Brief Summary of Minutes of Annual Meeting

The 2016 Annual Meeting of the W-3122 Multistate Group was called to order by the Chair, Dr. Nancy Turner on the morning of October 6. Dr. Mike Harrington (WAAESD) provided an overview of the status of the multi-state program, the current funding priorities identified in the NIFA strategic plan, and budget status at NIFA. Last year, this group was recognized with the Western Region Award of Excellence in Multistate Research. Dr. Harrington would like to resubmit a nomination for W3122 to be considered for the national award again this year. Each member present delivered a presentation to update the group on his/her progress (content summarized in the Annual Group Report). At the business meeting, new officers for 2016-2017 were selected: Chair, Meijun Zhu; Vice-Chair, Yuhua Farnell; and Secretary, Susan Tilton. Also, the dates for the 2017 annual meeting will be October 4-6 and the meeting will be held in Calistoga, CA.

Reports submitted, but not able to attend annual meeting

Benninghoff, Abby, abby.benninghoff@usu.edu (Utah State University)

Denison, Michael, msdenison@ucdavis.edu (University of California, Davis)

Eisenstein, Richard, eisenste@nutrisci.wisc.edu (University of Wisconsin-Madison)

Friedman, Mendel, friedman@ars.usda.gov (USDA-ARS Albany)

Nerurkar, Pratibha, pratibha@hawaii.edu (University of Hawaii)

Pagliarini, David, pagliarini@morgridge.org (Morgridge Institute for Research)

Pestka, Jim, pestka@msu.edu (Michigan State University)

Ricketts, Marie-Louise, mricketts@cabnr.unr.edu (University of Nevada, Reno)

Romagnolo, Donato, donato@u.arizona.edu (University of Arizona)

Weir, Tiffany, tiffany.weir@colostate.edu (Colorado State University)

OBJECTIVE 1: Determine the mechanisms by which dietary bioactive compounds protect against human diseases.

Dr. Mendel Friedman (USDA-ARS Albany)

Dr. Mendel Friedman and colleagues have demonstrated that combinations of shiitake mushroom (Lentinus edodes) mycelia with other sources of bioactive compounds, such as fermented elm tree (Ulmus parvifolia) bark extract or turmeric are protective against several disease states. In the first study, they found the combination of elm tree bark extract and the shiitake mushroom mycelia culture resulted in anti-inflammatory and anti-allergic actions in a mouse model of allergic asthma. In the second study, they found the combination of turmeric and the shiitake mushroom mycelia culture protected mice against both liver damage (necrosis) and lethality induced by Salmonella Typhimurium. The protection was induced by stimulation of the immune system.

Dr. Roger Coulombe (Utah State University)

Dr. Roger Coulombe and colleagues have developed an avian embryo exposure model that reduces use of live animals in research, and allows them to study the molecular mechanisms of regulatory pathways of the universal detoxifying enzyme glutathione S-transferases. Previous results from our laboratory have demonstrated that domesticated turkeys (Meleagris gallopavo) are especially sensitive to aflatoxicosis, while Eastern wild turkeys (M. g. silvestris) are relatively resistant. In ovo exposure provided a controlled AFB1 challenge and comparison of domesticated and wild turkeys. Gene expression responses to AFB1 in the embryonic hepatic transcriptome were examined using RNA-sequencing (RNA-seq). Eggs were injected with AFB1 (1 g) or sham control and dissected for liver tissue after 1 day or 5 days of exposure. Libraries from domesticated turkey (n = 24) and wild turkey (n = 15) produced 89.2 Gb of sequence. Approximately 670 M reads were mapped to a turkey gene set. Differential expression analysis identified 1535 significant genes with |log2 fold change| > 1.0 in at least one pair-wise comparison. AFB1 effects were dependent on exposure time and turkey type, occurred more rapidly in domesticated turkeys, and led to notable up-regulation in cell cycle regulators, NRF2-mediated response genes and coagulation factors. Further investigation of NRF2-response genes may identify targets to improve poultry resistance.

Changes in methylation, particularly at CpG sites clustered in the promoter region of a gene, can lead to changes in expression. To identify whether methylation differences may be responsible for GST silencing in domesticated turkeys, Dr. Coulombe’s group searched for CpG islands "upstream" of the 5' end GSTA genes. CpG sites in the GSTA region occur less frequently than expected by chance, with apparent CpG islands near the 5' ends of most of the GSTA genes, but not GSTA3--the one we are most interested in. The SNPs that have been identified in this region between wild and domestic turkeys do not affect the number of CpG sites, so it doesn't appear that a CpG island which was present in wild turkeys has been lost in domestic turkeys.

Genes may share promoters, so we are examining the CpG island in the promoter of GSTA4 (the next gene upstream from GSTA3). The Coulombe lab has designed primers to amplify the GSTA4 promoter region and will be using a methylation sensitive restriction enzyme-based method to determine whether the CpG sites are differentially methylated between Wild and Domestic turkeys.

Dr. Abby Benninghoff (Utah State University)

Dr. Benninghoff’s group reported preliminary results from an ongoing multi-generation mouse study designed to determine the impact of ancestral or multi-generational consumption of the total Western diet (TWD), a Western-style diet formulated for rodents using human US nutrient intake data, in a murine model of inflammation-associated colorectal carcinogenesis.  C57BL/6J mice were bred for three generations, during which they were fed an optimized diet (AIN93G), TWD or a simple high fat diet (45% DIO) in the F₀ only, F₀ through F₃, or the F₃ generation only.  The azoxymethane and dextran sodium sulfate model of inflammation-associated CRC was employed in the F₃ offspring.  Notably, ancestral exposure (F₀ generation) to the TWD markedly enhanced CRC incidence and disease severity as measured by tumor multiplicity and burden in F₃ offspring (P<0.05), whereas the DIO diet had little effect on colon tumorigenesis (P>0.05).  Moreover, cumulative exposure to TWD over multiple generations had an even stronger effect on CRC severity compared to F₃ offspring provided TWD directly (P<0.05).  Ancestral exposure to either TWD or DIO diet did not appear to markedly alter body weight or composition in F₃ offspring (P>0.05).  These data suggest that ancestral exposure to a Western-style diet may be as detrimental as direct exposure in regards to colon tumorigenesis.  Also, as part of this study, another cohort of F₃ mice were provided drinking water supplemented with green tea extract, with the objective to determine how ancestral patterns of exposure to the Western diet impacted the efficacy of green tea as an anticancer agent.  Analyses of outcomes from this arm of the study are ongoing.

Previously, Dr. Benninghoff’s research group (USU) reported that dietary supplementation with tart cherries caused a significant 40% reduction in tumor incidence in mice fed AIN93G, whereas tart cherries had no effect on tumor incidence in mice fed TWD.  However, tart cherry powder supplementation did not significantly affect tumor multiplicity, tumor size or colitis in mice fed either basal diet. In ongoing analysis of this work, her group examined the impact of TWD and tart cherry supplementation on colon inflammation.  The azoxymethane and dextran sodium sulfate (DSS) model of inflammation-associated CRC was employed, which triggers colitis within 10 days of DSS treatment.  Disease activity, colon tissue histopathology and biomarkers of inflammation (immunohistochemistry) were assessed at multiple time points, including colitis, recovery and at the study end. Mice fed TWD experienced significantly elevated levels of inflammation and mucosal injury during the colitis phase, as compared to mice fed AIN93G, and this elevated response was evident throughout recovery and progression to tumorigenesis.  However, treatment with tart cherry did not significantly affect inflammation or mucosal injury.  Thus, the anticancer activity of tart cherry is likely not related to changes in colonic inflammation.

Dr. Marie-Louise Ricketts (University of Nevada, Reno)

In an effort to determine the potential use of a grape seed procyanidin extract (GSPE) as a natural therapy to treat or ameliorate metabolic disorders, Dr. Ricketts and colleagues have conducted several in vitro and in vivo studies, in order to further elucidate its’ molecular mechanism of action. To date our investigations have led to the following findings:

GSPE exerts a triglyceride-lowering effect in a hyperlipidemic state

In this study, rats were fed either a starch control diet or a diet containing 65% fructose for 8 weeks to induce hypertriglyceridemia. During the 9th week of the study, rats were maintained on their respective diet and administered vehicle (water) or GSPE via oral gavage for 7 days. Fructose increased serum triglyceride levels by 171% after 9 weeks, compared to control. GSPE administration in the fructose-fed rats attenuated this effect, resulting in a 41% decrease. GSPE inhibited hepatic lipogenesis via down-regulation of several genes including, sterol regulatory element binding protein 1c (Srebp1c) and stearoyl-CoA desaturase 1 (Scd-1) in the fructose-fed animals. GSPE increased fecal bile acid and total lipid excretion, decreased serum bile acid levels and increased the expression of genes involved in hepatic cholesterol synthesis. However, hepatic bile acid biosynthetic gene expression was not increased in the presence of GSPE and fructose. Serum cholesterol levels remained constant, while hepatic cholesterol levels decreased. GSPE did not modulate expression of genes responsible for esterification or biliary export of the newly synthesized cholesterol, but did increase fecal cholesterol excretion, suggesting that in the presence of GSPE and fructose, the liver may secrete more free cholesterol into the plasma, which may then be shunted to the proximal small intestine for direct basolateral to apical secretion and subsequent fecal excretion. The results from this study demonstrate that GSPE effectively lowers serum triglyceride levels in fructose-fed rats after one-week administration. This study provides novel insight into the mechanistic actions of GSPE in treating hypertriglyceridemia and demonstrates that it targets hepatic de novo lipogenesis, bile acid homeostasis and non-biliary cholesterol excretion as important mechanisms for reducing hypertriglyceridemia and hepatic lipid accumulation in the presence of fructose, a highly lipogenic dietary component.

Dietary procyanidins alter enterohepatic bile acid recirculation: elucidation of a novel mechanism to reduce triglyceridemia

Understanding the molecular basis by which dietary procyanidins modulate triglyceride and cholesterol homeostasis has important implications for the use of natural products in the treatment and prevention of cardiovascular disease. To determine whether modulation of bile acid homeostasis contributes to the hypotriglyceridemic action of GSPE, the effect on genes regulating bile acid absorption, transport and synthesis was examined in vitro, in Caco-2 cells, and in vivo, in wild type (C57BL/6) and farnesoid x receptor knockout (Fxr-/-) mice. Results from this study provide novel evidence demonstrating that GSPE is a naturally occurring gene-selective bile acid receptor modulator (BARM). Mechanistically, GSPE down-regulates genes involved in intestinal bile acid absorption and transport in an FXR-dependent manner, resulting in decreased enterohepatic bile acid recirculation. This correlates with increased fecal bile output, decreased serum triglyceride and cholesterol levels, increased hepatic cholesterol 7α-hydroxylase (Cyp7a1), and decreased intestinal fibroblast growth factor 15 (Fgf15) expression. GSPE also increased hepatic HmgCoA reductase (Hmgcr) and synthase (Hmgcs1) expression, while concomitantly decreasing sterol regulatory element-binding protein 1c (Srebp1c). GSPE selectively regulates intestinal FXR-target gene expression in vivo, and modulation of bile acid absorption and transport is a critical regulatory point for the consequential hypotriglyceridemic effects of GSPE.

