Progress 04/02/09 to 04/01/14
Outputs
Progress Report Objectives (from AD-416): The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416): Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. The following summarizes the progress of the project plan completed in September 2014. BHNRC scientists continue to study phytochemicals using cell model systems to provide critical information to understand how a phytochemical works to prevent chronic diseases such as cancer. Collaborating with the Southern Regional Research Center (SRRC), the effects of soy-derived glyceollins on androgen-responsive and non- responsive prostate cancer cells were studied. Glyceollins inhibited LNCaP cell growth similar to that of the soy isoflavone genistein, inhibited G1/S cell cycle progression, and down-regulated mRNA levels of androgen responsive genes. Effects of glyceollins on androgen responsive genes appeared to be mediated through modulation of an estrogen-mediated, but not androgen-mediated pathway. Information on glyceollin serves as the basis for a proposed in-vivo study in a new project plan 1235-51530-056-00D. Screening diet-derived compounds for anti-cancer activity led to a focus on 5 diet-derived stilbene family phytoalexins structurally similar to resveratrol, a candidate cancer-protective phytoalexin. These compounds exert differential effects on LNCaP cell-cycle. Trans-resveratrol trimethylether appeared as the most potent compound among the stilbenes tested. Treatment with trans-resveratrol trimethylether resulted in G2/M blockage, inducing apoptosis, induction of the cyclin-dependent kinase inhibitor 1A and B mRNA levels. These compounds also inhibit both androgen-mediated as well as estrogen-mediated pathways. These results provide mechanistic information on potential anti-prostate cancer activity of resveratrol and its methylether analogs. Inflammation has an important role in prostate tumorigenesis. The recruitment of inflammatory monocytes to the tumor site is precipitated by increased levels of the chemo-attractant C-C chemokine ligand 2 (CCL2). Dihydrotestosterone (DHT) induced an increase in CCL2 mRNA levels, secretion of CCL2 protein in LNCaP cells and promoted human monocyte THP- 1 cell migration. This effect of DHT was mediated through an androgen receptor (AR)-dependent pathway. Broccoli-derived cancer protective compounds, indole-3-carbinol (I3C) and 3�, 3�-diindolylmethane (DIM), inhibited promotional effects of DHT on CCL2 expression and monocyte migration in-vitro. Results suggest androgen-mediated inflammatory process can be modulated by broccoli-derived compounds. This information serves as basis for proposed in-vivo studies on cell- cell interaction in a new project plan 1235-51530-056-00D. The effects of selected phytochemicals from cruciferous vegetable such as broccoli (DIM) and watercress (PEITC) were examined in vitro using human estrogen receptor (ER) positive breast cancer cell MCF-7. Treatment of MCF-7 cell with DIM and PEITC led to inhibition of cell growth. When adding both compounds together additive effect on cell growth was observed. The mechanisms underlying the growth inhibitory effect may be due to suppression of ER mRNA expression. Hence, the phytochemical may act on ER to provide protective effects for breast cancer. A grape-derived phytochemical resveratrol, and soy-derived phytochemicals genistein and glyceollins induce Liver X receptor (LXR)-responsive ABCA1 and ABCG1, and the efflux pump of intracellular cholesterol, mRNA. These results support the possibility of increased removal of cholesterol from cells by phytochemicals, but the mechanisms appear indirect through inhibition of AR-mediated pathway. Since cholesterol is associated with promotion of prostate cancer, this may be a potential mechanism for the protective effect of phytochemicals. The effects of phytochemicals were also examined using the human macrophage THP-1 cell culture model. Minimal effects by I3C, DIM, PEITC, resveratrol, genistein, glyceollins on ABCA1 and ABCG1 expression were found. Additionally, genistein did not affect LXR-responsive genes in pig-derived macrophages. By contrast, glyceollins were found to induce ABCG1 but not ABCA1 in the human hepatic cell line HepG2. These results suggest that regulation of the LXR- dependent pathway may be complex and dependent on compounds and target tissues. As the incidence of obesity reaches epidemic proportions, interest in the impact of dietary components on body-weight and fat deposition grows. Diet composition and associated microbial communities were evaluated. The anti-inflammatory protein tristetraprolin (TTP) was analyzed after feeding probiotics to pigs and significant changes related to regulation of adipocyte numbers, size, or fat accumulation were not detected. The probiotic bacteria L. paracasei possesses cholesterol lowering and immune modulating properties when added to the diet. Alveolar macrophages (AM) isolated from pigs fed an HF diet had significantly higher concentrations of cholesterol esters compared to AM from pigs fed a control (C) basal diet, but not in pigs fed a high fat (HF) diet and given L. paracasei. Ex vivo stimulation of AM with LPS significantly opposed LXR agonist- mediated transcription of cholesterol metabolism genes and abrogated the effect in AM from pigs given L. paracasei. Protein analysis revealed that AM from HF/L.paracasei-fed pigs had significantly lower LPS-induced protein expression of IL-1� than AM from HF-fed pigs. Moreover, AM from pigs fed the C diet and given L. paracasei had significantly higher mRNA levels of IL-8, 6, and 10 in response to LPS. Hence, L. paracasei modulates AM cholesterol metabolism and the response to LPS. Dietary fructose relates to obesity and has thus received considerable public attention. Juvenile pigs modeling metabolic changes similar to adolescent humans consuming fructose were examined. High fat diets increased body weight gain, total body fat deposition, elevated expression of inflammatory genes in liver, increased leptin gene expression in visceral and peripheral adipocytes, and increased expression