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Die Wnt/ β-Catenin-Signaltransduktion nimmt eine exponierte Stellung in der kolorektalen Karzinogenere ein. Niclosamid ist ein Derivat der Salicylsäure, das bei Bandwurm- infektionen eingesetzt wird. Es konnte gezeigt werden, dass Niclosamid den Wnt/ β-Catenin-Signalweg moduliert. Ziel der vorliegenden Arbeit war es, den therapeutischen Einsatz des Niclosamids beim kolorektalen Karzinom zu evaluieren. Die Zellproliferation von kolorektalen Karzinomzelllinien (humane SW480 und SW620 Zellen sowie CC531 Zellen einer Ratte) und von Rattenfibroblasten wurde nach 12 und 24 Stunden Inkubation mit Niclosamid durch lichtmikroskopische Zellzahlbestimmungen beurteilt. Die Apoptoseraten wurden mit einem Zelltod-Assay ermittelt. Eine Immunfluoreszenzfärbung gab Aufschluss über das Expressionsmuster von aktivem β-Catenin. Die Promotoraktivität des LEF/ TCF-Transkriptionsfaktors wurde nach Transfektion mit TOPflash mithilfe eines Luciferase Assays analysiert. Die Genexpression von Wnt-modulierenden Faktoren (Bcl-9 und Wif1), von Komponenten des ß-Catenin- Degradationskomplexes (Axin2 und GSK 3β), von kanonischen Zielgenen (Met, MMP7 und Cyclin D1) und von c-jun als Schlüsselprotein des nicht-kanonischen Wnt/ JNK-Signalweges wurde in der RT-PCR untersucht. Auf Proteinebene wurden zur Bestätigung zusätzlich Western Blots mit Antikörpern gegen aktives β-Catenin und c-jun durchgeführt. Die Zellproliferation kolorektaler Karzinomzelllinien wurde dosisabhängig inhibiert, und Niclosamid führte zu Apoptose. Nach Inkubation mit Niclosamid kam es nicht zur Umverteilung von aktivem β-Catenin von der nukleären in die zytosolische Fraktion. Die Wnt-Promotor-Aktivität von LEF/ TCF wurde nach 12 Stunden Inkubation mit 10 und 50 μM Niclosamid jedoch signifikant gesenkt. Kanonische Wnt-Zielgene (Met, MMP7 und Cyclin D1) sowie der Koaktivator Bcl-9 wurden auf Transkriptionsebene gehemmt, während das nicht-kanonische Schlüsselprotein c-jun aktiviert wurde. Fasst man zusammen, so führt die Inkubation mit Niclosamid zu inhibitorischen Effekten auf kolorektale Karzinomzelllinien und zu einer reduzierten kanonischen Wnt-Aktivität. Diese Effekte können durch eine gestörte Formation des Triple-Komplexes aus Bcl-9, β- Catenin und LEF/ TCF und einer Aktivierung von c-jun und damit des nicht-kanonischen Wnt/ JNK-Signalweges bedingt sein. In in vivo-Untersuchungen beabsichtigen wir, in einem Tiermodell die Daten zu verifizieren und so den Einsatz des Niclosamids als Option für Patienten mit metastasiertem kolorektalem Karzinom weiterführend zu beurteilen.

Cancer is a leading cause of death worldwide. Development of new cancer drugs is increasingly costly and time-consuming. By exploiting massive amounts of biological data, computational repositioning proposes new uses for old drugs to reduce these development hurdles. A promising approach is the systematic analysis of structural data for identification of shared binding pockets and modes of action. In this thesis, I developed the Protein-Ligand Interaction Profiler (PLIP), which characterizes and indexes protein-ligand interactions to enable comparative analyses and searching in all available structures. Following, I applied PLIP to identify new treatment options in cancer: the heat shock protein Hsp27 confers resistance to drugs in cancer cells and is therefore an attractive target with a postulated drug binding site. Starting from Hsp27, I used PLIP to define an interaction profile to screen all structures from the Protein Data Bank (PDB). The top prediction was experimentally validated in vitro. It inhibits Hsp27 and significantly reduces resistance of multiple myeloma cells against the chemotherapeutic agent bortezomib. Besides computational repositioning, PLIP is used in docking, binding mode analysis, quantification of interactions and many other applications as evidenced by over 12,000 users so far. PLIP is provided to the community online and as open source.

