W3002: Nutrient Bioavailability--Phytonutrients and Beyond

(Multistate Research Project)

Status: Inactive/Terminating

Project Menu

SAES-422 Reports

Annual/Termination Reports:

[07/29/2014] [10/05/2015] [07/19/2016] [06/19/2017] [04/02/2018]

Date of Annual Report: 07/29/2014

Report Information

Annual Meeting Dates: 05/29/2014 - 05/30/2014
Period the Report Covers: 10/01/2013 - 09/01/2014

Participants

Weaver, Connie (Meeting Chair) (weavercm@purdue.edu) – Purdue University;
Zempleni, Janos (jzempleni2@unl.edu) – University of Nebraska, Lincoln;
Stoecker, Barbara (Barbara.stoecker@okstate.edu) – Oklahoma State University;
Bruno, Rich (Bruno.27@osu.edu) – Ohio State University;
Ho, Emily (emily.ho@oregonstate.edu) – Oregon State University;
Bray, Tammy (tammy.bray@oregonstate.edu) – Oregon State University;
Andrade, Juan (jandrade@illinois.edu) – University of Illinois;
Lindshield, Brian (blindsh@ksu.edu) – Kansas State University;
Park, Yeonhwa (ypark@foodsci.umass.edu) – University of Massachusetts;
Sands, David (uplds@montana.edu) – Montana State University;
Teske, Jennifer (teskeja@email.arizona.edu) – University of Arizona;
Vanamala, Jairam (juv4@psu.edu) – Pennsylvania State University;
Chester, Deirdra (dnchester@nifa.USDA.gov) – USDA Representative (via Skype);

Brief Summary of Minutes

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

Accomplishments

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.

Publications

Abebe, Howit, Abebe, Yewelsew, Loha, Eskinder, Stoecker BJ. (2014) Consumption of vitamin A rich foods and dark adaptation threshold of pregnant women at Damot Sore District, Wolayita, Southern Ethiopia. Ethiop J Health Sci (In Press). Amer, F.S., L. Reddivari, G. P. Madiwale, M. Stone, D. G. Holm and J. Vanamala. 2014. Effect of genotype and storage on glycoalkaloid and acrylamide content and sensory attributes of potato chips. American Journal of Potato Research. In Press. Andrade, J.E., Burgess, J.R. 2013. Effect of Dietary EGCG on Normal and Vitamin E and Selenium Deficient Rats. Food & Nutr Sci. 4(8): 163-173 [DOI: 10.4236/fns.2013.48A020] Andrade, J.E., Rosales, E., Lopez, J.R., Carrillo, E.P., Engeseth, N.J., and W.G. Helferich. 2014. Development of a Point-of-Use Fortification Technology for Delivery of Micronutrients in Honduras. J Sci Food Agric. [In Press] [DOI: 10.1002/jsfa.6736] Andrade, J.E., Young, J., Baker, C., Doerge, D., W.G. Helferich. Long-term dietary exposure to dietary of genistein alone or within soy protein isolate affects the progression of human breast cancer (MCF-7) cells after genistein withdrawal. Molec Nut Food Res. 2014 [DOI: 10.1002/mnfr.201300780] Baier SR, Nguyen C, Xie F, Wood JR, Zempleni J. MicroRNA from cow’s milk are bioavailable and affect gene expression in humans (submitted) Baier SR, Zbasnik R, Schlegel V, Zempleni J. Off target effects of sulforaphane include the de-repression of long-terminal repeats through histone acetylation . J Nutr Biochem 25: 665-668, 2014 KD Ballard, BR Kupchak, BM Volk, A Shkreta, C Liptak, AS Ptolemy, E Mah, MS Kellogg, RS Bruno, RL Seip, CM Maresh, WJ Kraemer, JS Volek. (2013). Acute effects of ingestion of a novel whey-derived extract on vascular endothelial function in overweight, middle-aged men and women. Br J Nutr, 13:109(5):882-93. KD Ballard, E Mah, Y Guo, R Pei, JS Volek, RS Bruno. (2013). Low-fat milk ingestion prevents postprandial hyperglycemia-mediated impairments in vascular endothelial function in obese individuals with metabolic syndrome. J Nutr, 143(10):1602-10. Baltrus D., Yourstone S., Lind A., Guilbaud C., Sands D.C., Jones C.D., Morris C.E., Dangl J.L. 2014. Draft genome sequences of a phylogenetically diverse suite of Pseudomonas syringae strains from multiple source populations. Genome Announcements 2(1):e01195-13. doi:10.1128/genomeA.01195-13. Bartoli C., Berge O., Monteil C.L., Guilbaud C., Balestra G.M., Varvaro L., Jones C., Dangl J., Baltrus D., Sands D.C., Morris C.E. 2014. The Pseudomonas viridiflava phylogroups of the P. syringae species complex are characterized by genetic variability and phenotypic plasticity of pathogenicity-related traits. Environ Microbiol doi: 10-1111/1462-2920.12433 Beaver, LM, Yu, T., Sokolowoski, EI, Williams, DE, Dashwood, RH and Ho, E. (2013) Chemopreventative phytochemical 3,3’-diindolylmethane inhibits histone deacetylases in prostate cancer cells. Tox Appl Pharm 263:345-51. Bekele, Genet. (2013) Dietary Vitamin A Consumption and Associated Factors in 12-59 Month Old Children at Tula, SNNPR. M.Sc. Thesis. Hawassa University. T. Benn, B. Kim, Y. Park, C. S. Ku, Y. Yang, T. X. Pham, C. Wegner, C. Farruggia, E. Harness, J. Lee. Supplementation of polyphenol-rich blackcurrant extract exerted hypolipidemic and anti-inflammatory effect in diet-induced obese mice. FASEB J 2014; (Abstract). T. Benn, B. Kim, Y. Park, C. J. Wegner, E. Harness, T-G. Nam, D-O. Kim, J. S. Lee, J. Lee. Polyphenol-rich blackcurrant extract prevents inflammation in diet-induced obesity mice. J Nutr Biochem 2014; (In Press). Bhattacharyya M, Weaver C. Calcium isolation from large-volume human urine samples for 41Ca analysis by accelerator mass spectrometry. Appl Rad Isotopes 2013 78:57-61. Chen, JJ., Zheng, JK., McClements, D.J., Xiao, H. Tangeretin-loaded protein nanoparticles fabricated from zein/beta-lactoglobulin: Preparation, characterization, and functional performance. Food Chem., 2014 (In press) H. T. Cho, L. Salvia-Trujillo, J. Kim, Y. Park, H. Xiao, and D. J. McClements (2014) Influence of Droplet Size and Composition on Bioavailability of Long Chain Fatty Acids and Coenzyme Q10: An In Vivo Study, Food Chem. 156:117-122. Ersino G, Tadele H, *Bogale A, Abuye C, Stoecker BJ. (2013) Clinical assessment of goiter and low urinary iodine concentration depict presence of severe iodine deficiency in pregnant Ethiopian women: a cross-sectional study in rural Sidama, southern Ethiopia. Ethiop Med J 51:133-141. Evans SF, Meister M, Peterson S, Perkins-Veazie P, Clarke SL, Payton M, Smith BJ, Lucas EA. Mango supplementation improves blood glucose in obese individuals. Submitted to Nutrition and Metabolic Insights (accepted pending revisions). Feresin R, Johnson S, Elam M, Kim J, Khalil D, Lucas EA, Smith B, Payton ME, Akhter M, Arjmandi BH. Effects of vitamin E on bone biomechanical and histomorphometric parameters in ovariectomized rats. J Osteopor. 2013 Gebreegziabher T, Stoecker BJ. (2013) Vitamin D insufficiency in a sunshine sufficient area: Southern Ethiopia. Food Nutr Bull 34:429-433. Gebreegziabher Tafere, Teyikie N, Mulugeta A, Abebe Y, Hambidge KM, Stoecker BJ. (2013) Lack of dietary sources of iodine and the prevalence of iodine deficiency in rural women from Sidama zone, southern Ethiopia. African J Food, Nutr Develop 13:8401-8414. Grider, A, Wickwire, K, Ho, E, Chung, CS and King, J. (2013) Dietary zinc depletion and repletion affects plasma proteins: an analysis of the plasma proteome. Biometals 26(1):133-40. Y Guo, E Mah, CG Davis, T Jalili, MG Ferruzzi, OK Chun, RS Bruno. (2013). Dietary fat increases quercetin bioavailability in overweight adults. Mol Nutr Food Res, 57(5):896-905. Henley, T, Reddivari, L, Broeckling, C, Bunning, M, Miller, J, Avens, J, Stone, M, Prenni, J, Vanamala, J. 2014. American India Pale Ale matrix rich in xanthohumol is potent in suppressing proliferation and elevating apoptosis of human colon cancer cells. International Journal of Food Science and Technology, In Press. Iwaniec UT, Turner RT, Smith BJ, Stoecker BJ, Rust A, Zhang B, Vasu VT, Gohil K, Cross CE, Traber MG. (2013) Evaluation of long-term vitamin E insufficiency or excess on bone mass, density and microarchitecture in rodents. Free Radic Biol Med 65:1209-14. Kandianis CB, Stevens R, Liu W, Palacios N, Montgomery K, Pixley K, White WS, Rocheford T. Genetic architecture controlling variation in grain carotenoid composition and concentrations in two maize populations. Theor Appl Genet 2013;126:2879-95. Kaur P, Shorey LE, Ho E, Dashwood RH, Williams DE. (2013) The epigenome as a potential mediator of cancer and disease prevention in prenatal development. Nutr Rev. 71(7):441-57. B. Kim, S. Lee, Y. Park, C. S. Ku, T. X. Pham, C. J. Wegner, Y. Yang, S. I. Koo, O. K. Chun, J. Lee. Blueberry, blackberry, and blackcurrant differentially affect plasma lipids and pro-inflammatory markers in diet-induced obesity mice. Eur J Nutr 2014; (In review). B. Kim, Y. Park, S. I. Koo, J. Lee. Role of polyphenol-rich blackcurrant and black chokeberry extracts in the stimulation of transintestinal cholesterol excretion in vitro. FASEB J 2014; (Abstract). LJ Kunces, LK Cusack, BR Kupchak, BM Volk, DJ Freidenreich, JC Aristizabal, C Saenz, R Pei, Y Guo, ML Fernandez, RS Bruno, CM Maresh, WJ Kraemer, A Pronczuk, KC Hayes, JS Volek. (2013). Triglyceride recrystallized phytosterols in fat-free milk improve lipoprotein profiles more than unmodified free phytosterols in hypercholesterolemic men and women. J Am Coll Nutr, 32(4):234-42. S. Lee, B. Kim, Y. Yang, T. X. Pham, Y. Park, J. Manatou, S. I. Koo, O. K. Chun, J. Lee. Berry anthocyanins suppress the expression and secretin of pro-inflammatory mediators in macrophages by inhibiting nuclear translocation of NF-B independent of NRF2-mediated mechanism. J Nutr Biochem 2014; 25:404-411. (PMID: 24565673) Legette LL, Prasain J, King J, Arabshahi A, Barnes S, Weaver CM. Pharmacokinetics of equol, a soy isoflavone metabolite, changes with the form of equol (dietary versus intestinal production) in ovariectomized rats. J Ag Food Chem 62:1264-1300, 2014. Lelievre SA, Weaver CM. Global nutrition research: nutrition and breast cancer prevention as a model. Nutr Rev doi:10.1111/nure.12075 70:1-11, 2013. Lewis RD, Liang EM, Hill Gallant KM, Hall DB, McCabe GP, Hausman DB, Martin BR, Warden SJ, Peacock M, Weaver CM. A randomized trial of vitamin D3 supplementation in children: Dose-response effects on vitamin D metabolites and calcium absorption. J Clin Endocrin Med 98:4816-4825, 2013. Li Y, Hassan YI, Moriyama H, Zempleni J. Holocarboxylase synthetase interacts physically with euchromatic histone-lysine N-methyltransferase, linking histone biotinylation with methylation events. J Nutr Biochem 24:1446-1452, 2013 Li Y, Malkaram SA, Zhou J, Zempleni J. Lysine biotinylation and methionine oxidation in the heat shock protein HSP60 synergize in the elimination of reactive oxygen species in human cell cultures. J Nutr Biochem (in press) Lipkie TE, Janaschb A, Cooperb JR, Hohman EE, Weaver CM, Ferruzzi MG. Quantification of vitamin D and 25-hydroxyvitamin D in soft tissues by liquid chromatography-tandem mass spectrometry. J Chromatography B 932:6-11, 2013. Liu D, Zempleni J. Holocarboxylase synthetase interacts physically with the nuclear receptor corepressor, histone deacetylase 1, and a novel splicing variant of histone deacteylase 1 to repress repeats. (submitted) Liu D, Zempleni J. Low activity of LSD1 elicits a pro-inflammatory gene expression profile in riboflavin-deficient human T lymphoma Jurkat cells. (submitted) Liu D, Zempleni J. Transcriptional regulation of the albumin gene depends on the removal of histone methylation marks by the FAD-dependent monoamine oxidase LSD1 in HepG2 human hepatocarcinoma cells. J Nutr (in press) Luizzi, JP, Wong, CP, Ho, E and Tracy, A. (2013) Regulation of hepatic suppressor of cytokine signaling 3 (SOCS3) by zinc. J. Nutr Biochem, Jun;24(6):1028-33 E Mah, SK Noh, KD Ballard, HJ Park, JS Volek, RS Bruno. (2013). Supplementation of a ?-tocopherol-rich mixture of tocopherols in healthy men protects against vascular endothelial dysfunction induced by postprandial hyperglycemia. J Nutr Biochem. 24(1):196-203. E Mah, R Pei, Y Guo, KD Ballard, T Barker, VE Rogers, BA Parker, AW Taylor, MG Traber, JS Volek, RS Bruno. (2013) ?-Tocopherol-rich supplementation additively improves vascular endothelial function during smoking cessation. Free Radic Biol Med, 65: 1291-1299. KM Mangano, HL Hutchins-Wiese, AM Kenny, SJ Walsh, RH Abourizk, RS Bruno, R Lipcius, P Fall, A Kleppinger, L Kenyon-Pesce, KM Prestwood, JE Kerstetter. (2013). Soy proteins and isoflavones reduce interleukin-6 but not serum lipids in older women: a randomized controlled trial. Nutr Res, 33(12):1026-33. Mao, Y.Y., M. Dubot, H. Xiao, and D.J. McClements, Interfacial Engineering Using Mixed Protein Systems: Emulsion-Based Delivery Systems for Encapsulation and Stabilization of beta-Carotene. Journal of Agricultural and Food Chemistry, 2013. 61(21): p. 5163-5169. Maria, K, Radhakrishnan, S, Reddivari, L, Vanamala, J. 2014. Colon Carcinogenesis: Influence of Western Diet-induced Obesity and Targeting Stem Cells Using Dietary Bioactive Compounds. Nutrition, In Press. Massey AR, Reddivari L, Vanamala J. 2014. The Dermal Layer of Sweet Sorghum (Sorghum bicolor) Stalk, a Byproduct of Biofuel Production and Source of Unique 3-Deoxyanthocyanidins, has more Anti-proliferative and Pro-apoptotic Activity than Pith in p53 Variants of HCT116 and Colon Cancer Stem Cells. Journal of Agriculture and Food Chemistry, 62:3150-3159. C Masterjohn, E Mah, Y Park, R Pei, J Lee, JE Manautou, RS Bruno. (2013). Acute glutathione depletion induces hepatic methylglyoxal accumulation by impairing its detoxification to D-lactate. Exp Biol Med, 238(4):360-9. C Masterjohn, Y-K Park, J-Y Lee, SK Noh, SI Koo, RS Bruno. (2013). Dietary fructose feeding increases adipose methylglyoxal accumulation in association with low expression and activity of glyoxalase-2. Nutrients, 5, 3311-28. Mavanji V*, Teske JA*, Billington CJ and Kotz CM. Partial sleep deprivation by environmental noise increases food intake and body weight in obesity resistant rats. Obesity (Silver Spring). 21(7):1296-405, 2013. Epub 2013 May 13. McClements, D.J., Xiao, H. Excipient foods: designing food matrices that improve the oral bioavailability of pharmaceuticals and nutraceuticals. Food & Funct. 2014, DOI: 10.1039/c4fo00100a. McNay EC, Teske JA, Kotz CM, Dunn-Meynell A, Levin B, McCrimmon RJ and Sherwin RS. Long-term, intermittent, insulin-induced hypoglycemia produces marked obesity in the absence of hyperphagia or insulin resistance: a model for weight gain associated with intensive insulin therapy. American Journal of Physiology Endocrinology and Metabolism. 304(2):E131-8, 2013. Meyer M, Kesic MJ, Clarke J, Ho E, Simmen RC, Diaz-Sanchez D, Noah TL, Jaspers I. (2013) Sulforaphane induces SLPI secretion in the nasal mucosa. Respir Med 107(3):472-5. Molloy, AM, Einri, CN, Jain, D, Laird, E, Fan, R, Wang, Y, Scott, JM, Shane, B, Brody, LC, Kirke, PN, and Mills, JL. Is Low Iron Status a Risk Factor for Neural Tube Defects? Birth Defects Research (Part A) Clin Mol Teratol 100:100–106, 2014. Morris C.E., Conen F., Huffman J.A., Phillips V., Pöschl U., Sands D.C. 2014. Bioprecipitation: A feedback cycle linking Earth history, ecosystem dynamics and land use through biological ice nucleators in the atmosphere. Global Change Biology 20:341-351 (doi: 10.1111/gcb.12447) Morris C.E., Sands D.C., Glaux C., Samsatly J., Asaad S., Moukahel A.R., Gonçalves F.L.T., Bigg E.K. 2013. Urediospores of rust fungi are ice nucleation active at > ?10 °C and harbor ice nucleation active bacteria. Atmos. Phys. Chem. 13:4223-4233. Palacios C, Martin BR, McCabe GP, McCabe L, Peacock M, Weaver CM. Dietary calcium requirements do not differ between Mexican American boys and girls. J Nutr In press, 2014. Park CY, Lee WH, Fleet JC, Allen MR, McCabe GP, Walsh DM, Weaver CM. Calcium and vitamin D intake maintained from pre-ovariectomy independently affect calcium metabolism and bone properties in Sprague Dawley rats. J Nutr In press. Pawlowski J, Martin B, McCabe G, Ferruzzi M, Weaver, C. Polyphenolic-rich extracts of plum, blueberry, grape seed, soy, and grape improve bone calcium retention in ovariectomized Sprague Dawley rats. J Ag Food Chem. In press. Peterson, R.D., Cunnigham, B.T., Andrade, J.E. A photonic crystal biosensor assay for ferritin utilizing iron-oxide nanoparticles. Biosens Bioelect. 2014 [DOI: 10.1016/j.bios.2014.01.022] Rajendran P, Kidane AI, Yu TW, Dashwood WM, Bisson WH, Löhr CV, Ho E, Williams DE, Dashwood RH. (2013) HDAC turnover, CtIP acetylation and dysregulated DNA damage signaling in colon cancer cells treated with sulforaphane and related dietary isothiocyanates. Epigenetics. 8(6):612-23. Romagnolo DF, Zempleni J, Selmin OI. Nuclear receptors and epigenetic regulation: opportunities for nutritional targeting and disease prevention. Adv Nutr 5:373-385, 2014 DC Sands, A Pilgeram, CE Morris. 2014. Marketing of perennial grains with benefits for human health and nutrition. In: Development of Perennial Crops for food Security: Proceedings of the FAO Expert Workshop. (http://www.fao.org/3/a-i3495e.pdf). Shaltiel G, Bar-David E, Meiron OE, Waltman E, Shechter A, Aflalo ED, Stepensky D, Berman A, Martin BR, Weaver CM, Sagi A. Bone loss prevention in ovariectomized rats using stable amorphous calcium carbonate. Health, Special Issue: New and Emerging Therapies for Osteoporosis. Health 5(7A2):18-29, 2013. Shane, B. (2013). Folate and vitamin B12 function. In Encyclopedia of Biological Chemistry, Lennarz, W. and Lane, M. D., eds., 2nd ed., pp. 324-328, Elsevier, New York. Shorey LE, Madeen EP, Atwell LL, Ho E, Löhr CV, Pereira CB, Dashwood RH, Williams DE. (2013). Differential modulation of dibenzo[def,p]chrysene transplacental carcinogenesis: maternal diets rich in indole-3-carbinol versus sulforaphane. Toxicol Appl Pharmacol. 270(1):60-9. Smith BJ, Bu SY, Wang Y, Rendina E, Lim YF, Marlow D, Clarke SL, Cullen DM, Lucas EA. A comparative study of the bone metabolic response to dried plum supplementation and PTH treatment in adult, osteopenic ovariectomized rat. Bone 2014;58:151-9. Smith BJ, Graef JL, Rendina E, Clark KA, Clarke SL, Lucas EA, Wronski TJ, Halloran BP. Effects of dried plum supplementation on bone metabolism in adult C57BL/6 male mice. Calcif Tissue Int. 2014 Apr;94(4):442-53. Teske JA, Billington CJ and Kotz CM. Mechanisms underlying obesity resistance associated with high spontaneous physical activity. Neuroscience. 256: 91-100. 2013. Epub 2013 Oct. 22. Teske JA, Perez-Leighton C, Billington CJ and Kotz CM. Role of the locus coerulues in enhanced orexin A-induced spontaneous physical activity in obesity resistant rats. American Journal of Physiology Regulatory, Integrative and Comparative Physiology. 305: R1337-R1345. 2013. Epub 2013 Oct. 2. Ursell LK, Haiser HJ, Van Treuren W, Garg N, Reddivari L, Vanamala J, Dorrestein PC, Turnbaugh PJ, Knight R. 2014. The Intestinal Metabolome: An Intersection Between Microbiota and Host. Gastroenterology, 146(6): 1470-1476. Vidya S. Farook, Lavanya Reddivari, Geetha Chittoor, Sobha Puppala, Rector Arya, Sharon P. Fowler, Kelly J. Hunt, Joanne E. Curran, Anthony G. Comuzzie, Donna M. Lehman, Christopher P. Jenkinson, Jane L. Lynch, Ralph A. DeFronzo, John Blangero, Daniel E. Hale, Ravindranath Duggirala, Jairam Vanamala. 2014. Metabolites as Novel Biomarkers for Childhood Obesity-Related Traits in Mexican American Children. Pediatric Nutrition. Accepted. JS Volek, BM Volk, AL Gómez, LJ Kunces, BR Kupchak, DJ Freidenreich, JC Aristizabal, C Saenz, C Dunn-Lewis, KD Ballard, EE Quann, DL Kawiecki, SD Flanagan, BA Comstock, MS Fragala, JE Earp, ML Fernandez, RS Bruno, AS Ptolemy, MD Kellogg, CM Maresh, WJ Kraemer. (2013). Whey protein supplementation during resistance training augments lean body mass. J Amer Coll Nutr, 32(2):122-35. Wang, R, Lohr, C., Fischer, KA, Dashwood, WM, Greenwood, J, Ashktorab, H, Ho, E., Williams, DE, Dashwood, MR and Dashwood, RH. (2013). Epigenetic inactivation of endothelin-2 (ET-2) and ET-3 in colon cancer. Int J Cancer, 132(5):1004-12 Warden SJ, Hill KM, Ferira AJ, Laing EM, Martin BR, Hausman DB, Weaver CM, Peacock M, Lewis RD. Racial differences in cortical bone and biochemical variables in black and white children in the early stages of puberty. Osteoporos Int 24:1869-79, 2013 Watson GW, Beaver LM, Williams DE, Dashwood RH, Ho E. (2013) Phytochemicals from Cruciferous Vegetables, Epigenetics, and Prostate Cancer Prevention. AAPS J. 15(4):951-61 Weaver CM. A personal perspective on discoveries at the interface of food science and nutrition. Nutr Today 48:241-244, 2013. Weaver CM. Bioactive foods and ingredients for health. Adv Nutr 5:306S-311S, 2014. Weaver CM. Calcium Supplementation: Is protecting against osteoporosis counter to protecting against cardiovascular disease. Current Osteoporosis Reports DOI 10.1007/s11914-014-0208-1, 2014. Weaver CM. How sound is the science behind the dietary recommendations for dairy. Am J Clin Nutr doi: 10.3945/ajcn.113.073007, 2014. Weaver CM, Dwyer J, Fulgoni V, King JC, Leveille GA, MacDonald RS, Ordovas J, Schnakenberg D. ASN Scientific Statement: Processed foods: Contributions to Nutrition. AJCN, In press, 2014. Weaver, C.M., Heaney, R.P. Ch. 42 Nutrition and Osteoporosis In: Primer on the Metabolic Bone Diseases and Disorders of Mineral Metabolism, 8th Ed. Rosen, C.J. ed. American Society for Bone and Mineral Research. Wiley-Blackwell Washington DC Pp 361-366 2013. Weaver CM, Wijeshinha-Bettoni R, McMahon D, Spence LA. Ch 4. Milk and dairy products as part of the diet. In: Milk and Dairy Products in Human Nutrition. Food and Agriculture Organization of the United Nations (FAO), Rome Italy. Muehlhoff E, Bennett A, McMahon D, Eds, pg 103-182, 2013. Whisner CM, Martin BR, Nakatsu CH, McCabe GP, McCabe LD, Peacock M, Weaver CM. Soluble corn fiber affects short-term calcium absorption in adolescent boys and girls: A randomized controlled trial using dual stable isotopic tracers, Br J Nutr In press. Whisner CM, Martin BR, Schoterman MHC, Nakatsu CH, McCabe LD, McCabe GP, Wastney ME, van den Heuvel EGHM, Weaver CM. Galacto-oligosaccharides increase calcium absorption and gut bifidobacteria in young girls: A double blind crossover trial. Br J Nutr 110:1292-1303, 2013. Wong, CP, Hsu, A, Buchanan, A, Palomera-Sanchez, Z, Beaver, LM, Houseman, EA, Williams, DE, Dashwood, RH and Ho, E. (2014) Effects of sulforaphane and 3,3'-diindolylmethane on genome-wide promoter methylation in normal prostate epithelial cells and prostate cancer cells. PLOS One, 9(1):e86787. doi: 10.1371/journal.pone.0086787 Wong, CP, Magnusson, KR and Ho, E (2013). Increased inflammatory response with age is associated with age-related zinc deficiency and zinc transporter dysregulation. J.Nutr Biochem, 24:353-9. Xia M, Malkaram SA, Zempleni J. Three promoters regulate the transcriptional activity of the human holocarboxylase synthetase gene. J Nutr Biochem 24: 1963-1969, 2013 Xue J, Wijeratne SS, Zempleni J. Holocarboxylase synthetase synergizes with methyl CpG binding protein 2 and DNA methyl transferase 1 in the transcriptional repression of long terminal repeats. Epigenetics 8:504-511, 2013 Xue J, Zempleni J. Epigenetic synergies among biotin, folate, and holocarboxylase synthetase in the regulation of pro-inflammatory cytokines and repeats. Scand J Immunol 78: 419-425, 2013 Xue J, Zhou J, Zempleni J. Holocaboxylase synthetase catalyzes biotinylation of heat shock protein 72, thereby inducing RANTES expression in HEK293 cells. Am J Physiol Cell Physiol 305: C1240-C1245, 2013 Yao, M., Chen, J., Zheng, J., Song, M., McClements, D.J., Xiao, H. Enhanced lymphatic transport of bioactive lipids: Cell culture study of polymethoxyflavone incorporation into chylomicrons. Food & Funct., 2013, DOI: 10.1039/C3FO60335K Yao, M., Xiao, H., McClements, D.J. Delivery of lipophilic bioactives: assembly, disassembly, and reassembly of lipid nanoparticles, Annu. Rev. Food Sci. Technol. 2014;5:53-81 doi: 10.1146/annurev-food-072913-100350. Zempleni J, Barshop B, Cordonier EL, Baier SR, Gertsman, I. Disorders of biotin metabolism. In: The Online Metabolic & Molecular Bases of Inherited Disease. Editors: Valle, Baudet, Vogelstein, Kinzler, Antonarakis, Ballabio, Scriver (Emeritus), Childs (Emeritus), Sly (Emeritus), Bunz (Parts Editor), Gibson (Parts Editor), Mitchell (Parts Editor). McGraw Hill. Zempleni J, Cordonier EL, Baier SR, Xue J. Vitamins, Bioactive Food Compounds, and Histone Modifications. In: Handbook of Vitamins. Zempleni J, Suttie JW, Gregory JF III, Stover PJ (eds), 5th edition. Taylor and Francis, Inc., Boca Raton, FL, 2013. Zempleni, J, Eng WK, Singh MP, Baier SR. The chemistry and biochemistry of biotin. In: Food and Nutritional Components in Focus. Preedy VR (ed.), Royal Society of Chemistry, London, U.K., 2013, pp. 146-157 Zempleni J, Liu D. Camara DT, Cordonier EL. Novel roles of holocarboxylase synthetase in gene regulation and intermediary metabolism. Nutr Rev 72(6):369-76, 2014 Zempleni J, Liu D, Camara Teixeira D, Singh MP. Mechanisms of gene transcriptional regulation through biotin and biotin-binding proteins in mammals. In: Vitamin-binding Proteins – Their Functional Consequences. Dakshinamurti D, Dakshinamurti S (eds.), Taylor & Francis, Boca Raton, FL, 2013 pp. 219-228 Zempleni J, Liu D, Xue J. Nutrition, histone epigenetic marks, and disease. In: Epigenomics in Health and Disease. Jirtle, RL, Tyson F (eds.), Springer, Heidelberg, Germany, 2013, pp. 197-217 Zheng, JK., Li, Y., Song, MY., Fang, X., Cao, Y., McClements, D.J., Xiao, H. Improving Intracellular Uptake of 5-Demethyltangeretin by Food Grade Nanoemulsions. Food Research International. 2014 (in press).

