Univ., Diplomarbeit, 2002--Frankfurt (Main) / Zsfassung in dt. und engl. Sprache

No description available.

In order to better manage progress toward improved human welfare, governments and organizations around the world have begun to report on more comprehensive indicators of environmental, social, and economic conditions. The Genuine Progress Indicator (GPI) has proven useful as a measure of economic welfare by incorporating changes in environmental conditions, resource stocks, social capital, income distribution, and other non-marketed economic activity. Studies at the local scale have also found the GPI to be an effective tool for informing debate and stimulating questions about the nature of the economic development process. In this study, the GPI methodology is applied to Baltimore City, Baltimore County, and Maryland in order to explore how sustainable economic welfare in the Baltimore region has changed from 1950-2005. A comparison among per capita GPI trends in four US cities shows Baltimore to have the highest average annual growth rate over the study period. Comparisons are made between per capita GPI and Gross Domestic Product (GDP), the most widely recognized measure of national economic performance. Analysis of the trends at all three scales show that GDP growth does not correlate well with changes in welfare as measure by GPI. This implies that Baltimore City, Baltimore County, and Maryland could be in a period of uneconomic growth, when the social and environmental costs of further economic growth outweigh the benefits of such growth. However, the underlying methods used in sub-national applications of the GPI inevitably lead toward certain results, giving rise to an indicator framework that favors particular policy and development outcomes. This situation is defined as indicator bias. Since indicator bias can inadvertently lead society toward undesirable conditions, key assumptions that contribute to indicator bias in the GPI are tested for how they influence the final GPI results. The costs of crime, long-term environmental damage, and depletion of non-renewable natural resources categories are explored in more depth. GPI is found to be an imperfect measure of true progress, but it is believed to be an improvement over GDP for guiding modern society towards a more sustainable and desirable future. More work is needed to incorporate uncertainty, fine-tune the underlying GPI methodology, and build broad consensus about how to measure economic performance and social progress. By providing information about social, ecological, and economic conditions of the region, though, the Baltimore GPI does inform citizens and decision-makers about a wide range of impacts resulting from the modern ‘GDP growth’ paradigm

