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? This CRIS consists of the Bone Metabolism Laboratory and the Vitamin K Laboratory where scientists are working to improve the understanding of how diet impacts bone health in older men and women. Serum 25-hydroxy vitamin D [25(OH)D] is the best clinical measure of vitamin D status. The Bone Metabolism Laboratory is working to define factors that influence the 25(OH)D concentrations achieved at a given vitamin D intake level in the wintertime. Scientists are conducting a study of the effect of body mass index on the utilization of dietary vitamin D in 389 men and women age 65 and older who participated in a placebo-controlled calcium and vitamin D intervention study. They will also test the hypothesis that the increase in serum 25(OH)D in response to oral dosing with vitamin D is blunted at low calcium intakes. This will be studied in a new randomized controlled trial in which 800 IU/day of vitamin D will be given to older subjects with high and low calcium intakes. Scientists will also examine the impact of endogenous gonadal-hormone levels on 25(OH)D responses to supplemental vitamin D. Falls are a common source of serious injury, including fractures, in older subjects. Recent evidence suggests that low vitamin D levels may contribute to muscle weakness and increased risk of falling in this population. Scientists will examine this possibility in the 389 older men and women who participated in a 3-year calcium and vitamin D intervention study. The impact of lean body mass, gonadal hormone levels, and usual activity levels on risk of falling will also be examined. These vitamin D studies will improve the scientific basis for determining the vitamin D intake requirements for older men and women. A second research initiative of the Bone Metabolism Laboratory is to define the impact of dietary protein and the acid-base balance of the diet on bone health. Several small protein intervention studies will be conducted to define the relative roles of protein, added aromatic or branched-chain L-amino acids, calcium, vitamin D, and alkaline foods (fruits and vegetables) or supplements, such as potassium bicarbonate, on short-term indices of bone metabolism including biochemical markers of bone turnover, calcium absorption, urinary calcium excretion, and insulin-like growth factor-1. A supplement (expected to contain protein, calcium, vitamin D, alkali, and perhaps added branched-chain amino acids) will be tested in a large longitudinal intervention trial in older men and women to determine its impact on rates of change in bone mineral density (BMD). This research falls under National Program 107 - Human Nutrition, and is related to component 4: Nutrient Requirements. The ability to establish dietary recommendations for vitamin K is limited by an inadequate understanding of the dietary sources and metabolic fate of phylloquinone from the diet. Studies are being conducted to identify the forms, amounts and biological activity of vitamin K in the food supply. Vitamin K food composition data generated from food analysis will be incorporated into the national nutrient databases for the purpose of menu design, dietary assessment, and national intake surveys. Stable isotopes will be used to elucidate the absorption and transport of vitamin K from vegetables, the primary dietary source of vitamin K in the United States (US). Information is also being collected on the physiological significance of measures used to assess vitamin K nutritional status, with a focus on the impact of vitamin K supplementation on bone and vascular health. The largest of these studies is a 3-year randomized, placebo-controlled vitamin K intervention trial in 452 men and women, age 60-80, who are calcium and vitamin D replete. This study is currently underway, with all subjects enrolled. The main analysis will determine if there are bone density gains among those receiving vitamin K supplementation. Secondary analysis will determine if there is a reduction in the progression of age-related vascular calcification among those receiving vitamin K supplementation. This research falls under National Program 107 - Human Nutrition, program components 1: Composition of Foods, 2: Bioavailability of Nutrients and Food Components, and 4: Nutrient Requirements. 