O sistema laguna-barreira da Feitoria localizado na margem oeste da célula Sul da laguna dos Patos – Rio Grande do Sul, Brasil – se assemelha geomorfologicamente, em uma escala menor, a um sistema barreira oceânico e cronologicamente ao sistema Barreira IV (Villwock e Tomazelli, 1995). Com o intuito de caracterizar a morfologia e a estratigrafia deste sistema deposicional laguna-barreira foi estudada a geologia de superfície e de subsuperfície. As características geológicas de superfície foram obtidas através da análise de aerofotografias e de imagens de satélite; de levantamentos topográficos e das análises geocronológicas por Luminescência Oticamente Estimulada em sedimentos superficiais. A geologia de subsuperfície foi caraterizada a partir da análise de perfis geofísicos registrados com sísmica rasa e de seções de georradar; junto foram integradas as análises granulométricas, palinológicas e geocronológicas (radioisótopos de 14C) coletadas em testemunhos de sondagem. Através da compilação crítica dos resultados obtidos, esta Tese apresenta evidências de que a gênese da barreira da Feitoria e da laguna na sua retaguarda, está correlacionada a topografia antecedente, marcada pela existência de um alto topográfico remanescente entre os paleovales fluviais dos rios Turuçu e Corrientes, onde este alto serviu de substrato para o início do desenvolvimento desta barreira. Também, é evidenciado que a evolução geomorfológica da barreira da Feitoria (orientação, truncamentos e tipologia dos cordões litorâneos), subdividida em nove fases de deposição, ocorreu associada as variações relativas do nível do mar a partir do Holoceno médio (8.000 anos A.P.). Observa-se também que foi influenciada diretamente pelas flutuações de alta frequência do nível de base da laguna dos Patos, causadas pela interação de três fatores: variabilidade no volume de precipitação na área da bacia de drenagem; marés meteorológicas causadas pelo vento; e alterações geomorfológicas na desembocadura da laguna dos Patos. O início da progradação dos cordões litorâneos na barreira da Feitoria ocorreu, aproximadamente, a 7.200 anos A.P., cronocorrelato ao período em que o nível relativo médio do mar ultrapassou o nível atual. A cota altimétrica da 1° crista destes cordões litorâneos encontra-se a 4,2 m acima do nível do mar atual, evidenciando a sobre-elevação do nível de base lagunar para a formação dos mesmos. A ilha da Feitoria, porção sul da barreira, teve sua formação, aproximadamente, a 5.000 anos A.P. associada ao rebaixamento do nível do mar e, consequentemente, ao afloramento do paleoalto topográfico da Feitoria. A partir deste período o controle sobre a morfologia e a orientação dos cordões litorâneos passa a ter uma maior influência associada as modificações na geomorfologia da desembocadura da laguna dos Patos (alta frequência). No entanto, as variações do nível de base lagunar (volume de precipitação e marés meteorológicas) de mais alta frequência são as responsáveis pela gênese das cristas dos cordões litorâneos. A deposição de sedimentos finos na área da laguna Pequena (retaguarda) começa antecedentemente a 7.730 anos A.P. com um aumento de influência marinha, próximo a esta idade, passando por um curto período com menor influência, retornando a aumentar a entrada de água salgada no interior da laguna dos Patos próximo a 7.000 anos A.P. A partir deste período, apesar das oscilações de alta frequência, passa a ter uma tendência de diminuir gradativamente a influência marinha no interior das lagunas dos Patos e Pequena. Esta diminuição da influência marinha pode ser correlacionada ao estreitamento do canal de ligação entre a laguna dos Patos e o oceano, assim como, o desenvolvimento da barreira da Feitoria diminuiu a comunicação entre a laguna Pequena e a laguna dos Patos. / The lagoon-barrier system of the Feitoria located on the western margin of the southern cell of the Patos lagoon - Rio Grande do Sul, Brazil - resembles geomorphologically, on a smaller scale, to the barrier system oceanic and chronologically to the Barrier IV system (Villwock e Tomazelli, 1995). In order to characterize the morphology and stratigraphy of this depositional lagoon-barrier system, the surface and subsurface geology were studied. The surface geological characteristics were obtained from the analysis of aerial photographs and satellite images; surveying and geochronological analyzes Optically Stimulated Luminescence of surface sediments. The subsurface geology was characterized by the analysis of geophysical profiles recorded with shallow and georadar seismic; and granulometric, palynological and geochronological analyzes (14C radioisotopes) of sedimentary records. This thesis presents evidences that the genesis of the Feitoria barrier and the Pequena lagoon is correlated with the previous topography, marked by the existence of a high topographic remnant among the fluvial paleovales of the Turuçu and Corrientes rivers, where this high served as a substrate for the beginning of the development of this barrier. Also, it is evidenced that the geomorphological evolution of the Feitoria barrier (orientation, truncations and typology of beach ridges), subdivided into 9 phases of deposition, was associated with sea level variations from the average Holocene (8,000 years Before the Present) and was also influenced directly by the high frequency fluctuations of the basal level of the Patos lagoon, caused by the interaction of three factors: variability in the volume of precipitation in the area of the drainage basin; wind tides; And geomorphological changes at the mouth of the Patos lagoon. The beginning of the development of the coastal strands in the Feitoria barrier occurred approximately 7,200 years B.P., chronologically during the period in which the average relative sea level exceeded the current level. The crest of the first ridge of these coastal strands is 4.2 m above current sea level, evidencing the over-elevation of the lagoon base level for their formation. The island of Feitoria, southern portion of the barrier, began approximately 5,000 years BP associated with the decline of sea level and the outcropping of the topographic paleoalto of Feitoria. From this period, the control over the morphology and orientation of the coastal strands will have a greater influence associated with the changes in the geomorphology of the mouth of the Patos lagoon (high frequency). However, variations in lagoon base level (volume of precipitation and wind tides) the highest frequency are responsible for the genesis of coastal beach ridges. The deposition of fine sediments in the Pequena lagoon area starts before 7,730 years BP with an increase of marine influence, near this age (palinological records), passing for a short period with less influence, returning to increase the entrance of salt water inside Patos lagoon, near to 7,000 years BP. From this period, despite the high frequency oscillations, there is a tendency gradually decrease the marine influence inside the lagoons Patos and Pequena. This decrease in the marine influence can be correlated to the narrowing of the channel connecting the Patos lagoon to the ocean, as well as the development of the Feitoria barrier, reducing the communication between the lagoons Pequena and Patos.

