Currently, the transplantation of islets of Langerhans is a viable means to maintain control of blood sugar levels and reduce the risk of hypoglycemia in defined populations with brittle type I diabetes mellitus or those requiring pancreatectomy. However, the process of islet isolation is highly variable and not all isolations result in islet numbers or quality suitable for transplantation. This thesis aimed to improve transplantation success through optimization and standardization of the isolation process and to identify pretransplant variables associated with early islet engraftment. A previously disregarded enzyme activity, tryptic-like activity (TLA), has been identified to influence pancreas digestion efficiency and islet isolation success in both the preclinical and clinical situations. For human pancreases, islet isolation success rates improved from 0% in the lowest TLA group to over 50% in the highest TLA groups without affecting islet quality. These findings should help standardize evaluation of enzymes for clinical islet isolation. A closed, automated, pump-made gradient system was compared to the open, manual method for islet separation. No differences were observed in expected gradient volumes, islet yields or total purities between the two methods. The pump-made gradient system successfully removed manual influences on density gradient production while fulfilling regulatory requirements for closed system processing. Islet quantification was evaluated with computer-assisted digital imaging analysis (DIA) and a semi-closed assessment system. By using the DIA system method, which measures islet purity and pellet volume instead of manual counting methods, variation in islet counts and purity reduced by almost half. By using a transplant outcome measurement of C-peptide adjusted by blood glucose and creatinine, we identified four pretransplant factors that affect early transplant outcome. Of the four factors, one was related to the organ transport time, one to function of the islets, and two to the transplanted tissue volume. When these four factors were put into a predictive model, it accounted for about 40% of the transplant outcome. The work contained in this thesis identifies and optimizes a number of critical elements related to islet isolation and transplantation protocols.

Made available in DSpace on 2014-12-17T14:46:55Z (GMT). No. of bitstreams: 1 IzauraLSF_TESE.pdf: 3734408 bytes, checksum: e9ee844724b6ab44fc29129d9cc254a0 (MD5) Previous issue date: 2013-09-09 / The transplantation of organs and tissues presents itself as an important therapeutic option, both from a medical standpoint, the social or economic. Thus, the identification of variables that can interfere in the effectiveness of organs and tissues donation for transplantation needs to be investigated adequately, because it stands before increasing index of chronic and degenerative diseases in the population, what makes the waiting list for transplantation grow disproportionately and patients come to death without the opportunity of realization the treatment due to a lack of donors. In this context, has defined as objective of this study evaluate the factors associated with the effectiveness of organs and tissues donation for transplantation. It is a evaluative research, quantitative, prospective, with longitudinal design, developed at Central of Catchment, Notification and organ donation for transplant, Organ Procurement Organization and in six accredited hospitals to collect and transplantation of organs and tissues, in Natal/RN, between august 2010 and february 2011, after the approval of the Research Ethics Committee, under No. 414/10 and CAAE 007.0.294.000-10. The probabilistic sample without replacement was composed of 65 potential donors. It was used as an instrument of data collection a structured script non-participant observation of checklist type. Data were analyzed using descriptive statistics and presented in tables, charts, graphs and figures. For this, was used Microsoft Excel 2007 and statistical program SPSS version 20.0. To check the level of significance was chosen by applying the chi-square test (&#967;2) and Mann Whitney and caselas for less than five, it is considered the Fisher exact test. It was adopted as the significance level p-value <0.05. Among the surveyed it was observed that most of the individuals were male (50,8%), in the age group 45 years (53,8%), mean age of 42,3 years, minimum 5 and maximum 73 years (? 