS-3 and NPY, AGRP, endotoxin-free PBS or different dose levels of ZAG. POMC, SOCS-3 mRNAs were measured. Terminated mice were then analysed for body composition. Results: DIO rats were hyperleptinemic (4.57 ng ml-1 ± 0.31 Brown adipose tissue was removed and examined for vs 7.29 ng ml-1 ± 0.52; P<0.01). L-NAME reduced uncoupling protein 1(UCP1) expression by immunoblotting. cummulative food intake in C by 11% (5336 KJ ± 152 vs Results: ZAG was observed to be lipolytically active in 4771 KJ ± 70; P<0.05) and DIO by 16% (6401 KJ ± 269 vs isolated adipocytes (P<0.001). ZAG lipolytic activity was 5374 KJ ± 170; P<0.001); decreased NO in C (45.8 µM ± 4.8 inhibited by SR59230A and enhanced by Ro20-1724 vs 28.0 µM ± 2.7; P<0.05) and in DIO (49.1 µM ± 3.8 vs 37.6 (P<0.001). ZAG-treated mice exhibited a dose-dependent µM ± 3.0; P=0.054) and reduced body weight gain in DIO weight loss (p<0.001) with a decrease in total body fat ± ± content (P<0.01). Increased UCP1 suggested that there was (28.4 g 5.0 vs 3.0 g 9.5; P<0.05). Neither diet nor L- increased energy expenditure in the mice treated with ZAG. NAME altered any other proteins or mRNAs examined. Conclusions: ZAG appears to act via the β3-adrenoceptor Conclusions: The leptin signalling pathway is unaltered in the cAMP-mediated pathway with specific loss of body fat fed state in dietary obesity and is not involved in the greater without an effect on food and water intake, ZAG therefore suppression of food intake and bodyweight gain seen in DIO warrants further investigation as a anti-obesity therapy. rats during inhibition of NO synthesis.

International Journal of Obesity Abstracts S64 T1:P1-085 T1:P1-086 Production of neurotrophic factors in white adipocytes Gastric leptin levels are related to feeding conditions. and their autocrine action Sánchez J., Picó C., Oliver P., Palou A. Kimura K, Murashima I, Saito M Universitat de les Illes Balears, Palma de Mallorca, Spain Hokkaido University, Sapporo 060-0818, Japan Aims: The aim of this study was to determine whether there Aims: We have observed abundant mRNA expression of is a relationship between the rhythm of food intake and (NGF) not only in brown adipose tissue gastric leptin levels, and also to evaluate the effects but also in white adipose tissue (WAT), which receives much produced by fasting and a short-term period of food intake on less sympathetic innervation. The aims of this study are to the gastric leptin system. extend this preliminary finding and to explore possible role of Methods: 3-month-old male Wistar Rats housed at 22 °C NGF in WAT. with a 12 hours light/dark cycle and free access to food and Methods and Results: RT-PCR and ELISA analyses in rats water were used. Gastric and serum leptin levels were and mice revealed that WAT synthesized and secreted large analysed every 3 h over a 24 h period by ELISA. The amounts of NGF and other neurotrophins such as NT-3. response to 14 hours of starvation and to a short-term period Production of these neurotrophins was also confirmed in of food intake (20 minutes) was also studied. isolated white adipocytes and some cell lines including 3T3- Results: Gastric and serum leptin levels display a similar L1 and p53-KO adipocytes, being less after adipose diurnal rhythm. At the beginning of the light period, both differentiation. WAT and isolated adipocytes also expressed gastric and serum leptin levels are maximum and gradually significant levels of mRNA of neurotrophin receptors such as fall reaching a nadir before the onset of the dark period, with TrkA and p75. When primary cultured adipocytes were these parameters being related to the gastric content of food. stimulated by NGF, phosphorylation of ERK was increased in In addition, leptin secretion from the is stimulated by time- and dose-dependent manners, reflecting specific food intake, since a short food intake stimulus (20 min. re- activation of the MAP kinase cascade. feeding) following fasting produces a decrease in gastric Conclusions: Our results suggest that NGF produced from leptin levels and an increase in serum leptin. white adipocytes acts as an autocrine factor and may be Conclusion: Gastric and serum leptin levels are related to involved in the regulation of proliferation and/or differentiation feeding conditions. Furthermore, the fact that feeding of adipocytes. acutely stimulates gastric leptin secretion suggests a role of this protein in the short-term control of food intake.

