Progress 01/18/09 to 01/17/14
Outputs
Progress Report Objectives (from AD-416): Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti- inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. Objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. Objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/ obese individuals. Objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. Ojective 2B. Examine anti-inflammatory activities of polyphenol-rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. Objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of B-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. Objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. Objective 3B. Estimate the impact of daily consumption of food sources of CX or B-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. Objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. Objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. Objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Objective 5: Identify the role of dietary selenium and selenoproteins in regulating cellular responses to oxidative stress. Objective 5A. Identify the pro-inflammatory and anti-inflammatory proteins S-glutathionylated by selenoprotein W. Objective 5B. Determine the role of selenoprotein W in key inflammatory pathways. Approach (from AD-416): The impact of selenium, vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. The effect of selenium on cell division and cell signaling will be examined in cell culture. This is the final report for this project which expired in January of 2014 and has been replaced by the new project, 5306-53000-001-00D, "Assessing the Impact of Diet on Inflammation in Healthy and Obese Adults in a Cross-Sectional Phenotyping Study and a Longitudianal Intervention Trial". Please see the report for the new project for additional information. Objective 1: During the current year laboratory analysis was substantially completed on this project and data analysis began. Over the entire project period we successfully validated a method for use in human volunteers to characterize the pattern of vaccine-specific cytokine production in response to tetanus toxoid vaccination. That study resulted in one publication. Results of the vitamin D intervention trial, using these methods, will result in a second publication during the coming year. Results will address the question of whether daily doses of vitamin D in excess of that needed to maintain bone health and calcium metabolism are needed to maintain immune function, as assessed by a T-cell mediated immune response to tetanus toxoid vaccination (among other endpoints). In addition, work with collaborating scientists on sibling projects continues. One study is examining the effects of vitamin D supplementation on inflammation during pregnancy. This randomized intervention trial has been completed and a manuscript is in preparation. Work continues on additional human studies with the NIH-funded Adolescent Trials Network examining the effect of vitamin D supplementation on vitamin D status, markers of systemic inflammation, bone density and kidney function in adolescents and young adults with HIV infection. Objective 2: (2A) During the current year isolated lymphocytes were treated with physiological levels of resveratol glucuronide, the major metabolite of the plant antioxidant resveratrol, to define how this metabolite alters gene expression and function of these immune cells. Next generation sequencing revealed multiple genes that were differentially expressed in the lymphocytes treated with the glucuronide metabolite compared to the untreated cells. The confirmation analysis of differentially expressed genes is in progress. (2B) The U.S. High Bush Blueberry Council awarded a collaborative grant to study whether feeding humans blueberries will decrease the inflammatory response to a fatty meal. This study is complete, data analysis has been completed and a manuscript describing results is in preparation. (2C) Mouse studies completed in 2010 showed that feeding purified polyphenols at levels higher than typically found in foods did not kill leukemia cells. The diet studies planned for this year with polyphenol-rich foods were not pursued as this approach is likely to be ineffective. With regard to overall progress during the funding period, two human trials were completed to determine the anti-inflammatory and lipid lowering effects of dietary strawberries and grapes in obese individuals. Both fruits reduced lipid particles associated with cardiovascular disease. Furthermore, it was shown that dietary intake of these two fruits was able to sensitize monocytes to a bacterial trigger, suggesting that fruit intake can increase sensitivity of this immune cell population to infectious agents in the obese who are at a higher risk for developing infections compared to normal weight individuals. Three manuscripts were prepared and published with these data. Mouse studies were conducted to explore the anti-inflammatory effects of strawberries and one manuscript was published with these data. Multiple mouse studies were conducted to determine if the plant polyphenols resveratrol (found in grapes, blueberries, and peanuts) and curcumin (found in the spice turmeric) could prevent a type of infant leukemia that has a poor prognosis in human infants and children. Four manuscripts were published with these data. Other studies were performed in cell culture systems to evaluate the anti-leukemia effects of several plant-derived chemicals, including quercetin and kaempferol found in many fruits and vegetables, and parthenolide found in the herb feverfew. Three manuscripts were published with these data. Metabolites of resveratrol were evaluated for their effects on gene expression using isolated T lymphocytes and transcriptome sequencing technology in order to discover mechanisms of action of the polyphenol metabolites that are present in vivo. These experiments are in progress. Objective 3: During the present year, whole body metabolism of beta- cryptoxanthin was measured in the Mongolian gerbil, the best small animal model of carotenoid metabolism. Beta-cryptoxanthin accumulated in most tissues, especially the liver, kidney, and adipose, which is similar to what occurs with another vitamin A-forming carotenoid, beta-carotene. Vitamin A (retinol and retinyl esters) was the most abundant product formed by beta-cryptoxanthin. There was no evidence of other metabolites (such as 3-hydroxy-vitamin A or vitamin A2), which might interfere with vitamin A metabolism. These results show that beta-cryptoxanthin-rich foods can be good sources of vitamin A, without accumulating similar products that might interfere with vitamin A metabolism. With regard to progress during the full funding period, work from this project demonstrated that beta-cryptoxanthin appears to be more bioavailable from its primary food sources than other vitamin A-forming carotenoids, such as beta-carotene and alpha-carotene. Once absorbed, beta-cryptoxanthin accumulated in most tissues, especially the liver, which is similar to other carotenoids. Vitamin A (retinol and retinyl esters) was the most common product formed from beta-cryptoxanthin, with no evidence that other metabolites which could interfere with vitamin A metabolism. Results from a human study showed that beta-cryptoxanthin was 4-fold more bioavailable than the beta-carotene and appeared in breast milk as well as serum. These results show that beta-cryptoxanthin-rich foods are promising sources of vitamin A. The effectiveness of biofortified cassava for increasing beta-carotene and vitamin A concentrations in ten healthy adult women was estimated during this project. Data from this human study were combined with cassava consumption data from countries in Africa where cassava is a staple food, to model the daily vitamin A intake of women in Africa. This work estimated that if biofortified cassava completely replaced traditional low-carotenoid, white cassava in the diet, it could meet recommended vitamin A intakes for most women (92-100%) in six African countries where it is a staple crop: Angola, Central African Republic, Congo, Ghana, Mozambique, and Nigeria. While this estimate is preliminary and requires validation, the work demonstrates that consumption of biofortified cassava, processed to maintain beta-carotene, may be able to increase vitamin A status in African populations and other areas of the world where cassava is a staple crop. Objective 4: While studies of vitamin A and immunity were not conducted in the mouse model as planned, work on an extramurally funded project (5306-51530-018-24T) with collaborating scientists evaluated the impact of high-dose vitamin A supplementation (or placebo) at birth on vaccine responses and other measures of