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Schistosomen verursachen die Tropenparasitose Schistosomiasis mit 250-300 Millionen infizierten Menschen weltweit. Zur Analyse von Genfunktionen, durch die wirksame Medikamente entwickelt werden könnten, wird ein System zur Herstellung transgener Schistosomen benötigt. In dieser Arbeit wurde daher versucht, ein System zur Transfektion von Schistosoma mansoni von der transienten Expression von Reportergenen ausgehend zu entwickeln. Die Funktionalität der hergestellten Plasmide zur transienten Transfektion wurde durch die Expression der Reporter Enhanced Green Fluorescent Protein (EGFP) und beta-Galatosidase unter der Kontrolle der schistosomalen Promotoren für das Calreticulin, die 28 kDa-Glutathion-S-Transferase und das 70 kDa-Hitzeschockprotein in cos7-Zellen nachgewiesen. Für den Gentransfer in S. mansoni wurden Mikroinjektion, Elektroporation und Mikroprojektilbeschuss eingesetzt. Der Mikroprojektilbeschuss erwies sich als geeignete Methode, Plasmid-DNA zur transienten Expression von EGFP und beta-Galaktosidase in adulte und larvale Stadien zu transferieren. Da die beobachtete Reportergenexpression schwach war, wurde durch den Einsatz von Discosoma Red Fluorescent Protein (DsRed) als Reportergen versucht, das Transfektionsssytem zu optimieren. Außerdem wurden die potentiellen Promotoren für das Cathepsin D und für ein Aktin durch inverse PCR aus genomischer DNA isoliert. Die Funktionalität dieser Promotoren wurde in cos7- bzw. HeLa-Zellen nachgewiesen, eine verbesserte Expression in S. mansoni dagegen wurde mit diesen zwei Promotoren und unter Einsatz von DsRed nicht erreicht. Da S. mansoni sich in vitro nicht kultivieren lässt, muss für die Etablierung eines Transfektionssystems ein in seinen Keimzellen transgenes Stadium in den Lebenszyklus eingeschleust werden. Durch Mikroprojektilbeschuss transfizierte Mirazidien (freilebende Stadien des Parasiten) infizierten Zwischenwirtsschnecken und entwickelten sich zu transgenen Sporozysten, was durch die Transkription von EGFP nachgewiesen wurde. Für eine stabile Integration in das Genom werden unter anderem mobile genetische Elemente eingesetzt. Boudicca ist ein endogenes long terminal repeat (LTR) Retroelement. In dieser Arbeit wurde gezeigt, dass es in adulten Würmern, Sporozysten und Zerkarien transkribiert wird. Das LTR kann in vitro als Promotor zur Expression von EGFP eingesetzt werden. Außerdem wurde eine transkribierte Kopie kloniert und analysiert. Die in dieser Arbeit gewonnen Ergebnisse weisen darauf hin, dass Boudicca als Vektor zur stabilen Transfektion von S. mansoni eingesetzt werden kann. / Schistosomiasis caused by schistosomes affects about 250-300 million people world wide. The establishment of a transgenesis system for schistosomes is a pre-requisite to the identification of new drug targets and to the analysis of gene regulation and will add to our knowledge of parasite physiology and development. In this study, plasmids expressing Enhanced Green Fluorescent Protein (EGFP) and beta-galactosidase driven by the schistosomal promoters of the calreticulin, the 28kDa-glutathione-S-transferase and the 70kDa-heatshock protein were cloned and successfully tested in cos7-cells. Microinjection, electroporation and particle bombardment were used to introduce plasmid DNA into Schistosoma mansoni. Adult and larval stages of S. mansoni subjected to particle bombardment transiently expressed the reporter genes, although the observed transfection rates were very low. To optimize the transfection system Discosoma Red Fluorescent Protein (DsRed) was used as a reporter gene. Furthermore, the schistosomal promoters of the aspartic protease cathepsin D and a cytoplasmatic actin gene were isolated from genomic DNA using inverse PCR. The isolated promoters were able to drive EGFP and DsRed expression in cos7- or HeLa-cells, but improved expression could not be observed in S. mansoni. Due to the complexity of the life cycle S. mansoni cannot be cultured in vitro. In order to develop a transgenesis system for schistosomes, the life cycle must consequently be completed by genetically modified parasites. In this study it was shown that the free-living and mobile miracidium that infects the intermediate host snail could be transformed by particle bombardment and reintroduced in the life cycle using the natural path of infection. Development into transgenic sporocysts was shown through the isolation of EGFP transcripts from intermediate host snails. Mobile genetic elements are used as molecular tools to achieve stable integration of a foreign gene into a genome. Boudicca is an endogeneous long-terminal repeat (LTR) transposon in the schistosomal genome. In this study it was shown that it is transcribed in adult worms, sporocysts and cercariae. The LTR drives EGFP expression in cell cultures. Furthermore, a Boudicca transcript was cloned and analyzed by sequencing. The results suggest that Boudicca can be used as a molecular tool for stable integration of transgenes into the schistosomal genome.

