No description available.

A method to efficiently introduce supersonic drag predictions from computational fluid dynamics (CFD) calculations in a combined aerodynamic-structural optimization of a High-Speed Civil Transport (HSCT) is presented. To achieve this goal, the method must alleviate the large computational burden associated with performing CFD analyses and reduce the numerical noise present in the analyses. This is accomplished through the use of response surface (RS) methodologies, a variation of the variable-complexity modeling (VCM) technique, and coarse grained parallel computing. Variable-complexity modeling allows one to take advantage of the information gained from inexpensive lower fidelity models while maintaining the accuracy of the more expensive high fidelity methods. The utility of the method is demonstrated on HSCT design problems of five, ten, fifteen, and twenty design variables. Motivation for including CFD predictions into the HSCT optimization comes from studies detailing the differences in supersonic aerodynamic predictions from linear theory, Euler, and parabolized Navier-Stokes (PNS) calculations for HSCT configurations. The effects of these differences in integrated forces and distributed loads on the aircraft performance and structural weight are investigated. These studies indicate that CFD drag solutions are required for accurate HSCT performance and weight estimates. Response surface models are also used to provide useful information to the designer with minimal computational effort. Investigations into design trade-offs and sensitivities to certain design variables, available at the cost of evaluating a simple quadratic polynomial, are presented. In addition, a novel and effective approach to visualizing high dimensional, highly constrained design spaces is enabled through the use of RS models. <i>NOTE: An updated copy of this ETD was added in July 2012 after there were patron reports of problems with the original file.</i> / Ph. D.

Acute myeloid leukemia (AML) is an aggressive hematological malignancy. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) represents a curative treatment option for AML. Alloreactive donor T cells can produce the graft-versus-leukemia (GVL) effect which represents a major therapeutic benefit of allo-HSCT. However, evading from allogeneic immune surveillance, potentially through the downregulation of human leukocyte antigen class II (HLA-II) antigen presentation machinery, can contribute to AML relapse. Lysine-specific demethylase 1 (LSD1) is an emerging epigenetic therapeutic target in AML. The present study aims to explore whether LSD1 inhibition can enhance AML immunogenicity to promote T-cell mediated immune response. The immunological effects of a clinical-stage LSD1 inhibitor bomedemstat (IMG-7289) were examined in both human and murine AML cell models in vitro. The results demonstrated that bomedemstat treatment significantly enhanced the expression of class II transactivator (CIITA). It subsequently led to the upregulation of HLA-DR in certain human AML cell lines when stimulated by IFN-γ. Bomedemstat also markedly upregulated the expression of CD86 in all human AML cell lines tested. The study also demonstrated that bomedemstat treatment significantly increased the production of pro-inflammatory cytokines IL-12 and CXCL-10. In murine AML models, bomedemstat concurrently upregulated the expression of major histocompatibility complex class II (MHC-II) and CD86 in H9M-transformed cells without IFN-γ stimulation. This effect was not observed in MN1-transformed cells. Bomedemstat-treated H9M cells were subsequently shown to induce antigen-dependent T cell activation. Functional assays revealed that bomedemstat treatment sensitized H9M cells to antigen-dependent immune killing effect mediated by CD4+ T cells. In conclusion, the current study demonstrates both phenotypically and functionally that LSD1 inhibition by bomedemstat treatment can enhance AML immunogenicity. This is evident by the increased antigen presentation, co-stimulation and production of inflammatory cytokines. These findings suggest that LSD1 inhibition may be a relevant strategy to pursue as a maintenance therapy after allo-HSCT. / Thesis / Master of Science in Medical Sciences (MSMS) / Acute myeloid leukemia (AML) is an aggressive blood cancer characterized by the rapid growth of abnormal blood cells in the bone marrow. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an important treatment for AML that involves taking blood stem cells from a healthy donor and transplanting them into an AML patient. Meanwhile, immune cells such as T cells from the donor can help destroy leukemia cells. However, AML frequently reappears after allo-HSCT and new therapies are needed to prevent the disease from coming back. The present study investigates whether blocking a protein called lysine-specific demethylase 1 (LSD1) can increase T cells’ ability to identify and kill cancer cells. The results demonstrate that treatment with an LSD1 blocker called bomedemstat can enhance the recognition of AML cells by T cells, thereby enhancing their immune response. These findings suggest that blocking LSD1 is a promising approach to enhance the effectiveness of allo-HSCT.

