2005 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? What does it matter? With the promotion of the "Food Guide Pyramid" by the Dietary Guidelines for Americans, more plant-based foods are being consumed. Vegetables and fruits contain substances, phytochemicals, that alter certain body functions and ultimately are responsible for improved health. We are studying phytochemicals in many foods: fruits, rice, and soybeans. There are many general types of phytochemicals, such as isoflavones, also known as "phytoestrogens". Isoflavones are particularly concentrated in soybeans and can have many of the same actions as the major female hormones in women, the estrogens. In countries where people regularly eat large amounts of soy products, several health advantages have been recognized, including lower risk of several cancers and other chronic diseases, and isoflavones appear to be partially responsible. Approximately one million US infants are fed infant formula containing soy protein each year and the phytoestrogen content in their blood is extremely high. Children usually do not have high blood estrogen levels until puberty. Virtually nothing is known about the actions of these substances in children, nor on the long-term health consequences of this early exposure. Some suggest that early soy intake will improve health, and others suggest it is dangerous to infants and could produce health problems later in life. Some countries recommend that no soyfoods be consumed before 6 to 36 months. Since this affects nearly 25% of America's youngest people, it is very important to understand the long-term health consequences of early consumption of these phytochemicals. We hypothesize that there will be both positive and negative effects of early consumption of high levels of soy foods, and it is essential to define these. For example, we have evidence suggesting that diets rich in soy products will reduce the risk of cancers (such as breast and colon cancer) later in life. Similarly, we suspect that there may be some as yet unrecognized effects related to hormonally sensitive systems that regulate metabolism that result in changes in drug efficacy. We have developed a team of basic and clinical investigators to conduct laboratory, animal, and clinical studies to test these hypotheses. We anticipate that findings from this research will stimulate U.S. food processors to develop and market several new food products containing soy, rice, fruits, and milk proteins to maximize the health in children now, and in their later years. The questions being addressed in this project are very important and consistent with the National Program - 107 goals. This research directly supports ARS Strategic Plan Goal #4, Improve the Nation's Nutrition and Health. It is extremely important to determine the positive and negative health consequences of various dietary factors (phytochemicals), such as soy phytoestrogens in infant formula, especially as it relates to safety and disease prevention. It is essential that we understand which phytochemicals have health consequences, how much and how often they should be consumed and at what age they should be eaten. It is important to know whether it is possible to prevent cancer and cardiovascular diseases and under what circumstances. This would improve the health and well-being of Americans and reduce health care costs. It is important to know if soy formula is safe for children. 2.List the milestones (indicators of progress) from your Project Plan. Research funded by this CRIS concentrates on one theme, the long-term health effects of non-nutrient factors in food, particularly as related to human development and health. We are interested in how these dietary factors affect early development of children and whether early exposure to these factors will reduce the risk of chronic disease that typically occur later in life. There are three general areas of research directed towards normal growth, development, and disease prevention:. 1)cancer prevention (Project 1);. 2)prevention of cardiovascular disease (Project 2);. 