Sudden cardiac death following cardiac arrest is a major killer in the industrialised world. The leading cause of sudden cardiac death are disturbances in the normal electrical activation of cardiac tissue, known as cardiac arrhythmia, which severely compromise the ability of the heart to fulfill the body's demand of oxygen. Ventricular fibrillation (VF) is the most deadly form of cardiac arrhythmia. Furthermore, electrical defibrillation through the application of strong electric shocks to the heart is the only effective therapy against VF. Over the past decades, a large body of research has dealt with the study of the mechanisms underpinning the success or failure of defibrillation shocks. The main mechanism of shock failure involves shocks terminating VF but leaving the appropriate electrical substrate for new VF episodes to rapidly follow (i.e. shock-induced arrhythmogenesis). A large number of models have been developed for the in silico study of shock-induced arrhythmogenesis, ranging from single cell models to three-dimensional ventricular models of small mammalian species. However, no extrapolation of the results obtained in the aforementioned studies has been done in human models of ventricular electrophysiology. The main reason is the large computational requirements associated with the solution of the bidomain equations of cardiac electrophysiology over large anatomically-accurate geometrical models including representation of fibre orientation and transmembrane kinetics. In this Thesis we develop simulation technology for the study of cardiac defibrillation in the human heart in the framework of the open source simulation environment Chaste. The advances include the development of novel computational and numerical techniques for the solution of the bidomain equations in large-scale high performance computing resources. More specifically, we have considered the implementation of effective domain decomposition, the development of new numerical techniques for the reduction of communication in Chaste's finite element method (FEM) solver, and the development of mesh-independent preconditioners for the solution of the linear system arising from the FEM discretisation of the bidomain equations. The developments presented in this Thesis have brought Chaste to the level of performance and functionality required to perform bidomain simulations with large three-dimensional cardiac geometries made of tens of millions of nodes and including accurate representation of fibre orientation and membrane kinetics. This advances have enabled the in silico study of shock-induced arrhythmogenesis for the first time in the human heart, therefore bridging an important gap in the field of cardiac defibrillation research.

L'équation intégrale du champ électrique (EFIE) est très utilisée pour résoudre des problèmes de diffusion d'ondes électromagnétiques grâce à la méthode aux éléments de frontière (BEM). En domaine fréquentiel, les systèmes matriciels émergeant de la BEM souffrent, entre autres, de deux problèmes de mauvais conditionnement : l'augmentation du nombre d'inconnues et la diminution de la fréquence entrainent l'accroissement du nombre de conditionnement. En conséquence, les solveurs itératifs requièrent plus d'itérations pour converger vers la solution, voire ne convergent pas du tout. En domaine temporel, ces problèmes sont également présents, en plus de l'instabilité DC qui entraine une solution erronée en fin de simulation. La discrétisation en temps est obtenue grâce à une quadrature de convolution basée sur les méthodes de Runge-Kutta implicites.Dans cette thèse, diverses formulations d'équations intégrales utilisant notamment des conditions d'impédance aux frontières (IBC) sont étudiées et préconditionnées. Dans une première partie en domaine fréquentiel, l'IBC-EFIE est stabilisée pour les basses fréquences et les maillages denses grâce aux projecteurs quasi-Helmholtz et à un préconditionnement de type Calderón. Puis une nouvelle forme d'IBC est introduite, ce qui permet la construction d'un préconditionneur multiplicatif. Dans la seconde partie en domaine temporel, l'EFIE est d'abord régularisée pour le cas d'un conducteur électrique parfait (PEC), la rendant stable pour les pas de temps larges et immunisée à l'instabilité DC. Enfin, unerésolution efficace de l'IBC-EFIE est recherchée, avant de stabiliser l'équation pour les pas de temps larges et les maillages denses. / The Electric Field Integral Equation (EFIE) is widely used to solve wave scattering problems in electromagnetics using the Boundary Element Method (BEM). In frequency domain, the linear systems stemming from the BEM suffer, amongst others, from two ill-conditioning problems: the low frequency breakdown and the dense mesh breakdown. Consequently, the iterative solvers require more iterations to converge to the solution, or they do not converge at all in the worst cases. These breakdowns are also present in time domain, in addition to the DC instability which causes the solution to be completely wrong in the late time steps of the simulations. The time discretization is achieved using a convolution quadrature based on Implicit Runge-Kutta (IRK) methods, which yields a system that is solved by Marching-On-in-Time (MOT). In this thesis, several integral equations formulations, involving Impedance Boundary Conditions (IBC) for most of them, are derived and subsequently preconditioned. In a first part dedicated to the frequency domain, the IBC-EFIE is stabilized for the low frequency and dense meshes by leveraging the quasi-Helmholtz projectors and a Calderón-like preconditioning. Then, a new IBC is introduced to enable the development of a multiplicative preconditioner for the new IBC-EFIE. In the second part on time domain,the EFIE is regularized for the Perfect Electric Conductor (PEC) case, to make it stable in the large time step regime and immune to the DC instability. Finally, the solution of the time domain IBC-EFIE is investigated by developing an efficient solution scheme and by stabilizing the equation for large time steps and dense meshes.

