O gênero Capsicum compreende mais de 200 espécies de pimentas e pimentões e diversas atividades farmacológicas são descritas na literatura. Estudos prévios do nosso grupo de pesquisa têm explorado diferentes atividades biológicas de Capsicum baccatum var. Pendulum (Solanaceae), como antioxidante, antiinflamatória, hipoglicemiante e antidislipidêmica. A prevalência da síndrome metabólica, caracterizada por fatores como obesidade, resistência à insulina, dislipidemia e hipertensão, tem atingido proporções epidêmicas e acomete cerca de 20% da população mundial. Muitas vezes, apenas a mudança no estilo de vida não é eficaz para impedir a progressão da doença, sendo necessário aderir ao tratamento farmacológico. No entanto, não existem fármacos específicos para o tratamento da síndrome, levando à utilização de polifarmácia, que gera dificuldades de adesão ao tratamento e risco de interações medicamentosas. O principal objetivo desse estudo foi avaliar o efeito do extrato butanólico (BUT) e da fração ativa (AF) dos frutos de C. baccatum em um modelo de síndrome metabólica induzido por dieta em camundongos C57BL/6. Inicialmente apresentamos uma revisão de estudos publicados, que utilizam diferentes tipos de dietas hipercalóricas para induzir a síndrome metabólica em roedores. Ao compararmos esses estudos, observamos a forte tendência à utilização de camundongos da linhagem C57BL/6 machos e jovens, respondendo de forma satisfatória à indução da síndrome metabólica. Após, apresentamos o efeito do BUT e AF, nas doses de 200 e 50 mg/kg, respectivamente, sobre os parâmetros glicídicos e lipídicos em modelo de síndrome metabólica em camundongos C57BL/6 submetidos a uma dieta hiperpalatável (HPD) durante 120 dias. Nesse estudo foi visto que o tratamento com C. baccatum via oral, preveniu o ganho de peso, o desenvolvimento de um perfil de intolerância à glicose, o acúmulo de gordura na região abdominal e a elevação nos níveis de insulina e leptina plasmáticas quando comparados ao grupo HPD. Além disso, também preveniu o acúmulo de colesterol e triglicerídeos no fígado e preservou a morfologia do tecido hepático. Assim, C. baccatum se mostrou promissor, prevenindo alterações dos diversos componentes importantes da síndrome metabólica. / The genus Capsicum comprises more than 200 species of peppers and a range of pharmacological activities are described in the literature. Previous studies of our research group have explored several activities for Capsicum baccatum var. pendulum (Solanaceae) species, as antioxidant, anti-inflammatory, hypoglycemic and antidyslipidemic, since the studies with this species are scarce. The prevalence of metabolic syndrome, characterized by the association of factors such as obesity, insulin resistance, dyslipidemia and hypertension, is reaching epidemic proportions worldwide, affecting approximately 20% of world's population. Often, only lifestyle change is not effective in preventing the progression of the disease, being necessary adherence to pharmacological treatment. However, there are no specific drugs for the treatment of the syndrome, leading to the use of polypharmacy, which generates an increase in public spending, difficulties of adherence to treatment, and enhanced risk to drug interactions. The main objective of this study was to evaluate the effect of butanolic extract (BUT) and the active fraction (AF) of C. baccatum on a model of diet-induced metabolic syndrome in C57BL/6 mice. We first present a review of published studies, which use different types of hypercaloric diets to induce metabolic syndrome in rodents. When comparing these studies, we observed a strong trend towards the use of male and young C57BL/6 mice, responding satisfactorily to the metabolic syndrome induction through hypercaloric diets. After, we present the effect of BUT and AF, on glucose and lipid parameters in a model of metabolic syndrome in C57BL/6 mice submitted to a hyperpalatable diet (HPD) for 120 days. In this study, it was seen that oral C. baccatum treatment prevented weight gain, development of a glucose intolerance profile, accumulation of fat in the abdominal area and the increase in plasma insulin and leptin levels when compared to group HPD. Also, it prevented the accumulation of cholesterol and triglycerides in the liver, and preserved the liver tissue morphology. Thus, these results showed the effects of C. baccatum, especially AF, preventing changes in several important components of the metabolic syndrome.