T1:P1-087 T1:P1-088 Increased cardiac PPAR alpha expression in overweight/ THE IMPORTANCE OF PREPERITONEAL AND obese humans SUBCUTANEOUS FAT ON METABOLIC DISTURBANCES Schupp M, Pregla R, Kintscher U, Hetzer R, Sermez Y,Yaren A,Sabir N,Oflaz Z, De girmenciolu S. Unger Th, Regitz-Zagrosek V Pamukkale University Hospital,Obesity Study Group (OSG),Denizli-TURKEY Institute of Pharmacology, Charité, Berlin, Germany Aims: The aim of the study was to investigate wheter Aims: PPAR alpha is an important regulator of cardiac fatty preperitoneal and subcutaneous fat are important as well as visceral fat on metabolic disturbances in obese subjects. acid (FA) metabolism. To identify early changes of cardiac Methods: In this study 152 obese (BMI:37,4±6,2 kg/m² ) and 72 metabolism in obesity, we st udied PPAR alpha expression nonobese (BMI:24,3±4,1 kg/m² ) women were studied. Fasting in hearts from normal weight(NW) and overweight plasma glucose (FPG), postprandial glucose (PPG), total /obese(OW) individuals with normal cardiac function. cholesterol (T-C), triglyceride (TG), HDL-cholesterol (HDL-C), Methods: PPAR alpha mRNA expression was studied in left LDL-cholesterol (LDL-C), insulin (I), and uric acid (UA) were ventricular biopsies from OW (BMI: 25.3-33.1 kg/m2,n=7) and 2 determined. Visceral fat (VF),preperitoneal fat (PPF) and NW (BMI: 20.1-24.9 kg/m ,n=12) individuals with normal subcutaneous fat (SCF) were measured by ultrasonography. systolic cardiac function using real-time sybr green PCR. Results: FPG (p<0,001), PPG (P<0,01), I (p<0,001), T-C Results: Cardiac PPAR alpha mRNA was significantly (p<0,05), TG (P<0,001), ldl-c (P<0,05) and UA (p<0,001) were increased in the OW-group compared to the NW individuals higher in obese than nonobese. PPF,SCF and VF positively (1.4 ± 0.37-fold increase vs. NW, p<0.05). No correlation correlated with age, BMI and waist-to-hip ratio. PPF positively between cardiac PPAR alpha mRNA levels and age was correlated with I, and negatively correlated with HDL-C. There observed in the two groups. was a positive correlation between VF and biochemical Conclusions: This study demonstrates that OW humans with parameters including FPG, PPG, I, UA and TG .A negative normal systolic function have increased cardiac PPAR alpha correlation was found between VF and HDL-C. levels, which might be associated with enhanced cardiac FA- Conclusion: This study shows that VF is the most important oxidation. The increase in cardiac PPAR alpha might be an predictior in the evaluation of metabolic disturbances in obese initial step in the alteration of cardiac metabolism in obesity objects and also is found to be superior to SCF and PPF. resulting in cardiac dysfunction.

