Die Arbeit umfasst die Synthese von lipophilen Inositolen und Glycerollipiden, welche auf ihre Raftophilie getestet wurden. Des weiteren wurden eine Reihe neuer Ceramide synthetisiert und diese in Bioassays auf ihre Wirksamkeit gegenüber diversen Krankheiten wie Influenza getestet.

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Plasmodium falciparum est responsable de la forme la plus grave de paludisme avec plus de 600 000 décès par an. L'absence de vaccin efficace, combinée à l'émergence de résistances aux traitements récurrents, exige le développement de nouvelles molécules. Afin de limiter ces résistances, il est nécessaire de cibler de nouvelles voies métaboliques indispensables à la survie du parasite. Ce travail de thèse repose sur l'étude de deux voies métaboliques essentielles au parasite que sont la voie de recyclage des bases puriques et la voie de biosynthèse des ancres glycosylphosphatidylinositol (GPI).En ce qui concerne la voie de recyclage des bases puriques, la détermination des structures cristallines de l' « IMP specific 5‘-nucleotidase » (PfISN1) associée aux études biochimiques et biophysiques (SAXS, EM, MALS…), a permis de préciser le mécanisme d'action fournissant ainsi une base solide pour la mise au point d'inhibiteurs. Une banque de plus 3000 composés a été criblée par Fluorimétrie à Balayage Différentiel et les effets des molécules sélectionnées seront évalués sur l'enzyme et sur la croissance du parasite en culture.Quatre cibles thérapeutiques potentielles appartenant à la voie de biosynthèse des ancres GPI ont été sélectionnées. L'utilisation de plusieurs systèmes d'expression disponibles au laboratoire (bactérie, levure, acellulaire en germe de blé) ou via des plateformes européennes pour l‘expression en cellules de mammifères HEK293T (Oxford), de cellules BHK21 transfectées avec le virus de la vaccine modifié, T7-MVA, (Strasbourg) ou la plateforme ESPRIT (Grenoble) ont permis de passer outre les difficultés rencontrées pour exprimer les protéines d'intérêt. L'une des quatre cibles, la mannose-1-phosphate guanylyltransférase (PfMPG), a pu être exprimée de manière suffisante quantitativement et qualitativement pour une caractérisation biochimique et structurale. Une analyse par SAXS et cristallographie aux rayons X a été réalisée / Plasmodium falciparum is responsible for the most severe form of malaria with more than 600,000 deaths per year. The lack of an effective vaccine, combined with the emergence of drug resistant parasites, necessitates the development of new drugs. In order to limit these resistances, it is necessary to target new metabolic pathways essential for parasite survival. This thesis work is based on the study of two metabolic pathways essential to the parasite, the purine salvage pathways and the glycosylphosphatidylinositol (GPI) anchor biosynthesis pathway.Concerning the purine salvage pathway, the determination of the crystal structures of the IMP -specific 5'-nucleotidase (PfISN1) associated with biochemical and biophysical studies (SAXS, EM, MALS, etc.) have allowed to propose a reaction mechanism, thereby providing a solid basis for the conception and development of inhibitors. A library of more than 3000 compounds was screened by Differential Scanning Fluorimetry and the selected molecules will be evaluated for their inhibitory effect on the enzyme and on the growth of parasites in culture.Four potential therapeutic targets belonging to the GPI anchor biosynthesis pathway were selected. The use of several in-house available expression systems (bacteria, yeast, and acellular wheat germ) as well as European platforms for the expression in HEK293T mammalian cells (Oxford), in BHK21 cells transfected with the modified vaccinia virus, T7-MVA, (Strasbourg) or the ESPRIT platform (Grenoble) has allowed us to overcome the difficulties encountered on obtaining the selected protein targets. One of the four targets has been expressed in sufficient amount and quality for biochemical- and structural characterization, namely the mannose-1-phosphate guanylyltransferase (PfMPG). SAXS and X-ray crystallography analyses have been carried out

As processors' clock-speed flattened out in the early 2000s, multi-core processors became more prevalent and so did parallel programming. However this programming paradigm introduces additional complexities, and to combat this, the SkePU framework was created. SkePU does this by offering a single-threaded interface which executes the user's code in parallel in accordance to a chosen computational pattern. Furthermore it allows the user themselves to decide which parallel backend should perform the execution, be it OpenMP, CUDA or OpenCL. This modular approach of SkePU thus allows for different hardware to be used without changing the code, and it currently supports CPUs, GPUs and clusters. This thesis presents a new so-called SkePU-backend made for clusters, using the communication library GPI. It demonstrates that the new backend is able to scale better and handle workload imbalances better than the existing SkePU-cluster-backend. This is achieved despite it performing worse at low node amounts, indicating that it requires less scaling overhead. Its weaknesses are also analyzed, partially from a design point of view, and clear solutions are presented, combined with a discussion as to why they arose in the first place.

