Brief Summary of Minutes of Annual Meeting

Meeting was called to order May 29, 2014 at 8:00 a.m. EST. Welcome and Introductions: Participants were welcomed by the host and Meeting Chair, Dr. Connie Weaver, Purdue University, Nutrition Science; Dr. Karen Plaut, Senior Associate Dean for Research & Faculty Affairs, College of Agriculture, Purdue University; and Dr. Tammy Bray, Dean, College of Public Health and Human Sciences, Oregon State University (Administrative Advisor). Meeting objectives, timeline, and responsibilities were discussed. Dr. David Sands was elected by the members present to be the chair of the next annual meeting. He will coordinate next year’s W3002 meeting in Bozeman, Montana; targeted for the first week in August, 2015. Juan Andrade, University of Illinois was selected to be secretary. Dr. Deirdra Chester, NIFA Program Leader, joined the meeting via Skype and provided an update on the NIFA budget and current and future funding opportunities. Each W3002 Investigator attending the meeting provided an oral progress report in the following order: 1) 'A tale of three stores: holocarboxylase synthetase gene repression complexes, milk-borne microRNAs, and mitochondrial acetyl-CaA carboxylase 2,' Janos Zempleni, University of Nebraska, Lincoln 2) 'Micronutrient deficiencies in Ethiopia,' Barbara Stoecker, Oklahoma State University 3) 'Green tea and NASH,' Rich Bruno, Ohio State University 4) 'Diet, epigenetics and chronic disease,' Emily Ho, Oregon State University 5) 'Low-cost technologies to support development efforts in nutrition and health,' Juan Andrade, University of Illinois 6) 'Investigating the nutrition quality of new fortified-blended foods,' Brian Lindshield, Kansas State University 7) 'Influence of nanoemulsions for nutrient bioavailability,' Yeonhwa Park, University of Massachusetts 8) 'Paying more attention to breeding plants for nutrition: The Breeder’s Dilemma,' David Sands, Montana State University 9) 'Botanicals and Bone,' Connie Weaver, Purdue University 10) 'Effect of sleep deprivation on body weight and energy expenditure,' Jennifer Teske, University of Arizona 11) 'Update on purple potatoes,' Jairam Vanamala, Pennsylvania State University Discussion Zempleni – Vanamala: Processing effects on miRNA Zempleni – Weaver: Bone effects of ACC1 Andrade – Lindshield: Iron indicators Stoecker – Ho: Zinc biomarkers Bruno – Ho: Grant to AICR Sands – Lindshield: Incorporation of high lysine into nutritional supplement Sands – Andrade: Analyze minerals in cereals Teske – Park: CLA feeding and energy expenditure Invitation to test in model systems purple potatoes (Vanamala) and blueberries (Weaver) The meeting adjourned at 3:00 p.m. May 30, 2014: The following invited speakers from Purdue University gave brief reports on related research to W3002: 1) 'Phosphorus bioavailability,' Kathleen Hill Gallant 2) 'Upregulation of our Nrf2 by phytochemicals and impact on diabetes,' John Burgess 3) 'Bioavailability and bioactivity of vitamin E metabolites,' Qing Jiang

The participants of the multi-state project have been highly productive during the past reporting period as evidenced by 102 peer-reviewed publications among participants and enhancement of collaborations between project members. Project objectives are listed below along with scholarship activities of the lead station W3002 OBJECTIVES: 1. Determine the bioavailability (absorption, distribution, metabolism, elimination) of nutrients and other food components. 2. Evaluate the bioactivity of nutrients and other food components in order to elucidate their underlying protective mechanisms. ACCOMPLISHMENTS University of Arizona (Jennifer A. Teske): Insufficient sleep may alter nutrient bioavailability through changes in the gut-brain bidirectional axis, which is integral to gastrointestinal functions. We use validated rodent models of sleep deprivation (exposure to pre-recorded environmental noise in rats models human weight gain and hyperphagia and a murine model of sleep deprivation that models continued interruption in hospitalized patients) to study the link between health and sleep. We focus on the role of orexin, an endogenous neuropeptide critical for sleep/wake and body weight regulation. Our data suggest sleep deprivation reduces orexin signaling in key sleep regulatory brain sites to in turn cause weight gain. University of California, Berkeley (Barry Shane): We have continued studies on the metabolic and nutritional effects of common polymorphisms in human folate-related genes that have been shown to influence disease risk. We have continued to evaluate the B12-dependent methionine synthase (MS) and methylene-tetrahydrofolate reductase (MTHFR) genetic mouse models to mimic the effects of these polymorphisms and to evaluate their effects on metabolism and how this is modified by nutritional status. We have developed a mouse model that mimics the clinical effects of human B12 and folate deficiency, and which will allow us to investigate potential adverse effects of high folate intake. We continue to evaluate genetic risk factors for neural tube defects and to identify putative modifier genes which influence folate status, homocysteine levels, and methylation potential using a number of mouse strains and a cohort of students at Trinity College, Dublin. University of Connecticut (Ji-Young Lee): With the increasing obesity epidemic, co-morbidities of obesity, notably insulin resistance, type 2 diabetes, cardiovascular disease, and nonalcoholic fatty liver disease, are major health problems in the U.S. Understanding of the mechanisms underlying the bioactivities of food components is prerequisite to the successful development of preventive strategies against obesity and obesity-associated disorders. It is widely accepted that chronic inflammation due to increased macrophage infiltration in the lipid-laden adipose tissue increases the release of excess free fatty acids that are taken up by other tissues such as skeletal muscle, ultimately disrupting insulin signaling. Blackcurrant (Ribes nigrum) is among the berries with high polyphenol contents. Since blackcurrant farming started in the U.S. in 2003, it has gained popularity due to its potential health benefits. Our preliminary studies suggest that blackcurrant prevents adipose inflammation and hyperglycemia, enhances the expression of genes involved in mitochondrial biogenesis/bioenergetics, and decreases the inflammatory action of splenic monocytes in diet-induced obesity mice. OUTCOMES: The results from this project were disseminated by oral or poster presentation at the Experimental Biology in April 2014 and World Congress on the Role of Inflammation in Exercise hosted by the American College for Sports Medicine in May 2014. University of Illinois, Urbana-Champaign (Juan Andrade): Our research focuses on using low-cost technologies or approaches to improve nutrition in developing countries. Lately, the following research themes have been pursued: 1) development of an immunological base photonic crystal biosensor for the assessment of iron deficiency anemia; 2) development of an extruded product for delivery of iron into nixtamalized corn masa; and 3) use of an International Service and Experiential Learning (ISEL) program to bridge outreach, education and research for enhanced internationalization of undergraduate students at Illinois. OUTCOMES: 1. Biosensor (knowledge). The development of diagnostic technologies using photonic crystal biosensors was improved by the use of iron oxide nanoparticles as conjugated labels. These nanoparticles allow for increased sensitivity and potential separation functionality for biomarker molecules in complex fluids. 2. Fortification (knowledge). This is the first application of a superfortified extruded puff material as a delivery vehicle of nutrients into a wet matrix (nixtamalized corn) without changing sensory characteristics of final masa (dough) for tortillas; a staple commonly consumed throughout Central America. 3. Education (knowledge). The ISEL program enabled students to be more knowledgeable on global issues linking agriculture, nutrition and health. Abroad programs merging service, learning and outreach can support students’ attitudes towards service, reduced their ethnocentrism and promote their communication abilities with others by reducing their apprehension to engage in discussion with peers from other countries. Iowa State University (Wendy White): We completed two major human metabolic studies funded by Unilever R&D, Vlaardingen, The Netherlands. We investigated the dose-response relation between the amount of soybean oil in salad dressing and the extent of absorption of eight fat-soluble nutrients/bioactives from salad vegetables, including carotenoids, tocopherols, and phylloquinone. We also quantified the absorption of retinyl palmitate formed from the provitamin A carotenoids in the salad vegetables. A second study investigated the effects of adding lecithin to the soybean oil in salad dressing on the bioavailability of the same fat-soluble nutrients/bioactive food components in the salad vegetables. Kansas State University (Brian Lindshield): A micronutrient-fortified corn-soybean blend is the most commonly used food aid product. However, given climate change there are questions about whether these crops are sustainable choices for food aid. Sorghum and cowpeas are drought tolerant crops that are also not genetically modified. The latter is a preference of some countries where food aid is provided. Our objective is to determine whether extruded sorghum-cowpea blends can provide equal, or better, nutritive value than traditional corn-soybean blend. The effectiveness of blends will be determined in a field trial of Tanzanian children under the age of 5 at risk of micronutrient deficiency. University of Massachusetts (Yeonhwa Park): Nanoemulsions are being used to design effective delivery systems for lipophilic bioactive food components. These various delivery systems can contain different functional food ingredients without adversely affecting their desirable properties. We have tested bioavailability of Coenzyme Q10 and other fatty acids on various delivery systems. Results with in vitro digestion models have been correlated to rodent feeding trials. Montana State University (David Sands): Camelina sativa is a new omega-3 oil and protein crop that in many ways rivals soy. It is used for production of eggs that are high in omega-3 and it is now being fed on an experimental basis to Tilapia for the same goal. Camelina microgreens are high in omega-3 fatty acids and can be grown at home for a year around source of fresh greens. They