Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-17T21:14:35Z No. of bitstreams: 1 AnaGiseliaNascimento.pdf: 3146962 bytes, checksum: 2e04b37893415a3b55c8a3bcd2cf19b9 (MD5) / Made available in DSpace on 2017-05-17T21:14:35Z (GMT). No. of bitstreams: 1 AnaGiseliaNascimento.pdf: 3146962 bytes, checksum: 2e04b37893415a3b55c8a3bcd2cf19b9 (MD5) Previous issue date: 2016-09-30 / Pituitary adenomas represent 10-15% of all intracranial tumors with an incidence of 1/1000 of the population. They are considered benign and mostly slow growth. Despite this, about 35% may present with invasion of parasellar compartments and a significant number have a clinically aggressive behavior with early recurrence during follow-up, despite surgery and other therapeutic approaches. Such aggressive tumors should be identified early in order to provide a more incisive treatment and / or a strict clinical and radiological follow-up, with prompt djunctive therapy institution in persistence or recurrence of these tumors. Despite these considerations, to date, no clinical, radiological and histological marker predicting such behavior has been established. This study aims to reassess the role of Ki-67 through an improved immunohistochemical technique for the expression of these antigens and correlate it to demographic and clinicopathological parameters of pituitary adenomas. This is a crosssectional study conducted from November 2015 to August 2016 were used pituitary adenoma tissue from 62 patients from the Endocrinology Service of the University of the Federal University of Maranhão Hospital. For evaluation of adenomas of medical records data were analyzed, namely: clinical and serum levels of related hormones as well as magnetic resonance imaging of the pituitary gland. To analyze the expression of Ki-67 was performed immunohistochemical technique using the anti-human monoclonal primary antibody, expressed by the percentage of labeled nuclei in the form of labeling index Ki-67. In our sample of 62 patients, 31 patients were female (50%) with a mean age at surgery of 46.2 (± 13.3) years, ranging from 16 to 73 years. Ki-67 index, as well as their median had to be higher in females, with p = 0.04 and p = 0.02, respectively. There was no predominance of expression and Ki-67 index with respect to age and diagnosis. Adenomas giant had higher median Ki-67 with respect to non giants with p = 0.04, but there was no correlation of Ki-67 with tumor invasion. The pituitary adenomas previously submitted to specific pharmacotherapy showed expression of Ki-67 lower, however, only when the somatotropinomas were analyzed separately, there was statistically significant with p = .0,03. The Ki-67 antigen, a marker of tumor proliferative activity, play an important role in pituitary adenomas. In conjunction with other biomarkers such as tumor invasion, large volume adenoma the initial presentation of patients and / or rapid tumor growth rate documentation can lead to identification of patients which show an aggressive clinical behavior, in order to conduct a multimodal therapeutic approach more effective or more strict postoperative follow-up. / Adenomas hipofisários representam de 10-15% de todas as neoplasias intracranianas com prevalência de um caso para 1000 habitantes. Apresentam grande variação de comportamento clínico, sendo a maioria de crescimento indolente, enquanto cerca de 35% podem ser invasivos e um pequeno grupo francamente agressivo. Até o presente momento, não há um biomarcador que possa predizer seu comportamento de forma confiável, o que possibilitaria uma terapia adjuvante mais agressiva ou um seguimento clínico mais rigoroso. O objetivo desse estudo foi avaliar a expressão e o índice do marcador imuno-histoquímico de proliferação celular, antígeno Ki-67 em adenomas hipofisários e correlacioná-lo a parâmetros demográficos e clínico-patológicos, visando definir sua relevância prognóstica. Trata-se de um estudo transversal realizado no período de novembro de 2015 a agosto de 2016. Foram utilizados tecidos de adenomas hipofisários de 62 pacientes oriundos do Serviço de Endocrinologia do Hospital Universitário da Universidade Federal do Maranhão. Para avaliação dos adenomas foram analisados dados dos prontuários, a saber: avaliação clínica e dosagens séricas de hormônios relacionados, assim como ressonância nuclear magnética de hipófise. Para analisar a expressão do Ki-67, foi realizada a técnica de imuno-histoquimica utilizando-se o anticorpo primário monoclonal anti-humano expressado pela percentagem de núcleos imunopositivos sob forma de índice de marcação de Ki-67. Na amostra de 62 pacientes, 31 pacientes foram do sexo feminino (50%) com a média de idade à época da cirurgia de 46,2 (±13,3) anos, variando de 16 a 73 anos. O antígeno Ki-67 apresentou expressão em 37 (59,7%) pacientes, variando o seu índice de 0,1% a 2,4%. O índice Ki-67, assim como a sua mediana apresentaram-se mais elevados no sexo feminino com p=0,04 e p=0,02, respectivamente. Não houve predominância da expressão e do índice de Ki-67 com relação a idade e diagnóstico. Adenomas gigantes apresentaram mediana de Ki-67 mais elevada com relação aos não gigantes com valor de p=0,04, mas não houve qualquer correlação de Ki-67 com a invasão tumoral. Os adenomas hipofisários previamente submetidos à farmacoterapia específica apresentaram expressão de Ki-67 mais baixos, no entanto, somente quando os somatotropinomas foram analisados de forma isolada, houve significância estatística com p=0,03. O antígeno Ki-67, marcador de atividade proliferativa tumoral, tem papel relevante em adenomas hipofisários. Em conjunto com outros biomarcadores, como invasão tumoral, grande volume do adenoma à apresentação inicial dos pacientes e/ou documentação de rápida velocidade de crescimento do tumor, pode levar a identificação de pacientes que demonstrem um comportamento clínico agressivo, de forma a conduzir a uma abordagem terapêutica multimodal mais efetiva ou a um seguimento pós operatório mais estrito.