O objeto deste texto é o ruído em sua relação com a música. Minha ideia é investigar, nem sempre num percurso linear, o papel que ele desempenha na música, especialmente no período que vai do início do século XX aos dias de hoje. Essa investigação não está voltada para um levantamento técnico, nem historicista do tema. Minha preocupação está mais voltada a uma camada mais profunda em que transparecem algumas conexões entre o que chamamos de pensamento musical e a própria cultura que engendra esse pensamento. O ruído é um elemento essencial para o entendimento de certos aspectos da Modernidade, especialmente no que diz respeito à produção musical (e porque não dizer, à produção das artes em geral) no período. O que pretendo mostrar é que muitas das contradições e tensões que alimentam a música moderna podem ser entendidas como movimentos em direção à aceitação ou rejeição do ruído. A ideia central deste texto é a de que existe um processo dialético recorrente na relação entre música e ruído. Como algo incômodo, marginal, o ruído tende a ser evitado, pois sempre traz o risco de desagregação de um sistema. Minha intenção é investigar a passagem do ruído, enquanto elemento desorganizador, para som quando incorporado na música. Essa relação se direciona a uma compreensão da maneira como o ruído, no século XX, tornou-se um elemento desestabilizador, instaurando uma tensão dialética entre sua rejeição e sua aceitação enquanto elemento musical. / The object of this research is the noise and its relationship with music. My idea is to investigate, not always through linear path, the role it plays in music, especially in the period from early twentieth century to now. This investigation is not focused on a technical survey, nor on an historicist approach to the theme. My concern is more focused on a deeper layer in which one can find some connections between what we call \"musical\" and the very culture that generates it. Noise is an essential element for the understanding of certain aspects of modernity, especially in respect of music production (and why not to say, of art production in general) in that period. What I want to show is that many of the contradictions and tensions that feed modern music can be understood as a movement toward acceptance or rejection of noise. The central idea of this text is that there is a recurring dialectical process in the relationship between music and noise. As a disturbance, as a marginal element, noise tends to be avoided, because it always brings the risk of desegregating a system. My intention is to investigate the passage from noise as a disruptive element to music. This relationship is directed to an understanding of how noise has become a destabilizing element, establishing a dialectic tension between its rejection and acceptance as a musical element.

Esta tese apresenta o estudo dos casos de desligamentos de cinco professores da Escola de Comunicações e Artes da Universidade de São Paulo (ECA/USP) durante o regime militar (1969-1979): Jean Claude Bernardet, Thomaz Farkas, José Freitas Nobre, Jair Borin e José Marques de Melo. O objetivo desta pesquisa foi entender, por meio de documentos, depoimentos e pesquisa bibliográfica, a perseguição a intelectuais e cientistas na ditadura militar. O propósito foi realizar um estudo de caso para compreender como funcionava a lógica de investigações e controles dos expurgos nas universidades nesse contexto histórico brasileiro. Além disso, esta investigação tratou das diferentes estratégias de expurgos realizadas na USP, dentre elas as chamadas \"cassações brancas\", nas quais docentes não eram cassados formalmente, mas eram desligados dos quadros universitários sumariamente, sem justificativas. Para entender esses fenômenos, tratamos das trajetórias dos docentes desde o ingresso na ECA/USP, passando pelo desligamento, até o retorno na década de 1980 após a Lei da Anistia. Para tanto, utilizamos como base metodológica o método historiográfico, o paradigma indiciário e qualitativo de análise. Assim, dentro das discussões de que temos participado do Observatório de Comunicação, Censura e Liberdade de Expressão da ECA/USP, pudemos concluir que a intimidação e a autocensura foram preponderantes nas práticas de interdição dos dispositivos de vigilância nas universidades, gerando grandes consequências para a produção de conhecimento, principalmente no que se referiu à área de comunicação. / This thesis presents the study of the cases of dismissals of five professors of the School of Communications and Arts of the University of São Paulo (ECA/USP) during the military regime (1969-1979): Jean Claude Bernardet, Thomaz Farkas, José Freitas Nobre, Jair Borin and José Marques de Melo. The objective of this research was to understand, through documents, testimonies and bibliographic research, the persecution of intellectuals and scientists in the military dictatorship. The purpose was to carry out a case study to understand how the logic of investigations and controls of purges in universities in this Brazilian historical context worked. In addition, this investigation dealt with the different strategies of purges carried out at USP, among them the so-called \"white cassation\", in which teachers were not formally revoked, but were summarily dismissed from university staff without justification. In order to understand these phenomena, we have examined the trajectories of teachers since joining the ECA/USP, passing through the dismissal, until the return in the 1980s, after the Amnesty Law. For that, we use as methodological basis the historiographic method, the indiciary paradigm and qualitative analysis. Thus, within the discussions that we have participated in the Observatory of Communication, Censorship and Freedom of Expression of ECA/USP, we could conclude that intimidation and self-censorship were preponderant in the practices of interdiction of surveillance devices in universities, generating great consequences for the knowledge production, especially in the area of communications.

