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<p>The Gram-positive, soil dwelling bacteria of the genus <em>Streptomyces</em> produce greater than 50% of the clinically relevant antibiotics in use today. Thanks to the falling price of DNA sequencing, <em>Streptomyces</em> genomes are revealing that they encode more secondary metabolites (potential antibiotics) than they produce under standard laboratory conditions. By heterologously overexpressing the known pleiotropic regulators of antibiotic expression from <em>Streptomyces coelicolor</em> in several other <em>Streptomyces</em> species it has been shown that the secondary metabolite profile of these species can be influenced. While present-day methods of introducing genes (conjugation) and screening for antibiotics work well on a small scale, the low throughput nature of these protocols stand as a barrier to testing this hypothesis on a larger scale. The focus of the research presented here was to develop high throughput (HTP) methods of engineering and screening <em>Streptomyces</em>. With these two technologies in place, an attempt was to made to introduce three plasmids (pSET152-<em>ermE*</em>p-null, pSET152-<em>ermE</em>*p-<em>atrA</em> and pSET152-<em>ermE</em>*p-<em>lsr2_NTD</em>) into 120 wild-isolate <em>Streptomyces</em> species from the Wright Actinomycete Collection. Exconjugants were successfully obtained for all three plasmids in 48 species of <em>Streptomyces</em> and were screened for increased antimicrobial activity using a HTP, <em>lux</em>-based bioassay. Numerous strains showed increased antimicrobial activity but WAC00206, WAC00230 and WAC00263 with pSET152-<em>ermE</em>*p-<em>lsr2_NTD</em>showed the most promising improvement in antimicrobial activity. These hits have been designated as high priority for future investigation. These results suggest that HTP conjugation and the <em>lux</em>-based bioassay are powerful methods for introducing plasmids into and screening engineered streptomycetes.</p> / Master of Science (MSc)

Orientador: Danielle Biscaro Pedrolli / Resumo: Os métodos de indução da expressão gênica disponíveis para linhagens bacterianas envolvem a adição de compostos indutores ao meio de cultura (por exemplo, isopropil β-D-1-tiogalactopiranosideo, xilose e arabinose), o que é indesejável para linhagens industriais, pois encarece o processo produtivo. Já a utilização da expressão constitutiva, alternativa à indução, pode ocasionar stress metabólico durante a fase lag de crescimento quando são utilizados promotores fortes. O objetivo do trabalho foi construir e padronizar um novo modelo de indução da expressão gênica para linhagens bacterianas industriais. O novo modelo de autoindução baseado no sistema de quorum-sensing bacteriano, permitindo que a célula se automonitore e induza a expressão gênica durante a fase exponencial de crescimento, eliminando assim não só a necessidade de adição de composto indutor como a necessidade de monitoramento da densidade celular pré-indução. Realizou-se amplificação e clonagem dos genes luxR e luxI, com e sem caudas de histidina, e suas respectivas sequências regulatórias de Aliivibrio fischeri, em plasmídeo contendo os genes responsáveis pela bioluminescência ou fluorescência com códons otimizados para Bacillus subtilis. Em seguida, foi realizada transformação e a integração do plasmídeo no cromossomo de B. subtilis. A funcionalidade do sistema foi avaliada em diferentes etapas de crescimento microbiano com o auxílio de um leitor de microplacas durante intervalos regulares. O sistema de autoind... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Current methods available for the autoinduction of gene expression in genetically engineered bacterial strains require addition of inducing compounds to the culture medium (e.g. Isopropyl β-D-1-thiogalactopyranoside, xylose, and arabinose), which is undesirable for industrial strains due to additional costs to the production process. Alternatively, constitutive gene expression is employed. However, the later can possibly cause metabolic stress during the lag growth phase if strong promoters are employed. The objective of this work was to construct and to standardize a new model for induction of gene expression in industrial bacterial strains. The new model is based on an autoinduction process triggered by the bacterial quorum-sensing system. It allows the cell to monitor itself and induce its own gene expression during the exponential growth phase, thereby eliminating both the need for an external inducing compound and the need for monitoring pre-inducing cell density. Bacterial cultures were grown in rich media, supplemented or not with antibiotics. Amplification and cloning of luxR and luxI genes, with and without histidine tags, and their respective regulatory sequences of Aliivibrio fischeri, were performed on a plasmid containing the genes responsible for bioluminescence or fluorescence with codons optimized for Bacillus subtilis. Next, transformation and integration of the plasmid into the B. subtilis chromosome were performed. The functionality of the system was evalua... (Complete abstract click electronic access below) / Doutor

A Antologia do conto russo, publicada pela Editôra Lux Ltda. no início dos anos 1960 e organizada por Otto Maria Carpeaux, é a primeira tradução de contos russos realizada diretamente da língua de origem. Este estudo resgata as informações e a situação em que essa coletânea de textos foi criada, numa tentativa de remontar seu contexto histórico, momento literário e mercado editorial, buscando hipóteses que auxiliem no entendimento dos aparatos que a constituem, num esforço de aproximação do escasso material de divulgação localizado sobre obra, editora e organizador pesquisado nos periódicos da época e constante dos anexos , numa reflexão sobre os laços de interesse existentes entre o Brasil e a União Soviética. / The Anthology of Russian Short Stories, edited by Otto Maria Carpeaux and published in the early 1960s by Editôra Lux Ltda, marks the first time Russian short stories were published from direct translation of the original language to Portuguese. This study retrieved information about the Brazilian historical and literary context as well as its publishing market during the time the work was created in an effort to promote a better understanding of how it was influenced by them. The presented information about the work, publisher and editor )included in the attachments( was searched in the sparse number of publishing advertisements in the newspapers and magazines that circulated during that time. It allows a better understanding of the works content and promotes a deeper reflection on the common interests and relationship between Brazil and the USSR.