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-17T21:24:47Z No. of bitstreams: 1 NadjaCruzGaido.pdf: 2766927 bytes, checksum: 6fdfd1a4a2beb11f1da8d3669b11750a (MD5) / Made available in DSpace on 2017-05-17T21:24:47Z (GMT). No. of bitstreams: 1 NadjaCruzGaido.pdf: 2766927 bytes, checksum: 6fdfd1a4a2beb11f1da8d3669b11750a (MD5) Previous issue date: 2016-09-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) / The pituitary adenomas represent about 10 to 15% of intracranial tumors and are usually benign, comprising a cell population of monoclonal origin. They are classified as macroscopic and radiological aspects, functional status, dyeing, immunohistochemistry and microscopic. About 70% of human cancers have mainly a deficiency in the function gene. The p53 gene is extensively studied in tumors, showing that patients with mutations have a worse prognosis, as well, the p53 protein is critical in preventing tumor development performing change detection function in DNA and thereby fix or apoptosis. It is possible that the increase in expression of this protein is the result of an attempt to stop the cell cycle in response to deregulation by a stimulus from another source. This way, the study seeks to determine the expression of p53 protein in pituitary adenomas. an analytical study, with crosssectional was held in which were included 62 patients older than 16 years of both sexes, who were diagnosed with pituitary adenoma, from 2008 to 2016, arising from the Endocrinology Service University hospital of the Federal University of Maranhão - HUUFMA. These patients underwent surgical resection of the tumor, obtaining the samples of tumor tissue. After conventional histopathology, they were embedded in paraffin, for carrying out the immuno-histoqumico study aimed to identify the expression of p53 protein and implications correlated with biological behavior of pituitary adenomas. The analysis for p53 protein expression in adenomas was obtained by immunoenzymatic of streptoavidin-biotin or immunohistochemistry. The p53 protein immunostaining revealed no statistically significant correlation, as the clinical and demographic characteristics such as gender, age, p53 expression in tumor subtypes and tumor volume. It is necessary to obtain greater numbers of patients, so that p53 expression is effective when the real prognostic value in pituitary adenomas. / Os adenomas hipófisários representam cerca de 10 a 15% das neoplasias intracranianas e são geralmente benignos e compostos por uma população celular de origem monoclonal. São classificados conforme aspectos macroscópicos e radiológicos, status funcional, tintoriais, imuno-histoquímica e microscópicas. Cerca de 70% dos cânceres humanos possuem principalmente uma deficiência na função do gene. O gene p53 é extensivamente estudado nas neoplasias mostrando que pacientes com mutações apresentam um pior prognóstico, assim, a proteína p53 é fundamental na prevenção do desenvolvimento de tumores exercendo a função de detecção de alterações no DNA e, consequentemente, correção ou apoptose. É possível que o aumento na expressão desta proteína seja decorrente de uma tentativa de frear o ciclo celular como resposta à desregulação por um estímulo de outra origem. Dessa forma, o estudo busca avaliar a imunoexpressão da proteína p53 em adenomas hipofisários. Foi realizado um estudo analítico com delineamento do tipo transversal, no qual foram incluídos 62 pacientes com idade superior 16 anos de ambos os sexos, que apresentaram diagnóstico de adenoma hipofisário no período de 2008 a 2016, oriundos do Serviço de Endocrinologia do Hospital Universitário da Universidade Federal do Maranhão – HUUFMA. Esses pacientes foram submetidos à ressecção cirúrgica do tumor, com obtenção das amostras do tecido tumoral. Após diagnóstico histopatológico convencional, foram emblocadas em parafina, para realização do estudo imuno-histoqumico com o objetivo de identificar a imunoexpressão para proteína p53 e implicações correlacionadas ao comportamento biológico dos adenomas hipofisários. A análise para expressão da proteína p53 em adenomas hipofisários foi obtida através da reação imunoenzimática da streptoavidina-biotina-peroxidase ou imuno-histoquimica. A imunoexpressão da proteína p53 não revelou correlação estatisticamente significante, quanto aos achados clínicos e características demográficas, tais como sexo, idade, imunoexpressão da p53 para os subtipos tumorais e volume tumoral. Torna-se necessário obter números mais expressivos de pacientes para que a imunoexpressão da p53 seja efetiva, quanto ao real valor prognóstico em adenomas hipofisários.

