Made available in DSpace on 2014-06-11T19:27:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-03-04Bitstream added on 2014-06-13T20:16:32Z : No. of bitstreams: 1 lavezzo_lc_me_sjrp.pdf: 1608901 bytes, checksum: fe41b66f249b2bbe571fc89130581c47 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O Brasil é um país de dimensões continentais, com 8.512.000 km 2 , tem a maioria de seu território situado em área tropical, que associado a grande diversidade de flora e fauna, o que torna apropriada à ocorrência de zoonoses transmitidas por artropodes. Estudos sobre arbovírus na região, pelo Instituto Evandro Chagas, em 44 anos (1954-1998) identificaram um grande número de vírus, e pelo menos 187 diferentes arboviroses foram isoladas. A prevalência mundial de arboviroses cresceu dramaticamente nas últimas décadas, e nas áreas onde estas doenças são endêmicas, a transmissão por transfusão dificilmente é investigada. A preocupação com a transmissão de arboviroses, principalmente do Vírus Dengue (DENV), por transfusão sanguínea aumentou após a documentação da transmissão de West Nile Vírus (WNV) por transfusão nos EUA. DENV e WNV são semelhantes em alguns aspectos, o que suporta a hipótese de que dengue possa ser transmitida por transfusão sanguínea: são eficientemente transmitidos ao homem através da picada de mosquitos infectados; grande proporção das infecções é assintomática; níveis de viremia durante a fase de incubação pode exceder 106 virions por mL, e a transmissão de ambos foi documentada após transplante de órgão e em acidentes com profissionais da saúde. Desta forma, o objetivo deste trabalho foi investigar a presença de arboviroses em 205 amostras clínicas de doadores de sangue da Região Amazônica e em amostras obtidas durante uma epidemia de Dengue, em 2006, na cidade de Tupã/SP. O RNA viral foi extraído, Multiplex-Nested-PCR foi realizada com o uso de primers gênero e espécie-específico para Flavivirus, Alphavirus, e, para a detecção e identificação do vírus Oropouche, realizou-se RT- Nested-PCR usando primers que se ligam no segmento S do genoma viral. Apesar da complexidade e número, a transmissão... / O Brasil é um país de dimensões continentais, com 8.512.000 km 2 , tem a maioria de seu território situado em área tropical, que associado a grande diversidade de flora e fauna, o que torna apropriada à ocorrência de zoonoses transmitidas por artropodes. Estudos sobre arbovírus na região, pelo Instituto Evandro Chagas, em 44 anos (1954-1998) identificaram um grande número de vírus, e pelo menos 187 diferentes arboviroses foram isoladas. A prevalência mundial de arboviroses cresceu dramaticamente nas últimas décadas, e nas áreas onde estas doenças são endêmicas, a transmissão por transfusão dificilmente é investigada. A preocupação com a transmissão de arboviroses, principalmente do Vírus Dengue (DENV), por transfusão sanguínea aumentou após a documentação da transmissão de West Nile Vírus (WNV) por transfusão nos EUA. DENV e WNV são semelhantes em alguns aspectos, o que suporta a hipótese de que dengue possa ser transmitida por transfusão sanguínea: são eficientemente transmitidos ao homem através da picada de mosquitos infectados; grande proporção das infecções é assintomática; níveis de viremia durante a fase de incubação pode exceder 106 virions por mL, e a transmissão de ambos foi documentada após transplante de órgão e em acidentes com profissionais da saúde. Desta forma, o objetivo deste trabalho foi investigar a presença de arboviroses em 205 amostras clínicas de doadores de sangue da Região Amazônica e em amostras obtidas durante uma epidemia de Dengue, em 2006, na cidade de Tupã/SP. O RNA viral foi extraído, Multiplex-Nested-PCR foi realizada com o uso de primers gênero e espécie-específico para Flavivirus, Alphavirus, e, para a detecção e identificação do vírus Oropouche, realizou-se RT- Nested-PCR usando primers que se ligam no segmento S do genoma viral. Apesar da complexidade e número... (Complete abstract click electronic access below)