3)metabolism, endocrinology, growth, and body composition (Project 3). These projects are aimed at studies of those foods consumed by infants or children, such as soy infant formula, milk proteins, fruits, and other grains such as rice. Project 1: Effects of Dietary Factors on Cancer Prevention. This research will elucidate the effects of early consumption of dietary factors on normal development and function of the mammary glands and gastrointestinal tract. It will further determine the protective effects of this early exposure to dietary factors on mammary and colon cancer. Importantly, this project will determine the molecular mechanism(s) underlying the developmental and protective effects of dietary factors on adult risk of mammary and colon cancer using rat models of chemical carcinogenesis. Because a high percentage of infants and children consume large amounts of soy infant formula and/or milk, our initial studies will focus on lifetime consumption and exposure at specific developmental windows to the three proteins consumed by infants (soy protein isolate, whey protein, and casein) on adult tumor incidence, tumor latency, and tumor multiplicity in rats exposed to carcinogens. We will also conduct studies into the effects of fruits commonly consumed by children. These studies will constitute a comprehensive evaluation of the biological and molecular mechanisms underlying the interaction of diet and developmental status on adult breast and colon and the cancer risks on these organs. Year 1 (FY 2004): Office of Scientific Quality Research (OSQR) peer review of the project, consisting of this research project, is completed and certified. Year 2 (FY2005): Determine the effects of soy and whey on development and function of mammary gland and gastrointestinal (GI) tract. Initiate studies on effects of fruits on growth and development of rats through the lifecycle. Demonstrate that soy and/or whey confer protection against chemically induced mammary and colon tumorigenesis by mechanisms independent of pro-carcinogen activation. Determine if induction of the apoptotic status of mammary epithelial cells in young adults (PND50 rats) by soy and whey is one likely mechanism for mammary tumor protection. Identify one or more molecular pathways altered by whey and/or soy that are associated with the observed increased epithelial cell apoptosis. Determine if the rat aberrant crypt foci (ACF) model provides results similar to those from previous studies of rat colon tumors in response to dietary factors. Initiate studies on colon cancer preventive effects of fruits in rats. We anticipate several manuscripts and/or abstracts will be submitted. Year 3 (FY2006): Demonstrate that early exposure only (i.e., in utero, perinatal, peripubertal only) to dietary soy mimics the mammary and/or colon cancer protective effects conferred by lifetime exposure. Determine if dietary genistein exposure in utero confers protection against mammary tumorigenesis. Determine if lifetime dietary soy intake predisposes young adults to adverse reproductive phenotypes. Evaluate the role of up-regulated expression of the tumor suppressor p53 in the tumor protective effects of whey proteins. Determine the effects of soy and whey proteins on normal gut development. Determine the potential protective effects of blueberries against colon cancer. Begin studies on tumor immunosurveillance underlying cancer prevention by soy and whey. Begin experiments towards developing a pig model of gastrointestinal development and for studies into the gut-associated lymphoid tissue (GALT) system. Use the pig model to initiate studies on the effects of diets on development and modulation of the GALT. We anticipate several manuscripts and/or abstracts will be submitted. Year 4 (FY2007): Utilize microarray technology to determine the gene expression profiles of mammary epithelial cells and cells from the GI tract from young adult rats lifetime exposed to dietary soy and whey proteins, relative to casein-fed rats. Identify one or more signaling pathways that are altered by these diets from analysis of the microarray data, and which will not otherwise be identified by the candidate gene approach. Begin to identify serum markers for tumor protection in young adult rats fed soy, whey, and/or fruit diets. Begin studies on fetal gut growth and differentiation resulting from soy diets and/or diets made with soy isoflavones. Determine if soy and/or whey protects against colon cancer by favoring apoptosis of cells with DNA damage after administration of carcinogen. Begin to address the functional regulation of Natural Killer (NK) cells and Dendritic Cells (DC) by dietary factors as possible molecular mechanisms underlying the cancer preventive effects of soy and other diets. We anticipate submitting several manuscripts and/or abstracts on the results of these studies. Year 5 (FY2008): Evaluate whether dietary exposure to soy and/or whey, relative to