Background: Chronic HBV within the Canadian Arctic is considered endemic (>2% prevalence). To control endemic rates in Nunavut, a vaccination program was initiated approximately 20 years ago, targeted at newborns and grade school students, as an interim catch-up program, such that all individuals born after 1980 are potentially vaccinated. This study investigates the efficacy of these programs and is the first seroepidemiological survey to determine HBV prevalence in Nunavut in the post-vaccination era. Methods: Anonymized serum specimens scheduled for destruction following routine medical testing were collected from individuals granting consent. Specimens were tested for antibodies to HBV (anti-HBs, anti-HBc) and hepatitis C virus. Anti-HBc positive samples were further tested for surface antigen (HBsAg) positivity, and HBV DNA was extracted from HBsAg positive samples in order to perform molecular characterization. Results: 4802 specimens were collected according to the age distribution of Nunavut, with vaccine age cohort specimens comprising just over half of all collected specimens. Overall anti-HCV+ was 0.55%, with all positivity observed among those aged 24 to 69 years old. Total anti-HBc+ prevalence was 9.40%; however, a 10-fold decrease in the rate of HBV exposure was noted among those born after 1980 compared to those born before (1.89% vs 20.1%, p<0.001). HBsAg positivity was primarily documented in individuals born before 1980 (2.55%), although cases still occurred among the vaccine age cohort (0.21%). HBV subgenotype B5 (HBV/B5), known to be unique among Inuit and Alaska Native people, was the most prevalent genotype observed (82%). Vaccine-based antibody as the sole serological marker was evident in the vaccine age cohort, although the rate of decay with increasing age was much greater than anticipated. Conclusion: Nearly two decades after the advent of HBV vaccination in Nunavut, HBV prevalence has decreased to 1.17%, indicating a non-endemic or low risk prevalence. However, the persistence of infection and a lower than expected prevalence of vaccine-based immunity in the vaccine age cohort will require further investigation to understand the causes and consequences.

Made available in DSpace on 2014-06-12T15:51:50Z (GMT). No. of bitstreams: 2 arquivo2912_1.pdf: 1186353 bytes, checksum: 92f2c91c8d664cec12729055eb4aaf14 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Fatores genéticos, imunológicos e ambientais estão envolvidos na patogênese de diversas doenças. Dentre essas influências, o sistema de histocompatibilidade (HLA) se destaca pelo seu polimorfismo e capacidade de conferir susceptibilidade ou resistência a vários distúrbios imunomediados. De acordo com o grupo étnico-racial estudado, observam-se variações no perfil HLA relacionadas a idade de surgimento, o curso clínico e resposta ao tratamento de algumas doenças (2). Os genes do HLA são os mais polimórficos dos mamíferos (2,6). É possível que o polimorfismo exista para garantir a sobrevivência das espécies (5). A herança de um haplótipo materno e paterno em co-dominância faz com que cada pessoa possa expressar em suas células diferentes moléculas do HLA, o que lhe confere a vantagem de apresentar mais peptídeos antigênicos às células T, possibilitando assim uma maior cobertura da resposta imune (7). Quanto maior o polimorfismo do HLA numa população, maior será a probabilidade de um indivíduo herdar haplótipos diferentes e, portanto, ser heterozigoto para todos os loci (7). Para a compreensão dos mecanismos de associação das moléculas de histocompatibilidade com as doenças é imprescindível conhecer a nomenclatura dos diversos componentes do MHC, os métodos pelos quais os antígenos e os alelos de histocompatibilidade são identificados, e ainda a função das moléculas de HLA no sistema imune. Assim, torna-se mais fácil o entendimento dos mecanismos pelos quais as moléculas de histocompatibilidade participam da patogenia das doenças, conferindo susceptibilidade ou proteção contra o seu desenvolvimento (2). Os produtos dos genes HLA-DRB1*, DQB1* e DPB1* são as moléculas clássicas de histocompatibilidade de classe II, que estão envolvidas na rejeição contra enxertos e na apresentação de peptídeos aos receptores dos linfócitos T (8).legais foram pesquisadas as páginas de instituições nacionais e internacionais na Internet, perfazendo um total de 52 referências. O segundo artigo, original, configura-se em estudo sobre a necessidade de entender melhor a resposta imune do organismo frente a infecção pelo HBV em amostras de doadores de sangue da Fundação HEMOPE, realizado no período de dezembro de 2008 a dezembro de 2009, com desenho de estudo do tipo caso controle. Tem como objetivo principal descrever e comparar a frequência genotípica para os alelos HLA de classe II presentes nos lócus DRB1* em doadores de sangue com a infecção pelo HBV e imunes contra esta infecção no Estado de Pernambuco

