Tese (doutorado)—Universidade de Brasília, Faculdade de Medicina, 2008. / Submitted by Kelly Marques (pereira.kelly@gmail.com) on 2009-10-21T18:46:59Z No. of bitstreams: 1 2008_AlessandraMenezesCampos.pdf: 1238572 bytes, checksum: dab07da7d1c9342e2084f202e412302b (MD5) / Approved for entry into archive by Tania Milca Carvalho Malheiros(tania@bce.unb.br) on 2009-10-30T12:18:22Z (GMT) No. of bitstreams: 1 2008_AlessandraMenezesCampos.pdf: 1238572 bytes, checksum: dab07da7d1c9342e2084f202e412302b (MD5) / Made available in DSpace on 2009-10-30T12:18:22Z (GMT). No. of bitstreams: 1 2008_AlessandraMenezesCampos.pdf: 1238572 bytes, checksum: dab07da7d1c9342e2084f202e412302b (MD5) Previous issue date: 2008 / A renina é uma aspartil protease sintetizada in vivo pela remoção proteolítica do prósegmento amino-terminal de seu precursor, a pró-renina. A catepsina B, uma tiol protease, tem sido sugerida como candidata à enzima conversora de pró-renina devido à sua colocalização com a renina nos grânulos secretórios e também pela sua capacidade de processar a pró-renina em renina in vitro e em cultura de células. Vetores de expressão de pró-renina selvagem (WT) e mutantes foram co-transfectados com catepsina B em células hipofisárias de ratos GH4C1, que naturalmente não converte pró-renina em renina. Mutações pontuais foram utilizadas para identificar quais aminoácidos presentes no sítio de clivagem da pró-renina são reconhecidos pela catepsina B. Experimentos de pulse-chase foram realizados para rastrear se a pró-renina radiomarcada é capaz de seguir para os grânulos secretórios das células GH4C1. Para investigar se a pró-renina humana poderia ativar o promotor do Peptídeo Natriurético Cerebral (BNP) em cardiomiócitos de ratos neonatos, essas células foram co-transfectadas com o plasmídeo repórter BNP-luciferase com (ou sem) angiotensinogênio humano. Os cardiomiócitos foram co-incubados com as células não-aderentes U937 transfectadas com pró-renina humana WT ou mutantes (-1-2 ou +5). Os cardiomiócitos foram lisados 96 horas após a eletroporação para a análise de geração de luciferase. Células não-transfectadas U937 foram usadas como grupo controle. Captopril e Losartan (10-7 M) foram usados para testar a participação da enzima conversora de angiotensina (ECA) e dos receptores AT1. Os resultados demonstraram que mutações pontuais na metionina -3(M/A), lisina -2(K/A) e leucina +2(L/A) reduzem o processamento da pró-renina pela catepsina B. Além disso, a remoção da cadeia glicosídica na asparagina +5 aumenta a conversão de pró-renina, sugerindo que a glicosilação poderia impedir o processamento enzimático da pró-renina. Nos experimentos de pulse-chase, a pró-renina não foi direcionada aos grânulos secretórios e a presença de catepsina B não alterou esse resultado, indicando que o processamento de pró-renina provavelmente ocorre fora dessas organelas. A pró-renina WT secretada pelas células U937 é capaz de ativar o promotor do BNP em cardiomiócitos de ratos transfectados com angiotensinogênio humano. O par de aminoácidos básicos arginina-lisina e a cadeia glicosídica +5 parecem ser tão importantes quanto a atividade enzimática da ECA e os receptores AT1 para a ativação do promotor do BNP em cardiomiócitos de ratos neonatos. ____________________________________________________________________________________ ABSTRACT / Renin is an aspartyl protease synthesized in vivo by proteolytic removal of amino-terminal prosegment of its precursor, prorenin. Cathepsin B, a thiol protease, has been proposed to be a good candidate for prorenin processing enzyme because of its co-localization with renin in secretory granules and also by its ability to activate prorenin in vitro and in cell culture. It was co-transfected vectors expressing wild type and mutant prorenins with cathepsin B in rat pituitary GH4C1 cells, which normally do not activate prorenin into renin unless cotransfected with a prorenin processing enzyme. Scanning mutagenesis was used to identify which amino acids at prorenin cleavage site are specifically recognized by cathepsin B. Pulsechase experiments were performed to follow whether radiolabeled prorenin is sorted to secretory granules. To investigate whether human prorenin could activate the Brain Natriuretic Peptide promoter (BNP) in neonatal rat cardiomyocytes, these cells were cotransfected with a reporter gene BNP-luciferase associated or not with a human angiotensinogen. The cardiomyocytes were co-incubated with no adherent U937 cells transfected with human prorenin wild type (WT) or mutants (-1-2 or +5). Cardiomyocytes were lysated 96 hours after electroporation for luciferase generation analysis. No transfected U937 cells were used as a control group. Captopril and Losartan at 10-7 M were used to test angiotensin converting enzyme (ACE) and AT1 receptors participation in this process. The results demonstrated that single mutations at - 3 methionine (M/A), -2 lysine (K/A) and +3 leucine (L/A) reduced prorenin processing by cathepsin B. Moreover, removal of N-linked oligosaccharides at +5 asparagine enhanced prorenin conversion by cathepsin B, showing that glycosylation seems impair enzymatic prorenin processing. In pulse-chase experiments, prorenin was not targeted to secretory granules and cathepsin B co-transfection did not changed this sorting, indicating that prorenin processing probably occurs outside these organelles. These findings suggest that cathepsin B removes prorenin prosegment at correct cleavage site and this proteolysis occurs outside the dense core granules. Prorenin WT secreted by U937 cells could activate the BNP promoter in rat cardiomyocytes transfected with human angiotensinogen. Dibasic arginine-lisine prorenin amino acids and N-linked glycosilation seems to be important as well as ACE enzymatic activity and AT1 receptors for BNP promoter activation.

