Exposure to environmental pollutants poses numerous risk factors for human health, including increasing incidence of cardiovascular disease and diabetes. Persistent organic pollutants, such as polychlorinated biphenyls (PCBs) have been strongly linked to the development of these chronic inflammatory diseases and the primary route of exposure is through consumption of contaminated food products. Thus, the gastrointestinal tract is susceptible to the greatest levels of these pollutants and is an important facet to study. The first two hypotheses of this dissertation tested that exposure to PCBs disrupts gut microbiota directly (in vitro) and within a whole body system. PCB exposure disrupted microbial metabolism and production of metabolites (i.e. short chain fatty acids) in vitro. These disruptions in microbial populations were consistent in our mouse model of cardiometabolic disease, where we observed reductions in microbial diversity, an increase in the putative pro-inflammatory ratio of Firmicutes to Bacteroidetes, and reductions in beneficial microbial populations in exposed mice. Furthermore, observed greater inflammation was observed both within the intestines and peripherally in PCB exposed mice as well as disruptions in circulating markers associated with glucose homeostasis. Nutritional interventions high in prebiotic dietary fiber such as inulin may be able to attenuate the toxic effects of pollutant exposure. To test the hypothesis that consumption of the prebiotic inulin can decrease PCB-induced disruption in gut microbial and metabolic homeostasis, LDLr-\- mice were fed a diet containing inulin and exposed to PCB 126. Mice fed an inulin-containing diet and exposed to PCBs exhibited improved glucose tolerance, lower hepatic inflammation and steatosis, and distinct differences in gut microbial populations. Overall, these data suggests that nutritional intervention, specifically prebiotic consumption, may reduce pollutant-induced disease risk.

Approximately 46 million acres or 88 per cent of the land area of the state of Utah has been classified by Reuss and Blanch (1951) as "range land open for grazing." Although some of this area cannot be grazed because of inaccessibility, lack of water, poison plants, and similar factors, it is estimated that some 40 million acres or 78 per cent of the total land area is available for grazing. Most of this grazing land is federally owned (73 per cent), while only 22 per cent is privately held, and 5 per cent is state owned.

The objectives of this investigation have been to characterize black gram (Phaseolus Mongo L.) and rice (Oryza sativa L.) proteins and to study changes in their nutritional value due to fermentation. black gram, the legume chosen for this work, is one of the most important legume crops throughout a large part of the tropics. The protein content of 60 mesh, dehydrated, defatted black gram meal was 28.5 percent. Sodium carbonate (0.5- 1.0 percent), tetra-sodium pyrophosphate (0.5 percent), and sodium dodecyl sulfate (SDS) (0.5-5 percent) proved to be the potential protein solubilizers as they extracted more than 76 grams of Lowry's protein per 100 grams Kjeldahl protein. On the considerations of contaminating residue in the final product and disruption of native structure of the proteins, these chemical agents were unsuitable. Sodium sulfate at the 10 percent level was judged to be the best protein solubilizer. Proteins separated on polyacrylamide gel using a phenol-acetic acid-mercaptoethanol-urea (PAMU) system were run on the flat bed gel containing SDS. The proteins were separated in 13 constituents and the molecular weights of the major ones were 140,000 and 55,000. Solubilized proteins contained 81 percent globulins, 13 percent albumins, 4 percent prolamins, and 2 percent glutelins . Sulfur containing amino acids and threonine were deficient in total proteins xv of the seeds with 27. 6 and 78.8 as their respective chemical scores . Chemical scores of the albumin, globulin, prolamin, and glutelin fractions were 64, 0, 56, and 70.7, respectively. The predicted biological values in human nutrition varied from O for globulins to 110 for glutelins, and was 14.9 for total proteins in the seeds. The constituents of the protein fractions were isoelectrically focused in the acidic pH range with the exception of two globulins for which the isoelectric points were 8.42 and 8.65. The trypsin inhibitor from black gram was isolated using affinity chromatography gel with 19. 