Do inquérito à arquitetura regional portuguesa à investigação local aplicada: a experiência do GTAA Sotavento nos domínios da investigação sobre o património vernacular construído

Versão em português da comunicação submetida com o título “From The Survey on Regional Architecture In Portugal to the local applied research: the experience of GTAA Sotavento in the built vernacular heritage studies” à Conferência Internacional “surveys on vernacular architecture. Their significance in 20th century architectural culture” realizada no Porto (ESAP) entre 17 e 19 de Maio de 2012.

Le Comte de Monte Cristo: da literatura ao cinema

A presente dissertação discute o diálogo estabelecido entre literatura e cinema no tratamento da personagem principal – um homem traído que se vinga de forma cruel dos seus inimigos – na obra literária Le Comte de Monte-Cristo, de Alexandre Dumas, e nas três adaptações fílmicas escolhidas: Le Comte de Monte-Cristo de Robert Vernay (1943); The count of Monte Cristo de David Greene (1975) e The count of Monte Cristo de Kevin Reynolds (2002). O projecto centra-se na análise da personagem Edmond Dantès/Monte-Cristo na obra literária e na sua transposição para a obra fílmica, nos graus de adaptação e contaminação entre os diferentes textos em momentos temporais diferentes. Para isso, foram estudadas em pormenor cada uma das obras referidas, tendo sempre em vista o percurso do herói. A adaptação cinematográfica de uma obra literária é uma operação de criação de um novo texto com dinamismo e autonomia próprios, resultante de escolhas nem sempre fáceis de realizar. A grande extensão da obra literária dificultou essa tarefa, no entanto, os realizadores procuraram constituir um fluir de personagens e de situações enquadráveis num tempo ideal de visualização confortável. Todas as narrativas combinam romance e desilusão, acção e aventura, mas os temas da vingança e da traição são explorados numa perspectiva diferente. As conclusões apuradas constatam que, independentemente da época, a personagem descrita nos filmes manteve-se fiel à caracterização feita pelo seu criador Alexandre Dumas, apesar de algumas nuances necessárias para criar um objecto que garantisse reconhecimento da crítica mas, também, o sucesso comercial.

Development of a Piggybac based direct reprogramming system for derivation of integration free induced pluripotent stem cells

Induced pluripotent stem cells (iPSc) have great potential for applications in regenerative medicine, disease modeling and basic research. Several methods have been developed for their derivation. The original method of Takahashi and Yamanaka involved the use of retroviral vectors which result in insertional mutagenesis, presence in the genome of potential oncogenes and effects of residual transgene expression on differentiation bias of each particular iPSc line. Other methods have been developed, using different viral vectors (adenovirus and Sendai virus), transient plasmid transfection, mRNA transduction, protein transduction and use of small molecules. However, these methods suffer from low efficiencies; can be extremely labor intensive, or both. An additional method makes use of the piggybac transposon, which has the advantage of inserting its payload into the host genome and being perfectly excised upon re-expression of the transposon transposase. Briefly, a policistronic cassette expressing Oct4, Sox2, Klf4 and C-Myc flanked by piggybac terminal repeats is delivered to the cells along with a plasmid transiently expressing piggybac transposase. Once reprogramming occurs, the cells are re-transfected with transposase and subclones free of tranposon integrations screened for. The procedure is therefore very labor intensive, requiring multiple manipulations and successive rounds of cloning and screening. The original method for reprogramming with the the PiggyBac transposon was created by Woltjen et al in 2009 (schematized here) and describes a process with which it is possible to obtain insert-free iPSc. Insert-free iPSc enables the establishment of better cellular models of iPS and adds a new level of security to the use of these cells in regenerative medicine. Due to the fact that it was based on several low efficiency steps, the overall efficiency of the method is very low (<1%). Moreover, the stochastic transfection, integration, excision and the inexistence of an active way of selection leaves this method in need of extensive characterization and screening of the final clones. In this work we aime to develop a non-integrative iPSc derivation system in which integration and excision of the transgenes can be controlled by simple media manipulations, avoiding labor intensive and potentially mutagenic procedures. To reach our goal we developed a two vector system which is simultaneously delivered to original population of fibroblasts. The first vector, Remo I, carries the reprogramming cassette and GFP under the regulation of a constitutive promoter (CAG). The second vector, Eneas, carries the piggybac transposase associated with an estrogen receptor fragment (ERT2), regulated in a TET-OFF fashion, and its equivalent reverse trans-activator associated with a positive-negative selection cassette under a constitutive promoter. We tested its functionality in HEK 293T cells. The protocol is divided in two the following steps: 1) Obtaining acceptable transfection efficiency into human fibroblasts. 2) Testing the functionality of the construct 3) Determining the ideal concentration of DOX for repressing mPB-ERT2 expression 4) Determining the ideal concentration of TM for transposition into the genome 5) Determining the ideal Windows of no DOX/TM pulse for transposition into the genome 6) 3, 4 and 5) for transposition out of the genome 7) Determination of the ideal concentration of GCV for negative selection We successfully demonstrated that ENEAS behaved as expected in terms of DOX regulation of the expression of mPB-ERT2. We also demonstrated that by delivering the plasmid into 293T HEK cells and manipulating the levels of DOX and TM in the medium, we could obtain puromycin resistant lines. The number of puromycin resistant colonies obtained was significantly higher when DOX as absent, suggesting that the colonies resulted from transposition events. Presence of TM added an extra layer of regulation, albeit weaker. Our PCR analysis, while not a clean as would be desired, suggested that transposition was indeed occurring, although a background level of random integration could not be ruled out. Finally, our attempt to determine whether we could use GVC to select clones that had successfully mobilized PB out of the genome was unsuccessful. Unexpectedly, 293T HEK cells that had been transfected with ENEAS and selected for puromycin resistance were insensitive to GCV.

