Submitted by ANA LETICIA SGAVIOLLI null (ana.lsgaviolli@sp.senac.br) on 2016-10-25T21:37:48Z No. of bitstreams: 1 Submissão - 25.10.16.pdf: 954213 bytes, checksum: f27af9c710378005f768acd829e6aae3 (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-11-01T19:55:43Z (GMT) No. of bitstreams: 1 serignolli_als_me_bot.pdf: 954213 bytes, checksum: f27af9c710378005f768acd829e6aae3 (MD5) / Made available in DSpace on 2016-11-01T19:55:43Z (GMT). No. of bitstreams: 1 serignolli_als_me_bot.pdf: 954213 bytes, checksum: f27af9c710378005f768acd829e6aae3 (MD5) Previous issue date: 2016-09-02 / A Doença Enxerto Contra o Hospedeiro (DECH) é uma das principais complicações em pacientes pós transplante de células tronco hematopoiéticas (TCTH), onde as células T do doador agridem os tecidos do receptor resultando em uma das principais causas de mortalidade e morbidade. Ocorre em cerca de metade dos pacientes que realizaram transplante de células tronco hematopoiéticas (TCTH). A pele é um dos órgãos mais acometidos. A DECH é dividida da forma aguda (DECH-a) e crônica (DECH-c), de acordo com o tempo de aparecimento dos achados clínicos e histopatológicos. A DECH de pele é classificada em graus variados, de I a IV, de acordo com o comprometimento. O presente estudo teve como objetivo avaliar a incidência de DECH-a cutânea entre os pacientes submetidos ao TCTH alogênico, aparentado, no Serviço de Transplante de Medula Óssea do Hospital Amaral Carvalho em Jaú/SP. Também foi objetivo do estudo a análise do perfil destes pacientes, a incidência e o impacto da doença na mortalidade precoce nos 100 primeiros dias pós TCTH. Os procedimentos metodológicos envolveram análise retrospectiva de dados retirados de planilhas do setor administrativo do Serviço de Transplante de Medula Óssea do Hospital Amaral Carvalho em Jaú/SP, contendo informações de 1113 pacientes que estiveram internados para a realização de transplante de medula óssea alogênico, aparentado, no período de agosto de 1996 a dezembro de 2013. Foram incluídos 582 pacientes cujas as doenças eram de linhagem mielóide - as mielopatias malignas (Leucemia Mieloide Aguda, Leucemia Mieloide Crônica, Síndrome Mielodisplásica e Doença Mieloproliferativa Crônica) para que o grupo pesquisado se tornasse homogêneo em termos de drogas e condicionamento. Esses 582 pacientes englobaram 2 grupos: pacientes que apresentaram DECH-a exclusiva de pele e pacientes que não apresentaram nenhum tipo de DECH. Todos os dados foram revisados e submetidos a análise estatística univariada e multivariada. Na análise univariada, observa-se maior sobrevida em 10 anos para os indivíduos com DECH-a de pele grau I e II (p=0.039) e para os transplantados com células de Medula Óssea (0.018). Essa significância estatística foi mantida na análise multivariada. A principal causa da mortalidade nos primeiros 100 primeiros dias do transplante foi a recidiva da doença de base, sendo a DECH aguda responsável por 6% dos óbitos nesse período. Não houve diferença significativa para os outros dados pesquisados. Conclui-se então que uma escolha adequada de doadores, além do uso de fonte de medula óssea como principal escolha para realização de transplante de medula óssea, podem diminuir a incidência de DECH-a de pele, além de aumentar a sobrevida dos pacientes transplantados. / Graft versus host disease (GVHD) is a major complication occurred in patients posttransplantation hematopoietic stem cell transplantation (HSCT), where the donor's T cells attack the recipient's tissues resulting in a major cause of mortality and morbidity.This is a recurrent and severe event, which occurs in about half of patients who underwent hematopoietic stem cell transplantation (HSCT), the skin is one of the most affected organs. GVHD is divided as acute (GVHD) and chronic (chronic GVHD), according to the time, clinical and histopathological findings. This study aimed to evaluate the incidence of GVHDskin among patients undergoing allogeneic HSCT, akin in Marrow Transplant Service Bone Amaral Carvalho Hospital, Jaú / SP, the profile of this patient and the impact of the disease in early mortality in the first 100 days after HSCT. The methodological procedures involved retrospective analysis of data from spreadsheets in the administrative sector of the Bone Marrow Transplant Service, Hospital Amaral Carvalho - Jau, containing information of 1,113 patients who were hospitalized for performing allogeneic bone marrow transplantation, related in the period August 1996 to December 2013. to equalize the analysis were included only 582 patients whose diseases were of myeloid lineage - malignant myelopathy (Acute myeloid Leukemia, Chronic myeloid Leukemia, Myelodysplastic Syndrome and Chronic myeloproliferative disease) so that the study group became homogeneous in terms of drugs and conditioning. These 582 patients were divided into 2 groups: patients with GVHDexclusive skin and patients without any GVHD. All data were reviewed and subjected to statistical analysis. In univariate analysis, there is greater survival at 10 years for patients with GVHD to skin grade I and II (p = 0.039) and the transplanted cells Bone Marrow (0018). This significance was maintained in multivariate analysis. The main cause of mortality in the first 100 early days of transplantation was the recurrence of the underlying disease, and acute GVHD responsible for 6% of deaths in this period. There was no significant difference for the other surveyed data. It was concluded that a suitable choice of donors, and the use of bone marrow source as the primary choice for bone marrow transplantation, can reduce the incidence of GVHD, the skin, and increase the survival rate of transplant patients.