5 fold purification. The inhibitor had 75 amino acid residues and contained one disulfide bridge. Chemical studies assigned an important role for the hydrogen bonds and demonstrated vital importance of the disulfide bridge in retaining the inhibiting activity. The inhibitor was stable and retained 35 percent of the activity when heated at 100°C for 60 minutes at pH 11. Chemical modification of amino acid residues suggested the involvement of lysine and arginine residues at the active site of the inhibitor. Lysine and arginine moieties at the active site have been proposed to be present as alanyllysine and histidylarginine. Inhib i tion of bovine pancreatic trypsin by the inhibitor was kinetically studied . The kinetic constants Km and Vm ax were 2.7 x l0- 5M and 6 x l0 - 3M/min, respectively. The dissociation constant for the enzyme-inhibitor complex (Ki) was 4 x l0- 7M, whereas that for the enzyme-inhibitor-substrate complex (K. 1 , ) was 1.89 x l0- 6M. The inhibition was a mixture of partial competitive and pure-noncompetitive systems. Rice and black gram form the integral parts of a fermented snack food of the Indian subcontinent~idli. Amino acid composition of black gram and rice were complementary. Leucine, lysine, and sulfur containing amino acids were the most limiting amino acids in rice with 65. 1, 66.3, and 67.9 as their respective scores. The estimates of biological values of rice proteins in human nutrition qualified albumins as superior and prolamins as inferior proteins. The PAMUsy stem in polyacrylamide gel electrophoresis was more efficient in resolving protein subunits than the SOS gel system. The PAMUsy stem was not sensitive to the ionic strength of the sample. Mobilities of rice and black gram proteins in SOS and PAMUsy stems were based on the related parameters. In the PAMUsy stem, the mobilities of most proteins seemed to depend on their molecular size. The PAMUsy stem on gel electrophoresis was judged superior to the SOS system. Fermentation of the black gram-rice blend was kinetically studied for changes in physicochemical characteristics and nutritional functionality. Trypsin inhibiting activity was unaffected, but chymotrypsin inhibiting activity was reduced to 3 percent after 20 hours fermentation , Significant increases were noted in the contents of sulfur containing amino acids during fermentation . These amino acids seemed to be bioavailable. In vitro digestion with pepsin and pancreatin indicated improvement in digestibility of proteins after fermentation. The digestibility was further enhanced by steaming.

Adipose tissue is well-recognized as an endocrine organ which secretes a variety of bioactive molecules, including angiotensin II and its precursor angiotensinogen (Agt). There is mounting evidence linking the adipose renin-angiotensin system (RAS) and diet to obesity and obesity-related disorders. However, research addressing dietary regulation and function of adipose RAS is limited, and the specific mechanisms by which PUFAs modulate the endocrine function of adipose tissue remain largely unclear. There are several potential mechanisms that may mediate PUFA effects on Agt, including toll-like receptor signalling, prostaglandins or PPAR-gamma. Thus, we propose to investigate whether PUFAs differentially modulate Agt expression and secretion and to examine possible mechanisms by which PUFA alter Agt expression using the 3T3-L1 cell line. Differentiated 3T3-L1 adipocytes were treated with arachidonic acid (AA), eicosapentaenoic acid (EPA), AA + EPA, or vehicle (C) for 48 hours. Results showed a significant increase in intracellular Agt protein following treatment with PUFAs. Agt secretion, however, was only increased by AA. Interestingly, there is a dose-dependent decrease in Agt protein levels by EPA suggesting that a minimum concentration of n-3 PUFAs is required to elicit an Agt response. Agt mRNA levels were measured by RT-PCR and results showed a significant increase in Agt mRNA in response to treatment with AA but not EPA. These findings suggest that Agt regulation by PUFAs is complex and occurs both post-transcriptionally and post-translationally. Changes in mRNA stability may account for the observed effects of PUFAs. Adipocytes were treated with the transcriptional inhibitor actinomycin D (Act D) and Agt mRNA expression was measured over time. Total RNA was also measured at each time point to ensure that Act D treatment was effectively decreasing transcription. Agt mRNA expression was not significantly altered by treatment with EPA while treatment with AA increased Agt mRNA levels. These results suggest that Agt mRNA stability is differentially increased by n-6 but not n-3 PUFAs. Although there are clear effects of AA on Agt secretion and mRNA stability, the signaling pathways mediating this response remain to be determined, and additional studies are necessary to further dissect the underlying mechanisms of this regulation.

