As infecções relacionadas à assistência à saúde (IRAS) são um grave problema de saúde pública cujo prognóstico tem sido desfavorecido pela emergência e endemicidade de bactérias multirresistentes (MRs). Neste cenário, seguindo uma tendência mundial, no Brasil, infecções por cepas de Acinetobacter baumannii MRs produtoras de carbapenemases do tipo OXA são atualmente consideradas uma emergência clínica e epidemiológica. Na falta de alternativas terapêuticas efetivas para infecções relacionadas, este trabalho objetivou avaliar efeitos sinérgicos (utilizando checkerboard e time-kill) decorrentes da combinação de 10 antimicrobianos de diferentes classes, contra 8 cepas MRs de A. baumannii, clonalmente não relacionadas, produtoras de carbapenemases do tipo OXA-23, OXA-72, OXA-58 e OXA-143, representativas de diferentes centros hospitalares do Brasil. Como resultado, a combinação amicacina/tigeciclina apresentou atividade sinérgica (S= ΣCIF ≤ 0,5) e parcialmente sinérgica (PS= &#931CIF ;0,5-0,75) contra 4 (50%) cepas produtoras de OXA-143 ou OXA-72, e 2 cepas (25%) produtoras de OXA-23, respectivamente. Por outro lado, a combinação polimixina B/imipenem apresentou atividade S e PS contra 3 (37,5%) isolados OXA-143, OXA-23 ou OXA-72 positivos, e 1 (12,5%) isolado produtor de OXA-58, respectivamente. Já, a combinação amicacina/ampicilina-sulbactam foi S contra 2 (25%) A. baumannii OXA-143 ou OXA-23 positivos, sendo PS contra dois (25%) A. baumannii OXA-58 ou OXA-143/23 positivos. De interesse, foi o efeito S da combinação polimixina B/vancomicina, contra 2 cepas (25%) produtoras de OXA-72 ou OXA-23. Por outro lado, a combinação ampicilina-sulbactam/rifampicina apresentou atividade PS contra 6 (75%) cepas produtoras das variantes OXA-23, OXA-143, OXA-72 ou OXA-58. Da mesma forma, rifampicina combinada com polimixina B foi sinérgica para uma cepa OXA-23 (12,5%) e PS para 5/8 (62,5%) cepas produtoras de OXA-72, OXA-58, OXA-23/-OXA143 ou OXA-143. O efeito sinérgico da combinação polimixina B/imipenem foi confirmado, in vivo, no modelo murino de infecção, tanto por avaliação histopatológica como por redução das UFC/g pulmão ou baço (p ≤ 0,05). Finalmente, foi avaliada a atividade, in vitro, do lípide catiônico brometo de dioctadecildimetilamônio (DDA), individualmente e em combinação com tigeciclina. DDA possui efeito bactericida, e potencializou sinergicamente a tigeciclina contra 2 (25%) cepas OXA-143 ou OXA-23 positivas. A atividade do DDA, assim como a atividade da sua combinação com tigeciclina foram efetivas já na segunda hora de interação, como avaliado pelas curvas de morte. Em resumo, o efeito sinérgico decorrente do uso combinado de amicacina, tigeciclina, polimixina B, imipenem, rifampicina ou ampicilina/sulbactam, pode constituir uma alternativa terapêutica para o tratamento de infecções produzidas por cepas de A. baumannii MRs produtoras de oxacilinases, sendo que nanofragmentos catiônicos de bicamada do lipídeo sintético de DDA tem potencial para consolidar um produto de aplicação clínica. / Healthcare-associated infections (HAIs) are a serious public health issue, which have been related with an unfavorable prognosis due to the emergence and endemicity of multidrug-resistant (MDR) bacteria. In this scenario, following a worldwide trend, in Brazil, infections produced by MDR OXA-type carbapenemase-producing Acinetobacter baumannii are currently considered a clinical and epidemiological urgency. In the absence of effective therapeutic alternatives for related infections, this study aimed to evaluate synergistic effects (by using time-kill and checkerboard assays) achieved by the combination of 10 different classes of antimicrobial against 8 strains of MDR, clonally unrelated, A. baumannii strains producing OXA-23, OXA-72, OXA-58 and OXA-143 carbapenemases, being representatives of different medical centers in Brazil. As a result, the combination of amikacin / tigecycline showed synergistic (S = ΣFIC ≤ 0.5) and partially synergistic (PS = 0.5 to 0.75 ΣFIC) activity against 4 (50%) OXA-72 or OXA-143 producing A. baumannii strains, and two strains (25%) producing OXA-23, respectively. Moreover, the combination of polymyxin B / imipenem showed S and PS activity against 3 (37.5%) OXA-143, OXA-23 and OXA-72 positive isolates, and 1 (12.5%) OXA-58 producer, respectively. On the other hand, the combination amikacin / ampicillin-sulbactam was S against 2 (25%) OXA-143 and OXA-23 positive strains, being PS against two (25%) OXA-58- and OXA-143/23-producing A. baumannii. Of interest was the synergistic effect achieved by polymyxin B plus vancomycin against two strains (25%) producing OXA-72 and OXA-23, respectively. Furthermore, the ampicilina-sulbactam / rifampicin combination displayed a PS activity against six (75%) strains producing OXA-23, OXA-143, OXA-72 or OXA-58-type enzymes. Likewise, rifampicin combined with polymyxin B was S against 1 (25%) OXA-23-positive A. baumannii being PS to 5/8 (62.5%) strains producing OXA-72, OXA-58, OXA-23/-OXA143 or OXA-143. The synergistic effect of the combination polymyxin B / imipenem was confirmed, in vivo, in the murine model of infection, by using both histopathological studies and bacterial clearance from the lungs and spleen (CFU/g, p≤ 0.05). Finally, we evaluated the in vitro activity of the cationic lipid dioctadecyldimethylammonium bromide (DDA), alone and in combination with tigecycline. DDA display a bactericidal effect, enhancing synergistically the activity of tigecycline against 2 (25%) OXA-143 and OXA-23 positive strains, respectively. DDA activity alone and in combination with tigecycline was effective on the second hour of interaction, as evaluated by time-kill assays. In summary, the synergistic effect resulting from the combined use of amikacin, tigecycline, polymyxin B, imipenem, rifampicin or ampicillin / sulbactam, could be an alternative therapy for the treatment of infections caused by MDR A. baumannii strains producing oxacilinases. On the other hand, cationic bilayer nanofragments of DDA has potential for consolidating a product for medical application.