Cryptosporidium parvum is a coccidian protozoan that colonizes epithelial cells lining respiratory and digestive tracts of animals and humans. Cryptosporidiosis is a well-recognized zoonotic disease infecting primarily neonates and immunocompromised hosts, including human immunodeficiency virus-infected patients. Clinical disease is manifested as a chronic diarrheal illness that is self-limiting in immunocompetent hosts and prolonged and often life-threatening in hosts with compromised immune systems. The lack of a suitable small animal model for screening anti-cryptosporidial drugs and for examining the pathogenicity and immunobiology of chronic cryptosporidosis was the impetus for this research effort. The objectives of the present study were three-fold: to characterize chronic Cryptosporidium parvum infections in dexamethasone-immunosuppressed mice; evaluate the effects of Cryptosporidium parvum and dexamethasone on B and T lymphocyte proliferation; and determine the effects of the immunomodulator dehydroepiandrosterone on oocyst shedding intensities of mice infected with Cryptosporidium parvum Adult C57BL/6N mice were immunosuppressed with the synthetic glucocorticoid dexamethasone, then infected with Cryptosporidium parvum (106 oocysts/mouse) and investigated for their ability to sustain a four-month chronic infection. Dexamethasone was administered intraperitoneally (125μ/mouse/day) or orally (8μ/ml) in the drinking water ad libitum. Infection chronicity was characterized by evaluating mouse mortality, oocyst excretion in the feces, tissue distribution of the parasite, and the parasite-induced pathology. A progressive infection with Cryptosporidium parvum occurred in mice immunosuppressed intraperitoneally and orally as long as dexamethsone was administered. Mice receiving dexamethasone given intraperitoneally had a shorter prepatent period and a more consistent, although cyclic, oocyst shedding pattern when compared with mice given dexamethasone orally. Mice given dexamethasone orally exhibited a delayed prepatent period, with a steady increase in oocyst shedding. All mice receiving dexamethasone orally died within three months following oocyst inoculation. Clinical signs included dehydration, icterus, and reduction in spleen and body weights. Clinical signs were more abrupt in mice receiving oral dexamethasone. Parasite colonization involved the entire intestinal tract, including the pyloric ring and Peyer's patches, but was the heaviest in the terminal ileum. Parasites were present in the lungs, gallbladder, and pancreatic ducts. Pathologic abnormalities were isolated to the terminal small intestine and included blunting and fusion of intestinal villi and crypt hyperplasia. Cryptosporidium parvum and dexamethasone administered in vivo reduced B and T lymphocyte responses to the mitogens lipopolysaccharide and concanavalin A. Dehydroepiandrosterone and dehydroepiandrosterone-sulfate resulted in no significant reductions in cryptosporidial activity as determined by oocyst shedding in the feces.

Voluntary wheel running is rewarding and believed to activate the same brain reward system as in alcohol and drug addiction. Brain-derived neurotrophic factor (BDNF), a well-known growth factor widely expressed in the brain, is modulated by both voluntary exercise and alcohol consumption. The aim of this study was to evaluate how voluntary exercise affects the expression levels of BDNF and its receptor TrkB in brain regions involved in positive and negative reinforcement. Additionally we wanted to evaluate the effect it may have on alcohol drinking behaviors in C57BL/6 mice, a mouse model which are naturally prone for engaging in voluntary exercise and voluntary alcohol consumption. We found a small upregulation in DG and CA1 after three weeks of exercise, confirming findings by others, and a significant 3-fold downregulation of BDNF in NAc after both three weeks of exercise and exercise followed by a five week period of either ethanol intake or not. Interestingly, we here show a significant 100-fold increase in BDNF after exercise and a 120-fold increase after both exercise and alcohol consumption in amygdala, a region involved in regulation of anxiety-related behavior and negative reinforcement. Additionally a slightly lower 10-fold increase in BDNF was seen after exercise and a 15-fold increase after exercise followed by ethanol in prefrontal cortex, a structure contributing to reward-related behavior. Behaviorally, we could not either directly following exercise or at five weeks post-exercise detect any significant effect of wheel-running on depression-related behavior. However, we did find that exercise significantly increased the alcohol intake.