Schizophrenia is one of the most severe mental disorders. Heredity is accepted as a major causative factor. To find molecular mechanisms behind schizophrenia, patient materials with reliable and valid diagnoses must be identified. In order to compare schizophrenia diagnostic procedures for reliability, validity and suitability for genetic studies by evaluation of record information, interview and register diagnostic data and to examine patient materials for linkage or association with molecular genetic markers three patient materials were recruited: sporadic cases, a large pedigree and sib-pairs. Schizophrenia diagnoses based on patient records only, showed good to excellent agreement with diagnoses based on both records and interviews. Register diagnoses generally displayed poor agreement with research diagnoses, but in 94% of patients sometimes registered as schizophrenic psychoses a research diagnosis of these disorders was certified. In the pedigree, analysis suggested linkage to chr 6p23 in a single branch of the pedigree, and a genome scan indicated linkage to the 6q25 region. A genome scan analysis of the sib-pair material was suggestive of linkage to chr 10q25.3-q26.3. In the case-control sample and a meta-analysis there was an association between a dopamine D2 receptor polymorphism (Ser311Cys), on chr 11q22-23, and the disorder. Brain-derived neurotrophic factor gene variants (chr 11p13) were also analysed without any robust significant findings. For patients in long-term treatment for schizophrenia in Sweden, psychiatric record reviews should be valid, reliable and sufficient for assessment of lifetime research diagnosis. Swedish register diagnosis of schizophrenic psychoses has a high positive predictive power in relation to corresponding research diagnoses. For future Swedish studies focusing on a broad definition of schizophrenia, it is sufficient to rely on the register diagnoses of schizophrenic psychosis. There is no major vulnerability gene or locus that is common to the majority of patients with schizophrenia, indicating genetic heterogeneity.

Adipose tissue is well-recognized as an endocrine organ which secretes a variety of bioactive molecules, including angiotensin II and its precursor angiotensinogen (Agt). There is mounting evidence linking the adipose renin-angiotensin system (RAS) and diet to obesity and obesity-related disorders. However, research addressing dietary regulation and function of adipose RAS is limited, and the specific mechanisms by which PUFAs modulate the endocrine function of adipose tissue remain largely unclear. There are several potential mechanisms that may mediate PUFA effects on Agt, including toll-like receptor signalling, prostaglandins or PPAR-gamma. Thus, we propose to investigate whether PUFAs differentially modulate Agt expression and secretion and to examine possible mechanisms by which PUFA alter Agt expression using the 3T3-L1 cell line.Differentiated 3T3-L1 adipocytes were treated with arachidonic acid (AA), eicosapentaenoic acid (EPA), AA + EPA, or vehicle (C) for 48 hours. Results showed a significant increase in intracellular Agt protein following treatment with PUFAs. Agt secretion, however, was only increased by AA. Interestingly, there is a dose-dependent decrease in Agt protein levels by EPA suggesting that a minimum concentration of n-3 PUFAs is required to elicit an Agt response. Agt mRNA levels were measured by RT-PCR and results showed a significant increase in Agt mRNA in response to treatment with AA but not EPA. These findings suggest that Agt regulation by PUFAs is complex and occurs both post-transcriptionally and post-translationally.Changes in mRNA stability may account for the observed effects of PUFAs. Adipocytes were treated with the transcriptional inhibitor actinomycin D (Act D) and Agt mRNA expression was measured over time. Total RNA was also measured at each time point to ensure that Act D treatment was effectively decreasing transcription. Agt mRNA expression was not significantly altered by treatment with EPA while treatment with AA increased Agt mRNA levels. These results suggest that Agt mRNA stability is differentially increased by n-6 but not n-3 PUFAs. Although there are clear effects of AA on Agt secretion and mRNA stability, the signaling pathways mediating this response remain to be determined, and additional studies are necessary to further dissect the underlying mechanisms of this regulation.