of macrophage associated markers in visceral adipose tissue stromal cells. Pigs that consumed 20% of calories from fructose in a basal diet or combined with a high fat diet for 12 weeks showed a non- significant increase in weight gain and total body fat with a significant increase in systemic endotoxin levels but no change in inflammatory parameters measured in liver, visceral fat stromal cells, or hippocampus. There was an increase in mean visceral adipocyte area when high fructose was included in the diet. These results suggest that a moderately high amount of fructose contribute to obesity-related changes in adipocyte morphology, endotoxin level, and body weight. A prolonged consumption of fructose may be required to observe obesity-induced inflammatory changes. To define the effect of dietary fat on body weight, host adiposity, and intestinal microbiota composition, the effect of adding different fat sources (palm oil vs anhydrous milk fat-AMF) was studied. Total body weight, fat deposition, and lipid profile parameters were not affected by consumption of the type of fat, however total lean content significantly increased in high fat (HF)-AMF pigs compared to basal fat (BF) AMF. Expression of genes associated with fatty acid metabolism in liver reduced in HF diet groups independent of type of fat. Expression of SREBP1, a transcription factor responsible for regulating genes for de novo lipogenesis was up-regulated in liver of pigs fed HF palm oil only. Transcriptomic analysis of liver revealed that pigs fed the HF-palm oil diet presented 16 times more differentially expressed genes than pigs fed the HF-AMF diet. Compared to their BF level control group, amino acid metabolism, and cholesterol uptake genes were differentially affected in pigs fed HF-palm oil. Metabolomic analysis of livers indicated that pigs consuming the HF-AMF diet exhibited significantly higher levels of multiple essential fatty acids including eicosapentaenoate (EPA=20:5n3). Liver metabolite profile also indicated that pigs consuming the HF-palm oil diet had reduced amino acid related metabolites confirming functional predictions from liver transcriptomic data. Visceral adipocyte morphometric analysis indicated a reduction in visceral adipocyte numbers and increased visceral adipocyte mean area in pigs consuming HF diets and an increase in mean visceral adipocyte area in pigs that consumed HF-palm oil. Microbiota analysis indicated discrete changes in certain bacterial families. The lack of a detectable effect of broccoli-derived compounds on the LXR- dependent pathway, the availability of a well-characterized quantity of cranberry and grape seed extract sufficient to feed a large group of pigs, resulted in a redirection of the work to study cranberry/grape seed phytochemicals. Basic knowledge was accumulated on the effects of consuming proanthocyanidins in a pig model on epithelial cell function, changes in intestinal microbiome and function, and inhibition of bacterial pathogens on epithelial cell surfaces. A manuscript describing the results of the study is in preparation. The studies on phytochemicals and cancer models in vitro are also part of agreement 1235-51530-056-05S, 1235-51530-056-04R, and 1235-51530-056-06S. The work using pig model is also part of agreements 1235-51530-056-03S, 1235-51530-053-11T, 1235-51530-056-08H and 1235-51530-056-07I. In collaboration with scientists from the University of Maryland College Park, the utility of nanotechnology in stabilizing/protecting degradation of phytochemical, to enhance bioavailability and efficacies for phytochemicals was examined. A primary focus of these studies was folic acid as a specific targeting molecule, and the use of corn protein zein as packaging material for nano-particles. Future study will examine the utility of folic acid and zein in an in vivo delivery system. Accomplishments 01 Source of dietary fat affects liver transcriptome and metabolome in pigs. RNA sequencing revealed that pigs fed an equi-caloric high fat- palm oil diet presented 16 times more differentially expressed genes (240) than pigs fed the high fat anhydrous milkfat diet (15) when compared to their basal fat level control group. Data mining analysis indicated that aminoacid metabolism and cholesterol uptake were functions compromised in pigs fed the highfat-palm oil. Interestingly, expression of SREBP1, a transcription factor responsible for regulating genes required for de novo lipogenesis was up-regulated in the liver of pigs fed high fat palm-oil with no change in pigs fed the diet. Metabolomic analysis of liver samples supported the liver transcriptomic data as only pigs consuming the highfat-palm oil diet had a reduction in amino acid-related metabolites. 02 Increased dietary fructose induced changes in adipocyte morphology, endotoxin levels, and weight gain in pigs. A continued 12-week fructose consumption (20% of calories) diet in juevenile pigs induced changes in body weight, systemic endotoxin levels, and visceral adipocyte mean area with no changes in systemic inflammatory markers. This study suggested that a prolonged exposure to high levels of fructose during growth induce changes that may predispose to an obesogenic phenotype. 03 Corn protein zein can be used as packaging material for nano-particle fabrication. This provides added value for corn utilization.
Impacts
(N/A)
Publications
- Teng, Z., Luo, Y., Wang, T.T., Zanh, B., Wang, Q. 2013. Development and application of nanoparticles synthesized with folic acid-conjugated soy protein. Journal of Agricultural and Food Chemistry. 61(10):2556-64.
- Trasino, S.E., Dawson, H.D., Urban Jr, J.F., Wang, T.T., Solano Aguilar, G. 2013. Feeding probiotic Lactobacillus paracasei to Ossabaw pigs on a high fat diet prevents cholesteryl-ester accumulation and LPS modulation of the Liver X receptor and inflammatory axis in alveolar macrophages. Journal of Nutritional Biochemistry. 24(11):1931-1939.
- Fayer, R., Elsasser, T.H., Gould, R., Solano Aguilar, G., Santin, M., Urban Jr, J.F. 2014. Blastocystis tropism in the pig intestine. Parasitology Research. 113:1465-1472.
