Ce travail porte sur la modélisation et la simulation du trafic routier sur un réseau. Modéliser le trafic sur une section homogène (c'est-à-dire sans entrée, ni sortie) trouve ses racines au milieu du XXème siècle et a généré une importante littérature depuis. Cependant, la prise en compte des discontinuités des réseaux comme les jonctions, n'a attiré l'attention du cercle scientifique que bien plus récemment. Pourtant, ces discontinuités sont les sources majeures des congestions, récurrentes ou non, qui dégradent la qualité de service des infrastructures. Ce travail se propose donc d'apporter un éclairage particulier sur cette question, tout en s'intéressant aux problèmes d'échelle et plus particulièrement au passage microscopique-macroscopique dans les modèles existants. La première partie de cette thèse est consacrée au lien existant entre les modèles de poursuite microscopiques et les modèles d'écoulement macroscopiques. Le passage asymptotique est assuré par une technique d'homogénéisation pour les équations d'Hamilton-Jacobi. Dans une deuxième partie, nous nous intéressons à la modélisation et à la simulation des flux de véhicules au travers d'une jonction. Le modèle macroscopique considéré est bâti autour des équations d'Hamilton-Jacobi. La troisième partie enfin, se concentre sur la recherche de solutions analytiques ou semi-analytiques, grâce à l'utilisation de formules de représentation permettant de résoudre les équations d'Hamilton-Jacobi sous de bonnes hypothèses. Nous nous intéressons également dans cette thèse, à la classe générique des modèles macroscopiques de trafic de second ordre, dits modèles GSOM / This work focuses on modeling and simulation of traffic flows on a network. Modeling road traffic on a homogeneous section takes its roots in the middle of XXth century and it has generated a substantial literature since then. However, taking into account discontinuities of the network such as junctions, has attracted the attention of the scientific circle more recently. However, these discontinuities are the major sources of traffic congestion, recurring or not, that basically degrades the level of service of road infrastructure. This work therefore aims to provide a unique perspective on this issue, while focusing on scale problems and more precisely on microscopic-macroscopic passage in existing models. The first part of this thesis is devoted to the relationship between microscopic car-following models and macroscopic continuous flow models. The asymptotic passage is based on a homogenization technique for Hamilton-Jacobi equations. In a second part, we focus on the modeling and simulation of vehicular traffic flow through a junction. The considered macroscopic model is built on Hamilton-Jacobi equations as well. Finally, the third part focuses on finding analytical or semi-analytical solutions, through representation formulas aiming to solve Hamilton-Jacobi equations under adequate assumptions. In this thesis, we are also interested in a generic class of second order macroscopic traffic flow models, the so-called GSOM models

La Sclérose Latérale Amyotrophique (SLA) est une affection neurodégénérative touchant les motoneurones, habituellement mortelle en 3 à 5 ans. La cause de la maladie n'est pas connue et le seul traitement actuellement disponible ne permet qu'un allongement modeste de la survie. Des altérations fonctionnelles de la jonction neuromusculaire (JNM) ont été rapportées dans la SLA mais leur origine physiopathologique n'est pas connue. Nous avons étudié les JNM chez 11 patients atteints de SLA, en associant étude morphologique en microscopie confocale et analyse ultrastructurale. L'analyse fonctionnelle réalisée en EMG de surface retrouvait une anomalie de transmission neuromusculaire (décrément > 10%) chez 45% des patients. Des altérations morphologiques des JNM étaient visibles chez tous les patients, y compris au stade précoce de la maladie. Associé aux anomalies en rapport avec le phénomène de dénervation, nous avons observé un aspect anormal de spiculation de la gouttière primaire dans environ un tiers des cas. Une interposition marquée de la cellule de Schwann terminale entre la terminaison nerveuse et la membrane postsynaptique, pouvant altérer la transmission synaptique, était parfois visible. Nous avons objectivé une réinnervation compensatrice significativement plus importante chez les patients présentant une SLA d'évolution lente et montré que certains facteurs moléculaires musculaires comme l'histone déacétylase 4 pourraient jouer un rôle crucial dans la capacité de réinnervation. Ce travail a mis en évidence des altérations morphologiques majeures au niveau des JNM des patients atteints de SLA et a permis d'identifier des cibles thérapeutiques potentielles. / Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disorder affecting motor neurons, usually leading to death in 3 to 5 years. The only treatment currently available, riluzole, has a modest effect on survival. Functional alterations of the neuromuscular junction (NMJ) have been reported in ALS, but their pathophysiological significance remains unknown. We studied the morphology of neuromuscular junctions in muscle samples collected from 11 ALS patients, using confocal and electron microscopy. Functional analysis of the NMJs was performed using surface-recording of compound motor action potentials after repetitive nerve stimulation at slow stimulus rate. A significant decrement (>10%), suggesting impairment of the neuromuscular transmission, was present in 45% of the patients. Morphological alterations of the NMJs were present in all ALS patients even at the early-stages. Beside denervation-induced morphological changes, one third of the NMJs showed abnormal spike-like areas of the outer edge of the postsynaptic primary gutter. A marked interposition of the terminal Schwann cell between the nerve terminal and the postsynaptic membrane, which was likely to alter synaptic transmission, was sometimes present. We found a significantly greater compensatory reinnervation in muscle from patients with slowly progressive ALS. Furthermore, we identified that the muscle molecular factor histone deacetylase 4 could play a key role in muscle reinnervation and disease progression in patients with ALS. This work has highlighted the presence of major morphological changes at the NMJs of ALS patients and identified potential new targets for future treatment.

