2006 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? This CRIS unit evaluates childhood obesity, specifically the regulation of energy balance and body composition. It is composed of six individual research projects;. 1)genetic and environmental factors contributing to childhood obesity;. 2)biological diversity of human growth: body composition references standards for children;. 3)prevention of overweight in children;. 4)nutrition influences on innate immunity;. 5)effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents; and. 6)circadian clocks in adipose biology and obesity. This research contributes to the goals of NP107, Human Nutrition, specifically "Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease". It address ARS Strategic Plan Goal #4 Improve the Nation's Health, specifically objective 4.1: Promote Healthier Individual Food Choices and Lifestyles and Prevent Obesity; Improve Human Health by Better Understanding the Nutrient Requirements of Individuals and the Nutritional Value of Foods; Determine Food Consumption Patterns of Americans. Project 1: Genetic and environmental factors contributing to childhood obesity Childhood obesity in the United States has dramatically increased in the past decade according to the latest National Health and Nutrition Examination Survey (NHANES), particularly among Mexican-American children. Childhood obesity is associated with hypertension, hyperinsulinemia, steatohepatitis, and alterations in lipoprotein metabolism. Childhood obesity not only presents serious medical and psychosocial consequences to the child, but also predisposes to adult obesity and its complications. Obesity is a complex multi-factorial disease. The current surge in obesity in the United States is due to an interaction between a genetic predisposition toward efficient energy storage and a permissive environment of readily available food and sedentary behaviors. Despite the high prevalence of obesity among Hispanic children, the etiology underlying the heightened susceptibility to childhood obesity in Hispanic populations has not been investigated. Researchers at the Children's Nutrition Research Center, Houston, TX, will assess the relative contributions of genetic and environmental (diet and physical activity) factors influencing childhood obesity and identify several positional candidate genes throughout the genome that are linked with obesity-related phenotypes in Hispanic children. The main objective of this project is to identify a number of genes that affect the expression of childhood obesity in Hispanic children. Although this project focuses on Hispanic children, all American children should benefit from prevention and/or management strategies aimed at childhood obesity. Scientists and clinicians will benefit from an increased understanding of the genetic and environmental causes of childhood obesity. USDA food and nutrition programs may adopt strategies to prevent childhood obesity based on this study's findings. Several genes that are linked with the quantitative variation in obesity-related phenotypes in children will be identified. Genetic polymorphisms will be identified that can be used to screen children at increased risk of developing obesity before excessive weight gain occurs. In addition, by measuring phenotypes such as those related to adiposity, the regulation of food intake, energy expenditure, and energy partitioning during growth, we will identify metabolic markers for early screening of "at risk" individuals. Effective strategies for the prevention and management of childhood obesity will be developed with the goal of reducing the staggering medical and societal costs associated with childhood obesity. Project 2: Biological diversity of human growth: body composition reference standards for children Obesity is defined as excess stores of body fat, and is a major health problem in the United States. The prevalence of childhood obesity has continued to start at younger ages and tracks into adulthood. This pattern is seen in all ethnic groups in the United States, with the largest increases occurring in minority populations. Different indices of fatness based on weight and height have been proposed, but these can be inaccurate for the individual. There are only a few analytical techniques that can accurately measure the amount of fat in the human body. If we are to better assess the impact of changes in the environment, such as modification of diets and/or physical (in)activity, improved measures of body fatness for the individual are needed, especially for growing children. As yet, the range of "normal" body fatness for children has not been scientifically established. Our lab continues to obtain precise body composition measurements in children at all ages (birth to 18 years) using different analytical techniques. Longitudinal changes in a subgroup of children have been followed over a 10-year period. The percentile distributions for body composition (amount of fat, bone, muscle, water) in the general pediatric population are being established. This information will be used to provide the most accurate estimates for the normal growth patterns for age, gender, and ethnicity, and establish the upper limits for normal adiposity for children. This information continues to provide a contemporary and unique pediatric reference that is available to health professionals to help establish more appropriate nutritional and activity guidelines for achieving acceptable growth without accumulation of excess body fat. Nutrition scientists, dietitians, exercise specialists, and pediatricians can use this information when assessing the nutritional status of children. Project 3: Prevention of overweight in children Our goal is to develop a program that promotes sustained weight management in children and adolescents, especially one that is effective for use with a Hispanic population. Our researchers will determine treatment effects on other health outcomes as well as psychosocial functioning and will test our objectives by conducting a randomized clinical trial in which children are assigned to either a behavioral, family-based weight management program led by an instructor or a self-help led