Thesis (Ph.D.) - University of St Andrews, June 2008.

L’agent pathogène responsable de la tuberculose, Mycobacterium tuberculosis, utilise principalement les lipides comme source d’énergie lors du processus infectieux. Cela suggère que les enzymes lipolytiques (ELs) sont des facteurs d’une importance cruciale, qui jouent un rôle essentiel pour la survie et la persistance du bacille au sein des granulomes tuberculeux. Dans ce contexte, nous nous sommes intéressés au rôle physiologique de la protéine LipY (Rv3097c), une TAG-hydrolase apparentée à la lipase Hormono-sensible humaine. En utilisant un modèle expérimental de « foamy » macrophages, nous avons démontré que cette protéine sécrétée par le système de sécrétion ESX-5 était responsable de l’hydrolyse des triacylglycerols (TAG) de l’hôte dans la lumière phagosomale. Dans une seconde étude, nous avons développé un modèle expérimental robuste et réversible basé sur la biodisponibilité en azote et en carbone, deux facteurs essentiels qui gouvernent la formation et l’hydrolyse de TAG sous la forme d’inclusions lipidiques intracytoplasmiques (ILI) chez les mycobactéries. La simplicité d’utilisation de ce modèle s’avère être un excellent système biologique permettant d’étudier les propriétés phénotypiques des bacilles riches en ILI. L’ensemble des résultats obtenus au cours de ces travaux de thèse confirme que certaines ELs sont directement impliquées dans la virulence et/ou la survie des mycobactéries pathogènes. De plus, une très grande partie de ce travail ouvre de nouvelles perspectives sur la caractérisation et l’inhibition d’acteurs protéiques essentiels au métabolisme des lipides in vivo, offrant ainsi de nouvelles opportunités pour lutter contre M. tuberculosis. / Mycobacterium tuberculosis, the pathogenic agent responsible for tuberculosis (TB), mainly uses lipids as a source of energy during the infectious process. This suggests that lipolytic enzymes (LEs) are crucial metabolic agents that play a critical role in the survival and persistence of the bacillus within TB granulomas. In this context, we investigated the physiological role of the LipY protein (Rv3097c), a TAG-hydrolase related to the human Hormono-sensitive lipase. Using an experimental model of "foamy" macrophages, we demonstrated that this protein which is secreted by the ESX-5 secretion system, was responsible for the hydrolysis of host-derived triacylglycerols (TAGs) within the phagosomal compartment. In a second study, we developed a robust and reversible experimental model based on the bioavailability of nitrogen and carbon, two essential factors that govern the formation and hydrolysis of TAGs in the form of intracytoplasmic lipid inclusions (ILI) in mycobacteria. The simplicity of this model proves to be an excellent biological system for studying the phenotypic properties of ILI-rich bacilli. All the results obtained during this thesis confirm that some LEs are directly involved in the virulence and/or survival processes of pathogenic mycobacteria. In addition, we truly believe that this work opens up new perspectives on the characterization and inhibition of protein, that are essential actors for lipid metabolism in vivo, thus offering new opportunities to better control M. tuberculosis.

Influenza is a population health issue in Canada, with an annual infection rate of 10-25% of the population. The purpose of this thesis was to analyze influenza-like illnesses (ILI) from 2004-05 through to 2008-09, both spatially and temporally, throughout the province of Manitoba. The analysis used a framework specific to spatial analysis, and incorporated the principles of population health and ecological frameworks. The underlying objectives of the research were to better understand the patterns of ILI diagnoses as well as the characteristics of those diagnosed. Maps were created to show the results from various perspectives and negative binomial regression analysis was used to test which, if any, of the chosen variables were significant. Based on this research, one could conclude that although clusters of ILI do exist in Manitoba, a clear relationship does not exist between the determinants of health and ILI.