Bonaventure et la lumière : la question semble au premier abord et pour la plupart des médiévistes résolue. Pourtant, la genèse de sa définition de la lumière (II Sent., d. XIII) – notamment la manière dont les idées de Robert Grosseteste lui parviennent – reste encore à déterminer. L’idée d’un usage métaphorique et analogique de la lumière et de sa dimension épistémique permet de mieux évaluer la théorie de la connaissance comme lumière, c’est-à-dire son amplitude réelle souvent réduite à l’illumination divine de l’homme tant intellectuelle que morale. Cette connaissance de la lumière permettant de considérer la connaissance comme lumière par le truchement de la métaphore et de l’analogie donc toute une épistémologie par la lumière se vérifie dans la cosmologie et dans la théorie de la beauté du frère mineur où la lumière joue bien un rôle principiel et paradigmatique. Enfin, que toutes ces implications philosophiques et théologiques de la lumière soient récapitulées dans le Christ compris selon les Écritures, comme splendor, sol iustitiae, lux mundi, compréhension nettement inspirée de la lecture du quatrième évangile et du Livre de la Sagesse par Bonaventure, bachelier biblique, demande encore à être élucidé. C’est en cherchant tant du côté des études à la Faculté des arts de Paris de 1235-1243 que du côté des écrits de Bonaventure, bachelier biblique puis sententiaire, que la question de la lumière dans son œuvre peut être interprétée. Nous proposons donc dans cet ouvrage d’ouvrir quelques pistes de compréhension de la via lucis bonaventurienne. / Bonaventure and light: the issue seems at first and for most medievalists resolved.Yet the establishment of the genesis of his definition of light (II Sent., d. XIII) — and in particular the way in which Robert Grosseteste's ideas reach him — is still to be determined. The idea of a metaphorical and analogical use of light and its epistemic dimension makes it possible to evaluate better the theory of knowledge as light, that is, its actual amplitude, often reduced to the divine enlightenment of Man, both intellectual and moral. This knowledge of light, which makes it possible to consider knowledge as light through metaphor and analogy, and thus a whole epistemology by light, is verified in the cosmology and in the theory of beauty of the Friar Minor in which light plays indeed a principle-like and paradigmatic role. Finally, that all these philosophical and theological implications of light are recapitulated in Christ understood according to the Scriptures as splendor, sol iustitiae, lux mundi, an understanding clearly inspired by the reading of the fourth Gospel and the Book of Wisdom by Bonaventure, bachelor of the Bible, still needs to be elucidated. It is by seeking as much on the side of studies at the Faculty of Arts of Paris from 1235 to 1243 that on the writings of Bonaventure, as baccalaureus biblicus and then baccalaureus sententiarus, that the question of light in his work can be interpreted. We therefore propose in this book to open some avenues of understanding of the bonaventurian via lucis.