International Journal of Obesity Abstracts S65 T1:P1-089 T1:P1-090 Plasma adiponectin levels in relation to atherosclerotic A 3 dimensional-culture system with preadipocytes lipid profiles among school children in Taiwan autotransplanted in an animal model Shen MH, Chu NF, Wu DM Halbleib M, Aprath I, Hunt J, Skurk T, Rhodes N, de Luca C, Department of Public Health, NDMC, Taipei, Taiwan, ROC Hauner H. German Diabetes Research Institute, Düsseldorf, Germany Aims: To evaluate the association of plasma adiponectin levels and lipid profiles among school children. Aim: Objective of this project is the development of a soft Methods: We randomly selected 1,250 children (610 boys tissue-filler, using rat adipocyte precursor cells grown on 3 and 640 girls) aged 12-16 years in this study. We measured dimensional hyaluronic acid-based scaffolds. anthropometric variables, lifestyle factors and biochemical Methods: We implanted two different types of scaffolds parameters among these children. Plasma adiponectin levels loaded with autologous rat preadipocytes in the subscapular were measured by RIA method. We also measured lipid region of Sprague-Dawley rats. Preadipocytes were isolated profiles including serum total cholesterol (CHOL), triglyceride and proliferated in a monolayer culture before seeding on the (TG), HDL-cholesterol (HDL-C), apolipoprotein-A (Apo-A), scaffolds. After 3 days of in vitro differentiation scaffolds apolipoprotein-B (Apo-B), and lipoprotein(a) [Lp(a)] levels, were implanted. Scaffolds were explanted after 3, 8 and 12 and calculated LDL-C levels and total cholesterol to HDL-C weeks, respectively, and prepared for histological ratio (TCHR). examination. Results: Girls had higher adiponectin, CHOL, TG, HDL-C, Results: After removal, all scaffolds had a rather thick LDL-C, Apo-A1, Apo-B, and Lp (a) levels and lower BMI than capsule of connective tissue and showed new formation of boys. Plasma adiponectin levels were positively correlated vessels. Stained paraffin-sections also showed a massive in- with Apo-A1 and negatively correlated with TCHR in both growth of cells other than adipocytes inside the scaffolds. genders (all p <0.01). Only in few cases newly developed adipocytes were visible. Conclusions: Children with relatively higher plasma Conclusions: Preliminary histological observations suggest adiponectin levels had better lipid profiles such as higher that macrophages/monocytes represent a large proportion of HDL-C and apo-A1 levels and lower TCHR, however, it was in-growing cells on scaffolds in vivo. We therefore speculate only presented in boys but not girls. Further studies were that macrophage-derived factors may be responsible for the indicated to evaluate the possible mechanisms. lack of fat cell differentiation. This study was funded by the European Community (G5RD-CT-1999, Adipo- Regeneration).

T1:P1-091 T1:P1-092 Expression of mRNA for receptors in Cannabinoid 1 Receptor Inverse Agonist, AM251 hypothalami of fa/fa and fasted rats decreases body weight by decreased food intake Smith K, Medhurst A, Stocker C, Smart D, Arch J and increased fat utilisation University of Buckingham, Buckingham, and Strack, A, Stribling, S, Kuca, S, Shearman, L, Metzger, J, GlaxoSmithKline, Harlow UK MacIntyre, D.E. Merck Research Laboratories, Rahway, NJ, USA Aims: We have investigated whether the hypothalamic bombesin (BB) system responds to leptin signalling. Since Aims: Two-week administration of the CB1 inverse agonist, the natural agonist for the BB-3/BRS-3 receptor is not known, AM251, was examined to investigate the mechanisms by we have measured expression of mRNA for the three BB which CB1 receptor blockade decreases body weight. receptors. Y mRNA served as a control. Methods: Sprague Dawley rats, raised on a moderately high Methods: Hypothalami were taken from fed fa/fa, fed lean fat (32% kcal), high sucrose (25% kcal) diet (D12266B, (+/?), leptin-treated fed lean (1mg/kg ip, 10:00 and 17:00h Research Diets, NJ) were given AM251 (3 mg/kg, po) for 14 the previous day) and fasted lean Zucker rats. Gene days. Daily energy expenditure, food intake and body weight expression was measured using real time, PCR-based 5’- were measured. Before the start and end of the study, body nuclease (TaqMan) assays relative to three housekeeping composition was measured by DEXAscan analysis. genes: GAPDH, β-actin and cyclophilin. Results: AM251 treatment decreased final body weight by Results: Expression of BB-2 (GRP) receptor mRNA was 7%; fat, but not lean mass, was decreased. Daily food intake increased about twofold (P<0.01) relative to all three was suppressed ~ 34% for the first 7 days, after which a housekeeping genes in fa/fa and fasted rats compared to fed nonsignificant suppression of 10% was seen. Similarly, lean controls. Giving leptin to the fed rats had no effect. respiratory quotient (RQ) was decreased significantly There was no effect of treatment on expression of the BB-1 throughout the diurnal rhythm for the first 7 days, after which (NMB) or BB-3 receptors, though there was a trend to the RQ decrease was attenuated. O2 consumption was increased expression of the BB-3 receptor in fasted rats. unchanged. Expression of NPY mRNA was increased in fa/fa rats. Conclusions: Body weight loss in rats consequent to CB1 Conclusions: Expression of hypothalamic BB-2R mRNA is inverse agonist treatment results, at least in part, from elevated when leptin signalling is low, possibly as a response suppression of food intake and from increased fat utilization. to low levels of its anorexigenic ligand GRP.

