Este trabalho investiga a atuação do repórter de TV, profissional que, apesar de ser o principal narrador das histórias retratadas na televisão, se encontra à margem das pesquisas em telejornalismo. O objetivo foi mapear um estilo do repórter sob uma perspectiva histórica, buscando identificar diferenças e permanências nos elementos visuais da passagem, termo que no jargão jornalístico se refere ao momento em que o repórter aparece na reportagem. Como referencial teórico, utiliza-se o conceito de habitus de Bourdieu (2011), estudos de performance e dramaturgia de Goffman (1985), Coutinho (2012) e Gutmann (2012), entre outros autores, como Vizeu (2005), Benjamin (1996), Abreu e Lima (2010) e Mota (2001). Foram analisadas 72 reportagens das primeiras décadas do telejornalismo brasileiro e 73 reportagens contemporâneas, totalizando 145 passagens. Os resultados apontam um aumento no uso de movimentos de câmera, assim como o repórter também está se movimentando mais durante a passagem. O enquadramento mais utilizado é o plano americano, mas o plano geral, em que o repórter aparece de corpo inteiro, se tornou mais usual. O tempo médio da passagem aumentou, a formalidade do figurino está sendo deixada de lado e o grafismo surge como elemento visual auxiliar do repórter. O trabalho ainda reafirma a importância do papel do repórter de TV, figura que conduz a narrativa, confere credibilidade ao relato e estabelece um elo com a audiência. / This paper investigates the performance of the TV reporter, a professional that, despite being the main narrator of the stories on television, is excluded of researches in television journalism. The goal was to map a reporter style from a historical perspective in order to identify differences and continuities in the visual elements of the piece to camera, a term that in the journalistic jargon refers to the moment the reporter appears on TV. As a theoretical framework, we use the concept of habitus of Bourdieu (2011), performance and dramaturgy studies of Goffman (1985), Coutinho (2012) and Gutmann (2012), among others, as Vizeu (2005), Benjamin (1996), Abreu e Lima (2010) and Mota (2001). For this study, we analyzed 73 reports from the first decades of Brazilian television and 72 current reports, totalizing 145 pieces to camera. The results show an increase in the use of camera movements, and the reporter is also moving more during the piece to camera. The most widely used framework is the medium shot, but the wide shot, in which the reporter appears full body, became more common. The average time the reporter is displayed on the vídeo increased, the formality of clothing is being left aside and the graphics appears as a visual element helping the reporter. The study also reaffirms the importance of the TV reporter, that leads the narrative, gives credibility to the report and establishes a link with the audience.

Este trabalho investiga a atuação do repórter de TV, profissional que, apesar de ser o principal narrador das histórias retratadas na televisão, se encontra à margem das pesquisas em telejornalismo. O objetivo foi mapear um estilo do repórter sob uma perspectiva histórica, buscando identificar diferenças e permanências nos elementos visuais da passagem, termo que no jargão jornalístico se refere ao momento em que o repórter aparece na reportagem. Como referencial teórico, utiliza-se o conceito de habitus de Bourdieu (2011), estudos de performance e dramaturgia de Goffman (1985), Coutinho (2012) e Gutmann (2012), entre outros autores, como Vizeu (2005), Benjamin (1996), Abreu e Lima (2010) e Mota (2001). Foram analisadas 72 reportagens das primeiras décadas do telejornalismo brasileiro e 73 reportagens contemporâneas, totalizando 145 passagens. Os resultados apontam um aumento no uso de movimentos de câmera, assim como o repórter também está se movimentando mais durante a passagem. O enquadramento mais utilizado é o plano americano, mas o plano geral, em que o repórter aparece de corpo inteiro, se tornou mais usual. O tempo médio da passagem aumentou, a formalidade do figurino está sendo deixada de lado e o grafismo surge como elemento visual auxiliar do repórter. O trabalho ainda reafirma a importância do papel do repórter de TV, figura que conduz a narrativa, confere credibilidade ao relato e estabelece um elo com a audiência. / This paper investigates the performance of the TV reporter, a professional that, despite being the main narrator of the stories on television, is excluded of researches in television journalism. The goal was to map a reporter style from a historical perspective in order to identify differences and continuities in the visual elements of the piece to camera, a term that in the journalistic jargon refers to the moment the reporter appears on TV. As a theoretical framework, we use the concept of habitus of Bourdieu (2011), performance and dramaturgy studies of Goffman (1985), Coutinho (2012) and Gutmann (2012), among others, as Vizeu (2005), Benjamin (1996), Abreu e Lima (2010) and Mota (2001). For this study, we analyzed 73 reports from the first decades of Brazilian television and 72 current reports, totalizing 145 pieces to camera. The results show an increase in the use of camera movements, and the reporter is also moving more during the piece to camera. The most widely used framework is the medium shot, but the wide shot, in which the reporter appears full body, became more common. The average time the reporter is displayed on the vídeo increased, the formality of clothing is being left aside and the graphics appears as a visual element helping the reporter. The study also reaffirms the importance of the TV reporter, that leads the narrative, gives credibility to the report and establishes a link with the audience.