<p>Pathologies of sexual development are common in humans and reflect the precarious processes of sex determination and sexual differentiation. The gonad forms as a bipotential organ, and recent results from the Capel lab revealed that it is initially balanced between testis and ovarian fates by opposing and antagonistic signaling networks. In XY embryos, this balance is disrupted by the transient expression of the Y-linked gene, Sry, which activates genes that promote the testis pathway and oppose the ovarian pathway. While the roles of a few genes have been defined by mutation, current evidence suggests that the interactions of many genes and signaling pathways are involved in the establishment of sexual fate. For example, most cases of disorders of sexual development (DSDs) are unexplained by mutations in known sex determination genes. In addition, recent microarray studies in the mouse revealed that nearly half the transcriptome is expressed in the gonad at the time of sex determination (Embryonic day 11.5, or E11.5), and as many as 1,500 genes are expressed in a sexually dimorphic pattern at this early stage. Thus the sexual fate decision in the developing gonad likely depends on a complex network of interacting factors that converge on a critical threshold. </p><p>To begin to elucidate the transcription network topology underlying sex determination, we exploited two inbred mouse strains with well-characterized differences in sex reversal. The common inbred strain C57BL/6J (B6) is uniquely sensitive to XY male-to-female sex reversal in response to a number of genetic perturbations, while other strains, including 129S1/SvImJ (129S1) and DBA/2J (D2) are resistant to sex reversal. We hypothesized that these strain differences in gonad phenotype likely result from underlying expression differences in the gonad at the critical timepoint of E11.5. Using microarrays, we identified significant, reproducible differences in the transcriptome of the E11.5 XY gonad between B6 and 129S1 indicating that the reported sensitivity of B6 to sex reversal is consistent with a higher expression of a female-like transcriptome in B6 XY gonads. Surprisingly, a well-characterized master regulator of the testis pathway, Sox9, was found to be upregulated in the sensitive B6 background, which may serve as a compensatory mechanism to counteract the female-leaning transcriptome and activate the testis pathway in wild type B6 XY gonads.</p><p>We extended our expression analysis to a large set of F2 XY gonads from B6 and 129S1 intercrosses. From each pair of gonads, we analyzed the expression of 56 sex-associated genes by nanoliter-scale quantitative RT-PCR (qRT-PCR). The expression levels of most genes were highly variable across the F2 population, yet strong correlations among genes emerged. We employed a First-Order Conditional Independence (FOCI) algorithm to estimate the F2 coexpression network. From this unbiased analysis of XY expression data, we uncovered two subnetworks consisting of primarily male and female genes. Furthermore, we predicted roles for genes of unknown function based on their connectivity and position within the network. </p><p>To identify the genes responsible for these strain expression differences, we genotyped each F2 embryo at 128 single nucleotide polymorphisms (SNPs) located evenly throughout the 19 autosomes and X chromosome. We then employed linkage analysis to detect autosomal regions that control the expression of one or more of the 56 genes in the F2 population. These regions are termed expression quantitative trait loci, or eQTLs. We identified eQTLs for many sex-related genes, including Sry and Sox9, the key regulators of male sex determination. In addition, we identified multiple prominent trans-band eQTLs that controlled the expression of many genes. My work represents the first eQTL analysis of a developing vertebrate organ, the mouse gonad. This systems-level approach revealed the complex transcription architecture underlying sex determination, and provides a mechanistic explanation for sensitivity to sex reversal seen in some individuals.</p> / Dissertation

Thesis (M.Sc.). / Written for the Dept. of Microbiology and Immunology. Title from title page of PDF (viewed 2008/07/30). Includes bibliographical references.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2009. / Title from first page of PDF file (viewed on June 8, 2009). Includes bibliographical references.

Thesis (Ph.D.). / Written for the Dept. of Biomedical Engineering. Title from title page of PDF (viewed 2009/06/11). Includes bibliographical references.

