RESUMO
Hepatitis C virus (HCV) infection poses a significant public health challenge and often leads to long-term health complications and even death. Parkinson's disease (PD) is a progressive neurodegenerative disorder with a proposed viral etiology. HCV infection and PD have been previously suggested to be related. This work aimed to identify potential biomarkers and pathways that may play a role in the joint development of PD and HCV infection. Using BioOptimatics-bioinformatics driven by mathematical global optimization-, 22 publicly available microarray and RNAseq datasets for both diseases were analyzed, focusing on sex-specific differences. Our results revealed that 19 genes, including MT1H, MYOM2, and RPL18, exhibited significant changes in expression in both diseases. Pathway and network analyses stratified by sex indicated that these gene expression changes were enriched in processes related to immune response regulation in females and immune cell activation in males. These findings suggest a potential link between HCV infection and PD, highlighting the importance of further investigation into the underlying mechanisms and potential therapeutic targets involved.
Assuntos
Hepatite C , Doença de Parkinson , Feminino , Humanos , Masculino , Biomarcadores , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Hepacivirus/genética , Hepatite C/complicações , Hepatite C/virologia , Doença de Parkinson/genética , Doença de Parkinson/virologia , Fatores SexuaisRESUMO
Influenza circulation was significantly affected in 2020-21 by the COVID-19 pandemic. During this time, few influenza cases were recorded. However, in the summer of 2021-22, an increase in atypical influenza cases was observed, leading to the resurgence of influenza in the southernmost state of Brazil, Rio Grande do Sul (RS). The present study aimed to identify the circulation of FLUAV, FLUBV and SARS-CoV-2 and characterize the influenza genomes in respiratory samples using high-throughput sequencing technology (HTS). Respiratory samples (n = 694) from patients in RS were selected between July 2021 and August 2022. The samples were typed using reverse transcriptase real-time PCR (RT-qPCR) and showed 32% (223/694) of the samples to be positive for SARS-CoV-2, 7% for FLUAV (H3) (49/694). FLUBV was not detected. RT-qPCR data also resulted in FLUAV and SARS-CoV-2 co-infections in 1.7% (4/223) of samples tested. Whole genome sequencing of FLUAV produced 15 complete genomes of the H3N2 subtype, phylogenetically classified in the 3C.2a1b.2a.2a.3 subclade and revealing the dominance of viruses in the southern region of Brazil. Mutation analysis identified 72 amino acid substitutions in all genes, highlighting ongoing genetic evolution with potential implications for vaccine effectiveness, viral fitness, and pathogenicity. This study underscores limitations in current surveillance systems, advocating for comprehensive data inclusion to enhance understanding of influenza epidemiology in southern Brazil. These findings contribute valuable insights to inform more effective public health responses and underscore the critical need for continuous genomic surveillance.
Assuntos
COVID-19 , Genoma Viral , Influenza Humana , Filogenia , SARS-CoV-2 , Humanos , Brasil/epidemiologia , COVID-19/epidemiologia , COVID-19/virologia , SARS-CoV-2/genética , SARS-CoV-2/classificação , SARS-CoV-2/isolamento & purificação , Influenza Humana/epidemiologia , Influenza Humana/virologia , Pessoa de Meia-Idade , Adulto , Feminino , Genoma Viral/genética , Masculino , Adulto Jovem , Idoso , Adolescente , Surtos de Doenças , Sequenciamento Completo do Genoma , Criança , Pré-Escolar , Lactente , Coinfecção/epidemiologia , Coinfecção/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Idoso de 80 Anos ou mais , GenômicaRESUMO
Dengue is an arbovirus infection whose etiologic agent is transmitted by the Aedes aegypti mosquito. Since the early 1980s, when the circulation of the dengue virus (DENV) was confirmed in Brazil, the disease has become a growing multifactorial public health problem. This article presented the main factors that have contributed to the frequent dengue epidemics in recent years, such as the behavior of the vector, climate change, and social, political, and economic aspects. The intersection between these different factors in the dynamics of the disease is highlighted, including the increase in the mosquito population due to higher temperatures and rainy periods, as well as the influence of socioeconomic conditions on the incidence of dengue. Some mosquito control strategies are also addressed, including the use of innovative technologies such as drones and the Wolbachia bacterium, as well as the hope represented by the dengue vaccine. Nevertheless, the need for integrated and effective public policies to reduce social inequalities and the impacts of climate change on the spread of dengue is emphasized.
