9 resultados para Microscopia de polarização

em SAPIENTIA - Universidade do Algarve - Portugal


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Dissertação de mest., Biologia Marinha, Faculdade de Ciências do Mar e do Ambiente, Universidade do Algarve, 2007

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Dissertação de mest., Biologia Molecular e Microbiana, Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

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A pesca e os seus avultados rendimentos marcaram decisivamente toda a História do Algarve, desde os tempos mais remotos até ao presente, com particular acento no início da política pombalina, que fez do sotavento e da foz do Guadiana o seu principal centro de polarização económica. A concentração demográfica na faixa litoral explica-se, quase exclusivamente pela riqueza e afabilidade destas águas, em cujos portos fundeavam embarcações oriundas dos mais variados quadrantes do globo.

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Epithelial tissues are essential during morphogenesis and organogenesis. During development, epithelial tissues undergo several different remodeling processes, from cell intercalation to cell change shape. An epithelial cell has a highly polarized structure, which is important to maintain tissue integrity. The mechanisms that regulate and maintain apicobasal polarity and epithelial integrity are mostly conserved among all species and in different tissues within the same organism. aPKC-PAR complex localizes in the apical domain of polarized cells, and its function is essential for apicobasal polarization and epithelial integrity. In this work we characterized two novel alleles of aPKC: a temperature sensitive allele (aPKCTS), which has a point mutation on a kinase domain, and another allele with a point mutation on a highly conserved amino acid within the PB1 domain of aPKC (aPKCPB1). Analysis of the aPKCTS mutant phenotypes, lead us to propose that during development different epithelial tissues have differential requirements of aPKC activity. More specifically, our work suggests de novo formation of adherens junctions (AJs) is particularly sensitive to sub-optimal levels of apkc activity. Analysis of the aPKCPB1 allele, suggests that aPKC is likely to have an apical structural function mostly independent of its kinase activity. Altogether our work suggests that although loss of aPKC function is associated to similar epithelial phenotypes (e.g., loss of apicobasal polarization and epithelial integrity), the requirements of aPKC activity within these tissues are nevertheless likely to vary.

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Cancer is a multistage process characterized by three stages: initiation, promotion and progression; and is one of the major killers worldwide. Oxidative stress acts as initiator in tumorigenesis; chronic inflammation promotes cancer; and apoptosis inactivation is an issue in cancer progression. In this study, it was investigated the antioxidant, antiinflammatory and antitumor properties of hexane, ether, chloroform, methanol and water extracts of five species of halophytes: A. macrostachyum, P. coronopus, J. acutus, C. edulis and A. halimus. Antioxidant activity was assessed by DPPH• and ABTS•+ methods, and the total phenolics content (TPC) was evaluated by the Folin-Ciocalteau method. The anti-inflammatory activity of the extracts was determined by the Griess method, and by evaluating the inhibition of NO production in LPS-stimulated RAW- 264.7 macrophages. The cytotoxic activity of the extracts against HepG2 and THP1 cell lines was estimated by the MTT assay, and the results obtained were further compared with the S17 non-tumor cell line. The induction of apoptosis of J. acutus ether extract was assessed by DAPI staining. The highest antioxidant activities was observed in C. edulis methanol and the J. acutus ether extracts against the DPPH• radical; and J. acutus ether and A. halimus ether extracts against the ABTS•+ radical. The methanol extracts of C. edulis and P. coronopus, and the ether extract of J. acutus revealed a high TPC. Generally the antioxidant activity had no correlation with the TPC. The A. halimus chloroform and P. coronopus hexane extracts demonstrated ability to reduce NO production in macrophages (> 50%), revealing their anti-inflammatory capacity. The ether extract of J. acutus showed high cytotoxicity against HepG2 cancer cells, with reduced cellular viability even at the lowest concentrations. This outcome was significantly lower than the obtained with the non-tumor cells (S17). This result was complemented by the induction of apoptosis.