International Journal of Obesity Abstracts S66 T1:P1-093 T1:P1-094

Predicting factor of vascular endothelial dysfunction in healthy The Serotonin 5-HT6 Receptor Antagonist BVT.5182 premenopausal obese women Reduces Body Weight of High Fat Diet-induced Mice Hee-Sun suh, MD., Kyung-Won Shim, MD., Jee-Hyun Kang, MD., Svartengren J, Öhman B, Edling N, Svensson M, Fhölenhag Yong-woo Park, MD.. Lee Sang-Hwa, MD., Lee Hong-Su, MD. Department of Family Medicine and medical research center, K, Axelsson-Lendin P, Klingström G, Larsson C, University of Ewha Womans College of Medicine, Seoul, Korea Biovitrum AB, Department of Biology, Stockholm, Sweden Aim: Vascular endothelial dysfunction(VED) plays a pivotal role in the pathogenesis of atherosclerosis and is associated with insulin resistance and visceral obesity. We examined the predicting factor of vascular Aims: We have investigated the role of the serotonin 5-HT6 endothelial dysfunction in healthy premenopausal obese women using receptor in feeding and body weight regulation using the pulse-wave analysis(PWA) combined with provocative pharmacological testing. selective 5-HT6 receptor antagonist BVT.5182 and the Methods: The subjects included 33 obese women (BMI 25), aged 20-45y, clinically established anti-obesity drug sibutramine. and 25 age-matched control subjects (BMI;18.5-22.9). All women were Methods: Acute effects were monitored for 21 hours in an sedentary(<1hr/wk of physical activity), non-smokers. We exclude those individuals who had type 2 diabetes melitus, hypertension, automated computer-assisted operant test cage system hyperlipidemia, cardiovascular disease, acute inflammatory disease. All using ob/ob mice. Long-term effects were monitored using tests were performed in the subjects in the folicular phase of their male C57BL/6J mice and SD rats fed a high fat diet (DIO) for menstruation cycle, which was within the first week after menstruation stops. Anthropometric measurements, metabolic variables, adipose 6 months and treated subcutaneously once daily or by tissue regional distribution, and endothelial function by performing pulse- infusion, respectively, for 14 or 29 days. Blood samples were wave analysis(PWA) combined with provocative pharmacological testing taken for clinical chemistry analysis, and epididymal fat was were done in the subjects. dissected. Results: Augmentation Index (AIx) fell significantly after the administration of salbutamol, which causes endothelium-dependent vasodilatation. Results: BVT.5182 potently decreased food intake but did This value was significantly reduced in obese women compared with not affect general motor activity and water intake at threshold controls (10.28 6.72% vs 17.2 6.84%, p=0.0003). Nitroglycerin that causes doses for effect on food intake in ob/ob mice (3 mg/kg sc). endothelium-independent vasodilatation did not result in significant changes. (30.86 9.67% vs 30.6 10.11%, p=0.9172). In our obese Repeated treatment with BVT.5182 reduced body weight by subjects, visceral adipose tissue area was the independently significant 9% and cumulative food intake by 11% in DIO mice (3 mg/kg predictor of vascular endothelial dysfunction regardless of hsCRP, Free sc). Sibutramine (10 mg/kg sc) decreased body weight with Fatty Acid, and HOMA score. ( =-0.1381, P=0.0038, Adj-R2=0.348) similar efficacy. BVT.5182 decreased serum leptin as well as Conclusion: Increased abdominal adiposity is a powerful independent predictor of VED in obese healthy women. Future studies are needed on epididymal fat. Similar effects were found in DIO rats. vascular endothelial function to determine the independent effects of Conclusion: These findings support that the serotonin 5-HT6 abdominal fat. receptor is involved in feeding and body weight regulation. Keywords: Visceral adipose tissue area, vascular endothelial dysfunction, C Augmentation Index. HsCRP(high-senstivity C-reactive protein).

