Since its origin scenography has been situated among two arts (the visual art and the performing art). Also the visual art itself has been since the 60's of the 20th century in a similar position: experiences of living through as a part of installation (environment since 50's) or focusing more on the action itself (happening, event, performance). In my thesis I mention such examples, when both of these creative worlds cross each other on borders. The themes memory and place included in their works may consist at least of one of these two main levels: 1.) art work with place and its specifics 2.) art work established on personal stories or sharing the experience The most important for me are phenomenon of authenticity and personal experience, (co)fealing and contextuality of the place. The buildings and installations become scenes and scenographies at the same time. The happenings, events, performances become the story. But these art approaches are just shortly mentioned in my thesis, working more like roots, context with the past in contemporary works. I prefer projects intensively close to life to the artificial works. Whether it is about high experience level continuing to the intimate dialogue or sharing the big stories, memory and places, it is done in the context of time-based media, mental maps, site specific, memory, space/site, path, memories and someone who reads (in) it and (re)interpretates that. In my author diploma art project I work with a phenomenon of Being on a visit. Which are the differences and when does it come to coherence among the two key moments — being (still) on a visit and to (already) feel like at home? I´m searching for way how to reflect the place/site, using memory not only of that place/site, but also of the people who knew it — and transforming their interpretations from different personal stories or deformations of the reality (propaganda, idealisation). I focus to make a new environment/landscape, transported to another context. In my art work I am trying to use almost all tools for portraiting the reality. Painting, photography, video, installation, sound, environment, drawing, projection, text, food, performance, dance, ritual and me as the main creator of the dialogue with the visitors.

The human immunodeficiency virus is presently one of the most significant global health issues to date, with a disease burden that encumbers developing and developed nations alike. Although current antiretroviral therapy can help patients maintain undetectable levels of the virus throughout their bodies, once the treatment is ceased, the virus will rebound and disease progression continues. Thus, modalities to; 1- stop HIV transmission and spread, or 2- eradicate the virus once it is acquired are both urgently needed. In this project, we seek to evaluate and understand the impact of a candidate vaccine therapy that targets the HIV protease cleavage sites (PCS) on viral fitness. Vaccination with this modality in a monkey model induces mutations at virus regions that are intolerant to change, presumably affecting the "fitness" of viral strains recovered from vaccines. Preliminary results of the study show that in the vaccine group (n=11), a disruption to one or more of the HIV protease cleavage sites results in improved maintenance of CD4+ T cells compared to unvaccinated controls (n=5). Furthermore, a correlation between the percentage of PCS mutations and reductions in viral load were seen. Our data indicate that the most common sites of mutation occur at two cleavage regions PCS2 and PCS12. We used site directed mutagenesis to introduce multiple PCS mutations into infectious clones of SIV. Our ongoing studies are evaluating the viral fitness of the SIV mutants in a cell lines and PBMC using competitive viral fitness assays. The data from these studies will help inform in the areas of vaccine and therapy development for HIV-1.

Retinoblastoma Binding Protein 6 (RBBP6) is a 200 KDa protein shown to play a role in 3'- polyadenylation of mRNA transcripts, as well as to function as an E3 ligase catalysing ubiquitination of cancer-associated proteins. RBBP6 has been previously reported to localise to nuclear speckles, which are thought to play a role in mRNA splicing, presumably as a result of its RS domain, which is known to target mRNA splicing factors to nuclear speckles. However recent studies in our laboratory have shown that isoform 3 of RBBP6, consisting mainly of the DWNN domain, also localises to speckles in resting cells, but more strongly in cells subjected to various stresses, suggesting that the DWNN domain may be the speckle-targeting domain. / Magister Scientiae - MSc (Biotechnology)

The diploma thesis The summer project In the cellar of the Theater Continuo is about my experience at the summer workshop organized by the Continuo Theatre concretely on site-specific project In the cellar. At first I write in a style of diary and later on I describe particular approaches of work, exercises and creating of the performance itself. The final section is devoted to the author's own project Coco.on, which was created as part of a school project Left on my own. This creation was based on the experience gained from the summer workshop.

