Made available in DSpace on 2014-06-11T19:23:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-19Bitstream added on 2014-06-13T18:50:47Z : No. of bitstreams: 1 carvalho_fg_me_arafcf.pdf: 662906 bytes, checksum: b01eca6154c2a1b53dcdc4ffa1a8d2c8 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / Mulheres pós-menopausadas estão mais susceptíveis a problemas de saúde relacionados hipoestrogenismo, o que favorece ao processo de estresse oxidativo, inflamatório e ao desenvolvimento de doenças crônicas. Visto que a inclusão de cereais integrais na alimentação veicula componentes bioativos de efeito antioxidante e hipolipemiante, como as lignanas, o objetivo do presente estudo foi avaliar os efeitos hipolipemiantes, inflamatórios e antioxidantes de lignanas provenientes da ingestão de quinoa (Chenopodium quinoa) em um grupo de mulheres pós-menopausadas. Foi realizado um estudo prospectivo, randomizado, duplo cego e controlado por placebo, no qual participaram 35 mulheres que foram submetidas ao consumo diário de 25 gramas de quinoa em flocos ou placebo, no período de 4 semanas consecutivas. No início e ao final do tratamento, após as quatro semanas, foram realizadas avaliações antropométricas: peso corporal, estatura e circunferência da cintura; e coleta de sangue para a quantificação de glicose, colesterol total, LDL-C, HDL-C, triglicerídeos, marcadores de estresse (GSH e TBARS), Vitamina E, marcadores inflamatórios (IL-6 e TNF-α) e enterolignanas (END e ENL); e urina de 24 horas para quantificação de enterolignanas. Os resultados obtidos foram analisados em dois diferentes estudos os quais foram escritos na forma de artigo científico, sendo que o primeiro artigo abordou o efeito do consumo de quinoa sobre as concentrações de glicose, colesterol total e frações e de marcadores de estresse oxidativo, e o segundo artigo abordou o efeito das enterolignas sobre os marcadores inflamatórios em um grupo de mulheres pós-menopausadas. Ao comparar as dosagens no início e ao final do experimento, o presente estudo mostrou um possível efeito benéfico proveniente da ingestão do cereal quinoa, pois foram constatadas reduções significativas nas concentrações... / Postmenopausal women are more susceptible to health problems related to declining estrogen concentrations, which favor the oxidative stress and inflammatory process and the development of chronic diseases. Since daily consumption of grains involves bioactive components with an antioxidant and hypolipidemic effects, like lignans, the aim of the present study was to investigate the effect of quinoa consumption on the concentrations of glucose, total cholesterol and fractions, oxidative stress and inflammatory markers in a group of postmenopausal women. A prospective, randomized, double-blind and placebo-controlled study has been conducted on 35 women who had to consume 25 grams/day of quinoa flakes or placebo, over a period of 4 consecutive weeks. At the beginning and at the end of the intervention, after four weeks, anthropometric assessment was performed by body weight, height and abdominal circumference; and blood was collected for the determination of glucose, total cholesterol and fractions, oxidative stress and inflammatory markers, vitamin E and enterolignans, and 24-h urine was obtained for the determination of enterolignans. The results were analyzed in two different studies which were written in the form of a scientific paper, the first one emphasizes the effect of quinoa intake on consumption on the concentrations of glucose, total cholesterol and fractions, oxidative stress markers and in the second paper investigated the effect of enterolignans on inflammatory markers in a group of postmenopausal women. Comparing the beginning and the end of the intervention, the present study showed a possible beneficial effect by quinoa intake, because significant reductions were observed in serum triglycerides, TBARS and vitamin E concentrations, and an increase in enterolignan urinary excretion in the groups that consumed quinoa and placebo. A significant reduction of total cholesterol... (Complete abstract click electronic access below)

O consumo de álcool tem sido considerado um problema de saúde pública. Entretanto, também existem evidências de eventuais benefícios do consumo leve a moderado de algumas bebidas alcoólicas, no que se refere a doenças crônicas não transmissíveis. As doenças periodontais, sendo a sexta doença crônica mais prevalente, também têm sido associadas ao consumo de álcool com resultados controversos. Esta tese avaliou, através de três artigos a relação entre consumo de álcool e doenças periodontais. O primeiro artigo de divulgação, tem por objetivo alertar a profissão a respeito do estado atual do conhecimento em relação a associação entre álcool e doenças periodontais. O segundo artigo é um estudo experimental em modelo animal que avaliou o efeito da dependência química álcool sobre desfechos periodontais. Os resultados não mostraram diferenças estatisticamente significativas na perda óssea alveolar e secreção de TNF-α. O terceiro artigo, também em modelo animal, abordou o consumo do vinho tinto e valeu-se de controles total, álcool a 12%, suco de uva e resveratrol. A análise de ocorrência de periodontite espontânea demonstrou que o vinho tinto tem potencial