casein-fed rats at neonatal or peri-pubertal stage only, confer protection from chemically-induced mammary tumorigenesis. Determine whether one of the identified molecular pathways altered by lifetime exposure to dietary soy or whey (see year. 4)is similarly utilized for this cancer protection. Examine whether soy and/or whey regulate the nucleotide excision repair pathway as one potential molecular mechanism for tumor protection. Examine the protective actions of dietary soy and/or whey proteins on carcinogen metabolism and DNA damage in the rat colon model. Determine how dietary soy and/or whey proteins regulate intestinal gene expression. Continue our studies on GALT development and maturation using a swine model. We anticipate submitting several manuscripts and/or abstracts on the results of these studies. Project 2: Effects of Dietary Factors on Prevention of Atherosclerosis. Cardiovascular disease (CVD) is the number one killer of Americans. It is well known that certain CVDs, such as atherosclerosis, start very early in life, even before birth. This research will elucidate the effects of early consumption of dietary factors on development of CVD. We will concentrate on soy, rice, and fruits, because epidemiological data suggests that consumption of these foods prevents CVD. The project will determine the effects of this early exposure to dietary factors on experimentally induced atherosclerosis in animal models. Thus, we will study the effects of these foods in animals during pregnancy (fetal exposure), in neonates (perinatal) and other periods of early life (i.e., the peripubertal period). Our preliminary evidence suggests that phytochemicals bound to soy protein and rice protein, as well as peptides in whey protein hydrolysate and in soy protein isolates, prevent experimentally induced atherosclerosis. The main point of this project is to determine if early exposure to these and other potentially preventive phytochemicals in foods consumed by pregnant women or children can prevent cardiovascular diseases. Importantly, this project will determine the molecular mechanism(s) underlying the protective effects of dietary factors on risks associated with cardiovascular diseases using state-of-the-art animal models and molecular techniques. Year 1 (FY 2004): Office of Scientific Quality Research (OSQR) peer review of the project, consisting of this research project, is completed and certified. Year 2 (2005): Conduct in vivo experiments to determine if soy and whey diets will prevent the early onset of atherogenesis using atherosclerosis prone apolipoprotein E (apoE) knockout mouse (apoE-/-). Determine the effect of soy-based diets on lipid profiles and other immunological parameters. Begin analyses of aorta samples to determine experimental effects. Determine whether soy phytochemicals and/or soy proteins mediate protective effects of soy-based diet. Determine the effect of soy phytochemical on monocyte adhesion to endothelial cell adhesion molecule. Obtain sufficient data to present an abstract at scientific meetings. Year 3 (2006): Complete sample analyses from in vivo experiments in Year 1 on athero-protective effects of soy-based and whey-based diets. Initiate the in vivo experiments on the athero-protective effects of rice protein isolate. Start analyses of aorta samples from in vivo rice protein isolate study if there are protective effects. Determine the effect of rice protein isolate on lipid profiles and other immunological parameters. Evaluate the regulation of genes expressed in endothelial cells involved in atherosclerosis by soy-based diets. Determine modulation of macrophage specific genes by soy-based diets. Establish maternal hypercholesterolemia-induced atherogenesis in F1 generation using apoE-/- mouse model. Initiate studies on hyperhomocysteinemia-induced atherogenesis using apoE-/- mouse model. I nitiate in vivo studies on the potential protective effects of fruit-containing diets against atherogenesis using apoE-/- mouse model. Prepare and submit a manuscript on soy preventive effects on atherosclerosis. Obtain sufficient data to present an abstract at a national meetings. We anticipate submitting multiple manuscripts and/or abstracts on the findings from these studies. Year 4 (2007): Utilize gene microarray technology to determine the gene expression profiles of aortic endothelial cells and macrophage from apoE-/- mouse fed soy-based diets. Determine whether the soy-mediated athero-protection is mediated by regulation of endothelial cell