T-cell function in 306 Bangladeshi infants. The trial has been completed and data analysis and manuscript preparation are in progress. In addition, work on a second extramurally funded project (5306-51530-018-63T) has begun in the same cohort of infants to determine if vitamin A supplementation affects establishment of intestinal microbiota through the first two years of life, and whether the microbial community is associated with vaccine responses. This work will continue for two more years. Objective 5: Work this year characterized the effects of selenoprotein W1 (SEPW1) depletion on the intracellular localization of the epidermal growth factor receptor, using markers for specific endosomal compartments. We characterized the effect of SEPW1 on endocytosis and intracellular trafficking of the epidermal growth factor and transferrin receptors. In- house produced and commercial antibodies were screened for their usefulness in detecting SEPW1 by indirect immunofluorescence microscopy in order to characterize the cellular localization of this selenoprotein. Unfortunately, none of the antibodies were suitable, perhaps due to the apparently low abundance of this protein. Over the entire project period substantial progress was made in characterizing the cellular activities of SEPW1. In brief, SEPW1 knockdown diminished SEPW1 protein levels and induced a G1 arrest in the cell cycle and this effect appears to be mediated by disruption of cell-surface receptor signaling (e.g., by the epidermal growth factor receptor and other related receptors) and is also dependent on expression of the protein p53, which plays a role in receptor-mediated signal transduction to the cellular nucleus. This work strongly suggests that SEPW1 is a required component of cellular responses triggered by extracellular receptors responding to growth factors, cytokines, hormones and related proteins. Significant Activities that Support Special Target Populations: The work of this project on vitamin D supplementation directly benefits historically underserved segments of the population in that national survey data from the USDA shows that vitamin D deficiency is more common in African American and Mexican American segments of the U.S. population. This work will provide a stronger scientific basis for dietary recommendations to these segments of the U.S. population. Accomplishments 01 Human lymphocyte functions are affected by resveratrol metabolites. ARS scientists working at the Davis, California location found that the major metabolites of resveratrol found in humans after ingestion, 2 resveratrol glucuronides and resveratrol sulfate, were not effective in killing human T cell leukemia cells in a cell culture system. However, resveratrol sulfate was able to disrupt mitochondrial membrane potential without killing the cells. These results suggest that resveratrol sulfate may be the more potent metabolite of resveratrol and may induce functional changes in human T lymphocytes. Further studies are in progress to define the mechanisms of action of this and other phytochemical metabolites generated in the body after consumption. These results are novel and will help determine how plant antioxidants affect human health by modulating the activity of immune cells. 02 Metabolism of beta-cryptoxanthin, a vitamin A-forming carotenoid. An ARS scientist working in Davis, California measured whole-body metabolism of beta-cryptoxanthin in the Mongolian gerbil, the best small animal model of carotenoid metabolism. Results showed that beta- cryptoxanthin accumulated in most tissues, especially the liver, kidney, and adipose. This is similar to another vitamin A-forming carotenoid, beta-carotene. Vitamin A (retinol and retinyl esters) was the most abundant product formed by beta-cryptoxanthin. There was no evidence of other metabolites (such as 3-hydroxy-vitamin A or vitamin A2), which might interfere with vitamin A metabolism. These results show that beta-cryptoxanthin-rich foods can be good sources of vitamin A, without accumulating similar products that might interfere with vitamin A metabolism.
Impacts
(N/A)
Publications
- Burri, B.J. 2014. The current impact and potential of biotechnology to improve the capacity of orange-fleshed sweet potato to prevent vitamin A deficiency. IN: Ramawat, K., Merriloon, J.M., editors. Bulbous Plants: Biotechnology. Boca Raton,FL: CRC Press. 287-310.
- Havens, P.L., Kiser, J.J., Stephensen, C.B., Hazra, R., Flynn, P.M., Wilson, C.M., Rutledge, B., Bethel, J., Pan, C.G., Woodhouse, L.R., Van Loan, M.D., Liu, N., Lujan-Zilbermann, J., Baker, A., Kapogiannis, B.G., Gordon, C., Mulligan, K. 2013. Higher plasma vitamin D binding protein and lower free calcitriol with tenofovir treatment: Cause of a functional vitamin D deficiency. Antimicrobial Agents and Chemotherapy. 57:619-28.
- Bornhorst, G., Roman, M., Rutherfurd, S.M., Burri, B.J., Singh, R., Moughan, P.J. 2013. Gastric digestion of raw and roasted almonds in vivo. Journal of Food Science. 78:H1807-H1813.
- Liang, L., Chantry, C., Stephensen, C.B. 2010. Prevalence and risk factors For vitamin D deficiency among healthy infants in Sacramento, California. Pediatrics. 169:1337-1344.
- Hawkes, W.C., Alkan, Z. 2011. Delayed cell cycle progression from SEPW1 depletion is p53- and p21-dependent in MCF-7 breast cancer cells. Biochemical and Biophysical Research Communications. doi:10.1016/j.bbrc. 2011.08.032.
- Hawkes, W.C., Richter, D., Alkan, Z. 2013. Dietary selenium supplementation and whole blood gene expression in healthy North American men. Biological Trace Element Research. DOI:10.1007/s12011-013-9786-5.
- Turner, T., Burri, B.J., Haskell, M.J., Jamil, K.M., Jamil, M. 2013. Effect of daily consumption of �-cryptoxanthin-rich tangerines and �- carotene-rich sweet potatoes on vitamin A and carotenoid concentrations in plasma and breast milk of Bangladeshi women with low vitamin A status. American Journal of Clinical Nutrition. 98:1200-1218. DOI: 10.3945/ajcn. 113.058180.
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Progress 10/01/12 to 09/30/13
Outputs
Progress Report Objectives (from AD-416): Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti- inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. Objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. Objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/ obese individuals. Objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. Ojective 2B. Examine anti-inflammatory activities of polyphenol-rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. Objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of B-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. Objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. Objective 3B. Estimate the impact of daily consumption of food sources of CX or B-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. Objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. Objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. Objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Objective 5: Identify the role of dietary selenium and selenoproteins in regulating cellular responses to oxidative stress. Objective 5A. Identify the pro-inflammatory and anti-inflammatory proteins S-glutathionylated by selenoprotein W. Objective 5B. Determine the role of selenoprotein W in key inflammatory pathways. Approach (from AD-416): The impact of selenium, vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. The effect of selenium on cell division and cell signaling will be examined in cell culture. Replacing 5306-51530-013-00D (1/09). Objective 1: Recruitment of human volunteers for the study examining the effect of vitamin D supplementation on immune function was completed this year. Data analysis for the study is currently in progress. In addition, work with collaborating scientists continues to examine the effects of vitamin D supplementation on inflammation during pregnancy and during HIV infection. Objective 2A) Isolated lymphocytes were treated with physiological levels of resveratol glucuronide, the major metabolite of the plant antioxidant resveratrol, to define how this metabolite alters gene expression and function of these immune cells. Objective 2B) The U.S. High Bush Blueberry Council awarded a collaborative grant to study whether feeding humans blueberries will decrease the inflammatory response to a fatty meal. This study is complete and data analysis is in progress. Objective 2C) Mouse studies completed in 2010 by us showed that feeding purified polyphenols at levels higher than typically found in foods did not kill leukemia cells. The diet studies planned for this year with polyphenol-rich foods will not be pursued as this approach is likely to be ineffective. Objective 3: (1) We used in vitro methods for analyzing stomach digestion of carotenoids