by a parent. This research project will have a number of implications, especially considering there are no studies indicating they are effective weight management programs for Hispanic children. Additionally, research is lacking in terms of the health benefits achieved through weight loss and the sustainability of these changes long term. We will be able to examine beneficial health outcomes related to weight loss in children, a central mission of ARS National Program 107 – Human Nutrition. Project 4: Nutritional influences on innate immunity The major problem being addressed in this research project is the effect of high fat and high sucrose diets on adipose tissue and the liver. Obesity is a major health problem, and there is growing evidence that high fat and high sucrose diets cause systemic inflammation that contributes to many of the complications of obesity such as arterial disease. In addition, such diets are associated with fatty changes in the liver often with liver cell injury leading to cirrhosis. We are working on the mechanisms by which obesity causes inflammatory mediators to be generated in adipose tissue and how obesity causes liver injury. Our approach is to use animals of dietary obesity, cell and molecular biology. The work is important to those attempting to treat and prevent the complications of obesity. Project 5: Effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents Over the past 30 years the prevalence of overweight children (6-19 years) in the USA (BMI>95th percentile) has increased 3-fold to reach 15% in 2000, with an even higher incidence in Hispanic and African American adolescents (24% in both). Concomitant with the increase in childhood overweight, the incidence of type 2 diabetes in children (0-19 years) has increased 4-fold with predominance for African American and Hispanic children. We have demonstrated that healthy obese adolescents are significantly insulin resistant regardless of diet, but maintain normoglycemia, normolipidemia, and appropriate substrate oxidation rates by increasing their insulin sensitivity more than 2-fold as compared to their lean counterparts. This will, however, place increased demands on their pancreatic Beta-cells, thus increasing their risk of type 2 diabetes. These results emphasize the importance of preventing obesity and treating insulin resistance. It is well-known that achieving and maintaining weight loss is very difficult, particularly in adolescents. Therefore, the primary objectives of this proposal are to determine in healthy adolescents whether a 12-week exercise program improves insulin sensitivity and glucose metabolism; and whether exercise-induced metabolic effects are correlated with changes in intramyocellular, visceral, and/or hepatic lipid content and, whether ethnicity, gender and obesity have any impact on metabolic effects of exercise. To address these questions, 96 adolescents will be studied on two occasions (just prior to the start of a 12-week exercise program and during the last week of exercise) using state of the art stable isotope and magnetic resonance imaging (MRI) techniques. The information provided by these studies will improve our understanding of the metabolic effects of exercise and obesity in adolescents and will help in identifying factors that may increase the risk of type 2 diabetes, and designing new treatment strategies to prevent or delay the development of type 2 diabetes in adolescents. The knowledge that is likely to be gained by these studies will benefit adolescents, particularly from minority groups, who are at high risk of becoming overweight and developing insulin resistance and type 2 diabetes in that the information will be used in developing strategies to improve insulin sensitivity, thus preventing and/or delaying type 2 diabetes. This research will provide detailed information on the effects of obesity and exercise on visceral, hepatic and intramyocellular lipid content, glucose and lipid metabolism, and insulin sensitivity and secretion in adolescents. This information is crucial in defining treatment strategies to improve insulin sensitivity and glucose metabolism in adolescents, thus reducing their risk of or delaying the onset of type 2 diabetes. This information from this research will be used by adolescents in general and those who are overweight in particular and their parents; health care professionals taking care of obese adolescents and young patients with type 2 diabetes; policy groups designing nutritional and lifestyle guidelines for children and adolescents; school professionals, e.g., principals, school nurses, Physical Education teachers, committees planning educational programs; and organizations responsible for after school activities. Project 6: Circadian clocks in adipose biology and obesity More than two-thirds of the adults in the U.S. and more than 9 million children age 6-16 yrs are currently considered overweight or obese. Obesity is the second leading cause of preventable death in the United States behind smoking, and obesity is a major determinant of many other diseases, including type 2 diabetes, coronary heart disease, stroke, several types of cancer, gallbladder disease, musculoskeletal disorders, and respiratory problems. Abdominal obesity, in particular, is associated with insulin resistance and an atherogenic phenotype characterized by hypertension, hyperlipidemia, hyperglycemia, and a prothrombotic. Because obesity has a multifactorial etiology, the strategies and methods for its treatment are equally numerous and varied. An estimated $30 to $50 billion is spent yearly on unsuccessful efforts to lose weight, and the success rate of most long-term intervention programs for weight loss is dismal, illustrating the fact that obesity is both difficult to alleviate and, even today, not well understood. Recent reports have suggested that altered sleep patterns related to our "24-hour" lifestyle are associated with increased body fat and obesity, although the physiologic mechanisms that promote the accumulation of adipose tissue are not known. This project is designed to investigate the impact of disruption of normal intrinsic circadian rhythms, a potentially important contributor