Cette thèse est dédiée à la résolution numérique tridimensionnelle des équations de Maxwell harmoniques, par des méthodes de décomposition de domaine couplant des résolutions par équations intégrales entre elles. Pour traiter les problèmes de diffraction d'ondes, la méthode des équations intégrales est un outil précieux. Elle consiste à paramétrer le champ électromagnétique solution par une source définie sur la surface de l'objet diffractant, solution d'une nouvelle équation linéaire (l'équation intégrale). Pour des applications à haute fréquence, le grand nombre d'inconnues (de l'ordre du million) nous oblige à utiliser un solveur itératif pour résoudre l'équation intégrale. Le problème du conditionnement des systèmes linéaires est alors crucial. De récents développements ont permis de construire une équation intégrale performante (la GCSIE) et de conditionnement stable avec la montée en fréquence. Cependant, la présence d'une cavité large et résonnante dans l'objet diffractant (telle que la cavité moteur d'un avion) dégrade le conditionnement de cette équation. Nous proposons deux méthodes de décomposition de domaine (DDM) afin de découpler le problème de la cavité du problème extérieur. La première (DDM en Y) s'exprime en fonction des opérateurs Dirichlet-to-Neumann Y, qui sont synthétisés via la résolution de problèmes métalliques par équations intégrales dans chaque sous-domaine. La seconde (DDM en S) s'exprime en fonction des opérateurs de scattering S, synthétisés par résolution de problèmes de type métal-impédant, donc bien posés à toute fréquence. La DDM en S permet ainsi de se débarrasser des phénomènes de résonance dans les cavités. Nous proposons dans un premier temps un préconditionneur analytique pour la DDM en Y, basé sur l'opérateur électromagnétique de simple couche. Nous calculons ensuite les modes guidés le long d'un cylindre infini tangent à la cavité près de l'interface, et nous diagonalisons les opérateurs Dirichlet-to-Neumann et scattering dans la base des traces de modes guidés sur l'interface. On extrait de cette étude deux préconditionneurs spectraux respectivement pour la DDM en Y et la DDM en S. Les résultats numériques confirment l'efficacité des préconditionneurs proposés / This work is dedicated to the numerical solution of the tridimensional harmonic Maxwell equations, using domain decomposition methods coupling integral equations between them. To deal with scattering problems, integral equations methods are a precious tool. They allow to look for the electromagnetic field by parameterizing it with a source only defined on the boundary of the scattering object, solution of a new linear equation (the integral equation). For applications at high frequency, the great number of unknowns forces the use of iterative methods. To accelerate the solution of integral equations, one moreover has to ensure the good condition number of the linear systems, or to propose well-suited preconditioners. An efficient method, the GCSIE, was developed in Onera. It is an intrinsically well-conditioned integral equation whose condition number remains stable whith the frequency increase. However, the existence of large and resonant cavities (such as air intakes) deteriorates the condition number. In order to circumvent this problem, we propose two domain decomposition methods (DDM) allowing to decouple the exterior problem from the problem of the cavities. The first one (Y-DDM) is based on Dirichlet-to-Neumann operators Y, which are built through the solution of metallic problems using integral equations in each subdomain. The second one (S-DDM) is based on scattering operators S, built through the solution of problems of metallic-impedant type, which are well-posed at any frequency. The S-DDM allows to avoid the resonance phenomena inside the cavities. First, we propose an analytic preconditioner for the Y-DDM, based on the electromagnetic single layer operator. We then calculate the modes guided along an artificial infinite cylinder, that is tangent to the cavity near the interface. We diagonalize the Dirichlet-to-Neumann and scattering operators in the basis of the traces of the guided modes on the interface. We deduce from this study two spectral preconditioners for the Y-DDM and the S-DDM. The numerical results confirm the efficiency of the employed preconditioners.

INTRODUÇÃO: O transplante hepático se consolidou como terapêutica para crianças com doença hepática em estágio terminal. A principal indicação nessa faixa etária é a atresia das vias biliares, doença colestática que leva à cirrose hepática na maioria dos casos nos primeiros anos de vida mesmo nas crianças submetidas a portoenterostomia. Essas crianças evoluem entre outras complicações com desnutrição grave e necessitam do transplante precocemente. A relação ideal do peso do enxerto hepático e o peso do receptor varia de 1 a 3%. No entanto, quando o transplante é realizado em crianças em idade inferior a 2 anos, essa relação na maioria das vezes é maior do que 5%. Essa situação é conhecida como large-for-size, em que a perfusão sanguínea do enxerto pode ser insuficiente, o que leva a disfunção do órgão. Os mecanismos de lesão neste cenário ainda não estão bem esclarecidos. A lesão de isquemia-reperfusão (IR) é apontada como um dos fatores para o mal funcionamento do enxerto bem como de outras complicações. Nessa lesão estão envolvidos elementos da cascata inflamatória culminando em morte celular por apoptose bem como mecanismos de regeneração celular. Um fenômeno conhecido que atenua a lesão de IR é o pré-condicionamento isquêmico (PCI). Nenhum trabalho experimental foi realizado com o objetivo de avaliar a lesão hepática na situação large-for-size e o efeito do PCI nessa situação. MÉTODOS: Para a padronização do modelo foram realizados 8 transplantes, tendo sido testados os pesos dos animais, relação entre o peso do enxerto hepático e peso do receptor e técnica cirúrgica. Na fase experimental, foram realizados 