Assuntos
Aedes , Mudança Climática , Dengue , Mosquitos Vetores , Dengue/transmissão , Dengue/epidemiologia , Dengue/prevenção & controle , Aedes/virologia , Animais , Humanos , Brasil/epidemiologia , Controle de Mosquitos/métodos , Meio Ambiente , Fatores SocioeconômicosRESUMO
The Zika virus (ZIKV) epidemic declared in Brazil between 2015 and 2016 was associated with an increased prevalence of severe congenital malformations, including microcephaly. The distribution of microcephaly cases was not uniform across the country, with a disproportionately higher incidence in the Northeast region (NE). Our previous work demonstrated that saxitoxin (STX), a toxin present in the drinking water reservoirs of the NE, exacerbated the damaging effects of ZIKV on the developing brain. We hypothesized that the impact of STX might vary among different neural cell types. While ZIKV infection caused severe damages on astrocytes and neural stem cells (NSCs), the addition of STX did not exacerbate these effects. We observed that neurons subjected to STX exposure were more prone to apoptosis and displayed higher ZIKV infection rate. These findings suggest that STX exacerbates the harmful effects of ZIKV on neurons, thereby providing a plausible explanation for the heightened severity of ZIKV-induced congenital malformations observed in Brazil's NE. This study highlights the importance of understanding the interactive effects of environmental toxins and infectious pathogens on neural development, with potential implications for public health policies.
Assuntos
Astrócitos , Células-Tronco Neurais , Neurônios , Saxitoxina , Infecção por Zika virus , Zika virus , Células-Tronco Neurais/virologia , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , Humanos , Zika virus/fisiologia , Astrócitos/virologia , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Neurônios/virologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Infecção por Zika virus/virologia , Infecção por Zika virus/patologia , Saxitoxina/toxicidade , Apoptose/efeitos dos fármacos , Microcefalia/virologia , Morte Celular/efeitos dos fármacos , Brasil , Células CultivadasRESUMO
On 2 February 2024, the Pan American Health Organization/World Health Organization issued an epidemiological alert on rising Oropouche virus (OROV) infections in South America. By 3 August 2024, this alert level had escalated from medium to high. OROV has been a public health concern in Central and South America since its emergence in Brazil in the 1960s. However, the 2024 outbreak marks a turning point, with the sustained transmission in non-endemic regions of Brazil, local transmission in Cuba, two fatalities and several cases of vertical transmission. As of the end of August 2024, 9852 OROV cases have been confirmed. The 2024 OROV outbreak underscores critical gaps in our understanding of OROV pathogenesis and highlights the urgent need for antivirals and vaccines. This review aims to provide a concise overview of OROV, a neglected orthobunyavirus.
Assuntos
Infecções por Bunyaviridae , Orthobunyavirus , Orthobunyavirus/genética , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , Infecções por Bunyaviridae/transmissão , Humanos , Animais , Surtos de Doenças , Doenças Transmissíveis Emergentes/virologia , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/transmissão , América do Sul/epidemiologiaRESUMO
OBJECTIVE: We developed an in-house bioinformatics pipeline to improve the detection of respiratory pathogens in metagenomic sequencing data. This pipeline addresses the need for short-time analysis, high accuracy, scalability, and reproducibility in a high-performance computing environment. RESULTS: We evaluated our pipeline using ninety synthetic metagenomes designed to simulate nasopharyngeal swab samples. The pipeline successfully identified 177 out of 204 respiratory pathogens present in the compositions, with an average processing time of approximately 4 min per sample (processing 1 million paired-end reads of 150 base pairs). For the estimation of all the 470 taxa included in the compositions, the pipeline demonstrated high accuracy, identifying 420 and achieving a correlation of 0.9 between their actual and predicted relative abundances. Among the identified taxa, 27 were significantly underestimated or overestimated, including only three clinically relevant pathogens. We also validated the pipeline by applying it to a clinical dataset from a study on metagenomic pathogen characterization in patients with acute respiratory infections and successfully identified all pathogens responsible for the diagnosed infections. These findings underscore the pipeline's effectiveness in pathogen detection and highlight its potential utility in respiratory pathogen surveillance.
