Synergistic effect of the homologous PB1–NA gene constellation in Influenza A virus reassortants: Evaluation and characterization of reassortant influenza variant viruses cross-resistant to influenza Neuraminidase Inhibitors

This project aims to elucidate the functional impact of specific mutation induced changes in the NA and PB1 genes on the viral fitness and cross-resistance profile of A(H1N1)pdm09 viruses.info:eu-repo/semantics/publishedVersio

Stresse crónico e a imunidade à vacina contra a gripe em profissionais de saúde

A B S T R A C T - Introduction: Chronic stress can influence immune response to vaccines. Healthcare workersare exposed to stressors and biological hazards, the health effects of which may be preventedthrough vaccination.Objectives: This study aims to evaluate the association between stress in nurses and: (1)insufficient response to influenza vaccine, assessed one month after vaccination (T1); (2)the drop in haemagglutination-inhibition (HAI) antibodies (ab) six months after vaccination(T6).Methods: A nested case–control study was carried out with 136 healthy hospital nurses.Individual interviews, the General Health Questionnaire (GHQ12) and Maslach Burnout Inventory(MBI-HSS) were applied in order to determine the presence of stress, using the triangulationmethod at the beginning of the study (T0). Influenza vaccine was administered and titresof HAI above each strain composing influenza vaccine before vaccination (T0), at T1and T6were assessed.Results: There was no statistically relevant (5%) relationship between stress and the insuffi-cient immune response to the vaccine at T1. Nevertheless, there was an association betweenstress and the drop in HAI ab AH1at T6, when we assessed stress by the triangulation methodusing an interview (p = 0.006), GHQ12(p = 0.045) and combination of criteria (p = 0.001), evenafter multivariate analysis (respectively, p = 0.01, p < 0.05 and p = 0.002). The odds ratios were,respectively, 3.64, 2.73 and 5.22.Conclusions: The association we found, between chronic stress and the drop in HAI ab atT6, corroborates the hypothesis that stress can negatively influence immune response.Therefore, it seems reasonable to consider this issue when we implement vaccinationprogrammes for healthcare workers.R E S U M O - Introdução: O stresse crónico pode influenciar a resposta imunitária à vacinação. Os profissionais de saúde estão expostos a stressores de natureza profissional e ainda a agentes biológicos cujos efeitos poderão ser prevenidos pela vacinação. Objetivos: Estudar a associação entre a presença de stresse e (1) a “insuficiente” resposta imunitária à vacina contra a gripe, avaliada um mês após a vacinação (T1); (2) a redução dos títulos de anticorpos dirigidos às hemaglutininas (HAI) seis meses após a vacinação (T6). Métodos: Realizou-se um estudo caso-controlo incorporado num estudo de coortes com a participação de 136 enfermeiros hospitalares saudáveis. Realizaram-se entrevistas individuais e aplicaram-se os questionários The General Health Questionnaire (GHQ12) e Maslach Burnout Inventory (MBI-HSS) para determinação da presença de stresse crónico pelo método da triangulação, no início do estudo (T0). Foi administrada a vacina contra a gripe e determinou-se os títulos de HAI dirigidos a cada estirpe componentes da vacina contra a gripe, antes da vacinação(T0), em T1 e em T6. Resultados: Não se encontrou associação significativa (5%) entre a presença de stress e a “insuficiente” resposta à vacina contra a gripe em T1. Contudo, encontrou-se uma associação entre a presença de stress e a diminuição do título de HAI dirigidos à estirpe A(H1N1) em T6 quando se avaliou a presença de stresse pelo método da triangulação usando a entrevista (p=0,006), o GHQ12 (p=0,045) e a combinação dos três critérios (p=0,001), que se manteve após análise multivariada (respetivamente p=0,01, p<0.05 e p=0.002). Os odds ratio ajustados foram de 3,64, de 2,73 e de 5,22. Conclusões: A associação encontrada entre a presença de stresse crónico e a redução do título de HAI em T6 vem apoiar a hipótese de que o stresse poderá influenciar negativamente a manutenção dos títulos de anticorpos, mesmo em indivíduos adultos não idosos. Assim, parece razoável considerar este aspeto quando se pretende implementar programas de vacinação dirigidos a profissionais de saúde.info:eu-repo/semantics/publishedVersio