A Antologia do conto russo, publicada pela Editôra Lux Ltda. no início dos anos 1960 e organizada por Otto Maria Carpeaux, é a primeira tradução de contos russos realizada diretamente da língua de origem. Este estudo resgata as informações e a situação em que essa coletânea de textos foi criada, numa tentativa de remontar seu contexto histórico, momento literário e mercado editorial, buscando hipóteses que auxiliem no entendimento dos aparatos que a constituem, num esforço de aproximação do escasso material de divulgação localizado sobre obra, editora e organizador pesquisado nos periódicos da época e constante dos anexos , numa reflexão sobre os laços de interesse existentes entre o Brasil e a União Soviética. / The Anthology of Russian Short Stories, edited by Otto Maria Carpeaux and published in the early 1960s by Editôra Lux Ltda, marks the first time Russian short stories were published from direct translation of the original language to Portuguese. This study retrieved information about the Brazilian historical and literary context as well as its publishing market during the time the work was created in an effort to promote a better understanding of how it was influenced by them. The presented information about the work, publisher and editor )included in the attachments( was searched in the sparse number of publishing advertisements in the newspapers and magazines that circulated during that time. It allows a better understanding of the works content and promotes a deeper reflection on the common interests and relationship between Brazil and the USSR.

A sociedade contemporânea é marcada pelo consumo, pelas estratégias mercadológicas que, muitas vezes, são efêmeras e só tentam satisfazer uma pequena parte da nossa constante procura pela completude. A publicidade está inserida nesse contexto ao expressar os valores vigentes na sociedade e também em mediar as possibilidades de inserção dos consumidores nos mais diversos nichos e estratos sociais. As marcas procuram expressar valores, identidades e imagens dos produtos/serviços por meio de ações e manifestações. Tais ações e manifestações podem vir de diversas maneiras, ou seja, em anúncios de mídia impressa, ou na forma de audiovisual; ações de merchandising como as vitrines, as gôndolas, dentre outras e que têm como intuito a geração de vínculos positivos, de associações prazerosas e levar o potencial consumidor ao ato e usufruto do bem ou serviço. O presente trabalho resulta de uma pesquisa entre as marcas de luxo em comparação das marcas mainstream, um aprofundamento intersígnico procurando identificar os efeitos de sentido de cada, suas semelhanças e distinções. O trabalho tem como objetivo compreender a circulação intersígnica das marcas de luxo e das marcas mainstream no contexto do varejo de moda. Os resultados que aqui compartilharemos é proveniente de uma pesquisa que conta com o intercruzamento dos campos da comunicação com o a antropologia visual. A comunicação, mas precisamente, a publicitária, entrará como sua retórica de consumo por meio dos signos que se relacionam para uma produção de sentido eficaz. Já a antropologia visual ficará por conta da pesquisa fotoetnográfica que nos auxiliará no mapeamento e narrativa do consumo de luxo e do mainstream. Como método de análise nos apoiaremos na Semiótica desenvolvida por Charles Sanders Peirce. O registro fotoetnográfico foi realizado nas cidades de: São Paulo, Fortaleza, Brasília e Florianópolis e buscamos fotografar as vitrines das marcas e luxo e das marcas mainstream que nos possibilitem um estudo sobre a circulação intersígnica entre as mais variadas vitrines. Este trabalho não terá como pretensão a análise de recepção por parte dos consumidores, ficaremos restritos só aos possíveis efeitos de sentido que as marcas se utilizam para ampliar seu escopo imagético e identitário. / ephemeral and only try to satisfy a small part of our constant search for completeness. Publicity is inserted in this context when expressing the values in force in society and also in mediating the possibilities of insertion of the consumers in the most diverse niches and social strata. Trademarks seek to express values, identities and images of products / services through actions and manifestations. Such actions and manifestations may come in a variety of ways, that is, in print media advertisements, or in the form of audiovisuals; Merchandising actions such as shop windows, gondolas, among others and whose purpose is to generate positive bonds, pleasant associations and lead the potential consumer to the act and usufruct of the good or service. The present work results from a research between the luxury brands in comparison of the mainstream brands, a deepening intersígnico looking for to identify the effects of sense of each, its similarities and distinctions. The work aims to understand the intersection of luxury brands and mainstream brands in the context of fashion retail. The results we share here come from a research that relies on the cross-linking of the fields of communication with visual anthropology. Communication, but precisely the advertising, will enter as its rhetoric of consumption through the signs that relate to a production of effective meaning. The visual anthropology will be based on the photoetnographic research that will help us in the mapping and narrative of the consumption of luxury and the mainstream. As a method of analysis we will rely on the Semiotics developed by Charles Sanders Peirce. Photoetnographic registration was carried out in the cities of: São Paulo, Fortaleza, Brasília and Florianópolis, and we sought to photograph the showcases of the brands and luxury and of the mainstream brands that allow us to study the intersection circulation among the most varied shop windows. This work will not have as a pretension the analysis of reception by consumers, we will be restricted only to the possible effects of meaning that the brands are used to expand their image and identity scope.

