2007 Annual Report 1a.Objectives (from AD-416) 1) Identify a number of genes that affect the expression of childhood obesity in Hispanic children, investigate strong positional candidate obesity-related genes, and test if weight changes and metabolic, hormonal and immunologic responses to weight changes are dependent upon genotype;. 2)Establish a reference model of body composition in children;. 3)Identify barriers and facilitators for the physical activity component of the 2005 Dietary Guidelines for Americans and relate to obesity risk in urban, African-and Mexican-American children and families;. 4)Determine the contribution of leukocytes to the composition and function of adipose tissue, investigate the influence of dietary factors on the composition of leukocytes within adipose tissue and liver, and explore how obesity can both reduce host resistance and enhance inflammatory tissue injury;. 5)To determine whether a 12-week exercise program without intent to weight loss would increase insulin sensitivity and reduce insulin secretion and glucose production from gluconeogenesis in obese adolescents, and if so, whether these changes are associated with a decrease in intramyocellular and intrahepatic fat content; and. 6)To develop a greater understanding that altered sleep patterns associated with our "24-hour" lifestyle may contribute to the accumulation of body fat, and that such altered sleep patterns may ultimately represent alterations in both the central and peripheral circadian clock mechanisms. 1b.Approach (from AD-416) 1) A systemic genomic scan and follow-up fine mapping and sequencing of positional candidate genes will be performed on 300 overweight Hispanic children and their biological parents and siblings with respect to adiposity, the regulation of food intake, energy expenditure and energy partitioning before and after weight loss.. 2)A 5-level body composition model will be determined from multiple-method body composition assessments in 1500 multi-ethnic children. The predictive accuracy of the model for the individual will be verified by longitudinal restudy.. 3)One hundred10-15 year-old children/adolescents who are at risk for obesity (BMI> 85th percentile and < 97.5th percentile for age and gender and parental BMI >25) will be randomly assigned to the 12 week intervention or self help group.. 4)Animal models (murine) and human tissue will be used to characterize myeloid and lymphoid cells within adipose tissue and changes that occur in mice fed high fat diets. Phenotypic markers and in situ hybridization will be used to characterize the cell types, ultrastructural studies, and confocal microscopy will define the position of these cells in relationship to other structural components of the adipose tissue, and tissue fractionation techniques will be used to isolate these cells for function studies in vitro. Cell lines (e.g., RAW cells and 3T3 cells) will be used to study cytokine, chemokine and hormone release, adipocyte differentiation and lipid metabolism. Contributions of the immune cells to lipid metabolism in vivo will be sought in mice with disrupted or activated (e.g., with endotoxin) immune cell functions.. 5)Insulin sensitivity and secretion will be studied using stable isotopes, and intramyocellular and intrahepatic fat content will be determined using MRI.. 6)A 1-y after-school physical activity intervention will be conducted in 900 3rd- and 4th-grade Hispanic students from 12 elementary schools (6 intervention schools and 6 control schools). Dual-energy x-ray absorptiometry, doubly-labeled water method, and accelerometers will be used to assess changes in body composition, energy expenditure and level of physical activity, respectively.. 7)Through employing both hypothesis-generating and hypothesis-testing approaches to ascribe roles for the circadian clock within the adipocyte will increase our understanding of altered sleep patterns and how they may contribute to the accumulation of body fat. 4.Accomplishments Providing Preliminary Results on the Effects of Exercise on Insulin Sensitivity and Fat Distribution in Adolescents: Additional information is needed on the impact of exercise programs for obese adolescents. Children's Nutrition Research Center researchers demonstrated that controlled exercise training without weight loss resulted in increased fitness, reduced visceral fat accumulation in the abdomen and surrounding vital organs, and improved insulin sensitivity in obese adolescents. In addition, in obese adolescents with fatty liver, the exercise program reduced the amount of liver fat substantially (~ 40%). These preliminary results indicate that a moderate exercise program that can be accomplished by sedentary obese adolescents without losing weight can significantly improve their metabolic parameters involved in obesity-related disease. Findings from this study can be used in schools and obesity clinics as a strategy to improve insulin sensitivity and potentially delay or reduce the risk of type 2 diabetes mellitus. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 5) T cell Accumulation in Adipose Tissue: The molecular mechanism responsible for the inflammatory response in diet-induced obesity is not completely understood. Children's Nutrition Research Center researchers documented for the first time that white blood cells accumulated in fat tissue of obese individuals. Additionally, researchers identified a protein that is capable of attracting these cells to fat tissue. Researchers analyzed cell types and proteins in adipose tissue in an animal model (mice fed a diet high in saturated fatty acids) and in human biopsy tissue (humans with morbid obesity and the metabolic syndrome). The outcome of this work will be very important since it is the first step in analyzing the contributions of the immune system in adipose tissue, and it provides another marker for the early detection of systemic inflammation in obesity. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 4) ICAM-1 and Activated Macrophages in Adipose Tissue: Identifying the molecular mechanism responsible for the inflammatory response in diet-induced obesity is needed. Scientists at the Children's Nutrition Research Center documented activated macrophages in fat tissue in diet-induced obesity, and identified an important marker for inflammation in the tissue. TResearchers analyzed adipose tissue and blood in an animal model (mice fed a diet rich in saturated fatty acids). The impact of this work is very significant since it represents the recognition of a second white blood cell type in fat inflammation and the occurrence of a key marker. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 4) Identification of Clock-Regulated Genes within the Adipocyte: Through our experiments, Children's Nutrition Research Center scientists identified the clock-regulated genes within the fat cell. Genes within pathways related to transport, transcription, development, cell cycle, metabolism, cell division, and in particular, lipid biosynthesis were found to be altered in adipocyte-specific dominant negative CLOCK mutant (ACM) mice. Many of these genes were not known to be regulated by the circadian clock (internal body clock) prior to these experiments. Genes related to lipogenesis, or the production of lipids, were all elevated at the transition from sleeping to waking in the transgenic animals, indicating increased potential for fat formation during this time period. These findings suggest that the nutrient makeup of the diet and timing of feeding may both impact and/or be impacted by disruptions in normal patterns of circadian rhythms. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 6) Characterization of an Adipocyte-Specific CLOCK Mutant Animal Model: Scientists at the Children's Nutrition Research Center, Houston, TX ,established a transgenic colony of adipocyte-specific dominant negative CLOCK mutant (ACM) mice. The ACM animal model is the first model with a specific disruption of the circadian clock only within the fat cell. While other animal models with a global loss of the circadian clock have been reported to develop obesity and associated co-morbidities, both behavior and metabolism are altered in these ubiquitously disrupted animals, and it is not known whether the development of obesity in these animals is due to behavioral changes or direct alterations by the circadian clock. Through the use of the ACM animal model, we will be able to understand the means by which alterations in circadian patterns of metabolism and gene expression can lead to obesity. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 6) Assessing DXA Software for Improved Accuracy: In order to accurately calculate body composition measurements in children, additional assessment of the dual energy X-ray absorptiometry (DXA) tool and related software is needed. Our independent assessment of the newest software changes for DXA scans in children have shown that body fat values are increased for all children weighing less than 40 kg, compared with older software once used. Therefore, children examined in 2007 could appear to have increased body fatness compared with children of the same weight from only a few years ago. This software effect may contribute to a false perception that children are continuing to get fatter, which is important to recognize in nutrition-related research. Most importantly, a more accurate tool can be utilized for future research in children. [NP107, Component 6 Prevention of Obesity & Disease] (CNRC Project 2) 6.Technology Transfer Number of non-peer reviewed presentations and proceedings 28 Number of newspaper articles and other presentations for non-science audiences 184 Review Publications Ellis, K.J., Yao, M., Shypailo, R.J., Urlando, A., Wong, W.W., Heird, W.C. 2007. 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The intrinsic circadian clock within the cardiomyocyte. American Journal of Physiology - Heart and Circulatory Physiology. 289:H1530-H1541. Fornage, M., Lee, C.R., Doris, P.A., Bray, M.S., Heiss, G., Zeldin, D.C., Boerwinkle, E. 2005. The soluble epoxide hydrolase gene harbors sequence variations associated with susceptibility to and protection from incident ischemic stroke. Human Molecular Genetics. 14(19):2829-2837. Brake, D.K., Smith, E.O., Mersmann, H.J., Smith, C.W., Robker, R.L. 2006. ICAM-1 expression in adipose tissue: Effects of diet-induced obesity in mice. American Journal of Physiology Cell Physiology. 