La sclérose latérale amyotrophique (SLA) est une maladie neurodégénérative dont les premiers symptômes apparaissent généralement vers 60 ans. Elle affecte sélectivement le système moteur et provoque une paralysie progressive amenant au décès du patient par défaillance respiratoire en quelques années. À ce jour, il n’existe aucun traitement curatif, d’où la nécessité de comprendre la physiopathologie de la SLA. Bien que de nombreuses altérations dans le muscle aient été mises en évidence, sa contribution dans la SLA reste à définir. Nous avons montré que FUS est enrichi dans les noyaux sous-synaptiques de façon dépendante de l’innervation. De plus, FUS se lie au promoteur des récepteurs de l’acétylcholine et induit leur transcription de façon dépendante d’ERM. Le mutant FUS, quant à lui, est enrichi dans les noyaux extra-synaptiques et entraîne une toxicité musculaire responsable de l’altération de la jonction neuromusculaire (JNM). Au-delà de la JNM, FUS active MEF2A, de façon dépendante de PRMT1 afin de réguler les fonctions mitochondriales et la différenciation musculaire. La toxicité musculaire de FUS joue donc un rôle clé dans la physiopathologie de la SLA. / Amyotrophic lateral sclerosis is a neurodegenerative whose first symptoms generally appear around age 60. It is characterized by progressive motor neuron degeneration, paralysis and leading to death due to respiratory failure in a few years. Currently, there is no cure so the understanding of ALS physiopathology is necessary. Although many alterations in the muscle have been highlighted, its contribution in ALS remains to be defined. We showed that FUS is enriched in subsynaptic nuclei and this enrichment depended on innervation. Besides, FUS binds directly acetylcholine receptors (AchR) promoter and is required for Ermdependent induction of AChR expression. Conversely, mutant FUS is enriched on extra-synaptic nuclei and induce muscle intrinsic toxicity responsible for neuromuscular junction (NMJ) alteration. Beyond NMJ, FUS is required for muscle mitochondrial function and muscle differentiation through PRMT1-dependent MEF2A activation. Thus, FUS muscular toxicity plays a key role in the ALS physiopathology.

La bande transporteuse est utilisée dans l’industrie pour acheminer de la matière ou des objets dans de multiples domaines d’applications tels que les mines souterraines ou aériennes, les carrières, l’industrie alimentaire, l’industrie agricole ou les supermarchés. Ses composants lui permettent de travailler dans des conditions extrêmement difficiles. L’intérêt est porté sur les bandes transporteuses de mines de fond dont les contraintes de fonctionnement et d’acheminement sont particulières. En effet, la bande est acheminée sur rouleaux et jonctionnée sur le site de l’utilisateur pour former une bande sans fin. La jonction est le point faible de la bande transporteuse car sa résistance représente 50% seulement de la force de rupture nominale de la bande. L’objet de cette étude est de proposer une nouvelle solution de jonctionnement.Après avoir analysé les différentes contraintes mécaniques et chimiques auxquelles la bande transporteuse est soumise, une étude comparative sur les différents types de jonctions a été menée.Il y a les jonctions chimiques et les jonctions mécaniques. Chacune a des avantages et des inconvénients en termes de coût ou de longévité, qui ont été présentés. Dans un troisième temps, une étude a été menée sur la compréhension du comportement de rupture de la jonction mécanique. L’étude sur la jonction a été réalisée à l’aide d’un montage expérimental conçu spécifiquement. L’influence des paramètres de la jonction a pu être évaluée grâce à un nouvel indicateur adimensionnel représentant l’efficacité de la jonction par rapport à la bande normale (Junction Tensile Strength Efficiency : JTSE). Dans un quatrième temps, une étude a été faite sur un système de jonction innovant par couture. La jonction réalisée a été testée sur un banc de traction pour en vérifier l’efficacité, puis des essais ont été effectués sur un banc dynamique pour en étudier la fatigue. La jonction cousue est une innovation qui permet de dégager de nouvelles perspectives dans le jonctionnement des bandes transporteuses. / The conveyor belt is used as a material and object transportation tool in a lot of industries such as underground mines, quarries, food industries, agribusinesses or supermarkets. The studied subject is the underground mine conveyor belt, because of their transportation and use difficult conditions.Indeed, the conveyor belt must be cut to be dispatched using belt reeling and joined afterwards in the underground mine forming an endless belt. However, the junction is the weakest part of the conveyor belt due to the 50% belt weakening. The aim of this study is to propose a new junction solution. After analysing the underground belt mechanical and chemical use conditions, the different junction types have been analysed. A junction advantage and inconvenience comparison has been presented. Then, the junction mechanical behaviour has been studied using an original mechanical setup reproducing the mechanical junction. The different parameter influence has been evaluated using a new adimensional indicator called junction tensile strength efficiency. In the last part, a new junction solution made by a sewn has been explored. The sewn junction tensile efficiency has been tested on a tensile strength machine and its fatigue efficiency has been verified through dynamic tests. New outlines in the conveyor belt joining are open as a result of the sewn junction solution development.