21 transplantes e os animais foram divididos em 3 grupos. No primeiro grupo (controle) o transplante era realizado com doadores e receptores com pesos similares. Para caracterização da situação large-for-size, no segundo grupo os doadores tinham aproximadamente o dobro do peso dos receptores (LFS). No terceiro grupo a distribuição de peso foi semelhante a do segundo, porém era realizado o PCI no doador (LFS+PCI). Os procedimentos foram realizados sob anestesia geral e técnica asséptica e os animais foram mantidos vivos por 3 horas após a reperfusão. Foram obtidas amostras de sangue em 3 períodos (logo após abertura da cavidade abdominal, 1 e 3 horas após reperfusão) e fragmentos de fígado 1 e 3 horas após reperfusão. As amostras de sangue foram submetidas a gasometria arterial, dosagem de sódio, potássio e enzimas hepatocelulares. Os fragmentos de tecido hepático foram submetidos a análise histológica pela coloração da hematoxilina-eosina (HE) e a análise de quantificação da expressão dos genes pró-apoptótico (Bax), anti-apoptótico (Bcl- XL), da óxido nítrico-sintase endotelial (eNOS) e da interleucina-6 (IL-6). RESULTADOS: A mortalidade observada foi de 28%. A relação peso do enxerto e peso do paciente foi de 2,9% no grupo controle, 6,3% no grupo LFS e 6,4% no grupo LFS+PCI. A natremia 1 hora após a reperfusão foi mais elevada no grupo controle do que nos demais, já com relação ao potássio, a dosagem foi mais baixa nesse grupo no mesmo intervalo. A dosagem da aspartato aminotransferase (AST) foi mais elevada nos grupos LFS e LFS+PCI do que no controle. A análise histológica não Resumo demonstrou diferença entre os grupos. A expressão do gene Bax foi mais elevada no grupo LFS do que nos demais, enquanto que a do gene eNOS foi mais acentuada no grupo LFS+PCI do que nos demais nos dois intervalos estudados. Após 3 horas de reperfusão a expressão do gene IL-6 foi maior no grupo LFS+PCI. CONCLUSÕES: O modelo de transplante hepático large-for-size em suínos foi exequível e adequado para a pesquisa. A lesão IR foi mais acentuada no grupo LFS do que no grupo controle e apesar de não ter alterado a elevação de enzimas hepatocelulares, o PCI teve papel de aumentar a expressão dos genes IL-6 e eNOS e diminuir a do gene Bax / INTRODUCTION: Liver transplantation has been established as therapy for children with end-stage liver disease. The main indication is biliary atresia, cholestatic disease that leads to cirrhosis in most cases even in children undergoing portoenterostomy. These children develop other complications as severely malnutrition and need early transplantation. The ideal graft to body weight ratio (GBWR) for transplantation varies from 1 to 3%. However, when transplantation is performed in children younger than 2 years, this ratio in most cases is higher than 5%. This situation, known as large-for-size, may be related to insufficient graft perfusion and liver disfunction. The mechanisms of injury involved in this scenario are not well established. Ischemia-reperfusion injury (IRI) has been related as a factor for poor graft function and other complications. Inflammatory cascade culminating in cell death by apoptosis and cellular mechanisms of regeneration are the elements involved in IRI. Ischemic preconditioning (IPC) is a phenomenon that attenuates IR injury. There are no experimental studies conducted to evaluate the hepatic injury in large-for-size situation and the effect of IPC in this situation. METHODS: 8 procedures were required for standardize the model. Weight of the animals, GBWR and surgical technique were evaluated. In the experimental phase, 21 transplantations were performed. The animals were divided in three groups. In the first group (control), liver transplantation was performed with donors and recipients with similar weights. To characterize large-for-size situation, donors of the second group had approximately twice the weight receptor (LFS). In the third group the weight distribution was similar to the second but IPC was performed in donor (LFS+IPC). Procedures were performed under general anesthesia and aseptic technique and the animals were kept alive for 3 hours after reperfusion. Blood samples were obtained at three times (immediately after opening the abdominal cavity, 1 and 3 hours after reperfusion) and fragments of liver 1 and 3 hours after reperfusion. Blood samples were analyzed for arterial blood gases, sodium, potassium and hepatocellular enzymes. The fragments of liver tissues were subjected to histological analysis by hematoxylin-eosin staining (HE). Molecular biology involved quantification of the expression of Bax, Bcl-XL, endothelial nitric oxide synthase (eNOS) and interleukin- 6 (IL-6) genes. RESULTS: The mortality rate was 28%. The GBWR was 2.9% in the control group, 6.3% in the LFS group and 6.4% in the LFS+ICP group. The natremia 1 hour after reperfusion was higher in the control group and potassium dosage was lower in this group in the same period. The dosage of aspartate aminotransferase (AST) was higher in LFS and LFS+IPC groups than control. Histological analysis showed no difference between groups. The Bax gene expression was higher in the LFS, while the eNOS gene was more expressed in LFS + PCI group. After 3 hours of reperfusion the expression of IL-6 gene was higher in the PCI + LFS. CONCLUSIONS: The model of large-for-size liver transplantation in swine was feasible and appropriate for the research. IR injury was more pronounced in the Summary group LFS than in the control group and despite not having changed the elevation of hepatocellular enzymes, the PCI was responsible for increasing the expression of IL- 6 and eNOS and decreasing the Bax gene expression