- Huang, H., Xie, Z., Boue, S.M., Bhatnagar, D., Yokoyama, W.H., Yu, L.L., Wang, T.T. 2013. Cholesterol-lowering activity of soy-derived glyceollins in the golden Syrian hamster model. Journal of Agricultural and Food Chemistry. 61(24):35772-5782.
- Luo, Y., Teng, Z., Wang, T.T., Wang, Q. 2013. The cellular uptake and transport of zein nanoparticles: Effect of sodium caseinate. Journal of Agricultural and Food Chemistry. 61(31):7621-9.
- Panickar, K.S., Jang, S. 2013. Dietary and plant polyphenols exert neuroprotective effects and improve cognitive function in cerebral ischemia. Recent Patents on Food, Nutrition and Agriculture. 5(2):128-43.
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Progress 10/01/12 to 09/30/13
Outputs
Progress Report Objectives (from AD-416): The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416): Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. In vitro and in vivo models have been used to identify biologically active compounds from the diet. Using a prostate cancer cell model in- vitro, we observed that soy glyceollins inhibited LNCaP prostate cancer cell growth similar to that of the soy isoflavone genistein. The growth inhibitory effects of the glyceollins appeared to be due to an inhibition of cell cycle progression. In addition, glyceollin treatments led to down- regulated mRNA levels for androgen responsive genes. We began to characterize Liver X receptors (LXR)-dependent pathways in-vitro by cancer protective phytochemicals. Expression of LXR-responsive genes is being used as markers for potential modulation of the pathway. We also initiated a study to examine whether cancer protective effects of broccoli-derived phytochemicals can be synergistic or additive. LXR regulate cholesterol metabolism and inflammatory responses. High fat (HF) diets and microbial infection antagonize the LXR pathway leading to accumulation of cholesteryl-esters (CE) and increased expression of pro- inflammatory mediators in macrophages. Alveolar macrophages (AM) isolated from Ossabaw pigs fed an HF diet had significantly higher concentrations of CE compared to AM from pigs fed a control (C) basal diet, but not in pigs fed a HF diet and given the probiotic L. paracasei (HFPB). Ex vivo stimulation of AM with LPS significantly opposed LXR agonist-mediated transcription of cholesterol metabolism genes in pigs. This transcriptional effect was abrogated in AM from pigs given L. paracasei. Protein analysis of cell culture supernatants revealed that AM from HFPB- fed pigs had significantly lower LPS-induced protein expression of IL-1� than AM from HF-fed pigs. These data demonstrated a role for L. paracasei in modulating AM cholesterol metabolism and the response to LPS. The anti-inflammatory protein tristetraprolin (TTP) was analyzed and we did not observe significant changes related to regulation of adipocyte numbers, size or fat accumulation by probiotics. Hence, a large-scale non- targeted metabolomic approach was used to reveal metabolite patterns from obesity-related metabolic disturbances in a pig model. Feeding a high energy/high fat diet influenced metabolic homeostasis of Ossabaw pigs throughout all tissues and biofluids examined. Metabolite profiling involving multiple sample types was shown to be a feasible method to provide information on metabolic pathways affected by the dietary treatment. Proanthocyanidins, which are amongst the most bioactive polyphenols in the diet, are catabolized by intestinal microbiota. The effect of cranberry extract containing ~7% proanthocyanidins (ECPA), on weight gain and composition of intestinal microbiota was evaluated in pig model. Young pigs fed ECPA added to a basal or high fat diet for 12 weeks were compared to pigs fed similar diets without ECPA. DNA from fecal samples, collected at day 0 and weeks 2, 4, 6, 10, and contents from proximal and distal colon collected at necropsy were processed. Molecular and metabolic analyses are currently underway. Accomplishments 01 Identify plant bioactive components and their mechanisms of action. The anti-cancer effects of a broccoli-derived compound indole-3-carbinol and diindolylmethane was examined using an in-vitro model of prostate cancer. These compounds were found to indirectly affect monocyte/ macrophage migration by regulating the production of chemo-attractants of monocyte/macrophage by prostate cancer cells. Inflammation is an important promoter of tumor cell invasion and metastasis. This information provides mechanistic support for the potential beneficial effects of cruciferous vegetables on prostate cancer development. 02 Conducted a large scale metabolomic analysis of pigs on a high energy/ high fat diet. Typical analyses of body fluids from human dietary interventions do not provide adequate information about diet-induced metabolic changes taking place in tissues. We used a large-scale, non- targeted metabolomic approach to reveal metabolite patterns from obesity-related metabolic disturbances produced in a specialized pig model. Feeding a high energy/high fat diet influenced metabolic homeostasis of Ossabaw pigs throughout all tissues and biofluids including plasma, urine, bile, liver, pancreas, brain cortex, intestinal jejunum, and proximal colon. Metabolite profiling involving multiple sample types was shown to be a feasible methodology to provide information on metabolic pathways affected by the dietary treatment. This study provides a more comprehensive evaluation of the effect of diet on metabolism of the whole animal that can be used to better understand the role of diet in health. 03 Daily feeding of probiotics can improve the activity of immune cells in the lungs. High fat diets and microbial infection can antagonize important pathways in inflammatory cells in the lung called macrophages that normally accumulate cholesterol from the blood and release pro- inflammatory molecules to control infection. Feeding the probiotic bacteria Lactobacillus paracasei can lower blood cholesterol and modulate protective immune function. We used this information to demonstrate that daily feeding of L. paracasei to a unique variety of Ossabaw pigs that become obese when fed a high fat diet could modulate cholesterol metabolism and improve macrophages immune capacity. This is one of the few demonstrations of the beneficial effects of adding probiotics to the diet to improve metabolism and immune function in the lung. 04 Infection with parasitic worms can improve metabolic function in obese mice. Obesity is associated with a chronic low-grade inflammation and increased levels of pro-inflammatory molecules that can disrupt metabolic homeostasis. Parasitic worm infection has been shown to reduce the intensity of certain inflammatory diseases. We examined the effects of worm infection in a mouse model of obesity and associated metabolic dysfunction to explore the mechanism of action and determine if features of the interaction could be used to control obesity related inflammation. The infection reduced body weight and fat tissue mass in mice fed a high fat diet and improved glucose metabolism along with healthy changes in the profile of metabolic hormones. The results show that parasitic worms have both preventive and therapeutic effects against the development of obesity and associated features of metabolic dysfunction in mice. Although not directly useful as a treatment strategy in humans, it does provide a model to look for the molecules produced by worms that could be applied in humans to reduce inflammatory diseases.