Impact Statements

Insufficient sleep promotes excessive energy intake and fatigue, which reduces energy expenditure and increases risk for bacterial infection, obesity, insomnia, personal injury and reduced quality of life. University of Arizona, Jennifer A. Teske
Recently, there has been concern about increased cancer risk and exacerbation of B12 deficiency by folate fortification. The models we have developed may indicate whether disease risk can be modified by dietary changes such as folate fortification. Our recent studies do not support any adverse effects of folate fortification on vitamin B12 status in a young population. Our genetic studies may suggest novel biomarkers for assessing vitamin status. University of California, Berkeley, Barry Shane
Polyphenols found in berries regulate lipid metabolism and exert anti-inflammatory effects, which are beneficial to prevent obesity-associated chronic inflammation and other metabolic diseases. University of Connecticut, Ji-Young Lee
Biosensor. A proof-of-concept platform using photonic crystal technology for detection and quantification of ferritin and soluble transferrin receptor was developed. Biosensor allows for accurate and sensitive detection of iron biomarkers in serum similar to commercial ELISAs. University of Illinois, Urbana-Champaign, Juan Andrade
Fortification. A pilot scale process, similar to Guatemalan?s, for the addition of chelated iron mixed in an extruded corn:rice puff to nixtamalized corn was achieved after wet grinding. It showed uniform distribution of chelated iron at different levels after simple mixing and kneading. University of Illinois, Urbana-Champaign, Juan Andrade
Education. The International Service and Experiential Learning Program in Honduras had six undergraduate students. The program had the following activities: a) cultural exchange of students with rural families in St. Lucia valley; b) service providing anthropometric measurements in rural schools; c) research related to food security, dietary diversity and physical activity; d) experiential learning of food systems at Zamorano University. University of Illinois, Urbana-Champaign, Juan Andrade
Our research helped define the role of the amount of added vegetable oil as a limiting factor in bioavailability of carotenoids/fat-soluble vitamins from fresh vegetables. Our findings resulted in a bioavailability claim on the Wish-bone® salad dressing label: ?Oils like those in Wish-bone® help better absorb vitamins A & E from salad.? This helped increase profits for the US company and educated consumers about benefits of vegetable oil in a healthy diet. Iowa State University, Wendy White
We expect to generate data on the new fortified blended foods that will help determine whether they are suitable alternatives to corn-soy blend for use in food aid. We plan to present our results at conferences and publish them in peer-reviewed journals. If our results meet our expectations, the FBFs we developed being used for food aid. Kansas State University, Brian Lindshield
Functional foods with improved health benefits can be improved by developing food-grade structured delivery systems. University of Massachusetts, Yeonhwa Park
Plant breeding is generally focused on yield and agronomic characteristics. This research is significant because the inclusion of nutritional traits are in plant breeding and can lead to more nutritious foods. Mineral deficiency in the soil translates into mineral deficient foods produced in such soils. Increasing awareness of deficiencies using selected barley indicator mutant lines to detect mineral deficient soils, is one advance in this area. Montana State University, David Sands
Our research includes collaborations with 14 scientists. Thirteen graduate students, undergraduate students, and staff were advised and supervised. Nineteen papers were published and 29 presentations were delivered. A website is hosted that disseminates information related to W3002 activities (nutrigenomics.unl.edu). University of Nebraska at Lincoln, Janos Zempleni
Clinical studies indicate ?-tocopherol supplementation during short-term smoking cessation improves vascular endothelial function to a greater extent than smoking cessation alone by lowering inflammation. Pre-clinical studies in an obese rodent model demonstrate green tea protects against hepatic inflammation otherwise contributing to liver injury during NASH. Use of these dietary constituents may lower incidence of chronic disease mediated by inflammation. Ohio State University, Richard Bruno
Working with iodine deficient populations in Ethiopia allowed us to provide behavior change communication but also to develop the local evidence-base to encourage policy makers to implement salt iodization programs for the country. Additionally, work in a population group with chronically low dietary zinc intakes has provided the opportunity to assess additional biomarkers that might be useful in assessment of zinc status in various population groups. Oklahoma State University, Barbara Stoecker
Obesity is a major health problem worldwide and is associated with many chronic conditions such as diabetes, heart disease and osteoporosis. Mango supplementation may offer an innovative dietary intervention in modulating blood glucose without negative effects on body composition. Watermelon provides many bioactive compounds such as lycopene, ?-carotene, vitamin C, cucurbitacin E, and citrulline, and shows modest health-promoting properties. Oklahoma State University, Edralin Lucas
Diet plays an important role in mitigating the development and progression of several cancers, including prostate. This research demonstrates that nutritional strategies that decrease oxidative stress, inflammation, DNA damage and/or target aberrant epigenetic alterations, such as acetylation and methylation, in prostate cancer have the potential to dramatically reduce the incidence of prostate cancer Oregon State University, Emily Ho
Diet plays a critical role in colonic oxidative stress and inflammation. This research is significant because color-fleshed potatoes suppress oxidative stress and inflammatory markers in pigs as well as colon cancer stem cells in mice. This work will form the basis for future clinical trials to identify the evidence-based whole food prevention/therapeutic strategies against inflammatory bowel disease and colon cancer. Pennsylvania State University, Jairam Vanamala
Dietary approaches to reducing bone loss promise fewer side effects compared to osteoporosis therapies and are more in line with long term prevention approaches. Purdue University, Connie Weaver

Date of Annual Report: 10/05/2015

Report Information

Annual Meeting Dates: 08/05/2015 - 08/06/2015
Period the Report Covers: 10/01/2014 - 09/01/2015

Participants

Sands, David (Meeting Chair) (davidsands41@yahoo.com) – Montana State University; Andrade, Juan (Meeting Secretary) (jandrade@illinois.edu) – University of Illinois, Urbana-Champaign; Bray, Tammy (Admin. Advisor) (tammy.bray@oregonstate.edu) – Oregon State University; Bruno, Richard (Bruno.27@osu.edu) – Ohio State University; Ho, Emily (emily.ho@oregonstate.edu) – Oregon State University; Lee, Ji-Young (ji-young.lee@uconn.edu) – University of Connecticut; Lin, Dingbo (Daniel) dingbo.lin@okstate.edu)- Oklahoma State University (new-W3002 participant); Lindshield, Brian (blindsh@ksu.edu) – Kansas State University; Park, Yeonhwa (ypark@foodsci.umass.edu) – University of Massachusetts; Stoecker, Barbara (barbara.stoecker@okstate.edu) – Oklahoma State University; Teske, Jennifer (teskeja@email.arizona.edu) – University of Arizona; Weaver, Connie (weacercm@purdue.edu) – Purdue University, West Lafayette; Chester, Deirdra (dnchester@nifa.USDA.gov) - USDA Representative (via Skype)

Brief Summary of Minutes

Meeting was called to order August 5, 2015 at 8:00 a.m. MST.

Welcome and introductions: Participants were welcomed by the host and Meeting Chair, Dr. David Sands, Montana State University (MSU), Plant Science and Plant Pathology; Dr. Charles Boyer, Dean of MSU College of Agriculture; Dr. Allison Harmon, Dean of MSU College of Health and Human Services; and Dr. Tammy Bray, Dean, College of Public Health and Human Sciences, Oregon State University (Administrative Advisor).

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.

Dr. Juan Andrade was elected by the members present to be the chair of the next annual meeting. He will coordinate next year’s W3002 meeting in Urbana, Illinois, targeted for late May, 2016. Dr. Brian Lindshield, Kansas State University, was selected to be secretary.

Campus Tour of the Montana State University Plant Growth Center, Ag BioScience Building and BioChemistry Building (Dr. David Sands, Dr. Chaofu Lu, Dr. Edward Dratz).

Each W3002 Investigator attending the meeting provided an oral progress report in the following order:

1) ‘Diet, Gut Biome, and Bone,’ Connie Weaver, Purdue University

2) ‘Vitamin E Pharmacokinetics in Metabolic Syndrome Adults,’ Rich Bruno, Ohio State University

3) ‘Dietary Factors that Mitigate Inflammation and Cancer,’ Emily Ho, Oregon State University

4) ‘Role of Astaxanthin in the Prevention of Liver Fibrosis,’ Ji-Young Lee, University of Connecticut

5) ‘Contribution of Conjugated Linoleic Acid in Adipocyte, Muscle and Bone Metabolisms,’ Yeonhwa Park, University of Massachusetts

6) ‘Sleep Deprivation Reduces Energy Expenditure,’ Jennifer Teske, University of Arizona

7) ‘Protein Quality and Micronutrient Bioavailability of Extruded Corn, Soy, Sorghum and Cowpea Fortified-blended Foods,’ Brian Lindshield, Kansas State University

8) ‘Technologies to Assess Nutrient Status In and Deliver Micronutrients to Low-income Populations,’ Juan Andrade, University of Illinois

9) ‘Micronutrient Studies in Ethiopia,’ Barbara Stoecker, Oklahoma State University

10) ‘The Four Horseman and The Breeder’s Dilemma,’ David Sands, Montana State University

Discussion

Overall - Discussion regarding more connections between trace element intake and diseases.

Dr. Sands - The soil source of these trace elements is important to consider.

All - Ideas on methods on how to use mice models.

Regarding fortified food blends and international nutrition work, there was discussion on standards and regulations that place constraints on nutritional advancement.

Cultural models – for international work, domestic development and Native American nutrition

Dr. Dingbo Daniel Lin, Department of Nutritional Sciences, Oklahoma State University (New-W3002 participant), gave a presentation on ‘Overnutrition and BCO2 in the Development of Type 2 Diabetes.’

Interspersed within the W3002 presentations, the following invited speakers from Montana State University gave brief reports on related research to W3002:

1) ‘Microbiome’: Dr. Seth Walk, Dr. Carl Yeoman, Pilar Manrique

2) ‘Plasmid Curing’: undergraduate students Amanda Leckband and Andrew Helming

3) ‘Nutrition Research on US Reservations’: Dr. Holly Hunts, Dr. Edward Dratz

4) ‘Low Glycemic Potatoes’: Dr. Alice Pilgeram

The meeting adjourned on August 6, 2015, 3:00 p.m.

Accomplishments

The strength of this group can be indicated by their expressed interest in forming collaborative units based on common interests. We have numerous examples of suggestions for collaborations. Nutritional research, especially when combined with plant breeding can have a great impact on overall health and wellness of a population. The process of improving diets involves defining nutritional components, developing assays, creating public awareness of the resulting nutritionally improved food products, and finally incorporation into available products for consumers. Many of these collaborations were centered on eventual production of medically driven niche market foods for diabetics, celiac disease sufferers and similar groups. 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. University of Arizona (Jennifer Teske). Short-term Outcomes: i) adopted technology to quantify components of total energy expenditure; ii) Presentation skills training for oral and posters for 1 doctoral student, 2 Master’s degree students, 4 undergraduates, and 2 high school students; iii) Students received 4 awards based on research. Outputs: i) One oral presentation on research presented by undergraduate student at an International Scientific conference; ii) Six poster presentations on research presented by doctoral (2 posters), high school (3 posters), Master’s degree (1 poster) students; iii) Research opportunities provided for 1 doctoral student, 2 Master’s degree students, 4 undergraduates, and 2 high school students. Most activities were associated with the use of the integrated equipment high-resolution indirect calorimetry, EEG/EMG recordings and infrared beam-break activity sensors to determine metabolic rate during rest, rapid eye movement sleep, non-rapid eye movement sleep, and high and low intensity physical activity. Milestones associated with project: to quantify energy expenditure due to low and high intensity physical activity. 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 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). I) Obesity is closely tied to hyperlipidemia, chronic inflammation, and insulin resistance. To gain insight into the effect of polyphenol-rich blackcurrant extract (BCE) on obesity-associated abnormalities, C57BL/6J mice a high fat/high cholesterol diet supplemented with 0.1% BCE for 12 wk. Plasma total cholesterol and glucose, liver steatosis, macrophage infiltration in adipose tissue were all significantly lower with BCE supplementation compared with control. Ex vivo analysis of splenocytes isolated from control and BCE-fed mice showed that upon lipopolysaccharide stimulation, the expression of tumor necrosis factor ? and interleukin 1 ? were significantly reduced in the BCE group. OUTCOME: BCE supplementation may be beneficial to prevent metabolic disturbance and chronic inflammation in obesity. II) We explored if the post-transcriptional regulation of LDL receptor (LDLR) by BCE involves proprotein convertase subtilisin/kexin type 9 (PCSK9) compared with chockberry extract (CBE). PCSK9 mRNA and protein were drastically increased, while LDLR was markedly induced, by BCE and CBE. However, when Caco-2 cells were treated with rapamycin, a mechanistic target of rapamycin C1 inhibitor, together with BCE or CBE, rapamycin partly abolished the induction of LDLR protein by BCE, but with no effect on CBE-induced LDLR protein. Importantly, BCE significantly increased transintestinal cholesterol efflux of LDL-derived cholesterol compared to control. OUTCOME: BCE induces TICE via the post-transcriptional induction of LDLR expression partly by mTORC1-dependent, but PCSK9-independent, pathway in the intestine. III) Non-alcoholic steatohepatitis (NASH) is a subset of non-alcoholic fatty liver disease, the most common chronic liver disease in the U.S. Fibrosis, a feature of NASH, results from the dysregulation of fibrogenesis in hepatic stellate cells (HSCs). We investigated whether astaxanthin (ASTX), a xanthophyll carotenoid, can inhibit HSC activation. In LX-2 cells, accumulation of ROS induced by tert-butyl hydrogen peroxide and TGF?1, a key fibrogenic cytokine, was abolished by ASTX. ASTX significantly decreased TGF?1-induced ?-smooth muscle actin (?-SMA) and procollagen type 1, alpha 1 (Col1A1) mRNA as well as ?-SMA protein levels. Knockdown of Smad3 revealed Smad3 is required for ASTX to block TGF?1-induced fibrogenic gene expression. ASTX attenuated TGF?1-induced Smad3 phosphorylation and nuclear translocation with a concomitant inhibition of Smad3, Smad7, TGF? receptor I (T?RI), and T?RII expression. The inhibitory effect of ASTX on HSC activation was confirmed in primary HSCs as evidenced by decreased mRNA and protein levels of ?-SMA during HSC activation. Taken together, ASTX exerted anti-fibrogenic effects by blocking TGF?1-signaling and resulting activation of Smad3 in HSCs. University of Illinois, Urbana-Champaign (Juan Andrade). Our research focuses on using low-cost technologies or approaches to improve nutrition in developing countries. The following research themes have been pursued and built upon: 1) development of an immunological base photonic crystal biosensor for the assessment of iron deficiency anemia; 2) development of lipid based dietary supplements to treat acute malnutrition. OUTPUTS: I) Biosensor: a proof-of-concept platform using photonic crystal technology for detection and quantification of soluble transferrin receptor was developed. The biosensor combines extraction using magnetic properties of iron oxide nanoparticles with detection on a photonic crystal platform. Bias of quantification of control serum in biosensor is similar to that from commercial ELISAs. II) Lipid based supplements: a lipid nutrition supplement (LNS) for the prevention and treatment of acute malnutrition without edema was formulated. The product was evaluated in terms of stability and sensory acceptability with subjects in India. The Mansinhbhai Institute of Dairy & Food Technology and Dudhsagar Dairy are major partners for implementation. OUTCOMES: I) Biosensor (knowledge): the isolation of biomarkers of interest was improved using the magnetic properties of iron oxide nanoparticles. Non-specific binding was reduced due to isolation of antigens before detection. II) Lipid based supplement (knowledge): the formulation of LNS for India can lead to cost savings as production can be accomplished in India using local staples. Kansas State University (Brian Lindshield). We have focused on the potential of sorghum and cowpeas instead of corn and soybeans to produce micronutrient fortified-blended foods (FBFs) that are commonly used for food aid. Two white and one red blend of sorghum-cowpea with SPI or WPC, white sorghum-soy, extruded corn-soy blend, CSB+ (current USAID FBF), and AIN-93G were fed ad libitum to 21-23 day old male weanling Sprague-Dawley rats (n=10) for 4 weeks, while measuring food intake and weights. Blend nutritional contents, anthropometric, and micronutrient data were analyzed post hoc. The CSB+ had significantly lower weight gain, caloric efficiency, protein efficiency, length, and food intake. Compared with AIN-93G and white sorghum-cowpea with WPC, the SPI FBF group had significantly lower caloric efficiency. There were no significant differences between WPC FBFs groups. University of Maine (Dorothy Klimis-Zacas). The Metabolic syndrome (MetS) is a major public health problem in the United States. The MetS is characterized by the concurrent presence of obesity, dyslipidemia, insulin resistance, glucose intolerance and hypertension, and other associated metabolic abnormalities. Finding economic and effective ways to prevent and reverse the MetS is extremely important for public health and the health care system, especially considering that its prevalence is dramatically increasing among the US population. Wild blueberries are among the commercially available fruits and vegetables that contain the highest levels of antioxidant polyphenols, mostly anthocyanins (ACNs). Past research in our laboratory has documented that wild blueberries beneficially affect the cardiovascular system and regulate vasomotor tone and arterial metabolism in animal models of normal endothelial function but also dysfunction, correct/normalize the dyslipidemia associated with the MetS and attenuate the inflammatory state. Preliminary results on the effects of wild blueberry consumption on plasma markers and gene expression related to glucose metabolism in the obese Zucker rat (OZR), an experimental model of metabolic syndrome and their lean littermate (LZR), documented that consumption of an 8% enriched wild blueberry diet (WB) for 8 weeks resulted in significant reduction of plasma glycated hemoglobin HbA1c, RBP4, and resistin concentrations in OZR. Additionally, following WB consumption, resistin expression was significantly down regulated in the liver of both OZR and LZR while RBP4 expression was significantly down regulated in the adipose tissue of both OZR and LZR. Thus, wild blueberry consumption normalizes some markers related to glucose metabolism and gene expression in the OZR, but has no effect on fasting blood glucose or insulin concentration. OUTPUTS: Results from the period covered in this report were disseminated at the Experimental Biology Meetings, April 2015, through published articles, presented at invited lectures and through the media (newspaper articles, blogs and interviews). University of Massachusetts (Yeonhwa Park). We have studied various bioactive food components for their bioavailability and metabolism. Consumption of lipophilic bioactive agents (such as fucoxanthin, nobiletin, oil-soluble vitamins, conjugated linoleic acid) may reduce the incidences of certain chronic diseases. However, their utilization in foods is currently limited because of their poor water-solubility, high melting point, low bioavailability, and chemical instability. We have examined the impact of a number of different factors on the encapsulation, protection, and release of lipophilic bioactive agents, including carrier oil type, particle size, and emulsifier type. In addition, we also investigated the role of nobiletin and its colonic metabolites in a colitis-associated colon carcinogenesis model. This information has enabled us to identify mechanisms of lipophilic bioactives as well as to develop optimized delivery systems that can be used to incorporate lipophilic bioactive agents into foods and increase their oral bioavailability. Montana State Univeristy (David Sands). The research at Montana State University has involved coordination between four laboratories, and has emphasized development of high omega-3 animal feeds, and microgreens made with Camelina sativa, and low glycemic lines of potatoes. Camelina sativa is a new omega-3 oil and protein crop that in many ways rivals soy. It is fed to chickens for production of high omega-3 eggs 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-round source of fresh greens. Steps are being made to introduce them into the restaurant industry through a kitchen growth chamber company. Aiming to develop a more diabetic-friendly potato, seven lines of potatoes that are lower in glycemic index are being analyzed for human consumption. 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. PrOatina, a high-protein, low glycemic hull-less oat is being pearled to remove bitter saponins and increase similarity to rice. OUTPUTS: I) Advanced testing of low glycemic potatoes for introduction into the retail market. II) Development of microgreens of Camelina sativa for the home and restaurant market. III) Development of tests for plasmid curing agents in crop plants. IV) 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. V) Development of a less-bitter high protein oat through a saponin-removal pearling process. OUTCOMES: I) Identify low glycemic potato lines for possible value in obesity mitigation. II) Establish Camelina microgreens as a source of omega-3 fatty acids. Lower glucosinolate lines are now available. III) Establish a Sentinel Seed System to enable gardeners and subsistence farmers to know what elements are deficient in their soil. University of Nebraska, Lincoln (Janos Zempleni). We have continued our research of bioactive food compounds and their effects on human health along the following lines. (1) We have discovered synthetic and natural inhibitors of holocarboxylase synthetase (HLCS). The studies of natural HLCS inhibitors focused on resveratrol metabolites in grape products. Using these inhibitors we demonstrated that resveratrol metabolites cause a decrease in body fat mass in Drosophila melanogaster mutants that are genetically predisposed to high body fat mass, but by inhibition of insulin receptor signaling. (2) 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. (3) We have continued to shift the focus of our research program away from biotin metabolism and HLCS catalysis to a novel class of bioactive food compounds that we have discovered, i.e., microRNAs packaged in extracellular vesicles (EVs) in cow’s milk and chicken eggs. Briefly we have demonstrated that 1) human intestinal and vascular endothelial cells transport EVs and their microRNA cargo by endocytosis, 2) humans absorb biologically effective amounts of milk and egg microRNAs, 3) dietary microRNAs are delivered to peripheral tissues where they affect human gene expression, 4) endogenous synthesis of microRNAs does not compensate for dietary microRNA depletion, and 5) dietary microRNA depletion elicits phenotypes such as low fecundity, increased purine metabolism, and low body fat mass in mice. These studies of the phenotypes of dietary microRNA status are an important milestone guiding our ongoing studies of the mechanisms that elicit these phenotypes. OUTCOMES: We have discovered a novel class of bioactive compounds in foods, i.e., microRNAs. This research has major implications for the U.S. dairy and egg industry, and the way we assess the nutritional value of foods. Ohio State University (Richard Bruno). ?-Tocopherol (?-T) protects against nonalcoholic steatohepatitis (NASH), but its dietary intakes are poor with >22% of Americans having suboptimal ?-T status. We therefore tested the hypothesis that MetS adults, a population at high risk for NASH, would have lower ?-T bioavailability that could be improved by fat-containing dairy milk. A randomized, crossover, double-blind study was conducted in healthy and MetS adults (n = 10/group) who ingested encapsulated deuterium-labeled (d6)-RRR-?-T (15 mg) with 240 mL of non-fat (0.2 g fat), reduced-fat (4.8 g fat), or whole (7.9 g fat) milk prior to blood collection at regular intervals for 72 h. Compared to healthy participants, those with MetS had lower (P<0.05) baseline plasma ?-T (?mol/mmol lipid) and greater oxidized LDL (oxLDL), IL-6, IL-10, and C-reactive protein (CRP). Regardless of health status, d6-?-T bioavailability was unaffected by increasing amounts of dairy fat provided by milk beverages, but MetS participants had lower estimated d6-?-T absorption compared with healthy participants. They also had lower plasma d6-?-T AUC0-72 h and maximal concentrations (Cmax), and slower rates of plasma disappearance, but similar times to Cmax. MetS participants had lower d6-?-T AUC0-t final in lipoprotein fractions (chylomicron, VLDL, LDL, HDL), and %d6-?-T AUC0-t final in both the chylomicron and VLDL fractions were inversely correlated with oxLDL, IL-10, IL-6, and CRP. At dietary intakes equivalent to the Recommended Daily Allowance, ?-T bioavailability is unaffected by dairy fat quantity, but is lower in MetS adults, likely due to greater inflammation and oxidative stress that limits small intestinal ?-T absorption and/or impairs hepatic ?-T trafficking. These findings support higher dietary ?-T requirements for MetS adults. Oklahoma State University (Barbara Stoecker). Our laboratory focuses on the effects of micronutrient deficiencies on human health. We have investigated the effects of zinc supplementation in women in the context of the high prevalence of dietary zinc deficiency in Ethiopia. Data obtained from one trial included information on increased DNA breaks in women not supplemented with Zn compared to women who received the supplement. In another trial, zinc supplementation decreased abundance of mRNA for ZIP4 and ZIP8 in peripheral blood mononuclear cells of adult women. We have provided serum samples from women with very limited zinc intakes to colleagues in the W3002 multistate project to support metabolomics studies related to the roles of zinc in human health. We also are conducting a randomized control trial of vitamin D supplementation of lactating women in rural Ethiopia. Skin color is being measured as well as UVB light exposure. Both breast milk and blood samples have been collected. Dietary analysis indicates a very low intake of vitamin D. The vitamin D status biomarker in serum, 25(OH) vitamin D, and vitamin D in breast milk are being measured. Morbidity and motor milestones of infants are being measured from birth throughout their first year of life. Field work for this study will be completed in early 2016. 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, and inflammatory markers of pre-diabetic individuals. Our findings indicate that chronic consumption of freeze-dried mango by pre-diabetic individuals does not negatively affect blood glucose and inflammatory markers. We also investigated the effects of freeze-dried mango on gut microbiome in mice fed high fat diet. Mango supplementation prevented the loss of Bifidobacteria and Akkermansia due to high fat feeding. Mango exerts this effect in a dose-dependent manner and is statistically similar to mice fed normal diet. Oregon State University (Emily Ho). Zinc and chronic disease: Recently we have found that zinc status is compromised with age. Zinc deficiency causes immune cells to be sensitized to inflammation and also occurs in older animals and humans. 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. Supplemental SFN did show benefits in decreasing cell proliferation markers and histone deacetylase 3 expression in breast tissue in women previously diagnosed with breast cancer. GOALS: I) Test the effects of zinc status on the gut microbiota, epigenetics, oxidative stress, inflammation, DNA damage and cancer susceptibility in rodent models and humans. II) Understand the determinants of bioavailability of phytochemicals derived from cruciferous vegetables. III) Test the ability of sulforaphane supplementation from various sources (supplement vs whole food) to reduce the incidence of prostate and breast cancer via epigenetic modifications. Oregon State University (Norman Hord). This report summarizes progress to address potential health benefits of plant food-derived dietary nitrate. Since nitrate and nitrite have been associated with risk of methemoglobinemia in newborns and gastrointestinal cancer risk in adults when consumed in processed meats, nitrate concentrations in drinking water are regulated by the U.S. EPA. With regard to dietary nitrate intake, vegetable and fruit intake constitute the source of 80% of nitrate in the diet and these food groups are positively associated in epidemiologic studies with decreased risk of cardiovascular disease and low bone mass. Our projects work across the spectrum from food-based analysis to zebrafish and rodent model systems to assess dietary exposures, potential toxicities and potential health benefits, respectively. Each represents preliminary data being prepared for publication. Project 1: We have assessed nitrate and nitrite concentrations using foods and mixed dishes representing one day of intakes from traditional food patterns, rather than using limited food database information available, from a variety of cultural patterns in order to estimate actual intake levels. Outcome 1: Our data indicate that these cultural patterns contained diverse nitrate concentrations (range ~350 to 100 mg/day nitrate), represented as a rank order in nitrate concentration from one day food intake patterns as follows: Mediterranean>Chinese>Japanese>American=Indian. This rank order is consistent with the hypothesis that nitrate intakes may form an inverse ecological association with cardiovascular disease rates in these countries; in clinical studies, the highest concentrations measured in these dietary patterns have been demonstrated to produce cardioprotective benefits. Project 2: We have interrogated the effect of dietary nitrate in pilot studies in rodents to determine whether it reduces ovariectomy-induced bone loss in female rats or improves glucose tolerance in male mice fed a Western-type dietary pattern. Outcome 2: In both of these rodent studies, we found that dietary nitrate does not reduce bone loss or improve glucose tolerance in the rodent models examined. Project 3: The objective of this project funded by the OSU Environmental Health Sciences Center, was to identify a potential role for dietary nitrate-derived NO in fetal cardiovascular development and to investigate whether supplementation with dietary nitrate (10, 50, 100, 250, 500, and 1000 µM/L) prevents or reduces occurrence of congenital cardiovascular defects (CCVD) in zebrafish embryos. These concentrations are selected on the basis of published literature on plasma nitrate levels in normal and pregnant conditions, as well as on the basis of estimates of per capita daily dietary nitrate intake for the average US population (75 mg, ? 1 mM). Outcome 3: Our data indicate that only dietary nitrate concentrations of 1000 µM/L were associated with occurrence of CCVD. These data indicate minimal risk of CCVD due to dietary nitrate concentrations typically consumed in U.S. populations. Purdue University (Connie Weaver). Prebiotics enhanced mineral absorption in animal models and humans and bone parameters in animal models. We extended our studies on the effect of soluble corn fiber (SCF) on enhancing calcium absorption in adolescents to study the effect on bone calcium retention in postmenopausal women. We used our novel approach of urinary excretion of the rare isotope, 41Ca, from pre-labeled bone to test the dose responsive effect of feeding 0, 10 and 20 g SCF/d. SCF increased (p<0.01) bone calcium retention in a dose responsive manner.