T1:P1-095 T1:P1-096 Expression of human hormone-sensitive lipase in white Leptin and Bone: A Cross-Sectional Study in Obese and adipose tissue of transgenic mice Non-obese Men Tavernier G, Lucas S, Tiraby C, Mairal A, Langin D Morberg CM, Tetens I, Black E, Toubro S, Sorensen TI, INSERM U586, Toulouse, France Pedersen O, Astrup A Department of Human Nutrition, The Royal Veterinary and Aims: The purpose of the present work was to study the Agricultural University, Copenhagen, Denmark effect of a physiological increase of hormone-sensitive lipase (HSL) expression in vivo. Aims: This observational analysis was performed to explore Methods: Transgenic mice were produced with a 21 kb the relationship between serum leptin and BMD in obese and human genomic fragment encompassing the exons encoding non-obese men. the adipocyte form of HSL. Methods: The study population consisted of 327 non-obese Results: Human HSL mRNA and protein were readily men (controls) (BMI 26±3.7 kg/m2, age 49.9±6 y) and 285 detected in mouse white adipose tissue. The hydrolytic juvenile obese men (BMI 36±5.9 kg/m2, age 47.5±4 y). activities against triglyceride, diglyceride and cholesteryl Whole body DXA measured fat mass, lean mass and BMD. ester were increased in transgenic mouse adipose tissues. Physical activity and smoking were assessed by However, cAMP-inducible adipocyte lipolysis was lower in questionnaire. transgenic animals. On the B6CBA genetic background, Results: Leptin adjusted for fat mass (kg) was in inverse transgenic mice up to 14 weeks of age showed lower body correlated to BMD in the control group (r= -.125;P =.026). weight and fat mass. The phenotype was not observed in The inverse relation between BMD and serum leptin become older animals and in mice fed a high fat diet. On the OF1 stronger in both controls (r= -.186;P<.01) and juvenile obese genetic background, there was no difference in fat mass of (r= -.135;P<.05) when BMD was adjusted for body weight mice fed ad libitum. However, transgenic mice became (kg). In a multiple linear regression, fat mass (%) and lean leaner than their wild type littermates after a 4 day calorie mass (%) were both negatively associated to BMD. restriction. Conclusions: The study suggests that leptin, independent of Conclusions: The data show that overexpression of HSL, body size and composition, may have a negative influence despite increased lipase activity, does not lead to enhanced on BMD in men, and that increasing fat mass (%) do not lipolysis. protect obese men against osteoporosis.

International Journal of Obesity Abstracts S67 T1:P1-097 T1:P1-098 Real time PCR analysis of tissue factor gene expression Relationship between plasma free fatty acids, resting in rat adipose tissue and adipocyte primary culture energy expenditure and fat oxidation in lean and obese Keeley CRM, Rayner DV, Merryman-Simpson A, Trayhurn P. van Baak M.A. and S.L.H. Schiffelers. Dept Human Biology, Department of Medicine, University of Liverpool, Liverpool, NUTRIM, Universiteit Maastricht, Maastricht, The UK Netherlands

Aims: Tissue factor (TF) initiates the coagulation cascade in Aims. To study the relationship between changes in plasma the extrinsic clotting pathway, and is reported to be an free fatty acids (FFA) and changes in resting energy . The aim of this study was to examine TF gene expenditure (RMR) and fat oxidation in lean and obese. expression in white adipose tissue (WAT) and in adipocytes Methods. In different studies the plasma concentration of differentiated in primary culture using real time PCR. FFA was increased by various methods: administration of Methods: RNA was extracted from each of 14 tissues from different types of beta-adrenoceptor agonists or a triglyceride hooded Lister rats, adipose tissue of lean and ob/ob mice, emulsion plus heparin in lean and/or obese men. Changes in and from adipocytes before and after differentiation in plasma FFA concentrations and changes in RMR and fat primary culture. TF mRNA was quantitated using the oxidation, measured by indirect calorimetry (ventilated hood), Taqman system (Applied Biosystems). were studied. Results: TF mRNA was detected in all rat tissues examined. Results. A significant correlation between changes in plasma Higher levels of the mRNA were found in white (epididymal, FFA and RMR or fat oxidation was found in both lean and subcutaneous, perirenal, omental, mesenteric) and brown obese subjects across studies. No significant difference (interscapular) adipose tissues than in any other tissue. No between lean and obese was found. differences were found in TF mRNA level in adipose tissue of Conclusions. These results suggest that changes in lean and ob/ob mice, and neither fasting (24 h) nor cold availability of FFA are an important determinant of changes o exposure (24 h at 6 C) induced changes. In primary culture, in RMR and fat oxidation. Differences in response to stimuli TF mRNA level gradually increased post differentiation. that increase FFA availabilty between lean and obese Conclusions: It is concluded that WAT is a major site of TF subjects are due to differences in responsiveness of FFA gene expression in rodents and may be a key source of the release and not to a different relationship between the circulating protein. TF expression in WAT did not alter in change in plasma FFA and RMR or fat oxidation. obesity or when substrate flux and sympathetic activity were stimulated.