Impacts
(N/A)
Publications
- Hanhineva, K., Barri, T., Kolemainen, M., Pekkinen, J., Pihlajamaki, J., Vesterbacka, A., Solano Aguilar, G., Mykk�nen, H., Dragsted, L., Urban Jr, J.F., Poutanen, K. 2013. Comparative non-targeted metabolite profiling of metabolic changes in tissues and bio-fluids in high-fat diet fed Ossabaw pig. Journal of Proteome Research. DOI: 10.1021/pr400257d.
- Yang, Z., Grinchuk, V., Smith, A.D., Qin, B., Bohl, J.A., Sun, R., Notari, L., Zhang, Z., Sesaki, H., Urban Jr, J.F., Shea-Donahue, T., Zhao, A. 2013. Parasitic nematode-induced modulation of body weight and associated metabolic dysfunction in mouse models of obesity. Infection and Immunity. 81(6):1905-1914.
- Huang, H., Fletcher, A., Niu, Y., Wang, T.T., Yu, L. 2012. Characterization of lipopolysaccharide-stimulated cytokine expression in macrophages and monocytes. Inflammation Research. 61(12):1329-1338.
- Kim, E.K., Kim, Y.S., Milner, J.A., Wang, T.T. 2013. Indole-3-carbinol and 3�, 3�-diindolylmethane modulate androgen effect up-regulation on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells. Cancer Prevention Research. 6(6):519-529.
- Wang, T.T., Edwards, A.J., Clevidence, B.A. 2013. Strong and weak plasma response to dietary carotenoids identified by cluster analysis and linked to beta-carotene 15,15'-monooxygenase 1 single nucleotide polymorphisms. Journal of Nutritional Biochemistry. 24(8):1538-1546.
- Lou, Y., Wang, T.T., Teng, Z., Chen, P., Sun, J., Wang, Q. 2013. Encapsulation of indole-3-carbinol and 3'3'-diindolylmethane in zein/ carboxymethyl chitosan nanoparticles with controlled release property and improved stability. Food Chemistry. 139(1-4):224-30.
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416): Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. Continued use of in vitro and in vivo models identified biologically active compounds from the diet to test their role in prevention of chronic diseases such as obesity and cancer and to elucidate mechanisms of action. Our mechanistic studies focused on androgen receptor and liver X receptor (LXR)-mediated pathways. Androgen exposure is a risk factor for prostate cancer. The LXR receptor is responsible for cholesterol removal and is important for prevention of cardiovascular disease and prostate cancer. Glyceollins are soy-derived compounds that have been proposed to be candidate cancer preventive compounds. The effect of the glyceollins on prostate cancer is unknown. We found that the glyceollins inhibited the growth of a prostate cancer cell line. This effect of glyceollins appears to be mediated through modulation of an estrogen- but not androgen-mediated pathway and is different from other soy-derived phytochemicals. Similarly, compounds, such as indoles and thiocyanates, found in cruciferous vegetables, such as broccoli, were identified as compounds that exert inhibitory effect on breast cancer and prostate cancer cell growth in vitro. In prostate cancer cells, these compounds appeared to act independently of the LXR pathway by inhibiting an androgen receptor mediated pathway. Given the lack of effect of broccoli- derived compounds on LXR pathways, as a contingency, we examined cranberry and grape seed-based phytochemicals in a pig model. Phytochemicals from these plants have been reported to affect LXR- mediated pathways in vitro and reduce adiposity in rodent high-energy diet feeding study. These effects are currently being evaluated in vivo in a pig high-energy diet feeding study. A pig model was used to study the relationship between diet and juvenile obesity. Excess high-fructose (HFT) combined with a high-fat (HF) diet were found to increase central obesity and body weight gain in pig. These changes in fat mass were accompanied by increased total bacterial counts when compared to pigs fed basal diets. High-fructose diets induced a decrease in Bifidobacterium species and increased total coliforms. Bifidobacterium species are potentially beneficial bacteria while coliforms negatively affect intestinal health. This information provides evidence that excess calories from HFT diets exacerbate certain features of the obesogenic phenotype through increased lipidogenic activity, peripheral fat deposition, and modified intestinal bacterial populations. Accomplishments 01 The anti-cancer effects of a broccoli-derived compound phenethyl isothiocyanate was examined using a mouse model of prostate cancer. The compound was found to indirectly affect tumor growth through inhibition cell attachment factors and new blood vessel formation. Using high- through-put DNA sequence analysis technology, it was further determined that several markers of inflammation were reduced. Inflammation is an important promoter of tumor cell invasion and metastasis. This information provides mechanistic support for the potential beneficial effects of cruciferous vegetable on prostate cancer development. 02 Obesity characterized by weight gain, fat deposition, and induction of inflammatory responses has been reproduced in young pigs as a large anim model for childhood obesity. A study of the effects of high-fat (HF) and/or high-fructose (HFT) diets on molecular changes in the brain of young pigs revealed that a HF, but not a HFT diet, increased pro- inflammatory cytokine expression in the area involved in memory. This information suggested that dietary contributions of excess calories to brain dysfunction should be examined separately. It also provided an additional target organ for evaluating the negative effects of excessive caloric intake.
