Brief Summary of Minutes of Annual Meeting

The participants of this multi-state project have been highly productive during the past reporting period as evidenced by >50 peer-reviewed publications among attending participants and enhancement of collaborations between project members. Project objectives are listed below along with scholarly activities of the lead station. Objective 1): Determine the bioavailability (absorption, distribution, metabolism, elimination) of nutrients and other food components and their environmental and genetic determinants. Objective 2): Evaluate the bioactivity of nutrients and other food components in order to elucidate their underlying protective mechanisms. Colorado State University (Jairam Vanamala). We developed a systematic approach to screen storage and processing effects on bioactivity of plant products (colored potatoes) by focusing on (i) analysis of both storage and processing (baking and chipping) on phenolic content (TP), composition, sensory attributes and biological properties (anti-proliferation and pro-apoptosis); and (ii) investigation of consumer and producer preferences for specialty potato products. We have also collected extensive samples from pigs (n = 40) consuming a high-fat diet for 12 wk followed by supplementation with white vs. purple potatoes at two different doses (10% vs. 20% of diet) with an aim to determine the appropriate dose needed to suppress an inflammatory cascade. We demonstrated that post-harvest storage of colored potatoes elevated TP and anti-oxidant activity (AA), but suppressed anti-proliferative and pro-apoptotic properties against human colon cancer cell lines. Storage also altered the LC-MS metabolite profile of all potato varieties tested. These results suggest that it is critical to use both analytical and biological assays in conjunction to determine the post-harvest storage and processing effects on bioactivity of plant products. Chipping, but not baking, altered the metabolite profile. Chips had approximately 5-10 times less bioactivity compared to the baked or fresh potatoes. To study the effect of storage and inter-varietal differences on chipped and baked potato sensory attributes, 114 untrained panelists rated the 7 potato cultivars using a 9-point hedonic scale at 30 and 90 days of storage. In general, phenolic content was inversely related to sensory attributes. These results suggest that it is critical to consider sensory attributes along with bioactive compound profile in developing health food products. Consumers were also more willing to pay higher premium for colored potato products if they are educated on health benefits. Kansas State University (Denis Medeiros). We have ongoing work examining the impact of copper-deficiency on cardiac chaperone proteins. We have for years studied the mechanisms as to why copper deficiency leads to cardiac hypertrophy and cardiomyopathy. We have described pathology of mitochondria including the gene program responsible for mitochondrial biogenesis. During the past reporting period, we detailed the impact copper deficiency on cardiac chaperone proteins, which are integral for ferrying copper between cell compartments in which copper is used, such as the synthesis of copper containing enzymes. The copper chaperone proteins we evaluated as a result of copper deficiency include CCS, Sco1, Cox 17, and Ctr-1. Rats were fed diets either adequate or deficient in copper from weanling for 5 wk thereafter. Copper deficiency did not change Ctr-1 or Cox 17 expression. Ctr-1 is thought to regulate the amount of copper entering the myocyte or heart cell. We had expected that this protein would increase, but it did not. Cox 17 is involved in brining copper to the mitochondria. Sco1 and CCS were two chaperone proteins that increased in the heart as a result of copper deficiency. Sco1 delivers the copper to cytochrome C oxidase and CCS to the site of Cu, Zn superoxide dismutase synthesis. We also studied the impact of copper deficiency on cytochrome C oxidase subunits I and IV. We show that nuclear encoded subunits of cytochrome c oxidase were down regulated. In our earlier studies, we used a polyclonal antibody to formulate to this conclusion. We re-performed our study using mono-clonal antibodies, as other have used to determine if the source of antibodies could explain this discrepancy. Using mono-clonal antibodies we found that mitochondria encoded subunit I (Cox I) and the nuclear encoded subunit IV (Cox IV) were both significantly reduced, in contrast to what we previously reported. This suggests that copper deficiency may be causing a change in the mitochondrial encoded subunits of cytochrome c oxidase whereby the monoclonal antibodies are perhaps able to bind to the mature form of the protein and the polyclonal form may be binding to the precursor or immature form of the protein. During the past reporting period, our