Orientador: Maurício Lacerda Nogueira / Banca: Luiz Carlos de Mattos / Banca: Maria Roberta Fachini / Resumo: O Brasil é um país de dimensões continentais, com 8.512.000 km 2 , tem a maioria de seu território situado em área tropical, que associado a grande diversidade de flora e fauna, o que torna apropriada à ocorrência de zoonoses transmitidas por artropodes. Estudos sobre arbovírus na região, pelo Instituto Evandro Chagas, em 44 anos (1954-1998) identificaram um grande número de vírus, e pelo menos 187 diferentes arboviroses foram isoladas. A prevalência mundial de arboviroses cresceu dramaticamente nas últimas décadas, e nas áreas onde estas doenças são endêmicas, a transmissão por transfusão dificilmente é investigada. A preocupação com a transmissão de arboviroses, principalmente do Vírus Dengue (DENV), por transfusão sanguínea aumentou após a documentação da transmissão de West Nile Vírus (WNV) por transfusão nos EUA. DENV e WNV são semelhantes em alguns aspectos, o que suporta a hipótese de que dengue possa ser transmitida por transfusão sanguínea: são eficientemente transmitidos ao homem através da picada de mosquitos infectados; grande proporção das infecções é assintomática; níveis de viremia durante a fase de incubação pode exceder 106 virions por mL, e a transmissão de ambos foi documentada após transplante de órgão e em acidentes com profissionais da saúde. Desta forma, o objetivo deste trabalho foi investigar a presença de arboviroses em 205 amostras clínicas de doadores de sangue da Região Amazônica e em amostras obtidas durante uma epidemia de Dengue, em 2006, na cidade de Tupã/SP. O RNA viral foi extraído, Multiplex-Nested-PCR foi realizada com o uso de primers gênero e espécie-específico para Flavivirus, Alphavirus, e, para a detecção e identificação do vírus Oropouche, realizou-se RT- Nested-PCR usando primers que se ligam no segmento S do genoma viral. Apesar da complexidade e número, a transmissão... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: O Brasil é um país de dimensões continentais, com 8.512.000 km 2 , tem a maioria de seu território situado em área tropical, que associado a grande diversidade de flora e fauna, o que torna apropriada à ocorrência de zoonoses transmitidas por artropodes. Estudos sobre arbovírus na região, pelo Instituto Evandro Chagas, em 44 anos (1954-1998) identificaram um grande número de vírus, e pelo menos 187 diferentes arboviroses foram isoladas. A prevalência mundial de arboviroses cresceu dramaticamente nas últimas décadas, e nas áreas onde estas doenças são endêmicas, a transmissão por transfusão dificilmente é investigada. A preocupação com a transmissão de arboviroses, principalmente do Vírus Dengue (DENV), por transfusão sanguínea aumentou após a documentação da transmissão de West Nile Vírus (WNV) por transfusão nos EUA. DENV e WNV são semelhantes em alguns aspectos, o que suporta a hipótese de que dengue possa ser transmitida por transfusão sanguínea: são eficientemente transmitidos ao homem através da picada de mosquitos infectados; grande proporção das infecções é assintomática; níveis de viremia durante a fase de incubação pode exceder 106 virions por mL, e a transmissão de ambos foi documentada após transplante de órgão e em acidentes com profissionais da saúde. Desta forma, o objetivo deste trabalho foi investigar a presença de arboviroses em 205 amostras clínicas de doadores de sangue da Região Amazônica e em amostras obtidas durante uma epidemia de Dengue, em 2006, na cidade de Tupã/SP. O RNA viral foi extraído, Multiplex-Nested-PCR foi realizada com o uso de primers gênero e espécie-específico para Flavivirus, Alphavirus, e, para a detecção e identificação do vírus Oropouche, realizou-se RT- Nested-PCR usando primers que se ligam no segmento S do genoma viral. Apesar da complexidade e número... (Complete abstract click electronic access below) / Mestre