and retinoids from foods to determine the effects of added oil on carotenoid absorption from mandarin oranges. (2) We compared the effects of two food-based interventions, mandarin oranges and orange sweet potatoes, to positive and negative controls fed to Bangladeshi women with marginal vitamin A status. Beta-carotene concentrations increased in blood and breast milk of the group fed orange- fleshed sweet potatoes. Beta-cryptoxanthin concentrations increased in blood and breast milk of the group fed mandarin oranges. Vitamin A concentrations increased only in the blood and breast milk for the group fed vitamin A. Beta-cryptoxanthin was 4-fold more bioavailable than the beta-carotene and appeared in breast milk as well as serum. This study suggests that tangerines and perhaps similar fruits may be suitable for development as a food source for improving the vitamin A status. (3) We completed the chromatography analysis for tissue samples from our gerbil study. Major accumulation tissues were the liver and adipose. Carotenoid accumulation in most tissues increased with increasing dietary intake. Objective 4: The decision was made in a previous year that these studies would not be pursued due to insufficient funds. Objective 5: We discovered that selenoprotein W: (a) is required for the dimerization of the epidermal growth factor receptor; (b) mediates hydrogen peroxide-stimulated activation of the epidermal growth factor receptor and the hepatocyte growth factor receptor; (c) co-localizes with the epidermal growth factor receptor at the plasma membrane in starved cells; and, (d) is rapidly degraded after cells are stimulated with epidermal growth factor. Significant Activities that Support Special Target Populations: The work of this project on vitamin D supplementation directly benefits historically underserved segments of the population in that national survey data from the USDA shows that vitamin D deficiency is more common in African American and Mexican American segments of the US population. This work will provide a stronger scientific basis for dietary recommendations to these segments of the US population. Accomplishments 01 Improving vitamin D status in at-risk populations. Vitamin D deficiency is common in US youth with HIV infection and optimal vitamin D dosing for treatment is unclear, particularly because some antiretroviral drug regimens affect vitamin D metabolism. ARS scientists from the Davis, California, worked with colleagues from the Adolescent Trials Network to conduct a one year, placebo-controlled intervention trial of monthly dosing with 50,000 units of vitamin D3 among 203 HIV-infected youth ages 18-24. The use of the antiretroviral efavirenz was associated with lower baseline vitamin D status, but this supplementation regimen was safe and improved vitamin D status regardless of antiretroviral regimen. This study demonstrated that at- risk youth benefit from vitamin D supplementation levels comparable to those recommended for the general population. 02 Human lymphocyte function is affected by resveratrol glucuronide, a major in vivo metabolite of resveratrol. It is suspected that plant- derived antioxidants such as resveratrol, which is found in grapes and some berries, alter the activity of individual immune cells in the human body after consumption but specific mechanisms are not known. ARS scientists working at the Davis, California, location found that the major metabolite of resveratrol found in humans after ingestion, resveratrol glucuronide, altered gene expression in isolated T-helper lymphocytes, a type of white blood cell important in many immune responses. Such changes in gene expression suggest that changes in function also occurred. Further studies are in progress to define the mechanisms of action of this and other phytochemical metabolites generated in the body after consumption of a meal containing the parent compound. These results are novel and will help determine how plant antioxidants affect human health by modulating the activity of immune cells. 03 Bioavailability of provitamin A carotenoids from mandarin oranges and sweet potatoes. ARS scientists in Davis, California, working with colleagues in Bangladesh, compared the effects of two food-based interventions, mandarin oranges containing beta-cryptoxanthin and orange sweet potatoes containing beta-carotene, on carotenoid bioavailability and vitamin A status in Bangladeshi women with marginal vitamin A status. Beta-carotene concentrations increased in blood and breast milk of the group fed orange-fleshed sweet potatoes, while beta- cryptoxanthin concentrations increased in blood and breast milk of the group fed mandarin oranges. Vitamin A concentrations increased only in the blood and breast milk of the control group fed vitamin A capsules. Beta-cryptoxanthin was 4-fold more bioavailable than the beta-carotene and appeared in breast milk as well as serum. This study suggests that mandarin oranges may be suitable for development as a food source for improving vitamin A status. 04 Evaluation of biofortified cassava as a provitamin A-rich staple crop. Biofortified cassava, which was developed to combat vitamin A deficiency in Africa, also contains moderate amounts of beta-carotene. We estimated the effectiveness of biofortified cassava for increasing beta-carotene and vitamin A concentrations in ten healthy adult women. Data from this human study were combined with cassava consumption data from countries in Africa where cassava is a staple food, to model the daily vitamin A intake (and cyanide exposure, also contained in cassava) of women in Africa. If biofortified cassava completely replaced traditional low-carotenoid, white cassava in the diet, it could meet recommended vitamin A intakes for the following percentages of individuals from 6 African countries: Angola (95%), Central African Republic (95%), Congo (100%), Ghana (99%), Mozambique (99%), and Nigeria (92%). However, biofortified cassava loses substantial amounts of carotenoids during roasting when it is converted to gari, a traditional West African food made with cassava. Even so, this work demonstrates that consumption of biofortified cassava, processed to maintain beta-carotene and remove cyanide, can potentially increase vitamin A status in African populations and other areas of the world where cassava is a staple crop. 05 Selenoprotein W regulates hydrogen peroxide activation of receptor tyrosine kinases. Dietary selenium supplements are popular and are thought to protect against cancer, but the scientific evidence is contradictory. Other studies suggest selenium increases the incidence of type 2 diabetes (a major risk factor for hypertension, stroke and cardiovascular disease). Dysfunctions in receptor tyrosine kinases cause many cancers and receptor tyrosine kinases control the insulin and cytokine signaling that may be altered in type 2 diabetes. ARS scientists in Davis, California, found that the selenium-containing protein, selenoprotein W, regulates hydrogen peroxide-mediated activation of receptor tyrosine kinases. Thus, selenoprotein W�s regulation of receptor tyrosine kinase activation by hydrogen peroxide may provide insight into a possible common mechanism in the development of chronic disease, including cancer and type 2 diabetes.
Impacts
(N/A)
Publications
- Lafrano, M.R., Woodhouse, L.R., Burnett, D.J., Burri, B.J. 2013. Biofortified cassava increases �-carotene and vitamin A concentrations in the TAG-rich plasma layer of American women. British Journal of Nutrition. First View Article:pp 1-11. DOI: 10.1017/S0007114512005004.
- Katz, J.M., Lafrano, M.R., Winter, C.K., Burri, B.J. 2013. Modelling potential �-carotene intake and cyanide exposure from consumption of biofortified cassava. Journal of Nutritional Science. 2(e6):1-8. DOI:10. 1017/jns.2012.30.
- Stephensen, C.B. 2013. Provitamin A carotenoids and immune function. In: Tanumihardjo, S. Carotenoids and Human Health. New York, NY: Humana Press. pp. 261-270.
- Stephensen, C.B., Zunino, S.J. 2012. Nutrition and the immune system. In: Ross, C., Caballero, B., Cousins, R., Tucker, K., Ziegler, T., editors. Modern Nutrition in Health and Disease. 11th edition. Philadelphia, PA: Lippincott Williams & Wilkins, a division of Wolters Kluwer Health, Inc. p. 601-610.
- Burri, B.J. 2013. Carotenoids, chemistry, sources and physiology. In: Caballero, B., editor. Encyclopedia of Human Nutrition. 3rd Edition, Volume 1. Waltham, MA: Academic Press. p. 283-291.
- Zunino, S.J., Storms, D.H., Newman, J.W., Pedersen, T.L., Keen, C.L., Ducore, J.M. 2012. Dietary resveratrol does not delay engraftment, sensitize to vincristine, or inhibit growth of high-risk acute lymphoblastic leukemia cells in NOD/SCID mice. International Journal of Oncology. 41(6):2207-2212. DOI: 10.3892/ijo.2012.1650.