laboratory has also successfully completed an exhaustive review regarding the role of iron in bone metabolism. The information is now being utilized to develop a strategy on how to better translate the findings of animal models on a high risk human population and to determine if iron status is a predictor of lower bone mineral density, and stress fractures in particular, in humans. Michigan State University (Norm Hord). Dietary exposure to nitrates and nitrites is associated with cardiovascular health benefits and, in the context of processed meats consumption, gastrointestinal cancer risk. Our work describes the scientific basis for these risk assessments by examining the effect of nitrite on phenotypes associated with colorectal cancer (CRC) risk in human carcinoma cells. Nitrite and nitrate used exogenously in processed meats to enhance taste and microbial food safety is associated with increased cancer risk, but these compounds are also found in vegetables, and associated with lower cancer risk. To clarify this controversy, we determined whether nitrite promotes or blocks phenotypes associated with the promotion of cancer in colon epithelial cells. The effect of nitrite on proliferation and invasion of 4 different stages of colorectal epithelial cancer was examined. Nitrite has no effect on stage 1 SW1116 colon cancer cell at any concentrations; nitrite inhibits stage 2 SW480 proliferation at 10nm-100µM and inhibits stage 3 HCT15 at 100nm and 1µM, but promotes proliferation in stage 4 cells at 100µM. Nitrite also inhibits stage 3 SW480 cells invasion in a dose-dependent manner. However, it significantly promotes the invasion of stage 4 cells at the highest concentration. FACS data indicates that nitrite dose-dependently decreased cell cycle progression in SW480 and HCT15 with changed G2/M transition and delayed G1 phase entry. However, 100µM nitrite promoted cell cycle progression in COLO205 cells with increased S-phase entry. Taken together, nitrite can inhibit cancer cell progression at low doses and early stage but may promote cancer cell progression at higher doses in stage 4 colon cancer indicating a dose and stage dependent effect. Subsequent studies were performed to estimate exposure to nitrates and nitrites from human, bovine and formula milks relative to regulatory intake limits. The World Health Organization and American Academy of Pediatrics recommends exclusive consumption of human milk for the first 6 mo of life because of the nutritional and immunological benefits for the infant. Consumption of formula, bovine, and soy milk may be used as alternatives to human milk for infants. Human milk concentrations of colostrum (expressed d 13 postpartum; n1/412), transition milk (expressed d 37 postpartum; n1/417), and mature milk (expressed >7 d postpartum; n1/450) were 0.08 mg/100mL nitrite and 0.19 mg/100mL nitrate, 0.001mg/100mL nitrite and 0.52mg/100mL nitrate, and 0.001mg/100mL nitrite and 0.3 mg/100mL nitrate, respectively, revealing that the concentrations of these anions change as the composition of milk changes. When expressed as a percentage of the World Health Organizations Acceptable Daily Intake limits, Silk® Soy Vanilla (WhiteWave Foods, Broomfield, CO) intake could result in high nitrate intakes (104% of this standard), while intake of Bright Beginnings Soy Pediatric® formula (PBM Nutritionals, Georgia, VT) could result in the highest nitrite intakes (383% of this standard). The temporal relationship between the provision of nitrite in human milk and the development of commensal microbiota capable of reducing dietary nitrate to nitrite supports a hypothesis that humans are adapted to provide nitrite to the gastrointestinal tract from birth. These data also support the hypothesis that the high concentrations of breast milk nitrite and nitrate are evidence for a physiologic requirement to support gastrointestinal and immune homeostasis in the neonate. Montana State University (David Sands). High intake of starchy (high glycemic) grains, legumes and potatoes is one of the leading causes of obesity and heart disease. In response, MSU has recently released a new variety of pea that has very low branched starch and can be used for production of low glycemic index flour and food products. Additional low glycemic varieties of legumes can be developed. We are actively expanding our selection of low glycemic varieties to include spring, winter and durum wheats, legumes, and potatoes. Such low glycemic index crops could demand a premium in price at the farm gate. Similarly, we will continue to develop high omega-3 and high vitamin E camelina for both food and animal feed, because of its benefits to humans and animals. MSU biochemists and geneticists will follow nutritionally important genes through crosses, using recently developed and sophisticated molecular (but not transgenic) techniques, greatly reducing the time to