I den här uppsatsen har jag undersökt hur tre professionella skribenter, alla reportrar på Läns-tidningen Södertälje, har upplevt att ge respons på varandras texter och få respons på sina egna. Textresponsen har omfattat de tre aspekterna formalia, språk och disposition. De frågeställningar jag ställt upp för min undersökning är: Vad har reportrarna för bakgrund och hur ser deras skrivprocess ut före textresponsarbetet? Vilka förväntningar har reportrarna inför textresponsarbetet? Hur uppfattar reportrarna textresponsarbetet? Upplever reportrarna att deras skrivprocess påverkats av textresponsarbetet och i så fall på vilket sätt? Jag har använt mig av en kvalitativ metod i min undersökning. Genom två intervjuer med varje reporter, en före och en efter textresponsarbetet, har jag fått ovanstående frågor besvarade. Min analysmetod är etnografisk, då jag observerat deltagarna under intervjuerna och därefter analyserat deras svar utifrån tidigare forskning. Intervjuerna visar att reportrarna före textresponsarbetet hade väl utvecklade sätt att lägga upp skrivarbetet. Skribenterna har angett att deras skrivprocesser inte förändrats så mycket efter textresponsarbetet, förutom att de tänker till en extra gång innan de markerar en artikel som klar. De förväntningar reportrarna hade inför responsarbetet skilde sig ganska mycket åt, allt ifrån ett allmänt utbyte av idéer och infallsvinklar till konkreta tips på hur texterna kan för-bättras. I den avslutande intervjun med var och en av de tre reportrarna kom det fram att de alla upplevde det som svårt att ge respons, av rädsla att såra de andra, men att de kände att gruppens sammansättning av kolleger gjorde det lättare att ge negativ respons. När det gäller att få respons är deltagarna nöjda med de synpunkter som kom fram, även om det ibland var tufft att få negativ respons. En av deltagarna tyckte sig ha fått obefogad kritik. Reportrarna i min undersökning är alla intresserade av att fortsätta med textrespons på arbetsplatsen.

A leishmaniose é causada por protozoários do gênero Leishmania e no Brasil, as principais espécies causadoras da leishmaniose cutânea são Leishmania (V.) braziliensis e Leishmania (L.) amazonenses. O tratamento da leishmaniose apresenta diversas dificuldades, portanto é fundamental a descoberta de novos fármacos ativos, podendo ser detectada em células cultivadas in vitro e também em animais íntegros, através da técnica de bioimageamento. Neste trabalho, propusemo-nos a produzir linhagens de L. (V.) braziliensis e L. (L) amazonenses expressoras de luciferase e caracterizar o comportamento das linhagens mutantes em testes de sensibilidade a fármacos e de infecção in vitro e in vivo. Foi confirmada a emissão de luz pelas linhagens mutantes das duas espécies de Leishmania, em promastigotas e amastigotas. O comportamento das linhagens mutantes obtidas em relação a curvas de crescimento, sensibilidade aos fármacos tamoxifeno e anfoterina B em promastigotas, perfil de infectividade e sobrevivência em macrófagos e sensibilidade de amastigotas à anfotericina B foi comparado ao comportamento das linhagens parentais, não sendo observadas diferenças significativas. Camundongos BALB/c infectados com a linhagem expressora de luciferase de L. (L.) amazonenses desenvolveram lesões comparáveis aos animais infectados com a cepa selvagem, sendo possível quantificar a carga parasitária nesses animais por bioimageamento. Os resultados obtidos neste trabalho indicam que os parasitas mutantes expressores de luciferase obtidos podem ser utilizados em testes de sensibilidade a fármacos tanto in vitro como in vivo, representando um avanço metodológico nessa área de pesquisa. / Leishmaniasis is caused by protozoan parasites in Brazil, the main causative species of cutaneous leishmaniasis are Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonenses. The treatment of leishmaniasis presents several difficulties, and the discovery of new active drugs for the treatment of leishmaniasis is therefore fundamental. The enzyme luciferase is a reporter widely used in screening tests for new drugs. This enzyme catalyzes the oxidation of luciferase in the presence of ATP emitting light that can be detected in cultured cells in vitro as well as in intact animals, using the technique of bioimaging. In this work, we sought to produce strains of L. (V.) braziliensis and L. (L) amazonenses expressing luciferase and characterize the behavior of these mutant strains in drug susceptibility tests and in in vitro and in vivo infections. Production of light was detected in mutants of both species, in all life cycle stages. Mutant strains were compared to their corresponding parental lines as to their growth pattern, infectivity and survival profile in macrophages and sensitivity to amphotericin B and tamoxifen. No significant differences were observed for these parameters. BALB/c mice infected with the luciferase expressing line of L. (L.) amazonenses developed lesions comparable to those in animals infected with the wild-type strain. The parasite load in these animals was quantified through bioimaging. The results obtained of this study indicate that the mutant parasites expressing luciferase can be used for drug susceptibility testing in vitro and in vivo, representing a methodological advance in this area of research.