GSPE exerts complementary effects with respect to triglyceride-lowering when administrated in conjunction with the bile acid sequestrant, cholestyramine

Bile acid sequestrants are lipid-lowering agents, which may be prescribed as a monotherapy or combination therapy to reduce the risk of coronary artery disease. Over 33% of adults in the United States use complementary and alternative medicine strategies, and we recently reported that grape seed procyanidin extract (GSPE) reduces enterohepatic BA recirculation as a means to reduce serum triglyceride levels. This study was designed to assess the effects on bile acid, cholesterol and triglyceride homeostatic gene expression following co-administration with GSPE and the bile acid sequestrant, cholestyramine (CHY). Eight-week old male C57BL/6 mice were treated for 4 weeks with either a control or 2% CHY-supplemented diet, after which, they were administered vehicle (water) or GSPE for 14 hours. Liver and intestines were harvested and gene expression was analyzed. Bile acid, cholesterol, non-esterified fatty acid and triglyceride levels were also analyzed in serum and feces. Results revealed that GSPE treatment alone, and co-administration with CHY, regulates bile acid, cholesterol and triglyceride metabolism differently than CHY administration alone. Notably, GSPE decreased intestinal apical sodium-dependent bile acid transporter (Asbt) gene expression, while CHY significantly induced expression. Administration with GSPE or CHY robustly induced hepatic bile acid biosynthetic gene expression, especially cholesterol 7α-hydroxylase (Cyp7a1), compared to control, while co-administration further enhanced expression. Treatment with CHY induced intestinal and hepatic cholesterologenic gene expression, while co-administration with GSPE attenuated the CHY-induced increase in the liver but not intestine. CHY also induced hepatic lipogenic gene expression, which was attenuated by co-administration with GSPE. Consequently, a 25% decrease in serum triglyceride levels was observed in the CHY+GSPE group, compared to the CHY group. Collectively, this study presents novel evidence demonstrating that GSPE provides additive and complementary efficacy as a lipid-lowering combination therapy in conjunction with CHY by attenuating hepatic cholesterol synthesis, enhancing bile acid biosynthesis and decreasing lipogenesis, warranting further investigation.

GSPE inhibits HDAC activity leading to increased Pparα phosphorylation and target-gene expression to increase fatty acid ß-oxidation

Histone deacetylases (HDACs) have emerged as epigenetic regulators of risk factors associated with the metabolic syndrome (MetS), and certain botanical extracts have proven to be potent HDAC inhibitors. Understanding the role of dietary procyanidins in HDAC inhibition is important in exploring the therapeutic potential of natural products. In this particular study, C57BL/6 mice were gavaged with vehicle (water) or grape seed procyanidin extract (GSPE, 250 mg/kg) and terminated 14 h later. Liver and serum were harvested to assess the effect of GSPE on HDAC activity, histone acetylation, Pparα activity and target-gene expression, and serum lipid levels. Results show that GSPE increased histone acetylation and decreased Class I HDAC activity in vivo, and dose-dependently inhibited recombinant HDAC2 and 3 activities in vitro. Accordingly, Pparα gene and phosphorylated protein expression were increased, as were target genes involved in fatty acid catabolism, suggesting increased Pparα activity. Serum fibroblast growth factor 21 (Fgf21) was elevated, and triglyceride levels were reduced by 28%. This study revealed that mechanistically GSPE regulates HDAC and Pparα activities to modulate lipid catabolism and reduce serum triglycerides in vivo.

Dr. Pratibha Nerurkar (University of Hawaii)

In Hawaii, minority populations such as Native Hawaiians and Pacific Islanders (NHPI) have more than twice the rate of obesity-associated type 2 diabetes (T2D) as Caucasians, and are more than five times as likely to die from T2D. Current therapies for obesity are complicated due to factors including an inability to maintain long-term weight loss and drug-drug interactions. In addition, conventional therapies may not be affordable, suitable and/or acceptable for culturally sensitive minority populations. There is a growing awareness and mounting body of scientific evidence, that successful implementation of strategies to control T2D among ethnic minorities will require culturally appropriate interventions.

Dr. Pratibha Nerurkar and colleagues have demonstrated that laboratory-prepared juice (fNJ) improves not only glucose and lipid metabolism, but also prevents weight gain in mice fed high fat diet (HFD, 58% fat). Although the worldwide market for noni products are currently estimated at $2 billion and laboratory-based research has supported health claims of noni, clinical studies supporting health benefits of noni are scant and limited to Tahitian noni juice. We compared the anti-diabetic and anti-inflammatory effects of fNJ and commercial noni juices, differences in chemical metabolites and modulation of gut microbiome. Our findings indicate that laboratory-based fNJ was more potent at improving glucose metabolism and inflammation within four weeks of starting treatment of HFD-fed mice. These effects are possibly associated with differences in the chemical composition of commercial and laboratory-prepared juices, as determined by metabolomics studies.

Dr. James Pestka (Michigan State University)

Lupus is a debilitating autoimmune disease that adversely affects 1.5 million Americans. While the genome is a primary predisposing factor for autoimmunity, lifetime environmental exposures to factors (i.e., exposome) such as environmental toxicants and diet are now recognized to modulate hereditary effects. Experimental animal and epidemiological studies have linked exposure to the respiratory toxicant silica to lupus and other autoimmune diseases. Importantly, nearly 2 million Americans are occupationally exposed to respirable silica. In contrast, both animal and clinical studies suggest that consumption of DHA and other ω-3 PUFAs can both prevent and resolve inflammation and autoimmune nephritis. Little is known about how respiratory toxicants like silica trigger lupus or how DHA supplementation could be harnessed to block environmental triggering. This critical barrier results in unnecessary lupus burden in genetically predisposed people that are exposed to respiratory toxicants. Lupus onset/progression in such individuals could potentially be mitigated by development and implementation of effective, low cost prophylactic approaches employing optimized supplementation with ω-3 PUFAs, already regularly consumed by over 20 million Americans. Research in the Pestka lab focuses on using such an approach to prevent lupus. Taken together, our research team now has a model that enables dissection of the countervailing roles of silica (potentiation) and DHA (attenuation) in lupus initiation.

Dr. Nancy Turner (Texas A&M University)

Colon cancer and inflammatory bowel disease are major contributors to morbidity and mortality in the US, and dietary patterns have been suggested to be responsible for a large fraction of the cases. Recent evidence demonstrates diet may influence these diseases through its impact upon the microbiota. We are exploring the impact of dietary fiber sources, particularly those with unique or high levels of phenolic compounds on colon health.

In our first experiment, we determined if polyphenol-rich sorghum brans would influence the colon microbiota of healthy rats. The experimental diets included a sorghum bran that contained either condensed tannins, deoxyanthocyanins or a combination of the two. Within four weeks of changing from a standard pelleted diet to the semi-purified experimental diets (devoid of phenolic molecules), rats consuming cellulose as the fiber source had an elevated proportion of Firmicutes and a dramatic reduced proportion of Bacteroidetes. In contrast, rats consuming a diet containing condensed tannins had reduced Firmicutes and elevated Bacteroidetes. Those consuming a diet containing either diet with deoxyanthocyanins maintained relatively similar proportions of these microbial phyla. These data demonstrate that diets low/depleted of polyphenols induces greatly elevated ratios of Firmicutes/Bacteroidetes, a pattern similar to that found in obese individuals or those with inflammatory bowel disease. Importantly, it was possible to protect against this shift by including a single, polyphenol-rich fiber source.

Our second project explored the impact of dried plums on colon cancer, where we found a 50% reduction in early colon lesions in rats consuming a diet containing dried plums. There were changes in the distal colon microbiota of carcinogen-injected rats consuming the control diet. However, the dried plum diet produced minimal changes in microbiota between the proximal and distal colon, and the changes in microbiota induced by the carcinogen were not as prominent in the dried plum group. We are performing untargeted metabolomics analyses on feces to determine if the changes in microbiota resulted in altered metabolite profiles. We discovered several compounds endogenous to plums in the luminal contents (both proximal and distal), with several of those compounds being demonstrated in the existing literature to suppress proliferation and enhance apoptosis in colon cancer cells. In addition, there were multiple compounds produced from microbial metabolism of phenolic molecules identified in the luminal contents from the proximal and distal colon of rats provided the plum diet. These molecules have also been demonstrated to protect against carcinogenesis. Additional experiments using dried plums, or compounds isolated from plums have demonstrated effects on bone, as well as colon cancer, demonstrating the potential for systemic benefits of dried plum consumption.

Dr. Tiffany Weir (Colorado State University)

Colorectal cancer is one of the major causes of cancer deaths, but the majority of cases can be prevented by modification of lifestyle factors such as diet. In particular, fiber intake has been associated with a decreased risk of developing colorectal cancer. Recently, several lines of evidence have suggested roles for the gut microbiota in development and progression of colorectal cancers, and dietary fiber is an important modulator of gut bacterial populations. We are testing the hypothesis that dietary intervention using two sources dietary fiber, stabilized rice bran (SRB) and navy bean powder (NBP), will modulate the gut microbiota and improve the intestinal environment for reduced risk of colorectal cancer. Toward this goal, we have identified specific microbial (and metabolite) changes associated with increased intake of either SRB or NBP in healthy cohorts with and without a history of colorectal cancer. Because human-associated microbiota responded differently between SRB and NBP treatment groups, ongoing research is aimed at specifically examining the different fiber composition of these foods alters their interactions with the microbiota. Specifically, we are exploring the interaction between arabinoxylans from SRB and how they interact with mucin-degrading bacteria such as Bacteroides ovatus and Akkermansia muciniphila.

High fruit and vegetable (F&V) consumption is also associated with protection against CRC as well as a number of other diseases. However, this protective effect cannot be explained by increased fiber intake alone and is likely due to the myriad phytochemicals present in these foods. To better understand how F&V consumption mechanistically affects host physiology for improved health outcomes, biomarkers of F&V intake are needed. To this end, we are exploring various metabolomic profiling strategies to identify chemical signatures associated with varying levels of F&V intake. Completed analysis of urine samples from a controlled F&V feeding study indicate that there are multiple metabolites associated with either high or low F&V intake, including many phytochemicals specific to particular botanical families (i.e., sulforaphane and capsaicin), as well as several host derived metabolites (such as sialic acid-the terminal molecule in intestinal mucins), which may serve as indicators of host health. Future studies will be directed at exploring ionomics platforms for signature generation as well as validating the biomarker signatures that have already been identified using a larger study population.

Dr. Richard Eisenstein (University of Wisconsin, Madison)

First, we are investigating the extent to which the pathologies associated with obesity and iron overload may partly act through similar signaling pathways. We are interested in understanding the basis of the mild anemia observed in obesity and the role of the so-called unfolded protein response (UPR) and signaling pathways down-stream of the UPR that may modulate production of the iron regulatory protein hepcidin. Evidence in the literature indicates that hepcidin expression is activated by the UPR. Activation of the UPR in obese subjects could stimulate hepcidin production, which is predicted to reduce dietary iron absorption and iron recycling leading to anemia. A final component of this work is to examine if expression of the transcription factor HIF2-alpha is altered in obese mice. HIF2alpha expression is controlled at the translational level by iron and we hypothesize a role for the UPR. Part of our work here focuses on defining the role of translational regulatory elements in the 5’ untranslated region of the HIF2alpha mRNA using a luciferase reporter assay.