The use of Gross Domestic Product (GDP) as the primary measure of economic progress has arguably led to unintended consequences of environmental degradation and socially skewed outcomes. The Genuine Progress Indicator (GPI) was designed to reveal the trade offs associated with conventional economic growth and to assess the broader impact of economic benefits and costs on sustainable human welfare. Although originally designed for use at the national scale, an interest has developed in the United States in a state-level uptake of the GPI to inform and guide policy. However, questions exist about the quality and legitimacy of the GPI as a composite indicator. These questions include concerns about the underlying assumptions, the monetary weights and variables used, statistical rigor, magnitude of data collection required, and lack of a transparent governance mechanism for the metric. This study aims to address these issues and explore the GPI through a design-thinking lens as both a design artifact and intervention. The leading paper in this dissertation offers the first GPI accounting for all 50 U.S. states. State GPI results are introduced and compared to Gross State Product (GSP). Then an analysis of the components to GPI reveals which drive the differences in outcomes, including examining the sustainability aspects of the state-level results. The second paper investigates the quality of the GPI as a composite indicator by testing its sensitivity to numerical assumptions and relative magnitudes of components, with particular attention to the possible unintended policy consequences of the design. The third paper seeks to answer the question of both efficiency (data parsimony) and effectiveness (comparatively to other indicators) by analysis of correlations between GPI components and with other state-level indicators such as the Gallup Well-Being Indicator, Ecological Footprint, and UN Human Development Index. To garner insight about possible GPI improvements, goals, and governance gaps in the informal U.S GPI network, the final paper dives into processes, outputs, and outcomes from the community of practice as revealed through a facilitated U.S. GPI workshop.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-10-03T11:28:45Z No. of bitstreams: 2 Dissertação - Jordana Fernandes de Castro - 2013.pdf: 4456871 bytes, checksum: ad2725a858a3d067967c1a37a5307aaf (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-10-06T10:37:13Z (GMT) No. of bitstreams: 2 Dissertação - Jordana Fernandes de Castro - 2013.pdf: 4456871 bytes, checksum: ad2725a858a3d067967c1a37a5307aaf (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-06T10:37:13Z (GMT). No. of bitstreams: 2 Dissertação - Jordana Fernandes de Castro - 2013.pdf: 4456871 bytes, checksum: ad2725a858a3d067967c1a37a5307aaf (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-08-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Anthropological studies pertaining to dams in Brazil, from the 1970s, their numbers have considerably increased concomitantly increasing the works of the Brazilian Electricity Sector, which has a prominent place in development strategies designed by the National State. The creation, from the 1940s, the regional development plans, where the regions are to be seen as areas of action and state control for national economic growth through an internal colonialism elects as instruments of this policy the Major Projects Investment (GPIs), projects that is based on the exploitation of natural and human resources in economic logic. The exploitation of these resources and the dismantling of the deployment region of GPI generate perverse effects on the populations affected, presenting itself as a consolidated research strand. In this sense, the research presented here was to object eight families relocated for an agricultural community, as a result of dam construction Ribeirão João Leite, Goiânia, Goiás, between the years 2002 and 2009. The objective of the study is to understand the process of moving to an unknown location and the process of social transformation resulting from this relocation. The data presented are the result of the method of participating observation and documentary research in the archives of the Sanitation Company of Goiás S / A - SANEAGO, responsible for the work and analyzed with the help of theoretical texts on the subject of large dams. Keywords: Dam Ribeirão João Leite. Displacement Compulsory. Grande Project Investment (GPI). / Os estudos antropológicos referentes às barragens no Brasil, a partir da década de 1970, têm aumentado consideravelmente seus números concomitantemente às crescentes obras do Setor Elétrico Brasileiro, que ocupam lugar de destaque nas estratégias de desenvolvimento pensadas pelo Estado Nacional. A criação, a partir da década de 1940, de planos de desenvolvimentos regionais, onde as regiões passam a serem vistas como territórios de ação e controle estatal para o crescimento econômico nacional, através de um colonialismo interno, elege como instrumentos dessa política os Grandes Projetos de Investimentos (GPIs), empreendimentos que tem como base a exploração de recursos naturais e humanos sob lógica econômica. A exploração destes recursos e a desarticulação da estrutura da região de implantação do GPI geram efeitos perversos nas populações atingidas, apresentando-se como uma vertente consolidada de pesquisas. Nesse sentido, a pesquisa que aqui se apresenta teve como objeto oito famílias relocadas para uma agrovila, em decorrência da construção da barragem do Ribeirão João Leite, em Goiânia, Goiás, entre os anos de 2002 e 2009. O objetivo do estudo é compreender o processo de mudança para um local desconhecido e o processo de transformação social decorrente desta relocação. Os dados apresentados são resultado da observação participante como método e pesquisa documental nos arquivos da Companhia de Saneamento de Goiás S/A – SANEAGO, responsável pela obra e analisados com apoio de textos teóricos sobre a temática das grandes barragens.

