Cette thèse est composée de deux parties indépendantes. Dans la première, nous donnons une démonstration alternative du théorème d'aplatissement par éclatements de Raynaud-Gruson. Celle-ci repose sur la construction et l'étude de certains espaces valuatifs, et nous permet de dégager la notion de $Phi$-anneau, qui fournit un substitut algébrique aux anneaux topologiques adiques : la notion correspondante de $Phi$-schéma est aux schémas ce que les espaces rigides sont aux schémas formels.Dans une seconde partie, nous nous inspirons de travaux de Laumon et de Deligne pour démontrer l'existence de facteurs $varepsilon$ locaux dans un cadre géometrique. Nous démontrons ensuite, en usant de la méthode la phase stationnaire $ell$-adique, une formule du produit pour le déterminant de la cohomologie d'un faisceau $ell$-adique sur une courbe en caractéristique $p neq ell$ positive : cela étend des résultats précédemment connus pour un corps de base fini. Parmi les outils utilisées figure la théorie du corps de classes géométrique, dont nous donnons une démonstration géométrique s'inspirant de l'approche de Deligne pour le cas non ramifié. / This thesis is divided in two parts. We first give an alternative proof of the Raynaud-Gruson's theorem regarding flattening by blow-ups. The argument rests upon the study of certain valuative spaces associated to a refined notion of ring, which we name $Phi$-rings : these are algebraic substitutes to adic topological rings, and the corresponding $Phi$-schemes can be considered as generic fibers of schemes, in the same way that rigid spaces are generic fibers of formal schemes.In the second part, we prove the existence of local $varepsilon$-factors in a geometric setting. These results, which are inspired by works of Laumon and Deligne, lead to a product formula for the determinant of the cohomology of an $ell$-adic sheaf on a curve over a perfect field of positive characteristic $p neq ell$, which was previously known for a finite base field. One of our main tools is geometric class field theory; we provide a detailed proof of its global version by extending Deligne's approach from the tamely ramified case to the general case.

Une ligne de recherche présente dans la littérature depuis les années soixante est celle des \emph{théorèmes de représentation}. Son résultat fondateur est le théorème de Chomsky--Schützenberger qui stipule qu'un langage est algébrique si et seulement si il est l'image par homomorphisme de l'intersection entre un langage régulier et le langage de Dyck. Ce résultat a connu depuis diverses généralisations à différentes familles de langages. Dans cette thèse, nous proposons plusieurs généralisations de ce résultat aux langages d'ordres supérieurs. En particulier, nous introduisons une notion de langages de Dyck d'ordres supérieurs, nous définissons et étudions des classes de transductions que nous qualifions d'$\varepsilon$-sûres et nous montrons qu'un langage appartient à un niveau $k+l$ de la hiérarchie des ordres supérieurs si et seulement si il est l'image d'un langage de Dyck de niveau $k$ par une transductions $\varepsilon$-sûre de niveau $l$. Ces résultats nous permettent aussi d'obtenir d'autres types de caractérisations tels que des caractérisations logiques. / Amongst the classical results of the language theory, one can cite the known characterization of algebraic languages proved by Chomsky and Schützenberger and which states that a language is algebraic if and only if it is the homomorphic image of a regular set intersected with the Dyck language. This result has opened a new line of research and defined a new type of characterizations known as \emph{representation theorems}. In this thesis, we prove various representation theorems for the higher order languages hierarchy. In particular, we introduce a notion of higher order Dyck languages and a hierarchy of classes of transductions that we call $\varepsilon$-stable (or $\varepsilon$-safe) transductions and we prove that a language belongs to some level $k+l$ of the higher order hierarchy if and only if it can be represented as the image of a level-$k$ Dyck language by a level-$l$ $\varepsilon$-stable transduction. These representations also allow us to approach other types of characterizations such as logical characterizations.