Second, we are examining how the iron regulated RNA binding protein, iron regulatory protein 1 (IRP1), controls dietary iron absorption and erythropoiesis during fetal and postnatal development. An ultimate goal of this, and a subject of our recently submitted Hatch renewal proposal, is whether dietary factors known to control HIF2alpha expression might consequently control dietary iron absorption. IRP1 is the key iron mediator of HIF2alpha mRNA translation. IRP1 is a repressor of HIF2alpha synthesis and loss of IRP1 translationally activates HIF2alpha mRNA. HIF2alpha is a transcriptional activator of the main blood forming hormone erythropoietin and also the iron transport system controlling dietary iron acquisition in the duodenum. Mice lacking IRP1 develop a severe but transient polycythemia (too many red cells) and absorb more iron to promote red cell overproduction. We have recently found that these mice have very strongly repressed hepcidin mRNA at birth and our data obtained over the past year suggests that this does not involve canonical mechanisms for controlling hepcidin gene transcription. Furthermore, our studies over the past year indicate that IRP1 is an important factor controlling placental iron transfer from the mother to the fetus. We are currently writing a paper on this work.

Third, we have further examined the role of IRP1 in controlling HIF2alpha and erythropoiesis by using a zinc-finger nuclease to delete the iron responsive element (IRE) in HIF2alpha mRNA so that IRP1 cannot control translation of this message. Mice lacking the IRE in HIF2alpha display a dysregulation of erythropoiesis that appears milder, in some respects, than in mice lacking IRP1. We are focusing on what additional mechanisms may be dysregulated in mice lacking IRP1 vs. those lacking the IRE in HIF2alpha mRNA.

Fourth, we are examining the iron-dependent mechanisms controlling IRP1 function as a means to understand how dietary iron absorption is controlled in disease states including iron deficiency as well as in inflammatory disorders. This includes the known impact of inflammatory scenarios in reducing intestinal iron absorption. The ability of IRP1 to bind mRNA is determined by insertion or loss of an Fe-S cluster with the cluster-free apoprotein form binding RNA. A second mechanism involves iron-dependent degradation of the IRP1 apoprotein. To better understand how dietary iron absorption is controlled we are focusing on the ability of IRP1 to be regulated by the Fe-S cluster vs. the protein degradation mechanism. A manuscript on this work will be submitted in November 2016.

Fifth, our expertise on the regulation of function of IRP1 and iron metabolism in general has allowed us to collaborate with other labs study the control of cellular iron metabolism including the lack of impact of pharmacological agents on IRP1.

Dr. David Pagliarini (University of Wisconsin, Madison)

We have continued to investigate the effects of iron deprivation on the cellular control of mitochondrial biogenesis. Using microarray and quantitative mass-spectrometry approaches, we discovered that depriving C2C12 mouse myotubes of iron, through treatment with the iron chelator deferoxamine (DFO) leads to a global decrease in the transcript abundance of mitochondrial- and nuclear-encoded mitochondrial genes and mitochondrial proteins. We have found that this response to iron chelation is universal across a broad range of cell types, rapid (occurring within 24 hours) and dose-dependent. We also found that the effect of iron chelation on transcript levels is fully reversed with (histone deacetylase) HDAC inhibition, consistent with histone post-translational modification (PTM)-mediated regulation of gene expression. To buttress our observations on the role of histone-PTMs in the iron deprivation induced transcriptional regulation of mitochondrial biogenesis, we performed additional transcript labeling and ChIP-qPCR analyses. First, to test whether the change in acetylation is simply a general decrease found at all genes regardless of their expression, we added the mitochondrial non-OxPhos encoding gene Vdac1, whose transcript levels are not affect by iron chelation. We found that there is no change in Vdac1 transcript degradation, and we observed decreases in the synthesis of Vdac1 at the final two time points of our time-course, which occurred after the changes in OxPhos transcripts. We also only observed relatively small (and no statistically significant) changes in histone acetylation and no change in histone methylation at Vdac1 consistent with the lack of change in Vdac1 expression in response to iron chelation. We also added non-mitochondrial genes to our analyses, including Hk2 and Slc2a1 (both involved in glucose metabolism). We found that their transcription increases in response to DFO, and we also find that DFO increases their stability, suggesting that regulation at both levels (synthesis and degradation) contributes to the overall increase in their total transcript abundance. Our ChIP-qPCR analysis shown that iron deprivation causes no significant change in histone acetylation (or methylation) at either gene, suggesting that the loss of acetylation is specific to certain genes rather than genome-wide. Second, we added more mitochondrial OxPhos-encoding genes to our ChIP-qPCR analyses, including Ndufs8 and Cycs, and found that their histone-PTM profiles are consistent with the previously analyzed OxPhos-encoding genes.

Next, as we observed a general increase in methylation following iron deprivation, we used the demethylase inhibitor dimethyloxalylglycine (DMOG) to test if a decrease in histone demethylase activity is sufficient to cause a similar mitochondrial response. Most lysine demethylase reactions are catalyzed by the Jumonji C (JmjC)-domain containing demethlyases that require α-ketoglutarate, O₂ and Fe²⁺. These JmjC demethylases can be inhibited by a-ketoglutarate analogs, such as DMOG, without affecting iron levels. Interestingly, we found that DMOG did not affect OxPhos-encoding transcripts, suggesting that methylation is not the main regulator of the mitochondrial transcriptional response to iron deprivation. Finally, we published these recent discoveries on iron deprivation-induced, histone-PTM transcriptional regulation of mitochondrial biogenesis in the 2016 September 30 issue of the Journal of Biological Chemistry.

Dr. Meijun Zhu (Washington State University)

Beyond their nutritional impact on colonic epithelial cells, the intestinal microbiota metabolite butyrate has been shown to exert beneficial effects on intestinal homeostasis and metabolism. Mast cells are well known for their roles in mediating allergy responses and regulating gut inflammation. We demonstrate that butyrate profoundly inhibited proliferation of mouse mastocytoma P815 cells through inducing cell cycle arrest and apoptosis, as well as decreasing c-Kit activation; butyrate also increased early and late stage apoptotic P815 cells. Additionally, butyrate suppressed FceRI-dependent cytokine production in murine primary BMMC, which are likely mediated by inhibition of histone deacetylation.

Goji berry is a traditional Chinese tonic food. Dietary goji berry ameliorated DSS-induced body weight loss, diminished diarrhea and gross bleeding, resulted in a significantly decreased disease activity index, as well as DSS associated colon shortening. Histologically, it ameliorated mucosal damage and neutrophil infiltration into colonic intestinal tissue in response to DSS challenge, which was associated with decreased expression of chemokine (C-X-C motif) ligand 1 (CXCL1) and monocyte chemoattractant protein-1 (MCP-1), as well as inflammatory mediators, interleukin-6 and cyclooxygenase-2. In summary, Goji supplementation confers protective effects against DSS­induced colitis, which has the potential to be used as a complementary therapeutic approach for alleviating inflammatory bowel disease symptoms.

Dr. David Williams (Oregon State University)

We continue to utilize the mouse transplacental cancer chemoprevention model with a focus on the mechanism of transplacental polycyclic aromatic hydrocarbon (PAH)-induced cancer in the offspring as adults following in utero PAH exposure and its chemoprevention by maternal dietary supplementation with indole-3-carbinol (I3C). PAHs are formed from the incomplete combustion of organic material (coal, tobacco, wood, petroleum products, etc.). PAHs are a major health concern and represent 3 of the top 10 chemicals of concern at high priority polluted sites as evaluated by ATSDR (Agency for Toxic Substances Disease Registry, a component of the CDC). It has been estimated that 95% of exposure (for a non-smoker) to carcinogenic PAHs is through diet. We found epigenetic markers such as DNA methylation and profiles of non-coding RNAs were markedly altered in newborns from mothers exposed to a carcinogenic PAH in her diet and these alterations in the epigenome could be partially ameliorated by maternal dietary I3C.

Dr. Donato Romagnolo (University of Arizona)

The farnesoid X receptor (FXR) regulates bile acids (BA) metabolism and possesses tumor suppressor functions. The expression of FXR is reduced in colorectal tumors of subjects carrying inactivated adenomatous polyposis coli (APC). Identifying the mechanisms responsible for this reduction may offer new molecular targets for colon cancer prevention.

The objective of our current study was to investigate the influence of APC inactivation on regulation of FXR expression in colonic mucosal cells. We hypothesized that inactivation of APC epigenetically represses NR1H4 (FXR gene name) expression through increased CpG methylation. Normal proximal colonic mucosa, and normal-appearing adjacent colonic mucosa and colon tumors were collected respectively from wild-type C57BL/6J and Apc deficient (Apc^Min/+) male mice. The expression of Fxr, ileal bile acid-binding protein (Ibabp), small heterodimer partner (Shp), and cyclooxygenase-2 (Cox-2) were determined by real-time PCR. In both normal, and adjacent colonic mucosa and colon tumors, we measured CpG methylation of Fxr in bisulfonated genomic DNA. In vitro, we measured the impact of APC inactivation and deoxycholic acid (DCA) treatment on FXR expression in human colon cancer HCT-116 cells transfected with silencing RNA for APC (siAPC), and HT-29 cells carrying inactivated APC.

In Apc^{Min/ +}mice, constitutive CpG methylation of the Fxr3/4 promoter was linked to reduced (60-90%) baseline Fxr, Ibabp, and Shp, and increased Cox-2, expression in apparently normal adjacent mucosa and colon tumors. The knock-down of APC in HCT-116 cells increased c-MYC, and lowered (~50%) FXR expression, which was further reduced (~80%) by DCA. In human HCT-116, but not HT-29 colon cancer cells, DCA induced FXR expression and lowered CpG methylation of FXR. We concluded that in mouse colonic mucosa and human colon cells loss of APC function may favor silencing of FXR expression through CpG hypermethylation leading to reduced expression of downstream targets (SHP, IBABP) involved in BA homeostasis while increasing expression of factors (COX-2, c-MYC) that contribute to inflammation and colon cancer.

Future studies will examine the differential effects of various HFD (i.e. n-6 vs n-3) on FXR CpG methylation in normal and APC-deficient colon models. A second question pertains to whether other epigenetic regulatory defects associate with tumor development in APC-deficient colon cells.

OBJECTIVE 2. Elucidate mechanisms of action of dietary toxicants and develop biomarkers for human risk assessment and disease prevention.

Dr. William Helferich (University of Illinois)

A soy flour diet (MS) prevented isoflavones from stimulating MCF-7 tumor growth in athymic nude mice, indicating that other bioactive compounds in soy can negate the estrogenic properties of isoflavones. The underlying signal transduction pathways to explain the protective effects of soy flour consumption were studied. Ovariectomized athymic nude mice inoculated with MCF-7 human breast cancer cells were fed either MS or purified isoflavone mix (MI), both with equivalent amounts of genistein. Positive controls received estradiol pellets and negative controls received sham pellets. GeneChip-Human-Genome-U133-Plus-2.0 Array platform was used to evaluate gene expressions, and results were analyzed using bioinformatics approaches. Tumors in MS-fed mice exhibited higher expression of tumor-growth-suppressing genes ATP2A3 and BLNK, and lower expression of oncogene MYC. Tumors in MI-fed mice expressed higher level of oncogene MYB and lower level of MHC-I and MHC-II, allowing tumor cells to escape immunosurveillance. MS induced gene expression alterations were predictive of prolonged survival among estrogen receptor- positive breast cancer patients, whilst MI-induced gene changes were predictive of shortened survival. Our findings suggest dietary soy flour affects gene expression differently than purified isoflavones, which may explain why soy foods prevent isoflavones-induced stimulation of MCF-7 tumor growth in athymic nude mice.