to the recent rise in human obesity, on the lipogenic and/or lipolytic function of the adipocyte. This research is relevant for physicians, interventionists, and prevention specialists, in recognizing lifestyle and physiologic factors that may have a profound influence both on response to obesity treatments as well as prevention of obesity onset by increasing our understanding of potential mechanisms that exacerbate the susceptibility for accumulating body fat. 2.List by year the currently approved milestones (indicators of research progress) Project 1: Genetic and environmental factors contributing to childhood obesity Year 1 2005: Complete enrollment and phenotyping of 300 families Repeat anthropometry and body composition measurements on 250 children Data entry and error checking Genotyping of first and second cohorts Preliminary quantitative trait loci (QTL) linkage analysis Year 2 2006: Repeat anthropometry and body composition measurements on 250 children Data entry and error checking Genotyping of third and fourth cohorts (780 individuals) QTL linkage analysis Publications Year 3 2007: Begin 1-y intervention trial for weight loss in overweight children Phenotyping including anthropometry and body composition of overweight children Data entry and error checking QTL linkage analysis Fine mapping and sequence positional candidate genes to identify single nucleotide polymorphisms (SNPs) Publications Year 4 2008: Continue 1-y intervention trial for weight loss in overweight children Phenotyping including anthropometry and body composition of overweight children Data entry and error checking QTL linkage analysis Fine mapping and sequence positional candidate genes to identify SNPs Publications Year 5 2009: Complete 1-y intervention trial for weight loss in overweight children Phenotyping including anthropometry and body composition of overweight children Data entry and error checking QTL linkage analysis Bayesian quantitative trait nucleotide (BQTN) analysis Publications Project 2: Biological diversity of human growth: body composition reference standards for children Year 1 2005: Recruitment for Cross-Sectional Pediatric Database Testing of different mathematical models as they relate to the Pediatric Reference Model Statistical analysis of percentile distribution models Extension of BodPod assay to include preschool ages and testing of PeaPod instrument for infants Year 2 2006: Continue recruitment of children for longitudinal databases Test prediction models with pediatric populations with known abnormal body composition Year 3 2007: Improvement of precision and accuracy of DXA software for use with infants and toddlers Continue longitudinal recruitment Identification of mathematical-based multi-parameter model that best describes changes in body composition during normal growth Year 4 2008: Continue longitudinal recruitment Preliminary test of the prediction accuracy of models using longitudinal database Test accuracy of forward prediction of Z-model with longitudinal database and disease populations Continued development of magnetic resonance technique for body water assay in preterm infants Year 5 2009: Continue longitudinal recruitment Adjust mathematical parameters to optimize models Update CNRC interactive website with latest Z-score prediction models Complete development of body composition analyzer for body fat, water, and protein analysis using a single scan at low dose Continued development of fat and protein analysis for infants and toddlers Project 3: Prevention of overweight in children Year 1 (2005) Submit request for OSQR Certification Year 2 (2006) Program design Program development Designing materials Participant recruitment and screening Continue participant recruitment and screening Complete baseline measurements on Phase 1 participants Randomize Phase 1 participants to intensive treatment or self-help treatment Begin treatment of Phase 1 participants Complete 6-month measurements on Phase 1 participants Begin analyses with Phase 1 6-month data Complete baseline measurements on Phase 2 participants Begin treatment of Phase 2 participants Continue treatment of Phase 2 participants Complete 12-month measurements of Phase 1 participants Begin analyses with Phase 1 12-month data Complete 6- and 12-month measurements of Phase 2 participants Complete study analyses Year 3 (2007) Pending based on certification of research project Project 4: Nutritional influences on innate immunity Year 1 2005: Define kinetics of macrophages in adipose depots and expression of mediators Determine the role of adhesion in RAW cell 3T3-L1 co-cultures Year 2 2006: Define kinetics of leukocytes, cytokines, adhesion molecules and chemokines in livers and adipose tissue of mice on high fat diet Determine the rate of reversal of pro-inflammatory conditions in liver and adipose tissue in mice following removal from high fat diet Determine the effect of probiotic microorganisms on liver injury in rats fed a high fat, high sucrose diet Determine the contribution of bone marrow precursors Year 3 2007: Determine which adipose sites respond to probiotic microorganisms Confirm that primary adipose macrophages modulate adipocyte function in vitro Determine the contribution of bone marrow precursors Determine mechanisms for apparently active reversal of pro-inflammatory conditions induced by high fat diet Year 4 2008: Determine the functional effects of macrophage inhibition Determine the synergy between dietary steatosis and endogenous endotoxin stimulus Year 5 2009: Determine the effects of differing lipids in the diet on macrophages in adipose tissue Determine the effects of differing lipids in the diet on synergistic liver injury Project 5: Effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents Year 1 2005: We anticipate that 20 subjects will have been studied. Year 2 2006: We anticipate an additional 20 subjects will have completed the 12 weeks exercise program and the pre-and post exercise metabolic studies. Preliminary data on potential metabolic effects of exercise in obese and lean subjects are available Year 3 2007: Study an additional 20 subjects, and continue data analyses Year 4 2008: Continue research studies and data