Le postconditionnement cardiaque est paradoxal chez le rat. Certains auteurs ont montré que cet animal pouvait être protégé par le postconditionnement alors que d’autres ont montré qu’il était inefficace. L’objectif de notre travail était d’éclaircir cette situation et d’établir un lien entre la régulation des fonctions mitochondriales et la réussite ou l’échec du postconditionnement. Nous avons montré sur un modèle in vivo que le rat est sensible au postconditionnement cardiaque sous certaines conditions. Il semble que la réussite de cette thérapie nécessite à la fois une préservation de la phosphorylation oxydative, une inhibition de l’ouverture du mPTP et une diminution de la production de ROS. Nous avons également montré que le complexe I de la chaîne respiratoire régule l’ouverture du mPTP en liaison avec l’état de la Cyp D. L’ensemble de nos résultats montrent que le rat n’est pas réfractaire au postconditionnement, mais pour que celui-ci soit efficace, il est nécessaire de préserver l’intégrité des différentes fonctions mitochondriales. La cardioprotection et la régulation des fonctions mitochondriales sont donc spécifiquement liées au modèle utilisé / Cardiac postconditioning is paradoxical in rat. Certain authors showed that this animal could be protected by postconditioning whereas others showed that it was ineffective. The objective of our work was to clarify this situation and to establish a link between the regulation of the mitochondrial functions and the success or the failure of postconditioning. We showed on an in vivo model that the rat is sensitive to cardiac postconditioning under certain conditions. It seems that the success of this therapy requires at the same time a safeguarding of oxidative phosphorylation, an inhibition of the mitochondrial permeability transition pore opening and a reduction in the ROS production. We also showed that complex I of the respiratory chain controls the opening of the mPTP in relation with the state of Cyp D. Our results show that the rat is not insensitive to postconditioning, but it is necessary to preserve the integrity of the mitochondrial functions to be effective. The cardioprotection and the regulation of the mitochondrial functions are specifically related to the model used

La thérapie cellulaire suscite de grands espoirs dans le domaine cardiovasculaire. Cependant les premières études humaines sont décevantes. Parmi les explications avancées, citons un mauvais choix de cellules, une méthode de délivrance inadéquate, une préparation des cellules et des tissus hôtes insuffisante ou une trop grande mortalité cellulaire après injection.Durant ce travail nous avons voulu explorer 3 pistes d’optimisation de la thérapie cellulaire à visée pro-angiogénique utilisant les cellules souches mésenchymateuses (MSC) et contribué à l’exploration des mécanismes mis en jeu, en particulier en explorant le rôle du système Wnt/Frizzled.Nous avons tout d’abord étudié un système original de délivrance de cellules utilisant un « patch » musculaire cousu en regard d’un myocarde infarci de souris. Puis nous avons étudié l’effet d’une surexpression de sFRP1, inhibiteur de la voie Wnt, sur un modèle de matrice sous cutanée. Enfin, nous avons testé l’hypothèse qu’un préconditionnement hypoxique des cellules permettrait une meilleure survie cellulaire et améliorerait la réparation vasculaire et tissulaire après ischémie de patte chez la souris.Nos résultats permettent notamment de montrer que les MSC ont des capacités d’invasion des tissus ischémiques et qu’elles se différencient en péricytes en formant un réseau tridimensionnel de soutien aux cellules endothéliales. Par ailleurs, via sFRP1, nous mettons en évidence un rôle du système Wnt/Fzd dans l’effet pro-angiogénique. Enfin, nous montrons l’intérêt du préconditionnement hypoxique dont les effets sont médiés par Wnt4.L’ensemble de ces données permet d’envisager des voies d’optimisation de la thérapie cellulaire. / Some of the challenges facing stem-cell therapy for cardiac disease are which type of stem cell or progenitor cell is the best candidate for therapy, how to survive in the low oxygen environment of ischemic myocardium. Here we studied the potential of mesenchymal stem cells (MSCs) as vascular progenitor cells in vitro and in vivo, and we studied the effects of sFRP-1/Wnt signaling modulation or hypoxia on MSC properties. First, we demonstrated the beneficial effect of MSC application on ischemic heart repair using an original surgical model (patch) to deliver stem cells. This study showed that the contribution of the MSCs in the mouse infarcted myocardium was beneficial either on the scar (increase in angiogenesis, in cell proliferation, reduction in ventricular remodeling) or on the trophicity of the patch.Then we characterized the angiogenic properties of MSC in vitro and in vivo. Our data demonstrate that MSCs could be recruited and formed vascular structures around endothelial tubes. We showed in vivo that the surexpression of sFRP-1 (regulating factor of the Wnt system) in MSCs increased their potential of pericyte-like cells correlated with an increased maturation of the vessels via an intracellular GSK-3 dependent pathway in MSCs. Our next objective was to investigate the effects of hypoxia exposure on MSC before implantation for vascular and tissue regeneration in mice with hind limb ischemia. Our data suggest that hypoxic preconditioning has a critical role on MSC dynamic functions, shifting MSC location in situ to enhance ischemic tissue recovery, facilitating vascular cell mobilization and skeletal myoblast regeneration via a paracrine Wnt dependent mechanism.