This practice-based research examines the developments in site-specific art since 1960's. Its purpose is to critically explore the influence of patronage and the role of the artist on practices within this field and it has been achieved by adopting a binary strategy, which combines "research into art" with "research through art." (Frayling 1993:5). This has involved the construction of an individually tailored methodological framework consisting of a textual examination of historical case-studies conducted in conjunction with the organisation and running of practical projects (the BT/Cellnet Farnham Library Garden & Picnic projects). Devised both as site-specific ventures and as case-studies, these projects broadly encompass three distinctive historical modes of site-specific art - Formal/Object-based, Community-based and Performative - which have successively emerged to predominance over the past fifty years or therabouts. The aim was to produce empirical comparisons in order to reflexively investigate the ways in which patronage, coupled with the role played by the artist impacted on the form and content of site-specific art.

Os avanços nas Tecnologias de Informação, em especial a Internet, e a sua aplicação para fins comerciais, trouxe transformações para as empresas, no que se refere ao relacionamento com clientes, fornecedores e parceiros, e ao seu ambiente interno. A forma mais elementar que evidencia a conexão das empresas com essa tecnologia é a presença na rede, através de um site1. Este trabalho explora o papel da implementação de um site de uma S.A. na explicação da variação dos preços das suas ações no mercado de capitais brasileiro. O presente estudo tem por objetivo investigar se o mercado precifica a iniciativa das empresas em ingressarem no mundo virtual através da criação de um site próprio. Para uma amostra de 36 empresas brasileiras de capital aberto com ações negociadas em bolsa de valores, verificou-se, através de um estudo de evento, que a implementação do site de uma S.A. não afetou o preço de suas ações na data do evento e ao redor desta, para um período de doze meses, o que implica a aceitação da hipótese nula. Entretanto, quando realizada a segmentação da amostra por ano de lançamento do site2 e por versão do lançamento3, os resultados diferenciam-se daqueles da análise da amostra completa. Com relação aos resultados provenientes da segmentação por ano de lançamento, foram encontrados retornos anormais negativos significativos ao nível de 10% para o ano de 1995, retornos positivos significativos ao nível de 5% para o ano de 1997 e retornos positivos significativos ao nível de 10% para o ano de 2000. Para os demais anos, não foram encontrados retornos anormais significativos. Quanto aos resultados da segmentação por versão implementada, a primeira versão do site para estas empresas não resultou na existência de retornos anormais significativos, o que implica a aceitação da hipótese nula. Para as empresas cuja implementação da 2° versão foi analisada, observou-se a existência de retornos anormais negativos ao nível de significância de 10% na data +1, correspondente mês um da janela do evento.

