The primary objective of this study was to analyze the relationship between testosterone levels and vertical jumping performance in elite men and women athletes. The secondary objective was to verify whether testosterone levels and vertical jumping performance were different in men and women athletes and if those measurements were different between different athletic groups. Seventy (22 women and 48 men) elite athletes in track and field (sprinters), handball, volleyball, and soccer competing at national and international levels participated in the study. After 10 hours of fasting and 1 day of rest, blood samples were drawn from the antecubital vein for determining testosterone levels. Vertical jumping tests consisted of counter-movement jumps conducted on a resistive platform connected to a digital timer. Resting testosterone levels in women were 9.5% of those of the men (respectively 0.62 ± 0.06 ng·ml−1 and 6.49 ± 0.37 ng·ml−1; p < 0.001). Countermovement jump performance was significantly different between women and men athletes, with women's jumping ability 86.3% of that of men (p < 0.001). A significant positive relationship was identified between testosterone levels and vertical jump performance when all data where considered (r = 0.61, p < 0.001, n = 70).

Le carcinome médullaire de la thyroïde (CMT) est une tumeur neuroendocrine rare, se développant à partir des cellules sécrétant la calcitonine. Cette tumeur survient dans un contexte familial dans un tiers des cas. Toutes les formes germinales et près de 40% des formes sporadiques sont causées par une mutation ponctuelle activatrice de l’oncogène RET, codant pour un récepteur membranaire à activité tyrosine kinase. Les événements oncogéniques à l’origine des formes sporadiques non mutées RET restent mal définis, à l’exception de mutations activatrices des oncogènes RAS découvertes récemment.Le pronostic péjoratif du CMT est essentiellement lié à un envahissement ganglionnaire précoce. A ce titre, la chirurgie initiale est souvent insuffisante et les formes métastatiques ont longtemps été considérées en impasse thérapeutique. L’avènement récent des inhibiteurs séléctifs de tyrosine kinases (ITK) a apporté un nouvel élan à la prise en charge des tumeurs réfractaires, certains d’entre eux incluant dans leur spectre d’action le récepteur RET. Mais l’optimisation de ces traitements requiert une connaissance préalable des mécanismes moléculaires sous-jacents au développement tumoral.Dans ce contexte et en nous appuyant sur une collection importante de prélèvements humains, nous avons cherché à approfondir la decription du ‘paysage génomique’ du CMT.Dans un premier temps, nous avons évalué les anomalies structurales ponctuelles et chromosomiques présentées par les CMT. Nous avons montré, par optimisation de méthodes de séquençage, que les mutations des gènes RET et RAS interviennent dans plus de 96% des cas et que ces évènements sont mutuellement exclusifs. Ces mutations permettent de distinguer plusieurs groupes d’agressivité et de réponse aux traitements par ITK. Nous avons également observé - par technique d’hybridation génomique comparative - des anomalies de grande ampleur récurrentes dans cette pathologie : les délétions du bras court du chromosome 1 et des chromosomes entiers 4 et 22 apparaissent comme étant des évènements précoces et indépendants de la tumorigenèse du CMT.Dans un second temps, nous avons déterminé - par approche de type biopuce - les profils d’expression de microARN dans les CMT. Certains de ces régulateurs post-transcriptionnels majeurs semblent liés au caractère invasif de la tumeur, et notamment les miR-21, miR-199 et miR-129. Nous avons également démontré le potentiel d’utilisation des microARN miR-21 et miR-199 en tant que biomarqueurs circulants du CMT. L’impact fonctionnel des formes précurseurs mir-21 et mir-129 a ensuite été évalué par transfection dans les modèles cellulaires TT et MZ-CRC1.Les observations ainsi obtenues offrent de nombreuses perspectives d’études. Elles permettent la définition de marqueurs tissulaires distinguant a priori les tumeurs métastatiques et/ou réfractaires aux thérapies. Enfin, elles mettent en lumière de nouvelles pistes pour la découverte de cibles thérapeutiques additionnelles dans cette pathologie. / Medullary thyroid carcinoma (MTC) is a rare neuroendocrine tumor, arising from calcitonin-secreting cells. This cancer occurs in a family context in a third of cases. All inherited forms and nearly 40% of sporadic forms are caused by activating point-mutations in the RET oncogene, coding for a tyrosine-kinase receptor. Other oncogenic events causing sporadic cases remain unclear, but activating mutations of RAS oncogenes have been discovered recently.Prognosis of MTC is essentially linked to early lymph node occurrence. Initial surgery of metastatic forms is often insufficient and patients are considered in therapeutic dead-end. The recent advent of selective tyrosine-kinase inhibitors (TKIs) has brought a new impetus to the management of refractory tumors, some of them targeting the RET