Impacts
(N/A)
Publications
- Palavalli, M.H., Natarajan, S.S., Wang, T.T., Krishnan, H.B. 2012. Inhibition of soybean seeds in warm water results in the release of copious amounts of Bowman-Birk protease inhibitor, a putative anticarcinogenic agent. Journal of Agricultural and Food Chemistry. 60:3135-3143.
- Wang, T.T., Boue, S.M., Krishnan, H.B. 2011. The protective effect of soybean phytochemicals on androgen responsive human prostate cancer cells LNCaP is likely mediated through modulation of hormone/cytokine-dependent pathways. Functional Foods in Health and Disease. 1(11):457-471.
- Shea-Donohue, T., Notari, L., Urban Jr, J.F., Zhao, A. 2010. Immune regulation of epithelial cell function: Implications for GI pathologies. International Dairy Journal. 20(4):248-252.
- Li, R.W., Li, C., Wang, T.T. 2012. Transcriptomic alterations in human prostate cancer cell LNCaP tumor xenograft modulated by dietary phenethyl isothiocyanate. Molecular Carcinogenesis. DOI: 10.1002/mc.21873.
- Wu, M., Wu, R., Wang, T.T., Cheng, W. 2011. A role for p53 in selenium- induced senescence. Journal of Agricultural and Food Chemistry. 591:1182- 1187.
- Butler, J.E., Sun, X., Wertz, N., Lager, K.M., Chaloner, L., Urban Jr., J., Francis, D.L., Nara, P.L., Tobin, G.J. 2011. Antibody repertoire development in fetal and neonatal piglets XXI. Usage of most VH genes remains constant during fetal and postnatal development. Molecular Immunology. 49(3):483-494.
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416) Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. We hypothesized that modulation of androgen and/or cholesterol-related pathways by cruciferous vegetable-derived compounds indole-3-carbinol (I3C) and diindolylmethane (DIM) may be potential mechanisms by which these bioactive food components exert their protective effects against prostate cancer. We focused on the effects of I3C and DIM on Liver X receptors (LXRs)-mediated pathways. LXR-alpha and beta are transcriptional factors that use cholesterol metabolites oxysterol as ligands and serve as critical regulators of cholesterol metabolism. It was found that DIM, but not I3C, induces the transcripts of two LXR- responsive ATP-binding cassette transporters involved in cholesterol efflux, ABCG1 and ABCA1 mRNA, in the androgen responsive human prostate cancer cell LNCaP. Exposure of LNCaP cells to 5 microM DIM for 48 hours lead to a 5-fold increase in ABCG1 and ABCA1 mRNA levels. Knock-down of LXR expression using siRNA against both alpha and beta isoforms abrogated the ability of DIM to induce ABCG1 but not ABCA1 mRNA, supporting the effects of DIM on ABCG1 is mediated through LXR-associated pathway. However, we did not observe induction of ABCG1 or ABCA1 mRNA in androgen non-responsive human prostate cancer cells PC3 treated with DIM. Furthermore, in the LNCaP cells, knock-down of the androgen receptor (AR) expression with AR siRNA blocked DIM induction of ABCA1 and ABCG1 mRNAs. In summary, our results suggest that DIM may play a role in modulating cholesterol metabolism, in part via androgen-mediated pathway in prostate cancer cells. However, since DIM modulation of ABCG1 was LXR-dependent, it is likely that multiple pathways are involved with DIM modulation of cholesterol metabolism. Furthermore, these effects are unique to DIM, but not I3C, suggesting these two dietary components may act through different mechanisms for their cancer preventive effects. To further the understanding of how phytochemicals may prevent cancer, procedures were expanded to detect how these nutrients effect the regulatory proteins that control growth in both normal and tumor cells. The phytochemicals tested in our model cellular systems, such as resveratrol, have been shown to be more effective in modulating the regulatory proteins in cancer cell lines rather than in normal cell lines. By observing changes in proteins such as p21, p27, hypophosphorylated Rb, and cyclins B,D, and E, these procedures can help characterize the potential for anti-cancer of phytochemicals. Accomplishments 01 With the advent of high-throughput sequencing methods, there is an increased interest in the fields of metagenomics and microbiomics. Often researcher is interested in knowing which features, such as operational taxonomic units and clusters of orthologous genes, in a microbiome or metagenome dataset are differentially abundant. It can be difficult, however, for a researcher to know which statistical method is most appropriate for a particular dataset. This study compared the suitabili of several statistical methods for detecting differentially abundant features in high-throughput microbiome and metagenome datasets. Amongst the statistical methods tested, an arcsine square root data transformati followed by the t Test (one-factor case), Gaussian generalized linear model (one- and two-factor cases), or ANOVA Type II sum of squares (2- factor case) are recommended for high-throughput microbiome and metageno data quantified as relative abundances. 