Le développement des systèmes de conversion photovoltaïques ces trente dernières années a permis des améliorations considérables en terme de coût et de performances. A ce jour, les meilleurs rendements de conversion photovoltaïques sont obtenus avec des systèmes à oncentration solaire utilisant des cellules multi-jonctions (MJ) à base de matériaux semi-conducteurs III-V. Dans ce domaine, le meilleur rendement atteint à ce jour est de 46,0 % sous une concentration de 508 soleils avec une cellule à 4 jonctions issu du partenariat Soitec/Fraunhofer ISE/CEA. Cette cellule MJ est composée d’une cellule tandem accordée sur GaAs assemblée par collage moléculaire à une autre cellule tandem accordée sur InP. Bien que le rendement atteint soit élevé, les performances de la cellule sont limitées sous fortes concentrations à cause de ce collage moléculaire. Dans le domaine des fortes concentrations, le record est actuellement détenu par la société américaine Solar Junction avec un rendement de 44,0 % mesuré sur une cellule triple jonction monolithique en GaInP/GaAs/GaInNAs de 0,3 cm² pour un taux de concentration de 942 soleils (irradiance directe de 942 kW/m²). Une seconde cellule a atteint un rendement performant à une irradiance directe supérieure à 1 MW/m², il s’agit d’une cellule tandem en GaInP/GaAs de l’IES-UPM qui a atteint 32,6 % sous une concentration de 1026 soleils.Dans le contexte précédent, les travaux présentés dans ce manuscrit visent à l’évaluation d’une nouvelle filière dans le domaine du CPV à base de semi-conducteurs III-V : la filière antimoniure (III-Sb). Les cellules que nous avons étudiées dans le cadre de cette thèse sont à base de GaSb et de l’alliage AlxGa1-xAsySb1-y, fabriquées de façon monolithique par MBE (Molecular Beam Epitaxy) sur substrat GaSb. Ce type de cellules, du fait de la très bonne complémentarité des gaps des matériaux, constitue une alternative crédible et originale aux cellules existantes pour une utilisation sous flux solaire fortement concentré.Le travail à réaliser dans le cadre de cette thèse porte sur :- La caractérisation électrique et optique des alliages quaternaires utilisés.- La conception et le design des cellules.- La réalisation et la mise au point de toutes les étapes technologiques nécessaires à la conception des cellules (photolithographie UV, gravure, métallisation, …).- La caractérisation électrique et optique des cellules fabriquées (I(V), TLM, réponse spectrale, …).- La caractérisation des cellules sous flux solaire (fortement) concentré.Ce travail a été cofinancé par l’Université de Montpellier et le LabEx SOLSTICE. / The development of photovoltaic conversion systems these past thirty years led to considerable improvements in terms of cost and performances. The best conversion efficiencies are currently obtained with solar concentration systems associated with multi-junction solar cells (MJSC) made of III-V materials. In this field, the record efficiency is of 46.0% under a 508-sun solar concentration with a 4-junction cell from Soitec/Fraunhofer ISE/CEA. This MJSC is composed of a tandem cell lattice-matched to GaAs wafer bonded to another tandem cell lattice-matched to InP. Although it reached high conversion efficiency, its performances are limited under solar concentration because of the wafer bonding. In the field of high solar concentrations, the record is held by Solar Junction with a monolithic triple junction GaInP/GaAs/GaInNAs cell of 0.3 cm² that reached an efficiency of 44.0% under 942 suns (direct irradiance of 942 kW/m²). Another high solar concentration efficiency record worth mentioning is held by IES-UPM with a tandem solar cell (GaInP/GaAs) that reached an efficiency of 32.6% under a concentration of 1026 suns.In this context, the work presented in this manuscript aims to evaluate the potential of a new family of III-V materials for high solar concentration applications: antimonide-based materials (III-Sb). The studied cells in this thesis are made out of GaSb and the quaternary AlxGa1-xAsySb1-y, monolithically grown by MBE (Molecular Beam Epitaxy) on a GaSb substrate. These materials, thanks to the large range of available band-gaps, represent an original and well-founded alternative to existing solar cells for high solar concentration applications.The work achieved in this thesis covers:- The electrical and optical characterization of the quaternary materials used.- The conception and designing of the cells.- The production and tuning of every technological steps in order to fabricate our solar cells (UV photolithography, etching, metal deposition,…).- The electrical and optical characterization of our fabricated solar cells (I(V), TLM, spectral response,…).- The characterization under (high) solar concentration of our cells.This work was cofounded by the University of Montpellier and the LabEx SOLSTICE.