acase@tulane.edu

Mixing between overflows and ambient water masses is a crucial problem of deep-water formation in the down-welling branch of the meridional overturning circulation of the ocean. In this dissertation work, performance of second-order turbulence closures in reproducing mixing of overflows is investigated within both hydrostatic and non-hydrostatic models. First, a 2D non-hydrostatic model is developed to simulate the Red Sea overflow in the northern channel. The model results are compared to the Red Sea Outflow Experiment. It is found that the experiments without sub-grid scale models cannot reproduce the basic structure of the overflow. The k-ε model yields unrealistically thick bottom layer (BL) and interfacial layer (IL). A new technique so-called very large eddy simulation (VLES) which allows the use of k-ε model in non-hydrostatic models is also employed. It is found that VLES results the most realistic reproduction of the observations. Furthermore, the non-hydrostatic model is improved by introducing laterally average terms, so the model can simulate the constrictions not only in the z-direction but also in the y-direction. Observational data from the Bosphorus Strait is employed to test the spatially average 2D non-hydrostatic model (SAM) in a realistic application. The simulations from SAM with a simple Smagorinsky type closure appear to be excessively diffusive and noisy. We show that SAM can benefit significantly from VLES turbulence closures. Second, the performance of different second-order turbulence closures is extensively tested in a hydrostatic model. Four different two-equation turbulence closures (k-&epsilon, k-&omega, Mellor-Yamada 2.5 (MY2.5) and a modified version of k- &epsilon) and K-Profile Parameterization (KPP) are selected for the comparison of 3D numerical simulations of the Red Sea overflow. All two-equation turbulence models are able to capture the vertical structure of the Red Sea overflow consisting of the BL and IL. MY2.5 with Galperin stability functions produce the largest salinity deviations from the observations along two sections across the overflow and the modified k-&epsilon exhibits the smallest deviations. The rest of the closures fall in between, showing deviations similar to one another. Four different closures (k- &epsilon, k-&omega, MY2.5KC and KPP) are also employed to simulate the Mediterranean outflow. The numerical results are compared with observational data obtained in the 1988 Gulf of Cadiz Expedition. The simulations with two-equation closures reproduce the observed properties of the overflow quite well, especially the evolution of temperature and salinity profiles. The vertically integrated turbulent salt flux displays that the overflow goes under significant mixing outside the west edge of the Strait of Gibraltar. The volume transport and water properties of the outflow are modified significantly in the first 50 km after the overflow exits the strait. The k-&epsilon and k-&omega cases show the best agreement with the observations. Finally, the interaction between the Red Sea overflow and Gulf of Aden (GOA) eddies has been investigated. It is found that the overflow is mainly transported by the undercurrent at the west side of the gulf. The transport of the overflow is episodic depending strength and location of GOA eddies. The most crucial finding is that the Red Sea overflow leaves the Gulf of Aden in patches rather than one steady current. Multiple GOA eddies induce lateral stirring, thus diapycnal mixing of the Red Sea outflow.

Myoclonus Dystonia (MD) is an autosomal dominant movement disorder characterized by bilateral myoclonic jerks paired with dystonia 1. Mutations have been mapped to the ε-sarcoglycan (SGCE) gene in about 40% of patients 2,92. The purpose of this project was to examine the properties of SGCE in the central nervous system (CNS) and use this knowledge to elucidate the pathology of MD. Although Sgce is a member of the sarcoglycan complex (SGC) in other tissues, little is known about its interactions in the CNS. The vast majority of mutations in SGCE alter the translational reading frame. Proteins arising from these rare mutations are less stable than the wild type (WT) and undergo preferential degradation via the ubiquitin proteasome system 3. As this locus is maternally imprinted, patients with MD are effectively null for sgce expression 73,91. Therefore, Sgce knock out (KO) models should approximate MD conditions both in vivo and in vitro. As there are no current treatments for MD, in sight into the pathology of the disease will aid in eventual treatments and help bring patients some relief by finally understanding their disease. Since a large percentage of MD patients are without the sgce protein, identifying what this protein’s function is and how its absence effects normal processing in the brain should help to identify the underlying cellular pathology which produces the MD phenotype. This research was performed under the hypothesis that, in neuronal cells, sgce interacts with a group of proteins that together play a role in stabilization and localization of ion channels and signaling proteins at the cell membrane. The aims were to: (1) Build a MD mouse model with either a conditional knock-out (cKO) or a conditional gene repair (cGR) mutation; (2) Use neuroblastoma cells to identify the other proteins which interact with sgce in neurons, and; (3) Determine if there is a disruption of the localization of the sgce-complex members due to the loss of sgce. Recombineering was used to complete the constructs for two transgenic mouse models: One model for the KO of exon 4 of sgce and one for the cGR in intron 1. Primary neurosphere lines from two previously generated chimeras were developed, as well as from a WT mouse. These neurosphere cell lines allowed comparisons of RT-PCR results from a heterogeneous neurological cell population to neuroblastoma cell lines. mRNA is present in neuronal cells for many of the DGC associated proteins. It was confirmed that the KD of sgce results in a reduction of nNOS protein and in increased proliferation of NIE cells. By using a nitrite/nitrate assay as well as studies with L-NAME, it was confirmed that this increased proliferation was in fact due to a lack of nNOS function. These proliferation changes did not occur in N2A cells, which do not express high levels of nNOS during proliferation, further confirming nNOS’s role in the proliferation changes. Using qRT-PCR, KD of sgce was shown to result in significant changes in the transcript levels for many DGC associated proteins. This suggests that a DGC-like complex is forming in neuronal cells. Also, as a result of difficulties with the research, it became clear that over-expression of sgce causes cell death. This observation was quantified using cell counts and TUNEL staining, both showing significant results. Additionally, several new constructs were created which will hopefully be of use for future students wanting to study sgce’s functions. New shRNA targeting sgce and sgcb have been made and both constructs result in reducing the expression of sgce. Seven different flag-tagged sgces have been created and some of these have been transferred into a tet-inducible system, which should circumvent the problem of over-expression. Finally GFP-tagged constructs for sgce and sgcb have been made and pooled clones have been developed. These tools will hopefully enable future students to continue to tease apart sgce’s function(s).