02 There has been growing interest in examining the immune-modulating effec of some strains of lactic acid producing probiotic bacteria on the gut micro-biota. Data from human and animal studies have established that so commensal derived probiotic bacteria can decrease expression of pro- inflammatory mediators and lower serum cholesterol levels by decreasing entrohepatic recirculation of bile acids and cholesterol. There is littl characterization of the systemic effects of probiotics on cholesterol metabolism in extra-hepatic tissue. Recent evidence suggests that probiotics can modulate cholesterol absorption through interactions with LXRa and NPC1L implicating effects on the LXR axis in cell types outside of the gut. We, therefore, examined whether alveolar macrophages (AM) isolated from lungs of juvenile Ossabaw pigs fed either a high fat (HF) HF diet supplemented with the probiotic (PB) Lactobacillus paracasei changed cholesterol metabolism and expression of LXR and inflammatory related genes and proteins. AM isolated from pigs fed the HF diet had significantly higher concentrations of cholesterol esters (CE) compared with AM from pigs fed a control basal diet, but this effect was not observed in AM from pigs fed a HF diet given a daily treatment with PB. vivo stimulation of AM with LPS significantly opposed LXR agonist-mediat transcription of cholesterol metabolism genes. This effect was abrogated for ABCA1, CH25H, and PPARy expression in AM from pigs fed PB. We observed that AM from pigs fed a HF diet had a significantly higher IL-1 mRNA response to LPS. This was not affected by PB. Moreover, AM from pigs fed a control diet supplemented with PB had significantly higher mR levels of IL-6 in response to LPS. Taken together, these data demonstrated a role for probiotic L. paracasei in modulating AM cholesterol metabolism and response to inflammation. 03 Food intake and energy expenditure are regulated by the central nervous system. Specifically, the hypothalamus plays an important role in the regulation of satiety and appetite through leptin and insulin signaling. Leptin, a cytokine secreted by adipocytes, is known to reduce food intak and increase energetic expenditure. Hyperleptinemia, an excess of leptin production, and leptin resistance, a decreased response to exogenous leptin, have been observed in obesity. Although many obesity studies revealed important mechanisms of leptin resistance in hypothalamus, most of the studies have been done in small animal models such as mice and ra Compared to the rodent brain, the pig brain more closely resembles the human brain in terms of both anatomy and biochemistry, which associates the pig with a higher translational value. We recently established an ex vivo functional assay using sections of pig brain to examine leptin response. Brain slices from pig hypothalamus were cultured with or witho human recombinant leptin and measured for phosphorylation of STAT3, whic is considered as an indicator of leptin signaling in vivo. A dose- dependent increase of p-STAT3 was found with increasing levels of leptin added to hypothalamus. These preliminary results demonstrated that short term ex-vivo culture of hypothalamus sections constitutes an ex-vivo functional assay to examine leptin/insulin signaling in brain from pigs fed different obesogenic diets. Currently, juvenile pigs were fed three different obesogenic diets containing excess calories from a fructose, f or a combination of high fat and fructose, and brain sections from this study will be used to investigate the effect of diet composition on lept signaling. 04 Five Gynostemma pentaphyllum (GP) samples were investigated and compared for their chemical compositions and their antioxidant, antiproliferative and anti-inflammatory effects. Extracts (50% acetone, 75% ethanol, and 100% ethanol) of the five GP samples (GP1-5) differed in their total phenolic, saponin, and flavonoid contents and in their rutin and quercet concentrations. The highest level of total flavonoids was 63.5 mg of rut equiv/g in GP4, and the greatest total phenolic content was 44.3 mg of gallic acid equiv/g in GP1 with 50% acetone as the extraction solvent. G had the highest total saponin content of 132.6 mg/g with 100% ethanol as the extraction solvent. These extracts also differed in their scavenging capacity against DPPH and hydroxyl radicals, although they all showed significant radical scavenging capacity. The 100% ethanol extracts also showed dose dependent inhibition of IL-6 and Ptgs2 mRNA expression and weak inhibition on TNF-R mRNA expression. In addition, GP1 had the highe antiproliferative activity at 3.2 mg equiv/mL concentration in HT-29 hum colon cancer cells. The results will be used to study the use of product derived from G. pentaphyllum for improving human health. 05 Engeletin and astilbin, flavonoid compounds, were isolated from the leav of Engelhardia roxburghiana for the first time. The chemical structures engeletin and astilbin were confirmed by 1H and 13C nuclear magneticresonance (NMR) and mass spectrometry (MS) spectra, and their an inflammatory activities were studied in lipopolysaccharide (LPS)- stimulated mouse J774A.1 macrophage cells. LPS induced the inflammatory state in macrophage cells and increased mRNA expressions of pro- inflammatory cytokines. Engeletin and astilbin exhibited remarkable inhibitory effects on interleukin (IL)-1� and IL-6 mRNA expression. Significant inhibition of LPS-mediated mRNA expressions were also seen i LPS binding toll-like receptor (TLR)-4, pro-inflammatory cytokine tumor necrosis factor (TNF)-R, IL-10, chemoattractant monocyte chemotactic protein (MCP)-1, and cyclooxygenase (COX)-2 genes. The reduced expressio of these cytokines may modulate the immune response and reduce inflammatory activation, indicating that engeletin and astilbin may serv as potential anti-inflammatory agents.