INTRODUÇÃO: A técnica de eletromiografia de fibra única (EMGFU), mediante análise do jitter, é o método neurofisiológico mais sensível para a confirmação do distúrbio da junção neuromuscular na miastenia gravis (MG). Os registros são tradicionalmente obtidos com agulha de fibra única, de alto custo e reutilizável. Por causa da necessidade atual do uso de material descartável, a agulha concêntrica vem sendo utilizada em substituição à agulha de fibra única. A técnica utilizada é semelhante, porém os potencias de ação para a análise do jitter são obtidos com eletrodo de agulha concêntrica (Eletromiografia de fibra única - jitter com agulha concêntrica, EMGFU-JAC). Contudo, os estudos são escassos e as metodologias utilizadas são heterogêneas com a utilização dessa agulha. OBJETIVOS: Este estudo tem por objetivo mensurar os valores de jitter obtidos com agulha concêntrica, no músculo Orbicularis Oculi, em sujeitos saudáveis e em pacientes com MG autoimune adquirida e avaliar a validade do método nas formas generalizada e ocular da doença. MÉTODOS: Foram estudados 20 sujeitos saudáveis, 20 pacientes com miastenia gravis forma generalizada (grupo MGG) e 13 com a forma ocular da doença (grupo MGO). A EMGFU-JAC foi realizada em todos os participantes, idealmente com 20 medidas de jitter em cada estudo. O jitter foi expresso como a média das diferenças consecutivas (MCD). Em todos os pacientes do estudo foram realizados o teste de estimulação repetitiva e dosagem sérica de anticorpo antirreceptor de acetilcolina (ac-AChR) no momento da análise do jitter. Nos pacientes soronegativos para ac-AChR, foi pesquisado o anticorpo antimúsculo específico tirosina-quinase (ac-MuSK). Foram definidos o limite superior da normalidade (LSN) para a média do MCD de cada estudo e para valores individuais de MCD. Os critérios de anormalidade foram: (1) média do MCD acima do LSN; ou (2) mais de 10% dos valores individuais de MCD acima do LSN. A definição do LSN para valores individuais de MCD baseou-se no conceito de que dois entre 20 valores de MCD acima do LSN são aceitáveis em um músculo saudável, para a técnica de contração voluntária. Portanto, estimou-se o LSN para o 18o valor mais alto de MCD (18o par). Para análise da acurácia do método, foram construídas duas curvas ROC (Receiver Operating Characteristic) para as variáveis média do MCD e 18o par, no grupo de pacientes (MGG e MGO) versus controle. RESULTADOS: No grupo controle a média das médias do MCD foi (19,0 ± 2,4)us e a média do 18o valor mais alto de cada estudo foi (24,5 ± 3,6)us. Esses valores obtidos apresentaram distribuição Gaussiana e o LSN foi definido como a média desses valores + 2 DP. O LSN para a média do MCD foi 24us, e 32?s para valores individuais de MCD. No grupo MGG, a análise do jitter foi anormal em todos os 20 pacientes por ambos os critérios de anormalidade, exceto em um paciente que apresentou anormalidade por apenas um dos critérios. No grupo MGO, apenas um dos 13 pacientes não preencheu os critérios de anormalidade. No grupo de pacientes, a positividade da EMGFU-JAC foi maior do que o teste de estimulação repetitiva e dosagens de anticorpos. Nas curvas ROC para as variáveis médias do MCD e 18o par, o valor de melhor sensibilidade (93,9%), sem resultados falsos positivos, foi 24,7us e 33,1us, respectivamente. CONCLUSÕES: A EMGFU-JAC apresenta alta sensibilidade e especificidade na identificação de distúrbio da transmissão neuromuscular em pacientes com MG. A utilização da agulha concêntrica é válida para a análise do jitter, como alternativa à agulha de fibra única / INTRODUCTION: Single fiber electromyography (SFEMG) technique, through jitter analysis, is the most sensitive neurophysiological method for confirmation of neuromuscular junction disorder in myasthenia gravis (MG). Records are traditionally obtained with single fiber needle, which is reusable and has a high-cost. Due to the current need of using disposable material, concentric needle has been used to replace single fiber needle. The technique is similar, but the action potential for jitter analysis is obtained with concentric needle electrode (SFEMG - concentric needle jitter, SFEMG-CNJ). However, studies are scarce and methodologies used are heterogeneous with the use of this needle. OBJECTIVES: This study aims to