The small intestine is the main organ for food digestion and nutrient absorption. It is constantly exposed to antigen and immunomodulatory agents from diet and commensal microbiota. Thus, the intestine is the largest compartment of the immune system in the body. Peanuts and many other allergen resources contain triglycerides, which may affect the antigen absorption through the intestine, but their effects on sensitization and anaphylaxis are unknown. We found that medium chain triglycerides (MCT) promoted antigen absorption into Peyer’s Patches, rather than into the blood directly. Both gavage and feeding of MCT plus peanut protein induced spontaneous allergic sensitization. MCT-sensitized mice experienced the IgG-dependent anaphylaxis from systemic challenges and the IgE-dependent anaphylaxis from oral challenges. Furthermore, MCT alone had direct pharmaceutical effect on enterocytes, like stimulating Jejunal-epithelial Th2 cytokine responses compared with what was seen in the long chain triglycerides (LCT) treated group. Moreover, the oral challenges conducted with peanut protein in MCT significantly exacerbated anaphylaxis compared with the LCT challenges. The intestine also plays an important role in whole body cholesterol homeostasis due to its exclusive function in cholesterol absorption. The researchers found that the intestine function in cholesterol secretion and elimination, but it has not been proven directly until recently. This pathway that facilitate the cholesterol secretions through intestine was named the Transintestinal Cholesterol Efflux (TICE) and has not been well studied yet. To find the possible transporter candidates involved in TICE, we compared both biliary and intestinal cholesterol excretion rates in wild-type (WT) and G5G8 deficient (KO) mice of both sexes. All mice were maintained on a plant-sterol free diet beginning at weaning to prevent the development of secondary phenotypes associated with Sitosterolemia. We found that WT mice had higher biliary cholesterol excretion rates compared to their G5G8 KO littermates as previously reported. However, this difference is significantly greater in females compared to males. Interestingly, intestinal cholesterol excretions increased in female KO mice compared to their WT littermates, a difference not observed in males. This data suggests a sexually dimorphic adaptive mechanism to maintain cholesterol elimination in the absence of G5G8. Whereas male mice maintain a greater level of biliary output in the absence of G5G8, female mice upregulate an alternate intestinal elimination route. To determine the origin of intestinally secreted cholesterol, we compared both hepatobiliary and intestinal cholesterol secretion rates in male wild-type (WT) and CETP transgenic (CETP TG) mice at the age of 12 weeks. Cholesteryl ester transfer protein (CETP) facilitates the transport of cholesteryl esters and triglycerides between lipoproteins in plasma and alters the lipoprotein distribution of plasma cholesterol. We found that WT and CETP TG mice did not differ in either biliary or intestinal cholesterol secretion rates when maintained on a standard chow diet. However, CETP TG mice showed increased biliary cholesterol secretion rates and decreased intestinal cholesterol secretion rates compared to the WT group in response to a Western diet. We next determined the effect of CETP on the delivery of radiolabeled HDL-cholesterol ester to bile and intestinal lumen. Unlike bulk cholesterol secretions, HDL-derived cholesterol esters were preferentially delivered to the intestine in CETP TG mice. This data suggests that CETP alter the routes of total and HDL cholesterol elimination from the body in mice.

