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Otimização da purificação e caracterização adicional de uma desintegrina-RGD recombinante de Bothrops alternatus e seu efeito em células endoteliais humanas (HUVEC)
(Universidade Federal de São Carlos, 2006-11-23) Pontes, Carmen Lucia Salla; Araújo, Heloísa Sobreiro Selistre de; https://lattes.cnpq.br/4065824911933203; https://lattes.cnpq.br/7468571768190030
Disintegrins are snake venom protein, of low molecular weight, rich in cysteines and RGDcontaining peptides that bind specifically to integrins αIIbβ3, α5β1, and αvβ3 expressed on platelets, endothelial and tumor cells. The biological effects of these peptides are related with biological process of cellular adhesion where receptors called integrins are presents. This dissertation describes the optimization of purification and additional characterization of a recombinant RGD-disintegrin of Bothrops alternatus, DisBa-01.. The DisBa-01 is a recombinant RGD-disintegrin, which interacts with αIIbβ3 integrin, inhibiting platelet aggregation and proliferation of endothelial and some tumor cells. In this work, a new protocol of purification was proposed for most efficient purification of the DisBa-01. The recombinant disintegrin, DisBa-01, was expressed in an optimizedbacterial system (Escherichia coli BL21(DE3) pET28a+DisBa-01) and purified by affinity chromatography. In this new protocol, the product of the purification in nickel column is submitted ion exchange chromatography. The use of the ion exchange chromatography as second step for purification increased the purity degree and the protein yield. The purified protein had its N-Terminal portion sequenced in 20 amino acids residues and its molecular mass determined by mass spectrometry. The mass spectrometry assay confirmed the mass of the DisBa-01 as 11.658 Da. The RGD-disintegrin pure had the tag of polihistidinas removed with thrombin. The cleavage of the tag of histidinas with thrombin was efficient. Polyclonal antibodies against DisBa-01 have also been produced in mice. The reactivity of these antibodies was observed through imunoblotting. These results provided one better form of obtain the pure protein as well as significant additional information for a better characterization of recombinant RGDdisintegrin, DisBa-01, which could be useful for the study of RGD-disintegrin, recombinant or native, and integrin.
Trabalhadoras do sexo e profissionais da saúde: percepções sobre os processos educativos relacionados à saúde presentes no trabalho sexual
(Universidade Federal de São Carlos, 2006-11-23) Ferreira, Flávia do Carmo; Oliveira, Maria Waldenez de; https://lattes.cnpq.br/3404769527869004; https://lattes.cnpq.br/3542452308576688
This work is focused on analising the similarities and differences of the perceptions of sex workers and public health system workers about the educative processes that is within sex work related to the sex workers health. We aim to examinate, in the point of view of both social groups, how the sex worker educates herself and others of her own class exchanging informations in the work group, with the client and with health worker seeking their perceptions about the processes which the sex worker teaches and/or learns health related knowledges, values or attitudes. We will also aim the relationship between health and sex work. This research is based on common sense definition according to Paulo Freire. It has been done with the support of the research methodology related with the participation of sex workers in interviews. Interviews are also done with health workers. We aim to contribute to destroy the stereotypes that keeps the prejudiced image of sex workers and also to help on contructing health politics that take this social group in a citizenship perpective.