measure jitter values obtained with concentric needle in the Orbicularis Occuli muscle in healthy subjects and in patients with autoimmune acquired MG and to assess the validity of the method in generalized and ocular forms of the disease. METHODS: 20 healthy subjects, 20 patients with generalized myasthenia gravis (GMG group) and 13 with the ocular form of the disease (OMG group) were studied. SFEMG-CNJ was performed on all participants, ideally with 20 jitter values in each study. Jitter was expressed as the mean consecutive difference (MCD). Repetitive nerve stimulation and serum acetylcholine receptor antibody (AChR-ab) were performed in all patients in the study, by the time of jitter analysis. Tyrosine kinase specific antibody muscle antibodies (MuSK-ab) were performed in AChR-ab negative patients. The upper limit of normality (ULN) for the mean MCD and for individual jitter values were defined. The abnormality criteria were: (1) mean MCD above ULN; or (2) more than 10% of individual jitter values above ULN. The definition of ULN for individual jitter values was based on the concept that two out of 20 jitter values above ULN are acceptable in a healthy muscle for voluntary contraction technique. Therefore, the ULN for the 18th highest jitter value (18 pair) was estimated. To analyze the method\'s accuracy, two ROC curves (Receiver Operating Characteristic) for the mean MCD and 18th pair in the group of patients (MGG and MGO) versus control were constructed. RESULTS: In the control group the mean of MCD means was (19.0 ± 2.4)us and the mean of the 18 highest value of each study was (24.5 ± 3.6)us. These values showed Gaussian distribution and the ULN was set as the mean of these values + 2 SD. The ULN for the mean MCD was 24us, and 32us for individual values of MCD. In GMG group, jitter analyses were abnormal in all 20 patients based on both abnormality criteria, except in one patient, who had abnormalities in only one of the criteria. In OMG group, only one patient from 13 met neither of the abnormality criteria. In patients, the positivity of SFEMG-CNJ was higher than repetitive nerve stimulation test and antibody detection. The ROC curve threshold showing the best sensitivity (93.9%) with no false positive results was 24.7Us for the mean MCD and 33.1us for individual pairs, respectively. CONCLUSIONS: SFEMG-CNJ has high sensitivity and specificity in identifying neuromuscular transmission disorder in patients with MG. The use of concentric needle is valid for jitter analysis as an alternative to single fiber needle

Estudos anteriores de nosso laboratório demonstraram que a estimulação aguda dos receptores 2-adrenérgicos (2-AR) atenua a perda de massa muscular induzida pela desnervação motora (DEN) por meio de uma via dependente de AMPc/PKA. No entanto os mecanismos moleculares envolvidos na ativação crônica destes receptores ainda são pouco conhecidos. Por outro lado, a ativação desta via de sinalização também está envolvida no controle da estabilidade dos receptores nicotínicos (AChR) na junção neuromuscular (JNM), sugerindo que a densidade dos AChR possa estar sob controle neuro-humoral. Desta forma, aventou-se a possibilidade de que além dos efeitos protetores na massa muscular, a ativação dos receptores 2-AR pudesse mediar a estabilização dos AChR na placa motora. Para testar essa hipótese, camundongos foram submetidos à DEN através da secção do nervo ciático, um protocolo clássico de indução de atrofia muscular e desestabilização dos AChR, e tratados com salina ou clembuterol (CB), um 2-agonista seletivo, por até 14 dias. Após 3 dias de DEN, observou-se redução da massa muscular e aumento do conteúdo proteico e expressão do RNAm de genes relacionados à ativação do sistema Ubiquitina-Proteassoma (atrogina-1 e MuRF1) e do sistema autofágico/lisossomal (catepsina L e LC3). A DEN também promoveu aumento no turnover dos AChR, no número de vesículas endocíticas e na expressão do RNAm para a subunidade 1 dos AChR. Após 7 dias, a DEN reduziu a expressão dos genes relacionados à atrofia e aumentou a atividade da via do AMPc/PKA independentemente do tratamento com CB. Na tentativa de elucidar os sinais extracelulares que produziam esta resposta adaptativa, foi demonstrado que neurônios catecolaminérgicos trafegam ao longo do nervo