2.List the milestones (indicators of progress) from your Project Plan. Bone Metabolism Laboratory Objective 1: Refine the approach to determining the vitamin D requirement for optimal musculoskeletal and dental health in men and women Objective 2: Define the impact of dietary protein and the acid/base balance of the diet on musculoskeletal health in men and women Objective 3: Define the impact of calcium and vitamin D intake on insulin resistance and risk of developing Type 2 diabetes in men and women Milestones: 2005 1. To determine the impact of body mass index on the serum 25(OH)D response to supplemental vitamin D. Objective 1 2. To determine the impact of calcium intake on the 25(OH)D response to supplemental vitamin D. Objective 1 3. To assess the long-term effect of vitamin D plus calcium on physical function and on the risk of falling in older men and women. Objective 1 2006 1. To determine the association between serum vitamin D levels and risk of gingivitis. Objective 1 2. To define and compare patterns of calcium and vitamin D supplement use in a random sample of women with and without osteoporosis. Objective 1 3. To determine whether the level of calcium intake influences any association between the ratio of nitrogen to potassium in the urine and rates of bone loss in older men and women. Objective 2 4. To determine the association between the ratio of nitrogen to potassium in the urine, an index of the acidogenicity of the diet, and 3-year rates of bone loss in healthy older men and women. Objective 1 5. To determine the impact of stimulating the calcium sensor receptor on gastric acid secretion and calcium excretion. Objective 2 6. To determine the association between intakes of calcium and vitamin D and incidence of developing Type 2 diabetes. Objective 3 7. To determine and compare the effects of selected aromatic and branched-chain amino acids on urinary calcium excretion in healthy older men and women. Objective 2 2007 1. To study the association between endogenous gonadal hormone levels and physical function and risk of falling in elderly men and women. Objective 1 2. Determine mechanisms by which vitamin D influences muscle strength and risk of falling. Objective 1 3. To determine whether alkalinization of the diet will attenuate the urine calcium losses induced by high protein diets. Objective 2 4. To determine the effect of supplementation with calcium and vitamin D on fasting insulin and glucose levels in healthy older men and women. Objective 3 2008 1. Define mechanisms by which specific amino acids affect intestinal calcium absorption. Objective 2 2. To begin a randomized, controlled longitudinal trial to determine whether alkalinization of the diet will lower the bone turnover rate, and thus lower risk of fracture, in healthy men and women who are being supplemented with calcium and vitamin D. Objective 2 2009 1. Continue the randomized, controlled longitudinal trial to determine whether alkalinization of the diet will lower the bone turnover rate, and thus lower risk of fracture, in healthy men and women who are being supplemented with calcium and vitamin D. Objective 2 Vitamin K Laboratory Objective 1: Identify and measure the amounts of individual dietary forms of vitamin K in key foods representative of the national food supply, and estimate their relative contribution to total dietary vitamin K intake. Objective 2: Define endogenous and exogenous factors affecting bioavailability of phylloquinone, the major form of vitamin K in circulation. Stable isotope techniques will be developed to measure phylloquinone in individual lipoprotein fractions in a fasting and fed state. These stable isotope techniques will be used to determine if age, lipid profile and current vitamin K nutritional status influence the absorption and transport of phylloquinone in younger and older men and women. Apolipoprotein E genotype will be examined as a potential influence on vitamin K nutritional status in men and women. Objective 3: Determine the impact of vitamin K supplementation, in an amount that is expected to be nutritionally optimal and safe, on bone, joint and vascular health in elderly men and women. Milestones: 2005 1. Characterize and quantify multiple forms of vitamin K in food samples representative of the US food supply. Objective 1 2. Create an expanded, yet targeted vitamin K database using the aforementioned food analysis data, to be used for a food frequency questionnaire. Objective 1 3. To assess the dietary and non-dietary determinants of circulating levels of dihydrophylloquinone, a form of dietary vitamin K, in the Framingham Heart Study. Objective 1 4. To develop stable isotope techniques using liquid chromatography mass spectrometry (LC/MS) to measure vitamin K absorption and transport. Objective 2 2006 1. Quantify multiple forms of vitamin K in dietary supplements representative of the US food supply. Objective 1 2. To relate the dietary intakes of dihydrophylloquinone, a form of dietary vitamin