Publications

Ahluwalia, A., Gladwin, M., Coleman, G.D., Hord, N.G., Howard, G., Kim-Shapiro, D., Lajous, M., Larsen, F., Lefer, D.J., McClure, L.A., Nolan, B.T., Pluta, R., Schechter, A., Wang, C.-Y., Ward, M.H., and Harman, J.L. (2015) Dietary nitrate and the epidemiology of cardiovascular disease: Report from a National Heart, Lung, and Blood Institute Workshop, Circulation (submitted for publication). Anderson RA, Zhan Z, Luo R, Mu X, Guo X, Liu Y, Guo Q, Zhou J, Kong J, Cui B, Davis PA, Stoecker BJ. Cinnamon extract lowers glucose, insulin and cholesterol in people with elevated serum glucose. Journal of Traditional and Complementary Medicine (In Press). Atwell LL, Hsu A, Wong CP, Stevens JF, Bella D, Yu TW, Pereira CB, Löhr CV, Christensen JM, Dashwood RH, Williams DE, Shannon J, Ho E. (2015) Absorption and chemopreventive targets of sulforaphane in humans following consumption of broccoli sprouts or a myrosinase-treated broccoli sprout extract. Mol Nutr Food Res. doi: 10.1002/mnfr.201400674. Baier SR, Nguyen C, Xie F, Wood JR, Zempleni J. MicroRNAs are absorbed in biologically meaningful amounts from nutritionally relevant doses of cow’s milk and affect gene expression in peripheral blood mononuclear cells, HEK-293 kidney cell cultures, and mouse livers. J Nutr 144:1495-1500, 2014 Baier SR, Nguyen C, Xie F, Wood JR, Zempleni J. Reply to Witwer: Mammals absorb microRNAs from milk. J Nutr 144:1881, 2014 [Letter to the editor] Bailey, L. B., Stover, P., McNulty, H., Fenech, M., Gregory, J., James Mills, J., Pfeiffer, C. M., Fazili, Z., Zhang, M., Ueland, P., Molloy, A., Caudill, M. A., Shane, B., Berry, R. J., Bailey, R., Raghavan, R. and Raiten, D. (2015) Biomarkers of Nutrition for Development (BOND) - Folate Review. J Nutr 145: 1636S-80S. Ballard KD, RS Bruno. (2015). Protective Role of Dairy and its Constituents on Vascular Endothelial Function Independent of Blood Pressure-Lowering Activities. Nutr Rev. 73(1):36-50. Beaver LM, Buchanan A, Sokolowski EI, Riscoe AN, Wong CP, Chang JH, Löhr CV, Williams DE, Dashwood RH, Ho E. (2014) Transcriptome analysis reveals a dynamic and differential transcriptional response to sulforaphane in normal and prostate cancer cells and suggests a role for SP1 in chemoprevention. Mol Nutr Food Res. 2014 Oct;58(10):2001-13. doi: 10.1002/mnfr.201400269. Epub 2014 Aug 5. Benn T, B. Kim, Y. Park, C. J. Wegner, E. Harness, T-G. Nam, D-O. Kim, J. S. Lee, J. Lee. Polyphenol-rich blackcurrant extract prevents inflammation in diet-induced obese mice. J Nutr Biochem 2014; 25:1019-25. Benn T, B. Kim, Y. Park, Y. Yang, T. X. Pham, C.S. Ku, E. Harness, C. Farruggia, E. Harness J. A. Smyth, J. Lee. Polyphenol-rich blackcurrant extract exerts hypolipidemic and hypoglyceridemic effects in mice fed a diet containing high fat and cholesterol. Br J Nutr 2015: 113:1697-1703. Bloedon T.K., S. Vendrame, J.C. Bolton, R.A. Lehnhardt, P. Riso and D. Klimis- Zacas, The effect of wild blueberry (V. angustifolium) consumption on oxidative stress, inflammation and DNA damage associated with exercise, Comparative Exercise Physiology, 11 (3), 173-81, 2015 Bogale A, Clarke SL, Fiddler J, Hambidge KM, Stoecker BJ. (2015) Zinc supplementation decreased ZIP4 and ZIP8 mRNA abundance in peripheral blood mononuclear cells of adult women. Nutrition and Metabolic Insights 8:7-14. doi:10.4137/NMI.S23233. Camara Teixeira D, Cordonier EL, Wijeratne SSK, Huebbe P, Jamin A, Jarecke S, Wiebe M, Zempleni J. A cell death assay for assessing the mitochondrial targeting of proteins (revision submitted) Carter, T. C., Pangilinan, F., Molloy, A. M., Fan, R., Wang, Y., Shane, B., Gibney, E. R., Midttun, O., Ueland, P. M, Cropp, C. D., Kim, Y., Wilson, A. F., Bailey-Wilson, J. E., Brody, L. C. and Mills, J. L. (2015) Common variants at putative regulatory sites of the tissue nonspecific alkaline phosphatase gene influence circulating pyridoxal 5’-phosphate concentration in healthy adults. J. Nutr. 145: 1386-93. Chiang K, Shu J, Zempleni J, Cui J. Dietary MicroRNA Database (DMD): an archive database and analytic tool for food-borne microRNAs. PLoS ONE (in press) Chongwatpol P, Rendina-Ruedy E, Stoecker BJ, Clarke SL, Lucas EA, Smith BJ. Implications of compromised zinc status on bone loss associated with chronic inflammation in C57BL/6 mice. J Inflam Res. 2015; 8: 117-28. Chung M-Y, E Mah, C Masterjohn, SK Noh, HJ Park, RM Clark, Y-K Park, J-Y Lee, RS Bruno. (2015). Green tea lowers hepatic COX-2 and prostaglandin E2 in rats with dietary fat-induced nonalcoholic steatohepatitis. J Med Food, 18(6):648-55. Cordonier EL*, Adjam R*, Camara Teixeira D, Onur S, Zbasnik R, Read PE, Döring F, Schlegel VL, Zempleni J. (*contributed equally). Resveratrol compounds inhibit human holocarboxylase synthetase and cause a lean phenotype in Drosophila melanogaster. J Nutr Biochem (in press) Deac, O. M., Mills, J. L., Shane, B., Midttun, O., Ueland, P. M., Brosnan, J. T., Brosnan, M. E., Laird, E., Gibney, E. R., Fan, R., Wang, Y., Brody, L. C. and Molloy, A. M. (2015) Tryptophan catabolism and vitamin B-6 status are affected by gender and lifestyle factors in healthy young adults. J. Nutr. 145: 701-7. Del Bo’ C, Martini D, Porrini M, Klimis-Zacas D, Riso P. Berries and oxidative stress markers: an overview of human intervention studies, Food and Function, 2015, DOI: 10.1039/c5fo00657k Del Bo’ C, Porrini M, Campolo J, Parolini M, Lanti C, Klimis-Zacas D and Riso P. Effect of a single blueberry (V. corymbosum) portion on markers of antioxidant defense and oxidative stress in healthy volunteers following cigarette smoking: a randomized-controlled study, Food and Function, In Press, 2015 Evans SF, Meister M, Peterson S, Perkins-Veazie P, Clarke SL, Payton M, Smith BJ, Lucas EA. Mango supplementation improves blood glucose in obese individuals. Nutrition and Metabolic Insights. 2014, 7:77-84. Fekadu Y, Mesfin A, Haile D, Stoecker, BJ. (2015) Factors associated with nutritional status of infants and young children in Somali Region, Ethiopia: a cross-sectional study. BMC Public Health 2015, 15:846. DOI: 10.1186/s12889-015-2190- Fenton, JI, Hord, NG, Ghosh, S and E Gurzell (2013) Long chain omega-3 fatty acid overexposure and immunomodulation. Prostaglandins Leukot Essent Fatty Acids. Sep 30. pii: S0952-3278(13)00192-0. doi: 10.1016/ j.plefa. 2013. 09.011. [Epub ahead of print] Guo Y, E Mah, RS Bruno. (2014) Quercetin Bioavailability Is Associated With Inadequate Plasma Vitamin C Status And Greater Plasma Endotoxin In Healthy Adults. Nutrition, 30(11-12):1279-1286. Guo Y, RS Bruno. (2014). Endogenous and exogenous mediators of quercetin bioavailability. (Review). Submitted, J Nutr Biochem, 26(3):201-210. Hohman EE, McCabe GP, Peacock M, Weaver CM. Validation of urinary calcium isotope excretion from bone for screening anabolic therapies for osteoporosis. Osteoporos Int 25:2471-2475, 2014. Hohman EE, Weaver CM. A grape-enriched diet increases bone calcium retention and cortical bone properties in ovariectomized rats. J Nutr 145:253-259, 2015. Houghton-Rahrig, L., Schutte, D., von Eye, A., Fenton, J.I., Given, B.A., & Hord, N.G. (2013). Exploration of a symptoms experience in persons with NAFLD. Nursing Outlook. Jul-Aug;61(4):242-251.e2. doi: 10.1016/j.outlook.2013.05.003. Howard KM, Jati Kusuma R, Baier SR, Friemel T, Markham L, Vanamala J, Zempleni J. Loss of miRNAs during processing and storage of cow’s (Bos taurus) milk. J Agr Food Chem 63:588-592, 2015 Jaryum KH, Okoye ZSC, Stoecker B. (2013) Copper content of staple seeds and grains grown in Kanam local government area, Nigeria. SpringerPlus 2:373-377. Joray ML, Yu TW, Ho E, Clarke SL, Stanga Z, Gebreegziabher T, Hambidge KM, Stoecker BJ. (2015) Zinc supplementation reduced DNA breaks in Ethiopian women. Nutr Res 35:49-55. Kang Y, Nian H, Rajendran P, Kim E, Dashwood WM, Pinto JT, Boardman LA, Thibodeau SN, Limburg PJ, Löhr CV, Bisson WH, Williams DE, Ho E, Dashwood RH. (2014) HDAC8 and STAT3 repress BMF gene activity on colon cancer cells. Cell Death Dis. 2014 Oct 16;5:e1476. doi: 10.1038/cddis.2014.422. Kim B, C. Farruggia, C. S. Ku, T. X. Pham, Y. Yang, C. J. Wegner, N. J. Farrell, E. Harness, Y. Park, S. I. Koo, J. Lee. Astaxanthin, a xanthophyll carotenoid, inhibited inflammation and fibrosis in the adipose and the liver of diet-induced obesity and liver fibrosis mice. J Nutr Biochem 2015; (In review). Kim Y. and Y. Park (2015) Conjugated Linoleic Acid (CLA) Stimulates Mitochondrial Biogenesis Signaling by the Upregulation of Peroxisome Proliferator-Activated Receptor- ? Coactivator 1? in C2C12 Cells, Lipids 50: 329-338. Kusuma JR, Friemel T, Zempleni J. Human vascular endothelial cells transport foreign exosomes from cow’s milk by endocytosis (revision submitted) Li Y, Malkaram SA, Zhou J, Zempleni J. Lysine biotinylation and methionine oxidation in the heat shock protein HSP60 synergize in the elimination of reactive oxygen species in human cell cultures. J Nutr Biochem 2014 Apr;25(4):475-82. Liu D, Zempleni J. Holocarboxylase synthetase interacts physically with the nuclear receptor corepressor, histone deacetylase 1, and a novel splicing variant of histone deacteylase 1 to repress repeats. Biochem J 461:477-486, 2014 Liu D, Zempleni J. Low activity of LSD1 elicits a pro-inflammatory gene expression profile in riboflavin-deficient human T lymphoma Jurkat cells. Genes Nutr 9:422, 2014 Liu D, Zempleni J. Transcriptional regulation of the albumin gene depends on the removal of histone methylation marks by the FAD-dependent monoamine oxidase LSD1 in HepG2 human hepatocarcinoma cells. J Nutr 144:997-1001, 2014 Mah E, R Pei, Y Guo, C Masterjohn, KD Ballard, BA Parker, AW Taylor, MG Traber, JS Volek, RS Bruno. (2014). Greater ?-tocopherol status during acute smoking abstinence with nicotine replacement therapy improved vascular endothelial function by decreasing 8-iso-15(S)-prostaglandin F2?. Exp Biol Med, 240(4):527-33. Mavanji V, Perez-Leighton CE, Kotz CM, Billington CJ, Parthasarathy S, Sinton CM, and Teske, JA. Promotion of wakefulness and energy expenditure by orexin A in the ventrolateral preoptic area. Sleep. (in press). Epub 2015 April 8. McKenney ML, Territo PR, Salavati Al, Houshmand S, Persohn S, Liang Y, SturekJM, Alloosh M, Moe SM, Weaver CM, Alavi A, Sturek, M. 18F-NaF positron emission tomography imaging of early coronary artery calcification. J Am Coll Cardiology Accepted, 2015. Mills, J. L., Fan, R., Brody, L. C., Liu, A., Ueland, P. M., Wang, Y., Kirke, P. N., Shane, B. and Molloy, A. M. (2014) Maternal choline concentrations during pregnancy and choline-related genetic variants as risk factors for neural tube defects. Am J Clin Nutr 100: 1069-74. Nakatsu CH, Weaver CM, Martin BR, Clavijo A, Barnes S. Fecal bacterial community changes associated with isoflavone metabolites in postmenopausal women after soy bar consumption. PONE 9:e108924, 2014. Nixon JP, Mavanji V, Butterick TA, Billington CJ, Kotz CM and Teske JA. Sleep disorders, aging and obesity: the role of orexin. Ageing Research Reviews. 20C: 63-73. 2015. Epub 2014 Nov 22. Noriega, K.E., Lindshield, B.L. Is the Inclusion of Animal Source Foods in Fortified Blended Foods Justified? Nutrients. 6(9), 3516-3535, 2014. doi:10.3390/nu6093516 Ozaki, M., Molloy, A. M., Mills, J. L., Fan, R., Wang, Y., Gibney, E. R., Shane, B., Brody, L. C. and Parle-McDermott, A. (2015) The Dihydrofolate Reductase 19bp polymorphism has no significant impact on biomarkers of folate status in healthy young adults, irrespective of folic acid intake. J. Nutr. doi: 10.3945/jn.115.216101 Palacios C, Martin BR, McCabe GP, McCabe L, Peacock M, Weaver CM. Dietary calcium requirements do not differ between Mexican American boys and girls. J Nutr 144:1167-1173, 2014. Pangilinan, F., Molloy, A. M., Mills, J. L., Troendle, J. F., Parle-McDermott, A., Kay, D. M., Browne, M. L., McGrath, E. C., Abaan, H. O., Sutton, M., Kirke, P. N., Caggana, M., Shane, B., Scott, J. M. and Brody, L. C. (2014) Replication and Exploratory Analysis of 24 Candidate Risk Polymorphisms for Neural Tube Defects. BMC Medical Genetics Oct 8;15:102. doi: 10.1186/s12881-014-0102-9. Park CY, Lee WH, Fleet JC, Allen MR, McCabe GP, Walsh DM, Weaver CM. Calcium and vitamin D intake maintained from pre-ovariectomy independently affect calcium metabolism and bone properties in Sprague Dawley rats. J Nutr 25:1905-15, 2014. Pawlowski J, Martin B, McCabe G, Ferruzzi M, Weaver, C. Plum and soy aglycon extracts superior at increasing bone calcium retention in ovariectomized Sprague Dawley rats. J Ag Food Chem. 62:6108-14, 2014. Pawlowski J, Martin B, McCabe G, McCabe L, Jackson G, Peacock M, Barnes S, Weaver CM. Impact of equol producing capacity and soy isoflavone profiles of supplements on bone calcium retention in postmenopausal women: a partially randomized crossover trial. AJCN Accepted. Pei R, E Mah, SW Leonard, MG Traber, RS Bruno. (2015). ?-Tocopherol supplementation reduces 5-nitro-?-tocopherol accumulation by decreasing ?-tocopherol in young adult smokers. Free Radic Res, 49(9):1114-21. Pei R, M Yu, RS Bruno, B Bolling. (2015). Phenolic and tocopherol content of autumn olive (Elaeagnus umbellate) berries. J Func Foods, 16:305-314. Peterson, RD, Chen, W, Cunningham, BT, Andrade, JE. 2015. Enhanced sandwich immunoassay using antibody-functionalized magnetic iron-oxide nanoparticles for extraction and detection of soluble transferrin receptor on a photonic crystal biosensor. Biosensors and Bioelectronics. 74: 815-822 Rendina-Ruedy E, Graef JL, Davis MR, Hembree KD, Gimble JM, Clarke SL, Lucas EA, Smith BJ. Strain differences in the attenuation of bone accrual in a young growing mouse model of insulin resistance. J Bone Miner Metab. 2015 Jun 10. Rendina-Ruedy E, Hembree K, Sasaki A, Davis M, Lighfoot SA, Clarke SL, Lucas EA, Smith BJ. A comparative study of the metabolic and skeletal response of C57BL/6J and C57BL/6N mice in a diet-induced model of type 2 diabetes. J Nutr Metab (in press) Salvia-Trujillo L, Q. Sun, B.-H. Um, Y. Park, and D. J. McClements (2015) In Vitro and In Vivo Study of Fucoxanthin Bioavailability From Nanoemulsion-Based Delivery Systems: Impact of Lipid Carrier type, J. Func. Foods 17: 293-304. Schafer AL, Weaver CM, Black DM Wheeler AL, Chang H, Szefc GV, Steward L, Rogers SJ, Carter JT, Posselt AM, Shoback DM, Sellmeyer DE. Intestinal calcium absorption decreases dramatically after gastric bypass surgery despite optimization of vitamin D status. J Bone Miner Res 30:1377-1385, 2015. Schwartz, K, Chang HT, Nikolai, M, Pernicone, J, Rhee, S, Olson, K, Kurniali PC, Hord, NG; Noel, M. (2015) Treatment of glioma patients with ketogenic diets: report of two cases treated with an IRB-approved energy-restricted ketogenic diet protocol and review of the literature. Cancer & Metabolism 3:3, DOI: 10.1186/s40170-015-0129-1 Shu J, Chiang K, Zempleni J, Cui J. Computational characterization of exogenous microRNAs that can be transferred into human circulation. PLoS ONE (in press) Sittiwong W, Cordonier EL, Zempleni J, Dussault P. ?-Keto and ?-hydroxyphosphonate analogs of biotin-5'-AMP are inhibitors of holocarboxylase synthetase. Bioorg Med Chem Lett 15:5568-5571, 2014 Teske JA, Perez-Leighton C, Billington CJ and Kotz CM. Methodological considerations for measuring spontaneous physical activity in rodents. American Journal of Physiology, Regulatory, Integrative and Comparative Physiology. 306(10): R714-21. 2014. Epub 2014 Mar 5. Torres, PC, Flores, F, Andrade, JE. 2015. Development of an extruded micronutrient delivery vehicle for the fortification of nixtamalized maize with chelated iron at the point of wet milling. Intl. J. Food Sci. Tech. 95(2):393-400. Vendrame S and D Klimis-Zacas, The anti-inflammatory effect of anthocyanins via the modulation of Nf-kB and MAPKs signaling cascades: A review, Nutrition Reviews, 73 (6), 348-58, 2015 Vendrame S, A. Daugherty, A.S. Kristo, D. Klimis-Zacas, Wild blueberry-enriched diet (Vaccinium angustifolium) improves dyslipidemia and modulates gene expression related to lipid metabolism in obese Zucker rats, British Journal of Nutrition, 111:194- 200, 2014 Vendrame S, A. S. Kristo, D. A. Schuschke, D. Klimis-Zacas, Wild blueberry consumption affects arterial vascular function in the obese Zucker rat, Applied Physiology, Nutrition and Metabolism, 39;255-61, 2014 Vendrame S., Zhao A., Merrow T., Klimis-Zacas D., The effects of wild blueberry consumption on glucose metabolism in the obese Zucker rat, a model of metabolic syndrome. Journal of Medicinal Food, DOI:10.1089/jmf.2014.0065, 2014 Volk BM, LJ Kunces, DJ Freidenreich, BR Kupchak, C Saenz, JC Artistizabal, ML Fernandez, RS Bruno, CM Maresh, WJ Kraemer, SD Phinney, JS Volek. (2014). Effects of step-wise increases in dietary carbohydrate on circulating saturated fatty acids and palmitoleic acid in adults with metabolic syndrome. PLoS One. 21;9(11):e113605. Watson GW, Wickramasekara S, Palomera-Sanchez Z, Black C, Maier CS, Williams DE, Dashwood RH, Ho E. (2015) SUV39H1/H3K9me3 attenuates sulforaphane-induced apoptotic signaling in PC3 prostate cancer cells. Oncogenesis. doi: 10.1038/oncsis.2014.47. Whisner CM, Martin BR, Nakatsu CH, McCabe GP, McCabe LD, Peacock M, Weaver CM. Soluble maize fibre affects short-term calcium absorption in adolescent boys and girls: a randomized controlled trial using dual stable isotopic tracers. Br J Nutr 112:446-456, 2014. Wolf T, Baier SR, Zempleni J. The Intestinal Transport of bovine milk exosomes is mediated by endocytosis in human colon carcinoma Caco-2 cells and rat small intestinal IEC-6 cells. J Nutr (in press) Wong CP, Rinaldi NA, Ho E. (2015) Zinc deficiency enhanced inflammatory response by increasing immune cell activation and inducing IL6 promoter demethylation. Mol Nutr Food Res. 2015 Feb 5doi: 10.1002/mnfr.20140076 Wu X, Song M, Wang M, Zheng J, Gao Z, Xu F, Zhang G, Xiao H, Chemopreventive effects of nobiletin and its colonic metabolites on colon carcinogenesis, Mol. Nutr. Food Res. (Accepted) Yang Y, B. Kim, Y. Park, S. I. Koo, J. Lee. Astaxanthin prevents transforming growth factor b1-induced pro-fibrogenic gene expression by inhibiting Smad3 activation in hepatic stellate cells. Biochimica Biophysica Acta 2015; 1850:178-185. Yang Y, M. Bae, B. Kim, Y. Park, S. I. Koo, J. Lee. Astaxanthin prevents and reverses the activation of mouse primary hepatic stellate cells. J Nutr Biochem 2015; (In review).