ciático e sua ablação pela DEN reduziu o conteúdo de noradrenalina muscular. Baseados nestes resultados, foi postulado a existência de uma hipersenbilidade às catecolaminas em músculos desnervados cronicamente. O tratamento com CB por 3 dias aboliu o aumento da expressão dos atrogenes induzido pela DEN e este efeito foi associado ao maior conteúdo de AMPc e de substratos fosforilados pela PKA. Além disso, o CB diminuiu a hiperexpressão do RNAm para catepsina L e LC3 induzida pela DEN de 7 dias. Embora o CB não tenha alterado a meia-vida dos AChR em músculos inervados e desnervados, houve um total bloqueio do aumento do número de vesículas endocíticas contendo o AChR em músculos desnervados e tratados com CB. Corroborando estes dados, o CB aumentou a incorporação de AChR novos nas JNM e este efeito foi também associado à maior expressão do RNAm para a subunidade 1-AChR em músculos desnervados. Esta ação do CB no turnover dos AChR parece ser direta uma vez que neuroniôs catecolaminérgicos presentes no nervo ciático ativam receptores 2-ARe a produção de AMPc especificamente na JNM. Em estudos in vitro, foi demonstrado que a estimulação colinérgica produzida pelo carbacol (10-4M) diminuiu a velocidade de síntese de proteínas, aumentou a proteólise total e a atividade do sistema proteolítico Ca2+-dependente em músculos soleus de ratos por meio da ativação dos receptores nicotínicos. Este efeito catabólico do carbacol foi completamente bloqueado pela adição de CB (10-4M) ao meio de incubação. Os dados obtidos no presente estudo permitem sugerir que a estimulação crônica dos 2-AR no músculo esquelético induz um efeito anti-catabólico pela supressão dos sistemas proteolíticos proteassomal e lisossomal, provavelmente através da via de sinalização do AMPc/PKA. A inibição destes sistemas pode estar relacionada ao aumento do turnover dos AChR, uma vez que a velocidade de incorporação destes receptores na JNM foi aumentada pelo CB. Além disso, os achados que mostram a associação entre neurônios noradrenérgicos e colinérgicos no nervo ciático, que conjuntamente inervam as JNM, e a co-localização de receptores 2-AR e AChR na sinapse permitem sugerir a existência de uma interação funcional entre o sistema colinérgico e adrenérgico na manutenção da massa muscular e da placa motora. / Previous studies from our laboratory have shown that the acute stimulation of 2-adrenergic receptor (2-AR) attenuates the muscle loss induced by motor denervation (DEN) through a cAMP/PKA dependent pathway. However, the molecular mechanisms involved in the chronic activation of these receptors are poorly understood. Furthermore, the activation of this signaling pathway is also involved in controlling the stability of nicotinic receptors (AChR) at the neuromuscular junction (NMJ), suggesting that the density of AChR may be under neurohumoral control. Thus, we postulated that besides the protective effects on muscle mass the activation of 2-AR receptors could mediate the stabilization of AChR in the motor plate. To test this hypothesis, mice were submitted to DEN through of the sciatic nerve section, a classical protocol of induction muscle atrophy and destabilization of AChR, and were treated with saline or clenbuterol (CB), a selective 2-agonist for 14 days. DEN decreased the muscle mass and increased the protein content and mRNA expression of genes related to the activation of the ubiquitin-proteasome system (atrogin-1 and MuRF1) and autophagic/lysosomal system (cathepsin L and LC3). DEN also promoted an increase in the turnover of AChR, number of endocytic vesicles and the expression of mRNA for the 1 subunit of AChR. Interestingly, chronic DEN induced down-regulation of atrophy related-genes, and increased the activity of cAMP/PKA pathway independently of CB treatment. In an attempt to elucidate the extracellular signals that produced this adaptive response, it was demonstrated that catecholaminergic neurons travels along the sciatic nerve and its ablation by DEN reduces muscle norepinephrine content. Based on these results, it was postulated the existence of a muscle adrenergic hypersensitivity to circulating catecholamines induced by chronic DEN. CB treatment for 3 days completely abolished the higher expression of atrogenes and this effect was associated with increased