K, to physiological outcomes, such as bone health, in the Framingham Heart Study. Objective 2 3. To assess the role of apolipoprotein E genotype in modulating vitamin K status in the Framingham Heart Study. Objective 2 4. To develop a rat model to examine the effects of vitamin K supplementation on the transport of vitamin K to various tissues. Objective 2 2007 1. To determine the impact of vitamin K supplementation, in an amount that is expected to be nutritionally optimal and safe, on bone, joint and vascular health in 452 elderly men and women. Objective 3 2. To estimate the usual dietary intakes of menaquinone-4, a form of vitamin K present in animal products, among men and women participating in the Framingham Heart Study. Objective 1 2008 1. To complete enrollment of a bioavailability study in younger and older men and women to examine the role of age, lipid profile and current vitamin K status on absorption and relative bioavailability of vitamin K from a vegetable. Objective 2 2. To conduct a cross-sectional analysis of phylloquinone, dihydrophylloquinone and menaquinone-4 intakes and plasma levels in the baseline data from 452 elderly men and women participating in a vitamin K supplementation trial to provide information on the dietary and non-dietary determinants of vitamin K that are specific to the elderly. Objective 2 2009 1. To examine the roles of apolipoprotein E, vitamin K epoxide reductase, and vitamin K-dependent gamma carboxylase genotypes in modulating vitamin K status among participants in a vitamin K supplementation trial. Objective 2 2. To determine the impact of vitamin K supplementation on measures of inflammation, as they relate to bone and vascular health. Objective 3 4a.What was the single most significant accomplishment this past year? Vitamin K and Osteoarthritis - Osteoarthritis is a common degenerative disease that increases in prevalence with age. In animal models, an inadequate amount of vitamin K has been associated with abnormal development of cartilage and excessive calcification in joints. Scientists in the Vitamin K Laboratory tested the hypothesis that adequate vitamin K status, as measured by intake and plasma levels, is associated with less hand and knee osteoarthritis in older men and women participating in the Framingham Heart Study compared to those with poor vitamin K status. Among the 1276 men and women who completed dietary assessment and had hand and knee radiographs, those with higher dietary intakes of vitamin K had less abnormal calcification in their hand joints compared to those with low dietary intakes of vitamin K. Among the 672 men and women who had vitamin K biochemical measures and hand and knee radiographs, individuals with higher plasma vitamin K levels had fewer joints with osteoarthritis and abnormal calcification compared to those with low plasma vitamin K levels. These data suggest that vitamin K may be a potentially modifiable factor in the prevention of this age-related form of arthritis. 4b.List other significant accomplishments, if any. Vitamin D, Calcium and Falls – Supplementation with vitamin D and calcium in frequently recommended amounts significantly lowered risk of falling in our cohort of 389 healthy older men and women. Relatively sedentary women appeared to have the greatest risk reduction with supplementation. Falls are the largest single cause of injury in the elderly and 90% of fractures result from falls. In a placebo-controlled calcium and vitamin D intervention trial, scientists in the Bone Metabolism Laboratory acquired information on fall rates every 6 months for 3 years and compared the fall rates among the supplemented and placebo groups. Broad-based increases in vitamin D and calcium intake were shown to have the potential to substantially reduce fractures and other injuries in the elderly by lowering the risk for falling. Vitamin K in Meats and Dairy – Menaquinone-4 is a form of vitamin K that is formed by conversion of phylloquinone, the primary form in the diet, in certain tissues. Menaquinone-4 may have a physiological role that is independent of the universal role of a cofactor that is attributed to all forms of vitamin K. Until now, there has been no systematic analysis of menaquinones in the US food supply. Scientists in the Viitamin K Laboratory analyzed the menaquinone-4 content of over 2,000 key foods collected as part of the National Food and Nutrient Analysis Program. This form of vitamin K was present in a variety of meats and dairy foods, although no single food item was identified as a rich dietary source of vitamin K. The presence of menaquinone-4 in meat and dairy foods could be important as physiologic functions unique to menaquinone-4 are identified. 