Clostridium difficile is a strictly anaerobic, spore-forming bacterium that is linked to over 250,000 infections annually in the United States. One of the greatest challenges facing C. difficile research has been the lack of genetic tools. This limited repertoire is due, in part, to the anaerobic nature of C. difficile. For example, most fluorescent protein reporters require O2 for chromophore maturation. Here, we demonstrate that O2-dependent fluorescent proteins produced anaerobically can acquire fluorescence after cells are fixed with cross-linkers to preserve native patterns of protein localization. This was shown using the blue and the red codon-optimized fluorescent proteins, CFPopt and mCherryOpt, respectively. Little is known about cell division in C. difficile. Here we identify and characterize a three-gene operon encoding cell division proteins found only in C. difficile and a small number of closely related bacteria. These proteins were named MldA, MldB, and MldC, for midcell localizing division proteins. MldA is predicted to be a membrane protein with coiled-coil domains and a peptidoglycan-binding SPOR domain. MldB and MldC are predicted to be cytoplasmic proteins; MldB has two predicted coiled-coil domains, while MldC lacks obvious conserved domains or sequence motifs. Mutants of mldA or mldB had morphological defects, including loss of rod shape (a curved cell phenotype) and inefficient separation of daughter cells (a chaining phenotype). Fusions of CFPopt to MldA, MldB, and MldC revealed that all three proteins localize sharply to the division site. Mutants lacking the Mld proteins are severely attenuated for pathogenesis in a hamster model of C. difficile infection. Because all three Mld proteins are essentially unique to C. difficile, they could be exploited as targets for antibiotics that combat C. difficile without disrupting the intestinal microbiome. C. difficile pathogenesis is mediated primarily by two large exotoxins called Toxin A (TcdA) and Toxin B (TcdB). Transcription of tcdA and tcdB depends on TcdR, an alternative sigma factor for RNA polymerase. Previous studies have shown both toxins are produced upon entry into stationary phase, and that this response is mediated in part by the CodY repressor, which senses GTP and branched chain amino acids. Here we used mCherryOpt as a reporter of gene expression to visualize toxin expression at the level of individual cells. This approach led to the unexpected discovery that only a subset of cells in the population induces expression of tcdA (and tcdB under specific conditions). In other words, toxin production is a “bistable” phenotype. Further experiments indicated TcdR plays a central role in mediating bistability, while CodY makes a minor but still significant contribution to bistability. Why it is advantageous for only a subset of C. difficile cells to produce toxin is not known, but one interesting possibility is related to conflicting requirements for transmission to a new host. Some cells produce toxin to provoke diarrhea while other cells differentiate into spores that can survive exposure to air.

The bacteroides group is a subdivision in the <I>Cytophaga-Flavobacterium-Bacteroides</I> phylum. This group is as phylogenetically distinct from other Gram-negative enterics, including <I>Escherichia coli</I>, as they are from Gram-positive organisms. Furthermore, there is no cross expression between genes of <I>E. coli</I> and <I>Bacteroides</I> species. It is thought that this difference in gene expression lies in part at the level of transcription initiation and is due to the sequences within the promoter region itself. A putative consensus sequence for <I>Bacteroides</I> promoters has been published by C. Jeff Smith&#146;s research group based on alignments of the sequences upstream of certain regulated genes. However, this consensus has not been found within all putative <I>Bacteroides</I> promoters. In this study, the promoter structure and function of a strong housekeeping <I>B. thetaiotaomicron</I> 16S rRNA promoter was examined and compared to an <I>E. coli</I> 16S rRNA promoter. Our hypothesis is that there are significant differences between the promoters of these two organisms. Analysis of <I>B. thetaiotaomicron</I> sequence upstream of the 16S rRNA gene has revealed the same overall structure known for <I>E. coli</I> 16S rRNA promoters in that there are two putative promoters separated by approximately 150 bp. However, the <I>B. thetaiotaomicron</I> 16S rRNA promoter contains the proposed <I>Bacteroides</I> &#151;7 and &#151;33 consensus sequences instead of the well known <I>E. coli</I> &#151;10 and &#151;35 consensus sequences. The biological activity of the<I> B. thetaiotaomicron</I> 16S rRNA full-length promoter was confirmed using a <I>Bacteroides lux</I> reporter system. A newly designed <I>Bacteroides lux</I> reporter was used to analyze specific regions of the <I>B. thetaiotaomicron</I> 16S rRNA promoter. In addition, by pairing the <I>B. thetaiotaomicron</I> 16S rRNA promoter with an <I>E. coli</I> ribosomal binding site, and vice-versa, the improved <I>lux</I> reporter was used to further confirm that the difference in gene expression between the two species lies at the level of transcription in <I>E. coli</I>. In <I>Bacteroides</I>, however, transcription and translation may work together to create a barrier to efficient gene expression of foreign genes. </P> / Master of Science