Overall, this study enhances our understanding of the role of c-Myc activation in chromosome 11 repositioning in mouse PreB v-abl/myc cells and a possible interaction between telomeres, TRF2, and lamin A/C underlying this phenomenon. Additionally, the importance of human 17q25.3 is confirmed as a potential region involved in NSCLC tumorigenesis. A utilizationof the 3D telomeric organization profiles is demonstrated a tendency to categorize NSCLC patients into different prognostic subgroups, underscoring a potential future value of its clinical application. / February 2017

Abstract   Background:Low back pain has a reported lifetime prevalence of about 70% and tops the list of  years lived with disease in the developed countries. There is still to date areas on the mechanisms driving pain andmovement system impairments not fully understood. For some areas the  research are evident butclinically friendly methods lack. A new test for measuring joint position  sense (JPS) in the lumbar spine is tested forreliability.  Objectives:Aim of this study was to evaluate test-retest reliability on a new test measuring JPS, using two laser pointers attached to the vertebrates off L1 and S1. Design:Cross-sectional observational test-retest. Methods:82 participants, 41 with mechanical low back pain and 41 healthy controls, were tested  for repositioning error two times with 30-60minutes between tests. Movement directions tested  was; Flexion, extension, rotation right and rotation left in sitting. Intraclasscorrelation coefficient  (ICC) was used for measuring relative reliability and standard error ofmeasurement (SEM) for  absolute reliability. Results:ICC in the LBP group ranged from -0,51 – 0,94 and for the whole group -0,19 – 0,84. The SEM in the LBP group ranged from 0,1 – 2,9 (95%CI -5,6 – 6,4) and for the whole group 0,1 – 3,2 (95%CI -6,3 – 6,2). Conclusions:The test-retest reliability of this JPS test shows poor to moderate reliability. Measuring joint positioning sense by using two laser pointers attached to S1 and L1 as in this study has shown not to be reliable enough to be used in clinical tests or research and can therefore not be recommended.

O presente trabalho trata do planejamento da movimentação de contêineres vazios ao longo de um conjunto de portos, buscando o balanceamento entre as demandas e ofertas dos mesmos em todos os portos ao menor custo, e considerando as restrições de capacidade de transporte dos modais envolvidos. Para isso será adotado um modelo de fluxo em rede multi-produto para representar o sistema de movimentação de contêineres vazios e que servirá de base para o desenvolvimento de uma formulação matemática, a qual, implementada através de uma ferramenta computacional de otimização, determina os fluxos de vazios no sistema. A verificação do modelo proposto deu-se através de testes em problemas reduzidos de movimentação de vazios, assim como em um problema cujos resultados foram publicados na literatura. Os resultados sugeriram a adequabilidade e confiabilidade do modelo proposto que pode, então, ser aplicado a um problema real da empresa de navegação Hamburg Süd, tendo seus resultados comparados aos resultados fornecidos pela mesma. / This dissertation deals with the empty containers movement planning throughout a set of ports, aiming the balancing between the demands and supplies in all the ports at minimal cost, and considering the capacity constraints of the transport modes considered. A multi-commodity network flow model will be adopted to represent the empty containers movement system. This model supports the development of a mathematical formulation which, through a computational optimization tool, determines the flows of empty containers throughout the system. The verification of the proposed model was given through tests in reduced problems, as well as in a problem which results had already been published in literature. The results had suggested the adequateness and trustworthiness of the proposed model, which could, then, be applied to a real problem of the navigation company Hamburg Süd, and the results could be compared with the ones given by the company.