T1:P1-099 T1:P1-100 Effects of exercise and dietary intervention on human Lack of growth hormone, insulin and IGF-1 responses skeletal muscle metabolism in middle-aged IGT persons after oral load in obese subjects. Venojärvi M1, Puhke R, Hämäläinen H, Marniemi J, Rastas Vignati F, Muratori F, Di Sacco G. M, Hänninen O, Rusko H, Aunola S U.O. Endocrinologia Ospedale Niguarda, Milano, Italia 1University of Kuopio, Finland Aim: To evaluate the acute effect of oral essential amino Aims: The purpose of this study is to find out if middle-aged acids (EAA) load on anabolic hormones such as GH, IGF-1 obese people with IGT are able to improve their muscular and insulin (IRI), in a group of obese subjects. performance capacity, improve glucose metabolism and Methods. Six male obese subjects (mean age = 42.1±5.8 decrease oxidative stress in skeletal muscle by a 2-year years; mean BMI = 34.6±4.8 kg/m2) underwent blood samp- exercise and dietary intervention. ling for GH, IRI and glycemia before and every 30 min for 3 Methods: Study subjects were 22 persons from the Finnish hours after oral administration of 8g of an EAA mixture. Diabetes Prevention Study with IGT who volunteered to give Plasma IGF-1 was determined before and 24 hrs after the samples of biopsies from the vastus lateralis muscle. amino acids load. As control each patient underwent the Changes in glucose tolerance, the activities of muscle fibre same protocol, without EAA load, the day preceding the test. glycolytic/oxidative enzymes and markers of oxidative stress Results: Mean levels of GH, IRI, IGF-1 and glycemia did not were studied. The intervention consisted of (1) dietary advise change the day of the test in respect to the control day. and (2) strength & power and/or aerobic exercise training. Conclusions: The present study shows that oral Results: Exercise training increased LDH (lactate administration of EAA does not stimulates, in obese male dehydrogenase), LDH-1 (LDH isotype, heart) and CS (citrate subjects, GH and IGF-1 secretion and does not modify synthase) activities in vastus lateralis muscle. This, together significantly glycemia and IRI levels. with dietary advise decreased amount of the HO-1 (heme Control day EAA load oxygenase) and 4-HNE (4-hydroxy-2-nonenal) proteins in Basal Peak Basal Peak muscle tissue. Gly(mg/dL) 86.3±9.7 87.7±10.1 88.8±7.0 84±5.7 Conclusions: Exercise training improves both glycolytic and GH (ng/mL) 0.3±0.5 0.5±0.8 0.3±0.5 0.5±0.6 oxidative enzyme activities in vastus lateralis muscle and IRI (mU/mL) 15.5±9.9 11.9+6.8 17.9±8.2 11.9±7.4 may enhance protection against oxidative stress together IGF-1 (ng/mL) 164±172 - 171±167 185±165 with dietary advise in the middle-age persons with IGT.