protetor para a perda óssea alveolar e secreção de TNF- α . Os resultados encontrados nessa tese são instigantes e estão em linha com a literatura que demonstra uma relação do tipo curva J entre álcool e doenças periodontais, na qual até um determinado nível, o fator comporta-se como protetor. Assim, essa relação deve ser considerada na abordagem clínica, sem incentivo ao consumo, mas compreendendo até onde este é tolerável e/ou benéfico no que se refere a doenças periodontais. / Alcohol consumption has been considered a public health problem. Otherwise, there is evidence of eventual benefits of a light to moderate consume of some alcoholic beverage, in relation to non-transmissible chronic disease. The periodontal diseases, being the sixth most prevalent chronic disease, also have been associated to alcohol consumption with controversial results. This thesis evaluated, through three articles the relationship between alcohol consumption and periodontal diseases. The first article has the objective of alerting the professionals about the actual knowledge about the relationship between alcohol and periodontal diseases. The second article is an experimental study in animal model which evaluated the alcohol dependence over periodontal outcomes. The results did not show statistically significant differences in alveolar bone loss and TNF- α secretion. The third article, also in animal model, studied the red wine consumption and used total controls, 12% alcohol, grape juice and resveratrol. The analysis of the occurrence of spontaneous periodontitis demonstrated that red wine has a protector potential for alveolar bone loss and TNF- α secretion. The results found in this thesis are curious and they are in line with the literature that demonstrates a relationship like J-curve between alcohol and periodontal diseases, until some level, the factor behave as protector. So, this relation must be considered in clinical approaches, without encouraging the consumption, but understanding how much is tolerable and/or benefit according to periodontal diseases.

O consumo de álcool tem sido considerado um problema de saúde pública. Entretanto, também existem evidências de eventuais benefícios do consumo leve a moderado de algumas bebidas alcoólicas, no que se refere a doenças crônicas não transmissíveis. As doenças periodontais, sendo a sexta doença crônica mais prevalente, também têm sido associadas ao consumo de álcool com resultados controversos. Esta tese avaliou, através de três artigos a relação entre consumo de álcool e doenças periodontais. O primeiro artigo de divulgação, tem por objetivo alertar a profissão a respeito do estado atual do conhecimento em relação a associação entre álcool e doenças periodontais. O segundo artigo é um estudo experimental em modelo animal que avaliou o efeito da dependência química álcool sobre desfechos periodontais. Os resultados não mostraram diferenças estatisticamente significativas na perda óssea alveolar e secreção de TNF-α. O terceiro artigo, também em modelo animal, abordou o consumo do vinho tinto e valeu-se de controles total, álcool a 12%, suco de uva e resveratrol. A análise de ocorrência de periodontite espontânea demonstrou que o vinho tinto tem potencial protetor para a perda óssea alveolar e secreção de TNF- α . Os resultados encontrados nessa tese são instigantes e estão em linha com a literatura que demonstra uma relação do tipo curva J entre álcool e doenças periodontais, na qual até um determinado nível, o fator comporta-se como protetor. Assim, essa relação deve ser considerada na abordagem clínica, sem incentivo ao consumo, mas compreendendo até onde este é tolerável e/ou benéfico no que se refere a doenças periodontais. / Alcohol consumption has been considered a public health problem. Otherwise, there is evidence of eventual benefits of a light to moderate consume of some alcoholic beverage, in relation to non-transmissible chronic disease. The periodontal diseases, being the sixth most prevalent chronic disease, also have been associated to alcohol consumption with controversial results. This thesis evaluated, through three articles the relationship between alcohol consumption and periodontal diseases. The first article has the objective of alerting the professionals about the actual knowledge about the relationship between alcohol and periodontal diseases. The second article is an experimental study in animal model which evaluated the alcohol dependence over periodontal outcomes. The results did not show statistically significant differences in alveolar bone loss and TNF- α secretion. The third article, also in animal model, studied the red wine consumption and used total controls, 12% alcohol, grape juice and resveratrol. The analysis of the occurrence of spontaneous periodontitis demonstrated that red wine has a protector potential for alveolar bone loss and TNF- α secretion. The results found in this thesis are curious and they are in line with the literature that demonstrates a relationship like J-curve between alcohol and periodontal diseases, until some level, the factor behave as protector. So, this relation must be considered in clinical approaches, without encouraging the consumption, but understanding how much is tolerable and/or benefit according to periodontal diseases.