Orientador: Cristina Antoniali Silva / Coorientador: Lusiane Maria Bendhack / Banca: Sandra Helena Penha de Oliveira / Banca: Virgínia Soares Lemos / Banca: Graziela Scalianti Ceravolo / Banca: Michele Paulo / Resumo: O uso clínico de drogas que liberam óxido nítrico (NO) é limitado por seus efeitos colaterais. A hipotensão induzida pelo doador clássico de NO, nitroprussiato de sódio (NPS) é rápida, transiente e induz à taquicardia reflexa, o que pode ser um efeito indesejável em pacientes com doença cardíaca e uma limitação para a terapia anti-hipertensiva. Este estudo avaliou o efeito hipotensor e vasodilatador do novo doador de NO [Ru(terpy)(bdq)NO+]3+ (TERPY) e comparou com os resultados obtidos com o NPS em ratos Wistar e ratos espontaneamente hipertensos (SHR). Em outra parte do estudo, foram estudadas diferenças no mecanismo de ação desta droga entre aortas de SHR jovens e velhos. Diferente do observado para o NPS, a hipotensão induzida pelo TERPY é lenta, duradoura e não leva a alterações da frequência cardíaca. Além disso, o TERPY libera quantidades semelhantes de NO em aortas de SHR e Wistar, induzindo relaxamento parcialmente dependente de GCs em ambos os grupos, ao contrário do NPS, que libera mais NO em aortas de SHR e também é mais potente e eficaz em aortas desses animais. Fatores como o estresse oxidativo e a atividade da PDE5 são importantes para o relaxamento do TERPY em SHR, mas a inibição da PDE5 não aumenta a potência do TERPY em aortas de ratos Wistar. Além disso, o relaxamento induzido pelo TERPY é mais potente em anéis de aorta de SHR velhos do que novos. Os mecanismos de ação do TERPY são semelhantes nas aortas desses animais, mas, interessantemente, a incubação com Apocinina aumenta a potência do TERPY em aortas de SHR jovens, mas não de velhos. Em conjunto, estes dados demonstram que o composto TERPY é um doador de NO que possui vantagens em relação ao NPS. Além disso, é mais potente em aortas de animais hipertensos velhos, o que é mais uma vantagem para sua utilização e um incentivo para a realização de novos estudos que possam contribuir para entender melhor seu mecanismo de ação / Abstract: The clinical use of nitric oxide (NO) releasing drugs is limited by their harmful effects. The hypotension induced by the classic NO donor, sodium nitroprusside (SNP) is fast, transient and induces reflex tachycardia, which can be an undesirable effect in patients with heart disease and a limitation for the anti-hypertensive therapy. This study evaluated the hypotensive and vasodilatory effects of the new NO donor [Ru(terpy)(bdq)NO+]3+ (TERPY) in Wistar rats and spontaneously hypertensive rats (SHR). In another part of the study, we investigated the differences in the mechanism of action of this drug between aortas of young and old SHR. Different from what is observed for SNP, the hypotension induced by TERPY is slow, long lasting and doesn't lead to alterations in the heart rate. Besides, TERPY releases similar amounts of NO in SHR and Wistar aortas, inducing a relaxation partially dependent on GCs in both groups, contrary to SNP, which releases more NO in aortas of SHR and is also more potent and efficient in the aortas of these animals. Factors as oxidative stress and the activity of PDE5 are important to the relaxation of TERPY in SHR, but the inhibition of PDE5 doesn't increase the potency of TERPY in aortas of Wistar rats. Furthermore, the relaxation induced by TERPY is more potent in aortas of old SHR than of young ones. The mechanisms of action of TERPY are similar in the aortas of both groups, but, interestingly, the incubation with Apocynin increases the potency of TERPY in aortas of young SHR, but not of old ones. Taken together, these data show that the compound TERPY is a NO donor that has advantages in relation to SNP. Moreover, it's more potent in aortas of old hypertensive animals, which is another advantage for its use and an incentive for the elaboration of new studies that could contribute to understand its mechanism of action / Doutor

