Il est admis que l’axe endothéline (endothélines ET-1, -2 et -3 et leurs RCPG ETAR et ETBR), participe à la progression tumorale. Alors qu’ETAR est par exemple surexprimé dans le cancer de l’ovaire, ETBR l’est dans le mélanome. Cette surexpression, ainsi que l’implication d’ETA/BR dans la carcinogenèse, font de ces RCPG une cible tumorale pertinente. En raison de leurs forte spécificité, actions cytotoxiques variées, possibilités de couplage, les anticorps monoclonaux (AcM) sont des outils de choix en diagnostic et thérapie anti-cancéreuse. Cependant, on déplore actuellement l’absence d’AcM ciblant des RCPG sur le marché. Par une technique d’immunisation génique, 4 AcM anti-ETAR et 24 anti-ETBR ont été produits. Les résultats préliminaires obtenus avec les anti-ETAR sont prometteurs puisque ces AcM lient avec une haute affinité ETAR surexprimé dans des cellules CHO, l’un d’eux inhibant fortement la liaison du ligand. Mon travail de thèse s’est cependant concentré sur la caractérisation d’un anti-ETBR. Cet AcM reconnaît de façon spécifique et avec une forte affinité la conformation native d’ETBR surexprimé à la surface de cellules de mélanomes, suggérant l’existence d’une forme tumorale du récepteur. Suite à sa liaison aux cellules UACC-257 (lignée de mélanome), l’AcM se trouve internalisé. Dans ces cellules, malgré son incapacité à inhiber la liaison de l’ET, cet AcM inhibe l’activation de la voie PLC induite par le ligand et est également un fort inhibiteur de la migration due à l’activation de l’axe endothéline. Ces travaux soulignent l’intérêt de cet AcM comme outil diagnostique et thérapeutique dans le cas du mélanome. / It has been admitted that endothelin axis (endothelins ET-1, -2 and -3 and related GPCRs ETAR and ETBR) is involved in tumor progression. For instance, while ETAR is overexpressed in ovarian cancer, ETBR is in melanoma. This overexpression, as well as ETA/BR involvement in carcinogenesis, make these GPCRs a relevant tumor target. Because of their high specificity, various cytotoxic actions, possibilities of coupling, the monoclonal antibodies (mAbs) are useful tools in diagnosis and anti-cancer therapy. However, the absence of mAbs targeting GPCRs on the market is regrettable. Thanks to DNA immunization, 4 anti-ETAR mAbs and 24 anti-ETBR mAbs were produced. Preliminary results obtained with anti-ETAR are promising since these mAbs bind ETAR overexpressed in CHO cells with high affinity, one of them being a potent inhibitor of ligand binding. However, the aim of my PhD research works focused on the characterization of one anti-ETBR. This mAb specifically recognizes with high affinity the native conformation of ETBR overexpressed on the surface of melanoma cells, suggesting the existence of a tumor-specific receptor. Following its binding on UACC-257 cells (melanoma cell line), the mAb is internalized. In these cells, despite its inability to inhibit ET binding, this mAb is able to inhibit the ligand-induced activation of PLC pathway and display a potent inhibition of endothelin axis-induced migration. This work highlights the interest of this mAb as a tool for diagnosis and therapy in melanoma.