receptor. Optimization of these treatments require improving knowledge of the underlying molecular mechanisms of tumor development.In this context and helped by a large collection of human specimens, we have sought to deepen the description of genomic landscape of MTC.At first, we evaluated the structural and chromosomal abnormalities presented by MTC. We showed, by optimizing sequencing methods, that RET and RAS mutations are involved in over 96% of the cases, these events are mutually exclusives. These mutations can distinguish several groups of aggressiveness and of response to TKI treatments. We also observed, by comparative genomic hybridization techniques, recurrent abnormalities such as deletion of the short arm of chromosome 1 and loss of entire chromosomes 4 and 22. These losses appear to be early events of tumorigenesis MTC.In a second step, we determined - by a microarray approach – the microRNA expression profile of MTC. Some of these post-transcriptional regulators seem related to tumor invasiveness, such as miR-21, miR-199 and miR-129. We demonstrated the potential of microRNAs miR-21 and miR-199 as circulating diagnosis biomarkers of MTC. The functional impact of the precursor forms mir-21 and mir-129 was then evaluated by transfection in TT and MZ- CRC1 cellular models.Observations obtained pave the way for a lot of new potential studies. They allow the definition of tissue biomarkers distinguishing metastatic forms or refractory patients. Finally, they highlight new pathways for the discovery of additional therapeutic targets in this disease.

Os tumores carcinóides broncopulmonares típicos (CT) são proliferações de células neuroendócrinas. Foram consideradas como adenomas e acreditava-se que não tinham potencial para disseminação hematogênica e linfática. Porém, a ocorrência de metástase linfática e hematogênica acontece em um quinto dos indivíduos acometidos por essa patologia. A variação no comportamento clínico dos carcinóides broncopulmonares torna imperativa a realização de pesquisas que visem à melhor compreensão dessa doença. É fundamental determinar a agressividade e o risco individual da ocorrência de metástase linfática e hematogênica para que se possa oferecer um tratamento individualizado para cada binômio doente-doença. A classificação atual divide os tumores carcinóides, conforme o grau histológico de malignidade em típico e atípico, agrupando as neoplasias de acordo com o índice mitótico, relação volumétrica núcleo/citoplasma, presença ou ausência de necrose, pleomorfismo nuclear e invasão vascular. Receptores celulares na superfície externa da membrana plasmática podem ser ubiquamente expressos em diversos tipos celulares ou específicos para determinada população celular. Os receptores de somatostatina são específicos de células neuroendócrinas e também são expressos nas neoplasias desta natureza. Existem 5 tipos de receptores de somatostatina (SSTR). A interação da somatostatina (SST) com seu receptor específico provoca as inibições do ciclo celular e da angiogêsese, bem como estimula a apoptose. A meia-vida plasmática da SST é breve. Análogos com menor metabolização eram necessários. Foram desenvolvidos os análogos como octreotide e lanreotide. Estes análogos foram acoplados à radionuclídeos, possibilitando aplicação em diagnóstico, estadiamento e tratamento dos tumores neuroendócrinos. O SSTR do tipo 2 possui maior afinidade pela somatostatina. A expressão imunohistoquímica de SSTR-2 em carcinóide típicos ofereceria métodos adicionais de diagnóstico e tratamento para esta doença. Com o objetivo de demonstrar a expressão de SSTR-2 em carcinóides broncopulmonares típicos, bem como verificar se existia relação entre a expressão e ocorrência de metástase linfática 62 pacientes tiveram suas amostras de tumor submetidas ao método imunohistoquímico. Verificou-se, ademais, se a expressão de SSTR-2 e o índice de expressão imunohistoquímica eram variáveis independentes preditivas do risco de metástase linfática. A relação entre expressão de SSTR-2 e variáveis clínicas pré-operatórias também foi analisada. 36 pacientes tinham tumores que expressavam SSTR-2 (58,1%), enquanto 26 doentes tinham tumores que não expressavam SSTR-2 (41,9%). Não existiu diferença estatística significante entre a expressão de SSTR-2 e a ocorrência de metástase linfática (teste exato de Fisher, p=0,529). Também não existiram diferenças estatísticas significantes nas análises multivariadas que testaram se tanto o SSTR-2 quanto o índice de imunohistoquímica eram variáveis independentes preditivas do risco de metástase linfática. Neste estudo, os CT expressaram SSTR-2. Além disso, não existiu relação entre a ocorrência de metástase linfática e a expressão de SSTR-2. Por fim, o SSTR-2 e o índice imunohistoquímico não foram variáveis independentes do risco de metástase linfática / Typical pulmonary carcinoids are neuroendocrine cells proliferations and they were former considered lung adenomas with no hematogenic or lymphatic metastatic potential. However, it is known that up to 20% of patients develop metastatic