L'une des infections parasitaires les plus courantes chez les animaux à travers le monde est la babésiose ou piroplasmose. Causée par le développement intraérythrocytaire d'un parasite du genre Babesia, elle présente de nombreux signes cliniques semblables à ceux du paludisme. Ce parasite, du phylum des Apicomplexes, est transmis via le vecteur tique et effectue son cycle de reproduction dans les cellules rouges du sang de l'hôte vertébré. En Europe B. divergens et B. canis sont les espèces majoritairement responsables respectivement de la babésiose bovine et la babésiose canine. Dans une stratégie de recherche vaccinale, l'étude de protéines parasitaires en contact avec la circulation sanguine est primordiale pour comprendre les interactions hôte-parasite et identifier des candidats vaccins à haut potentiel. Les protéines à ancrage GPI (glycosylphosphatidylinositol) font partie de ces protéines. La première protéine à ancrage GPI décrite chez B. divergens est Bd37.1. Elle induit une protection totale contre une infection à B. divergens à la condition qu'une séquence hydrophobe soit ajoutée en C-terminale. La résolution de la structure RMN de cette protéine a permis de mettre en évidence un probable mécanisme de changement conformationnel en fonction du pH. La structure composée de 3 sous domaines montre que celle-ci n'est maintenue que par des ponts salins qui peuvent se rompre en milieu acide. Or l'environnement membranaire dans lequel évolue Bd37.1 ancrée à la surface du parasite et/ou à l'approche du globule rouge lors de l'invasion est acide. Cette dynamique conformationnelle de la protéine Δ-Bd37, liée à l'environnement membranaire, pourrait être à l'origine du mécanisme qui confère une immunité en fonction de la présence ou non de la séquence hydrophobe en C-terminale de Bd37.1. Nous avons cherché à estimer les implications d'une telle dynamique dans les interactions hôtes-parasites à travers l'étude structurale de 2 protéines parasitaires (Bd37.1 et Bc28.1). Dans le premier cas nous étudions la dynamique conformationnelle de la protéine d'adhésion Bd37.1. Nous avons exploré les différentes conformations que pourrait adopter la protéine Bd37.1 par une approche de biophysique et nous avons stabilisé ces différentes conformations en solution par le biais de mutations pour les étudier. Parmi ces mutants, le mutant EDK-Δ-Bd37 dont les ponts salins ont été rompus montre des caractéristiques différentes de Δ-Bd37. Les données enregistrées sur ce mutant nous ont amené à résoudre sa structure et à tester son pouvoir vaccinant. Dans une seconde partie, nous caractérisons biochimiquement et fonctionnellement une autre protéine Bc28.1, l'orthologue de Bd37.1. chez B. canis, accompagnée de la résolution de sa structure. Nous montrons que Bc28.1 est une protéine d'adhésion localisée à la surface du parasite et nous comparons les structures de Bd37.1 et Bc28.1. Ces deux structures sont finalement très différentes tandis que localisation et fonction sont similaires. / One of the most common parasitic infections in animals worldwide is babesiosis or piroplasmosis. Caused by the intraerythrocytic development of Babesia parasite, it has many clinical signs similar to those of malaria. This parasite of the phylum Apicomplexa, is transmitted via the tick vector and performs its reproductive cycle in red blood cells of the vertebrate host. B. In Europe divergens and B. canis species are mainly responsible respectively for bovine babesiosis and canine babesiosis. A strategy of vaccine research, the study of parasite proteins in contact with the bloodstream is essential for understanding host-parasite interactions and identify vaccine candidates with high potential. Anchored protein GPI (glycosylphosphatidylinositol) are part of these proteins. The first protein GPI anchors described in B. divergens is Bd37.1. It induces complete protection against infection with B. divergens provided a hydrophobic sequence is added at the C-terminus. Resolution NMR structure of this protein has highlighted a probable mechanism of conformational change as a function of pH. The structure consists of three sub areas shows that it is only maintained by salt bridges which can break in acidic medium. However, the environment within which Bd37.1 membrane anchored to the surface of the parasite and / or approach the red blood cell during the invasion is acidic. This conformational dynamics of the protein-Δ Bd37 linked to the membrane environment, could be at the origin of the mechanism that confers immunity depending on the presence or absence of the hydrophobic sequence at the C-terminus of Bd37.1. We sought to assess the implications of such dynamics in host-parasite interactions through structural study of two parasite proteins (Bd37.1 and Bc28.1). In the first case we study the conformational dynamics of the adhesion protein Bd37.1. We explored the different conformations that may be adopted by a protein Bd37.1 biophysical approach and we have stabilized in different conformations in solution through mutations to study. Among these mutants, the mutant Δ-Bd37-EDK including salt bridges were broken shows different characteristics Δ-Bd37. The data on this mutant led us to solve the structure and to test its power vaccinating. In a second part, we characterize biochemically and functionally Bc28.1 another protein, the ortholog Bd37.1. in B. canis, accompanied with the resolution of its structure. We show that Bc28.1 is an adhesion protein localized to the parasite surface and compare the structures and Bd37.1 Bc28.1. These two structures are ultimately very different while location and function are similar.