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A hepatite B é uma doença viral infectocontagiosa mundialmente disseminada. É caracterizada por ser uma inflamação hepática que possui risco de cronicidade em 10 a 20% dos casos ocorridos em adultos. Atualmente, acredita-se que aproximadamente 240 milhões de pessoas estejam cronicamente infectadas. A prevalência da doença varia em todo o mundo. Embora o Brasil seja considerado um país de endemicidade baixa, existem regiões onde a prevalência da doença é acentuadamente superior à média nacional, como a região Amazônica, e a região oeste catarinense. A imunopatogênese da infecção por sua vez, é influenciada tanto por fatores relacionados ao vírus da hepatite B (HBV), como genótipo, carga viral e mutações genéticas, quanto a características do hospedeiro infectado, como idade, sexo e polimorfismos genéticos. O presente trabalho buscou determinar fatores virais e do hospedeiro que estejam influenciando no curso natural da hepatite B, através da análise do perfil molecular viral e de polimorfismos de base única (SNPs) de genes de citocinas de pacientes infectados, estabelecendo possíveis marcadores moleculares de progressão da doença. Neste estudo, foram coletadas 363 amostras de indivíduos infectados com HBV em Chapecó, SC. Em relação aos fatores virais analisados, encontramos uma baixa proporção de indivíduos (15,3%, 31/202) com carga viral acima de 2000UI/mL e uma frequência de 5,6% (9/161) de hepatite oculta. A genotipagem viral foi realizada em 91 amostras e destas, 88 (97%) apresentaram HBV/D e 3 (3%) HBV/A, contrastando com a realidade de outras regiões brasileira, onde há um predomínio de HBV/A (64,3-44,7%), seguidos de HBV/D (47,6-16,6%) e HBV/F (29,2-7,7%). A análise de subtipos virais revelou uma alta frequência do subtipo ayw2. Para justificar a frequência elevada de HBV/D subtipo ayw 2 na região sul, foi realizada uma busca pela origem genealógica e geográfica dos pacientes infectados e uma análise filogenética foi executada. Os resultados mostraram que o HBV/D encontrado no sul do Brasil é filogeneticamente muito próximo ao HBV/D3 amplamente disseminado na Itália, assim como para o subtipo ayw2, e sua alta frequência teria poderia ser justificada pelo processo de imigração italiana ocorrido durante o final no século XIX e início do século XX. Entre os fatores do hospedeiro, foi realizada uma análise comparativa entre pacientes cronicamente infectados e com infecção resolvida, avaliando variáveis epidemiológicas e genéticas. A análise epidemiológica mostrou que sexo masculino, via de transmissão sexual e baixo nível educacional estão associados a cronicidade da doença. Os genótipos dos SNPs foram analisados através de regressão logística e os resultados mostraram que o alelo A (genótipos AA e GA) do SNP -308 do fator de necrose tumoral-α (TNF-α) está associado à cronicidade da hepatite B em indivíduos do sul do Brasil e pode ser considerado um potencial marcador molecular de progressão à cronicidade da hepatite B nesta população. / Hepatitis B is a viral infectious disease spread worldwide. It is a liver inflammation with a chronicity risk about 10 to 20% of adulthood cases. Currently, approximately 240 million people are chronically infected. The disease prevalence varies worldwide. Although Brazil is considered a low endemic country, there are regions where the prevalence is markedly higher than the national average, such as the Amazon region (Northern Brazil) and the west region of Santa Catarina, in southern Brazil. The immunopathogenesis of infection is influenced by factors relating to hepatitis B virus (HBV), such as genotype, gene mutations and viral load, and by characteristics of the infected host, such as age, sex and genetic polymorphisms. This study aimed to determine viral and host factors that are influencing on the natural course of the hepatitis B, through the viral molecular profile analysis and single nucleotide polymorphisms (SNPs) genotyping of cytokines genes from patients infected in an attempt to suggest molecular markers of disease progression. In this study, were collected 363 samples from individuals infected with HBV in Chapecó, SC. Regarding viral factors analyzed, we found a low proportion of individuals (15.3%, 31/202) with viral load above 2000 IU/mL and a frequency of 5.6% (9/161) of occult hepatitis infection (OBI). Viral genotyping was performed on 91 samples. Of these, 88 (97%) had HBV/D and three (3%) had HBV/A, in contrast with the reality of other Brazilian regions where there is a predominance of HBV/A (64.3 to 44.7%), followed by HBV/D (47.6 to 16.6%) and HBV/F (29.2 to 7.7%). The viral subtypes analysis has revealed a high frequency of subtype ayw2. To justify the high frequency of HBV/D and subtype ayw2 in the Brazilian southern region, a search for genealogical and geographical origin of the patients was performed and a phylogenetic analysis was conducted. The results showed that HBV/D in southern Brazil is phylogenetically close to HBV/D3 widespread in Italy as well as the ayw2 subtype and its high frequency would could be justified by the Italian immigration process occurred during late nineteenth and early twentieth century. Among the host factors, were conducted a comparative analysis of chronically infected patients and HBV resolved subjects, evaluating epidemiological and genetic variables. Epidemiological analysis showed that male sex, vertical transmission and low educational level have an association with chronic disease. The genotypes of the SNPs were analyzed by logistic regression and the results showed that the A allele (AA and GA genotypes) of tumor necrosis factor-α (TNF-α) -308 SNP is associated with chronic hepatitis B in individuals of southern Brazil and can be considered a potential molecular marker of progression to chronic hepatitis B in this population.