specific cell adhesion molecules. Demonstrate the effects of soy-based diets on regulation and functions of macrophages isolated from apoE-/- mouse exposed to soy-based diets. Determine the role of soy-based diets on the induction of antibody against oxidized-LDL. Evaluate the anti-inflammatory property of soy by analyzing the expression of pro- and anti-inflammatory cytokines/chemokines. Begin to address whether soy-based diets prevent hyperhomocysteinemia-induced atherosclerosis. Obtain sufficient data to present the findings from these studies at a national meeting. We anticipate multiple manuscripts/abstracts will be submitted. Year 5 (2008): Demonstrate whether rice protein isolate has anti-inflammatory properties, one of the possible mechanisms for its protective effects. Determine if lifetime exposure of soy/rice confers the protective effect in maternal hypercholesterolemia-induced atherosclerosis in the F1 generation. Determine if soy photochemical and/or soy proteins mediate the protective effects. Demonstrate the effects of early exposure (in utero) of soy or rice protein isolate prevents atherosclerosis in maternal hypercholesterolemia model. Determine if soy exposure during different developmental windows (perinatal, peribubertal only) confers athero-protective effects in maternal hypercholesterolemia model. Present the findings from these studies at a national meetings. We anticipate submitting multiple manuscripts on the results of these studies. Project 3. Effects of Dietary Factors on Metabolism, Endocrinology, Growth and Body Composition. Much available data suggests that exposure to exogenous compounds, such as the phytochemicals found in fruits, vegetables, and grains, can alter metabolism and the levels and actions of hormones, all of which can: A) affect growth, development and body composition [including fat deposition and obesity prevention]; and B) can influence the risk of chronic diseases that occur both during development and later in the aging process. This research project studies the foods fed to newborns and especially addresses the three major proteins consumed by infants (casein, whey, and soy proteins). In addition, rice protein is studied because there is recent evidence of potential health benefits and it may become a substitute for soy protein in infant formulas and baby foods. With the increased emphasis on greater levels of fruit intake for developing children, we will also focus on health benefits of fruits and phytochemicals in fruits. The focus of this project is on the non-nutrient dietary factors in foods (phytochemical composition, bioavailability, organ distribution, and metabolism) and the biochemical effects of these diets. The research approach focuses on: the molecular mechanisms underlying dietary modulation of enzymes in the cytochrome P450 gene families which are involved in cholesterol metabolism and the metabolism of pediatric drugs and other xenobiotics; and the effects and mechanisms of dietary factors on regulation of healthy body composition and obesity prevention. Year 1 (FY 2004): Office of Scientific Quality Research (OSQR) peer review of the project, consisting of this research project, is completed and certified. Year 2 (FY 2005): Determine soy effects on CYP3A expression and inducibility in young rats. Determine the effects of soy protein isolate and isoflavones on cholesterol metabolism and transport. Begin studies on health effects of early consumption of rice protein isolate. Determine the content of phytochemicals in soy protein isolate, fruits, and rice protein isolate. Determine the isoflavone content of serum, urine, and tissues following consumption of soy protein isolate. Collect blood and urine samples from infants fed soy infant formula. Conduct body composition studies on infants fed breast milk or formula. Track body weight, body length, and head circumference of infants to identify children at risk for obesity. Conduct body composition and metabolism studies on rats fed high-fat diets and diets with differing phytochemical composition. Begin investigations into establishing a pig model to study the health effects of infant formula. Determine changes in body composition resulting from high-fat diets in rats. Begin studies on the development of hepatic steatosis and NASH. Investigate the effects of alterations in the type of dietary fat (saturated, monounsaturated, or polyunsaturated) in the development of NASH and metabolic syndrome. Present data on: soy effects on cholesterol metabolism