Mycobacterial species are diverse organisms, classified into tuberculous and nontuberculous mycobacteria (NTM). Mycobacterium tuberculosis has been thoroughly investigated, but pathogenic NTM have not. The identified technology gap for studying potential antibiotic targets across mycobacteria is that there is not a tool developed for efficiently creating bacterial clones containing these genes with a reporter system to evaluate CRISPR interference (CRi) knockdowns. CRi is a quick and simple way to silence genes. In this study, Golden Gate (GG) cloning compatible Lux reporter plasmids were engineered for the efficient cloning of target genes as transcriptional fusions with luxAB, a luminescent reporter, for use with CRi. Additionally, a CRi plasmid was designed with a Giles integration site so that it could be integrated into the mycobacterial genome with the reporter plasmid, but at a different location. Based on current research, it seems that mycobacterial L,D-transpeptidase enzymes (Ldts), involved in peptidoglycan synthesis, are potential targets for the drug class known as β-lactams and should be further explored. Ldt 2 is of particular interest as research indicates that it may be involved in pathogenicity; therefore, GG cloning of M. smegmatis (Msm) Ldt 2 was performed using the designed GG plasmid. Constructing the GG plasmid (pMV306hsp+luxG13, GG pMV) as well as the CRi + Giles integration plasmid (pLJR962 + pML1357, CRi + Giles) was successful; however, the evaluation of the luminescent reporter with CRi knockdown has yet to be performed.

No description available.

No description available.

[pt] Uma frase muito comum no meio jornalístico diz que a notícia não tem hora para acontecer. Pode ser de dia, de tarde, de noite e até mesmo de madrugada. Para que nada fique de fora do radar jornalístico, muitos profissionais trabalham nos horários em que a maioria da população dorme. Os repórteres da madrugada – figuras pouco prestigiadas dentro e fora das redações, mas com grande contribuição para a prática do jornalismo diário em suas empresas – são o objeto desta pesquisa. Os objetivos são entender quais são as práticas adotadas por esses profissionais, as rotinas produtivas de seus trabalhos e as estratégias que usam para enfrentar os desafios inerentes ao horário. Na busca dessas respostas, foram realizadas pesquisa de campo com viés etnográfico e entrevistas em profundidade com jornalistas do Rio de Janeiro. O campo jornalístico, as relações de forças nele inseridas e as teorias que tentam explicar suas particularidades compõem a revisão bibliográfica, que trata ainda das etapas do processo de construção da notícia. Observamos que, na madrugada, o trabalho de apuração exige a ida do repórter à rua para descobrir informações. Em muitas empresas, profissionais acumulam funções para executar este trabalho. A madrugada é sinônimo de dificuldade: os jornalistas enfrentam limitações tanto em relação à apuração e ao trabalho jornalístico como um todo quanto em relação à segurança nas ruas. Isso contribui para um companheirismo maior entre os concorrentes. Há diferenças significativas na rotina e nas práticas dos repórteres da madrugada em relação aos que trabalham em período diurno. / [en] A common sentence between journalists says that news does not have a right time to happen. It may be in the morning, in the afternoon, at night or overnight. Many professionals have to work while most people sleep in order to find stories that could be out of newsroom eyes. Overnight reporters – journalists with low status inside and outside newsrooms, but with a major role on a newsroom daily routine – are the object of this project. The focus is to understand which practices these professionals assume, the productive routines of their works and the strategies used to face challenges related to the time. In this search, we have done field research using ethnography techniques and in-depth interviews with journalists from Rio de Janeiro. The journalistic field, the power relations inside this field and theories that try to explain how journalism works take part of this literature review, as well as the steps of making news process. We have seen that, during overnight, the process of gathering information demands work outside the newsroom. In many companies, professionals have more than one function. Overnight is a synonym to difficulty: journalists have to handle with restrictions on news verification and also on their own security. These issues enhances fellowship between competitors. There are important differences in routines and practices of overnight reporters if we compare with ones that work on daytime.