Camp content and PKA phosphorylated substrates. Furthermore, CB decreased the DEN-induced hyperexpression of cathepsin L and LC3 mRNA at 7 days. Although CB has not altered the half-life of AChR in innervated and denervated muscles, it produced a total blockage of the increased number of endocytic vesicles containing the AChR in denervated muscles. Consistently, CB increased the incorporation of new AChR and this effect was associated with an increased expression of the 1-subunit AChR mRNA in denervated muscles. This action of CB on AChR turnover appears to be direct, since catecholaminergic neurons are present in the sciatic nerve stimulating 2-AR and cAMP production specifically in the NMJ. Furthermore, in vitro studies demonstrated that cholinergic stimulation produced by carbachol (10-4M) decreased the rate of protein synthesis and increased the proteolytic activity of Ca2+-dependent system in rat soleus muscle through activation of nicotinic receptors. This catabolic effect of carbachol was completely blocked by the addition of CB (10-4M) to the incubation medium. These data suggest that chronic stimulation of 2-AR in skeletal muscle induces an anti-catabolic effect by suppressing proteasomal and lysosomal proteolytic systems, probably through the cAMP/PKA signaling. The inhibition of these systems seems to be related to the increased AChR incorporation into NMJ induced by CB treatment. Moreover, the association between noradrenergic and cholinergic neurons in the sciatic nerve, both of which innervate the motor endplates, and the co-localization of AChR and 2-ARat the synapse suggest the existence of a functional interaction between cholinergic and adrenergic systems in the maintenance of muscle mass and motor endplate.

A vida é um processo dinâmico no tempo e no espaço, resultante da interação celular provida pelas junções comunicantes (JCs), canais formados por conexinas (Cxs) que permitem a passagem de moléculas de até 1kDa, e íons. O foco deste trabalho foi caracterizar a modulação de Cxs no desenvolvimento, adaptação visual e após trauma mecânico localizado na retina, através de técnicas como PCR em tempo real, western blot e imuno-histoquímica. No desenvolvimento, a Cx43 apresentou alta expressão no início (255%, P<0.01), a Cx45 teve alta expressão em todos os estágios, as Cx50 e Cx36 são pouco expressas no início (4-9%, P<0.01), mas muito expressas no final e no adulto. Na adaptação ao escuro houve regulação de 115% do RNam da Cx36 após 24h, evidenciando uma plasticidade do acoplamento frente às condições de iluminação. Na neurodegeneração, a expressão da Cx36 não variou nos diferentes tempos pós-lesão, mas a Cx43 aumentou após 7 dias. O papel do acoplamento na degeneração foi estudado utilizando bloqueadores e abridores de JCs combinados com métodos para avaliar a viabilidade celular (TUNEL, FluoroJade e LDH). Finalmente, o bloqueio das JCs diminuiu a morte celular secundária, mas a abertura gerou um efeito inicial pró-apoptótico que depois de 4h foi diluído no tecido. / Life is a dynamic process in time and space, resulting from an interaction provided by gap junctions (GJs), channels composed of connexins (Cxs) that permits the passage of molecules up to 1kDa and ions. The aim of this study was analyze the modulation of Cxs during development, dark adaptation and after local mechanical trauma in the retina using techniques like real time PCR, western blot and imunohistochemistry. In development, Cx43 was highly expressed in the beginning (255%, P<0.01), Cx45 was highly expressed during all development, Cx50 and Cx36 were virtually absent in the beginning (4-9%, P<0.01), but highly expressed in the end of development. In Dark-adaptation, Cx36 was up-regulated after 24h (115%, P < 0.05) suggesting a plasticity of the neuronal coupling in response to light conditions. In neurodegeneration, Cx36 expression didnt ranged in different days after lesion, but the Cx43 increased after 7 days. The coupling was studied by using GJ blockers and openers combined with methods to evaluating cellular viability (TUNEL, FluoroJade and LDH). Finally, blocking the cellular coupling reflects in a decrease of secondary death. GJ openers seems act allowing the spread of death in the beginning, but this effect be diluted after 4h in the tissue.