Dr. David Williams (Oregon State University)

This laboratory has been the first to utilize accelerator mass spectrometry (Lawrence Livermore National Laboratory, LLNL) to follow uptake and elimination in humans of PAHs at an environmentally relevant level of exposure (10% or less of average daily exposure). Using this technology, we can measure femtograms (10^-15) of PAH/ml of blood. Ten femtograms/ml of blood is equivalent to 1 drop of water in 4000 Olympic-sized swimming pools. The current NIEHS-funded project was given an FDA IND (#117175, “Pharmacokinetics of [¹⁴C]-Benzo[a]pyrene by Micro-Dosing”) as well as Oregon State University and LLNL IRB approvals. Utilizing a new ULPC interface to the AMS instrument, we are able to measure BaP and metabolites in blood and urine rather than just [¹⁴C]. Results to date show significant metabolism (mostly BaP-7,8-dihydrodiol), along with unknown metabolites including what we believe to be conjugates with glutathione, glucuronic acid or sulfate. There is little intra-individual variation in the metabolite profile and parent BaP, but significant differences between individuals, some of which may be related to CYP1B1 and/or GSTM1 genotypes. Finally, ingestion of smoked salmon, containing a complex mixture of PAHs, along with the [¹⁴C]-BaP markedly reduces blood levels of BaP and its metabolites although we have not run a sufficient number of individuals to assess statistical significance.

Dr. Abby Benninghoff (Utah State University)

Dr. Benninghoff’s group, in collaboration with her colleagues at USU, is investigating the impacts of basal diet and dietary supplements on composition of the gut microbiome in a mouse model of colitis-associated colorectal cancer.  Mice were fed either a standard basal diet, AIN93G, or the total Western diet (TWD) for 16 weeks; at week 2 mice were dosed with azoxymethane and provided dextran sodium sulfate to induce colon tumorigenesis.  The composition of the fecal microbiome was assessed by 16S sequencing and comparisons between diet groups were made at each stage of disease development (pre-initiation, colitis, recovery, pronounced cancer).  Prior to initiation with AOM/DSS, the gut microbiome compositions of AIN93G- and TWD-fed mice were very similar.  As colitis progressed, the populations became more distinct, especially so by the recovery stage (day 45), which was also typified by sustained colonic inflammation and mucosal injury in TWD-fed mice.  By the study end (day 105), separation between the fecal microbiomes for the different diet groups was evident.  Finally, examination of the terminal fecal microbiome revealed distinct gut microbiota profiles for TWD-fed animals initiated with AOM/DSS compared to sham mice, whereas a distinction was not evident for those fed AIN93G.  In summary, mice with pronounced colitis-associated colorectal cancer, induced by dietary exposure to TWD, harbored microbiomes distinct from their counterparts fed healthy diets.

Dr. Jim Pestka (Michigan State University)

Deoxynivalenol (DON or "vomitoxin"), a trichothecene mycotoxin produced by Fusarium, is a global food safety concern because it commonly contaminates cereal grains and has the potential to cause growth suppression and gastrointestinal disease in humans. Climate change, modified agricultural practices and recent globalization of trade in agricultural plants have increased Fusarium cereal blight, thereby greatly increasing grain contamination by DON and markedly expanding the contaminant profile to include other structurally-related 8-ketotrichothecenes (3-ADON, 15-ADON, nivalenol, fusarenon X) as well as plant glucosidic metabolites such as DON-3-glucoside. Existing data are insufficient to predict the toxicity risks from exposure to mixtures in these emerging trichothecenes. We are testing the hypothesis that toxic equivalency factors (TEFs) for the 8-ketotrichothecenes derived from DON toxicity models will be applicable to food safety risk assessment and toxicity analysis. Currently we are applying this knowledge by developing a simple in vitro assay that will enable measurement of the trichothecene toxic equivalents in food samples for use as an intervention tool. Toward this end, several candidate enteroendocrine cell lines and endpoints (calcium mobilization, hormone secretion) have been identified.

OBJECTIVE 3. Discover and characterize novel bioactive dietary compounds that have beneficial or adverse effects on human health.

Dr. Michael Denison (UC Davis)

Estrogenic endocrine disrupting chemicals are found in environmental and biological samples, commercial and consumer products, food, and numerous other sources. We previously developed an estrogen-responsive recombinant human variant breast cancer cell line (vM7Luc4E2) that has been approved by the USEPA and OECD for use as a bioanalytical method to detect estrogen receptor (ER) agonists/antagonists and used this cell line to screen a wide variety of chemicals and commercial/consumer products for estrogenic activity. Although this cell line has been extremely useful for screening purposes and it is being used by many laboratories, these cells have a major limitation for comprehensive screening. vM7Luc4E2 cells contain only one of the two known ER isoforms, ERa but not ERb and the differential ligand selectivity/specificity of these two ERs indicates that the currently accepted screening method only detects a subset of the total number of estrogenic chemicals. To improve the estrogen screening bioassay, vM7Luc4E2 cells were stably transfected with an ERb expression plasmid and positive clones identified using ERb-selective ligands (genistein and Br-ERb-041). A highly responsive clone (vM7Luc4E2bc9) was identified that exhibited greater sensitivity and responsiveness to ERb-selective ligands than vM7Luc4E2 cells and qRT-PCR confirmed the presence of ERb expression in these cells. Screening of a small library of 173 pesticides and industrial chemicals not only identified chemicals that could activate both ERs but they also revealed chemicals that preferentially stimulated ERb-dependent reporter gene expression. Together, these results not only demonstrate the utility of this new dual ER recombinant cell line for detecting a broader range of estrogenic chemicals than the currently approved vM7Luc4E2 cell line, but screening with both vM7Luc4E2 and vM7Luc4E2bc9 cell lines allows identification of ERa and ERb-selective chemicals.

During the past year, we started screening commercially prepared food products for the presence of estrogenic chemicals using our recombinant cell lines. Initial screening studies revealed the presence of very high concentrations of estrogenic activity in ethanol extracts of Nabisco Nilla Wafer cookies (producing ~50% of maximal estrogenic activity with only the equivalent of 10 mg of cookie extract using the vM7Luc4E2 cell line). Given the unexpected nature of this finding, we questioned whether other cookies also contained high levels of estrogenic activity and subsequently obtained a number of different products (focusing only on cookies that lacked frosting, glazes, sprinkles or other potential confounding additives). The specific products we selected for this analysis included cookies from Nabisco (Nilla Wafers, Lorna Doones, Animal Crackers, Oreos, Saltines), Pepperidge Farms (Bordeaux, Chessman) and Walkers (Shortbread). We analyzed multiple extracts of three different lots/batches of each of these products and our studies to date revealed tremendously high estrogenic activity in vM7Luc4E2 cells incubated with extracts from the Nabisco products, with only moderate or low activity in extracts from Walkers and Pepperidge Farms products. Interestingly, substantially higher activity was detected using the new vM7Luc4E2bc9 cell line (producing up to 80% of maximal estrogenic activity with an extract volume equivalent to only 5 mg of original cookie). These results not only indicated the presence of chemicals that can activate both forms of ER, but they also revealed the presence of chemicals that selectively activated ERb signaling. Interestingly, the extracts of Lorna Doone and Animal crackers contained the highest estrogenic activity of all products examined to date (producing between 50-80% of maximal estrogenic activity with an extract that is equivalent to 5 mg of original cookie).   However, considering that the weight of one Lorna Doones is about 7 grams and a serving is 4 cookies (~28 grams), there is potential for exposure to high levels of estrogenic chemicals from consumption of one serving of cookies (if that is all an individual consumes). The presence of estrogenic chemicals in common commercial cookies has not been assessed as far as we are aware and this has opened a potentially interesting and significant area of investigation and will we continue to examine this in detail in the upcoming year. The presence of this activity is particularly of concern when we consider the potential exposure of children, who are major consumers of these products. Interestingly, the extracts showed little or no activity in the Ah receptor cell bioassays, indicating specificity of extract chemicals for the ER signaling pathway. While the extremely high levels of estrogenic activity that one could be exposed to from consumption of these cookies might turn out to be a significant concern, but this would only occur if the active chemicals can be readily absorbed and survive first-pass metabolism. These critical aspects remain to be examined in future studies.

Dr. Marie-Louise Ricketts (University of Nevada-Reno)

Dietary procyanidins are beneficial in maintaining health. Grape seed procyanidin extract (GSPE) in particular has been shown to regulate cholesterol and triglyceride (TG) homeostasis in vivo. Our lab has identified several underlying molecular mechanisms by which GSPE decreases serum TG levels, including farnesoid x receptor (FXR)-dependent decreased hepatic triglyceride synthesis, inhibition of enterohepatic bile acid (BA) recirculation and increased hepatic TG catabolism. Furthermore, we identified GSPE as a histone deacetylase (HDAC) inhibitor, leading to enhanced PPARα target-gene transcription and increased fatty acid β-oxidation. Further studies were designed to identify which component (or components) within GSPE are responsible for these effects. In an effort to facilitate further understanding regarding its molecular actions, GSPE was fractionated by column chromatography to separate compounds based on molecular size (n=3). The degree of polymerization for each fraction was determined by HPLC. Using a cell-based transient transfection assay, each fraction was tested in combination with the bile acid, chenodeoxycholic acid (CDCA), to assess its ability to increase FXR transactivation (n=3, analyzed in triplicate). Fractions that demonstrated enhanced transactivation were then analyzed by mass spectrometry. Results show that fractions enriched in dimers and dimer gallates enhance FXR transactivation. Additional in vitro studies are on going to determine whether these fractions regulate gene expression via FXR using Caco2 cells. Ultimately, this study will facilitate enhanced understanding regarding the observed GSPE-mediated regulation of metabolic homeostasis in vivo.

Dr. Mendel Friedman (USDA-ARS Albany)

Processing plant foods at elevated temperatures forms Acrylamide, a potential carcinogen. A wide range in the acrylamide concentrations has been found in commercial canned black ripe olives. This group’s work has resulted in optimized, safe thermal processing conditions that are an efficient alternative to current approaches, and leads to reduced acrylamide formation during thermal processing of olives and improved quality of black ripe olives.

Optimized techniques for analysis of food composition are needed to improve the ability to relate their function to their chemical composition. This group has worked on approaches to determine protein amino acids and metabolites, non-protein amino acids, dietary protein, glucose, fructose, sucrose, phenolic, and flavonoid content and antioxidative properties of potato tubers, peels, and pulps. The methods they describe might facilitate the analysis of potato cultivars, peels, and pulps and relate their composition and antioxidative activities to their nutritional and health-promoting properties.

Dr. Donato Romagnolo (University of Arizona)

Only 5-10% of breast cancer cases is linked to germline mutations in the BRCA-1 gene and occurs early in life. Conversely, sporadic breast tumors, which represent 90-95% of breast malignancies, have lower BRCA-1 expression, but not a mutated BRCA-1 gene, and tend to occur later in life in combination with other genetic alterations and/or environmental exposures. The latter may include environmental and dietary factors that activate the aromatic hydrocarbon receptor (AhR). Therefore, understanding if changes in expression and/or activation of the AhR are associated with somatic inactivation of the BRCA-1 gene may provide clues for breast cancer therapy.