Made available in DSpace on 2016-04-26T18:18:34Z (GMT). No. of bitstreams: 1 Tarcisio de Sa Cardoso.pdf: 5324585 bytes, checksum: 0a2380896645c26954804659ea1cba01 (MD5) Previous issue date: 2010-05-14 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / This research seeks to understand the mechanisms of non-representational contemporary graphic design. The expression "non-representative" is intended to connote some kind of sign that is characterized by polysemy. In print, this aspect has insisted to emerge in many different manifestations. Despite its recurrence, this type of sign has not received the deserved attention by the scientific community, especially in regarding to the theory of signs an instrument that seems privileged to explain the functions of language presented there. Called unreadable and dysfunctional, the graphic design of David Carson, Neville Brody, Edward Fella, April Greiman and many others are still currently subject of contents. To be more specific, we may say that all post-ulmian vanguard from around the world, questions the rationality of an area so close to the arts, and represents a kind of counterpoint to the canonical approach, which from the Bauhaus and constructivism tries to articulate an exclusivist disciplinarity to the design. This objection became important especially in Europe (HOLLIS, 2001) and in the USA (MCCOY, 1990), in which new wave movement has its most characteristic representation. One of the theorists that works with the evolution of the modernist slogan form follows function, Bürdek presents the problem of function in a new outlook not anymore as an exclusively practical function, but mainly what he called a sign function. According to him, these are the most important functions for design (BÜRDEK, 2002). From this point of view, we intend to answer how these sign functions are articulated in visuality of post-modern design from the 1990s, to build communication processes that dialogue with the culture of information, or the culture in which the sign function overcome the function of use (BAUDRILLARD, 1995). Differently from the structuralist conception of the sign in Baudrillard it means by sign function, in this study, the processes that substantiate the sign according to Peirce's semiotics (on aspects of quality, existence and regularity). From this perspective, we will intend a deeping in trichotomies of visual language as presented in matrix theory of language-thought (SANTAELLA, 2005). These trichotomies will be essential to study the controversial expressions of contemporary graphic design, in order to identify what it is innovative, which is commonplace, which transcends the current publishing paradigm and what is considered, even today, polemic by the scientific community / Esta pesquisa busca entender os mecanismos não-representativos do projeto gráfico editorial contemporâneo. O neologismo ―não-representativo‖ pretende conotar certo tipo de signo que se caracteriza pela polissemia. Na mídia impressa, esse aspecto tem insistido em despontar sob as mais variadas manifestações. Apesar de sua recorrência, esse tipo de signo não tem recebido a devida atenção por parte da comunidade científica, especialmente no que diz respeito à teoria dos signos instrumento que nos parece privilegiado para explicar as funções de linguagem ali presentes. Taxados de ilegíveis e disfuncionais, os projetos gráficos de David Carson, Neville Brody, Edward Fella, April Greiman e tantos outros são, ainda hoje, motivo de discórdia. Para ser mais específico, pode-se dizer que toda a vertente vanguardista pós-ulmiana de diversas partes do mundo questiona a racionalidade de uma área tão próxima das artes e representa uma espécie de contraponto à abordagem canônica, que desde o construtivismo e a Bauhaus tenta sistematizar uma disciplinaridade exclusivista para o design. Este contraponto ganha voz especialmente na Europa (HOLLIS, 2001) e nos EUA (MCCOY, 1990), em cujo movimento new wave tem sua representação mais característica. Um dos teóricos que trabalha com a evolução do lema modernista form follows function, Bürdek apresenta o problema da função sob um novo prisma, não mais como função exclusivamente prática, mas, principalmente, naquilo que denominou funções de signo. Segundo o autor, estas são as funções mais importantes para o design (BÜRDEK, 2002). A partir disto, busca-se responder como estas funções de signo se articulam na visualidade do projeto pós-moderno, a partir da década de 1990, para a construção de processos comunicativos que dialogam com a cultura da informação, ou com a cultura em que a função de signo supera a função de uso (BAUDRILLARD, 1995). Diferentemente da concepção estruturalista do signo em Baudrillard, entende-se por função de signo, neste estudo, os processos que fundamentam o signo segundo a semiótica peirceana (sob aspectos de qualidade, existência e regularidade). A partir dessa perspectiva, buscar-se-á um aprofundamento nas tricotomias da linguagem visual tal como apresentadas na teoria das matrizes da linguagem-pensamento (SANTAELLA, 2005). Estas tricotomias serão fundamentais para estudar as controversas expressões do design gráfico contemporâneo, com o objetivo de identificar o que nela é inovador, o que é lugar-comum, o que transcende o paradigma editorial vigente e o que nela é considerado, ainda hoje, polêmico pela comunidade científica

