Molecular mimicry between Zika virus and central nervous system inflammatory demyelinating disorders: the role of NS5 Zika virus epitope and PLP autoantigens / Mimetismo molecular entre o vírus Zika e os distúrbios inflamatórios desmielinizantes do sistema nervoso central: o papel do epítopo NS5 do vírus Zika e dos autoantígenos PLP

Abstract Background Evidence indicates a strong link between Zika virus (ZikV) and neurological complications. Acute myelitis, optic neuritis, polyneuropathy, and encephalomyelitis that mimic inflammatory idiopathic demyelination disorders (HDD) after ZikV infection have been reported in Brazil. Objective The present study aims to investigate the possible occurrence of molecular mimicry between ZikV antigens and Multiple Sclerosis (MS) autoantigens, the most frequent HDD of the central nervous system (CNS). Methods A retrospective cohort study with 305 patients admitted due to suspected arbovirus infection in Rio de Janeiro was performed, all subjects were submitted to neurological examination, and a biological sample was collected for serologic and molecular diagnostic. Bioinformatics tools were used to analyze the peptides shared between ZikV antigens and MS autoantigens. Results Of 305 patients, twenty-six were positive for ZikV and 4 presented IDD patterns found in MS cases. Sequence homology comparisons by bioinformatics approach between NS5 ZikV and PLP MS protein revealed a homology of 5/6 consecutive amino acids (CSSVPV/CSAVPV) with 83% identity, deducing a molecular mimicry. Analysis of the 3D structures revealed a similar conformation with alpha helix presentation. Conclusions Molecular mimicry between NS5 Zika virus antigen and PLP MS autoantigens emerge as a possible mechanism for IDD spectrum in genetically susceptible individuals.

Resumo Antecedentes Evidências indicam uma forte ligação entre o vírus Zika (ZikV) e complicações neurológicas. Mielite aguda, neurite óptica, polineuropatia e encefalomielite que mimetizam distúrbios inflamatórios de desmielinização idiopáticos (DDII) após infecção por ZikV têm sido relatadas no Brasil. Obejtivo O presente estudo tem como objetivo investigar a possível ocorrência de mimetismo molecular entre antígenos do ZikV e autoantígenos da Esclerose Múltipla (EM), a DDII mais frequente do sistema nervoso central (SNC). Métodos Foi realizado um estudo de coorte retrospectivo com 305 pacientes internados por suspeita de infecção por arbovirus no Rio de Janeiro, todos os indivíduos foram submetidos a exame neurológico e coleta de amostra biológica para diagnóstico sorológico e molecular. Ferramentas de bioinformática foram usadas para analisar os peptídeos compartilhados entre antígenos do ZikV e autoantígenos da EM. Resultados Dos 305 pacientes, vinte e seis foram positivos para ZikV e 4 apresentaram padrão IDD encontrado em casos de EM. As comparações de homologia de sequência por abordagem de bioinformática entre a proteína NS5 ZikV e PLP EM revelaram uma homologia de 5/6 aminoácidos consecutivos (CSSVPV/CSAVPV) com 83% de identidade, deduzindo um mimetismo molecular. A análise das estruturas 3D revelou uma conformação semelhante com apresentação em alfa-hélice. Conclusões O mimetismo molecular entre o antígeno NS5 do vírus Zika e o autoantígeno PLP da EM surge como um possível mecanismo para o espectro IDD em indivíduos geneticamente suscetíveis.

The incidence and geographical spread of SARS-CoV-2 in Rio de Janeiro, Brazil based on RT-PCR test results

Abstract INTRODUCTION Rio de Janeiro has hardly experienced coronavirus disease. METHODS Here, 87,442 reverse transcription polymerase chain reaction (RT-PCR) test results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were reported among Rio de Janeiro residents (March to September 2020). RESULTS Overall, RT-PCR positivity of 44.6% decreased over time towards 20%. Positivity was greater among males (OR=1.22; 95%CI:1.19-1.26); Black (OR=1.10; 95%CI:1.02-1.19), Brown (OR=1.16; 95%CI:1.10-1.22), and indigenous people (OR=2.11; 95%CI:0.88-5.03) compared to Whites and increased with age; with epidemic spread from the capital to inland regions. CONCLUSIONS SARS-CoV-2 keeps spreading in Rio de Janeiro, and reopening of activities may fuel the epidemic.