disease. It is mandatory that new studies be developed due to the variation in clinical presentation of these patients. It is also required that the individual risk of lymphatic and hematogenic metastasis be determined in order to individualize the patients treatment. Pulmonary carcinoids are classified according to hystologic grade. The current classification includes hystologic grade, presence or absence of necrosis, nuclear pleomorphism, and vascular invasion. Somatostatin receptors (SSTR) are neuroendocrine cell specific receptors and can be detected in neuroendocrine tumors as well. There are 5 SSTRs subtypes. Somatostatin is a peptide that inhibits the cell cycle and angiogenesis as well as increases the apoptosis by binding to SSTR. The use of long-acting form of octreotide (a SST analogue) has been associated with treatment (radiolabeled somatostatin analogs) and diagnosis (OctreoScan®). Encouraging results have been obtained with the use of radiolabeled somatostain analogs yttrium-90 and Lu-177 to treat patients with neuroendocrine tumors. This study was designed to evaluate if typical bronchopulmonary carcinoid expressed Somatostatin receptor type 2 using the the immunohistochemical technique to identify Somatostatin receptor type 2. This study verified if there was relation between Somatostatin receptor type 2 expression and lymphatic metastasis. Futhermore, we verified if Somatostatin receptor type 2 and imunnohistochemistry score would be independent preditive markers to lymphatic metastasis. 62 patients were evaluated. 36 (58,1%) patients expressed Somatostatin receptor type 2 in their tumor samples whereas 26 (41,9%) patients did not express Somatostatin receptor type 2. This study did not verify significant statistical difference between SSTR-2 expression and lymphatic metastasis. Somatostatin receptor type 2 and imunnohistochemistry score were not independent preditive markers to lymphatic metastasis. There were no significant statistical differences on multivariate analyses. In conclusion this study verified that there was Somatostatin receptor type 2 expression on tumor samples studied but there was no relation between Somatostatin receptor type 2 and lymphatic metastasis. Futhermore, Somatostatin receptor type 2 and its imunnohistochemistry score were not independent preditive markers to lymphatic metastasis

Os tumores carcinóides broncopulmonares típicos (CT) são proliferações de células neuroendócrinas. Foram consideradas como adenomas e acreditava-se que não tinham potencial para disseminação hematogênica e linfática. Porém, a ocorrência de metástase linfática e hematogênica acontece em um quinto dos indivíduos acometidos por essa patologia. A variação no comportamento clínico dos carcinóides broncopulmonares torna imperativa a realização de pesquisas que visem à melhor compreensão dessa doença. É fundamental determinar a agressividade e o risco individual da ocorrência de metástase linfática e hematogênica para que se possa oferecer um tratamento individualizado para cada binômio doente-doença. A classificação atual divide os tumores carcinóides, conforme o grau histológico de malignidade em típico e atípico, agrupando as neoplasias de acordo com o índice mitótico, relação volumétrica núcleo/citoplasma, presença ou ausência de necrose, pleomorfismo nuclear e invasão vascular. Receptores celulares na superfície externa da membrana plasmática podem ser ubiquamente expressos em diversos tipos celulares ou específicos para determinada população celular. Os receptores de somatostatina são específicos de células neuroendócrinas e também são expressos nas neoplasias desta natureza. Existem 5 tipos de receptores de somatostatina (SSTR). A interação da somatostatina (SST) com seu receptor específico provoca as inibições do ciclo celular e da angiogêsese, bem como estimula a apoptose. A meia-vida plasmática da SST é breve. Análogos com menor metabolização eram necessários. Foram desenvolvidos os análogos como octreotide e lanreotide. Estes análogos foram acoplados à radionuclídeos, possibilitando aplicação em diagnóstico, estadiamento e tratamento dos tumores neuroendócrinos. O SSTR do tipo 2 possui maior afinidade pela somatostatina. A expressão imunohistoquímica de SSTR-2 em carcinóide típicos ofereceria métodos adicionais de diagnóstico e tratamento para esta doença. Com o objetivo de demonstrar a expressão de SSTR-2 em carcinóides broncopulmonares típicos, bem como verificar se existia relação entre a expressão e ocorrência de metástase linfática 62 pacientes tiveram suas amostras de tumor submetidas ao método imunohistoquímico. Verificou-se, ademais, se a expressão de SSTR-2 e o índice de expressão imunohistoquímica eram variáveis independentes preditivas do risco de metástase linfática. A relação entre expressão de SSTR-2 e variáveis clínicas pré-operatórias também foi analisada. 