Os fatores que afetam a susceptibilidade a infecção com E. granulosus são pouco conhecidos. Neste trabalho, foi avaliada a interação de protoescólices e um antígeno imunodominante secretado (AgB) de E. granulosus com neutrófilos humanos saudáveis. A intensidade de expressão de CD11b e produção de H2O2 foram avaliadas por citometria de fluxo usando sangue total. Os neutrófilos isolados também foram incubados com protoescólices ou AgB, e a interleucina 8 (IL-8) foi determinada por ELISA de captura. Os protoescólices induziram um aumento significativo na expressão de CD11b em neutrófilos (p=0,026) e na produção de H2O2 (p=0,026), entretanto a sua viabilidade não foi afetada. Em contraste, nenhum efeito foi observado com o AgB em ambos ensaios. Uma moderada produção de IL-8 foi detectada somente em sobrenadantes de neutrófilos cultivados com protoescólices comparada com a produção basal (p=0,028). O efeito de uma incubação prévia com o AgB foi avaliada em relação a expressão de CD11 e H2O2 induzida pelo tratamento dos neutrófilos com forbol meristato acetato (PMA). Nenhuma mudança foi observada na expressão de CD11b, entretanto, a intensidade de produção de H2O2 foi significantemente reduzida em comparação ao tratamento apenas com PMA (p=0,038) quando o fator ativador de plaquetas (PAF) foi usado como priming. Como os protoescólices ocasionalmente podem ser liberados dos cistos em compartimentos do hospedeiro causando a hidatidose secundária, estes resultados contribuem para elucidação da defesa do hospedeiro e dos mecanismos imunomoduladores do parasito. A identificação de produtos de secreção e/ou excreção de protoescólices de E. granulosus também foi avaliada neste trabalho. Os produtos de secreção e/ou excreção foram obtidos a partir das primeiras 48 h de cultivo de protoescólices in vitro. A caracterização imunológica foi feita por western blotting com um pool de soros de indivíduos infectados com hidatidose cística. Os produtos de secreção e/ou excreção também foram analisados por espectrometria de massas. As preparações de seis diferentes amostras de protoescólices continham componentes protéicos, os quais foram visualizados por SDS-PAGE com aparentes massas moleculares entre 10 e 200 kDa. Os ensaios de western blotting mostraram a presença de diversos antígenos imunogênicos de massas moleculares maiores (acima de 50 kDa), e do AgB. Os resultados de espectrometria de massas foram obtidos com um pool de seis amostras de sobrenadantes de cultivo contendo produtos de secreção e/ou excreção, e foram identificadas onze proteínas: malato desidrogenase, ciclofilina, miofilina, P-29, 14-3-3, enolase, duas serinoproteases, fosfoenolpiruvato carboxiquinase e tiorredoxina peroxidase. Estes resultados também podem contribuir para elucidação de novos aspectos da complexa interação parasito-hospedeiro, e de mecanismos imunomoduladores, uma vez que as proteínas secretadas podem estar diretamente envolvidas nesses processos. / The factors affecting innate susceptibility to infection with E. granulosus are little known. In this work, the interaction of healthy human neutrophils with protoscoleces and a secreted immunodominant antigen (AgB) of E. granulosus was assessed. The intensity of CD11b expression and H2O2 production were evaluated by flow cytometry using whole blood. Isolated neutrophils were incubated also with PSC or AgB and interleukin 8 (IL-8) was measured by capture ELISA. The protoscoleces induced a significant increment of CD11b expression in neutrophils (p=0.026) and of H2O2 production (p=0.026), however their viability was not affected. In contrast, no effect was observed with AgB in both assays. A moderate IL-8 production was detected only in supernatants from neutrophils cultured with protoscoleces compared with basal production (p=0.028). The effect of previous AgB incubation was evaluated in relation with CD11b and H2O2 expression induced by phorbol meristate acetate (PMA) treatment. No changes were observed with CD11 expression, however the magnitude of H2O2 production was significant decreased in comparison with PMA alone (p=0.038) when platelet–activating factor was used for priming. As protoscoleces occasionally spillaged from cysts into compartments of the host causing secondary cysts, these results contribute to the elucidation of the host defense and parasite immunomodulatory mechanisms involved. The identification of excretory and/or secretory products from E. granulosus protoscoleces also was assessed in this work. Excretory and/or secretory products were obtained from the first 48 h maintenance of protoscoleces in vitro. Immunological characterization was performed by western blotting with a pool of sera from individuals infected by cystic hydatidosis. Excretory and/or secretory products also were assessed by mass spectrometry. The preparations of six differents sample of protoscoleces contained components proteins which could be distinguished by SDS-PAGE with apparent molecular masses between 10 and 200 kDa. The assays of western blotting showed the presence of the several immunogenic antigens of major molecular masses (above 50 kDa), and of the AgB. The results of the mass espectrometry were obtained with a pool of six cultive sobrenadants sample containing excretory and/or secretory products, and were identified eleven proteins: malate dehydrogenase, cyclofilin, myophilin, P-29, 14-3-3, enolase, tegumental protein, two serineproteases, phosphoenolpyruvate carboxykinase and thioredoxin peroxidase. These results can contribute also for elucidate novel aspects of the complex interaction host-parasite and of the immunomodulatory mechanisms, since that secreted proteins can be directly involved in these process.