improve crops. The resultant varieties will be tested for growth and productivity statewide at the MSU Agriculture Research Centers and the nutritional composition of the new varieties will be assessed. New research collaborations have also been identified with group members (Bruno/UConn; Vanamala/Colorado) to examine the potential bioactivity of specialty crops in physiologically relevant model systems. Ohio State University (Mark Failla) has been studying the impact of type of dietary fat on transport of carotenoids and vitamin E across the Caco-2 human intestinal cell line. B-carotene, lutein and ±-tocopherol transport into the basolateral compartment was increased to a greater extent when cells were exposed to micelles containing free fatty acids mimicking the fatty acyl composition of olive, canola and soybean oils compared to micelles with fatty acyl profile mimicking butter. The amounts of ²-carotene, lutein and ±-tocopherol transported into the basolateral compartment was highly correlated with the amounts of triacylglycerides and apoB secreted. The carotenoids were primarily present in chylomicrons and vitamin E was distributed in both chylomicron and VLDL fractions. These data suggest that dietary triacylglycerides rich in unsaturated fatty acids and particularly oleate promote the absorption of carotenoids and vitamin E by stimulating the assembly and secretion of chylomicrons. The bioaccessibility and absorption of the 24-carbon carotenoid norbixin was also investigated using the coupled in vitro digestion/Caco-2 cell model. Norbixin, an abundant carotenoid in annatto, is extensively used as a natural coloring agent by the food industry. Norbixin added to milk was highly stable during simulated digestion and bile salts enhanced partitioning into the bioaccessible fraction, suggesting delivery to the apical membrane of absorptive cells in micelles. Uptake of norbixin was proportional to apical content and both all trans and cis isomers were transported across the monolayer. Oklahoma State University. Work from Dr. Lucas demonstrates that mango exerts important bioactivities that may mitigate the risk of obesity. Mango (Mangifera indica L.) provides a number of bioactive compounds that may play a role in the attenuation of excess body fat. The effects of freeze-dried mango fruit on body composition were examined in two separate studies involving mice fed high fat diet and ovariectomized (ovx) mice. In the first study, male C57BL/6J mice were randomly divided into four treatment groups (n=8/group): control, high fat diet (HF), HF + 1% or 10% mango (w/w). After 8 wk of treatment, mice receiving the HF diet had significantly higher epididymal fat mass and % body fat. Both doses of mango prevented the increase in epididymal fat mass and % body fat due to HF diet. In the second study, female C57BL/6 mice sham-operated and ovx and were randomly assigned to receive control diet, 5% or 25% (w/w) mango diet (n=8/group). After 8 wk of treatment, ovx mice had higher % body fat in comparison to the sham group. Both doses of mango had lower % body fat in comparison to the ovx group but not quite to the level of the sham group. The mechanism by which mango exerts this positive effects on body composition is being actively investigated. Oregon State (Emily Ho) has been actively examining the role of epigenetic alterations in the risk for cancer and the extent to which bioactive nutrients such as sulforaphane, catechins and zinc regulate the mechanisms related to cancer and chronic disease development. In both animal models and humans, dietary zinc deficiency increases DNA damage in peripheral blood cells. Importantly, these functional changes precede any changes in plasma zinc levels. Zinc status is also compromised with age with ongoing studies demonstrating that zinc supplementation in older animals reverses age-related zinc deficiency and inhibits age-related immune defects and inflammatory processes. Preliminary evidence further suggests that zinc alters DNA methylation patterns and may be a novel mechanism by which zinc affects gene expression. Separate studies examining plant-derived phytochemicals from tea and cruciferous vegetables have provide evidence of the chemoprotective effects of diet. Indeed, studies show that sulforaphane, a chemical found cruciferous vegetables is an HDAC inhibitor, increases acetylated histone levels and has anti-cancer properties in the prostate. Sulforaphane is an isothiocyanate found in cruciferous vegetables such as broccoli. This anticarcinogen was first identified as a potent inducer of Phase 2 enzymes, but evidence is mounting that SFN acts through other cancer chemopreventive mechanisms. We recently reported on a novel mechanism of chemoprotection by sulforaphane in human colon cancer cells and prostate epithelial cells, namely the inhibition of histone deacetylase (HDAC). We have also found that