17,32 years). Single / widowers / divorced (56,9%), with up to completed elementary school (60,0%) in the exercise of professional activity (86,2%), catholic (83,1%) and residents in metropolitan region of Natal (52,3%). Was obtained donation effectiveness of 27,7%. There was no statistical significance between structure and effectiveness of the donation, but were observed inadequacies in physical resources (36,9%), materials (30,8%), organizational structure (29,2%) and human resources (18,5%). In the process, the maintenance phase (p= 0.004), diagnosis of brain death (p= 0.032), family interview (p&#8804; 0.001) and documentation (p= 0.001) presented statistical significance with effectiveness. Thus, it is accepts the alternative hypothesis of the study, in which is evidenced that the adequacy of the factors related to structure and process is associated to effectiveness of organs and tissues donation for transplantation. In this way, the effectiveness of organ and tissue donation ends in an essential way the rapidity and accuracy with which the donation process is conducted, requiring appropriate structure, with appropriate physical and material resources and skilled human resources to optimize the reduction of time and the suffering of those waiting for an organ or tissue transplant queued in Brazil / O transplante de ?rg?os e tecidos apresenta-se como importante op??o terap?utica, tanto do ponto de vista m?dico, quanto social ou econ?mico. Assim, a identifica??o das vari?veis que podem interferir na efetividade da doa??o de ?rg?os e tecidos para transplante precisa ser investigada de maneira adequada, pois se est? diante de crescente ?ndice de doen?as cr?nicas e degenerativas na popula??o, que faz com que a lista de espera para transplante aumente desproporcionalmente e os pacientes chegam ao ?bito sem a oportunidade de realiza??o do tratamento devido ao d?ficit de doadores. Nesse contexto, definiu-se como objetivo deste estudo avaliar os fatores associados ? efetividade da doa??o de ?rg?os e tecidos para transplantes. Trata-se de pesquisa avaliativa, quantitativa, prospectiva, com delineamento longitudinal, desenvolvida na Central de Capta??o, Notifica??o e Doa??o de ?rg?os para transplantes, Organiza??o de Procura de ?rg?os e em seis unidades hospitalares credenciadas para capta??o e transplante de ?rg?os e tecidos, em Natal/RN, entre agosto de 2010 e fevereiro de 2011, ap?s a aprova??o do projeto de pesquisa pelo Comit? de ?tica em Pesquisa, CAAE 007.0.294.000-10. A amostra probabil?stica sem reposi??o foi composta de 65 potenciais doadores. Utilizou-se como instrumento de coleta de dados um roteiro estruturado de observa??o n?o participante do tipo checklist. Os dados foram analisados por meio da estat?stica descritiva e apresentados em forma de tabelas, quadros, gr?ficos e figuras. Para tanto, utilizou-se o Microsoft Excel 2007 e o Programa Estat?stico SPSS vers?o 20.0. Para verificar o n?vel de signific?ncia optou-se pela aplica??o dos testes Qui-quadrado (&#967;2) e Mann Whitney e, para as caselas inferiores a cinco, considerou-se o teste exato de Fisher. Adotou-se como n?vel de signific?ncia p-valor <0,05. Entre os pesquisados observou-se que a maior parte dos indiv?duos era do sexo masculino (50,8%), na faixa et?ria at? 45 anos (53,8%), com m?dia de idade de 42,3 anos, m?nimo de cinco e m?ximo de 73 anos (? 17,32 anos). Solteiros/vi?vos/divorciados (56,9%), com at? o ensino fundamental completo (60,0%), em exerc?cio de atividade profissional (86,2%), cat?licos (83,1%) e residentes na regi?o metropolitana de Natal (52,3%). Obteve-se efetividade de doa??o de 27,7%. N?o ocorreu signific?ncia estat?stica entre a estrutura e a efetividade da doa??o, por?m observaram-se inadequa??es nos recursos f?sicos (36,9%), materiais (30,8%), estrutura organizacional (29,2%) e recursos humanos (18,5%). No processo, as fases de manuten??o (p=0,004), diagn?stico de morte encef?lica (p=0,032), entrevista familiar (p&#8804; 0,001) e documenta??o (p= 0,001) apresentaram signific?ncia estat?stica com a efetividade. Assim, se aceita a hip?tese alternativa do estudo, na qual se evidencia que a adequa??o dos fatores relacionados ? estrutura e processo est? associada ? efetividade da doa??o de ?rg?os e tecidos para transplantes. Desse modo, a efetividade da doa??o de ?rg?os e tecidos encerra de forma imprescind?vel a rapidez e a precis?o com que o processo de doa??o se conduz, requerendo estrutura adequada, com recursos f?sicos e materiais apropriados e recursos humanos capacitados, para otimizar a diminui??o do tempo e do sofrimento daqueles que aguardam um ?rg?o ou tecido na fila do transplante no Brasil