Endocytosis effectuates a critical interface between the eukaryotic cell and its apposing environment. It is, subsequently, paramount for many physiologically important processes and encompasses a diverse array of mechanisms and pathways. The classical endocytic routes mediated by clathrin and caveolin are the best understood and the molecular roles of their major regulators, such as dynamin, adaptor proteins and various lipid species, are the most comprehensively described. Recent identification of an assortment of constitutive, noncaveolar, clathrin-independent endocytic (CIE) pathways has expanded the endocytic system. Unlike the classical endocytic pathways, little is known about the guiding parameters of CIE routes. Consequently, it is not possible to understand the important cellular roles these pathways may be fulfilling. This study has begun to characterise the very basic parameters governing the morphologically striking Clathrin-Independent Carrier (CLIC) pathway. Development of a diverse molecular toolkit has now allowed the quantitation of endocytic capacity provided by CLICs, the visualisation of subtle sorting components of the CLIC pathway, the isolation of novel CLIC cargo and regulators, and has linked this mechanism to the critical cellular processes of cellular migration and membrane repair. Calculation of the individual capacity of endocytic routes provides important information about the contribution of each pathway to total plasma membrane (PM) uptake and turnover. Quantitation of the volume, surface area and number of structures forming per minute in this study shows that CLICs provide the vast majority of constitutive endocytosis, up to four times the capacity of the clathrin mediated endocytic (CME) pathway. As the equivalent of the entire PM area could pass through the CLIC pathway within 12 minutes it is evident that CLICs are fundamental housekeepers of bulk membrane internalisation. Thus, they are likely to be central regulators of PM homeostasis and turnover. High-resolution tomography, in conjunction with analysis of CLIC cargo trafficking, identifies these carriers as complex, pleiomorphic structures that sort the bulk of membrane to early endosomes and recycle cargo back to the cell surface. Such vast internalisation combined with an ability to rapidly recycle components quickly attributes the CLIC pathway as a complex sorting station. Isolation of novel cargo and regulators has identified a striking array of proteins now associated with the CLIC pathway for the first time. A significant proportion of identified targets localise to lipid-rafts and recycle from the PM, facets consistent with association to the CLIC pathway. Numerous targets have also been directly implicated in clathrin-independent endocytosis by independent groups. Verification of selected cargo, such as CD44, Thy-1 and myoferlin, showing specific internalisation through the CLIC pathway, has provided insight into the sorting ability of the CLIC pathway and links to adhesion turnover and membrane recycling. Consistent with a role in cellular adhesion turnover, it was found that CLICs become polarised within migrating cells. This has shown the first instance of spatial separation between three major endocytic routes, CLICs, caveolae and CME and highlights the important and coordinated roles of multiple endocytic pathways during physiologically significant processes. The specific internalisation of paxillin, Thy-1 and CD44 through CLICs at the leading edge of migrating cells suggests that CLICs rapidly turnover adhesion components for dynamic extracellular sensation during directional cell migration. Indeed, specific ablation of the CLIC pathway significantly impedes cellular migration, implying coordination with CME at the leading edge. This study has defined numerous parameters of the CLIC pathway, developing the current understanding of this poorly defined route and places the CLIC pathway as a unique player during critical cellular processes.