O glioblastoma multiforme (GBM) é uma doença grave e sem tratamento eficaz, especialmente pelos agentes terapêuticos disponíveis causarem reações adversas importantes nas doses terapêuticas. O metotrexato (MTX) é um fármaco citotóxico utilizado para tratar diversas neoplasias, no entanto, sua utilização é limitada pela baixa biodisponibilidade e reações adversas. A nanotecnologia tem sido utilizada para aumentar a eficácia dos medicamentos antitumorais, com o intuito de direcioná-los para o sítio de ação e reduzir os efeitos adversos. Nesse sentido, realizamos ensaios com nanocápsulas lipídicas de MTX (LNC MTX) para avaliar os mecanismos de captação em linhagens celulares de glioblastoma e micróglia, além investigar a eficácia terapêutica da LNC MTX em ensaios in vitro e in vivo. Inicialmente, ensaios de microscopia de fluorescência, empregando bloqueadores farmacológicos específicos para transportes de membrana, mostraram que as LNC MTX marcadas com Rodamina B penetram em células tumorais GL261 por endocitose, dependente de caveolinas, e em células de micróglia da linhagem BV2 por fagocitose e macropinocitose. Os tratamentos com LNC MTX ou solução de MTX (em concentrações correspondentes) em células GL261 inibiram a proliferação; aumentaram a fragmentação de DNA, mas, somente as LNC induziram a morte celular por necrose e diminuíram o número de células na fase G1/G0 do ciclo celular. Na linhagem celular BV2, os tratamentos com LNC MTX ou solução de MTX inibiram a proliferação, reduziram a quantidade de células na fase G1/G0 do ciclo celular, aumentaram a fragmentação de DNA e induziram morte celular por apoptose e apoptose tardia. Os ensaios in vivo de microscopia intravital mostraram que a LNC MTX atravessa a barreira Hematoencefálica (BHE) de camundongos fêmea C57Bl/6 após administração intravenosa ou oral, sem danificar a sua estrutura. O tamanho do glioblastoma in vivo foi reduzido em animais tratados com LNC MTX por via oral em relação aos animais tratados com salina. Esta redução não foi detectada em animais tratados com solução de MTX. Em conjunto, os dados obtidos mostram que a LNC MTX penetram em células de glioma e da glia e causam toxicidade, atravessam a BHE in vivo e sugerem que a nanoencapsulação do MTX pode ser uma estratégia importante para o tratamento do glioblastoma. / Glioblastoma multiforme (GBM) is a serious disease and no effective treatment is availabe, especially because the drugs cause significant adverse reactions in therapeutic doses. Methotrexate (MTX) is a cytotoxic drug used to treat many neoplasms, however, their use is limited by the low bioavailability and adverse reactions. Nanotechnology has been used to increase the effectiveness of antitumor drugs in order to direct them to the site of action and to reduce adverse effects. Accordingly, we carried out an experimental approach with MTX lipid nanocapsules (MTX LNC) to evaluate the uptake mechanisms in glioblastoma and microglia cell lines, and the therapeutic efficacy of MTX LNC in vitro and in vivo systems. Initially, fluorescence microscopy assays employing specific pharmacological blockers for membrane transport showed that the MTX LNC stained with Rhodamine B penetrated into GL261 tumor cells by caveolae-mediated endocytosis, and in BV2 microglia cells by phagocytosis and macropinocytosis. Treatment with MTX solution or MTX LNC (at corresponding concentrations) on GL261 cells inhibited the proliferation; increased DNA fragmentation, but only the LNC induced cell death by necrosis and decreased the number of cells in the G1/G0 phase of the cell cycle. In BV2 cells, treatment with MTX solution or MTX LNC inhibited proliferation, reduced number of cells in the G1/G0 phase of the cell cycle, increased DNA fragmentation and cell death, induced by apoptosis and late apoptosis. Intravital microscopy study showed that the MTX LNC across the Blood-Brain Barrier (BBB) of C57BL/6 female mice after intravenous or oral administrations, without damaging its structure. The area of glioblastoma in vivo was reduced in animals oral treated with MTX LNC comparing to saline treated mice. This reduction was not observed in animals treated with MTX solution. Together, the data herein obtained show that MTX LNC penetrate the cell membrane and cause cell toxicity on glioma and neurons lineage, cross the BBB and suggest that the nanoencapsulation of MTX can be an important strategy for the treatment of glioblastoma.

Small vessel disease (SVD) is an important cause of stroke, cognitive decline, and age-related disability. The cause of SVD is unknown, increasing evidence from neuropathology and neuroimaging suggests that failure of the blood-brain barrier (BBB) precipitates or worsens cerebral SVD progression and its failure is associated with SVD features such as white matter hyperintensities (WMH), perivascular spaces (PVS) and lacunar infarcts. The BBB change mechanism may also contribute to other common disorders of ageing such as Alzheimer's disease (AD). Magnetic resonance imaging (MRI) has revolutionised our understanding of SVD features. The MRI contributes to better understanding of the SVD pathophysiology and their clinical correlates. The purpose of this project was to better understand the pathogenesis of SVD, which involves improved understanding of BBB structures and pathophysiology and accurate measurement of cerebral SVD imaging characteristics on MRI scans. We aimed to assess (1) structures related to the BBB and factors that affect the BBB; (2) efficient and consistent WMH measurement method; (3) effect of stroke lesions on WMH and cerebral atrophy progression; (4) development and optimisation of computational PVS measurement method; (5) the relationships between PVS and SVD, blood markers, and BBB permeability. Section one describes structures and pathophysiology of the BBB. I reviewed the BBB structural and functional components from the view of neurovascular unit, PVS, and junctional proteins. The PVS part was done in a systematic search. I also reviewed some common stimuli for BBB permeability including inflammation and ischemia. Ischemic triggers for the BBB permeability were summarized systematically. Based on the literatures above, I summarized changes in junctional proteins in ischemia, inflammatory pain and AD models. Section two describes accurate measurement of WMH progression and atrophy. I used data from 100 patients who participated in a stroke study about BBB permeability changes in lacunar versus cortical stroke. To find a most efficient and consistent WMH measurement method, we tested several computational methods and effect of common processing steps including bias field correction and intensity adjustment. To avoid the effect of artefacts, I did a systematic search about artefacts and tested methods of image segmentation to avoid WMH artefacts as much as possible. To investigate the effect of stroke lesions on WMH and atrophy progression, I did the WMH, atrophy segmentation and stroke lesion measurements in a subgroup of 46 patients with follow-up scans, and showed that stroke lesions distorted measurement of WMH and atrophy progression and should be excluded. Section three describes development and optimization of a computational PVS measurement method, which measures the count and volume for PVS based on a threshold method using AnalyzeTM software. We tested the observer variability and validated it by comparison with visual rating scores. We investigated the associations between PVS results with other SVD features (WMH, atrophy), risk factors (hypertension, smoking and diabetes), blood markers, and BBB permeability. In conclusion, MRI is a valuable tool for the investigation of cerebral SVD features and BBB permeability. Exclusions of artefacts and stroke lesions are important in accurate measurement of WMH. PVS are important features of BBB abnormalities, and they correlate and share risk factors with other SVD features, and they should be considered as a marker of SVD and BBB permeability. Further systematic histological and ultrastructural studies of BBB are desirable in understanding the BBB regarding to the different parts of the cerebral vascular tree.

Cerebral small vessel disease (SVD) is a vascular neurodegenerative disease which is the leading cause of vascular dementia and causes 20% of strokes. 20-30% of those over 80 show signs of the disease as white matter hyperintensities on MRI scans, doubling their risk of stroke and trebling their risk of dementia. Sporadic SVD is thought to be caused by hypertension but 30% of sufferers are normotensive and an alternative hypothesis implicates loss of integrity of the blood brain barrier (BBB). To investigate this, I studied brains from normotensive people with early stage SVD and found reduced capillary endothelial claudin-5 (a BBB tight junction protein), more oligodendrocyte precursor cells (OPCs; the precursors to myelinating oligodendrocytes), and more microglia/macrophages compared to controls. Furthermore, in a relevant rat model of spontaneous SVD, the Stroke Prone Spontaneously Hypertensive Rat (SHRSP; disease model; DM) I found that reduced endothelial claudin-5 was the earliest change, appearing at 3 weeks of age, followed by OPC proliferation, appearing at 4 weeks, and then increased number of microglia/macrophages, appearing at 5 weeks. Importantly, all these changes occurred at a young age (< 5 weeks), before any measurable hypertension. These changes were confirmed in an ex vivo slice culture model (i.e. removing blood flow), ruling out direct damage by leakage of blood components through an impaired BBB and suggesting an inherent endothelial cell dysfunction as the primary cause, with secondary BBB defects. This hypothesis of endothelial dysfunction is supported by increased endothelial cell proliferation in both human SVD tissue and the DM rats, and lower levels of endothelial nitric oxide synthase (eNOS) in brains of DM rats. To study this further I isolated primary brain microvascular endothelial cells (BMECs) from DM and control rats and found that those from DM rats formed less mature tight junctions (less membranous claudin-5) than control BMECs. I also found that conditioned media (CM) from DM BMECs causes OPCs in culture to proliferate more and mature less. This indicates that the endothelial dysfunction is inherent to the endothelial cells, rather than induced by other cell types, and through secreted factors causes OPC changes mirroring what is seen in vivo. Using an antibody array, I identified HSP90α as a candidate secreted factor and showed that it is necessary (by blocking the protein in CM) and sufficient (by adding recombinant HSP90α) to induce the maturation phenotype in OPCs, but not the proliferation phenotype. The idea that endothelial dysfunction causes SVD begs the question of what causes endothelial dysfunction, especially in our inbred DM rat strain. To establish this, I reanalysed sequencing data of the DM and control rats from a previously published study, searching for mutations which lead to truncated proteins in genes expressed in brain endothelial cells. We confirmed the candidate gene Atp11b, a phospholipid flippase, was mutated as predicted. I found that knocking down Atp11b using siRNA in a control endothelial cell line caused endothelial dysfunction and a loss of tight junction maturity, and that CM from these cells causes OPCs to proliferate more and mature less, mirroring what we see in primary DM BMECs and suggesting that Atp11b has a key function in promoting normal endothelial function. Furthermore, I showed that knocking down Atp11b causes cells to secrete increased levels of HSP90α. I propose a mechanism whereby ATP11B regulates the retention of HSP90α within endothelial cells, which in turns regulates eNOS levels and activity, as has been shown previously. In summary, this work shows that there are many pre-symptomatic changes which occur in the brain in the development of SVD in DM rats, and that these are ultimately caused by endothelial dysfunction. As these changes are similar to those found in spontaneous human SVD, I propose that endothelial dysfunction is a key mechanism of human SVD, which may in the future lead to new therapies.

A Ilha Comprida, localizada no litoral sul do Estado de São Paulo, é uma ilha-barreira arenosa holocênica de 63 km de comprimento por até 5 km de largura, limitada a SW pela desembocadura de Cananéia, e a NE, pela desembocadura de Icapara. Seu sistema praia - duna frontal teve grande variação no tempo, como evidenciado por sucessivos truncamentos de cordões litorâneos. Em escala de tempo mais restrita, dentro dos últimos 200 anos, coberta pelo registro histórico e cartográfico e por fotografias aéreas, as mudanças morfológicas mais significativas concentram-se no extremo nordeste da ilha. Incluem a variação de largura na área hoje caracterizada pelo máximo estrangulamento da ilha, a formação de um pequeno campo de dunas transgressivo nesta mesma área, o surgimento em 1943 de uma projeção da linha de costa, atualmente em estágio avançado de erosão, e o aparecimento intermitente e sucessivo de embaiamentos lagunares no setor adjacente da ilha de Iguape. A abertura do canal artificial do Valo Grande, em 1852, atuou de modo direto ou indireto em pelo menos parte destas mudanças. O aumento de vazão, de retrabalhamento de sedimentos arenosos e portanto de aporte de carga de fundo, devido ao Valo Grande, induziu o surgimento de ilhas arenosas e a formação de ?manchas de areia? no fundo do canal lagunar, bem como a intensificação da dinâmica de meandramento da desembocadura de Icapara, implicando possível aceleração na migração desta desembocadura rumo NE e no próprio crescimento longitudinal da barreira. O sistema praia - duna frontal da Ilha Comprida apresenta também grande variação no espaço, evidenciada pela alternância ao longo de sua extensão de morfologias diferentes, o que pode ser atribuído à intercalação entre setores de caráter deposicional e erosivo. As extremidades da barreira caracterizam-se pela ocorrência marcante de pequenos campos de nebkhas sobre terraços baixos de dunas frontais (<0,5 m). A presença dessas feições pode ser relacionada à contínua progradação próximo às desembocaduras lagunares de Icapara e de Cananéia. A zona erosiva mais evidente encontra-se na extensão centro-sul da praia, como evidenciado pela ocorrência de cordões antigos truncados pela linha de costa atual, e pela redução em tamanho de dunas frontais ativas incipientes e estabelecidas, as quais aparecem por vezes com falésias vivas associadas. Na mesma região, a morfodinâmica muda de regime dissipativo para intermediário, com presença de cúspides de espraiamento e berma. Esta região, ademais, concentra a mais alta variabilidade e complexidade de morfologia de dunas (i.e. cordões de dunas incipientes e estabelecidas obliquamente amalgamadas), além de uma concentração maior de dunas do tipo rampa. No restante do sistema, predomina ampla face de praia de baixa declividade (média de 0,66o), com tendência para formas mais estáveis de dunas frontais (terraços e cordões). Medidas estatísticas de distribuição granulométrica (diâmetro médio, desvio-padrão e assimetria) obtidas através do método dos momentos de Pearson, permitiram inferir, pela regra de McLaren, duas células de deriva litorânea longitudinal principais. A célula nordeste, dirigida para NE, abrange aproximadamente 53 km, e a célula sudoeste toma em sentido oposto (SW) os restantes 10 km. As duas zonas de pólo deposicional de deriva são caracterizadas por amplos e baixos terraços de dunas frontais incipientes progradacionais. A zona de divergência entre as células de deriva coincide com a região de erosão costeira mais intensa. A distribuição em massa de minerais pesados é concordante com o transporte longitudinal verificado. Responde a isso a tendência de aumento de minerais de menor equivalente hidráulico (em geral, quimicamente mais instáveis) a despeito da redução de minerais de equivalente hidráulico mais alto (em geral, quimicamente mais estáveis) no rumo de transporte. A integração dos resultados obtidos por análise morfológica (fotointerpretação e estudo de feições de campo) e sedimentológica (granulometria e minerais pesados) permitiu estabelecer um modelo morfodinâmico para o sistema praia - duna frontal recente da Ilha Comprida. Neste modelo, a fase final de evolução da ilha é atribuída tanto à superação do obstáculo ao crescimento longitudinal exercido pelo morro de Icapara, quanto ao incremento de carga sedimentar na desembocadura de Icapara, induzido pela abertura do Valo Grande. / Ilha Comprida is a Holocene sandy barrier-island located on the south coast of São Paulo State, 63 km long and 2 to 5 km wide. Its transverse boundaries are Cananéia and Icapara inlets, at the SW and NE, respectively. The beach-foredune system of Ilha Comprida have experienced intense morphological variation in time, as showed by the successive truncations of littoral ridges. In a more restricted time-scale (at least 200 years) as covered by historical and cartographic records and aerial photographs, the most relevant changes have occurred on the northeast extremity of the island. These changes include: i) variation in width at the present most narrow region; ii) formation of a little transgressive dunefield (2 km2) in the same region; iii) initiation (in 1943) of a projection in the coastline, actually in advanced erosional state; iv) intermittent and successive appearing of lagoonal embayments at the neighboring sector of the Iguape island. The opening of the Valo Grande artificial channel, in 1852, acted either directly and indirectly on these changes. The increase of sedimentary bed-load supply induced the appearance of sand islands and submerged sand patches along the lagoonal channel. Beside this, the increasing meandering of Icapara inlet forced its rapid migration to the NE, and the longitudinal growing of the barrier. The beach-foredune system also shows great variability alongshore, observed on the morphologic alternance of depositional and erosional sectors. The tips of the barrier are characterized by the marked occurrence of small nebkha fields on foredune terraces (up to 0.5 m high). These aeolian morphological features can be related with a continuous progradation nearly Icapara and Cananéia inlets. The most-evidenced erosional zone stays at the mid-southwestern portion of the beach, as observed on the truncation of former beach and foredune ridges by the present coastline, and on the reduction in size of active incipient and established foredunes (that often appears with active cliffs). In the same region, the coastal morphodynamics changes from dissipative to intermediary beach, with presence of swash cusps and berm. Moreover, this site concentrates the highest variability and complexity of dune morphology (e.g. obliquely amalgamated incipient and established foredune ridges), beyond a increasing occurrence of ramp foredunes. In the remains of the beach-dune system, large shorefaces of low declivity (0.66o in mean), with tendency to more stable shapes of foredunes (terraces and ridges), are dominant. Statistical measures on the distribuition of granulometric frequencies (mean diameter, standard deviation, skewness), calculated through the Pearson?s moments method, indicate according to the McLaren rule two main longshore drift cells. The northeastern cell, oriented towards the NE, comprises about 53 km. The southwestern cell, comprising the remaining 10 km, is oriented towards the SW. The two depositional poles of longshore drift cells are characterized by wide and flat progradational, incipient foredune terraces. The divergence zone among these longshore cells drift coincides with the zone of the most intense coastal erosion. The mass distribution of heavy minerals is in agree with the deduced directions of longshore transport. This fact is confirmed by the tendency of increasing concentration of minerals with minor hydraulic equivalence (usually, more unstable minerals) in despite of reduction of minerals with high hydraulic equivalence (usually, more stable minerals) along the net longshore drift direction. The integration of data obtained by morphologic analysis (interpretation of aerial photographs and field features) and sedimentology (grain size, heavy minerals), allowed to postulate a morphodynamic model for the recent beach-foredune system of Ilha Comprida. According to this model, the late phase of barrier evolution is related both to the surpassing an obstacle to longitudinal growth (Icapara hill), and to the increasing sedimentary input at the Icapara inlet, favored by the opening of the Valo Grande channel.

Sepse e suas complicações (sepse grave e choque séptico) ainda são a principal causa de morte nas unidades de terapia intensiva em todo o mundo. Estudos clínicos e experimentais têm demonstrado que na fase inicial da sepse a concentração plasmática de arginina vasopressina (AVP) está elevada. No entanto, durante o processo fisiopatológico os níveis plasmáticos da mesma permanecem inadequadamente baixos, apesar de haver hipotensão persistente. Uma das hipóteses sugeridas para essa deficiência relativa de AVP é a apoptose de neurônios vasopressinérgicos. Nosso objetivo foi identificar elementos envolvidos na morte celular hipotalâmica, além de avaliar o comportamento de células gliais e da barreira hematoencefálica (BHE) durante a sepse. Ratos Wistar foram submetidos à sepse por ligadura e punção cecal (CLP) ou não manipulados (naive) como controle e então divididos em dois grupos. No primeiro, foram perfundidos e os cérebros coletados para imunohistoquímica. Outro grupo foi decapitado para a retirada de sangue para dosagem de interferon- gama (IFN-?) e encéfalo para análise da expressão de proteínas no hipotálamo ou nos núcleos supraópticos (SON) e paraventriculares (PVN). Um terceiro foi separado para investigação da permeabilidade da BHE. Apesar de aumento da imunomarcação de CD8 e MHC-I no SON dos animais sépticos, não encontramos indícios de morte celular mediada por células imunes. No SON e PVN de animais sépticos, a expressão de fatores envolvidos na ativação da via extrínseca de apoptose (tBID, caspase-8 clivada) se manteve inalterada, enquanto fatores anti-apoptóticos relacionados à via intrínseca (BCL-2, BCL-xL) estavam diminuídos no hipotálamo. No SON destes animais a micróglia assumiu uma morfologia associada à sua ativação, concomitante com o aumento plasmático de IFN-?. Houve rompimento transitório da BHE no hipotálamo após 6 horas do CLP. Os resultados indicam que a via intrínseca de apoptose parece ser a responsável pela morte celular que é observada nos núcleos vasopressinérgicos e essa condição está temporalmente associada à ativação microglial e rompimento da BHE / Sepsis and its complications (severe sepsis and septic shock) remain as the main cause of death in intensive care units worldwide. Clinical and experimental studies have shown that in the early phase of sepsis the plasma concentration of arginine vasopressin (AVP) is increased. However, during the pathophysiological process the plasma levels remain inadequately low, despite of persistent hypotension. One of the hypothesis suggested for this relative deficiency is the apoptosis of vasopressinergic neurons. Our objective was to identify elements involved in the hypothalamic cellular death and evaluate the modifications of glial cells and blood-brain-barrier (BBB) during sepsis. Wistar rats were submitted to sepsis by cecal ligation and puncture (CLP) or non-manipulated (naïve), as control and then divided in two groups. In the first one, they were perfused and brains were collected for immunohistochemistry. In another one they were decapitated for blood collection and further plasma interferongama (IFN-?) analysis by ELISA. Brain was also collected for apoptosis-related proteins expression analysis in the hypothalamus or in the supraoptic (SON) and paraventricular (PVN) nuclei. A third set was separated for the investigation of BBB permeability. Despite of increased immunostaining for CD8 and MHC-I in the SON of septic animals, we did not find evidence of cell death mediated by immune cells. In the SON and PVN of septic animals, the expression of proteins involved in the activation of the extrinsic apoptosis pathway (tBID, cleaved caspase-8) was not altered, whereas anti-apoptotic factors related to the intrinsic pathway (BCL-2, BCLxL) were decreased. In the SON of these animals, microglia assumed a morphology related to its activation, associated with the increase of plasma IFN-?. There was a transitory breakdown of BBB in hypothalamus after 6 hours following CLP. The results indicate that the intrinsic apoptosis pathway seems to be responsible for the cell death observed in vasopressinergic nuclei and this condition is temporally associated with microglial activation and BBB leaking

Esta tese realizou-se com o intuito de melhor conhecer a proteção tarifária agrícola da União Européia (UE) e sua eventual associação com a Política Agrícola Comum (PAC). A hipótese implícita nesse trabalho é a de que há uma discriminação tarifária na pauta agrícola da UE; discriminação essa que estaria associada à estrutura da PAC. Nesses termos, o trabalho teve três objetivos complementares. Em primeiro lugar, buscou-se medir a magnitude das tarifas praticadas pela União Européia sobre os produtos agrícolas exportados pelo Brasil. Como segundo ponto, procurou-se identificar a existência de discriminação tarifária entre os grupos de produtos agrícolas presentes na pauta de tarifas do bloco europeu. E, por fim, avaliou-se a eventual existência de simultaneidade entre a proteção tarifária e as políticas domésticas de suporte à agricultura comunitária. A teoria visitada e os trabalhos já realizados com tal pano de fundo sugeriram que há grupos de produtos particularmente protegidos pelas tarifas agrícolas do bloco europeu. Esses mesmos trabalhos também apontaram para o caráter de permanência das políticas de apoio à agricultura da União Européia, mesmo após as reformas da PAC em 1992 e 2000. Tais políticas não são homogêneas; antes disso, estão concentradas em determinadas atividades e perfis de produções da agricultura do bloco. A revisão teórica também foi empregada para apresentar os principais instrumentos de política comercial da União Européia em itens agrícolas e para discutir os conceitos inescapáveis em barreiras tarifárias: tarifas ad valorem, tarifas específicas, mecanismo de gatilhos, picos tarifários, e quotas tarifárias. Cada um desses conceitos chave em política comercial foi avaliado com base em ferramentas estatísticas específicas, procurando-se identificar os produtos eventualmente discriminados em cada caso. Em paralelo, empregaram-se regressões lineares contra o tempo a fim de avaliar se tem ocorrido queda no nível de suporte à agricultura comunitária, bem como se procedeu à identificação dos setores menos contemplados pela oferta agrícola feita pela União Européia nas negociações do fórum biregional UE-Mercosul. Como resultado do esforço empreendido, esse texto pretende deixar duas contribuições. Primeiramente, através de simulações acerca do uso de diferentes preços de referência para a conversão das tarifas específicas e mistas em tarifas equivalentes ad valorem, no contexto da pauta agrícola comunitária. A discussão acerca dos impactos do uso de distintos preços de referência em situações similares pretende ser uma das contribuições do trabalho. Em segundo lugar, propõem-se indicadores especificamente projetados para a avaliação de duas questões importantes em acesso a mercado de bens agrícolas nos países desenvolvidos. O primeiro deles destinado à leitura da proteção concedida pelo mecanismo de quotas tarifárias, e o outro voltado à identificação de picos tarifários entre grupos de produtos. As conclusões do trabalho apontam para determinados grupos de produtos agrícolas alvo de proteção da parte dos instrumentos de política comercial na União Européia, destacando-se: carnes e miudezas, leite e laticínios, cereais, preparações de carne e peixes, açúcares e confeitaria, preparações de cereais, e tabaco e seus manufaturados. Esses grupos de produtos mostraram-se muito similares às produções agrícolas mais intensamente apoiadas pelas políticas de financiamento da PAC. / This thesis has been done to better understand the European Union (EU) agricultural tariff protection and its association with the Common Agricultural Policy (CAP). The hypothesis is that there is tariff discrimination in the EU agricultural tariff schedule and that such discrimination is connected with the CAP structure. For this purpose, the study is oriented by three goals, which complement themselves. First, it aims to measure the magnitude of the tariff used by the European Union on the agricultural products exported from Brazil. Second, it tries to identify the existence of tariff discrimination among the groups of agricultural products in the European tariff set. At last, it evaluates the existence of simultaneity between tariff protection and agricultural support policies in the EU. Theoretical and empirical studies have already suggested that there are groups of products particularly protected by the EU agricultural tariffs. These studies also pointed to the permanent character of the agricultural support policies in the EU, even after the CAP reforms in 1992 and 2000. These policies are not homogeneous; but instead, they are shaped according to certain activities and production profiles of the EU agricultural sector. The literature has also been used to present the main instruments of the EU trade policy on agricultural goods and to discuss the key concepts on tariff barriers: ad valorem tariffs, specific tariffs, trigger tools, tariff peaks, and tariff-rate quotas. With the purpose of identifying the products discriminated, each of the topics on trade policy is evaluated through the application of specific statistical tools. In order to evaluate if there has been a decrease in the level of agricultural support in the European Union we use linear time series regressions. At the same time, the sectors discriminated by the EU agricultural offer in the European Union-Mercosur negotiations are also identified. The collected data are compared to evaluate the agricultural products that are protected by the EU. The present work has two main contributions. Firstly, the discussion related to the effects of using different prices to the conversion of EU specific and composed tariffs into equivalent ad valorem tariffs. Secondly, tools specially designed to analyze two important questions on agricultural market access in developed countries are proposed. The first one focuses on capturing the protection given by tariff-rate quotas, and the other focuses on the identification of tariff peaks among sets of products. The conclusions of this study point to certain sets of agricultural products targeted by the EU trade policy, especially meat, dairy products, cereals, meat preparations, sugar, cereal preparations, and tobacco. These sets of products appeared to be quite similar to the agricultural products severely supported by CAP financing policies.

Dans le contexte énergétique actuel, la thermoélectricité, basée sur la conversion directe de l'énergie thermique en énergie électrique, est en plein essor. Cette technologie, qui permet de récupérer l'énergie perdue sous forme de chaleur au cours de processus industriels, est basée sur l'utilisation de modules thermoélectriques. Ces modules sont constitués de la juxtaposition de branches thermoélectriques, constituées de semi-conducteurs n et p, reliées par des jonctions électriques. Dans la gamme de températures de 300 à 600°C, les matériaux de type skutterudite RM4X12 (R : terre rare, M : métal de transition, X : pnictogène) présentent des propriétés thermoélectriques intéressantes. Cependant, les propriétés aux interfaces entre les jonctions électriques et le matériau thermoélectrique jouent un rôle très important dans la performance des modules thermoélectriques. Il est nécessaire de minimiser les pertes électriques et thermiques au niveau de ces interfaces. De même, en zone chaude principalement, il est impératif de limiter l'inter-diffusion entre les éléments constituant le matériau thermoélectrique et le matériau utilisé pour les connexions électriques. La formation de composés aux interfaces peut, en effet, être à l'origine de la dégradation des propriétés du matériau thermoélectrique. Ces contraintes conduisent à introduire des barrières de diffusion entre le matériau thermoélectrique et les connexions électriques. C'est dans ce contexte que ce situe la présente étude, l'objectif étant d'étudier la potentialité de matériaux à base de nitrure de tantale en tant que barrières de diffusion. Ainsi, des couches minces à base de nitrure de tantale ont été déposées, par le procédé de pulvérisation cathodique, sur les substrats de type skutterudite, CeFe4Sb12, les connexions électriques étant en cuivre. L'efficacité de barrières monocouches, TaN, et tri-couches, TaN/Ta/TaN, a été étudiée, ces couches présentant une épaisseur totale de 1 ou 1,5 µm. La première étape de ce travail a consisté en l'étude de la stabilité thermique du matériau skutterudite afin de déterminer le domaine d'utilisation en température de ce matériau. Dans une seconde étape, la potentialité de monocouches, TaN, et multicouches, TaN/Ta/TaN, en tant que barrières de diffusion a été déterminée à partir d'une étude microstructurale. Les assemblages CeFe4Sb12/barrière/Cu ont été préalablement soumis à des traitements thermiques sous vide à des températures variant de 400 à 600°C. Enfin dans une dernière étape, l'étude théorique des mécanismes de migration a été menée à partir des calculs d'énergies d'incorporation et de migration des atomes étrangers, tels que le cuivre et l'antimoine, dans le nitrure de tantale massif. / Due to the current energy context, thermoelectricity based on the direct conversion of thermal energy into electrical energy is of great interest. Direct conversion of thermal energy to electrical energy requires the use of thermoelectric devices made of n- and p-type semiconductor couples connected by electrical junctions. RM4X12 (R: rare earth, M: transition metal, X: pnictogen) skutterudite compounds have been reported to be promising for thermoelectric applications in the [400-600]°C intermediate temperature range. However the performance of thermoelectric devices is strongly dependent on the joining of thermoelectric couples with metal electrodes as the conversion efficiency is greatly influenced by the contact resistance. High electrical and thermal conductivities are required associated with a high interfacial mechanical strength. Moreover the joining material has to be selected to avoid any interfacial reaction occurring during the device fabrication and use. Diffusion barriers are also needed to limit these interfacial reactions which may be detrimental to the thermoelectric device performance. The aim of this work is to study the efficiency of tantalum nitride based materials as diffusion barriers. TaN single layer and TaN/Ta/TaN multilayers barriers were deposited by sputtering between the CeFe4Sb12 skutterudite substrate and the Cu electrical junction. The inter-diffusion of elements was studied through these mono- and multi-layers, of 1 μm or 1.5 μm in thickness. In a first step, the thermal stability of the skutterudite has been investigated to determine the use temperature range of this material. In a second step the efficiency of TaN and TaN/Ta/TaN layers as diffusion barriers has been determined from a microstructural study. CeFe4Sb12/barrier/Cu stackings were previously annealed under vacuum in the [400-600]°C temperature range. Finally a theoretical study of the migration mechanisms was carried out from the calculations of the incorporation and migration energies of species, more specifically Sb and Cu, in the bulk tantalum nitride.

Bakgrund: Postpartumdepression drabbar mellan 8-15 procent av kvinnor som föder barn. Depressionen bryter oftast ut några veckor efter förlossningen och kan vara upp till ett år eller mer och innebär ett stort lidande för den som drabbas. Studier visar att en stor del av de som drabbas inte söker hjälp, vilket visar på behovet av att sjuksköterskan fångar upp dessa kvinnor för att tidigt erbjuda stöd och behandling. Syfte: Syftet med litteraturöversikten var att undersöka vilka barriärer kvinnor drabbade av postpartumdepression upplever för att söka hjälp. Metodbeskrivning: Allmän litteraturöversikt valdes som metod, tio kvalitativa studier valdes utifrån databaserna PubMed och CINAHL. Artiklarna kvalitetgranskades utifrån en kvalitetsmall. Tre kategorier utformades därefter av den analyserade datan. Resultat: De kategorier som identifierades som barriärer var; kvinnorna själva, familj och omgivning samt sjukvården. Det identifierades en stor okunskap om postpartumdepression bland de deltagande kvinnorna, vilket ledde till att de normaliserade sina symtom och avvaktade med att söka hjälp. Rädslan för att bli stigmatiserade och att uppfattas som dåliga mammor var ytterligare en barriär. Bristande stöd från sjukvården framkom som ytterligare barriärer. Slutsats: Kvinnor upplever ett stigma kopplat till psykisk ohälsa och moderskap, vilket leder till att de inte söker hjälp. En ofta upplevd oförståelse inför sina symtom leder till att kvinnor normaliserar dem. En stöttande sjukvård är av största vikt, sjuksköterskans arbete kan bidra till att bryta stigmat och skapa en trygg och fördomsfri miljö där kvinnor kan öppna upp sig. / Background: Postpartum depression affects around 8-15 percent of women who have given birth. Symptoms present within a few weeks and can last upwards of a year; and cause a great deal of suffering to the women experiencing it. Evidence suggests women do not seek help for their depressive symptoms, which highlights the need of nurses to identify symptoms and offer treatment and support. Aim: The aim of this literature review is to examine the experienced barriers among women to seek help for postpartum depression. Method: The studies analysed in this literature review were chosen from the databases PubMed and CINAHL. Ten studies were included. Quality of the articles was identified using a criteria and three categories were found within the findings. Results: The three categories concerned the women, their surrounding and the health care as barriers. Stigmatisations attached to mental healt issues and the ‘ideal’ image of motherhood created a major barrier to seek help. Women experienced a lack of knowledge and understanding of PPD, they often normalised the symptoms and explained them with external factors. A lack of support from health care also posed as barriers when seeking help. Conclusion: Women experience numerous stigmatisations related to mental health issues and the ‘ideal’ image of motherhood, which figured as a major barrier to seek help. This study reiterates the importance of having supportive health care; with the nurse playing an important role in breaking this stigma and creating a safe environment for women to share their experiences.

Devant l'impact des maladies neurodégénératives sur la société, les thérapies par ultrasons focalisés apparaissent comme des techniques prometteuses combinant non invasivité, précision spatiale millimétrique et capacité d'atteindre les structures profondes du cerveau. Cependant, des travaux sont encore nécessaires pour renforcer les effets de la neuromodulation, comprendre les mécanismes sous-jacents et contrôler la sûreté de la technique avant d'entreprendre des essais cliniques. Dans cette thèse, la propagation des ultrasons dans le cerveau de rongeurs et de singes a été étudiée numériquement afin d'estimer l'intensité acoustique dans le cerveau, la répartition spatiale des ondes dans la boîte crânienne et l'élévation de température. Afin d'évaluer physiologiquement les effets des ultrasons à l'échelle cellulaire, l'activité de neurones uniques a été mesurée sur des macaques éveillés pendant une neuromodulation ultrasonore. Puis, la durée de l’effet de modulation a été augmentée grâce à une prolongation du tir sur des singes exécutant une tâche visuelle. L'imagerie fonctionnelle par IRM a ensuite permis de faire ressortir des changements de connectivité entre l'aire stimulée et des régions du cerveau éloignées.Enfin, les avantages de la neurostimulation par ultrasons ont été combinés avec l'efficacité d'un agent neuroactif. En utilisant des microbulles conjointement aux ultrasons, la barrière hémato-encéphalique a été ouverte localement et réversiblement dans le cortex visuel de macaques anesthésiés pour permettre le passage d'un neurotransmetteur inhibiteur dans le cerveau. La baisse d'amplitude des réponses EEG du cortex visuel à des stimuli démontre la faisabilité de la délivrance locale et non invasive de neuromodulateurs dans le cerveau. Ainsi, les paramètres ultrasonores ont été optimisés grâce aux simulations numériques et à des expériences in vivo pour renforcer les effets de neuromodulation tout en contrôlant les effets indésirables, avec l'objectif de se diriger vers des applications thérapeutiques et proposer de nouveaux outils pour des études de connectivité cérébrale / Considering the extent of neurodegenerative diseases consequences on the society, focused ultrasound appears as a promising technique combining non-invasiveness, millimetric spatial accuracy and ability to reach deep brain structures. However, efforts still need to be made to amplify the effects of focused ultrasound neuromodulation, understand its mechanism and control the safety of the technique before moving towards human trials.The ultrasound propagation inside the brain of rodents and monkeys was first studied numerically to estimate the maximum intensity in the brain, the pressure distribution in the skull cavity and the thermal rise. To evaluate physiologically the ultrasound effects at the cellular level, the activity of individual neurons was measured on awake macaques during ultrasonic neuromodulation. To further increase the duration of the modulation, a longer sonication was tested successfully on macaques performing a visual task. Functional MRI was then used to highlight the connectivity changes between the stimulated area and distant cerebral regions. Finally, the advantages of ultrasound neurostimulation were combined with the efficiency of a neuroactive agent (GABA). Using microbubbles and ultrasound, the blood brain barrier was opened locally and reversibly in the visual cortex of anesthetized macaques to deliver an inhibitory neurotransmitter in the brain. The amplitude of the EEG response of the visual cortex to stimuli decreased after GABA injection, demonstrating the feasibility of delivering neuroactive drugs non-invasively and locally to any brain region.Overall, ultrasound parameters were optimized with both numerical tools and in vivo experiments to amplify neuromodulation effects while controlling the safety. This work opens the way to the development of novel therapeutic applications and new tools for connectivity studies