Efeito da tunicamicina sobre a replicação do vírus Mayaro em células de Aedes albopictus

Tunicamycin (TM), an inhibiting substance of bacteria, fungi and yeast proliferation, is an antibiotic isolated from Streptomyces lysosuperficus. This antibiotic inhibits the formation of N-acetylglucosamine, thereby it greatly alters the glycosilation of proteins. In this study, we found that the presence of TM causes a drastic inhibition of Mayaro virus replication in Aedes albopictus cells. Even at low concentration (0,05mg/ml), the treatment of the cells for 24 hours reduced in 50% the production of the viral infectious particles. At higher concentrations, the antibiotic reduced progressively the virus replication, resulting in 98,5% inhibition after a 48 hour treatment. The polyacrylamide gel electrophoresis analysis of [35S]-methionine-labelled proteins showed that TM at 0,05mg/ml, 0,1mg/ml e 0,15mg/ml concentration does not affect the synthesis of Mayaro virus polypeptides. Only at higher concentration (0,5mg/ml), the modification of the synthesis and of the electrophoretic mobility of viral proteins were noticed. These results suggest that, at low concentration, TM may affect Mayaro virus replication in Aedes albopictus cells through other mechanisms.Tunicamicina (TM) é um antibiótico isolado de Streptomyces lysosuperficus, caracterizado inicialmente como uma substância com atividade inibidora da proliferação de bactérias, fungos e leveduras. Este antibiótico possui um efeito bloqueador da formação de N -acetilglicosamina, alterando de maneira significativa o processo de glicosilação das proteínas. Neste trabalho, verificamos que, em células de Aedes albopictus, a presença de TM acarreta uma drástica inibição da replicação do vírus Mayaro. Mesmo em baixas concentrações de TM (0,05mg/ml), o tratamento das células durante 24 horas produz uma inibição de 50% na produção de partículas virais infecciosas. Em concentrações mais elevadas, este anti-biótico diminuiu progressivamente a replicação do vírus Mayaro, atingindo valores de 98,5% de inibição, após 48h de tratamento. A análise por eletroforese em gel de poliacrilamida das proteínas virais previamente marcadas com metionina-[35S] revela que a TM nas concentrações de 0,05 mg/ml, 0,1 mg/ml e 0,15 mg/ml não afeta a síntese dos polipeptídeos do vírus Mayaro. Apenas em concentrações mais elevadas (0,5mg/ml) observam-se alterações na síntese e na mobilidade eletroforética das glicoproteínas virais. Estes dados sugerem que, em baixas concentrações, a TM pode afetar a replicação do vírus Mayaro em células de Aedes albopictus através de outros mecanismos

Enterovirus detection in stool samples from Mozambican children with acute gastroenteritis

Funding Information: Diocreciano Bero, Ph.D. was supported by Brazilian National Council for Scientific and Technological Development and the Academy of Sciences for the Developing World (CNPq/TWAS, grant number 190,897/2015–5). The ViNaDia was sponsored by Gavi, the Vaccine Alliance through Centers for Disease Control and Prevention (CDC) , Atlanta and World Health Organization, Regional Office for Africa (WHO/AFRO), Deutsche Forschungsgemeinschaft (DFG, grant number JO369/5–1). Nilsa de Deus was fellowship of the European Foundation Initiative into African Research in Neglected Tropical Diseases (EFINTD, grant number 89,539). Publisher Copyright: © 2022Enteroviruses (EV) are predominantly enteric viruses, present in all parts of the world causing disease in humans with a broad spectrum of clinical presentations. The purpose of this study was to identify non-polio enteroviruses (NPEV) in stool samples collected from children with acute gastroenteritis (AGE) symptoms of unknown etiology in four provinces (Maputo, Nampula, Sofala and Zambézia) of Mozambique. From June 2014 to March 2018, 327 stool samples were collected from children hospitalized with AGE in health care units. NPEVs were detected in 52 samples (52/327; 15.9%) and were more frequent in children under 5 years of age. The age group from 12 to 23 months was the most affected and showed more severity of disease. We also identified 26 different EV-types with the following detection pattern EV-B>EV-C>EV-A. The major EV-types were EV-A119 (9/52; 17.3%) and EV-C99 (8/52; 15.4%), accounting for 32.7% of the total. In addition to EV-A119, other uncommon EV-types were also identified, such as EV-B75, EV-B97 and EV-C113. The current study shows a high heterogeneity of EV types circulating in children with AGE in Mozambique as well as the identification of rarely described enteroviruses.publishersversionpublishe