Orientador: Fernando Lopes Gonçalves Junior / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-07-22T00:38:43Z (GMT). No. of bitstreams: 1 Boccato_RaquelSilveiraBelloStucchi_D.pdf: 7892385 bytes, checksum: 46ca59a347caf357ef35630271f4b26a (MD5) Previous issue date: 1996 / Resumo: Os profissionais da área da saúde (P AS) apresentam risco de infecção pelo VHB três a cinco vezes mais alto que a população geral. Entretanto, o alto custo da vacina é um empecilho para que se possa garantir uma ampla cobertura vacinal. Vários estudos têm sido realizados no sentido de tentar-se viabilizar esquemas vacinais mais econômicos, mantendo a eficácia. A aplicação intradérmica reduz de sete a dez vezes o custo da vacinação e, em determinados grupos populacionais, apresenta eficácia semelhante à da intramuscular. Tentando buscar esquemas alternativos mais econômicos, para vacinação contra o VHB em P AS, selecionamos um grupo de funcionários da área da saúde da Unicamp para receberem a vacina contra o VHB. Estes profissionais foram divididos em dois grupos, conforme a via de aplicação: intradérmica (ID) ou intramuscular (1M). Todos os que foram alocados para aplicação ID e uma parcela dos que a receberam via 1M foram acompanhados para avaliação da resposta imunológica à vacinação, através da pesquisa qualitativa e quantitativa do anti-HBs. O esquema proposto para a vacinação foi de três doses, aplicadas nos intervalos de um e seis meses, em relação à primeira dose. A vacina utilizada foi a vacina recombinante belga ("Engerix B", SmithKline), na dose de 20 µ g (1 ml) para aplicação 1M e 0,2 µ g (0,1 ml) para aplicação ID (face ventral do antebraço esquerdo). De todos os funcionários que receberam a primeira dose da vacina fez-se a coleta de sangue, para determinação da prevalência do anti-HBc, como niarcador da infecção pregressa pelo VHB. Novas amostras foram colhidas no momento da aplicação das segundas e terceiras doses e seis meses após esta última para a pesquisa do anti HBs. Os profissionais que completaram todo este acompanhamento e permaneceram na instituição foram convocados, depois de cinco anos, para nova titulação do anti-HBs e pesquisa do anti-HBc. Nesta ocasião, aos que exibiram títulos de anti-HBs < 10 mUI/ml, após terem apresentado soro conversão com o esquema primário, foi administrada uma dose de reforço da vacina (20 µ g), via 1M. Após duas a quatro semanas, nova titulação do anti-HBs foi realizada. Compareceram para a primeira dose da vacina 1117 funcionários, sendo que 463 (41,5%) receberam a vacina via ID e, entre os 654 (58,5%) da via 1M, 228 foram alocados para constituírem o subgrupo de acompanhamento da soroconversão. A prevalência do anti-HBc entre os 1117 P AS foi de 10,3%. Determinadas categorias profissionais, ou especialidades, apresentaram prevalências mais altas, como laboratoristas anestesistas, anatomopatologistas e radiologistas. Observamos também maiores prevalências do anti-HBc nos indivíduos com idade superior a 35 anos e naqueles que atuam na área da saúde há mais de dez anos. Entre os 115 PAS reagentes para o anti-HBc, dois deles apresentaram HBsAg reagente e os demais eram positivos para o anti-HBs, revelando uma prevalência de portadores do VHB de 0,2%. A adesão entre a primeira e segunda doses foi de 81,8% e entre a segunda e terceira doses de 79,4%. No grupo intradérmico, entretanto, o absenteísmo foi de 29,3%. Em 77,4% dos vacinados contra o VHB, a aplicação foi segura e bem tolerada, não causando reações adversas. As reações foram mais freqüentes no grupo ID que no grupo 1M (31,4 e 15,7%; p=0,000). No grupo ID, predominaram as reações locais (hiperemia, prurido e edema no local de aplicação) e no grupo 1M, as reações sistêmicas (sonolência, mialgia, náusea)~ Houve um caso de reação anafilática após a aplicação via intradérmica da segunda dose da vacina. Após três doses da vacina, a soroconversão para o anti-HBs foi de 92,2% no grupo 1M e de 59,9% no grupo ID. No grupo 1M, obtivemos títulos de anti-HBs < 10 mUI/ml em 24,5%, títulos entre 10 e 100 mUI/ml em 13,7% e títulos acima de 100 mUI/ml em 61,8%. No grupo ID, estes títulos foram observados em 46,4%, 34,3% e 19,3%, respectivamente. Idade inferior a trinta anos e pertencer ao sexo feminino foram fatores associados a uma adequada resposta vacinaI. Após cinco anos do esquema vacinal, observamos títulos de anti-HBs >= 10 mUI/ml em 73,7 e 43,2% dos servidores dos grupos 1M e ID, respectivamente. Ao administrarmos uma dose de reforço em 36 funcionários com anti-HBs < 10 mUI/ml, 88,9% apresentaram títulos de anti-HBs protetores e 83,3% exibiram títulos superiores a 150 fi UI/mI / Abstract: Health care workers (HCW) present HBV risk of infection three to five times higher than the population in general. However, high vaccine cost is an obstacle to assure fulI vaccinal coverage. Several studies have been made in order to try to make the vaccinal scheme more economical but effective. Intradermic HBV vaccine reduces seven to ten times the cost of vaccination and in certain population groups its effectiveness is similar to intramuscular application. In order to find altemate and more economic schemes for vaccination against HBV in HCW, we have selected a group of