A leishmaniose visceral (LV) no Brasil é causada pela espécie Leishmania chagasi (L. infantum) e os cães são considerados o principal reservatório doméstico desse parasita. Portanto, o diagnóstico correto e seguro é muito importante para evitar a transmissão da doença ou o sacrifício desnecessário de cães. O objetivo deste trabalho foi avaliar o swab conjuntival (SC), um método não invasivo de amostragem, para o diagnóstico da LV canina por PCR-hibridização. Primeiramente, um teste in vitro foi delineado utilizando-se swabs contaminados com 103 até 1,0 célula de L. chagasi. Três métodos de extração de DNA foram testados: fenol-clorofórmio, kit Wizard e fervura. Em seguida, dois grupos de 23 cães soropositivos foram utilizados. Foram feitos esfregaços por SC em ambos os olhos de cada animal. A extração de DNA de SC foi realizada utilizando-se o método fenolclorofórmio no grupo 1 e a técnica de fervura no grupo 2. Além disso, sangue foi coletado de cada cão sendo que 30&#956;L foram aplicados em papel filtro (PF) e 1,0mL foi tratado para obtenção do anel leucocitário (AL). A purificação de DNA para PF e AL foi realizada da mesma forma em ambos os grupos utilizando-se kits comerciais. A análise de todas as amostras foi feita por PCR seguida de hibridização com sondas de DNA marcadas com 32P. O experimento com swabs contaminados com L. chagasi mostrou positividade na PCR para até 25, 10 e 1,0 célula pelos métodos da fervura, kit Wizard e fenol-clorofórmio respectivamente. A hibridização aumentou a sensibilidade para até 10 e 1,0 célula pelos métodos da fervura e kit Wizard respectivamente. As positividades da PCR para os cães do grupo 1 e 2 foram respectivamente: 73,9% e 52,2% (SC), 8,7% e 30,4% (AL), 8,7% e 17,4% (PF). A hibridização aumentou essas positividades para: 91,3% e 65,2% (SC), 21,7% e 34,8% (AL), 30,4% e 43,5% (PF) respectivamente. Os melhores resultados foram obtidos pela associação de SC com o método de fenol-clorofórmio para extração de DNA. O resultado final obtido desse tratamento foi estatisticamente diferente daqueles referentes ao AL e PF no grupo 1 (p<0,01). No grupo 2, o resultado final proveniente do SC associado com a técnica de fervura para o isolamento de DNA foi estatisticamente distinto apenas do resultado referente ao AL (p<0,05). Concluiu-se que a combinação de SC com a extração de DNA por fenol-clorofórmio é uma ferramenta valiosa para o diagnóstico da LV canina e pode ser recomendada para o diagnóstico em cães sintomáticos. / The visceral leishmaniasis (VL) in Brazil is caused by Leishmania chagasi (L. infantum) and dogs are considered to be the main domestic reservoir. Therefore the correct diagnosis is very important in order to avoid the disease transmission or unnecessary culling of dogs. The aim of this study was to evaluate the conjunctival swab (CS) as a noninvasive sampling method for the canine VL diagnosis by the PCR-hybridization procedure. Firstly, an in vitro test was carried out using cotton swabs seeded with 103 to one cell of L. chagasi. Three DNA purification techniques were tested: phenol-chloroform, Wizard kit and boiling. After that, two groups of 23 seropositive dogs were used. CS samples were obtained from both eyes of each animal. The DNA extraction from CS was performed by the phenol chloroform method in group 1 and by boiling in group 2. In addition, blood was collected from each animal so that 30&#956;L were spotted onto filter paper (FP) and 1.0mL was treated to obtain the buffy coat (BC). The DNA extraction from the BC and FP was accomplished by the same way in both groups using commercial kits. The analysis of all samples was made by PCR associated to the hybridization with 32P labeled DNA probes. The in vitro test with seeded parasites showed positivity to until 25, 10 and one cell for boiling, Wizard kit and phenol chloroform methods respectively. The hybridization step increased the sensitivity until 10 and one cell for the boiling and Wizard respectively. The PCR positivities for the dogs in groups 1 and 2 were respectively: 73.9% and 52.2% (CS), 8,7% and 30.4% (BC), 8.7% and 17.4% (FP). The hybridization step increased the positivities for: 91.3% and 65.2% (CS), 21.7% and 34.8% (BC), 30.4% and 43.5% (FP) respectively. The best frequency of positivity was obtained by the association between CS and the DNA extraction by phenol chloroform. The final result obtained from this treatment was statistically different from the BC and FP data in group 1 (p<0,05). In group 2, the result from CS associated with DNA purification by boiling was distinct only from the BC data (p<0,05). We conclude that the CS associated with the DNA extraction by phenol chloroform is a valuable tool for diagnosis of canine LV and can be recommended for diagnosis of symptomatic dogs.