The above topic deals with the land use planning, the land-tenure and the deeds registration systems, applicable in former black urban and rural areas of KwaZulu Natal. These areas are divided into three categories, namely: 1. black townships on former black land (former KwaZulu townships); 2. rural or tribal land; and 3. black townships on former white land (Development Aid (DDA) townships). The writer firstly explains how the above categories of land were created in terms of the 1913 and 1936 land laws and how the administration and control of the first two categories was taken over by the former KwaZulu Legislative Assembly in 1986 whilst administration and control of the last category remained with the South African Development Trust. The writer critically analyses different pieces of legislation relating to the land system in the abovementioned categories of land. The writer further critically analyses the new land laws and their effect on the said land system. / Law / LL.M.

In this master’s thesis we focus on the basic properties of computer curves and their practical applicability. We explain how the curve can be understood in general, what are polynomial curves and their composing possibilities. Then we focus on the description of Bezier curves, especially the Bezier cubic. We discuss in more detail some of fundamental algorithms that are used for modelling these curves on computers and then we will show their practical interpretation. Then we explain non uniform rational B-spline curves and De Boor algorithm. In the end we discuss topic rasterization of segment, thick line, circle and ellipse. The aim of master’s thesis is the creation of the set of interactive applets, simulating some of the methods and algorithm we discussed in theoretical part. This applets will help facilitate understanding and will make the teaching more effective.