Menopause transition is usually associated with changes in body composition and a decrease in physical activity energy expenditure. Adipose tissue, especially visceral fat, is an important source of inflammatory markers, which contributes to the development of a pro-inflammatory state. Conversely, high levels of physical activity and exercise have an anti-inflammatory effect. One-hundred and two healthy premenopausal women participated in a 5-year longitudinal observational study (MONET: Montreal Ottawa New Emerging Team). The present secondary analyses were performed on 58 participants between the ages of 47 and 54 years with a full set of data.The aim of study was to investigate the impact of menopause transition and physical activity on inflammatory makers. The major finding of the first of 3 studies was that menopausal transition is accompanied by an increase in inflammatory markers, namely ferritin, IL-8, and sTNFR 1 and 2. The increase in IL-8 and sTNFR2 with menopause could be explained, in part, by changes in fat mass and peripheral fat, respectively. During and after menopause, significant bone loss occurs in women due to reduced estrogen production. Estrogen reduction favors bone resorption by regulating the production and activity of inflammatory markers. Therefore we further investigated the association between inflammatory markers and bone mineral density in premenopausal women transitioning to menopause (paper 2). Our results showed no significant association between change in inflammatory markers and change in bone mineral density in women transitioning to menopause. However, in premenopausal women hs-CRP was negatively associated with total, lumbar spine and femoral neck bone mineral density and along with weight and cardiorespiratory fitness may play a role in bone mineral density variation. Baseline level of hs-CRP, Hp, IL-6 and femoral neck bone mineral density along with percent change in physical activity energy expenditure and menopausal status partly explained the individual variation of bone mineral density losses in women transitioning to menopause. Finally, we investigated time spent in the postmenopausal years and the influence of the duration of the postmenopause status on body composition and cardiometabolic risk factors. We indicated that postmenopausal years and years since menopause is associated with decrease in blood glucose and increase in waist circumference, percent fat mass, total cholesterol, and high density lipoprotein. Inflammatory markers including ApoB, ferritin, adiponectin, sCD14 were higher during years after final menstrual period while sTNFR1 and sTNFR2 were higher during the menopause transition and early postmenopausal years.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Methotrexate (MTX) is an antimetabolite drug analogue of folic acid with wide clinical application, used in high doses for the treatment of cancer and in low doses for the treatment of autoimmune diseases such as rheumatoid arthritis and psoriasis. Although effective, the MTX has several side effects. The oxidative stress seems to be involved with the toxicity caused by the MTX in various organs. Despite the effect on the imbalance of oxidative metabolism, the influence of polymorphisms of antioxidant enzymes on MTX toxicity is not well studied. In this context, the present study aimed to examine whether the Ala16Val polymorphism of the antioxidant enzyme superoxide dismutase manganese dependent (SOD2), which affects the efficiency of the detox enzyme, could have an effect on the cytotoxic response to MTX. For this, an in vitro study using peripheral blood