health care workers at Unicamp to receive the HBV vaccine. These workers have been divided into two groups, according to the application route: intradermic (ID) or intramuscular (1M). All the workers who received ID application and part of those who received the vaccine via 1M were followed up for evaluation of the immunologic response to the vaccine through both quantitative and qualitative anti-HBs investigation. The proposed vaccination scheme was of three doses, at O, 1 and 6 months intervals. The vaccine used was the Belgium recombinant vaccine (Engerix B, SmithKline), with 20 µ g (1 ml) dose for 1M application and 0.2 µ g (0.1 ml) dose for ID application (ventral side of the left forearm). AlI the workers receiving the first vaccine dose were submitted to a blood test to determine the prevalence of anti-HBc as a marker of the previous HBV infection. New blood samples were collected at the time the 2nd and 3rd doses were applied and 6 months after the last one for anti-HBs survey. The workers who completed the program and who were stilI with the university, were called after 5 years for a new title of anti-HBs and survey on anti-HBc. At that time, the subjects who had anti-HBs titles < 10 mUI/ml after having shown seroconversion with the first scheme were administered a booster dose of the vaccine (20 µ g), via 1M. After two to four weeks a new anti-HBs title was made. Of the 1,117 subjects receiving the first dose, 463 (41.5%) received the vaccine via ID and from the remaining 654 (58.5%) receiving the vaccine via 1M, 228 were further divided into a seroconversion follow-up subgroup. The prevalence ofthe Anti-HBs among the 1,117 HCW was of 10.3%. Certain professional categories or specialties have shown higher prevalences, such as laboratorians (14.4%), surgeons (16.2%) and orthopedists (16.7%). There were no anti-HBc reagent cases among anesthetists, anatomopathologists and radiologists. We have also noted higher prevalence of anti-HBc on subjects over 35 years old and on those who have been working in the health care area for more than 10 years. Among the 115 anti-HBc reagent HCW, 2 have shown to be HBsAg reagent and the remaining were positive for anti-HBs, indicating a prevalence ofHBV ofO.2%. Attendance between the first and the second doses was of 81.8% and between the second and third 79.4%. ln the intradermic group, however, absenteism was of 29.3%. VBH vaccination was safe and well tolerated, with no adverse reactions in 77.4% of the subjects. Reactions were more frequent in the ID group than in the 1M group (31.4%, 15.7%; p=0,000). ln the ID group, local reactions prevailed (hyperemia, pruritus and edema) and in the 1M group, systemic reactions (drowsiness, myalgia, nausea). There was one case of anaphylactic reaction after intradermic application of the second dose of the vaccine. After three doses of the vaccine, the seroconversion for anti-HBs was of 92.2% in the 1M group and 59.9% in the ID group. lu the 1M group, we have achieved anti-HBs < 10 mUI/ml titles in 24.5%, titles between 10 and 100 mUI/ml in 13.7% and titles above 100 mUI/ml in 61.8%. ln the ID group, these titles were found in 46.4%; 34.3% and 19.3%, respectively. Age below 30 years and being female were the factors associated with an adequate vaccinal response. Five years after the vaccinal scheme, we found titles of anti-HBs >= 10 mUI/ml in 73.7% and 43.2% ofsubjects from groups 1M and ID, respectively. After administering a booster dose in 36 subjects with anti-HBs < 10 mUI/ml, 88.9% have shown titles of anti-HBs protectors; 83.3% have shown titles above 150 mUI/m1 / Doutorado / Clinica Medica / Doutor em Clínica Médica

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2014-10-21T12:18:38Z No. of bitstreams: 3 Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 3924004 bytes, checksum: d7ba7c1ee906d0c858ef16bd5981e6f6 (MD5) Anexos - Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 6385883 bytes, checksum: fb90963b122614948d1d35bfe7554d6e (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Rejected by Luciana Ferreira (lucgeral@gmail.com), reason: Cássia, o nome do arquivo, segundo as instruções deve ser: : Dissertação - Maria Flores Silva - 2014”, “Tese - Maria Flores Silva - 2014. Você está esquecendo de colocar o traço após Tese ou Dissertação. Na citação está faltando a data final. ex. PIMENTEL, Tatiane Dias. Entre a televisão e a internet: o jornalismo e o exercício da cidadania. 2012. 