analysis. Manuscript preparation. Year 5 2009: Complete all research studies, data analysis and manuscript preparation Project 6: Circadian clocks in adipose biology and obesity Year 1 2006: (RECEIVE OSQR CERTIFICATION) Isolate 24-hour adipose tissue samples Characterize expression of circadian clock genes in isolated tissue Isolate and prepare adipocytes for cell culture Initiate generation of transgenic animal Year 2 2007: Identify zeitgebers that regulate the adipocyte specific clocks Establish and maintain transgenic animal colony Perform microarray analysis in TG and WT animals Identify circadian clock-regulated genes Year 3 2008: Perform confirmatory Taqman assays Detailed characterization of adipocyte function and cellularity Initiate feeding studies of WT and TG animals Characterize weight gain and body composition changes in high- and low-fat fed animals Characterize gene expression patterns of clock mechanism genes Characterize gene expression patterns of clock regulated genes Year 4 2009: Identify timing of alterations in the circadian clock in relation to obesity onset Identify timing of alterations in clock regulated genes in relation to obesity onset 4a.List the single most significant research accomplishment during FY 2006. Project 1: Genetic and environmental factors contributing to childhood obesity Genetic and Environmental Factors influencing Childhood Obesity in the Hispanic Population The high prevalence of childhood obesity among American Hispanic children is a significant public health problem. Researchers at Children's Nutrition Research Center in Houston, TX, conducted a family-based study to identify both genetic and environmental factors affecting childhood obesity in 1030 children. Researchers used a multipoint genome scan to find and localize quantitative trait loci that influence variation in fat tissue content and obesity-related phenotypes in Hispanic children. This study has detected highly significant linkage signals on various chromosomes for insulin, waist to height ratio, glucose, and energy expenditure, respectively. The identified chromosomal regions harbor promising positional candidate genes, and we will resequence these genes in the parents to identify single nucleotide polymorphisms (SNPs) that contribute to the linkage signals on chromosomes; then the SNPs will be sequenced in the children to identify functional genetic variants. Identification of the significant genetic and environmental factors influencing childhood obesity in the Hispanic population will lead to more effective prevention and treatment of this growing epidemic. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] 4b.List other significant research accomplishment(s), if any. Project 1: Genetic and environmental factors contributing to childhood obesity Understanding Childhood Obesity in the Hispanic Population The high prevalence of childhood obesity among American Hispanic children is a significant public health problem. Scientists at the Children's Nutrition Research Center in Houston, TX, conducted a family-based study to identify both genetic and environmental factors affecting childhood obesity in 1030 children. The VIVA LA FAMILIA Study has enhanced our understanding of the epidemiology of childhood obesity in the Hispanic population. The intrinsic metabolic factors associated with childhood obesity and its metabolic complications have been described. We have identified birth weight, dietary patterns, physical activity, eating behavior as salient predictors of weight gain in this population. An enhanced understanding of the genetic and environmental factors influencing childhood obesity in the Hispanic population will lead to more effective prevention and treatment of the problem. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 2: Biological diversity of human growth: body composition reference standards for children Body composition assessment in early infancy Developing a better understanding of the effects on growth associated with different dietary choices early in life is needed. Scientists at the Children's Nutrition Research Center in Houston, TX, independently verified the accuracy of the infant-sized air-displacement plethysmograph (Pea Pod) for the measurement of body fatness. As a result, longitudinal monitoring of infants during the first six months of life can now be easily performed without a risk to the infant and its accuracy is second to none. This may assist in researchers' understanding the effects on growth associated with different dietary choices. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 2: Biological diversity of human growth: body composition reference standards for children Revised Z-score model for bone mineral mass in children Identifying a more accurate assessment of changes in bone mineralization during normal growth is needed to recognize children with an increased risk for fractures. Researchers at the Children's Nutrition Research Center in Houston, TX, completely reanalyzed bone mineral data maintained in the Children's Nutrition Research Center database for younger children. This effort addressed the problems related to the comparison of bone mineral values obtained with different dual energy X-ray absorption (DEXA) instruments and software versions. The on-line interactive Z-score prediction models available on the CNRC website have been updated for increased accuracy. As a result of analysis, a more accurate assessment of changes in bone mineralization during normal growth can now be obtained. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 3: Prevention of overweight in children Weight Loss in Mexican-American children The percentage of overweight children in the US is daunting, and a program to counteract this trend is needed. Children's Nutrition Research Center researchers conducted a study to evaluate an intensive school-based program designed to result in weight reduction for overweight Mexican-American children. Study results revealed that children in the Intensive Intervention condition significantly reduced their standardized BMI when compared to the children in the self-help condition. The change in standardized BMI was significantly different at both 3 and 6 months, with Intensive Intervention participants showing greater decreases in their standardized BMI. Overall, the results are promising, suggesting that an intensive school-based intervention may be an effective means for promoting weight loss in overweight Mexican-American children. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Adolescents Quality of Life Assessments Children's Nutrition Research Center study results showed very overweight adolescents demonstrated the lowest ratings for quality of life. When analyzing psychosocial and total quality of life scores, very overweight adolescents showed statistically lower functioning than all other groups, although for physical quality of life these adolescents were not statistically different from those classified as overweight. Overweight adolescents did not differ from those in any other weight category on physical quality of life scores. These results address the lack of information regarding quality of life of Mexican-American adolescents across weight classifications. Quality of life was assessed at baseline using an adolescent self-report, and measured height and weight were used to calculate body mass index (BMI) that was standardized for age and gender. Very overweight Mexican-American adolescents experience impairment in psychosocial and physical functioning when compared with normal weight peers. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 4: Nutritional influences on innate immunity Understanding Inflammatory Mediators in Adipose Tissue A greater understanding of the mechanisms by which obesity causes inflammatory mediators to be generated in adipose tissue is needed. Researchers at the Children's Nutrition Research Center found that the probiotic organisms in the commercial preparation VSL#3 given orally to rats significantly increased the plasma concentration of adiponectin, an anti-inflammatory mediator. In addition, VSL#3 administration resulted in probiotic colonization of the rat intestinal tract and significantly prevented liver injury caused by high fat diet and hindlimb unloading (a model that induces liver injury). Our researchers discovered that mice fed a diet rich in saturated fatty acids in addition to expressing pro-inflammatory markers also accumulated T cells. Subsets of these cells are of possible interest in the initiation or reversal of the pro-inflammatory state induced by a high fat diet. This is important as reseachers attempt to understand inflammatory mediators and their correlation with obesity. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Rapid changes in inflammatory mediators in adipose tissue. Obesity is a major health problem and there is growing evidence that high fat and high sucrose diets cause systemic inflammation that contributes to many of the complications of obesity such as arterial disease. Children's Nutrition Research Center researchers observed the development of inflammatory changes within 3 weeks in adipose tissue of mice fed a diet rich in saturated fatty acids. Our researchers discovered that the inflammatory changes reversed significantly faster than loss of fat mass when the mice were returned to a low fat diet. The specific markers of inflammation included chemokines such as RANTES (previously unknown in adipose tissue), cytokines such as IL-6 (occurring before TNF), and macrophages (a subset of activated macrophages previously not described by other investigators). These results may relate to the observations in humans of significant improvements in systemic inflammatory markers with relatively little weight loss. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 5: Effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents Preliminary data suggest health impact, without weight loss intent Over the past 30 years the prevalence of overweight children in the USA has increased 3-fold to reach 15% in 2000, with an even higher incidence in Hispanic and African American adolescents. Scientists at the Children's Nutrition Research Center in Houston, TX, have conducted a 12-week exercise program without the intent of losing weight. Preliminary data demonstrate that. 1)all subjects increased their fitness following the 12-week exercise program;. 2)Although total fat mass did not decrease, visceral fat (one of the most metabolically active fat depots) decreased in all subjects;. 3)Liver fat decreased in some subjects, particularly those who had the highest values; and. 4)Fasting insulin decreased, indicating improved insulin sensitivity. Researchers conclude that a 12-week exercise program without intent to weight loss improved insulin sensitivity by reducing visceral fat and increasing fitness. Additionally, it might alleviate fat dispositions on the liver, which is a serious obesity complication. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] Project 6: Circadian clocks in adipose biology and obesity Developing a Functional Circadian Clock within Adipose Prior to the initiation of our experiments, the presence of a functional circadian clock within adipose had not been fully demonstrated. Researchers at the Children's Nutrition Research Center, Houston, TX, have established the presence of a fully functional clock within adipose, and several groups have also reported similar results. This was accomplished through isolating adipose tissue over the course of 24 hours while measuring the gene expression in circadian clock components. Our results suggest that lipid metabolism in adipose and other tissues may be directly regulated by the circadian clock. The establishment of the adipose-specific circadian clock provides the basis for future experiments designed to assess the role of this clock in metabolism and obesity. [NP107-Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease] 4c.List significant activities that support special target populations. None. 5.Describe the major accomplishments to date and their predicted or actual impact. This research contributes to the goals of NP107, Human Nutrition, specifically "Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease". It address ARS Strategic Plan Goal #4 Improve the Nation's Health, specifically objective 4.1: Promote Healthier