Made available in DSpace on 2014-06-12T18:28:11Z (GMT). No. of bitstreams: 2 arquivo1040_1.pdf: 1325892 bytes, checksum: f61dd605358bbaccfa20b61c802941a3 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Testes sorológicos para detecção do HBsAg e anti-HBc são realizados em doadores de sangue para evitar a transmissão da hepatite B. No entanto, a maior causa de rejeição nos hemocentros é a detecção do anti-HBc, que provoca ausência de doadores e descarte de bolsas de sangue, o que aumenta os custos nos bancos de sangue. Mas, pesquisas mostram que doadores anti-HBc positivo com ou sem o anti-HBs, podem continuar replicando o vírus e transmitir a infecção aos receptores dos hemocomponentes. Outros estudos mostram que doadores HBsAg/anti-HBc positivos na fase crônica podem conter quantidades de HBV-DNA muito baixas e não ser detectado por testes moleculares. Ainda existe um risco de transmissão que ocorre se o doador estiver na fase inicial da infecção, antes da detecção do HBsAg e que pode ser evitado pesquisando-se o HBV-DNA. Testes para detecção de ácidos nucleicos (NAT) do HIV, HCV e HBV em doadores de sangue já ocorre em alguns países, e no Brasil, está em fase de implantação para o HIV e HCV. Os objetivos deste estudo foram: pesquisar o HBV-DNA em doadores de sangue anti-HBc positivo da Fundação HEMOPE com e sem o anti-HBs, determinar a carga viral nos doadores HBsAg/anti-HBc positivo e estimar a frequência de positividade do HBV-DNA nos diferentes perfis sorológicos no período de junho de 2009 a fevereiro de 2010. Trezentos e vinte doadores anti-HBc/anti-HBs positivos, 39 HBsAg/anti-HBc positivos e 41 anti-HBc positivo isolado participaram do estudo. A pesquisa do HBV-DNA foi realizada pelo teste qualitativo de nested-PCR e a quantificação pelo teste AMPLICOR HBV MONITOR (ROCHE). Os resultados mostraram que o HBV-DNA não foi detectado em nenhum doador anti-HBc/anti-HBs positivos e anti-HBc positivo isolado, sendo quantificado em 64% dos HBsAg/anti-HBc positivos. Esses resultados sugerem que com a utilização de uma técnica mais sensível, aumentaria a possibilidade de se detectar o HBV-DNA em um número maior de doadores. Além disso, a substituição do teste do HBsAg pela detecção do HBV-DNA, requer cautela e novos estudos. No entanto, o HBV-DNA é um marcador útil para detectar a infecção na fase de janela imunológica

Indiana University-Purdue University Indianapolis (IUPUI) / Despite the lifecycle of the hepatitis B virus (HBV) being extensively investigated and described, there remains a significant gap in our knowledge of arguably one of the most crucial steps in the HBV lifecycle, the formation and maintenance of a covalently closed circular DNA (cccDNA) reservoir. Advancements in our understanding of host factors and pathways involved in cccDNA formation have been made through hypothesis driven studies and shRNA/siRNA screenings. We sought to create a targeted-unbiased assay to directly observe host factor-rcDNA interactions. This was achieved through an rcDNA Co-Immunoprecipitation paired Mass Spectrometry (rcDNA-CoIP/MS) assay. We created a DNA oligo complimentary to the open portion of the HBV rcDNA, labeled with biotin, to facilitate easy precipitation of nuclear rcDNA and complexed proteins. Proteins precipitated were analyzed through liquid chromatography paired mass spectrometry (LC/MS). Along with previously reported host factors, several factors of DNA damage repair pathways/complexes were also identified. A component of the UV-DDB complex, DDB1, surfaced as a hit. UV-DDB/rcDNA binding was confirmed through ChIP-qPCR. DDB2, the DNA damage binding component of the UV-DDB complex was knocked out in HepG2-NTCP and HepAD38 cells. This resulted in a significant decrease in the formation of cccDNA in DDB2 knockout cell lines following infection or induction. The subsequent reduction of downstream indicators of cccDNA formation such as viral RNA and proteins, HBcAg and HBeAg, showed a consistent decrease with cccDNA levels. Ectopic expression of DDB2 in the knockout cell lines rescued HBV phenotypes of cccDNA levels and its downstream indicators. Inactive mutant DDB2 plasmids were also transfected into the DDB2 K/O cell lines and failed to rescue cccDNA indicators. We therefore showed through a novel assay that we can discover novel viral rcDNA-host interactions, such as the UV-DDB complex recruiting DNA repair pathways to “repair” rcDNA to cccDNA.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19593号 / 医博第4100号 / 新制||医||1014(附属図書館) / 32629 / 京都大学大学院医学研究科医学専攻 / (主査)教授 松岡 雅雄, 教授 朝長 啓造, 教授 西渕 光昭 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