Police Practice / D. Litt. et Phil. (Police Practice)

Die Habilitation befasst sich mit neuen Verfahren und Anwendungen des Thermischen Spritzens, beleuchtet anhand einer internationalen Umfrage den Forschungsbedarf der Branche und liefert zahlreiche Lösungen, die im Rahmen von grundlagenorientierten und industrienahen Forschungsvorhaben erarbeitet wurden. Der Fokus der Arbeit liegt auf der Verbesserung individueller Arbeitsschritte der Prozesskette des Thermischen Spritzens, wobei Ergebnisse aus den Bereichen Werkstoffentwicklung, Prozessoptimierung, Qualitätssicherung und Nachbearbeitung zusammengeführt und durch konkrete Anwendungsbeispiele hinterlegt werden. Im Detail werden die Aspekte Herstellung leistungsfähiger und preiswerter Spritzzusätze (Wasserverdüsung von Metallpulvern, Hochenergiekugelmahlen, Agglomerieren und Sintern sowie Fülldrahtherstellung), die Verbesserung der Prozessführung (numerisch optimierte Spritzbrenner und automatisierbare Online-Prozessdiagnostikmethoden) und die Steigerung der Leistungsfähigkeit der Beschichtungen durch mechanische Nachbearbeitung sowie Versieglung behandelt. Anwendungsbezogen werden das Beschichten von Hochleistungspolymeren und CFC-Leichtbaustrukturen untersucht. Um Anknüpfungspunkte für weiterführende Forschungsarbeiten zu schaffen, schließt die Arbeit mit der Darstellung von Entwicklungstrends und zeigt Arbeitsgebiete auf, die perspektivisch von thermisch gespritzten Schichten profitieren können.

Ce mémoire étudie le phénomène de wedge politics sous un angle communicationnel, en proposant d’identifier et décrire les principales pratiques rhétoriques associées au déploiement d’une stratégie de wedge politics par les nombreux acteurs du débat public sur le projet de loi C-391, intitulé Loi modifiant le Code criminel et la Loi sur les armes à feu (abrogation du registre des armes d’épaule). La posture rhétorique que nous adoptons se traduit par une démarche méthodologique et des analyses imbriquées en quatre étapes: 1) l’élaboration d’une mise en perspective historique relativement élargie du débat public entourant le projet de loi C-391, 2) la recension des principaux acteurs et des discours qu’ils ont produits à un moment fort de ce débat, entre mai 2009 et mai 2011, 3) une première analyse et description générale de la dynamique rhétorique entre les acteurs du débat pendant cette période, et enfin, 4) une analyse systématique des discours échangés entre le 1er août 2010 et le 22 septembre 2010 nous permettant d’identifier et de décrire les principales pratiques rhétoriques employées par les acteurs. Les dix pratiques que nous avons relevées sont: l’appel à l’action, le scapegoating, le ciblage, la personnalisation du débat, le blâme, la dérision, l’attribution d’intentions malveillantes, la menace de représailles de la part des électeurs, l’exploitation des clivages et le contraste. En conclusion, nous discutons en quoi ces pratiques rhétoriques peuvent contribuer à l’atteinte des objectifs d’une stratégie de wedge politics. / This thesis examines the wedge politics phenomenon from a communicational standpoint by identifying and describing the main rhetorical practices associated with the deployment of a wedge politics strategy during the public debate surrounding Bill C-391, entitled An Act to amend the Criminal Code and the Firearms Act (repeal of long-gun registry). Our rhetorical approach entails a methodological framework and analyses deployed in four stages: 1) the establishment of a relatively wide ranging historical perspective on the public debate surrounding Bill C-391, 2) a survey of the main actors and the speeches they exchanged at a key moment of the debate, between May 2009 and May 2011, 3) a preliminary analysis and general description of the rhetorical dynamics among the actors during that time, and finally, 4) a systematic analysis of the exchanges that took place between August 1st 2010 and September 22nd 2010 in order to identify and describe the key rhetorical practices employed by the actors. The ten practices we have identified are: call to action, scapegoating, targeting, personalisation of the debate, blame, mockery, attribution of malicious intent, evoking the threat of retribution on the part of the electorate, exploiting cleavages and contrast. We conclude by discussing how these rhetorical practices contribute to the general objectives of a wedge politics strategy.