Introdução: Acredita-se que o diabetes mellitus possa alterar mecanismos celulares do miocárdio tornando-o mais sensível a um insulto isquêmico, e que esta menor resistência do miocárdio isquêmico induzido pelo diabetes possa ser um dos motivos para o pior prognóstico observado em pacientes com doença arterial coronariana e diabetes. Um dos principais mecanismos adaptativos protetores do miocárdio é o precondicionamento isquêmico, sendo este desencadeado por curtos períodos de isquemia seguidos por reperfusão e que tornam o tecido mais resistente a um insulto isquêmico grave e prolongado. Em humanos, o precondicionamento isquêmico pode ser observado durante testes ergométricos sequenciais, nos quais a melhora em parâmetros isquêmicos no segundo teste ergométrico quando comparado ao primeiro é uma metodologia consagrada para o estudo clínico deste fenômeno. Estudos experimentais demonstram resultados controversos em relação à interferência do diabetes sobre o fenômeno do precondicionamento, e estudos com humanos são escassos e inconclusivos. Assim, ainda é incerto se o diabetes pode afetar a expressão do precondicionamento isquêmico em pacientes com doença arterial coronariana. Objetivos: Identificar se o diabetes mellitus interfere no fenômeno do precondicionamento isquêmico em pacientes com doença arterial coronariana. Métodos: Pacientes com doença arterial coronariana comprovada por cineangiocoronariografia diagnóstica, função ventricular sistólica preservada e com angina ou teste ergométrico positivo para isquemia miocárdica foram submetidos a dois testes ergométricos sequenciais com intervalo de 30 minutos. Parâmetros isquêmicos foram comparados entre pacientes com e sem diabetes mellitus. O precondicionamento isquêmico foi considerado presente quando o tempo para a depressão em 1,0 mm do segmento ST (T-1mm) foi maior no segundo teste sequencial comparado ao primeiro. Também se mensurou o duplo-produto (frequência cardíaca multiplicada pela pressão arterial sistólica) no momento do T- 1mm. Os testes foram analisados por dois cardiologistas experientes, independentes. Resultados: De 2.140 pacientes consecutivos com doença arterial coronariana, 361 apresentavam critérios para inclusão nesse estudo. Destes, 174 pacientes (64,2 ± 7,6 anos) foram submetidos aos testes ergométricos sequenciais para identificação e caracterização do precondicionamento isquêmico; 86 apresentavam diabetes mellitus (grupo 1) e 88 não apresentavam diabetes mellitus (grupo 2). Os dois grupos foram semelhantes em relação às principais características demográficas, com exceção de infarto do miocárdio prévio e perfil lipídico. No primeiro grupo, 62 pacientes (72,1%) manifestaram o precondicionamento isquêmico e no segundo, 60 (68,2%) manifestaram o precondicionamento isquêmico (P=0,62). Analisando-se os pacientes que expressaram o fenômeno, a melhora do T-1mm foi similar entre os dois grupos (média da melhora do tempo entre os testes 1 e 2: 79,4 ± 47,6 x 65,5 ± 36,4 segundos, respectivamente para os grupos 1 e 2, P=0,12). Em relação ao duploproduto no momento do T-1mm, os pacientes com diabetes apresentaram melhora expressiva em relação aos pacientes sem diabetes (média da melhora do duploproduto 3011 ± 2430 x 2081 ± 2139 bpm x mmHg, respectivamente para os grupos 1 e 2, P=0,01). A análise da melhora das arritmias e da morfologia da depressão do segmento ST nos testes ergométricos sequenciais não mostrou diferenças entre os dois grupos de pacientes. Conclusão: Neste estudo, o diabetes mellitus tipo 2 não impediu o surgimento do PI. Além disso, o diabetes esteve associado à melhora significativa do esforço cardíaco e do consumo miocárdico de oxigênio, caracterizados pelo duplo-produto . / Background: It\'s postulated that diabetes mellitus may impair myocardial cellular mechanisms turning it more sensitive to ischemic injuries, and that this lower resistance of the ischemic myocardium induced by diabetes may be one reason for the poor prognosis observed in patients with both coronary artery disease and diabetes. One major adaptive myocardial protective mechanism is ischemic preconditioning, which is triggered by brief ischemia followed by reperfusion that turns the myocardium more resistant to a prolonged ischemic insult. In humans, ischemic preconditioning can be observed during sequential exercise tests, in which the improvement in ischemic parameters in the second exercise test compared to the first is a methodology devoted to the clinical study of the phenomenon. However, experimental studies have shown conflicting results about the interference of diabetes on ischemic preconditioning, and the few human studies are scarce and inconclusive. Thus, it\'s still uncertain whether diabetes may affect ischemic preconditioning in coronary artery disease patients. Objectives: Identify whether type 2 diabetes mellitus intervenes on myocardial ischemic preconditioning in symptomatic coronary artery disease patients. Methods: Symptomatic multivessel coronary artery disease patients with preserved systolic ventricular function and a positive exercise test underwent two sequential exercise tests to demonstrate ischemic preconditioning. Tests were performed with a 30 minutes interval between them. Ischemic parameters were compared among patients with and without type 2 diabetes mellitus. Ischemic preconditioning was considered present when the time to 1.0 mm ST deviation (T-1mm) was greater in the second of 2 exercise tests. Rate pressure-product (heart rate multiplied by systolic arterial pressure) at T-1mm was also assessed. Sequential exercise tests were analyzed by 2 independent cardiologists. Results: Of the 2,140 consecutive coronary artery disease patients screened, 361 met inclusion criteria, and 174 patients (64.2 ± 7.6 years) completed the study protocol. Of these, 86 had the diagnosis of type 2 diabetes. The 2 groups were similar regarding the main demographic characteristics, except for the rate of previous myocardial infarction and lipid profile. Among diabetic patients, 62 (72.1%) manifested an improvement in ischemic parameters consistent with ischemic preconditioning, whereas among nondiabetic patients, 60 (68.2%) manifested ischemic preconditioning (P=0.62). The analysis of patients who demonstrated ischemic preconditioning showed similar improvement in the time to 1.0 mm ST deviation between diabetic and nondiabetic groups (79.4 ± 47.6 vs 65.5 ± 36.4 sec, respectively, P=0.12). Regarding rate pressure-product, the improvement was greater in diabetic compared to nondiabetic patients (3011 ± 2430 vs 2081 ± 2139 bpm x mmHg, respectively, P=0.01). The analysis of arrhythmias and ST-segment deviation morphology during sequential exercise tests did not show differences between the 2 groups of patients. Conclusion: In this study, type 2 diabetes mellitus did not prevent the occurrence of ischemic preconditioning. In addition, diabetes was associated with significant improvement in cardiac stress and myocardial oxygen consumption, characterized by rate pressure-product