Cette thèse présente la conception d'un microgyromètre MEMS triaxial. Les microgyromètres ont de nombreuses applications telles que le contrôle d'attitude de drones ou l'interfaces homme/machine. Les microgyromètres triaxiaux sont particulièrement avantageux car ils permettent de déterminer les trois composantes de la vitesse de rotation à partir d'un seule structure monolithique et planaire. Le principe de fonctionnement des gyromètres vibrants à effet Coriolis (CVG) a été étudié analytiquement, puis une structure originale de gyromètre triaxial monolithique et planaire a été conçue. Cette structure est constituée de quatre poutres encastrées sur un cadre déformable. Des prototypes en silicium ont été réalisés et caractérisés. L’arséniure de gallium (GaAs) a été sélectionné pour la réalisation en raison de ses propriétés piézoélectriques et de son fort potentiel de miniaturisation. Un système d’électrodes pour l'excitation et la détection des vibrations mécaniques a été mis au point. Deux procédés d'usinage du GaAs ont été développés, un procédé de gravure chimique et un procédé de gravure plasma permettant tous les deux de graver verticalement le GaAs sur 450 micromètres de profondeur. Le procédé de gravure plasma est compatible avec la réalisation du CVG triaxial. Des résonateurs de test en GaAs dopé Carbone ont été réalisés par gravure chimique pour mesurer l'évolution en température de la résistivité et des propriétés électromécaniques de ce matériau. Ces mesures ont permis d'estimer que les marches aléatoires angulaires du CVG triaxial sont inférieures à 0,025 degré par racine d'heure sur la gamme de température [-40°C +80°C] pour les trois axes de mesure. Ceci situe le potentiel du CVG triaxial conçu parmi les CVG MEMS les plus performants. / This PhD present the conception of a triaxial MEMS microgyro. Microgyros offer a wide range of applications varying from drones attitude control to human interface devices. The triaxial microgyros offer great benefits because they allow determination of the three rotation rate components with only one monolithic planar structure. The operating principle of Coriolis Vibrating Gyro (CVG) has been studied analytically and an original structure has been designed. This structure consists of four beam clamped into a deformable frame. Some silicon prototypes have been machined and characterised. The gallium arsenide (GaAs) has been chosen for the realisation because of its piezoelectric properties and its great miniaturization potential. A transduction system based on GaAs piezoelectricity was developed. Two GaAs machining processes have been developed: a chemical etching process and a plasma etching process which both enable 450 micrometers deep vertical etching. The plasma etching technique allows high fidelity enough machining to be compatible with the triaxial CVG realisation. Some C-doped GaAs test resonators have been realised to measure the resistivity temperature dependency and electromechanical properties of this material. Those characterisations lead to estimate the angular random walk for the three axis ranges below 0,025 degree per square root hour on the temperature range [-40°C +80°C]. This sets the triaxial CVG together with the best monoaxial MEMS CVG.

It is believed that structures and actuation systems should be tightly integrated together in the future to create fast moving, efficient, lightweight dynamic machines. Such actuated structures could be used for morphing aircraft wings, lightweight actuated space structures, or in robotics. This requires actuators to be distributed through the structure. A tensegrity structure is a very promising candidate for this future integration due to its potentially excellent stiffness and strength-to-weight ratio, and the inherent advantage of being a multi-element structure into which actuators can be embedded. Development of these machines will utilise expertise in several fields, involving kinematics, dynamics, actuation and multi-axis motion control. The research presented in this thesis concerns the study of multi-axis actuated tensegrity structures. A form-finding method has been developed to find stable geometries and determine stiffness properties of the type of tensegrity structure proposed. It has been shown that a tensegrity structure, with practical nodes of finite size, can be designed with actuated members to give shape-changing properties while potentially allowing a good stiffness to mass ratio. An antagonistic multi-axis control scheme has been developed for the tensegrity structure. The describing function technique has been used to analyse the dead band controller in the control scheme, giving a stability criterion. An experimental actuated tensegrity system has been designed and built incorporating pneumatic muscles controlled by switching valves. Mathematical models for the experimental actuated tensegrity system have been developed in detail, including the pneumatic actuation system and the structure geometry. The dynamic behaviour of the tensegrity system has been investigated via several simulation studies, using the developed models and the proposed control scheme. Experimental validation has been successfully conducted. The multi-axis control scheme can accurately control the tensegrity structure to achieve shape changes while maintaining a desired level of internal pre-load. The mathematical models can be used as a basis for further development.