A incidência de infecções por fungos oportunistas na população de pacientes imunocomprometidos tem aumentado nos últimos anos, e estas são, principalmente, causadas por Candida albicans. Este patógeno oportunista pode crescer em diferentes formas, variando de levedura, pseudohifa e hifa e essa transição morfológica está associada com a virulência. Na transição de levedura para hifa, genes hifa-específicos são expressos e muitos deles codificam proteínas que possuem uma molécula de GPI (glicosilfosfatilinositol), mas a maioria (66%) das proteínas preditas ancoradas por GPI (PGAs), ainda possuem função desconhecida. Portanto, o objetivo deste trabalho, foi iniciar a caracterização funcional dos genes PGA13 e PGA58 através de análises in silico, da expressão diferencial dos RNAm sob condições ambientais diversas, e nos mutantes nulos TUP1 e EFG1, e a obtenção e caracterização fenotípica dos mutantes de PGA13: homozigoto nulo e de superexpressão. Os estudos realizados in silico indicam que PGA13 e PGA58 são reguladas pelos fatores transcricionais Efg1, Tec1 e Nrg1 e que possuem sítios potenciais de glicosilação. A análise transcricional desses genes mostra que ambos são regulados por Tup1 e que respondem aos estímulos NaCl (Cloreto de sódio), H2O2 (Peróxido de hidrogênio), etanol, cafeína, HCl (Ácido Clorídrico) e às condições hipo e hiperosmótica. Os mutantes nulos e de superexpressão de PGA13 e PGA58 foram obtidos e as linhagens que perderam o gene PGA13 tiveram um padrão de crescimento da colônia diferente da linhagem CAI-4 e foram mais sensíveis aos compostos SDS, Higromicina B e pH ácido, enquanto o mutante que superexpressava foi mais resistente. Estes resultados sugerem que Pga13 deve ter papel importante na parede celular, visto que a falta dela ocasionou maior sensibilidade a compostos que agridem a parede celular. / The incidence of opportunistic fungal infections in immunocompromised patients has increased in the last years. Candida albicans is the most commonly isolate in immunocompromised patients. C. albicans may grow in distinct morphologies: yeast, pseudohyphal and true hyphal. The switch between yeast and filamentous forms is strongly associated with virulence. During the transition, hyphal-specific genes are expressed and many of these genes encode glycophosphatidylinositol (GPI)- anchored proteins. The majority (66%) of predited GPI proteins has unknown functions. The purpose of this work was to functionally characterize the novel PGA13 and PGA58 genes. To accomplish this task, we have made in silico promoter analysis, mRNA differential expression analysis under some environmental conditions. We have also verified whether those genes are regulated by Tup1 and Efg1 transcriptional regulators. We have knocked out PGA13 and have done phenotypic analysis of the resulting PGA13 null mutant strain and a Pga13 overexpressing strain. The in silico promoter analysis indicates that PGA13 and PGA58 may be regulated by the transcriptional factors Efg1, Tec1 and Nrg1. PGA13 and PGA58 aminoacid sequence analysis revealed that they have potential glycosylation sites. The transcriptional analysis has shown that both genes are regulated by the morphogenesis negative regulator Tup1. PGA13 and PGA58 genes have differential expression in response to salt, hydrogen peroxide, ethanol, caffein, and acid stresses, and also to hypo and hyperosmotic shocks. The phenotypic analysis shows that the pga13/pga13 mutant was more sensitive to SDS, Hygromycin B and acid pH in a plate dilution sensitivity test. Interestingly, the overexpressing mutant strain was more resistant to the same compounds. Taken together these results suggest that PGA13 may have an important role in the cell wall since its absence leads to higher sensitivity to compounds that affect this organelle.

Les cellules épithéliales sont capable de se polariser avec un domaine apical et un basolatéral qui diffèrent nettement en leurs protéines, leurs composition en lipides et donc en fonction. Cette asymétrie reflète la capacité des cellules épithéliales à affecter les protéines nouvellement synthétisées et les lipides à chacune des surfaces de la cellule. Alors que les signaux responsables pour la sélection basolatérale des protéines ont été clairement identifiés, la situation en ce qui concerne la sélection apicale des protéines demeure mal comprise. Nous avons précédemment montré que contrairement aux GPI-APs basolatérales l’oligoisomérisation des protéines ancrées glycosylphosphatidylinositol (GPI-AP) dans l'appareil de Golgi est requise pour le tri apical. Il est intéressent de noter que ce mécanisme est conservé dans deux types de cellules épithéliales, les cellules MDCK et les cellules FRT, qui présentent des différences de tri des GPI-APs. Cependant, le mécanisme précis menant à cet événement n'est pas compris. Nos données précédentes ont démontré que le simple ajout de cholestérol aux cellules MDCK est nécessaire et suffisant pour induire l'oligomérisation et le tri apical d'une GPI-AP basolatérale. Alors que dans l’étude présente sur les cellules FRT, nous avons montré que contrairement aux cellules MDCK, le cholestérol ne joue pas un rôle majeur dans la régulation du tri apicale des GPI-APs. De plus, nous avons également montré que les GPI-APs apicales et basolatérales ne sont pas séparées dans le Golgi en fonction de leur teneur en cholestérol entourant l'environnement membranaire. Par ailleurs, nous avon démontré que la N-glycosylation des protéines de l’ectodomaine est indispensable à l’oligomérisation et au tri apical de GPI-APS. Nos données indiquent qu’il existe au moins deux mécanismes, l’un dépendant du taux de cholestérol et l’autre de la N-glycosylation, qui déterminent l’oligomérisation dans l'appareil de Golgi et le tri des GPI-APs vers la membrane apicale. / 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.

