Brief Summary of Minutes of Annual Meeting

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.

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.

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). 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

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 http://www.hindawi.com/journals/ecam/2016/8135135/

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. http://dx.doi.org/10.3389/fnut.2016.00004. eCollection 2016.

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 http://jn.nutrition.org , 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