A melatonina produzida pela glândula pineal apresenta diversas ações reguladoras da homeostase dinâmica interna de mamíferos. Suas ações abrangem tanto a regulação de funções endógenas circadianas e sazonais em situações de higidez quanto durante processos fisiopatológicos. O presente trabalho avalia o efeito dos moduladores de processos inflamatórios corticosterona, TNF e IFNy; sobre o metabolismo da glândula pineal. Os resultados aqui apresentados mostram que: 1- Corticosterona potencia a síntese de noradrenalina in vivo e in vitro na vigência de estimulação β-adrenérgica, mas inibe a produção induzida por estimulação concomitante α e β-adrenérgica. 2- Este efeito é dependente de ativação de GR e não altera a captação extraneuronal de catecolaminas. 3- Corticosterona aumenta a expressão do transcrito aa-nat e a atividade das enzimas AA-NAT e HIOMT. 4 Corticosterona inibe o acúmulo nuclear de NF-κB. 5- A inibição farmacológica da via NF-κB mimetiza o efeito potenciador da corticosterona sobre a produção hormonal da pineal. 6 IFN-y inibe a via NF-κB. 7 - IFN-y potencia a produção de melatonina pela glândula pineal. 8- TNF ativa a via NF-κB. 9 TNF inibe a produção de melatonina pela pineal. 10- TNF inibe transitoriamente a expressão do transcrito aa-nat e NAS. 11 A transitoriedade deste efeito é dependente de neosíntese protéica. O presente trabalho mostra que a glândula pineal está aparelhada para responder a diferentes agentes moduladores de processos inflamatórios além de fortalecer e darem base para a hipótese da existência de um eixo imune-pineal central no controle de processos patológicos em mamíferos. / The melatonin, synthetized by the pineal gland, exihibits a number of regulatory activities on the internal dynamic homeostasis of mammals. These functions are related to the regulation of endogenous circadian and seasonal rhythms during healthy and physiopathological processes. This study evaluates the effects of the inflammatory modulators, corticosterone, TNF and IFN- y, over the pineal gland metabolism. The results presented here show that: 1 - Corticosterone enhances the synthesis of norepinephrine in vivo and in vitro in the presence of β-adrenergic stimulation, but inhibits the hormonal production when both, αand βadrenoceptors, are activated. 2 - This effect depends on the GR activation and does not interfere in the extraneuronal uptake of catecholamines. 3 - Corticosterone increases the expression of the aanat transcript and also the enzymatic activity of AA-NAT and HIOMT. 4 - Corticosterone inhibits the nuclear accumulation of NF-κB. 5 - The pharmacological inhibition of NF-κB pathway mimics the effect of corticosterone on the hormonal production in pineal. 6 - IFN-y inhibits the NF- κB pathway. 7 - IFN-y enhances the synthesis of melatonin in the pineal gland. 8- TNF activates the NF-κB pathway. 9 - TNF inhibits the production of melatonin in the pineal gland. 10 - TNF transiently inhibits the production of the aa-nat transcript and also of NAS. 11 - This transient effect depends on the synthesis of proteins. This study shows that the pineal gland can respond to different inflammatory modulators, supporting the hypothesis of a central immune-pineal axis controlling pathological processes in mammals. .

