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Arboviruses cause public health problems in several countries, and records show that they can generate central and peripheral neurological complications with permanent sequelae. However, it is not certain which arbovirus is responsible for outbreaks of the Guillain-Barré Syndrome (GBS), especially in Brazil. Thus, the objective of this study is to verify if there is a coincidence between the GBS outbreak and the most common arboviruses in Northeastern Brazil, as well as their relationship. An ecological time series study was designed with the federative units of Northeastern Brazil, using hospitalizations for Guillain-Barré syndrome and notifications of arbovirus infections between 2014 and 2019 as a data source. Distribution incidence curves were constructed for the conditions studied, and generalized estimating equations (GEE) models were applied to estimate the relationship between arboviruses and Guillain-Barré. The results showed a similar distribution for the incidences of Chikungunya virus (z=7.82; p=0.001), Zika virus (z=3.69; p=0.03), and Guillain-Barré syndrome (z=2.98; p=0.05) from 2014 to 2019. The GEE model revealed that the distribution of Chikungunya incidence is associated with the distribution of GBS incidence in each year (x2Wald=3,969; p=0.046). This pattern was repeated in seven of the nine states, while the Zika virus had a significant relationship with GBS in only two states. The outbreak of GBS in Northeastern Brazil appears to be probabilistically related to outbreaks of the Chikungunya virus.
As arboviroses são problemas de saúde pública em vários países e há registros de que podem produzir complicações neurológicas centrais e periféricas com sequelas permanentes. Entretanto, não se sabe ao certo qual delas é realmente responsável pelos surtos da Síndrome de Guillain-Barré (SGB), principalmente no Brasil. Assim, o objetivo é verificar se há coincidência entre o surto de SGB e as arboviroses mais comuns no Nordeste do Brasil e suas relações. Foi desenhado um estudo ecológico de série temporal com as unidades federativas do Nordeste do Brasil, adotando como fonte de dados as internações Guillain-Barré e as notificações de infecções por arbovírus entre 2014 e 2019. Curvas de distribuição de incidência foram construídas para as condições estudadas, e foram aplicados modelos de equações generalizadas estimadas (GEE) para estimar a relação entre arbovírus e Guillain-Barré. Evidencia-se que há distribuição semelhante para as incidências do vírus Chikungunya (z=7,82; p=0,001), vírus Zika (z=3,69; p=0,03) e síndrome de Guillain-Barré (z=2,98; p=0,05) entre 2014 e 2019. O modelo GEE revelou que a distribuição da incidência de Chikungunya está associada à distribuição da incidência de SGB em cada ano (x2Wald=3,969; p=0,046). Esse padrão se repetiu em sete dos nove estados, enquanto o zika vírus teve uma relação significativa com o GBS em apenas dois estados. Conclui-se, então, que o surto de SGB no Nordeste do Brasil parece estar probabilisticamente relacionado aos surtos do vírus Chikungunya.
Los arbovirus causan problemas de salud pública en varios países y, según indican los reportes, pueden producir complicaciones neurológicas centrales y periféricas con secuelas permanentes. Sin embargo, no se sabe cuál de ellos es realmente el responsable de los brotes del síndrome de Guillain-Barré (SGB), especialmente en Brasil. Así, el objetivo de este estudio es verificar si existen coincidencias entre el brote del SGB y los arbovirus más comunes en el Noreste de Brasil y sus asociaciones. Se diseñó un estudio de series temporales ecológico en las unidades federativas del Noreste de Brasil, adoptando como fuente de datos las hospitalizaciones y las notificaciones de arbovirosis de Guillain-Barré entre 2014 y 2019. Se construyeron curvas de distribución de incidencia para las condiciones científicas, y se aplicó una ecuación estimada generalizada (GEE) para estimar la relación entre arbovirus y Guillain-Barré. Se encontró que existe una distribución similar en las incidencias de virus del chikunguña (z=7,82; p=0,001), virus del Zika (z=3,69; p=0,03) y síndrome de Guillain-Barré (z =2,98; p=0,05) entre 2014 y 2019. El modelo GEE reveló que la distribución de la incidencia de chikunguña está asociada con la distribución de la incidencia de SGB en cada año (x2Wald=3,969; p=0,046). Este patrón se repitió en siete de los nueve estados, mientras que el virus del Zika presentó una relación significativa con el SGB en solo dos estados. El brote del SGB en el Noreste de Brasil parece estar relacionado probabilísticamente con los brotes del virus del chikunguña.