Desde seu descobrimento, o fenômeno de silenciamento gênico por RNA (RNAi) rapidamente se tornou uma técnica amplamente estudada e utilizada nos mais diversos aspectos da biologia molecular. Uma destas possibilidades é sua aplicação no campo da entomologia agrícola, mais especificamente para o controle de insetos-praga como uma alternativa de alta eficiência, especificidade e com impacto ambiental reduzido. Por meio da geração de plantas transgênicas expressando RNAi para genes essenciais de insetos-praga específicos, a ingestão destas moléculas de RNAi pelo inseto mediante herbivoria pode resultar no silenciamento do respectivo gene, resultando em fenótipos que podem variar entre perda de apetite, infertilidade ou até a morte. Neste contexto, o presente trabalho teve como objetivo provar a viabilidade de aplicação desta técnica para a interação Tomateiro x Tuta absoluta, cultura de grande expressão econômica e social no mercado nacional e internacional e que é amplamente atacada por esta praga, com prejuizos que podem alcançar a ordem dos 100% da produção. Por meio da clonagem de genes ortólogos essenciais descritos na literatura e de genes altamente expressos nos primeiros estádios larvais, após a caracterização transcriptômica em escala do inseto, foram realizados ensaios de alimentação contendo moléculas de dsRNAs que possuíam estes genes como alvo. Também, foi realizado a transformação genética de tomateiro cultivar \"Micro-Tom\" com dois destes genes (V-ATPase A e Arginina kinase) para a realização de ensaios de herbivoria. Com os resultados obtidos nestes experimentos, foram mostradas sólidas evidências da viabilidade da técnica de RNAi para o controle de Tuta absoluta, evidenciado pelo silenciamento gênico específico observado no inseto e consequentemente os efeitos nocivos deste silenciamento. / Since their discovery, the phenomenon of gene silencing by RNA ( RNAi ) has rapidly become a widely studied and used technique in the molecular biological field. One of these possible applications is in the entomology field, more specifically for the control of insect pests, as a high efficiency, specificity and with reduced environmental impact alternative. Through the generation of transgenic plants expressing dsRNA targeting essential insect genes, their ingestion by the insect and consequently the uptake of the silencing RNA, may result in specific gene silencing, resulting in a variety of phenotypes that can range from loss of appetite, infertility to death. In this context, this study aimed to prove the feasibility of this technique to control tomato leaf miner (Tuta absoluta) in tomatoes plant, a major crop worldwide and seriously attacked by this pest, with losses that can reach 100%. For the present work, orthologous genes from successfully cases of insect gene silence described in the literature, was selected together with highly expressed genes in the early larval stages of T. absoluta, chosen after the insect molecular characterization and used in feeding assays with dsRNAs molecules to targeted these genes. Also, genetic transformation of the \"Micro-Tom\" tomato cultivar with two of these genes (V-ATPase and Arginine kinase) was conducted for testing in an herbivore assay. With these two approaches was possible to get solid evidences of the feasibility of the RNAi technique to control this insect, evidenced by specific gene silencing observed and its consequent effect on pest phenotype.