Orientador: Célia Regina Nogueira / Resumo: A Leucemia Mielóide Crônica (LMC) é uma doença malígna, clonal, da célula tronco hematopoética. A descoberta do cromossomo filadélfia e subsequente identificação do gene BCR-ABL, levaram a compreensão da biologia da doença que culminou com o desenvolvimento de drogas alvo-específicas, assim como o de métodos moleculares para o monitoramento da doença. O foco atual das pesquisas em LMC está voltado para o maior entendimento dos mecanismos moleculares e epigenéticos que levam a resistência terapêutica e progressão da doença. Estudos recentes demonstram que a expressão de micro-RNAs específicos modula oncogenes e genes supressores envolvidos no desenvolvimento de neoplasias. Ao encontro a esta tendência, propomos um estudo que procurou identificar o perfil de micro-RNAs dos pacientes bons respondedores aos tratamentos de primeira linha para LMC. Avaliamos o perfil de micro-RNAs, de 41 pacientes com LMC que atingiram resposta citogenética completa (ausência do cromossomo Filadélfia) após o tratamento com inibidor de tirosinaquinase e transplante alogênico de células progenitoras hematopoéticas, por meio do sistema de micro-RNA PCR arrays (TaqMan® Human Micro-RNA Array A e B). Identificamos uma assinatura de micro-RNA distinta entre os grupos tratados, apesar de se encontrarem no mesmo patamar de resposta clínica e citogenética. Palavras-chave: Leucemia Mielóide Crônica, micro-RNA, imatinibe, transplante, qPCR array. / Abstract: Chronic Myeloid Leukemia (CML) is a malignant clonal hematopoietic stem cell disease. The discovery of the Philadelphia chromosome and subsequent identification of the gene BCR-ABL have led to understanding the biology of the disease and the development of target-specific drugs, as well as molecular methods for monitoring the disease. The current focus of research in the CML is facing the greatest understanding of the molecular and epigenetic mechanisms that lead to therapy resistance and disease progression. Recent studies show that the expression of specific micro-RNAs modulates oncogenes and tumor suppressor genes involved in cancer development.We are proposing a new study in the literature that aims to identify the profile of micro-RNAs of patients good responders to first-line treatments for CML.Evaluated the profile of micro-RNAs in 41 CML patients who achieved a complete cytogenetic response (absence of Philadelphia chromosome) after treatment with tyrosine kinase inhibitor and allogeneic bone marrow transplantation, through the micro-RNA- PCR arrays (TaqMan® Human Micro-RNA Array A e B). We identified a distinct micro-RNA signature between the treated groups, despite being on the same level of cytogenetic and clinical response.Key Words: Chronic Myeloid LeuKemia, micro-RNA, imatinib, bone marrow transplantation, qPCR array. / Mestre

Made available in DSpace on 2014-12-17T15:32:18Z (GMT). No. of bitstreams: 1 EmelineNAL.pdf: 2548007 bytes, checksum: d30f88fa3814f13a0bc3ee25b49d05fb (MD5) Previous issue date: 2010-02-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Bone marrow transplantation (BMT) is currently the best therapeutic option for patients with hematologic diseases, solid tumors or autoimmune disorders. It is characterized by intravenous infusion of hematopoietic stem cells in order to restore marrow function. However, this procedure requires concomitant immunosuppression treatment, which favors the development of certain complications, often manifested in the oral cavity. This study aimed to evaluate the incidence of oral changes in patients undergoing BMT and to correlate these results with clinical aspects related to the patients and the transplants performed. This is a prevalence study, with cross-sectional design, carried out in a BMT service at the Institute of Onco-Hematology of Natal (ION) and Natal Hospital Center. Data collection was based on questionnaires, clinical examination of the oral cavity and consultation in the medical records. The sample consisted of 51 patients undergoing BMT. After the analysis, was found a general status with good health conditions and presence of oral changes in about half