and transport; interspecies isoflavone metabolism; organ distribution of isoflavones; and rice effects on gene regulation at national meetings, and publish at least one paper on soy effects on CYP3A. Year 3 (FY 2006): Continue studies on biochemical and body composition effects of soy protein isolate and rice protein isolates. Continue studies to determine the mechanisms by which soy alters CYP1A and reduces the Aryl Hydrocarbon receptor. Analyze urine from infants fed soy infant formula for isoflavone, cortisol, and 6 Beta-hydroxycortisol. Begin analyses of isoflavones in pooled blood samples from infants fed soy formula. Continue studies on synergistic induction of CYP3A. Investigate the effects of antioxidants on the development of NASH via high-fat diets. Examine the roles of PPAR-Y and PPAR-A in alleviating NASH induced by high-fat diets. Examine if in utero exposure to maternal obesity programs higher birth weight and weight gain in rodents (rats and mice). Determine whether in utero exposure to maternal obesity alters neonatal leptin surge and programs leptin resistance in the offspring. Attempt to transfer and standardize the neonatal total enteral nutrition rat model for use in infant formula and obesity studies. Continue the development of the animal model for infant formula studies and once standardized, begin studies. Submit papers on effects of soy on CYP3A and cholesterol transport. We anticipate submitting several manuscripts and/or abstracts on the results of these studies. Year 4 (FY 2007): Continue developmental studies in piglets on the effects of dietary factors in soy, rice, whey, and fruits on CYP3A and metabolism of endogenous substrates and xenobiotics (such as pediatric medications). Determine the effects of the soy components (genistein and daidzein) on steady-state AhR expression and CYP1A1 induction. Examine effects of anti-inflammatory agents on the development of NASH. Demonstrate if greater early weight gain (due to overnutrition or catch-up growth) alters the body-weight set point via leptin resistance in rodent offspring. Examine the interventional potential of dietary factors in mitigating the higher body-weight set-point. Examine if greater adiposity in rodents is due to in utero or early-life programming is associated with increased stem cell commitment and/or differentiation in the adipose tissue. We anticipate submitting several manuscripts and/or abstracts on the results of these studies. Year 5 (FY 2008): Determine the effect of neonatal nutritional exposure to dietary factors (to mimic exposure to soy and milk proteins during lactation) and its impact on future weight gain and metabolic set-point. Employ microarrays as a hypotheses generator to determine gene expression profiles and identify endocrine, cellular, and molecular regulators mediating in utero and neonatal programming of dietary factors in the adipose tissue. Using a candidate gene approach, examine specific endocrine and cellular signaling changes in the liver, muscle, and adipose tissue of adult animals. Determine the effects of soy consumption on activated aryl hydrocarbon receptor-estrogen receptor crosstalk. We anticipate submitting several manuscripts and/or abstracts on the results of these studies. 4a.What was the single most significant accomplishment this past year? MECHANISMS BY WHICH SOY PROTEIN REDUCES CHOLESTEROL Investigators at the Arkansas Children's Nutrition Center in Little Rock, AR, have been studying the health effects of soyfoods, especially soy infant formula. Soy has been shown previously to reduce cholesterol. Since obesity and the adverse effects of cholesterol are being observed at younger ages each year, understanding how dietary factors work to control cholesterol could have significant implications for improved health management of the increasingly higher number of children in the United States population suffering from high cholesterol. Using the young rat as a model, we have identified the potential major mechanism by which soy reduces cholesterol. Soy reduces cholesterol by altering metabolism and transport of cholesterol. 4b.List other significant accomplishments, if any. MILK PROTEIN PRODUCT MAY PREVENT DIABETES Type 2 diabetes is a growing concern in the United States and is becoming more prevalent in children, thereby a mechanism to control this is needed. Scientists at the Arkansas Children's Nutrition Center in Little Rock, AR, have shown that a processed milk protein (partially hydrolyzed whey protein) reduces circulating insulin. This may be of great importance, since insulin resistance and type 2 diabetes is a growing health problem in the United States. The potential of having a dietary mechanism by which this could be managed would be very important for the general public. 