mononuclear cells (PBMC) obtained from healthy donors harboring different genotypes of polymorphism Ala16Val-SOD (= genotypes AA, VV and AV) was performed. Once obtained, PBMCs were treated with MTX at concentrations of 10 and 100 μM for 24 and 72 hours and analyzed for the effect on viability, modulation of the oxidative metabolism-inflammatory and apoptotic. PBMC-AA which have a naturally SOD2 30 to 40% more efficient than the PBMC- VV, showed more resistance to treatment with MTX compared to PBMC-AV/VV assessed. As production levels of EROS and lipid peroxidation significantly increased in cells exposed to MTX, regardless of genotype, however, increased levels of protein carbonylation were observed only in PBMC-AV/VV. The PBMC-AA demonstrated decreased activity of SOD2 and the levels of glutathione peroxidase with PBMC-AA were higher. The levels of caspase-8 and -3 were increased in PBMC exposed to MTX, but the modulation of these genes, as well as Bax and Bcl-2 genes involved in apoptotic route, was genotype dependent. The MTX was able to raise the levels of inflammatory cytokines and decrease the level of anti-inflammatory cytokine IL-10, regardless of genotype. The results suggest that Ala16Val-SOD2 polymorphism is capable of modulating the cytotoxic response of PBMC to the MTX. / O Metotrexato (MTX) é um fármaco antimetabólito análogo do ácido fólico, com vasta aplicação clinica, utilizado em doses elevadas no tratamento de neoplasias e em baixas doses para o tratamento de doenças autoimunes, como a artrite reumatoide e a psoríase. Apesar de efetivo, o MTX possui diversos efeitos colaterais. Assim, o estresse oxidativo parece estar envolvido com a toxicidade causada pelo MTX a diversos órgãos. Apesar do efeito no desequilíbrio do metabolismo oxidativo, a influência de polimorfismos de enzimas antioxidantes sobre a toxicidade ao MTX ainda não é bem estudada. Nesse contexto, o presente estudo teve como objetivo analisar se o polimorfismo Ala16Val da enzima antioxidante superóxido dismutase dependente de manganês (SOD2), que afeta a eficiência detoxificadora da enzima, poderia ter efeito sobre a resposta citotóxica ao MTX. Para tanto, foi realizado um estudo in vitro utilizando células mononucleares do sangue periférico (CMSP), obtidas de doadores saudáveis portadores de diferentes genótipos do polimorfismo Ala16Val-SOD (genótipos = AA, VV e AV). Uma vez obtidas, as CMSPs foram tratadas com MTX nas concentrações de 10 e 100 μM por 24 e 72 horas, sendo posteriormente analisado o efeito sobre a viabilidade, modulação do metabolismo oxidativo-inflamatório e apoptótico. As CMSP-AA, que naturalmente apresentam uma SOD2 30 a 40% mais eficiente do que as CMSP-VV, apresentaram maior resistência ao tratamento com MTX em relação às CMSP-AV/VV. Quanto aos níveis de produção de EROS (espécies reativas de oxigênio) e lipoperoxidação, houve aumento significativo nas células expostas ao MTX independente do genótipo, entretanto, o aumento dos níveis de carbonilação de proteínas foi observado apenas em CMSP-AV/VV. Nas CMSP-AA houve diminuição da atividade da SOD2. Já quanto aos níveis de Glutationa Peroxidase, as CMSP-AA apresentaram uma elevação mais intensa. Os níveis das caspases 3 e 8 foram aumentados nas CMSP expostas ao MTX, mas a modulação desses genes, assim como do Bax e Bcl-2 (genes envolvidos rota apoptótica), foi genótipo-dependente. O MTX foi capaz de elevar os níveis das citocinas inflamatórias e diminuir o nível da citocina antiinflamatória IL-10, independente do genótipo. Os resultados sugerem que o polimorfismo Ala16Val-SOD2 é capaz de modular a resposta citotóxica de CMSP ao MTX.