A incidência de infecções por fungos oportunistas na população de pacientes imunocomprometidos tem aumentado nos últimos anos, e estas são, principalmente, causadas por Candida albicans. Este patógeno oportunista pode crescer em diferentes formas, variando de levedura, pseudohifa e hifa e essa transição morfológica está associada com a virulência. Na transição de levedura para hifa, genes hifa-específicos são expressos e muitos deles codificam proteínas que possuem uma molécula de GPI (glicosilfosfatilinositol), mas a maioria (66%) das proteínas preditas ancoradas por GPI (PGAs), ainda possuem função desconhecida. Portanto, o objetivo deste trabalho, foi iniciar a caracterização funcional dos genes PGA13 e PGA58 através de análises in silico, da expressão diferencial dos RNAm sob condições ambientais diversas, e nos mutantes nulos TUP1 e EFG1, e a obtenção e caracterização fenotípica dos mutantes de PGA13: homozigoto nulo e de superexpressão. Os estudos realizados in silico indicam que PGA13 e PGA58 são reguladas pelos fatores transcricionais Efg1, Tec1 e Nrg1 e que possuem sítios potenciais de glicosilação. A análise transcricional desses genes mostra que ambos são regulados por Tup1 e que respondem aos estímulos NaCl (Cloreto de sódio), H2O2 (Peróxido de hidrogênio), etanol, cafeína, HCl (Ácido Clorídrico) e às condições hipo e hiperosmótica. Os mutantes nulos e de superexpressão de PGA13 e PGA58 foram obtidos e as linhagens que perderam o gene PGA13 tiveram um padrão de crescimento da colônia diferente da linhagem CAI-4 e foram mais sensíveis aos compostos SDS, Higromicina B e pH ácido, enquanto o mutante que superexpressava foi mais resistente. Estes resultados sugerem que Pga13 deve ter papel importante na parede celular, visto que a falta dela ocasionou maior sensibilidade a compostos que agridem a parede celular. / The incidence of opportunistic fungal infections in immunocompromised patients has increased in the last years. Candida albicans is the most commonly isolate in immunocompromised patients. C. albicans may grow in distinct morphologies: yeast, pseudohyphal and true hyphal. The switch between yeast and filamentous forms is strongly associated with virulence. During the transition, hyphal-specific genes are expressed and many of these genes encode glycophosphatidylinositol (GPI)- anchored proteins. The majority (66%) of predited GPI proteins has unknown functions. The purpose of this work was to functionally characterize the novel PGA13 and PGA58 genes. To accomplish this task, we have made in silico promoter analysis, mRNA differential expression analysis under some environmental conditions. We have also verified whether those genes are regulated by Tup1 and Efg1 transcriptional regulators. We have knocked out PGA13 and have done phenotypic analysis of the resulting PGA13 null mutant strain and a Pga13 overexpressing strain. The in silico promoter analysis indicates that PGA13 and PGA58 may be regulated by the transcriptional factors Efg1, Tec1 and Nrg1. PGA13 and PGA58 aminoacid sequence analysis revealed that they have potential glycosylation sites. The transcriptional analysis has shown that both genes are regulated by the morphogenesis negative regulator Tup1. PGA13 and PGA58 genes have differential expression in response to salt, hydrogen peroxide, ethanol, caffein, and acid stresses, and also to hypo and hyperosmotic shocks. The phenotypic analysis shows that the pga13/pga13 mutant was more sensitive to SDS, Hygromycin B and acid pH in a plate dilution sensitivity test. Interestingly, the overexpressing mutant strain was more resistant to the same compounds. Taken together these results suggest that PGA13 may have an important role in the cell wall since its absence leads to higher sensitivity to compounds that affect this organelle.