Assuntos
Metagenômica , Infecções Respiratórias , Metagenômica/métodos , Humanos , Infecções Respiratórias/microbiologia , Infecções Respiratórias/diagnóstico , Metagenoma/genética , Biologia Computacional/métodos , Reprodutibilidade dos Testes , Nasofaringe/microbiologia , Nasofaringe/virologiaRESUMO
Cassava (Manihot esculenta Crantz) is a vital carbohydrate source for over 800 million people globally, yet its production in East Africa is severely affected by cassava brown streak disease (CBSD). Genebanks, through ex-situ conservation, play a pivotal role in preserving crop diversity, providing crucial resources for breeding resilient and disease-resistant crops. This study genotyped 234 South American cassava accessions conserved at the CIAT genebank, previously phenotyped for CBSD resistance by an independent group, to perform a genome-wide association analysis (GWAS) to identify genetic variants associated with CBSD resistance. Our GWAS identified 35 single nucleotide polymorphism (SNP) markers distributed across various chromosomes, associated with disease severity or the presence/absence of viral infection. Markers were annotated within or near genes previously identified with functions related to pathogen recognition and immune response activation. Using the SNP candidates, we screened the world's largest cassava collection for accessions with a higher frequency of favorable genotypes, proposing 35 accessions with potential resistance to CBSD. Our results provide insights into the genetics of CBSD resistance and highlight the importance of genetic resources to equip breeders with the raw materials needed to develop new crop varieties resistant to pests and diseases.
Assuntos
Resistência à Doença , Estudo de Associação Genômica Ampla , Manihot , Doenças das Plantas , Polimorfismo de Nucleotídeo Único , Manihot/genética , Manihot/virologia , Manihot/parasitologia , Resistência à Doença/genética , Doenças das Plantas/genética , Doenças das Plantas/virologia , América do Sul , Genótipo , Genoma de Planta , PotyviridaeRESUMO
Hantavirus Pulmonary Syndrome (HPS), characterized by its high fatality rate, poses a significant public health concern in Argentina due to the increasing evidence of person-to-person transmission of Andes virus. Several orthohantaviruses were described in the country, but their phylogenetic relationships were inferred from partial genomic sequences. The objectives of this work were to assess the viral diversity of the most prevalent orthohantaviruses associated with HPS cases in the Central-East (CE) region of Argentina, elucidate the geographic patterns of distribution of each variant and reconstruct comprehensive phylogenetic relationships utilizing complete genomic sequencing. To accomplish this, a detailed analysis was conducted of the geographic distribution of reported cases within the most impacted province of the region. A representative sample of cases was then selected to generate a geographic map illustrating the distribution of viral variants. Complete viral genomes were obtained from HPS cases reported in the region, including some from epidemiologically linked cases. The phylogenetic analysis based on complete genomes defined two separate clades in Argentina: Andes virus in the Southwestern region and Andes-like viruses in other parts of the country. In the CE region, Buenos Aires virus and Lechiguanas virus clearly segregate in two subclades. Complete genomes were useful to distinguish person-to-person transmission from environmental co-exposure to rodent population. This study enhances the understanding of the genetic diversity, geographical spread, and transmission dynamics of orthohantaviruses in Central Argentina and prompt to consider the inclusion of Buenos Aires virus and Lechiguanas virus in the species Orthohantavirus andesense, as named viruses.
Assuntos
Variação Genética , Genoma Viral , Orthohantavírus , Filogenia , Argentina/epidemiologia , Orthohantavírus/genética , Orthohantavírus/classificação , Humanos , Sequenciamento Completo do Genoma , Síndrome Pulmonar por Hantavirus/transmissão , Síndrome Pulmonar por Hantavirus/virologia , Síndrome Pulmonar por Hantavirus/epidemiologia , Masculino , Feminino , Adulto , Animais , Pessoa de Meia-Idade , Infecções por Hantavirus/transmissão , Infecções por Hantavirus/virologia , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/veterinária , Adulto JovemRESUMO
This systematic review aims to determine whether the presence of human papillomavirus (HPV) influences the immunohistochemical expression of programmed cell death-1 ligand (PD-L1) in oropharyngeal squamous cell carcinoma (OPSCC). PD-L1 immunohistochemical expression varies in OPSCC, and the presence of HPV is a plausible explanation for this variability. Comprehending these findings is crucial, as high PD-L1 expression in the tumor microenvironment of OPSCC can help identify patient subgroups that could be suitable for immunotherapy. Therefore, a systematic review was conducted following PRISMA guidelines (CRD42023437800). An electronic literature search was performed without time or language restrictions. The search included PubMed/MEDLINE, Embase, Scopus, Web of Science, https://clinictrials.gov, and relevant journals. A meta-analysis was performed using RStudio. Fourteen studies involving 1,629 participants were included. The sample consisted predominantly of males (81.26%) with a mean age of 58.3 years. Concerning clinical and pathological characteristics, the most frequently described anatomical location was the tonsils (68.54%), and most participants were either current or former smokers (78%) and alcohol users (79%). Advanced TNM IV was the most common stage. Regarding histopathological characteristics, HPV 16 was the only type mentioned, and half of the cases were detected through immunohistochemistry. The SP142 clone (35.7%) and the pattern of membrane immunostaining in tumor cells (71%) were the most commonly employed methods. The most prevalent findings were positive expression of PD-L1 (64.28%) and negative HPV status (57.14%). The association between PD-L1 positivity and HPV positivity (78.57%) was confirmed by meta-analysis. The conclusion was that HPV-positive status has an impact on immunohistochemical expression of PD-L1 in OPSCC.