We evaluated Brca-1 CpG promoter methylation and expression in mammary tumors induced in Sprague-Dawley rats with the AhR agonist and mammary carcinogen 7,12-dimethyl-benzo(a)anthracene (DMBA). Also, we tested in human estrogen receptor (ER)-negative sporadic UACC-3199 and ERa-positive MCF-7 breast cancer cells carrying respectively, hyper- and hypomethylated BRCA-1 gene, if the treatment with the AhR antagonist a-naphthoflavone (aNF) modulated BRCA-1 and ERa expression. Finally, we examined the association between expression of AhR and BRCA-1 promoter CpG methylation in human triple-negative (TNBC), luminal-A (LUM-A), LUM-B, and epidermal growth factor receptor-2 (HER-2)-positive breast tumor samples.

Mammary tumors induced with DMBA had reduced BRCA-1 and ERa expression; higher Brca-1 promoter CpG methylation; increased expression of Ahr and its downstream target Cyp1b1; and higher proliferation markers Ccnd1 (cyclin D1) and Cdk4. In human UACC-3199 cells, low BRCA-1 was paralleled by constitutive high AhR expression; the treatment with aNF rescued BRCA-1 and ERa, while enhancing preferential expression of CYP1A1 compared to CYP1B1. Conversely, in MCF-7 cells, aNF antagonized estradiol-dependent activation of BRCA-1 without effects on expression of ERa. TNBC exhibited increased basal AhR and BRCA-1 promoter CpG methylation compared to LUM-A, LUM-B, and HER-2-positive breast tumors.

Constitutive AhR expression coupled to BRCA-1 promoter CpG hypermethylation may be predictive markers of ERa-negative breast tumor development. Ongoing studies in our laboratory are using in vitro and vivo models to explore the effects of AhR knockout on epigenetic regulation of BRCA-1 and ESR1 (ERa, and the preventative effects of AhR antagonists. Progress in these areas may help clarifying a causative role for the AhR in breast tumorigenesis and assist with the development of risk models for BRCA-1 mutation carriers and sporadic TNBC, for which therapy options remains an intensive area of investigation.

Dr. Tiffany Weir (Colorado State University)

Noni juice is used traditionally in Polynesian cultures to relieve a variety of ailments and can be consumed fermented or non-fermented. However, fermentation alters the chemical profile and may change the bioactivity profile of the noni juice. In collaboration with Dr. Pratibha Nerurkar, who examines the effect of noni juice on high fat diet-induced diabetes in mice, we are exploring metabolomic profiles of fermented and non-fermented noni juices. We have found that fermentation is the primary source of variability in global metabolomic profiles of noni. However, these changes do not appear to be specifically associated with protection against metabolic impairments in high fat diet-fed mice.

OBJECTIVE 4. Increase beneficial or decrease adverse effects of bioactive constituents and microbes in food.

Dr. Mendel Friedman (USDA-ARS Albany)

Microbial contaminants in foods are a potential safety issue for humans and animals. Work by Friedman and colleagues have found that the glycoalkaloid tomatine found in tomatoes exhibited potent antimicrobial activity against pathogenic protozoa (Trichomonas vaginalis) that infect humans, cattle, and cats. Their results suggest its potential value for developing alternative therapy toward trichomoniasis in humans, bovines, and felines. In other studies, they found that a commercial pomegranate preparation reduced the heat resistance of the virulent pathogen Escherichia coli O104:H4 in ground chicken, suggesting that meat processors can use the described kinetic model to design lethality treatments in order to achieve specific reductions of the pathogen in ground poultry products at lower processing temperatures.

Eliminating microbial contaminants from plant-based foods is particularly problematic. This group has explored the impact of several plant-based antimicrobial washes on the sensory properties of organic leafy greens (celery, lettuce, and spinach). The showed that greens treated with 0.1% cinnamaldehyde had the highest preference liking by the 60 panelists, suggesting its value for large-scale (industrial) use to improve microbial food safety.

Dr. David Williams (Oregon State University)

A major effort this past year has been focused on reducing human health risks from food-borne pathogens and environmental contaminants. I3C and SFN are especially potent in reducing the risk to the fetus from exposure in utero to chemical carcinogens ingested in food as well as adult cancers.

Dr. Meijun Zhu (Washington State University)

Treatment of E. coli O157:H7 infection is difficult since many antibiotics are shown to induce SOS response and enhance Stx production. Thus, an alternative antimicrobial intervention of E. coli O157:H7 in human infection is needed. Cinnamon is an antimicrobial spice that has been widely used for thousands of years. We found that Cinnamon oil not only effectively inhibited growth of E. coli O157:H7 at high concentration, but also dramatically reduced both Stx2 phage and total phage induction in E. coli O157:H7 at a sub-inhibitory concentration. The inhibitory effect might be due to the suppression of bacterial SOS response key regulator RecA and vital RNA polyadenylation enzymes (PNPase and PAP I). Additionally, C. cassia oil has bactericidal effects against bovine mastitis pathogenic isolates possibly through disruption of membrane structure. These data suggest its potential application as a therapeutic to control pathogenic bacterial infections.

Amin, ARMR, Karpowicz, PA, Carey, TE, Arbiser, J, Nahta, R, Chen, ZG, Dong, J-T, Kucuk, O, Khan, GN, Huang, GS, Mi, S, Lee, H-Y, Reichrath, J, Honoki, K, Georgakilas, AG, Amedei, A, Amin, A, Helferich, B, Boosani, CS, Ciriolo, MR, Chen, S, Mohammed, SI, Azmi, AS, Keith, WN, Bhakta, D, Halicka, D, Niccolai, E, Fujii, H, Aquilano, K, Ashraf, SS, Nowsheen, S, Yang, X, Bilsland, A, Shin, DM, 2015. Evasion of anti-growth signaling: a key step in tumorigenesis and potential target for treatment and prophylaxis by natural compounds. Semin Cancer Biol. Mar 5. pii: S1044-579X(15)00013-9. doi: 10.1016/j.semcancer.2015.02.005. [Epub ahead of print] Review. PMID: 25749195

Andruska, N, Zheng, X, Yang, X, Mao, C, Cherian, MM, Mahapatra, L, Helferich, WG and Shapiro, DJ., 2015. Estrogen receptor a inhibitor activates the unfolded protein response, blocks protein synthesis and induces tumor regression. Proc Natl Acad Sci USA. Mar 30. pii: 201403685. [Epub ahead of print] PMID: 25825714

Atwell, L.L., Beaver, L.M., Shannon, J., Williams, D.E., Dashwood, R.H. and Ho, E., 2015. Epigenetic regulation by sulforaphane: Opportunities for breast and prostate cancer chemoprevention. Current Pharmacol. Reports. 1:102-111.

Atwell, L.L., Hsu, A., Beaver, L.M., Stevens, J.F., Choi, J., Jiang, Y., Bella, D., Williams, D.E., Shannon, J., Dashwood, R.H. and Ho, E., 2016. Untargeted metabolic screen reveals changes in human plasma metabolite profiles following consumption of fresh broccoli sprouts. Food Func., in press.

Aumsuwan, P, Khan, SI, Khan, IA, Ali, Z, Avula, B, Walker, LA, Shariat-Madar, Z, Helferich, WG, Katzenellenbogen, BS, Dasmahapatra, AK., 2015. The anticancer potential of steroidal saponin, dioscin, isolated from wild yam (Dioscorea villosa) root extract in invasive human breast cancer cell line MDA-MB-231 in vitro. Arch Biochem Biophys. Dec 9. pii: S0003-9861(15)30112-0. doi: 10.1016/j.abb.2015.12.001. [Epub ahead of print] PMID:26682631

Bak, S.-M., Iida, M., Soshilov, A.A., Denison, M.S., Iwata, H. and Kim, E.-Y., 2016. Auto-induction mechanism of aryl hydrocarbon receptor 2 (AHR2) gene by TCDD-activated AHR1 and AHR2 in the red seabream (Pagrus major), Arch. Toxicol., In Press. PMID: 27188387

Banerjee, N., Kim, H., Talcott, S.T., Turner, N.D., Byrne, D.H., and Mertens-Talcott, S.U., 2016. Plum polyphenols inhibit colorectal aberrant crypt foci formation in rats: Potential role of the miR-143/protein kinase B/mammalian target of rapamycin axis. Nutrition Research 36, 1105-1113.

Bates, M.A., Brandenberger, C., Langohr, II, Kumagai, K., Lock, A.L., Harkema, J.R., Holian, A., Pestka, J.J., 2016. Silica-triggered autoimmunity in lupus-prone mice blocked by docosahexaenoic acid consumption. PLoS One 11, e0160622.

Bibi, S., Kang, Y., Yang, G., and Zhu, M.J., 2016. Grape seed extract improves small intestinal morphology through suppressing inflammation and regulating alkaline phosphatase in IL10-deficient mice. Journal of Functional Food, 20: 245-252.

Block, KI, Gyllenhaal, C, Lowe, L, Amedei, A, Amin, ARMR, et al., 2015. A broad-spectrum integrative design for cancer prevention and therapy. Semin Cancer Biol. 35 Suppl:S276-304. doi: 10.1016/j.semcancer.2015.09.007. PMID:26590477

Boonmuen, N, Gong, P, Ali, Z, Chittiboyina, AG, Khan, I, Doerge, DR, Helferich, WG, Carlson, KE, Martin, T, Piyachaturawat, P, Katzenellenbogen, JA, Katzenellenbogen, BS., 2015. Licorice root components in dietary supplements are selective estrogen receptor modulators with a spectrum of estrogenic and anti-estrogenic activities. Steroids. 105, 42-49. doi: 10.1016/j.steroids.2015.11.006. [Epub ahead of print] PMID:26631549

Borresen, E., Brown, D.G., Harbison, G., Taylor, L., Fairbanks, A., O’Malia, J., Bazan, M., Rao, S., Baily, S., Wdowik, M., Weir, T.L., Brown, R.J., Ryan, E.P., 2016. A randomized-controlled trial to increase navy bean or rice bran consumption in colorectal cancer survivors. Nutrition & Cancer. Dx.doi.org/10.1080/01635581.2016.1224370.

Brennan, J.C., Bassal, A., He, G. and Denison, M.S., 2016. Development of a recombinant human ovarian (BG1) cell line containing estrogen receptor a and b for improved detection of estrogenic/antiestrogenic chemicals. Environ. Toxicol. Chem. 35, 91-100. PMC4772679

Brown, D.G., Rao, S., Weir, T.L., O’Malia, J., Bazan, M., Brown, R.J., Ryan, E.P., 2016. Metabolomics investigation of tumors, adjacent colonic mucosa and stool from colorectal cancer patients. Cancer & Metabolism 4:11.

Cai, K, Yen, J, Yin, Q, Liu, Y, Song, Z, Lezmi, S, Zhang, Y, Yang, X, Helferich, WG, and Cheng, J., 2015. Redox-responsive self-assembled chain-shattering polymeric therapeutics. Biomater Sci. 3, 1061-1065. PMID: 26146551

Casey, SC, Amedei, A, Aquilano, K, Benencia, F, Bhakta, D, Boosani, CS, Chen, S, Ciriolo, MR, Crawford, S, Fujii, H, Georgakilas, AG, Guha, G, Halicka, D, Helferich, WG, Heneberg, P, Honoki, K, Kerkar, SP, Mohammed, SI, Niccolai, E, Nowsheen, S, Vasantha Rupasinghe, HP, Samadi, A, Singh, N, Talib, WH, Venkateswaran, V, Whelan, RL, Yang, X, Felsher, DW., 2015. Cancer prevention and therapy through the modulation of the tumor microenvironment. Semin Cancer Biol. Apr 9. pii: S1044-579X(15)00015-2. doi: 10.1016/j.semcancer.2015.02.007. [Epub ahead of print] Review. PMID: 25865775.