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ABSTRACT We conducted a spatial case-control study nested in a dengue incidence cohort to explore the role of the spatial and socioeconomic factors in the proportion of symptomatic (cases) and inapparent primary dengue virus infections (controls). Cohort participants were children and adolescents (2 to 16 years of age) at the beginning of the follow-up. Case definitions were, for symptomatic cases, fever plus a positive lab result for acute dengue (NS1, RT-PCR, ELISA IgM/IgG), and for inapparent infection a positive result for dengue IgG (ELISA) in subjects without symptoms and with a previously negative result at baseline. The covariates included sociodemographic factors, residential location, and socioeconomic context variables of the census tracts of residence of cases and controls. We used principal component analysis to reduce the contextual covariates, with the component values assigned to each one based on their residences. The data were modeled in a Bayesian context, considering the spatial dependence. The final sample consisted of 692 children, 274 cases and 418 controls, from the first year of follow-up (2014-2015). Being male, older age, higher educational level of the head of the family and having a larger number of rooms in the household were associated with a greater chance of presenting dengue symptomatic infection at the individual level. The contextual covariates were not associated with the outcome. Inapparent dengue infection has extensive epidemiological consequences. Relying solely on notifications of symptomatic dengue infections underestimates the number of cases, preserves a silent source of the disease, potentially spreading the virus to unaffected areas.
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Resumo O estabelecimento definitivo do Aedes aegypti nas Américas está associado às mudanças ambientais, urbanização desorganizada, ausência de água e saneamento básico e deslocamentos populacionais, definindo, assim, a trajetória da doença. O objetivo deste estudo foi analisar a distribuição espacial dos casos suspeitos de dengue e a relação com indicadores socioeconômicos no município de São Carlos-SP. Trata-se de estudo ecológico, realizado nesse município, em que foram utilizados dados secundários do Sistema de Informação sobre Agravos de Notificação no período de 2016 a 2017 e do Instituto Brasileiro de Geografia e Estatística do Censo Demográfico de 2010. Os dados foram analisados por meio da Análise de Componentes Principais pelo software Statistica 12.0 e os mapas temáticos foram gerados pelo software Arcgis versão 10.5. Os resultados mostraram elevadas taxas de casos suspeitos de dengue em áreas de ponderação com condições sociais adequadas e taxas reduzidas em áreas de ponderação com iniquidades sociais. Concluiu-se que o perfil de casos suspeitos por dengue na população de São Carlos não está relacionado a baixos indicadores socioeconômicos, uma vez que a dinâmica e a mobilidade populacional relacionadas ao elevado fluxo de universitários podem influenciar no padrão de distribuição espacial da doença.
Abstract The definitive establishment of Aedes aegypti in the Americas is associated with environmental changes, disorganized urbanization, lack of water and basic sanitation and population displacement, thus defining the trajectory of the disease. The aim of this study was to analyze the spatial distribution of the dengue suspected cases and understand its correlation with the socioeconomic indicators in the city of São Carlos (SP). This is an ecological study, carried out at the São Carlos (SP). Secondary data from the Information System on Notifiable Diseases in the period from 2016 to 2017 and from the Brazilian Institute of Geography and Statistics of the 2010 Demographic Census were used. The data were analyzed using Principal Component Analysis (ACP) by the software Statistica 12.0 and the thematic maps were generated by the software Arcgis version 10.5. The results showed high rates of suspected dengue cases in areas with adequate social conditions and reduced rates in areas with worst social condition. It was concluded that the outline of suspected dengue cases in the population of São Carlos (SP) is not related to low socioeconomic indicators, once the population dynamics and mobility related to the high flow of university students can influence the pattern of spatial distribution of the disease.
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Cervical artery dissections (CAD) can occur spontaneously or as a direct result of significant trauma. Viral infections, such as SARS-CoV2, influenza, and Epstein Barr, are risk factors for spontaneous CAD. Dengue virus infections have dramatically increased in recent decades, and Brazil is one of the endemic areas. The dengue virus can cause headache and neurological complications such as encephalitis, myelitis, Guillain-Barré syndrome, and myositis. No report has yet been found in the literature of dissection of the internal carotid artery secondary to dengue infection. Our objective is to report the case of a patient with dissection of the internal carotid artery associated with acute dengue virus infection.
