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? Diabetes is one of the leading causes of lost productivity and death, but the real causes of diabetes are not fully understood. For the majority of people, the most important factors in controlling diabetes are the foods and nutrients consumed. Additional research is urgently needed to define foods and nutrients that play key roles in chronic diseases. One essential nutrient that has been shown to alter the risk factors associated with type 2 diabetes and cardiovascular diseases is chromium. Dietary intake of chromium is an important human health concern since total intake of chromium is suboptimal and diets with high amounts of simple sugars, which are common in most Westernized diets, facilitate chromium loss. There are also other insulin potentiating compounds found in plants that may improve insulin activity and prevent or alleviate glucose intolerance and diabetes. The objectives of our research are: 1. Elucidate the role of chromium in the onset of impaired glucose metabolism, using a stress induced rat model for chromium deficiency; 2. Elucidate methods to assess chromium status and to determine its roles in human nutrition; 3. Elucidate the role and mechanistic effects of insulin potentiating polyphenols from cinnamon on intracellular signals that regulate glucose uptake and oxidative stress; and 4. Define the safety and antioxidant effects of chromium, cinnamon and other naturally occurring insulin potentiating factors. These objectives address the National Program Action Plan to "adapt current methods or develop new methods to identify specific development enhancing and disease preventing bioactive factors and elucidate their mechanisms of action," and "develop an appropriate animal model that will be used to identify specific biomarkers that are associated with developmental effects or disease prevention." Our efforts to isolate and characterize natural products that improve glucose tolerance and reduce the risks of diabetes in the diverse population address the national priority to "characterize substances thought to benefit health and measure effects of these substances on biomarkers of developmental effects and illnesses of chronic diseases". 2.List the milestones (indicators of progress) from your Project Plan. Milestone 1 (12 - 48 months)- Determine an animal model to study the nutritional roles of chromium in nutrition. Ascertain conditions for chromium deficiency. Raise chromium deficient animals and treat with dexamethasone and optimize conditions. Analyze samples to verify chromium deficiency. Determine the role of chromium in human nutrition and methods to assess status. Verify collections and check for chromium contamination. Enroll subjects and conduct clinical measurements. Conduct measurements to ascertain chromium status. Milestone 2 (12 - 36 months)- Determine the roles of chromium and related variables in response to stress. Initiate collaborative arrangements and define conditions to determine the effects of stress on rats subjected to thermic stress. Subject animals to thermic stress and collect tissues and body fluids. Analyze samples to determine trace metal and other dietary changes that may be useful in combating stress. Milestone 3 (12 - 36 months)- Determine the role of cinnamon and other natural products in the control of glucose and insulin metabolism. Define conditions to monitor biological effects of insulin potentiating compounds in animals and in cell culture systems. Milestone 4 (24 - 48 months)- Conduct human studies to show that effects observed in animals and cell culture systems are relevant to humans. Measure effects on specific cell proteins and effects on nucleic acids. Milestone 5 (24 - 48 months)- Determine the antioxidant effects of cinnamon and other natural products. Measure effects in cell culture systems and protective effects on ultraviolet radiation. Milestone 6 (24 - 48 months)- Determine the safety of chromium, cinnamon and other natural products that improve insulin function. Measure effects on cell cycle, viability and deoxyribonucleic acid (DNA) degradation. Milestone 7 (48 - 60 months)- Complete technology transfer and recommendations for adequate doses of chromium and natural products of cinnamon that improve insulin function. 4a.What was the single most significant accomplishment this past year? Dietary intake of trace elements is often suboptimal and stress has negative effects on trace metal status. The trace elements, chromium, copper, selenium and zinc are often suboptimal in diets as normally consumed in the U.S. and most other countries. Suboptimal intakes of these nutrients are often associated with chronic diseases such as diabetes, cardiovascular diseases, premature aging and related diseases. Dietary status of these nutrients is often impacted negatively by stress. In conjunction with scientists from the French Department of the Military and the Josef Fourier University, Grenoble, France, we conducted a series of experiments to show that the response to stress may be negatively impacted by suboptimal intake of trace elements. In response to the stress of burn injury, copper and zinc were redistributed to the liver, selenium to the kidney and chromium to the muscle. These metals also function in the antioxidant protective mechanisms of the body and there were significant decreases in the activities of the associated protective antioxidant enzymes and metabolites. These studies demonstrate that improved trace metal status may be important in combating stress. 4b.List other significant accomplishments, if any. The antioxidant effects of vitamin C are well established, but vitamin C may also function as a pro-oxidant. In conjunction with scientists from Wisconsin and Josef Fourier University, Grenoble, France, we administered a standard ethylenediaminetetraacetic acid (EDTA) chelation cocktail solution with or without 5g of sodium ascorbate intravenously to five subjects. Oxidative stress markers including plasma malondialdehyde (MDA), protein thiol groups, total and oxidized glutathione, deoxyribonuclic acid (DNA)damage and antioxidant enzymes were measured for each subject before and after each therapy session. One hour following the intravenous intake of vitamin C, there were highly significant pro-oxidant effects on lipids, proteins and DNA associated with decreased activities of red blood cell (RBC) glutathione peroxidase and superoxide dismutase. For example, plasma MDA increased 29%, DNA damage, increased 38% and total thiol groups decreased 18%. Under the same conditions but without added vitamin C, there were no signs of oxidative damage. These data demonstrate that vitamin C administered intravenously may display strong pro-oxidant effects. The amount of vitamin C given intravenously should be closely monitored and the antioxidant/pro-oxidant effects should be established before inclusion of vitamin C in intravenous solutions. 