Impact Statements

The following grants were obtained: I) Teske, J. Hamilton Company, $1000 (5/2015-8/2015); II) Teske, J. University of Arizona Office of Global Initiatives, $1000 (1/15-11/15); III) Teske, J. University of Arizona-College of Agriculture and Life Sciences, $20,200 (6/15 ? 5/16); Teske, J. University of Arizona Office of Global Initiatives, $700 (11/14-3/15). University of Arizona, Jennifer Teske
The following student funds were obtained: I) Coborn, Univ. Arizona- Dept. Nutritional Sciences DeBell Research Enhancement, $2500, (6/2015-5/2016). II) DePorter, Univ. of Arizona- Office of the Vice President for Student Affairs and Enrollment Management, $500 (6/2015-7/2015); III) DePorter, Univ. of Arizona- Dept. Nutritional Sciences, $250 (6/2015-7/2015). University of Arizona, Jennifer Teske
Low maternal choline intake and blood concentration were thought to be risk factors for having a child with a neural tube defect (NTD). We examined the relationship between NTDs, choline status, and genetic polymorphisms reported to influence de novo choline synthesis to investigate claims that taking choline periconceptionally could reduce NTD rates. University of California, Berkeley, Barry Shane
Our results indicate that maternal betaine and choline concentrations are not strongly associated with NTD risk. An association between PEMT rs7946 and NTDs requires confirmation. University of California, Berkeley, Barry Shane
We demonstrate that blackcurrant and astaxanthin consumption may provide health benefits against obesity-associated inflammation, liver fibrosis and other metabolic dysfunctions. University of Connecticut, Ji-Young Lee
Low-cost biosensing technologies can support identification of populations suffering from micronutrient deficiencies, facilitating their care and reducing wasteful use of resources. A lipid nutrition with enhanced sensory characteristics can improve children and mother nutrition at Anganwadi Centers in India. The use of the distribution logistics of the Indian dairy cooperative system can increase effectiveness of nutrition delivery in rural areas. University of Illinois, Juan Andrade
Compared with new FBFs, CSB+ intake resulted in growth retardation, further research is needed to determine the reason for result. Outcomes in new FBFs were similar, and comparable to AIN-93G. Our results suggest that SPI FBFs may be of inferior protein quality compared to WPC FBFs, but that newly developed FBFs may be superior to CSB+. Kansas State University, Brian Lindshield
Regular wild blueberry consumption normalizes markers related to glucose metabolism and gene expression that may influence the insulin resistant state associated with the Metabolic syndrome. The above research project impacted graduate, undergraduate students and visiting scientists, acquiring skills and knowledge on the area of food bioactives and their effects on health as well as the Blueberry Industry and other commodity groups. University of Maine, Dorothy Klimis-Zacas
Lipophilic nutraceuticals have been implied to have major health benefits. However they are difficult to add to the diet due to their low-water solubility, poor chemical stability and low bioaccessibility. Development of delivery system for lipophilic bioactives in food can increase the application of these bioactives into foods. University of Massachusetts, Yeonhwa Park
Plant breeding is generally focused on yield and agronomic characteristics. Protein content, essential amino acid balance and critical omega-3:6 ratios are not of primary concern. This research is significant because it demonstrates that such nutritional traits are within the purview of plant breeding and that they can lead to more nutritious foods. Montana State University, David Sands
Mineral deficiency in the soil translates into mineral deficient foods produced in such soils. Increasing awareness of deficiencies using selected barley indicator mutant lines is one advance in this area. Included are indicators for deficient soils for zinc, iron, selenium, magnesium, and manganese. Montana State University, David Sands
Our research includes collaborations with one scientist from this W-3002 group and seven external scientists. Nineteen postdoctoral fellows, graduate students, undergraduate students, and staff were advised and supervised. University of Nebraska, Lincoln, Janos Zempleni
Nineteen papers, three reviews, and six meeting abstracts were published and 13 presentations were delivered. A public website was created that disseminates information about dietary microRNAs (see below, Chiang et al.). External grant support totaling $2.2 million direct costs per year were secured. University of Nebraska, Lincoln, Janos Zempleni
Metabolic syndrome (MetS) substantially increases the risk for developing nonalcoholic steatohepatitis, a liver disorder estimated to affect 70 million Americans. No validated therapies for NAFLD exist although long-term supplementation with ?-tocopherol (?-T) has been shown to reverse histological and biochemical evidence of NAFLD in humans. Unfortunately, supplementation with ?-T reduces ?-T status and its reactive nitrogen species scavenging ability. Ohio State University, Richard Bruno
Our pharmacokinetic studies examining ?-T bioavailability in MetS adults aim to reduce NASH risk by defining dietary ?-T requirements without inducing adverse effects on ?-T status. Our work provides the first evidence that dietary ?-T requirements for MetS adults are higher than that of healthy individuals, and are attributed to impairments in ?-T trafficking at the level of the small intestine and/or the liver. Ohio State University, Richard Bruno
Thus, not only are outreach efforts needed to ensure that healthy individuals meet the RDA for ?-T, which is currently achieved by <10% of Americans, additional efforts are needed to foster higher intakes among those with MetS. Ohio State University, Richard Bruno
Our research group seeks to identify nutrient deficiencies in the populations where we work and to contribute to education and policies for improving these deficiencies. Much of the data we collect about marginal intakes internationally can be applied to understanding marginal intakes in the developed world. Oklahoma State University, Barbara Stoecker
We continue to contribute to the international data on associations between dietary iodine intakes of pregnant women and concentrations of iodine in breast milk. Oklahoma State University, Barbara Stoecker
Effects of skin color on vitamin D status are being quantified. We also are monitoring infant and young child feeding practices and investigating adoption of agricultural practices to enhance growth and development of infants in the first two years of life. Our work supports the important links between agriculture, nutrition and health critical for continued development in Sub-Saharan Africa. Oklahoma State University, Barbara Stoecker
Diabetes is a major health issue and is one of the leading causes of death in the US and worldwide. Providing pre-diabetic individuals with dietary approaches that can help maintain their blood glucose is very important in delaying the progression of this chronic condition. Oklahoma State University, Edralin Lucas
Mangoes contain relatively high amounts of fiber, antioxidants ?-carotene and vitamin C, and phenolic compounds which have been well documented to play a role in the prevention of many chronic diseases. Our animal study demonstrated that mango pulp supplementation modulated the gut microbiota which may be important in preventing some of the adverse effects of high fat diets and improving gut tolerance. Oklahoma State University, Edralin Lucas
Diet plays an important role in mitigating the development and progression of several cancers, including prostate and breast. This research demonstrates that nutritional strategies that decrease oxidative stress, inflammation, DNA damage and/or target aberrant epigenetic alterations, such as acetylation and methylation, in prostate and breast cancer have the potential to dramatically reduce the incidence of prostate cancer. Oregon State University, Emily Ho
Our research has demonstrated that traditional food patterns from diverse cultures, including Northwest Indian tribes, have diverse nitrate concentrations (range ~350 to 100 mg/day nitrate) representing a rank order of nitrate concentrations of Mediterranean>Chinese>Japanese >American = Indian dietary patterns. Oregon State University, Norman Hord
The upper range of these nitrate intakes has, in clinical studies, been demonstrated to produce cardioprotective benefits. These ecological data are consistent with a potential contributory role of dietary nitrate intake to cardiovascular risk in these cultures. These data suggest that consumption of high nitrate plant foods may offer an inexpensive addition to cardioprotective dietary patterns. Oregon State University, Norman Hord
Dietary bioactive ingredients like prebiotics are beneficial to bone and help protect against shortfall minerals in the diet, especially calcium. Purdue University, Connie Weaver

Date of Annual Report: 07/19/2016

Report Information

Annual Meeting Dates: 05/24/2016 - 05/25/2016
Period the Report Covers: 10/01/2015 - 09/30/2016

Participants

Andrade, Juan (Chair) (jandrade@illinois.edu) – University of Illinois, Urbana-Champaign
Bruno, Richard (Bruno.27@osu.edu) – Ohio State University
Ho, Emily (emily.ho@oregonstate.edu) – Oregon State University
Lindshield, Brian (Secretary) (blindsh@ksu.edu) - Kansas State University
Liu, Yanhong (yahliu@ucdavis.edu) - University of California, Davis
Park, Yeonhwa (ypark@foodsci.umass.edu) – University of Massachusetts
Sands, David (uplds@montana.edu) – Montana State University
Teske, Jennifer (teskeja@email.arizona.edu) - University of Arizona
Vanamala, Jairam (juv4@psu.edu) - Pennsylvania State University
Zempleni, Janos (jzempleni2@unl.edu) - University of Nebraska, Lincoln
Shipka, Milan (Administrative Advisor) (mpshipka@alaska.edu) – University of Alaska, Fairbanks (via Skype)
Chester, Deirdra (USDA NIFA National Program Leader) (dnchester@nifa.usda.gov) (via Skype)

Brief Summary of Minutes

Brief Summary of Minutes of Annual Meeting

Meeting was called to order on May 24 and May 25, 2016 at 8:00 a.m. CST

Welcome and Introductions (May 24) – Participants were welcomed by the host and Meeting Chair, Dr. Juan Andrade, Food Science and Human Nutrition, University of Illinois, Urbana-Champaign; Dr. Sharon Nickols-Richardson, Department Head, Food Science and Human Nutrition, University of Illinois, Urbana-Champaign; and Dr. Peter Schiffer, Vice Chancellor for Research, University of Illinois, Urbana-Champaign.

Executive session (May 25) – Dr. Brian Lindshield was elected by the members present to be the chair of the next annual meeting. He will coordinate next year’s W3002 meeting in either Corvallis or Portland, Oregon; targeted for some time in summer 2017. Jennifer Teske, University of Arizona, was elected as secretary.

Dr. Andrade described the meeting objectives, timeline, and responsibilities.

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:

Emily Ho, Oregon State University – “Nutrient/environmental interactions for health”

David Sands, Montana State University – “Nutricrops: High lysine tomatoes and potatoes, and low glycemic crops”

Yeonhwa Park, University of Masachusetts – “Aging and obesity research using Caenorhabditis elegans”

Janos Zempleni, University of Nebraska, Lincoln – “The delivery of functional RNA species by dietary exosomes”

Jairam Vanamala, Pennsylvania State University – “Purple, not white, potato can prevent and reverse chronic colonic inflammation in the human-relevant pig model”

Brian Lindshield, Kansas State University – “Assessing the nutritional quality of fortified-blended foods: Field trial design, screening and baseline results”

Richard Bruno, Ohio State University – "Antiinflammatory therapeutic activities green tea for nonalcoholic steatohepatitis"

Jennifer Teske, University of Arizona – “Poor sleep and hedonic food intake”

Juan Andrade, University of Illinois, Urbana-Champaign – “Soy nanoparticles as delivery vehicles of vitamin D”

In addition, faculty members from the Food Science and Human Nutrition Department whose projects relate to the objectives of W3002 project presented and discussed their projects with members. These were:

Anna Arthur – “Dietary patterns and Cancer”

Hannah Holscher – “Impact of diet on the human gastrointestinal microbiome”

John W. Erdman Jr. – “Lycopene: Does this tomato bioactive play a role in prostate cancer risk”

Youngsoo Lee – “Microencapsulation of bioactive compounds – resveratrol & tributyrin”

Zeynep Madak-Erdogan – “Systems biology of estrogen receptor action in metabolism and breast cancer”

Hong Chen – “Soy consumption and Epigenetic Regulation of Gene Expression in preclinical models of colon cancer”

At the end of each presentation a short discussion took place. Also, in the second day of meetings the group had more time for discussion.

Our new administrative advisor, Dr. Milan Shipka, connected via Skype to introduce himself and provide guidance on the project’s renewal.

Included in the agenda were tours to the Sustainable Farm and Blue Waters.

Discussions among members – Several of the topics presented stirred discussion among the members including:

How can we address global nutrition problems through R&D and biotechnology? Dave Sands talked about the breeder’s dilemma in addressing this question.

Yeonhwa Park led the discussion about the virtues and limitations of using C. Elegans in nutrition research.

Emily Ho led the discussion about the need for models of Zinc metabolism. She argued the use of the zebrafish model to expand nutrition research with metals.

Time was also spent discussing nutrition drivers such as prevention of chronic disease. For example, several members of the project and invited guests discussed the role of nutrients and bioactives in the etiology of chronic disease such as cancer, diabetes, cardiovascular disease.

The studies of some members also focused on global problems such as food aid and vitamin D deficiency. These topics were discussed in the light of low resource settings and availability of tools to assess nutrition status.

The group discussed research priorities and membership for the upcoming renewal and agreed that the next annual meeting would include significant discussion, assembly of writing teams and preparation of renewal.

The meeting adjourned on May 25, 2016 at, 2:00 p.m.

Accomplishments

Efforts from the members focused on new models to assess nutrition outcomes, bioavailability of vitamin D from specialized soy-based nanoparticles, effects of bioactives/nutrients from whole foods or extracts on oxidative stress and inflammation pathways, the role of new bioactives such as microRNA species, and how conditions such as sleep deprivation can affect food consumption. These are in alignment with objective 1 and 2 below. Members of the W3002 Multistate project have been vastly productive during the past reporting period as evidenced by the dissemination of new products, technologies, and knowledge through peer-review publications, white papers, book reviews, presentations at symposia, conferences, and in the media. Project objectives are listed below along with scholarship activities from each of the represented institutions. <ol> <li>Determine the bioavailability (absorption, distribution, metabolism, elimination) of nutrients and other food components.</li> <li>Evaluate the bioactivity of nutrients and other food components in order to elucidate their underlying protective mechanisms.</li> </ol> University of Arizona (Jennifer Teske). The laboratory focuses on the negative impact of poor sleep and low physical activity to health. We investigate the relationship between sleep curtailment due to environmental noise and metabolism in a rodent model. We expanded these studies to address whether extending sleep with pharmacological aids would improve metabolic markers. These studies revealed the improvements were dependent upon the mechanism of action of the pharmacological agents. We also addressed the role of hedonic food intake to weight gain associated with poor sleep since hedonic intake contributes to overall obesity. We validated that consumption of highly palatable human snack items caused obesity in mice. In an established rodent model of binge eating, we observed that preference for palatable food moderated the effect of sleep deprivation on the hedonic intake. Finally, we investigated whether poor sleep during the postsurgical period worsened recovery and motor function since high noise levels in hospitals disrupts sleep. We validated that sleep deprivation worsened postsurgical recovery in rats. Sleep deprived rats have more surgical complications and pain and lower skeletal muscle strength and physical activity that persisted despite resumption to normal sleep patterns. OUTPUTS: i) Determined central orexin-A injections promote gamma power. ii) One oral presentation on research presented by the doctoral student at an annual scientific conference. iii) two poster presentations on research presented by a postdoctoral fellow and a high school student. iv) Research opportunities provided to 1 postdoctoral fellow, 1 doctoral student, 2 International students, 3 Master’s degree students, 3 undergraduates, and 1 high school student.  v) Outreach presentation to Zonta International. vi) Hosted 2 international students to further collaborations.  MILESTONES: To test whether exposure to environmental noise disrupts sleep in female rats in a manner similar male rats and whether the effect of sleep deprivation on hedonic food intake differs between male and female rats. OUTCOMES: i) Surgery training and proficiency provided for 2 international students and 1 doctoral student. ii) Presentation skills training for oral and posters for 1 doctoral student, 5 Master’s degree students, 3 undergraduates, and 1 high school student. University of California, Berkeley (Barry Shane). SUMMARY OF PROGRESS: 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 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. Methylmalonic acid (MMA) is a by-product of propionic acid metabolism through the vitamin B12 (cobalamin)-dependent enzyme methylmalonyl CoA mutase. Elevated MMA concentrations are a hallmark of several inborn errors of metabolism and indicators of cobalamin deficiency in older persons. In a genome-wide analysis of 2,210 healthy young Irish adults (median age 22 years), we identified a strong association of plasma MMA with SNPs in 3-hydroxyisobutyryl-CoA hydrolase (HIBCH) and acyl-CoA synthetase family member 3 (ACSF3). These loci accounted for 12% of the variance in MMA concentration. The most strongly associated SNP (HIBCH rs291466) causes a missense change of the initiator methionine codon to threonine. Surprisingly, the resulting variant is associated with increased expression of HIBCH mRNA and encoded protein. These homozygotes had, on average, 46% higher MMA concentrations than methionine-encoding homozygotes in young adults with generally low MMA concentrations.   The association between MMA levels and HIBCH rs291466 was also highly significant in a replication cohort of 1,481 older individuals (median age 79 years) with elevated plasma MMA concentrations (0.34 [0.24–0.51] mmol/L. Additionally, in a longitudinal study of 185 pregnant women and their newborns, the association of this SNP remained significant across the gestational trimesters and in newborns. University of Connecticut (Ji-Young Lee; Sung Koo). SUMMARY OF PROGRESS: We determined if astaxanthin (ASTX), a xanthophyll carotenoid, can prevent obesity-associated metabolic abnormalities, inflammation and fibrosis in diet-induced obesity (DIO) and nonalcoholic steatohepatitis (NASH) mouse models. Male C57BL/6J mice were fed a low-fat (6% fat, w/w), a high fat/high sucrose control (HF/HS; 35% fat, 35% sucrose, w/w), or a HF/HS containing ASTX (AHF/HS; 0.03% ASTX, w/w) for 30 wk. To induce NASH, another set of mice was fed a HF/HS diet containing 2% cholesterol (HF/HS/HC) or HF/HS/HC with 0.015% ASTX (AHF/HS/HC) for 18 wk. Compared to LF, HF/HS significantly increased plasma total cholesterol, triglyceride and glucose, which were lowered by ASTX. ASTX decreased hepatic mRNA levels of markers of macrophages and fibrosis in both models. The effect of ASTX was more prominent in NASH than DIO mice. In epididymal fat, ASTX also decreased macrophage infiltration and M1 macrophage marker expression and inhibited hypoxia-inducible factor 1-alpha and its downstream fibrogenic genes in both mouse models. ASTX significantly decreased tumor necrosis factor alpha mRNA in the splenocytes from DIO mice upon lipopolysaccharides stimulation, compared with those from control mice fed a HF/HS diet. Additionally, ASTX significantly elevated the levels of genes that regulate fatty acid b-oxidation and mitochondrial biogenesis in the skeletal muscle, compared with control obese mice, whereas no differences were noted in adipose lipogenic genes. Our results indicate that ASTX inhibits inflammation and fibrosis in the liver and adipose tissue and enhances the skeletal muscle's capacity for mitochondrial fatty acid oxidation in obese mice. Evidence indicates that berry anthocyanins are anti-atherogenic, antioxidant, and anti-inflammatory. However, berries differ vastly in their anthocyanin composition and thus potentially in their biological and metabolic effects. The present study compared the hypolipidemic, antioxidant, and anti-inflammatory properties of blueberry (BB), blackberry (BK), and blackcurrant (BC) in a diet-induced obesity (DIO) mouse model. Male C57BL/6J mice were fed a high fat (HF; 35 % fat by weight) as control or a HF diet supplemented with 5 % freeze-dried BB, 6.3 % BK or 5.7 % BC for 12 weeks. The different amounts of berries were added to the basal control diet to achieve the same total anthocyanin level. Plasma lipids, antioxidant status, and pro-inflammatory cytokines were measured. The expression of genes involved in antioxidant defense, inflammation, and lipid metabolism were determined in the liver, epididymal adipose, proximal intestine, and skeletal muscle. Histological analysis was performed to identify crown-like structure (CLS) in epididymal fat pads to determine macrophage infiltration. No differences were noted between the control and berry-fed groups in plasma liver enzymes, insulin, glucose, ferric reducing antioxidant power, superoxide dismutase and tumor necrosis factor a. However, BK significantly lowered plasma triglyceride compared with the HF control and other berries, whereas BC significantly reduced F4/80 mRNA and the number of CLS in the epididymal fat pad, suggestive of less macrophage infiltration. The present findings provide evidence that the berries varying in anthocyanin composition differentially affect plasma lipids and adipose macrophage infiltration in DIO mice, but with no differences in their antioxidant capacity and anti-inflammatory potential. University of Illinois, Urbana-Champaign (Juan Andrade). SUMMARY OF PROGRESS: Our research focuses on developing technologies or approaches to improve nutrition in vulnerable populations in the US and abroad. Recently, we have been working on the characterization of legume-based protein nanoaggregates to improve stability and delivery of fat soluble bioactives and nutrients. OUTPUTS: Our team found that a legume-based nanoparticle system could be advantageous for the delivery of fat-soluble compounds that can address nutrition gaps due to its enhanced stability and bioavailability. Nanoaggregates containing vitamin D were characterized in terms of stability and bioavailability in animal models. King Saud University in Saudi Arabia is a major partner in this research endeavor. OUTCOMES (Change in knowledge) The novel legume-based protein nanoaggregates can disperse vitamin D and enhance its bioavailability and bioefficacy in vivo. Vitamin D dispersed in legume-based protein nanoemulsion was more stable against UV challenge than vitamin D dispersed water or in water containing lecithin. Reduced particle size enhanced vitamin D bioefficacy. Kansas State University (Brian Lindshield). SUMMARY OF PROGRESS: We are completing a 20-week field trial assessing micronutrient and anthropometric outcomes from consuming new extruded micronutrient fortified-blended foods (FBFs), which are commonly used for food aid, in Tanzanian children under 5 years of age. Two white and one red blend of sorghum-cowpea with, white sorghum-soy, extruded corn-soy blend, will be compared to CSB+ (current USAID FBF). In addition, a group will not receive FBFs until after the study to serve as a control. OUTPUTS: We have produced useful data and are preparing a protocol manuscript to disseminate what we have learned from observations and experiences in setting up and preparing for the field trial. MILESTONES: We have completed midline data collection, with end line data collection set to start on 6/14. OUTCOMES: Anecdotal reports from mothers, and some preliminary evidence suggests that health outcomes are improving in children that are receiving FBFs. University of Maine (Dorothy Klimis-Zacas). SUMMARY OF PROGRESS: One project targeted Perivascular Adipose Tissue (PVAT) that has been shown to play important roles in regulating vascular tone and linking local and systemic vascular inflammation and is especially expanded during Obesity. We examined the impact of PVAT on Phenylephrine-mediated vasoconstriction in the aorta of Obese Zucker rats (OZR) and their lean littermates (LZR), (models of the Metabolic Syndrome) by comparing aortic rings with or without PVAT. Subsequently, we placed OZR and LZR on a control (C) or an 8% wild blueberry (WB) diet and evaluated the effect of WB consumption on such response.  PVAT-released adipokine concentrations were also measured as a function of WB diet.  Maximal constrictor force (Fmax) in aortic rings without PVAT was significantly lower in OZR-C compared to LZR-C (0.41 ± 0.05 and 0.71 ± 0.06 g, respectively). Following WB diet, Fmax significantly increased in OZR (0.54 ± 0.06 g). In aortas with intact PVAT, Fmax was significantly lower in all groups (0.31 ± 0.06 OZR-C, 0.30 ± 0.05 OZR-WB, 0.29 ± 0.03 LZR-C and 0.30 ± 0.04 g LZR-WB), but no difference was observed between treatments. PVAT concentrations of MCP-1, TNF-alpha and adiponectin, were significantly higher in OZR compared to LZR (+102%, +108%, and +45%, respectively). Following WB diet, PVAT concentrations of IL-8 were significantly lower in both OZR (–37%) and LZR (–30%), while adiponectin concentrations significantly increased in both OZR (+11%) and LZR (+16%).  MCP-1 concentrations significantly decreased (–31%) in the PVAT of OZR on WB diet. OUTCOME project 1) Wild blueberry consumption appears to attenuate local inflammation in PVAT, which may impact systemic vascular inflammation and endothelial function in the Metabolic Syndrome.  Another project targeted Cardiovascular Atherosclerotic Disease (CAD) which is a major public health problem and is responsible for most deaths in developed societies and developing countries worldwide. Atherosclerosis is a chronic inflammatory, progressive disease of the large arteries that can lead to CVD and stroke. Angiogenesis is the formation of new capillary blood vessels from existing ones and endothelial cell migration and proliferation contribute to the development of angiogenesis; critical in the early stages of atherosclerosis.  Wild blueberries (Vaccinium angustifolium) are rich in anthocyanins (ACNs) and phenolic acids (Phen) having an exceptional ranking for antioxidant capacity compared to other berries and fruits. Past research (animal models) in our laboratory has documented the beneficial effects of wild blueberries to attenuate inflammation and improve endothelial function in the Metabolic Syndrome. During the last year, Dr. Klimis-Zacas’s laboratory team have studied human umbilical vein endothelial cells (HUV-EC-C [HUVEC] (ATCC® CRL-1730™)) and the effect of ACN and Phen fractions and their combination (extracted from wild blueberry powder) to determine whether and how they operate to alter cell migration and angiogenesis. Preliminary results on the effect of ACN and Phen fractions on cell migration show a differential response to endothelial cell migration based on the type of fractions and fraction concentration. Studies on angiogenesis also document a concentration-depended effect, critical concentrations at which angiogenesis is modulated and differential response based on the different fractions (ACNs or Phen). Future experiments will explore the mechanism of ACN and Phen fraction signaling to alter endothelial migration and angiogenesis related to atherosclerosis, critical for public health.   OUTPUTS: Results from the period covered in this report were disseminated at the Experimental Biology meetings in Boston in April 2015 and will be disseminated at the American College of Nutrition in San Diego, November 2016 and at the Experimental Biology Meetings, Boston, April 2017, and through published articles, presentations at invited lectures and through the media (newspaper articles, blogs and interviews). University of Massachusetts (Yeonhwa Park). We have studied various bioactive food components to improve bioaccessibility and bioavailability as well as determine their role in obesity, aging, inflammation, and colon health. We have continued our efforts to improve bioaccessibility and bioavailability for lipophilic bioactives (such as fucoxanthin, luteolin, tangeretin, nobiletin, conjugated linoleic acid, and fat-soluble vitamins) by developing and evaluating various delivery systems. Moreover, to test bioactives on aging and obesity, we established Caenorhabditis elegans models and tested cranberry phenolics on obesity and piceatannol on ageing. University of Missouri (Ingolf Gruen). ACCOMPLISHMENTS: Neem (Azadirachta indica) is an evergreen tree cultivated in various parts of the Indian sub-continent. It has been in use for centuries in the Indian folk medicine for its therapeutic value. Given the prominent role it has played in curing diseases of the villagers over centuries, it has been hailed as a “divine” tree, a “village dispensary” and “nature’s drugstore”. Today, extensive research has shown that it may have anti-cancer, anti- diabetic, anti-inflammatory, anti-ulcerogenic, and anti-microbial effects. However, there are significant gaps in the scientific literature about the compounds contributing to its medicinal potential. In this study, select flavonoids, namely flavonols: myricetin, quercetin, and kaempferol are quantified by an HPLC method in Neem powder and leaves as well as green and black tea leaves for comparison. It was observed that Neem dry powder had significantly higher (p< 0.05) flavonol content at 11.146 ±1.09 mg/g than green and black tea 5.782 ± 0.723 mg/g and 5.599 ± 0.484 mg/g respectively. The tea infusions of the plant materials also yielded similar results. The total phenolics were found to be the highest in green tea infusion because of its high flavan-3-ol content, followed by black tea, Neem powder, and Neem tea cut leaves with values of 174.55 ± 12.85, 106.85 ± 6.511, 55.60 ± 7.90 and 87. 35 ± 7.42 mg/g respectively. DPPH and FRAP antioxidant assays yielded contradictory results which can be explained by the very principle of these assays. It was also observed that the particle size of ground tea leaves significantly influenced the yield of flavonols, phenolics, and antioxidant assay. Montana State University (David C. Sands). SUMMARY OF PROGRESS: The problem of developing food crops with higher nutritional value is that the selection of nutritional value often conflicts with selection for traits necessary for high agronomic yield. This is a problem with our field crops - peas, potatoes, wheat, oat and barley. We have developed rapid selection parameters for potatoes that are lower in glycemic index than the main commodity varieties. We now have 7 cultivars being tested for agronomic production and of these, we will choose several for human glycemic index testing. One low glycemic cultivar has been selected for its higher lysine level, and this is being crossed with other cultivars. Similarly, a series of high lysine tomato cultivars have been similarly selected, and these are in the F2 cross stage. We have developed a high-throughput screening platform for the oilseed crop, Camelina sativa yielding four lines that are higher, much higher in the omega-3;omega-6 fatty acid ratio than canola, corn, soy, or safflower oils. Proatina, a naked oat, is being developed for use as a high protein/low glycemic substitute for rice because of its nutritional value compared to rice (2.5x higher in protein, much lower glycemic index, absence of arsenates), and by virtue of its low pesticide demand and dryland habit it is a more ecologically sustainable to produce. The market target is the global community of Type II diabetics, especially those that use rice as a food staple. Agriculture, especially smallholder farmers, and gardeners, needs an inexpensive soil deficiency assay. This is clearly a problem not only in Montana but almost everywhere. We saw a need for a simple on-farm assay in our Gates-funded project in western Kenya where we successfully developed a dirt cheap biological control of Striga for 500 women smallholder farmers.  Our seed sentinel (a soil bioavailability assay of trace elements and macro elements) using mutant barley lines deficient in uptake of 9 specific elements have been realized in greenhouse studies and field tests are being developed. The goal is to have an assay of all 15 elements arranged as mutant barley seeds on a string to be planted and read in 3 weeks. These are being prepared as seed strips to be tested this year in a number of locations.  Our research has continued on plasmid curing, a potentially important approach to reducing plasmid associated antibiotic resistance that is a mounting concern for the medical industry. Preliminary experiments on lambs with scours have shown significant results. University of Nebraska, Lincoln (Janos Zempleni). SUMMARY OF PROGRESS:  microRNAs are small sequences of RNA (about 22 nucleotides). They repress genes through binding to complementary sequences in mRNA, thereby causing mRNA degradation of halting translation. A large fraction of microRNAs in body fluids and milk is encapsulated in exosomes (nanoparticles), thereby conferring protection against degradation and providing a mechanism for cellular uptake by endocytosis. Note that traditionally microRNAs have been considered endogenous regulators of genes, i.e., microRNAs synthesized by an organism regulate the genes in that organism. We have refuted this paradigm through the following discoveries. 1) Human intestinal cells transport cow’s milk exosomes by endocytosis and secrete milk exosomes and/or microRNA cargos across the basolateral membrane. 2) Glycoproteins on the surface of exosomes and cells are essential for endocytosis. 3) Milk (and chicken egg) exosomes are delivered to peripheral tissues and circulating cells, facilitated by endocytosis of exosomes in human vascular endothelial cells. 4) Postprandial concentrations of milk and egg microRNAs are biologically effective in humans and elicit changes in gene expression in circulating lymphocytes. 5) Endogenous synthesis of microRNAs is insufficient to compensate for dietary microRNA depletion. 6) Depletion of dietary microRNAs elicits phenotypes such as impaired fecundity, aberrant purine metabolism, and possibly altered immune function and changes in the gut microbiome. OUTCOMES: Twelve postdoctoral fellows, graduate students, undergraduate students, and staff were advised and supervised. Nine papers, one review, seven meeting abstracts and four outreach publications (newspaper, magazines) were published, and three presentations were delivered. A public website was created that disseminates information about dietary microRNAs (see below, Chiang et al.). Our research includes collaborations with one scientist from this W-3002 group and six external scientists. The Ohio State University (Richard Bruno). ACCOMPLISHMENTS: Green tea catechins protect against inflammation during nonalcoholic steatohepatitis, but the mechanisms by which they inhibit NFkB activation are unknown. We, therefore, tested the hypothesis that green tea extract (GTE) mitigates NFkB activation in an obese rodent model of NASH in a Nrf2-dependent manner. We conducted a dietary intervention study in wild-type (WT) and Nrf2-knockout mice in which they received a high-fat diet that was devoid of, or supplemented with, GTE for 8 wk. Compared with WT mice, Nrf2-KO mice had exacerbated NASH pathology that was evidenced by increased liver steatosis, hepatocyte ballooning, biomarkers of liver injury, and NFkB activation and expression of pro-inflammatory cytokines (P<0.05).  In WT mice, GTE increased Nrf2 and Nqo1 mRNA expression while also lowering histological and biochemical parameters of liver injury and NFκB-dependent inflammatory responses.  Similarly, GTE treatment in Nrf2-KO lowered histological parameters of NASH along with NFκB activation and hepatic lipid peroxidation.  GTE also increased hepatic mRNA expression of TLR4 and TNFR1 in both WT and Nrf2-KO mice. Collectively, these findings support that the anti-inflammatory mechanism of GTE is mediated through a mechanism independent of intracellular Nrf2 signaling and that its inhibition on NFkB activation may be  mediated through extracellular signaling downstream of TLR4 and/or TNFR1. Oklahoma State University (Edralin Lucas). SUMMARY OF PROGRESS: Consumption of high-fat (HF) diet is one of the major causes of obesity, a major global health problem. Obesity has put many individuals at risk for co-morbidities including type 2 diabetes (T2D), heart disease, hypertension, as well as gastrointestinal disorders.  One of the underlying links between HF diet and obesity are changes in gut microbiota. For this reporting period, we investigated dietary approaches (i.e. freeze-dried mango pulp and wheat germ supplementation) on the gut microbiota and their fermentation products, and its impact on body composition, glucose homeostasis, and gut inflammatory markers. Mango supplementation did not reduce body weight or fasting blood glucose, but a dose-dependent increase in microbial fermentation was observed with mango supplementation. Furthermore, mango supplementation modulated gut inflammation, as observed with an increase in colonic interleukin (IL)-10 gene expression. Our preliminary data also indicate that wheat germ has some positive effects on the gut microbial population. We are also currently examining the effects of freeze-dried mango supplementation on postprandial responses in healthy men. Results during this reporting period were presented to four national meetings, two local professional meetings, and one public meeting. Oklahoma State University (Barbara Stoecker). SUMMARY OF PROGRESS: Our laboratory has continued studies on micronutrient deficiencies and human health.  Analyses of bioactivity of zinc, iodine, and vitamin D in three studies in Ethiopia were the focus of work within the last year.  From a small randomized-control trial of 20 mg zinc (as zinc sulfate) for 17 days, antioxidant activity and oxidative stress biomarkers in blood and urine were assessed.  Ferric reducing the ability of plasma (FRAP), 8-hydroxydeoxyguanosine (8-OHdG), malondialdehyde (MDA), super oxide dismutase (SOD), urinary 8-isoprostane and other biomarkers measured were not changed significantly in response to zinc supplementation.  However, trends in these antioxidant activity biomarkers and oxidative stress markers merit further analyses.  Increases representing oxidative damage were observed in 8-OHdG and 8-isoprostane in the placebo groups. Metabolomics analyses conducted by multistate collaborators revealed significant differences in plasma metabolite profiles before and after zinc supplementation.  Identification and validation of these endogenous metabolites continues.  A community-based, randomized supplementation trial was conducted which compared effects of a potassium iodide capsule (225 ug iodine) daily for 26 weeks versus providing appropriately iodized salt for 26 weeks.  Women were enrolled in the trial within one week of delivery.  Maternal thyroid stimulating hormone (TSH) and goiter rate were significantly reduced following capsule ingestion compared to iodized salt, but infant thyroxine, TSH and visual information processing were not impacted by the method of iodine delivery. Median breast milk iodine concentration was adequate to provide the calculated dietary requirement for iodine, and median urinary iodine of infants also was adequate with both methods of delivery.   Plasma 25(OH)D concentration and its relations with dress habits, sun-seeking behavior and skin reflectance were assessed among 120 Ethiopian lactating women.  Skin reflectance was measured using a portable colorimeter.  At baseline, mean (SD) plasma 25(OH)D was 48.5 (14.6) nmol/L.  Values ranged from 23 to 102 nmol/L.  In 55% of women, 25(OH)D was between 30 and Oregon State University (Emily Ho). SUMMARY OF PROGRESS: Zinc and chronic disease:  Recently we have found that zinc status is compromised with age.  Zinc deficiency causes immune cells to be sensitized to inflammation and also occurs in older animals and humans.  We have also examined the impact of zinc deficiency on developmental outcomes, gut microbiota oxidative stress and inflammation induced by arsenic.   For these studies, we have developed a zebrafish and rodent models to examine the interaction among zinc and arsenic.   Plant-derived phytochemicals and cancer: We have found that sulforaphane, a chemical found in cruciferous vegetables inhibits histone deacetylases, decreases DNA methyltransferase 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. Supplemental SFN did show benefits in decreasing cell proliferation markers and histone deacetylase 3 expression in breast tissue in women previously diagnosed with breast cancer.  Studies in prostate cancer patients are ongoing. OUTPUTSs:  i) Identify new risk factors in prostate cancer and offer novel dietary modifications to reduce the incidence of prostate cancer; ii) Establish low cruciferous vegetable intake as a risk factor for the development of prostate cancer by altering histone modifications and cell proliferation pathways; iii) Gain knowledge of the mechanisms behind the health benefits of micronutrients and phytochemicals such as zinc and compounds derived from cruciferous vegetables; iv) Establish low dietary zinc as risk factor for inflammatory processes, DNA damage and cancer risk and identify new biomarkers for human zinc deficiency; v) Establish function of zinc and changes in zinc metabolism with age. Oregon State University (Norman Hord). SUMMARY OF PROGRESS: This report summarizes progress to address potential health benefits of dietary nitrate and nitrite that are primarily derived from plant foods in the human diet. Since nitrate and nitrite have been associated with risk of methemoglobinemia in newborns and gastrointestinal cancer risk in adults when consumed in processed meats, nitrate concentrations in drinking water are regulated by the U.S. EPA. With regard to human dietary intake, vegetable and fruit intake constitute the source of 80% of nitrate in the diet, and these food groups are positively associated in epidemiological studies with decreased risk of cardiovascular disease and low bone mass. Notably, human dietary nitrate intakes have, in clinical studies, been demonstrated to produce cardioprotective benefits such as blood pressure lowering and platelet inhibition.  Our projects work across the spectrum from food-based analysis to zebrafish and rodent model systems to assess dietary exposures, potential toxicities, and potential health benefits, respectively. Manuscripts representing one project have been accepted for publication, one book chapter is being published, one manuscript is under revision for publication, and two are in preparation. Project 1: The objective of this project was to test the hypothesis that nitrate or nitrite exposure causes increases in mortality and altered development in zebrafish (Dano rerio) We exposed zebrafish embryos or larvae to 10 fold increasing concentrations of sodium nitrate from 0.01-100 mM, sodium nitrite ranging from 0.001-10 mM or sodium chloride as a control at each concentration. Fish were exposed for 6 hours post fertilization (hpf) and then evaluated for mortality and 22 additional developmental malformations or endpoints at 24 hours and 5-day endpoints. OUTCOMES: Project 1) Our data indicate that only nitrate and nitrite concentrations of 10 mM and 10 µM and were associated with toxicity, respectively.  Concentrations of nitrite observed to be toxic were also associated with abnormal photomotor responses while nitrate elicited hyperactive photomotor responses at the larval stage at 0.01, 1 and 10 µM. These data indicate minimal risk of mortality and developmental toxicity due to nitrate and nitrite concentrations except at very high concentrations in the environment.  Project 2) Evidence suggests a diet rich in fruits and vegetables reduce bone loss in older adults, although the specific compounds responsible are not known. The objectives of project 2 were to investigate the ability of dietary nitrate, a component in leafy green vegetables, to reduce bone turnover and loss as well as alter the gut microbiome structure and composition in an ovariectomized (OVX) rat model. Outcome 2: Using OVX rats fed either control diet with no added nitrate, control diet plus 0.1 mmol nitrate/kg bw/day, or 1.0 mmol nitrate/kg bw/day corresponding to OVX control, low- (LDN) and high- (HDN) dose nitrate for three weeks, we found that OVX resulted in increased bone turnover, cancellous bone loss, and changes in gut microbiome composition and structure.  However, three weeks of LDN or HDN supplementation had no beneficial effect on the skeleton or gut microbiome of an ovariectomized rat model of bone loss. These in vivo data indicate dietary nitrate does not slow bone turnover and loss in an OVX rat model of osteoporosis.  Project 3) There is a dearth of widely available information on dietary nitrate and nitrite sources for use by epidemiologists and clinicians.  Since the USDA National Nutrient Database for Standard Reference does not contain nitrate or nitrite, we established a relationship with scientists at the USDA Agricultural Research Service's Nutrient Data Laboratory to develop a new, special interest database for food nitrate and nitrite concentrations (Pamela R. Pehrsson, PhD, Research Leader, Juhi R. Williams, Nutritionist; Proposal for Development of USDA Special Interest Database on Nitrites and Nitrates in US Foods Nutrient Data Laboratory, Beltsville Human Nutrition Research Center, ARS-USDA Beltsville, MD 20705).  Outcome 3) We would be the pioneer contributors to this database by supplying nitrate and nitrite concentrations for ~300 foods.  Other laboratories will be invited to participate in a systematic, reference-based network providing analyses of nitrate and nitrite in foods. Purdue University (Connie Weaver). SUMMARY OF PROGRESS: Prebiotics enhanced mineral absorption in animal models and humans and bone parameters in animal models.  We completed an analysis of our study on the effect of soluble corn fiber (SCF) on bone calcium retention in postmenopausal women.  We used our novel approach of urinary excretion of the rare isotope, 41Ca, from pre-labeled bone to test the dose responsive effect of feeding 0, 10 and 20 g SCF/d.  SCF increased (p<0.01) bone calcium retention in a dose responsive manner.  We obtained funding to do metagenomics analysis for this study.