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Made available in DSpace on 2015-12-06T23:46:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2014 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Atualmente cerca de 13% da populacao mundial e afetada por algum grau de doenca renal cronica, com um percentual de 45% de mortes atribuidas a desordens de carater fibrotico. Neste contexto, novas formas terapeuticas de tratamento se tornam imprescindiveis, como por exemplo, uso de terapia celular com celulas-tronco mesenquimais. Essas celulas podem ser obtidas do tecido-adiposo (CTAd) tornando-se possiveis terapias devido a sua facil obtencao e propriedades pro-regenerativas, principalmente as exercidas por acoes paracrinas. Portanto, neste presente trabalho analisamos o papel da CTAd na reducao da progressao da fibrose renal em dois modelos experimentais: 1) de isquemia e reperfusao renal unilateral severa (IR) e 2) de nefrite tubulo-intersticial (NTI). Inicialmente, as CTAd foram isoladas de camundongos C57BL/6j, e foram caracterizadas fenotipicamente e funcionalmente. Apos sua caracterizacao, as CTAds foram administradas por via intraperitoneal (2.105 celulas/animal) em diferentes tempos apos insulto inicial, e os animais foram avaliados 24 horas, 6 e 10 semanas apos a IR e 10 dias apos a inducao da NTI. No modelo de IR, no tempo de 24 horas, os animais tratados com CTAd apresentaram reduzida disfuncao renal e tubular e um aumento do processo regenerativo. Adicionalmente, a expressao renal de transcritos pro-inflamatorios (IL-6 e TNF) foi diminuida, enquanto que o nivel de moleculas antiinflamatorias e citoprotetoras (IL-4, IL-10 e HO-1) foram aumentadas. Apesar da melhora funcional, as CTAd nao foram observadas nos rins dos animais transplantados. Como esperado, apos 6 semanas de IR, os rins dos animais nao tratados retrairam, enquanto os rins dos animais tratados com CTAd permaneceram integros com menor deposicao de fibras de colageno do tipo 1 (Col-1) e menos expressao do marcador de fibrose FSP-1. A protecao renal observada em animais tratados com CTAd foi acompanhada por uma reducao nos niveis sericos de citocinas inflamatorias como TNF-&#945;, KC, RANTES e IL-1a. No modelo de NTI o mesmo padrao foi observado, ou seja, os rins dos animais tratados apresentaram menor disfuncao renal com reduzida expressao de moleculas pro-inflamatorias (TNF-a e IL-6) e diminuicao da area fibrotica observada principalmente pela coloracao de Picrosirius e imunohistoquimica para marcadores pro-fibroticos como FSP-1 e Col-1. Surpreendentemente, quando avaliamos o tratamento das CTAd com 6 semanas de IR, periodo em que os animais ja apresentavam fibrose, foi constatado uma melhora nos parametros funcionais e menos fibrose e reducao da expressao de RNAm de Col-1 e vimentina, alem de menor expressao de FSP-1. Conclusivamente, nossos resultados demonstram que a terapia com CTAd pode afetar a progressao da fibrose renal, pela modulacao da resposta inflamatoria inicial. Acreditamos que essa protecao possa estar relacionada a modulacao dos mecanismos que governam a transicao epitelial/endotelio-mesenquimal. Esperamos que esse trabalho possa contribuir para a identificacao de potenciais vias de lesao ou de regeneracao, facilitando a sua traducao mais simples e rapida para a pratica clinica / BV UNIFESP: Teses e dissertações

Made available in DSpace on 2015-07-22T20:50:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-31 / O aumento da neurotrasmissão dopaminérgica tem sido ligado à euforia produzida pela cocaína. Estudos pré-clínicos demonstraram que a modificação da composição de ácidos gráxos poli-insadurados (PUFAs) na dieta pode influenciar o nível de neurotransmissão. O objetivo do estudo foi verificar se a administração de PUFAs provoca alteração na preferência condicionada por lugar por cocaína, correlacionando os dados comportamentais com os níveis plasmáticos de PUFAs. Foram utilizados 5 grupos de camundongos c57BI/6J suplementados com óleo de linhaça ou solução controle por 19 dias e condicionados com cocaína, embora tenham sido observadas alerações significativas dos níveis de ácido eicosapentaenóico (EPA) e ácido linoléico entre os grupos. Doses maiores de PUFAS são necessárias para que ocorra mudança no nível plasmático de EPA em camundongos condicionados com cocaína. / TEDE / BV UNIFESP: Teses e dissertações