- Livingston, K.A., Jiang, X., Stephensen, C.B. 2013. CD4 T-helper cell cytokine phenotypes and antibody response following tetanus toxoid booster immunization. Journal of Immunological Methods. 390:18-29. DOI: 10.1016/j. jim.2013.01.001.
- Roman, M.J., Burri, B.J., Singh, R. 2012. Release and bioaccessibility of �-carotene from fortified almond butter preparations during in vitro digestion. Journal of Agricultural and Food Chemistry. 60(38).
- Zunino, S.J., Storms, D.H., Newman, J.W., Pedersen, T.L., Keen, C.L., Ducore, J.M. 2013. Oral or parenteral administration of curcumin does not prevent the growth of high-risk t(4;11) acute lymphoblastic leukemia cells engrafted into a NOD/SCID mouse model. International Journal of Oncology. 42:741-748. DOI: 10.3892/ijo.2012.1734.
- Turner, T., Burri, B.J. 2013. Potential nutritional benefits of current citrus consumption. Agriculture. 10:3390.
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti- inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. Objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. Objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/obese individuals. Objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. Ojective 2B. Examine anti-inflammatory activities of polyphenol-rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. Objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of B-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. Objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. Objective 3B. Estimate the impact of daily consumption of food sources of CX or B-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. Objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. Objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. Objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Objective 5: Identify the role of dietary selenium and selenoproteins in regulating cellular responses to oxidative stress. Objective 5A. Identify the pro-inflammatory and anti-inflammatory proteins S-glutathionylated by selenoprotein W. Objective 5B. Determine the role of selenoprotein W in key inflammatory pathways. Approach (from AD-416): The impact of selenium, vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. The effect of selenium on cell division and cell signaling will be examined in cell culture. Objective 1: A total of 18 subjects have now completed the full study. The study had been modified to examine the effect of vitamin D supplementation on vaccine-specific T-cell responses, in addition to the response of total T-cells. Thus subjects have been immunized with the tetanus toxoid vaccine as a booster immunization. Inclusion of this TT- specific endpoint has allowed reduction in the sample size to 24 subjects, based on data not available at the time the project was developed. Recruitment has stopped for the summer period because of improved vitamin D status at this time of year and to allow for laboratory analysis of samples that have been collected. Objective 2: (1) The U.S. High Bush Blueberry Council awarded a collaborative grant to study whether feeding humans blueberries will decrease the inflammatory response to a fatty meal. This study is in progress. (2) Feeding obese humans grapes decreased a type of LDL cholesterol linked to heart diseases, and increased the immune response against bacteria that is important in obese populations at high risk for developing infections. The mouse studies were not pursued due to insufficient resources. (3) Mouse studies completed in 2010 by us showed that feeding purified polyphenols at levels higher than typically found in foods did not kill leukemia cells. The diet studies planned for this year with polyphenol-rich foods will not be pursued as this approach is likely to be ineffective. Objective 3: We completed our food-based intervention study in Bangladesh, comparing the carotenoid and retinoid changes caused by feeding orange-fleshed sweet potatoes and mandarin oranges to lactating women with low vitamin A status. Beta-carotene concentrations in blood increased in the group fed orange-fleshed sweet potatoes and Beta- cryptoxanthin concentrations in blood increased in the group fed mandarin oranges. Beta-cryptoxanthin concentrations increased to a much greater extent than beta-carotene concentrations. This was similar to results we obtained in our in vitro digestion studies of beta-cryptoxanthin metabolism. We are now finishing analysis of the breast milk samples from the study. A second carotenoid intervention study using �biofortified� cassava varieties with good amounts of beta-carotene found increased beta- carotene and vitamin A concentrations in women. Therefore, eating properly prepared �biofortified� cassava has low risk, and can increase beta-carotene and vitamin A in people. Objective 5: We discovered a new mitogen-activated protein kinase pathway from MKK4 through p38-gamma, p38-delta, and JNK2 that controls phosphorylation of tumor suppressor protein p53 when selenoprotein W (SEPW1) is depleted. We showed SEPW1 is required for auto-phosphorylation and stabilization of the epidermal growth factor (EGF) receptor (EGFR). We demonstrated that activation of EGFR and 6 out of 6 other growth factor receptors present on breast cancer cells requires SEPW1. Significant Activities that Support Special Target Populations: Vitamin D insufficiency is common in African Americans due to low dietary intake and low dermal synthesis from sun exposure. Thus we are specifically recruiting subjects of African ancestry for this study as one of the entry criteria is vitamin D insufficiency. In addition, the results of this study will be of particular interest to African Americans as vitamin D deficiency is considered a health disparity condition affecting this community. Accomplishments 01 Biofortified cassava may prevent vitamin A deficiency. Cassava is an important staple crop in many tropical countries but the calories it provides are considered �empty� because it has a low content of vitamins and minerals. The goal of �biofortification� is to select natural varieties of staple foods, such as cassava, which have improved nutritional quality. Introduction of biofortified varieties of cassava that are naturally rich in pro-vitamin A carotenoids such as beta-carote could improve vitamin A intake in such settings, where vitamin A deficiency (which causes blindness and excess mortality) is often a significant public health problem. ARS scientists working at the Western Human Nutrition Research Center in Davis, CA, with a biofortified variet of cassava provided by the International Center for Tropical Agriculture in Cali, Colombia, evaluated the absorption of beta-carotene from this a standard cassava. The cassava varieties were fed to healthy women in a standard meal and the biofortified cassava produced a significantly greater increase in blood beta-carotene levels than the standard variety indicating good absorption and thus a likely beneficial effect on vitami A status. This study shows that biofortified cassava can be a good sourc of vitamin A and thus clears the way for larger studies to determine the effect of beta-carotene-rich cassava on preventing vitamin A deficiency countries where cassava is a staple crop. 02 Dietary vitamin D2 competes with vitamin D3. Vitamin D deficiency cause loss of bone mineral density and osteoporosis in adults and is common in the US because few foods contain either vitamin D2 (found in some plants and mushrooms) or vitamin D3 (of animal origin). Vitamin D3 is also produced in human skin by sun exposure but this source is limited by ski pigmentation (thus African Americans are at high risk of deficiency) and lack of sun exposure. ARS scientists from the Western Human Nutrition Research Center in Davis, CA, tested the availability of vitamin D2 from mushrooms treated with UV light to increase their vitamin D2 content. Th goal of the study was to determine if such mushrooms might be a useful source of vitamin D in the American diet. The study was done in healthy human volunteers during the summer and fall months and found that vitami D2 was readily absorbed from mushrooms (and a supplement provided as a control) but that vitamin D3 levels decreased proportionally resulting i no net improvement of vitamin D status. These findings indicate that increased intake of vitamin D2 will not improve vitamin D status for individuals with significant vitamin D3 �exposure� from other foods or from sun exposure, suggesting that vitamin D3 is the preferred form of vitamin D for dietary intake. 03 Selenium and cellular proliferation. Selenium is an essential mineral that is incorporated into a specific class of proteins, termed selenoproteins, with diverse and poorly understood cellular functions. Understanding the biology of such selenoproteins is necessary to understand how selenium affects human health, including cancer risk, whi may be decreased by the use of selenium supplements though data are equivocal on this point. ARS researchers working at the Western Human Nutrition Research Center in Davis, CA, discovered that selenoprotein W (SEPW1) is required for activation of a class of cellular proteins calle receptor tyrosine kinases, or RTKs, which are responsible for triggering cellular proliferation. Controlled cellular proliferation is normal and healthy but uncontrolled cellular proliferation is a hallmark of cancer. Since RTK dysfunction is known to be involved in the development of most cancers these findings suggest that selenium may affect cellular proliferation, and perhaps cancer risk, via its effects on the RTK proteins. More work is needed to further understand SEPW1 function, how selenoproteins might be targeted in cancer therapy, and to evaluate the potential benefits and risks of selenium supplementation. 04 Grapes are heart-healthy and boost immunity. Many studies show that adequate intake of fruits and vegetables are associated with low risk of heart disease and improvements in other measures of good health such as healthy immune system. However, data on the actual benefits of specific fruits and vegetables is quite limited. To address this deficit, ARS researchers working at the Western Human Nutrition Research Center in Davis, CA, investigated the effect of consuming table grapes (provided i a freeze-dried form for the sake of uniformity throughout the study peri on risk factors for heart disease, such as blood cholesterol, and marke of a healthy immune system, including the response of white blood cells stimulation with a product of bacteria. The study was conducted in obese adults and showed that eating grapes decreased blood levels of a class o cholesterol associated with heart disease, suggesting that consuming grapes may be heart-healthy. Consuming grapes also increased the respons of white blood cells to a microbial product, a finding consistent with improved protection against infections. These results suggest a benefici effect of consuming grapes in the obese population known to have a highe risk of both infections and heart disease.