Submitted by Kenia Bernini (kenia.bernini@ufpel.edu.br) on 2017-11-06T18:31:11Z No. of bitstreams: 3 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Leonardo_Nogueira_Zanchetta_Dissertação.pdf: 1646868 bytes, checksum: f01a0b2f63bc8e6fe11ee841001804bc (MD5) Leonardo_Nogueira_Zanchetta_Produto.pdf: 616009 bytes, checksum: f2d5015935e2a6678911cef2e4daebbc (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-01-02T13:14:27Z (GMT) No. of bitstreams: 3 Leonardo_Nogueira_Zanchetta_Dissertação.pdf: 1646868 bytes, checksum: f01a0b2f63bc8e6fe11ee841001804bc (MD5) Leonardo_Nogueira_Zanchetta_Produto.pdf: 616009 bytes, checksum: f2d5015935e2a6678911cef2e4daebbc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-01-02T13:14:37Z (GMT) No. of bitstreams: 3 Leonardo_Nogueira_Zanchetta_Dissertação.pdf: 1646868 bytes, checksum: f01a0b2f63bc8e6fe11ee841001804bc (MD5) Leonardo_Nogueira_Zanchetta_Produto.pdf: 616009 bytes, checksum: f2d5015935e2a6678911cef2e4daebbc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-01-02T13:14:46Z (GMT). No. of bitstreams: 3 Leonardo_Nogueira_Zanchetta_Dissertação.pdf: 1646868 bytes, checksum: f01a0b2f63bc8e6fe11ee841001804bc (MD5) Leonardo_Nogueira_Zanchetta_Produto.pdf: 616009 bytes, checksum: f2d5015935e2a6678911cef2e4daebbc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-17 / Uma das principais questões do ensino em geral é a dificuldade em relacionar os conhecimentos. A disciplina de Biologia possui vários conteúdos, como a Microbiologia, Zoologia e a Botânica, tornando-se um desafio para o professor relacionar os conteúdos dentro de um enfoque que os unifique e dê sentido. Esta dissertação analisou o uso da Evolução como Eixo Integrador de alguns conteúdos de Biologia, no caso a Genética, Ecologia e a Evolução. Para tal foi criada uma Unidade Didática com um conjunto de 5 aulas mais um fechamento buscando esta integração entre estes conteúdos dentro de um enfoque evolutivo. As aulas abordaram os seguintes tópicos: “Aula 1 – Conceitos”, “Aula 2 - Mudança na Frequência dos Alelos, Genótipos e Fenótipos de uma população”, “Aula 3 - Tentilhões nas ilhas de Galápagos”, “Aula 4 – Impactos na Teia Alimentar” e “Aula 5 – Mutações”. A abordagem metodológica foi qualitativa, desenvolvida através de um estudo de caso para registro da aplicação da Unidade Didática com alunos do 3o ano do Ensino Médio de uma Escola Estadual. Os dados, após coletados, foram analisados por meio da análise descritiva e inferencial. Os resultados demostraram ser possível trabalhar os conteúdos de biologia utilizando a evolução como eixo integrador e que este tipo de enfoque facilita o entendimento dos conteúdos pelos alunos. Este estudo não se esgota em si mesmo, mas propõe que outros estudos ampliem as possibilidades e avancem na análise da evolução como eixo integrador para o Ensino de Biologia / One of the main current problems of teaching is the difficulty of relating knowledge. The discipline of Biology has several contents, such as Microbiology, Zoology and Botany, and it is a challenge to relate the contents within an approach that unifies and give them a logical sense. This dissertation analyzed the use of Evolution as an Integrative Axis of some Biology contents such as Genetics, Ecology and Evolution. To achieve that, a Didactic Unit was created with a set of 5 classes plus a ending task, seeking this integration between these contents within an evolutionary approach. The classes covered the following topics: "Class 1 - Concepts", "Class 2 - Change in Frequency of Alleles, Genotypes and Phenotypes within Populations", "Class 3 - Finches in the Galapagos Islands", "Class 4 - Impacts in the Food web" and "Class 5 - Mutations ". Regarding the methodology aspects, this dissertation had a qualitative approach using a Case Study of the application of the unit. The Didatic Unit was applied with students of the High School's 3rd year and a descriptive analysis of the collected data was performed. The results showed that it is possible to teach the biology contents using evolution as an integrating axis and that this type of approach facilitates the understanding of the contents by the students although still more studies are needed to develop more complete and efficient ways of working evolution as Integrating Axis