4c.List any significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. As we complete the first year of this 5-year program cycle, the main accomplishment of the Bone Metabolism Laboratory is finding that supplementation with calcium and vitamin D lowers risk of falling in older subjects. Falls are common occurrences in the elderly – 30% of subjects age 65 and 40 to 50% of those age 80 will fall in one year and falls are a major precipitant of fractures. Dissemination of this finding in the scientific and medical community should lead to increases in vitamin D and calcium intakes and a reduction in falls and fractures in older men and women. This finding is related to Milestone #3: To assess the long-term effect of vitamin D plus calcium on physical function and on the risk of falling in older men and women, and to National Program 107 - Human Nutrition program component 4: Nutrient Requirements, and ARS Strategic Plan Performance Measure 4.1.2.: Improve Human Health by Better Understanding the Nutrient Requirements of Individuals and the Nutritional Value of Foods. The major accomplishment of the Vitamin K Laboratory has been to demonstrate that high dietary intakes and biochemical measures of vitamin K are associated with a reduced risk for certain chronic diseases associated with abnormal calcification in older men and women. Of concern is the consistent observation that dietary intakes of vitamin K are lower among U.S. adults than previously assumed. This finding is related to our FY 2007 milestone 1, which is to determine the impact of vitamin K supplementation in an amount that is expected to be nutritionally optimal and safe, on bone, joint and vascular health in 452 elderly men and women, and to National Program 107 - Human Nutrition, program component 4: Nutrient Requirements, and ARS Strategic Plan Performance Measure 4.1.2.: Improve Human Health by Better Understanding the Nutrient Requirements of Individuals and the Nutritional Value of Foods. 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 Bone Metabolism Laboratory’s finding that the calcium intake does not influence the increase in serum 25(OH)D that occurs with vitamin D supplementation (Milestone. 2)has been presented at a scientific meeting and published in the peer-reviewed literature. This information will be useful in determining the calcium and vitamin D requirements. No intellectual property rights issues apply. The research findings of the Vitamin K Laboratory have been presented to the scientific community at a number of meetings and have also been published in prestigious peer-reviewed scientific journals. Results of this research reach the public directly through the NIH National Resource Center on Osteoporosis and Related Bone Diseases at the National Osteoporosis Foundation, through press releases, print and electronic media, and the web. Vitamin K food composition data were electronically transferred to ARS scientists at Beltsville USDA Nutrient Data Laboratory for input into the USDA Nutrient Databank System. These data will be used by scientists doing nutrition-related research, by the food industry, and by consumers. 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). Journal of American Medical Association – Better osteoporosis management a priority (describing Dr. Dawson-Hughes’ views on vitamin D and Bone health) - May 25, 2005. Nutrition Action magazine – Bonnie Liebman – Breaking up: strong bones need more than calcium - April 2005. Balance Tufts Daily Newspaper – Erica Nakamota – Nutrigenomics studies how DNA individualizes our diet - April 27, 2005. US News World Report – National Issues Briefing, Bone Health and Osteoporosis (Calcium and vitamin D in the diet) - October 20, 2004. Review Publications Pro-Risquez, A., Harris, S.S., Song, L., Rudicel, S., Barnewolt, B., Dawson-Hughes, B. 2004. Calcium supplement and osteoporosis medication use in women and men with recent fractures. Osteoporosis International. 15:689-694. Bischoff-Ferrari, H.A., Dietrich, T., Orav, J.E., Hu, F.B., Zhang, Y., Karlson, E., Dawson-Hughes, B. 2004. Higher 25-hydroxyvitamin D levels are associated with better lower extremity function in both active and inactive adults 60+ years of age. American Journal of Clinical Nutrition. 80:752-758. Booth, S.L., Broe, K.L., Peterson, J.W., Cheng, D.M., Dawson-Hughes, B., Gundberg, C.M., Cupples, L.A., Wilson, P.W., Kiel, D.P. 2004. Associations between vitamin k biochemical measures and bone mineral density in men and women. Journal of Clinical Endocrinology and Metabolism. 