Introdução: A profilaxia antimicrobiana cirúrgica tem se demonstrado como uma das mais importantes medidas para redução das infecções de sítio cirúrgico. O momento da administração dessa profilaxia é essencial para atingir concentrações terapêuticas nos tecidos durante o procedimento cirúrgico, condição freqüentemente não observada. O objetivo deste estudo foi caracterizar o lapso de tempo transcorrido entre a profilaxia antimicrobiana e o início da incisão cirúrgica. Métodos: Amostra de procedimentos cirúrgicos eletivos realizados em um hospital geral acompanhada mediante observação direta do momento de administração da antibioticoprofilaxia e da incisão. Resultados: Foram analisados 130 procedimentos. A maioria dos pacientes da amostra era do sexo feminino (64,6%). Em 60% dos procedimentos avaliados, os pacientes internaram no mesmo dia da cirurgia e o antimicrobiano mais utilizado foi cefazolina (91,5%). O antimicrobiano profilático foi administrado com uma mediana de 8 minutos antes da incisão, sendo que em 18 (13,8%) procedimentos foi aplicado num tempo maior que 30 min e em 37 (28,5%), foi administrado após a incisão. Em apenas 13,8% a profilaxia foi administrada conforme o protocolo da instituição. As especialidades cujos procedimentos cirúrgicos apresentaram menor freqüência de adequação do horário de administração foram cirurgias plástica (4,5%), urologia (6,7) e vascular (9,1%). Conclusão: Apesar de consagrada, a administração da profilaxia em tempo adequado ainda permanece longe do ideal no nosso meio. As instituições devem adotar medidas eficazes para minimizar a inadequação desta medida. / Introduction: Antimicrobial surgical prophylaxis has been demonstrated as one of the most important steps to reduce surgical site infections. The timing of administration of prophylaxis is essential to achieve therapeutic concentrations in tissues during the surgical procedure, which is frequently not observed. The aim of this study was to characterize the lapse of time between antimicrobial prophylaxis and beginning of surgical incision. Methods: A sample of elective surgeries performed in a general hospital accompanied by direct observation of the timing of administration of antibiotic prophylaxis and the incision. Results: We analyzed 130 procedures. Most of the patients were female (64.6%). In 60% of the procedures evaluated, patients hospitalized on the same day of surgery and antimicrobial more used was cefazolin (91.5%). The antibiotic prophylaxis was administered with a median of 8 minutes before incision, and in 18 (13.8%) procedures were applied to a time longer than 30 min and 37 (28.5%) was administered after the incision. In only 13.8% prophylaxis was administered according to institutional protocol. Whose specialty surgical procedures had lower rates of appropriateness of administration time were plastic surgery (4.5%), urology (6.7) and vascular (9.1%). Conclusion: Although established, the administration of prophylaxis in a timely manner remains far from ideal in our midst. Institutions should adopt effective measures to minimize the inadequacy of this measure.