Influenza is a population health issue in Canada, with an annual infection rate of 10-25% of the population. The purpose of this thesis was to analyze influenza-like illnesses (ILI) from 2004-05 through to 2008-09, both spatially and temporally, throughout the province of Manitoba. The analysis used a framework specific to spatial analysis, and incorporated the principles of population health and ecological frameworks. The underlying objectives of the research were to better understand the patterns of ILI diagnoses as well as the characteristics of those diagnosed. Maps were created to show the results from various perspectives and negative binomial regression analysis was used to test which, if any, of the chosen variables were significant. Based on this research, one could conclude that although clusters of ILI do exist in Manitoba, a clear relationship does not exist between the determinants of health and ILI.

Water is the source of life. Leonardo da Vinci, constructor of the drainage system of a swamp area north of Rome, said that “water is the blood of the soil”. Without water there is not fertile soil and without any of them there is no harvest. Sustainable Agriculture Management is all about how to develop and improve the use of different resources to make agriculture more sustainable. This study is about how to apply this concept into the study area. The Balkhash Lake is among the biggest lakes on earth, but due to unreasonable water utilization, the lake is facing the same fate as the Aral Sea. The lake belongs to the Ili Balkhash Basin IBB that is shared by two countries; Kazakhstan and China. Most of the water (around 80%), that feed the lake, come with the Ili River that is born on the Chinese side. Most of the water is used for agriculture purposes. Now the Chinese Government want to increase the irrigated area and develop more projects that require more water. If this is done without any improvement in the agriculture management, the IBB is under threat. The irrigation systems in the study area are very precarious. The farmers are isolated with almost no possibility to participate in the development process. The government is not interested in any research in water issues and the information that can be found about water consumption and cultivated area are proven to be far from reality. A difficult situation facing the above mention problematic. The agriculture area can grow but the water management in the Upper Ili Basin needs to be reviewed and improved. Just by doing small changes like changing rice cultivation for a crop with a higher water productivity the amount of water that can be saved is considerable. Or by importing virtual water from a region with a higher water productivity. But first a general recognition from the Chinese side about water shortage is necessary. The Ili Balkhash Basin can be sustainable if it is treated as a whole. The boundaries of the system need to be set around the whole basin. A partial view of the basin can be dangerous and jeopardize the subsistence of the natural environment of the area. When the carrying capacity of the basin is measured, the only way to grow is under this threshold. In this case, by improving water productivity with better agriculture management.