A cana-de-açúcar (Saccharum spp.) é uma importante cultura na produção de alimentos e energia. Várias espécies de insetos podem causar sérios prejuízos econômicos à cultura da cana-de-açúcar. A broca da cana-de-açúcar (Diatraea saccharalis) é a praga de maior relevância por estar amplamente distribuída nas regiões canavieiras. O silenciamento gênico por RNA de interferência (RNAi) se tornou uma técnica amplamente estudada e utilizada nos mais diversos aspectos da biologia. Uma de suas aplicações é no controle de insetos-praga como uma alternativa de alta eficiência, especificidade e reduzido impacto ambiental. A ingestão de moléculas de RNA dupla fita (dsRNA) com identidade a regiões de genes essenciais de insetos-praga pode resultar no silenciamento destes genes, levando a fenótipos deficientes. Neste contexto, o presente trabalho teve como objetivo buscar genes alvos para o silenciamento com potencial para impedir o desenvolvimento normal da D. saccharalis e estabelecer uma forma de entrega do dsRNA eficiente para o teste de genes, visando assim validar o uso da técnica para a espécie. Por meio da clonagem de regiões de genes ortólogos já utilizados como alvo de silencimento em outras espécies de insetos (V-ATPase A, Receptor de Ecdisona e Arginina Kinase), e de genes com função específica identificadas após a caracterização do transcritoma de D. saccharalis (Juvenile Hormone Epoxide Hydrolase, Neverland e Quitina Sintase) foram conduzidos ensaios de RNAi. Foram realizados ensaios de dose resposta para o gene V-ATPaseem lagartas neonatas, onde a concentração selecionada por causar melhor redução na expressão do gene alvo foi de 2,5 µg µL-1. Esta concentração foi então utilizada em ensaios de alimentação para os outros genes. Os genes V-ATPase A, receptor de Ecdisona, Arginina Kinase, Juvenile Hormone Epoxide Hydrolase e Quitina Sintase apresentaram redução significativa no número de transcritos em larvas, demonstrando a viabilidade do uso de RNAi em D. saccharalisneonatas. O gene Neverland não demonstrou redução no acúmulo de transcritos nas condições trabalhadas. O gene GFP inicialmente utilizado como controle negativo apresentou variação na expressão de genes alvo, sendo desconsiderado como bom controle para D. saccharalis. O silenciamento dos genes alvo requer quantidades elevadas de dsRNA, superiores aos obtidos por transcrição in vitro, o que limita a viabilidade de ensaios com maiores replicatas e para determinar efeitos biológico. Alternativas de produção de dsRNA devem ser avaliadas para viabilizar a seleção de genes alvo efetivos / Sugarcane (Saccharum spp.) is an important crop for the production of food and bioenergy. Many insect species can cause economic losses in sugarcane. The sugarcane borer (Diatraea saccharalis) is the most important sugarcane pest, because it occurs in all production regions. Gene silencing by RNA interference (RNAi) rapidly became a widely investigated approach, adopted in various aspects of biology. One of the potential applications of RNAi is agricultural pest control, as an alternative with high efficiency, specificity and reduced environmental impact. The ingestion of double-stranded RNA (dsRNA) molecules with identity to regions of essential genes of the insect-pest can result in the target gene knock-down and, consequently, to deficient phenotypes. In the present work, target genes with the potential to affect the normal development of D. saccharaliswere searched, together with an efficient dsRNA delivery approach to test the target-genes to validate the use of the RNAi in D. saccharalis. Based on degenerated primers, expressed orthologous genes previously tested in other insect species (V-ATPase A, Ecdisone Receptor, and Arginine Kinase) were cloned,whilegenes with specific function (Juvenile Hormone Epoxide Hydrolase, Neverland, and Chitin Synthase) were identified from an in-house assembled transcriptome of D. saccharalis and cloned. A dose-response assay was conducted using the V-ATPase gene region delivered by droplets to neonate larvae, and the 2.5 µg µL-1dsRNA concentration was selected for further tests. This concentration was then used to deliver the other genes. The dsRNA version from the genes V-ATPase A, Ecdisone Receptor, Arginine Kinase, Juvenile Hormone Epoxide Hydrolase and Chitin Synthaseexhibited a significant reduction in the accumulation of transcripts, indicating the viability of RNAi to D. saccharalis in 1st instar larvae. The Neverland gene was not silenced by RNAi in the used conditions. The dsRNA of the Green Fluorescent Protein gene, used as negative control appeared to affectother gene targets. Target gene silencing require large amounts of dsRNA, more than what is achievable by in vitro transcription, which limits the viability to conduct large assays with more replicates and to determine biological effects. Alternatives to produce dsRNA need to be evaluated to enable the selection of effective target genes

La présence de domaines chromosomiques heterochromatiniens associé à des effets de position est une propriété communes à de nombreux génomes eukaryotes. L'intensité et l'étendue de la variégation liée aux effets de position sont généralement sensibles à la dose des protéines effectrices de l'hétérochromatine. Les propriétés d'auto-propagation des complexes d'hétérochromatine a un cout, qui est la nécessité d'établir des mécanismes stoppant la propagation de la répression transcriptionelle. Cette thèse explore la dose-dépendance de l'effet de position télomérique en étudiant le complexe SIR de la levure du boulanger. La caractérisation du groupement des télomères en foyers, de la localisation de Sir3 et de la transcription dans des souches sur-exprimant Sir3 a permis d'établir l'étendue maximale des domaines silencieux présent aux subtelomeres. L'étude de jeux de données publiés a révélé que ces domaines terminent généralement au niveau de zones correspondant où les propriétés de la chromatine montrent une transition importante. Ces transitions chromatiniennes