The Northern Rocky Mountains, home to the highest concentration of glaciers in the American West, are undergoing increased rates of climate warming, resulting in previously unseen ecological and hydrological outcomes. Globally, many glacier basins have experienced glacial recession to the threshold point of surpassing peak basin runoff, resulting in substantial decreases in local hydrological yield. Such findings call for models that do not alone examine glacial runoff but a complete examination of changes in the water budget. Alpine catchments are increasingly vulnerable to evapotranspirative losses due to climatic warming, and the rates of vegetation succession are often unable to keep up with the rate of warming. Basin scale analyses of glacial recession on streamflow are then confounded by ecohydrologic dynamics created by primary succession and the associated increase in evapotranspiration. In this study, I present a conceptual framework for modelling basin runoff in landscapes responding to paraglacial adjustment. The study goal was achieved by calibrating and running the Hydrologiska Byråns Vattenbalansavdelning (HBV) model in Swiftcurrent basin and investigating change across the basin water balance through baseflow analysis. The research findings indicate catchment scale changes in the timing and magnitude of the flow regime in the deglaciating Swiftcurrent basin, by employing HBV and empirical baseflow analysis. While most components of the water balance appear consistent across the study period, late summer baseflow values suggest the basin hydrology is undergoing changes, possibly a result of melt occurring earlier in the season. Ultimately, I advocate for an adaptable and accessible approach to understanding paraglacial basins by constructing an estimation of basin-scale water budgets. / Master of Science / Large scale trends in climate change are impacting a variety of ecosystems, especially alpine environments. Glacial recession has been well documented and studied in mountain chains across the globe, including the Rocky Mountains. Recession of these massive bodies of ice, which can be viewed as reservoirs of water in droughts or low flow months, has severe implications for society, the economy, and sensitive mountain environments. Furthermore, the new terrain exposed from beneath the melting glacier is dynamic and will undergo many adjustments geomorphically, in soil development, and ecologically as plants move up the glacier foreland. Ecological systems experiencing warming, deglaciation, and vegetation succession are not well understood and are complex environments due to the multiple inputs, interactions, and feedbacks. As such, this research examines how hydrologic conditions across a forty year period are changing in response to the complex feedbacks between glaciers, newly exposed terrain, and associated runoff. Through modeling and analysis, this study offers a method for understanding the water balance of Swiftcurrent basin in Glacier National Park, which can be used in other catchments experiencing similar changes.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2014-10-29T13:42:36Z No. of bitstreams: 1 Maria Isabel Magalhães Santos Perfil das....pdf: 1133942 bytes, checksum: bcc264f7cc24ec022846c05825fd31cf (MD5) / Made available in DSpace on 2014-10-29T13:42:36Z (GMT). No. of bitstreams: 1 Maria Isabel Magalhães Santos Perfil das....pdf: 1133942 bytes, checksum: bcc264f7cc24ec022846c05825fd31cf (MD5) Previous issue date: 2014 / Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Introdução: A doença causada pelo vírus da hepatite B (HBV) é um problema de saúde pública mundial. No Brasil, o sistema único de saúde (SUS) tem disponibilizado drogas antivirais para o tratamento de hepatite B crônica há mais de 10 anos, mas um sistema para o monitoramento e avaliação de resistência a estas drogas ainda não está disponível. Objetivo: Este estudo teve como objetivo determinar o perfil de mutações do HBV associadas com a resistência