O consumo de álcool tem sido considerado um problema de saúde pública. Entretanto, também existem evidências de eventuais benefícios do consumo leve a moderado de algumas bebidas alcoólicas, no que se refere a doenças crônicas não transmissíveis. As doenças periodontais, sendo a sexta doença crônica mais prevalente, também têm sido associadas ao consumo de álcool com resultados controversos. Esta tese avaliou, através de três artigos a relação entre consumo de álcool e doenças periodontais. O primeiro artigo de divulgação, tem por objetivo alertar a profissão a respeito do estado atual do conhecimento em relação a associação entre álcool e doenças periodontais. O segundo artigo é um estudo experimental em modelo animal que avaliou o efeito da dependência química álcool sobre desfechos periodontais. Os resultados não mostraram diferenças estatisticamente significativas na perda óssea alveolar e secreção de TNF-α. O terceiro artigo, também em modelo animal, abordou o consumo do vinho tinto e valeu-se de controles total, álcool a 12%, suco de uva e resveratrol. A análise de ocorrência de periodontite espontânea demonstrou que o vinho tinto tem potencial protetor para a perda óssea alveolar e secreção de TNF- α . Os resultados encontrados nessa tese são instigantes e estão em linha com a literatura que demonstra uma relação do tipo curva J entre álcool e doenças periodontais, na qual até um determinado nível, o fator comporta-se como protetor. Assim, essa relação deve ser considerada na abordagem clínica, sem incentivo ao consumo, mas compreendendo até onde este é tolerável e/ou benéfico no que se refere a doenças periodontais. / Alcohol consumption has been considered a public health problem. Otherwise, there is evidence of eventual benefits of a light to moderate consume of some alcoholic beverage, in relation to non-transmissible chronic disease. The periodontal diseases, being the sixth most prevalent chronic disease, also have been associated to alcohol consumption with controversial results. This thesis evaluated, through three articles the relationship between alcohol consumption and periodontal diseases. The first article has the objective of alerting the professionals about the actual knowledge about the relationship between alcohol and periodontal diseases. The second article is an experimental study in animal model which evaluated the alcohol dependence over periodontal outcomes. The results did not show statistically significant differences in alveolar bone loss and TNF- α secretion. The third article, also in animal model, studied the red wine consumption and used total controls, 12% alcohol, grape juice and resveratrol. The analysis of the occurrence of spontaneous periodontitis demonstrated that red wine has a protector potential for alveolar bone loss and TNF- α secretion. The results found in this thesis are curious and they are in line with the literature that demonstrates a relationship like J-curve between alcohol and periodontal diseases, until some level, the factor behave as protector. So, this relation must be considered in clinical approaches, without encouraging the consumption, but understanding how much is tolerable and/or benefit according to periodontal diseases.

In Brazil, the last national population survey, observed that the prevalence of obesity was 20.3% among individuals over 18 years of age, when analyzed according to sex and schooling, it is observed that women with lower levels of schooling have a higher prevalence 25, 2%. Height in adults is a widely used nutritional marker, which reflects the interaction between genetic inheritance and environmental exposure throughout life, so for an assessment of the broad nutritional status it is also necessary to assess height. The prevalence of obesity-related comorbidities is more pronounced in populations with lower height, with an increase in blood pressure, LDL-cholesterol levels, atherosclerotic plaque numbers, prevalence of diabetes mellitus and reduction of HDL- Cholesterol in adults. Obesity is characterized by a low-grade chronic inflammatory picture, leading to inflammatory responses related to excess body fat resulting from increased local and systemic production of cytokines and adipokines, including C-reactive protein (CRP), tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), leptin and plasminogen activator inhibitor 1, and reduction of adiponectin and interleukin 10 (IL-10). These act as a trigger in the progression of atherosclerosis. Aiming to contribute to the discussion of the problem, this dissertation presents: a review chapter on obesity and the mechanisms of physiological adaptation due to malnutrition in the beginning of life; The second article refers to a non-random clinical trial with a three-month follow-up that compared the metabolic and inflammatory profile of obese women with short stature and without short stature submitted to interdisciplinary therapy for weight loss. / FAPEAL - Fundação de Amparo à Pesquisa do Estado de Alagoas / No Brasil, o último levantamento populacional nacional, observou que a prevalência de obesidade foi de 20,3% entre indivíduos acima de 18 anos de idade, quando analisada segundo sexo e escolaridade observa-se que as mulheres com menor escolaridade apresentam maior prevalência 25,2%. A altura em adultos é um marcador nutricional amplamente utilizado, que reflete a interação entre herança genética e a exposição ambiental ao longo da vida, assim para uma avaliação do estado nutricional ampla é necessário também avaliar altura. Observa-se que nas populações com menor altura a prevalência das comorbidades relacionadas a obesidade são mais acentuadas, observando aumento da pressão arterial, dos níveis de LDL-colesterol, do número de placa aterosclerótica, prevalência de diabetes mellitus e redução dos níveis de HDL-colesterol em adultos. A obesidade é caracterizada por quadro inflamatório crônico de baixo grau, levando as respostas inflamatórias relacionadas ao excesso de gordura corporal resultante do aumento da produção local e sistêmica de citocinas e adipocinas, incluindo a proteína C reativa (CRP), o fator de necrose tumoral α (TNF-α), a interleucina 6 (IL-6), a interleucina 8 (IL-8), a leptina e o inibidor do ativador do plasminogênio 1 e redução da adiponectina e na interleucina 10 (IL-10). Esses atuam como um gatilho na progressão da aterosclerose. Visando contribuir com a discussão do problema, esta dissertação apresenta: um capítulo de revisão sobre obesidade e os mecanismos de adaptação fisiológicos em decorrência da má nutrição no início de vida; o segundo artigo refere-se a um estudo de ensaio clinico, não aleatório com três meses de seguimento que comparou o perfil metabólico e inflamatório de mulheres obesas com baixa estatura e sem baixa estatura submetidas à terapia interdisciplinar para perda de peso.