4c.List any significant activities that support special target populations. People with glucose intolerance or diabetes can be a target of improved nutrition to help prevent and/or control diabetes; minority populations in the U.S. where economic and social factors limit appropriate food choices are especially vulnerable. There is also a need for preventative strategies in developing countries where people may not have access to (or cannot afford), therapy including drugs. Our studies demonstrate that chromium intake of people from Tunisia displayed large seasonal variations which may be related to specific ethnic foods. We have not observed this variation in subjects from the U.S. Our work with people with diabetes demonstrate that small dietary changes such as adding roughly one-half teaspoon of cinnamon per day to the diet can lead to significant improvements in risk factors associated with diabetes and cardiovascular diseases. 4d.Progress report. We have shown previously that supplemental chromium improves blood glucose, insulin and related variables of people with varying degrees of glucose intolerance. Recently, there have been some unsubstantiated reports that supplemental chromium may have toxic effects. We demonstrated that at intakes of chromium almost 30-fold above the adequate intake recently established by the Institute of Medicine there were no signs of toxicity and only benefits in human subjects. Studies just complete demonstrate that the toxicity of chromium is very low and that the chromium complexes used as nutrient supplements actually have protective effects on DNA in intact cells rather than increases DNA degradation. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. Type 2 diabetes and cardiovascular diseases are major contributors to poor health status of the U.S. population and a major drain on the health care system. Simple and inexpensive adjustments to diet that improve insulin function and lead to reductions in cardiac pathologies would have significant impact on the broad spectrum of the population where health care opportunities are limited. Our research has demonstrated that the risk factors associated with type-2 diabetes and cardiovascular diseases can be alleviated by compounds that improve insulin function, including chromium and other insulin potentiating factors found in foods, herbs and spices. Modern diets that are high in fats, sugars and other refined components are also low in chromium and other potentiators of insulin action. In addition, consumption of these foods increases chromium losses and this, coupled with a low intake of foods high in factors that improve insulin function, may lead to increased risk factors of diabetes and cardiovascular diseases, including elevated blood sugars and fats. These factors have significant negative effects on health and are likely related to the large recent increases in the incidence of type 2 diabetes mellitus and obesity. We have expanded our collaborative studies that resulted in a joint Israel-U.S. Binational Agricultural Research and Development (BARD) proposal entitled, "Spices, Medicinal Plants and Yeasts as Dietary Sources for Antioxidant, Lipid Lowering and Hypoglycemic Materials," to address the need to find biologically active compounds in foods that can improve efficient glucose utilization. The early work in the project has lead to a joint human studies project entitled "Chromium and Insulin Action" that is funded by the National Institutes of Health (NIH); these studies are designed to determine the mode of action of chromium and methods to determine chromium status in humans. Earlier studies in animal models to examine chromium deficiency and to elucidate its mode of action will be expanded during the life of the current project. This year we expanded or work with an additional Cooperative Research and Development Agreement (CRADA) partner to improve the polyphenols found in cinnamon and monitor their activity and safety in cell culture, animal and human studies. The impact of the work will provide important information on the nutritional needs of the diverse U.S. population, and on trace element nutrition to determine requirements, bioavailability, and health consequences of suboptimal nutrient intakes. 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 benefits of our work have already been transferred to the general population in that millions of people are improving their chromium nutrition, due in part to our work. Our patent entitled, "Chromium Histidine Complexes as Nutrient Supplements" was issued February 10, 2004, and several companies have expressed interest in licensing. Our patent entitled "Insulin Potentiating Compounds from Cinnamon," S.N.10/046,088, D.N. 0078.02, is close to being issued, and has already led to a cooperative research agreement and has also led to funding of a human study to substantiate our previous study and two additional cooperative human studies. We are currently negotiating an additional multi-year cooperative research agreement to extend this research to use naturally occurring compounds from cinnamon as food additives. Since these compounds improve the action of insulin and, therefore, control blood sugar, it is postulated that adding these compounds to foods high in sugar will lead to decreased negative effects associated with eating high sugar foods. 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). A summary of the cinnamon research has been added to the Beltsville Area Web site, http://www.ars.usda.gov/is/pr/2004/040419.htm. Cinnamon articles have been the subject of numerous newspaper and magazine articles and an Internet search of our work leads to hundreds of articles in the lay press. Review Publications Hininger, I., Waters, R., Osman, M., Garrel, C., Fernholz, K., Roussel, A.M., Anderson, R.A. 2005. Acute prooxidant effects of vitamin c in EDTA chelation therapy and long term antioxidant benefits of therapy. Free Radicals in Biology and Medicine. 38:1565-1570. Anderson, R.A., Polansky, M.M. and Bryden, N.A. 2004. Stability and absorption of chromium and absorption of chromium histidinate complexes by humans. Biological Trace Elements Research. 101:211-218. Schoene, N.W., Kelly, M.A., Polansky, M.M., Anderson, R.A. 2005. Water soluble polymeric polyphenols from cinnamon inhibit proliferation and alter cell cycle distribution patterns of hematologic tumor cell lines. Cancer Letters. 230:134-140.