A partir da hipófise de Astyanax altiparanae foram clonados e caracterizados os cDNAs completos para fshb, lhb e gpha. Das gônadas, foram clonados os cDNAs para fshr e lhr. As análises filogenéticas destes genes demonstram uma proximidade com as sequências homólogas das ordens Siluriformes e Cypriniformes. Por meio de qPCR, detectamos que a expressão dos mRNAs para as gonadotropinas foi restrita à hipófise, ao passo que dos receptores foi restrita às gônadas. Foi detectado que os mRNAs/proteínas para subunidades beta são expressas por diferentes populações de células gonadotrópicas. Nas gônadas, ambos receptores foram localizados nas células de Sertoli e Leydig, bem como nas células foliculares e da teca de oócitos a partir de crescimento secundário. Então, os níveis de expressão gênica das subunidades das gonadotropinas e seus receptores, juntamente como os níveis plasmáticos de esteroides sexuais foram avaliados e relacionados com a cinética da gametogênese durante o ciclo reprodutivo anual de machos e fêmeas em cativeiro. / From Astyanax altiparanae pituitaries, we cloned the full-leght cDNAs coding for fshb, lhb e gpha. From gonads, we cloned the full-length cDNAs for fshr and lhr. Phylogenetic analysis revealed that all five sequences showed the highest identity degree with homologous sequences from Siluriformes and Cypriniformes, respectively. Using qPCR, we observed that gonadotropin subunits expression were restricted to the pituitary gland, just like their receptors were restricted to the gonads. We also observed that both gonadotropin beta subunits mRNAs/proteins are expressed by distinct populations of gonadotropes at pituitary. In the gonads, both receptors were detected on Sertoli and Leydig cell, as well as on ovaries were located on follicular and theca cells, in oocytes starting from secondary growth. Then, gene expression levels of gonadotropins subunits mRNAs and their receptors were assessed, in parallel to plasma level of sex steroids, and evaluated according to gemetogenesis during the annual reproductive cycle of males and females kept in captivity.

Les écosystèmes marins constituent une cible majeure des changements globaux qui affectent de façon pérenne notre planète.Nous nous sommes intéressés à deux de ces changements susceptibles de menacer le milieu marin : le réchauffement climatique par le biais de l’étude de l’effet du stress thermique sur l’expression de ho-1 et la pollution via l’étude de l’impact des métaux lourds à effets perturbateurs endocriniens (le cadmium et le plomb) sur l’axe hypothalamo-hypophyso-gonado-hépatique (HHGH) chez le loup (Dicentrarchus labrax L.).Nos résultats montrent une réponse importante de ho-1 aux stress thermique et chimique dans le foie. Nous avons également démontré une forte accumulation hépatique du Cd, et à un degré moindre du Pb, accompagnée d’une sur-expression du gène mt codant pour les métallothionéines. Des variations dans l’expression de gènes clés le long de l’axe HHGH (arom b, fshß, arom a…) ont été observées après intoxication par les deux métaux sans pour autant induire des effets physiologiques observables.La question se pose de savoir si la synergie des stress physico-chimiques impacte la dynamique et l’état sanitaire des populations marines. / Marine ecosystems are a major target of global changes that continuously affect our planet. In the present study we investigated two of these changes that may threaten the marine environment: global warming, through the study of the effect of heat stress on ho-1 expression and marine pollution through the study of the impact of heavy metals acting as endocrine disruptors (cadmium and lead) on the hypothalamic-pituitary-gonadal-liver (HHGL) axis in the sea bass (Dicentrarchus labrax L.). Our results show an important response of ho-1 to both thermal and chemical stress in the liver. We also demonstrated a high hepatic accumulation of Cd, and to a lesser extent of Pb. This accumulation was correlated with an overexpression of mt gene coding for metallothionein. Changes in the expression of candidate genes (arom b, fshß, arom a…) along the HHGL axis were observed after metal intoxication but no physiological effects were observed.The question then arises to what extent the synergy of physicochemical stressors impacts the dynamics and the welfare of marine species.