Cardiovascular disease remains the leading cause of death in Western societies. Endothelial dysfunction is one of the initiating steps in the development of atherosclerosis. While there is a strong correlation with a person’s genetics, lifestyle factors including smoking, physical activity, and diet can significantly increase a person’s susceptibility to the development of atherosclerosis. In addition to these lifestyle factors, there is a strong body of evidence linking exposure to environmental pollutants including persistent organic pollutants such as polychlorinated biphenyls to increased cardiovascular disease and mortality. It has been well-established that exercise protects against cardiovascular disease, but whether exercise can modulate PCB-induced cardiovascular inflammation and dysfunction is unknown. To investigate the effects of exercise on PCB-induced cardiovascular disease, two murine models of atherosclerosis, the ApoE-/- and the LDLr-/- mouse were utilized. Risk factors for cardiovascular disease including adiposity, glucose intolerance, hyperlipidemia, hypertension, oxidative stress, and inflammation, were assessed in these two models as well as mean atherosclerotic lesion size. Exercise positively modulates several risk factors associated with cardiovascular disease including hypertension, hyperlipidemia, adiposity and obesity, systemic levels of oxidative stress, inflammation, and glucose tolerance. Exercise significantly reduced mean lesion size in vehicle-treated animals. To assess the mechanism of protection of exercise in chapter 4, vascular reactivity studies were performed to measure endothelial function after exposure to PCB 77. Exercise prevented PCB-impaired endothelial function implicating the role of superoxide as a cause of impairment. Exercise upregulated phase II antioxidant enzymes. The work in this dissertation demonstrates several protective properties of exercise against PCB-induced cardiovascular disease; however, additional studies are needed to determine if exercise enhances metabolism and excretion of these environmental pollutants.

Made available in DSpace on 2015-04-17T14:55:18Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3272865 bytes, checksum: 5113c553db939bb931f3c23e02c51ae2 (MD5) Previous issue date: 2011-09-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Soil saliniy is a major factor in reducing plant growth and, consequently, their agricultural productivity. Most of the studies in the literature presents a strategy for reducing the effects of improving salt tolerance in plants for the production of osmolytes and stress proteins, and there are few studies aimed at increasing salt tolerance by restoring ionic homeostasis. This work was proposed to understand the physiological mechanisms and structural related with adjustment in the halophyle Halococcus morrhuae. This showed significant growth in the medium to halobacterias, that are organized in the form of sarcina of different sizes. The results achieved in attempts to characterize the lipid membrane halobactéria according to the values of retention time corresponding to phosphatidic acid (PA) and phosphatidylglycerol (C20-C20) (PG), and glycolipid known as Diglicosilarqueol (DGA-1 ). Additionally, patterns of breaks fatty acids extracted from Halococcus morrhuae showed a significant similarity when compared with the fatty acids found in organisms of the domain Eucarya and Bacteria. The genes of the antiport Na + / Ca2+ and Na + / H+ proved functional after cloning in Escherichia coli DH5α in solid and liquid medium. The antiport Na+ / H+ activity showed a statistically more significant when compared to the growth of clones containing the antiport Na+/Ca2+, making it interesting for the construction of transgenic plants tolerant to salinity. These database contribute to understanding the physiology of Halococcus morrhuae for future biotechnological applications, among them the inclusion of these genes in plants to assess the functionality in eukaryotic models. / A salinização do solo é um dos principais fatores para redução do crescimento das plantas e, consequentemente, da sua produtividade agrícola. A maioria dos estudos presentes na literatura apresenta como estratégia para redução dos seus efeitos o melhoramento da tolerância a sal pelas plantas pela produção de osmólitos e proteínas de estresse, existindo poucos estudos visando o aumento da tolerância ao sal pelo restabelecimento da homeostase iônica. Neste trabalho foi proposto compreender mecanismos fisiológicos e estruturais relacionados com a adaptação à halofilia em Halococcus morrhuae. O crescimento de H. morrhuae foi significativo no meio para halobactérias, organiza-se na forma de sarcinas de diferentes tamanhos ao longo do tempo. Os resultados alcançados na tentativa de caracterização de lipídios da membrana da halobactéria, de acordo com os valores de tempo de retenção, correspondem ao ácido fosfatídico (PA) e fosfatidilglicerol (C20-C20)(PG), e ao glicolipídio conhecido como diglicosilarqueol (DGA-1). Adicionalmente, os padrões de quebras dos ácidos graxos extraídos de Halococcus morrhuae apresentaram uma similaridade significativa quando comparada com a dos ácidos graxos encontrados em organismos do domínio Eucarya e Bacteria. Os genes dos antiportes Na+/Ca+2 e Na+/H+ aumetaram a tolerância ao sal, mostrando-se funcionais após espressão em Escherichia coli DH5α em meio sólido e em meio líquido. O antiporte Na+/H+ apresentou uma atividade estatisticamente mais significativa (p<0,03) quando comparada ao crescimento dos clones contendo o antiporte Na+/Ca2 (p<0,05). Entretanto, ambos os antiportes são candidatos interessantes para a construção de plantas geneticamente modificadas tolerantes a salinidade. Estes dados também contribuem para o entendimento da fisiologia de H. morrhuae para futuras aplicações biotecnológicas, dentre elas a inserção desses genes em plantas para avaliar a funcionalidade em modelos eucarióticos.