INTRODUÇÃO: O transplante hepático se consolidou como terapêutica para crianças com doença hepática em estágio terminal. A principal indicação nessa faixa etária é a atresia das vias biliares, doença colestática que leva à cirrose hepática na maioria dos casos nos primeiros anos de vida mesmo nas crianças submetidas a portoenterostomia. Essas crianças evoluem entre outras complicações com desnutrição grave e necessitam do transplante precocemente. A relação ideal do peso do enxerto hepático e o peso do receptor varia de 1 a 3%. No entanto, quando o transplante é realizado em crianças em idade inferior a 2 anos, essa relação na maioria das vezes é maior do que 5%. Essa situação é conhecida como large-for-size, em que a perfusão sanguínea do enxerto pode ser insuficiente, o que leva a disfunção do órgão. Os mecanismos de lesão neste cenário ainda não estão bem esclarecidos. A lesão de isquemia-reperfusão (IR) é apontada como um dos fatores para o mal funcionamento do enxerto bem como de outras complicações. Nessa lesão estão envolvidos elementos da cascata inflamatória culminando em morte celular por apoptose bem como mecanismos de regeneração celular. Um fenômeno conhecido que atenua a lesão de IR é o pré-condicionamento isquêmico (PCI). Nenhum trabalho experimental foi realizado com o objetivo de avaliar a lesão hepática na situação large-for-size e o efeito do PCI nessa situação. MÉTODOS: Para a padronização do modelo foram realizados 8 transplantes, tendo sido testados os pesos dos animais, relação entre o peso do enxerto hepático e peso do receptor e técnica cirúrgica. Na fase experimental, foram realizados 21 transplantes e os animais foram divididos em 3 grupos. No primeiro grupo (controle) o transplante era realizado com doadores e receptores com pesos similares. Para caracterização da situação large-for-size, no segundo grupo os doadores tinham aproximadamente o dobro do peso dos receptores (LFS). No terceiro grupo a distribuição de peso foi semelhante a do segundo, porém era realizado o PCI no doador (LFS+PCI). Os procedimentos foram realizados sob anestesia geral e técnica asséptica e os animais foram mantidos vivos por 3 horas após a reperfusão. Foram obtidas amostras de sangue em 3 períodos (logo após abertura da cavidade abdominal, 1 e 3 horas após reperfusão) e fragmentos de fígado 1 e 3 horas após reperfusão. As amostras de sangue foram submetidas a gasometria arterial, dosagem de sódio, potássio e enzimas hepatocelulares. Os fragmentos de tecido hepático foram submetidos a análise histológica pela coloração da hematoxilina-eosina (HE) e a análise de quantificação da expressão dos genes pró-apoptótico (Bax), anti-apoptótico (Bcl- XL), da óxido nítrico-sintase endotelial (eNOS) e da interleucina-6 (IL-6). RESULTADOS: A mortalidade observada foi de 28%. A relação peso do enxerto e peso do paciente foi de 2,9% no grupo controle, 6,3% no grupo LFS e 6,4% no grupo LFS+PCI. A natremia 1 hora após a reperfusão foi mais elevada no grupo controle do que nos demais, já com relação ao potássio, a dosagem foi mais baixa nesse grupo no mesmo intervalo. A dosagem da aspartato aminotransferase (AST) foi mais elevada nos grupos LFS e LFS+PCI do que no controle. A análise histológica não Resumo demonstrou diferença entre os grupos. A expressão do gene Bax foi mais elevada no grupo LFS do que nos demais, enquanto que a do gene eNOS foi mais acentuada no grupo LFS+PCI do que nos demais nos dois intervalos estudados. Após 3 horas de reperfusão a expressão do gene IL-6 foi maior no grupo LFS+PCI. CONCLUSÕES: O modelo de transplante hepático large-for-size em suínos foi exequível e adequado para a pesquisa. A lesão IR foi mais acentuada no grupo LFS do que no grupo controle e apesar de não ter alterado a elevação de enzimas hepatocelulares, o PCI teve papel de aumentar a expressão dos genes IL-6 e eNOS e diminuir a do gene Bax / INTRODUCTION: Liver transplantation has been established as therapy for children with end-stage liver disease. The main indication is biliary atresia, cholestatic disease that leads to cirrhosis in most cases even in children undergoing portoenterostomy. These children develop other complications as severely malnutrition and need early transplantation. The ideal graft to body weight ratio (GBWR) for transplantation varies from 1 to 3%. However, when transplantation is performed in children younger than 2 years, this ratio in most cases is higher than 5%. This situation, known as large-for-size, may be related to insufficient graft perfusion and liver disfunction. The mechanisms of injury involved in this scenario are not well established. Ischemia-reperfusion injury (IRI) has been related as a factor for poor graft function and other complications. Inflammatory cascade culminating in cell death by apoptosis and cellular mechanisms of regeneration are the elements involved in IRI. Ischemic preconditioning (IPC) is a phenomenon that attenuates IR injury. There are no experimental studies conducted to evaluate the hepatic injury in large-for-size situation and the effect of IPC in this situation. METHODS: 8 procedures were required for standardize the model. Weight of the animals, GBWR and surgical technique were evaluated. In the experimental phase, 21 transplantations were performed. The animals were divided in three groups. In the first group (control), liver transplantation was performed with donors and recipients with similar weights. To characterize large-for-size situation, donors of the second