Glucansucrases são enzimas que atuam em reações de síntese de polissacarídeos e oligossacarídeos. Para que esses biocatalisadores sejam aplicados em escala industrial, é desejável ótimas estabilidades térmica e operacional, o que pode ser alcançado com a imobilização de enzimas. Como alternativa aos suportes sólidos amplamente estudados, está a quitosana, polímero que não apresenta toxicidade e possui alta biocompatibilidade e alta afinidade com proteínas. Outra possibilidade promissora na imobilização de enzimas, é a síntese dos agregados enzimáticos entrecruzados (CLEAs), os quais apresentam alta atividade catalítica e alta estabilidade. Contudo, uma peculiaridade das glucansucrases quando produzidas em meio contendo sacarose é a camada de polímero que as envolve, e que bloqueia o acesso aos grupos reativos na superfície da proteína. No caso da expressão heteróloga das glucansucrases em Escherichia coli essa dificuldade pode ser contornada. Além disso, o uso da mutagênese sítio-dirigida pode proporcionar modificações de aminoácidos na superfície da enzima, tais como os resíduos Lys, Cys, His, com o intuito de que melhorias na imobilização sejam alcançadas. Sendo assim, na primeira etapa desse trabalho, uma extensa discussão é apresentada em relação às metodologias de imobilização de dextransucrase encontradas na literatura. A seguir, estudos referentes à imobilização da dextransucrase de Leuconostoc mesenteroides B-512 F em esferas de quitosana ativadas com glutaraldeído foram realizados. Esse imobilizado apresentou alta atividade catalítica (197 U/g) quando utilizada a carga de proteína de 400 mg/g de suporte. Além disso, observou-se que a imobilização covalente e os açúcares maltose e glicose promoveram proteção à enzima em temperaturas de 40 ºC e 50 ºC. Na etapa seguinte, a produção e a caracterização de CLEAs de dextransucrase de L. mesenteroides B-512 F foram investigados. Demonstrou-se que o tratamento com a dextranase foi essencial para a imobilização da glucansucrase e que o isopropanol foi o melhor agente precipitante. Os CLEAs apresentaram pH e temperatura ótimos de 3,0 e 60 ºC, respectivamente, enquanto que a dextransucrase imobilizada nas esferas de quitosana funcionalizada com glutaraldeído apresentaram os valores de 4,5 e 20 ºC. Ambas formas imobilizadas apresentaram boa estabilidade operacional na síntese de oligossacarídeos uma vez que após 10 ciclos, 40 % de atividade residual foi observada. Por fim, estão apresentados estudos sobre a modelagem das estruturas tridimensionais e a mutagênese sítio-dirigida das glucansucrases DSR-S vardel Δ4N and ASR C-APY del. Os modelos preditos demonstraram boa qualidade e a mutagênese sítio-dirigida não promoveu perdas significativas na atividade enzimática dos mutantes. Somente o mutante DSR_S326C mostrouse inativo. Os resultados obtidos sugerem que a imobilização da dextransucrase foi satisfatória e que cada técnica possibilita diferentes características ao imobilizado. Além disso, os imobilizados foram adequados para síntese de dextrana e oligossacarídeos. / Glucansucrases are enzymes that catalyze the synthesis of polysaccharides and oligosaccharides. In order to assure continuous processing and reuse of the biocatalyst in industrial applications, enzyme immobilization techniques are required to promote good thermal and operational stabilities. Among the several solid supports for enzyme immobilization, chitosan shows interesting properties because it is non-toxic, it is biocompatible, and it has high protein affinity. Other possibility is the production of cross-linked enzyme aggregates (CLEAs), which presents high catalytic activity and good stability. However, glucansucrases have a particularity when produced in sucrose medium, since a polymer layer surrounds the protein, blocking the access to reactive groups on the enzyme surface. To overcome this problem, it is possible to make the heterologous production of glucansucrases in Escherichia coli. Likewise, the site-directed mutagenesis may promote changes in the amino acids located on the surface to improve immobilization parameters. Therefore, this work aimed to discuss the several techniques applied for dextransucrase immobilization, and to design new immobilized biocatalysts. In a first step, it is presented a review about the distinct immobilization methodologies for dextransucrase. In a second study, an investigation about dextransucrase from Leuconostoc mesenteroides B-512 F immobilized on glutaraldehyde-activated chitosan particles was carried out. The novel immobilized biocatalyst showed 197 U/g (400 mg/g dried support) of catalytic activity. The covalent immobilization promoted protection against enzyme damages at 40 ºC and 50 ºC, whereas maltose and glucose acted as stabilizers. Furthermore, it was studied the production and characterization of CLEAs dextransucrase from L. mesenteroides B-512 F. It was demonstrated that dextranase treatment was crucial for immobilization. Isopropanol was chosen as the best precipitant agent. CLEAs presented optimal pH and temperature of 3.0 and 60 ºC, respectively, whereas it was found values of 4.5 e 20 ºC for dextransucrase immobilized on glutaraldehyde-activated chitosan particles. Both immobilized biocatalysts showed good operational stability in the oligosaccharides synthesis, exhibiting 40 % of residual activity after 10 cycles. Finally, the study concerning the homology modeling and site-directed mutagenesis of glucansucrases DSR-S vardel Δ4N and ASR C-APY del is presented. The predicted models showed good quality and it has been demonstrated that the site-directed mutagenesis did not promote significant losses in the variant enzyme activities. Only one mutant (DSR_S326C) had shown no dextransucrase activity. The results obtained in this work suggest that the immobilization of dextransucrase was satisfactory, also showing that each technique promotes different characteristics to the immobilized biocatalyst. Besides, these immobilized enzymes were feasible for the synthesis of dextran and oligosaccharides.