Assuntos
Antígeno B7-H1 , Carcinoma de Células Escamosas , Imuno-Histoquímica , Neoplasias Orofaríngeas , Infecções por Papillomavirus , Humanos , Antígeno B7-H1/análise , Neoplasias Orofaríngeas/virologia , Neoplasias Orofaríngeas/patologia , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Masculino , Feminino , Carcinoma de Células Escamosas/virologia , Carcinoma de Células Escamosas/patologia , Pessoa de Meia-Idade , Biomarcadores Tumorais/análise , PapillomaviridaeRESUMO
Antibodies are an essential component of the antiviral response in many species, but to date, there is no compelling evidence that bats are capable of eliciting a robust humoral immunity, including neutralizing antibodies. Here, we report that infection of Jamaican fruit bats with the bat influenza A virus H18N11 elicits a rapid and stable humoral immune response with a strong neutralizing capacity, associated with no detectable viral shedding after repeat challenge infection. Thus, the neutralizing antibody response of bats might play an important role in the bat immunity.
Assuntos
Anticorpos Neutralizantes , Anticorpos Antivirais , Quirópteros , Infecções por Orthomyxoviridae , Quirópteros/virologia , Quirópteros/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Infecções por Orthomyxoviridae/veterinária , Anticorpos Antivirais/imunologia , Vírus da Influenza A/imunologia , Eliminação de Partículas Virais/imunologiaRESUMO
INTRODUCTION: The discovery of the presence of the SARS-CoV-2 receptor and carrier protein in the testicles, along with the mandatory preventive social isolation during 2020 and subsequent immunization, prompted us to evaluate the effect of the COVID-19 pandemic on seminal variables in males seeking consultation at the laboratory. MATERIALS AND METHODS: An analytical and observational experimental design was employed. Seminal variables from semen analyses and kinetic values were analyzed using a computer-assisted sperm analysis system in 409 semen samples collected from patients attending the laboratory between April and December 2019, and April and December 2021. COVID-positive patients were stratified based on the time elapsed since the illness into three groups: less than 3 months (acute phase), 4-12 months, and more than 1 year. RESULTS: A significant difference (p=0.013) was found in the total sperm count per ejaculate in the COVID-positive group compared to the prepandemic and COVID-negative groups (Median (Q1-Q3): 67.49 (26.42-139.44) vs. 102.48 (43.86-197.05) and 96.72 (38.22-189.27)). When stratifying the COVID-positive group based on the time since the illness, the significant decrease (p=0.038) occurred during the acute phase, with recovery over time to values similar to the prepandemic and COVID-negative groups. Social isolation and vaccination did not have significant effects on seminal variables. DISCUSSION: The study revealed reversible changes in testicular function reflected by a decrease in sperm count in the total ejaculate of patients who had experienced COVID-19. These changes appear to be related to fever and inflammation rather than the virus infection itself.
Introducción: El hallazgo en testículo del receptor y la proteína transportadora del virus SARS-CoV-2, el aislamiento social preventivo obligatorio durante 2020 y la inmunización, nos condujeron a evaluar el efecto de la pandemia COVID-19 sobre las variables seminales de varones que consultaron al laboratorio. Materiales y métodos: Diseño experimental analítico, observacional y retrospectivo. Se analizaron las variables seminales del espermograma y los valores cinéticos mediante un sistema computarizado, en 409 muestras de semen de pacientes que concurrieron al laboratorio durante abril-diciembre 2019 y abril-diciembre 2021. Los pacientes COVID positivos fueron estratificados según el tiempo transcurrido desde la enfermedad en tres grupos: menos de 3 meses (fase aguda), 4-12 meses y más de 1 año. Resultados: Se halló diferencia significativa (p=0.013) en el recuento total de espermatozoides/eyaculado en el grupo COVID positivo con respecto al grupo prepandemia y COVID negativo (Mediana (Q1-Q3): 67.49 (26.42-139.44) vs. 102.48 (43.86-197.05) y 96.72 (38.22-189.27). Al estratificar el grupo COVID positivo según el tiempo transcurrido desde la enfermedad, la disminución significativa (p= 0.038) fue durante la fase aguda, recuperándose en el tiempo hasta llegar a valores similares a los grupos prepandemia y COVID negativo. El aislamiento social y la vacunación no tuvieron efectos significativos en las variables seminales. Discusión: El estudio evidenció cambios reversibles en la función testicular reflejado por la disminución de los espermatozoides en el total eyaculado de pacientes que habían tenido COVID-19. Estos cambios parecen estar relacionados con la fiebre y la inflamación y no a causa de la infección por el virus.
Assuntos
COVID-19 , Análise do Sêmen , Humanos , COVID-19/prevenção & controle , Masculino , Adulto , Pessoa de Meia-Idade , SARS-CoV-2 , Contagem de Espermatozoides , Pandemias , Sêmen/virologia , Fatores de Tempo , Motilidade dos Espermatozoides , Adulto JovemRESUMO
Viroids that belong to genera Avsunviroid and Pelamovirod (family Avsunviroidae) replicate and accumulate in the chloroplasts of infected cells. In this report, we confirmed by RNA in situ hybridization using digoxigenin-UTP-labelled riboprobes that the positive strands of eggplant latent viroid (ELVd), the only member of genus Elaviroid within the family Avsunviroidae, also accumulate in the chloroplasts of infected cells. However, comparison of ELVd in situ hybridization signals with those from bona fide chloroplastic and nuclear non-coding RNAs, such as chloroplast 5S rRNA and U1 small nuclear RNA, supports the notion that this viroid is also present in the nuclei of infected cells. These results suggest that the subcellular localization of viroids within the family Avsunviroidae may be more complex than previously assumed with dynamic presence in several compartments during the infectious cycle.
Assuntos
Núcleo Celular , Cloroplastos , Solanum melongena , Viroides , Viroides/genética , Viroides/fisiologia , Solanum melongena/virologia , Cloroplastos/virologia , Núcleo Celular/virologia , RNA Viral/genética , Hibridização In Situ , Doenças das Plantas/virologiaRESUMO
Eight porcine parvovirus (PPV) species, designated as PPV1 through PPV8, have been identified in swine. Despite their similarities, knowledge about their distribution and genetic differences remains limited, resulting in a gap in the genetic classification of these viruses. In this study, we conducted a comprehensive analysis using PPV1 to PPV7 genome sequences from Colombia and others available in the GenBank database to propose a classification scheme for all PPVs. Sera from 234 gilts aged 180 to 200 days were collected from 40 herds in Colombia. Individual detection of each PPV (PPV1 through PPV7) was performed using end-point PCR. Complete nucleotide (nt) sequencing was performed on the PPV1 viral protein (VP), and near-complete genome (NCG) sequencing was carried out for novel porcine parvoviruses (nPPVs) (PPV2 through PPV7). Phylogenetic analyses were conducted by comparing PPV1-VP sequences to 94 available sequences and nPPVs with 565 NCG, 846 nPPV-VP, and 667 nPPV-nonstructural protein (NS) sequences. Bayesian phylogenetic analysis was used to estimate substitution rates and the time to the most recent common ancestor for each PPV. The highest prevalence was detected for PPV3 (40.1%), followed by PPV5 (20.5%), PPV6 (17%), PPV1 (14.5%), PPV2 (9.8%), PPV4 (4.2%), and PPV7 (1.3%). Notably, all tested sera were negative for PPV8 genomes. An analysis of the PPV1-VP sequences revealed two main clades (PPV1-I and PPV1-II), with the sequences recovered in this study grouped in the PPV1-II clade. Comparative analysis showed significant genetic distances for PPV2 to PPV7 at the NCG (>6.5%), NS (>6.3%), and VP (>7.5%) regions, particularly when compared to equivalent regions of PPV genomes recovered worldwide. This study highlights the endemic circulation of nPPVs in Colombian pig herds, specifically among gilts. Additionally, it contributes to the phylogenetic classification and evolutionary studies of these viruses. The proposed method aims to categorize and divide subtypes based on current knowledge and the genomes available in databanks.
Assuntos
Genoma Viral , Infecções por Parvoviridae , Parvovirus Suíno , Filogenia , Doenças dos Suínos , Animais , Suínos , Parvovirus Suíno/genética , Parvovirus Suíno/classificação , Parvovirus Suíno/isolamento & purificação , Colômbia/epidemiologia , Doenças dos Suínos/virologia , Doenças dos Suínos/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Feminino , Epidemiologia Molecular , Evolução Molecular , Teorema de BayesRESUMO
The COVID-19 pandemic has overwhelmed healthcare systems and triggered global economic downturns. While vaccines have reduced the lethality rate of SARS-CoV-2 to 0.9% as of October 2024, the continuous evolution of variants remains a significant public health challenge. Next-generation medical therapies offer hope in addressing this threat, especially for immunocompromised individuals who experience prolonged infections and severe illnesses, contributing to viral evolution. These cases increase the risk of new variants emerging. This study explores miniACE2 decoys as a novel strategy to counteract SARS-CoV-2 variants. Using in silico design and molecular dynamics, blocking proteins (BPs) were developed with stronger binding affinity for the receptor-binding domain of multiple variants than naturally soluble human ACE2. The BPs were expressed in E. coli and tested in vitro, showing promising neutralizing effects. Notably, miniACE2 BP9 exhibited an average IC50 of 4.9 µg/mL across several variants, including the Wuhan strain, Mu, Omicron BA.1, and BA.2 This low IC50 demonstrates the potent neutralizing ability of BP9, indicating its efficacy at low concentrations.Based on these findings, BP9 has emerged as a promising therapeutic candidate for combating SARS-CoV-2 and its evolving variants, thereby positioning it as a potential emergency biopharmaceutical.
Assuntos
Enzima de Conversão de Angiotensina 2 , Anticorpos Neutralizantes , COVID-19 , Simulação de Dinâmica Molecular , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , SARS-CoV-2/efeitos dos fármacos , SARS-CoV-2/imunologia , Humanos , COVID-19/virologia , COVID-19/imunologia , Enzima de Conversão de Angiotensina 2/metabolismo , Enzima de Conversão de Angiotensina 2/química , Anticorpos Neutralizantes/imunologia , Glicoproteína da Espícula de Coronavírus/metabolismo , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/imunologia , Simulação por Computador , Pandemias , Ligação Proteica , Betacoronavirus/imunologia , Betacoronavirus/efeitos dos fármacos , Testes de NeutralizaçãoRESUMO
Bovine viral diarrhea virus (BVDV) causes ongoing economic losses to cattle industries, directly through reduced herd performance or indirectly through control program costs. ELISA assays, one of the most widely used techniques due to their ease of implementation, have been a valuable tool for mass surveillance and detection of BVDV. In this study, we developed a new indirect ELISA (rE2-ELISA) for serologic detection of BVDV. The assay considers three recombinant E2 protein subtypes as antigens, allowing serologic diagnosis of BVDV-1b (high prevalence worldwide), BVDV-1d and 1e (high prevalence in southern Chile) sub-genotypes. Recombinant E2 (rE2) proteins were successfully expressed in stably transfected CHO cells. Conditions for rE2 ELISAs were established after determining appropriate concentrations of antigen, blocking agent, secondary antibody, and serum dilutions to achieve maximum discrimination between positive and negative serum samples. The developed rE2-ELISA showed a sensitivity of 92.86% and a specificity of 98.33%. Clinical testing of 180 serum samples from herds in southern Chile showed high accuracy (kappa > 0.8) compared to the commercial BVDV Total Ab kit (IDEXX), with 95.37% positive and 87.5% negative predictive value. In addition, the rE2 ELISA has shown the capability to detect anti-BVDV antibodies from naturally infected animals with sub-genotypes 1b, 1e, or undetermined. These results indicate that the developed indirect ELISA could serve as a valid, and efficient alternative for identifying BVDV-infected animals, thus contributing to the success of disease control and eradication programs.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Bovinos , Doença das Mucosas por Vírus da Diarreia Viral Bovina/diagnóstico , Doença das Mucosas por Vírus da Diarreia Viral Bovina/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Chile , Genótipo , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina/imunologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Proteínas do Envelope Viral/imunologia , Proteínas do Envelope Viral/genética , Antígenos Virais/imunologia , Cricetulus , Células CHO , Anticorpos Antivirais/sangue , Proteínas Recombinantes/imunologiaRESUMO
Panama is a country with endemic Dengue virus (DENV) transmission since its reintroduction in 1993. The four serotypes have circulated in the country and the region of the Americas, however, DENV-4 confirmed autochthonous cases have not been identified since 2000, despite its circulation in neighboring countries. Here, we report DENV-4 detection in Panama in the last four-month period of 2023 with co-circulation of the other serotypes, this was associated with a peak of dengue cases during the dry season even though most dengue outbreaks are described in the rainy season. Complete genomes of DENV-4 allowed us to determine that cases were caused by DENV-4 genotype IIb, the same genotype as 23 years ago, with high similarity to DENV-4 sequences circulating in Nicaragua and El Salvador during 2023. This report shows the importance of maintaining serotype and genotype surveillance for early detection of new variants circulating in the country.