As dissecções da artéria cervical (DAC) podem ocorrer espontaneamente ou como resultado direto de trauma significativo. Infecções virais, como SARS-CoV2, influenza e Epstein Barr, são fatores de risco para DAC espontânea. As infecções pelo vírus da dengue aumentaram dramaticamente nas últimas décadas, e o Brasil é uma das áreas endêmicas. O vírus da dengue pode causar dor de cabeça e complicações neurológicas como encefalite, mielite, síndrome de Guillain-Barré e miosite. Ainda não foi encontrado na literatura nenhum relato de dissecção da artéria carótida interna secundária à infecção por dengue. Nosso objetivo é relatar o caso de um paciente com dissecção da artéria carótida interna associada à infecção aguda pelo vírus da dengue.
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Humans , Virus Diseases/epidemiology , Dengue Virus/immunology , Carotid Artery, Internal, Dissection/classification , Dengue/diagnosis , Dissection/methodsРеферат
Abstract Immunodiagnostic tests for detecting dengue virus infections encounter challenges related to cross-reactivity with other related flaviviruses. Our research focuses on the development of a synthetic multiepitope antigen tailored for dengue immunodiagnostics. Selected dengue epitopes involved structural linearity and dissimilarity from the proteomes of Zika and Yellow fever viruses which served for computationally modeling the three-dimensional protein structure, resulting in the design of two proteins: rDME-C and rDME-BR. Both proteins consist of seven epitopes, separated by the GPGPG linker, and a carboxy-terminal 6 × -histidine tag. The molecular weights of the final proteins rDME-C and rDME-BR are 16.83 kDa and 16.80 kDa, respectively, both with an isoelectric point of 6.35. The distinguishing factor between the two proteins lies in the origin of their epitope sequences, where rDME-C is based on the reference dengue proteome, while rDME-BR utilizes sequences from prevalent Dengue genotypes in Brazil from 2008 to 2019. PyMol analysis revealed exposure of epitopes in the secondary structure. Successful expression of the antigens was achieved in soluble form and fluorescence experiments indicated a disordered structure. In subsequent testing, rDME-BR and rDME-C antigens were assessed using an indirect Elisa protocol against Dengue infected serum, previously examined with a commercial diagnostic test. Optimal concentrations for antigens were determined at 10 µg/mL for rDME-BR and 30 µg/mL for rDME-C, with serum dilutions ranging from 1:50 to 1:100. Both antigens effectively detected IgM and IgG antibodies in Dengue fever patients, with rDME-BR exhibiting higher sensitivity. Our in-house test showed a sensitivity of 77.3 % and 82.6 % and a specificity of 89.4 % and 71.4 % for rDME-C and rDEM-BR antigens. No cross-reactivity was observed with serum from Zika-infected mice but with COVID-19 serum samples. Our findings underscore the utility of synthetic biology in crafting Dengue-specific multiepitope proteins and hold promise for precise clinical diagnosis and monitoring responses to emerging Dengue vaccines.
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ABSTRACT Objective: To determine the prevalence of ocular alterations in patients diagnosed with dengue, admitted to a reference center for infectious and parasitic diseases in Manaus (AM), Brazil. Methods: This was an observational, analytical, and prospective case series study with 33 patients admitted to the hospital with positive clinical pathology tests during a 90-day period of a dengue epidemic in 2011. The investigations included measurement of visual acuity, macular assessment with Amsler grid test, fundus biomicroscopy, retinography and optical coherence tomography. Results: The average age of the patients was 35.48 years (minimum age 17 years and maximum 69 years). The main symptoms reported were blurred vision, followed by retro-ocular pain, photopsia, halos, foreign bodies, scotoma, double vision, and floaters. Visual acuity ranged from 20/20 to 20/40 (median 20/30). Macular assessment with Amsler Grid Test was unsatisfactory in three patients. The most common retinal findings were perimacular edema, macular edema, retinal hemorrhage, vasculitis, and changes in the retinal pigment epithelium. Optical coherence tomography showed retinal edema in 16 patients (51.51%). Conclusions: In all cases, the disease was self-limiting and resolved spontaneously without treatment. Therefore, this study confirms that ocular manifestations in dengue fever are varied and frequent.