Molecular Characterization and Phylogenetic Study of Coxsackievirus A24v Causing Outbreaks of Acute Hemorrhagic Conjunctivitis (AHC) in Brazil

Coxsackievirus A24 variant (CA24v) is the most prevalent viral pathogen associated with acute hemorrhagic conjunctivitis (AHC) outbreaks. Sixteen years after its first outbreak in Brazil, this agent reemerged in 2003 in Brazil, spread to nearly all states and caused outbreaks until 2005. In 2009, a new outbreak occurred in the northeast region of the country. In this study, we performed a viral isolation in cell culture and characterized clinical samples collected from patients presenting symptoms during the outbreak of 2005 in Vitória, Espírito Santo State (ES) and the outbreak of 2009 in Recife, Pernambuco State (PE). We also performed a phylogenetic analysis of worldwide strains and all meaningful Brazilian isolates since 2003.Sterile cotton swabs were used to collect eye discharges, and all 210 clinical samples were used to inoculate cell cultures. Cytopathic effects in HEp-2 cells were seen in 58 of 180 (32%) samples from Vitória and 3 of 30 (10%) samples from Recife. Phylogenetic analysis based on a fragment of the VP1 and 3C gene revealed that the CA24v causing outbreaks in Brazil during the years 2003, 2004 and 2005 evolved from Asian isolates that had caused the South Korean outbreak of AHC during the summer of 2002. However, the 2009 outbreak of AHC in Pernambuco was originated from the reintroduction of a new CA24v strain that was circulating during 2007 in Asia, where CA24v outbreaks has been continuously reported since 1970.This study is the first phylogenetic analysis of AHC outbreaks caused by CA24v in Brazil. The results showed that Asian strains of CA24v were responsible for the outbreaks since 1987 and were independently introduced to Brazil in 2003 and 2009. Phylogenetic analysis of complete VP1 gene is a useful tool for studying the epidemiology of enteroviruses associated with outbreaks

Enterovirus B74 associated with hand, foot and mouth disease

Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, BraziMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Enterovírus. Ananindeua, PA, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, BraziEnterovirus 74 (EV-B74) has been associated with cases of acute flaccid paralysis (AFP) but it is not a commonly found enterovirus. In this work, we present the characterization of an EV-B74 detected from the serum sample of a one-year-old boy presenting with signs and symptoms clinically compatible with hand, foot and mouth disease (HFMD). This is the first report of EV-B74 in Brazil

Prostaglandina A1 inibe a replicação do vírus Sindbis em células de rim de macaco e em células de mosquito

The present study reports the effect of prostaglandin A1 (PGA1) on the replication of Sindbis virus in monkey kidney and mosquito cells. In PGA1 treated cells we observed a severe reduction of virus yield. In both cells lines the highest nontoxic concentration of PGA1 (10 ìg/mL) decreased virus replication, dose dependently, by more than 90%. SDS-PAGE analysis of [35S] methionine labeled proteins showed that viral proteins (E1/E2 and C) were normally synthesized in PGA1 treated Vero cells, and induction of stress proteins (HSP70 and HSP90 ) was detected in uninfected and infected cells. In Vero cells the inhibition of virus replication was accompanied by a decrease in [3H] glucosamine incorporation into the virus glycoproteins.Neste estudo nós avaliamos o efeito da prostaglandina A1 (PGA1) na replicação do vírus Síndbis em células de macaco e em células de mosquito. Nas células tratadas com PGA1 nós observamos uma redução significativa na produção de partículas virais infecciosas. Em ambas as linhagens celulares tratadas com concentrações não-tóxicas de PGA1 (1-10 µg/ml) foi observado uma redução da replicação viral, de forma dose dependente, chegando a 90% na maior dose utilizada (10 µg/ml). A análise das proteínas virais e celulares marcadas com [35S]-metionina em SDS-PAGE mostrou que as proteínas virais (E1, E2 e C) foram normalmente sintetizas em células Vero tratadas com PGA1. Além disso, observamos também a indução das proteínas de estresse (HSP70 e HSP90) nas células Vero infectadas ou não infectadas. Em células Vero foi observado uma diminuição na incorporação de [3H] glicosamina nas glicoproteínas virais

Enteroviruses associated with hand, foot, and mouth disease in Brazil

Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Enterovírus. Rio de Janeiro, RJ, Brazil

Natural circulation of human enterovirus in Maputo city, Mozambique

Submitted by sandra infurna ([email protected]) on 2016-04-24T16:41:03Z No. of bitstreams: 1 fernanda_burlandy_etal_IOC_2015.pdf: 425818 bytes, checksum: 55c629a7223867195c46cb87c6a562dd (MD5)Approved for entry into archive by sandra infurna ([email protected]) on 2016-04-24T16:53:33Z (GMT) No. of bitstreams: 1 fernanda_burlandy_etal_IOC_2015.pdf: 425818 bytes, checksum: 55c629a7223867195c46cb87c6a562dd (MD5)Made available in DSpace on 2016-04-24T16:53:33Z (GMT). No. of bitstreams: 1 fernanda_burlandy_etal_IOC_2015.pdf: 425818 bytes, checksum: 55c629a7223867195c46cb87c6a562dd (MD5) Previous issue date: 2015Ministério da Saúde. Instituto Nacional de Saúde de Moçambique. Maputo, Moçambique.Ministério da Saúde. Instituto Nacional de Saúde de Moçambique. Maputo, Moçambique.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Entovírus. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Entovírus. Rio de Janeiro, RJ, Brasil.Ministério da Saúde. Instituto Nacional de Saúde de Moçambique. Maputo, Moçambique.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Entovírus. Rio de Janeiro, RJ, Brasil.The human enteroviruses (HEV) are responsible for a large diversity of infections affecting humans. Most infections are asymptomatic, but these viruses can cause a wide spectrum of diseases, including severe cases involving the central nervous system. The aim of this study was to isolate and identify human enteroviruses in natural circulation in children less than 15 years of age admitted at the Mavalane General Hospital in Maputo City, Mozambique. In this study, 178 stool samples were processed, obtained during November 2011 to February 2012. Samples were inoculated onto cell culture and the isolates were identified as enterovirus by conventional RT-PCR in the 5’ non-coding region followed by partial VP1 sequence. Twenty-six (26) out of the 45 cell-culture positive samples were constituted by Enterovirus (14.6% of the total 178 samples). EV-29 was the serotype most prevalent. The results show the importance of maintaining the cell line Hep2C in the diagnosis and Enterovirus circulating in the Maputo city, Mozambique

Analysis of Coxsackievirus B5 Infections in the Central Nervous System in Brazil: Insights into Molecular Epidemiology and Genetic Diversity

Coxsackievirus B5 (CVB5) is one of the most prevalent enteroviruses types in humans and causes annual epidemics worldwide. In the present study, we explored viral genetic diversity, molecular and epidemiological aspects of CVB5 obtained from cerebrospinal fluid and stool samples of patients with aseptic meningitis or acute flaccid paralysis, information that is still scarce in Brazil. From 2005 to 2018, 57 isolates of CVB5 were identified in the scope of the Brazilian Poliomyelitis Surveillance Program. Phylogenetic analyses of VP1 sequences revealed the circulation of two CVB5 genogroups, with genogroup B circulating until 2017, further replaced by genogroup A. Network analysis based on deduced amino acid sequences showed important substitutions in residues known to play critical roles in viral host tropism, cell entry, and viral antigenicity. Amino acid substitutions were investigated by the Protein Variation Effect Analyzer (PROVEAN) tool, which revealed two deleterious substitutions: T130N and T130A. To the best of our knowledge, this is the first report to use in silico approaches to determine the putative impact of amino acid substitutions on the CVB5 capsid structure. This work provides valuable information on CVB5 diversity associated with central nervous system (CNS) infections, highlighting the importance of evaluating the biological impact of certain amino acids substitutions associated with epidemiological and structural analyses