Impacts
(N/A)
Publications
- Huang, H., Cheng, Z., Shi, H., Xin, W., Wang, T.T., Yu, L. 2011. Isolation and characterization of two flavonoids, engeletin and astilbin, from the leaves of Engelhardia roxburghiana, and their potential anti-inflammatory properties. Journal of Agricultural & Food Chemistry. 59(9):4562-9.
- Wang, T.T., Schoene, N.W., Milner, J.A., Kim, Y.S. 2011. Broccoli-derived phytochemicals indole-3-carbinol and 3,3�-diindolylmethane exert concentration-dependent pleiotropic effects on prostate cancer cells: Comparison with other cancer preventive phytochemicals. Molecular Carcinogenesis. DOI: 10.1002/mc.20774.
- Xie, Z., Liu, W., Huang, H., Slavin, M., Zhao, Y., Whent, M., Blackford, J. , Lutterodt, H., Zhou, H., Chen, P., Wang, T.T., Liangli, Y. 2010. Chemical composition of five commercial gynostemma pentaphyllum samples and their radical scavenging, antiproliferative, and anti-inflammatory properties. Journal of Agricultural and Food Chemistry. 58(21):11243-11249.
- Qi, Y., Schoene, N.W., Lartey, F., Cheng, W. 2011. Selenium compounds activate ATM-dependent DNA damage responses via the mismatch repair protein hMLH1 in colorectal cancer cells. Journal of Biological Chemistry. 285(43):33010-33017.
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416) Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. Feeding a high fat/high fructose/high cholesterol (HF/HFr/HC) diet to juvenile Ossabaw pigs induced obesity and insulin resistance, visceral adipose expansion, hypertriglyceridemia, and increased LDL:HDL and serum cholesterol levels indicative of metabolic syndrome. Inflammation related to obesity contributes to poor health. Certain strains of probiotic bacteria, including Bifidobacterium lactis Bb12 (Bb12), reduce inflammation. To test the hypothesis that Bb12 could reduce inflammation, three groups of Ossabaw pigs were maintained on 1) a HF/HFr/HC diet and fed daily 2 x 1010 cfu of Bb12, 2) a HF/HFr/HC diet and fed a probiotic-free placebo, or 3) maintained on a control basal diet. The HF/HFr/HC diet induced a significant increase in intestinal coliform bacteria and a decrease in Bifidobacterium species in feces compared to pigs on the basal diet, and this effect was reversed in pigs fed the probiotic. Analysis of the intestinal microbiota by molecular pyrosequencing showed diet-dependent shifts in bacterial populations. Analysis of the local host response showed a 7-fold increase in leptin gene expression in adipocytes isolated from pigs fed the HF/HFr/HC diet and given the placebo that was absent in pigs fed the probiotic. Differences in leptin receptor gene expression in the brain were also observed. In a separate study, diet-derived compounds were screened for anti- cancer activity using cell culture-based models. Several phytoalexins, plant phytochemicals activated by stress, inhibited the androgen- responsive human prostate cancer cell line LNCaP. Five diet-derived phytoalexins structurally similar to resveratrol, a cancer protective phytoalexin found in grape skin, exerted differential effects on LNCaP cell growth, cell cycle, and programmed cell death (apoptosis). A derivative compound, trans-resveratrol trimethylether, was the most potent among those tested because it blocked LNCaP cell cycle (G2/M) and induced apoptosis. The effects of these compounds on cell cycle correlated with induction of the cyclin-dependent kinase inhibitor 1A and B mRNA levels, inhibited both androgen- and estrogen-mediated signaling pathways, and provided mechanistic information on how resveratrol and its analogs contribute to anti-prostate cancer cell activity. In another cell culture study, the effects of phytochemcials on the liver-x-receptor (LXR) responsive genes ABCA1 and ABCG1 that regulate cellular cholesterol efflux pumps were examined. Several phytochemicals, including resveratrol, soy-derived genistein and glyceollins, and broccoli-derived 3�, 3� diindolylmethane, induced ABCA1 and ABCG1 mRNA, supporting the hypothesis that phytochemicals increase the removal of cholesterol from cells. Because cholesterol is a promoter of prostate cancer and obesity-related inflammation, this observation helps explain the health protective effect of phytochemicals. Accomplishments 01 The current research addressed the lack of molecular and mechanistic information on efficacy of the cancer preventive phytochemicals such as resveratrol and its analogues. Five diet-derived phytoalexins structural similar to resveratrol, a cancer protective phytoalexin found in grape skin, exerted differential effects on LNCaP cell growth, cell cycle, and programmed cell death (apoptosis). Several phytoalexins, plant phytochemicals activated by stress, inhibited the androgen-responsive human prostate cancer cell line LNCaP. A derivative compound, trans- resveratrol trimethylether, was the most potent among those tested becau it blocked LNCaP cell cycle (G2/M) and induced apoptosis. The effects o these compounds on cell cycle correlated with induction of the cyclin- dependent kinase inhibitor 1A and B mRNA levels, inhibited both androgen and estrogen-mediated signaling pathways, and provided mechanistic information on how resveratrol and its analogs contribute to anti-prosta cancer cell activity. The results support the hypothesis that phytochemicals interact with multiple cellular pathways important in cancer development and regulate cancer cell growth, cell cycle, and apoptosis, providing information that diet-derived phytochemicals can serve as potent inhibitors of carcinogenesis. 02 The Ossabaw pig study represents a proof of principle that juvenile Ossabaw pigs fed a HF/HFr/HC diet can rapidly develop features of the metabolic syndrome and that daily feeding of certain probiotics can regulate the severity of juvenile obesity. Analysis of the intestinal bacterial populations showed that certain bacterial phyla virtually disappeared from the lumen of pigs on a HF/HFr/HC diet and other phyla were expanded. Pigs fed the probiotic, however, altered the distributio of bacterial genera in both the lumen and mucosa compared to those fed t placebo. Associations between these changes in bacterial populations an inflammatory and metabolic parameters in the host can be linked to contr strategies. These studies provide a solid foundation for additional testing of diet and food-derived compounds thought to prevent or ameliorate expression of chronic disease and to determine mechanisms of action at the molecular, cellular and whole animal level.
