Human essential hypertension affects over 75 million people in the United States, and can lead to death due to its several serious health complications such as hypertension-related cardiovascular disease. The purpose of this research was to understand how hypertension could cause physiological changes to the microcirculation, specifically the PO2 dependence of oxygen consumption (VO2) in skeletal muscle of normotensive and hypertensive rats. The Spontaneously Hypertensive Rat (SHR) strain was used as the diseased model, and Wistar-Kyoto (WKY) rats were used as controls to conduct this study. The SHR strain develops hypertension between 5-6 weeks after birth with an average systolic blood pressure of 150 mmHg. By arresting blood flow using an objective-mounted inflatable airbag, PO2 measurements were obtained along with an oxygen disappearance curve (ODC), which was used to calculate VO2 over various ranges of physiological PO2 values. PO2 and VO2 curves were analyzed based on Hill’s equation to fit the data and describe the PO2 dependence of VO2. When compared to the healthy Wistar-Kyoto rats, the SHRs exhibited a higher Vmax, or maximum rate of oxygen consumption. The average maximal rate of consumption by the hypertensive animal models could be a consequence of a “mitochondrial uncoupling” or some disconnect in the mitochondrial oxygen consumption and the normal corresponding ATP production. In conclusion, this project demonstrated that in situ muscle tissue from hypertensive and normotensive rats had a PO2 dependence of oxygen consumption over a wide range of physiological PO2 values and the hypertensive rats consumed oxygen at a higher maximal rate.

Ce mémoire présente une exploitation additionnelle du catalogue LSPM comportant deux grandes parties, soit la poursuite du relevé des naines blanches à l'intérieur d'un rayon de 40 pc et l'étude de la contrepartie SDSS de ce catalogue. La première consiste à utiliser les critères basés sur des diagrammes de mouvements propres réduits établis dans des travaux antérieurs afin de dresser une liste de candidates naines blanches. Quelques modifications quant à l'ordre de priorité ainsi que le critère basé sur les magnitudes photographiques seront apportées. Une approche de moindres carrés non-linéaire appliquée aux magnitudes observées dans les systèmes photométriques disponibles permet ensuite de déterminer la distance des objets sélectionnés par comparaison avec les plus récents modèles de naines blanches disponibles. Un second critère est appliqué en se basant sur la qualité de l'ajustement résultant de la procédure. Ceci mènera à l'identification de 31 nouvelles naines blanches, dont 11 situées vraisemblablement à moins de 40 pc. Une nouvelle liste de candidates a aussi pu être établie pour la poursuite du relevé. La seconde partie consiste à utiliser tous les objets du 7e relevé du SDSS identifiés dans le catalogue LSPM pour l'étude statistique des naines blanches, essentiellement via la détermination de leur fonction de luminosité. Pour ce faire, un critère basé sur les diagrammes de mouvements propres réduits sera encore une fois utilisé. La distance des objets sélectionnés sera déterminée selon la même procédure, mais cette fois en ne se limitant pas à 40 pc. La méthode de pondération selon le volume observable (1/v_max) est utilisée afin de compenser pour le biais introduit par la sensibilité du catalogue à la magnitude. Cependant, d'autres facteurs viennent influencer la détermination de la fonction de luminosité et une analyse de ceux-ci est finalement présentée. / We present an additional exploitation of the Lépine \& Shara Proper Motion catalog (LSPM) divided in two main parts: a follow up and some improvements on the census or northern white dwarfs within 40 pc of the Sun and a study of the SDSS white dwarfs component in the LSPM survey. The former consists in the use of criteria previously established in order to create a list of white dwarf candidates with an associated priority. The priority order has been enriched and one of the criteria has been slightly modified. We then use a non-linear least square method to the observed magnitudes in each available photometric system simultaneously in order to determine the atmospheric parameters and, in particular, the distance of each white dwarf candidate. This approach allows a second criteria to be applied on our sample based on the goodness of the fit. This will lead us to the identification of 31 new white dwarfs, from which 11 are likely to remain within 40 pc of the Sun. A new list of 340 candidates has also been established for eventual observations. The latter consists in the use of all the objects from the 7th data release of the SDSS that have a counterpart in the LSPM catalog to elaborate a statistical study of white dwarfs, in this case by calculating the white dwarf luminosity function. To do so, one of the same criterion mentioned above will be used to make a first selection of presumed white dwarfs. Afterwards, the distance will be determined by the same least-square method, but without any restriction on the distance. To balance the effect due to the sensibility in magnitude of the survey, we used a ponderation method based on the maximum observable volume (1/v_max). However, other factors come to affect our results and the last part of this work concentrates on their identification.