Choi, S.-H., Kozukue, N., and Friedman, M., 2016. Composition and antioxidative and cancer cell inhibiting activities of Jujube fruits and seeds (Ziziphus jujuba) cultivated in Korea. In Chinese Dates: A Traditional Functional Food, Liu, D.; Ye, X.; Jiang, Y., Eds. CRC Press: Boca Raton, FL, pp 99-114.

Choi, S.-H., Kozukue, N., Kim, H.-J., and Friedman, M., 2016. Analysis of protein amino acids, non-protein amino acids and metabolites, dietary protein, glucose, fructose, sucrose, phenolic, and flavonoid content and antioxidative properties of potato tubers, peels, and cortexes (pulps). J. Food Compos. Anal. 50, 77-87.

Clark, E.S., Flannery, B.M., Gardner, E.M., Pestka, J.J., 2015a. High sensitivity of aged mice to deoxynivalenol (Vomitoxin)-induced anorexia corresponds to elevated proinflammatory cytokine and satiety hormone responses. Toxins (Basel) 7, 4199-4215.

Clark, E.S., Flannery, B.M., Pestka, J.J., 2015b. Murine anorectic response to deoxynivalenol (vomitoxin) is sex-dependent. toxins (Basel) 7, 2845-2859.

Cole, B., Brander, S., Jeffries, K., Hasenbein, S., He, G., Denison, M.S., Fangue, N. and Connon, R., 2016. Changes in menidia beryllina gene expression and in vitro hormone receptor activation following exposure to estuarine waters near treated wastewater outfalls, Arch. Environ. Contam. Toxicol. 71, 210-223. PMID: 27155869

Corrada, D., Soshilov, A.A., Denison, M.S. and Bonati, L., 2016. Deciphering dimerization modes of PAS domains: Computational and experimental analyses of the AhR:ARNT complex reveal new insights into the mechanisms of AhR transformation, PLoS Comp. Biol. 12(6), e1004981. PMC4905635

Cox-York, K, Sheflin, AM, Foster, MT, Gentile, CL, Kahl, A, Koch, L, Britton, S, Weir, TL, 2015. Gut ecology associated with high innate aerobic capacity protects against ovariectomy-induced cardiometabolic risk. Physiol Reports 3, e12488.

Croes, K., Van den Heuvel, R., Van den Bril, B., Staelens, J., Denison, M.S., Van Langenhove, K., Vandermarken, T. and Elskens, M., 2016. Assessment of estrogenic and androgenic activity in PM10 air samples from an urban, industrial and rural area in Flanders (Belgium) using the CALUX bioassay. Environ Res. 150, 66-72. PMID: 27257826

Doerge, DR, Woodling, KA, Churchwell, MI, Fleck, SC, and Helferich, WG., 2016. Pharmacokinetics of isoflavones from soy infant formula in neonatal and adult rhesus monkeys. Food Chem Toxicol. Apr 12. pii: S0278-6915(16)30111-9. doi: 10.1016/j.fct.2016.04.005. [Epub ahead of print] PMID:27084109

Donovan, M.G., Selmin, O.I., Doetschman, T.C., Romagnolo, D.F. 2016. Mediterranean diet, inflammatory bowel diseases and colon cancer. In: Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer Pubs, Eds. Romagnolo, Selmin. Chapter 14, pg. 181-201.

Downing, L.E., Ferguson, B.S., Rodriguez, K., Ricketts, M.L., 2016. A grape seed procyanidin extract inhibits HDAC activity leading to increased Pparα phosphorylation and target-gene expression. Mol Nutr Food Res. Sep 14. doi: 10.1002/mnfr.201600347. [Epub ahead of print] PMID: 27624175

Downing, L.E., Heidker, R.M., Caiozzi, G.C., Wong, B.S., Rodriguez, K., Del Rey, F., Ricketts, M.L., 2015. A grape seed procyanidin extract ameliorates fructose-induced hypertriglyceridemia in rats via enhanced fecal bile acid and cholesterol excretion and inhibition of hepatic lipogenesis. PLoS One. 10(10):e0140267. doi: 10.1371/journal.pone.0140267. PMID: 26458107

Ferguson, LR, Chen, H, Collins, AR, Connell, M, Damia, G, Dasgupta, S, Malhotra, M, Meeker, AK, Amedei, A, Amin, A, Ashraf, SS, Aquilano, K, Azmi, AS, Bhakta, D, Bilsland, A, Boosani, CS, Chen, S, Ciriolo, MR, Fujii, H, Guha, G, Halicka, D, Helferich, WG, Keith, WN, Mohammed, SI, Niccolai, E, Yang, X, Honoki, K, Parslow, VR, Prakash, S, Rezazadeh, S, Shackelford, RE, Sidransky, D, Tran, PT, Yang, ES, Maxwell, CA., 2015. Genomic instability in human cancer: Molecular insights and opportunities for therapeutic attack and prevention through diet and nutrition. Semin Cancer Biol. Apr 10. pii: S1044-579X(15)00020-6. doi: 10.1016/j.semcancer.2015.03.005. [Epub ahead of print] Review. PMID: 25869442.

Foster, M.T., Gentile, C.L., Cox-York, K., Wei, Y., Wang, D., Estrada, A., Reese, L., Miller, T., Pagliassotti, M.J., Weir, T.L., 2016. Fuzhuan tea consumption imparts hepatoprotective effects and alters intestinal microbiota in high saturated fat diet-fed rats. Molecular Nutrition and Food Research 60, 1213-1220.

Friedman, M., 2016. Bioactive compounds from Ziziphus jujuba and allied species. In Chinese Dates: A Traditional Functional Food, Liu, D.; Ye, X.; Jiang, Y., Eds. CRC Press: Boca Raton, FL, pp 35-52.

Friedman, M., and Levin, C.E., 2016. Glycoalkaloids and calystegine alkaloids in potatoes. In Advances in Potato Chemistry and Technology, 2nd ed.; Singh, J.; Kaur, L., Eds. Elsevier: Oxford, UK, pp 167-194.

Friedman, M., Levin, C.E., and Henika, P.R., 2016. Addition of phytochemical-rich plant extracts mitigate the antimicrobial activity of essential oil/wine mixtures against Escherichia coli O157:H7 but not against Salmonella enterica. Food Control Online Sept 4, doi: 10.1016/j.foodcont.2016.09.002.

Fu, X., Zhu, M.J., Zhang, S., Foretz, M., Viollet, B., and Du, M., 2016. Obesity impairs skeletal muscle regeneration via inhibition of AMP-activated protein kinase. Diabetes, 65:188-200.

Harper, T.A., Jr., Morré, J., Lauer, F.T., McQuistan, T.J., Hummel, J.M., Burchiel, S.W. and Williams, D.E., 2015. Analysis of dibenzo[def,p]chrysene-deoxyadenosine adducts in wild-type and cytochrome P450 1b1 knockout mice using stable-isotope dilution UHPLC-MS/MS. Mutat. Res. Gen. Toxicol. Environm. Mutag. 782:51-56.

Heidker, R.M., Caiozzi, G.C., Ricketts, M.L., 2016. Dietary procyanidins selectively modulate intestinal farnesoid X receptor-regulated gene expression to alter enterohepatic bile acid recirculation: elucidation of a novel mechanism to reduce triglyceridemia. Mol Nutr Food Res. 60, 727-36. doi: 10.1002/mnfr.201500795. PMID: 26718753

Heidker, R.M., Caiozzi, G.C., Ricketts, M.L., 2016. Grape seed procyanidins and cholestyramine differentially alter bile acid and cholesterol homeostatic gene expression in mouse intestine and liver. PLoS One. 11(4):e0154305. doi: 10.1371/journal.pone.0154305. PMID: 27111442

Hong, M.Y., Turner, N.D., Murphy, M.E., Carroll, R.J., Chapkin, R.S., and Lupton, J.R., 2015. In vivo regulation of colon cell proliferation, differentiation, apoptosis and P27Kip1 by dietary fish oil and butyrate in rats. Cancer Prevention Research 8, 1076-1083. PMID 26323483.

Hwang, P., Oishi, M., Karratti-Abordo, J., and Nerurkar, P.V., Fermented noni juice induces apoptosis by differentially regulating survivin signaling in ER positive, MCF-7 and triple negative, MDA-MB-231 breast cancer cells. Oncotarget, Submitted, 2016

Jiang, WG, Sanders, AJ, Katoh, M, Ungefroren, H, Gieseler, F, Prince, M, Thompson, SK, Zollo, M, Spano, D, Dhawan, P, Silva, D, Subbarayan, PR, Sarkar, M, Honoki, K, Fujii, H, Georgakilas, AG, Amedei, A, Niccolai, E, Amin, A, Ashraf, SS, Ye, L, Helferich, WG, Yang, X, Boosani, CS, Guha, G, Ciriolo, MR, Aquilano, K, Chen, S, Azmi, AS, Keith, WN, Bilsland, A, Bhakta, D, Halicka, D, Nowsheen, S, Pantano, F, Santini, D., 2015. Tissue invasion and metastasis: Molecular, biological and clinical perspectives. Semin Cancer Biol. Apr 9. pii: S1044-579X(15)00023-1. doi: 10.1016/j.semcancer.2015.03.008. [Epub ahead of print] Review. PMID: 25865774.

Johnson, G.S., Li, J., Beaver, L., Dashwood, W.-M., Sun, D., Rajendran, P., Williams, D.E., Ho, E. and Dashwood, R.H., 2016. A functional pseudogene, NMRAL2P, is regulated by Nrf2 and serves as a co-activator of NQO1 in sulforaphane-treated colon cancer cells. Molec. Nutr. Food Res., in press.

Johnson, N.B., Deck, K.M., Nizzi, C.P. and Eisenstein, R.S. A synergistic role of IRP1 and FBXL5 in controlling iron metabolism during cell proliferation. To be submitted 11/16.

Joshi, K., Parks, P., Ravishankar, S., and Friedman, M., 2016. The impact of plant-based antimicrobials on sensory properties of organic leafy greens. Food Nutr. Sci. 7, 906-919.

Juneja, V.K., Cadavez, V., Gonzales-Barron, U., Mukhopadhyay, S. and Friedman, M., 2016. Effect of pomegranate powder on the heat inactivation of Escherichia coli O104:H4 in ground chicken. Food Control 70, 26-34.

Kang, Y., Xue, Y., Du, M., Zhu, M.J., 2016. Preventative effects of Goji berry in dextran sulfate sodium-induced colitis. Journal of Nutritional Biochemistry, accepted.

Kim, E., Davidson, L.A., Zoh, R.S., Patil, B.S., Jayaprakasha, G.K., Callaway, E.S., Allred, C.D., Turner, N.D., and Chapkin, R.S., 2016. Homeostatic responses of colonic LGR5 stem cells following acute in vivo exposure to a genotoxic carcinogen. Carcinogenesis 37, 206-214.

Kim, E., Davidson, L.A., Zoh, R.S., Hensel, M.E., Salinas, M.L., Patil, B.S., Jayaprakasha, G.K., Callaway, E.S., Allred, C.D., Turner, N.D., Weeks, B., and Chapkin, R.S., 2016. Rapidly cycling Lgr5+ stem cells are exquisitely sensitive to extrinsic dietary factors that modulate colon cancer risk. Cell Death & Disease 7, e2460: doi:10.1038/cddis.2016.269.