150 f. Dissertação (Mestrado em Comunicação) - Universidade Federal de Goiás, Goiânia, 2012. on 2014-10-23T11:29:29Z (GMT) / Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2014-11-06T11:25:56Z No. of bitstreams: 3 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 3924004 bytes, checksum: d7ba7c1ee906d0c858ef16bd5981e6f6 (MD5) Anexos - Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 6385883 bytes, checksum: fb90963b122614948d1d35bfe7554d6e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-11-06T11:36:06Z (GMT) No. of bitstreams: 3 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 3924004 bytes, checksum: d7ba7c1ee906d0c858ef16bd5981e6f6 (MD5) Anexos - Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 6385883 bytes, checksum: fb90963b122614948d1d35bfe7554d6e (MD5) / Made available in DSpace on 2014-11-06T11:36:06Z (GMT). No. of bitstreams: 3 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 3924004 bytes, checksum: d7ba7c1ee906d0c858ef16bd5981e6f6 (MD5) Anexos - Tese Karlla Antonieta Amorim Caetano - 2014.pdf: 6385883 bytes, checksum: fb90963b122614948d1d35bfe7554d6e (MD5) Previous issue date: 2014-07-22 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Estimates indicate a total of 240 million chronic hepatitis B carriers worldwide. In Brazil, rural settlements present adverse life conditions that favor the acquisition of health problems, such as the hepatitis B virus (HBV). The objective of the present study was to investigate the epidemiology of the infections caused by the hepatitis B and D viruses in people living in settlements in the southwest region of the state of Goiás. This is an observational, analytical, cross-sectional study, with a subsequent cohort of susceptible subjects for vaccination against HBV, assessment of adherence and vaccine response. In the period from May to July of 2011, 467 subjects were selected from seven settlements in the southwest of Goiás. All subjects were interviewed and tested for the detection of the serological markers HBsAg, anti-HBc and anti-HBs, by means of the enzyme-linked immunosorbent assay (ELISA). Reactive HBsAg samples were tested for total anti-HDV and IgM, by means of the ELISA, submitted to the detection of the HBV DNA by semi-nested PCR and genotyped by sequencing. Chemiluminescent microparticle immunoassay was used for the quantitative determination of anti-HBs. The research proposal was approved by the Research Ethics Committee of the Federal University of Goiás. Of the total sample, 52.2% were men, 57.8% were married and 49% had less than five years of education. Most subjects were from cities in the midwest region (81.6%) and were older than 19 years of age (73.7%). Overall prevalence for infection by the HBV was 10.9% (51/467), being 0.8% (4/467) for HBsAg, 7.9% (37/467) for anti-HBc and anti- HBs, and 2.1% (10/467) for anti-HBc. Moreover, 19.3% of the people living in settlements presented isolated positivity for the anti-HBs marker, which indicates they were previously vaccinated against hepatitis B. The four reactive HBV DNA samples were classified as subgenotype A1 (3/4) and D3 (1/4). The male gender (adjusted OR: 2.65; p= 0.007), age (adjusted OR: 1.07; p= 0.000), history of transfusion (adjusted OR: 2.52; p= 0.025) and greater period of time living in the settlements (adjusted OR: 1.10; p= 0.026) were variables associated with the HBV. A total of 181 subjects susceptible to the infection started vaccine against hepatitis B, but only 106 (58.6%; 106/181) completed the vaccine scheme. Of these, it was possible to assess the vaccine response in 77 subjects, and 68.8% (53/77) presented protective titers of anti-HBs. Of those subjects living in the settlements who did not respond to the vaccine, a greater proportion was male, had more than 40 years and was smokers (p< 0.001). No subject presented positivity for the anti-HDV marker. The results of this study evidence the need for effective strategies to prevent hepatitis B in the studied settlements, emphasizing vaccine against hepatitis. In light of the low vaccine response against this infection in older subjects, more immunogenic schemes are suggested, so as to induce a protective vaccine response. / Estima-se em 240 milhões o número de portadores crônicos de hepatite B. No Brasil, as comunidades rurais de assentamentos apresentam condições de vida adversas que favorecem a aquisição de agravos, como do HBV. O objetivo do presente estudo foi investigar a epidemiologia das infecções pelos vírus das hepatites B e D em assentados do sudoeste de Goiás. Trata-se de um estudo observacional, analítico, de corte transversal, em que posteriormente foi formada uma coorte de indivíduos suscetíveis para a vacinação contra HBV, avaliação da adesão e resposta vacinal. No período de maio a julho de 2011, foram recrutados 467 indivíduos de sete assentamentos do sudoeste goiano. Todos foram entrevistados e testados para a detecção dos marcadores sorológicos HBsAg, anti- HBc e anti-HBs pelo ensaio imunoenzimático (ELISA). As amostras HBsAg reagentes foram testadas para o anti-HDV total e IgM, pelo ELISA, submetidas à detecção do HBV DNA por semi-nested PCR e genotipadas por sequenciamento. Para a determinação quantitativa do anti-HBs empregou-se o imunoensaio de micropartículas por quimioluminescência. O presente estudo foi aprovado pelo Comitê de Ética em Pesquisa da Universidade Federal de Goiás. Do total, 52,2% eram do sexo masculino, 57,8% casados e 49% possuía menos de cinco anos de estudo. A maioria era natural de municípios da Região Centro-Oeste (81,6%) e possuía mais de 19 anos de idade (73,7%). A prevalência global para a infecção pelo HBV foi de 10,9% (51/467), sendo 0,8% (4/467) para HBsAg, 7,9% (37/467) para anti-HBc e anti-HBs, e 2,1% (10/467) para anti-HBc. Também, 19,3% dos assentados apresentaram positividade isolada para o marcador anti-HBs, indicando vacinação prévia contra hepatite B. As quatro amostras HBV DNA reagentes foram classificadas como