of patients who composed the sample. The manifestations observed were, in decreasing order of frequency: mucositis; gingival alteration and thrombocytopenic purpura; mucosal pigmentation; lichenoid reaction and candidiasis. The oral changes were observed more frequently in cases of allogeneic TMO, in different periods post-transplant, without significant differences related to the source of cells. It was found statistically significant association between the presence of graft-versus-host disease (GVHD) and oral changes (p < 0,001). Therefore, it is concluded that there is a relatively high incidence of changes in oral cavity of patients receiving bone marrow transplantation, a fact which confirms the need to consider this site for examination, diagnosis, treatment and prognosis of possible complications of BMT / O transplante de medula ?ssea (TMO) atualmente constitui a melhor op??o terap?utica para pacientes com doen?as hematol?gicas, tumores s?lidos ou desordens autoimunes. Caracteriza-se pela infus?o intravenosa de c?lulas progenitoras hematopo?ticas com o objetivo de restabelecer a fun??o medular. No entanto, esse procedimento requer tratamento concomitante de imunossupress?o, o que favorece o desenvolvimento de determinadas complica??es, as quais freq?entemente se manifestam na cavidade oral. Este estudo objetivou avaliar a incid?ncia de altera??es orais em pacientes submetidos ao TMO e correlacionar esses resultados com aspectos cl?nicos referentes aos pacientes e aos transplantes realizados. Trata-se de um estudo de preval?ncia, com desenho do tipo seccional, realizado no servi?o de TMO do Instituto de Onco-Hematologia de Natal (ION) e Natal Hospital Center. A coleta de dados baseou-se em aplica??o de question?rio, exame cl?nico da cavidade oral e consulta de informa??es nos prontu?rios m?dicos. A amostra foi constitu?da por 51 pacientes submetidos ao TMO. Ap?s a an?lise, constatou-se quadro geral com boas condi??es de sa?de e presen?a de altera??es orais em aproximadamente metade dos pacientes que compunham a amostra. As manifesta??es observadas foram, em ordem decrescente de frequ?ncia: mucosite; altera??o gengival e p?rpura trombocitop?nica; pigmenta??o da mucosa; rea??o liquen?ide e candid?ase. As altera??es orais foram mais frequentes em casos de TMO alog?nico, em diferentes per?odos p?s-transplantes, sem diferen?a significativa quanto ? origem das c?lulas. Constatou-se associa??o estatisticamente significante entre a presen?a de doen?a do enxerto contra hospedeiro (DECH) e altera??es orais (p < 0,001). Portanto, conclui-se que h? uma incid?ncia relativamente alta de altera??es na cavidade oral de pacientes transplantados de medula ?ssea, fato que confirma a necessidade de se considerar a import?ncia desse s?tio para exame, diagn?stico, tratamento e progn?stico de poss?veis complica??es do TMO

Hematopoietic stem cells (HSCs) have the ability of both self-renewal and differentiation. After bone marrow damage, surviving host HSCs or transplanted donor HSCs are able to restore hematopoiesis and maintain it for a long time due to the self-renewal potential. HSCs reside in a specific microenvironment in the bone marrow, in stem cell niche, which supports their survival and controls their functioning. In this study, we investigated the impact of bone marrow damage induced by increasing doses of irradiation on engraftment efficiency of transplanted donor repopulating cells. Using the CD45.1/CD45.2 congenic mouse model, we developed a new approach enabling estimation of surviving HSCs in damaged hematopoietic tissue. Its principle is in measuring of the donor chimerism resulting from transplantation of a defined dose of normal congenic bone marrow cells. The transplanted donor cells contain repopulating cells, progenitors (STRCs) and HSCs (LTRCs) that give rise to blood cell production which proceeds in parallel with that present in the host hematopoietic tissue. We applied this approach to monitor spontaneous regeneration of repopulating cells, including HSCs, in mice irradiated with a sublethal dose of 6 Gy or by a lethal dose of 9 Gy and rescued by syngenic bone marrow cells. This was...