Made available in DSpace on 2015-04-01T14:16:00Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3272893 bytes, checksum: 54fd73a37837f8d64e245874e3ff9a98 (MD5) Previous issue date: 2011-09-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Soil saliniy is a major factor in reducing plant growth and, consequently, their agricultural productivity. Most of the studies in the literature presents a strategy for reducing the effects of improving salt tolerance in plants for the production of osmolytes and stress proteins, and there are few studies aimed at increasing salt tolerance by restoring ionic homeostasis. This work was proposed to understand the physiological mechanisms and structural related with adjustment in the halophyle Halococcus morrhuae. This showed significant growth in the medium to halobacterias, that are organized in the form of sarcina of different sizes. The results achieved in attempts to characterize the lipid membrane halobactéria according to the values of retention time corresponding to phosphatidic acid (PA) and phosphatidylglycerol (C20-C20) (PG), and glycolipid known as Diglicosilarqueol (DGA-1 ). Additionally, patterns of breaks fatty acids extracted from Halococcus morrhuae showed a significant similarity when compared with the fatty acids found in organisms of the domain Eucarya and Bacteria. The genes of the antiport Na + / Ca2+ and Na + / H+ proved functional after cloning in Escherichia coli DH5α in solid and liquid medium. The antiport Na+ / H+ activity showed a statistically more significant when compared to the growth of clones containing the antiport Na+/Ca2+, making it interesting for the construction of transgenic plants tolerant to salinity. These database contribute to understanding the physiology of Halococcus morrhuae for future biotechnological applications, among them the inclusion of these genes in plants to assess the functionality in eukaryotic models. / A salinização do solo é um dos principais fatores para redução do crescimento das plantas e, consequentemente, da sua produtividade agrícola. A maioria dos estudos presentes na literatura apresenta como estratégia para redução dos seus efeitos o melhoramento da tolerância a sal pelas plantas pela produção de osmólitos e proteínas de estresse, existindo poucos estudos visando o aumento da tolerância ao sal pelo restabelecimento da homeostase iônica. Neste trabalho foi proposto compreender mecanismos fisiológicos e estruturais relacionados com a adaptação à halofilia em Halococcus morrhuae. O crescimento de H. morrhuae foi significativo no meio para halobactérias, organiza-se na forma de sarcinas de diferentes tamanhos ao longo do tempo. Os resultados alcançados na tentativa de caracterização de lipídios da membrana da halobactéria, de acordo com os valores de tempo de retenção, correspondem ao ácido fosfatídico (PA) e fosfatidilglicerol (C20-C20)(PG), e ao glicolipídio conhecido como diglicosilarqueol (DGA-1). Adicionalmente, os padrões de quebras dos ácidos graxos extraídos de Halococcus morrhuae apresentaram uma similaridade significativa quando comparada com a dos ácidos graxos encontrados em organismos do domínio Eucarya e Bacteria. Os genes dos antiportes Na+/Ca+2 e Na+/H+ aumetaram a tolerância ao sal, mostrando-se funcionais após espressão em Escherichia coli DH5α em meio sólido e em meio líquido. O antiporte Na+/H+ apresentou uma atividade estatisticamente mais significativa (p<0,03) quando comparada ao crescimento dos clones contendo o antiporte Na+/Ca2 (p<0,05). Entretanto, ambos os antiportes são candidatos interessantes para a construção de plantas geneticamente modificadas tolerantes a salinidade. Estes dados também contribuem para o entendimento da fisiologia de H. morrhuae para futuras aplicações biotecnológicas, dentre elas a inserção desses genes em plantas para avaliar a funcionalidade em modelos eucarióticos.