4c.List any significant activities that support special target populations. none. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. Over the life of this research project, scientists of the Arkansas Children's Nutrition Center in Little Rock, AR, have: In animal studies, determined that it is possible to reduce the incidence of the two cancers responsible for most American cancer deaths (breast and colon) through diet alone. To accomplish this, we assembled a team of nutritionists, pharmacologists, toxicologists, cell biologists, molecular biologists, endocrinologists, immunologists, chemists, and animal and plant scientists to address the health effects of dietary factors, especially phytochemicals, during development. Established animal models to study dietary prevention of breast and colon cancer, and cardiovascular disease. Conducted studies in infants to confirm body composition, metabolism, and behavioral effects of infant feeding regimens. Developed new techniques to detect and quantitate food phytochemicals in human blood, tissues, urine, and foods. Defined the dynamics (pharmacokinetics, bioavailability, etc.) of soy isoflavones, the major phytochemicals found in infant formula. Determined that the major enzyme, CYP3A, that metabolizes 80% of all medications used in human medicine is significantly altered in rats fed diets made from the soy protein isolate used in soy infant formula. This could mean that the efficacy of important drugs (like antibiotics) used in children fed these diets would be lower, having profound implications for the health outcomes of illnesses that require antibiotic treatment. Used the Oxygen Radical Absorbance Capacity (ORAC) assay as a tool for (a) antioxidant capacity assessment of various foods and for standardization of nutritional supplements, and (b) evaluation of the effects of maturity at harvest and genetic background on antioxidant capacity in blueberries and strawberries. Evaluated variability in antioxidant capacity of commercially available antioxidant nutritional supplements and determined that quality control is needed to standardize commercial antioxidant supplements. Demonstrated increased antioxidant capacity in plasma following consumption of additional servings of fruits and vegetables or following a meal containing high antioxidant foods, allowing us to predict dietary intakes that may impact health outcomes. Determined the timing in the lifecycle in which soy protein isolate, rice protein isolate, and whey protein hydrolysate have significant health effects is of extreme nutritional importance. Determined that very early feeding of soy activates the genes for enzymes that are responsible for metabolism of medications and for activation and inactivation of toxic substances. These data have potentially important medical ramifications because they suggest that soy formula may alter the drug levels of children treated with medications. Taken the first steps in the development of two new animal models of human atherosclerosis [a disease that is initiated during pregnancy and early infancy] to better understand this disease. These models will allow researchers to identify which dietary factors can prevent cardiovascular disease, under what circumstances, and by what mechanisms. The questions being addressed in this project are very important and consistent with the National Program - 107 goals. This research directly supports ARS Strategic Plan Goal #4, Improve the Nation's Nutrition and Health. 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? The use of the ORAC procedure for the assessment of antioxidant capacity in fruits and vegetables and nutritional supplements is being implemented in industry. We have been working with the frozen food industry to develop more health-responsible foods. We have helped secure the future commercial success of the Arkansas mushroom industry. 