Identification and characterisation of SARS-CoV-2 and Human alphaherpesvirus 1 from a productive coinfection in a fatal COVID-19 case

BACKGROUND During routine Coronavirus disease 2019 (COVID-19) diagnosis, an unusually high viral load was detected by reverse transcription real-time polymerase chain reaction (RT-qPCR) in a nasopharyngeal swab sample collected from a patient with respiratory and neurological symptoms who rapidly succumbed to the disease. Therefore we sought to characterise the infection. OBJECTIVES We aimed to determine and characterise the etiological agent responsible for the poor outcome. METHODS Classical virological methods, such as plaque assay and plaque reduction neutralisation test combined with amplicon-based sequencing, as well as a viral metagenomic approach, were performed to characterise the etiological agents of the infection. FINDINGS Plaque assay revealed two distinct plaque phenotypes, suggesting either the presence of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains or a productive coinfection of two different species of virus. Amplicon-based sequencing did not support the presence of any SARS-CoV-2 genetic variants that would explain the high viral load and suggested the presence of a single SARS-CoV-2 strain. Nonetheless, the viral metagenomic analysis revealed that Coronaviridae and Herpesviridae were the predominant virus families within the sample. This finding was confirmed by a plaque reduction neutralisation test and PCR. MAIN CONCLUSIONS We characterised a productive coinfection of SARS-CoV-2 and Herpes simplex virus 1 (HSV-1) in a patient with severe symptoms that succumbed to the disease. Although we cannot establish the causal relationship between the coinfection and the severity of the clinical case, this work serves as a warning for future studies focused on the interplay between SARS-CoV-2 and HSV-1 coinfection and COVID-19 severity.

Application of the Respondent-Driven Sampling methodology in a biological and behavioral surveillance survey among female sex workers, Brazil, 2016 / Aplicação da metodologia Respondent-Driven Sampling em pesquisa biológica e comportamental com mulheres trabalhadoras do sexo, Brasil, 2016

ABSTRACT Introduction: Respondent-Driven Sampling (RDS) has been used in surveys with key populations at risk of HIV infection, such as female sex workers (FSW). This article describes the application of the RDS method among FSW in 12 Brazilian cities, during a survey carried out in 2016. Methodology: A biological and behavioral surveillance study carried out in 12 Brazilian cities, with a minimum sample of 350 FSW in each city. Tests were performed for HIV, syphilis, and hepatitis B and C infections. A social-behavioral questionnaire was also applied. Results: The sample was comprised of 4,328 FSW. For data analysis, the sample was weighted according to each participant's network size (due consideration to the implications of RDS complex design and to the effects of homophilia are recommended). Discussion: Although RDS methods for obtaining a statistical sample are based on strong statistical assumptions, allowing for an estimation of statistical parameters, with each new application the method has been rethought. In the analysis of whole-sample data, estimators were robust and compatible with those found in 2009. However, there were significant variations according to each city. Conclusion: The achieved sample size was of great relevance for assessing progress and identifying problems regarding the prevention and treatment of Sexually Transmitted Infections. New RDS studies with more time and operational resources should be envisaged. This could further network development.

RESUMO Introdução: O método de amostragem Respondent-Driven Sampling (RDS) tem sido utilizado em inquéritos com populações-chave sob maior risco de infecção pelo HIV, como as mulheres trabalhadoras do sexo (MTS). Este artigo tem o objetivo de descrever a implementação do RDS entre MTS em 12 cidades brasileiras em 2016. Metodologia: Trata-se de um estudo de vigilância biológica e comportamental realizado em 12 cidades brasileiras, com amostra mínima de 350 MTS em cada cidade. Foram realizados testes para infecções por HIV, sífilis, hepatites B e C, e aplicou-se questionário sociocomportamental. Resultados: Participaram 4.328 MTS. Para a análise dos dados, foi elaborada uma ponderação amostral considerando o tamanho da rede de cada participante; recomenda-se que o desenho complexo de amostragem por RDS e o efeito de homofilia sejam considerados. Discussão: Apesar de o RDS ser fundamentado em pressupostos estatísticos para obtenção de uma amostra probabilística e possibilitar estimação de parâmetros estatísticos, ele vem sendo repensado a cada nova aplicação. Na análise dos dados na totalidade da amostra, os estimadores mostraram-se robustos e coerentes aos encontrados em 2009. Entretanto, constataram-se grandes variações por cidade. Conclusão: O tamanho amostral alcançado foi de grande relevância para avaliar avanços e identificar problemas a respeito da prevenção e assistência às infecções sexualmente transmissíveis. Ressalta-se a necessidade de pensar estudos RDS com maior tempo e recursos para implementação, o que poderia permitir um melhor desenvolvimento das redes.