Este trabalho se situa na fronteira das áreas de Comunicação, fotografia e estudos do imaginário. Partimos do pressuposto de que as fotografias produzidas a partir do aplicativo Instagram e das câmeras da marca LOMO atingiram grande popularidade em razão de sua semelhança estética. Para concretizar este trabalho, fizemos uma exaustiva análise dessas duas ferramentas: seus modos de criação e fruição e sua relação com o tempo através de teóricos influentes das teorias do imaginário e da Comunicação. Além da pesquisa bibliográfica, nos utilizamos da mitodologia para buscar as imagens simbólicas recorrentes e regimes do imaginário que guiam essas ferramentas e suas constelações de imagens. Descobrimos que a semelhança estética entre as duas ferramentas seria apenas uma potencialidade, uma vez que os usuários do Instagram não têm como pulsão motora a busca pela estética, mas sim o clicar- publicar-impressionar, guiados pela técnica. Dessa forma, foi possível situar o Instagram como modelo do que seria um imaginário comunicacional, guiado pelo Regime Heroico do imaginário, caracterizado pelo titanismo e pela desestetização, enquanto a lomografia se situaria na polaridade oposta, guiada pelo Regime Místico e pela busca estética, sendo uma equilibração natural do próprio imaginário. / This work is located in the border areas of Communication, photography and imaginary studies. We assume that the photographs taken from the Instagram app and the LOMO cameras reached great popularity due to its aesthetic similarity. In order to achieve this work, we did a thorough analysis of these two tools: their modes of creation and enjoyment and its relationship to time by influential theorists of the imaginary and Communication theories. In the literature, we use the mitodologia to check the recourent symbolic images and imaginary schemes that guide these tools and their constellations of images. We found that the aesthetic similarity between the two tools would be only a potentiality, since Instagram users do not have as motor drive the search for aesthetics, but the click - publish - impress, guided by technique. Thus, it was possible to locate Instagram as a model of what would be a Communicational Imaginary, guided by the imaginary Heroic Regime, characterized by Titanism and the deaesthetization while lomography would stand in the opposite polarity, guided by Mystic Regime and the aesthetic pursuit, and a natural balancing of the imaginary itself.

Carrageenans are sulfated polysaccharides found in the cell walls of red algae with 20 – 30 % of the dry weight coming from sulfate esters. The understanding of how heterotrophic bacteria desulfate and depolymerize carrageenan has become a rather arduous endeavor as there are 15 different classes of carrageenan distinguished by the degree of sulfation and the presence or absence of a unique galactose derivative, the 3,6-anhydro-D-galactose. The depolymerization of carrageenan requires the removal of the sulfate substituents, a role fulfilled by sulfatases, which hydrolyze sulfate esters playing a key role in the regulation of sulfation states that determine the function of sulfated biomolecules. Through structural, mechanistic, and sequence-based studies a highly conserved sulfate-binding motif has been identified among sulfatases; however, the molecular determinants for substrate specificity remain largely speculative. Additionally, the largest sulfatase family S1, requires a unique catalytic residue resulting from a post-translationally modified cysteine in order to be functional thus making them difficult to study in vitro. Using a strain of Pseudoalteromonas sp. PS47 isolated in the Boraston Lab I show that the depolymerization of carrageenan is dependent on the degree of sulfation and that recognition of the leaving group is the driving force behind S1 specificity. With little information on the recognition of sulfated biomolecules, the X-ray crystal structures of the three sulfatases from PS47; PsS1_19A, PsS1_19B, and PsS1_NC in complex with their biological substrates provides a deeper understanding of how carbohydrate specific sulfatases recognize their cognate substrate and how this recognition of the leaving group can be extended to other S1 sulfatase families. Furthermore, I show that an exo-acting glycoside hydrolase (PsGH42) requires desulfation of carrageenan oligosaccharides before it can hydrolyze the β-glycosidic linkage, a new specificity of family 42. This research demonstrates how carrageenan depolymerization is entirely dependent on the functionality and specificity of the sulfatases found within the carrageenan utilization locus. / Graduate / 2019-12-07