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). Newspapers/Magazines: ACNC scientists: gave radio interviews (KARN); had stories published in newspapers (St. Louis Dispatch, Arkansas Democrat/Gazette, Daily Health News, Washington Post, LA Times, Washington Times, Chicago Sun Times, Dallas Morning News); and articles in magazines (Parent, Cooking Light, Health, Science World, Science News, ATT World Net). Local Presentations: ACNC investigators and staff have given numerous talks and presentations in the local community, including: Psychiatry Rounds, Pharmacology/Toxicology; to the ACH Clinical Staff; 5-A-Day Health Fair; Arkansas Cancer Research Center; Baptist Health Fair; Healthy Kids Summit; the ACH Health Fair; Life Quest; National Park Commission; National Grain Sorghum Producers; Arkansas Children's Hospital Research Institute; and the Annual Capital Research Expo. We have given more than 22 tours through the ACNC for various groups, including: dietetic interns; students; parents, organizations, candidates and perspective subjects. National and International Meetings and Presentations: The ACNC research team has presented more than 30 presentations at national and international meetings. Workshops and Special Meetings: ACNC Investigators gave several invited talks and participated in several invited workshops on such issues as health effects of soy, milk proteins, fruits and rice, as well as other nutritional issues to the North Little Rock School District, Arkansas Dietetic Association, American Council on Fitness and Nutrition, American Heart Association, and Small Fruits Research Group. ACNC was represented as a member of an industry sponsored board to help prepare an FDA food health claim for soy's role in prevention of cancer. ACNC presented talks on the effects of soy formula to European health professionals in the United Kingdom. Review Publications Luo, H., Fang, N., Li, Q., Yu, S., Badger, T.M. 2004. Hydroxylated triterpene alcohol ferulateds from rice bran. Journal of Natural Products. 68(1):94-97. Cho, M.J., Howard, L.R., Prior, R.L., Clark, J.R. 2004. Flavonoid glycosides and antioxidant capacity of various blackberry, blueberry, and red grape genotypes determined by high-performance liquid chromatography/mass spectrometry. Journal of the Science of Food and Agriculture. 84(13):1771-1782. Huang, D., Ou, B., Prior, R.L. 2005. The chemistry behind dietary antioxidant capacity assays. Journal of Agricultural and Food Chemistry. 3(6):1841-1856. Prior, R.L., Joseph, J.A. 2005. Berries and fruits in cancer chemoprevention. In: Bagchi, D, Pruess, H. editors. Phytopharmaceuticals in Cancer Chemoprevention. 1st Edition. New York, NY: CRC Press. p. 465-479. Ou, B., Hampsch-Woodill, M., Flanagan, J., Deemer, E.K., Prior, R.L., Huang, D. 2002. Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe. Journal of Agricultural and Food Chemistry. 50(10):2772-2777. Kopper, R.A., Odum, N.J., Sen, M., Helm, R.M., Stanley, S., Burks, W.A. 2004. Peanut protein allergens: II. gastric digestion is carried out exclusively by pepsin. Journal of Allergy Clinical Immunology. 114(3):614-618. Hidestrand, M., Shankar, K., Ronis, M.J., Badger, T.M. 2005. Effects of light and dark beers on hepatic cytochrome p450 expression in male rats receiving alcoholic beverages as part of total enteral nutrition. Alcoholism: Clinical and Experimental. 29(5):888-895. Prior, R.L. 2004. Biochemical measures of antioxidant status. Topics in Clinical Nutrition. 19(3):226-238. Fariba, R., Hunt, J., Badger, T.M. 2005. Controlled substitution of soy protein for meat protein: effects on calcium retention, bone, and cardiovascular health indices in postmenopausal women. Journal of Clinical Endocrinology and Metabolism. 90(1):181-189. Stewart, R.J., Askew, E.W., Mcdonald, C.M., Metos, J., Jackson, W.D., Balon, T.W., Prior, R.L. 2002. Antioxidant status of young children: response to an antioxidant supplement. Journal of The American Dietetic Association. 102(11):1652-1657. Sillman, K., Parry, J., Kirk, L.L., Prior, R.L. 2003. Pycnogenol does not impact the antioxidant or vitamin c status of healthy young adults. Journal of The American Dietetic Association. 103(1):67-72. Selvaraj, P., Fifadara, N., Nagarajan, S., Cimino, A., Wang, G. 2004. Functional regulation of human neutrophil FC gamma receptors. Immunological Research. 29(1-3):219-230. Velarde, M.C., Parisek, S.I., Eason, R.R., Simmen, F.A., Simmen, R.C. 2005. The secretory leukocyte protease inhibitor (SLPI) gene is a target of epidermal growth factor receptor action in endometrial epithelial cells. Journal of Endocrinology. 184(1):141-151. Prior, R.L., Wu, X., Gu, L., Mckay, S. 2004. Characterization of anthocyanins and proanthocyanidins in some varieties of ribes, aronia and sumbucus and their antioxidant capacity. Journal of Agricultural and Food Chemistry. 