sulforaphane specifically targets HDAC3 and HDAC6 in prostate cancer cells, but not normal cells. This work suggests that this phytochemical may have the ability to alter epigenetic events that otherwise lead to disease prevention. In human trials, we have directly compared the effects of the whole food (broccoli sprouts) to commercially available supplements. We have found a significant decrease in bioavailability and impact on HDACs with supplements compared to the whole food. We believe the decreases in activity are due to lack of myrosinase release of sulforaphane from its glucosinolate precursors in the supplement. We have also performed new studies examining the effects of tea catechins on autoimmune responses, which were completed in collaboration with Bruno (UConn). Regulatory T cells (Treg) are critical in maintaining immune tolerance and suppressing autoimmunity. The transcription factor Foxp3 serves as a master switch that controls the development and function of Treg. We have found that EGCG both in vitro and in vivo can induce Foxp3 and increase Treg frequencies to cause immunosuppression. The mechanisms may be related to demethylation of the Foxp3 promoter. Purdue University (Connie Weaver). During the past reporting period, we have conducted several studies in animal models and humans to investigate the effect of dietary constituents thought to be beneficial for bone health by either enhancing calcium absorption or suppressing bone resorption. In both growing rats and humans we tested the effect of fibers including GOS and FOS on calcium absorption. We continued our studies of the effects of phytoestrogens on preventing bone loss in an OVX rat model and the effect of equol producing status on bone resorption in response to soy in postmenopausal women. 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 methylenetetrahydrofolate 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 (Richard Bruno and Sung Koo). We have been actively investigating the hepatoprotective actions of green tea on liver injury associated with obesity-induced nonalcoholic steatohepatitis. We conducted a green tea extract (GTE) intervention study in obese rodents during the past reporting period. Wistar rats (16-wk old, n=63) were fed a low-fat (LF; 10% kcal) diet containing no GTE or a high-fat (HF; 60% kcal) diet containing 0, 1, or 2% GTE for 8 wks. We then examined whether GTE reduces inflammation associated with NASH by regulating COX-2. Rats fed HF diets developed liver steatosis and had greater (P<0.05) hepatic malondialdehyde (MDA) and serum alanine aminotransferase (ALT). GTE normalized the levels of hepatic lipid, MDA, and ALT to those of LF controls. The activity and protein expression of hepatic COX-2 were greater in HF fed controls. GTE normalized COX-2 activity and decreased its expression below the levels of LF controls. Arachidonic acid (AA) was elevated in the total lipid, phospholipid (PL), and free fatty acid (FFA) fractions of hepatic lipid extract. Although GTE reduced total AA by 6-8% (P<0.05), the concentrations of AA in the PL and FFA fractions remained unaffected as was cytosolic phospholipase A2 activity. HF increased cytochrome P450 2E1 mRNA expression, whereas GTE did not affect its expression. Thus, data suggest that GTE reduces dietary fat-induced hepatic inflammation and oxidative stress by decreasing the activity and expression of COX-2, without altering the flux of AA between membrane PL and FFA pools. Additional studies are warranted to define whether GTE suppresses COX-2 mediated prostaglandin generation. In separate studies, studies led by Dr. Bruno have examined the bioavailability and pharmacokinetics of quercetin in humans. We enrolled otherwise healthy obese men and postmenopausal women (n = 4M/5F; 55.9 ± 6.4 y; 30.8 ± 4.1 kg/m2) to complete a randomized, cross-over study. Participants ingested quercetin (1095 mg) on 3 occasions with a fat-free (<0.5 g), low-fat (4 g), or high-fat (15 g) meal. Plasma was obtained prior to (0 h) and after quercetin ingestion (1, 2, 3, 4, 6, 8, 10, 12, and 24 h) to measure quercetin and its methylated metabolites isorhamnetin and tamarixetin using HPLC-Coularray. Compared to the fat-free trial, quercetin bioavailability increased by 32% (AUC0-24 h; P<0.05) during the high-fat trial only. The high-fat meal also increased maximum quercetin concentrations (Cmax; 1.60 ± 0.88 ¼M) by 45%, but time to maximum concentration (Tmax; 353 ± 105 min) and half-life (t1/2; 515 ± 144 min) were unaffected. Isorhamnetin AUC0-24 h increased by 19% and Cmax increased by 40% (0.24 ± 0.11 vs. 0.17 ± 0.06 ¼M) without affecting Tmax (486 ± 113 min) or t1/2 (1154 ± 633 min). Tamarixetin AUC0-24 h increased by 43% and Cmax by 46% (0.52 ± 0.29 vs. 0.36 ± 0.11 ¼M) without affecting Tmax (399 ± 110 min) or t1/2 (299 ± 287 min). Thus, dietary fat improves quercetin bioavailability, and subsequent