Introdução: Métodos quantitativos sensíveis e acurados para monitorar o quimerismo hematopoiético após Transplante de Células-Tronco Hematopoiéticas (TCTH) são necessários para verificar o sucesso do enxerto, uma vez que os resultados influenciam a abordagem terapêutica. Método: A técnica de detecção de DNA baseada em primers inserção-específicos de marcadores genéticos do tipo InDel (Inserção/Deleção) foi avaliada quanto a sua utilidade em quantificar quimerismo em amostras de DNA em baixa concentração, em PCR em Tempo Real (qPCR). Para isso, amostras de DNA de dez pacientes submetidos a Transplante de Medula Óssea (TMO) foram analisados para quantificar a concentração de DNA residual em diferentes períodos pós-transplante. Os resultados obtidos pela InDelqPCR foram comparados com a evolução clínica descrita nos prontuários. Resultados: As quantificações do DNA residual variaram de 0,021 ng/µL a 11,71 ng/µL, correspondendo a fração de 0,065% a 40,6% do DNA total presente na amostra. Os resultados obtidos (presença ou ausência do alelo inserção) foram condizentes com a evolução clínica dos pacientes, em alguns casos, evidenciando quimerismo prévio ao relatado nos prontuários. Conclusão: Nossos dados demonstram a utilidade do método InDel-qPCR, baseada em primers inserçãoespecíficos, no monitoramento de quimerismo hematopoiético. / Introduction: Sensitive and accurate quantitative methods to monitor hematopoietic chimerism after Hematopoietic Stem Cell Transplantation (HSCT) are necessary to evaluate engraftment since the results influence a therapeutic approach. Method: DNA detection technique based on insert-specific primers for Insertion/Deletion polymorphism (InDel) was evaluated for chimerism quantification of samples with low amounts of DNA, by quantitative real-time polymerase chain reaction (qPCR). Samples of patients undergoing Bone Marrow Transplantation (BMT) collected at different post-transplantation times were analyzed to quantify the residual DNA concentration. Then, the results found using InDel-qPCR were compared to the medical records. Results: DNA quantifications ranged from 0.021 ng/?L to 11.71 ng/?L, corresponding to a fraction of 0.065% to 40.6% of the total DNA. Our results, in some cases, shows chimerism presence previously to that reported in the medical records. Conclusion: Our data demonstrate the usefulness of the InDelqPCR method based on insertion-specific primers in the monitoring of hematopoietic chimerism.

Submitted by LUCIA HELENA DE ALMEIDA TESTA null (luciatesta2011@gmail.com) on 2016-03-07T22:13:50Z No. of bitstreams: 1 Infecção viral respiratória comunitária e hospitalar em pacientes submetidos a transplante de células tronco hematopoiéticas.pdf: 1140941 bytes, checksum: 93b0313920244f35bff64660455608c7 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-09T14:00:14Z (GMT) No. of bitstreams: 1 testa_lha_me_bot.pdf: 1140941 bytes, checksum: 93b0313920244f35bff64660455608c7 (MD5) / Made available in DSpace on 2016-03-09T14:00:14Z (GMT). No. of bitstreams: 1 testa_lha_me_bot.pdf: 1140941 bytes, checksum: 93b0313920244f35bff64660455608c7 (MD5) Previous issue date: 2016-02-03 / As infecções por vírus respiratórios (VR) são causas importantes de mortalidade em pacientes submetidos a Transplante de Células-Tronco Hematopoiéticas (TCTH) especialmente no período anterior à pega do enxerto. Estas infecções também podem ser adquiridas dentro dos hospitais, possivelmente transmitida por contato com profissionais de saúde ou cuidadores infectados, ou com objetos ou superfícies contaminadas. Portanto, é importante caracterizar o tipo de transmissão para que medidas rigorosas de controle possam ser implantadas. Objetivos: Analisar os casos de infecção por vírus respiratórios (VR) nos pacientes submetidos a TCTH entre agosto de 2010 a dezembro de 2013 e caracterizar os tipos de transmissão durante esse período. Método: O presente estudo foi realizado no Hospital Amaral Carvalho de Jahu nas unidades de internação e ambulatório de TCTH e na unidade de hematologia, incluindo pacientes com diagnóstico comprovado de VR por imunofluorescência ou PCR multiplex em amostras de lavado nasal. A transmissão foi definida como hospitalar na ausência