52(26):7846-56. Prior, R.L., Wu, X. 2004. Anthocyanins. In: Coates, P., Coates, P.M., editors. Encyclopedia of Dietary Supplements. New York, NY: Marcel Dekker. p 840-849. Wu, X., Pittman, H.E., Prior, R.L. 2004. Pelargonidin is absorbed and metabolize differently than cyanidin after marionberry consumption in pigs. Journal of Nutrition. 134(10):603-2610. Stewart, B., Ferguson, M., Badger, T.M., Nagarajan, S. 2005. Dietary soy protein isolate (SPI+) inhibits development of atherogenesis in familial hypercholesterolemic mouse model. Journal of Federation of American Societies for Experimental Biology. 19(5):A1460. Nagarajan, S., Fifadara, N.H., Selvaraj, P. 2005. Signal specific activation and regulation of human neutrophil FC gamma receptors. Journal of Immunology. 174(9):5423-5432. Badger, T.M., Ronis, M.J., Simmen, R.C., Simmen, F.A. 2005. Soy protein isolate and protection against cancer. Journal of American College of Nutrition. 24(2):146S-149S. Simmen, F.A., Badger, T.M., Xiao, R. 2005. Dietary exposure to soy or whey proteins alters colonic global gene expression profiles during vat colon tumorigenesis. Molecular Cancer. 4(1):1. Eason, R.R., Velarde, M.C., Chatman, L., Till, R.S., Geng, Y., Ferguson, M., Badger, T.M., Simmen, R.C. 2004. Dietary exposure to whey proteins alters rat mammary gland proliferation, apoptosis, and gene expression during postnatal development. Journal of Nutrition. 134(12): 3370-3377. Dave, B., Eason, R.R., Till, R.S., Geng, Y., Velarde, M.C., Badger, T.M., Simmen, R.C. 2005. The tumor suppressor PTEN mediates the pro-apoptotic activity of dietary genistein on mammary epithelial cells: implications for mammary cancer protection. Proceedings American Association of Cancer Research. 46:2663. Simmen, R.C., Eason, R.R., Till, R.S., Chatman, L., Velarde, M.C., Geng, Y., Korourian, S., Badger, T.M. 2004. Inhibition of nmu-induced mammary tumorigenesis by dietary soy. Cancer Letters. 224(1): 45-52. Xiao, R., Ferguson, M., Badger, T.M., Simmen, F.A. 2005. Dietary prevention of azoxymethane (AOM)-induced intestinal cancers by whey protein hydrolysate (WPH): are aberrant crypt foci (ACF) suitable intermediate endpoint biomarkers for tumor occurrence? Proceedings American Association of Cancer Research. 46:5180. Linz, A.L., Xiao, R., Parker, J.G., Simpson, P.M., Badger, T.M., Simmen, F.A. 2004. Feeding of soy protein isolate to rats during pregnancy and lactation suppresses formatoin of aberrant crypt foci in their progeny's colons: interaction of diet with fetal alcohol exposure. Journal of Carcinogenesis. 3:14. Carter, J.A., Xiao, R., Badger, T.M., Simmen, F.A. 2005. Dietary whey protein hydrolysate (wph) suppresses circulating insulin and c-peptide levels and inhibits colon tumorigenesis. The FASEB Journal. 19(4):A77. Haley, R., Hidestrand, M., Shankar, K., Lumpkin, C.K., Yarberry, B., Badger, T.M., Ronis, M.J. Estradiol protects against ethanol-induced bone loss in female rats by preventing osteoclast activation. The Toxicologist. 84(S-1):354. He, L., Simmen, F.A., Ronis, M.J., Badger, T.M. Ethanol has biphasic effects on expression of sterol regulatory element binding protein-1 (srebp-1) in rat fgc-4 hepatoma cells with inhibition at high doses resulting in induction of ADH class I. The Toxicologist. 84(S-1): 392. Ronis, M.J., Badger, T.M., Lumplin, C.K., Arnson, J., Hidestrand, M., Shankar, K., Haley, R. 2005. Mechanisms of ethanol-induced bone loss differ with physiological state. The Toxicologist. 84(S-1):211. Guacher, E.A., Graddy, L.G., Li, T., Simmen, R.C., Simmen, F.A., Schreiber, D.R., Liberles, D.A., Janis, C.M., Benner, S.A. 2004. The planetary biology of cytochrome p450 aromatases. BMC Biology. 2(1):19. Gu, L., Prior, R.L., Fang, N., Ronis, M.J., Clarkson, T.B., Badger, T.M. 2005. Interspecies differences of isoflavone metabolic phenotypes in female rats, pigs, monkeys and humans. The FASEB Journal. 19(4):A446. Gu, L., Prior, R.L., Fang, N., Ronis, M.J., Badger, T.M. 2005. Organ and subcellular distribution of genistein and diadzein in rats. The FASEB Journal. 19(4):A447. Wu, X., Pittman, H.E., Mckay, S., Prior, R.L. 2005. Anthocyanins (acns) were absorbed and metabolized differently in weanling pigs after feeding black currant (bc) or chokeberry (cb). The FASEB Journal. 19(4):A415. Gu, L., Wu, X., Ou, B., Harnly, J., Prior, R.L. 2005. Procyanidin (pc) content and total antioxidant capacity (tac) of chocolate and cocoa products. The FASEB Journal. 19(5):A1032. Ronis, M.J., Chen, Y., Badger, T.M. 2005. 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