36 pacientes tinham tumores que expressavam SSTR-2 (58,1%), enquanto 26 doentes tinham tumores que não expressavam SSTR-2 (41,9%). Não existiu diferença estatística significante entre a expressão de SSTR-2 e a ocorrência de metástase linfática (teste exato de Fisher, p=0,529). Também não existiram diferenças estatísticas significantes nas análises multivariadas que testaram se tanto o SSTR-2 quanto o índice de imunohistoquímica eram variáveis independentes preditivas do risco de metástase linfática. Neste estudo, os CT expressaram SSTR-2. Além disso, não existiu relação entre a ocorrência de metástase linfática e a expressão de SSTR-2. Por fim, o SSTR-2 e o índice imunohistoquímico não foram variáveis independentes do risco de metástase linfática / Typical pulmonary carcinoids are neuroendocrine cells proliferations and they were former considered lung adenomas with no hematogenic or lymphatic metastatic potential. However, it is known that up to 20% of patients develop metastatic disease. It is mandatory that new studies be developed due to the variation in clinical presentation of these patients. It is also required that the individual risk of lymphatic and hematogenic metastasis be determined in order to individualize the patients treatment. Pulmonary carcinoids are classified according to hystologic grade. The current classification includes hystologic grade, presence or absence of necrosis, nuclear pleomorphism, and vascular invasion. Somatostatin receptors (SSTR) are neuroendocrine cell specific receptors and can be detected in neuroendocrine tumors as well. There are 5 SSTRs subtypes. Somatostatin is a peptide that inhibits the cell cycle and angiogenesis as well as increases the apoptosis by binding to SSTR. The use of long-acting form of octreotide (a SST analogue) has been associated with treatment (radiolabeled somatostatin analogs) and diagnosis (OctreoScan®). Encouraging results have been obtained with the use of radiolabeled somatostain analogs yttrium-90 and Lu-177 to treat patients with neuroendocrine tumors. This study was designed to evaluate if typical bronchopulmonary carcinoid expressed Somatostatin receptor type 2 using the the immunohistochemical technique to identify Somatostatin receptor type 2. This study verified if there was relation between Somatostatin receptor type 2 expression and lymphatic metastasis. Futhermore, we verified if Somatostatin receptor type 2 and imunnohistochemistry score would be independent preditive markers to lymphatic metastasis. 62 patients were evaluated. 36 (58,1%) patients expressed Somatostatin receptor type 2 in their tumor samples whereas 26 (41,9%) patients did not express Somatostatin receptor type 2. This study did not verify significant statistical difference between SSTR-2 expression and lymphatic metastasis. Somatostatin receptor type 2 and imunnohistochemistry score were not independent preditive markers to lymphatic metastasis. There were no significant statistical differences on multivariate analyses. In conclusion this study verified that there was Somatostatin receptor type 2 expression on tumor samples studied but there was no relation between Somatostatin receptor type 2 and lymphatic metastasis. Futhermore, Somatostatin receptor type 2 and its imunnohistochemistry score were not independent preditive markers to lymphatic metastasis

Introdução: Os tumores carcinóides broncopulmonares típicos são proliferações malignas neuroendócrinas. Até bem pouco tempo eram consideradas como adenomas, isto é, tumores benignos. Porém com o avanço dos estudos anatomopatológicos, foi identificada a sua face maligna, pois apresenta as principais características das neoplasias malignas, quais sejam: metástase e invasão tecidual local. Além das metástases, estes tumores são capazes de produzir outra entidade ainda pouco estudada e conhecida que é a micrometástase. Estas correspondem a metástases menores que 2mm de diâmetro, que podem ou não se desenvolver, causando recidiva tumoral. Por sua vez as micrometástases são divididas em grupos de células tumorais, com diâmetro de 0,2 a 2mm e células tumorais isoladas, com diâmetro menor do que 0,2 mm. A literatura nos mostra que a incidência de micrometástase varia entre 10 a 90% dos pacientes em diversos tumores estudados. No caso dos carcinóides típicos temos pouca informação a respeito, sendo que a literatura nos mostra que a micrometástase em tumores carcinóides é considerada com fator de pior prognóstico. Porém não é o que observamos clinicamente, uma vez que temos o seguimento de inúmeros pacientes por mais de 10 anos, sem a recidiva tumoral em linfonodos mediastinais (seguimento clínicoradiológico). Objetivos: Verificar a presença de micrometástases em suas diversas formas, em pacientes portadores de carcinóide típico broncopulmonar, e verificar a possibilidade da predição do risco individual destas micrometástases em função de variáveis clínicas, anatomopatológicas e biomarcadores teciduais. Casuística e Métodos: Quarenta e nove pacientes portadores de carcinóide típico broncopulmonar com acompanhamento mínimo de 5 anos foram estudados. Todos foram submetidos a ressecção linfonodal por amostragem ou radical. As seguintes variáveis foram coletadas dos prontuários ou por entrevista: gênero, idade, localização do tumor em relação à carina (central ou periférico), diâmetro da lesão, comprometimento da margem cirúrgica, estadiamento TNM, ocorrência de metástases linfonodais, bem como quantidade de linfonodos acometidos por neoplasia em relação ao total dissecado, metástases à distância e tempo de sobrevivência. Os linfonodos foram analisados por coloração de hematoxilina-eosina e por imuno-histoquímica (Sinaptofisina e Cromogranina A) para pesquisa de micrometástase. Resultados: O grupo foi composto por 19 homens (38,8%) e 30 mulheres (61,2%). A idade média dos pacientes foi de 41,3 anos. Houve uma distribuição regular entre todos os lobos pulmonares acometidos. Em relação às vias aéreas, 78% dos tumores eram centrais e 22% eram periféricos. O diâmetro do maior eixo do tumor primário dos 49 pacientes variou de cinco a 80 milímetros, com mediana de 25 e intervalo interquartil 25 a 75% entre 16 e 35 milímetros. Em 54% dos casos foi realizada lobectomia pulmonar, 18% pneumonectomia, 12% bilobectomias e 16% procedimentos poupadores (segmentectomias, broncoplastias e nodulectomias). Em 12% dos casos houve comprometimento da margem cirúrgica. Em 42,8% dos casos houve imunomarcação por pelo menos um dos biomarcadores Sinaptofisina ou Cromogranina A para micrometástase. Em 18,4% dos casos foi diagnosticada macrometástase linfática, e em 1 caso ocorreu metástase hematogênica. Foram realizadas 4 baterias de testes avaliando os grupos sem e com metástases/micrometástases para se verificar a possibilidade de predição do risco individual de micrometástase. Conclusão: Foi possível encontrar micrometástases linfáticas utilizando imuno-histoquímica (Sinaptofisina e Cromogranina A). Não foi possível predizer o risco individual de micrometástases nos grupos estudados. Não houve diferença entre os grupos sem e com qualquer tipo de micrometástase. Não foi possível estabelecer correlação entre incidência de metástase e micrometástase nesta amostra populacional. / Introduction: The typical lung carcinoids are neuroendocrine tumors. Until short time ago they were considered adenomas, that is, benign tumors. Although, due to the anatomopathologic advances, it was identified its malignant behavior, once it presents the main characteristics of the malignant tumors: matastasis and local invasion. Beyond the metastasis, this tumor is able to produce another entity not yet well studied, the micrometastasis. This corresponds to metastasis shorter than 2mm in diameter that can or not develop and cause tumoral recurrence. The micrometastasis are divided in two groups: clusters, with diameter between 0,2 and 2mm, and isolated tumor cells, with diameter less than 0,2mm. The medical literature shows that the incidence of micrometastasis of different tumors has a wide variation, between 10 to 90%. In the case of the typical lung carcinoids few information is presented, and the presence of the micrometastasis worsen prognosis. On the other hand this is not what we usually see clinically, once the follow up of numerous patients of our casuistic for more than 10 years did not show the recurrence of the desease in the mediastinal lymphnodes. Objectives: Verify the presence of micrometastasis in its various forms in patients comited by lung carcinoid tumors and verify the possibility to predict the individual risk of micrometastasis from clinical and anatomopathological variables and tissue biomarkers. Casuistic and Methods: Forty nine patients with lung carcinoid tumors with follow up of at least 5 years were studied. All of them were submitted to mediastinal lymphnode dissection during the surgical procedure. The data collected was: age, gender, tumor location (central or peripherical), diameter, compromised surgical edge, TNM stage, lymphatic metastasis, hematogenic metastasis and survive. The lymphnodes were analised by Hematoxilin-Eosin and immunohistochemistry (Synaptophysin and Chromogranin A) in order to search for micrometastasis. Results: There were 19 men (38,8%) and 30 women (61,2%) with a mean age of 41,3 years. There was a regular distribution in all pulmonary lobes. There were 78% of central and 22% periferic tumors. The diameter varied between 0,5 to 80mm, with median in 25. In 54% of the cases was performed pulmonary lobectomy, in 18% pneumonectomy, in 12% bilobectomy and in 16% other procedures (bronchoplasty, wedge resection, nodulectomy). In 42,8% there was immunostaining with Synatophysin or Chromogranin A to micrometastasis. In 18,4% was diagnosed macrometastasis and in 1 case there was haematogenic metastasis. It was done 4 batteries of statistical tests to verify the possibility of prediction of the individual risk of micrometastasis. Conclusion: It was possible to find lymphatic micrometastasis using immunostaining with Synaptophysin and Chromogranin A. It was not possible to predict the individual risk of micrometastasis in the studied groups. There was no difference between the groups with or without micrometastasis. It was not possible to estabilish a correlation between the incidence of macro and micrometastasis in this population.