International Journal of Obesity Abstracts S68 T1:P1-101 T1:P1-102 In vivo regulation of human skeletal muscle gene Effect of L-Carnitine on energy balance, UCP2 and 3mRNA. expression profile by epinephrine G Wittert, Z Holthouse, H Turnbull, Department of Medicine, Viguerie N, Barbe P, Poitou C, Thalamas C, Barsch G, University of Adelaide, Australia 5000. Clément K, Langin D INSERM U586, Toulouse, France Aim: To determine the effects of increasing CPT1 activity with L- carnitine on energy balance in the marsupial S. crassicaudata. Aims: Our goal was to determine the changes in human Methods: Exp 1. Lab diet (labd) (20% fat, 1.01 kcal/g) or skeletal muscle gene expression induced by catecholamines. mealworm (mw) (30% fat, 2.99 kcal/g) fed females (n=6/group) Methods: Nine young healthy males received 0.04µg/min/kg were injected IP with L-carnitine (50 - 200 mg/kg) or vehicle at of epinephrine for 6 hours. Biopsies of vastus lateralis the onset of the dark phase, after a 24-hour fast, refed with muscle were performed before and at the end of the baseline diet and food intake measured. Exp 2. L-carnitine (200 mg/kg) or vehicle injected IP daily for 21 days in labd or mw fed treatment. Fluorescent-labeled cDNA prepared from animals (n=8/group). Food intake, weight, tail width (reflects fat amplified total RNA were hybridized to microarrays stores) and physical activity were measured daily, glucose containing 49000 human cDNAs. tolerance at baseline and day 14. On day 21 plasma FFA and Results: Plasma epinephrine levels increased from 23±4 to glucose and UCP2/3 and leptin mRNA were measured. 787±68 pg/ml (p<0.02). Resting metabolic rate increased Results: Exp1. Food intake increased in mw fed animals in from 92±6 to 109±3 J/min/kg lean body mass (p<0.05). response to L-carnitine, 100 and 200 mg/kg (p=0.047 and Statistical analysis of microarray data showed upregulation p=0.014). Exp2. L-carnitine didn’t effect weight, activity, plasma for 1218 mRNAs and downregulation for 642 mRNAs. Data glucose, glucose tolerance or leptin mRNA in either labd or mw from a subset of genes are under validation using real-time fed animals, but induced a decrease in tail width in labd PCR. Classification into functional categories revealed new (p=0.03) and mw (p=0.047) fed animals. Food intake increased sets of genes regulated by epinephrine. only in those fed mw (p=0.03). Plasma FFA levels decreased Conclusions: Epinephrine regulates a large repertoire of only in those fed labd (p=0.037). UCP2 mRNA increased in fat genes in vivo in human skeletal muscle. Characterization of (p=0.05) and liver (p=0.002) in response to L–carnitine in labd unknown patterns of variations improves our understanding fed animals but only in fat (42% p =0.05) in those fed mw. There of the effect of catecholamines on skeletal muscle and were no changes in gastrocnemius UCP2 and UCP3mRNA. provide new insights in the regulation of energy expenditure. Conclusion: The effect of mitochondrial transfer of FFA’s on energy balance and UCP2mRNA is diet and tissue-dependant.

T1:P1-103 T1:P1-104 Expression of Class III facilitative glucose transporter Role in vivo of angiotensin type 2 receptor on adipose protein (GLUT) family members in mouse adipose tissue tissue development and metabolism. Wood IS, Hunter, L, Trayhurn, P Yvan L, Even P, André J, Ménéton P, Quignard-Boulangé A. Dept. of Medicine, University of Liverpool, Liverpool, UK INSERM U465, IFR 58, Paris, France