Assuntos
Vírus da Dengue , Dengue , Genoma Viral , Genótipo , Filogenia , Sorogrupo , Vírus da Dengue/genética , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , Panamá/epidemiologia , Dengue/epidemiologia , Dengue/virologia , Humanos , Genoma Viral/genética , RNA Viral/genética , Estações do Ano , Surtos de Doenças , Nicarágua/epidemiologiaRESUMO
It is heavily suggested that one IFNL4 gene polymorphism, rs12979860 (T/C), exerts influence on the outcome of HBV infection, with the rs12979860-T allele being classified as a risk predictor, and the rs12979860-C allele being classified as a protective one. This study investigated whether the rs12979860 IFNL4 gene polymorphism presented any association with the clinical severity for HBV carriers in an admixed population in Northern Brazil. A total of 69 samples were investigated from infected people from the city of Belém-Pará. The rs12979860-T allele was positively associated with HBV infection, suggesting a higher risk of chronicity. This research's importance is that the polymorphism influence was investigated in a population of HBV carriers with a heterogeneous genetic profile, formed through the extensive admixture of different ethnic groups, including Europeans, Africans, and Natives with indigenous heritage. This analysis is particularly important since highly mixed populations do not always follow the same association patterns previously established by studies using populations classified as more genetically homogeneous, due to a different formation process.
Assuntos
Predisposição Genética para Doença , Vírus da Hepatite B , Hepatite B , Interleucinas , Polimorfismo de Nucleotídeo Único , Humanos , Brasil/epidemiologia , Masculino , Interleucinas/genética , Feminino , Adulto , Vírus da Hepatite B/genética , Hepatite B/genética , Hepatite B/virologia , Pessoa de Meia-Idade , Alelos , Frequência do Gene , Genótipo , Interferon lambdaRESUMO
Conventional live virus research on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of coronavirus disease-19 (COVID-19), requires Biosafety Level 3 (BSL-3) facilities. SARS-CoV-2 pseudotyped viruses have emerged as valuable tools in virology, mimicking the entry process of the SARS-CoV-2 virus into human cells by expressing its spike glycoprotein in a surrogate system using recombinant plasmids. One significant application of this tool is in functional assays for the evaluation of neutralizing antibodies. Pseudotyped viruses have the advantage of being competent for only a single cycle of infection, providing better safety and versatility and allowing them to be studied in BSL-2 laboratories. Here, we describe three protocols for the detection of SARS-CoV-2 neutralizing antibodies through a pseudotyped virus assay. First, SARS-CoV-2 S pseudotyped viruses (PV SARS-CoV-2 S) are produced using a Moloney murine leukemia virus (MuLV) three-plasmid system. The plasmids are designed to express the GagPol packing proteins, enhanced green fluorescent protein (eGFP) as a readout system, and the SARS-CoV-2 S protein modified to remove the endoplasmic reticulum retention domain and to improve infection. Next, the internalization of PV SARS-CoV-2 S protein in human embryonic kidney 293T (HEK-293T) cells overexpressing angiotensin-converting enzyme 2 (HEK-293T-ACE2) is confirmed by fluorescence microscopy and quantified using flow cytometry. Finally, PV SARS-CoV-2 S is used to screen neutralizing antibodies in serum samples from convalescent COVID-19 patients; it can also be used for studying the cell entry mechanisms of different SARS-CoV-2 variants, evaluating antiviral agents, and designing vaccines. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Generation of PV SARS-CoV-2 S pseudotyped virus Basic Protocol 2: Assay of PV SARS-CoV-2 S internalization in target cells. Basic Protocol 3: Detection of neutralizing antibodies in serum samples.