Single-photon sources are essential building blocks in quantum photonic networks, where quantum-mechanical properties of photons are utilised to achieve quantum technologies such as quantum cryptography and quantum computing. In this thesis, a single-photon source driven by a surface acoustic wave (SAW) is developed and characterised. This single-photon source is based on a SAW-driven lateral n-i-p junction in a GaAs quantum-well structure. On this device, the lateral n-i-p junction is formed by gate-induced electrons and holes in two adjacent regions. The SAW potential minima create dynamic quantum dots in a 1D channel between these two regions, and are able to transport single electrons to the region of holes along the channel. Single-photon emission can therefore be generated as these electrons consecutively recombine with holes. After characterisation and optimisation in four batches of devices, clear SAW-driven charge transport and the corresponding electroluminescence (EL) can be observed on an optimised SAW-driven n-i-p junction. Time-resolved measurements have been carried out to study the dynamics of SAW-driven electrons. Time-resolved EL signals indicate that a packet of electrons is transported to the region of holes in each SAW minimum. In addition, the carrier lifetime of SAW-driven electrons in the region of holes is shown to be $\sim 100$ ps, which is much shorter than the SAW period of $860$ ps. Hence, it is promising to observe single-photon emission in the optimised device. In order to test single-photon emission, a Hanbury Brown-Twiss experimental setup has been employed to record an autocorrelation histogram of the SAW-driven EL signal at the single-electron regime. Suppression of autocorrelation coincidences at time delay $\Delta t = 0$ is evidence of photon antibunching. By fitting theoretical functions describing the SAW-driven EL signal, it is found that the second-order correlation function shows $g^{(2)}(0) = 0.39 \pm 0.05$, which is lower than the common criterion for a single-photon source $g^{(2)}(0) < 0.5$. Moreover, theoretical calculation and simulation suggest that, if a constant background signal can be filtered out, $\sim 80 \%$ of the SAW-driven EL is single-photon emission.

Les polyoxométallates (POMs) de type Keggin [XM12O40]n- (avec X=P… et M= W, Mo …) sont des oxydes moléculaires de métaux de transition à haut degré d’oxydation qui présentent des réductions successives et réversibles dans une gamme étroite de potentiel. Leur incorporation dans des dispositifs de mémoire moléculaire semble alors prometteuse. Nous avons développé leur intégration comme briques élémentaires via une approche “bottom-up” qui surmonte les limites de l’approche “top-down” plus commune. Cela nécessite un contrôle fin de leur greffage et de leur densité surfacique pour l’obtention de dispositifs performants. Nous avons donc développé le greffage covalent de POMs sur des surfaces à base de silicium et d’or. Des hybrides de POMs à terminaison diazonium BA3[PM11O39{SnC6H4C≡CC6H4N2}] (avec M=W or Mo) ont formé des monocouches complètes greffées sur des surfaces de silicium hydrogénées qui mettent en évidence l’influence du métal constitutif du fragment polyoxométallate sur les propriétés de transport de charges de la jonction. Le greffage d’hybrides de POM à terminaison acide carboxylique sur des substrats d’oxyde de silicium a aussi été développée. Un hybride de POM à terminaison aniline TBA4[PW11O39{SnC6H4C≡CC6H4NH2}] a été greffé en deux étapes via un couplage peptidique sur une monocouche à terminaison acide carboxylique sur surface d’or. Des monocouches compactes ont été obtenues mais pas de façon totalement reproductible et les premiers essais de dilution sont encourageants. Une nouvelle famille d’hybrides de POMs mixtes a été synthétisée : TBA4[PMoxW11-xO39{SnR}]. Cela permettra de combiner les propriétés redox du molybdène et la robustesse due au tungstène. / Keggin type polyoxometalates (POMs) [XM12O40]n- (with X=P… and M= W, Mo…) are molecular oxides of early transition metals with a high oxidation state. They present electrochemical successive reversible reduction waves in a narrow range of potential. This makes them good candidates to be incorporated into molecular memory devices. We chose a “bottom-up” approach where the POMs are the building blocks to overcome the limitation of the “top-down” process commonly used. A fine control of the POM grafting and of the surface density is essential to get better erase/writing time of the device. So, we developed POM hybrids for grafting them covalently onto silicon based and gold substrates. Diazonium-terminated POM hybrids (with M=W or Mo) lead to compact homogeneous monolayers onto hydrogenated silicon surfaces. Electrical measurements of the two analogous modified surfaces show different behaviour highlighting the role of the constituting POM metal into the charge transport. Carboxylic-terminated POM hybrids have also been grafted in a one-step process onto silicon oxide surface resulting in a smooth and dense monolayer. Then, an aniline-terminated POM hybrid has been grafted onto a carboxylic-terminated SAM of thiols onto gold thanks to a peptide coupling. Compact monolayers have been obtained without complete reproducibility and the first attempts of dilution are encouraging. A new family of POM hybrids have been synthesized: mixed-metal POM hybrids TBA4[PMoxW11-xO39{SnR}]. This will permit to combine the electrochemical properties of molybdenum and the robustness of tungsten.