Impacts
(N/A)
Publications
- Hawkes, W.C., Alkan, Z. 2012. Selenoprotein W depletion induces a p53- and p21-dependent delay in cell cycle progression in RWPE-1 prostate epithelial cells. Journal of Cellular Biochemistry. 113:61-69.
- Parelman, M.A., Storms, D.H., Kirschke, C.P., Huang, L., Zunino, S.J. 2012. Dietary strawberry powder reduces blood glucose concentrations in obese and lean C57BL/6 mice and selectively lowers plasma C-reactive protein in lean mice. British Journal of Nutrition. 1-11. DOI: 10. 1017/S0007114512000037.
- Zunino, S.J., Parelman, M., Freytag, T.L., Stephensen, C.B., Kelley, D.S., Mackey, B.E., Woodhouse, L.R., Bonnel, E. 2011. Effects of dietary strawberry powder on blood lipids and inflammatory markers in obese human subjects. British Journal of Nutrition. 1-10. DOI: 10. 1017/S0007114511006027.
- Turner, T., Burri, B.J. 2012. Rapid isocratic HPLC method and sample extraction procedures for measuring carotenoid, retinoid, and tocopherol concentrations in human blood and breast milk for intervention studies. Chromatographia. DOI 10.1007/s10337-012-2193-9.
- Havens, P.L., Stephensen, C.B., Hazra, R., Flynn, P.M., Wilson, C.M., Rutledge, B., Bethel, J., Pan, C.G., Woodhouse, L.R., Van Loan, M.D. 2012. Vitamin D3 decreases parathyroid hormone in HIV-infected youth being treated with tenofovir: a randomized, placebo-controlled trial. Clinical Infectious Diseases. 54(7):1013-1025. DOI: 10.1093/cid/cir968.
- Turner, T., Burri, B.J., La Frano, M.R. 2012. Beta-cryptoxanthin: A vitamin A-forming carotenoid. In: Yamaguchi, M., editor. Carotenoids: Properties, Effects and Diseases. Hauppauge, NY: Nova Publishers. p. 331- 354.
- Zunino, S.J., Storms, D.H., Newman, J.W., Pedersen, T.L., Keen, C.L., Ducore, J.M. 2012. Resveratrol given intraperitoneally does not inhibit growth of high-risk t(4;11) acute lymphoblastic leukemia cells in NOD/SCID mouse model. International Journal of Oncology. 40(4):1277-84. DOI:10. 3892/IJO.2011.1316.
- Burri, B.J. 2012. Evaluating global barriers to the use of red palm oil as an intervention food to prevent vitamin A deficiency. Journal of Food Science. 11:221-233. DOI: 10.1111/j.1541-4337.2011.00181.x.
- Stephensen, C.B., Zerofsky, M., Burnett, D., Lin, Y., Hammock, B.D., Hall, L.M., Mchugh, T.H. 2012. Ergocalciferol from mushrooms or supplements consumed with a standard meal increases 25-hydroxyergocalciferol but decreases 25-hydroxycholecalciferol in the serum of healthy adults. Journal of Nutrition. 142(7):1246-1252. DOI: 10.3945/jn.112.159764.
- Hawkes, W.C., Alkan, Z. 2012. Delayed cell cycle progression in selenoprotein W depleted cells is regulated by a mitogen-activated protein kinase kinase 4�p38�p53 pathway. Journal of Biological Chemistry. DOI: 10. 1074/jbc.M112.346593.
- Armstrong, P., Kelley, D.S., Newman, J.W., Staggers, F., Hartiala, J., Allayee, H., Stephensen, C.B. 2012. Arachidonate 5-lipoxygenase gene variants affect response to fish oil supplementation by healthy African Americans. Journal of Nutrition. 142(8):1417-1428. DOI: 10.3945/jn.112. 159814.