Nos dias atuais, é crescente o interesse das diversas áreas do conhecimento pela discussão da qualidade de vida das pessoas submetidas ao transplante de medula óssea. O presente estudo tem como objetivos descrever as experiências de crianças e adolescentes sobreviventes do transplante de medula óssea e apreender como esta modalidade terapêutica afetou a qualidade de suas vidas. Participaram do estudo 14 crianças/adolescentes, com idade entre 7 e 17 anos, submetidos ao TMO em um hospital-escola do interior de São Paulo. Nesta pesquisa, de natureza qualitativa, utilizou-se a entrevista como recurso para a coleta de dados, realizada no domicílio ou nos retornos ambulatoriais. Agruparam-se os dados obtidos em três temas: o passado - lembranças da doença e do tratamento; o presente - crescer e viver na condição de ser transplantado e o futuro - novos caminhos pela vida. Ao descrever a experiência da criança e do adolescente submetidos ao transplante de medula óssea, evidenciou-se o referencial da qualidade de vida, construído segundo suas próprias vivências com a doença e o tratamento. O estudo mostrou que a criança e o adolescente que sobreviveram ao transplante de medula óssea viveram seu cotidiano dentro de seus limites e possibilidades, num movimento de reestruturação e readaptação, projetando no futuro a esperança da concretização de planos e sonhos. Mediante os resultados dos dados empíricos, pode-se afirmar que, apesar das dificuldades vivenciadas pelas crianças e adolescentes frente à doença e ao TMO, os sobreviventes têm uma visão otimista de suas vidas. / Nowadays, various knowledge areas take an increasing interest in the discussion about the quality of life of people submitted to bone marrow transplantation. This study aims to describe the experiences of children and adolescents who survived this kind of transplantation and to learn how this therapy mode affected their quality of life. Study participants were 14 children/adolescents between 7 and 17 years old, submitted to BMT at a school hospital in the interior of São Paulo, Brazil. In this qualitative research, data were collected by means of interviews, which were held at the participants’ homes or when they returned to hospital. Data were grouped into three theme areas: the past – memories of the illness and treatment; the present – growing up and living as a transplant survivor and the future – new roads in life. In describing the children’s and adolescents’ experience, the quality of life reference framework was disclosed, which is constructed according to their own experiences with the disease and treatment. The study demonstrated that children and adolescents who survived bone marrow transplantation lived their daily life within their limits and possibilities, in a restructuring and readjustment movement, projecting the hope of realizing plans and dreams in the future. As a result of empirical data, it can be affirmed that, in spite of the difficulties experienced by the children and adolescents in relation to the disease and the BMT, survivors have an optimistic view on their lives.