The key objective of the diploma thesis is to evaluate tourism competitiveness in the European Union -- particularly which competitiveness rankings are related to tourism sector and what position the EU and its member states represent within them. Also, there is politics of the EU analysed, key documents are mentioned and future opportunities for tourism development are outlined in the diploma thesis. In conclusion, particular project is analysed. The project is an example of above mentioned competitiveness support in tourism sector of the EU.

A coleta de hemocomponentes por equipamentos de aférese tem aumentado muito nos últimos anos, sendo considerado um avanço na medicina transfusional, pois possibilita a retirada de um ou mais componentes de um doador único resultando em um hemocomponente padronizado e de alta qualidade. No entanto, os intervalos entre as doações de plaquetaférese em geral são curtos, podendo haver perda de células a cada doação e potencial desregulação do sistema hematopoiético. Pode ocorrer ainda um possível efeito patogênico após passagem das células pelo equipamento de aférese e ativação neutrofílica. Diante disso, há a preocupação se isso acarretaria riscos à saúde do doador em longo prazo. Objetivo: O objetivo deste estudo foi avaliar a perda da expressão de proteínas ancoradas ao glicosilfosfatidilinositol (GPI), presença de clone HPN (hemoglobinúria paroxística noturna) e ativação de neutrófilos em doadores de plaquetaférese de repetição. Métodos: Estudo de caso controle, sendo 44 amostras de doadores de plaquetaférese de repetição e 44 doadores de sangue total controle. Foram coletadas amostras de sangue periférico, marcadas com os anticorpos monoclonais CD157, CD45, CD64, CD10 e FLAER (do inglês, Fluorescent Aerolysin, aerolisina fluorescente) e analisadas por citometria de fluxo. Para análise de ativação de neutrófilos, foram analisadas 17 amostras de doadores de plaquetaférese de repetição e 17 amostras de doadores de sangue total marcadas com CD64. Conclusão: Não foram encontradas alterações significativas na expressão das proteínas ancoradas ao GPI e na expressão de CD64 entre os doadores de plaquetaférese de repetição e os controles. Sugere-se que a doação de plaquetaférese de repetição não altera a expressão de proteínas ancoradas ao GPI, não gera clone HPN tampouco altera a expressão de CD64. Palavras-chave: plaquetaférese, GPI, HPN, ativação de neutrófilos. / The collection of hemocomponents through apheresis equipment has increased much in recent years, which is considered an advance in transfusion medicine because it enables the withdrawal of one or more components from a single donor, resulting in a standardized and high-quality hemocomponent. Nonetheless, the intervals between the plateletpheresis donations are generally short, which can cause loss of cells in each donation and potential dysregulation of the hematopoietic system. What can also happen is a possible pathogenic effect after the transit of the cells through the apheresis equipment and neutrophilic activation. In light of this situation, there is the concern about whether that brings risks to the donor’s health, in the long term. Objective: the objective of this study was to evaluate the loss in the expression of some glycosylphosphatidylinositol-anchored (GPI-anchored) proteins, the presence of PNH clone and neutrophils activation in repeated plateletpheresis donors. Methods: Case-control study using 44 samples of donors of repeated plateletpheresis and 44 samples of donors of whole-blood donors as controls. Peripheral blood samples were collected into tubes containing EDTA, marked with CD157, CD45, CD64, CD10 and FLAER monoclonal antibodies, and analyzed by flow cytometry. For the analysis of neutrophil activation, 17 samples of repeated plateletpheresis donors and 17 samples of whole-blood donors, both marked with CD64 were analyzed. Conclusion: No alteration in the expression of glycosylphosphatidylinositol-anchored proteins and in the CD64 expression was found. It is suggested that repeated plateletpheresis donation does not alter the expression of GPI-anchored proteins, does not generate PNH clone and neither alters the expression of CD64.

No description available.