RESUMO Objetivo: Determinar a prevalência de alterações oculares em pacientes com diagnóstico de dengue, internados em um centro de referência para doenças infecciosas e parasitárias em Manaus (AM), Brasil. Métodos: Estudo observacional, analítico e prospectivo de série de casos de 33 pacientes admitidos no hospital com exames de patologia clínica positivos durante um período de 90 dias de uma epidemia de dengue em 2011. As investigações incluíram medida da acuidade visual, avaliação macular com tela de Amsler, biomicroscopia de fundo de olho, retinografia e tomografia de coerência ótica. Resultados: A idade média dos pacientes foi de 35,48 anos (idade mínima de 17 anos e máxima de 69 anos). Os principais sintomas relatados foram visão turva, seguida de dor retro-ocular, fotopsia, halos, corpos estranhos, escotoma, visão dupla e moscas volantes. A acuidade visual variou entre 20/20 e 20/40 (mediana 20/30). A avaliação macular com a tela de Amsler foi insatisfatória em três doentes. Os achados retinianos mais comuns foram edema perimacular, edema macular, hemorragia retiniana, vasculite e alterações no epitélio pigmentar da retina. A tomografia de coerência óptica mostrou edema da retina em 16 doentes (51,51%). Conclusões: Em todos os casos, a doença foi autolimitada e resolveu-se espontaneamente sem tratamento. Este estudo confirma, portanto, que as manifestações oculares da dengue são variadas e frequentes.
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The lack of effective conventional therapie s against dengue has created an interest in herbal preparations as alternative therapies. In the present study, in vitro effects of Cordia curassavica essential oil (EO) on both dengue virus replication and cytokine production were examined. Predictions of molecular interactions between EO compounds and virus and cell proteins were performed with AutoDock Vina. The EO inhibited replication of dengue virus serotypes at IC 50 < 30 µg/mL, and it reduced 87% TNF - α, 67% IL - 8 and 46% IFN - α in LPS - stimulated PBMCs. The main EO compounds were trans - ß - caryophyllene (21.4%), germacrene D (17.8%), α - copaene (16.5%), trans - ß - guaiene (8.2%), and α - pinene (6.0%). The first two compounds, δ - cadinene, α - muurolene, α - cubebene and ß - burbonene were coupled to proteins involved in the TLR - 4 cytokine effector pathway. 3,7 - Guaiadiene was coupled to the viral E and C proteins. This study demonstrates the potential of C. curassavica EO as a starting point for discovering novel therapeutic for dengue.
La falta de terapias eficaces para el dengue ha suscitado interés por preparados herbales como terapias alternativas. En el presente estudio se examinaron efectos in vitro del aceite e sencial (AE) de Cordia curassavica sobre la replicación del virus dengue y producción de citoquinas. Se realizaron predicciones de interacciones moleculares entre los compuestos del AE y proteínas virales y celulares con AutoDock Vina. El AE inhibió la rep licación de serotipos del virus a CI 50 < 30 µg/mL y redujo 87% TNF - α, 67% IL - 8 y 46% IFN - α en MNCP. Los principales compuestos del AE fueron trans - ß - cariofileno, germacreno D, α - copaeno, trans - ß - guaieno y α - pineno. Los dos primeros compuestos, el δ - cadineno, el α - muuroleno, el α - cubebeno y el ß - burboneno se acoplaron a proteínas implicadas en la vía efectora de citoquinas TLR - 4. El 3,7 - guaiadiene se acopló a las proteínas virales E y C. Este estudio demuestra el potencial del AE de C. curassavica como punto de partida para descubrir nuevas tera pias para el dengue.