Cardiac ischemic preconditioning (IPC) is defined as brief periods of ischemia and reperfusion that protect the heart against longer ischemia and reperfusion. IPC triggers Cl- efflux and protein kinase C epsilon (PKCe) translocation to the particulate fraction. Chloride channel 2 (ClC-2) is volume regulated and is a potential end effector of IPC. The goal of my study was to investigate the involvement of PKCε and ClC-2 protein module in IPC of isolated adult rabbit ventricular myocytes. Co-immunoprecipitation (co-IP) assays on HEK 293 cells, transfected with ClC-2-Flag, confirmed that ClC-2 interacts with PKCe. Subcellular fractionation showed that PKCe/ClC-2 protein module is localized to the sarcolemma of cardiomyocytes. Lastly, ischemia/reperfusion injury was simulated in cardiomyocytes with 45min simulated ischemia (SI)/60min simulated reperfusion (SR) and IPC was induced by pre-treatment with 10min SI/20min SR. Co-IP after each time interval showed that IPC transiently enhanced PKCe/ClC-2 interaction. PKC inhibitor, GF109203X, abrogated the enhanced interaction.

Cardiac ischemic preconditioning (IPC) is defined as brief periods of ischemia and reperfusion that protect the heart against longer ischemia and reperfusion. IPC triggers Cl- efflux and protein kinase C epsilon (PKCe) translocation to the particulate fraction. Chloride channel 2 (ClC-2) is volume regulated and is a potential end effector of IPC. The goal of my study was to investigate the involvement of PKCε and ClC-2 protein module in IPC of isolated adult rabbit ventricular myocytes. Co-immunoprecipitation (co-IP) assays on HEK 293 cells, transfected with ClC-2-Flag, confirmed that ClC-2 interacts with PKCe. Subcellular fractionation showed that PKCe/ClC-2 protein module is localized to the sarcolemma of cardiomyocytes. Lastly, ischemia/reperfusion injury was simulated in cardiomyocytes with 45min simulated ischemia (SI)/60min simulated reperfusion (SR) and IPC was induced by pre-treatment with 10min SI/20min SR. Co-IP after each time interval showed that IPC transiently enhanced PKCe/ClC-2 interaction. PKC inhibitor, GF109203X, abrogated the enhanced interaction.

Polymorphonuclear neutrophils (PMNs) are important effector cells in host defense and the inflammatory response to antigen. The involvement of PMNs in inflammation is mainly mediated by the Fc receptor family, including IgE receptors. Recently, we have shown that human PMNs from allergic asthmatic subjects express the high affinity receptor, FceRI. In this study, we have examined the regulation of FceRI by human PMNs in vitro and in vivo during the allergic pollen season. First we studied the pattern of expression of FceRI in PMNs during the pollen allergic and outside the pollen season. Peripheral blood neutrophils were isolated from adult atopic asthmatics (AA) (n=17), allergic non asthmatics (ANA) (n=15) and healthy donors (n=16) by dextran, ficoll gradient centrifugation and magnetic cell sorting (MACS). Surface, total protein and mRNA expression of FceRI were investigated in the three groups by FACS, immunocytochemistry (ICC) and fluorescent in situ hybridization (FISH) respectively. Secondly, we investigated the effect of Th-2 cytokines which are known to regulate IgE receptor expression. PMNs from atopic asthmatic subjects were stimulated in vitro with Th-2 cytokines (IL-4, IL-9, GM-CSF) and Th-1 cytokine IFN-gamma. Finally we determined whether the expression of FceRIbeta chain correlated with the surface expression of FceRIalpha chain in PMNs. Irrespective of the season, PMNs from atopic asthmatic subjects showed increased expression of FceRIalpha chain in surface, total protein and mRNA compared to atopic non asthmatics and healthy donors (n=20). Interestingly, FceRIalpha chain surface and mRNA expression increased significantly during pollen season compared to non pollen season (P=0.001) in PMNs isolated from AA (n=9) in contrast to healthy donors and ANA (n=8). Furthermore similar pattern of FceRI expression were observed in vitro when PMNs were stimulated with Th2 cytokines. IL-4, IL-9 and GM-CSF showed increased protein and mRNA expression of FceRIalpha chain at 6 and 18hrs (n=6) whereas IFN-gamma down regulated the mRNA expression of FceRIalpha chain at 6hrs. Also, irrespective of season AA (n=11) subjects showed increased expression of FceRI beta chain when compared to ANA (n=10) and healthy donors (n=9). Western blot analysis showed increased FceRI beta protein in atopic asthmatic subjects (n=4). Interestingly irrespective of the groups, there was a positive correlation r = 0.8054 between total protein expression of beta chain with surface expression of alpha chain of FceRI in neutrophils. Our data suggest that the expression of FceRI in neutrophils of atopic asthmatic patients is highly regulated. Our in vitro studies provide evidence that Th-2 cytokines such as IL-9, IL-4 and GM-CSF up-regulate the expression of FceRI. Furthermore we show evidence of increased expression of FceRIbeta chain in neutrophils of atopic asthmatic subjects. Collectively these results suggest that FceRI mediated neutrophil dependent activation may play a key role in allergic diseases. / May 2005