Made available in DSpace on 2014-07-29T15:30:39Z (GMT). No. of bitstreams: 1 Dissert_mestrado_Hildene.pdf: 565864 bytes, checksum: 757a27963d59fc60689199a0ec35fa95 (MD5) Previous issue date: 2011-08-31 / Fungal infections in immunocompromised patients, especially in hospitalized individuals, which are submitted to differente types of treatment have increased in recent decades and are cause of death of these patients. The diagnoses of infection as well as the correct identification of the fungus, in addition to the in vitro susceptibility tests are important to control the disease and thus avoid death. This work was conducted to identify the species that causes candidemia in patients with hematologic malignancies and in those submitted to bone marrow transplantation from Araujo Jorge Hospital in the city of Goiânia. The main predisposing factors for the acquisition of candidiasis, the virulence factors inherent to the microorganism, as well as the in vitro susceptibility to different antifungal agents of Candida isolates were verified in this work. Clinical samples from patients were collected and cultived in brain heart infusion and Sabouraud agar dextrose and incubated at room temperature and at 36ºC. The identification of yeasts was performed by culture in cornmeal agar to check the chlamydospore production, by germ tube production in fetal bovine serum plus 1% tween 80, by inoculation onto chromogenic agar, CHROMagar and methods of assimilation of carbohydrates. The isolates were subjected to hemolytic activity and proteinase testing as well as to in vitro susceptibility testing using broth microdilution and Etest methods. Of the 410 clinical samples originated from 144 transplant patients we identified eleven yeasts, included three (3) Candida albicans, five (5) C. parapsilosis, one (1) C. tropicalis and two (2) Candida spp. All isolates of C. parapsilosis were susceptible to all antifungal agents, while two (2) isolates of C. albicans and one (1) C. tropicalis were resistant to three azole derivatives, fluconazole, itraconazole and voriconazole. Candida spp isolates did not show hemolytic activity, they were not able to secrete the enzyme proteinase either, they were resistant to fluconazole, itraconazole and amphotericin B and susceptible to caspofungin and voriconazole. In this study we can conclude that C. parapsilosis was prevalent, showing that the emergence of Candida non albicans. Besides we can conclude that in vitro susceptibility tests are of great importance for the adequate therapy to avoid the death of patients. / As infecções fúngicas em pacientes imunocomprometidos, principalmente em indivíduos hospitalizados, que são submetidos a diferentes tipos de tratamento têm aumentado nas últimas décadas, sendo causa de mortalidade entre estes pacientes. O diagnóstico da infecção bem como a identificação correta do fungo além dos testes de suscetibilidade in vitro são importantes para controlar a doença e por conseguinte evitar a morte. Neste trabalho, foi realizada a identificação das espécies causadoras de candidemia em pacientes portadores de doenças hematológicas e submetidos a transplantes de medula óssea provenientes do Hospital Araújo Jorge na cidade de Goiânia. Os principais fatores predisponentes para aquisição de candidiases, os fatores de virulência inerentes ao microorganismo bem como a suscetibilidade in vitro a diferentes antifúngicos dos isolados de Candida foram verificados para estes pacientes. Amostras clínicas dos pacientes foram coletadas e semeadas em ágar Brain Heart Infusion bifásico e ágar Sabouraud dextrose, incubadas a temperatura ambiente e a 36ºC, e observadas diariamente. A identificação das leveduras foi realizada através de cultivo em ágar cornmeal para verificar a produção de clamidoconidios, produção de tubo germinativo em soro fetal bovino acrescido de 1% de tweeen 80, inoculação em agar cromogênico, CHROMagar e por métodos de assimilação de hidratos de carbono. Os isolados de leveduras obtidos foram submetidos aos testes de suscetibilidade in vitro usando o método da microdiluição em caldo e Etest. Testes de atividade hemolítica e de proteinase foram também realizados para estes isolados. Das 410 amostras clínicas procedentes de 144 pacientes transplantados foram identificadas onze leveduras do gênero Candida, sendo três (3) Candida albicans, cinco (5) Candida parapsilosis, uma (1) Candida tropicalis e duas (2) Candida spp. Os isolados de C. parapsilosis foram suscetíveis a todos os antifúngicos, enquanto dois (2) isolados de C. albicans e o de C. tropicalis (um) mostraram-se resistentes aos três derivados azólicos, fluconazol. Itraconazol e voriconazol. As amostras de Candida não identificadas com relação a espécie foram suscetíveis ao voriconazol e a caspofungina. não apresentaram atividade hemolítica e não foram capazes de secretar a enzima proteinase. Neste trabalho pode-se concluir que C. parapsilosis foi prevalente mostrando a emergência de outras espécies não albicans como agentes de candidíase e ainda que os testes de suscetibilidade in vitro são de grande importância para que a terapia seja usada adequadamente evitando a morte dos pacientes.