Orientadores: Francisco Eduardo Martinez, Wellerson Rodrigo Scarano / Texto em português e inglês / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T05:09:17Z (GMT). No. of bitstreams: 1 Mendes_LeonardodeOliveira_D.pdf: 2740474 bytes, checksum: 7720a25e258ca28197be2547f35bc162 (MD5) Previous issue date: 2014 / Resumo: O etanol provoca danos reprodutivos, diretamente nos tecidos ou indiretamente via eixo hipotalâmico-hiposário-gonadal. A próstata, hormônio-dependente, é susceptível aos efeitos do etanol. Evidências atribuem papel anti-inflamatório à testosterona, com crescente interesse na ação da terapia hormonal sobre o etanol. Assim, o presente trabalho teve como objetivo avaliar os efeitos da terapia hormonal com testosterona sobre o consumo de etanol: 1) nas dosagens hormonais plasmátcas e tecidual, na histopatologia, proliferação celular e na localização e expressão de receptores hormonais (artigo I); 2) nas concentrações plasmáticas e expressão de citocinas pró e anti-inflamatórias (artigo II) no lobo ventral da próstata de ratos UChB. Ratos com 90 dias de idade foram divididos em dois grupos experimentais (n = 20/grupo): C, consumo de água ad libitum,e EtOH, consumo de etanol a 10% (v/v), > 2g de etanol/kg de peso corpóreo/dia, e água ad libitum. Aos 150 dias de idade, 10 ratos de cada grupo receberam injeções subcutâneas de cipionato de testosterona (5mg/kg de peso corpóreo) diluídos em olho de milho durante quatro semanas em dias alternados, T e EtOH+T, enquanto os restantes (10/grupo) receberam somente óleo de milho como veículo. Aos 180 dias de idade, os ratos foram eutanasiados por decapitação para coleta do sangue e as próstatas ventrais foram dissecadas, pesadas e processadas. Secções da próstata ventral foram coradas com hematoxilina e eosina, impregnação pela prata e azul de toluidina. Radioimunoensaio para avaliação das concentrações plasmáticas de testosterona, estradiol, diidrotestosterona (DHT) e testosterona intraprostática, imuno-histoquímica para Ki-67, AR, ER?, ER?, DACH-1,TGF-?1, pSmad 2, e-caderina e ?-actina, western blot para AR, ER?, ER?, DACH-1, PAR4, IL-6 IL-10, TNF? e TGF-?1 e elisa para determinação das concentrações plasmáticas de IL-6, IL-10, TNF? e TGF-?1foram realizados. A terapia com testosterona aumentou o peso da próstata ventral. Houve diminuição do compartimento epitelial e aumento do luminal no EtOH com a terapia hormonal revertendo esses efeitos. O compartimento estromal do EtOH caracterizou-se pela presença de feixes de fibras reticulares delgados e esparsos, com ruptura da camada de células musculares lisas.Focos inflamatórios, metaplasia, atipia reativa inflamatória, perda de adesão entre as células epiteliais e aumento de mastócitos intactos e desgranulados foram frequentes no EtOH e ausentes no EtOH+T. Tanto testosterona como etanol não alteraram a taxa de proliferação celular, porém maior expressão de PAR4 foi observada no EtOH. Houve aumento da testosterona plasmática, intraprostática e DHT no T e EtOH+T e diminuição do estradiol plasmático no EtOH+T. Os T e EtOH+T também apresentaram níveis plasmáticos superiores de TNF? e TGF-?1, sem diferença em relação à IL-6 e IL-10. A expressão de AR, ER?, DACH-1 e IL-6 na próstata foi semelhante entre os grupos experimentais, diferentemente do que foi observado para ER?, TNF? e TGF-?1, que diminuíram após terapia hormonal.Nossos resultados indicam que o etanol foi capaz de induzir a emergência de focos inflamatórios, além de suprimir a imunorreatividade para e-caderina e ?-actina. A testosterona reverteu esses efeitos, reduzindo a expressão do ER?, TGF-?1, TNF? e NFR2, tornando-se um possível alvo a ser avaliado para as disordens causadas pelo etanol / Abstract: Ethanol induces reproductive damages, directly in the tissues or indirectly by hormonal imbalance. Prostate, a hormone-dependent gland, is susceptible to effects caused by ethanol. Emerging evidences assign to testosterone an anti-inflammatory role, with growing interest in the action of hormone therapy on the effects of ethanol. Therefore, the current research aimed to assess the effects of hormone therapy with testosterone on the ethanol consumption: 1) in tissue and plasma hormone assays, histopathology and immunolocalization