subgenótipo A1 (3/4) e D3 (1/4). Observou-se que sexo masculino (OR ajustado: 2,65; p= 0,007), idade (OR ajustado: 1,07; p= 0.000), história de transfusão (OR ajustado: 2,52; p= 0,025) e maior tempo de moradia nos assentamentos (OR ajustado: 1,10; p= 0,026) foram variáveis associadas ao HBV. Um total de 181 assentados suscetíveis à infecção iniciaram a vacinação contra hepatite B, mas apenas 106 (58,6%; 106/181) completaram o esquema vacinal. Desses, em 77 foi possível avaliar a resposta vacinal, sendo que 68,8% (53/77) apresentaram títulos protetores de anti-HBs. Verificou-se uma proporção maior de não respondedores nos assentados do sexo masculino, com mais de 40 anos e tabagistas (p< 0,001). Nenhum indivíduo apresentou positividade para o marcador anti-HDV. Os resultados evidenciam a necessidade de estratégias efetivas de prevenção da hepatite B nos assentamentos estudados, com ênfase na vacinação contra hepatite. Ainda, diante da baixa resposta vacinal contra esta infecção em indivíduos mais velhos, sugere-se esquemas mais imunogênicos, que induzam uma resposta vacinal protetora.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2016-09-05T13:10:46Z No. of bitstreams: 2 Dissertação - Paulie Marcelly Ribeiro dos Santos Carvalho - 2016.pdf: 5626599 bytes, checksum: 9af96621473e8d99237c7811c44ef4fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-05T13:19:07Z (GMT) No. of bitstreams: 2 Dissertação - Paulie Marcelly Ribeiro dos Santos Carvalho - 2016.pdf: 5626599 bytes, checksum: 9af96621473e8d99237c7811c44ef4fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-05T13:19:07Z (GMT). No. of bitstreams: 2 Dissertação - Paulie Marcelly Ribeiro dos Santos Carvalho - 2016.pdf: 5626599 bytes, checksum: 9af96621473e8d99237c7811c44ef4fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-02-18 / Infection caused by the hepatitis B virus (HBV) continues to have a major impact on global public health, even while being vaccine-preventable. People living on the streets, homeless, are at high risk for sexually transmitted infections (STIs), including hepatitis B. To investigate the epidemiology of HBV infection among homeless people in Goiania, Goias, from August 2014 to June 2015, 353 individuals served by a public shelter at the capital were interviewed using a structured questionnaire containing questions about socio-demographic characteristics, clinical and risk factors for HBV infection. Next, all were tested for serologic markers of HBV. It was observed the predominance of males (81.3%), mixed race selfdeclared (61%), single (59.8%), low income (70%) and low level of education (53,3%). The global prevalence of HBV was estimated at 21.81% (95% CI 17.82 to 26.41): two individuals were HBsAg / anti-HBc positive, 61 were anti-HBc / anti-HBs, and 16 showed reactivity to only the anti-HBc marker. Additionally, 19.55% (CI: 95%: from 15.75 to 24.00) tested positive for isolated anti-HBs, suggesting immunity to HBV. Analyzing potential risk factors to HBV showed that: age over 50 years, being gay or bisexual, and being mixed race/blackselfdeclared were independently associated of exposure to HBV. The results confirm the vulnerability of this populational subgroup and a high prevalence of exposure to HBV. Still. the low frequency of serological evidence of immunization against HBV, specially among older subjects, makes evident the need of drawing up strategies of vaccination at the support places to homeless. / A infecção causada pelo vírus da hepatite B (HBV) continua apresentando grande impacto na saúde pública mundial, mesmo sendo imunoprevenível. Populações em situação de rua (PSR) apresentam um risco elevado para infecções sexualmente transmissíveis (IST), incluindo a hepatite B. Para investigar a epidemiologia da infecção pelo HBV em indivíduos em situação de rua em Goiânia, Goiás, durante agosto de 2014 a junho de 2015, 353 indivíduos atendidos em um albergue público da capital foram entrevistados utilizando-se um roteiro estruturado, contendo questões sobre características sociodemograficas, clínicas e fatores de risco para a infecção pelo HBV. A seguir, todos foram testados para os marcadores sorológicos do HBV. Observou-se predomínio de indivíduos do sexo masculino (81,3%), de cor/raça parda autodeclarada (61%), solteiros (59,8%), de baixa renda (70%) e baixo nível de escolaridade (53,3%). Estimou-se uma prevalência global para o HBV de 21,81% (IC 95%: 17,82 - 26,41): dois indivíduos foram HBsAg/anti-HBc positivos, 61 foram anti-HBc/anti-HBs e 16 apresentaram reatividade do marcador anti-HBc isolado. Ainda, 19,55% (IC: 95%: 15,75 - 24,00) apresentaram positividade isolada para o anti-HBs, sugerindo imunidade para o HBV. A análise de potenciais fatores de risco para HBV mostrou que: idade > 50 anos, ser homossexual ou bissexual e ser de cor preta/negra autodeclarada foram independentemente associados a exposição ao HBV. Os resultados ratificam a vulnerabilidade desse subgrupo populacional e uma elevada prevalência de exposição ao HBV. Ainda, a baixa freqüência de evidência sorológica de imunização contra o HBV, especialmente nos indivíduos mais velhos, evidencia a necessidade de elaboração de estratégias de vacinação nos locais de apoio a PSR.