sont requises pour survivre en présence d'un excès de protéines Sir3 puisque nous avons démontré que les mutants dot1 ne survivent pas un tel excès. En outre nous avons conduit un crible génétique qui a révélé de nombreux gènes requis pour la survie en présence d'une surdose de Sir3. Ce travail caractérise la réponse du génome à une surdose d'hétérochromatine et a permis de révéler des domaines subtélomeriques associés à des propriétés chromatiniennes particulières. En conséquence nous démontrons comment l'effet de position télomerique est efficacement restreint au subtelomere chez la levure. / A shared property of several eukaryotic genomes is the presence of heterochromatic chromosomal domains experiencing transcriptional variegation. The intensity and the extent of position effect variegation are sensitive to the dosage of silencing effectors in many systems. The self-propagating properties of heterochromatin machineries come with a cost, which is the requirement for mechanisms preventing ectopic spreading of silencing. This thesis explores the dose-dependency of telomere position effect, using the budding yeast SIR system as a model for chromatin based heterochromatic silencing. To assess the dose-dependency of telomere position effect in budding yeast, we systematically characterized the impact of Sir3 overexpression by quantifying the clustering of telomeres, the genome wide binding of Sir3 and its impact on coding and non coding transcription. Analysis of published data sets enabled to uncover candidates potentially responsible for the limitation of subtelomeric silent domains. Our study reveals that extension of silent domains can reach saturation, associated with the anti-silencing properties of histone marks deposited by the conserved enzyme Dot1. In addition we discovered genes required for viability upon SIR3 overexpression by conducting a genetic screen. Our work describes the dynamics of the dose dependency of heterochromatin propagation in budding yeast. It uncovers previously uncharacterized discrete chromosomal domains associated with specific chromatin features and demonstrates how telomere position effect is efficiently restricted to subtelomeres by the preexisting chromatin landscape.

CYCLOIDEA (CYC) and DICHOTOMA (DICH) of the CYC2 clade of the TCP gene family have been shown to play a significant role in regulating the identity of the dorsal petals and abortion of the single dorsal stamen in Antirrhinum majus. It is believed that CYC2-like genes are responsible for the convergent evolution of floral zygomorphy, but their role in the development of actinomorphic flowers is still unknown. In Solanaceae, previous analysis has identified two paralogs of CYC2-like genes, CYC2A and CYC2B, resulting from a gene duplication that predates the origin the family. Virus-induced gene silencing (VIGS) is a technique to study the gene function by silencing specific target genes of interest, which is shown to be useful in diverse plant species. Here, we report on the role of CYC2-like genes during floral development in Solanaceae based on the results of VIGS using tobacco rattle virus (TRV)-based vector in Solanum lycopersicum having completely actinomorphic flowers and Nicotiana obtusifolia having slightly zygomorphic flowers. Our VIGS experiments in So. lycopersicum show that downregulation of both CYC2A and CYC2B leads to misshaped petals, the unequal growth of the petals, and most frequently increased number of petals, stamens and sepals, while the carpel and ovule morphology remain the same as the wild type. On the contrary, downregulation of CYC2A and CYC2B in N. obtusifolia results in reduced number of flower organs in sepals, stamens, and petals, however carpels remained the same. For both solanaceous species, silencing CYC2A and CYC2B changes the property of cytoplasm and retards the rate of pollen germination. Our findings suggest that the CYC2-like genes are likely involved in the floral development, mainly regulating the number of floral organs and pollen development in Solanaceae.

Oncolytic viruses are novel candidates for cancer therapy and their efficacy relies on their capacity to overcome the host’s anti-viral barriers. In mammalian cells, the anti-viral response involves a protein-signaling cascade known as the interferon pathway, which alerts the immune system and limits the propagation of infection. Given that most cancer cells have defects in this pathway, they are susceptible to viral infection and responsive to oncolytic virotherapy. For reasons that remain unknown, many cancers are still refractory to oncolytic viruses, which suggests the existence of additional antiviral mechanisms. In this study, we investigate the potential involvement of an alternative antiviral pathway in cancer cells. Given that insects and plants rely on the RNA silencing pathway for their anti-viral protection, we investigated the presence of a similar mechanism in cancer cells. We found viral genome-derived small RNAs in various cancer cell lines upon infection, which is indicative of an RNA-mediated antiviral response. Also, various viruses encode suppressors of the RNA interference pathway. To determine if an oncolytic virus could benefit from such a factor, we engineered an oncolytic virus variant to encode the Nodamura virus B2 protein, a known inhibitor of RNA silencing-mediated immune responses. Using this virus, we observed enhanced cytotoxicity in 33 out of the 38 human cancer cell lines tested. Furthermore, our results show inhibition of viral genome cleavage and altered microRNA processing by our B2-expressing oncolytic virus. Taken together, our data suggests the blockade of RNA silencing antiviral pathways and/or antiviral microRNA processing improves the efficacy of our B2-encoding virus in a cell-line specific manner. Overall, our results establish the improved potential of our novel virus therapy and demonstrate for the first time the involvement of RNA pathways in the antiviral defense of cancer cells.