- Stephensen, C.B. 2012. Vitamin A and immune function. In: Preedy, V.R., editor. Food and Nutritional Components in Focus No. 1: Vitamin A and Carotenoids: Chemistry, Analysis, Function and Effects. Cambridge, UK: Royal Society of Chemistry. p. 501-515. DOI:10.1039/ISBN
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti- inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. Objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. Objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/obese individuals. Objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. Ojective 2B. Examine anti-inflammatory activities of polyphenol-rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. Objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of B-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. Objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. Objective 3B. Estimate the impact of daily consumption of food sources of CX or B-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. Objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. Objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. Objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Objective 5: Identify the role of dietary selenium and selenoproteins in regulating cellular responses to oxidative stress. Objective 5A. Identify the pro-inflammatory and anti-inflammatory proteins S-glutathionylated by selenoprotein W. Objective 5B. Determine the role of selenoprotein W in key inflammatory pathways. Approach (from AD-416) The impact of selenium, vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. The effect of selenium on cell division and cell signaling will be examined in cell culture. Objective 1: A pilot study of eight subjects to establish methods for measuring the production of cytokines by vaccine-specific T cells was completed. We began recruitment of subjects with vitamin D insufficiency for the main study. By the end of July, 188 subjects had been screened for enrollment by telephone, 21 had come for screening visits, ten had enrolled and six had completed the study. Enrollment is slower than planned and we have thus expanded recruitment from Davis to Sacramento and are now specifically recruiting African Americans due to their greater risk of vitamin D insufficiency. Objective 2: (1) Analysis of data from our previously completed human trial showed that consuming strawberries increased the number of cytotoxic T cells and the production of the cytokine tumor necrosis factor, both being important for immune responses to bacteria and viruses. A second study is in progress to evaluate the effect of grapes on inflammation in people at risk for cardiovascular disease. (2) The High Bush Blueberry Council awarded a grant to determine if blueberries will decrease the inflammatory response to a fatty meal. Data analysis from earlier studies with a mouse model for diet-induced obesity and diabetes found that feeding strawberries reduced blood glucose and serum C- reactive protein. (3) Data analysis from earlier mouse studies found that feeding purified polyphenols at levels higher than found in foods did not kill leukemia cells. Thus the diet studies planned for this year with polyphenol-rich foods will not be pursued as this approach is likely to be ineffective. Objective 3: (1) In vitro methods were developed to analyze carotenoid and retinoid absorption from foods, including sweet potatoes, cassava and mandarin oranges. The methods are being modified for use with an �artificial stomach� model to better predict absorption. (2) Enrollment for the beta-cryptoxanthin study is complete. Laboratory and data analysis is in progress. Baseline characteristics did not differ between groups, showing that randomization was effective. (3) New extraction and chromatography methods were developed for small amounts of tissues, and tissue samples are being analyzed from the gerbil study. Objective 5: We developed a method to measure selenoprotein W1 in cell culture to determine if silencing its transcription also decreases protein levels, which was confirmed. We then explored how this silencing causes cell cycle arrest, a phenomenon that we reported previously in epithelial cells whose growth is stimulated by epidermal growth factor. Briefly, silencing promotes a cascade of events including activation of the enzyme p38 to add phosphate residues to the regulatory protein p53 on serine 33 and 46. This blocks p53 degradation and allows its accumulation in the nucleus where it induces p21 gene transcription. The protein p21 then induces arrest. We also found that selenoprotein W1 was associated with the epidermal growth factor receptor complex, and that cell cycle arrest was not due to loss of antioxidant activity or DNA damage. Significant Activities that Support Special Target Populations The vitamin D research described under the first accomplishment has particular importance for African Americans because of their greater risk of vitamin D deficiency. Accomplishments 01 Strawberries and resistance to infectious diseases. Understanding how specific foods promote good health will improve the quality of dietary recommendations. ARS scientists at the Western Human Nutrition Research Center in Davis, California, determined that feeding strawberries to obe individuals increased their cytotoxic T cell response and the production of tumor necrosis factor alpha by monocytes. Cytotoxic T cells and monocytes are important for preventing bacterial and viral infections. Obese individuals are at higher risk for infections than normal weight people. Eating strawberries may be a means of balancing important immun responses in individuals who are at risk for developing infections. 02 Strawberries and type 2 diabetes. Loss of control of blood sugar (gluco , as occurs in type 2 diabetes, leads to adverse health consequences whi might be ameliorated by improving diet quality. ARS scientists at the Western Human Nutrition Research Center in Davis, California, determined that feeding a diet containing strawberry powder (prepared from whole freeze-dried strawberries) reduced blood glucose levels in both lean and obese mice, and reduced C-reactive protein in lean mice. A mouse model for diet-induced obesity and diabetes was used for these studies to represent humans consuming high dietary fat. Eating strawberries may th be beneficial for controlling blood sugar in obese humans with type 2 diabetes 03 Selenoprotein W1 and the cell cycle. Selenium deficiency causes immune deficiency and other adverse health outcomes but the role of specific selenoproteins in mediating such effects is largely undefined. ARS scientists at the Western Human Nutrition Research Center in Davis, California, have examined the role of selenoprotein W1 in a basic cellul process, signaling from a cell-surface receptor, the epidermal growth factor receptor, to regulate cellular proliferation. The scientists fou that selenoprotein W1 is required for epidermal growth factor signaling and thus for cellular proliferation, and that selenoprotein W1 is associated with three proteins that are components of the activated epidermal growth factor receptor complex. Because selenoprotein W1 is decreased by low dietary intake of selenium this mechanism may explain, least in part, the relationship between low dietary selenium intake and decreased immune function. 04 Methods to study the effect of diet on vaccine responses. Vaccine- specific T cells are rare in peripheral blood and it is thus difficult t measure the effect of nutritional interventions on aspects of T cell- mediated immunity that have been well-studied in animal model systems. address this problem, ARS scientists at the Western Human Nutrition Research Center in Davis, California, have tested a method to expand and subsequently identify cytokine production from vaccine-specific T cells from peripheral blood, following one week of amplification in cell cultu Using this method in healthy adults the scientists found a broader tha expected pattern of cytokine production by tetanus vaccine-specific T cells, including cytokines from the major groups of T-helper (Th) cells, including Th1, Th2, Th17 and Treg cells. These methods can now be used nutritional intervention studies to characterize the effect of diet on immune function and resistance to infection or autoimmune disease. 05 Availability of vitamin A-forming carotenoids from foods. The amount of vitamin A that carotenoid containing foods provide depends not only on t amounts of the carotenoid in the food, but also on how well the carotenoids are absorbed. ARS scientists at the Western Human Nutrition Research Center in Davis, California, used in-vitro digestion methods to compare the apparent bioavailabilities of mandarin oranges and sweet potatoes, excellent dietary sources of the vitamin A-forming carotenoids beta-carotene, and beta-cryptoxanthin, respectively. Beta-cryptoxanthin from mandarin oranges was approximately 240% more bioavailable than beta carotene from sweet potatoes. This result supports earlier research showing that eating comparable amounts of beta-cryptoxanthin and beta- carotene-rich foods resulted in much higher beta-cryptoxanthin concentrations in the blood. This suggests that beta-cryptoxanthin-rich foods, which include citrus fruit, might be better sources of vitamin A than has traditionally been expected based on their relatively low abundance in the diet.
Impacts
(N/A)
Publications
- Burri, B.J. 2011. Evaluating the sweet potato as an intervention food to prevent vitamin A deficiency. Comprehensive Reviews in Food Science and Food Safety. 10:118-130.
- Makhoul, Z., Kristal, A.R., Gulati, R., Luick, B., Bersamin, A., O'Brien, D., Hopkins, S.E., Stephensen, C.B., Stanhope, K.L., Havel, P.J., Boyer, B. 2011. Associations of obesity with triglycerides and C-reactive protein are attenuated in adults with high red blood cell eicosapentaenoic and docosahexaenoic acids. European Journal of Clinical Nutrition. 65(7):808- 817. Available:
- Stephensen, C.B., Armstrong, P., Newman, J.W., Pedersen, T.L., Legault, J., Schuster, G., Kelley, D.S., Vikman, S., Hartiala, J., Hooman, A. 2011. ALOX5 gene variants affect eicosanoid production and response to fish oil supplementation. Journal of Lipid Research. 52(5):991-1003.
- Hartiala, J., Li, D., Conti, D.V., Vikman, S., Patel, Y., Tang, W., Brennan, M., Newman, J.W., Stephensen, C.B., Armstrong, P., Hazen, S.L., Allayee, H. 2011. Genetic contribution of the leukotriene pathway to coronary artery disease. Human Genetics. 129(6):617-627.
- Sharkar, M., Jou, M., Hossain, M.B., Lonnerdal, B., Stephensen, C.B., Raquib, R. 2011. Prenatal zinc supplementation of zinc-adequate rats adversely affects immunity in offspring. Journal of Nutrition. 141(8):1559- 1564.
- Burri, B.J., Chang, J.S., Turner, T. 2011. Citrus can help prevent vitamin A deficiency in developing countries. California Agriculture. 65(3):130- 135.