世界貿易組織協定之整體架構，其內容涵蓋了關稅暨貿易總協定、修正1947年關稅暨貿易總協定的瞭解書，以及規範貨品貿易、服務貿易，與智慧財產權的多邊貿易協定等。世界貿易組織協定中的附件1B，即是為規範服務貿易所訂之服務貿易總協定（General Agreement on Trade in Services，以下簡稱GATS）。由於服務貿易相關議題的起步與發展相形較晚，因此各國就世界貿易組織協定達成協議時，GATS中許多議題仍不成熟也缺乏共識，難以有具體之處理與規範。但是為了能在日後繼續處理這些未完成的議題，因此，乃在GATS條文中分別規定相關文字，要求會員在WTO成立生效後應繼續這些議題的討論與處理。 GATS中這些要求留待日後處理的內建（built-in）議題，分別規定在第6條第4項（國內規章）、第10條第1項（緊急防衛措施）、第13條第2項（政府採購）、第15條第1項（補貼）、以及第19條（特定承諾之談判）。其中，關於特定承諾談判的要求是屬於漸進式自由化的議題，其餘四項議題則是關於GATS規則之議題。由於GATS各項內建議題均頗為複雜與困難，對GATS整體架構都有一定程度之影響，其發展具有相當之重要性，從而本文期望能針對GATS規則制定的議題進行整理與分析，並在相關資料評估的基礎之上，提供若干之建議以供參考。惟鑑於時間與篇幅的考量，本文將限縮著重在緊急防衛、政府採購以及補貼等三項內建議題的討論上。 由於發展程度與個別國家的考量有所差異，已開發國家以及開發中國家對於此等議題的主張不盡相同，本文嘗試就此三項議題依序以第二章、第三章、第四章分別加以探討，透過整理、瞭解此三項談判的基本發展、重要爭議與相關之討論意見，就此三項談判的相關資料、重要爭議或討論意見予以扼要分析與適當論述，同時在目前的談判進度下，本文嘗試從開發中國家與已開發國家之不同立場為討論之出發點，並以前者立場作為我國之談判基調，並視我國實際情況作出調整，進而提出對此三項議題之評估與分析，以瞭解個別議題目前討論的進展，以及相關主張可能產生之影響。 最後，本文在前述各章的評估與分析的基礎之上，在第五章分別就此三項議題提出本文之建議，期望能提供相關研究人員以及有關當局作為參考。 / This thesis focuses on the three ongoing issues of the GATS built–in agenda - the negotiations on emergency safeguard measures, government procurement, and subsidies. These three issues were too complicated to be dealt with properly in the Uruguay Round, so the issues left were regulated in Articles X, XIII and XV of GATS, which mandates members to resolve the issues in the future. Because each country’s developments and considerations are widely different from one another, developed and developing countries did not achieve consensus on these issues. This thesis attempts to introduce and analyze the different discussions and opinions on these three issues. Besides the introductory Chapter, the thesis consists of another four parts. Chapter 2 first introduces and clarifies controversial issues on emergency safeguard measures, and then analyzes the three main proposals about ESM (emergency safeguard mechanism) and estimates the most recent status of the issue. Chapter 3 deals with government procurement under the GATS. After reviewing the history of the negotiation, attention is paid to the proposal from the European Community (hereafter “the EC”). Because the proposal involved with a fierce debate over market access, the EC proposal on a framework for rules for government procurement of services could not be accepted by most developing countries. Therefore, the issue has made little progress in the past years. Chapter 3 concludes with some commentaries on the issue. Chapter 4 turns attention to subsidies in services. Article XV of GATS mandates members to enter into negotiations with a view to develop multilateral disciplines which would avoid the distortive effects that subsidies in services may have on trade in services. The negotiation on subsidies in services, however, involves many important issues, such as the necessity to regulate subsidies in services, the definition of subsidies in services, elements of rules, etc. Although members made efforts to address these questions, nothing concrete has been achieved. The last chapter consists of conclusions and recommendations.

The above topic deals with the land use planning, the land-tenure and the deeds registration systems, applicable in former black urban and rural areas of KwaZulu Natal. These areas are divided into three categories, namely: 1. black townships on former black land (former KwaZulu townships); 2. rural or tribal land; and 3. black townships on former white land (Development Aid (DDA) townships). The writer firstly explains how the above categories of land were created in terms of the 1913 and 1936 land laws and how the administration and control of the first two categories was taken over by the former KwaZulu Legislative Assembly in 1986 whilst administration and control of the last category remained with the South African Development Trust. The writer critically analyses different pieces of legislation relating to the land system in the abovementioned categories of land. The writer further critically analyses the new land laws and their effect on the said land system. / Law / LL.M.

In this master’s thesis we focus on the basic properties of computer curves and their practical applicability. We explain how the curve can be understood in general, what are polynomial curves and their composing possibilities. Then we focus on the description of Bezier curves, especially the Bezier cubic. We discuss in more detail some of fundamental algorithms that are used for modelling these curves on computers and then we will show their practical interpretation. Then we explain non uniform rational B-spline curves and De Boor algorithm. In the end we discuss topic rasterization of segment, thick line, circle and ellipse. The aim of master’s thesis is the creation of the set of interactive applets, simulating some of the methods and algorithm we discussed in theoretical part. This applets will help facilitate understanding and will make the teaching more effective.

Proteins and their intricate network of interactions are fundamental to many molecular processes that govern life. Mass spectrometry-based structural proteomics represents a powerful set of techniques for characterizing protein structures and interactions. The last decade has witnessed a large-scale adoption in the application of these techniques toward solving a variety of biological questions. Addressing these questions has often been coincident with the further development of these techniques. Insight into the structures of individual proteins and their interactions with other proteins in a proteome-wide context has been made possible by recent developments in the relatively new field of chemical crosslinking combined with mass spectrometry. In these experiments crosslinking reagents are used to capture protein-protein interactions by forming covalent linkages between proximal amino acid residues. The crosslinked proteins are then enzymatically digested into peptides, and the covalently-coupled crosslinked peptides are identified by mass spectrometry. These identified crosslinked peptides thus provide evidence of interacting regions within or between proteins. In this dissertation the development of tools and methods that facilitate this powerful technique are described. The primary arc of this work follows the development and application of mass spectrometry-based approaches for the identification of protein crosslinks ranging from those which exist endogenously to those which are introduced synthetically. Firstly, the development of a novel strategy for comprehensive determination of naturally occurring protein crosslinks in the form of disulfide bonds is described. Secondly, the application of crosslinking reagents to create synthetic crosslinks in proteins coupled with molecular dynamics simulations is explored in order to structurally characterize the intrinsically disordered tau protein. Thirdly, improvements to a crosslinking-mass spectrometry method for defining a protein-protein interactome in a complex sample is developed. Altogether, these described approaches represent a toolset to allow researchers to access information about protein structure and interactions. / Graduate