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-09T14:23:37Z No. of bitstreams: 1 Jomel Francisco dos Santos.pdf: 1033055 bytes, checksum: 0882131c9f37caac6cc396d21eb8c175 (MD5) / Made available in DSpace on 2017-02-09T14:23:37Z (GMT). No. of bitstreams: 1 Jomel Francisco dos Santos.pdf: 1033055 bytes, checksum: 0882131c9f37caac6cc396d21eb8c175 (MD5) Previous issue date: 2013-06-03 / Joint contractures are a severe complication of joint injuries that can permanently limit the extremity function. Current treatments for contractures in humans have little effect and there is an obvious need for clarification of the pathophysiology of this chronic and disabling dysfunction. Sheep joint is a promising model for the investigation of normal and pathologic states by the similarity with the biomechanics of certain human joints. The aim of this study was to investigate by the techniques of immunohistochemistry and cytochemistry the presence of key components for the development of joint contractures in sheep and to evaluate the potential of this species as an experimental model to study this disease. Therefore, we used 15 knee joint capsules of healthy Santa Inês ewes to identify myofibroblasts and mast cells. For investigation of myofibroblasts was performed immunohistochemical technique. Following histologic routine preparation, the antigen retrieval was performed by heating in citrate, followed by peroxidases blockage, blocking nonspecific protein and then primary antibody was incubated. After the secondary antibody was added to the cuts, and DAB chromogen was added. Slides were counterstained with Harris Hematoxylin and mounted. In cytochemical technique was applied the stain Toluidine Blue to disclosure mast cells. Analysis of the cuts was made under a light microscope. Positive controls of myofibroblasts (ovine cervix) and mast cells (umbilical cord) were stained satisfactorily by the standard techniques. In joint capsules, α-SMA protein was observed in the wall of arteries and rare myofibroblasts were observed in each section, as well as few mast cells were scored. Other studies about ovine joint capsule injuries must be conducted to confirm the presence of myofibroblasts and the development of joint contracture. / As contraturas articulares são complicações severas de doenças articulares que podem limitar permanentemente a função de extremidades. Os tratamentos atuais para contratura em seres humanos têm pouca eficácia e há uma evidente necessidade de esclarecimento sobre a patofisiologia dessa disfunção crônica e incapacitante. A articulação do ovino é um modelo promissor para a investigação dos estados normais e patológicos, pela semelhança com a biomecânica de determinadas articulações humanas. O objetivo deste trabalho foi investigar por meio das técnicas de imuno-histoquímica e citoquímica a presença de componentes-chave para o desenvolvimento da contratura articular na espécie ovina e avaliar a potencialidade desta espécie como modelo experimental para estudo desta patologia. Para tanto, foram utilizadas 15 cápsulas articulares de joelhos de ovelhas Santa Inês sadias para localização de miofibroblastos e mastócitos. Para investigação de miofibroblastos foi realizada a técnica de imuno-histoquímica. Após a preparação para rotina histológica, a recuperação antigênica foi realizada por aquecimento em citrato, seguida pelo bloqueio das peroxidades, bloqueio de proteínas inespecíficas e então, o anticorpo primário foi incubado. Após, o anticorpo secundário foi acrescentado aos cortes, e o cromógeno DAB foi adicionado. As lâminas foram contracoradas com Hematoxilina de Harris e montadas. Na técnica citoquímica, foi aplicada a coloração de Azul de Toluidina para evidenciação dos mastócitos. As análises dos cortes foram feitas em microscópio de luz. Os controles positivos da pesquisa de α-SMA (cérvix ovina) e de mastócitos (cordão umbilical) foram marcados ou corados satisfatoriamente pelas respectivas técnicas. Nas cápsulas articulares, a proteína foi observada na parede de artérias e raros miofibroblastos foram observados em cada corte, assim como poucos mastócitos foram corados. Outros estudos sobre lesões de cápsula articular de ovinos devem ser conduzidos para confirmar a presença de miofibroblastos e o desenvolvimento da contratura articular.