The one-way mixed lymphocyte culture (MLC) is used to assess histocompatibility between donor and recipient. First introduced in 1966, this method involves the co-culture of lymphocytes from the peripheral blood of the donor and the recipient for a period of 6 to 7 days: antigen disparities, primarily in the HLA-DR region, stimulate proliferation of the responding cells, which is detected by addition of 3H-labelled thymidine and subsequent measurement of radioactivity. The lymphocytes of either the donor, used to predict graft-versus-host disease (GVHD) or recipient, used to predict host-versus-graft disease (HVGD)/graft rejection, are inactivated by exposure to radiation or mitomycin C, so that the observed proliferation is that of the other set of lymphocytes, hence the name “one-way” MLC. The amount of measured radioactivity is directly proportional to the amount of DNA synthesized, which is a reflection of the number of disparities at the major histocompatibility complex (MHC).Previous studies have established that inactivation of the lymphocytes by radiation and mitomycin C, has a negative effect on the structure/expression of HLA-DR molecules on the cell surface, which provides the primary stimulus for the MLC reaction. The laboratory research presented in this dissertation was designed i) to compare the viabilities and HLA-DR levels on stimulator cells exposed to cyclosporin A, mitomycin C and ionizing irradiation , in order to determine whether cyclosporin A can be used as an alternative to mitomycin C or radiation as inactivator of the stimulator cells in the one-way MLC; ii) to improve sensitivity and accelerate the MLC reaction by addition of IL-2; iii) establish a flow cytometric mixed lymphocyte assay using the fluorochrome 5,6 carboxyfluorescein diacetate succinimidyl ester (CFSE). Cyclosporin A showed striking similarities to mitomycin C and ionizing radiation in its effect on viability and reduction/structural changes in HLA-DR molecules of the stimulator cells. Exposure of the stimulator cells to 20ìM cyclosporin A, demonstrated a significant loss of both cell viability and HLA-DR molecule cell surface expression. Thus, in evaluating these three methods of inactivation of the stimulator cells in the one-way MLC, it was concluded that a one-way MLC may not in fact be an accurate and qualitative reflection of the histocompatibility between donor and recipient. Instead the two-way MLC , in which neither the donor’s nor recipient’s cells are inactivated, may be a more reliable alternative. The only limitation associated with a two-way MLC is the inability to distinguish between a host-versus-graft-rejection and a graft-versus-host reaction in the observed allogeneic response Addition of 5 and 10 IU/ml IL-2 to the MLC showed the opposite effect to that intended, inhibiting proliferation in the MLC. Previous studies have shown that an excess of IL-2 results in the production of suppressor T cells. The amount of IL-2 produced during the MLC depends on the number of disparities in the MLC between donor and recipient, which will be different for each MLC reaction. Since the number of allogeneic T cells involved in the MLC reaction is not known, the amount of IL-2 produced during the allogeneic immune response in the MLC can not be predicted and addition of exogenous IL-2 may result in production of suppressor T cells and an inhibition of proliferation. The two-way MLC was modified by staining one of the participating set of lymphocytes (donors or recipients) with CFSE and tracking proliferation in this population, using flow cytometry. The two-way CFSE-based MLC analyzed in this study were counterstained with CD25 (IL-2R). An increase in CD25 expression on the cell surface is an indicator of cell activation and proliferation. Proliferation, as indicated by a progressive loss of CFSE fluorescence correlated well with the corresponding increase in CD25 expression and accumulated daughter cells. In addition, by loading only one of the participating donors in the two-way MLC, the responder/stimulator interaction, observed in the one-way MLC, is re-established. Thus the modified, CFSE-based two-way MLC can be used to predict GVHD. To conclude, the use of CFSE labeling and flow-cytometry to measure proliferation in a two-way MLC, together with CD25 counterstaining provides an alternative, reliable and probably superior method to 3H thymidine uptake. / Dissertation (MSc)--University of Pretoria, 2009. / Immunology / unrestricted

Maternal consumption of dietary methyl donors (DMDs) such as folate, methionine, choline, as well as co-factors including zinc, vitamins B2, B6 and B12 can lead to permanent alterations in the DNA and gene expression of the developing fetus. This study aimed to identify patterns of DMD intake during the second trimester of pregnancy and their associations with infant birth weight, small and large for gestational age (SGA and LGA, respectively). From food sources alone, most pregnant women were below the estimated average requirement for dietary folate equivalent (DFE) (69%) and below the adequate intake measure for choline (99%). Zinc seemed to be the most important nutrient for attaining adequate birth weight. DFE and vitamin B12 were positively associated with birth weight. DFE also reduced the risk for SGA whereas choline increased the risk for LGA. Therefore, DMD intake from food sources during pregnancy may be important to ensure optimal infant birth weight.

The main aim of this thesis is to reveal the importance of the development aid in Kenya and to demonstrate, on the example of Slovak republic, which type of aid plays more important role -- wheather the official or unofficial aid. Effectivness of aid in Kenya is evaluated on the basis of correlation of the amounts of development aid and indices which show improvement or degradation of economic and social situation of the country. The significance of the official and unofficial aid is evaluated by comparing the projects that are supported by ODA and projects supported by non-governmental organizations on the basis of financial criterium and the reach of the projects. The contribution of this thesis is to show effectivness of development aid in poor countries on the example of Kenya. The work has three chapters. The object of the first chapter is to present economic and social situation in Kenya and individual factors which can help her to get over the underdevelopment. The second chapter offers characteristics of individual donors and evaluates the relevance of development aid. The last chapter presents concrete projects of ODA of Slovak republic and of Slovak non-governmental organizations and evaluates which of the two types of aid is more important.