La paroi du champignon filamenteux A. fumigalus conditionne la croissance et est responsable du maintien de l'intégrité cellulaire lors de l'infection. Les protéines de la paroi de ce champignon filamenteux n'avaient pas été étudiées jusqu'à présent alors qu'elles semblent jouer un rôle essentiel dans la structuration de la paroi de la levure modèle S. cereviciae. Une approche essentiellement biochimique a permis de caractériser les protéines associées à la paroi de A..fumigatus. La majorité des protéines pariétales chez A. fumigatus sont des protéines solubles. Une seule protéine est relarguée à partir de la paroi par un traitement f3- (1 .3) glucanase : c'est une phosphatase acide qui possède une ancre GPl et dont l'expression est réprimée en présence de phosphate inorganique. Par ailleurs, une étude des protéines GPl chez A. fumigatus par génomique comparative a montré que les protéines GPI décrites comme associées de façon covalente à la paroi chez la levure n'ont pas d'homologue chez A. fumigatus. Ainsi. l'organisation des protéines au sein de la paroi de A..fumigatus est différente de celle de la levure : les protéines pariétales ne semblent pas avoir un rôle essentiel dans l'élaboration de la paroi. Ensuite, une nouvelle famille de glycoprotéines portant un N-glycane lié à un galactofuranose en position terminale a été décrite. Cette famille comprend une phospholipase C, une phosphatase alcaline et une phytase. Enfin, une analyse morphologique de deux mutants chitine synthase et gluconosyltransférase a permis d'associer la réduction de la croissance à un hyper-branchement du mycélium et à une diminution de la taille de la cellule apicale sans que l'organisation globale des polysaccharides pariétaux et le taux de croissance spécifique soient affectés.

Epithelial cells represent the ability to polarize with an apical and basolateral domains which differ markedly in proteins, lipid composition and therefore in function. This asymmetry reflects the ability of epithelial cells to sort newly synthesized proteins and lipid to either cell surface. While the signals responsible for basolateral targeting of the proteins have been clearly understood, the situation regarding the apical sorting of proteins is more obscure. We have previously shown that differently from basolateral GPI-APs oligomerization in the Golgi apparatus is necessary for apical sorting of Glycosylphosphatidylinositol- anchored proteins (GPI-APs). Interestingly this mechanism is conserved in two different kinds of epithelial cells, MDCK and FRT cells, which exhibits a difference in the sorting of GPI-APs. However the precise mechanism leading to this event is not understood. Our previous data demonstrated that simple addition of cholesterol to MDCK cells is necessary and sufficient to induce the oligomerization and apical sorting of a basolateral GPI-AP. Whereas, in this present study in FRT cells we showed that in contrast with MDCK cells cholesterol is not an active player in the regulation of GPI- APs apical sorting. In addition, we also showed that apical and basolateral GPI-APs are not segregated in the Golgi on the bases of the cholesterol content of the surrounding membrane environment. Furthermore, we demonstrated that N- glycosylation of the protein ectodomain is critical for oligomerization and apical sorting of GPI-APs. Our data indicates that at least two mechanisms depending either on cholesterol or on N-glycosylation exist to determine oligomerization in the Golgi and sorting to the apical membrane of GPI-APs.

<p>Abstract</p><p>In order to address sustainable policies, communities need indicators that can tell them something about the larger social and ecological system, and the relationships between the two. This study attempts to apply an existing indicator of welfare and sustainability, the Genuine Progress Index (GPI), to the regional level in Sweden. The county of Östergötland is used as a case study and the thesis is written in collaboration with Regionförbundet Östsam. The aim of the study is to develop an application of GPI methodology to circumstances of data availability at the regional Swedish level. Developed methodology used to calculate a GPI of Östergötland in 2000-2006. GPI results are put in a context of the Regional Development Program of Östergötland in order to address policy implications regarding the very meaning of growth, development, welfare and sustainability. The results indicate that GPI has increased during the time period. However, a comparison with the Gross Regional Product (GRP) shows that GRP has increased faster than GPI. It is concluded that this could be a possible trend of decreased economic efficiency, where more economic output is not sustainable in the sense that an equivalent amount of welfare is not produced. This effect is mainly an effect of increasing income inequality. It is also suggested that the Regional Development Program should distinguish growth and development as different concepts. In doing so, more sustainable policies can be adopted in the future.</p>