L'une des particularités de Mycobacterium tuberculosis, agent pathogène de la tuberculose, est sa capacité à accumuler des lipides dans son cytoplasme, ce qui favorise son entrée en dormance. Le séquençage du génome de M. tuberculosis a permis d'identifier certains gènes codant pour des enzymes lipolytiques, parmi ceux-ci : le gène codant pour la protéine Rv3097c aussi appelée LipY (composée d'un domaine PE et d'un domaine lipase relié par un Linker). Dans la première partie de ce travail de thèse, nous avons procédé à la caractérisation biochimique de LipY, mais aussi de ses formes mutantes LipY(∆PE), LipY(∆149) et LipY(∆170), et à l'étude d'inhibition des membres de la famille Lip apparentés à la lipase hormono-sensible humaine (Lip-HSL). Nous avons pu déterminer les propriétés cinétiques de l'activité lipase de LipY et de ses différentes formes mutantes.Dans la seconde partie, nous nous sommes intéressés à la compréhension du rôle des différents domaines de LipY, et du linker dans l'hydrolyse des lipides au cours de l'infection en utilisant des macrophages spumeux (macrophage riche en lipides) infectés au préalable par des souches de M. bovis BCG recombinantes. Ces résultats et les hypothèses posées durant ce travail de thèse, pourraient être appuyés par l'obtention de la structure tridimensionnelle de LipY. Pour cela, nous avons initié et procédé à la cristallogenèse de LipY. La poursuite des études d'optimisation des cristaux obtenus pourrait permettre d'aller plus en profondeur dans l'élucidation du rôle et du mécanisme d'action de LipY. / Mycobacterium tuberculosis is a pathogenic agent, responsible of the tuberculosis, which can store lipids into the cytoplasm. This accumulation allows the bacteria to enter in the dormancy phase. The sequencing of M. tuberculosis genome, allows to identify some genes coding for lipolytic enzymes, among which a gene coding for Rv3097c protein, also called LipY. (possessing PE domain linkto a lipase domain). During my PhD thesis, we first biochemically characterized LipY and its mutant forms LipY(ΔPE); LipY(Δ149) and Lip(YΔ170) and studied the inhibition of Lip family members related to the human hormone-sensible lipase (Lip-HSL). We determined the kinetic properties for the lipase activity of LipY and its mutants. In the second part, based on these previous results, we studied the role of these different domains and the linker on the hydrolysis of lipids during the infection phase, in infected foamy macrophages (lipids rich). For these studies, we used foamy macrophages infected by recombinants strains of M. bovis BCG (LipY(wt) and its mutants.These results and hypothesis can be confirmed and supported by resolving the tridimensional structure of LipY. Crystallogenesis tests allowed us to have some crystals of LipY(wt), which after optimization would allow us to have a better understanding of the role and action mechanism of LipY.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-03-21T18:23:16Z No. of bitstreams: 1 LenildaTeixeiraDaSilva_DISSERT.pdf: 9447586 bytes, checksum: d37926a108960b1f1abce78ad4149e93 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-03-27T21:40:12Z (GMT) No. of bitstreams: 1 LenildaTeixeiraDaSilva_DISSERT.pdf: 9447586 bytes, checksum: d37926a108960b1f1abce78ad4149e93 (MD5) / Made available in DSpace on 2017-03-27T21:40:12Z (GMT). No. of bitstreams: 1 LenildaTeixeiraDaSilva_DISSERT.pdf: 9447586 bytes, checksum: d37926a108960b1f1abce78ad4149e93 (MD5) Previous issue date: 2016-06-21 / A Detritivoria ? comum em peixes dulc?colas, sobretudo na regi?o neotropical, e para muitas esp?cies s?o escassas as informa??es sobre o trato digest?rio, como ? o caso da esp?cie Steindachnerina notonota, na qual, tem import?ncia ecol?gica como elo da cadeia alimentar, servindo de alimento para os peixes carn?voros, e atuando na depura??o dos ecossistemas aqu?ticos sujeitos ? polui??o org?nica. O objetivo do presente estudo consiste em analisar as adapta??es morfol?gicas do trato digest?rio, desta esp?cie end?mica da caatinga, regi?o do semi?rido brasileiro. Assim, foram utilizados 10 esp?cimes adultos (machos e f?meas), necropsiados, para an?lises macrosc?picas e microsc?picas do trato digest?rio. Todas as estruturas foram observadas no estereomicrosc?pio. Os ?rg?os foram fixados em formol 10% e submetidos ? colora??o de Hematoxilina e Eosina, e PAS - ?cido Peri?dico Schiff (rastros branquiais, ?rg?o epibranquial, es?fago, est?mago, intestino m?dio e intestino posterior) e analisados em microscopia de luz. Os rastros branquiais tamb?m foram fixados em paraformalde?do 10% para an?lise em microscopia eletr?nica de varredura. Os esp?cimes analisados possu?am comprimento total entre 6,4 a 13 cm, comprimento padr?o entre 4,8 a 10,3 cm, e peso de 3,23 a 35,92 g. As adapta??es estruturais no trato digest?rio foram: boca do tipo terminal; o particular complexo bucofar?ngeo; aus?ncia de dentes e da l?ngua; tem tr?s tipos de formatos de rastros branquiais com c?lulas mucosas e bot?es gustativos; possui o ?rg?o epibranquial com suas peculiaridades; um est?mago mec?nico com musculatura muito desenvolvida na regi?o pil?rica; e um intestino muito longo e enovelado, que tem internamente uma exclusiva prega helicoidal. Os aspectos morfol?gicos do trato digest?rio de S. notonota t?m rela??o ao h?bito alimentar especializado detrit?voro-ili?fago. / Detritivory is the common freshwater fishes, especially in the Neotropical region, and for many species are scarce information about the digestory tract, as is the case of the species Steindachnerina notonota, in which, it has ecological importance as links in the food chain, serving of feed for carnivorous fishes, and acting in the depuration of aquatic ecosystems subject to organic pollution. The objective of present study consists of analyzing the morphological adaptations of the digestory tract, of this endemic species of the caatinga, Brazilian semiarid region. Thus, 10 adult specimens were used (males and females), necropsied for macroscopic and microscopic analysis of the digestory tract. All structures were observed in the stereomicroscope. The organs were fixed in 10% formalin and submitted to hematoxylin and eosin staining, and PAS - periodic acid Schiff (gill rakers, epibranchial organ, esophagus, stomach, midgut and hindgut) and analyzed by light microscopy. The gill rakers were also fixed in 10% paraformaldehyde for analysis by scanning electron microscopy. The specimens analyzed had total length between 6,4 to 13 cm, standard length between 4,8 to 10,3 cm, and weight of 3,23 to 35,92 g. The structural adaptations in the digestory tract were: mouth of the terminal type; the particular complex bucopharyngeal; absent from tooth and tongue; it has three types of formats of gill rakers with mucous cells and taste buds; possess the epibranchial organ with its peculiarities; a mechanical stomach with highly developed musculature in the pylorus; and a very long and coiled intestine, which internally has exclusive helical fold. The morphological aspects of digestory tract of S. notonota are related to detritivore-ilyophago specialized feeding habit.