Stresse crónico e a imunidade à vacina contra a gripe em profissionais de saúde

A B S T R A C T - Introduction: Chronic stress can influence immune response to vaccines. Healthcare workersare exposed to stressors and biological hazards, the health effects of which may be preventedthrough vaccination.Objectives: This study aims to evaluate the association between stress in nurses and: (1)insufficient response to influenza vaccine, assessed one month after vaccination (T1); (2)the drop in haemagglutination-inhibition (HAI) antibodies (ab) six months after vaccination(T6).Methods: A nested case–control study was carried out with 136 healthy hospital nurses.Individual interviews, the General Health Questionnaire (GHQ12) and Maslach Burnout Inventory(MBI-HSS) were applied in order to determine the presence of stress, using the triangulationmethod at the beginning of the study (T0). Influenza vaccine was administered and titresof HAI above each strain composing influenza vaccine before vaccination (T0), at T1and T6were assessed.Results: There was no statistically relevant (5%) relationship between stress and the insuffi-cient immune response to the vaccine at T1. Nevertheless, there was an association betweenstress and the drop in HAI ab AH1at T6, when we assessed stress by the triangulation methodusing an interview (p = 0.006), GHQ12(p = 0.045) and combination of criteria (p = 0.001), evenafter multivariate analysis (respectively, p = 0.01, p < 0.05 and p = 0.002). The odds ratios were,respectively, 3.64, 2.73 and 5.22.Conclusions: The association we found, between chronic stress and the drop in HAI ab atT6, corroborates the hypothesis that stress can negatively influence immune response.Therefore, it seems reasonable to consider this issue when we implement vaccinationprogrammes for healthcare workers.R E S U M O - Introdução: O stresse crónico pode influenciar a resposta imunitária à vacinação. Os profissionais de saúde estão expostos a stressores de natureza profissional e ainda a agentes biológicos cujos efeitos poderão ser prevenidos pela vacinação. Objetivos: Estudar a associação entre a presença de stresse e (1) a “insuficiente” resposta imunitária à vacina contra a gripe, avaliada um mês após a vacinação (T1); (2) a redução dos títulos de anticorpos dirigidos às hemaglutininas (HAI) seis meses após a vacinação (T6). Métodos: Realizou-se um estudo caso-controlo incorporado num estudo de coortes com a participação de 136 enfermeiros hospitalares saudáveis. Realizaram-se entrevistas individuais e aplicaram-se os questionários The General Health Questionnaire (GHQ12) e Maslach Burnout Inventory (MBI-HSS) para determinação da presença de stresse crónico pelo método da triangulação, no início do estudo (T0). Foi administrada a vacina contra a gripe e determinou-se os títulos de HAI dirigidos a cada estirpe componentes da vacina contra a gripe, antes da vacinação(T0), em T1 e em T6. Resultados: Não se encontrou associação significativa (5%) entre a presença de stress e a “insuficiente” resposta à vacina contra a gripe em T1. Contudo, encontrou-se uma associação entre a presença de stress e a diminuição do título de HAI dirigidos à estirpe A(H1N1) em T6 quando se avaliou a presença de stresse pelo método da triangulação usando a entrevista (p=0,006), o GHQ12 (p=0,045) e a combinação dos três critérios (p=0,001), que se manteve após análise multivariada (respetivamente p=0,01, p<0.05 e p=0.002). Os odds ratio ajustados foram de 3,64, de 2,73 e de 5,22. Conclusões: A associação encontrada entre a presença de stresse crónico e a redução do título de HAI em T6 vem apoiar a hipótese de que o stresse poderá influenciar negativamente a manutenção dos títulos de anticorpos, mesmo em indivíduos adultos não idosos. Assim, parece razoável considerar este aspeto quando se pretende implementar programas de vacinação dirigidos a profissionais de saúde.info:eu-repo/semantics/publishedVersio

Influenza activity 2000/2001

As infecções por vírus influenza são uma importante causa de morbilidade em todos os grupos etários e estão associados a uma elevada mortalidade nos idosos e nos indivíduos pertencentes a grupos de risco. No presente estudo analisaram-se os dados da vigilância epidemiológica da gripe durante o Inverno de 2000/2001. Os dados clínicos, epidemiológicos e virológicos referentes aos casos de síndroma gripal foram recolhidos através do Programa Nacional de Vigilância da Gripe que se desenvolve em colaboração com a Direcção Geral da Saúde e integra a informação obtida a partir das redes Médicos-Sentinela e Serviços de Urgência. A análise dos dados recolhidos mostram que, durante a época de Inverno de 2000/2001, a actividade gripal foi baixa, sendo o período epidémico curto e de pequena intensidade e duração. As taxas de incidência do síndrome gripal subiram acima da linha de base durante três semanas e não ultrapassaram os 74 casos por 100 000 habitantes. Os vírus influenza do tipo B foram predominantes, verificando-se a presença simultânea de vírus influenza tipo AH1 e AH3. A caracterização antigénica e genética das estirpes isoladas permitiu confirmar a semelhança entre estas estirpes virais e as estirpes vacinais e detectar a extensão dos drifts antigénicos. Saliente-se que apesar da maioria das estirpes de vírus influenza B serem idênticas antigenicamente à estirpe vacinal, a caracterização genética mostrou uma evolução dirigida para a estirpe B/ Sichuan/379/99 que viria a integrar a vacina antigripal em 2001/2002. Consequentemente, observou-se a co-circulação de duas linhagens diferentes evidenciada pela análise filogenética das estirpes B isoladas no nosso país

Outbreak of acute respiratory infection among infants in Lisbon, Portugal, caused by human adenovirus serotype 3 and a new 7/3 recombinant strain

Human adenoviruses (AdVs) typically cause mild illnesses in otherwise healthy hosts. We investigated a pediatric outbreak of acute respiratory infection with fatal outcomes that occurred in Lisbon, Portugal, in 2004.Biological specimens were collected from 83 children attending two nurseries, a kinesiotherapy clinic, and the household of a nanny. Adenovirus infection was confirmed in 48 children by PCR and virus isolation. Most(96%) isolates were classified as being of subspecies B1. Phylogenetic analysis of fiber and hexon gene sequences revealed that most infants were infected with AdV serotype 3 (AdV3) strains. Infants attending one nursery harbored a new recombinant strain containing an AdV serotype 7 hexon and serotype 3 fiber (AdV7/3). Both the AdV3 and the AdV7/3 strains caused fatal infections. Two different serotype 3 strains were circulating in Lisbon in 2004, and the new AdV7/3 recombinant type originated from only one of those strains. These results demonstrate that recombination leads to the emergence of new adenovirus strains with epidemic and lethal potentialThis research was funded by DGE of the European Commission (for the research project entitled Genomic inventory, forensic markers, and assessment of potential therapeutic and vaccine targets for viruses relevant in biological crime and terrorism, grant SSPE-CT-2005-022639 RIVIGENE

Excess Mortality Associated with Influenza Epidemics in Portugal, 1980 to 2004

Influenza epidemics have a substantial impact on human health, by increasing the mortality from pneumonia and influenza, respiratory and circulatory diseases, and all causes. This paper provides estimates of excess mortality rates associated with influenza virus circulation for 7 causes of death and 8 age groups in Portugal during the period of 1980–2004.We compiled monthly mortality time series data by age for all-cause mortality, cerebrovascular diseases, ischemic heart diseases, diseases of the respiratory system, chronic respiratory diseases, pneumonia and influenza. We also used a control outcome, deaths from injuries. Age- and cause-specific baseline mortality was modelled by the ARIMA approach; excess deaths attributable to influenza were calculated by subtracting expected deaths from observed deaths during influenza epidemic periods. Influenza was associated with a seasonal average of 24.7 all-cause excess deaths per 100,000 inhabitants, approximately 90% of which were among seniors over 65 yrs. Excess mortality was 3–6 fold higher during seasons dominated by the A(H3N2) subtype than seasons dominated by A(H1N1)/B. High excess mortality impact was also seen in children under the age of four years. Seasonal excess mortality rates from all the studied causes of death were highly correlated with each other (Pearson correlation range, 0.65 to 0.95, P<0.001) and with seasonal rates of influenza-like-illness (ILI) among seniors over 65 years (Pearson correlation rho>0.64, P<0.05). By contrast, there was no correlation with excess mortality from injuries.Our excess mortality approach is specific to influenza virus activity and produces influenza-related mortality rates for Portugal that are similar to those published for other countries. Our results indicate that all-cause excess mortality is a robust indicator of influenza burden in Portugal, and could be used to monitor the impact of influenza epidemics in this country. Additional studies are warranted to confirm these findings in other settings

Enhanced In Vitro Antiviral Activity of Hydroxychloroquine Ionic Liquids against SARS-CoV-2

The authors also thank Fundação para a Ciência e Tecnologia for the projects Research 4 COVID-19 no. 582, and RECI/BBBBQB/0230/2012. The authors also acknowledge the Solchemar company. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.The development of effective antiviral drugs against SARS-CoV-2 is urgently needed and a global health priority. In light of the initial data regarding the repurposing of hydroxychloroquine (HCQ) to tackle this coronavirus, herein we present a quantitative synthesis and spectroscopic and thermal characterization of seven HCQ room temperature ionic liquids (HCQ-ILs) obtained by direct protonation of the base with two equivalents of organic sulfonic, sulfuric and carboxylic acids of different polarities. Two non-toxic and hydrophilic HCQ-ILs, in particular, [HCQH2 ][C1 SO3 ]2 and [HCQH2 ][GlcCOO]2, decreased the virus-induced cytopathic effect by two-fold in comparison with the original drug, [HCQH2 ][SO4 ]. Despite there being no significant differences in viral RNA production between the three compounds, progeny virus production was significantly affected (p < 0.05) by [HCQH2 ][GlcCOO]2 . Overall, the data suggest that the in vitro antiviral activities of the HCQ-ILs are most likely the result of specific intra-and intermolecular interactions and not so much related with their hydrophilic or lipophilic character. This work paves the way for the development of future novel ionic formulations of hydroxychloroquine with enhanced physicochemical properties.publishersversionpublishe

Outbreak of acute respiratory infection among infants in Lisbon, Portugal, caused by human adenovirus serotype 3 and a new 7/3 recombinant strain

Human adenoviruses (AdVs) typically cause mild illnesses in otherwise healthy hosts. We investigated a pediatric outbreak of acute respiratory infection with fatal outcomes that occurred in Lisbon, Portugal, in 2004. Biological specimens were collected from 83 children attending two nurseries, a kinesiotherapy clinic, and the household of a nanny. Adenovirus infection was confirmed in 48 children by PCR and virus isolation. Most (96%) isolates were classified as being of subspecies B1. Phylogenetic analysis of fiber and hexon gene sequences revealed that most infants were infected with AdV serotype 3 (AdV3) strains. Infants attending one nursery harbored a new recombinant strain containing an AdV serotype 7 hexon and serotype 3 fiber (AdV7/3). Both the AdV3 and the AdV7/3 strains caused fatal infections. Two different serotype 3 strains were circulating in Lisbon in 2004, and the new AdV7/3 recombinant type originated from only one of those strains. These results demonstrate that recombination leads to the emergence of new adenovirus strains with epidemic and lethal potential

Genetic variants of the IFITM3 gene reveal a potential modifier of influenza severity

As epidemias de gripe representam um sério problema de saúde pública com elevados custos económicos. Foi sugerido que um dos alelos do gene IFITM3 (rs12252-C) constitui um impor tante fator de risco de base populacional para as formas graves da gripe por infeção pelo vírus A(H1N1)pdm09. Analisámos variantes do IFITM3 associadas à gravidade da doença em doentes por tugueses (n=41). Foram identificados sete SNPs no amplicão de 352bp do IFITM3 em torno do rs12252. De acordo com HapMap, o SNP rs34481144 per tence ao mesmo bloco de desequilíbrio de ligação (LD) que rs12252, e está em LD com rs6421983. A associação negativa com formas sintomáticas graves em relação a ligeiras obser vada para o alelo rs34481144-A é sugestiva de um efeito protetor no modelo de hereditariedade dominante. Para além disso, o haplótipo Hap4 com rs34481144-A, e sem o rs12252-C, mostrou estar significativamente associado a gripe sintomaticamente ligeira. Por outro lado, apesar de a um nível de significância limiar, o haplótipo Hap1 com rs34481144-G, sem rs12252-C, mostrou-se associado à gripe com sintomatologia grave. Em comparação com a população por tuguesa em geral, foram obser vadas diferenças significativas nas frequências do alelo possivelmente protetor rs34481144-A no grupo com sintomatologia grave, do Hap1 deletério no grupo com sintomatologia ligeira e do Hap4 protetor no grupo com doença grave. A proporção de casos com sintomas graves que poderiam ser evitados se todos os indivíduos da população apresentassem o alelo protetor rs34481144-A foi estimada em 56% e 64%, respetivamente na população em geral e no grupo de indivíduos com doença ligeira. A implicação destas variantes nos fenótipos da doença necessita de estudos de validação, nomeadamente de natureza funcional.Influenza epidemics are a serious global public health and economic problem. The IFITM3 allele (rs12252-C) was suggested as a strong population-based genetic risk factor for severe influenza virus infection by A(H1N1)pdm09. We analyzed IFITM3 variants for association to influenza severity in Por tuguese patients (n=41). Seven SNPs, within the 352bp IFITM3 amplicon around rs12252, were identified. According to HapMap SNP rs34481144 belongs to the same linkage disequilibrium (LD) block as rs12252, and is in strong LD with rs6421983. A negative association with severe relative to mild disease was obser ved for allele rs34481144-A, indicating a protective ef fect under the dominant model. Moreover, haplotype Hap4 with rs34481144-A, not including rs12252-C, was significantly associated to mild influenza. Conversely, although with borderline significance, haplotype Hap1 with rs34481144-G, not including rs12252-C, was associated to severe disease. Moreover, in comparison to the general Por tuguese population, statistical significant dif ferences in the frequencies of the protective allele rs34481144-A in the severe disease group, the deleterious Hap1 in the mild disease group and the protective Hap4 in the severe disease group, were obser ved. The population attributable risk (PAR) for the targeted rs34481144 allele or genotype was of 55.91% and 64.44% in the general population and the mildly infected individuals, respectively. Implication of these variants in disease phenotype needs fur ther validation, namely through functional analysis.Este trabalho foi financiado pela Fundação Luso-Americana para o Desenvolvimento (LACR Award program - 2007) e pela Fundação para a Ciência e Tecnologia (FCT), Portugal, (POCTI/ESP/44826/2002), com a comparticipação dos fundos da Comunidade Europeia (FEDER).info:eu-repo/semantics/publishedVersio

How to dissect viral infections and their interplay with the host-proteome by immunoaffinity and mass spectrometry: A tutorial

The capabilities of bioanalytical mass spectrometry to (i) detect and differentiate viruses at the peptide level whilst maintaining high sample throughput and (ii) to provide diagnosis and prognosis for infected patients are presented as a tutorial in this work to aid analytical chemists and physicians to gain insights into the possibilities offered by current high-resolution mass spectrometry technology and bioinformatics. From (i) sampling to sample treatment; (ii) Matrix-Assisted Laser Desorption Ionization- to Electrospray Ionization -based mass spectrometry; and (iii) from clustering to peptide sequencing; a detailed step-by-step guide is provided and exemplified using SARS-CoV-2 Spike Y839 variant and the variant of concern SARS-CoV-2 Alpha (B.1.1.7 lineage), Influenza B, and Influenza A subtypes AH1N1pdm09 and AH3N2.Highlights: - Immunohistochemistry with magnetic core nanoparticles to isolate viruses. - The use of MALDI-MS for rapid virus detection is explained in detail; - The use of ESI-MS/MS to pinpoint host-patient crosstalk is explained in detail. - The absolute quantitative MS is explained for large-scale protein quantitation.This work received financial support from PT national funds (FCT/MCTES) through the projects UIDB/50006/2020 and UIDP/50006/2020 and from PROTEOMASS Scientific Society through the projects #PM001/2019 and #PM003/2016.info:eu-repo/semantics/publishedVersio