Les carcinoïdes pulmonaires, les carcinomes neuroendocriniens à grandes cellules (LCNEC) et les mésothéliomes malins sont des tumeurs thoraciques rares, dont l'incidence a augmenté au cours des dernières années. Le diagnostic de ces tumeurs est soumis à la variabilité inter-observateur et les opportunités thérapeutiques sont limitées. De grandes études génomiques visant à les caractériser au niveau moléculaire pourraient aider à mieux comprendre les mécanismes sous-jacents à leur développement et faciliter le diagnostic et le traitement de ces maladies. Mon projet de thèse visait à combler les lacunes dans la compréhension des carcinoïdes pulmonaires, des LCNEC et du mésothéliome péritonéal malin. À la suite des travaux entrepris au cours de ma thèse, nous avons constaté que (1) de la même manière que le mésothéliome pleural, les mésothéliomes péritonéaux sont également caractérisés par des mutations conduisant à la perte d'expression de BAP1, facteur de bon pronostic, (2) les patients atteints d’un LCNEC conservant une expression de RB1 présentent de meilleurs résultats lorsqu’ils sont traité avec une chimiothérapie du cancer du poumon non à petites cellules par rapport à une chimiothérapie du cancer du poumon à petites cellules, (3) les carcinoïdes pulmonaires peuvent être classés en trois groupes moléculaires pertinents sur le plan clinique, et (4) , l'identification de supra carcinoïdes confirme l'existence d'un lien moléculaire entre les néoplasmes neuroendocriniens pulmonaires de faible et de haut grade / Pulmonary carcinoids, large-cell neuroendocrine carcinomas (LCNEC), and malignant mesotheliomas are rare thoracic tumours, which incidence has been increasing over the past years. The diagnosis of these tumours is subjected to inter-observer variability and the therapeutic opportunities are limited. Large genomic studies to characterize them at a molecular level might help to better understand the mechanisms underlying their development, and to help the diagnosis and treatment of these diseases. My thesis project aimed to fill the gap in the understanding of pulmonary carcinoids, LCNEC, and malignant peritoneal mesothelioma. As result of the work undertaken during my thesis, we found that (1) similarly to pleural mesothelioma, peritoneal mesotheliomas are also characterised by mutations leading to the loss of expression of BAP1, which is a factor of good prognostic, (2) LCNEC patients with a remaining expression of RB1 have a better outcome when treated with non-small cell lung cancer chemotherapy in comparison to small-cell lung cancer chemotherapy, (3) pulmonary carcinoids can be classified in three clinically-relevant molecular groups, and (4), the identification of supra carcinoids supports a molecular link between the low and high-grade lung neuroendocrine neoplasms

Prostate cancer (PCa) is the most common cancer in American men. Although initial androgen deprivation therapy (ADT) confers a five year survival rate of 99%, the relapse of metastatic and drug resistant PCa (CRPC- Castration-Resistant PCa) continues to account for most deaths. How certain PCa cells develop into CRPC is the key question in the field. In addressing it, attention has focused on epigenetic factors that contribute to CRPC development. Herein we investigated the role and regulation of histone demethylase PHF8 during PCa neuroendocrine differentiation (NED) and progression into CRPC. We utilized bioinformatic analyses and biochemical approaches in PCa/CRPC cell line and mouse models to unravel the following results: First, we discovered that PHF8 post-transcriptionally clusters with cell cycle genes during NED and into CRPC via an AR/MYC/miR-22 regulatory axis. We showed that this axis is dysregulated in CRPC cells to allow enhanced cell proliferation and resistance to the clinical AR antagonist drug Xtandi® (enzalutamide). Second, we revealed that PHF8 is necessary for hypoxia induced NED by demethylating repressive H3K9me2 and H3K27me2, above maintaining active H3K4me3 on select NED genes. Importantly, we unveiled that PHF8 sustains HIF1α expression in CRPC cells via a regulatory role associated with full length AR. Third, we recapitulated the role of PHF8 in vivo by excising its floxed allele in the prostate of TRAMP mice -Transgenic Adenocarcinoma of the Mouse Prostate. We observed that KO of Phf8 lowered tumor burden in part by sustaining Ezh2 expression during NED transition into CRPC. In conclusion, our data implicates PHF8 in multiple oncogenic roles and regulations during PCa NED into CRPC. Our results lay a foundation for understanding the dynamics of histone modifying enzymes during PCa progression and hint at designing small molecule inhibitors against PHF8 as a novel CRPC therapeutic target.

Small intestinal neuroendocrine tumor (SI-NET) is the most common small bowel tumor in Europe and USA, with an annual incidence of around 0.3-1.3/100000 persons. SI-NETs are the most common type of gastroenteropancreatic NETs (GEP-NETs), and they are known for their ability to produce hormones such as tachykinins and serotonin, as well as for their favorable long-term prognosis in comparison to gastrointestinal adenocarcinoma. The overall aim of the thesis was to investigate unknown or unclear aspects of SI-NET disease, in connection with prognosis, treatment and follow-up. Paper I confirmed several known negative prognostic factors and also showed, for the first time, that para-aortal lymph node metastases and peritoneal carcinomatosis were associated with worse survival by multivariable analyses. Locoregional surgery was associated with a low post-operative mortality, and a prolonged long-term survival by multivariable analysis. In Paper II we continued to investigate peritoneal carcinomatosis and found it be a risk factor not only for death, but also for emergency re-surgery. Furthermore, genetic analyses of samples from primary tumors in patients with and without peritoneal carcinomatosis showed a difference in the DNA between these two groups. In Paper III the outcome after liver surgery and/or radiofrequency ablation of liver metastases was investigated. To summarize, no difference in survival was seen in patients treated with surgery/radiofrequency ablation in comparison with matched controls. However, a superior radiological response of liver metasases and lower U-5-HIAA values were seen in patients subjected to liver surgery and/or radiofrequency ablation compared to matched controls. Paper IV compared ultrasonography, computed tomography and 11C-5HTP-PET in the follow-up after radiofrequency ablation of NET liver metastases. The study concluded that 11C-5HTP-PET depicted all residual tumors after RFA and that it, if used, should be combined with computed tomography for easier interpretation, as RFA areas are not clearly distinguishable with 11C-5HTP-PET alone. Paper V studied gallstone complications after somatostatin analog treatment in SI-NET patients, and concluded that there was a rather high risk to be subjected to a cholecystectomy due to biliary colic, cholecystitis, cholangitis or pancreatitis after primary surgery in somatostatin analog treated patients.

Aquaculture is an essential developing sector for world food production however one of the major bottlenecks for the sustainability of the aquaculture industry is the ability to control fish reproduction in captivity and to produce high quality seeds. European sea bass is a one of most commercially important species for the European fish farming industry. If broodstock management under captivity is well established, problems remain in hatcheries where survival can be low and deformity prevalence high as well as in on growing sites where fish reach puberty early especially with skewed sex ratio towards males. Sea bass displays strong seasonality in its physiology and is therefore an excellent candidate for the study of the photo-neuroendocrine control of reproduction and growth. The overall aims of this thesis were to better understand the molecular and endocrine drivers that control the Brain-Pituitary-Gonad axis in repeat spawner sea bass, and expand our knowledge of sea bass light and temperature regulation of melatonin production. First, this PhD project investigated the seasonal expression of kisspeptin, GnRH and gonadotropin genes in relation to the gonadal development throughout a reproductive cycle in male repeat spawning sea bass (Chapter 3). A partial sequence for the receptor kissr4 was isolated and described showing similarity to all other teleost species sequences available to date. QPCR molecular assays were validated to mesure the expression of a suite of genes along the BPG axis including kisspeptin related genes (Kiss1 and Kiss2 and its receptor kissr4) over a full reproductive cycle (12 months) in adult male European sea bass. Brain Kisspeptin mRNA expression levels (kiss1, 2 and kissr4) showed clear seasonal profiles and correlated well to other BPG markers (GnRHs, fshβ and lhβ), supporting a possible involvement of kisspeptin genes in the seasonal control of reproduction in repeat spawning sea bass. Moreover, clear seasonal patterns were observed for expression of the genes encoding for pituitary mRNA expression of lhβ and fshβ, with a significant correlation between expression of both subunits and GSI and steroids levels. However, no clear seasonal profiles in brain GnRHs gene expression were observed with the exception to some peaks in GnRH1 and GnRH2. The second part of this PhD project investigated the potential direct effect of the two kisspeptin core peptides (kiss1 and kiss2) on the pituitary