Assuntos
Anticorpos Neutralizantes , Anticorpos Antivirais , COVID-19 , SARS-CoV-2 , Humanos , Anticorpos Neutralizantes/imunologia , Anticorpos Neutralizantes/sangue , SARS-CoV-2/imunologia , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , COVID-19/virologia , COVID-19/imunologia , COVID-19/diagnóstico , COVID-19/sangue , Testes de Neutralização/métodos , Células HEK293 , Pseudotipagem Viral , Glicoproteína da Espícula de Coronavírus/imunologia , Glicoproteína da Espícula de Coronavírus/genéticaRESUMO
Feline calicivirus (FCV) icosahedral viral capsids are composed of dozens of structural subunits that rely on cellular chaperones to self-assemble in an orderly fashion. Here, we report that the heat shock protein 90 (Hsp90) inhibition significantly reduced FCV particle production, suggesting a role in the replicative cycle. We found that Hsp90 inhibition was not related to the synthesis or stability of the early proteins that translate from the gRNA nor to the minor capsid protein VP2 but with a reduction in the major capsid protein VP1 levels, both translated late in infection from the subgenomic RNAs. Reduction in VP1 levels was observed despite an augment of the leader of the capsid (LC)-VP1 precursor levels, from which the LC and VP1 proteins are produced after proteolytic processing by NS6/7. The direct interaction of VP1 with Hsp90 was observed in infected cells. These results suggest that upon release from the polyprotein precursor, VP1 becomes a client of Hsp90 and that this interaction is required for an efficient FCV replicative cycle.
Assuntos
Calicivirus Felino , Proteínas do Capsídeo , Proteínas de Choque Térmico HSP90 , Replicação Viral , Proteínas de Choque Térmico HSP90/metabolismo , Proteínas de Choque Térmico HSP90/genética , Proteínas do Capsídeo/metabolismo , Proteínas do Capsídeo/genética , Calicivirus Felino/metabolismo , Calicivirus Felino/fisiologia , Calicivirus Felino/genética , Gatos , Animais , Linhagem Celular , Infecções por Caliciviridae/virologia , Infecções por Caliciviridae/metabolismoRESUMO
Benign childhood myositis is a self-limiting inflammatory condition that primarily affects schoolaged boys during the winter months. It is associated with respiratory viral infections, such as influenza A and B viruses, respiratory syncytial virus (RSV), and Mycoplasma pneumoniae, among others. In September 2022, an epidemiological alert was raised due to a high number of reported cases in the metropolitan area of Buenos Aires. We present two cases of female adolescents, aged 10 and 14 years, who developed this condition in association with influenza B virus infection. Their treatment and follow-up differed based on their clinical presentation and laboratory findings. This entity should be considered in the differential diagnosis of lower limb myalgia, difficulty walking, and functional impotence. It is necessary to establish management guidelines according to the clinic and laboratory. The search for respiratory viruses, mainly influenza, should be done taking into account the local epidemiology.
La miositis benigna de la infancia es un cuadro inflamatorio, asociado a infecciones virales respiratorias, autolimitado que no deja secuelas. Afecta principalmente a varones en edad escolar, durante los meses de invierno. Puede asociarse a virus influenza A y B, VRS, Mycoplasma pneumoniae, entre otros. En el mes de septiembre de 2022 se generó una alerta epidemiológica por el elevado número de casos reportados en el área metropolitana de Buenos Aires. Presentamos los casos de dos adolescentes mujeres de 10 y 14 años con este cuadro asociado a influenza B. El tratamiento y el seguimiento fueron diferentes en base a la clínica y el laboratorio. Hay que tener en cuenta esta entidad como un diagnóstico diferencial de mialgias de miembros inferiores, dificultad en la marcha e impotencia funcional. Es necesario establecer las pautas de manejo según clínica y laboratorio. La búsqueda de virus respiratorios, principalmente influenza, debe hacerse teniendo en cuenta la epidemiología local.