group had approximately twice the weight receptor (LFS). In the third group the weight distribution was similar to the second but IPC was performed in donor (LFS+IPC). Procedures were performed under general anesthesia and aseptic technique and the animals were kept alive for 3 hours after reperfusion. Blood samples were obtained at three times (immediately after opening the abdominal cavity, 1 and 3 hours after reperfusion) and fragments of liver 1 and 3 hours after reperfusion. Blood samples were analyzed for arterial blood gases, sodium, potassium and hepatocellular enzymes. The fragments of liver tissues were subjected to histological analysis by hematoxylin-eosin staining (HE). Molecular biology involved quantification of the expression of Bax, Bcl-XL, endothelial nitric oxide synthase (eNOS) and interleukin- 6 (IL-6) genes. RESULTS: The mortality rate was 28%. The GBWR was 2.9% in the control group, 6.3% in the LFS group and 6.4% in the LFS+ICP group. The natremia 1 hour after reperfusion was higher in the control group and potassium dosage was lower in this group in the same period. The dosage of aspartate aminotransferase (AST) was higher in LFS and LFS+IPC groups than control. Histological analysis showed no difference between groups. The Bax gene expression was higher in the LFS, while the eNOS gene was more expressed in LFS + PCI group. After 3 hours of reperfusion the expression of IL-6 gene was higher in the PCI + LFS. CONCLUSIONS: The model of large-for-size liver transplantation in swine was feasible and appropriate for the research. IR injury was more pronounced in the Summary group LFS than in the control group and despite not having changed the elevation of hepatocellular enzymes, the PCI was responsible for increasing the expression of IL- 6 and eNOS and decreasing the Bax gene expression

Extraction of physiological and clinical information hidden in biosignals, such as cardiac and neural signals, is an important and fascinating field of research. Noninvasive assessment of the physiological parameters of a patient enables to study the physiology and pathophysiology of the investigated system, with minimal interference and inconvenience. This approach may also help to assess noninvasively the clinical condition of the patient. The primary focus of this study is therefore to extend the arsenal of research tools for the noninvasive investigation of the neural and cardiac systems. The approaches developed in this work concern two major directions: The first direction relies on the analysis of cardiac and neural responses during hypoxia. Hypoxia-ischemia remains a great challenge to the researchers, since it triggers complex responses at different levels in the organism. The functional recovery depends on a number of factors among which the state of autonomic nervous system (ANS) regulation plays an important role. Two different applications were considered in this framework. The first application studied the effect of global ischemic preconditioning on the heart rate variability (HRV) response to the asphyxia insult. Using linear (time and frequency domain) and nonlinear (approximate entropy and parameters of Poincare plots) measures, we evaluated the dynamic time course of the HRV response to the asphyxia insult and the effect of preconditioning on the autonomic neurocardiac control. Our results show for the first time that global ischemic preconditioning influences the HRV response to the asphyxia injury. The neuroprotective effect of preconditioning translates into a faster recovery of the basal HRV and the autonomic modulation of the heart. For the preconditioned group, at about 90 min after the asphyxic insult, the autonomic neural balance (measured by LF/HF ratio) appears fully recovered. Another application addressed the problem of phase synchronization analysis of EEG signals during monitoring of recovery process following brain injury episode. The concept of phase synchronization offers a new perspective on the understanding and quantification of the dynamical interactions established among coupled systems. In this thesis, we present a new approach for the identification of the degree of interaction between two complex dynamical systems from experimental data analysis. We use the empirical-mode-decomposition (EMD) technique to decompose the output signals into a number of elementary orthogonal modes with well defined instantaneous attributes (IMFs). The second direction addressed the problem of correlations between anticipatory pursuit eye movements and the neural response in the Supplementary Eye Fields (SEF) of the Macaque monkey. Anticipatory pursuit is a smooth movement of the eye occurring before the appearance of an expected moving target. The expectation of the subject is based on a subjective estimation of the probability that the target will move in a given direction. Recently, it has been suggested that the SEF could play a role in using past experience to guide anticipatory pursuit. This hypothesis is currently being tested at the single neuron level. In the behaving monkey, it has been shown that electrical microstimulation in the SEF can facilitate smooth pursuit initiation towards a moving target, suggesting that activation of the SEF might change the internal gain of the smooth pursuit pathway. In this study, we favored anticipatory responses in monkeys by using a cognitive cue, which produces a different anticipatory pursuit response than the one observed in previous studies, based on repetition. / H εξαγωγή φυσιολογικών και κλινικών πληροφοριών οι οποίες είναι κρυμμένες σε βιοσήματα όπως τα καρδιακά και νευροφυσιολογικά σήματα είναι ένας σημαντικός και πολύ ενδιαφέρον τομέας έρευνας. Μη επεμβατική αξιολόγηση των φυσιολογικών παραμέτρων ενός ασθενή επιτρέπει την μελέτη της φυσιολογίας και παθολογίας του μελετούμενου συστήματος με τις λιγότερες παρεμβολές και ενόχληση. Η προσέγγιση αυτή μπορεί επίσης να βοηθήσει στην μη επεμβατική αξιολόγηση της