The immune system will maintain the integrity of the organism from harmful non-malicious recognizes and protects the body against exo- and endogenous toxic substances and together with the nervous and endocrine system are among regulatory systems of the organism. Recently the evidence has supported the emerging concept of different B cell subpopulations to play a direct or indirect role in a pathogenesis of spectrum of disorders. However, so far the knowledge has been limited mainly in the way of how the specific differentiation stages of B lymphocytes are involved in pathogenesis of diseases and how course of disease, stage, and eventually different treatment can affect B cell homeostasis. This work is focused on the study of peripheral CD27+ B lymphocytes (one of the least explored human B lymphocytes) in healthy controls and patients with various immunopathologies, in this case we present representative results for patients with inflammatory bowel disease. Using polychromatic flow cytometry we examined 31 of peripheral blood samples, including 14 controls, 7 patients with Crohn's disease (CD) and 5 with ulcerative colitis (UC). In 6 patients with CD, we were able to perform immunophenotyping also 2 hours after intravenous administration of infliximab, and in one patient 14 days after drug...

Recherche la Higgs boson produit en associe avec a Z boson et desintegre a paire de b-quarks. Cet etude est realise avec l'experience ATLAS aupres du LHC. Cet recherche contient le travail de l'identification de b-jets au detecteur d'ATLAS, l'etude de triggers de l'energie manquant et introduire un nouveau trigger qui permet de gagner plus de significance de signaux. La recherche de Higgs boson est poursuit avec l'implementation de nouveau trigger et faire d'analyse statistique pour estimer la significance gagne en comparaison avec l'analyse qui utilise triggers nominaux. / Search for the Higgs poson produced in association with a Z boson and decaying to a pair of b-quarks. This study is implemented with the ATLAS experience at LHC. This research includes the work on identification of b-jets in ATLAS detector, study of triggers of missing transverse energy and introduce a new trigger which is expected to gain more signal significance. The search for the Higgs boson is followed with the implementation of the new trigger and do the statistical analysis to estimate the significance gain in comparison with the analysis which use the nominal triggers.

M.Tech. (Homoeopathy) / According to the World Health Organisation (2008), an estimated two billion individuals globally, are infected with Hepatitis B (HBV). South Africa reported 864 notified new cases between 2001 and 2004 (Department Of Health, 2005), with an estimated 3-4 million chronic HBV infected black South Africans (Kew, 2008). Kwa-Zulu Natal and Free State were the most affected provinces; while 20-39 years was the most affected age group as of 2005 (Department Of Health, 2005). Workers in the health industry, intravenous drug users and children of women who have Hepatitis B are at the most risk for contracting this disease from blood products and body fluids (Immunization Action Coalition, 2007; Boon et al., 2006), where contraction of the disease can lead to liver cirrhosis, fibrosis and hepato-cellular carcinoma (Highleyman, 2008). The Expanded Program on Immunization (EPI) of the South African Department of Health (2009) suggests vaccination for Hepatitis B should be administered at six, ten and fourteen weeks, or a dose every month for 3 months. Adverse reactions associated with the vaccine include “Guillain-Barre Syndrome, arthritis, demyelinating nervous system disease” (Pratt, 2008) and anaphylaxis (Danis & Halm, 1997). Alternatives that may assist in avoiding such symptoms include: waiting until adolescence to vaccinate (Slonim et al., 2005); only vaccinating high risk groups (Francois et al., 2002); or researching an alternative (Romm, 2001). Homeoprophylaxis is the use of homeopathy to prevent the contraction or development of disease (Zoltan, 2000) and its successful use has been recorded in various disease types and locations. There have been very few studies to show the effect of individual homeopathic nosodes used as prophylactic treatment in their related diseases, with almost none of these utilising any means of serological testing (Bevan-Jones, 2009; Frost et al., 2003; Sheffield, 2006). The aim of this study was to determine the production of the antibody to the surface antigen of Hepatitis B (anti-Hbs) due to Hepatitis B 12cH nosode administration.   Forty-three participants ranging, in ages 18 to 65 years, who tested negative for the presence of anti-HBs, took part in this four week long, double-blind, placebo controlled study. Participants were randomly placed into either the Verum or Placebo group, each group receiving four lactose powders to be taken weekly for four weeks. The Active group received lactose powders medicated with Hepatitis B 12cH, whereas the Placebo group received lactose powders medicated with 96% alcohol. Participants underwent a repeat of the serum/plasma antibody testing at the conclusion of the study to determine if there were anti-HBs present in their blood. The results were then statistically analysed using nonparametric testing: Chi-squared independent test, Mann-Whitney test and Sign test. These showed that there was no change measurable effect on the surface antigen of hepatitis B (anti-HBs) of either the Verum (active medication) or Placebo group. Primary preventative medicine is becoming increasingly popular (Kuehlein et al., 2010). Both vaccination and homeoprophylaxis are examples of primary preventative medicine, where the aim is to prevent future disease. Vaccinations encourage the production of antibodies via the activation of T-helper cells and B-lymphocytes, thus providing a template for immunity against future infections (Miller, 2000; Janeway et al., 2001). While the mechanisms of vaccination are well understood, those of homeoprophylaxis are still being investigated. One theory is that nosodes enable the body to overcome diseases. Several studies have been conducted on the effects of nosodes (Bracho et al., Prophylactic vaccination against human papilloma virus infection and disease in women: a systemic review of randomized control trial.; Gosavi et al., 2012; Shuller, 2010) and have shown favourable results in the prevention of diseases associated with those homeopathic nosodes. However, only two studies have investigated the effects that nosodes have on the antibodies of the immune system (Hoover, 2006; Neustaedter, 2002) showing the need for further studies conducted in this area. The study showed that homeopathically prepared Hepatitis B 12cH nosode is not capable of eliciting an immune response that would result in the production of the antibody to the surface antigen of Hepatitis B, and thus not able to provide immunity against Hepatitis B.