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-03T12:26:28Z No. of bitstreams: 2 Dissertação - Francielly Pinheiro da Silva Borges - 2016.pdf: 3852166 bytes, checksum: 70bbe6c9a7c6c0aa1e10b9e7e9d49843 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-03T12:29:55Z (GMT) No. of bitstreams: 2 Dissertação - Francielly Pinheiro da Silva Borges - 2016.pdf: 3852166 bytes, checksum: 70bbe6c9a7c6c0aa1e10b9e7e9d49843 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-03T12:29:55Z (GMT). No. of bitstreams: 2 Dissertação - Francielly Pinheiro da Silva Borges - 2016.pdf: 3852166 bytes, checksum: 70bbe6c9a7c6c0aa1e10b9e7e9d49843 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-04-15 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The Human Cytomegalovirus (HCMV) is an important cause of morbi-mortality in recipients of allogeneic hematopoietic stem cell transplantation (AloHSCT). However, there is not a consensus on which protocol to use for monitoring the infection by HCMV and, data on the frequency and clinical manifestations of the infection in this group population are quite variable among the distinct transplant centers in the world. Thus, the main objective of the present study was to proceed the monitoring of active HCMV infection in patients undergoing AloHSCT by three different methodologies: antigenemia (AGM), nested-PCR (nPCR) and real-time PCR (qPCR) and determine viral load, correlating active infection with the clinical manifestations and prognosis of patients. For this, 21 patients undergoing AloHSCT were monitored (from pre-transplant period -5 days prior to transplantation- until one year after transplantation). For HCMV detection three methodologies were used: AGM, nPCR and qPCR, and for molecular detection a comparison was made between detection of HCMV in DNA extracted from pellet (buffy coat) and serum, in a paired manner. The results showed that the active HCMV infection was detected by at least one of three methodologies in 95.2% (20/21) of patients and 45% (9/20) of these were positive in pre-transplantation period, having been observed good agreement between the results of AGM and qPCR (kappa = 0.65). Of the 20 patients positive for active HCMV infection, 85% (17/20) were positive for the three methods and only 15% (3/20) were positive for AGM and qPCR, and negative by nPCR. Regarding the type of clinical sample, molecular techniques showed higher sensitivity to the pellet over the serum. The main alteration of patients was pancytopenia and the main complication was graft-versus-host disease. Six patients died during the study period, however, it was not possible to confirm if HCMV active infection was directly associated with the cause of death. The obtained data reveal a high positivity index and the occurrence of HCMV syndrome in patients submitted to aloHSCT. We hope that the results may assist in the therapeutical measures, as well as in the methodology of choice and the type of clinical sample for detection of active HCMV infection, in order to contribute for the inclusion of HCMV monitoring is included in the routine testing of patients. / O Citomegalovírus Humano (HCMV) constitui importante causa de morbi-mortalidade em receptores de transplantes de células progenitoras hematopoiéticas do tipo alogênico (AloTCPH). Entretanto, não existe ainda um consenso sobre que protocolo utilizar para o monitoramento da infecção por HCMV e, dados sobre a frequência e manifestações clínicas da infecção neste grupo populacional são bastante variáveis entre os diferentes centros de transplante em todo o mundo. Dessa forma, o principal objetivo do presente estudo foi realizar o monitoramento da infecção ativa por HCMV em pacientes submetidos ao AloTCPH por três metodologias distintas: antigenemia (AGM), nested-PCR (nPCR) e PCR em tempo real (qPCR), bem como determinar a carga viral, correlacionando a infecção ativa ao quadro clínico e prognóstico dos pacientes. Para tanto, foram monitorados (desde o período pré-transplante -5 dias antes do transplante- até um ano após o transplante) 21 pacientes submetidos ao AloTCPH. A pesquisa de HCMV foi realizada utilizando as metodologias de AGM, nPCR e qPCR, sendo que para as técnicas moleculares, houve comparação da detecção do HCMV em DNA extraído de pellet (creme leucocitário) e soro, de forma pareada. Os resultados mostraram que a infecção ativa pelo HCMV foi detectada por pelo menos uma das três metodologias em 95,2% (20/21) dos pacientes e 45% (9/20) destes foram positivos no período pré-transplante, tendo sido observada um boa concordância entre os resultados de AGM e qPCR (kappa = 0,65). Dos 20 pacientes positivos para infecção ativa por HCMV, 85% (17/20) foram positivos pelas três metodologias e 15% (3/20) foram positivos apenas por AGM e qPCR e negativos por nPCR. Em relação ao tipo de amostra clínica, as técnicas moleculares apresentaram maior sensibilidade em amostras de pellet, em comparação ao soro. A principal alteração apresentada pelos pacientes foi a pancitopenia e a principal intercorrência, a doença do enxerto contra o hospedeiro. Seis pacientes foram a óbito durante o período de estudo, entretanto, não foi possível confirmar se a infecção ativa por HCMV estava diretamente associada à causa mortis. Os dados obtidos revelam um elevado índice de positividade e síndrome de HCMV em pacientes submetidos ao AloTCPH. Esperamos que os resultados possam auxiliar na tomada de medidas terapêuticas, bem como na escolha da melhor metodologia assim como o tipo de amostra clínica para detecção da infecção ativa por HCMV de forma a contribuir para que a pesquisa de HCMV seja incluída na rotina de exames desses pacientes.