Dermatófitos são fungos filamentosos com a habilidade de invadir substratos queratinizados e causar dermatofitoses em humanos e animais, penetrando profundamente apenas em hospedeiros imunocomprometidos. Trichophyton rubrum é um fungo antropofílico e cosmopolita, o mais comum agente de micoses superficiais, que usa componentes celulares como proteínas e lipídeos após uma específica regulação de sua expressão gênica governada pelo pH ambiente e sensoriamento celular. A virulência de T. rubrum é relacionada com a secreção de enzimas proteolíticas, um importante fator determinante na invasão, utilização e subseqüente disseminação através do estrato córneo. O objetivo desse estudo foi identificar através de Hibridização Subtrativa Supressiva (SSH) genes de T. rubrum preferencialmente expressos durante o crescimento na presença de queratina e lipídeos, quando T. rubrum degrada fontes de carbono tipicamente encontradas em células epidérmicas. Inicialmente, nós avaliamos as mudanças no pH extracelular durante seu crescimento em queratina e lipídeo (depois de 6, 12, 24, 48, 72 h e 7 dias) em pH inicial 5,0, onde foi observado um gradual aumento do pH basal sob ambas as condições de teste, comparado com a condição glicose (controle). Também identificamos 576 transcritos de T. rubrum diferencialmente expressos por SSH usando conídios cultivados por 72 h em queratina como teste e em glicose como controle. Os genes de T. rubrum ativados codificam proteínas putativas que foram validadas por cDNA dot-blot e northern blot, mostrando similaridade com proteínas fúngicas envolvidas no metabolismo básico, crescimento e virulência, p. ex., transportadores ABC-MDR, MFS e ATPase de cobre, permease, NIMA interactive protein, poliproteína Gag-Pol, fatores de virulência subtilisinas serino-proteases (Sub 3 e 5) e metaloproteases (Mep 3 e 4) e Hsp30. Adicionalmente, entre os 762 clones obtidos na biblioteca da condição lipídeo (72 h), 80 transcritos superexpressos foram confirmados por cDNA dot-blot, revelando 14 unigenes similares à proteínas de vários organismos patogênicos, como glicoproteína 43 kD, transportador MDR, proteína G, quitina sintase e serino/treonina fosfatase. Transcritos do gene TruMDR2, codificador de um transportador ABC, foi isolado tanto na presença de queratina quanto em lipídeo, e a análise da linhagem mutante TruMDR2 de T. rubrum mostrou uma redução na atividade infectante, caracterizada pelo baixo crescimento em unhas humanas comparada com o tipo selvagem. A alta expressão de transportadores por T. rubrum em condições que mimetizam a infecção e a redução na virulência de TruMDR2 durante o modelo in vitro sugerem que transportadores estão envolvidos na patogenicidade de T. rubrum. Outro linhagem mutante (pacC-1) com um nocaute no gene pacC que codifica um fator de transcrição regulado pelo pH local, mostrou a expressão de proteases (Sub 3 e 5 e Mep 4) diminuída após o crescimento em queratina em comparação com o tipo selvagem em análises de northern blots. Essas proteases tem uma atividade ótima em pH alcalino, e nossos resultados indicam uma regulação defectiva do gene pacC de T. rubrum na ativação de proteases. Em conclusão, através do uso de SSH foi possível identificar genes de T. rubrum ativados após tratamentos específicos, o que sugere a importância dos mesmos na interação dermatófito-hospedeiro, instalação e manutenção da doença. Esses resultados disponibilizam novos dados sobre T. rubrum que levarão a um melhor entendimento dos mecanismos moleculares envolvidos no crescimento, metabolismo geral e patogenicidade, e também auxiliar na identificação de novos e efetivos alvos de drogas para dermatófitos. / Dermatophytes are a group of fungi filamentous that have the ability to invade keratinized substrates, causing dermatophytosis in humans and animals and only penetrate deeper if the host is immunocompromised. Trichophyton rubrum is an anthropophilic and cosmopolitan fungi, the most common agent of superficial mycoses, which uses cell components such as proteins and lipids after a specific regulation of its gene expression governed by pH environment and sensing cell. The virulence T. rubrum is related to secretion of proteolytic enzymes, an important factor determinant in the invasion, utilization and subsequently dissemination through the stratum corneum. The aim of this study was to indentify by Suppression Subtractive Hybridization (SSH) T. rubrum genes preferentially expressed during growth in the presence of keratin and lipids, upregulated when this fungus degrades carbon source typically found at epidemic cells. Initially, this work evaluated the changes in the extracellular pH during its growth in keratin and lipid (after 6, 12, 24, 48, 72 h and 7 days) at initial pH 5.0, where we observed a gradual increase of basal pH under both tests when compared to glucose condition (control). Also, we identified 576 T. rubrum transcripts differentially expressed by SSH using conidia cultivated for 72 h in keratin as tester, and in glucose as driver. The T. rubrum genes upregulated encode putative proteins that were validated by cDNA dot-blot and northern blot, showing similarity to fungi proteins involved in basic metabolism, growth and virulence, i.e., transporters ABC-MDR, MFS and ATPase of copper, permease, NIMA interactive protein, Gag-Pol polyprotein, virulence factors serine-protease subtilisins (Sub 3 and 5) and metalloproteases (Mep 3 and 4) and Hsp30. Additionally, among the 762 clones obtained