biotransformation, by increasing its absorption. Further work is warranted to define the optimal amount and type of fat to improve quercetin bioavailability and the extent to which the bioactivity of quercetin is exerted through its methylated metabolites. University of Nebraska (Janos Zempleni) has been examining the biotinylation of histones mediated by holocarboxylase synthetase (HCS), which is an important mechanism to repress retrotransposons. De-repression of retrotransposons impairs genome stability and increases cancer risk. In previous studies we showed that both biotin and HCS deficiency decreases the enrichment of biotinylated histones at retrotransposons, leading to increased transcription of transposons, increased frequency of retrotranspositions, and increased incidence of chromosomal abnormalities. In these previous studies we also demonstrated that histone biotinylation marks co-localize with methylated cytosines and lysine-9-dimethylated histone H3 (H3K9me2), suggesting epigenomic synergies among these chromatin marks. We tested the hypothesis that the epigenomic synergies are mediated by physical interactions among HCS, the DNA methyl transferase DNMT1, the methyl-CpG-binding protein 2 (MeCP2), and the histone H3 methyltransferase EHMT-1. Analysis by yeast-two-hybrid assays, co-immunoprecipitation, and limited proteolysis assays are consistent with the theory that HCS interacts with DNMT1, MeCP2, and EHMT-1, and that these interactions might be responsible for the roles of biotin and methyl donors such as folate in genome stability and gene regulation. The common interests in epigenetic modification by dietary constituents is an area that continues to be examined in collaboration with Ho (Oregon State).

Peer-Reviewed Publications 1. Bao B, Rodriguez-Melendez R, Wijeratne SSK, Zempleni J. Biotin regulates the expression of holocarboxylase synthetase in a miR-539 pathway in HEK-293 human embryonic kidney cells. J Nutr 140:1546-1551, 2010 2. Bao B, Pestinger V, Hassan YI, Borgstahl GEO, Kolar C, Zempleni J. Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18. J Nutr Biochem (in press) 3. Bao B, Rodriguez-Melendez R, Zempleni J. Cytosine methylation in miR-153 gene promoters increases the expression of holocarboxylase synthetase, thereby increasing the abundance of histone H4 biotinylation marks in HEK-293 human kidney cells. J Nutr Biochem (in press) 4. Bowman GL, Shannon J, Ho E, Traber MG, Frei B, Oken BS, Kaye JA, Quinn JF (2010). Reliability and Validity of Food Frequency Questionnaire and Nutrient Biomarkers in Elders With and Without Mild Cognitive Impairment. Alzheimer Dis Assoc Disord. 25(1):49-57. 5. Carter TC, Pangilinan F, Troendle JF, et al. Evaluation of 64 candidate single nucleotide polymorphisms as risk factors for neural tube defects in a large Irish study population. Am J Med Genet A 2011;155A(1):14-21. doi: 10.1002/ajmg.a.33755 [doi]. 6. Chandra Christopher L, Lucas EA, Clarke SL, Smith BJ, Kuvibidila S. White button and shiitake mushrooms reduce the incidence and severity of collagen-induced arthritis in dilute brown non-agouti mice. J Nutr 2011;141(1):131-136. 7. Cheong J, Gunaratna N, McCabe G, Jackson G, Kempa-Steczko A, Weaver C. Bone seeking labels as markers for bone turnover: Validation of urinary excretion in rats. Osteoporosis Intl. 2010 DOI: 10.1007/s00198-010-1281-7. 8. Chitchumroonchokchai C., Ferruzzi M., Campbell W., Failla M. Dietary fats with increased ratio of unsaturated to saturated fatty acids enhance absorption of carotenoids and vitamin E by increasing both efficiency of micellarization and lipoprotein secretion. FASEB J. 2010; 24:539. 9. Chung M-Y, Yeung SF, Park HJ, Volek JS, Bruno RS. (2010). Dietary ±- and ³-Tocopherol Protect Against LPS-triggered hepatic injury in spontaneously obese mice. J Nutr Biochem, e-Pub Feb 4, 2010. 10. Clarke, JD, Hsu, A., Yu, Z, Dashwood, RH and Ho, E. (2011) Differential effects of sulforaphane on histone deacetylases, cell cycle arrest and apoptosis in normal prostate cells versus hyperplastic and cancerous prostate cells. Mol Nutr Food Res, 2011 Mar 4. doi: 10.1002/mnfr.201000547. [Epub ahead of print] 11. Ferruzzi MG, Lobo JK, Janle E, Cooper B, Simon JE, Wu Q-L, Welch C, Ho L, Weaver C, Pasinetti GM. Bioavailability of gallic acid and catechins from grape seed polyphenol extract is improved by repeated dosing in rats: implications for treatment of Alzheimer's Disease. J Alzheimers Dis 18:113-24, 2009. 12. Filenko NA, Kolar C, West JT, Hassan YI, Borgstahl GEO, Zempleni J, Lyubchenko YL. The role of histone H4 biotinylation in the structure and dynamics of nucleosomes. PLoS ONE 6:e16299, 2011 13. Flögel A, Kim D-O, Chung S-J, Song WO, Fernandez M-L, Bruno RS, Koo SI, Chun OK. (2010). Development and validation of an algorithm to establish a total antioxidant capacity database of U.S. diet. Int J Food Sci Nutr, 61(6):600-23. 