Christian Louboutin é um designer da moda, que trata da estética sob a ótica do processo criativo, um produtor de artigos de luxo, que atua em um nicho de mercado com acessórios da moda de luxo, para homens e mulheres com alto poder aquisitivo. Faremos um estudo de caso único, delimitado ao assunto da estética e o design nos sapatos femininos da marca Christian Louboutin, a fim de buscarmos respostas através de alguns questionamentos para as seguintes questões de pesquisa: 1) Como a estética e o design nos sapatos da marca Christian Louboutin desempenham um papel importante no despertar da imagem, de poder, da elegância, da sensualidade e do fetiche nas clientes Louboutin? 2) Como os elementos resultantes dos sentimentos de prazeres e das experiências sensoriais subliminares estimuladas por atributos estéticos, dão origem a diversas sensações e emoções que servem de incentivo no processo de compra? O objetivo deste estudo será o de encontrar os principais atributos estéticos que expressam determinado nível de beleza nos sapatos Christian Louboutin, aqueles que resultam em experiências emocionais sensoriais subliminares que influem nos gostos e nos gastos das clientes, quando compram os sapatos de luxo da marca. A metodologia deste trabalho consistirá de um estudo de caso único que utilizará os métodos de coleta de dados: observação e entrevista. As entrevistas serão realizadas com seis clientes no ponto de venda, associada à determinada fundamentação teórica. Os resultados obtidos demonstraram que não ocorreram divergências significativas nas respostas dadas pelas entrevistadas se contrapomos aos objetivos deste trabalho. Os resultados das entrevistas se alinham ao conceito de estética e do design dado pela fundamentação teórica, e por serem considerados os elementos mais admirados pelas clientes nos sapatos de luxo Christian Louboutin. A conclusão deste estudo de caso é que os elementos estéticos encantam e fascinam as clientes, despertam sensações de poder, paixões e fetiche, além de dar a sensação de ficarem mais lindas, sensuais e atraentes. A estética age como um elemento imprescindível à inovação e a diferenciação nos sapatos, garante a autenticidade da marca, servindo de conexão como o imaginário de suas clientes. / Christian Louboutin is a fashion designer who deals with the aesthetics from the perspective of the creative process, a producer of luxury goods, which operates in a niche market with luxury fashion accessories for men and women with high purchasing power. We\'ll make a single case study, defined the subject of aesthetics and design in women\'s shoes brand Christian Louboutin, to seek answers through some questions to the following research questions: 1) How the aesthetics and design in shoes brand Christian Louboutin play an important role in the image of awakening, power, elegance, sensuality and fetish in Louboutin customers? 2) How the resulting elements of the feelings of pleasure and subthreshold sensory experiences stimulated by aesthetic attributes, give rise to different sensations and emotions that serve to encourage the purchase process? The aim of this study is to find the main aesthetic attributes that express certain level of beauty in Christian Louboutin shoes, those that result in subliminal sensory emotional experiences that influence the tastes and spending of customers when they buy luxury brand shoes. The methodology of this work will consist of a single case study that will use the data collection methods: observation and interview. The interviews will be conducted with six customers at point of sale, associated with a particular theoretical framework. The results showed that no significant differences in the answers given by the respondents to counterpose the objectives of this work. The results of the interviews are aligned to the concept of aesthetics and design given by the theoretical basis, and consider the most admired elements by customers in luxury shoes Christian Louboutin. The conclusion of this case study is that the aesthetic elements delight and fascinate customers, arouse feelings of power, passion and fetish, and give the feeling of being the most beautiful, sexy and attractive. The aesthetic acts as an essential element for innovation and differentiation in shoes, guarantees the authenticity of the brand, serving as a connection to the imagination of its customers. Google Tradutor para empresas:Google Toolkit de tradução para appsTradutor de sitesGlobal Market Finder

Since its characterization, the definitive shortcoming of the bacterial luciferase (lux) bioluminescent reporter system has been its inability to express at a functional level in the eukaryotic cellular background. While recent developments have allowed for lux function in the lower eukaryote Saccharomyces cerevisiae, they have not provided for autonomous function in higher eukaryotes capable of serving as human biomedical proxies. Here it is reported for the first time that, through a process of poly-bicistronic expression of human codon-optimized lux genes, it is possible to autonomously produce a bioluminescent signal directly from mammalian cells. The low background of the bioluminescent signal, along with its characteristic lack of substrate amendment required for bioluminescent production, makes a mammalian-based lux reporter system ideal for real-time monitoring of cell culture or murine model systems. The delectability of a lux-based system provides for a functionally equivalent process to monitoring firefly luciferase-expressing cells under cell culture or subcutaneous imaging conditions without the well-documented uncertainties stemming from additional substrate introduction. However, the relatively blue-shifted emission wavelength of the lux reporter system, along with its low quantum yield, has been shown to reduce its effectiveness for use during deep tissue imaging of animal subjects. Despite these disadvantages, it has been demonstrated that a human cell line expressing the human codon-optimized lux genes can function as a biosensor for determination of human bioavailability of toxic compounds and that, by regulating the production of the luxC and luxE genes, the lux system can be employed as the first mammalian, real-time, fully autonomous bioreporter. These cell lines provide unique and efficient models for the detection and monitoring of human-relevant compounds of interest. The limiting reagent for bioluminescent production in the mammalian cellular background has been determined to be the cytosolic availability of the FMNH2 co-substrate and, in light of this evidence, directions for future optimization have been characterized and evaluated in respect to their ability to increase bioluminescent yield under these conditions.