gonadotropin gene expression (Chapter 4). The aim of this work was to better understand the mechanism by which kisspeptin acts on the BPG axis. This was done by testing the kisspeptin decapeptide core sequences on the lhβ and fshβ transcript expression in primary culture of sea bass pituitary cells using QPCR technique. The findings, as a whole, provided evidence that kisspeptin can act directly on the pituitary gonadotroph cells and modulate fshß and lhß mRNA expression in sea bass although effects were limited and not uniform. Of note, kissr4 gene expression was also detected in the sea bass pituitary. The third part of this PhD project focused on the effects of environmental signals (photoperiod and temperature) on melatonin production (Chapter 5). Environmental manipulation is routinely used in the aquaculture industry with the purpose of enhancing growth and manipulating the timing of reproduction in seasonal fish species like sea bass. Melatonin, known as the light perception and time keeping hormone, has been suggested to play key roles in the synchronisation of most physiological functions in vertebrates, although the mechanisms by which melatonin controls reproduction, growth and behaviour are still not fully understood in fish. The studies performed aimed .to determine the synergistic effects of both temperature and photoperiod on the daily phase and amplitudinal changes in melatonin production through both in vivo and in vitro trials. The results confirmed the diel melatonin rhythm in sea bass as previously reported in many teleost species with “high at night” and “low at day” melatonin profiles. Temperature showed clear effects on the amplitude of the melatonin production under both in vivo and in vitro conditions for both long day and short day photoperiods. Furthermore, no endogenous melatonin production was found under constant darkness in both in vivo and in vitro conditions. These results suggested a lack of intrapineal (or located elsewhere such as retina and/or deep brain) oscillators in sea bass, contrasting with previous reports. These results further enhance our knowledge of light perception and circadian rhythmicity in sea bass, while the circadian system remains to be characterised in sea bass and teleosts as a whole. Overall, this doctoral work broadened our understanding on the photoneuroendocrine control of reproduction in a seasonal fish species, sea bass. New knowledge gained and tools developed from this work should help to develop/optimise husbandry techniques for the sea bass farming industry with the view to increase production and profitability and thus promoting the sustainable expansion of the sea bass aquaculture in Europe. It has also the potential to help the fishery sector in the modelling of wild sea bass populations.

Made available in DSpace on 2016-03-15T19:40:22Z (GMT). No. of bitstreams: 1 Maria Aparecida Bernardes Orlandi.pdf: 573960 bytes, checksum: 40c15cb8f5dd9faf220d051de6f44829 (MD5) Previous issue date: 2006-12-12 / Fundo Mackenzie de Pesquisa / The aim of this research was to identify in literature, possible variations and some relationship among aggression, neuroendocrine stress and neurocognitive deficit. A meta-analysis was conduct based on reviews, essays and thesis, on electronic database selected from 1996 to 2006, on MEDLINE, ERIC and CAPES. From a total of 371 selected researches, 41 were considered relevant, 330 were rejected because they did not satisfied the criteria of the analysis due to psychobiological context of aggression development in childhood related to the subject. The results suggested at least 7 different perspectives through childhood aggression could be analysed with favorable hypothesis, evidences and considerations related to neuroendocrine stress and neurocognitive deficit as co-occurrent with aggression in case of convergent situations of multifactorial vulnerability of the child. / O objetivo deste trabalho foi identificar na literatura, possível variação da agressividade e sua relação com o estresse neuroendócrino e déficit neurocognitivo na infância. Foi conduzida uma meta-análise baseada em artigos de revisão, dissertações e teses, selecionados nas fontes eletrônicas do MEDLINE, ERIC E CAPES, no período de 1996 a 2006. Do total de 371 artigos selecionados, 41 foram considerados relevantes, 330 foram descartados por não cumprirem os critérios para a análise devido ao contexto psicobiológico do desenvolvimento da agressividade na infância relacionado ao tema da pesquisa. Os resultados demonstraram pelo menos 7 diferentes perspectivas pelas quais a agressividade infantil pode ser analisada, com hipóteses, evidências e considerações favoráveis à relacionar o estresse neuroendócrino e déficit neurocognitivo como coocorrentes com a agressividade, em situações de convergência multifatorial e de vulnerabilidade da criança.