Kim, S.-P., Lee, S.-J., Nam, S.-H., and Friedman, M., 2016. Elm tree (Ulmus parvifolia) bark bioprocessed with mycelia of Shiitake (Lentinus edodes) mushrooms in liquid culture: composition and mechanism of protection against allergic asthma in mice. J. Agric. Food Chem. 64, 773-84.

Kunde, D., Chong, W., Nerurkar, P., Ahuja, K., Just, J., Smith, J., Guven, N., and Eri, R. Bitter melon protects against ER stress in LS174T colonic epithelial cells. BMC Complementary and Alternative Medicine. Under revision, 2016

Lee, D.M., Battson, M.L., Jarrell, D., Cox-York, K., Foster, M.T., Weir, T.L., Gentile, C.L., 2016. Fuzhuan tea reverses arterial stiffening following modest weight gain in mice. Nutrition. Dx.doi.org/10.1016/j.nut.2016.07.010

Lemas, D.J., Young, B.E., Baker, P.R., Tomczik, A., Soderborg, T.K., Hernandez, T.L., de la Houssaye, B.A., Robertson, C.E., Rudolph, M.C., Ir, D., Patinkin, Z.W., Krebs, N.F., Santorico, S.A., Weir, T.L., Barbour, L.A., Frank, D.N., Friedman, J.E., 2016. Alterations in human milk leptin and insulin impact early changes in the infant gut microbiome. American Journal Clinical Nutrition. 103, 1291.

Liang, X., Yang, Q., Fu, X., Rogers, C.J., Wang, B., Pan, H., Zhu, M.J., Nathanielsz, P.W., and Du, M., 2016. Maternal obesity epigenetically alters visceral fat progenitor cell properties in male offspring mice. Journal of Physiology, 594: 4453-666

Liu, Y., Hilakivi-Clarke, L., Zhang, Y., Wang, X., Pan, YX., Xuan, J., Fleck, S.C., Doerge, D.R., Helferich, W.G., 2015. Isoflavones in soy flour diet have different effects on whole-genome expression patterns than purified isoflavone mix in human MCF-7 breast tumors in ovariectomized athymic nude mice. Mol Nutr Food Res. Mar 27. doi: 10.1002/mnfr.201500028. [Epub ahead of print] PMID:25820259.

Luo, T., Snyder, S.M., Zhao, B., Sullivan, D.K., Hamilton-Reeves, J., Guthrie, G., Ricketts, M.L., Shiverick, K.T., Shay, N., 2016. Gene expression patterns are altered in athymic mice and metabolic syndrome factors are reduced in c57bl/6j mice fed high-fat diets supplemented with soy isoflavones. J Agric Food Chem. 64, 7492-7501. PMID: 27653593

Madak, Erdogan Z, Gong, P, Zhao, YC, Xu, L, Wrobel, KU, Hartman, JA, Wang, M, Cam, A, Iwaniec, UT, Turner, RT, Twaddle, NC, Doerge, DR, Khan, IA, Katzenellenbogen, JA, Katzenellenbogen, BS, and Helferich, WG., 2015. Dietary licorice root supplementation improves diet-induced weight gain, lipid deposition and hepatic steatosis in ovariectomized mice without stimulating reproductive tissues and mammary gland. Mol Nutr Food Res. Nov 10. doi: 10.1002/mnfr.201500445. [Epub ahead of print]. PMID:26555669

Madeen, E.P., Löhr, C.V., You, H., Siddens, L.K., Krueger, S.K., Dashwood, R.H., Gonzalez, F.J., Baird, W.M., Ho, E., Bramer, L., Waters, K.M. and Williams, D.E., 2016. Dibenzo[def,p]chrysene transplacental exposure in wild-type, Cyp1b1 knockout, and CYP1B1 humanized mice. Molec. Carcinogenesis, March 17, DOI 10.1002/mc.22480 [Epub ahead of print].

Madeen, E.P., Ognibene, T.J., Corley, R.A., McQuistan, T.J., Baird, W.M., Bench, G., Turteltaub, K.W. and Williams, D.E., 2016. Human micro-dosing with carcinogenic polycyclic aromatic hydrocarbons: In vivo pharmacokinetics of dibenzo[def,p]chrysene and metabolites by UPLC accelerator mass spectrometry. Chem. Res. Toxicol., in press.

Madeen, E.P. and Williams, D.E. 2016. Environmental PAH exposure and male idiopathic infertility: A review on early life exposures and adult diagnosis. Rev. Environ. Hlth., in press.

Mahalingam, S, Gao, L, Gonnering, M, Helferich, WG, Flaws, JA., 2016. Equol inhibits growth, induces atresia, and inhibits steroidogenesis of mouse antral follicles in vitro. Toxicol Appl Pharmacol. 295, 47-55. doi: 10.1016/j.taap.2016.02.009. [Epub ahead of print] PMID:26876617

Male, D., Wu, W., Mitchell, N.J., Bursian, S., Pestka, J.J., Wu, F., 2016. Modeling the emetic potencies of food-borne trichothecenes by benchmark dose methodology. Food Chem Toxicol 94, 178-185.

Mexia, N., Gaitanis, G., Velegraki, A., Soshilov, A., Denison, M.S. and Magiatis, P., 2015. Pityriazepin and other potent AhR ligands isolated from Malassezia furfur yeast, Arch. Biochem. Biophys. 571, 16-20. PMC4454357

Mohammad, RM, Muqbil, I, Lowe, L, Yedjou, C, Hsu, HY, Lin, LT, Siegelin, MD, Fimognari, C, Kumar, NB, Dou, QP, Yang, H, Samadi, AK, Russo, GL, Spagnuolo, C, Ray, SK, Chakrabarty, M, Morre, JD, Coley, HM, Honoki, K, Fujii, H, Georgakilas, AG, Amedei, A, Niccolai, E, Amin, A, Ashraf, SS, Helferich, WG, Yang, X, Boosani, CS., Guha, G, Bhakta, D, Ciriolo, MR, Aquilano, K, Chen, S, Mohammed, SI, Keith, WN, Bilsland, A, Halicka, D, Nowsheen, S, Azmi, AS., 2015. Broad targeting of resistance to apoptosis in cancer. 2015 Semin Cancer Biol. Apr 28. pii: S1044-579X(15)00016-4. DOI: 10.1016/j.semcancer.2015.03.001. [Epub ahead of print]. PMID: 25936818

Monsanto, S.P., Hintze, K.J., Ward, R.E., Larson, D.P., Lefevre, M., Benninghoff, A., 2016. The new total Western diet for rodents does not induce an overweight phenotype or alter parameters of metabolic syndrome in mice. Nutrition Research, 36, 1031-1044. dx.doi.org/10.1016/j.nutres.2016.06.002

Monson, M.S., Cardona, C.J., Coulombe, R.A., and Reed, K.M., 2016. Hepatic transcriptome responses of domesticated wild turkey embryos to Aflatoxin B1. Toxins 8 (1). doi: 10.3390/toxins8010016

Monson, M. S., Coulombe, R.A., and Reed, K.M., 2016. Sensitivity and response of poultry to aflatoxin B1 exposure. Agriculture doi:10.3390/agriculture50x000x.

Monson, M. S., Settlage, R.E., Mendoza, K.M., Rawal, S., El-Nezami, H., Coulombe, R.A., and K.M. Reed, K.M., 2015. Modulation of the spleen transcriptome in domestic turkey (Meleagris gallopavo) in response to aflatoxin B1 and probiotics. Immunogenetics DOI 10.1007/s00251-014-0825-y

Nerurkar, P.V., Hwang, P.W., Saksa, E., 2015. Anti-diabetic potential of noni: The yin and the yang. Molecules. 20, 17684-719. doi: 10.3390/molecules201017684.

Patel, S, Peretz, J, Pan, YX, Helferich, WG, Flaws, JA., 2016. Genistein exposure inhibits growth and alters steroidogenesis in adult mouse antral follicles. Toxicol Appl Pharmacol. 293, 53-62. doi: 10.1016/j.taap.2015.12.026. Epub 2016 Jan 12. PMID:26792615

Rada, X., Todd, J., Friedman, M., Patel, J., Jaroni, D., and Ravishankar, S., 2016. Combining essential oils and olive extract for control of multi-drug resistant Salmonella enterica on organic leafy greens. SDRP Journal of Food Science & Technology 1, 1-9.

Rendeiro, C, Sheriff, A, Bhattacharya, TK, Gogola, JV, Baxter, JH, Chen, H, Helferich, WG, Roy, EJ, Rhodes, JS., 2016. Long-lasting impairments in adult neurogenesis, spatial learning and memory from a standard chemotherapy regimen used to treat breast cancer. Behav Brain Res. 315, 10-22. doi: 10.1016/j.bbr.2016.07.043. [Epub ahead of print] PMID:27478140

Rensvold, J.W., Krautkramer, K.A., Dowell, J.A., Denu, J.M., and Pagliarini, D.J., 2016. Iron deprivation induces transcriptional regulation of mitochondrial biogenesis. Journal of Biological Chemistry, 291, 20827-20837

Romagnolo, D.F., Jackson, K.A., Sparks, P.L., Selmin, O.I. 2016. Building the Mediterranean Pyramid-PART B:  Balancing the plate. In: Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer, Eds. Romagnolo, Selmin. Chapter 20, pg. 275-288.

Romagnolo, D.F., Papoutsis, A.J., Laukaitis, C., Selmin, O.I. 2015. Constitutive expression of AhR and BRCA-1 promoter CpG hypermethylation as biomarkers of ERα-negative breast tumorigenesis. BMC Cancer. 15, 1026.

Romagnolo, D.F., Selmin, O.I. Co-Editors. 2016. Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer.

Romagnolo, D.F., Selmin, O.I. 2015. Epigenetics of Endocrine Tumors in Women and Dietary Prevention. In: Preventive Nutrition, Fifth Edition, Humana Press/Springer, Eds. Bendich, Deckelbaum, Chapter 9, pg. 153-166.

Romagnolo, D.F., Selmin, O.I. 2016. Mediterranean diet and lifestyle in a modern world context In: Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer, Eds. Romagnolo, Selmin. Chapter 2, pg. 15-26.

Schreurs, A-S., Shirazi-Fard, Y., Shahnazari, M., Alwood, J.S., Truong, T.A., Tahimic, C.G.T., Limoli, C.L., Turner, N.D., Halloran, B., and Globus, R.K., 2016. Dried plum diet protects from bone loss caused by ionizing radiation. Scientific Reports DOI: 10.1038/srep21343. PMID 26867002.

Selmin, O.I., Daniels, K.D., Grunwald, J.T., Ramos, S.A., Propper, C.R., Romagnolo, D.F. 2016. Epigenetics of breast cancer: modifying role of environmental and bioactive food compounds. Mol Nutr Food Res. 60, 1310-29.

Selmin, O.I., Fang, C., Lyon, A.M., Doetschman, T.C., Thompson, P.A., Martinez, J.D., Smith, J.W., Lance, P.M., Romagnolo, D.F. 2016. Inactivation of adenomatous polyposis coli reduces bile acid/farnesoid X receptor expression through Fxr gene CpG methylation in mouse colon tumors and human colon cancer cells. J Nutr. 146, 236-4.

Selmin, O.I., Romagnolo, A.P.G., Romagnolo, D.F. 2016. The Mediterranean diet and neurodegenerative diseases. In: Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer, Eds. Romagnolo, Selmin. Chapter 12, pg. 153-164.