The influence of combat simulation exercises on indirect markers of muscle damage in soldiers of the brazilian army / A influência do exercício de simulação de combate sobre marcadores indiretos de lesão muscular em soldados do exército brasileiro

The combat simulation exercise is a military activity where combat activities characterized by intense physical activity are simulated. The aim of this study was to describe the behavior of indirect markers of muscle damage during military activities of combat simulation with the realization of strenuous physical efforts. This study was conducted with military recruits/volunteers (n=43; 19-24 age group) from the COMANDOS Army Corporal Training Course. The biomarkers were evaluated at the baseline (T0), 72 hours after the baseline (T1) and 63 hours after the end of the activity (T2). To compare the variables analyzed at different times, an analysis of variance (ANOVA) was applied to repeated measures with post hoc Tukey's test to identify the statistically significant differences. The criteria of significance adopted was the value of p<0,05. On average, the total body mass was significantly greater in T0 when compared to T1 (74,4kg x 69,8Kg; p< 0,05). In relation to the biochemical markers, on average, the serum levels were significantly higher (p<0,001) in T1 when compared to T0: CPK, LDH, CRP, Mb and AGPA. After 63 hours of rest, on average, the serum levels of the biochemical markers were significantly lower than in the moment T1: CPK, LDH, CRP, Mb and AGPA. However, after 63 hours of the end of the activity, all variables evaluated returned to baseline, with the exception of LDH, that on an average remained above the values observed in the moment T0. This study made possible to understand the behavior of indirect markers of muscle damage during the COMANDOS Army Corporal Training Course. However, others studies are necessary, regarding to Training Courses such as: paratrooper, mountain, jungle warfare and others to try establish a possible range of reference values for this markers during military activities.

O exercício de simulação de combate é uma atividade militar, onde as atividades de combate caracterizadas por intensa atividade física são simuladas. O objetivo deste estudo foi descrever o comportamento dos marcadores indiretos de lesão muscular durante as atividades militares de simulação de combate com a realização de esforços físicos extenuantes. Este estudo foi realizado com militares voluntários (n = 43; faixa etária = 19-24 anos) do Curso de Formação de Cabos do Exército. Os biomarcadores foram avaliados na linha de base (T0), 72 horas após o início das atividades (T1) e 63 horas após o final da atividade de (T2). Uma análise de variância (ANOVA) para medidas repetidas, com teste post hoc Tukey, foi aplicada para identificar as diferenças estatisticamente significativas nas variáveis analisadas em momentos diferentes. O critério de significância adotado foi o valor de p <0,05. Em média, a massa corporal total foi significativamente maior no T0 quando comparado a T1 (74,4kg x 69,8Kg; p <0,05). Em relação aos marcadores bioquímicos, em média, os níveis séricos foram significativamente maiores (p <0,001) em T1 quando comparados com T0 para: CPK, LDH, CRP, Mb e AGPA. Após 63 horas de descanso, em média, os níveis séricos dos marcadores bioquímicos (CPK, LDH, CRP, Mb e AGPA ) foram significativamente menores do que no momento T1. No entanto, após 63 horas a partir do fim da atividade, todas as variáveis avaliadas retornaram à linha de base, com a exceção do LDH, que, em média, manteve-se significativamente maior do que os valores observados no momento T0. Este estudo possibilitou compreender o comportamento dos marcadores indiretos de lesão muscular durante o Curso de Formação de Cabos COMANDOS. No entanto, outros estudos são necessários, com relação a cursos de formação, tais como: paraquedistas, montanha, guerra na selva, e outros, para tentar estabelecer uma possível faixa de valores de referência para estes marcadores durante as atividades militares.