Nonprofit organization senior leaders conduct business in an increasingly complex and competitive market environment relying on monetary donations to sustain the capacity of their organizations to meet service requirements. The purpose of this single case study was to explore the strategies that nonprofit organization senior leaders used to motivate donors to make monetary donations to their organizations. The target population of the study was 3 senior leaders of a nonprofit organization in the southeastern United States who implemented effective strategies to motivate donors to make monetary donations to their organization. The conceptual framework used for this study was self-determination theory. Data collection for this study consisted of semistructured participant interviews, a review of public and internal organization documentation, and organization performance outcomes. Data for this study were manually coded and thematically organized. The analysis of the data identified relationships, effective communication, active listening, and social media marketing as 4 key themes affecting donors’ motivation to donate to nonprofit organizations. The implications of this study for positive social change include the potential to provide practical strategies that nonprofit organization senior leaders can use to motivate donors to make monetary donations to their organizations, thereby enhancing senior leaders’ capacity to provide critical services to members of the communities in their organizations’ dedicated service areas.

Oligodendrocytes are predominately found in white matter of the brain, but also populate gray matter regions. Although commonly known to provide myelination of neuronal axons, these cells serve numerous other functions in the brain. A unique property of oligodendrocytes is their inherent susceptibility to oxidative stress because of several biochemical characteristics of these cells, including a high concentration of iron, high metabolic rate, and low antioxidant enzyme activity. Oxidative stress conditions are produced by inflammation, and both inflammation and oxidative stress are highly associated with major depressive disorder (MDD). Hence, the study of oligodendrocytes in the brain in MDD readily provides access to molecular mechanisms engaged by oxidative stress conditions that putatively contribute to the etiology of MDD. My laboratory studied oligodendrocytes, and other white matter cells, from postmortem tissue collected from brain donors that died as a result of suicide and other causes, focusing on those donors who had at the time of death either MDD or no psychiatric or neurologic diagnosis (controls). White matter oligodendrocytes or whole white matter in limbic brain from MDD/suicide donors demonstrated indices of elevated oxidative damage, including increased DNA oxidation, shortened telomere DNA, reduced expression of antioxidant enzyme genes, and upregulated DNA base excision repair enzymes. These abnormalities were either not observed or were only modestly evident in astrocytes collected from white matter of the same MDD/suicide donors. To determine whether this oxidative damage was restricted to white matter in the limbic brain, oligodendrocytes were captured from three other brain regions, prefrontal cortical (BA 10) white matter, occipital cortical white matter, and gray matter in the region of the brainstem locus coeruleus. Shortened telomeres and reduced expression of antioxidant enzyme genes were observed in oligodendrocytes from these additional brain regions in MDD/suicide. Since this oligodendrocyte pathology was not anatomically restricted to the limbic brain, it may be difficult to understand how it is relevant to the biological basis of emotional behaviors that are specifically associated with MDD or suicide. However, the oligodendrocyte is highly susceptible to oxidative stress; hence, the oligodendrocyte can be viewed as a “canary in the coal mine” for detecting oxidative damage to the brain. Therefore, elucidation of the molecular pathways activated by oxidative damage in these cells could reveal novel targets for the development of drugs to prevent oxidative damage and its subsequent pathological activation of downstream pathways deleterious to brain cell health. As such, drugs targeting these pathways may have antidepressant properties in humans, and could provide an alternative approach to treating depression and reducing suicide risk. In fact, we found that repeated exposure of rats to psychological stress increased DNA oxidation in prefrontal cortical white matter. Furthermore, preliminary findings using rat models of depression reveal that interruption of pathways downstream to oxidative damage produces a robust antidepressant response, correcting depressive-like behaviors elicited by psychological stress. These findings strongly implicate a role of oxidative damage in the etiology of MDD and possibly suicide, and demonstrate the utility of studying brain pathology as a logical path to identifying novel antidepressant targets.

Nonprofit agencies are instrumental in U.S. society and local communities. While there is significant outreach targeted for wealthy and middle-class Caucasian families, there is a lack of targeted effort for African Americans. The purpose of this case study was to explore the strategies development directors use to engage and retain African American donors. The focus of the research questions was what factors lead to successful engagement campaigns for African Americans. The qualitative case study was suitable for gathering data from multiple sources of information, including an interview questionnaire and existing literature. A purposeful sampling strategy was appropriate for choosing 5 development directors from 5 nonprofit organizations for the open-ended interviews. Transcribed data were analyzed by comparing responses that led to theme creation. Source triangulation utilized for trustworthiness and response interpretation revealed community development, fostering relationships and intentional engagement are themes that attract African Americans. Results of the study showed development directors who utilize community based and family oriented themes and targeted social media campaigns reach more African American patrons. The social implications outlined in this study are intended to assist nonprofit leaders create a stable donor base so they can positively impact the local community. By addressing engagement strategies with a significant portion of the community, nonprofit leaders can attract and retain this largely untapped market for continued sustainability.

This study measures two reinforcement strategies designed to increase repeat donations in first-time volunteer blood donors.