in a library of lipid condition (72 h), 80 over-expressed transcripts were confirmed by cDNA dot-blot, revealing 14 unigenes similar to proteins of several pathogenic organisms, like glicoprotein 43 kD, MDR transporters, G protein, chitin synthase and serine/threonine-protein phosphatase. Transcripts of TruMDR2 gene, encoding an ABC transporter in T. rubrum, were isolated in the presence of keratin and lipid, and the examination of TruMDR2 mutant T. rubrum showed a reduction in infecting activity, characterized by low growth in human nails compared to wild-strain. The high expression of transporter by T. rubrum in conditions that mimetize the infection and the virulence reduction of TruMDR2 in an in vitro model suggests that transporters are involved in T. rubrum pathogenicity. Another mutant, pacC-1 with a knockout in PacC gene that encodes a transcription factor regulated by local pH, showed the expression of proteases (Sub 3, Sub 5 and Mep 4) decreased after growth in keratin (72 h) in comparison to wild-strain in northern blot analyzes. These proteases have an optimal activity in alkaline pH, and our results indicate a defective regulation of T. rubrum pacC gene in the activation of proteases. In conclusion, by means of SSH to identify genes upregulated in T. rubrum after specific treatments, their importance in the dermatophyte-host interaction, installation and maintenance in the disease is suggested. These results provide new insights about T. rubrum that will contribute to a better understanding of molecular mechanisms about the growth, metabolism and pathogenicity, and may also aid in the identification of novel effective drug targets for dermathophytes.

Sistemas poliméricos para a liberação de fármacos associados à terapêutica periodontal convencional têm sido desenvolvidos e aplicados com algumas vantagens em comparação ao debridamento mecânico isolado. Um dispositivo de liberação lenta para uso em bolsas periodontais contendo metronidazol como princípio ativo, foi desenvolvido e testado quanto à sua eficácia no tratamento de pacientes com periodontite crônica. Dezesseis pacientes foram selecionados e aleatoriamente alocados aos seguintes tratamentos com 8 indivíduos por grupo: 1) raspagem e alisamento radicular (controle ativo) ou 2) raspagem e alisamento radicular + aplicação de gel contendo metronidazol a 15% (experimental). Os efeitos dos tratamentos foram avaliados pelo acompanhamento longitudinal de parâmetros clínicos (profundidade do sulco gengival à sondagem e nível clínico de inserção) e microbiológicos (análise da microbiota subgengival pela técnica da hibridização DNA-DNA Checkerboard). O monitoramento das concentrações de metronidazol no fluido gengival colhido dos pacientes que receberem o gel foi realizado pela cromatografia líquida de alta eficiência. Os resultados da análise das concentrações de metronidazol na bolsa periodontal demonstraram que o gel só pôde ser detectado em concentrações efetivas até uma hora depois da aplicação. Em relação à análise microbiológica, foi observada diferença significante entre os grupos, nos três tempos de avaliação (antes, 7 e 30 dias depois do tratamento). A análise intragrupos indicou redução significante na contagem bacteriana somente para o grupo tratado com o gel de metronidazol, após 7 dias do tratamento. Embora tenha sido detectada diferença entre os grupos nenhum deles foi capaz de eliminar ou reduzir com significância as espécies periodontopatogênicas. No que se refere aos parâmetros clínicos, ambas as terapias foram eficazes em promover a redução na profundidade à sondagem e o ganho no nível de inserção, sem benefício significante para o emprego do gel de metronidazol. / Polymeric systems for drug delivery have been developed and implemented with several advantages compared to conventional therapy. A slow-release device for use in periodontal pockets containing metronidazole as active principle, was developed and tested for its efficacy in treating patients with chronic periodontitis. Sixteen patients were randomly allocated to treatments with eight subjects per group: 1) scaling and root planing (active control) or 2) scaling and root planing + application of 15% metronidazole gel (experimental). Treatment effects were assessed by longitudinal clinical parameters (gingival sulcus depth probing and clinical attachment level) and microbiological (analysis of subgingival microbiota by the technique of checkerboard DNA-DNA hybridization). The monitoring of concentrations of metronidazole in the gingival fluid collected from the patients receiving the gel was performed by high performance liquid chromatography. The results of the analysis of concentrations of metronidazole in periodontal pockets showed that the gel could only be detected at effective concentrations until one hour after application. Regarding microbiological analysis there was significant difference between groups in the three evaluated times (before, 7 and 30 days after treatment). The intragroup analysis showed a significant decrease in bacterial count only for the group treated with metronidazole gel after 7 days of treatment. Although difference was detected between both groups, none of them was able to eliminate or significantly reduce the periodontopathogenic species. With regard to clinical parameters, both therapies were effective in promoting reduction in depth probing and gain in attachment level, without significant benefit to the use of metronidazole gel.

When conspecific individuals are crossed, the ensuing hybridization creates a spectrum of phenotypes in the resulting offspring. Many of hybrid traits will be additive, similar to the parental phenotypes. In some cases however, transgressive phenotypes are formed, outside the range of that of the parental phenotypes. Transgressive phenotypes can either be restricted to the F1 generation or be heritable throughout the hybrid lineage. While the mechanism behind heritable transgressive phenotyped is yet to be determined, transgressive gene expression is thought to be the root cause of their formation. Epigenetics modifications, heritable variation separate to the DNA code, can alter gene expression, persist through generations, and vary between individuals and over time. This makes them ideal candidates to be involved in the formation of transgressive phenotypes. RNA silencing is an epigenetic mechanism of gene regulation relying on 20Q24nt single stranded small RNAs (sRNAs). Small RNAs, due to their ability to set up persistent epigenetic marks at a locus, have the potential to create heritable transgressive gene expression. For example, when genetic variation from one parental genome presents novel targets to the sRNAs of the other parental genome, new epigenetic marks such as DNA methylation or secondary sRNAs can be created at target sites. In order to understand the potential of small RNAs to influence hybrid phenotype, I designed crossing experiments with Chlamydomonas reinhardtii, choosing this unicellular alga due to the genetic tools available and the haploid nature of its vegetative cells. The specific aim of the experiment was to identify transgressively expressed sRNA populations. Crossing two geographically distinct strains of C. reinhardtii, and sequencing both the genomes and sRNAomes of parents and recombinants, I was able catalogue both genetic and epigenetic variation in the parental strains providing unique insight into the inheritance of small RNAs in this alga. In this thesis, I first compare the genomes of the parental strains, identifying polymorphisms and assessing genetic variation in RNA silencing pathway components. I then describe the sRNA profiles of the parental strains, identifying differentially expressed sRNA loci. I then describe my approach to identifying transgressively expressed sRNA loci in the hybrids. While many sRNA loci in the recombinants exhibit additive sRNA expression, I found multiple transgressively expressed sRNA loci. Using the available bioinformatics tools, I identified potential miRNAs and phased secondary sRNAs within the list of transgressively expressed loci. Target analysis of one of the transgressively expressed miRNAs linked it with the transgressive expression of certain phased loci, suggesting a potential for sRNAs to be able to set up heritable epigenetic marks in recombinant C. reinhardtii cells.

Made available in DSpace on 2016-04-25T20:20:39Z (GMT). No. of bitstreams: 1 Ivete Miranda Previtalli.pdf: 4422369 bytes, checksum: f531496965a7427fc5647397819f34ea (MD5) Previous issue date: 2012-05-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study proposes to reveal the candomblé angola circumscribed in the city of São Paulo, trying to comprehend the moment in which the search for affirmation of it´s identity couples up to the search of it´s African origins. This posture, manifested by some (pais de santo), promotes the dialogue as well as the confrontation among adepts of the Angola nation, pointing to the irreconcilable forces of tradition and translation . In this clash are revealed not only the hybridization process, in which sincretisms are recognised, but also what is no allowed to merge. This way, the oral stories collected from pais and mãe de santo, the observation of parties, the examination of old photographies and informal talks with the adepts, unveiled the questionings,anxieties and resistances that hold at the moment in the São Paulo Angola nation. The search for the imagined Africa can take many paths. But it may not produce africanisms, attest religious orthodoxies or separate nations into fixed frontiers. Besides this, to considerate that angola nation forms a religious continuum with the African Bantu traditional religions will end up dividing nations into pure and mixed , desconsidering constant transformations in ritual practices. In São Paulo, the Afro-Brazilian religious identity, has a cultural origin. The Africa that lives in the angola nation is one marked by diasporization and transformed in the New World. It is this imagined Africa produced by candomble´s narrative that may offer resources for the angola nation today. Even though, as only elements of culture with roots remain and flourish, we can not forget that the original elements were transformed by the process of cultural translation / Esse estudo propõe revelar o candomblé angola circunscrito na cidade de São Paulo, buscando compreender o momento em que a procura da afirmação de sua identidade atrela-se à busca de sua origem africana. Essa postura, manifestada por alguns pais de santo, suscita tanto o diálogo quanto a confrontação entre os adeptos da nação angola, apontando para as irreconciliáveis forças da tradição e da tradução . Nesse embate, revelam-se não somente os processos de hibridação, nos quais são reconhecidos os sincretismos, mas também o que não se deixa fundir. Assim, as histórias orais recolhidas dos pais e mães de santo, a observação das festas, o exame de fotografias antigas e as conversas informais com os adeptos, desvendaram os questionamentos, ansiedades e resistências que vigoram no momento na nação angola paulista. A procura da África imaginada pode tomar muitos caminhos. Mas ela não deve produzir africanismos, atestar ortodoxias religiosas ou separar as nações em fronteiras fixas. Além disso, considerar que a nação angola forma um continuum religioso com as religiões tradicionais da África banta acabará dividindo a nação entre os puros e os misturados , desconsiderando as constantes transformações nas práticas rituais. Em São Paulo, a identidade religiosa afro-brasileira, tem uma origem cultural. A África que vive na nação angola é aquela marcada pela diasporização e transformada no Novo Mundo. É essa África imaginada e produzida pela narrativa do candomblé que poderá fornecer recursos para a nação angola de hoje. Mesmo sabendo, que só permanecem ou florescem elementos de uma cultura que possuem raiz, não podemos esquecer que os elementos originais foram transformados pelo processo de tradução cultural

Made available in DSpace on 2016-04-26T14:53:35Z (GMT). No. of bitstreams: 1 Ivete Miranda Previtalli.pdf: 4422369 bytes, checksum: f531496965a7427fc5647397819f34ea (MD5) Previous issue date: 2012-05-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study proposes to reveal the candomblé angola circumscribed in the city of São Paulo, trying to comprehend the moment in which the search for affirmation of it´s identity couples up to the search of it´s African origins. This posture, manifested by some (pais de santo), promotes the dialogue as well as the confrontation among adepts of the Angola nation, pointing to the irreconcilable forces of tradition and translation . In this clash are revealed not only the hybridization process, in which sincretisms are recognised, but also what is no allowed to merge. This way, the oral stories collected from pais and mãe de santo, the observation of parties, the examination of old photographies and informal talks with the adepts, unveiled the questionings,anxieties and resistances that hold at the moment in the São Paulo Angola nation. The search for the imagined Africa can take many paths. But it may not produce africanisms, attest religious orthodoxies or separate nations into fixed frontiers. Besides this, to considerate that angola nation forms a religious continuum with the African Bantu traditional religions will end up dividing nations into pure and mixed , desconsidering constant transformations in ritual practices. In São Paulo, the Afro-Brazilian religious identity, has a cultural origin. The Africa that lives in the angola nation is one marked by diasporization and transformed in the New World. It is this imagined Africa produced by candomble´s narrative that may offer resources for the angola nation today. Even though, as only elements of culture with roots remain and flourish, we can not forget that the original elements were transformed by the process of cultural translation / Esse estudo propõe revelar o candomblé angola circunscrito na cidade de São Paulo, buscando compreender o momento em que a procura da afirmação de sua identidade atrela-se à busca de sua origem africana. Essa postura, manifestada por alguns pais de santo, suscita tanto o diálogo quanto a confrontação entre os adeptos da nação angola, apontando para as irreconciliáveis forças da tradição e da tradução . Nesse embate, revelam-se não somente os processos de hibridação, nos quais são reconhecidos os sincretismos, mas também o que não se deixa fundir. Assim, as histórias orais recolhidas dos pais e mães de santo, a observação das festas, o exame de fotografias antigas e as conversas informais com os adeptos, desvendaram os questionamentos, ansiedades e resistências que vigoram no momento na nação angola paulista. A procura da África imaginada pode tomar muitos caminhos. Mas ela não deve produzir africanismos, atestar ortodoxias religiosas ou separar as nações em fronteiras fixas. Além disso, considerar que a nação angola forma um continuum religioso com as religiões tradicionais da África banta acabará dividindo a nação entre os puros e os misturados , desconsiderando as constantes transformações nas práticas rituais. Em São Paulo, a identidade religiosa afro-brasileira, tem uma origem cultural. A África que vive na nação angola é aquela marcada pela diasporização e transformada no Novo Mundo. É essa África imaginada e produzida pela narrativa do candomblé que poderá fornecer recursos para a nação angola de hoje. Mesmo sabendo, que só permanecem ou florescem elementos de uma cultura que possuem raiz, não podemos esquecer que os elementos originais foram transformados pelo processo de tradução cultural

Made available in DSpace on 2016-04-26T18:18:48Z (GMT). No. of bitstreams: 1 Rogerio Luiz Covaleski.pdf: 3649593 bytes, checksum: e9fd1f5f39552e521cd87b73f6ac3ee4 (MD5) Previous issue date: 2010-07-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This research investigated how the process is occurring hybridization of advertising from the dialogue and the increasingly intense intersections between three actors in the contemporary media environment: the advertising market, the industry entertainment and interactive technologies. We had to describe and analyze paradigmatic ruptures of the current advertising model, the transition process creative and productive and trends for the advertising, in their new settings. In this thesis, we found the following hypotheses: 1) convergence media and media environment interactions generate new configurations advertising; 2) the inclusion in advertising language, elements such as entertainment and interactivity, leading to hybridization between those elements and functions of the original advertising - to inform and persuade; 3) the strategies of hybridization of language lead-in advertising to persuade, entertain and interact concurrently, making it a new media product: the interactive entertainment advertising. In research these hypotheses we have considered in various theoretical concepts and authors such as: cultural hybridity of Néstor García Canclini, which does not differ over what is cultured , massive or popular ; the route that generates the sense of complex narratives advertising, split between the manipulation , competence , performance and sanction , statement by José Luiz Fiorin; conceptual studies and genres of entertainment undertaken by researchers as Jeder Janotti Jr. and Itania Gomes; the types of interactivity surveyed by Pierre Lévy in relations with the messages communication devices; encouraging the sharing and dissemination of content from interactive media, researched by Karla Patriota. The object of research is related to advertising campaigns that involve branded content and/or branded entertainment together with the interactivity and opportunities for distribution via sharing. Five cases advertising make up the corpus of reading and analysis of this thesis: the commercial online "Carousel" for Philips winner of the Grand Prix at the International Festival Advertising Festival in 2009; the short film "Signs" a member of the Schweppes Short Film Festival; the advertising film "The Piano", for Nespresso, which comprises the hotsite What else? ; the micro "Gourmand em Casa," for Brastemp, consisting of episodes for TV and actions of interactivity on social networking website; the online shopping "The Hero", the Radiotjänst action customizable and encouraging dissemination. Parts of these campaigns fit into what we understand to be a new media product: the interactive entertainment advertising, as are hybrids, in the same effort communication, entertain, persuade, interact and are shared / Nesta pesquisa investigamos como está ocorrendo o processo de hibridização da publicidade a partir do diálogo e das intersecções cada vez mais intensas entre três atores do ambiente mediático contemporâneo: o mercado publicitário, a indústria do entretenimento e as tecnologias interativas. Tivemos como objetivos descrever e analisar as rupturas paradigmáticas do modelo publicitário vigente, as transições de processos criativos e produtivos e as tendências para a comunicação publicitária, em suas novas configurações. Nesta tese, verificamos as seguintes hipóteses: 1) a convergência midiática e as interações do ambiente mediático geram novas configurações publicitárias; 2) a incorporação, na linguagem publicitária, de elementos como entretenimento e interatividade, levam à hibridização entre aqueles elementos e as funções originais da publicidade informar e persuadir; 3) as estratégias de hibridização da linguagem publicitária levam-na a persuadir, entreter e interagir, concomitantemente, tornando-a um novo produto midiático: o entretenimento publicitário interativo. Na investigação destas hipóteses nos fundamentamos em diversos conceitos teóricos e autores como: o hibridismo cultural, de Néstor García Canclini, no qual não se distingue mais o que é culto, massivo ou popular; o percurso gerativo de sentido das narrativas complexas da publicidade, divididas entre a manipulação, a competência, a performance e a sanção, enunciado por José Luiz Fiorin; os estudos conceituais e de gêneros de entretenimento empreendidos por pesquisadores como Jeder Janotti Jr. e Itania Gomes; os tipos de interatividade pesquisados por Pierre Lévy nas relações das mensagens com os dispositivos de comunicação; o incentivo ao compartilhamento e à disseminação de conteúdos a partir de meios interativos, pesquisados por Karla Patriota. O objeto de pesquisa é relacionado às campanhas publicitárias que envolvem comunicação por conteúdo (branded content) e/ou entretenimento de marca (branded entertainment), aliadas à interatividade e às possibilidades de distribuição via compartilhamento. Cinco cases publicitários compõem o corpus de leitura e análise desta tese: o comercial online Carousel , para a Philips vencedor do Grand Prix no Festival Internacional de Publicidade de Cannes, em 2009; o curta-metragem Signs , integrante do Schweppes Short Film Festival; o filme publicitário Piano , da Nespresso, que compõe o hotsite What else?; a microssérie Gourmand em Casa , da Brastemp, composta por episódios para tevê e ações de interatividade em redes sociais na web; o comercial online The Hero , da Radiotjänst ação personalizável e com incentivo à disseminação. As peças destas campanhas se enquadram no que compreendemos ser um novo produto midiático: o entretenimento publicitário interativo, pois são híbridos que, em um mesmo esforço comunicacional, entretém, persuadem, interagem e são compartilháveis

Georges Lavaudant est un metteur en scène qui occupe une place importante dans l’histoire du théâtre français. Il a créé plus d’une centaine de spectacles pour le théâtre et l’opéra, en France et à l’étranger. Il a dirigé des théâtres, dès l’âge de 28 ans. Il est également auteur. Autodidacte, il s’est entouré d’une famille de travail – acteurs, décorateur, compositeur, chorégraphes – qui l’a accompagnée pendant quarante ans, des débuts en amateur aux scènes les plus prestigieuses. Il a mis en scène des classiques, des auteurs contemporains vivants, sa propre écriture. Son œuvre est dense et diverse. Il s’est, dès les débuts, positionné comme non-brechtien, préférant l’exploration du plateau à l’analyse dramaturgique a priori, préférant les formes non dramatiques à la fable didactique. Il considère la mise en scène comme un acte d’écriture. Lavaudant, passionné par la littérature et les écrivains, invente, avec les outils du théâtre, un imaginaire qu’il mêle à l’œuvre représentée. Cette thèse propose de découvrir le processus de création de ce metteur en scène, qui le mène à produire ce théâtre hybride avec lequel il transfigure le réel. / Georges Lavaudant is a director who occupies an important place in the history of French theater. He has created more than a hundred shows for theater and opera in France and abroad. He started directing theaters at the age of 28. He is also an author. Self-taught, he surrounded himself with a family of professionals - actors, a set designer, a composer, a choreographer - who accompanied him for forty years, from his debut as an amateur to the most prestigious stages. He has staged classic plays, living contemporary playwrights, and his own plays. His work is dense and diverse. From the beginning he was positioned as non-Brechtian, preferring the exploration of the stage to dramaturgical analysis a priori, and non-dramatic forms to the didactic fable. He considers staging as an act of writing. Lavaudant, who is passionate about literature and writers, uses the tools of the theater to invent an imaginary world that he combines with the work represented. This thesis investigates how this director’s creative process leads to this hybrid theater which transfigures reality.

Epigenetic regulatory mechanisms play important roles in the control of lineage-specific differentiation during development. However, mechanisms that regulate higher-order chromatin remodelling and transcription of keratinocyte-specific genes that are clustered in the genome into three distinct loci (Keratin type I/II loci and Epidermal Differentiation Complex (EDC)) during differentiation of the epidermis are poorly understood. By using 3D-Fluorescent In Situ Hybridization (FISH), we determined that in the epidermal keratinocytes, the KtyII and EDC loci are located closely to each other in the nuclear compartment enriched by the nuclear speckles. However, in KtyII locus knockout mice, EDC locus moved away from the KtyII locus flanking regions and nuclear speckles towards the nuclear periphery, which is associated with marked changes in gene expression described previously. Chromatin architectural protein Ctcf has previously been implicated in the control of long-range enhancer-promoter contacts and inter-chromosomal interactions. Ctcf is broadly expressed in the skin including epidermal keratinocytes and hair follicles. Conditional Keratin 14-driven Ctcf ablation in mice results in the increase of the epidermal thickness, proliferation, alterations of the epidermal barrier and the development of epidermal pro-inflammatory response. Epidermal barrier defects in Krt14CreER/Ctcf fl/fl mice are associated with marked changes in gene expression in the EDC and KtyII loci, which become topologically segregated in the nucleus upon Ctcf ablation. Therefore, these data suggest that Ctcf serves as critical determinant regulating higher-order chromatin organization in lineage-specific gene loci in epidermal keratinocytes, which is required for the proper control of gene expression, maintenance of the epidermal barrier and its function.