de sintomas respiratórios à admissão e diagnóstico de VR comprovado laboratorialmente após cinco dias da internação ou até cinco dias após a alta hospitalar. Resultados: Durante este período 187 pacientes tiveram 214 episódios de infecção por VR. Cento e oitenta e três (85,5%) foram considerados infecção comunitária e 31 (14,5%) episódios foram considerados infecção hospitalar, sendo que 17 (7,9 %) episódios ocorreram na unidade de TCTH e 21 (9,8 %) episódios na unidade de hematologia (p=NS). A permanência hospitalar por mais de 23 dias se associou a transmissão hospitalar (p=<0,001) e o ano de 2013 mostrou uma queda significante desse tipo de transmissão (p=0,04). O VSR foi o VR com maior frequência de progressão para pneumonia. Conclusão: Concluímos que a higienização das mãos, coleta de lavado nasal (LN) antes das internações para o transplante de células tronco hematopoiéticas (TCTH), isolamento de contato para os pacientes com vírus respiratório positivo, busca ativa de sintomas e a educação continuada para os pacientes, familiares e profissionais da saúde devem ser contínua para o controle das infecções por VR nas unidades de TCTH. A maioria dessas medidas são de baixo custo e altamente efetivas. Cuidadores, contactuantes domiciliares e profissionais de saúde devem aderir às medidas de controle para garantir a segurança dos pacientes. / Introduction: Community-acquired respiratory viruses (RV) are the most frequent etiologic agents causing acute respiratory infections (ARI) in humans. These agents have a wide antigenic range, universal distribution, affect people in all age groups, and may cause various clinical syndromes involving both the upper and lower respiratory tract. These respiratory infections are major causes of mortality in patients undergoing hematopoietic stem cell transplantation (HSCT), especially in the period prior to engraftment. These infections may also be acquired in hospitals, possibly transmitted by contact with infected health professionals or patient caregivers, or with contaminated objects or surfaces. Since 2008, a continued education program was started at the HSCT Program of Amaral Carvalho Foundation aiming to improve the control of RV transmission. Patients, caregivers, donors, family members and employees are invited to participate in the activities. Objectives: To review the cases of RV infections in patients undergoing HSCT from August 2010 to December 2013, characterize the type of transmission, if community- or hospital-acquired during this period, and determine the morbidity and mortality of RV infections. Methods: The study was conducted at the HSCT Service of Amaral Carvalho Hospital, analyzing the charts of HSCT recipients with RV infection diagnosed by immunofluorescent assay or multiplex PCR. Medical data and images from patients admitted to the HSCT and hematology wards, as well as from patients assisted at the outpatient clinic were retrospectively reviewed. Hospital transmission was defined when the interval between hospital admission and the first symptoms was more than five days, or when the interval between patient discharge and the first symptoms was up to five days. Results: During this period, 187 patients had 214 episodes of VRI. Thirty-one episodes (14.5%) were considered hospital-acquired. Rates of hospital transmission were similar between HSCT unit (7,9%) and the hematology ward (9,8%). Hospital stay for more than 23 days was associated with hospital transmission (p=0.001) and a significant decrease in this type of transmission was observed in 2013 (p=0.04). VSR was the RV with the highest frequency of progression to pneumonia (42%). Conclusion: We conclude that hand hygiene, nasal lavage collection (LN) before hospitalizations for hematopoietic stem cell transplantation (HSCT), contact isolation for patients with positive respiratory virus, active pursuit of symptoms and continuing education for patients, family and healthcare professionals should be continuous for the control of infections in HSCT VR units. Most of these policies have low cost and are highly effective. Caregivers, household contacts and health professionals must comply with the control policies to ensure the safety of patients.