aos análogos de nucleos(t)ídeos entre 81 pacientes com infecção crônica pelo HBV: virgens de tratamento para hepatite B e tratados com diferentes análogos de nucleosídeos e nucleotídeos, no Hospital Professor Edgar Santos (HUPES-UFBA)- Salvador-BA. Metodologia: O HBV-DNA foi isolado de amostras de soro, amplificado por nested-PCR, utilizando-se primers deduzidos da região flaqueadora da domínio rt do gene P e sequenciados (ABI Prism 3730, Applied Biosystems, EUA). Duas a seis sequências de cada isolado foram alinhados e os sítios conflitantes foram resolvidos usando o software CLC Main Workbench v. 5.0 por inspeção visual dos eletroferogramas. As sequências consenso tinham um tamanho de 1032 pb (compreendendo os aminoácido 1-344 da rt). Estas sequências foram submetidas ao banco de dados HBVrt DB (Stanford University, EUA) para a análise de cada mutação de acordo com o genótipo e tratamento. Resultado: O genótipo A1 foi o mais prevalente (85,2%) seguido pelo genótipo A2 (4,9%) F (6,2%) e C1, D2 e D4 (1,2% cada). Seis pacientes (7 %) apresentaram mutações de resistência para LAM, ETV, TDF: dois com o padrão L180M + M204V e quatro com padrões diversos (L80I + L180M + M204I ;L80V + L180M + M204V; M204I; A194T). Todas estas mutações foram associadas ao genótipo A (quatro A1 e dois A2). Além disso, foi encontrado um paciente com HBV genótipo C típico do leste da Ásia. Destes pacientes, dois foram virgens de tratamento e quatro tinham histórico de tratamento para HIV ou HBV. Foram detectadas quatro mutações no gene S (três casos com a mutação sI195M e um a mutação sW196L) associadas às mutações do domínio rt do gene P, correspondendo à uma taxa de 6% de mutações de escape vacinal. A prevalência das mutações de resistência às drogas antivirais variou de acordo com a duração do tratamento e com o nível da barreira genética da droga utilizada. Neste estudo, foi encontrada uma forte associação entre a ocorrência de mutações de resistência do HBV e positividade para o AgHBe, co-infecção com o HIV e histórico de tratamento para HBV e/ou HIV. Conclusão: Antes da terapia ser iniciada é extremamente importante o monitoramento da carga viral e a identificação destas mutações para suportar decisões clinicas sobre o manejo dos pacientes e prevenir a emergência de vírus multi- resistentes. / Introduction: Hepatitis B virus (HBV) infection is a public health issue. The Brazilian public health system (SUS) has provided antiviral drugs for chronic hepatitis B treatment for over 10 years, but a system for monitoring for drug-related resistance mutations is not available. Objective: This study aims to determine the presence of HBV mutations associated with resistance to nucleos(t)ide analogs among 81 patients with chronic HBV infection-naïve and treated from University Hospital Professor Edgard Santos, Salvador-BA (HUPES-UFBA). Methods: Briefly, HBV-DNA was PCR amplified with primers deduced from the flanking of the rt domain at the HBV P gene and sequenced using ABI Prism 3730 (Applied Biosystems, USA). From two to six forward and reverse sequences of each isolate were assembled and conflicting sites were resolved using software CLC Main Workbench v. 5.0 by visual inspection of the electropherograms. Consensus sequence extended 1032 bp and encompassed the entire rt domain (from amino acid 1 to 344). Those sequences were submitted to the HBV drug resistance database (HBVrt DB, Stanford University, USA) to retrieve each mutation according to genotype and treatment. Results: HBV genotype A1 (85.2%) was the most prevalent followed by genotype A2 (4.9%), F (6.2%), and C1, D2 and D4 (1.2% each). Six patients (7%) exhibited resistance mutations to LAM, ETV and TDF: two with patterns L180M + M204V and four with other different patterns: L80I + L180M + M204I; L80V + L180M + M204V; M204I; A194T. All of these mutations were present in patients with genotype A (four A1 and two A2). Furthermore, this study found one patient with genotype C, common in East Asian. Of these patients, two were naïve and four had a history of treatment for HIV or HBV. In addition, four mutations in gene S (sI195M three cases with the mutation and one with the mutations W196L) associated with mutations in the rt domain of the P gene were detected, corresponding to a rate of 6% of vaccine escape mutations. Prevalence of drug-related resistance mutations varied according to treatment duration and the level of genetic barrier for the drugs used. Conclusion: In this study a strong association was found between the occurrence of HBV resistance mutations and HBeAg positivity, co-infection with HIV and a history of treatment for HBV and / or HIV. Once the drug therapy is initiated it is extremely important to monitor viral load and identify those mutations in order to support clinical decisions about patient management and also to prevent the emergence of multidrug-resistant viruses.