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Oils, Volatile/pharmacology , Cordia/chemistry , Dengue/drug therapy , Plants, Medicinal/chemistry , Plant Oils/pharmacology , Cytokines/chemistryРеферат
INTRODUCCIÓN. El dengue es una enfermedad infecciosa de origen viral, transmitida principalmente por el mosquito Aedes aegypti. Es un grave problema de salud pública a nivel mundial, en las Américas y en el Ecuador. OBJETIVOS. Analizar el comportamiento epidemiológico del dengue desde 1980 hasta el 2020, los factores de riesgo que mantienen la transmisión y las acciones que el país ha implementado para su prevención y control. METODOLOGÍA. Revisión bibliográfica narrativa, teniendo como fuente las páginas web de la Organización Mundial de la Salud, de la Organización Panamericana de la Salud, Ministerio de Salud Pública del Ecuador, artículos de revistas de bibliotecas virtuales e informes técnicos publicados en Google académico, Scielo, Biblioteca Virtual en Salud y PubMed. RESULTADOS. Los registros encontrados sobre presencia de casos de dengue datan de 1988 cuando se presentó la gran epidemia de dengue en Guayaquil. Posteriormente, mantiene un comportamiento endemo-epidémico. A partir del año 2000 se presentan casos de dengue grave. Su pico más alto de letalidad fue de 2,44% en el 2010. El serotipo DEN 1 es el más frecuente, pero a partir del 2000 circulan los 4 serotipos. Las acciones de prevención y control no han sido sostenidas. CONCLUSIONES. El dengue en Ecuador en los últimos cuarenta años mantiene una importante trasmisión, caracterizada por años epidémicos. No ha podido ser controlado el vector, el cual ha ido infestando más localidades. Las condicionantes climáticas y ecológicas, explican en parte la persistencia, pero la determinación más importante está dada por las inequidades sociales, falta de servicios básicos, y la poca continuidad e impacto de las medidas de prevención y control.
INTRODUCTION. Dengue is an infectious disease of viral origin, transmitted mainly by the Aedes aegypti mosquito. It is a serious public health problem worldwide, in the Americas and in Ecuador. OBJECTIVES: To analyze the epidemiological behavior of dengue fever from 1980 to 2020, the risk factors that maintain transmission and the actions that the country has implemented for its prevention and control. METHODOLOGY. Narrative bibliographic review, taking as sources the web pages of the World Health Organization, Pan American Health Organization, Ministry of Public Health of Ecuador, journal articles from virtual libraries and technical reports published in Google Scholar, Scielo, Virtual Health Library and PubMed. RESULTS. The records found on the presence of dengue cases date back to 1988 when the great dengue epidemic occurred in Guayaquil. Subsequently, it maintained an endemic-epidemic behavior. Beginning in 2000, severe cases of dengue fever occurred. Its highest lethality peak was 2.44% in 2010. DEN 1 serotype is the most frequent, but since 2000 all 4 serotypes have been circulating. Prevention and control actions have not been sustained. CONCLUSIONS. Dengue in Ecuador over the last forty years has maintained an important transmission, characterized by epidemic years. It has not been possible to control the vector, which has been infesting more localities. Climatic and ecological conditions partly explain its persistence, but the most important determinant is given by social inequalities, lack of basic services, and the lack of continuity and impact of prevention and control measures.
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Humans , Male , Female , Risk Factors , Aedes , Dengue , Dengue Virus , Ecuador , Insect Vectors , Epidemiology , Severe DengueРеферат
Introducción: El dengue se ha convertido en un problema serio de salud pública en Bolivia. Por la capacidad de infección del virus del dengue, el diagnóstico es amplio y varia con la evolución de la enfermedad. El incremento de la temperatura y las lluvias de la época son los factores ambientales principales para la reproducción del mosquito transmisor (Aedes aegypti), y el aumento de casos de dengue. Objetivos: Realizar un estudio transversal-retrospectivo de los casos de dengue en el departamento de La Paz - Bolivia, durante el periodo de enero de 2020 hasta febrero de 2023. Materiales y Métodos: De las bases obtenidas del Laboratorio de Virología- INLASA, se analizaron variables como el número de casos positivos de cada gestión, la distribución de los casos positivos según el lugar probable de infección tanto por departamento como por provincia, la variación temporal de los casos y el serotipo prevalente por gestión, y la distribución de casos positivos por género y grupo etario, en este trabajo se realizó un estudio transversal-retrospectivo. Resultados: Durante las cuatro gestiones analizadas el número total de casos notificados fue de 2143, detectando 1184 (55.2%) positivos. Señalando como lugar probable de infección en gran parte la provincia de Sud Yungas. Con respecto a la variación temporal presentó un mismo comportamiento en tres gestiones. Prevaleció en tres gestiones el serotipo DENV 1 sobre DENV 2, observando un aumento de serotipo DENV 2 hasta febrero del 2023. Además, el grupo atareo más afectado con dengue oscila entre los 10 a 39 años, no encontrando diferencia significativa entre género masculino y femenino. Conclusiones: Este estudio revela que el dengue es un problema de salud pública no solo del oriente del país sino también de las zonas tropicales del departamento de La Paz. Es necesario el diagnóstico rápido y efectivo; realizar la tipificación del virus y el llenado correcto de fichas epidemiológicas para un mejor control virológico.
