Estudo da reversão das alteracções funcionais da actina após processo de oxidação/redução

Relatório de estágio da licenciatura, Bioquímica, Faculdade de Ciência e Tecnologia da Universidade do Algarve, 2007

Controle activo de estruturas adaptativas com sensores e actuadores piezoeléctricos

Nos últimos anos o estudo de estruturas inteligentes tem atraído vários investigadores devido às suas potenciais vantagens numa larga gama de aplicações, tais como controle de forma, supressão de vibrações, atenuação de ruído e detecção de dano. O uso de materiais “inteligentes” tal como os materiais piezoeléctricos na forma de lâminas ou “patches”, embebidas ou coladas na superfície de estruturas construídas de materiais compósitos, permite assim obter estruturas que por um lado são adaptativas e por outro revelam excelentes propriedades mecânicas, aumentando assim bastante o desempenho e a fiabilidade de sistemas estruturais. Os materiais piezoeléctricos têm a propriedade de gerar uma carga eléctrica sob a acção duma carga mecânica e o reverso, isto é, aplicando um campo eléctrico nos elementos piezoeléctricos da estrutura, esta deforma-se. Neste trabalho, é apresentado um modelo de elementos finitos, baseado na teoria clássica de placas, desenvolvido para a análise do controle activo em estática e dinâmica lineares de estruturas integrando sensores e actuadores piezoeléctrico na forma de lâminas, os quais introduzem um grau de liberdade referente ao potencial eléctrico, por cada camada piezoeléctrica do elemento finito. É utilizado método de Newmark para a solução iterativa das equações de equilíbrio. Apresentam-se os resultados obtidos em três exemplos ilustrativos.

Padrões temporais de fornecimento de Megalopas de Carcinus Maenas na Ria de Aveiro: influência dos ventos e das marés

Dissertação mest., Biologia marinha - Ecologia e conservação marinha, Universidade do Algarve, 2007

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.

Retrato(s) de D. Paio Peres Correia

Dissertação de Mestrado, Literatura Medieval Portuguesa, Faculdade de Ciências Humanas e Sociais, Universidade do Algarve , 2005

Anticoagulantes orais: velho versus novo

Dissertação de mestrado, Ciências Farmacêuticas, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Development of gene therapy to deliver the receptor-specific rhTRAIL DHER variant to induce apoptosis in activated hepatic stellate cells for the treatment of liver fibrosis

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014