and expression of hormone receptors (manuscript I); plasma levels and expression of pro and anti-inflammatory cytokines (manuscript II) in the ventral prostate of UChB rats (ethanol-preferring rat). UChB rats aged 90 days were divided into two experimental groups (n=20): C: drinking water only and EtOH: drinking 10% (v/v) ethanol at > 2 g/kg body weight/day + water. At 150 days of age, 10 rats from each group received subcutaneous injections of testosterone cypionate (5mg/kg body weight) diluted in corn oil every other day during 4 weeks, constituting T and EtOH+T, while the remaining animals (10/group) received corn oil as vehicle. All animals were euthanized at 180 days old by decapitation. Blood was collected to obtain plasma hormone and cytokines concentrations and ventral prostate was dissected, weighted and processed. Prostate sections were stained with hematoxylin and eosin, Gomori¿s reticulin and toluidine blue. The following techniques were performed: radioimmunoassay to plasma concentrations of testosterone, dihydrotestosterone (DHT), estradiol and intraprostatic testosterone, immunohistochemistry to Ki-67, AR, ER?, ER?, DACH-1, TGF-?1, pSmad 2, e-cadherin e ?-actinin, western blot to AR, ER?, ER?, DACH-1, PAR4, IL-6 IL-10, TNF? and TGF-?1 and elisa to plasma concentrations of IL-6, IL-10, TNF-? and TGF-?1. Testosterone therapy increased the ventral prostate weight. There were reducing of epithelial compartment and increasing of luminal compartment in the EtOH and hormonal therapy was able to reverse this pattern. Stroma compartment of EtOH showed thin and sparse reticular fiber bundles with rupture of smooth muscle cell layer. Inflammatory foci metaplasia, inflammatory reactive atypia, loss of cell-cell adhesion and increasing of degranulated mast cells were frequent in EtOH and absent in EtOH+T. Testosterone and ethanol did not alter the proliferation rate, but high expression of PAR4 was shown in EtOH. There was increasing of plasma and intraprostatic testosterone and plasma DHT in T and EtOH+T and decreasing of estradiol in EtOH+T. The T and EtOH+T exhibited increasing of plasma TNF? and TGF-?1, without differences regarding to IL-6 and IL-10. AR, ER?, DACH-1 and IL-6 expressions were similar among the experimental groups, differently from ER?, TNF? and TGF-?1, which decreased after hormonal therapy. Our results show that ethanol was able to induce the emergence of inflammatory foci, besides suppress the immunorreactivity to e-cadherin and ?-actinin. Testosterone was able to reverse these effects, downregulating ER?, TGF-?1, TNF? and NFR2, showing to be promising in the treatment of alcohol-related disorders / Doutorado / Biologia Tecidual / Doutor em Biologia Celular e Estrutural

Much previous work on air pollution epidemiology has studied end-stage outcomes such as mortality or severe ill health warranting emergency admission, often based on clinical criteria prone to misclassification, and usually without accompanying study of the mediating mechanisms. Therefore this work has three specific objectives: firstly, to assess the effects of short-term exposure to particles and gases on acute coronary syndrome (ACS), by measuring the levels of cardiac troponin T (cTnT), a highly sensitive and specific marker of myocardial damage in patients admitted to hospital for chest pain of myocardial origin; secondly, to investigate the effects of short-term changes in ambient air pollution on the occurrence of transient ischaemic attacks (TIA); finally, to investigate the effects of ambient and personal exposure to air pollutants on a range of mediators or markers in a putative susceptible population. Two case-crossover studies were conducted to study the association between short-term changes in air pollutants and ischaemic cardiac events and TIA. Hospital data on admissions were analysed for actual or suspected ischaemic events and the associated cTnT levels were obtained. For the TIA project, data on 709 subjects were obtained from five TIA centres clustered around Manchester and Liverpool. In the third project a panel of 35 type 2 diabetes mellitus patients were prospectively followed fortnightly for a total of four visits. At each visit blood was collected to measure markers