Selmin, O.I., Romagnolo, D.F. 2016. Building the Mediterranean Pyramid-PART A: Mediterranean recipes. In: Mediterranean Diet: Dietary Guidelines and Impact on Health and Disease. Humana Press/Springer, Eds. Romagnolo, Selmin. Chapter 19, pg. 261-273.

Sheflin, A.M., Borresen, E.C., Kirkwood, J., Boot, C., Whitney, A.K., Lu, S., Brown, R.J., Broeckling, C.D., Ryan, E.P., Weir, T.L., 2016. Dietary supplementation with rice bran or navy bean alters gut bacterial metabolism in colorectal cancer survivors. Molecular Nutrition and Food Research. 10.1002/mnfr.201500905.

Sheng, L., Rasco, B., and Zhu, M.J., 2016. Cinnamon oil inhibited Shiga toxin 2 phage induction and Shiga toxin 2 production in E. coli O157:H7. Applied Environmental Microbiology, 82, in press.

Sheng, L., Olsen, S.A., Hu, J., Yue, W., Means, W.J., and Zhu, M.J., 2016. Grape seed extract regulates virulence factors of CDC “top-six” non-O157 STEC through inhibiting quorum sensing. International Journal of Food Microbiology, 229: 24-32.

Strakovsky, RS, Lezmi, S, Shkoda, I, Flaws, JA, Helferich, WG, Pan, YX., 2015. In utero growth restriction and catch-up adipogenesis after developmental di (2-ethylhexyl) phthalate exposure cause glucose intolerance in adult male rats following a high-fat dietary challenge. J Nutr Biochem. Jun 20. pii: S0955-2863(15)00145-X. doi: 10.1016/j.jnutbio.2015.05.012. [Epub ahead of print]. PMID:26188368

Strakovsky, RS, Wang, H, Engeseth, NJ, Flaws, JA, Helferich, WG, Pan, YX, Lezmi, S., 2015. Developmental bisphenol A (BPA) exposure leads to sex-specific modification of hepatic gene expression and epigenome at birth that may exacerbate high-fat diet-induced hepatic steatosis. Toxicol Appl Pharmacol. 284, 101-12. PMID: 25748669

Tang, S., Avena-Bustillos, R.J., Lear, M., Sedej, I., Holstege, D.M., Friedman, M., McHugh, T.H., Wang, S.C., 2016. Evaluation of thermal processing variables for reducing acrylamide in canned black ripe olives. J. Food Eng. 191, 124-130.

Teeguarden, J.G., Tan, Y.-M., Edwards, S.W., Leonard, J.A., Anderson, K.A., Corley, R.A., Harding, A.K., Kile, M.L., Simonich, S.M., Stone, D., Tanguay, R.L., Waters, K.M., Harper, S.L. and Williams, D.E., 2016. Completing the link between exposure science and toxicology for improved environmental health decision making: The aggregate exposure pathway framework. Environ. Sci. Technol., Feb. 10, Epub ahead of print, PMID: 26759916. *Featured manuscript of this ES&T issue.

Vasta, J.D., Andersen, K.A., Deck, K.M., Nizzi, C.P., Eisenstein, R.S. and Raines, R.T., 2016. Selective inhibition of collagen prolyl-4-hydroxylase in human cells. ACS Chemical Biology 11:193-9

Vinay, DS, Ryan, EP, Pawelec, G, Talib, WH, Stagg, J, Elkord, E, Lichtor, T, Decker, WK, Whelan, RL, Hmc, SK, Signori, E, Honoki, K, Georgakilas, AG, Amin, A, Helferich, WG, Boosani, CS, Guha, G, Ciriolo, MR, Chen, S, Mohammed, SI, Azmi, AS, Keith, WN, Bhakta, D, Halicka, D, Fujii, H, Aquilano, K, Ashraf, SS, Nowsheen, S, Yang, X, Choi, BK, Kwon, BS., 2015. Immune evasion in cancer: Mechanistic basis and therapeutic strategies. Seminars in Cancer Biology Mar 24. pii: S1044-579X(15)00019-X. doi: 10.1016/j.semcancer.2015.03.004. [Epub ahead of print] PMID: 25818339

Wang, B., Yang, Q., Harris, C.L., Nelson, M.L., Busboom, J.R., Zhu, M.J., and Du, M., 2016. Nutrigenomic regulation of adipose tissue development – role of retinoic acid. Meat Science, 120: 100-106.

Wang, R., Kang, Y., Löhr, C.V., Fischer, K.A., Bradford, S.S., Johnson, G., Dashwood, W.-M., Williams, D.E., Ho, E. and Dashwood, R.H., 2016. Reciprocal regulation of BMF and BIRC5 (survivin) linked to Eomes overexpression in colorectal cancer. Cancer Lett. 381, 341-348. PMID:27539959

Wang, W, Belosay, A, Yang, X, Hartman, JA, Song, H, Iwaniec, UT, Turner, RT, Churchwell, MI, Doerge, DR, and Helferich, WG., 2016. Effects of letrozole on breast cancer micro-metastatic tumor growth in bone and lung in mice inoculated with murine 4T1 cells. Clin Exper Metas, 33, 475-85. doi: 10.1007/s10585-016-9792-z. Epub 2016 May 21. PMID:27209469

Wang, Z, Dabrosin, C, Yin, X, Fuster, MM, Arreola, A, Rathmell, WK, Generali, D, Nagaraju, GP, El-Rayes, B, Ribatti, D, Chen, YC, Honoki, K, Fujii, H, Georgakilas, AG, Nowsheen, S, Amedei, A, Niccolai, E, Amin, A, Ashraf, SS, Helferich, B, Yang, X, Guhas, G, Bhaktas, D, Ciriolot, MR, Aquilanot, K, Chen, S, Halicka, D, Mohammed, SI, Azmi, AS, Bilsland, A, Keith, WN, Jensen, LD., 2015. Broad targeting of angiogenesis for cancer prevention and therapy. Semin Cancer Biol. Jan 16. pii:S1044-579X(15)00002-4. doi: 10.1016/j.semcancer.2015.01.001. [Epub ahead of print] Review. PMID: 25600295

Ward, R.E., Benninghoff, A., Healy, B.J., Li, M., Vagu, B., Hintze, K.J., Consumption of the total Western diet differentially affects the response to green tea in rodent models of chronic disease compared to the AIN93G diet. Molecular Nutrition and Food Research, (in press).

Watson, G.W., Wickramasekara, S., Maier, C.S., Williams, D.E., Dashwood, R.H. and Ho, E., 2015. Assessment of global proteome in LNCaP cells by 2D-RP/RP LC-MS/MS following sulforaphane exposure. EuPA Open Proteomics Aug. 15, 2015, DOI 10.1016/j.euprot.2015.08.002.

Watson, G.W., Wickramasekara, S., Fang, Y., Maier, C.S., Williams, D.E., Dashwood, R.H., Perez, V.I. and Ho, E., 2015. HDAC activity is not required for basal autophagic flux in metastatic prostate cancer cells. Exp. Biol. Med., Dec. 6. Pii: 1535370215618518.

Whitney, A.K. and Weir, T.L., 2016. Inhibitory activity of Fuzhuan tea fractions against Salmonella enterica and enteric pathogens. Current Topics in Phytochemistry 13, 17-27.

Whitney, A.K. and Weir, T.L., 2015. Interaction of caffeine with the SOS response pathway in Escherichia coli. Gut Pathogens 7, 21.

Wu, W., Zhou, H.R., Bursian, S.J., Link, J.E., Pestka, J.J., 2016a. Calcium-sensing receptor and transient receptor ankyrin-1 mediate emesis induction by deoxynivalenol (vomitoxin). Toxicol Sci.

Wu, W., Zhou, H.R., Bursian, S.J., Link, J.E., Pestka, J.J., 2016b. Emetic responses to T-2 toxin, HT-2 toxin and emetine correspond to plasma elevations of peptide YY3-36 and 5-hydroxytryptamine. Arch Toxicol 90, 997-1007.

Wu, W., Zhou, H.R., Pestka, J.J., 2016c. Potential roles for calcium-sensing receptor (CaSR) and transient receptor potential ankyrin-1 (TRPA1) in murine anorectic response to deoxynivalenol (vomitoxin). Arch Toxicol.

Xue, Y., Zhang, H., Sun, X., and Zhu, M.J., 2016. Metformin improves ileal epithelial barrier function in interleukin-10 deficient mice. PLoS ONE, accepted.

Yang, G., Bibi, S., Du, M., Suzuki, T., and Zhu, M.J., 2016. Regulation of the intestinal tight junction by natural polyphenols: a mechanistic perspective. Critical Reviews in Food Science and Nutrition, in press.

Yang, Q., Liang, X., Sun, X., Fu, X., Zhang, L., Rogers, C.J., Wang, S., Wang, B., Maricelli, J.W., Viollet, B., Rodger, B.D., Zhu, M.J., and Du, M., 2016. AMPK/α-ketoglutarate axis dynamically mediates DNA demethylation in the Prdm16 promoter and brown adipogenesis. Cell Metabolism, in press.

Yang, X., Belosay A., Hartman, J.A., Song H., Zhang Y., Wang W., Doerge D.R. and Helferich W.G., 2015. Dietary soy isoflavones increase metastasis to lungs in an experimental model of breast cancer with bone micro-tumors. Clin Exp Metastasis. 32, 323-333. PMID: 25749878

Yaswen, P, MacKenzie, KL, Keith, WN, Hentosh, P, Rodier, F, Zhu, J, Firestone, GL, Matheu, A, Carnero, A, Bilsland, A, Sundin, T, Honoki, K, Fujii, H, Georgakilas, AG, Amedei, A, Amin, A, Helferich, B, Boosani, CS, Guha, G, Ciriolo MR, Chen, S, Mohammed, SI, Azmi, AS, Bhakta, D, Halicka, D, Niccolai, E, Aquilano, K, Ashraf, SS, Nowsheen, S, Yang, X., 2015. Therapeutic targeting of replicative immortality. Semin Cancer Biol. Apr 10. pii: S1044-579X(15)00022-X. doi: 10.1016/j.semcancer.2015.03.007. [Epub ahead of print] Review. PMID: 25869441

Yin, Q., Tang, L., Cai, K., Tong, R., Sternberg, R., Yang, X., Dobrucki, L.W., Borst, L.B., Kamstock, D., Song, Z., Helferich, W.G., Cheng, J., Fan, T.M., 2016. Pamidronate functionalized nanoconjugates for targeted therapy of focal skeletal malignant osteolysis. Proc Natl Acad Sci USA. 113, E4601-9. doi: 10.1073/pnas.1603316113. Epub 2016 Jul 25. PMID:27457945

Zhang, H., Du, M., Yang, Q., and Zhu, M.J., 2016. Butyrate suppresses murine mast cell proliferation and cytokine production through inhibiting histone deacetylase. Journal of Nutritional Biochemistry, 27: 299-306.

Zhou, H.R., Pestka, J.J., 2015. Deoxynivalenol (vomitoxin)-induced cholecystokinin and glucagon-like peptide-1 release in the stc-1 enteroendocrine cell model is mediated by calcium-sensing receptor and transient receptor potential ankyrin-1 channel. Toxicol Sci 145, 407-417.

Zhu, H., Du, M., Fox, L., and Zhu, M.J., 2016. Bactericidal effects of Cinnamomum cassia oil against bovine mastitis bacterial pathogens. Food Control, 66: 291-299.