This thesis aims to revise the established history of the passage of the Second Crusade through the Byzantine Empire and Anatolia in 1147. In particular, it seeks to readdress the ill-fated advance of the army nominally headed by King Conrad III Staufen of Germany towards Ikonion, the fledging Seljuk capital of RÅ«m. The work consists of four mutually supportive parts. Part I serves to introduce the thesis, the historiographical trends of the current scholarship, and the Byzantine notion of the Latin 'barbarian', a stock, literary representation of the non-Greek other which distorts the Greek textual evidence. Part II analyses the source portrayal of particular incidents as the army marched through the Byzantine Empire, provides analyses of those events based on new approaches to interpreting the sources and a consideration of the army's logistical arrangements, and argues that the traditional historiography has been and continues to be subject to textual misrepresentation. An understanding of the topology of Anatolia is required to appreciate why the army failed to reach Ikonion. Part III therefore consists of chapters devoted to the geography of Anatolia, the form, function and the population density of the typical twelfth-century town, the country's changeable medieval geopolitical landscape, and the settlement patterns and the way of life of western Anatolia's pastoral-nomadic warriors. Part IV revisits the Latin, Syriac and Greek sources which constitute the written history of the crusade in Anatolia, analyses the concerns of the army's executive decision makers within geopolitical, logistical, topographical and tactical frameworks, and offers a reconsideration of the established location of where the army ceased to advance on Ikonion, and a new version of the circumstances which led to the decision to retreat.

Along NS trending road cut between &Ccedil / ayyolu and incek, relatively thick tuffite layer of the Han&ccedil / ili formation has been exposed. It is alternating with other lacustrine deposits such as clayey limestone, marl, siltstone and mudstone. These units unconformably overlie the Mesozoic basement limestones of Akbayir formation. The illite rich tuffite consists of glass shards tridymite, biotite, quartz and plagioclase. It has a persistent areal extend and forms the foundation of the residential buildings at several localities. In this thesis, it is aimed to assess the engineering geological properties of the tuffite layer and to investigate its suitability as a foundation material. In order to accomplish this task, field studies and laboratory tests were conducted. Field studies involve detailed site geological observation. Laboratory studies include tests to investigate mechanical, physical and mineralogical properties of the tuffite. v Analyses revealed very weak nature of tuffite with strength values ranging between 0.1 MPa and 1.00 MPa. The tuffite is characterized with very high effective porosity (43.3%) and very low dry and saturated densities (1.19 Mg/m3 and 1.62 Mg/m3, respectively). The slake-durability tests yield medium durability. The durability index of tuffite is rated as very poor. Considering index properties and the durability assessment it is concluded that tuffite serves as a poor foundation material.

No description available.