Publications

Publications Ahluwalia, A., Gladwin, M., Coleman, G.D., Hord, N.G., Howard, G., Kim-Shapiro, D., Lajous, M., Larsen, F., Lefer, D.J., McClure, L.A., Nolan, B.T., Pluta, R., Schechter, A., Wang, C.-Y., Ward, M.H., and Harman, J.L. (2016) Dietary nitrate and the epidemiology of cardiovascular disease: Report from a National Heart, Lung, and Blood Institute Workshop, Journal of the American Heart Association. 5(7). doi: 10.1161/JAHA.116.003402 Almajwal, AM, Abulmeaty, M, Andrade, JE. Efficacy of a novel food fortification system to combat vitamin D deficiency in rats. FASEB Journal (Abstract #LB267) Experimental Biology, Late breaking abstract, San Diego, CA, April 2016 Alu’datt, M, Rababah, T, Ereifej, K, Al-Hamad, M, Mhaidat, NM, Andrade, JE, Ayadi, WA. Probiotics in milk as functional food: characterization and nutraceutical properties of extracted phenolics and peptides from fermented skimmed milk inoculated with royal jelly. Journal of Food Safety. 35(4):509-22. (2015) Atwell LL, Beaver LM, Shannon J, Williams DE, Dashwood RH, Ho E. Epigenetic Regulation by Sulforaphane: Opportunities for Breast and Prostate Cancer Chemoprevention. Curr Pharmacol Rep. 2015 Apr 1;1(2):102-111 Atwell LL, Zhang Z, Mori M, Farris PE, Vetto JT, Naik AM, Oh KY, Thuillier P, Ho E, Shannon J. Sulforaphane Bioavailability and Chemopreventive Activity in Women Scheduled for Breast Biopsy. Cancer Prev Res (Phila). 2015 Dec;8(12):1184-9 Bailey RL, Looker AC, Lu Z, Fan R, Eicher-Miller HA*, Fakhouri TH, Gahche JJ, Weaver CM, Mills JM. B-vitamins and bone mineral density and risk of lumbar osteoporosis in older females in the U.S. Am J Clin Nutr   102:687-954, 2015. Bailey RL, Parker EA, Rhodes DG, Goldman JD, Clemens JC, Moshfegh AJ, Thuppal SV, Weaver CM. Estimating sodium and potassium intakes and their ratio in the American diet:  Data from the 2011-2012 NHANES.  J Nutr 2016 doi: 10.3945/jn.115.221184 Ballard KD, E Mah, Y Guo, RS Bruno, BA Taylor, JE Beam, DM Polk, PD Thompson (2016). Single low-density lipoprotein apheresis does not improve vascular endothelial function in chronically treated hypercholesterolemic patients. Int J Vascular Medicine, 2016: 4613202, 1-7. doi:10.1155/2016/4613202. Chiang K, Shu J, Zempleni J, Cui J. Dietary MicroRNA Database (DMD): an archive database and analytic tool for food-borne microRNAs. PLoS ONE 10(6):e0128089, 2015 Colmenares D, Q. Sun, P. Shen, Y. Yue, D. J. McClements, and Y. Park (2016) Delivery of Dietary Triglycerides to Caernohabditis elegans Using Lipid Nanoparticles: Nanoemulsion-Based Delivery Systems, Food Chem. 202: 451-457. Conley, MN, Maccha, A, Roberts, C, Sharpton, TJ, Urszula T. Iwaniec, UT, Hord NG. (2016) Increasing dietary nitrate has no effect on bone loss or gut microbiome in ovariectomized rats. Under revision for Molecular Nutrition and Food Research. Conley MN, Wong CP, Duyck KM, Hord N, Ho E, Sharpton TJ. Aging and serum MCP-1 are associated with gut microbiome composition in a murine model. (2016) PeerJ. Mar 31;4:e1854. doi: 10.7717/peerj.1854. eCollection 2016. Cordonier EL*, Adjam R*, Teixeira DC, Onur S, Zbasnik R, Read PE, Döring F, Schlegel VL, Zempleni J. (*contributed equally). Resveratrol compounds inhibit human holocarboxylase synthetase and cause a lean phenotype in Drosophila melanogaster. J Nutr Biochem, 2015: 26(11):1379-84. Doi: 10.1016/j.jnutbio.2015.07.004 Cordonier EL, Jarecke SK, Hollinger FE, Zempleni J. Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells. Eur J Pharmacol 2016:780:202-8. doi: 10.1016/j.ejphar.2016.03.052 Cui L, H. T. Cho, D. J. McClements, E. A. Decker, and Y. Park (2016) Effects of salts on oxidative stability of lipids in oil-in-water emulsion, Food Chem. 197: 1130-1135. Del Bo’ C, Porrini M, Campolo J, Parolini M, Lanti C, Klimis-Zacas D and Riso P, Effect of a single blueberry (V. corymbosum) portion on markers of antioxidant defense and oxidative stress in healthy volunteers following cigarette smoking: a randomized-controlled study, Food and Function, 2014, 5, 3107 Ferira AJ, Laing EM, Hausman DB, Hall DB, McCabe GP, Martin BR, Hill Gallant KM, Warden SJ, Weaver CM, Peacock M, Lewis RD. Vitamin D supplementation effects on insulin sensitivity and resistance in early pubertal white and black children.  J Clin Endocrinol Metab, 2016;101(4):1710-8. Doi: 10.1210/jc.2015-3687. Funaro A, Wu X, Song M, Zheng J, Guo S, Rakariyatham K, Rodriguez-Estrada MT, Xiao H (2016) Enhanced anti-inflammatory activities by the combination of luteolin and tangeretin, J. Food Sci. 81: H1320-1327. Gac L, Butterick TA, Duffy CM, Teske JA and Perez-Leighton CE. Role of the non-opioid dynorphin peptide des-Tyr-dynorphin (DYN-A2-17) in food intake and physical activity, and its interaction with orexin-A. 2016. Peptides. 76:14-18. Epub 2015 2015 Dec. 1. Gac L, Kanaly VR, Teske JA, Pinto MP and Perez-Leighton CE. Behavioral characterization of a model of differential susceptibility to obesity induced by standard and personalized cafeteria diet feeding. 2015. Physiology & Behavior. 152 (Pt A):315-322. Epub 2015 Oct. 9. Gashu D, Stoecker BJ, Adish A, Haki GD, Bougma K, Marquis GS. (2016) Association of serum selenium with thyroxin in severely iodine-deficient young children from the Amhara region of Ethiopia. Eur J Clin Nutr DOI: 10.1038/ejcn.2016.27. Gashu D, Stoecker BJ, Adish A, Haki GD., Bougma K, Marquis, GS. (2016) Ethiopian pre-school children consuming a predominantly unrefined plant-based diet have low prevalence of iron-deficiency anaemia.  Public Health Nutr 19:1834-1841.  DOI:10.1017/S1368980015003626.   Gashu D, Stoecker BJ, Bougma K, Adish A, Haki GD, Marquis GS. (2016) Stunting, selenium deficiency and anemia are associated with poor cognitive performance in preschool children from rural Ethiopia. Nutr J 15:1-8. DOI: 10.1186/s12937-016-0155-z. Hill Gallant KM, Weaver CM, Towler DA, Thuppal SV, Bailey RL. Nutrition in cardioskeletal health.  Adv Nutr, 2016;7(3):544-55. doi: 10.3945/an.115.011189. Hord, NG and Conley M. (2016) Regulation of dietary nitrate and nitrite: balancing essential physiological roles with potential health risks, In: Nitrates and Nitrites in Human Health and Disease, Editors: Joseph Loscalzo, M.D., Ph.D. (Harvard University) and Nathan S. Bryan, PhD (Texas), New York, NY, Springer Science and Business Media, Second Edition, In Press (book chapter). Huang L, Drake VJ, Ho E. Zinc. Adv Nutr. 2015 Mar 13;6(2):224-6. doi: 10.3945/an.114.006874. Jackson GS, Einstein JA, Kubley T, Martin BR, Weaver CM, Caffee MW. Biomedical graphite and CaF2 preparation and measurement at PRIME Lab.  Nucl Instr and Meth in Phys Res B.  361:358-362, 2015. Jacome-Sosa M, EJ Parks, RS Bruno, E Tasali, GF Lewis, BO Scheenman, TM Rains. (2016). Postprandial metabolism of macronutrients and cardiometabolic risk: recent developments, emerging concepts, and future directions. Adv Nutr, 7(2):364-74. Jakeman S, Weaver CM. Ch. 20  The effect of prebiotics on calcium absorption and utilization.  In:  Calcium:  Chemistry, Analysis, Function and Effects.  Preedy, VR, ed.  Royal Society of Chemistry  Pp  329-348, 2015. Kelly AM, Plautz SA, Zempleni J, Pannier AK. Glucocorticoid cell priming enhances transfection outcomes in adult human mesenchymal stem cells. Mol Ther. 2016;24(2):331-41. doi: 10.1038/mt.2015.195. Kim B, C. Farruggia, C. S. Ku, T. X. Pham, Y. Yang, C. J. Wegner, N. J. Farrell, E. Harness, Y. Park, S. I. Koo, J. Lee. Astaxanthin, a xanthophyll carotenoid, inhibited inflammation and fibrosis in the adipose and the liver of diet-induced obesity and liver fibrosis mice. J Nutr Biochem 2016; (In press). Epub ahead of print on March 2, 2016. Kim B, S. Lee, Y. Park, C. S. Ku, T. X. Pham, C. J. Wegner, Y. Yang, S. I. Koo, O. K. Chun, J. Lee. Blueberry, blackberry, and blackcurrant differentially affect plasma lipids and pro-inflammatory markers in diet-induced obesity mice. Nutr Res Pract 2016; (In revision). Klimis-Zacas D, Vendrame S and Kristo A, Wild blueberries attenuate risk factors of the Metabolic Syndrome, Journal of Berry Research, DOI:1.3233/JBR-160136, 2016 Koga, CC, Andrade, JE, Ferruzzi, MG, Lee, Y. Stability of trans-resveratrol encapsulated in a protein matrix produced using spray drying to UV light stress and simulated gastro-intestinal digestion. Journal of Food Science. 81(2): C292-C300 (2016) Kusuma RJ, Manca S, Friemel T, Sukreet S, Nguyen C, Zempleni J. Human vascular endothelial cells transport foreign exosomes from cow’s milk by endocytosis. Am J Physiol Cell Physiol. 2016:310(10):C800-7. doi: 10.1152/ajpcell.00169.2015 Lee, H, Dos Santos, L, Andrade, JE, Engeseth, N, Feng, H. Soy protein nano-aggregates with improved functional properties prepared by sequential pH treatment and ultrasonication. Food Hydrocolloids. 55:200-9. (2016) Li J, TN Sapper, E Mah, S Rudraiah, KE Schill, C Chitchumroonchokchai, MV Moller, JD McDonald, PR Rohrer, JE Manautou, RS Bruno (2016). Green tea extract provides extensive Nrf2-independent protection against lipid accumulation and NFκB pro-inflammatory responses during nonalcoholic steatohepatitis in mice fed a high-fat diet. Mol Nutr Food Res, 60(4):858-70. Mah E, TN Sapper, C Chitchumroonchokchai, ML Failla, KE Schill, SK Clinton, G Bobe, MG Traber, RS Bruno (2015). α-Tocopherol bioavailability is lower in adults with metabolic syndrome regardless of dairy fat co-ingestion: a randomized, double-blind, crossover trial. Amer J Clin Nutr, 102(5):1070-80. Martin BR, McCabe GP, McCabe L, Jackson GS, Horcajada MN, Offord-Cavin E, Peacock M, Weaver CM. Effect of Hesperidin with and without a calcium (Calcilock®) supplement on bone health in postmenopausal women.  J Clin Endocrinol Metab.  101:923-927, 2016. Molloy, A. M., Pangilinan, F., Mills, J. L., Shane, B., O’Neill, M. B., McGaughey, D. M., Velkova, A., Abaan, H. O., Ueland, P. M., McNulty, H., Ward, M., Strain, J. J., Cunningham, C., Casey, M., Cropp, C. D., Kim, Y., Bailey-Wilson, J. E., Wilson, A. F. and Brody, L. C. (2016) A common polymorphism in HIBCH influences methylmalonic acid concentrations in blood independently of cobalamin. Am. J. Hum. Genet, 98, 869–882. Moukahel, A-A., Asaad, S., Debbes, B., Morris, C.E.,  Sands, D.C. 2015. Characterization of wheat genotypes as sources of ice nucleation active bacteria for bioprecipitation aerosols. Int. J. Phytopathol. 04 (02) 2015. 63-72. Mullington JM, Abbott SM, Carroll JE, Dijk DJ, Dinges DF, Gehrman PR, Ginsburg GS, Gozal D, Haack M, Lim DC, Macrea M, Pack AI, Plante DT, Teske JA and Zee P. Developing Biomarker Assays Predicting Sleep and Circadian-Coupled Risks to Health. 2016. Sleep. April 1:39(4):727-36. Epub 2016 Feb. 29. Ojo B, El-Rassi GD, Perkins-Veazie P, Clarke S, Smith BJ, Lucas EA. Mango supplementation modulates gut microbial dysbiosis and short chain fatty acid production independent of body weight reduction in C57BL/6 mice fed a high fat diet. Journal of Nutrition. 2016 Jun 29. pii: jn226688. [Epub ahead of print] Opoku-Acheampong, A.B., Penugonda, K., Lindshield, B.L. Effect of Saw Palmetto Supplements on Androgen-Sensitive LNCaP Human Prostate Cancer Cell Number and Syrian Hamster Flank Organ Growth. Evid Based Complement Alternat Med. 2016: 8135135, 2016. doi:10.1155/2016/8135135 <a href="http://www.hindawi.com/journals/ecam/2016/8135135/">http://www.hindawi.com/journals/ecam/2016/8135135/</a> Phillips-Eakley AK, McKenney-Drake ML, Bahls M, Newcomer SC, Radcliffe JS, Wastney ME, Van Alstine WG, Jackson G, Alloosh M, Martin B, Sturek M, Weaver CM. Effect of high-calcium diet on coronary artery disease in Ossabaw miniature swine with metabolic syndrome.  J Am Heart Assoc  e001620, 2015. Rababah, T, Alhamad, M, Alu'datt, M, Al-mahasneh, M, Ereifej, K, Andrade, JE, Altarifi, B, Almajwal, A, Yang, W. The effect of drying process on total phenolics, antioxidant activity, and flavonoid contents of common Mediterranean herbs. International Journal of Agricultural and Biological Engineering. 8(5):145-50. (2015) Rajendran P, Dashwood WM, Li L, Kang Y, Kim E, Johnson G, Fischer KA, Löhr CV, Williams DE, Ho E, Yamamoto M, Lieberman DA, Dashwood RH. Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon. Clin Epigenetics. 2015 Sep 18;7(1):102. doi: 10.1186/s13148-015-0132-y Rendina-Ruedy E, Hembree K, Sasaki A, Davis M, Lighfoot SA, Clarke SL, Lucas EA, Smith BJ. A comparative study of the metabolic and skeletal response of C57BL/6J and C57BL/6N mice in a diet-induced model of type 2 diabetes. J Nutr Metab 2015 Shu J, Chiang K, Zempleni J, Cui J. Computational characterization of exogenous microRNAs that can be transferred into human circulation. PLoS ONE 10:e0140587, 2015 Sun Q, Y. Yue, P. Shen, J. J. Yang, and Y. Park (2016) Cranberry Bioactives Decrease Fat Accumulation in C. elegans, J. Med. Food. 19: 427-433. Teske JA, Perez-Leighton CE, Noble EE, Wang C, Billington CJ and Kotz CM. Effect of Housing Types on Growth, Feeding, Physical Activity and Anxiety-like Behavior in Male Sprague-Dawley Rats. 2016. Frontiers in Nutrition. 3:42016. <a href="http://dx.doi.org/10.3389/fnut.2016.00004.%20eCollection%202016">http://dx.doi.org/10.3389/fnut.2016.00004. eCollection 2016</a>. Teixeira DC, Cordonier EL, Wijeratne SSK, Huebbe P, Jamin A, Jarecke S, Wiebe M, Zempleni J. A cell death assay for assessing the mitochondrial targeting of proteins (submitted) Van Noordwijk M, Bruijnzeel S, Ellison D, Sheil D, Morris C, Sands D, Gutierrez V, Cohen J, Sullivan CA, Verbist B, Murdiyarso D, Gaveau D, Muys B. 2015. Ecological rainfall infrastructure: investment in trees for sustainable development. ASB Policy Brief 47. Nairobi: ASB Partnership for the Tropical Forest Margins. Vendrame S, Tsakiroglou P, Kristo A, Schuschke D, Klimis-Zacas D. Wild blueberry consumption attenuates local inflammation in the Perivascular Adipose Tissue of Obese Zucker Rats, Applied Physiology, Nutrition and Metabolism, accepted for publication, June, 2016 Watson GW, Wickramasekara S, Fang Y, Maier CS, Williams DE, Dashwood RH, Perez VI, Ho E. HDAC6 activity is not required for basal autophagic flux in metastatic prostate cancer cells. Exp Biol Med (Maywood). 2015 Dec 6. pii: 1535370215618518. [Epub ahead of print] Watson GW, Wickramasekara S, Fang, Y. Maier CS, Williams DE, Dashwood RH, Perez, VI, Ho E. Application of online RP/RP 2D LC-MS/MS method for the global proteomic profiling of prostate cancer cells. EuPA Open Proteom. 2015 Dec;9:34-40. Watson GW, Wickramasekara S, Maier CS, Williams DE, Dashwood RH, Ho E. (2015) Analysis of autophagic flux in response to sulforaphane in metastatic prostate cancer cells. Mol Nutr Food Res Oct;59(10):1954-61 Weaver CM. Parallels between nutrition and physical activity:  Research questions in development of peak bone mass.  Res Quart Exer Sports  86:103-106, 2015. Weaver CM, Alexander DD, Boushey CJ, Dawson-Hughes B, Lappe JM, LeBoff MS, Looker AC, Wallace TC, Wang DD. Calcium plus vitamin D supplementation and risk of fractures:  an updated meta-analysis from the National Osteoporosis Foundation.  Osteoporos Int DOI 10.1007/s00198-015-3386-5. Weaver CM, Jakeman S. Ch. 14 Prebiotics, calcium absorption, and bone health:  In:  Nutritional Influences of Bone Health.  International Congress Series Proceedings of the 9th International Symposium on Nutrition Aspects of Osteoporosis, Montreal Canada.  Weaver CM, Daly R, Bischoff-Ferrari H, eds, Springer, 2016, in press. Weaver CM, Lawlor M, McCabe GP. Ch. 16 Predicting calcium requirements in children.  In:  Nutritional Influences of Bone Health.  International Congress Series Proceedings of the 9th International Symposium on Nutrition Aspects of Osteoporosis, Montreal Canada.  Weaver CM, Daly R, Bischoff-Ferrari H, eds, Springer, 2016, in press. Whisner CM, Martin BR, Nakatsu, CH, Story JA, MacDonald-Clarke CJ, McCabe LD, McCabe GP, Weaver CM. Soluble corn fiber increases calcium absorption associated with shifts in the gut microbiome.  A randomized dose-response trial in free-living pubertal girls.  J Nutr, 2016;146(7):1298-306.  doi: 10.3945/jn.115.227256 Wolf T, Baier SR, Zempleni J. The intestinal transport of bovine milk exosomes is mediated by endocytosis in human colon carcinoma Caco-2 cells and rat small intestinal IEC-6 cells. J Nutr 145:2201-2206, 2015 [Teresa A. Davis, Editor-in-Chief of The Journal of Nutrition, has selected this article as the Editor’s Pick for Volume 145 Issue 10 of the journal. The article was featured on the journal website at <a href="http://jn.nutrition.org/">http://jn.nutrition.org</a> , and in the member newsletter.] Yang Y, M. Bae, B. Kim, Y. Park, S. I. Koo, J. Lee. Astaxanthin prevents and reverses the activation of mouse primary hepatic stellate cells. J Nutr Biochem 2016; 29:21-26. Epub ahead of print on Nov 24, 2015. Yang Y, M. Bae, Y. Park, Y. Lee, T. X. Pham, S. Rudraiah, J. Manautou, S. I. Koo, J. Lee. Astaxanthin prevents and reverses the activation of hepatic stellate cells via the modulation of histone deacetylase 9. Mol Nutr Food Res 2016; (In review). Zempleni J, Baier SR, Hirschi K. Diet-responsive microRNAs are likely exogenous [letter to the editor] J Biol Chem 289:25197, 2015 Zempleni J, Baier SR, Howard KM, Cui J. Gene regulation by dietary microRNAs. In: Nutrients/natural product (nutraceutical) control of metabolic pathways in relation to the metabolic syndrome (Dakshinamurti, K, Zempleni J, guest editors). Can J Physiol Pharmacol 93:1097-1102, 2015