Individual Food Choices and Lifestyles and Prevent Obesity; Improve Human Health by Better Understanding the Nutrient Requirements of Individuals and the Nutritional Value of Foods; Determine Food Consumption Patterns of Americans. Project 1: Genetic and environmental factors contributing to childhood obesity Our research project has made significant progress in understanding the environmental and genetic factors affecting childhood obesity in the Hispanic population in the U.S. Our major goal was to identify and phenotype/characterize 300 overweight Hispanic probands (ages 4-19 y) and his/her biological parents and siblings with respect to adiposity, the regulation of food intake, energy expenditure, and energy partitioning. We have completed the phenotyping and genotyping of a total of 1030 children and 631 parents from 319 Hispanic families in our VIVA LA FAMILIA study. Genome scan using variance component linkage analysis has been performed on our initial set of traits. We have found highly significant linkage signals on a number of chromosomes for obesity and its mediators of diet and physical activity, and its comorbidities. Project 2: Biological diversity of human growth: body composition reference standards for children Accurate assessment of changes in body composition of the individual child, especially that of body fatness, is needed in order to advance our understanding of the influences of variations in the nutrient and activity patterns associated with normal growth. Deviations in these patterns may be early indicators of increased risks for the development of childhood obesity. The technologically advanced instruments available at the CNRC, coupled with the on-going longitudinal studies in selected pediatric populations, are needed to accomplish this task. In addition, the continued evaluation and cross-calibration of smaller portable devices will help other scientists make better choices where assessment of body fatness is needed in large epidemiology and/or intervention studies. Project 3: Prevention of overweight in children Analyses have demonstrated that participants who participated in the intensive intervention are not as heavy as the children assigned to the self-help program. Additionally, children in the intensive intervention have improved their quality of life, cholesterol levels, and diet quality. Preliminary analyses have shown that these results are being maintained after 12 months. We will present these results to scientists, clinicians, and public health authorities at an international meeting of behavioral change researchers in November 2006 and in the form of several manuscripts that have already been submitted for publication. These customers will gain valuable information regarding how to solve the problem of recruiting and retaining Mexican-American children and their families in research. In addition, they will benefit from strategies and techniques that will be beneficial in addressing the problem of childhood obesity in Mexican-Americans. Project 4: Nutritional influences on innate immunity We have found that short-term feeding of high fat diets results in pro-inflammatory conditions in the liver and adipose tissue of mice and rats. In both species, males are affected more profoundly than females by the short-term diets of 3-5 week duration. Long-term high fat diets result in pro-inflammatory conditions similar to males in the adipose and liver. We have identified previously unknown inflammatory markers. These include chemokines, RANTES and eotaxin, a subpopulation of activated macrophages expressing CD11c, lymphocytes of the gamma delta T cell subclass. We found that liver injury is increased by hindlimb unloading, a model that induces portal endotoxemia, and that the endotoxin coming from the GI tract augments the liver injury induced by the high fat diet. We found that probiotic bacteria can prevent this liver injury. We found that the pro-inflammatory condition in mouse adipose tissue is rapidly reversed when animals are returned to a low fat diet and this reversal is significantly more rapid than the reduction in fat mass. These data support the use of mouse and rat models of obesity as models for analysis of mechanisms related to human obesity and the metabolic syndrome. The project overall has addressed the problem of dietary obesity and its ability to produce systemic inflammation and liver injury. In addition to the accomplishments listed above, our researchers have found that endotoxin released from the gut contributes to the liver injury, and we have defined the contributions of adhesion molecules to the emigration of leukocytes. We are defining the cascade of events that leads to systemic inflammation and liver injury. These are major complex processes and have not been solved. We have defined characteristics of macrophages in fat. We have defined the major inflammatory cell type in the obesity liver. We have shown that stress increases liver damage in animals on high fat diet. We have found sex differences in the liver and adipose tissue response to high fat diet. In addition, we have extended the models to demonstrate beneficial effects of probiotic bacteria and the rapid reversal of pro-inflammatory conditions was modest reductions is adipose tissue mass. Project 5: Effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents Scientists at the Children's Nutrition Research Center in Houston, TX, have showed that the exercise program designed for this study is well accepted by both obese and lean subjects. The subjects have complied very well to the program, both at the Wellness Center and at home, resulting in increased fitness in all subjects (average 17% increase in VO2 max). Researchers have also refined the MRI measures of intramyocellular, visceral, and hepatic fat. Preliminary data showed that obese subjects had about twice as much intramyocellular, hepatic, and visceral fat as compared to their lean counterparts; in the obese subjects, the most significant fat reduction resulting from the exercise program occurred in visceral and liver fat; also insulin concentrations were about twice as high in the obese as compared to the lean subjects and decreased significantly in the