La electrocardiografía es una herramienta básica diagnóstica en la medicina veterinaria de animales de compañía, siendo la mejor alternativa para evaluar la actividad eléctrica del corazón en forma no invasiva. Sin embargo, cambios en el posicionamiento del paciente pueden ocasionar cambios en las medidas del complejo PQRST y desencadenar diagnósticos errados. El propósito de este estudio fue detectar posibles variaciones de la dirección del eje eléctrico medio cuando el registro se hace en posición decúbito lateral izquierdo en lugar de la posición estándar en decúbito lateral derecho, para analizar la variación en los resultados de acuerdo al posicionamiento del paciente. El estudio se realizó en 100 canes aparentemente sanos de ambos sexos, mayores a 1 año de edad y de cualquier peso. Se registraron dos electrocardiogramas por cada paciente: decúbito lateral derecho e izquierdo, que luego fueron analizados uno por uno; los resultados del cálculo del eje eléctrico medio fueron comparados, llegando a la conclusión que no son similares y que la posición decúbito lateral derecho es irreemplazable para el cálculo del eje eléctrico medio cardiaco en caninos.Electrocardiography is a basic diagnostic tool in veterinary medicine pet, being the best non-invasively alternative to assess the electrical activity of the heart. However, changes in patient positioning may cause changes in measures of PQRST complex and trigger misdiagnoses. The purpose of this study was to detect possible changes in the direction of the mean electrical axis when the register is in left lateral position (LL) instead of the standard right lateral position (RL), to analyze the variation in results according to patient positioning. The study was conducted in 100 apparently healthy male and female dogs, older than 1 year of age and of any weight. Two electrocardiograms were registered for each patient: right and left lateral decubitus, which were then analyzed one by one; two electrocardiograms were performed for each patient the results of the calculation of the average electrical axis were compared, concluding that they are not similar and that the right lateral decubitus position is irreplaceable for calculating mean electrical axis of the heart in dogs.

INTRODUÇÃO: A exposição ao estresse precoce (EP) pode estar associada à depressão na vida adulta. Evidencias demonstram que alterações na capacidade regulatória do eixo hipotálamo-pituitária-adrenal (HPA) sejam subjacentes a essa associação. Pacientes depressivos com EP tendem a apresentar quadros clinicamente mais graves, com pior prognóstico e resposta limitada aos tratamentos usuais. Não se sabe ainda como é o perfil cognitivo desses pacientes e como as alterações na atividade do eixo HPA impactam no desempenho neuropsicológico. OBJETIVO: O objetivo deste trabalho foi investigar a atividade do eixo HPA, através dos níveis de cortisol basal, e as funções cognitivas em pacientes depressivos com história de estresse precoce. METODO: Integrou a amostra total do estudo 107 sujeitos, sendo 77 pacientes depressivos e 30 sujeitos saudáveis com idade entre 21 e 60 anos de ambos os sexos. Foram incluídos pacientes com diagnóstico confirmado de episódio depressivo maior e gravidade de sintomas pelo menos moderada no momento das avaliações. O Questionário de Traumas na Infância (CTQ) avaliou a história de EP dividindo a amostra de pacientes em dois grupos, um com estresse precoce (EP+) e outro sem estresse precoce (EP-). Os participantes foram avaliados quanto a gravidade de sintomas psiquiátricos relacionados ao quadro depressivo, quanto à impulsividade e ao temperamento afetivo. A avaliação neuropsicológica incluiu testes de memória verbal, memória visuoespacial, memória de trabalho, atenção sustentada e dividida, além de medidas de controle inibitório, flexibilidade cognitiva, fluência verbal e QI. Na avaliação endócrina cinco amostras de cortisol salivar e de uma amostra de cortisol plasmático foram analisadas para a avaliação da atividade do eixo HPA. RESULTADOS: Setenta e dois por cento dos pacientes depressivos apresentaram EP. A história de EP influenciou o início mais precoce dos quadros depressivos (p=0,03). Não foram encontradas diferenças entre os grupos de pacientes na gravidade de sintomas psiquiátricos, comorbidades clínicas. Não encontramos diferenças entre os pacientes EP+ e EP- comparados a controles no temperamento hipertímico. Na avaliação da atividade do eixo HPA, o grupo EP+ apresentou perda do ritmo circadiano (RC) de cortisol em relação ao grupo controle, além de aumento dos níveis