Made available in DSpace on 2016-06-02T20:16:43Z (GMT). No. of bitstreams: 1 1994.pdf: 1761798 bytes, checksum: 183ce1c471cf1079a1302b309c923a68 (MD5) Previous issue date: 2002-04-11 / Financiadora de Estudos e Projetos / In the present work the Drude weight is calculated to study the capacity of conduction at finite temperatures in one-dimensional rings with the potencial given by the sequences of different degrees of randomness and threaded by a time independent magnetic flux. In this context, the tight-binding approximation is used to solve the Schrödinger equation,also permitting the potencials, given by substitution sequences, to be implemented in a simple way. We study four situations in which the potencial is given by periodic and random sequence. The transport properties exhibited by the system in each case are to reflect the degree of randomness presented by these potencials, although in a nontrivial manner if one considers a hierarchy of disorder based on the Fourier transform of the sequences. The role played by temperature in the capacity of conduction, as compared with the role played by the potencials, is also described. / No presente trabalho, através do cálculo do peso de Drude estudamos a capacidade de condução a temperaturas finitas, em anéis unidimensionais com potenciais dados por seqüências de diferentes graus de desordem e atravessados por fluxo magnético independente do tempo. Neste contexto, utilizamos o modelo tight-binding na resolução da equação de Schrödinger, o que permite inserir os potenciais, dados por seqüências de substituição, de maneira simples. Estudamos quatro casos em que esse potencial seja dado por seqüências não periódicas. Os resultados são comparados com os dois casos limites de organização, a saber, as seqüências periódica e desordenada. Observa-se que o grau de desordem apresentado por estes potenciais reflete-se nas propriedades de transporte exibidas pelo sistema em cada caso, embora de maneira não trivial se considerada uma hierarquia de desordem baseada na transformada de Fourier das seqüências. Descreve-se também a influência da temperatura, frente à influência dos potenciais, na capacidade de condução.

A assistência aos pacientes com doenças neurológicas de longa duração não é reconhecida na produção científica brasileira; entende-se por doença neurológica de longa duração: doenças neurológicas de curso longo, fase final de difícil identificação, com sintomas variados e condição de dependência moderada ou grave e comprometimento cognitivo de difícil avaliação. Neste estudo, foram consideradas as seguintes condições: estado vegetativo persistente, estado mínimo de consciência e síndrome do encarceramento (ou síndrome de locked-in). Em busca bibliográfica, verificou-se que não há artigo em língua portuguesa sobre o tema. No cotidiano, a assistência é focada em higiene e observação clínica. Entretanto, na perspectiva do Cuidado, algo mais é necessário ao se pensar este tipo de prática, para a promoção da dignidade humana. Os objetivos deste estudo são: relatar uma prática junto a pessoas com doenças neurológicas de longa duração focada no seu resgate biográfico, e fundamentar a mesma de maneira interdisciplinar, a partir do Cuidado como categoria reconstrutiva. É uma pesquisa qualitativa, na qual foram utilizados primariamente registros de um diário de campo da primeira autora, sobre intervenções em Terapia Ocupacional com esta população; a partir destes, foram produzidas narrativas e reflexões sobre a metodologia de intervenção. A partir do resgate biográfico, busca-se resgatar a condição de sujeito do paciente neurológico muito grave. Na segunda etapa, tem-se a fundamentação teórica numa perspectiva interdisciplinar; o conceito de Cuidado é a referência principal, compreendido como o sentido existencial da experiência de adoecimento. Também foi utilizado o referencial da psicanálise de orientação freudiana e lacaniana para aprofundamento teórico dos achados empíricos descritos através de narrativas. A reconstrução da biografia do paciente ocorre por diversas perspectivas, mas especialmente pela estimulação sensorial - não com foco de recuperação neurológicas, mas a partir do repertório do paciente e em busca da validação de sua condição de sujeito. Junto com uma detalhada avaliação e acompanhamento, o terapeuta faz um monitoramento contínuo, propondo atividades e dando especial atenção ás reações observadas, para que possa nomeá-las ou traduzi-las, bem como discriminar os estímulos agradáveis e desagradáveis; isto também possibilita ampliação das possibilidades de encontro deste sujeito com o mundo. Os conceitos psicanalíticos: estágio do espelho, real, lalíngua e constituição do sujeito apresentam-se como fundamentais para a compreensão das experiências empíricas vividas e, posteriormente, elaboração de uma fundamentação teórica. Memória, história, identidade, dignidade e subjetividade qualificam a prática junto a pessoas com doenças neurológicas de longa duração. A despeito das reações neurológicas apresentadas, tal prática possibilita a reconstrução de um cotidiano significativo e as teorias psicanalíticas de orientação freudiana e lacaniana constituem importante marco para fundamentação da reconstrução da assistência / The care of people with Long Term Neurological Conditions (LTNC) isn´t recognized in the Brazilian Palliative Care scientific production; in daily basis, the assistance is focussed on hygiene´s care and clinical observation. However, from the perspective of palliative care, something more is needed, to promote the dignity of the human person. To describe an occupational therapy intervention for patients with LTNC based on biographical rescue. Records and field diaries of an Occupational Therapy intervention promoted by the first author were used to produce narratives and reflections about this intervention´s methodology. The concept of Comprehensive Care is the main reference, designed as care interested in the existential sense of the illness process. It was also used the framework of Freud´s and Lacan´s psychoanalysis for theoretical studies of the empirical findings reported through narratives. The beginning of the reconstruction of the patient´s biography occurs by several perspectives, especially through sensory stimuli related to prior patient´s occupational repertoire. Along with the detailed evaluation, the therapist does a careful and continuous monitoring, proposing activities, giving special attention to their reactions, being able to discriminate what pleases or not, and expanding the possibilities of encounter of people with LTNC and the environment which surround them. The psychoanalytic concepts: mirror stage, real, lalangue and subject´s constitution had major importance to understand the empirical experiences and to develop a theoretical basis. Memory, history, identity, dignity, and subjectivity qualify the practice of occupational therapist with people with LTNC. Regardless of neurological reactions presented by the patients this practice helps reconstructing the meanings of the daily life of person with so severe disability. Sensory stimuli are shown as potential tools for a reconfiguration of the assistance with someone seriously affected by a neurological disease. Psychoanalytic theories of Freudian and Lacanian orientation are also an important framework for the reconstruction assistance