- Burri, B.J., Chang, J., Neidlinger, T.R. 2011. Beta-Cryptoxanthin- and alpha-carotene-rich foods have greater apparent bioavailability than beta- carotene-rich foods in Western diets. British Journal of Nutrition. 105:212-219.
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti- inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. Objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. Objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/obese individuals. Objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. Ojective 2B. Examine anti-inflammatory activities of polyphenol-rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. Objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of B-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. Objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. Objective 3B. Estimate the impact of daily consumption of food sources of CX or B-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. Objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. Objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. Objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Objective 5: Identify the role of dietary selenium and selenoproteins in regulating cellular responses to oxidative stress. Objective 5A. Identify the pro-inflammatory and anti-inflammatory proteins S-glutathionylated by selenoprotein W. Objective 5B. Determine the role of selenoprotein W in key inflammatory pathways. Approach (from AD-416) The impact of selenium, vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. The effect of selenium on cell division and cell signaling will be examined in cell culture. Replacing 5306-51530-013-00D (1/09). Objective 1: (1) A pilot study was completed involving six subjects to evaluate newly implemented methods for testing antigen-specific T-cell responses that should be influenced by vitamin D status. This involved implementing new methods in our laboratory. (2) Donation of vitamin D supplements was arranged and the supplements analyzed to assure appropriate content. The intervention will begin before December, 2010. Objective 2: (1) The California Strawberry Commission funded a human study to determine the anti-inflammatory effects of strawberries in obese individuals. The data show dietary strawberries inhibited T cell proliferation, and beneficially altered several serum lipid fractions. California Table Grape Commission funded a human study to understand the anti-inflammatory role of dietary grape powder in obese human volunteers. (2) A mouse model was used to determine whether dietary strawberry powder inhibited inflammation associated with diet-induced obesity and measurements of inflammation are being completed. (3) Mice engrafted human leukemia cells were fed or injected with resveratol and curcumin to determine if these phytochemicals could slow the progression of the leukemia. Neither treatment was effective. Objective 3: (1) 3-hydroxyretinol and didehydroretinol, major metabolites of beta-cryptoxanthin were synthesized. A rapid, high- performance liquid chromatography method was developed for measuring beta- cryptoxanthin and its metabolites. In vitro methods for analyzing stomach digestion of carotenoids and other nutrients from foods were tested. Analysis of metabolites in human blood and tissues has begun. (2) Standard operating procedures for dietary and clinical analysis were developed, and a collaborative study to assess the impact of daily consumption of food sources of beta-cryptoxanthin and beta-carotene on vitamin A status was begun. (3) Methods to determine the impact of beta- cryptoxanthin on immune and bone marker status in the Mongolian gerbil are being selected and optimized for our laboratory conditions. Objective 4: These objectives have not been pursued because funds are not available. Extramural funds are required to pursue these objectives. Objective 5: We tried three different methods to show that selenoprotein W (SEPW1) affected global protein glutathionylation and none of them worked. Immunoprecipitation of p53 failed to reveal glutathionylation of this protein. We ruled out ER stress and the unfolded protein response as mediators of cell cycle arrest induced by SEPW1 depletion. We determined that p53 and p21 levels are not different in RWPE-1 cell lines stably under- or over-expressing SEPW1. We showed that SEPW1 knockdown induced a G1 arrest in MCF-7 breast cancer cells. We showed that G1 arrest from SEPW1 knockdown is abolished in MCF-7 mutant cells in which p53 expression is silenced and in RWPE-1 and MCF-7 cells in which p21 expression was silenced. We showed that SEPW1 knockdown causes p53 to be phosphorylated on serine 33, consistent with involvement of the p38 stress activated MAP kinase, glycogen synthase kinase beta, cyclin-dependent kinase 7 and/or 9. Significant Activities that Support Special Target Populations The research described under the first accomplishment has particular importance for African Americans because of the high risk of heart disease in that group and because of the high prevalence in African Americans of the ALOX5 gene variant examined in that study, which is associated with a high risk of heart disease. Accomplishments 01 Fish oil for African Americans to decrease risk of heart disease: Both diet and genes contribute to one�s risk of heart disease and in the case of omega-3 fatty acids and the ALOX5 gene the two factors may interact. ARS researchers at Davis, CA found that African Americans with a variant of the ALOX5 gene that is associated with a high risk of heart disease produce lower levels of potentially heart-healthy omega-3 metabolites th did subjects with the low-risk ALOX5 variant. Subjects with the high-ris variant also responded to fish oil supplements with a smaller increase i these metabolites than did subjects with the low-risk variant. These da suggest that dietary recommendations for lowering risk of heart disease may need to be tailored by genotype to maximize their impact. 02 Strawberries decrease cardiovascular risk: The risk of heart disease is higher among obese than non-obese Americans but this risk can be decreas by dietary interventions. ARS reseachers at Davis, CA demonstrated that freeze-dried strawberry powder consumed by obese individuals decreased serum cholesterol and improved other serum lipid indicators that are linked to risk of heart disease and stroke. These results indicate that consuming strawberries should decrease the risk of heart disease by improving lipid metabolism and will thus help form the scientific basis dietary guidelines for all Americans. 03 Carotenoid bioavailability and food-based interventions: Studies show lo correlations between dietary intake and serum concentrations of lycopene and other carotenoids, indicating that serum concentrations alone are a poor biomarker for assessing the effectiveness of dietary interventions with these phytonutrients. ARS researchers at Davis, CA showed that combining dietary intake methods (food-frequency questionnaires and 3-d diet records) using the triads method improved the validity of this relationship. Furthermore, correcting dietary information for differenc in lycopene absorption from its most important food matrices further improved validity, to a relatively strong level. Thus, using the triads method with bioavailability estimates should make it possible to determi whether food-based interventions for increasing lycopene or other carotenoids are effective. 04 Selenium and the cell cycle: Although selenium supplements prevent canc in animals, human cancer-prevention trials have failed, perhaps because do not understand what most of the 25 selenium-containing proteins (�selenoproteins�) do in the body. Selenoprotein W (SEPW1) is the most ancient and widely distributed selenoprotein in Nature but its function unknown. ARS researchers at Davis, CA have discovered that SEPW1 control the stability of p53, a tumor suppressor protein that plays a central ro in controlling the cell-division cycle, which is required for all cells replicate themselves and is thus fundamental to growth and development f all organisms. This new knowledge adds to our understanding of selenium biology and will help form a solid scientific basis for dietary recommendations for Americans with regard to maintaining optimal health. 05 Resveratrol and curcumin do not improve leukemia outcome: Plant polyphenols and other dietary components may slow the development or progression of certain cancers but results vary by cancer type. ARS researchers at Davis, CA found that feeding resveratrol and curcumin to leukemic mice was not effective in decreasing the growth of leukemia cel nor were these polyphenols able to increase the effectiveness of vincristine, a standard chemotherapeutic agent. These results do not support a role for these compounds for slowing progression of establishe leukemia and thus help form a better scientific basis for dietary recommendations to individuals with this form of cancer.
Impacts
(N/A)
Publications
- Ahmad, S.M., Haskell, M.J., Raqib, R., Stephensen, C.B. 2009. Vitamin A Stores Are Associated With T-Cell Responses In Bangladeshi Men. British Journal of Nutrition. 102:797-802.