de cortisol salivar às 22h em comparação ao grupo EP- (p=0,04) e uma tendência comparado ao grupo controle (p=0,06). No desempenho neuropsicológico, os pacientes EP+ apresentaram prejuízos em relação aos controles em todos os subdomínios cognitivos avaliados (p<0,05 para todos os escores), exceto memória visuoespacial (p=0,13). Em contraste, os pacientes EP- mostraram déficits apenas em memória de trabalho (p=0,006), alternância atencional (p=0,01) e controle inibitório (p=0,004) comparados aos controles. Na comparação entre os grupos de pacientes, os EP+ apresentaram déficit na nomeação de cores (p=0,01) e uma tendência de prejuízo na memória verbal tardia (p=0,07). Entre os pacientes EP+ encontramos correlações moderadas entre a diminuição da variabilidade nos níveis de cortisol salivar no RC com os prejuízos na flexibilidade cognitiva (?=0,61; p=0,002) e com o controle inibitório (?=0,42; p=0,048). CONCLUSÃO: Nossos achados apontam para um perfil endócrino e neuropsicológico distinto nos pacientes EP+ em comparação com pacientes com depressão EP- e controles. A combinação da história de EP à depressão resultou em inicio mais precoce da doença, prejuízos cognitivos abrangentes e perda na manutenção do ritmo circadiano de cortisol. / BACKGROUND: Exposure to early-life stress (ELS) may be associated with depression in adulthood. Evidence shows that changes in the regulatory capacity of the hypothalamicpituitary-adrenal axis (HPA) underlie association. Patients with ELS usually are more clinically ill, showing poorer prognosis and limited response to usual treatments. It is not known yet what the cognitive profile of those patients is and how changes in the HPA axis activity would impact on cognitive functioning. AIM: The aim of this study was to investigate the HPA axis activity through basal cortisol levels and cognitive functions mediated by the hippocampus and the prefrontal cortex in depressed patients with early stress history. METHOD: Study total study sample was 107 subjects, 77 depressed patients and 30 healthy subjects aged between 21 and 60 years of both sexes. Patients had diagnosis confirmed for major depressive episode with symptom severity at least moderate by the time of the evaluations. We used the Childhood Trauma Questionnaire (CTQ) to assess the ELS history splitting the patient sample into two groups, one with early life stress (ELS+) and the other without early stress (ELS-). Participants were assessed for severity of psychiatric symptoms related to depression, such as impulsivity and affective temperament. The neuropsychological evaluation included tests for verbal memory, visuospatial memory, working memory, sustained and divided attention, inhibitory control measures, cognitive flexibility, verbal fluency, and IQ. For the endocrine assessment five samples of salivary cortisol and plasma cortisol were analyzed to evaluate HPA axis functioning. RESULTS Seventy-two percent of depressive patients had ELS. ELS itself influenced earlier onset of depressive disorders in patients (p = 0.03). Most of the affective temperaments are more prominent in patients with mood disorders than health controls. Regarding the assessment of the HPA axis activity, ELS + group showed lack of cortisol circadian rhythm (CR) compared to the control group. We also found increased salivary cortisol levels at 22 pm compared to the EP- group (p = 0.04) and a trend toward the control group (p = 0.06). In neuropsychological performance, patients EP + showed deficits compared to controls in all of the cognitive subdomains evaluated (p <0.05 for all test scores) except visuospatial memory (p = 0.13). In contrast, ELS-patients showed worse performance only in working memory (p = 0.006), attentional switching (p = 0.01) and inhibitory control (p = 0.004) compared to controls. Comparisons between patient groups showed that EP + patients had a deficit in color naming (p = 0.01) and a trend toward delayed verbal memory (p = 0.07). We found moderate positive correlations for EP+ patients between decreased variation in salivary cortisol levels in the CR and impairments in cognitive flexibility (? = 0.61; p = 0.002) and also to inhibitory control ( ? = 0.42, p = 0.048). CONCLUSION: Our findings indicate a distinct endocrine and neuropsychological profile in patients ELS + compared to depressed EP-. The combination of ELS history and depression resulted in early onset of the depression symptoms, comprehensive cognitive impairment in tasks related to the CPF and hippocampus, and failure in maintaining the circadian rhythm of cortisol.