are being introduced into the restaurant industry within kitchen cabinet production systems. Four lines of potatoes that are lower in glycemic index are being analyzed for human consumption experiments aimed at producing a more diabetic friendly potato. A Sentinel Seed Strip is under development for use as an in-garden and on-farm assay of soil nutrient deficiencies. A natural plasmid curing agent(s) is being explored for food security use in reducing the impact of plasmid mediated diseases. OUTCOMES: 1. Advanced testing of low glycemic potatoes for introduction into the retail market; 2. Development of microgreens of Camelina sativa for the home and restaurant market; 3. Development of tests for plasmid curing agents in crop plants; 4. Development of Sentinel Seed Strips to improve the detection of soil minerals deficiencies that lead to foods that are likewise deficient in essential mineral nutrients. OUTCOMES: - Identify low glycemic potato lines for possible value in obesity mitigation; - Establish Camelina microgreens as a source of omega-3 fatty acids. Lower glucosinolate lines are now available; - Establish a Sentinel Seed System to enable gardeners and subsistence farmers to know what elements are deficient in their soil. University of Nebraska at Lincoln (Janos Zempleni): We have focused our research on the following topics: 1) We have discovered 108 proteins in the human proteome that are modified by covalent binding of biotin. Biotinylation of these proteins play roles in cancer prevention, redox biology, and cell stress. 2) We have completed our studies of the roles of holocarboxylase synthetase in orchestrating a multiprotein gene repression complex in human chromatin, and its roles in maintaining genome stability. 3) We have assessed the roles of the FAD (vitamin B2)-dependent histone demethylase LSD1. Our studies suggest that vitamin B2 depletion causes aberrant upregulation of genes in human liver and lymphoid cells. 4) We have developed a high throughput screening assay for assessing the mitochondrial localization of proteins. Using this assay, we are in the process of screening large libraries (collections) of natural and synthetic molecules that prevent the anchoring of acetyl-CoA carboxylase 2 (ACC2) in the mitochondrial membrane. Theoretically, ACC2 inhibitors increase fatty acid oxidation and contribute toward a lean phenotype and metabolic health. 5) We have discovered that humans absorb quantitatively meaningful amounts of microRNAs from nutritionally relevant doses of cow’s milk, milk-borne microRNAs alter gene expression in humans, endogenous microRNA synthesis does not compensate for dietary microRNA depletion, and dietary deficiency may elicit undesirable phenotypes such as bone loss, metabolic syndrome, inflammatory bowel disease, and low reproductive success. Ohio State University (Richard Bruno): We conducted pre-clinical studies to better define anti-inflammatory activities of green tea during NASH and clinical studies examining antioxidant and anti-inflammatory activities of ?-tocopherol on improvements in vascular endothelial function. In a diet-induced obese mouse model, we fed C57BL6 mice either a low-fat or high-fat diet for 8 wk to induce NASH, and then equal numbers of mice from each group were randomized to a low-fat diet or high-fat diet containing green tea extract at 0% or 2% for 8 wk to define the extent to which green tea can effectively serve as a treatment for NASH. Serum markers and histopathology were assessed to determine liver injury. We also examined mRNA expression of NFkB-dependent pro-inflammatory mediators known to contribute to liver injury during NASH. These studies demonstrate that green tea ameliorates obesity otherwise induced by high-fat feeding in association with significant improvements in serum ALT, liver steatosis, and expression of TNF, TNFR1, TLR4, iNOS, but without affecting the expression of Nrf2 or its transcriptional cytoprotective targets. In a clinical study, we conducted a controlled trial in which adults participating in smoking cessation were randomized to co-treatment with placebo or ?-tocopherol prior to evaluating changes in vascular endothelial function using non-invasive ultrasonography and biomarkers of oxidative stress and inflammation. These studies demonstrate that while smoking cessation improves vascular function, co-therapy with ?-tocopherol improves vascular function to a greater extent. The vasoprotective activities occurred without improving several markers of lipid peroxidation, but were significantly correlated with improvements in TNF? and myeloperoxidase, which are known to otherwise impair nitric oxide-dependent vasodilation. In conclusion, these studies support phytochemical-rich diets containing green tea and ?