Impacts
(N/A)
Publications
- Wu, M., Kang, M., Schoene, N.W., Cheng, W. 2010. Selenium compounds activate early barriers of tumorigenesis. Journal of Biological Chemistry. 285(16):12055-12062.
- Shih, R.S., Wong, S.H., Schoene, N.W., Zhang, J.J., Lei, K.Y. 2010. Enhanced Gadd45 expression and delayed G2/M progression are p53 dependent in zinc-supplemented human bronchial epithelial cells. Experimental Biology and Medicine. 235(8):932-940.
- Wang, T.T., Schoene, N.W., Kim, Y.S., Mizuno, C.S., Rimando, A.M. 2010. Differential effects of resveratrol and its methylether analogs on cell cycle and apoptosis in human androgen-responsive LNCaP cancer cells. Molecular Nutrition and Food Research. 54(3):335-344.
- Zhang, J., Wu, M., Schoene, N.W., Cheng, W., Wang, T.T., Alshatwi, A.A., Alsaif, M., Lei, K.Y. 2009. Effect of resveratrol and zinc on intracellular zinc status in normal human prostate epithelial cells. American Journal of Physiology - Cell Physiology. 297(3):C632-644.
- Han, C.T., Schoene, N.W., Lei, K.Y. 2009. The influence of zinc deficiency on AKT-MDm2-p53 signaling axes in normal and malignant human prostate cells. American Journal of Physiology - Cell Physiology. 297(5):C1188-1199.
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) The overall goal of the project is to elucidate the molecular and cellular mechanisms that respond to selected health promoting food components to reduce the risk of chronic diseases such as cancers and obesity. A secondary aim is to explore the utility of a porcine model to test the effect of health maintenance via diet and identify resulting biomarkers that reflect health status. Objective 1. Elucidate biological activities of health promoting phytochemicals from grape, soy, and cruciferous vegetables against development of breast and prostate cancer. Objective 2. Identify molecular targets and mechanisms of action of health promoting food components in animal or in vitro models of cancer and obesity. Objective 3. Ascertain the effects of specific probiotic strains in appropriate animal models of obesity. Objective 4. Identify plant polyphenols and probiotics that affect adipocyte numbers, size, and fat accumulation, and the regulation of proinflammatory mRNA stability by tristetraprolin. Objective 5. Tie together obesity, inflammation, and cancer mechanistically in appropriate animal or in vitro models. Approach (from AD-416) Studies will evaluate if phytoalexins structurally similar to resveratrol exerts similar anti-prostate cancer effects; if soy phytoalexin glyceollins exert anti-prostate cancer effects; if phytochemicals modulate LXR-mediated pathways in prostate epithelial cells and modulate LXR-mediated pathways in macrophage. Other studies will determine if probiotic bacterial strains differ in their protective effects against chronic diseases related to obesity; regulate adipocyte numbers, size, and fat accumulation associated with the anti-inflammatory protein tristetraprolin (TTP); if obesity alters the macrophage phenotype and function in adipose tissue, colon, breast, and prostate following increased localized inflammation; and if broccoli-derived phytochemicals modulate LXR-responsive pathways in vivo. The studies will involve in vitro cell culture approaches confirmed by rodent and pig animal models. Significant Activities that Support Special Target Populations Notice was received in April 2009 that this project was approved and thus was in place for about five months during FY2009. We continue to examine the molecular action of phytochemicals on regulation of human sex hormone receptors, estrogen receptors (ERs), and androgen receptor (AR)-mediated events, which play important roles in breast and prostate cancer development. Sex hormone receptors can influence a cellular process such as cell death or apoptosis of breast cancer cells and this process can be modulated by soy-derived phytochemicals. We demonstrated that broccoli- derived phytochemicals can exert their cancer cell-growth inhibitory effects through regulation of ER expression. This is now being extended to evaluating the effects of compounds with structural similarity to reserveratrol, a phytoalexin found in grapes and other plants, to evluate the effect on AR-mediated gene expression. Both T regulatory cells and the alternatively activated macrophage are important cells regulating responses to chronic disease. The current project plan will characterize the role of these cells at the molecular, cellular, and whole animal level in response to a high fat, high cholesterol, high refined carbohydrate diet used to induce metabolic syndrome in Ossabaw pigs and in adipose tissue and isolated stromal cells. Probiotic bacteria have beneficial effects on intestinal immune and mucosal barrier function; probiotic (Bifidobacterium lactis)-treated pigs show changes in gene expression of immune mediators indicating local activation of the innate immune system at the intestinal mucosa and induction of changes that improve intestinal permeability, a response enhanced in probiotic-treated pigs from probiotic-treated mothers. Probiotics in the diet also modulate inflammatory responses against intestinal parasites and bovine milk used as a food allergen. This research promotes the concept that probiotics in the diet differentially affect immune components associated with resolution of infectious and chronic disease. Treating juvenile Ossabaw pigs with probiotics is a logical extension of earlier studies on parasite and allergen-induced inflammation in conventional pigs.
Impacts
(N/A)
Publications
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