obese with no change in the lean adolescents. Since we are in the early stage of this project, the primary customers are the participating adolescents. However, the data were presented at a national meeting that is attended by a large number of pediatricians. The subjects involved in these studies live a sedentary life style. Virtually all of them have indicated that they have enjoyed participating in the program and that they have felt good about themselves for being able to fulfill the program. They have also indicated their intention to continue exercising after completion of the exercise. These results are important since these adolescents, particularly the obese, most often avoid participating in physical education, kinesiology, or other athletic activities. Project 6: Circadian clocks in adipose biology and obesity We have characterized the basic components of the circadian mechanism within the adipose cell and have demonstrated this mechanism to be fully functional within this cell type. We are in the process of creating an adipose-specific dominant negative clock transgenic mouse. Its successful creation will provide an important model for elucidating the role of the circadian clock within adipose in global metabolism and the development of adiposity. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Project 2: Biological diversity of human growth: body composition reference standards for children The Z-score prediction model for bone assessment in children is available on-line in an interactive mode as part of the Children's Nutrition Research Center's web-site www.kidsnutrition.org. More than 300 national and international users have registered for access to this feature. Project 3: Prevention of overweight in children This program has been given to the school in which it was conducted for use in adapting their health class, and the research has been disseminated in the form of conference presentations and scientific publications. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Project 1: Genetic and environmental factors contributing to childhood obesity NF Butte. "Early metabolic syndrome," North American Association for Studies on Obesity Annual Meeting, Vancouver, Canada, October 2005. N.F. Butte. "Dietary reference intakes for energy: infants and children," American Dietetic Association, St. Louis, MO, October 2005. N.F. Butte. "Genetic and environmental factors contributing to childhood obesity in the Hispanic population," 1st Regional Congress on Childhood Obesity, Monterrey, Mexico, November 2005. N.F. Butte. "Environmental and genetic contribution to the development of obesity in Hispanic children and adolescents- VIVA LA FAMILIA Study," 45th Annual Meeting Mexican Society of Nutrition and Endocrinology, Merida, Yucatan, Mexico, November-December 2005. N.F. Butte. "Interventions for the prevention of childhood obesity," UN regional expert meeting to elaborate new regional project proposal on childhood obesity, Panama City, Panama, May 2006. N.F. Butte. "Genetics of childhood obesity: VIVA LA FAMILIA Study," American Diabetes Association, Washington DC, June 2006. Project 2: Biological diversity of human growth: body composition reference standards for children K.J. Ellis. Invited oral presentation entitled "Body composition assessment techniques" at the 46th Annual Meeting of the European Society for Pediatric Research (Siena, Italy, September 2005) Project 3: Prevention of overweight in children Johnston, C.A., Tyler, C., Fullerton, G., Carvalho, S.M., El-Mubasher, A.A., Volding, D., Poston, W.C., Haddock, C.K. Reeves, R., Foreyt, J.P. (November 2006). School-based Weight Management: Outcomes with Mexican American Adolescents. Presented at the International Congress of Behavioral Medicine's Ninth Meeting, Bangkok, Thailand. Fullerton, G., Tyler, C., Johnston, C.A., El-Mubasher, A.A., Carvalho, S.M., Volding, D.C., Reeves, R., Foreyt, J.P. (November 2006). Quality of Life in Overweight Mexican American Children. Presented at the International Congress of Behavioral Medicine's Ninth Meeting, Bangkok, Thailand. Tyler, C., Fullerton, G., Johnston, C.A., Ortega, A. (October 2005). Weight Classification and Quality of Life in Mexican American Children. Presented at NAASO's 2005 Annual Meeting, Vancouver, BC. Johnston, C.A., Tyler, C., Foreyt, J.P., Reeves, R., Poston, W.S.C., Haddock, C.K., Ortega, A., Fullerton, G. (October 2005). Evaluation of an Extended Day Weight Loss Program for Mexican American Children. Presented at NAASO's 2005 Annual Meeting, Vancouver, BC. Ortega, A., Johnston, C.A., Tyler, C., Carvalho, S.M. (October 2005). Acculturation and Weight Status in Mexican American Children. Presented at NAASO's 2005 Annual Meeting, Vancouver, BC. Johnston, C.A., Tyler, C., Miller, F., Dadjoo, N., McFarlin, B.K. (October 2005). Metabolic Syndrome Risk Across Weight Status in Mexican American Children. Presented at NAASO's 2005 Annual Meeting, Vancouver, BC. Project 5: Effects of dietary macronutrient distribution and exercise on glucose metabolism in obese adolescents A. Sunehag. The data were presented at the Annual meeting for the Society of Pediatric Research in San Francisco, CA, April 30, 2006. Review Publications Zakeri, I., Puyau, M.R., Adolph, A.L., Vohra, F.A., Butte, N.F. 2006. Normalization of energy expenditure data for differences in body mass or composition in children and adolescents. Journal of Nutrition. 136(5):1371-1376. Butte, N.F., Mehta, N., Ellis, K.J., Cole, S., Bastarrachea, R., Comuzzie, A. 2004. Predictors of the metabolic syndrome in overweight Hispanic children. Obesity Research. S12(10):A144. Bastarrachea, R., Tejero, M., Cai, G., Cole, S., Mehta, N., Ellis, K.J., Butte, N.F., Comuzzie, A. 2004. Pleiotropy effects on endocrine traits and body composition in children. Obesity Research. S12(10):A194. Cai, G., Tejero, M., Cole, S., Mehta, N.R., Butte, N.F., Comuzzie, A. 2004. Evidence of pleiotropy between obesity and other phenotypes related to the metabolic syndrome in children. Obesity Research. S12(10):A194. Butte, N.F., Comuzzie, A.G., Cole, S.A., Mehta, N.R., Tejero, M., Bastarrachea, R., Smith, O.E. 2005. Quantitative genetic analysis of the metabolic syndrome in hispanic children. Pediatric Research. 