Orientador: Clarisse Martins Machado / Resumo: As infecções por vírus respiratórios (VR) são causas importantes de mortalidade em pacientes submetidos a Transplante de Células-Tronco Hematopoiéticas (TCTH) especialmente no período anterior à pega do enxerto. Estas infecções também podem ser adquiridas dentro dos hospitais, possivelmente transmitida por contato com profissionais de saúde ou cuidadores infectados, ou com objetos ou superfícies contaminadas. Portanto, é importante caracterizar o tipo de transmissão para que medidas rigorosas de controle possam ser implantadas. Objetivos: Analisar os casos de infecção por vírus respiratórios (VR) nos pacientes submetidos a TCTH entre agosto de 2010 a dezembro de 2013 e caracterizar os tipos de transmissão durante esse período. Método: O presente estudo foi realizado no Hospital Amaral Carvalho de Jahu nas unidades de internação e ambulatório de TCTH e na unidade de hematologia, incluindo pacientes com diagnóstico comprovado de VR por imunofluorescência ou PCR multiplex em amostras de lavado nasal. A transmissão foi definida como hospitalar na ausência de sintomas respiratórios à admissão e diagnóstico de VR comprovado laboratorialmente após cinco dias da internação ou até cinco dias após a alta hospitalar. Resultados: Durante este período 187 pacientes tiveram 214 episódios de infecção por VR. Cento e oitenta e três (85,5%) foram considerados infecção comunitária e 31 (14,5%) episódios foram considerados infecção hospitalar, sendo que 17 (7,9 %) episódios oc... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Community-acquired respiratory viruses (RV) are the most frequent etiologic agents causing acute respiratory infections (ARI) in humans. These agents have a wide antigenic range, universal distribution, affect people in all age groups, and may cause various clinical syndromes involving both the upper and lower respiratory tract. These respiratory infections are major causes of mortality in patients undergoing hematopoietic stem cell transplantation (HSCT), especially in the period prior to engraftment. These infections may also be acquired in hospitals, possibly transmitted by contact with infected health professionals or patient caregivers, or with contaminated objects or surfaces. Since 2008, a continued education program was started at the HSCT Program of Amaral Carvalho Foundation aiming to improve the control of RV transmission. Patients, caregivers, donors, family members and employees are invited to participate in the activities. Objectives: To review the cases of RV infections in patients undergoing HSCT from August 2010 to December 2013, characterize the type of transmission, if community- or hospital-acquired during this period, and determine the morbidity and mortality of RV infections. Methods: The study was conducted at the HSCT Service of Amaral Carvalho Hospital, analyzing the charts of HSCT recipients with RV infection diagnosed by immunofluorescent assay or multiplex PCR. Medical data and images from patients admitted to the HSC... (Complete abstract click electronic access below) / Mestre

Class of 2014 Abstract / Specific Aims: Busulfan is a chemotherapy used in conditioning regimens for hematopoeitic stem cell transplant (HSCT) that requires therapeutic drug monitoring (TDM) to reduce ther risk of adverse effects. Variable oral absorption and several studies demonstrating decreased toxicity with the intravenous formulation have led to IV preference despite the lower acquisition cost of oral busulfan. However, these studies failed to consider therapeutic drug monitoring and their results may therefore be flawed. The objective of this retrospective chart review was to determine the adverse effect, outcome profile, and cost-effectiveness of IV versus PO busulfan at a single medical center under TDM. Methods: This quality improvement project was a retrospective cohort analysis using patient data from a single large academic medical center from January 2007 to April 2013. Patients were included if they were 18 years or older and had undergone HSCT using either IV or PO busulfan using standard dosing regimens. This data was then used to design a cost-effectiveness model in order to determine if IV or PO busulfan is cost effective. Main Results: There were 68 subjects receiving autologous transplants and 37 subjects receiving allogeneic transplants that received busulfan as part of their pretreatment therapy and were included in this study. Allogeneic and autologous transplant populations were analyzed separately. In both populations there was no difference in occurrence of pulmonary toxicity, HVOD, or mucositis between the IV or PO groups. IV busulfan was significantly associated with an increased need for patient controlled analgesia in both autologous and allogeneic populations (p=0.038 and 0.028 respectively). Total cost of PO therapy was $30,081 and $30,047 less than IV for autologous and allogeneic transplants, respectively. PO therapy also represented a cost savings of $41 and $57 dollars for autologous and allogeneic transplants, respectively. This was confirmed through bootstrapping technique, which found PO to be dominant to IV busulfan. Conclusion: In conclusion, this study finds PO busulfan to be a therapeutically equivalent and cost saving option as part of a pretreatment regimen for both autologous and allogeneic hematopoietic stem cell transplants when therapeutic drug monitoring is performed.