- Hawkes, W.C., Alkan, Z. 2010. Regulation of Redox Signaling by Selenoproteins. Biological Trace Element Research. 134(3): 235-251, 2010.
- Zunino, S.J., Storms, D.H., Ducore, J. 2010. Novel in vivo model of inducible multidrug resistance in acute lymphoblastic leukemia with chromosomal translocation t(4;11). Cancer Letters. 296:49-54.
- Hawkes, W.C., Wang, T.T., Alkan, Z., Richter, D., Dawson, K. 2009. Selenoprotein W Modulates Control of Cell Cycle Entry. Humana Press Inc, Biological Trace Element Research. 131:229-244.
- Hawkes, W.C., Hwang, A., and Alkan, Z. 2009. The Effect of Selenium Supplementation on DTH Skin Responses in Healthy North American Men. Journal of Trace Elements in Medicine and Biology. Vol.23:272-280.
- Boyer, B.B., Mohatt, G.V., Plaetke, R., Herron, J., Stanhope, K.L., Stephensen, C.B., Havel, P.J. 2007. Metabolic Syndrome in Yup'ik Eskimos: The Center for Alaska Native Health Research (CANHR) Study. Obesity. Vol. 15 No.11, 2535-2540.
- Eneroth, H., Arifeen, S., Persson, L., Lonnerdal, B., Hossain, M.B., Stephensen, C.B., Ekstrom, E. 2010. Maternal Multiple Micronutrient Supplementation Has Limited Impact on Micronutrient Status of Bangladeshi Infants Compared with Standard Iron Folic Acid Supplementation. Journal of Nutrition. doi:10.3945/jn.109.111740.
- Humphrey, B.D., Stephensen, C.B., Calvert, C.C., Klasing, K.C. 2006. Lysine deficiency and feed restrition independently alter cationic amino acid transporter expression in chickens. Comparative Biochemistry and Physiology. Part A 143:218-227.
- Humphrey, B.D., Stephensen, C.B., Calvert, C.C., Klasing, K.C. 2004. GLUCOSE AND CATIONIC AMINO ACID TRANSPORTER EXPRESSION IN GROWING CHICKENS (GALLUS GALLUS DOMESTICUS). Comparative Biochemistry and Physiology. 138:515-525.
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) Objective 1: Conduct a controlled, vitamin D supplementation trial in volunteers with vitamin D insufficiency (VDI) to determine if supplementation to achieve the proposed level of >75 nmol/L for maintenance of bone health is also appropriate for maintenance of immune function. � Sub-objective 1A. Determine if supplements decrease the production of proinflammatory and increase the production of anti-inflammatory cytokines and chemokines by innate immune cells stimulated ex vivo. � Sub-objective 1B. Determine if supplements decrease serum markers of inflammation and autoimmune activity, and increase serum levels of defensive molecules. � Sub-objective 1C. Determine if supplements decrease blood levels of proinflammatory T-helper type 1 (Th1) and Th17 cells and increase levels of anti-inflammatory T-regulatory (Treg) and Th2 cells. Objective 2: Determine the impact of plant polyphenols and polyphenol- rich foods on immune cell function using cell culture systems, mouse models, and human volunteers. Examine anti-inflammatory and anti-cancer activities of polyphenols in animal models, as well as inflammation and oxidative damage in studies with human volunteers, including overweight/obese individuals. � Sub-objective 2A. Analyze the effects of polyphenol-rich foods and individual plant polyphenols on immune cell function in vivo and ex vivo. � Sub-objective 2B. Examine anti-inflammatory activities of polyphenol- rich foods, individual plant polyphenols and vitamin A in mice and humans who are at risk for developing inflammatory disease, such as autoimmune mice and obese humans. � Sub-objective 2C. Evaluate the anti-cancer activity of polyphenol-rich foods and individual plant polyphenols. Objective 3: Examine the absorption of '-cryptoxanthin (CX) from supplements and foods, its contribution to vitamin A stores, and the impact of CX, other carotenoids and vitamin A on immune function. � Sub-objective 3A. Measure the absorption and metabolism of CX from Satsuma mandarin juice fed to healthy adult humans. � Sub-objective 3B. Estimate the impact of daily consumption of food sources of CX or '-carotene (BC) on plasma and breast milk concentrations of CX, BC and retinol in lactating women. � Sub-objective 3C. Determine the impact of CX on immune and bone marker status in the Mongolian gerbil. Objective 4: Determine if high-level vitamin A intake is associated with higher Th2 and Treg responses and lower Th1 and Th17 responses relative to adequate and deficient intake. � Sub-objective 4A. Using dietary and targeted gene disruption approaches in mice, determine if vitamin A enhances Th2 and Treg responses by acting directly on T cells. � Sub-objective 4B. Using subjects recruited in the vitamin D supplementation trial described under Objective 1, determine if vitamin A status is associated with higher blood levels of NK, NK-T, Th2 and Treg cells, and lower levels of Th1 and Th17 cells. Approach (from AD-416) The impact of vitamins A and D, and plant polyphenols, on immune function will be examined using cell culture systems, mouse models, and human intervention trials. The anti-cancer activities of polyphenols will be examined in animal models. Absorption of beta-cryptoxanthin will be examined in gerbils and humans. Replacing 5306-51530-013-00D (1/09). Significant Activities that Support Special Target Populations Subobjectives 1A-C: We have submitted an application to the human subjects review committee and are implementing new lab methods to be used for the pilot phase of this study. Subobjective 2A: The California Table Grape Commission awarded a grant for a human study to understand the anti- inflammatory role of grape powder in human volunteers at risk for developing cardiovascular disease. The agreement has been signed and transfer of funds to the USDA is in progress. Subobjective 2B: This human study to examine the anti-inflammatory effects of strawberries in obese individuals has been completed. Laboratory work and data analysis are in progress. Subobjective 2C: The signal transduction pathways for the phytochemical parthenolide (from the herb feverfew) has been analyzed in leukemia cells to determine the mechanism by which parthenolide induces cell death. Immunodeficient mice were used to study whether resveratrol or curcumin can kill leukemia. The leukemia was allowed to grow in immunodeficient NOD/SCID mice and resveratrol or curcumin was injected intraperitoneally every other day until the mice succumbed to the disease. Survival curves were compared between mice receiving the resveratrol, curcumin, the chemotherapeutic agent vincristine, or control solution. Lab work and data analysis are in progress. Subobjective 3A: Funds are not available for human study. A study on CX metabolism and vitamin A formation in gerbils was conducted and data is being analyzed that will lead to information that will decrease time and costs of the human study, possibly making it feasible. Subobjective 3B: Progress is described under number 7, International Cooperation. Subobjective 3C: Results from gerbil metabolism study are being analyzed and will be used to plan gerbil intervention study on immune response and bone metabolism. Subobjective 4A: This aim is not currently being pursued due to insufficient funds. Producing transgenic mice from the Rara targeting construct we have produced will require extramural funds, for which we have submitted an extramural grant proposal. Subobjective 4B: We have applied for extramural funds to pursue this aim using a revised approach involving comparison of continuous low-dose vitamin A supplements vs. high-dose supplements to increase liver stores that will not be feasible with subjects recruited under Subobjective 1.
Impacts
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Publications
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