14. Getz J, Lin D, Medeiros DM. 2011. The cardiac copper chaperone proteins Sco1 and CCS are up-regulated, but Cox1 and Cox4 are down-regulated, by copper deficiency. Biol. Trace El. Res. In press. 15. Guo Y, Bruno RS. (2011). Vasoprotective activities of quercetin. AgroFood Industry Hi-Tech, In Press. 16. Hassan YI, Moriyama H, Zempleni J. The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. Arch Biochem Biophys 495:35-41, 2010 (an image from this article was used for the journals cover page) 17. Ho, E and Dashwood, RH. (2010) Dietary manipulation of histone structure and function. World Rev Nutr Diet. 2010;101:95-102. 18. Hord, NG, Ghannam, J, Garg, JK, Pamela D. Berens, PB and Bryan NS. (2010) Nitrate and nitrite content of human, formula, bovine, and soy milks: implications for dietary nitrite and nitrate recommendations. Breastfeeding Medicine Oct 19. [Epub ahead of print]. 19. Hsu, A., Bray, TM and Ho, E. (2010) Anti-inflammatory effects of soy and tea in prostate cancer prevention. Exp. Biol. Med; 235(6):659-67. 20. Hsu, A., Bray, TM, Helferich, WG, Doerge, D. and Ho, E. (2010) Differential effects of whole soy extract and soy isoflavones on apoptosis in prostate cancer cells Exp. Biol. Med. 235(1): 9097. 21. Hsu, A., Bruno, R.S., Lohr, C.V., Dashwood, R.H., Bray, T.M., and Ho, E. (2010) Dietary soy and tea mitigate chronic inflammation and prostate cancer via NFkB pathway in the Noble rat model, in J. Nutr. Biochem 2010 Aug 27. [Epub ahead of print] 22. Song Y, Elias V, Loban A, Scrimgeour AG, Ho E. (2010) Marginal zinc deficiency increases oxidative DNA damage in the prostate after chronic exercise. Free Radic Biol Med. 48:82-88. 23. Janle E, Lila MA, Grannon, MD, Wood L, Higgins A, Yousef GG, Rogers RB, Kim H, Jackson GS, and Weaver C. Method for evaluating the potential of 14C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometry. Nuclear Instruments and Methods in Physics Research B, 268:1313-1316, 2010. 24. Janle EM, Lila MA, Grannan M, Wood L, Higgins A, Yousef GG, Rogers RB, Kim H, Jackson GS, Ho L, Weaver CM. Pharmacokinetics and tissues distribution of 14C labeled grape polyphenols in the periphery and the central nervous system following oral administration. J Med Food 13(4)926-33, 2010. 25. Kaur Mall G, Chew YC, Zempleni J. The mechanisms of biotin homeostasis are qualitatively similar but quantitatively different in human Jurkat lymphoid and HepG2 liver cells. J Nutr 140:1086-1092, 2010 26. Klein MA, Nahin RL, Messina MJ, Rader JI, Thompson LU, Badger TM, Dwyer JT, Kim YS, Pontzer CH, Starke-Reed PE, and Weaver C. Guidance from an NIH Workshop on Designing, Implementing, and Reporting Clinical Studies of Soy Interventions. J Nutr 140:1192S-1204S, 2010. doi:10.3945/jn.110.121830. 27. Kuiper HC, Bruno RS, Traber MG, Stevens JF. (2011). Vitamin C supplementation lowers urinary levels of 4-hydroperoxy-2-nonenal metabolites in humans. Free Rad Biol Med, 50(7):848-53. 28. Lawrance CC, Lucas EA, Clarke SL, Smith BJ, Kuvibidila S. Differential effects of isoflurane and CO2 inhalation on plasma levels of inflammatory markers associated with collagen-induced arthritis in DBA mice. Int Immunopharmacol. 2009;9(7-8):807-9. 29. Lucas EA, Li W, Peterson SK, Brown A, Kuvibidila S, Perkins-Veazie P, Clarke SL, Smith BJ. Mango modulates body fat and plasma glucose and lipids in mice fed high fat diet. British Journal of Nutrition in press 30. Lucas EA, Mahajan SS, Soung DY, Lightfoot SA, Smith BJ, Arjmandi BH. Flaxseed but not flaxseed oil prevented the rise in serum cholesterol due to ovariectomy in the golden Syrian hamsters. J Med Food, 2011;14(3):261-7. 31. Montrose DC, Horelik NA, Madigan JP, Stoner GD, Wang L-S, Bruno RS, Park HJ, Giardina C, Rosenberg DW. (2010). Anti-inflammatory effects of lyophilized black raspberry powder in ulcerative colitis. Carcinogenesis, 32(3):343-50. 32. Mun JG, Grannan MD, Lachcik PJ, Rogers RB, Yousef GG, Grace MH, Janle EM, Wu QL, Simon JE, Weaver CM, Lila MA. Tracking deposition of a 14C-radiolabeled kudzu hairy root-derived isoflavone-rich fraction into bone. Exp. Biol. Med 235:1224-1235, 2010. 33. Mun JG, Grannan M, Lachcik P, Reppert A, Yousef GG, Rogers RB, Janle EM, Weaver CM, Lila MA. Metabolic tracking of 14C-labeled isoflavones. Br J Nutr 9:1-8, 2009. 34. Ortiz, D., Pico, S., Pachon, H., Chitchumroonchokchai, C., Failla. M. Evaluation of bioaccessibility of health promoting compounds from beans, cassava, sweet potato and alfalfa foliar extracts. XXIX Latinoamerican Congress of Chemistry. Cartagena, Columbia. September 27-October 1, 2010. 35. Park HJ, Bruno RS. (2010). Hepatoprotective Activities Of Green Tea In Nonalcoholic Fatty Liver Disease. AgroFOOD Industry High-tech. (Invited Review), 21(1): 37-40. 36. Park HJ, Davis SR, Liang H-Y, Rosenberg DW, Bruno RS. (2010). Chlorogenic acid differentially alters hepatic and small intestinal thiol redox status without protecting against azoxymethane-induced colon carcinogenesis in mice. Nutr Cancer, 62(3):362-370. 