The Geological Survey of Sweden (SGU) has initiated a project to calibrate models to simulategroundwater levels in monitoring wells of their Groundwater Network, based on a commission fromthe Swedish Government after experiencing historically low groundwater levels and shortage in 2016and 2017. A version of the HBV model with 4 parameters, focusing on calculating groundwaterrecharge and levels, was manually calibrated to 119 groundwater stations in 2017 and the modelresults were classified according to a ‘good’, ‘poor’ or ‘bad’ visual fit to observations. In this thesis,the process-based model CoupModel, which allows the user to freely setup a model structure, wasused to simulate groundwater levels. The objectives of this thesis were to evaluate the usability of theCoupModel in groundwater level simulations and forecasting, and compare the results to previoussimulations using the HBV model. 22 groundwater stations of fast and slow responding aquifers, distributed all over Sweden, wereused to simulate groundwater levels with the CoupModel. A model structure with 11 parameters tocalibrate was constructed to represent all groundwater stations. A split-sample test was performed withcalibration of 10,000 Monte Carlo simulations and validation of the 10 simulations with the highestNash-Sutcliffe efficiency (NSE). The NSE performance was highest, and consistent through calibration and validation, for fastresponding aquifers using the CoupModel, whereas the performance of slow responding aquifers waslower. Residual analysis showed periodicity with under- and overestimations for low and highgroundwater levels, respectively, indicating that the model structure is not sufficient in representing allgroundwater stations. No relationship existed between CoupModel performance and HBV calibrationperformance, topographic position, aquifer type, location or distance to climate station. The HBVperformance was lower than for the CoupModel, with residuals of larger spread and periodicity. The CoupModel can be used for simulation and forecasting of groundwater levels, but a newmodel structure or individual structures for all groundwater stations must be constructed. A sensitivityanalysis of the parameters in the model structure must be performed to study the systematic under- andoverestimations. / Grundvatten har en stor betydelse för att upprätthålla ekosystem och försörja människor meddricksvatten, där grundvattentillgång och nivåer beror av bland annat nederbörd, temperatur,snösmältning, växtupptag och antropogen påverkan på jord och mark. Förändringar i temperatur- ochnederbördsmönster på grund av klimatförändringar och en större vattenförbrukning påverkargrundvattennivåernas variationer inom och mellan år. Den här studien syftar till att undersöka hur ochi vilken uträckning den konceptuella modellen CoupModel kan användas för simulering ochprognostisering av grundvattennivåer, samt hur den står sig i relation till en annan, tidigare studeradmodell (HBV-modellen). Studiens relevans uppdagades hos Sveriges geologiska undersökning (SGU)i samband med historiskt låga grundvattennivåer under 2016-2017, genom initiering av ett projekt medmålet att kalibrera grundvattenmodeller till mätstationer i SGUs grundvattennät. Appliceringen avmodeller har stor samhällsnytta då förebyggande av och åtgärder mot låga grundvattennivåer kanplaneras och vidtas utifrån väderprognoser och klimatscenarier. En modellstruktur skapades i CoupModel och användes för att simulera grundvattennivåer i 22grundvattenstationer av olika karaktär och modellen kalibrerades och validerades mot observationer avgrundvattennivå. Resultatet av studien visade att CoupModel kan användas som verktyg försimulering och prognostisering av grundvattennivåer, men att modellstrukturen som användes behöverutvecklas. Systematisk över- och underestimering av observerade nivåer förkommer hos allasimuleringar och ingen relation kunde ses mellan modellens prestation och plats eller typ avgrundvattenstation. CoupModel presterade i de flesta fall bättre än HBV-modellen, men kräversamtidigt mer information om en grundvattenstations jordprofil och fler parametrar att kalibrera.