The first goal of the current research was to determine whether gene expression profiles differed between whole pituitaries of growing beef steers grazing pastures containing high (HE) or low (LE) amounts of toxic endophyte-infected tall fescue. The global (microarray analysis) and selected targeted (RT-PCR) mRNA expression patterns of pituitaries collected from beef steers (BW = 266 ± 15.5 kg) that had been randomly assigned to undergo summer-long grazing (89 to 105 d) of either HE (0.52 ppm ergot alkaloids) or LE (< 0.03 ppm ergot alkaloids) pastures were compared. Gene expression data were subjected to one-way ANOVA. The pituitaries of HE steers had 542 differentially expressed genes, and the pattern of altered gene expression was dependent on treatment. Targeted RT-PCR analysis corroborated these findings, including decreased expression of DRD2, PRL, POU1F1, GAL, and VIP and that of POMC and PCSK1, respectively. Canonical pathway analysis (Integrated Pathway Analysis, IPA) identified HE-dependent alteration in signaling of additional pituitary-derived hormones, including growth hormone and GnRH. In conclusion, consumption of endophyte-infected tall fescue alters the pituitary transcriptome profiles of steers in a manner consistent with their negatively affected physiological parameters. The second goal of this project was to test the hypothesis that sodium selenite (ISe), SEL-PLEX (OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter pituitary transcriptome profiles in growing beef steers (BW = 183 ± 34 kg) commonly grazing an endophyte-infected tall fescue (HE) pasture. Steers were randomly selected from herds of fall-calving cows grazing HE pasture and consuming VM mixes that contained 35 ppm Se as either ISe, OSe, or MIX forms. Steers were weaned, depleted of Se for 98 d, and subjected to summer-long common grazing of a 10.1 ha HE pasture containing 0.51 ppm ergot alkaloids. Steers were assigned (n = 8) to the same Se-form treatments on which they were raised. Selenium treatments were administered by daily top-dressing 85 g of VM mix onto 0.23 kg soyhulls, using in-pasture Calan gates. Pituitaries were collected at slaughter and changes in global (microarray) and selected (RT-PCR) mRNA expression patterns determined. The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The form of Se affected the expression of 542 annotated genes. Integrated Pathway Analysis found a canonical pathway network between prolactin and POMC/ACTH/ α-MSH synthesis-related proteins, and that mitochondrial dysfunction was a top-affected canonical pathway. Targeted RT-PCR analysis found that the relative abundance of mRNA encoding prolactin and POMC/ACTH/ α-MSH synthesis-related proteins was affected by the form of Se, as were mitochondrial dysfunction-related proteins OSe steers appeared to have a greater prolactin synthesis capacity vs. ISe steers through decreased dopamine receptor D2 signaling, whereas MIX steers had a greater prolactin synthesis capacity and release potential by increasing TRH concentrations than ISe steers. OSe steers also had a greater ACTH and α-MSH synthesis potential than ISe steers. We conclude that form of Se in VM mixes affected genes responsible for prolactin and POMC/ACTH/α-MSH synthesis, and mitochondrial function in pituitaries of growing beef steers commonly grazing an HE pasture. The third goal was to test the hypothesis that sodium selenite (ISe), SEL-PLEX (OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter selenoprotein profiles in pituitaries and livers of growing beef steers commonly grazing an endophyte-infected tall fescue (HE) pasture (i.e., the same steers used in Goal 2). The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The mRNA content of 6 selenoproteins in the pituitary was affected by Se treatments, along with two selenoprotein P receptors, whereas the expression of two selenoproteins was altered in the liver. We conclude that the change in selenoprotein gene expression in pituitaries indicates that OSe steers have a greater potential capacity to manage against oxidative damage, maintain cellular redox balance, and have a better quality control of protein-folding in their pituitaries than ISe steers. The change in selenoprotein gene expression by the liver indicates that MIX steers have a greater redox signaling capacity and capacity to manage oxidative damage than ISe steers.