of inflammation, coagulation and endothelial function. In all three projects ambient air pollution data were obtained from background monitoring networks and in the third project personal exposure to PM2.5 was measured. Project 1: Of 28,622 admissions, 17.5% were ACS with myocyte necrosis (cTnT 0.03-1ng/ml) and 1004 (3.5%) were cases of myocardial infarction (cTnT ≥ 1 ng/ml). Both particulate and gaseous pollutants were associated with admission for ACS. The two largest effects per interquartile increase of exposure were observed with PM10 with ORof 1.14 (95% CI: 1.05-1.24) and with SO2, OR 1.11 (95% CI: 1.00-1.23). Associations between pollution and ACS admissions were the strongest for women, those above the age of 65 years and in the cooler season. Project 2: In the Manchester dataset, exposure to nitric oxide (NO) was associated with occurrence of TIA, while no effect was observed for Liverpool data. Subgroup analysis reveals that CO, NO and NO2 were more strongly related to the occurrence of TIA in participants above the age of 65 years and male patients. Project 3: No consistent association was observed between measured biomarkers and air pollutants using exposure data from ambient monitoring stations. In contrast, significant association between personal PM2.5 and interleukin-6 (IL-6) was observed. Similarly, personal PM2.5 had large but non-significant positive associations with high sensitivity C-reactive protein and fibrinogen. The results of this study reveal that short-term changes in particulate and gaseous pollution are related to the risk of admission for ACS as demonstrated by a specific marker hitherto not used for this purpose. It provides limited evidence for an association between changes in ambient NO concentration (which may have been a surrogate for another pollutant), and the occurrence of TIA, which had not previously been studied as an air pollution outcome, and increase in IL-6, a major pro-inflammatory marker. The IL-6 response to personal PM2.5 provides evidence in support of the link between ambient levels of particles/gases and cardiopulmonary morbidity and mortality.

Epidemiological studies indicate long-term risks of ionizing radiation on the heart, even at moderate doses. In this study, we investigated the inflammatory, thrombotic and fibrotic late responses of the heart after low-dose irradiation (IR) with specific emphasize on the dose rate. Hypercholesterolemic ApoE-deficient mice were sacrificed 3 and 6 months after total body irradiation (TBI) with 0.025, 0.05, 0.1, 0.5 or 2 Gy at low (1 mGy/min) or high dose rate (150 mGy/min). The expression of inflammatory and thrombotic markers was quantified in frozen heart sections (CD31, E-selectin, thrombomodulin, ICAM-1, VCAM-1, collagen IV, Thy-1, and CD45) and in plasma samples (IL6, KC, MCP-1, TNFα, INFγ, IL-1β, TGFβ, INFγ, IL-10, sICAM-1, sE-selectin, sVCAM-1 and fibrinogen) by fluorescence analysis and ELISA. We found that even very low irradiation doses induced adaptive late responses, such as increases of capillary density and changes in collagen IV and Thy-1 levels indicating compensatory regulation. Slight decreases of ICAM-1 levels and reduction of Thy 1 expression at 0.025–0.5 Gy indicate anti-inflammatory effects, whereas at the highest dose (2 Gy) increased VCAM-1 levels on the endocardium may represent a switch to a pro-inflammatory response. Plasma samples partially confirmed this pattern, showing a decrease of proinflammatory markers (sVCAM, sICAM) at 0.025–2.0 Gy. In contrast, an enhancement of MCP-1, TNFα and fibrinogen at 0.05–2.0 Gy indicated a proinflammatory and prothrombotic systemic response. Multivariate analysis also revealed significant age-dependent increases (KC, MCP-1, fibrinogen) and decreases (sICAM, sVCAM, sE-selectin) of plasma markers. This paper represents local and systemic effects of low-dose irradiation, including also age- and dose rate-dependent responses in the ApoE-/- mouse model. These insights in the multiple inflammatory/thrombotic effects caused by low-dose irradiation might facilitate an individual evaluation and intervention of radiation related, long-term side effects but also give important implications for low dose anti-inflammatory radiotherapy.