12 resultados para TRANSFER CATALYTIC-SYSTEMS


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Dissertação para obtenção do Grau de Mestre em Bioorgânica

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J Biol Inorg Chem (2010) 15:967–976 DOI 10.1007/s00775-010-0658-6

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Journal of Electroanalytical Chemistry 541 (2003) 153-162

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IEE Proceedings - Vision, Image, and Signal Processing, Vol. 147, nº 1

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IEE Proceedings - Vision, Image, and Signal Processing, Vol. 147, nº 1

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Dissertation presented to obtain a Ph.D. degree in Sciences of Engineering and Technology, Cell Technology, at the Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

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Biochemistry, 2011, 50 (20), pp 4251–4262 DOI: 10.1021/bi101605p

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Dissertation presented to obtain the Master Degree in Molecular Genetics and Biomedicine

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Dissertação para obtenção do Grau de Doutor em Engenharia Física

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Exosomes are small membrane vesicles secreted by most cell types, either normal or malignant and are found in most body fluids such as saliva, plasma and breast milk. In the past decade, the interest in these vesicles has been growing more and more since it was found that besides their beneficial functions such as the removal of cellular debris and unnecessary proteins during cell maturation process, they can also interact with other cells and transfer information between them, thus helping diseases like cancer to progress. The present work intended to use gold nanoparticles as vehicles for gene silencing in an attempt to reduce the tumor-derived exosome secretion, regulated by Rab27a protein, and also aimed to compare the exosome secretion between two breast cell lines, MCF7 and MDA. Changes in RAB27A gene expression were measured by Real-time Quantitative PCR and it was revealed a decreased in RAB27A gene expression, as expected. Exosomes were isolated and purified by two different methods, ultracentrifugation and the commercial kit ExoQuick™ Solution, and further characterized using Western Blot analysis. ExoQuick™ Solution was proven to be the most efficient method for exosome isolation and it was revealed that MDA cells secrete more exosomes. Furthermore, the isolated MCF7-derived exosomes were placed together with a normal bronchial/tracheal epithelial cell line (BTEC) for an additional assay, which aimed to observe the uptake of exosomes by other cells and the exosomes’ capability of promoting cell-cell communication. This observation was made based on alterations in the expression levels of c-Myc and miR-21 genes and the fact that they both have an increased expression in BTEC cells incubated with tumor-derived exosomes when compared to control cells (without incubation with the exosomes) lead us to the conclusion that the exosome uptake and exchange of information between the exosomes and the normal cells did occurred.