Cette thèse porte sur l'étude de systèmes de type gaz particules, tant d'un point de vue mathématique que physique et numérique. Par ailleurs, quelques aspects de la turbulence en lien avec ces systèmes et le modèle k- sont étudiés.

Polymorphonuclear neutrophils (PMNs) are important effector cells in host defense and the inflammatory response to antigen. The involvement of PMNs in inflammation is mainly mediated by the Fc receptor family, including IgE receptors. Recently, we have shown that human PMNs from allergic asthmatic subjects express the high affinity receptor, FceRI. In this study, we have examined the regulation of FceRI by human PMNs in vitro and in vivo during the allergic pollen season. First we studied the pattern of expression of FceRI in PMNs during the pollen allergic and outside the pollen season. Peripheral blood neutrophils were isolated from adult atopic asthmatics (AA) (n=17), allergic non asthmatics (ANA) (n=15) and healthy donors (n=16) by dextran, ficoll gradient centrifugation and magnetic cell sorting (MACS). Surface, total protein and mRNA expression of FceRI were investigated in the three groups by FACS, immunocytochemistry (ICC) and fluorescent in situ hybridization (FISH) respectively. Secondly, we investigated the effect of Th-2 cytokines which are known to regulate IgE receptor expression. PMNs from atopic asthmatic subjects were stimulated in vitro with Th-2 cytokines (IL-4, IL-9, GM-CSF) and Th-1 cytokine IFN-gamma. Finally we determined whether the expression of FceRIbeta chain correlated with the surface expression of FceRIalpha chain in PMNs. Irrespective of the season, PMNs from atopic asthmatic subjects showed increased expression of FceRIalpha chain in surface, total protein and mRNA compared to atopic non asthmatics and healthy donors (n=20). Interestingly, FceRIalpha chain surface and mRNA expression increased significantly during pollen season compared to non pollen season (P=0.001) in PMNs isolated from AA (n=9) in contrast to healthy donors and ANA (n=8). Furthermore similar pattern of FceRI expression were observed in vitro when PMNs were stimulated with Th2 cytokines. IL-4, IL-9 and GM-CSF showed increased protein and mRNA expression of FceRIalpha chain at 6 and 18hrs (n=6) whereas IFN-gamma down regulated the mRNA expression of FceRIalpha chain at 6hrs. Also, irrespective of season AA (n=11) subjects showed increased expression of FceRI beta chain when compared to ANA (n=10) and healthy donors (n=9). Western blot analysis showed increased FceRI beta protein in atopic asthmatic subjects (n=4). Interestingly irrespective of the groups, there was a positive correlation r = 0.8054 between total protein expression of beta chain with surface expression of alpha chain of FceRI in neutrophils. Our data suggest that the expression of FceRI in neutrophils of atopic asthmatic patients is highly regulated. Our in vitro studies provide evidence that Th-2 cytokines such as IL-9, IL-4 and GM-CSF up-regulate the expression of FceRI. Furthermore we show evidence of increased expression of FceRIbeta chain in neutrophils of atopic asthmatic subjects. Collectively these results suggest that FceRI mediated neutrophil dependent activation may play a key role in allergic diseases.