κλινικής κατάστασης του ασθενή. Η πρώτη προσέγγιση της μελέτης αυτής είναι να επεκτείνει το οπλοστάσιο των ερευνητικών εργαλείων για την μη επεμβατική αναζήτηση του νευρικού και καρδιακού συστήματος. Οι προσεγγίσεις που αναπτύσσονται σε αυτή τη δουλειά αφορούν δύο κύριες κατευθύνσεις: Η πρώτη κατεύθυνση υπόκειται στην ανάλυση των καρδιακών και νευροφυσιολογικών αποκρίσεων κατά τη διάρκεια της υποξίας. Η ισχεμία – υποξία παραμένει μια μεγάλη πρόκληση στους ερευνητές εφόσον πυροδοτεί πολύπλοκες αποκρίσεις σε διαφορετικά επίπεδα στον οργανισμό. Η λειτουργική αποκατάσταση εξαρτάται από έναν αριθμό συντελεστών μεταξύ των οποίων ο έλεγχος της κατάστασης του αυτόνομου νευρικού συστήματος παίζει έναν πολύ σημαντικό ρόλο. Δύο διαφορετικές εφαρμογές ελήφθησαν υπόψη στο πλαίσιο αυτό. Η πρώτη εφαρμογή μελέτησε το φαινόμενο της ολικής ισχαιμικής προκατάστασης στην μεταβλητότητα του καρδιακού ρυθμού (heart rate variability - HRV) σε προσβολή από ασφυξία. Χρησιμοποιώντας γραμμικές (στον τομέα του χρόνου και των συχνοτήτων) και μη γραμμικές (υπολογισμός εντροπίας και παραμέτρων των γραφημάτων Poincare) τεχνικές υπολογίσαμε την δυναμική χρονική εξέλιξη της HRV απόκρισης στην προσβολή από ασφυξία και η επίπτωση της προ-κατάστασης στο αυτόνομο νευροκαρδιολογικό έλεγχο. Τα αποτελέσματά μας έδειξαν για πρώτη φορά ότι η ολική ισχαιμική προκατάσταση επηρεάζει την HRV απόκριση στον τραυματισμό από την ασφυξία. Η νευροπροστατευτική επίπτωση της προκατάστασης μεταφράζεται σε μία γρηγορότερη αποκατάσταση του βασικού HRV και μία αυτόνομη εναρμόνιση της καρδιάς. Για την ομάδα με την προκατάσταση σε περίπου 90 λεπτά μετά την προσβολή από ασφυξία, η αυτόνομη νευρολογική ισορροπία (μετρούμενη από τον λόγο χαμηλών προς υψηλών συχνοτήτων εμφανίζεται πλήρως αποκαταστημένη. Μία άλλη εφαρμογή απευθύνεται στο πρόβλημα της ανάλυσης του συγχρονισμού φάσεων των σημάτων Ηλεκτροεγκεφαλογραφήματος κατά τη διάρκεια παρακολούθησης της διαδικασίας αποκατάστασης μετά από επεισόδιο εγκεφαλικής βλάβης. Η ιδέα του συγχρονισμού φάσεων προσφέρει μία νέα προοπτική στην κατανόηση και ποσοτικοποίηση των δυναμικών αλληλεπιδράσεων μεταξύ συστημάτων συζευγμένων ταλαντωτών. Σε αυτή τη διδακτορική διατριβή παρουσιάζουμε μια νέα προσέγγιση για την ανίχνευση του βαθμού της αλληλεπίδρασης μεταξύ δύο πολύπλοκων δυναμικών συστημάτων από την ανάλυση πειραματικών δεδομένων. Χρησιμοποιούμε την τεχνική του εμπειρικού τρόπου αποδόμησης (empirical-mode-decomposition EMD) για να διασπάσουμε τα σήματα εξόδου σε έναν αριθμό βασικών ορθογώνιων μερών με πολύ καλά καθορισμένες στιγμιαίες ιδιότητες (instantaneous attributes IMFs). Η δεύτερη κατεύθυνση είναι το πρόβλημα των συσχετίσεων μεταξύ προνοητικών κινήσεων των ματιών και των νευροφυσιολογικών αποκρίσεων στα παιδία των ματιών (Supplementary Eye Fields SEF) πιθήκων Macaque. Οι προνοητικές κινήσεις είναι απαλές κινήσεις των ματιών που συμβαίνουν πριν την εμφάνιση ενός αναμενόμενου κινούμενου στόχου. Η αναμονή από το υποκείμενο βασίζεται σε έναν υποκειμενικό υπολογισμό της πιθανότητας ότι ο στόχος θα κινηθεί σε μια δεδομένη κατεύθυνση. Πρόσφατα, έχει υποτεθεί ότι τα SEF μπορούν να παίζουν ρόλο στην χρησιμοποίηση παλαιών εμπειριών στην καθοδήγηση αναμενόμενων κινήσεων. Αυτή η υπόθεση έχει ελεγχθεί στο επίπεδο ενός μόνο νευρώνα. Στον πίθηκο έχει βρεθεί ότι ο ηλεκτρικός μικροερεθισμός στο SEF μπορεί να διευκολύνει την ομαλή έναρξη της κίνησης προς έναν κινούμενο στόχο, συνιστώντας ότι η ενεργοποίηση του SEF μπορεί να αλλάξει την εσωτερική απόδοση του δικτύου της ομαλής κίνησης. Σε αυτή τη μελέτη, ενισχύσαμε την εκκίνηση των ομαλών κινήσεων των πιθήκων προς ένα κινούμενο στόχο η οποία παράγει μία διαφορετική προνοητική κίνηση αυτής που παρατηρείται σε προηγούμενες μελέτες η οποία βασίζεται σε επανάληψη.

Η χρήση επαναληπτικών τεχνικών προσαρμοστικής ισοστάθμισης διαύλου αποτελεί μια σχετικά πρόσφατη και πολλά υποσχόμενη μέθοδο αντιμετώπισης του φαινομένου της διασυμβολικής παρεμβολής που εισάγεται από το κανάλι λόγω του φαινομένου της πολυδιόδευσης. Ο αλγόριθμος που έχει επικρατήσει στις περισσότερες προσαρμοστικές εφαρμογές είναι ο ελαχίστων μέσων τετραγώνων (LMS). Διακρίνεται για την απλότητά του, έχει όμως φτωχές ιδιότητες σύγκλισης. Η μέθοδος των αναδρομικών ελαχίστων τετραγώνων (RLS) είναι επίσης αρκετά διαδεδομένη και κατέχει υπερέχουσες ιδιότητες σύγκλισης. Ωστόσο παρουσιάζει μεγάλη υπολογιστική πολυπλοκότητα και αυξημένες απαιτήσεις σε μνήμη. Στα πλαίσια της εργασίας αυτής εγίνε μια προσπάθεια ανάλυσης των τεχνικών που βασίζονται στη μέθοδο των συζυγών παραγώγων (Conjugate Gradient), χρησιμοποιούνται σε προβλήματα προσαρμοστικού φιλτραρίσματος και πιο ειδικά στο πρόβλημα της προσαρμοστικής ισοστάθμισης διαύλου. Οι τεχνικές αυτές επεξεργάζονται τα δεδομένα και ανά μπλοκ. Είναι ικανές να παρέχουν ιδιότητες σύγκλισης συγκρίσιμες με αυτές της (RLS) μεθόδου, εισάγοντας υπολογιστική πολυπλοκότητα ενδιάμεσων απαιτήσεων μεταξύ των μεθόδων LMS και RLS χωρίς να παρουσιάζουν προβλήματα αριθμητικής ευστάθειας. / The use of iteration methods for adaptive equalization has received considerable attention during the past several decades. The Least Mean Squares (LMS) method, which has found widespread use owing to its simplicity, has poor convergence properties. The Recursive Least Squares (RLS) method possess superior convergence properties, but it is computationally intensive and has high storage requirements for matrix manipulations. In this MSc thesis the technique of conjugate gradients is applied for the adaptive filtering problem. Conjugate gradient algorithms for adaptive filtering applications suitable for efficient implementation has been developed and has been applied for the design of an adaptive transversal equalizer. Low cost block algorithms using the preconditioned conjugate gradient method are also discussed. The algorithms are capable of providing convergence comparable to RLS schemes at a computational complexity between the LMS and the RLS methods and does not suffer from any known instability problems.