Background: Dengue has become a serious problem in public health in Bolivia. Due the infection capacity of dengue virus, the diagnosis is broad and varies with the evolution of the disease. The increase of temperature and rain season are the main environmental factors of reproduction in the transmitting mosquito (Aedes aegipty), and the increase in dengue cases. Objectives: To carry out a retrospective crossover study of dengue cases in La Paz Department - Bolivia, during the period from January 2020 to February 2023. Materials and Methods: From databases obtained from INLASA Virology Laboratory during periods 2020, 2021, 2022 and the beginnings of 2023, variables such as number of positive cases from each year, distribution of positive cases according to the probable place of infection were obtained and analyzed, by department and by province; the temporal variation of the cases and the prevalent serotype by period, and finally distribution of positive cases by sex group and age group. Its make a retrospective crossover study. Results: During the four analyzed periods, within the total number of 2143 reported cases, there were 1184 (55.2%) of positive cases for Dengue. On the other hand, the department of La Paz (86.3%) is indicated as the probable place of infection, with Sud Yungas province (municipalities La Asunta and Palos Blancos) being the places with the most positive cases. The temporal variation in suspected and confirmed cases has presented the same behavior in three years. DENV 1 serotype predominated over DENV 2 in three years, observing an increase in DENV 2 serotype until February 2023. In addition, the age group most affected by dengue ranges from 10 to 39 years, finding no significant difference between masculine and feminine gender. Discussions: An increase in cases between months of February and April, coinciding with rain season in Bolivia. Due to the climate in the tropical areas of the department, there is a greater number of positive cases in those areas. Conclusions: This study reveals that Dengue is a public health problem not only in eastern part of the country but also in tropical areas of La Paz department. A quick and effective diagnosis is necessary; to carry out the typing of the virus and the correct filling out of epidemiological files for better virological control.
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Spontaneous mutations and lack of replication fidelity in positive-sense single stranded RNA viruses (+ssRNA virus) result in emergence of genetic variants with diverse viral morphogenesis and surface proteins that affect its antigenicity. This high mutability in +ssRNA viruses has induced antiviral drug resistance and ability to overcome vaccines that subsequently resulted in rapid viral evolution and high mortality rate in human and livestock. Computer aided vaccine design and immunoinformatics play a crucial role in expediting the vaccine production protocols, antibody production and identifying suitable immunogenic regions or epitopes from the genome sequences of the pathogens. T cell and B cell epitopes can be identified in pathogens by immunoinformatics algorithms and methods that enhance the analysis of protective immunity, vaccine safety, immunity modelling and vaccine efficacy. This rapid and cost-effective computational vaccine design promotes development of potential vaccine that could induce immune response in host against rapidly mutating pathogens like +ssRNA viruses. Epitope-based vaccine is a striking concept that has been widely employed in recent years to construct vaccines targeting rapidly mutating +ssRNA viruses. Therefore, the present review provides an overview about the current progress and methodology in computeraided vaccine design for the most notable +ssRNA viruses namely Hepatitis C virus, Dengue virus, Chikungunya virus and Coronaviruses. This review also highlights the applications of various immunoinformatics tools for vaccine design and for modelling immune response against +ssRNA viruses.
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@#ObjectiveTo establish models of Dengue virus type Ⅲ(DENV-3,DV-3)infection and antibody dependent enhancement(ADE)infection at the acute monocytic leukemia cells(THP-1),investigate the differential expression of long non-coding RNAs(LncRNAs),map the competitive endogenous RNA(CeRNA)regulatory network and predict the translation function of LncRNAs.MethodsThe culture supernatant was harvested 6 d after C6/36 cells were infected with DENV-3,the virus titer was determined by CCID50,and the type and full-length genome amplification were identified by PCR;The DENV-3 standard plasmid was amplified,identified by PCR,and the standard curve was drawn;THP-1 cells were divided into negative control group(THP-1),direct infection group(DV-3),ADE group and blank control group[1640(-)]. After 48 h of infection,the total RNA was extracted and the copy number of intracellular virus nucleic acid was measured;Through the whole transcriptome sequencing technology,the CeRNA regulatory network was constructed for the top five up-regulated and down-regulated LncRNAs in THP-1 vs DENV3,THP-1 vs ADE,DENV3 vs ADE groups,and the functions of their coding proteins were analyzed.ResultsC6/36 cells infected with DENV-3 for 3 d showed obvious cell fusion,vacuoles and abscission;The virus had a titer of about 1. 0 × 104. 64PFU/mL and was identified as DENV-3 by PCR specific primers,of which the complete gene sequence was obtained;The number of viral nucleic acid copies in ADE group was significantly higher than those in DV-3 group and blank control group;In THP-1 vs DENV-3,the expression of cytohesin interacting protein(CYTIP)was predicted to be up-regulated;In THP-1 vs ADE,the expression of kinesin family5A(KIF5A)was predicted to be down-regulated;In DENV-3 vs ADE,the expression of cluster differentiation antigen 9(CD9)and insulin like growth factor 2(IGF2)was predicted to be up-regulated. All of these differential LncRNAs had open reading frames(ORFs). Except Lnc-SH3BP1 and Lnc-RPL41,all of the other LncRNAs had internal ribosome binding site(IRES).ConclusionIn DENV-3 infection of THP-1 cells and ADE infection mediated by DENV-3,the expression of LncRNAs has changed significantly,and may regulate the process of infection through a variety of biological functions,which is helpful for a deeper understanding of the mechanism of ADE infection.
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Aims@#Dengue virus is a global pathogen that lacks an effective vaccine or therapy. Screening medicinal plants for anti-dengue properties provides a promising avenue to identify potent compounds. Mangroves, known for their resilience in harsh conditions, produce a diverse range of natural products with unique biochemical profiles, which hold potential for anti-dengue treatments. This study aims to evaluate the anti-dengue activity of selected mangrove plant species from Sabah against DENV2 NS2B-NS3pro, utilizing an enzymatic protease assay. @*Methodology and results @#Six mangrove species (Avicennia marina, Bruguiera gymnorrhiza, Ceriops tagal, Rhizophora apiculata, Rhizophora mucronata and Xylocarpus granatum) were investigated, with various plant parts subjected to aqueous and ethanol extraction. The results demonstrated significant anti-dengue activity in both aqueous and ethanolic extracts of the mangroves against DENV2 NS2B-NS3pro, with IC50 values ranging from 0.95 µg/mL to 6.24 µg/mL. Notably, the ethanolic extract of R. apiculata leaves exhibited the highest inhibition, with an IC50 value of 0.95 µg/mL. @*Conclusion, significance and impact of study@#These findings suggest that the ethanolic extracts from R. apiculata leaves hold promise as potential candidates for dengue treatment. This study underscores the importance of natural products as valuable sources for the development of novel anti-dengue treatments, highlighting the need to explore mangroves in the quest for effective therapeutic options.
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@#Bats are flying mammals with unique immune systems that allow them to hold many pathogens. Hence, they are recognised as the reservoir of many zoonotic pathogens. In this study, we performed molecular detection to detect coronaviruses, paramyxoviruses, pteropine orthoreoviruses and dengue viruses from samples collected from insectivorous bats in Krau Reserve Forest. One faecal sample from Rhinolophus spp. was detected positive for coronavirus. Based on BLASTN, phylogenetic analysis and pairwise alignment-based sequence identity calculation, the detected bat coronavirus is most likely to be a bat betacoronavirus lineage slightly different from coronavirus from China, Philippines, Thailand and Luxembourg. In summary, continuous surveillance of bat virome should be encouraged, as Krau Reserve Forest reported a wide spectrum of biodiversity of insectivorous and fruit bats. Moreover, the usage of primers for the broad detection of viruses should be reconsidered because geographical variations might possibly affect the sensitivity of primers in a molecular approach.