Orientador: Maria Helena de Melo Lima / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Enfermagem / Made available in DSpace on 2018-08-25T10:33:22Z (GMT). No. of bitstreams: 1 Trevisan_DaniloDonizetti_M.pdf: 1847455 bytes, checksum: 8d96bd8d4a5dd1d09a10c926f760691d (MD5) Previous issue date: 2014 / Resumo: Introdução: No âmbito do transplante de células-tronco hematopoiéticas - TCTH, as interações medicamentosas potenciais (IMp) podem causar impacto clínico relevante no paciente. Poucos são os estudos para elucidar esse fenômeno nestes pacientes. O enfermeiro, juntamente com sua equipe de enfermagem, são os profissionais mais envolvidos, diretamente, no processo de preparo e administração de medicamentos bem como acompanhamento integral ao paciente, o que justifica a importância da realização de pesquisas sobre o tema. Objetivo: Determinar a prevalência de Interações Medicamentosas Potenciais em pacientes submetidos a transplante de células-tronco hematopoiéticas no dia da infusão das células-tronco hematopoiéticas. Método: Estudo transversal realizado na Unidade de Transplante de Medula Óssea (TMO) do Hospital de Clínicas da Universidade Estadual de Campinas. Prescrições de medicamentos de pacientes submetidos a TCTH (alogênico ou autólogo) com idade igual ou superior a 18 anos foram inclusos neste estudo. Para traçar o perfil terapêutico dos medicamentos utilizou-se a classificação Anatomical Therapeutic Chemical (ATC) da Organização Mundial da Saúde (OMS). As IMp foram analisadas por meio da base de dados Micromedex® 2.0. Para a presente pesquisa foram de interesse as interações entre medicamentos bem como nível de gravidade, nível de evidência científica, tempo de início dos efeitos e o impacto clínico potencial. Utilizou-se análise descritiva para variáveis qualitativas e quantitativas, sendo realizado frequências absoluta e relativa, cálculo de média e desvio padrão. Os dados foram analisados pelo software estatístico SAS (Statistical Analysis System) versão 9.2. Resultados: Quarenta pacientes submetidos a TCTH foram incluídos neste estudo; 33 (82,5%) pacientes foram expostos a pelo menos uma IMp maior e uma contraindicada concomitantes. A totalidade dos pacientes expostos às IMp, tiveram risco aumentado de cardiotoxicidade. A maioria das IMp foi de gravidade maior (80,9%), com início de efeito não especificado (61,9%) e com documentação boa e excelente (52,4%). Conclusão: Pacientes de TCTH são altamente expostos às IMp clinicamente significantes. A ameaça à vida foi certa, ainda que a literatura limite-se a apontar os desfechos clínicos de apenas uma dupla de medicamentos. O manejo da IMp requer ações que incluem testes bioquímicos, instalação de monitores cardíacos e realização periódica de eletrocardiograma, implantação de prescrições eletrônicas com sistema de alerta para IMp e disponibilidade de bases de dados sobre IMp. É importante considerar o risco-benefício da combinação de medicamentos / Abstract: Introduction: In the context of hematopoietic stem cell transplantation - HSCT, potential drug-drug interactions (PDDI) can cause significant clinical impact on the patient. Few studies to elucidate this phenomenon in these patients. Nurses, along with his team of nursing professionals are more involved directly in the preparation and administration of medications and integral patient, which justifies the importance of conducting research on the topic tracking process. Objective: To determine the prevalence of Potential Drug Interactions in patients undergoing transplantation of hematopoietic stem cells on the day of infusion of hematopoietic stem cells. Method: Cross-sectional study conducted at the Bone Marrow Transplantation (BMT) of the Clinical Hospital of the State University of Campinas. Drug prescriptions for patients undergoing HSCT (allogeneic or autologous) aged over 18 years were included in this study. To trace the therapeutic profile of drugs used the Anatomical Therapeutic Chemical classification (ATC) of the World Health Organization (WHO). The PDDI were analyzed using the base Micromedex ® 2.0 data. In this research of interest were the interactions between drugs and level of severity, level of evidence, time of onset of effects and potential clinical impact. Descriptive analysis was used for qualitative and quantitative variables, being held absolute and relative frequencies, calculation of mean and standard deviation. Data were analyzed using SAS (Statistical Analysis System) version 9.2 statistical software. Results: Forty patients undergoing HSCT were included in this study; 33 (82.5%) patients