89(10):4904-4909. Hyten, D.L., Song, Q., Costa, J., Cregan, P.B. 2003. Linkage disequilibrium in four soybean populations [abstract]. BARC Poster Day. Abstract 14. Vazquez, E., Song, L.Y., Dawson-Hughes, B. 2004. Precision of bone mineral density scans at the proximal tibia in osteoarthritic subjects. Journal of Clinical Densitometry. 7(4):364-367. Bischoff-Ferrari, H.A., Willett, W.C., Wong, J.B., Giovannucci, E., Dietrich, T., Dawson-Hughes, B. 2005. Fracture prevention by Vitamin D supplementation: A meta-analysis of randomized controlled trials. Journal of the American Medical Association. 293:2257-2264. Dawson-Hughes, B. 2004. Dietary protein and the skeleton. In: Burckhardt, P., Dawson-Hughes, B., Heaney, R.P. editors. Nutritional Aspects of Osteoporosis. 2nd edition. San Diego, CA.: Elsevier, Inc. p. 399-409. Bischoff, H.A., Dawson-Hughes, B. 2004. Defining optimal 25-hydroxyvitamin D levels in younger and older adults based on hip bone mineral density. In: Burckhardt, P., Dawson-Hughes, B., Heaney, R.P. editors. Nutritional Aspects of Osteoporosis. 2nd edition. San Diego, CA: Elsevier, Inc., p. 235-244. Dawson-Hughes, B., Heaney, R.P., Holick, M., Lips, P., Meunier, P.J., Vieth, R. 2004. Vitamin D round table. In: Burckhardt, P., Dawson-Hughes, B., Heaney, R., editors. Nutritional Aspects of Osteoporosis. 2nd edition. San Diego, CA.: Elsiever, Inc. p. 263-270. Dawson-Hughes, B. 2004. Racial/ethnic considerations in making recommendations for vitamin D for adult and elderly men and women. In: Proceedings of the NIH Vitamin D Conference, October 10, 2003. American Journal of Clinical Nutrition. 80(Suppl):1763S-1766S. Dawson-Hughes, B., Heaney, R.P., Holick, M.F., Lips, P., Meunier, P.J., Vieth, R. 2005. Estimates of optimal vitamin D status. Osteoporosis International [on-line]. Available at http://www.springerlink.com Dawson-Hughes, B. 2005. The role of vitamin d in fracture prevention. BoneKEy-Osteovision. 2(4):6-10. (Epub) Available at http://www.bonekey-ibms.org/cgi/content/full/ibmske;2/4/6 DOI: 10.1138/20050156 Wilsey, B.J., Polley, H.W. 2004. Realistically low species evenness does not alter grassland species-richness-productivity relationships. Ecology. 85(10):2693-2700. Jajoo, R., Song, L., Ramussen, H., Dawson-Hughes, B. 2004. Effect of change in dietary acid-base balance on bone turnover and urine calcium excretion. [abstract] Journal of Bone and Mineral Research. 19(Suppl 1):S448-449. Goussous, R.Y., Song, L., Dallal, G., Dawson-Hughes, B. 2004. The effect of calcium intake on the vitamin D requirement. [abstract] Journal Bone and Mineral Research. 19(Suppl 1):S202. Bischoff-Ferrari, H.A., Orav, J.E., Dawson-Hughes, B. 2004. Effect of vitamin D3 plus calcium on fall risk in older men and women: a 3-year randomized controlled trial. [abstract] Journal of Bone and Mineral Research. 19(Suppl 1):S57. Booth, S.L., Golly, I., Sacheck, J.M., Roubenoff, R., Dallal, G.E., Hamada, K., Blumberg, J.B. 2004. Effect of vitamin E supplementation on vitamin K status in adults with normal coagulation status. American Journal of Clinical Nutrition. 80:143-148. Schulze, K.J., O'Brien, K.O., Germain-Lee, E.L., Booth, S.L., Leonard, A., Rosenstein, B.J. 2004. Bone calcium deposition is associated with availability of dietary calcium and cirulating leptin and osteocalcin concentrations in girls with cystic fibrosis. Journal of Clinical Endocrinology and Metabolism. 89(7)3385-3391. Cury, R.C., Ferencik, M., Hoffmann, U., Ferullo, A., Moselewski, F., Abbara, S., O'Donnell, C.J., Booth, S.L., Brady, T.J., Achenbach, S. 2004. Epidemiology and association of vascular and valvular calcium quantified by multi-detector computed tomography in elderly asymptomatic subjects. American Journal of Cardiology. 94:348-351. Braam, L.A., Mckeown, N.M., Jacques, P.F., Lichtenstein, A.H., Vermeer, C., Wilson, P.W., Booth, S.L. 2004. Dietary phylloquinone intake as a potential marker for a heart-healthy dietary pattern in the Framingham Offspring Cohort. Journal Of The American Dietetic Association. 104(9):1410-1414. Damon, M., Zhang, N., Haytowitz, D.B., Booth, S. 2005. Phylloquinone (Vitamin K1) content of vegetables. Journal of Food Composition and Analysis. 18:751-758. Available online at http://www.sciencedirect.com doi:10.1016/j.jfca.2004.07.004 Erkkila, A.T., Lichtenstein, A.H., Jacques, P.F., Hu, F.B., Wilson, P.W., Booth, S.L. 2005. Determinants of plasma dihydrophylloquinone in men and women. British Journal of Nutrition. 93:701-708. Goussous, R., Song, L., Dallal, G.E., Dawson-Hughes, B. 2005. Lack of effect of calcium intake on the 25-hydroxyvitamin D response to oral vitamin D3. Journal of Clinical Endocrinology and Metabolism. 90(2): 707-711.