Dans le cas d'une infection chronique par le virus de l'hépatite B (HBV), les traitements actuels (IFN et analogues de nucléos(t)ides) ne permettent pas d'éradiquer l'infection du fait de la persistance de l'ADNccc et des phénomènes de résistance observés chez les patients. S'agissant des traitements par l'IFN, 70 % des patients porteurs chroniques sont non-répondeurs. En effet, le virus HBV aurait développé des stratégies immunosuppressives pour établir une infection persistante. La compréhension des mécanismes impliqués dans cette viro-immunosuppression devient ainsi un enjeu majeur dans la mise en place de nouvelles stratégies antivirales. Les objectifs de ma thèse ont consisté en l'étude des relations précoces entre HBV et les hépatocytes. Nous avons pu mettre en évidence que cette absence d'activation du système immunitaire était le résultat, non pas d'une « invisibilité » du virus, mais d'une inhibition active des réponses IFN de type I/III et proinflammatoires, précocement établie par le virus HBV pour établir une infection persistante. De façon intéressante, nous avons pu démontrer que la protéine HBc était capable d'inhiber spécifiquement l'activation des voies IFN via son interaction avec les promoteurs des gènes de l'immunité innée et l'installation de marques épigénétiques (H3K9me2/3) répressives sur ces gènes par recrutement d'histone méthyl-transférases. Ces résultats prônent l'utilisation de stratégies antivirales utilisant des anticapsides dégradant et/ou prévenant la localisation nucléaire de la protéine HBc, restaurant ainsi le potentiel immunitaire des hépatocytes, pouvant dès lors être exacerbé par des agonistes de PRRs / Current treatments against Hepatitis B virus (HBV) chronic infection (IFNs and nucleos(t)ide analogues) are inefficient due to the persistence of cccDNA and emergence of viral resistance observed in infected patients. So far, up to 70 % of these patients are non responders to IFN treatments and it seems that the virus itself is able to counteract actively the host innate immune responses to establish a persistent infection. Therefore, the understanding of molecular mechanisms involved in this immunosuppression is crucial to design new immunotherapeutic strategies. In this context, the aim of my thesis was to investigate the early interactions between HBV and the hepatocyte antiviral responses. We have determined that HBV is not only a weak inducer of the host immune response, but is also able to inhibit very early and actively type I/III IFNs and proinflammatory pathways to persist in the hepatocytes. Furthermore, we have identified HBc protein as the major determinant involved specifically in the inhibition of IFN responses by counteracting host innate immune gene activations leading to repressive epigenetic marks such as H3K9/K27me3, or the recruitment of histone methyl transferase enzymes to the host IFN gene promoters. These results highlight new immunotherapeutic strategies and proposed the use of anticapsids components to degrade or block the nuclear localization of HBc proteins in order to restore a potent immune response in the hepatocytes. These anticapsid treatments may be also combined to PRRs agonists in order to improve the host antiviral state and HBV replication control

京都大学 / 0048 / 新制・課程博士 / 博士(生命科学) / 甲第20592号 / 生博第380号 / 新制||生||50(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 藤田 尚志, 教授 朝長 啓造, 教授 豊島 文子 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM