Evolutionary history and phylogeography of the hepatitis C and hepatitis B visuses in Portugal

Em 2019, a Organização Mundial de Saúde estimou 354 milhões de pessoas com hepatite B e C crónica a nível mundial. Em Portugal, o mais recente inquérito transversal a nível nacional parece revelar uma diminuição das prevalências de VHB e VHC anteriormente estimadas. A informação sobre a evolução da diversidade viral continua limitada, desconhecendo-se se a epidemiologia molecular das infeções por VHB e VHC tem vindo a ser alterada em Portugal. O objetivo deste estudo foi investigar a história evolutiva de ambos os vírus e a sua filogeografia em Portugal. Os ácidos nucleicos virais foram extraídos do plasma de doentes portugueses com VHB e VHC, os genes pol do VHB e NS5A e NS5B do VHC foram amplificados e sequenciados. A reconstrução de árvores filogenéticas de máxima verossimilhança no software IQtree v1.6.11 foi usada para genotipar as estirpes virais. A história epidémica dos subtipos do VHC em Portugal foi reconstruída através de métodos Bayesianos, conforme implementados no software BEAST v1.10.4, tal como a investigação da origem e das rotas de dispersão de ambos os vírus. As resistências aos antivirais nos dois clades do subtipo 1a do VHC foram analisadas online no Geno2Pheno [HCV] 0.92. Os nossos dados revelaram que o subgenótipo D4 do VHB foi o primeiro a ser introduzido em Portugal cerca de 1857, seguido pelo D3 e A2 algumas décadas mais tarde. Os genótipos E e A1 do VHB foram introduzidos posteriormente, quase em simultâneo. Os nossos resultados revelaram um papel muito importante de Portugal na exportação de D4 e A2 para o Brasil e Cabo Verde, respetivamente, no início do século XX. Em Portugal foram identificados subtipos distintos de VHC que entraram no país ao longo do tempo: subtipo 1b (1930-1960), subtipos 3a (1960s) e 1a (1980s), possivelmente associados a transfusões de sangue contaminado, ao início do uso de drogas intravenosas e ao seu uso generalizado, respetivamente. Os subtipos 4a e 4d, emergiram mais recentemente, possivelmente com o ressurgimento do uso de opiáceos. O subtipo 1a é claramente o mais frequente em Portugal apresentando dois clades diferentes (I e II) a circular na população, que possivelmente possuem vias de transmissão diferentes. Os primeiros países a introduzir os clades I e II em Portugal foram os Estados Unidos da América (1965) e Espanha (1955), respetivamente. Dois subclades, classificados como X e Y, foram identificados entre as estirpes do clade I. As RAS basais no gene NS5A foram encontradas principalmente nas estirpes do clade I/subclade Y, sendo a mutação mais frequente a L31M, que se revelou ausente nas estirpes do clade I/subclade X e do clade II. Este trabalho permitiu conhecer a história epidemiológica do VHB e VHC em Portugal, mostrando que Portugal teve um papel importante na dispersão global do VHB e fornecendo novos conhecimentos sobre a epidemiologia molecular, origem e dinâmica de dispersão do VHC em Portugal. Também indicou algumas estirpes virais de VHC como mais propensas a adquirir RAS e que a resistência aos antivirais deve ser investigada no contexto dos clades/subclades do subtipo 1a.In 2019, the World Health Organization estimated 354 million people with chronic hepatitis B and C worldwide. In Portugal, the most recent national cross-sectional survey indicates a decrease in previously estimated HBV and HCV prevalence. Information on the evolution of viral diversity remains limited, and it is not known whether the molecular epidemiological profile of HBV and HCV infections has been changing in Portugal. The aim of this study was to investigate the molecular epidemiology and evolutionary history of both viruses and their phylogeography in Portugal. Viral nucleic acids were extracted from the plasma of Portuguese patients infected with HBV and HCV, the pol gene of HBV and NS5A and NS5B genes of HCV were amplified and sequenced. The reconstruction of maximum likelihood phylogenetic trees in the IQtree v1.6.11 software was used for genotyping purposes. The epidemic history of HCV subtypes in Portugal was reconstructed using Bayesian methods, as implemented in the BEAST v1.10.4 software, as in the investigation of the origin and routes of spread of both viruses. Antiviral drug resistance in the two HCV subtype 1a clades were analyzed online in Geno2Pheno [HCV] 0.92. Our data indicated that the D4 subgenotype of HBV was the first to be introduced in Portugal around 1857, followed by D3 and A2 a few decades later. HBV genotypes E and A1 were introduced later, almost simultaneously. Our results revealed a very important role of Portugal in the exportation of D4 and A2 to Brazil and Cape Verde, respectively, in the beginning of the 20th century. Distinct HCV subtypes that entered Portugal over time were identified: subtype 1b (1930- 1960), subtypes 3a (1960s) and 1a (1980s), possibly associated with transfusions of contaminated blood, with the beginning of intravenous drug use and its widespread use, respectively. Subtypes 4a and 4d have emerged more recently, possibly with the resurgence of opiate use. Subtype 1a is clearly the most frequent in Portugal with two different clades (I and II) circulating in the population, which possibly have different transmission routes. The first countries to introduce clades I and II in Portugal were the United States of America Spain (1965) and Spain (1955), respectively. Two subclades, classified as X and Y, were identified among the clade I strains. The basal RAS in the NS5A gene were found mainly in the clade I/subclade Y strains, the most frequent mutation being L31M, which was absent in the strains of clade I/subclade X and clade II. This work allowed us to understand the epidemiological history of HBV and HCV in Portugal, showing that Portugal played an important role in the global spread of HBV and providing new knowledge about the molecular epidemiology, origin and dispersion dynamics of HCV in Portugal. It also highlighted that some viral strains of HCV may be more likely to acquire RAS and that antiviral resistance should be further investigated in the context of subtype 1a clades/subclades

Estresse gerado pelo trabalho em turno : estudo de caso em operadores da Pequena Central Hidrelétrica (PCH) Albano Machado - Trindade do Sul - RS

Orientador : Especialista Jonas Lopes MagalhãesMonografia (especialização) - Universidade Federal do Paraná, Setor de Tecnologia, Curso de Especialização em Engenharia de Segurança do Trabalho.Inclui referênciasResumo : No mundo globalizado, as doenças ocupacionais estão cada dia mais presentes em nosso cotidiano. A competitividade acirrada, falta de perspectivas futuras, insegurança, ansiedade e aborrecimento, induzem no trabalhador situações estressantes. É dentro desse enfoque que buscou-se mostrar os fatores de risco do estresse nesses trabalhadores. Conhecendo as causas, efeitos do estresse, suas conseqüências e tomando consciência do equilíbrio bem fundamentado, pode-se obter formas de trabalho menos estressantes e, dessa forma, reduzir a carga psicofisiológica para o trabalhador. Dando destaque para trabalhadores em Usinas Hidrelétricas, com revezamento de turnos, que tem se mostrado atuantes, o que pode causar comprometimento ao desempenho desses operadores. Este trabalho apresenta um estudo quantitativo sobre fatores que podem levar ao estresse nos operadores da Usina Hidrelétrica Albano Machado, no município de Trindade do Sul, estado do Rio Grande do Sul – RS, do Grupo Electra. Nele estão enfocados aspectos relacionados às atividades laborais dos operadores e a ocorrência do estresse. As informações foram obtidas através de roteiro de entrevista aplicada diretamente aos operadores, em seu local de trabalho. Os resultados mostram que, em sua maioria, os operadores desconhecem sobre a doença, mas de uma forma geral fornecem uma visão global dos fatores de risco relacionados ao problema estresse nos operadores da Usina albano machado. Foi recomendado que se façam modificações na parte física da usina e também que a empresa desenvolva alternativas de lazer para os funcionários e seus familiares, bem como formas de evitar a monotonia no período noturno

Antibody response against selected epitopes in the HIV-1 envelope gp41 ectodomain contributes to reduce viral burden in HIV-1 infected patients

The ectodomain of gp41 is the target of potent binding and neutralizing antibodies (NAbs) and is being explored in new strategies for antibody-based HIV vaccines. Previous studies have suggested that the W164A-3S (3S) and EC26-2A4 (EC26) peptides located in the gp41 ectodomain may be potential HIV vaccine candidates. We assessed 3S- and EC26-specific binding antibody responses and related neutralizing activity in a large panel of chronic HIV-1-infected Portuguese individuals on ART. A similar proportion of participants had antibodies binding to 3S (9.6%) and EC26 (9.9%) peptides but the level of reactivity against 3S was significantly higher compared to EC26, except in the rare patients with double peptide reactivity. The higher antigenicity of 3S was unrelated with disease stage, as assessed by CD4+ T cell counts, but it was directly related with plasma viral load. Most patients that were tested (89.9%, N = 268) showed tier 1 neutralizing activity, the potency being inversely associated with plasma viral load. In the subset of patients that were tested for neutralization of tier 2 isolates, neutralization breadth was inversely correlated with plasma viral load and directly correlated with CD4+ T cell counts. These results are consistent with a role for neutralizing antibodies in controlling viral replication and preventing the decline of CD4+ T lymphocytes. Importantly, in patients with 3S-specific antibodies, neutralizing titers were inversely correlated with viral RNA levels and proviral DNA levels. Moreover, patients with 3S and/or EC26-specific antibodies showed a 1.9-fold higher tier 2 neutralization score than patients without antibodies suggesting that 3S and/or EC26-specific antibodies contribute to neutralization breadth and potency in HIV-1 infected patients. Overall, these results suggest that antibodies targeting the S3 and EC26 epitopes may contribute to reduce viral burden and provide further support for the inclusion of 3S and EC26 epitopes in HIV-1 vaccine candidates.publishersversionpublishe

Phylogeography of hepatitis B virus: the role of Portugal in the early dissemination of HBV worldwide

Copyright: © 2022 Marcelino et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.In Portugal, the genetic diversity, origin of HBV and the Portuguese role in the dissemination of HBV worldwide were never investigated. In this work, we studied the epidemic history and transmission dynamics of HBV genotypes that are endemic in Portugal. HBV pol gene was sequenced from 130 patients followed in Lisbon. HBV genotype A was the most prevalent (n = 54, 41.5%), followed by D (n = 44, 33.8%), and E (n = 32, 24.6%). Spatio-temporal evolutionary dynamics was reconstructed in BEAST using a Bayesian Markov Chain Monte Carlo method, with a GTR nucleotide substitution model, an uncorrelated lognormal relaxed molecular clock model, a Bayesian skyline plot, and a continuous diffusion model. HBV subgenotype D4 was the first to be introduced in Portugal around 1857 (HPD 95% 1699-1931) followed by D3 and A2 a few decades later. HBV genotype E and subgenotype A1 were introduced in Portugal later, almost simultaneously. Our results indicate a very important role of Portugal in the exportation of subgenotypes D4 and A2 to Brazil and Cape Verde, respectively, in the beginning of the XX century. This work clarifies the epidemiological history of HBV in Portugal and provides new insights in the early and global epidemic history of this virus.This work was performed in the context of Rute Marcelino PhD study, whose student’s fellowship (SFRH/BD/99507/2014) was supported by the Portuguese Agency for Scientific Research, Fundação para a Ciência e Tecnologia (FCT), POCH program, Portugal 2020, and European Union/Social European Fund (FSE). This work was also supported by FCT through funds of AA's projects GHTM-UID/Multi/04413/2013 and GHTM-UID/04413/2020 and also NT's projects UIDB/04138/2020 and UIDP/04138/2020.info:eu-repo/semantics/publishedVersio

Epidemic history of hepatitis C virus genotypes and subtypes in Portugal

Any successful strategy to prevent and control HCV infection requires an understanding of the epidemic behaviour among the different genotypes. Here, we performed the first characterization of the epidemic history and transmission dynamics of HCV subtypes in Portugal. Direct sequencing of NS5B was performed on 230 direct-acting antiviral drugs (DAA)-treatment naïve patients in Lisbon. Phylogenetic analysis was used for subtyping and transmission cluster identification. Bayesian methods were used to reconstruct the epidemic history of HCV subtypes. Sequences were analysed for resistance-associated substitutions (RAS). The majority of strains were HCV-GT1 (62.6%), GT3 (18.3%, all subtype 3a) and GT4 (16.1%). Among GT1, the most frequent were subtypes 1a (75.5%) and 1b (24.5%). Polyphyletic patterns were found in all but 12 lineages suggesting multiple introductions of the different subtypes in this population. Five distinct epidemics were identified. The first significant HCV epidemic in Portugal occurred between 1930s and 1960s, was caused almost exclusively by GT1b and was likely associated with blood transfusions. Rapid expansion of GT3a occurred in the 1960s and GT1a in the 1980s, associated with intravenous drug use. The most recent epidemics were caused by GT4a and GT4d and seem to be associated with the resurgence of opioid use. The C316N substitution was found in 31.4% of GT1b-patients. Close surveillance of patients bearing this mutation and undergoing dasabuvir-based regimens will be important to determine its impact on treatment outcome.publishersversionpublishe

Characterization of NS5A and NS5B Resistance-Associated Substitutions from Genotype 1 Hepatitis C Virus Infected Patients in a Portuguese Cohort

This study is focused on the prevalent NS5 coding region resistance-associated substitutions (RASs) in DAA-naive genotype (GT)1 HCV-infected patients and their potential impact on success rates. Plasma RNA from 81 GT1 HCV-infected patients was extracted prior to an in-house nested RT-PCR of the NS5 coding region, which is followed by Sanger population sequencing. NS5A RASs were present in 28.4% (23/81) of all GT1-infected patients with 9.9% (8/81) having the Y93C/H mutation. NS5B RASs showed a prevalence of 14.8% (12/81) and were only detected in GT1b. Overall 38.3% (31/81) of all GT1 HCV-infected patients presented baseline RASs. The obtained data supports the usefulness of resistance testing prior to treatment since a statistically significant association was found between treatment failure and the baseline presence of specific NS5 RASs known as Y93C/H (p = 0.04).publishersversionpublishe

PERCEPÇÃO DE IDOSOS A RESPEITO DO BEM-ESTAR, QUALIDADE DE VIDA E SUPORTE SOCIOAMBIENTAL

Este estudo transversal teve o objetivo de verificar a percepção de 20 idosos a respeito do bem-estar subjetivo,qualidade de vida e suporte socioambiental. Participaram idosos de ambos os sexos, dos serviços de saúdeofertados por uma instituição de ensino superior. Foram utilizados a Escala de Afetos Positivos e Afetos Negativos,o Perfil de Saúde de Nottingham, e o questionário de suporte socioambiental. A análise dos dados foi realizadapor meio da estatística descritiva. Identificou-se que os idosos apresentaram melhor resultado nos domínios daqualidade de vida referentes aos aspectos subjetivos, interação social (87%) e reações emocionais (83,3%). Osvalores referentes à média total (3,37) dos afetos positivos correspondem a uma percepção de bem-estar moderada, enquanto em relação aos afetos negativos, tanto dos homens quanto das mulheres encontram-se com escores menores (1,83 e 1,94, respectivamente). Referente aos aspectos sociais e ambientais, quando analisadosseparadamente, encontrou-se uma média similar (3,75 e 3,89, respectivamente), representando uma percepçãomoderada do suporte socioambiental. Pode-se concluir que os idosos apresentaram percepção de bem-estar moderada em relação aos afetos positivos, com destaque para a determinação, entusiasmo e força, além de baixoíndice para os afetos negativos, com valores maiores para irritabilidade, nervosismo e inquietação

Grupo Projeto Creche

Esta newsletter 1 apresenta o Grupo Projeto Creche (GPC), uma comunidade de aprendizagem que nasceu no ano letivo de 2008/2009 na Escola Superior de Educação e Ciências Sociais – Instituto Politécnico de Leiria (ESECS-IPL), impulsionado pela necessidade de refletir e investigar sobre o trabalho pedagógico desenvolvido em contexto de Creche, com crianças até aos três anos de idade.info:eu-repo/semantics/publishedVersio

Day Care Project Group: The child in the day care school context

Pensando na criança até aos 3 anos, esta newsletter #2 destaca a sua essência enquanto ser humano, cidadã com direitos e deveres, num contexto social específico. Para exercer a sua cidadania necessita de um contexto relacional que dê espaço às linguagens próprias desta etapa da vida.info:eu-repo/semantics/publishedVersio