Parvovirus B19 is a common human pathogen that typically infects erythroid progenitors and causes hematological problems such as anemia and aplastic crises. The clinical presentation depends mainly on the immunological status of the patient. PVB19 can cause serious clinical disorders in immunocompromised patients after transplantation. More than 1500 samples from 90 patients who passed the HSCT in 2015 were tested for the presence of PVB19 in this work. This work describes the incidence of the virus and two typical periods of onset of infection in patients after the transplantation. Although several sources report the negative effect of PVB19 infection on the survival of allogeneic graft patients, this work did not confirm this assertion. Also, the results of this work suggest that allogenic grafts are not the main source for transmission, but that it is likely to be reactivated after long-term persistent or latent PVB19 infections. PVB19 is divided into 3 genotypes. Genotype 1 is the most widespread, genotype 2 is very rare in Europe for the last 10 years, and genotype 3 occurs mainly in tropical localities. This work as the first describes the distribution of genotypes in the Czech Republic. More than 130 samples from 125 PVB19 positive patients, stored in the Motol University Hospital from 2004...

La maladie du greffon contre l’hôte (GvHD) est une complication majeure des greffes de cellules souches hématopoïétiques (HSCT) qui survient dans 30 à 70% des cas et peut causer la mort, malgré un traitement prophylactique bien conduit. Il existe donc une réelle demande clinique pour améliorer ces traitements prophylactiques. Parce que ces traitements prophylactiques reposent en général sur des agents immunosuppresseurs, ceux-ci contribuent à diminuer la reconstitution immunitaire du patient, ce qui a un impact défavorable sur les infections et les taux de rechute d’hémopathie maligne, et donc limite leur utilisation. Les immunoglobulines (IVIG) pourraient représenter une alternative intéressante puisqu’elles ont des propriétés immunomodulatrices et qu’elles sont de plus couramment utilisées en clinique pour traiter des patients ayant un déficit immunitaire. Leur capacité à réduire l’apparition et la sévérité de la GvHD, sans toutefois inhiber ou nuire à la reconstitution immunitaire chez le patient n’a néanmoins jamais été clairement démontrée. Les objectifs de ce projet sont donc d’évaluer l’efficacité des IVIG à réduire l’incidence et la sévérité de la GvHD dans un modèle murin humanisé de GvHD, ainsi que de déterminer le mécanisme d’action des IVIG. Ce modèle consiste à injecter des huPBMCs à des souris immunodéprimées ne pouvant les rejeter. Les résultats obtenus suggèrent que les IVIG possèdent un effet immunomodulateur permettant de réduire les signes cliniques et de retarder l’apparition de la GvHD, tout en permettant l’apparition de cellules NK. Les IVIG agiraient de façon indirecte sur les huPBMCs afin d’induire l’apparition des cellules NK. / Graft-versus-host disease (GvHD) is a major complication following hematopoietic stem cell transplantation (HSCT), occuring in 30 to 70% of HSCT despite proper prophylactic treatment, and can result in death. It is therefore primordial to improve the prophylactic treatments in order to reduce the frequency and severity of GvHD. Since these prophylactic treatments are based on the use of immunosuppressive agents, they inhibit the immune reconstitution and thus increase the risk of infections and relapse. Because of their immunomodulating properties, immunoglobulins (IVIG) represent a promissing alternative. Furthermore, IVIG are already widely used in patients with immune deficits. Since no study has yet clearly shown that IVIG can reduce GvHD without inhibiting the immune reconstitution in the patient, the scientific community is still debating their usefullness in GvHD prevention. Should their efficacy to reduce the incidence and severity of GvHD without impairment to the immune reconstitution be demonstrated, the use of IVIG could change the clinical outcome for HSCT patients, improving their remission and quality of life. The objectives of this project are to evaluate IVIG efficacy in GvHD prevention and to define the mechanism of action of IVIG. To fulfill these objectives, we will use a GvHD humanized mouse model, which consists of injecting huPBMCs in immunodeficient mice who can not reject them. Our results demonstrate an immunomodulatory effect of IVIG, allowing for reduced clinical signs and a slighlty increased survival in GvHD. A population of NK cells also appeared upon IVIG treatment and is believed to be indirectly induced by IVIG.