Impact Statements

Dietary bioactive ingredients like prebiotics are beneficial to bone and help protect against shortfall minerals in the diet, especially calcium. Purdue University, Connie Weaver

Date of Annual Report: 06/19/2017

Report Information

Annual Meeting Dates: 05/23/2017 - 05/24/2017
Period the Report Covers: 10/01/2015 - 09/30/2016

Participants

Andrade, Juan (jandrade@illinois.edu) – University of Illinois, Urbana-Champaign
Bruno, Richard (Bruno.27@osu.edu) – Ohio State University
Ho, Emily (emily.ho@oregonstate.edu) – Oregon State University
Hord, Norm (Norman.Hord@oregonstate.edu) - Oregon State University
Lindshield, Brian (Chair, blindsh@ksu.edu) - Kansas State University
Liu, Yanhong (yahliu@ucdavis.edu) - University of California, Davis
Park, Yeonhwa (ypark@foodsci.umass.edu) – University of Massachusetts
Sands, David (uplds@montana.edu) – Montana State University
Shane, Barry (bandie@berkeley.edu) University of California – Berkeley
Shipka, Milan (Administrative Advisor, mpshipka@alaska.edu) – University of Alaska, Fairbanks
Teske, Jennifer (Secretary, teskeja@email.arizona.edu) - University of Arizona
Weaver, Connie (weavercm@purdue.edu) - Purdue
Zempleni, Janos (jzempleni2@unl.edu) - University of Nebraska, Lincoln
Cotton, Paul (USDA NIFA National Program Leader, Pau.Cotton@nifa.usda.gov via Skype)

Brief Summary of Minutes

Meeting was called to order on May 23 and May 24, 2017 at 8:15 a.m. PST at the Oregon State University Portland Center.

Welcome and Introductions (May 23) – Participants were welcomed by the host (Emily Ho, Oregon State University) and Meeting Chair, Dr. Brian Lindshield, Kansas State University.

Executive session (May 24) – Dr. Jennifer Teske was elected by the members present to be the chair of the next annual meeting. She will coordinate next year’s W3002 meeting in Tucson, Arizona; targeted for some time in February 2017. Yanhong Liu, University of California, Davis, was elected as secretary.

Dr. Lindshield described the meeting objectives, timeline, and responsibilities.

Dr. Paul Cotton, NIFA Program Leader, joined the meeting via Skype and provided an update on the NIFA budget and current and future funding opportunities on May 24.

Each W3002 Investigator attending the meeting provided a brief oral progress report. At the end of each presentation a short discussion took place.

The group focused their attention on discussion and writing for the upcoming renewal this year. Drs. Ho. and Lindshield lead the discussion to first determine research priorities and membership for the upcoming renewal. Then the group assembled three writing teams and each group was modified the grant based on the pre-determined research priorities. Drs. Ho. and Lindshield brought the group together to reflect and clarify sections within the grant to ensure the group was agreed on work performed by each group. The group continued in this manner for the reminder of the meeting. The final group discussion ended with each member tasked with specific writing assignments.

Our new administrative advisor, Dr. Milan Shipka, introduced himself and provided guidance on the project’s renewal.

Included in the agenda was a tour of the Food Innovation Lab.

The meeting adjourned on May 24, 2016 at 4:30 p.m. An informal dinner took place after the meeting.

Accomplishments

Publications

<ul> <li>Abbas, A., J. A. Hall, W. L. Patterson, 3rd, E. Ho, A. Hsu, F. Al-Mulla and P. T. Georgel (2016). "Sulforaphane modulates telomerase activity via epigenetic regulation in prostate cancer cell lines." Biochem Cell Biol 94(1): 71-81.</li> <li>Bailey RL, Weaver CM, Murphy S. Using the Dietary Reference Intakes to assess intakes in Research:  Successful Approaches.  Van Horn L, ed.  Academy of Nutrition and Dietetics, Chicago IL, 2017.</li> <li>Brannon PM, Weaver CM, Anderson CAM, Donovan SM, Murphy SP, Yaktine AL. Scanning for new evidence to prioritize updates to the Dietary Reference Intakes:  case studies for thiamin and phosphorus.  Am J Clin Nutr  104:1-12, 2016.</li> <li>Beaver, L. M., R. Kuintzle, A. Buchanan, M. W. Wiley, S. T. Glasser, C. P. Wong, G. S. Johnson, J. H. Chang, C. V. Lohr, D. E. Williams, R. H. Dashwood, D. A. Hendrix and E. Ho (2017). "Long noncoding RNAs and sulforaphane: a target for chemoprevention and suppression of prostate cancer." J Nutr Biochem 42: 72-83.</li> <li>Choi M-K, Weaver, CM. Daily intake of magnesium and its relation to urinary excretion in Korean healthy adults consuming self-selected diets.  Biol Trace Elem Res.  DOI:  1007/s12011-016-0822-0.</li> <li>Conley MN, Roberts C, Sharpton TJ, Iwaniec UT, Hord NG. (2017) Increasing dietary nitrate has no effect on cancellous bone loss or fecal microbiome in ovariectomized rats. Mol Nutr Food Res. May;61(5). doi: 10.1002/mnfr.201600372. Epub 2017 Mar 30.</li> <li>Conley, M and Hord, NG (2017) Regulation of dietary nitrate and nitrite: balancing essential physiological roles with potential health risks, In: Nitrates and Nitrites in Human Health and Disease, Editors: Joseph Loscalzo, M.D., Ph.D. (Harvard University) and Nathan S. Bryan, PhD (Texas), New York, NY, Springer/Humana Press, 2nd edition (book chapter).</li> <li>Conley MN, Wong CP, Duyck KM, Hord NG, Ho E, Sharpton TJ. (2016) <a href="https://www.ncbi.nlm.nih.gov/pubmed/27069796">Aging and serum MCP-1 are associated with gut microbiome composition in a murine model.</a> 2016 Mar 31;4:e1854. doi: 10.7717/peerj.1854.</li> <li>Dain A, Repossi D, Diaz-Gerevini GT, Vanamala J, Das UN, Eynard AR. 2016. Effect of polyunsaturated fatty acids with and without nordihydroguaiaretic acid (NDGA) on metabolic and inflammatory indices in a rat model of spontaneous type 2 diabetes mellitus. Lipids in Health and Disease. </li> <li>Del Bo C, Porrini M, Campolo J, Parolini M, Lanti C, Klimis-Zacas D, Riso P 2016. A single blueberry (Vaccinium corymbosum) portion does not affect markers of antioxidant defense and oxidative stress in healthy volunteers following cigarette smoking, Mutagenesis 31:215-214.</li> <li>Bobe G, CobbTJ, Leonard SW, Aponso S, Bahro CB, Koley D, Mah E, Bruno RS, Traber MG (2017). Increased static and decreased capacity oxidation-reduction potentials in plasma are predictive of metabolic syndrome. Redox Biol, 12:121-128.</li> <li>Bukenya, R, Ahmed, A, Andrade, JM, Grigsby-Toussaint, DS, Muyonga, J, Andrade, JE. Validity and reliability of general nutrition knowledge questionnaire for adults in Uganda. Nutrients. 9(2): 172. (2017)</li> <li>Charepalli V*, Reddivari L, Vadde R*, Walia S, Radhakrishnan S*, Vanamala J. 2016. Eugenia jambolana (Java Plum) fruit extract exhibits anti-cancer activity against early stage human HCT-116 colon cancer cells and colon cancer stem cells. Cancers 8(3):pii E29. </li> <li>Coborn JA, Deporter DP, Mavanji, V, Sinton CM, Billington CJ, Kotz CM, Teske, JA. Role of orexin A in the ventrolateral preoptic area on components of total energy expenditure. International Journal of Obesity. Accepted March 27, 2017. (in press) Epub 4/24/17.</li> <li>Conley, M. N., C. P. Wong, K. M. Duyck, N. Hord, E. Ho and T. J. Sharpton (2016). "Aging and serum MCP-1 are associated with gut microbiome composition in a murine model." PeerJ 4: e1854.</li> <li>Cordonier EL, Jarecke SK, Hollinger FE, Zempleni J. Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells. Eur J Pharmacol 780:202-208, 2016Cotten BM, Diamond SD, Banh T, Hsiao Y-H, Cole RM, Li J, Simons CT, Bruno RS, Belury MA, Vodovotz Y. (2017). Raspberry ketone fails to reduce adiposity beyond decreasing food intake in C57BL/6 mice fed a high-fat diet. Food Funct, 8(4):1512-1518</li> <li>Deac OM, Mills JL, Gardiner CM, Shane B, Quinn L, Midttun Ø, McCann A, Meyer K, Ueland PM, Fan R, Lu Z, Brody LC, Molloy AM. (2016) Serum immune system biomarkers neopterin and interleukin-10 are strongly related to tryptophan metabolism in healthy young adults. Nutr. doi: 146: 1801-1806.</li> <li>Del Bo C, Deon V, Campolo J, Porrini M, Klimis-Zacas D, Riso P. A serving of blueberry (V. corymbosum) reverses endothelial dysfunction in young smokers and non-smokers: a randomized, controlled, crossover study, Food and Function, 2017 (invited paper, submitted)</li> <li>Delimont NM, Chanadang S, Joseph MV, Rockler BE, Guo Q, Regier GK, Mulford MR, Kayanda R, Range M, Mziray Z, Ambaksye J, Mugyabuso J, Msuya W, Lilja NK, Procter SB, Chambers IV E, Alavi S, Lindshield BL. The MFFAPP Tanzania efficacy study protocol: newly formulated, extruded fortified-blended foods for food aid. Curr Dev Nutr. 2017: 1(4) doi: <a href="https://doi.org/10.3945/cdn.116.000315">https://doi.org/10.3945/cdn.116.000315</a>.</li> <li>Delimont NM, Fiorentino NM, Opoku-Acheampong AB, Joseph MV, Guo Q, Alavi S, Lindshield BL. Newly formulated, extruded sorghum, cowpea, corn, and soy containing fortified-blended foods lead to adequate vitamin A, iron outcomes and improved growth compared with CSB+ in rats. J Nutr Sci. 6: e18 doi:10.1017/jns.2017.15.</li> <li>Delimont NM, Haub M, Lindshield BL. Does Tannin Consumption Decrease Iron Bioavailability? Curr Dev in Nutr. 2017. 1(2) doi: <a href="https://doi.org/10.3945/cdn.116.000042">https://doi.org/10.3945/cdn.116.000042</a>.</li> <li>Ellison D, Morris CE, Locatelli B, Sheil D, Cohen J, Murdiyarso D, Gutierrez V, van Noordwijk M, Creed IF, Pokorny J, Gaveau D, Spracklen D, Bargués Tobella A, Ilstedt U, Teuling R, Gebrehiwot SG, Sands DC, Muys B, Verbist B, Springgay E, Sugandi Y, Sullivan C. 2017. Trees, forests and water: cool insights for a hot world. Global Environmental Change (in press)</li> <li>Ferira AJ, Laing EM, Hausman DB, Hall DB, McCabe GP, Martin BR, Hill KM, Warden SJ, Weaver CM, Peacock M, Lewis RD. Vitamin D supplementation effects on insulin sensitivity and resistance in early pubertal white and black children.  J Clin Endocrinol Metab, In Press, 2016.</li> <li>Fernández-Aparicio M, Bernard A, Falchetto L, Margert P, Chauvel B, Steinberg C, Morris CE, Gibot-Leclerc S, Boari A, Vurro M, Bohan DA, Sands DC, Reboud X. (2017 Investigation of amino acids as herbicides for control of Orobanche minor parasitism in red clover. Frontiers in Plant Science vol. 8, May 22, 2017; article 842.</li> <li>Gaur, S, Sloffer, EM, Ojha, A, Patra, F, Shukla, S, Engeseth, NJ, Patel, PR, Andrade, JE. Omega-3 fortified lipid-based nutrient supplement – development, characterization and consumer acceptability. Food and Nutrition Bulletin. (2017). [In press]</li> <li>Graef J, Rendina-Ruedy E, Crockett E, Ouyang P, King J, Cichewicz RH, Lucas EA, Smith BJ. Osteoclast differentiation is downregulated by select polyphenolic fractions from dried plum via suppression of MAPKs and Nfatc1. Plos ONE (under review).</li> <li>Guo X, Wu L, Wang W, Medeiros DM, Clarke S, Lucas E, Smith BJ, Chowanadisai W, Lin D. Hypothalamic mitochondria in energy homeostasis and obesity. Integ Mol Med. (in press)</li> <li>Guo X, Wu L, Lyu Y, Chowanadisai W, Clarke SL, Lucas EA, Smith BJ, He H, Wang W, Medeiros DM, Lin D. <a href="https://www.ncbi.nlm.nih.gov/pubmed/28482236">Ablation of β,β-carotene-9',10'-oxygenase 2 remodels the hypothalamic metabolome leading to metabolic disorders in mice.</a> J Nutr Biochem. 2017;46:74-82.</li> <li>Jacome-Sosa M, Parks EJ, Bruno RS, Tasali E, Lewis GF, Scheenman BO, Rains TM. (2016). Postprandial metabolism of macronutrients and cardiometabolic risk: recent developments, emerging concepts, and future directions. Adv Nutr, 7(2):364-74.</li> <li>Jakeman SA, Henry CN, Martin BR, McCabe GP, McCabe LD, Jackson JS, Peacock M, Weaver CM. Soluble corn fiber increases bone retention in postmenopausal women in a dose-dependent manner:  a randomized crossover trial.  Am J Clin Nutr 104:837-843, 2016.Jiang, S, Ding, J, Andrade, JE, Rababah, TM, Almajwal A, Abulmeaty, MM, Feng, H.  Modifying the physicochemical properties of pea protein by pH-shifting and ultrasound combined treatments. Ultrasonics Sonochemistry. [In Press]</li> <li>Johnson, G. S., J. Li, L. M. Beaver, W. M. Dashwood, D. Sun, P. Rajendran, D. E. Williams, E. Ho and R. H. Dashwood (2016). "A functional pseudogene, NMRAL2P, is regulated by Nrf2 and serves as a coactivator of NQO1 in sulforaphane-treated colon cancer cells." Mol Nutr Food Res.</li> <li>Kim K, Ehrlich A, Perng V, Chase J, Raybould H, Li X, Atwill ER, Whelan R, Sokale A, Liu Y. 2017. Effects of dietary β-glucan on growth performance, diarrhea, and gut permeability of weanling pigs experimentally infected with a pathogenic coli. ASAS-CSAS Annual Meeting & Trade Show.</li> <li>Kim K, Perng V, Chase J, Li X, Atwill ER, Whelan R, Sokale A, Liu Y. 2017. Effects of dietary β-glucan on systemic immunity of weanling pigs experimentally infected with a pathogenic  coli. <a href="http://animalnutr-ansci.faculty.ucdavis.edu/abstracts/2017-canc-conference_digital/">California Animal Nutrition Conference Proceedings</a>. p.282.</li> <li>Kopecky SL, Bauer DC, Gulati M, Nieves JW, Singer AJ, Toth PP, Underberg JA, Wallace, TC, Weaver CM. Lack of evidence linking calcium with or without vitamin D supplementation to cardiovascular disease in generally healthy adults: A position statement from The National Osteoporosis Foundation and American Society for Preventive Cardiology  Ann Intern Med  2016  DOI:10.7326/M16-1743.</li> <li>Kristo A, Klimis-Zacas D, Sikalidis AK, Protective role of dietary berries in cancer, In Antioxidants (Special Issue, Berries in Health and Disease, guest editor Dorothy Klimis-Zacas), 2016, 5(4), 37; doi:<a href="http://dx.doi.org/10.3390/antiox5040037">3390/antiox5040037</a></li> <li>Lee J, Yue Y, Park Y, Lee S-H, 3,3'-Diindolylmethane Suppresses Adipogenesis via AMPK-Dependent Mechanism in 3T3-L1 Adipocytes and Caenorhabditis elegans, Med. Food (Accepted)</li> <li>Li J, Sapper TN, Mah E, Moller MV, Kim JB, Chitchumroonchokchai C, McDonald JD, Bruno RS. (2017). Green tea extract treatment reduces NFκB activation in mice with diet-induced nonalcoholic steatohepatitis by lowering TNFR1 and TLR4 expression and ligand availability. J Nutr Biochem, 41:34-41.</li> <li>Li Y, Bharath LP, Qian Y, Ruan T, Babu PVA, Bruno RS, Symons JD, Jalili T (2016). γ-Carboxyethyl hydroxychroman, a metabolite of γ-tocopherol, preserves nitric oxide bioavailability in endothelial cells challenged with high glucose. Exp Biol Med, 241(18):2056-2062.</li> <li>Lipkie T, Ferruzzi M, Weaver C. Low bioaccessibility of vitamin D2 from yeast fortified bread compared to crystalline D2 bread and D3 from fluid milks.  Food & Function </li> <li>Liu Y. 2016. Non-nutrients in swine health and production. J. Anim. Sci. 94 (E-Suppl.5):489.</li> <li>Liu Y, Gonzalez-Vega JC, Vazquez-Anon M, Zhao J, Escobar J, Almeida FN, Stein HH. 2016. Effects of phytase on growth performance and metacarpal bone ash in weanling pigs. J. Anim. Sci. 94 (E-Suppl.2):111.</li> <li>Kusuma JR, Manca S, Friemel T, Sukreet S, Nguyen C, Zempleni J. Human vascular endothelial cells transport foreign exosomes from cow’s milk by endocytosis. Am J Physiol Cell Physiol 310:C800-C807, 2016</li> <li>Pannier A, Kelly A, Plautz S, Zempleni J. Glucocorticoid cell priming enhances transfection outcomes in adult human mesenchymal stem cells. Mol Ther 24:331-341, 2016</li> <li>Parrish JB and Teske JA. Acute partial sleep deprivation due to environmental noise increases weight gain by reducing energy expenditure in rodents. 2017. Obesity (Silver Spring) Jan;25(1):141-146. Epub 11/29/16.</li> <li>Massey AR*, Reddivari L, Radhakrishnan S*, Karunathilake E*, Vanamala J. 2016. Pro-apoptotic activity against human cancer stem cells differs between different parts of sweet sorghum, a feedstock for biorefining. Journal of Functional Foods. 25: 267–278. </li> <li>Macdonald-Clark, CJ, Martin BR, McCabe LD, McCabe GP, Lachcik PJ, Wastney M, Weaver CM. Bioavailability of potassium from potatoes and potassium gluconate:  a randomized dose response trial.  Am J Clin Nutr  104:346-353, 2016.</li> <li>Morris CE, Sands DC. 2017. Impacts of microbial aerosols on natural and agro-ecosystems: immigration, invasions and their consequences. IN: Delort A.M., Amato, P. (eds.) Microbiology of Bioaerosols. Wiley, (in press).</li> <li>Morris CE, Soubeyrand S, Bigg EK, Creamean JM, Sands DC. 2016. Mapping rainfall feedback to reveal the potential sensitivity of precipitation to biological aerosols. Bull. Amer. Meteorol. Soc. doi.org/10.1175/BAMS-D-15-00293.1</li> <li>Nzioki HS, Oyosi F, Morris C, Kaya E, Pilgeram A, Baker C, Sands D.C. 2016, Striga biocontrol on a toothpick: A readily deployable and inexpensive method for smallholder farmers. Frontiers in Plant Sci. 7: Article 1121.</li> <li>Ojo B, Simenson AJ, O’Hara C, Wu L, Gou X, Peterson SK, Lin D, Smith BJ, Lucas EA. Wheat germ supplementation alleviates insulin resistance and cardiac mitochondrial dysfunction in an animal model of diet-induced obesity. Br J Nutr (under review)</li> <li>Ojo B, Davila El-Rassi G, Payton M, Perkins-Veazie P, Clarke S, Smith BJ, Lucas EA. Mango supplementation prevents gut microbial dysbiosis and modulates short chain fatty acid production independent of body weight reduction in C57BL/6 mice fed a high fat diet. J Nutr. 2016;146(8):1483-91.</li> <li>Pinto JT, Zempleni J. Nutrition information brief – riboflavin. Adv Nutr 7:973-975, 2016Pham, TX, Park Y, Bae M, Lee J. The potential role of an endotoxin-like mechanism for the anti-inflammatory effect of Spirulina platensis in macrophages: Insight into energy phenotype. J Med Food 2017; 20: 201-210.</li> <li>Pham TX, Park Y-K, Lee J. Anti-inflammatory effects of Spirulina platensis extract via the modulation of histone deacetylases. Nutrients 2016; 8:E381.</li> <li>Pham TX, Lee J. Anti-inflammatory effect of Spirulina platensis in macrophages is beneficial for adipocyte differentiation and maturation by inhibiting Nuclear factor-kB pathway in 3T3-L1 adipocytes. J Med Food 2016; 19:535-542.</li> <li>Reddivari L, Charepalli V*, Radhakrishnan S*, Vadde R*, Elias R, Lambert J, Vanamala J. 2016. Dietary grape compounds suppress oncogenic stem cells in a mouse model of chemically-induced colon cancer. BMC Complementary and Alternative Medicine. 16:278-290. </li> <li>Rendina-Ruedy E, Graef JL, Lightfoot SA, Ritchey JW, Clarke SL, Lucas EA, Smith BJ. Impaired glucose intolerance attenuates bone accrual in young growing skeleton by promoting the maturation of osteoblasts: role of beclin1-mediated autophagy. Bone Reports 2016 5:199-207.</li> <li>Sapper TN, Mah E, Ahn-Jarvis J, McDonald JD, Chitchumroonchokchai C, Reverri EJ, Vodovotz Y, Bruno RS. (2016). A green tea-containing starch confection increases plasma catechins without protecting against postprandial impairments in vascular function in normoglycemic adults. Food Funct, 7:3843-3853.</li> <li>Shams-White MM, Chung M, Du M, Fu Z, Insogna KL, Karlsen MC, LeBoff MS, Shapses SA, Sackey J, Wallace TC, Weaver CM. Dietary protein and bone health:  a systematic view and meta-analysis from the National Osteoporosis Foundation.  2017  3945/ajcn.116.145110.</li> <li>Sharif S, Mustafa G, Munir H, Weaver CM, Jamil Y, Shahid. Proximate composition and micronutrient mineral profile of wild Ganoderma lucidium and four commercial exotic mushrooms by ICP-OES and LIBS.  J Food Nutr Res  4:703-708, 2016.</li> <li>Shen P, Yue Y, Kim K-H, ParkY Piceatannol Reduces Fat Accumulation in Caenorhabditis elegans, Med. Food (Accepted)</li> <li>Shen P, Yue Y, Sun Q, Kasireddy N, Kim K-H, Park Y Piceatannol Extends the Lifespan of Caenorhabditis elegans Via DAF-16, Biofactors. (Accepted)</li> <li>Smith BJ, Crockett EK, Chongwatpol P, Graef JL, Rendina-Ruedy E, Clarke SL, Lucas EA. Tart cherry supplementation demonstrates anabolic properties by increasing bone mineralization in a model of age-related bone loss. J Nutr (under review).</li> <li>Song M, Charoensinphon N, Wu X, Zheng J, Gao Z, Xu F, Wang M, Xiao H (2016) Inhibitory effects of metabolites of 5-demethylnobiletin on human nonsmall cell lung cancer cells, J. Ag. Food Chem. 64: 4943-4949.</li> <li>Teixeira CD, Cordonier EL, Wijeratne SSK, Huebbe P, Jamin A, Jarecke S, Wiebe M, Zempleni J. A cell death assay for assessing the mitochondrial targeting of proteins (submitted)</li> <li>Thorning TK, Bertram HC, Bonjour JP, de Groot L, Dupont D, Feeney E, Ipsen R, Lecerf JM, Mackie A, McKinley MC, Michalski MC, Rémond D, Risérus U, Soedamah-Muthu SS, Tholstrup T, Weaver C, Astrup A, Givens I. Whole dairy matrix or single nutrients in assessment of health effects: current evidence and knowledge gaps.  Am J Clin Nutr  doi: 10.3945/ajcn.116.151548.</li> <li>Thomson, C. A., E. Ho and M. B. Strom (2016). "Chemopreventive properties of 3,3'-diindolylmethane in breast cancer: evidence from experimental and human studies." Nutr Rev 74(7): 432-443.</li> <li>Traber MG, Mah E, Leonard SW, Bobe G, Bruno RS. (2017). Metabolic syndrome increases dietary α-tocopherol requirements as assessed using urinary and plasma vitamin E catabolites: a double-blind, crossover clinical trial. Amer J Clin Nutr, 105(3):571-579.</li> <li>Tsakiroglou P, Ashworth S, Webber J, DelBo C, Klimis-Zacas D, Anthocyanins and Phenolic acids extracted from wild blueberries (V. augustifolium) differentially modulate endothelial cell migration, Food and Function, 2017 (invited paper, submitted).</li> <li>Vadde R*, Radhakrishnan S*, Karunathilake E*, Reddivari L, Vanamala, J. 2016. Indian gooseberry (Emblica officinalis Gaertn.) suppresses cell proliferation and induces apoptosis in human colon cancer stem cells independent of p53 status via suppression of c-Myc and cyclin D1. Journal of Functional Foods. 23:601-613. </li> <li>Vendrame S, Tsakiroglou P, Kristo A, Schuschke D and Klimis-Zacas D, Wild blueberry consumption attenuates local inflammation in the Perivascular Adipose Tissue of Obese Zucker Rats, Applied Physiology, Nutrition and Metabolism, in press, 1–7 (0000) dx.doi.org/10.1139/apnm-2016-0160</li> <li>Vendrame S, Del Bo C, Ciappellano S, Riso P, Klimis-Zacas D, Berry fruit consumption and Metabolic Syndrome, In Antioxidants (Special Issue, Berries in Health and Disease, guest editor Dorothy Klimis-Zacas),2016, 5(4), 34; doi:<a href="http://dx.doi.org/10.3390/antiox5040034">3390/antiox5040034</a></li> <li>Vogel KA, Martin BR, McCabe LD, Peacock M, Warden SJ, McCabe GP, Weaver Cm. The effect of dairy intake on bone mass and body composition in early pubertal girls and boys:  A randomized controlled trial.  AJCN  105:1214-1229, 2017.</li> <li>Wallace TC, Marzorati M, Spence L, Weaver CM, Williamson PS. New frontiers in fibers:  Innovative and emerging research on the gut microbiome and bone health.  J Am Coll Nutr  2017</li> <li>Wang, R., Y. Kang, C. V. Lohr, K. A. Fischer, C. S. Bradford, G. Johnson, W. M. Dashwood, D. E. Williams, E. Ho and R. H. Dashwood (2016). "Reciprocal regulation of BMF and BIRC5 (Survivin) linked to Eomes overexpression in colorectal cancer." Cancer Lett 381(2): 341-348.</li> <li>Wang W, Yang J, Qi W, Yang H, Wang C, Tan B, Hammock BD, Park Y, Kim D, Zhang G (2017) Lipidomic Profiling of High-Fat Diet-Induced Obesity in Mice: Importance of Cytochrome P450-Derived Fatty Acid Epoxides, Obesity 25: 132-140.</li> <li>Watson, G. W., S. Wickramasekara, Y. Fang, C. S. Maier, D. E. Williams, R. H. Dashwood, V. I. Perez and E. Ho (2016). "HDAC6 activity is not required for basal autophagic flux in metastatic prostate cancer cells." Exp Biol Med (Maywood) 241(11): 1177-1185.</li> <li>Weaver CM. Ensuring adequate calcium without concern for safety.  Nutr Today  52:90-92, 2017.</li> <li>Weaver CM. Miracle berries:  how blueberries can improve bone health.  K Biobank - Research Publishing International  researchfeatures.com pg 64-67, 2016.</li> <li>Weaver CM, Hill Gallant KM 44 Osteoporosis:  The Early Years.  In:  Nutrition in the Prevention and Treatment of Disease  2017, 4th Ed.  Coulston, AM, Boushey, CJ, Ferruzzi MG, DeLahanty LM, eds.  Elsevier, Inc.  Pp 969-989.</li> <li>Weaver CM, Jakeman S. 14  Prebiotics, calcium absorption, and bone health:  In:  Nutritional Influences of Bone Health.  International Congress Series Proceedings of the 9th International Symposium on Nutrition Aspects of Osteoporosis, Montreal Canada.  Weaver CM, Daly R, Bischoff-Ferrari H,  eds, Springer, pgs 145-152, 2016.</li> <li>Weaver CM, Lawlor M, McCabe GP. 16  Predicting calcium requirements in children.  In:  Nutritional Influences of Bone Health.  International Congress Series Proceedings of the 9th International Symposium on Nutrition Aspects of Osteoporosis, Montreal Canada.  Weaver CM, Daly R, Bischoff-Ferrari H,  eds, Springer, pgs 171-178, 2016.</li> <li>Weaver CM, Martin, BR, McCabe GP, McCabe LD, Woodward M, Anderson CAM, Appel LJ. Individual variation in urinary sodium excretion among adolescent girls on a fixed intake.  J Hyperten  34:1290-1297, 2016</li> <li>Whisner CM, Martin BR, Nakatsu CH, Story JA, Macdonald-Clark CJ, McCabe LD, McCabe GP, Weaver CM. Soluble corn fiber increases calcium absorption associated with shifts in the gut microbiome:  A randomized dose-response trial in free-living pubertal females.  J Nutr  146:1298-1306, 2016.</li> <li>Wu L, Guo X, Hartson S, Davis A, He H, Medeiros D, Wang W, Clarke S, Lucas E, B Smith, von Lintig J, Lin D. Lack of β, β-carotene -9’, 10’-oxygenase (BCO2) leads to hepatic mitochondrial dysfunction and intracellular oxidative stress in mice. Mol Nutr Food Res. 2017;61(5). Epub 2017 Feb 9.</li> <li>Wu L, Guo X, Wang W, Medeiros DM, Clarke SL, Lucas EA, Smith BJ, Lin D. Molecular aspects of β, β-carotene-9’, 10’-oxygenase 2 in carotenoid metabolism and diseases. Exp Biol Med. (Maywood) 2016; 241(17):1879-1887.</li> <li>Yeoh BS, Saha P, Singh V, Xiao X, Ying Y, Vanamala J, Kennett M, Harvatine K, Joe B, and Vijay-Kumar M. Stearoyl CoA Desaturase-1 Aggravates Colitogenic Potential of Adoptively Transferred Effector T cells. American Journal of Physiology - Gastrointestinal and Liver Physiology. Accepted 6 September 2016. </li> <li>Yue JT , Abraham MA , Bauer PV , LaPierre MP , Wang P , Duca FA , Filippi BM , Chan O , Lam TK . Inhibition of glycine transporter-1 in the dorsal vagal complex improves metabolic homeostasis in diabetes and obesity. Nat Commun . 2016;7:13501.</li> <li>Zempleni J, Aguilar-Lozano A, Sadri M, Manca S, Wu D, Zhou F, Mutai E, Sukreet S. Biological activities of extracellular vesicles and their cargos from bovine and human milk in humans and implications for infants. J Nutr 147:3-10, 2017</li> <li>Zempleni J. Milk exosomes: beyond dietary microRNAs. Nutr Genes (in press)</li> <li>Zhang, Z., L. L. Atwell, P. E. Farris, E. Ho and J. Shannon (2016). "Associations between cruciferous vegetable intake and selected biomarkers among women scheduled for breast biopsies." Public Health Nutr 19(7): 1288-1295</li> <li>Zhong F, Xu M, Bruno RS, Ballard KD, Zhu J. (2017). Targeted high performance liquid chromatography tandem mass spectrometry-based metabolomics differentiates metabolic syndrome from obesity. Exp Biol Med, 242(7):773-780.</li> </ul>