-tocopherol to mitigate pro-inflammatory responses contributing to cardiovascular disease and NASH. Oklahoma State University (Barbara Stoecker): Contribute to the international data on associations between urinary iodine concentrations of pregnant women, their risk for goiter and pregnancy complications, and their knowledge of importance of iodine for human health. Test the effects of zinc supplementation on plasma zinc, a common biomarker of zinc status. Evaluate the assessment of mRNA abundance for selected zinc transporters from PBMC as biomarkers of zinc status and the plausibility of collection of such samples in community settings in Africa. OUTCOMES: Identify the knowledge, attitudes and practices of rural Ethiopian women in relation to the use of iodized salt. Examine the iodine status and cognitive function of primary school children and the knowledge of iodine deficiency of their caretakers. Identify two zinc transporters for which mRNA abundance was decreased by zinc supplementation. Oklahoma State University (Edralin Lucas): The focus of our research group is examining the role of food and their bioactive components in alleviating chronic conditions such as diabetes, obesity, and osteoporosis. For this reporting period, we investigated the effects of 3-month dietary supplementation of freeze-dried mango on glucose parameters, body composition and inflammatory markers in overweight individuals. Our findings indicate that chronic consumption of freeze-dried mango by obese individuals does not negatively impact body weight but modulates fasting blood glucose. We also investigated the effects of freeze-dried watermelon on bone parameters of ovariectomized mice, a model of postmenopausal bone loss. We found that freeze-dried watermelon has modest effect on bone parameters of ovariectomized mice. Oregon State University (Emily Ho): Zinc and chronic disease: Recently we have found that zinc status is compromised with age, possibly due to methylation of the zinc transporter ZIP6. Zinc supplementation reverses age-related zinc deficiency and inhibits age-related inflammatory processes. We have also identified using untargeted metabolomics strategies that methyl-histidine may be a biomarker for human zinc deficiency. Plant-derived phytochemicals and cancer: We have found that sulforaphane, a chemical found in cruciferous vegetables inhibits histone deacetylases, decreases DNA methyl transferase expression, alters histone methylation, lincRNA expression and has anti-cancer properties in the prostate. Bioavailability is decreased in supplemental forms of SFN compared to the whole food. OUTCOMES: 1. Test the effects of zinc status on the gut microbiota, epigenetics, oxidative stress, inflammation, DNA damage and cancer susceptibility in rodent models and humans. 2. Understand the determinants of bioavailability of phytochemicals derived from cruciferous vegetables. 3. Test the ability of sulforaphane supplementation from various sources (supplement vs whole food) to reduce the incidence of prostate cancer via epigenetic modifications. OUTCOMES: -Identify new risk factors in prostate cancer and offer novel dietary modifications to reduce the incidence of prostate cancer; -Establish low cruciferous vegetable intake as a risk factor for the development of prostate cancer by altering histone modifications and cell proliferation pathways; -Gain knowledge of the mechanisms behind the health benefits of micronutrients and phytochemicals such as zinc and compounds derived from cruciferous vegetables;-Establish low dietary zinc as risk factor for inflammatory processes, DNA damage and cancer risk and identify new biomarkers for human zinc deficiency; -Establish function of zinc and changes in zinc metabolism with age. Pennsylvania State University (Jairam Vanamala): Anti-inflammatory activity of purple-fleshed potatoes: Recently we have found that only purple-fleshed potatoes, even after processing, prevent and reverse high-fat diet induced colonic-systemic inflammation and oxidative stress markers in pig model. Anti-cancer activity of purple-fleshed potatoes: We have found that purple-fleshed potato extracts that are high in anthocyanins decrease in vitro human colon cancer stem cell proliferation, elevate apoptosis and suppresses cytoplasmic and nuclear levels of ?-catenin, the critical downstream effector of Wnt pathway. We have also shown that purple-fleshed potatoes, even after baking, significantly suppress colon tumor number, colonocyte proliferation (ki-67; immunofluorescence) and concomitantly suppressed nuclear beta-catenin expressing oncogenic stem cells (lgr5 positive) in rodent-model of colon carcinogenesis. OUTCOMES: 1. Test the effect of purple-fleshed potatoes against high-fat diet induced colonic-systemic inflammation using human-relevant, agriculturally important pig model. 2. Test the anti-cancer activity of purple-fleshed potatoes against human colon cancer stem cells in vitro and AOM-induced colon tumorigenesis in rodent models. OUTCOMES: -Determine the effect of processing on anti-inflammatory and antioxidant activity of anthocyanin containing color-fleshed potatoes;-Determine whether processed color-fleshed potatoes prevent and reverse the high-fat diet induced colonic inflammatory markers via alteration in gut bacteria and metabolism; -Determine whether color-fleshed potatoes suppress colon cancer via suppression of colon cancer stem cells. Purdue University (Connie Weaver): This year our laboratory studied the effect of several bioactive ingredients or foods on bone health including hesperidin and grape powder using calcium tracer urinary appearance from pre-labeled bone and more traditional measures. Grape produced a modest advantage to bone in growing rats. Calcium, but not hesperidin, increased net bone 41Ca retention in postmenopausal women.

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