The elimination of drugs from the body is in many cases performed by the liver. Much could be gained if an accurate prediction of this process could be made early in the development of new drugs. However, for the elimination to occur, the drug molecule needs first to get inside the liver cell. Disposition is the expression used to encapsulate both elimination and distribution. This thesis presents novel approaches and models based on simple in vitro systems for the investigation of processes involved in the hepatic drug disposition. An approach to the estimation of enzyme kinetics based on substrate depletion data from cell fractions was thoroughly evaluated through experiments and simulations. The results that it provided were confirmed to be accurate and robust. In addition, a new experimental setup suitable for a screening environment, i.e., for a reduced number of samples, was generated through optimal experimental design. The optimization suggested that sampling at late time points over a wide range of concentration was the most advantageous. A model, based on data from primary hepatocytes in suspension, for the investigation of cellular disposition of metabolized drugs was developed. Information on the relative importance of metabolism and membrane protein related distribution was obtained by analysis of changes in the kinetics by specific inhibition of the various processes. The model was evaluated by comparing the results to those obtained from an in vivo study analyzed with an especially constructed mechanistic PBPK model. These investigations showed that the suggested model produced good predictions of the relative importance of metabolism and carrier mediated membrane transport for hepatic disposition. In conclusion, new approaches for the investigation of processes involved in hepatic disposition were developed. These methods were shown to be robust and increased the output of information from already commonly implemented in vitro systems.

La recherche de variations d’activités enzymatiques associées à des phénotypes chez la tomate (Solanum Lycopersicum) a permis d’apporter de nouveaux éléments pour l’étude desrelations existantes entre le métabolisme central et la qualité du fruit. Ce projet qui engageaitune nouvelle thématique au sein de l’unité Biologie et Pathologie de Fruit (UMR 1332, INRABORDEAUX) a permis dans un premier temps de développer une plateforme d’enzymologieà haut débit. Cette structure permet d’une part de réaliser plus de 10 000 déterminationsd’activités enzymatiques par jour avec une très grande reproductibilité et d’autre part dedéterminer les constantes de Michaelis (Km) apparentes pour jusqu’à une dizaine d’enzymespar jour.Dans un deuxième temps ce projet s’est attaché à étudier les relations existantes entre lesenzymes de la voie de la glycolyse chez le cultivar de tomate MicroTom. Nous y avonsrelevé l’existence de corrélations fortes entre les activités de ces enzymes. Nous avonségalement mis à jour l’existence de corrélations pour les enzymes mesurées à partir de deuxétages foliaires distincts ce qui suggère que les réseaux enzymatiques sont conservés ausein d’une plante.Dans un troisième temps, le criblage d’une collection de mutants de tomate MicroTom a étéentrepris. Ce criblage de plus de 150 familles (soit environ 1800 plantes) sur la base desactivités enzymatique de onze enzymes du métabolisme central à deux concentrations desubstrats différentes (saturante et non saturante) a abouti à l’identification de deux familles.Ces deux familles porteraient chacune une mutation affectant les caractéristiques cinétiquesde la Triose-Phosphate Isomérase. Ces mutations étaient toujours en cours d’étude à la finde ce projet. Ces résultats ouvrent de nouvelles perspectives pour la compréhension desrelations entre les enzymes du métabolisme central. Ils permettent aussi d’apporter desméthodes d’identification rapide de mutants enzymatiques au sein d’une large population. / Research on enzymatic variations associate with phenotypes in tomato (SolanumLycopersicun) provided new and original input regarding links between central metabolismand fruit quality. This project took part in a new topic of the Fruit Biology and Pathology Unit(UMR 1332, INRA BORDEAUX). First, during this project, a new high-throughputenzymology platform was created. This unique lab offers possibility to determine both morethan 10 000 enzyme activities per day with a very good reliability and apparent Michealisconstant (Km) for up to ten enzymes per day.Second, this project investigated existent relationship between glycolytic enzymes inMicroTom tomato cultivar. We highlighted strong correlations between enzymes in leaves.We also uncovered correlations between enzymes activities measured in two distinct foliarlevels. These elements suggest inheritability of the enzymes network within the plant.Third, the screen of an Ethylmethyl Sulfonate (EMS) MicroTom mutant’s collection wasinitiated. 150 families (around 1800 plants) were screened for eleven enzymes with twodifferent substrate concentrations. At the end of the process, two families were identified;both could have mutation(s) that affect(s) the kinetic characteristic of Triose-Phosphateisomerase. These mutations were style investigated at the end of this project. These originalresults provide new perspectives for knowledge of relationship between central metabolism’senzymes. Finally, This project proposes new and rapid enzymatic mutant identification withina large population as an EMS mutant’s collection.