Aims: Glucose is central to the function of adipose tissue and Aims: Angiotensin type 2 receptor (AT2) has been involved its transport into adipocytes is considered to occur through in vitro in the stimulation of adipose differentiation and the the GLUT1 and GLUT4 (insulin sensitive) transporters. activation of the lipogenesis. In order to understand the role Recently, the family of facilitative transporters (GLUTs) in vivo of AT2 in adipose tissue development, we used mice has expanded to include 8 new members. Of these, GLUT10 lacking AT2 receptor gene (AT2 KO). has been found in a type II diabetes-linked region of human Methods: 13-week-old wild type (WT) and AT2 KO mice were 20 and GLUT12 expression has been reported used. Mice were placed during 24 hours in a metabolic in human adipose tissue. In this study we have investigated chamber to determine basal and resting metabolic rate and the expression profile of these novel glucose transporters in to record food intake and spontaneous activity. After mouse adipose tissue. sacrifice, the cellularity (size and number of adipocytes) of Methods: RT-PCR was used to determine the expression of epididymal adipose tissue as well as the expression (activity GLUT10 and GLUT12 in mouse adipose tissue and in the and mRNA) of fatty acid synthase (FAS) were measured. 3T3-L1 adipocyte clonal cell line. Results : During development up to 13 weeks of age, there Results: Expression of both GLUT10 and GLUT12 was found was no difference in growth rate between WT and AT2 KO in mouse adipose tissue (gonadal, perirenal, subcutaneous) mice. Basal metabolic rate was similar in both groups. A and occurred in adipocytes and the stromal vascular fraction. significant decrease in spontaneous activity and resting GLUT10 was also expressed in 3T3-L1 cells, before and metabolic rate was observed in AT2 KO mice. Adipose tissue after differentiation. The mouse GLUT12 cDNA sequence of KO AT2 mice was characterized by a cellular hypotrophy was found to be very similar to its human homologue. associated with an increased cell number as compared to Conclusions: The addition of GLUT10 and GLUT12 to the the controls. FAS activity and its expression were decreased existing list of GLUTs expressed in adipose tissue suggests by 70%. that the transport of glucose into adipocytes and its control is Conclusion : This study demonstrates in vivo the importance more complex than previously considered. of AT2 receptor in adipose tissue enlargement by controlling size and number of adipose cells.

International Journal of Obesity Abstracts S69 T1:P1-105 T1:P1-106 Effect of weight loss on serum concentrations of ghrelin Cerebrospinal fluid ghrelin is negatively associated with and growth hormone body mass index Zahorska-Markiewicz B., Janowska J., Olszanecka- Tritos NA, Kokkinos A, Lampadariou E, Alexiou E, Glinianowicz M., Katsilambros N, Maratos-Flier E Department of Pathophysiology, Silesian University School of First Propaedeutic Department of Internal Medicine, Medicine, 40 – 752 Katowice, Poland University of Athens Medical School, Athens, Greece

Aims: Determine whether weight loss treatment modulates Aims: Several studies suggest that ghrelin has a role in serum concentrations of ghrelin and growth hormone. appetite regulation. Our aim was to further characterize the Methods: 25 obese women ( age 40,7 ± 11,9 y, BMI before role of ghrelin in the regulation of body adiposity by 36,8 ± 5,3 kg/m2 after 33,6 ± 5,2 kg/m2 ) were examined investigating the association between cerebrospinal fluid before and after 3 months weight reducing treatment, (CSF) ghrelin levels and body mass index (BMI). consisting of 1000 kcal/day diet and physical exercises. Methods: We enrolled 19 adults; 4 obese, 7 overweight and 8 Serum concentration of ghrelin was measured by ELISA. lean subjects, who underwent spinal anesthesia during Serum concentration of growth hormone was measured by surgery for non-malignant conditions and had no history of RIA heart, liver, or malignant disease. There was no Results: *p< 0,01; **p< 0,0005; ***p<0,0001 difference in age, gender distribution and plasma glucose Weight BMI Body fat Ghrelin GH levels between the 3 subject groups. (kg ) ( kg/m2 ) ( % ) pg/ml (µIU/ml ) Results: We found a negative association between fasting before 96,5±16,5 36,8±5,3 43,5±8,5 66,3±13,7 1,3±2,2 CSF ghrelin and BMI (r= -0.48, p=0.035) and a trend towards *** *** * * ** lower fasting CSF ghrelin in the obese (obese: 16.2±2.0 after 87,7 ± 15,2 33,6±5,2 40,9±6,9 73,7±14,8 4,4±4,5 pg/ml, overweight: 18.7±1.0 pg/ml, lean; 19.4±0.5 pg/ml, Significant positive correlation of ghrelin with GH ( r =0,42, p=0.063 for the difference between lean and obese subjects). p<0,05) before treatment was found. Conclusions: Our study is the first to demonstrate a negative Conclusions: Increased serum concentration of ghrelin and association between fasting CSF ghrelin levels and BMI. It is growth hormone after weight loss, may suggest that ghrelin not clear whether CSF ghrelin originates from the systemic is one of factors stimulating secretion GH after body mass circulation or the brain. However, our data suggest that CSF reduction. ghrelin has a role in the regulation of body adiposity.

International Journal of Obesity