La maladie de Huntington (MH) est causée par une expansion de répétitions CAG sur l’exon 1 du gène huntingtine (htt), codant pour une protéine mutée. Il a été montré que la diminution d’expression de cette protéine est une piste thérapeutique très prometteuse. Dans ce projet, nous avons étudié et comparé trois approches dites «antisens» : une stratégie allèle non-spécifique, visant à diminuer de manière générale l’expression de htt ; une stratégie allèle spécifique ciblant les répétitions CAG afin d’impacter préférentiellement l’allèle muté ; et enfin une stratégie de saut d’exon permettant d’enlever des sites de clivage à l’origine d’une forme raccourcie et toxique de la protéine htt. Nous avons évalué ces approches grâce à deux outils différents : les tricyclo-DNA (TcDNA), qui sont une nouvelle classe d’oligonucléotides antisens (AON) plus performante que les chimies précédentes, et le système U7snRNA vectorisé, permettant d’induire une expression stable des séquences antisens. Dans un premier temps, ces différentes molécules ont été évaluées in vitro dans des lignées de fibroblastes de patients en quantifiant le niveau d’ARNm et de protéines htt par RTqPCR et Western blot respectivement. Par la suite, les séquences les plus efficaces in vitro ont été sélectionnées et les AON et AAV-U7snRNA correspondants ont été injectés en intracérébroventriculaire (ICV) dans un modèle murin de la MH (souris YAC128). Les résultats les plus encourageants ont été obtenus avec le TcDNA-NS (pour allèle Non Spécifique), permettant une diminution significative de l’expression de htt dans le cortex, l’hippocampe et le striatum 2 et 6 semaines après une injection ICV. Ces résultats prometteurs suggèrent le potentiel des TcDNA comme nouvel outil thérapeutique pour la MH. / Huntington’s disease (HD) is caused by a CAG repeat expansion in the exon 1 of huntingtin gene (htt), encoding for a mutant protein. It has been shown that the silencing/down regulation of huntingtin protein is a promising therapeutic lead. In this project, I have explored and compared three strategies using the antisense approach: a non-allele specific strategy, aiming to silence the global expression of htt; an allele specific strategy targeting CAG repeats to silence preferentially the mutant allele; and an exon-skipping strategy in order to remove cleavage sites which originally cause a shorter and toxic form of the htt protein. These strategies have been evaluated using two different tools: tricyclo-DNA (TcDNA), a new class of antisense oligonucleotides (AON) more efficient than the previous chemistries, and a vectorized approach using U7snRNA system allowing a stable expression of antisense sequences. Firstly, these different molecules have been assessed in vitro in HD fibroblasts quantifying mRNA and htt protein levels with RTqPCR and Western blot respectively. Subsequently, the most efficient sequences have been selected and intracerebroventricular (ICV) injections have been performed with corresponding AON and AAV-U7snRNA in a HD mouse model (YAC128). The most encouraging results have been obtained with the TcDNA-NS (for Non Specific allele), allowing a significant decrease of htt expression in cortex, hippocampus and striatum 2 and 6 weeks after ICV injection. These promising results suggest the potential of TcDNA as a new therapeutic tool for HD.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22727号 / 医博第4645号 / 新制||医||1045(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 濵﨑 洋子, 教授 羽賀 博典, 教授 伊藤 貴浩 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

In all common hydrostatic pumps, compressibility affects the commutation phases of the displacing chambers, as they switch their connection from/to the inlet to/from the outlet port, leading to pressure peaks, localized cavitation, additional port flow fluctuations and volumetric efficiency reduction. In common pumps, these effects are reduced by proper grooves that realizes gradual port area variation in proximity of these transition regions. This paper presents a method to automatically find the optimal designs of these grooves, taking as reference the case of external gear pumps. The proposed procedure does not assume a specific geometric morphology for the grooves, and it determines the best feasible designs through a multi-objective optimization procedure. A commercial gear pump is used to experimentally demonstrate the potentials of the proposed method, for a particular case aimed at reducing delivery flow oscillations.