37. Park HJ, DiNatale DD, Chung M-Y, Lee J-Y, Koo SI, Bruno RS. (2011). Green Tea Extract Attenuates Hepatic Steatosis by Decreasing Adipose Lipogenesis and Enhancing Hepatic Antioxidant Defenses in ob/ob Mice. J Nutr Biochem, 22(4):393-400. 38. Park HJ, Mah E, Bruno RS. (2010). Validation of HPLC-Boron Doped Diamond Detection for Assessing Hepatic Glutathione Redox Status. Anal Biochem, 407(2):151-9. 39. Park J, Marjani SL, Lai L, Samuel M, Wax D, Davis SR, Bruno RS, Prather RS, Yang X, Tian XC. (2010). Altered Gene Expression Profiles in the Brain, Kidney, and Lung of Deceased Neonatal Cloned Pigs. Cell Reprogram, 12(5):589-97. 40. Pestinger V, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. The histone biotinylation marks H3K9bio, H3K18bio, and H4K8bio are enriched in repeat regions and participate in the repression of transcriptionally competent genes in human primary fibroblasts and Jurkat lymphoblastoma cells. J Nutr Biochem (in press) 41. Pietrzik K, Bailey L, Shane B. Folic acid and L-5-methyltetrahydrofolate: comparison of clinical pharmacokinetics and pharmacodynamics. Clin Pharmacokinet 2010;49(8):535-48. doi: 10.2165/11532990-000000000-00000 [doi] 42. Polara-Cabrera K, Huo T., Schwartz S.J., Failla, M.L. Digestive stability and transport of norbixin, a 24-carbon carotenoid, across monolayers of Caco-2 cells. J. Agr. Food Chem., 2010; 58:5789-5794. 43. Rajendran, P, Williams, DE, Ho, E and Dashwood, RH. (201x) Role of metabolism in generating HDAC inhibitors. Crit Rev Biochem Mol Bio, in press 44. Reinwald, S., Mayer, L.P., Hoyer, P.B., Turner, C.H., Barnes, S., Weaver, C.M. A longitudinal study of the effect of genistein on bone in two different murine models of diminished estrogen-poducing capacity. J. Osteoporosis. 2010 45. Rendina E, Lim YF, Marlow D, Wang Y, Clarke SL, Kuvibidila S, Lucas EA, Smith BJ. Dietary supplementation with dried plum prevents ovariectomy-induced bone loss in C57BL/6 mice and modulates the immune response. J Nut Biochem, 2011 Mar 15. [Epub ahead of print] 46. Rios-Avila L, Prince SA, Wijeratne SSK, Zempleni J. A 96-well plate assay for high-throughput analysis of holocarboxylase synthetase activity. Clin Chim Acta 412:735-739, 2011 47. Rios-Avila L, Pestinger V, Wijeratne SSK, Zempleni J. K16-biotinylated histone H4 is overrepresented in repeat regions and participates in the repression of transcriptionally competent genes in human Jurkat lymphoid cells. (submitted) 48. Simpson JL, Bailey LB, Pietrzik K, Shane B, Holzgreve W. Micronutrients and women of reproductive potential: required dietary intake and consequences of dietary deficiency or excess. Part I--Folate, Vitamin B12, Vitamin B6. J Matern Fetal Neonatal Med 2010;23(12):1323-43. doi: 10.3109/14767051003678234 [doi]. 49. Simpson JL, Bailey LB, Pietrzik K, Shane B, Holzgreve W. Micronutrients and women of reproductive potential: required dietary intake and consequences of dietary deficiency or excess. Part II--vitamin D, vitamin A, iron, zinc, iodine, essential fatty acids. J Matern Fetal Neonatal Med 2011;24(1):1-24. doi: 10.3109/14767051003678226 [doi]. 50. Singh D, Pannier AK,* Zempleni J.* Identification of holocarboxylase synthetase chromatin binding sites using the DamID technology. *Contributed equally to this paper. Anal Biochem (in press) 51. Song Y, Elias V, Wong CP, Scrimgeour AG and Ho E. (2010) Zinc transporter expression profiles in the rat prostate following alterations in dietary zinc. Biometals 23:51-58. 52. Wijeratne SSK, Camporeale G, Zempleni J. K12-biotinylated histone H4 is enriched in telomeric repeats from human lung IMR-90 fibroblasts. J Nutr Biochem 21:310-316, 2010 53. Wong, CP, Magnusson, KR and Ho, E. (2010) Aging is associated with altered dendritic cell subset distribution and impaired proinflammatory cytokine response. Exp Gerontol 45:163-169. 54. Zempleni J, Li Y, Xue, J, Cordonier E. The role of holocarboxylase synthetase in genome stability is mediated partly by epigenomic synergies between methylation and biotinylation events. Epigenetics (in press) 55. Zhou J, Wang D, Schlegel V, and Zempleni J. Development of an internet based system for modeling biotin metabolism using Bayesian networks. Comp Methods Progr Biomed (in press) Books and Book Chapters 1. Zempleni J, Wijeratne SSK, Kuroishi T. Biotin. In: Present Knowledge in Nutrition. Erdman J, Macdonald I, Zeisel S (eds.), 10th edition. International Life Sciences Institute, Washington, DC, 2012 (in press) 2. 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 (submitted) 3. Bagley P, Shane B. (2010). Folate. In Encyclopedia of Dietary Supplements, Coates P, Betz JM, Blackman MR, Cragg GM, Levine M, Moss J, White JD, eds., 2nd ed., pp. 288-77, Informa Healthcare, New York. 4. Hord, NG (2011) 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, In Press. 