A luminescência persistente é um fenômeno em que o material emite radiação de segundos a várias horas após cessada a irradiação (luz, radiação UV, feixe de elétrons, etc.). No entanto, os mecanismos que geram o fenômeno da luminescência persistente ainda não são totalmente estabelecidos. Neste trabalho os materiais Lu2O3:TR3+,M (TR,M: Pr,HfIV; Eu, Ca2+ ou Tb,Ca2+) foram preparados pelo método de estado-sólido assistido por micro-ondas (MASS) e comparados com aqueles sintetizados pelo método cerâmico. As vantagens do método MASS incluem curto tempo de processamento, aquecimento dielétrico seletivo, baixo consumo de energia e uso de equipamentos de baixo custo (forno micro-ondas doméstico), muitas vezes produzindo produtos de alta pureza e alto rendimento. Os materiais foram caracterizados pelas técnicas de espectroscopia de absorção na região do infravermelho (IR), espectroscopia Raman, difração de raios X método do pó (DRX), microscopia eletrônica de varredura (MEV), X-ray absorption near edge structure (XANES), Extended X-ray absorption fine structure (EXAFS), X-ray Excited Optical Luminescence (XEOL), espectroscopia de fotoluminescência na região do UV-Visível, espectroscopia de fotoluminescência na região do UV-UV vácuo e termoluminescência (TL). Os fósforos Lu2O3:TR3+,M (TR,M: Pr,HfIV; Eu, Ca2+ ou Tb,Ca2+) foram preparados em um curto período de tempo (22-26 min) pelo método MASS utilizando forno micro-ondas doméstico, carvão ativado como susceptor, fluxos (H3BO3 ou Na2CO3) e sem a aplicação de gases. Todos materiais preparados com fluxo de H3BO3 exibem impurezas de LuBO3 que foram quantificadas por refinamento Rietveld. Os fluxos e os dopantes não alteraram consideravelmente a estrutura cristalina da matriz C-Lu2O3. As micrografias MEV sugerem que o fluxo de Na2CO3 e os precursores nitratos geram partículas de Lu2O3 com tamanho menor devido a evolução de gases provenientes da decomposição destes compostos. Por outro lado, quando é usado óxidos como precursores os materiais apresentam maiores tamanhos de partícula e na presença de H3BO3 leva a maior agregação. Os dados de XANES indicam que houve completa redução do íon TbIV &#8594 Tb3+ e parcial do PrIV &#8594 Pr3+, devido ao uso de carvão ativado que gera CO(g) durante o tratamento térmico. Os espectros da luminescência persistente indicam emissões nas regiões do vermelho/NIR, vermelho alaranjado e verde atribuídas as transições 4fN &#8594 4fN características dos íons Pr3+, Eu3+ e Tb3+, respectivamente. As diferenças entre os espectros registrados sob excitação UV e após cessada a irradiação podem ser explicadas pela emissão da luminescência persistente predominante dos íons TR3+ no sítio S6 do que no C2. Além disso, a co-dopagem aliovalente com os íons HfIV e Ca2+ aumentam a intensidade e duração da luminescência persistente. Isto ocorre através da geração de armadilhas provenientes dos dois co-dopantes nos sítios de Lu3+ e por defeitos produzidos na compensação de carga. Os materiais fotônicos preparados pelo método MASS com fluxo de H3BO3 apresentam maior intensidade e duração da luminescência persistente comparados aos preparados pelo método cerâmico ou sem a presença de H3BO3. Os mecanismos da luminescência persistente foram desenvolvidos através de princípios similares baseados nos dados experimentais da energia do band gap, posição dos níveis de energia dos íons TR3+/2+ na matriz e energia das armadilhas. Isto confirma a solidez da interpretação dos dados experimentais dos materiais Lu2O3:TR3+,M exibindo luminescência persistentes e encoraja a expansão de modelos similares para outros materiais apresentando esse fenômeno. Os fósforos Lu2O3:Pr3+,HfIV,Lu2O3:Eu3+(,Ca2+) e Lu2O3:Tb3+,Ca2+) apresentaram sintonização de cores de emissão tanto para o fenômeno da fotoluminescência como da luminescência persistente, podendo atuar como bons candidatos nas aplicações de bioimageamento ou sensibilizadores de células solares. / Persistent luminescence is a phenomenon where the material emits radiation from seconds to several hours after cessation of irradiation (light, UV radiation, electron beam, etc.). The persistent luminescence mechanisms are not entirely established, however. In this work, the materials Lu2O3:TR3+,M (TR,M: PrHfIV; Eu, Ca2+ or Tb,Ca2+) were prepared by MASS method as well as compared to these materials synthetized by ceramic method. The advantages of MASS method include short processing time, selective dielectric heating, low energy consumption and use of inexpensive equipment (domestic microwave oven), often affording high-purity and high-yield products. The materials were characterized by Infrared absorption spectroscopy (IR), Raman spectroscopy, X-ray powder diffraction (XPD), Scanning electron microscopy (SEM), X-ray absorption near edge structure (XANES), Extended X-ray absorption fine structure (EXAFS), X-ray excited optical luminescence (XEOL), photoluminescence spectroscopy in the UV-Visible range, photoluminescence spectroscopy in the UV-UV vacuum region and thermoluminescence (TL). The phosphorsLu2O3:TR3+,M (TR,M: Pr,HfIV; Eu, Ca2+ or Tb,Ca2+) were rapidly (22-26 min) and successfully prepared by MASS method using a domestic microwave oven, carbon as susceptor, fluxes (H3BO3 or Na2CO3) and without special gases application. All materials prepared with H3BO3 flux exhibit LuBO3 impurities that were quantified by Rietveld refinement. The flux and dopants does not considerably affect the crystalline structure of the C-Lu2O3 host matrix. Scanning electron micrographs suggest that Na2CO3 flux and nitrates precursors produce Lu2O3 particles of small size due to the gases evolution from the decomposition of these compounds. On the other hand, the materials prepared from oxides precursors have particles of large size and H3BO3 flux induces particle xi aggregation. The carbon used as the susceptor generates CO gas, leading to complete reduction of TbIV to Tb3+ and partial conversion of PrIV to Pr3+ present in the Tb4O7 and Pr6O11 precursors, as indicated by XANES. Persistent luminescence spectra of the materials show emission in the red/NIR, reddish orange and green ranges assigned to the 4fN &#8594 4fN transitions characteristics of Pr3+, Eu3+ and Tb3+ ions, respectively. Differences between the spectra recorded under UV excitation and after ceased the irradiation can be explained by the predominant persistent luminescence emission of TR3+ ion in the S6 site rather than TR3+ in the C2 site. In addition, inclusion of HfIV and Ca2+ codopants in the Lu2O3 host increases the emission intensity and duration of persistent luminescence due to generation of traps caused by charge compensation in the lattice as well as these metal ions in the Lu3+ sites. The photonic materials prepared by MASS method with H3BO3 flux show higher persistent luminescence performance than those prepared by the ceramic method or MASS without flux. The persistent luminescence mechanisms were developed through similar principles based on experimental data of band gap energy, energy level positions of TR3+/2+ ions in the host and traps energy. This similarity confirms the consistency of the interpretation of experimental data for the Lu2O3:TR3+,M materials and encourages the expansion of similar models for other persistent luminescence materials. Color tuning of persistent luminescence in Lu2O3:TR3+,M (TR,M: Pr,HfIV; Eu,Ca2+ or Tb,Ca2+) provides potential applications in bioimaging as well as in solar cell sensitizers.

Les méthodes de calcul en fatigue à grande durée de vie sont en cours de développement depuis des décennies et sont utilisées par les ingénieurs pour dimensionner les structures. Généralement, ces méthodes se basent sur la mise en équations de quantités mécaniques calculées à l'échelle macroscopique ou mésoscopique. Les critères de fatigue multiaxiale reposent généralement sur des hypothèses de changement d'échelle dont l'objectif est d'accéder à l'état de contraintes ou de déformations à l'échelle du grain. Dans les approches de type plan critique (Dang Van, Papadopoulos, Morel), l'amorçage d'une fissure de fatigue est considéré comme piloté par une quantité mécanique liée à une orientation matérielle particulière (plan critique). Si ces phénomènes sont bien établis dans le cas des chargements uniaxiaux, la nature des mécanismes liés à l'activation des systèmes de glissement, à la multiplicité du glissement et aux différents sites préférentiels d'amorçage de fissures sous chargements complexes reste peu connue.Afin de mieux comprendre les mécanismes d'endommagement en fatigue multiaxiale, les techniques d'analyse et de caractérisation de l'activité plastique (activation des systèmes de glissements, bandes de glissement persistantes) et d'observation de l'endommagement par fatigue ont été mises en place en se basant principalement sur des observations MEB et analyses EBSD. Ces investigations ont permis de mettre en lumière les effets des chargements non proportionnels sur la multiplicité du glissement sur du cuivre pur OFHC. L'étude statistique des sites préférentiels d'amorçage de fissures montre que les grains à glissement multiple présentent une forte probabilité d'amorçage de fissures, surtout sous les chargements non proportionnels. Nous avons également mis en évidence le rôle des joints de grains et des joints de macle sur le développement de la plasticité à l'échelle de la microstructure. Les résultats expérimentaux sont confrontés à ceux du calcul éléments finis (EF) en plasticité polycristalline sur des microstructures synthétiques 3D semi-périodiques. L'application du critère de Dang Van à l'échelle mésoscopique (le grain) montre une forte variabilité de la contrainte hydrostatique et du cisaillement. Cette variabilité est plus importante pour un modèle de comportement cristallin élastique anisotrope. Le rôle de la plasticité cristalline se révèle secondaire. Ces analyses permettent de remettre en perspective les hypothèses usuelles de changement d'échelle utilisées en fatigue multiaxiale. Enfin, une méthode basée sur la statistique des valeurs extrêmes est proposée pour le dépouillement des calculs EF sur agrégats. Cette analyse a été appliquée sur la contrainte équivalente associée au critère de fatigue de Dang Van pour les calculs d'agrégats polycristallins avec différentes morphologies et orientations des grains. Les effets de la surface libre, du type de chargement et du modèle de comportement mécanique des grains ont été analysés. Les résultats offrent des perspectives intéressantes sur la modélisation de l'amorçage des fissures en fatigue multiaxiale des matériaux et des structures avec une prise en compte de la microstructure. / The development of high cycle fatigue (HCF) strength assessment methods has now been running for more than a century, leading to relatively efficient methods for engineers. Generally, these methods are based on mechanical quantities calculated at macroscopic or mesoscopic scales and validated by the model's ability to accurately reproduce experimental results. Multiaxial fatigue strength criteria are usually based on scaling transition assumptions aiming at capturing the stress or strain state in the grain. In the case of critical plane based criteria (Dang Van, Papadopoulos, Morel), fatigue crack initiation is supposed to be controlled by a mechanical quantity linked to a particular orientation (critical plane). If fatigue crack initiation phenomena are well established in the case of uniaxial loadings, the nature of the mechanisms involved in the activation of slip systems, multiple slip and preferential sites of rack initiation under complex loadings remains little known.To better understand the mechanisms of multiaxial fatigue crack initiation, analysis and characterization of the plastic activity (e.g. activation of slip systems, persistent slip bands) and observations of fatigue damage have been carried out on pure OFHC copper, using SEM and EBSD analyses. These investigations enabled to highlight the effects of non-proportional multiaxial loadings through the induced multiplicity of slip. The statistical study of preferential crack initiation sites shows that grains with multiple slip have a high probability of crack initiation, especially under non-proportional loading. We also highlighted the role of grain boundaries and twin boundaries on the development of plasticity across the microstructure. The experimental results were compared with those of finite element crystal plasticity computations on synthetic 3D semi-periodic microstructures. The application of the Dang Van criterion at the mesoscopic (grain) scale showed a strong variability of the hydrostatic stress and the shear stress. This variability was greater for anisotropic elastic behavior, while the role of crystal plasticity seemed to be secondary. These analyses allowed putting into perspective the usual assumptions of scaling transition rules used in multiaxial fatigue. Finally, a method based on the extreme values statistics ​​was proposed and applied to the equivalent stress associated to the Dang Van fatigue criterion for polycrystalline aggregate computations with different morphologies and grains orientations. The effects of the microstructure, free surface, loading types and mechanical behavior were analyzed. The results offered interesting insights into the multiaxial fatigue modeling of metals and structures taking into account the microstructure.

Tee PhD thesis presents the study of the response of Mycobacterium tuberculosis, the causative agent of tuberculosis, upon prolonged exposure to lethal concentrations of the first line anti-tuberculosis drug, rifampicin. The study shows that prolonged exposure to lethal concentration of rifampicin causes cell death initially by several log orders of cells, followed by a persistence phase, from where rifampicin resisters emerge carrying mutation in the RRDR locus of the rpoB gene. This phenomenon was found to occur even when the drug concentration is well above MBC levels. Luria-Delbruck experiment, in a modified format, showed that the resisters emerged as fresh mutants, and not due to the growth of pre-existing natural resisters to rifampicin. The per sister cells, which showed high levels of hydroxyl radical generation, were found to have thickened outer layer, unlike the mid-log phase cells, which restricts the permeability of a fluorescent-conjugate of rifampicin, 5-FAM-rifampicin, 10-fold less in per sister cells, as compared to mid-log phase cells. The thickened outer layer has high negative surface charge and is hydrophilic in nature. It is proposed that the hydrophilic-natured thickened outer layer might have restricted the permeability of lethal concentrations of hydrophobic-natured rifampicin. This, in turn, might have ensured the presence of sub-lethal concentration rifampicin inside the per sisters, which in turn might have generated the hydroxyl radical that caused mutagenesis to generate rifampicin resisters. The Chapter 1 is the Introduction to the thesis presented in 4 parts – Part 1.1, 1.2, 1.3 and 1.4, introducing briefly the history of antibiotics, antibiotic resistance, antibiotic persistence and a brief history of tuberculosis respectively. It is concluded with a rationale behind the present study. The Chapter 2 presents the entire Materials and Methods used in the experiments described in the thesis. The Chapter 3 presents the data on the response of M. tuberculosis to rifampicin upon extended exposure. Rifampicin exposure of susceptible M. tuberculosis H37Ra cells showed a decrease in their CFU/ml followed by a persistence phase wherein the CFU/ml remained constant. However, prolonged exposure of rifampicin even at higher concentrations showed regrowth in the culture, which was found to be due to the emergence of rifampicin-resistant bacteria. Screening of rifampicin-resistant mutants showed point mutations in the rifampicin resistance determining region (RRDR) of the rpoB gene in all the mutants. In parallel, using trans formants of M. tuberculosis expressing unstable GFP under the respective native ribosomal RNA promoter, the metabolic status of the per sister cells was determined. When actively growing highly fluorescing cells were exposed to lethal concentration of rifampicin, their metabolism diminished, as illustrated by the decrease in their fluorescence during persistence phase, followed by the emergence of a sub-population of bacteria which were again metabolically active. In order to verify whether the rifampicin resisters are freshly formed mutants or have come from the naturally existing resisters, Luria-Delbruck fluctuation test was performed in a modified manner. The number of rifampicin resisters that emerged from the persistent phase was found to vary amongst different cultures from different days and different times of exposure, showing fluctuation. However, the addition of theorem before persistence phase almost abolished the generation of the rifampicin-resistant bacilli, indicating the role of hydroxyl radical in the emergence of rifampicin resisters. The generation of hydroxyl radical in mycobacteria exposed to rifampicin was confirmed using electron para-magnetic resonance spectrometry (EPR), with the spin trap agent, 5,5- Dimethyl-1-pyrroline N-oxide (DMPO) specific for hydroxyl radical. An increase in the formation DMPO-OH adduct in the persistence phase cells was observed, in comparison to mid-log phase cells. Exposure to theorem significantly diminished the adduct formation. The persistence phase cells also showed significantly high levels of signal specific to the hydroxyl-specific fluorescent dye, hydroxyphenyl fluorescein (HPF), as compared to the mid-log phase cells. In addition we have determined the oxidative stress in the bacilli upon rifampicin exposure using a redox biosensor (Mrx1-roGFP) which also showed high oxidative stress in the persistent phase. These observations confirmed the presence of high levels of oxidative stress and hydroxyl radical in the rifampicin persistent cells, in comparison with mid-log phase cells. Whole genome sequencing of the four independently isolated rifampicin resistant M. tuberculosis showed genome wide mutations having less common mutations with respect to the wild type genome, indicative of the occurrence of random mutagenesis. In addition mutation frequencies were comparable between the samples with respect to the wild type sample. About 69% of the mutations were A-C or T-G, followed by A-T or T-A, which is known to be due to oxidative stress in the cells. Variations in the colony morphology were also observed on the persistent phase cells, indicating the occurrence of mutagenesis in the bacterial genome during rifampicin treatment. The Chapter 4 is on the morphological and ultrastructural studies on rifampicin-exposed M. tuberculosis cells. Transmission electron micrographs of rifampicin per sisters showed significant thickening of the outermost capsular layer (OL), as compared to the mid-log phase cells. This observation was verified by staining the cells with ruthenium red, which specifically stains anionic polysaccharide of the OL. Zeta potential (ZP) measurement of the surface charge of the persistent cells showed high negative ZP, as compared to the mid-log phase cells. The negative ZP value was found to gradually increase during the course of rifampicin treatment and to decrease during the regrowth phase. Hexadecane assay showed larger proportion of per sister cells being retained in the aqueous phase, as compared to the mid-log phase cells. This indicated the higher hydrophilicity of the per sister cells, which was in agreement with the higher surface negative charge of the cells. The permeability of rifampicin per sister cells to 5-FAM-rifampicin (rifampicin conjugated to 5-carboxy fluorescein, which is as hydrophobic as rifampicin) was found to be 10-fold less than that of the mid-log phase cells. However, removal of the thick OL by bead beating (BB) with 4 mm glass beads significantly improved the permeability of per sister cells to 5-FAM-rifampicin. On the contrary, no difference in the 5-FAM-rifampicin uptake was observed in mid-log phase cells, with or without BB. These observations implied that the thick hydrophilic-natured OL with high negative surface charge may be playing a significant role in limiting the permeability of hydrophobic-natured rifampicin entry into the persisted cells. This in turn may ensure the presence of sub-lethal concentration of rifampicin inside the persisted cells that are exposed to lethal concentration of the antibiotic. Exposure of bacteria to sub-lethal concentration of antibiotics has been reported to generate oxidative stress in the bacteria, leading to mutagenesis. A model has been proposed based on these observations in which the persistent mycobacteria are protected from lethal concentrations of the rifampicin by the thick OL which in turn ensures sub-lethal intracellular antibiotic concentration, leading to the generation of hydroxyl radical mediated mutagenesis and thereby emergence of rifampicin resisters. This thesis is concluded with the list of salient findings, publications and references.

INTRODUCTION. Congenital diaphragmatic hernia (CDH) combines a congenital malformation of the diaphragm with lung hypoplasia, leading to severe respiratory distress and intractable pulmonary hypertension of the newborn. Despite advances in prenatal diagnosis and neonatal intensive care, CDH is associated with high mortality and devastating morbidities. In the absence of curative treatment, numerous prenatal therapies have been used experimentally with varying success. So far, only fetal tracheal occlusion has been tested in clinical trials, but the consequences for the human lung are poorly known. AIMS. To further characterize the rabbit model of CDH, which was subsequently used to assess the effects of prenatal therapies on airway and pulmonary vascular development, including tracheal occlusion, and two novel approaches, perfluorooctylbromide and an activator of soluble guanylate cyclase (BAY 41–2272), which were given through tracheal instillation.METHODS. After a diaphragmatic incision during the pseudoglandular stage, fetal rabbits were randomized against placebo/sham operation during the saccular stage for tracheal occlusion, perfluorocarbon or BAY 41–2272. At term operated fetuses and controls were subject to evaluation of lung mechanics and/or hemodynamics as well as postmortem lung analyses. Human fetal and neonatal lung tissue, including controls and CDH with tracheal occlusion or expectant management, was analyzed histologically and biochemically.RESULTS. The rabbit model of CDH was characterized by reduced lung volumes and impaired compliance, disorders of elastin deposition within alveolar walls, and downregulation of elastogenesis-related genes. Moreover, this model reproduced features of pulmonary hypertension, including high right ventricular pressure and level of N-terminal-pro-B type natriuretic peptide, remodeling of pulmonary arterioles, decreased alveolar capillary density, and downregulation of vasodilation-related genes. In the rabbit model, lung distension caused by tracheal occlusion improved alveolar formation and elastogenesis, yet without correction of lung mechanical parameters. Tracheal occlusion increased also the expression of other extracellular matrix components, which reflected myofibroblast activity, and reduced the transcription of surfactant-associated proteins. Human neonatal lungs exposed to fetal tracheal occlusion displayed alveolar deposits of collagen and myofibroblasts. In human CDH as well as in the rabbit model of CDH, tracheal occlusion enhanced the pulmonary expression of transforming growth factor-β (TGFβ) and Rho kinase−associated proteins to the detriment of activation of SMAD2/3, which is normally detected in human lungs with advancing gestation. As an alternative to tracheal occlusion, pulmonary distension by perfluorocarbon in the fetal rabbit model of CDH improved lung mechanics and alveolar elastogenesis without transcriptional changes in extracellular matrix, surfactant protein genes or TGFβ. Finally, intratracheal instillation of BAY 41–2272 in the rabbit fetuses with CDH improved hemodynamics, reduced medial hypertrophy of pulmonary arterioles, and increased capillary bed formation by stimulating endothelial cell proliferation.CONCLUSIONS. In the fetal rabbit model of CDH, poor lung function after tracheal occlusion is compatible with activation of TGFβ and imbalance in extracellular matrix and epithelial homeostasis. In human CDH newborns treated by fetal tracheal occlusion, changes in the pulmonary interstitium and impaired TGFβ signaling raise the question of disturbances of postnatal lung development induced by tracheal occlusion. As potential alternatives to tracheal occlusion, prenatal perfluorocarbon improves lung hypoplasia, whereas prenatal BAY 41–2272 attenuates pulmonary hypertension. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

L’océan Austral est soumis à la redistribution globale des contaminants par les voies atmosphérique et océanique. Cependant, la contamination des écosystèmes austraux est très peu connue, en particulier dans le secteur Indien. De par leur toxicité, leur mobilité et leur capacité à se bioaccumuler dans les tissus des organismes et à se bioamplifier dans les réseaux trophiques, le mercure (Hg) et les polluants organiques persistants (POPs) comptent parmi les contaminants les plus préoccupants. Du fait de leur position élevée dans les réseaux trophiques, les oiseaux marins sont exposés à de grande quantités de contaminants par la voie alimentaire. En conséquence, ils sont souvent utilisés comme bioindicateurs de l’état de contamination des écosystèmes, par le biais des plumes et du sang, qui peuvent être échantillonnés de façon non destructive. Ma thèse s’est intéressée aux nombreuses espèces d’oiseaux marins (plus de 40) qui nichent au sein des Terres Australes et Antarctiques Françaises, au sud de l’Océan Indien, afin de décrire et expliquer les niveaux de contaminants le long d’un large gradient latitudinal, de l’Antarctique à la Zone Subtropicale, et d’identifier les meilleures espèces bioindicatrices pour un suivi à long terme de la contamination de ces écosystèmes. Au cours d’une première étape méthodologique, les manchots et les poussins de toutes les espèces ont été identifiés comme de bons bioindicateurs de contamination puisque, à la différence de la plupart des oiseaux adultes, ils présentent une faible variabilité des niveaux de contaminants dans les plumes. Au cours d’une seconde étape explicative, l’effet de facteurs intrinsèques (traits individuels) et extrinsèques (écologie alimentaire déduite grâce à la méthode des isotopes stables) sur les niveaux de contaminants a été évalué dans les plumes des oiseaux de la communauté de Kerguelen (27 espèces) et dans le sang du grand albatros de Crozet (180 individus dont les traits de vie sont connus). L’écologie alimentaire s’est avérée être le principal facteur explicatif des niveaux de contaminants, tandis que l’âge, le sexe, la phylogénie et le statut reproducteur jouent un rôle mineur. La classe d’âge est néanmoins un facteur à prendre en compte, puisque les poussins montrent souvent des concentrations inférieures aux adultes. Au cours d’une troisième étape, les variations spatio-temporelles de la contamination ont été étudiées en utilisant une sélection d’espèces bioindicatrices et en tenant compte de leur écologie alimentaire. Plusieurs résultats portant sur différentes espèces (oiseaux océaniques) et populations (poussins de skua) ont montré que, contrairement aux prédictions, l’exposition des oiseaux au Hg augmente graduellement des eaux antarctiques aux eaux subantarctiques puis aux subtropicales, alors que l’exposition aux POPs, en accord avec la théorie de la distillation globale, montre la tendance inverse. D’autre part, la comparaison des concentrations en Hg dans les plumes de manchot, effectuée entre des spécimens de musée et des échantillons actuels, indique que leur exposition au Hg n’a pas changée depuis les années 1950-1970. Toutefois, des espèces subantarctiques montrent une tendance à la hausse. De futures études devraient viser à l’utilisation des plumes comme tissu de référence pour l’évaluation et le suivi de la contamination des écosystèmes, en particulier en ce qui concerne les POPs. Parmi les nombreuses espèces étudiées au cours de ces travaux de thèse, les bioindicateurs les plus pertinents se révèlent être le manchot empereur et le pétrel des neiges (Antarctique), le manchot royal, le pétrel bleu et l’albatros à sourcil noirs (subantarctique), le gorfou sauteur subtropical et l’albatros à bec jaune (subtropical). Le suivi à long terme de ces espèces permettra d’évaluer l’évolution temporelle de l’état de contamination de l’océan Austral. / Antarctic and subantarctic marine environments are reached by inorganic and organic contaminants through ocean circulation and atmospheric transport. Yet, environmental contamination is poorly known in the Southern Ocean, in particular in the Indian sector. Among environmental contaminants, mercury (Hg) and persistent organic pollutants (POPs) are primarily of concern, because they are toxic, highly mobile, and they bioaccumulate in the tissues of living organisms and biomagnify up the food web. Seabirds, as upper predators, are exposed to large quantities of contaminants via food intake and have widely served as biomonitors of marine contamination, notably through the non-destructive sampling of their feathers and blood. My doctoral work has focussed on the abundant and diverse seabird species (more than 40) breeding in the French Southern and Antarctic Lands, southern Indian Ocean, in order to describe and explain contaminant concentrations over a large latitudinal gradient, from Antarctica to the subtropics, and to identify the best bioindicator species for contaminant biomonitoring. In a first methodological step, seabirds with synchronous moult of body feathers (adult penguins and chicks of all species) were recognised as good candidates as bioindicators, because, unlike most adult birds, they present low within-individual variation in feather contaminant concentrations. In a second explanatory step, the influence of intrinsic (individual traits) and extrinsic factors (feeding ecology inferred from the stable isotope method) driving variation in contaminant concentrations was evaluated in feathers of the large avian community of the Kerguelen Islands (27 species) and in blood of wandering albatrosses from the Crozet Islands (180 birds of known individual traits). Feeding ecology was the main factor driving variation in contaminant concentrations of blood and feathers, both at the community, population and individual levels, whereas age, sex, phylogeny and breeding status played a minor role. Age-class was however an important intrinsic factor to consider, with chicks usually having lower concentrations than adults. In a third step, spatio-temporal patterns of contamination were studied through selected bioindicator species and by taking into account their feeding habits. Results from different species (oceanic seabirds) and populations (skua chicks) showed that, contrary to predictions, Hg exposure gradually increases from Antarctic to subantarctic and subtropical waters, whereas, in accordance with the global distillation theory, POPs exposure has the opposite pattern. Comparisons between penguin feathers from museum collections and contemporary samples showed that bird exposure to Hg is overall not different today when compared to 50-70 years ago, but subantarctic species are possibly experiencing an increasing trend. Future research efforts should be focussed on the use of feathers as biomonitoring tools, in particular for POPs determination. The best recommended bioindicator species include the emperor penguin and snow petrel (Antarctic), king penguin, blue petrel and black-browed albatross (subantarctic), and northern rockhopper penguin and Indian yellow-nosed albatross (subtropical). Future biomonitoring studies on these species will give invaluable insights into the poorly-known temporal trends of environmental contamination in the Southern Ocean.

Abstract In Finland there is a food monitoring program which has found elevated dioxin and polychlorinated biphenyl concentrations in the muscle of semi-domesticated reindeer (Rangifer tarandus tarandus) calves. This led to further research on the concentrations of persistent organic pollutants in reindeer muscle, liver, and other internal organs. The research was further expanded on wild moose (Alces alces) muscle and liver. The main objective of this thesis is to increase knowledge of polychlorinated dibenzo-p-dioxin (PCDD), polychlorinated dibenzofuran (PCDF), polychlorinated biphenyl (PCB), and polybrominated diphenyl ether (PBDE) pollution levels in the Finnish terrestrial environment, and in semi-domesticated reindeer and wild moose in particular. The research gives information of exposure conditions in the reindeer’s food chain, as well as species differences and individual variation in accumulation and distribution of pollutants in reindeer and moose. Local differences between the contaminant concentrations were explored. Toxic equivalencies (TEQs) set by the World Health Organization (WHO) of PCDD/Fs and PCBs were calculated in order to assess the validity of selling reindeer and moose tissue. It was observed that there is a species-, individual-, and tissue-specific accumulation of dioxins, dioxin-like PCB, and PBDE compounds in reindeer and moose. Varying exposure conditions mainly explain the differences, although taking into account the age of an individual animal, its metabolic patterns have a role, too. Reindeer placenta and milk proved to be important factors in the transporting of compounds from hind to calf. The highest PCDD/F and PCB concentrations (as WHO-TEQs) were observed in reindeer calves in the study area in which animals are fed in natural pastures only. Despite the findings, it was concluded that it is safe to eat reindeer and moose meat since the concentrations of dioxins and dioxin-like PCBs in the muscle are relatively low, and because of the low fat content in these animals. Reindeer liver, for its part, had quite a lot of dioxin-like compounds that may compromise its safety as food, at least on a regular basis. The study shows that Finnish semi-domesticated reindeer and wild moose are good indicator species of POP contamination in a terrestrial environment, reindeer describing the situation in northern parts of the country especially. / Tiivistelmä Suomalaisessa ruokamonitorointitutkimuksessa löydettiin kohonneita dioksiinien ja polykloorattujen bifenyylien pitoisuuksia puolivillin poron vasojen lihaksista. Se johti lisätutkimuksiin pysyvien orgaanisten yhdisteiden pitoisuuksista poron lihaksessa, maksassa ja muissa sisäelimissä. Tutkimus laajeni koskemaan myös hirven (Alces alces) lihasta ja maksaa. Väitöskirjatyö lisää tietämystä polykloorattujen dibentso-p-dioksiinien (PCDD), polykloorattujen dibentsofuraanien (PCDF), polykloorattujen bifenyylien (PCB) ja polybromattujen difenyylieettereiden (PBDE) pitoisuuksista suomalaisessa maaympäristössä ja erityisesti porossa ja hirvessä. Tutkimus antaa tietoa yhdisteille altistumisesta sekä viitteitä lajien- ja yksilöiden välisistä eroavai¬suuksista yhdisteiden kertymisessä ja niiden jakaantumisessa eri kudosten välillä. Alueellista vaihtelua yhdisteiden pitoisuuksissa selvitettiin myös. Maailman terveysjärjestön (WHO) asettamia PCDD/F- ja PCB-yhdisteiden toksisuusekvivalenttiarvoja (TEQ) tarkasteltaessa pystyttiin arvioimaan kemiallista elintarvikekelpoisuutta suhteessa EU:n antamiin sallittuihin pitoisuuksiin. Työssä havaittiin laji-, yksilö-, ikä- ja kudosspesifistä dioksiinien, dioksiinienkaltaisten PCB- ja PBDE-yhdisteiden kerääntymistä porossa ja hirvessä. Tämä on todennäköisimmin seurausta vaihtelevasta altistumisesta yhdisteille, mutta myös lajien metabolisissa toiminnoissa yksilön eri ikäkausina voi olla eroavaisuuksia. Poron istukan ja maidon havaittiin olevan tärkeitä yhdisteiden kulkeutumisessa emolta sikiölle ja vasalle. Korkeimmat dioksiinien ja dioksiinien kaltaisten PCB-yhdisteiden konsentraatiot (WHO-TEQ-pitoisuuksina) havaittiin poron vasoilla tutkimusalueella, jossa eläimet olivat laiduntaneet ainoastaan luonnonlaitumilla. Huolimatta havaituista haitta-ainepitoisuuksista todettiin, että sekä luonnostaan vähärasvaisen poron että hirvenlihan syönti on turvallista suhteellisen alhaisten dioksiinien ja dioksiininkaltaisten PCB-yhdisteiden pitoisuuksien perusteella. Poron maksa puolestaan sisälsi melko korkeita dioksiinien kaltaisten yhdisteiden pitoisuuksia, mikä voi vaikuttaa sen turvalliseen käyttöön elintarvikkeena ainakin usein syötynä. Suomalainen puolivilli poro ja villi hirvi sopivat hyvin POP-kontaminoitumisen indikaattoreiksi maaympäristössä; poron erityisesti kuvaten tilannetta maan pohjoisosissa.

Le gyrus denté occupe une position clé au sein du lobe temporal des mammifères en constituant le point de contrôle entre le système néocortical et le système hippocampique. Considéré comme la porte de l'hippocampe, le gyrus denté filtre les activités excitatrices en provenance du cortex entorhinal grâce à la décharge éparse des cellules granulaires. Ce type de codage neuronal lui confère également un rôle déterminant dans les mécanismes de l'apprentissage et de la mémoire lors de la distinction d'évènements similaires mais différents, en permettant la décorrélation des patrons d'activité corticale. Grâce à un ensemble de propriétés structurales et fonctionnelles, les cellules granulaires du gyrus denté génèrent des évènements synaptiques extrêmement rapides restreignant leur fenêtre temporelle d'intégration et leur permettant de jouer le rôle de détecteurs de coïncidence. Au cours d'une épilepsie du lobe temporal (ELT), l'hippocampe présente d'importantes altérations de codage neuronal qui pourraient participer aux troubles cognitifs décrits chez les patients et les modèles animaux. Dans ces conditions pathologiques, les axones des cellules granulaires du gyrus denté (les fibres moussues) bourgeonnent et établissent des synapses aberrantes au niveau d'autres cellules granulaires, créant ainsi un puissant réseau excitateur récurrent. Ces fibres moussues récurrentes convertissent la nature de la transmission glutamatergique car elles opèrent via des récepteurs kaïnate générant des potentiels post-synaptiques à cinétique lente, absents en condition contrôle. / The dentate gyrus plays a major role at the gate of the hippocampus, filtering incoming information from the entorhinal cortex. A fundamental coding property of dentate granule cells (DGCs) is their sparse firing. Indeed, they behave as a coincidence detector due to the fast kinetics of excitatory synaptic events restricting integration of afferent inputs to a narrow time window. In temporal lobe epilepsy (TLE), the hippocampus displays important coding alterations that may play a role in cognitive impairments described in patients and animal models. However, the cellular mechanisms remain poorly understood. In animal models of TLE and human patients, neuronal tissue undergoes major reorganization; some neurons die whereas others, which are severed in their inputs or outputs, sprout and form novel aberrant connections. This phenomenon, called reactive plasticity, is well documented in the dentate gyrus where DGC axons (the mossy fibres) sprout and create a powerful excitatory network between DGCs. We recently showed that in addition to the axonal rewiring, recurrent mossy fibres convert the nature of glutamatergic transmission in the dentate gyrus because they operate via long-lasting kainate receptor (KAR)-mediated EPSPs (EPSPKA) not present in the naive condition.

La spasticité est l’une des nombreuses complications motrices qui peuvent apparaître après une lésion de la moelle épinière. Elle est présente dans 75 % des patients médullo-lésés et se caractérise par une hypertonie musculaire en réponse à un réflexe d’étirement. Les traitements actuels, qui ciblent les symptômes et non les causes de la spasticité, sont peu efficaces. Bien que les mécanismes neurologiques qui sous-tendent la spasticité soient complexes et restent en grande partie méconnus, un certain consensus se dégage sur le fait qu’elle est associée à une hyperexcitabilité intrinsèque des motoneurones et à une levée de l’inhibition des réflexes spinaux. L’hyperexcitabilité motoneuronale se manifeste par une décharge soutenue de potentiels de plateau et résulte en partie d’une augmentation des courants entrants persistants sodiques (INaP). La désinhibition découle, en partie, d’une baisse de l’expression des cotransporteurs potassium-chlorure de type 2 (KCC2) à la membrane des motoneurones, modifiant ainsi le gradient électrochimique des ions Cl- et donnant un caractère excitateur aux deux principaux neurotransmetteurs inhibiteurs que sont le GABA et la glycine. Néanmoins, les mécanismes à l’origine des dérégulations du courant INaP et des co-transporteurs KCC2 ne sont toujours pas élucidés. / Spasticity is commonly caused by several pathologies and specifically after a spinal cord injury (SCI). Spasticity is usually associated with hypertonia, clonus, muscle spasm and pain. The present thesis aims to identify the upstream mechanism in the pathophysiology of spasticity Calpain, a calcium-activated cysteine protease, has been shown to participate in the development of the inflammatory processes after SCI. Of special interest, some determinants governing the inactivation of sodium (Na+) channels are sensitive to proteases and their proteolytic cleavage prevents inactivation of Na+ channels. As a result, INaP is strongly increased. It is worth mentioning that the C-terminal domain of KCC2 is also sensitive to proteases which alter KCC2 ability to extrude Cl- ions. Among the different proteases, calpains are able to truncate both Na+ channels and KCC2 transporters. This led us to consider the exciting possibility that a proteolytic cleavage of both Na+ channels and KCC2 by calpains could compose an upstream inflammatory mechanism contributing to the development of spasticity after SCI. My thesis demonstrates that the cleavage of Na+ channels and KCC2 by calpain after SCI, is responsible for the upregulation of INaP and disinhibition of motoneurons, that both act synergistically to generate spasticity. Calpain inhibition by MDL28170 reduced the cleavage of both Na+channels and KCC2 associated with a respective downregulation of INaP, hyperpolarizing shift of the EIPSP, and an alleviation of spasticity. The thesis represents a significant breakthrough by opening novel perspectives to develop therapies.