22 resultados para rodamina WT
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia Química e Bioquímica
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Conservação e Restauro,Área de especialização Cerâmica e Vidro
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Dissertação apresentada para a obtenção do Grau de Doutor em Conservação e Restauro, especialidade Ciências da Conservação, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia
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Mestrado integrado em Engenharia Química e Bioquímica
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Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquimica
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Dissertação para obtenção do Grau de Mestre em Biotecnologia
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Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau Mestre em Biotecnologia
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Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
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Dissertation presented in partial fulfilment of the Requirements for the Degree of Master in Biotechnology
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A thesis submitted in fulfillment of the requirements for the degree of the Masters in Molecular Genetics and Biomedicine
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Dissertação para obtenção do Grau de Mestre em Bioquímica
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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the pro-gressive loss of motoneurons (MN). Increasing evidence points glial cells as key players for ALS onset and progression. Indeed, MN-glia signalling pathways involving either neuroprotection or inflammation are likely to be altered in ALS. We aimed to study the molecules related with glial function and/or reactivity by evaluating glial markers and hemichannels, mainly present in astrocytes. We also studied molecules involved in mi-croglia-MN dialogue (CXCR3/CCL21; CX3CR1/CX3CL1; MFG-E8), as well as proliferation (Ki-67) and inflammatory-related molecules (TLR2/4, NLRP3; IL-18) and alarming/calming signals (HMGB1/autotaxin). We used lumbar spinal cord (SC) homogenates from mice expressing a mutant human-SOD1 protein (mSOD1) at presymptomatic and late-symptomatic ALS stages. SJL (WT) mice at same ages were used as controls. We observed decreased expression of genes associated with astrocytic (GFAP and S100B) and microglial (CD11b) markers in mSOD1 at the presymptomatic phase, as well as diminished levels of gap junction components pannexin1 and connexin43 and expression of Ki-67 and decreased autotax-in. In addition, microglial-MN communication was negatively affected in mSOD1 mice as well as in-flammatory response. Interestingly, we observed astrocytic (S100B) and microglial (CD11b) reactivity, increased proliferation (Ki-67) and increased autotaxin expression in symptomatic mSOD1 mice. In-creased MN-microglial dialogue (CXCR3/CCL21; CX3CR1/CX3CL1; MFG-E8) and hemichannel activ-ity, namely connexin43 and pannexin1, were also observed in mSOD1 at the symptomatic phase, along with an elevated inflammatory response as indicated by increased levels of HMGB1 and NLRP3. Our results suggest that decreased autotaxin expression is a feature of the presymptomatic stage, and precede the network of pro-inflammatory-related symptomatic determinants, including HMGB1, CCL21, CX3CL1, and NLRP3. The identification of the molecules and signaling pathways that are dif-ferentially activated along ALS progression will contribute for a better design of therapeutic strategies for disease onset and progression.
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Polyhydroxyalkanoates (PHAs) are biosynthetic polyesters, biodegradable and biocompatible making them of great interest for industrial purposes. The use of low value substrates with mixed microbial communities (MMC) is a strategy currently used to decrease the elevated PHA production costs. PHA production process requires an important step for selection and enrichment of PHA-storing microorganisms which is usually carried out in a Sequencing Batch Reactor (SBR). The aim of this study was to optimize the PHA accumulating culture selection stage using a 2-stage Continuous Stirrer Tank Reactor (CSTR) system. The system was composed by two separate feast and famine bioreactors operated continuously, mimicking the feast and famine phases in a SBR system. Acetate was used as carbon source and biomass seed was highly enriched in Plasticicumulans acidivorans obtained from activated sludge. The system was operated under two different sets of conditions (setup 1 and 2), maintaining a system total retention time of 12 hours and an OLR of 2.25 Cmmol/L.h-1. An average PHB-content of 3.3 % wt was obtained in setup 1 and 4.8% wt in setup 2. Several other experiments were performed in order to better understand the continuous system behaviour, using biomass from the continuous system. With the fed-batch experiment a maximum of 8.1% PHB was stored and the maximum substrate uptake and specific growth rates obtained in the growth experiment (1.15 Cmol Cmol-1.h-1 and 0.53 Cmol Cmol-1.h-1) were close to the ones from continuous system (1.12 Cmol Cmol-1.h-1 and 0.59 Cmol Cmol-1.h-1). The microbial community was characterized trough microscopic visualization, Denaturing Gradient Gel Electrophoresis (DGGE) analysis and Fluorescent in situ hybridization (FISH). The last studied performed mimicked the continuous system by building up a SBR system with all the same operational conditions while adding an extra acetate dosage during the 12 h cycle, simulating the substrate passing from the feast to the famine reactors under continuous operation. It was shown that possibly the continuous system was not able to efficiently select for PHB storing organisms under the operational conditions imposed, although the selected culture was capable of consuming the substrate and grow fast. This main conclusion might have resulted from two major factors affecting the system performance: the ammonium concentration in the Feast reactor and the amount of substrate leaching from the Feast to the Famine reactor.
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RESUMO - Objectivos Um dos problemas mais comuns nos sistemas de saúde e particularmente naqueles que têm uma estrutura predominantemente pública são os elevados Tempos Espera (TE) para cirurgia. Este projecto tem como principal objectivo identificar a importância relativa em Portugal das determinantes do TE cirúrgico. Metodologia Para clarificar essa importância, aplicou-se um questionário desenvolvido pelo Fraser Institute, utilizado para a análise anual dos TE no Canadá, recorrendo a um painel de peritos e através da realização da técnica Delphi, procurou-se consensualizar quais as determinantes das Listas Espera (LE), mais importantes. Para ilustrar a diversidade de perspectivas, recorreu-se também à análise de trabalhos realizados por inúmeras organizações, onde pudemos observar e recolher distintas abordagens, políticas e técnicas da questão das LE em países com diferentes tipos de sistemas de saúde. Resultados Os resultados obtidos revelam que os peritos consideraram a disponibilidade de tempo no Bloco Operatório (BO) e a de anestesiologistas, como as determinantes com maior impacto no aumento das LE. Conclusões Dos dados recebidos relativos ao Sistema Integrado Gestão Inscritos Cirurgia (SIGIC), bem como dos retirados de relatórios oficiais, concluímos que as LE cirúrgicas tiveram evolução positiva, nomeadamente na redução da mediana do TE da Lista Inscritos Cirurgia (LIC) (meses), no entanto, os objectivos dos Tempos Máximos Resposta Garantidos (TMRG) não estão a ser cumpridos. A análise das diferentes iniciativas e estratégias políticas para combater as LE, permitiu-nos sugerir caminhos a explorar, tendo como objectivo minorar o problema das LE: cuidados integrados, optimização da eficiência na utilização da capacidade instalada e maior aposta na cirurgia de ambulatório.