5. Medeiros DM, Bono E. The Iron factor in bone development. IN: J. Anderson, Calcium and Phosphorus in Health and Disease, CRC Press, Boca Raton, In press. 6. Lucas EA, Dumancas GG, Smith BJ, Clarke SL, Arjmandi BH. Health benefits of bitter melon (Momordica charantia). In Bioactive Foods in Promoting Health: Fruits and Vegetables. Edited by Ronald Ross Watson and Victor R. Preedy (in press). Dissertations 1. Chung, Min-Yu. Hepatoprotection of Vitamin E and Green Tea on Oxidative Stress and Inflammatory Responses In Animal Models of Obesity-Triggered Nonalcoholic Fatty Liver Disease. PhD Thesis 2011. 2. Hill, Katherine. Predictors of skeletal calcium accretion in adolescents from pooled metabolic balance studies. PhD. Thesis, 2010. 3. Legette, LeeCole. Effect of soy isoflavones and Prebiotics on bone metabolism in a postmenopausal rodent model. PhD. Thesis, 2010. Abstracts 1. KD Ballard, BR Kupchak, BM Volk, DJ Friedenreich, JC Aristizabal, E Mah, Y Guo, C Masterjohn, RS Bruno, WJ Kraemer, JS Volek. (2010). A 9-month progressive resistance training program attenuates acute exercise-induced lipid peroxidation in healthy men and women. American College of Sports Medicine (Abstract). 2. M-Y Chung, HJ Park, JE Manautou, SI Koo, RS Bruno. (2010). Green tea extract protects against nonalcoholic steatohepatitis in ob/ob mice by decreasing oxidative and nitrative stress responses induced by pro-inflammatory enzymes. Northeast Chapter of the Society of Toxicology; Storrs, CT. Abstract. 3. Clarke, JD, Yu, Z, and Ho, E (2010). Differential effects of sulforaphane on histone deacetylases, cell cycle arrest and apoptosis in normal and prostate cancer cells FASEB J. 2010 24:107.5 4. Ho, E, Hardin, K., Wong, CP,Hobson, B, Traber, MG and Tanguay, RL (2010) Using zebrafish as a model organism to understand the impact of maternal zinc status on embryonic zinc homeostasis during development FASEB J. 2010 24:718.10 5. Kuroishi T, Cerny RL, Zempleni J. Mass spectrometric analysis of biotinylated peptides and histones. Abstract 3809 (107.2) Experimental Biology Meeting 6. A Larson, Y Guo, KS Black, MA Hayman, RS Richardson, RS Bruno, T Jalili, JD Symons. (2010). A single dose of quercetin lowers blood pressure in hypertensive but not prehypertensive males. Experimental Biology, Anaheim, CA. Abstract. 7. E Mah, K Ballard,D Kawieki, JS Volek, RS Bruno. (2010). ³-Tocopherol supplementation improves postprandial vascular endothelial function in lean and obese men by decreasing oxidative and nitrative stress. Experimental Biology, Anaheim, CA. Abstract. 8. HJ Park, M-Y Chung, SI Koo, RS Bruno. (2010) Green tea extract (GTE) attenuates hepatic and adipose inflammation by altering glutathione redox status in nonalcoholic fatty liver disease (NAFLD). Experimental Biology, Anaheim, CA. Abstract. 9. HJ Park, M-Y Chung, SI Koo, Y-K Park, J-Y Lee, RS Bruno. (2010) Green tea extract protects against nonalcoholic fatty liver disease in diet-induced obese rats by decreasing hepatic inflammatory and oxidative stress responses. Institute of Food Technologists; Chicago, IL. Abstract. 10. Rodriguez-Melendez R, Bui DC, Ellis M, Johnson RM, Zempleni J. (2010) Identification of a potential role of K9-biotinylated histone H3 in honeybee (Apis mellifera) development. Abstract 3220 (550.2) Experimental Biology Meeting 11. Rios-Avila L, Wijeratne SSK, Pestinger V, Zempleni J. (2010) Characterization of the H4K16bio mark in human lymphoid cells. Abstract 2273 (550.1) Experimental Biology Meeting 12. Rios-Avila L, Pestinger V, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. Characterization of the H4K16bio mark in human cells. UNL Research Fair, April 7, 2010, Lincoln, NE 13. Sands, D.C., 2011. The quest for gluten-free grains more nutritious than wheat. International Celiac Disease Symposium. (Abstract). Oslo, Norway, 2011. 14. K Sankavaram, L Chong, RS Bruno, HC Freake. (2010). Zinc deficiency induces apoptosis but zinc induces necrosis in rat hepatoma cells. Experimental Biology, Anaheim, CA. Abstract. 15. Singh D, Zempleni J, Pannier A. Development of an Antibody Independent Technology to Monitor Chromatin Proteins in Cell Lines. University of Nebraska Medical Center, Regenerative Medicine Symposium, May 24, 2010, Omaha, NE 16. Tian XC, Park J, Bruno R, French RA, Prather RS. 2010. Functional genomics and epigenetics in pig cloning research. 43rd Annual Meeting of the Society for the Study of Reproduction; July 30-Aug 3, 2010; Milwaukee, WI. Biol Reprod 83 (S1):22 (Abstract 104). 17. Wijeratne SSK, Kuroishi T, Pestinger V, Rios-Avila L, Zempleni J. (2010) Histone biotinylation is a naturally occurring phenomenon. Abstract 2316 (107.1) Experimental Biology Meeting Other 1. Lucas EA, Peterson SK, Clarke SL, and Smith BJ. Cardiovascular health benefits of grape juice. Handbook of Oklahoma Vineyard Establishment and Management ed. Stafne ET (Oklahoma Cooperative Extension Service Circular E-1015, 2010).