Estimation of HIV incidence in two Brazilian municipalities, 2013 / Estimação da incidência de HIV em dois municípios brasileiros, 2013

ABSTRACT OBJECTIVE To estimate HIV incidence in two Brazilian municipalities, Recife and Curitiba, in the year of 2013. METHODS The method for estimating incidence was based on primary information, resulting from the Lag-Avidity laboratory test for detection of recent HIV infections, applied in a sample of the cases diagnosed in the two cities in 2013. For the estimation of the HIV incidence for the total population of the cities, the recent infections detected in the research were annualized and weighted by the inverse of the probability of HIV testing in 2013 among the infected and not diagnosed cases. After estimating HIV incidence for the total population, the incidence rates were estimated by sex, age group, and exposure category. RESULTS In Recife, 902 individuals aged 13 years and older were diagnosed with HIV infection. From these, 528 were included in the study, and the estimated proportion of recent infections was 13.1%. In Curitiba, 1,013 people aged 13 years and older were diagnosed, 497 participated in the study, and the proportion of recent infections was 10.5%. In Recife, the estimated incidence rate was 53.1/100,000 inhabitants of 13 years and older, while in Curitiba, it was 41.1/100,000, with male-to-female ratio of 3.5 and 2.4, respectively. We observed high rates of HIV incidence among men who have sex with men, of 1.47% in Recife and 0.92% in Curitiba. CONCLUSIONS The results obtained in the two cities showed that the group of men who have sex with men are disproportionately subject to a greater risk of new infections, and indicate that strategies to control the spread of the epidemic in this population subgroup are essential and urgent.

RESUMO OBJETIVO Estimar a incidência de HIV em dois municípios brasileiros, Recife e Curitiba, no ano de 2013. MÉTODOS O método de estimação da incidência foi baseado em informações primárias, resultantes do ensaio laboratorial Lag-Avidity para detecção de infecções recentes do HIV, aplicado em uma amostra dos casos diagnosticados nas duas cidades em 2013. Para a estimação da incidência de HIV para a população total das cidades, as infecções recentes detectadas na pesquisa foram anualizadas e ponderadas pelo inverso da probabilidade de teste de HIV no ano de 2013 entre os casos infectados e não diagnosticados. Após a estimação da incidência de HIV para a população total, foram estimadas as taxas de incidência por sexo, faixa de idade e categoria de exposição. RESULTADOS Em Recife, foram diagnosticados 902 indivíduos de 13 anos e mais com infecção de HIV. Desses, 528 foram incluídos no estudo, e a proporção estimada de infecções recentes foi de 13,1%. Em Curitiba, foram diagnosticadas 1.013 pessoas de 13 anos e mais, 497 participaram do estudo, e a proporção de infecções recentes foi de 10,5%. Em Recife, a taxa de incidência estimada foi de 53,1 por 100 mil habitantes de 13 anos e mais, enquanto em Curitiba, de 41,1 por 100 mil, com razão do sexo masculino para o feminino de 3,5 e 2,4, respectivamente. Foram evidenciadas elevadas taxas de incidência de HIV entre homens que fazem sexo com homens, de 1,47% em Recife e 0,92% em Curitiba. CONCLUSÕES Os resultados obtidos nas duas cidades mostraram que o grupo dos homens que fazem sexo com homens está desproporcionalmente sujeito ao maior risco de novas infecções, e indicam que estratégias para controle da disseminação da epidemia nesse subgrupo populacional são essenciais e urgentes.

Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20 percent. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24 percent) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

Evaluation of three methods for hemoglobin. Measurement in a blood donor setting

Context: The hemoglobin (HB) level is the most-used parameter for screening blood donors for the presence of anemia, one of the most-used methods for measuring Hb levels is based on photometric detection of cyanmetahemoglobin, as an alternative to this technology, Hemo Cue has developed a photometric method based on the determination of azide metahemoglobin. Objective: To evaluate the performance of three methods for hemoglobin (Hb) determination in a blood bank setting. Design: Prospective study utilizing samples to compare methods for Hb determination. Setting: Hemotherapy Service of the Hospital Israelita Albert Eistein, a private institution in the tertiary health care system. Samples: Seria blood samples were collected from 259 individuals during the period from March to June 1996. Main Measurements: Test performances and their comparisions were assessed by the analysis of coefficients of variation (CV), linear regression and mean differences. Results: The CV for the three methods were: Coulter 0.68 per cent, Cobas 0.82 per cent and Hemo Cue 0.69 per cent. There was no difference between the mean Hb determination for the three methods (p>0.05). The Coulter and Cobas methods showed the best agreement and the HemoCue method gave a lower Hb determination when compared to both the Coulter and Cobas methods. However, paris of methods involving the Hemo Cue seem to have narrower limits of agreement (+0.78 and + 1.02) than the Coulter and Cobas combination (+ 1.13). Conclusion: The three methods provide good agreement for hemoglobin determination.