Devido à progressiva importância atribuída à marca e, em especial, ao seu gerenciamento, diferentes teorias e modelos de compreensão, análise e prescrição foram e vêm sendo desenvolvidos, procurando se orientar abordagens e decisões estratégicas de negócio. Embora seja possível identificar algumas diferenças conceituais e/ou estruturais, dois aspectos, na prática, estão sempre presentes na análise de marcas e podem ser considerados decisivos na compreensão e gestão dessas: identidade e posicionamento de marca. Em relação a este segundo conceito, torna-se, ao longo do tempo, mais relevante, decisivo e crucial o processo de análise e avaliação da eficácia, adequação e relevância do posicionamento, em termos organizacionais, o entendimento da condição da marca e sua relação com o mercado, assim como modelos e procedimentos de avaliação para a decisão por sua eventual modificação ou adequação ao longo do tempo: o reposicionamento. Atualmente, o exame e a decisão de um reposicionamento de marcas está presente no cotidiano de profissionais de marketing e comunicação, porém não existe consenso ou uniformidade de abordagem deste tema, resultado, entre outras razões, da limitada literatura desenvolvida acerca do assunto. Este trabalho se propõe a abordar de forma estruturada conceitual e estrategicamente condições, desafios e riscos da adoção de uma estratégia de reposicionamento de marcas, utilizando argumentação baseada em literatura disponível, somada a intervenções exploratórias de exemplos reais, e propondo uma tipificação para as decisões e estratégias de reposicionamento, assim como orientações e alternativas para a gestão de marcas. A decisão por um reposicionamento de marca envolve desafios, investimentos e riscos, sendo que, na maioria das vezes, este processo se impõe ao gestor de marca. Com o objetivo de contribuir no exame e/ou avaliação do reposicionamento de marcas, este trabalho aborda as condições, as decisões e os impactos potenciais, oferecendo uma classificação e uma estrutura de análise orientadas para operação e gestão de estratégias de reposicionamento. / Different theories and comprehension, analysis and prescription models were and are being developed associated to progressive importance of brand management. Their objectives, in general, are to offer approached-based and strategic decisions for business. However, it is possible to identify some conceptual and/or structural differences among these papers, two aspects, in practice, are always present in brand’s analysis and may be recognized as decisive to understanding and administration brands: identity and positioning. This second concept is becoming more important and crucial as time goes by, playing a fundamental role in the brand efficacy evaluation, relevancy and compatibility of positioning and, particularly, the brand condition comprehension and its relationship with the market, as evaluation procedures and decision models for brand positioning change: the repositioning. Nowadays, brand repositioning analysis and decision are part of daily activities of marketing and communication professionals, but there is not approach consensus or uniformity to deal with this concept. As matter as fact, it is result – among other reasons – of limited developed literature about this issue. That thesis proposes itself to approach structurally, conceptually and strategically the conditions, challenges and risks to the adoption of brand repositioning strategy, using argumentation based on available literature added exploratory intervention in actual cases and proposing a taxonomy for repositioning strategies and decisions and brand management orientation and alternatives. Brand repositioning decision involves challenges, investments and risks. In general, this process in not a management decision, but an external imposition as a function of realized brand performance. With the objective to contribute to analysis and evaluation of brand repositioning, this research effort focus conditions, decisions and potential impacts, offering a structure and a classification oriented to repositioning strategies an operation management.