291(6):C1232-1239. Tyler, C., Johnston, C.A., Fullerton, G., Foreyt, J.P. 2007. Reduced quality of life in very overweight Mexican American adolescents. Journal of Adolescent Health. 40(4):366-368. Johnston, C.A., Steele, R.G. 2007. Treatment of pediatric overweight: an examination of feasibility and effectiveness in an applied clinical setting. Journal of Pediatric Psychology. 32(1):106-110. Foreyt, J.P. 2006. The role of lifestyle modification in dysmetabolic syndrome management. In: Bantle, J.P., Slama, G., editors. Nutritional Management of Diabetes Mellitus and Dysmetabolic Syndrome. Basel: Karger Publishers. p. 197-206. Espeland, M.A., Dotson, K., Jaramillo, S.A., Kahn, S.E., Harrison, B., Montez, M., Foreyt, J.P., Montgomery, B., Knowler, W.C. 2006. Consent for genetics studies among clinical trial participants: Findings from Action for Health in Diabetes (Look AHEAD). Clinical Trials. 3(5):443-456. Butte, N.F., Cai, G., Cole, S.A., Wilson, T.A., Fisher, J.O., Zakeri, I.F., Ellis, K.J., Comuzzie, A.G. 2007. Metabolic and behavioral predictors of weight gain in Hispanic children: The Viva la Familia Study. American Journal of Clinical Nutrition. 85(6):1478-1485. Bray, M.S., Young, M.E. 2006. Circadian rhythms in the development of obesity: Potential role for the circadian clock within the adipocyte. Obesity Reviews. 8(2):169-181. Wu, H., Ghosh, S., Perrard, X.D., Feng, L., Garcia, G.E., Perrard, J.L., Sweeney, J.F., Peterson, L.E., Chan, L., Smith, C.W., Ballantyne, C.M. 2007. T-cell accumulation and regulated on activation, normal T cell expressed and secreted upregulation in adipose tissue in obesity. Circulation. 115(80):1029-1038. Tejero, M.E., Cai, G., Goring, H.H.H., Diego, V., Cole, S.A., Bacino, C.A., Butte, N.F., Comuzzie, A.G. 2007. Linkage analysis of circulating levels of adiponectin in Hispanic children. International Journal of Obesity. 31(3):535-542. Ferdinands, J.M., Mannino, D.M., Gwinn, M.L., Bray, M.S. 2007. ADRB2 Arg16Gly polymorphism, lung function, and mortality: Results from the atherosclerosis risk in communities study. PLoS One. 2(3):e289. Cai, G., Cole, S.A., Haack, K., Butte, N.F., Comuzzie, A.G. 2007. Bivariate linkage confirms genetic contribution to fetal origins of childhood growth and cardiovascular disease risk in Hispanic children. 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Seminara, D., Khoury, M.J., O'Brien, T.R., Manolio, T., Gwinn, M.L., Little, J., Higgins, J.T., Bernstein, J.L., Boffetta, P., Bondy, M., Bray, M.S., Brenchley, P.E., Buffler, P.A., Casas, J.P., Chokkalingam, A.P., Danesh, J., Smith, G.D., Dolan, S., Duncan, R., Gruis, N.A., Hashibe, M., Hunter, D., Jarvelin, M., Malmer, B., Maraganore, D.M., Newton-Bishop, J.A., Riboli, E., Salanti, G., Taioli, E., Timpson, N., Uitterlinden, A.G., Vineis, P., Vareham, N., Winn, D.M., Zimmern, R., Ioannidis, J.P.A., for the Human Genome Epidemiology Network and the Network of Investigator Networks. 2007. The emergence of networks in human genome epidemiology: Challenges and opportunities. Epidemiology. 18(1):1-8. Poston, W.S.C., Haddock, C.K., Pinkston, M.M., Pace, P., Reeves, R.S., Karakoc, N., Jones, P., Foreyt, J.P. 2006. Evaluation of a primary care-oriented brief counselling intervention for obesity with and without orlistat. Journal of Internal Medicine. 260:388-398. Voruganti, V.S., Goring, H.H., Diego, V.P., Cai, G., Mehta, N.R., Haack, K., Cole, S.A., Butte, N.F., Comuzzie, A.G. 2007. Genome-wide scan for serum ghrelin detects linkage on chromosome 1p36 in Hispanic children: Results from the Viva la Familia study. Pediatric Research. 62(4):445-450. Rankinen, T., Bray, M.S., Hagberg, J.M., Perusse, L., Roth, S.M., Wolfarth, B., Bourchard, C. 2006. The human gene map for performance and health-related fitness phenotypes: The 2005 update. Medicine and Science in Sports and Exercise. 38(11):1863-1888. Lee, C.R., North, K.E., Bray, M.S., Avery, C.L., Mosher, M.J., Couper, D.J., Coresh, J., Folsom, A.R., Boerwinkle, E., Heiss, G., Zeldin, D.C. 2006. NOS3 polymorphisms, cigarette smoking, and cardiovascular disease risk: The atherosclerosis risk in communities study. Pharmacogenetics and Genomics. 16(12):891-899. White, J.S., Foreyt, J. 2006. Ten myths about high-fructose corn syrup. Food Technology. 60(10):96. Ellis, K.J., Grund, B., Visnegarwala, F., Thackeray, L., Miller, C.G., Chesson, C.E., El-Sadr, W., Carr, A. 2007. Visceral and subcutaneous adiposity measurements in adults: Influence of measurement site. Obesity. 15:1441-1447. Motil, K.J. 2006. Necrotizing enterocolitis. In: McMillian, J.A., Feigin, R.D., DeAngelis, C.D., Jones, M.D., editors. Oski's Pediatrics: Principles and Practice of Pediatrics. 4th edition. Philadelphia, PA: Lippincott Williams & Wilkins. p. 389-397. Quiros-Tejeira, R.E., Rivera, C.A., Ziba, T.T., Mehta, N., Smith, C.W., Butte, N.F. 2007. Risk for nonalcoholic fatty liver disease in Hispanic Youth with BMI > or = 95th percentile. Journal of Pediatric Gastroenterology and Nutrition. 44(2):228-236. Oyafemi, O.M., Ellis, K.J., Griffin, I.J., Abrams, S.A. 2005. Bone mineral status asessment by ultrasound in preterm infants. International Journal of Body Composition Research. 3(4):141-145. Shypailo, R.J., Ellis, K.J. 2005. 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