This master thesis deals with design and realization of multistage electromagnetic accelerating device (gun) and possibilities to increase its effectivity. This work contains a simulation of a magnetic circuit and designs of each components, inclusive of control circuits. Reached effectivity will be measured by appropriate methods.

The subject of my diploma thesis is sports centre. There are 3 establishments. Main establishment is shooting range. Others are gun store and café. Floor 1S is almost completely under terrain and is designed with cast-in-place concrete. Ceiling above floor 1S is designed with cast-in-place concrete slab. Floor 1NP is designed with aerated concrete blocks with ventilated facade. Ceiling above 1NP is designed with prestressed concrete floor slabs. Roof was designed as flat roof with growing layer. The drawings were processed in a computer program ArchiCad.

Die Habilitation befasst sich mit neuen Verfahren und Anwendungen des Thermischen Spritzens, beleuchtet anhand einer internationalen Umfrage den Forschungsbedarf der Branche und liefert zahlreiche Lösungen, die im Rahmen von grundlagenorientierten und industrienahen Forschungsvorhaben erarbeitet wurden. Der Fokus der Arbeit liegt auf der Verbesserung individueller Arbeitsschritte der Prozesskette des Thermischen Spritzens, wobei Ergebnisse aus den Bereichen Werkstoffentwicklung, Prozessoptimierung, Qualitätssicherung und Nachbearbeitung zusammengeführt und durch konkrete Anwendungsbeispiele hinterlegt werden. Im Detail werden die Aspekte Herstellung leistungsfähiger und preiswerter Spritzzusätze (Wasserverdüsung von Metallpulvern, Hochenergiekugelmahlen, Agglomerieren und Sintern sowie Fülldrahtherstellung), die Verbesserung der Prozessführung (numerisch optimierte Spritzbrenner und automatisierbare Online-Prozessdiagnostikmethoden) und die Steigerung der Leistungsfähigkeit der Beschichtungen durch mechanische Nachbearbeitung sowie Versieglung behandelt. Anwendungsbezogen werden das Beschichten von Hochleistungspolymeren und CFC-Leichtbaustrukturen untersucht. Um Anknüpfungspunkte für weiterführende Forschungsarbeiten zu schaffen, schließt die Arbeit mit der Darstellung von Entwicklungstrends und zeigt Arbeitsgebiete auf, die perspektivisch von thermisch gespritzten Schichten profitieren können.

Title: Effect of standing shooting position on precision of repeated shots of Hard Task Rangers. Objectives: The aim of this work is to find out in which standing stance will be the most precise shots with double tap shots. Methods: In this thesis was used descriptive analysis when collecting data about standing shooting stance problematic. Mainly through studying of literature and lecturing videos on this subject. Testing was used in the actual collecting data about shooting performance. The measurement was used to collect data on the weight, height and stability of the examined shooters. Data analysis was performed when evaluating individual shooting performance. Results: It has been found out, based on data from a group of twelve shooters from Hard Task Rangers, that even though the standing position is the furthers part of the shooting pyramid, it still provides a solid foundation for all subsequent parts of the shooters performance. Furthermore, it was found that the best standing position for firing repeated shots (double tap) was the stance called Quality universal stance. This thesis is probably the first literary study to analyze in more detail the standing shooting stance in its various forms when comparing individual attitudes and its advantages in dynamic and defensive shooting....