were exposed to at least one PDDI major and contraindicated. All patients exposed to PDDI, had an increased risk of cardiotoxicity. Most PDDI was major severe (80.9%), with onset of effect unspecified (61.9%) with excellent e good documentation (52.4%). Conclusion: Patients HSCT are highly exposed to PDDI clinically significant. The threat to life was right, although the literature is pointing out the clinical outcomes of only a couple of drugs. The management of PDDI requires actions that include biochemical tests, installation of cardiac monitors and perform periodic electrocardiograms, implementation of electronic prescriptions with warning system for PDDI and availability of databases on PDDI. It is important to consider the risk-benefit of the drug combination / Mestrado / Enfermagem e Trabalho / Mestre em Ciências da Saúde

Deficiency in β-glucuronidase activity (EC 3.2.1.31) leads to the lysosomal storage disease mucopolysaccharidosis type VII not only in humans but also in a recently discovered murine mutant, the gus^mps/gus^mps mouse. Clinical and pathologic abnormalities common to the human and mouse phenotypes include shortened life span, dwarfism, dysmorphic facial features, skeletal deformities, corneal clouding, mental retardation and abnormal lysosomal storage material in the brain and peripheral organs. In the first part of this thesis, neonatal gus^mps/gus^mps mice and their normal littermates were transplanted with syngeneic normal bone marrow. Neurological function was then evaluated with two behavioral tests: the grooming test, a developmentally regulated and genetically based activity, and the Morris water maze test, which assessed spatial learning abilities. The results of these tests indicated that the behavioral deficits in the mutant mice were not restored to normal. Treated normal mice also showed significant functional deterioration, indicating the detrimental consequence of this therapy in the neonatal period. The second part of this thesis focused on a novel approach to somatic gene therapy using microcapsules. A non-autologous fibroblast cell line engineered to secrete high levels of β-glucuronidase was enclosed in perm-selective and immuno- protective microcapsules and implanted into the peritoneal cavity of gus^mps/gus^mps mice. During the 4 weeks of therapy, the biochemical and histological abnormalities of the mutant mice had significantly improved. β-Glucuronidase activity was restored to >50% of normal in the plasma and 11.3%-65.8% in the kidney, liver and spleen. No significant activity was found in the brain. As well, the secondary elevations of other lysosomal enzymes such as β-hexosaminidase and α-galactosidase had decreased in the kidney, liver, and spleen. Urinary glycosaminoglycan content had decreased in the treated mutants indicating that the β-glucuronidase was exerting a therapeutic effect. However, after three and a half weeks of therapy, the treated mutants became severely ill and developed haemorrhagic ascites. Since normal mice treated with similar microcapsules showed no adverse effects, we hypothesized that an immune response had been generated against the foreign protein (β-glucuronidase) by the mutants, leading to the high morbidity. Thus in spite of the biochemical and histological correction observed after bone marrow transplantation and somatic cell gene therapy, the long term efficacy of these treatments needs to be further evaluated. / Thesis / Master of Science (MS)

Human cells from acute myeloid leukemia (AML) patients are frequently transplanted into immune-compromised mouse strains to provide an in vivo environment for studies on the biology of the disease. Since frequencies of leukemia re-initiating cells are low and a unique cell surface phenotype that includes all tumor re-initiating activity remains unknown, the underlying mechanisms leading to limitations in the xenotransplantation assay need to be understood and overcome to obtain robust engraftment of AML-containing samples. We report here that in the NSG xenotransplantation assay, the large majority of mononucleated cells from patients with AML fail to establish a reproducible myeloid engraftment despite high donor chimerism. Instead, donor-derived cells mainly consist of polyclonal disease-unrelated expanded co-transplanted human T lymphocytes that induce xenogeneic graft versus host disease and mask the engraftment of human AML in mice. Engraftment of mainly myeloid cell types can be enforced by the prevention of T cell expansion through the depletion of lymphocytes from the graft prior transplantation.