58(6):1243-1248. Butte, N.F. 2005. Energy requirements of infants. Public Health Nutrition. 8(7A):953-967. Butte, N.F., King, J.C. 2005. Energy requirements during pregnancy and lactation. Public Health Nutrition. 8(7A):1010-1027. Sunehag, A.L., Wang, Z.J., Green, T.F., Bier, D.M., Haymond, M.W. 2006. A 12 weeks exercise program resulted in reduced visceral fat and fasting insulin but not total and intramyocellular fat in Hispanic obese adolescents [absract]. 2006 Pediatric Academic Societies Meeting. Paper No. 753346. Foreyt, J.P. 2005. Need for lifestyle intervention: how to begin. American Journal of Cardiology. 96(4A):11E-14E. Pinkston, M.M., Poston, W.S.C., Reeves, R.S., Haddock, C.K., Taylor, J.E., Foreyt, J.P. 2006. Does metabolic syndrome mitigate weight loss in overweight Mexican American women treated for 1-year with orlistat and lifestyle modification? Eating and Weight Disorders. 11(1):e35-e41. Rivera, C.A., Abrams, S.H., Tcharmtchi, M.H., Allman, M., Ziba, T.T., Finegold, M.J., Smith, C.W. 2005. Feeding a corn oil/sucrose-enriched diet enhances steatohepatitis in sedentary rats. American Journal of Physiology - Gastrointestinal and Liver Physiology. 290:G386-G393. Falanga, A., Marchetti, M., Barbui, T., Smith, C.W. 2005. Pathogenesis of thrombosis in essential thrombocythemia and polycythemia vera: The role of neutrophils. Seminars in Hematology. 42:239-247. Xiong, C., Hixson, P.M., Mendoza, L.H., Smith, C.W. 2005. Cloning and expression of rabbit interleukin-15. Veterinary Immunology and Immunopathology. 107:131-141. Blair, C.K., Folsom, A.R., Knopman, D.S., Bray, M.S., Mosley, T.H., Boerwinkle, E. 2005. APOE genotype and cognitive decline in a middle-aged cohort. Neurology. 64:268-276. Bray, M.S., Ellis, K.J., Bush, J.A., Turpin, J., Callie, M., Miller, F., Jackson, A. 2005. Race and ethnic effect of estimating DXA percent fat from BMI: The Tiger Study [abstract]. Medical Science Sports Exercise. 37(5 Supl):S303. Terre, L., Poston, W.S.C., Foreyt, J., St. Joer, S.T., Horrigan, K.L. 2004. Does family of origin functioning predict adult somatic complaints? Psychology and Health. 19(4):507-514. Conard, M.W., Poston, W.S.C., Foreyt, J.P. 2005. Managing obesity in primary care. In: O'Donohue, W.T., Byrd, M.R., Cumming, N.A., Henderson, D.A., editors. Behavioral Integrative Care: Treatments That Work in the Primary Care Setting. New York, NY: Brunner-Routledge. p. 253-271. Terre, L., Poston, W.S.C., Foreyt, J.P. 2006. Eating disorders. In: Mash, E., Barkley, R., editors. Treatment of Childhood Disorders. 3rd edition. New York, NY: Guilford Press. p. 778-829. Morris, A.P., Tawil, A., Berkova, Z., Wible, L., Smith, C.W., Cunningham, S.A. 2006. Junctional adhesion molecules (JAMs) are differentially expressed in fibroblasts and co-localize with ZO-1 to adherens-like junctions. Cell Communication and Adhesion. 13:233-247. Sunehag, A., Haymond, M.W. 2005. Persistant hyperinsulinemic hypoglycemia of infancy. UUpToDate Online Journal [serial online]. Available: http://uptodateonline.com. Sunehag, A., Haymond, M.W. 2005. Etiology of hypoglycemia in infants and children. UpToDate Online Journal [serial online]. Available: http://uptodateonline.com. Terre, L., Poston, W.S.C., Foreyt, J.P. 2004. Overview and the future of obesity treatment. In: Goldstein, D.J., editor. The Management of Eating Disorders and Obesity. 2nd edition. Totowa, NJ: Humana Press. p. 161-179. Sunehag, A.L., Haymond, M.W. 2005. Approach to hypoglycemia in infants and children. UpToDate Online Journal [serial online]. Available: http://uptodateonline.com. Wolfarth, B., Bray, M.S., Hagberg, J.M., Perusse, L., Rauramaa, R., Rivera, M.A., Roth, S.M., Rankinen, T., Bouchard, C. 2005. The human gene map for performance and health-related fitness phenotypes: the 2004 update. Medicine and Science in Sports and Exercise. 37(6):881-903. Cobain, M.R., Foreyt, J.P. 2005. Designing "lifestyle interventions" with the brain in mind. Neurobiology of Aging. 26S:S85-S87. Lafontant, P.J., Burns, A.R., Donnachie, E., Haudek, S.B., Smith, C.W., Entman, M.L. 2006. Oncostatin M differentially regulates CXC chemokines in mouse cardiac fibroblasts. American Journal of Physiology - Cell Physiology. 291:C18-C26. Poston, W.S.C., Haddock, C.K., Pinkston, M.M., Pace, P., Karakoc, N.D., Reeves, R.S., Foreyt, J.P. 2005. Weight loss with meal replacement and meal replacement plus snacks: a randomized trial. International Journal of Obesity. 29(9):1107-1114. Olutoye, O.O., Zhu, X., Cass, D.L., Smith, C.W. 2005. Neutrophil recruitment by fetal procine endothelial cells: Implications in scarless fetal wound healing. Pediatric Research. 58:1290-1294. Perri, M.G., Foreyt, J.P. 2003. Preventing weight regain after weight loss. In: Bray, G., Bouchard, C., editors. Handbook of Obesity. New York: Marcel Dekker. p. 185-199. Poston, W.S.C., Foreyt, J.P. 2004. Sibutramine and the management of obesity. Expert Opinion in Pharmacotherapy. 5(3):633-642. Ioannidis, J.P., Gwinn, M., Little, J., Higgins, J.P.T., Bernstein, J.L., Boffeta, P., Bondy, M., Bray, M.S., Brenchley, P.E., Buffler, P.A. 2006. A road map for efficient and reliable human genome epidemiology. Nature Genetics. 38(1):3-5. Sturgeon, J.D., Folsom, A.R., Bray, M.S., Boerwinkle, E., Ballantyne, C.M. 2005. Apolipoprotein E genotype and incident ischemic stroke: The Atherosclerosis Risk in Community Study. Stroke. 36:2484-2486. Ellis, K.J. 2003. Assessment of body composition in children with Prader-Willi Syndrome or simple obesity. In: Eiholzer, U., I'Allemand, D., Zipf, W.B., editors. Prader-Willi Syndrome as a model for obesity. New York:Karger. p. 49-60. Li, Z., Burns, A.R., Smith, C.W. 2006. Two waves of neutrophil emigration in response to corneal epithelial abrasion: Distinct adhesion molecule requirements. Investigative Opthalmology and Visual Science. 47:1947-1955. Pritchett, A.M., Foreyt, J.P., Mann, D.L. 2005. Treatment of the metabolic syndrome: the impact of lifestyle modification. Current Atherosclerosis Reports. 7(2):95-102. Dhillon, S.S., Mahadevan, K., Bandi, V., Zheng, Z., Smith, C.W., Rumbaut, R.E. 2005. Neutrophils, nitric oxide, and microvascular permeability in severe sepsis. Chest. 128:1706-1712.