A glândula pineal regula diversos ritmos biológicos e respostas de defesa em indivíduos hígidos através da síntese noturna de melatonina. Por outro lado, é sabido que processos inflamatórios reduzem a produção deste hormônio na glândula pineal. Neste estudo utilizamos um arranjo de qPCR para investigar a expressão circadiana de 84 genes relacionados a sinalização via receptores do tipo toll e NF-&kappa;B em glândulas pineais de rato. Confirmamos ainda, a expressão de 14 proteínas em pinealócitos através de ensaios de imunocitoquímica. Nossos resultados indicam que 70 genes inflamatórios são expressos ritmicamente na glândula pineal de ratos, enquanto 7 não são expressos e outros 7 genes são expressos, mas sem ritmicidade. Grande parte dos genes examinados apresenta padrão de expressão circadiana com maior conteúdo transcricional na fase de claro, atingindo a máxima expressão no final desta fase. Após o apagar das luzes, a expressão destes genes é rapidamente reduzida. Um possível envolvimento do ritmo de glicocorticóides endógeno sobre o padrão dessa expressão gênica foi avaliado através do bloqueio de GR por mifepristona, o que induziu a regulação da expressão de 13 genes e redução do conteúdo plasmático de melatonina no ZT18. Ainda, avaliamos o efeito da ativação dos TLR1, TLR2 e TLR6 sobre a glândula pineal e observamos que zimosan e Pam3CSK4 ativam a via do fator de transcrição NF-&kappa;B e bloqueia a síntese de melatonina induzida por noradrenalina in vitro. Por fim, utilizamos o modelo de obseidade induzido por dieta hipercalórica para avaliar se o processo inflamatório de baixa intensidade regula a síntede se melatonina in vivo. Observamos que nestas condições, a dieta hipercalórica induz rápido aumento no peso corporal e redução da produção noturna de melatonina. O efeito protetor da melatonina sobre o ganho de peso induzido por dieta foi testado através da restauração da melatonina na água de beber noturna de animais expostos à dieta. Em conjunto, nossos resultados indicam que genes inflamatórios são expressos ritmicamente na glândula pineal de ratos e influenciam a produção circadiana de melatonina via reconhecimento de padrões moleculares associados à patógenos ou sinais de perigo / The pineal gland regulates several circadian rhythms as well as immune responses in healthy animals via rhythmic production of melatonin, the hormone of darkness. On the other hand, nocturnal melatonin levels are reduced in the course of inflammation. To date, it remains to be clear the mechanisms by which the immune system affects pineal melatonin synthesis. Here we used a qPCR array profiler to investigate circadian gene expression of 84 genes related to Toll-Like Receptors and Nuclear Factor kappa B signaling. We also examined the expression of 14 proteins in pinealocytes by immunocytochemistry. Our results indicate rhythmic expression of 70 inflammatory genes, while 7 genes were not expressed and 7 were expressed without rhythmicity. The overall majority of genes tested showed a pattern of expression with a cumulative diurnal increase that peaks at the light phase of ZT12 followed by a fast reduction in the expression as soon as the light is turned off. The possible involvement of endogenous glucocorticoid rhythm in the modulation of pineal\'s inflammatory gene expression were tested by blocking Glucocorticoid Receptor (GR) using mifepristone. This procedure modulated the expression of 13 genes. In addition, the blockade of GR reduced the circulating melatonin levels at ZT18. The activation of TLR1, TLR2 and TLR6 induces the nuclear translocation of NF-&kappa;B signaling and blocks noradrenaline-induced melatonin synthesis in vitro. In addition, high-fat diet feeding increases body weight and reduce the circulating melatonin levels at ZT18. The protective role of melatonin in diet-induced weight gain was also determined by giving these rats melatonin in their drinking water at night. Altogether, our results highlight that inflammatory genes are transiently expressed in the rat pineal gland and influences the daily fluctuation of melatonin synthesis

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.