La maladie du greffon contre l’hôte (GvHD) est une complication majeure des greffes de cellules souches hématopoïétiques (HSCT) qui survient dans 30 à 70% des cas et peut causer la mort, malgré un traitement prophylactique bien conduit. Il existe donc une réelle demande clinique pour améliorer ces traitements prophylactiques. Parce que ces traitements prophylactiques reposent en général sur des agents immunosuppresseurs, ceux-ci contribuent à diminuer la reconstitution immunitaire du patient, ce qui a un impact défavorable sur les infections et les taux de rechute d’hémopathie maligne, et donc limite leur utilisation. Les immunoglobulines (IVIG) pourraient représenter une alternative intéressante puisqu’elles ont des propriétés immunomodulatrices et qu’elles sont de plus couramment utilisées en clinique pour traiter des patients ayant un déficit immunitaire. Leur capacité à réduire l’apparition et la sévérité de la GvHD, sans toutefois inhiber ou nuire à la reconstitution immunitaire chez le patient n’a néanmoins jamais été clairement démontrée. Les objectifs de ce projet sont donc d’évaluer l’efficacité des IVIG à réduire l’incidence et la sévérité de la GvHD dans un modèle murin humanisé de GvHD, ainsi que de déterminer le mécanisme d’action des IVIG. Ce modèle consiste à injecter des huPBMCs à des souris immunodéprimées ne pouvant les rejeter. Les résultats obtenus suggèrent que les IVIG possèdent un effet immunomodulateur permettant de réduire les signes cliniques et de retarder l’apparition de la GvHD, tout en permettant l’apparition de cellules NK. Les IVIG agiraient de façon indirecte sur les huPBMCs afin d’induire l’apparition des cellules NK. / Graft-versus-host disease (GvHD) is a major complication following hematopoietic stem cell transplantation (HSCT), occuring in 30 to 70% of HSCT despite proper prophylactic treatment, and can result in death. It is therefore primordial to improve the prophylactic treatments in order to reduce the frequency and severity of GvHD. Since these prophylactic treatments are based on the use of immunosuppressive agents, they inhibit the immune reconstitution and thus increase the risk of infections and relapse. Because of their immunomodulating properties, immunoglobulins (IVIG) represent a promissing alternative. Furthermore, IVIG are already widely used in patients with immune deficits. Since no study has yet clearly shown that IVIG can reduce GvHD without inhibiting the immune reconstitution in the patient, the scientific community is still debating their usefullness in GvHD prevention. Should their efficacy to reduce the incidence and severity of GvHD without impairment to the immune reconstitution be demonstrated, the use of IVIG could change the clinical outcome for HSCT patients, improving their remission and quality of life. The objectives of this project are to evaluate IVIG efficacy in GvHD prevention and to define the mechanism of action of IVIG. To fulfill these objectives, we will use a GvHD humanized mouse model, which consists of injecting huPBMCs in immunodeficient mice who can not reject them. Our results demonstrate an immunomodulatory effect of IVIG, allowing for reduced clinical signs and a slighlty increased survival in GvHD. A population of NK cells also appeared upon IVIG treatment and is believed to be indirectly induced by IVIG.