Impact Statements

1. We have discovered a novel class of bioactive compounds in foods, i.e., exosomes and their RNA cargos. This research has major implications for the U.S. dairy and egg industry, and the way we assess the nutritional value of foods. External grant support totaling $2.3 million direct costs per year were secured. External funding and an appealing research program were leveraged to create opportunities for workforce development and student education. University of Nebraska, Lincoln (Janos Zempleni).

Date of Annual Report: 04/02/2018

Report Information

Annual Meeting Dates: 02/08/2018 - 02/09/2018
Period the Report Covers: 10/01/2017 - 09/30/2018

Participants

Bruno, Richard (Bruno.27@osu.edu) – Ohio State University, Columbus, OH
Dallas, David (dave.dallas@oregonstate.edu) - Oregon State University, Corvallis, OR
Duca, Frank (faduca@email.arizona.edu) – University of Arizona, Tucson, AZ
Gruen, Ingolf (grueni@missouri.edu) - University of Missouri, Columbia, MO
Hord, Norman (Norman.Hord@oregonstate.edu) - Oregon State University, Corvallis, OR
Lee, Ji-Young (ji-young.lee@uconn.edu) – University of Connecticut, Storrs, CT
Liu, Yanhong (yahliu@ucdavis.edu) - University of California, Davis, CA
Sands, David (uplds@montana.edu) – Montana State University, Bozeman, Montana
Shane, Barry (bandie@berkeley.edu) University of California, Berkeley, CA
Shipka, Milan (Administrative Advisor, mpshipka@alaska.edu) – University of Alaska, Fairbanks, AK
Teske, Jennifer (teskeja@email.arizona.edu) - University of Arizona, Tucson, AZ
Weaver, Connie (weavercm@purdue.edu) – Purdue University, West Lafayette, IN
White, Wendy (wswhite@iastate.edu) – Iowa State University, Ames, IA
Zempleni, Janos (jzempleni2@unl.edu) - University of Nebraska, Lincoln, NE

Brief Summary of Minutes

Meeting was called to order on Feb. 8 and Feb. 9, 2018 at 8:30 a.m. MST at the University of Arizona.

Welcome and Introductions (Feb. 8) – Participants were welcomed by the host and Meeting Chair (Jennifer Teske, University of Arizona).

Executive session (Feb. 8) – Dr. Yanhong Liu was elected by the members present to be the chair of the next annual meeting, 2019. She will coordinate next year’s W4002 meeting in Davis, CA; targeted for some time in February 2019. Dr. Richard Bruno, Ohio State University, was elected as secretary.

Dr. Milan Shipka, our administrative advisor, joined the meeting, updated the project’s renewal, and gave the guidance for annual report submission.

Each W3002 Investigator attending the meeting gave an oral progress report in the following order:

Bruno Richard, Ohio State University – “Anti-inflammatory mechanisms of green tea involving the gut-liver axis in obese models of NASH”

Frank Duna, University of Arizona – “Gut feelings about obesity and diabetes”

Ingolf Gruen, University of Missouri – “Functionality of Azadirachta Indica A. Juss”

David Dallas, Oregon State University – “Digestion of milk proteins in infants and bioactive peptide discovery”

Hord Norman, Oregon State University – “Nitrate and nitrite exposure includes differential effects on cognitive behavior and oxygen consumption during exercise in zebrafish”

Ji-Young Lee, University of Connecticut – “Novel effect of astaxanthin on the modulation of histone deacetylases and metabotypes of hepatic stellate cells for the prevention and therapy for liver fibrosis”

David Sands, Montana State University – “Global Nutrition: Why is it so difficult?”

Barry Shane, University of California, Berkeley – “Genetic influences on folate status bio-markers”

Connie Weaver, Purdue University – “Berries and bone project”

Janos Zempleni, University of Nebraska – “Delivery and alterations of microbial signals by bovine milk exosomes in non-bovine species”

Jen Teske, University of Arizona – “Sleep and obesity: using pre-clinical sleep studies to improve health”

Yanhong Liu, University of California, Davis – “Anti-inflammatory and antioxidant properties of phytochemicals”

Wendy White, Iowa State University – “Modeling the dose effects of soybean oil in salad dressing on the bioavailability of carotenoids and fat-soluble vitamins in salad vegetables”

At the end of each presentation, a short discussion took place.

The meeting adjourned on Feb. 9, 2018, at 11 AM.