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O presente trabalho objetiva a montagem em escala laboratorial de um grupo de 6 protótipos de Células de Combustível Microbianas (CCM) de Câmara Simples pretendendo constituir um contributo de desenvolvimento de processos anaeróbios para tratamento de águas residuais em Estações de Tratamento de Águas Residuais (ETARs) e produção simultânea de energia elétrica. Em fase preliminar procede-se à montagem de um sistema laboratorial composto por 3 conjuntos de protótipos em duplicados com 3 granulometrias de carvão ativado granular (GAC). Os princípios que fundamentam a seleção do protótipo tipo são uma câmara anódica tubular totalmente preenchida com GAC para facilitar a fixação de biofilmes e a diminuição do espaçamento entre elétrodos com a interposição de um separador para reduzir a resistência interna. A experiência laboratorial inicia-se com uma fase de aclimatação com Água residual artificial e sequencialmente com água residual recolhida na ETAR Faro-Noroeste. Na etapa final a carga orgânica da água residual é incrementada com adição de Acetato de sódio. Os ensaios decorrem em modo de fluxo contínuo. São testados e comparados três tempos de contacto do afluente com o GAC (3, 10 e 30 horas), a combinação destes com dois separadores designados por Daramic HP 200 e GF/A e sem qualquer separador. Finalmente é efetuado um teste incrementando 6 a 8 vezes a carga orgânica do afluente. O desempenho é aferido através do traçado de curvas de polarização, curvas de potência e da percentagem de remoção da Carência Química de Oxigénio (CQO). Conclui que em alguns testes as eficiências de remoção de CQO são adequadas à legislação em vigor, que o aumento e tipo de carga orgânica do afluente e a operação sem separadores incrementam a diferença de potencial e reduzem a resistência interna e que as granulometrias do GAC testadas tiveram pouco efeito na avaliação dos parâmetros considerados.

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Ocular pathologies are among the most debilitating medical conditions affecting all segments of the population. Traditional treatment options are often ineffective, and gene therapy has the potential to become an alternative approach for the treatment of several pathologies. Methacrylate polymers have been described as highly biocompatible and are successfully used in medical applications. Due to their cationic nature, these polymers can be used to form polyplexes with DNA for its delivery. This work aims to study the potential of PDMAEMA (poly(2-(N,N’-dimethylamino)ethyl methacrylate)) as a non viral gene delivery system to the retina. The first part of this work aimed to study the potential for gene delivery of a previously synthesized PDMAEMA polymer of high molecular weight (354kDa). In the second part, we synthesized by RAFT a PDMAEMA with a lower molecular weight (103.3kDa) and similarly, evaluated its ability to act as a gene delivery vehicle. PDMAEMA/DNA polyplexes were prepared at 5, 7.5, 10, 12.5 and 20 nitrogen/phosphorous (N/P) ratio for the 354kDa PDMAEMA and at 5 and 7.5 for the 103.3kDa PDMAEMA. Dynamic light scattering and zeta potential measurements confirmed the nanosize and positive charge of polyplexes for all ratios and for both polymers. Both high and low Mw PDMAEMA were able to efficiently complex and protect DNA from DNase I degradation. Their cytotoxicity was evaluated using a non-retinal cell line (HEK293) and a retinal pigment epithelium (RPE) cell line (D407). We have found that cytotoxicity of the free polymer is concentration and time dependent, as expected, and negligible for all the concentrations of the PDMAEMA-DNA polyplexes. Furthermore, for the concentrations to be used in vivo, the 354kDa PDMAEMA showed no signs of inflammation upon injection in the intravitreal space of C57BL/6 mice. The transfection efficiency, as evaluated by fluorescence microscopy and flow cytometry, showed that the D407 retinal cells were transfected by polyplexes of both high and low Mw PDMAEMA, but with varied efficiency, which was dependent on the N/P ratio. Althogether, these results suggest that PDMAEMA is a feasible candidate for non-viral gene delivery to the retina, and this work constitutes the basis of further studies to elucidate the bottleneck in transfection and further optimization of the material.

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Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

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Dissertação de Mestrado, Oncobiologia: Mecanismos Moleculares do Cancro, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015