Du roman au théâtre: adaptations théâtrales dans la dramaturgie franco-portugaise au XIXe siècle

L’adaptation de romans à la scène au XIXe siècle dans la dramaturgie franco-portugaise passe par l’étude des relations et des contaminations intergénériques et interculturelles qui s’établissent entre les genres romanesque et dramatique dans les deux cultures. Pour ce faire, nous nous sommes consacrés dans une première partie aux questions théoriques qu’un tel exercice soulève. Effectivement, nous ne pourrions pas prétendre étudier et cerner un tel phénomène littéraire et artistique sans un abordage pluridimensionnel qui nous mène vers son approche à la lumière des études de Traduction et de Réception, d’un côté et, de l’autre, des études sociologiques. Questionner les fondements théoriques de l’adaptation théâtrale, sa production et sa circulation dans les deux pays supposait, pour nous, dès le départ, délimiter non seulement l’horizon d’attente d’une telle pratique artistique mais aussi sa place et son impact dans le champ littéraire et le polysystème culturel de l’époque. En contrepoint, nous nous sommes ensuite intéressés à la question de l’adaptation théâtrale et des contact zones, cette fois en passant par la problématique de la transculturation qu’engendre le contact de deux littératures différentes, s’affrontant dans le champ culturel et littéraire d’arrivée. Dans une deuxième partie, nous nous sommes concentrés sur la contextualisation historico-générique de l’adaptation théâtrale. Le recours à la transposition scénique de romans révèle un désir de gloire au niveau financier et artistique. La pratique est courante, voire généralisée dans le champ littéraire français du XIXe siècle. Parmi les adeptes de l’exercice de transposition, Dumas père est certainement l’auteur le plus prolifique. En outre, la transmodalisation générique s’insère dans un mouvement de création collective, puisque la collaboration entre auteurs dramatiques est très répandue. Par la suite, nous nous sommes intéressés à la genèse des textes à travers la transmodalisation du roman au théâtre : du roman-feuilleton au roman naturaliste. Nous avons constaté que le genre romanesque comme le genre dramatique sont empreints de contaminations intergénériques qui passent par la théâtralisation du roman puis par la romanisation du théâtre. Ainsi, l’adaptation apparaît comme la concrétisation de la fusion des genres romanesque et dramatique qui donnent lieu à un genre hybride. L’adaptation est alors le résultat d’une série de procédés et de techniques : la transmodalisation du roman s’adapte aux nécessités génériques du théâtre. De la sorte, nous avons distingué cinq types d’adaptations : l’adaptation fidèle, l’adaptation partielle, l’adaptation libre, l’adaptation pastichée et l’adaptation de l’adaptation. Dans une troisième partie, nous avons abordé le phénomène de l’adaptation dans le contexte littéraire portugais au XIXe siècle. À travers la présence accrue du théâtre français sur les scènes lisboètes, nous avons observé les rouages de la transposition d’un modèle étranger qui peut passer par différents processus, comme la traduction ou l’appropriation par l’adaptation théâtrale. Cela nous a amené à présenter cinq modèles d’adaptations théâtrales issues d’hypotextes français : l’adaptation française, la traduction fidèle de l’adaptation française, la traduction libre de l’adaptation française, l’adaptation à la couleur locale de l’adaptation française et l’adaptation portugaise d’un roman français. Nous avons également présenté quelques exemples de cas concrets, notamment Le Comte de Monte-Cristo adapté au répertoire lisboète. L’étude de ce phénomène culturel nous a permis de mieux cerner son impact sur le champ théâtral lisboète du XIXe siècle, dans la mesure où il contribue à la transculturation de la dramaturgie portugaise de l’époque.