Abstract not available

Abstract not available

Includes bibliographical references (leaves 228-257). Aims to determine the impact of maternal undernutrition during late gestation and during the periconceptional and gestational periods on fetal growth, fetal blood pressure and the fetal hypothalamo-pituitary adrenal axis in the sheep.

Low birth weight is associated with an increased risk of impaired glucose tolerance and type 2 diabetes and with signs of increased hypothalamic pituitary adrenal axis activity in later life (1, 2). Low birth usually weight reflects a reduction in fetal growth, which largely depends on an adequate supply of nutrients and oxygen. Variations in supply modify the metabolic and neuroendocrine characteristics of the fetus, which in turn modulate the pattern of functional development as well as growth (3). An adverse fetal environment, evident as low birth weight, is therefore proposed to alter functional development with long term effects for the function and risk of disease in the individual later in life (4, 5). Increased HPAA impairs metabolic homeostasis and could therefore mediate effect of prenatal challenge on later metabolic control (6). It was therefore hypothesised that restriction of fetal growth, increases circulating cortisol and/or alters sensitivity to cortisol, which increases fasting blood glucose, and impairs glucose tolerance in the young adult. Large litter size in the guinea pig is characterised by reduced placental and fetal growth, reduced size at birth and insulin resistance in offspring in later life, providing a suitable model to test this hypothesis. Spontaneous restriction of fetal growth in the guinea pig, evident as small size at birth, was associated with increased salivary cortisol, in both sexes but at different stages of postnatal life. In males, salivary cortisol was increased with small size at birth in early and adult life, but reduced later with ageing. In females however, salivary cortisol was increased in juveniles and in aged adults, possibly reflecting the impact of the oestrus cycle on cortisol production in mature cycling females. Altered activity of the HPGA, which can influence that of the HPAA, has also been reported to be programmed by prenatal restriction. In the guinea pig, salivary testosterone in males increased with age and small size at birth in juveniles, young and aged adults. In females, salivary progesterone increased with age up to 300 days, and decreased with size at birth in the young guinea pig. Although testosterone inhibits HPAA activity, in males, mean salivary cortisol correlated positively with mean salivary testosterone at 100 and 300 days of age. In contrast, progesterone may enhance HPAA activity, and consistent with this, in females, mean salivary progesterone correlated with mean salivary cortisol at 400 days of age. Therefore, salivary testosterone in the male and salivary progesterone in the female guinea pig changes with maturation and has previously reported in this or other species, but small size at birth increases salivary testosterone in males with modest effects in early life in females. This together with the unexpected positive associations of salivary cortisol with testosterone in males, suggests that programming of the HPAA makes little contribution to that of the HPAA as indicated by salivary cortisol. Here we show that low birth weight is associated with increased fasting blood glucose and impaired glucose tolerance in both male and female young adult guinea pigs aged 100 days. Fasting and mean (during IVGTT) plasma cortisol was reduced in low birth weight female adult guinea pigs, and is not vary with size at birth at this age in males. This suggests that circulating cortisol does not contribute to the impaired glycaemia associated with small size at birth in the guinea pig. Glucose tolerance was increasingly impaired in males but not females, as mean plasma cortisol increased. This is consistent with cortisol impairing glycaemia in the guinea pig as in other species, in males at least. To assess the role of cortisol in prentally programmed impairment of glycaemia directly, metyrapone or vehicle containing 24% ethanol was administered to young adult guinea pigs for 3 days. Treatment with the latter impaired fasting blood glucose and glucose tolerance in females and the latter in males compared to a previous IVGTT and this was exacerbated in low birth weight females. Metyrapone prevented this impairment of fasting glycaemia and glucose tolerance in the low birth weight adult female guinea pig and in the male guinea pig regardless of birth weight class. Neither vehicle or metyrapone altered plasma cortisol, before or during a second IVGTT. Limited numbers of animals, particularly females, limited this study however and additional investigation is required. Nevertheless this shows for the first time that inhibition of glucocorticoid synthesis in the guinea pig improves glucose control. Furthermore this suggests that the low birth weight guinea pig may be more sensitive to cortisol, have increased cortisol synthesis or reduced inactivation of cortisol in peripheral tissues, leading to increased local cortisol action. In conclusion, alterations in peripheral HPAA activity in the guinea pig due to restricted fetal growth may contribute to their prenatally programmed development of impaired glucose tolerance as young adults, but the extent of that contribution may vary with age and gender. / Thesis (Ph.D.) -- University of Adelaide, School of Paediatrics and Reproductive Health, 2008

Thesis (Ph. D.)--University of Hong Kong, 2007. / Title proper from title frame. Also available in printed format.