A sociedade contemporânea é marcada pelo consumo, pelas estratégias mercadológicas que, muitas vezes, são efêmeras e só tentam satisfazer uma pequena parte da nossa constante procura pela completude. A publicidade está inserida nesse contexto ao expressar os valores vigentes na sociedade e também em mediar as possibilidades de inserção dos consumidores nos mais diversos nichos e estratos sociais. As marcas procuram expressar valores, identidades e imagens dos produtos/serviços por meio de ações e manifestações. Tais ações e manifestações podem vir de diversas maneiras, ou seja, em anúncios de mídia impressa, ou na forma de audiovisual; ações de merchandising como as vitrines, as gôndolas, dentre outras e que têm como intuito a geração de vínculos positivos, de associações prazerosas e levar o potencial consumidor ao ato e usufruto do bem ou serviço. O presente trabalho resulta de uma pesquisa entre as marcas de luxo em comparação das marcas mainstream, um aprofundamento intersígnico procurando identificar os efeitos de sentido de cada, suas semelhanças e distinções. O trabalho tem como objetivo compreender a circulação intersígnica das marcas de luxo e das marcas mainstream no contexto do varejo de moda. Os resultados que aqui compartilharemos é proveniente de uma pesquisa que conta com o intercruzamento dos campos da comunicação com o a antropologia visual. A comunicação, mas precisamente, a publicitária, entrará como sua retórica de consumo por meio dos signos que se relacionam para uma produção de sentido eficaz. Já a antropologia visual ficará por conta da pesquisa fotoetnográfica que nos auxiliará no mapeamento e narrativa do consumo de luxo e do mainstream. Como método de análise nos apoiaremos na Semiótica desenvolvida por Charles Sanders Peirce. O registro fotoetnográfico foi realizado nas cidades de: São Paulo, Fortaleza, Brasília e Florianópolis e buscamos fotografar as vitrines das marcas e luxo e das marcas mainstream que nos possibilitem um estudo sobre a circulação intersígnica entre as mais variadas vitrines. Este trabalho não terá como pretensão a análise de recepção por parte dos consumidores, ficaremos restritos só aos possíveis efeitos de sentido que as marcas se utilizam para ampliar seu escopo imagético e identitário. / ephemeral and only try to satisfy a small part of our constant search for completeness. Publicity is inserted in this context when expressing the values in force in society and also in mediating the possibilities of insertion of the consumers in the most diverse niches and social strata. Trademarks seek to express values, identities and images of products / services through actions and manifestations. Such actions and manifestations may come in a variety of ways, that is, in print media advertisements, or in the form of audiovisuals; Merchandising actions such as shop windows, gondolas, among others and whose purpose is to generate positive bonds, pleasant associations and lead the potential consumer to the act and usufruct of the good or service. The present work results from a research between the luxury brands in comparison of the mainstream brands, a deepening intersígnico looking for to identify the effects of sense of each, its similarities and distinctions. The work aims to understand the intersection of luxury brands and mainstream brands in the context of fashion retail. The results we share here come from a research that relies on the cross-linking of the fields of communication with visual anthropology. Communication, but precisely the advertising, will enter as its rhetoric of consumption through the signs that relate to a production of effective meaning. The visual anthropology will be based on the photoetnographic research that will help us in the mapping and narrative of the consumption of luxury and the mainstream. As a method of analysis we will rely on the Semiotics developed by Charles Sanders Peirce. Photoetnographic registration was carried out in the cities of: São Paulo, Fortaleza, Brasília and Florianópolis, and we sought to photograph the showcases of the brands and luxury and of the mainstream brands that allow us to study the intersection circulation among the most varied shop windows. This work will not have as a pretension the analysis of reception by consumers, we will be restricted only to the possible effects of meaning that the brands are used to expand their image and identity scope.