The activity of mouse Cerberus like 2 during cardiogenesis - genetic and morphogenetic studies

Cardiogenesis is a delicate and complex process that requires the coordination of an intricate network of pathways and the different cell types. Therefore, understanding heart development at the morphogenetic level is an essential requirement to uncover the causes of congenital heart disease and to provide insight for disease therapies. Mouse Cerberus like 2 (Cerl2) has been defined as a Nodal antagonist in the node with an important role in the Left-Right (L/R) axis establishment, at the early embryonic development. As expected, Cerl2 knockout mice (Cerl2-/-) showed multiple laterality defects with associated cardiac failure. In order to identify the endogenous role of Cerl2 during heart formation independent of its described functions in the node, we accurately analyzed animals where laterality defects were not present. We thereby unravel the consequences of Cerl2 lossof- function in the heart, namely increased left ventricular thickness due to hyperplasia of cardiomyocytes and de-regulated expression of cardiac genes. Furthermore, the Cerl2 mutant neonates present impaired cardiac function. Once that the cardiac expression of Cerl2 is mostly observed in the left ventricle until around midgestration, this result suggest a specific regulatory role of Cerl2 during the formation of the left ventricular myoarchitecture. Here, we present two possible molecular mechanisms underlying the cardiac Cerl2 function, the regulation of Cerl2 antagonist in activation of the TGFßs/Nodal/Activin/Smad2 signaling identified by increased Smad2 phosphorilation in Cerl2-/- hearts and the negative feedback between Cerl2 and Wnt/ß-catenin signaling in heart formation. In this work and since embryonic stem cells derived from 129 mice strain is extensively used to produce targeted mutants, we also present echocardiographic reference values to progressive use of juveniles and young adult 129/Sv strain in cardiac studies. In addition, we investigate the cardiac physiology of the surviving Cerl2 mutants in 129/Sv background over time through a follow-up study using echocardiographic analysis. Our results revealed that Cerl2-/- mice are able to improve and maintain the diastolic and most of systolic cardiac physiologic parameters as analyzed until young adult age. Since Cerl2 is no longer expressed in the postnatal heart, we suggest that an intrinsic and compensatory mechanism of adaptation may be active for recovering the decreased cardiac function found in Cerl2 mutant neonates. Altogether, these data highlight the role of Cerl2 during embryonic heart development in mice. Furthermore, we also suggest that Cerl2-/- may be an interesting model to uncover the molecular, cellular and physiological mechanisms behind the improvement of the cardiac function, contributing to the development of therapeutic approaches to treat heart failures.

Um modelo para a melhoria da qualidade dos recursos humanos das autarquias locais

Dissertação de Mestrado, Gestão Empresarial, Faculdade de Economia, Universidade do Algarve, 2004