Objetivos: O objetivo deste estudo foi o de identificar compostos seletivamente tóxicos ao carcinoma espinocelular de boca in vitro por meio do reposicionamento de fármacos. Material e Métodos: Por meio de um escaneamento baseado na viabilidade celular de 1.280 fármacos, nós selecionamos três princípios ativos (luteolin, metixene hydrochloride e nitazoxanide) letais às células de câncer de boca SCC-25 e pouco tóxicos às células de queratinócitos cutâneos imortalizados HaCaT. Os fármacos candidatos foram investigados quanto à sua dose- e tempo-resposta bem como comparados e combinados à agentes quimioterápicos padrão por meio do ensaio por colorimetria com brometo de tiazolil azul de tetrazolio (MTT). O impacto dos fármacos na motilidade do SCC-25 e do HaCaT foi verificado pelo ensaio de migração celular e seus mecanismos de ação também foram explorados por meio da verificação dos níveis das proteínas fosforiladas pelo western blotting. Todos os experimentos foram realizados em triplicata e, pelo menos, três vezes independentes. O teste t de student foi utilizado para confrontar as variáveis e nível de significância de 5% foi estabelecido para todos os testes. Resultados: O flavonoide natural luteolin exerceu citotoxicidade potente contra as células de câncer de boca in vitro, apresentando baixa toxicidade ao HaCaT e alta eficiência quando comparado a quimioterápicos como a cisplatina e o AG1478. Do ponto de vista molecular, a luteolin ativou a via de sinalização do dano do DNA e, combinada com um inibidor do Chk, apresentou efeitos potencializados. Além disso, nós demonstramos que a nitazoxanide e o metixene hydrochloride são capazes de destruir células SCC-25 porém não as HaCaT de maneira proporcional à dose e ao tempo de tratamento. As combinações entre os três fármacos hit e com a cisplatina ou o AG1478 potencializaram seus efeitos contra as células malignas. Conclusões: O presente estudo traz a luteolin, o metixene hydrochloride e a nitazoxanide como fortes candidatos a agentes terapêuticos contra o câncer de boca uma vez que estes compostos apresentam maior eficácia contra células de câncer de boca e menor toxicidade contra células não tumorais in vitro do que agentes quimioterápicos convencionais. / Objectives: Here we aimed at identifying and reposition approved drugs that could be selectively toxic towards oral squamous cell carcinoma cells. Materials and Methods: Through a cell-based drug screening of 1,280 chemical molecules, we selected 3 compounds (luteolin, metixene hydrochloride, and nitazoxanide) lethal to oral cancer SCC-25 cells, while sparing immortalized keratinocyte HaCaT cells. The drugs were then further challenged for their time- and dose-responses, as well as their comparison and combination to standard chemotherapeutic agents by colorimetric assay 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan, Thiazolyl blue formazan (MTT). The impact on SCC-25 and HaCaT motility as well as the mode of action of the drugs was then further explored by scratching assay and western blotting, respectively. All the experiments were performed in triplicated and, at least, three independent times. Students t test was performed to verify the differences among the variables and the level of significance was set at 5%. Results: The natural flavonoid luteolin was a potent cytotoxic agent against oral cancer cells in vitro, presenting low toxicity against HaCaT cells and high efficiency as compared to standard-of-care, such as cisplatin and AG1478. From a molecular standpoint, luteolin coopted the DNA-damage pathway and could be efficiently combined with Chk pharmacological inhibitor. Moreover, we demonstrated that nitazoxanide and metixene hydrochloride kill the SCC-25 but not the HaCaT cells in a dose- and time-dependent. The combinations among the three drugs hit and with cisplatin and AG1478 improved their effect against the malignant cells. Conclusions: Luteolin, metixene hydrochloride, and nitazoxanide emerge as strong cytotoxic and/or adjuvant therapy in oral cancer, as these compounds present higher efficiency and lower toxicity against oral cancer cells in vitro than conventional chemotherapeutic agents.

Este trabalho acata Musica ricercata (1951-53), importante obra do período húngaro de György Ligeti, sob o ponto de vista da performance musical. Estruturado em três capítulos, no primeiro investiga a maneira como o conjunto de onze peças reflete o reposicionamento artístico e político de Ligeti nos anos anteriores à sua saída de Budapeste em direção à Europa Ocidental, ocorrida em 1956, e se firma como marco de sua independência artística. No segundo capítulo, centramos a pesquisa no projeto composicional de Musica ricercata com objetivo de identificar tópicos de sua organização estrutural que seriam aprofundados por Ligeti em sua obra madura e, consequentemente, reafirmam a profunda reorientação tratada no capítulo anterior. Por fim, o terceiro capítulo aborda cada uma das onze peças do ciclo e investiga as relações de reciprocidade entre seu pensamento composicional, a notação empregada pelo compositor e, finalmente, sua execução. / The present study regards Musica ricercata (1951-53), an important work from György Ligeti\'s Hungarian period, from the perspective of musical performance. Structured in three chapters, the first one investigates how this collection of eleven pieces reflects both Ligeti\'s artistic and political positioning in the years that preceded his exit from Budapest towards Western Europe in 1956 and stands out as a turning point for his artistic independence. In chapter 2, the research focuses on Musica ricercata´s compositional project in order to identify some topics of its structural organization which would be amplified by Ligeti on his mature work and, consequently, reaffirm the deep reorientation considered in chapter 1. Finally, the third chapter discuss each one of the eleven pieces from the cycle in order to investigate reciprocity relations among his compositional thinking, the notation used by the composer and, eventually, their performance.