La recherche en contrôle a montré que le champ hospitalier français a subi de profonds changements, notamment à travers l’introduction de l’accountability. La conformité aux pressions institutionnelles pousse les organisations à s’orienter vers de nouveaux outils et principes pour renfoncer leur légitimité externe (Power, 2003). Cette même littérature a montré que l’accountability s’infiltre dans la sphère médicale et démontre des phénomènes d’hybridation de rôle chez les médecins hospitaliers, qui sont ainsi sensibilisés aux contraintes financières (Kurunmäki, 1999). La sphère administrative ne fait pas exception et subit également des pressions coercitives. L’objectif de notre thèse est de contribuer à cette littérature en analysant les effets et les rôles de l’accountability dans le secteur hospitalier public français sur la sphère médicale. Pour analyser ces effets, nous nous ciblons sur les dispositifs d’accountability tels que le reporting d’informations (audit financier, pratique budgétaire). Pour répondre à cette problématique, notre terrain s’est focalisé sur des entretiens semi-directifs et une approche ethnographique dans un CHU de grande taille. Premièrement, à travers une étude exploratoire qualitative menée auprès de 19 directeurs administratifs financiers (représentant 19 CHU/CH), nous montrons que l’accountability au travers de l’exemple de l’audit financier amène à des changements organisationnels et sert également à « rendre des comptes » aux parties prenantes externes (Agence Régionale de Santé, banques) mais aussi internes des organisations hospitalières. Nos résultats soulignent notamment que l’audit financier renforce la légitimité de la sphère administrative vis-à-vis de la sphère médicale. Deuxièmement, nous identifions à partir d’une approche ethnographique (observation non-participante et entretiens) menée dans un CHU des acteurs administratifs spécifiques, servant de vecteurs pour l’infiltration de l’accountability dans la sphère médicale : les Techniciens d’Information Médicale. Ces acteurs administratifs ont un rôle essentiel dans l’hybridation des médecins aux compétences gestionnaires, et les sensibilisent aux logiques financières. A travers le reporting de leur activité médicale, nos résultats mettent en lumière une atténuation du clivage traditionnel observé dans la littérature entre acteurs administratifs et corps médical. Nous montrons également que l’infiltration de l’accountability par d’autres agents administratifs spécialisés, tend à sensibiliser les médecins à la réduction de surplus, de slack présent dans la prise en charge des patients. Il s’avère que cette organisation hospitalière privilégie des dispositifs d’accompagnements afin de solliciter les médecins à intégrer de nouvelles responsabilités dans l’efficience de leur rôle. Notre étude montre que les médecins tentent désormais de trouver un équilibre afin d’articuler « logique patient » et logique gestionnaire. Pour cela, nos résultats soulignent une modification des protocoles de soins, du raisonnement ainsi que des valeurs des professionnels de santé qui concilient et promeuvent désormais le médico-économique. / How are accountability measures used by actors in the administrative field? Consequently, which are the associated effects on workers in the health sector?Research in control has shown that the French hospital has undergone profound changes, notably through the introduction of accountability. Compliance with institutional pressure drives organizations to move towards new tools and principles that reinforce their external legitimacy (Power, 2003). The same literature has shown that accountability infiltrates the medical sphere and that a new phenomenon occurs: doctors adopt hybridization role in hospital. This hybridization creates doctors who are aware of financial constraints (Kurunmäki, 1999). The administrative sphere is no exception and is also subject to coercive pressure.The aim of our thesis is to contribute to this literature by analyzing the intra-organizational effects and roles of accountability on the medical logic in the French public hospital sector. To analyze these effects, we focus on accountability mechanisms such as information reporting (i.e. financial audit, budget practice). To answer this problematic, we use semi-structured interviews into French hospitals and we adopt an ethnographic approach of one large university hospital.Firstly, from a series of exploratory interviews with 19 financial administrative directors (in 19 public hospitals), we show that accountability through financial audit leads to organizational changes and also use to “give account” not only to external stakeholders (Regional Health Agency, banks) but also internal hospital organizations. Our results show that the financial audit reinforces the legitimacy of the administrative sphere towards the medical sphere.Secondly, we identify with an ethnographic approach (non-participant observation and interviews) in a UHC of specific administrative actors called “Medical coders” (i.e. actors serving as vectors for the infiltration of accountability in the medical sphere). These administrative members have a key role in hybridization of health professionals to management skills, and sensitize them to financial logics.Through the reporting of their medical activity, our results highlight an attenuation of the traditional cleavage observed in the literature between administrative members and the medical profession.Moreover, we show that the infiltration of accountability by others specialized administrative agents tends to sensitize doctors to the reduction of surplus, and the slack in the care of patients. It turns out that, the hospital organization favors support systems to make physicians integrate efficiency in their health role.Our result show that physicians are now trying to find a balance in order to articulate medical logic and managerial logic. For this, our study highlights a modification of care protocols as well as the reasoning and values of health professionals who reconcile and promote medico-economic logic.This thesis argues that the introduction of accountability within the French public hospital sector has intra-organizational effects on both the administrative and medical spheres and emphasizes that on the basis of the reduction of the organizational slack, the conflicts between medical and managerial logic are lessened.