Accomplishments

Members of the W3002 Multistate project have been vastly productive during the past reporting period as evidenced by the dissemination of new products, technologies, and knowledge through peer-review publications, book reviews, presentations at symposia, conferences, and in the media. Project objectives are listed below along with scholarship activities from each of the represented institutions. <ol> <li>Determine the bioavailability (absorption, distribution, metabolism, elimination) of nutrients and other food components.</li> <li>Evaluate the bioactivity of nutrients and other food components in order to elucidate their underlying protective mechanisms.</li> </ol> Kansas State University (Brian Lindshield). We collected nutrition, economic, and women's empowerment baseline information from small-holder (<5 hectares) maize Guatemalan farmers through questionnaires before distributing GrainPro Bags or plastic drums. We plan to administer the same survey again next year. We also produced some newly formulated corn-soy and sorghum soy fortified blended foods that we plan to assess the nutritional efficacy of in the near future. Outputs: we published 5 new peer reviewed journal articles since our last meeting/report (May, 2017-Feb, 2018). We hope to have all MFFAPP project manuscript published this year. Outcomes: An application to have the sorghum-cowpea fortified blended food that we developed has been submitted for inclusion on the USAID approved product list. Montana State University (David C. Sands). Background: The problem of developing food crops with higher nutritional value is that the selection of nutritional value often conflicts with selection for traits necessary for high agronomic yield. This is a problem with our field crops - peas, potatoes, wheat, oat and barley. We have developed rapid selection parameters for potatoes that are lower in glycemic index than the main commodity varieties. We now have 7 cultivars being tested for agronomic production and of these we will choose several for human glycemic index testing. One low glycemic cultivar has been selected for its higher lysine level and this is being crossed with other cultivars.  Similarly, a series of high lysine tomato cultivars have been similarly selected and these are in the F2 cross stage however it may be that they have a lysine excreting endophyte. We have developed a high throughput screening platform for the oilseed crop, Camelina sativa yielding four lines that are higher, much higher in the omega-3: omega-6 fatty acid ratio than canola, corn, soy, or safflower oils. A food processing company in Illinois is now growing this specialty crop for use of its high omega, low glucosinolate oil and it anticipates production of 40,000 acres. Proatina a naked oat is being developed for use as a high protein/low glycemic substitute for rice because of its nutritional value compared to rice (2.5x higher in protein, much lower glycemic index, absence of arsenates, and because of its low pesticide demand and dryland habit it is a more ecologically sustainable to produce. A large specialty milling company has contracted to increase production of Proatina to 20 million pounds. The market target is the global community of Type II diabetics, especially those that use rice as a food staple. Progress: This year’s research continued focus on development of new crop cultivars specifically selected for enhanced nutritional traits. This was the first year when some of our improved nutrition cultivars were produced in field scale amounts by processing and marketing concerns aimed at the nutritional food markets. These include high lysine wheats, low glycemic potatoes, a high protein-low glycemic naked oat (Proatina), and a high omega 3 selection of Camelina sativa. In each case the economic outcome and output based on the nutritional value-added aspect of the new crop had to make up for the trade-off in yield. Nutritional selection reached our milestones of commercial product interest, development and field production. The increased consumer demand for nutritionally improved crops has been prompted by basic research and physiology that defined what was needed in human nutrition and health promoting diets. The Ohio State University (Richard Bruno). Under Aim 1, the OH station has completed a clinical trial in humans that has defined the altered metabolism, pharmacokinetics, and elimination of a novel metabolite of alpha-tocopherol, i.e. α-carboxyethyl hydroxychromanol (a-CEHC), in patients with metabolic syndrome compared with healthy adults. Reduced generation and elimination of α-CEHC occurred without affecting the flux of its upstream metabolism (α-CMBHC) to α-CEHC. The findings of this investigation demonstrate that individuals with metabolic syndrome have lower α-tocopherol bioavailability, and their lower bioavailability occurs without upregulating P450-mediated metabolism of α-tocopherol to generate α-CEHC. That individuals with metabolic syndrome have substantially less generation of α-CEHC, suggests insufficient hepatic α-tocopherol status. Consequently, this milestone supports higher dietary α-tocopherol requirements for individuals with metabolic syndrome, and provides foundational basis for future efforts to validate vitamin E metabolites as novel biomarkers to assess vitamin E status. Under Aim 2, the OH station has completed preclinical studies in obese mouse models of nonalcoholic steatohepatitis (NASH) to assess the mechanism by which green tea extract (GTE) mitigates NFkB-dependent liver injury. Studies were performed in wild-type and loss-of-function TLR4-mutant mice to define the extent to which GTE protects against NASH in a TLR4-dependent manner. GTE reduces hepatic NFkB activation and histological evidence of NASH to the extent lowered in TLR4-mutant mice regardless of GTE supplementation. GTE in WT mice also decreased hepatic TLR4 and MyD88 expression. Although unaffected by genotype, GTE decreased circulating levels of endotoxin (a ligand for TLR4) and upregulated expression levels of intestinal tight junction proteins. These findings support that GTE ameliorates NASH through a mechanism involving the gut-liver axis. Specifically, GTE likely limits NFκB activation in a TLR4-dependent manner at the liver, but intestinal-level benefits that limit endotoxin translocation are apparently independent of intact TLR4 signaling. This milestone establishes evidence in preclinical models that GTE is effective in managing NASH through a mechanism involving the gut-liver axis, and our long-term goal to study GTE in humans to alleviate metabolic endotoxemia in relation to NASH risk. Oregon State University (Emily Ho). Zinc and chronic disease: Recently we have found that zinc status is compromised with age.  Zinc deficiency causes immune cells to be sensitized to inflammation and also occurs in older animals and humans.  We have also examined the impact of zinc deficiency on developmental outcomes, gut microbiota, oxidative stress and inflammation induced by arsenic. For these studies we have developed a zebrafish and rodent models to examine the interaction among zinc and arsenic.  Zinc deficiency exacerbated disturbances on the microbiome in rodents and neurobehavior in zebrafish. Plant-derived phytochemicals and cancer: We have found that sulforaphane, a chemical found in cruciferous vegetables inhibits histone deacetylases, alters DNA methylation and histone methylation, lincRNA expression and has anti-cancer properties in the prostate. We have found unique metabolomic signatures in plasma of human subjects fed broccoli sprouts.  This work may help define new biomarkers of intake and novel mechanistic targets of bioactives derived from broccoli. Outputs: 1) Identify new risk factors in prostate cancer and offer novel dietary modifications to reduce the incidence of prostate cancer; 2) Establish low cruciferous vegetable intake as a risk factor for the development of prostate cancer by altering histone modifications and cell proliferation pathways; 3) Gain knowledge of the mechanisms behind the health benefits of micronutrients and phytochemicals such as zinc and compounds derived from cruciferous vegetables; 4) Establish low dietary zinc as risk factor for inflammatory processes, DNA damage and cancer risk and identify new biomarkers for human zinc deficiency; 5) Establish function of zinc and changes in zinc metabolism with development and aging. Oregon State University (Norman Hord). Background: Dietary exposure to nitrates and nitrites is associated with cardiovascular health benefits and improved athletic performance while, in the context of processed meats consumption, increased gastrointestinal cancer risk. Humans concentrate nitrate from dietary or endogenous sources in the salivary glands, which is then reduced to nitrite, swallowed, and absorbed. Circulating nitrite acts as a reservoir for nitric oxide (NO) with its reduction to NO potentiated in acidic or hypoxic areas, such as contracting skeletal muscle. NO is an important signaling molecule with a short half-life that regulates cardiovascular function, cellular energetics, and neurotransmission. Progress: We had previously demonstrated, using two female rat models (young rats and ovariectomized rats), that dietary nitrate has no effect on bone growth, bone loss or the community structure of the gut microbiome. We extend the knowledge in this field with data described below by quantifying human exposure to dietary nitrate and nitrite and data from a zebrafish model informing how dietary nitrate may improve exercise performance by lowering the oxygen cost of physical activity (manuscript in preparation). Project 1 demonstrates the variability in the concentration of dietary nitrate in cultural food patterns and dietary supplements. Nitrate and nitrite content in foods is lacking from nutrient databases which limits the ability to study health-related epidemiological associations. Therefore, we estimated human nitrate and nitrite intakes from cultural meal patterns, foods and dietary supplements in order to determine the potential exposure range from available foods. Examination of prototypical daily meal patterns from four cultures showed meal patterns with the greatest nitrate and nitrite concentration were those with an abundant amount of leafy greens and root vegetables, such as the Japanese and Chinese diet, while concentrations in the American and Indian diet were considerably lower. Furthermore, consumption of one serving of a nitrate-rich food or supplement can exceed the World Health Organization Adequate Daily Intake (ADI) for nitrate (0-3.7 mg/kg body weight/day or 222 mg/day for a 60kg adult). The results for project 2 described below demonstrate improved muscle performance in zebrafish in response to exercise in a swim test. Inorganic nitrate improves exercise performance by reducing the oxygen cost of exercise. Nitrate is reduced into nitrite and NO in vivo, which is favored under acidic and hypoxic conditions. We hypothesized that nitrate-induced changes in mitochondrial function would stimulate changes in the utilization of metabolic fuels for energy production. Adult zebrafish were treated with sodium nitrate (606.9 mg/L) or control water for 21 days. Nitrate treatment significantly increased both blood and whole-body levels of nitrate and nitrite, demonstrating that nitrate was uptaken and reduced into nitrite. Nitrate reduced the oxygen cost of exercise during a 2-hour strenuous graded exercise test. There were no significant differences between mitochondrial function between control and nitrate-treated zebrafish, as evidenced by OROBOROS analysis of isolated mitochondria after treatment. Metabolomics analysis of whole fish illustrated that nitrate altered metabolites related to NO homeostasis, and stimulated glycolysis, ketogenesis, and fatty acid oxidation in zebrafish.    Oregon State University (David Dallas). Background: Human milk evolved to provide nourishment, immunoprotection and support for the development of a commensal-dominant gut microbiome for term infants. Milk proteins are carriers of encrypted bioactive peptides with antimicrobial, immunomodulatory, anti-hypertensive, calcium delivery and gut barrier enhancement functions. These peptides must be released during infant digestion to be biologically relevant. Preterm infants have poor protein digestion and a structurally and functionally immature digestive system. Little evolutionary adaptation to provide optimal nutrition to significantly preterm infants is likely, as these infants rarely survived prior to advances in modern medical care. Therefore, bioactive peptides that evolved to be released within the term infant may be missing in the preterm infant. Differential or sub-optimal release of these bioactive peptides in early premature infants could conceivably alter health outcomes in multiple ways. Though research evidence has shown that the premature infant gut is structurally immature and protease production is decreased, what remains unknown is how these changes impact the overall breakdown of human milk proteins and release of bioactive peptides in the premature infant. There is, therefore, a critical need to determine how prematurity affects protein digestive function, the release of bioactive peptides in the infant gut, and how that ultimately impacts the infant. Our long-term research goal is to identify the optimal protein nourishment for preterm neonates to improve developmental outcomes. Our overall objective of our several funded projects is to examine the release of peptides in the digestive tract of infants, determine their functions and determine which peptides are missing in the preterm infant gut. These projects have potential for significant impact in that the identification of the degree to which digestion and the release of bioactive peptides is impaired for preterm infants will inform ways in which mother's milk and donor milk could be augmented, for example, with enzymatic supplementation or bioactive peptide supplementation to improve infant gut health. Progress: 1) We have determined the peptides released in the stomach of term and preterm infants; 2) We have determined which milk and gastric proteases are responsible for peptide release; 3) We have identified hundreds of released peptides that are homologous with known functional peptides; 4) We have established a database of known bioactive milk peptides from across literature that is open to all. Short-term outcomes: In one year of W-3002 support, 2 postdoctoral scholars, 2 graduate students, 6 undergraduates and 1 faculty research assistant were supervised. Eight manuscripts were published and twelve research presentation were given. Our research includes collaborations with many other scientists, including neonatologists in Oregon at Oregon Health & Sciences University and Randall Children’s Hospital. Purdue University (Connie Weaver). Blueberries and other fruits with high polyphenolic content have been associated with reducing postmenopausal bone loss.  We determined the effective dose in an OVX animal model and are now conducting a clinical trial in postmenopausal women.  With collaborators, we are exploring antioxidant and anti-inflammatory mechanisms. University of Arizona (Frank Duca). Background: The role of the gut microbiota in the development of metabolic disease is becoming increasingly recognized. Recent work has shown that the small intestinal microbiota, an often overlooked microbial site, can impact intestinal sensing pathways that regulate energy and glucose homeostasis of the host. However, the mechanisms upon which alterations in small intestinal microbiota impact the gut-brain signaling axis is poorly understood. Progress: 1) We have assessed the role of prebiotics, a non-digestible carbohydrate in improving gut-brain neuronal signaling that regulates food intake and energy homeostasis, 2) We have demonstrated that prebiotics alter the small intestinal microbiota, which is associated with changes in small intestinal chemosensory machinery. Short-term outcomes: In one year of W-3002 support, 4 visiting scientists, graduate students, and undergraduate students were supervised. Three presentations were delivered and 3 abstracts were submitted to internationally recognized conferences. Our research includes collaborations with 3 other scientists both within and outside the University of Arizona University of Arizona (Jennifer Teske). The laboratory focuses on the negative impact of poor sleep and low physical activity to health. We investigate the relationship between sleep curtailment due to environmental noise and metabolism in a rodent model. We expanded these studies to include female rodents to test sex-specific effects and establish a rodent model of sleep deprivation-induced weight gain for females. These studies revealed that noise exposure reduces sleep by reducing the total time spent asleep and sleep quality indicated by an increased number of awakenings. Moreover, sleep loss due to noise exposure caused weight gain and increased feeding. Activities: Performed validation studies to determine sex-specific effects of noise exposure on sleep, wake, feeding, and weight gain in rodents. Demonstrated that the combination of consumption of a hedonic diet plus sleep disruption worsens metabolic outcomes than either alone. Milestones: To test whether noise-induced sleep loss exacerbates hedonic feeding behavior and weight gain in male and female rats. Short-term outcomes: 1) Surgery training and proficiency provided for students at all levels and international scholars; 2) Hands on anthropometrics, energy expenditure and intake training and estrous cycle determination for students at all levels and international scholars; 3) Grant writing and presentation skills training for oral and posters for students at all levels; and 4) Mentored student in proposal writing. Outputs: we have published 6 publications, got 2 grants, and presented 3 oral presentations, 4 poster presentations at different scientific conferences, and given one seminar. Long-term outcomes: Jennifer Teske received $1.3 million in funding for her research, provided research experiences for 42 individuals (4 high school students, 20 undergraduate students, 13 master’s degree student, 2 international scholars and 3 doctoral students) and provided employment for 2 employees. 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 continue to evaluate genetic risk factors for neural tube defects and to identify putative modifier genes which influence folate and vitamin B12 status, homocysteine levels, and methylation potential using a number of mouse strains and a cohort of students at Trinity College, Dublin. Vitamin B12 deficiency affects 10-15% of individuals over age 65. Circulating vitamin B12 levels can be used to diagnose deficiency but this test has significant false positive and false negative rates. We conducted genome-wide association studies (GWAS) in which we resolved total serum vitamin B12 into the fractions bound to transcobalamin and haptocorrin: two carrier proteins with very different biological properties. We replicated reported associations between circulating vitamin B12 levels and a common null variant in FUT2. This allele determines the secretor phenotype in which blood group antigens are found in non-blood body fluids. Vitamin B12 bound to haptocorrin (holoHC) remained highly associated with FUT2 rs601338 (p.Trp154Ter). Transcobalamin bound vitamin B12 (holoTC) was not influenced by this variant. HoloTC is the bioactive the form of the vitamin and is taken up by all tissues. In contrast, holoHC is only taken up by the liver. Using holoHC from individuals with known FUT2 genotypes, we demonstrated that FUT2 rs601338 genotype influences the glycosylation of haptocorrin. We then developed an experimental model demonstrating that holoHC is transported into cultured hepatic cells (HepG2) via the asialoglycoprotein receptor (ASGR). University of California, Davis (Yanhong Liu). Background: Excessive production of reactive oxygen species (ROS) renders oxidative damage to lipid membrane, protein and DNA of host cells which may lead to cell death and tissue injury. The risk of oxidative damage is high at the epithelial layer of gastrointestinal tract due to frequent exposure to environmental pathogen. Consumption of oxidized food/feed constituents (i.e. oxidized fat) and the insult from environmental stress factor (i.e. heat-stress) also heightens the production of ROS in enterocytes and compromises barrier function of gut epithelium in animals and humans. Progress: 1) We have assessed the antioxidant activities of several mint-derived essential oils with different types of in vitro models; 2) we have conducted an animal trial to evaluate the antioxidant capacities of inorganic selenium and organic selenium using pig as model; 3) we have evaluated a pig model for oxidative stress research in human. Short-term outcomes: In one year of W-3002 support, 10 visiting scientists, graduate students, undergraduate students were supervised. Two review papers and 6 meeting abstracts that were related to this research area were published. Eight presentations were delivered and 4 manuscripts were submitted to peer-reviewed journals. Our research includes collaborations with 4 other scientists. University of California, Davis (Peng Ji). Background: Iron fortification has been commonly practiced in most developed countries, even though the prevalence of iron-deficient anemia is low. A follow-up study in humans has shown that iron fortification in infants born with iron-replete status has long-term negative effect on cognitive development. We use neonatal piglets as translational model to study the impact of iron over-supplementation in early life on systemic iron homeostasis, CNS oxidative stress and social cognition. Progress: 1) We have performed two studies that evaluated different dose of iron fortification in preweanling piglets on CNS and peripheral iron accumulation, transcriptional adaptation of iron regulatory proteins, CNS lipid peroxidation, and hippocampal metabolomics; 2) We also performed sociability and radio maze behavioral test to assess impact of iron fortification on social and spatial cognition. 3) We conduct in vitro cell culture work to study the regulatory mechanism of cellular iron homeostasis. The results of our studies suggested iron fortification significantly increase CNS iron deposition that is associated with increased lipid peroxidation. Similarly, hippocampal metaoblomics results highlighted pathways associated with purine metabolism were interrupted. Genes encoding iron regulatory proteins demonstrate a coordinated change in the manner to inhibit further iron uptake. Piglets that received high iron supplementation had suboptimal sociability in comparison with non-supplemented control group. The results are novel, and we are working on manuscripts of the study. The cell culture work is still in pilot tests to optimize culture, treatment, and testing conditions. Some preliminary results have validate that iron treatment significantly increased intracellular iron content in gut epithelial cells. University of Connecticut (Ji-Young Lee; Sung Koo). Endotoxin tolerance is a phenomenon where exposure of innate immune cells to lipopolysaccharide (LPS) induces a refractory state to subsequent endotoxin exposures. The goal of this study was to investigate if Spirulina platensis extract (SPE) induces an endotoxin tolerance-like state. We used splenocytes and peritoneal macrophages from C57BL/6J mice fed a high fat/high sucrose (HF/HS) control or a HF/HS diet containing 0.25% (w/w) SPE for 16 weeks for ex vivo LPS stimulation and endotoxin tolerant macrophages to evaluate the effects of SPE on endotoxin tolerance. Cells from SPE-fed mice displayed significantly less expression of pro-inflammatory genes than those from control mice. Endotoxin tolerant (ET) macrophages were produced in vitro by incubating RAW 264.7 macrophages with low-dose LPS to determine the energy phenotype of naïve, SPE-treated, and ET macrophages. Compared to naïve macrophages exposed to a high-dose LPS (100 ng/mL) for the first time, ET macrophages showed significantly less pro-inflammatory gene expression after LPS stimulation, which was also observed with SPE treatment. Consistently, nuclear translocation of p65 was markedly reduced in both ET and SPE-treated macrophages upon LPS stimulation with increase in nuclear protein levels of p50 and B-cell lymphoma 3- encoded protein. In conclusion, the anti-inflammatory effect of SPE is at least partly attributable to the induction of an endotoxin tolerance-like state in macrophages, which shares common characteristics of macrophage endotoxin tolerance. Activation of hepatic stellate cells (HSCs) is critical for liver fibrosis development. Previously, we showed that astaxanthin (ASTX), a xanthophyll carotenoid, has anti-fibrogenic effects in LX-2 cells, a human HSC cell line. We sought to determine the effect of ASTX on HSC activation, and to identify molecular mediators that are critically involved in the processes. ASTX prevented the activation of mouse primary HSCs, as evidenced by attenuated induction of procollagen type I a1. In human primary HSCs, ASTX also inhibited transforming growth factor b1 (TGFβ1)-induced fibrogenic gene expression. Among 11 classical histone deacetylases (HDACs), difference in HDAC9 mRNA levels between quiescent and activated HSCs was most evident while ASTX significantly decreased the expression of HDAC9 and its transcriptional regulator myocyte enhancer factor 2 (MEF2). ASTX decreased HDAC9 protein as well. In the activated HSCs, ASTX significantly reduced mRNA of HDAC9 and MEF2. Human primary biliary cirrhosis livers showed significantly higher HDAC9 mRNA and protein levels than normal livers, and other liver pathologies also exhibited induced HDAC9 expression. HDAC9 knockdown in LX-2 cells decreased TGFβ1-induced fibrogenic gene expression. In conclusion, ASTX inhibits HSC activation and facilitates HSC inactivation, which is attributable to its inhibitory action on HDAC9 expression. We previously showed that polyphenol-rich blackcurrant extract (BCE) showed a hypocholeseterolemic effect in mice fed a high fat diet. As direct cholesterol removal from the body via the intestine has been recently appreciated, we investigated the effect of BCE on the modulation of genes involved in intestinal cholesterol transport using Caco-2 cells as an in vitro model. BCE significantly increased protein levels of low-density lipoprotein receptor (LDLR) without altering its mRNA, which consequently increased low-density lipoprotein uptake into Caco-2 cells. This post-transcriptional induction of LDLR by BCE was markedly attenuated in the presence of rapamycin, an inhibitor of mechanistic target of rapamycin complex 1 (mTORC1). In addition, BCE altered genes involved in cholesterol transport in the enterocytes, including apical and basolateral cholesterol transporters, in such a way that could enhance cholesterol flux from the basolateral to apical side of the enterocytes. Indeed, BCE significantly increased the flux of LDL-derived cholesterol from the basolateral to the apical chamber of Caco-2 monolayer. LDLR protein levels were markedly increased by anthocyanin-rich fraction, but not by anthocyanin-free fraction. In conclusion, the mTORC1-dependent post-transcriptional induction of LDLR by BCE anthocyanins drove the transport of LDL- derived cholesterol to the apical side of the enterocytes. This may represent a potential mechanism for the hypocholesterolemic effect of BCE. We previously showed that the organic extract of a blue-green alga, Spirulina platensis (SPE) had potent anti-inflammatory effects in macrophages. We investigated the contribution of the anti-inflammatory effects of SPE in macrophages to adipogenesis/lipogenesis in 3T3-L1 adipocytes. 3T3-L1 preadipocytes were treated with 10% conditioned medium from LPS-stimulated RAW 264.7 macrophages (CMC) or LPS-stimulated but SPE-pretreated macrophages (CMS) at different stages of adipocyte differentiation. The expression of adipocyte differentiation markers were significantly repressed by CMC, while the repression was attenuated by CMS. Oil Red O staining confirmed that adipocyte maturation in CMS-treated cells, but not in CMC-treated cells, was equivalent to that of control cells. In lipid-laden adipocytes, CMC promoted the loss of lipid droplets while CMS had minimal effects. Histone deacetylase 9 mRNA and protein levels were increased during adipocyte maturation, which were decreased by CMC. In conclusion, by cross-talking with adipocytes, the anti-inflammatory effects of SPE in macrophages promoted adipocyte differentiation/maturation, at least in part, by repressing the activation of NF-kB inflammatory pathways, which otherwise can be compromised in inflammatory conditions. Endotoxin tolerance is a phenomenon where exposure of innate immune cells to lipopolysaccharide (LPS) induces a refractory state to subsequent endotoxin exposures, resulting in a lack of pro-inflammatory cytokine and chemokine production. We evaluated whether SPE induces endotoxin tolerance-like state to exert its anti-inflammatory effect and to determine its effect of macrophage energy phenotype. Splenocytes and resident peritoneal macrophages from mice fed a HF/high sucrose (HF/HS) diet containing SPE displayed significantly less expression of pro-inflammatory genes than those from control mice. We produced endotoxin tolerant (ET) macrophages in vitro by incubating RAW 264.7 macrophages with low-dose of LPS. Compared to naïve macrophages exposed to a high-dose LPS for the first time, ET macrophages showed significantly less pro-inflammatory gene expression after LPS stimulation, which was also observed with SPE treatment. Consistently, nuclear translocation of p65 was markedly reduced in both ET and SPE-treated macrophages upon LPS stimulation. Both SPE-treated and ET macrophages had increased mRNA and nuclear protein levels of p50 and B-cell lymphoma 3-encoded protein, which play a critical role in endotoxin tolerance. Both SPE-treated and ET macrophages had similar energy phenotype in that they relied more on glycolysis than mitochondrial respiration for energy generation compared to naïve cells. In conclusion, the anti-inflammatory effect of SPE is at least partly attributable to the induction of an endotoxin tolerance-like state in macrophages, which shares common characteristics of macrophage endotoxin tolerance. University of Illinois, Urbana-Champaign (Juan Andrade). Background: Our research focuses on developing technologies or approaches to improve nutrition in vulnerable populations in the US and abroad. Efforts have been focused on: 1) characterization of legume-based protein nanoaggregates to improve stability and delivery of fat-soluble bioactives and nutrients; and, 2) characterization of lipid-based nutrition supplement (LNS) to prevent and control severe acute malnutrition (SAM) and parasitic infection. Outputs: 1) Characterization of legume-based protein nanoparticles. A legume-based nanoparticle system can be advantageous for the delivery of fat soluble compounds that can address nutrition gaps due to its enhanced stability and bioavailability. Nanoaggregates containing vitamin D were characterized in terms of stability and bioavailability in animal models. We found that dispersion of vitamin D in nanoaggregates enhances its oral bioavailability as evaluated in rats. 2) Lipid-based nutrition supplement (LNS) characterization. We evaluated the addition of essential oils into an LNS designed for the Indian context. We found that oregano essential oil bioactives were capable of reducing infection of Cryptosporidium using an absorptive cell model. We also found that encapsulation of bioactives into cyclodextrin particles allows for the feasible, target delivery of bioactives as encapsulation enhances delivery, resist digestion conditions, and do not affect sensory attributes of LNS. University of Maine (Dorothy Klimis-Zacas). Atherosclerosis is a chronic inflammatory, progressive disease of the large arteries that can lead to CVD and stroke. Angiogenesis is the formation of new capillary blood vessels from existing ones and endothelial cell migration and proliferation contribute to the development of angiogenesis; critical in the early stages of atherosclerosis. Wild blueberries (Vaccinium angustifolium) are rich in anthocyanins (ACNs) and phenolic acids (Phen) having an exceptional ranking for antioxidant capacity compared to other berries and fruits. During 2017-2018 we continued to investigate the effect of ACNs and Phen fractions and their combination on endothelial cell migration and angiogenesis and mechanisms thereof, by exploring relevant biomarkers of cell migration and angiogenesis such as RhoA, Rac1 GTPases, AKT, VEGF and eNOS and their gene expression, critical for cell morphology, cytoskeleton integrity, cell permeability, angiogenesis and cell migration. Preliminary results document a differential response of the above bioactive compounds on endothelial cell migration and relevant proteins and their gene expression, based on type of fraction and its concentration. Studies on angiogenesis also document a concentration-depended effect, critical concentrations at which angiogenesis is modulated and differential response based on the different fractions (ACNs or Phen). ACNs seem to inhibit HUVEC migration and angiogenesis while PAs promote this process. Another project initiated in 2017, targets the role of Red Raspberries (Rubus idaeus) on endothelial function (vasoconstriction and vasodilation) and obesity-induced inflammation by assessing pro-inflammatory markers and their gene expression in hepatic and adipose tissues in an animal model of the Metabolic Syndrome (MetS), the Obese Zucker Rat (OZR). Preliminary results support the role of Red Raspberries in normalizing the endothelial dysfunction and attenuating inflammation associated with MetS. Outputs: Results from the period covered in this report were disseminated in form of a poster at the International Conference of Polyphenols and Health in Quebec, Canada (October 2017), at the WBANA Health Summit in Bar Harbor, Maine (September, 2017) in form of an oral presentation, at the Berry Health Benefits Symposium in Pismo Bay, California (March, 2017) in form of a poster, which received second price in the poster competition, and results will be presented at the American Society for Nutrition Meetings, in Boston, MA (April, 2018). Additionally, several manuscripts are in preparation to be submitted by Summer 2018. Other outputs will be presentations at invited lectures and through the media (newspaper articles, blogs and interviews). A provisional application to EFS for a patent is pending to commercialize the findings related to the role of phenolics on cell migration and angiogenesis. University of Massachusetts (Yeonhwa Park; Hang Xiao). We have studied various bioactive food components to improve bioaccesibility and bioavailability and determined the bioactivites of food bioactivities using various models. We have tested compounds, such as piceatannol, 3,3'-Diindolylmethane, β-Carotene, and nobiletin during this period. University of Missouri (Ingolf Gruen). Neem (Azadirachta indica) is an evergreen tree cultivated in various parts of the Indian sub-continent. It has been in use over centuries in the Indian folk medicine for its therapeutic value. Given the prominent role it has played in curing diseases of the villagers over centuries, it has been hailed as a “divine” tree, a “village dispensary” and “ nature’s drugstore” .Today, extensive research has shown that it may have anti-cancer, anti- diabetic, anti-inflammatory, anti-ulcerogenic, and anti-microbial effects. However, there are significant gaps in the scientific literature about the compounds contributing to its medicinal potential. In this study, select flavonoids, namely flavonols: myricetin, quercetin and kaempferol are quantified by a HPLC method in Neem powder and leaves as well as green and black tea leaves for comparison. It was observed that Neem dry powder had significantly higher (p< 0.05) flavonol content at 11.146 ±1.09 mg/g than green and black tea 5.782 ± 0.723 mg/g and 5.599 ± 0.484 mg/g respectively. The tea infusions of the plant materials also yielded similar results. The total phenolics were found to be the highest in green tea infusion because of its high flavan-3-ol content, followed by black tea, Neem powder and Neem tea cut leaves with values of 174.55 ± 12.85, 106.85 ± 6.511, 55.60 ± 7.90 and 87. 35 ± 7.42 mg/g respectively. DPPH and FRAP anti-oxidant assays yielded contradictory results which can be explained by the very principle of these assays. It was also observed that the particle size of ground tea leaves significantly influenced the yield of flavonols, phenolics and anti-oxidant assay. In addition, neem is extremely bitter, in large part due to its limonoid content, making it unpalatable and limiting its potential use in dietary supplements or foods alike. Bitterness reduction, especially of foods and beverages containing phytonutrients, is one of the biggest challenges in the food industry. The objective of this experiment of our overall study was to apply two adsorbent based strategies, namely solid phase extraction (SPE) and Amberlite XAD-16 (AMB) resin, to achieve de-bittering of neem tea and to determine the effects of the de-bittering on the bio-active, color, and volatile properties. The solid SPE treatment completely removed the flavonol, quercetin, from neem tea while in Amberlite XAD-16 treated tea (AMB) it was only insignificantly (p > 0.05) reduced. We also observed decreases in total phenolic content and consequently anti-oxidant activities after de-bittering. A 62% mean reduction of limonoid aglycones indicated diminished levels of bitterness. The loss of phenolics lead to a visually appreciable color change in the treated teas. The de-bittering also leads to a loss of sesquiterpenes, ketones and acids from neem tea. In conclusion, we found that while SPE cartridges were more efficient in removing bitterness, they caused a greater reduction in bio-active compounds than AMB XAD-16 resins, which may ultimately affect the health properties of neem tea. Over the last year, in the context of this project, we initiated an investigation into the bioactive compounds and the resulting bitterness of cacao and chocolate, specifically our goal is to identify and quantify the complex mixture of important bitter compounds within the three compound classes that are known to be important to cocoa--methylxanthines, flavan-3-ols, and diketopiperazines. Chocolate is made from the fermented, dried, and roasted seeds of the Theobroma cacao tree. It is now understood that cacao, also known in English-speaking countries as cocoa, contains a variety of compounds, including well-known flavonoid polyphenols, the consumption of which has positive impacts on human heart health and blood pressure, cancer reduction, LDL cholesterol reduction, and insulin resistance improvements, as numerous in vitro, in vivo, and observational studies have confirmed. In important research published in 2017, performed, in part, by Harvard's Chan School of Public Health, a participant observational study, composed of men and women between the ages of 50 and 64, led to the determination that chocolate consumption is inversely correlated with atrial fibrillation (AR). This is an important finding, as AR is associated with multiple significant health problems such as "higher risk of stroke, heart failure, cognitive decline, dementia and mortality." A persuasive case has, therefore, been made that cacao is a healthful addition to a balanced diet. However, despite the strong evidence for the healthful qualities of cacao, which has led to an overall increase in consumer purchases of more high-cacao-content chocolate than in previous decades, sales figures still show that there is an unwillingness by many Americans to consume higher-cacao-content chocolate, which tends to be more bitter, a taste modality not readily appreciated by most. Therefore, if the bitterness of cacao could be minimized, higher-cacao-content and lower-sugar chocolate confection sales could capture an even larger segment of the conventional and healthy snack-food market by achieving the elusive combination of being both tasty and healthy. The key is to better grasp the causes of bitterness in cacao, which are complex and still not well understood. What has long been known is that the methylxanthine, theobromine, is an important contributing bitter compound in cacao, with its name even being derived from the genus of the cacao tree, Theobroma. Similarly, the related methylxanthine, caffeine, which is well known as an important bitter compound in coffee, has also been found to impart bitterness in cacao and chocolate. Certain flavan-3-ols, which are a subclass of the aforementioned healthy flavonoids, and which are found in black tea, are also bitter and present in cacao at levels above the detection threshold, especially the compound (-)-epicatechin. After project initiation, cocoa beans were obtained from Ghana, Madagascar, and Peru and have been roasted according to a standard roasting profile by chocolate manufacturer Patric Chocolate, LLC. Methods have been developed and validated, and standard curves have been established for the cocoa bitter compounds: Caffeine, Theobromine, Catechin, Epicatechin, and selected Diketopiperazines (DKPs), including cyclo(L-Pro-L-Val) which according to the most recent literature available is considered the most important DKP in regards to cocoa bitterness. The bitter compounds caffeine, Theobromine, Catechin, and Epicatechin were found in all samples. While a preliminary analysis of the data showed, as expected, some variation, we still need to analyze the data statistically. We were not able to replicate the finding reported by Stark (Stark T, Bareuther S, Hofmann T. Molecular definition of the taste of roasted cocoa nibs (Theobroma cacao) by means of quantitative studies and sensory experiments. J Agr Food Chem. 2006;54(15):5530-9) that the diketopiperazine cyclo(L-Pro-L-Val) is an important bitter compound! While we were able to positively identify cyclo(L-Pro-L-Val) in a very dark roasted cocoa using LC-MS-MS, which then allowed us to also detect and quantify this diketopiperazine in other roasts, the concentration of cyclo(L-Pro-L-Val) is apparently considerably lower, i.e. only about 5-10% of that reported by Stark, which put its concentration at or below its sensory threshold. However, since Stark's sample was not well characterized, i.e. no information was given on roast profile or how many samples were analyzed, a direct comparison of results is difficult. On the other hand, we were able to detect this diketopiperazine even in unroasted samples, indicating that DKPs are formed already during the fermentation stage of the beans. Neem (Azadirachta indica) is an evergreen tree cultivated in various parts of the Indian sub-continent. It has been in use over centuries in the Indian folk medicine for its therapeutic value. Extensive research has shown that it may have anti-cancer, anti-diabetic, anti-inflammatory, anti-ulcerogenic, and anti-microbial effects. However, neem is extremely bitter, in large part due to its limonoid content, making it unpalatable and limiting its potential use in dietary supplements or foods alike. The objective of this experiment was to apply two adsorbent based strategies, namely solid phase extraction and Amberlite XAD-16 (AMB) resin, to achieve de-bittering of neem tea and to determine the effects of the de-bittering on the bio-active, color, and volatile properties. The solid phase extraction treatment completely removed the flavonol, quercetin, from neem tea while in Amberlite XAD-16 treated tea (AMB) it was only insignificantly reduced. We also observed decreases in total phenolic content and consequently anti-oxidant activities after de-bittering. A 62% mean reduction of limonoid aglycones indicated diminished levels of bitterness. The loss of phenolics lead to a visually appreciable color change in the treated teas. The de-bittering also leads to a loss of sesquiterpenes, ketones and acids from neem tea. In conclusion, we found that while solid phase extraction cartridges were more efficient in removing bitterness, they caused a greater reduction in bio-active compounds than AMB XAD-16 resins, which may ultimately affect the health properties of neem tea. University of Nebraska, Lincoln (Janos Zempleni). Background: Virtually every cells produces and secretes exosomes (nanoparticles) loaded with cargos such as various species of RNAs, proteins and lipids. Exosomes play essential roles in cell-to-cell communication. The transfer of exosomes cargos from donor cells to receptor cells alters gene expression and metabolism in receptor cells. We have made the paradigm-shifting discovery that exosomes and their cargos are not only obtained through endogenous synthesis but also from dietary sources such as bovine milk and chicken eggs. Progress: 1) We have assessed the microRNA cargos in chicken egg yolk, the bioavailability of chicken egg exosomes and their microRNA cargos in humans, and the effects of exosomes and cargos in peripheral blood mononuclear cells. 2) We have conducted a comprehensive analysis of the bioavailability and distribution of fluorophore-labeled milk exosomes and microRNAs in mice. 3) We have developed an exosome and cargo tracking (ECT) mouse. 4) We have assessed storage stability and microRNA cargos  in human milk, bovine milk and infant formulas by RNA-sequencing analysis and qRT-PCR. 5) We have assessed the role of glycoproteins on the surface of milk exosomes in absorption and distribution of bovine milk exosomes in mice. 6) We have characterized the composition of exosome-defined, AIN-93G-based diets developed in our laboratory. 7) We demonstrated that depletion of milk exosomes and their RNA cargos elicits phenotypes such as impaired fecundity, aberrant purine metabolism, altered immune function, impaired spatial learning and memory, and changes in the gut microbiome. 8) We have generated preliminary data suggesting that bovine milk exosomes deliver a large load of microbial RNAs to hosts. Short-term outcomes: In five years of W-3002 support, 21 visiting scientists, postdoctoral fellows, graduate students, undergraduate students, and staff were supervised. Fourteen papers, six reviews, one book chapter, 15 meeting abstracts and five outreach publications (newspapers, magazines) were published, and 33 presentations were delivered. A public website was created that disseminates information about dietary microRNAs (Shu et al.). Our research includes collaborations with one scientist from this W-3002 group and 13 other scientists. We have filed a provisional patent application Extracellular Vesicles and Methods of Using, on 3/15/2017 (62/471,572). Iowa State University (Wendy White). Background: Based on the bioavailability of carotenoids and fat-soluble vitamins measured in our human study, we developed statistical models to predict the relation between the amount of soybean oil in salad dressing and the absorption of: 1) alpha-carotene, beta-carotene, and lycopene in salad vegetables; and 2) retinyl palmitate, the major form of vitamin A absorbed after the intestinal metabolism of alpha- and beta-carotene. For all carotenoids and fat-soluble vitamins studied, we provided important new information regarding the optimal amount of vegetable oil needed to promote their absorption from fresh vegetables. We evaluated the effects of increasing amounts of vegetable oil in salad dressing on the absorption and bioactivity of alphacarotene and beta-carotene. Bioactivity was measured as the retinyl palmitate (vitamin A) formed through the intestinal metabolism of these nutrients. This project has provided extensive training for the Principal Investigator in the form of four summers of self-study in the use of statistical models to predict the intestinal absorption of nutrients. Outcomes: A manuscript describing the results was published in the American Journal of Clinical Nutrition, the top-ranked nutrition journal worldwide. The manuscript was selected to be the focus of an editorial published in the same issue of that journal. The study results were disseminated via a university press release, which was then covered by national and international media outlets, including LinkedIn, the Daily Mail (United Kingdom), Men's Health, Men's Fitness, AARP Magazine, New Indian Express (India), KPCC Radio (Southern California), NutraIngredients, and Science Daily. The study findings were presented at the annual meeting of the American Society for Nutrition in conjunction with the Experimental Biology '17 conference in Chicago, IL.

Publications

Abelilla, J. J., Y. Liu, and H. H. Stein. 2018. Digestible indispensable amino acid score (DIAAS) and protein digestibility corrected amino acid score (PDCAAS) in oat protein concentrate measured in 20- to 30-kilogram pigs. J. Sci. Food Agri. 98:410-414. Aguilar-Lozano A, Baier SR, Grove R, Shu J, Giraud D, Mercer KE, Cui J, Badger TM, Adamec J, Andres A, Zempleni J. Concentrations of purine metabolites are elevated in human fluids from adults and infants and in livers from mice fed diets depleted of bovine milk exosomes and their RNA cargos (submitted). Ahluwalia, A., Gladwin, M., Coleman, G.D., Hord, N. G., Howard, G., Kim-Shapiro, D., Lajous, M., Larsen, F., Lefer, D.J., McClure, L.A., Nolan, B.T., Pluta, R., Schechter, A., Wang, C.-Y., Ward, M.H., and Harman, J.L. (2016) Dietary nitrate and the epidemiology of cardiovascular disease: Report from a National Heart, Lung, and Blood Institute Workshop, Journal of the American Heart Association J Am Heart Assoc. 2016 Jul 6;5(7). doi: 10.1161/JAHA.116.003402. Bae M., Y. Park, J. Lee. Comprehensive review on food components with anti-fibrotic activity for the prevention of liver fibrosis. J Nutr Biochem 2018; 55:1-11. Epub ahead of print on November 16, 2017. Bai, L. L., F. Wu, H. Liu, S. Zhang, L. Liu, X. S. Piao, Y. H. Liu, P. A. Thacker, and F. L. Wang. 2017. Effects of dietary calcium levels on growth performance and bone characteristics in pigs in grower-finisher-translational phase. Anim. Feed Sci. Technol. 224:59-65. Bai, M., H. Liu, K. Xu, B. Zou, R. Yu, Y. Liu, W. Xing, H. Du, Y. Li, and Y. Yin. 2017. Effects of dietary coated cysteamine hydrochloride on pork color in finishing pigs. J. Sci. Food Agric. doi: 10.1002/jsfa.8647. 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Impact Statements

Given the low vegetable consumption by Americans, there is a need to better define the role of vegetable oil as a factor in the absorption of carotenoids and fat-soluble vitamins from these food sources. We developed statistical models to predict the effects of given amounts of soybean oil in salad dressing upon the absorption of: 1) alphacarotene, beta-carotene,and lycopene in salad vegetables; and 2) retinyl palmitate, the major form of vitamin A absorbed after the intestinal metabolism of alpha- and beta-carotene. Overall, our data indicate that even small amounts of vegetable oil significantly enhance the absorption of carotenoids, phylloquinone, and tocopherols from fresh vegetables. These data will enable nutritional scientists to develop dietary guidelines for the public regarding the amount of vegetable oil necessary to be consumed with fresh vegetables in order to maximize their health benefits and prevent chronic diseases. Iowa State University (Wendy White).