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@#Prior to COVID-19, dengue was an important public health problem in Malaysia. Due to the movement control order imposed by the Malaysian government to curb the COVID-19 transmission, a study predicted that mosquito-borne diseases would increase during lockdown and partial lockdown seasons. Thus, this study aims to determine the current situation of dengue incidence during the pre-COVID-19 pandemic (2019) and during the COVID-19 pandemic (2020 and 2021). We compared the number of laboratory-confirmed cases in the pre-COVID19 year (2019) and during the COVID-19 pandemic (2020 and 2021). In addition to that, we characterized the clinical manifestation, dengue serotype and viremia levels of dengue patients that were admitted to the Hospital Cancelor Tuanku Muhriz. We found a significant decrease in the number of laboratory-confirmed cases between COVID-19 pandemic and the pre-covid period (p2020=0.064; p2021<0.001). In this study, we found DENV 4 serotype was the most common serotype in dengue patients admitted to our hospital. There was no significant correlation between DENV serotype/viremia level with clinical manifestation of dengue fever and dengue with warning signs. However, patients infected with DENV4 had the highest viral load compared to patients infected with other serotypes. We also found high viremia levels were significantly associated with the febrile phase.
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@#Objective To express dengue virus(DENV)NS2B-NS3 protease in E.coli,optimize the expression conditions and determine the enzyme activity,so as to lay a foundation of screening and discovering of lead compounds targeting DENV.Methods Codon-optimized NS2B-NS3 gene was inserted into pET-28a vector to construct recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3,which was transformed E.coli Rosetta(DE3)competent cells and induced by IPTG to express NS2B-NS3 protease. The optimal expression conditions of NS2B-NS3 protease in E.coli were determined by optimizing induction length,induction temperature and IPTG concentration. NS2B-NS3 protease was isolated and purified by HisTrap~(TM) affinity chromatography column and measured for the protease activity by fluorescence resonance energy transfer(FRET)assay.Results The recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3 was constructed correctly as identified by restriction analysis(NheⅠ/XhoⅠ)and sequencing. The optimal expression conditions of NS2BNS3 protease in E.coli were as follows:induction temperature of 20 ℃,induction length of 10 h and IPTG concentration of0. 2 mmol/L. The purified NS2B-NS3 protease showed a purity of more than 90% with a exhibited a of 20 mg/L,which bound to mouse monoclonal antibody against His-tag specifically and had good hydrolytic activity with a specific activity of 16. 111 U/mg,a K_m of 16. 46 μmol/L and a k_(cat) of 0. 028/s.Conclusion DENV NS2B-NS3 protease with high purity and activity was successfully prepared,which laid an experimental foundation of the establishment of high-throughput screening model for inhibitors targeting NS2B-NS3 protease.
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To screen novel anti-dengue virus (DENV) NS5 RdRp enzyme inhibitors, a series of 5-cyano-2-thiacetoaryl pyrimidinone compounds were designed and synthesized by molecular hybridization method with HCV NS5B RdRp inhibitor 3jc and ZIKV NS5 RdRp inhibitor 4w as lead compounds. The anti-DENV activity of these compounds was evaluated by MTT assay and plaque assay and five compounds showed anti-DENV activity. The most active compound 7a'k showed better anti-DENV activity than that of the positive control ribavirin (EC50 = 7.86 μmol·L-1 vs EC50 = 18.07 μmol·L-1), and the other four compounds showed almost the same anti-DENV activity as ribavirin. Finally, the prediction and simulation of the binding mode through molecular provided new ideas for the further development of this new DENV NS5 RdRp inhibitor.
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ABSTRACT Dengue is a global and growing health threat, especially in Southeast Asia, West Pacific and South America. Infection by the dengue virus (DENV) results in dengue fever, which can evolve to severe forms. Cytokines, especially interferons, are involved in the immunopathogenesis of dengue fever, and so may influence the disease outcomes. The aim of this study was to investigate the association between severe forms of dengue and two single nucleotide polymorphisms (SNPs) in the interferon-gamma gene (IFNG): A256G (rs2069716) and A325G (rs2069727). We included 274 patients infected with DENV serotype 3: 119 cases of dengue without warning signs (DWoWS), and 155 with warning signs (DWWS) or severe dengue (SD). DNA was extracted, and genotyped with Illumina Genotyping Kit or real time PCR (TaqMan probes). We estimated the adjusted Odds Ratios (OR) by multivariate logistic regression models. When comparing with the ancestral AA/AA diplotype (A256G/A325G), we found a protective association of the AA/AG against DWWS/SD among patients with secondary dengue (OR 0.51; 95% IC 0.24-1.10, p = 0.085), adjusting for age and sex. The variant genotype at locus A325G of the IFNG, in combination with the ancestral genotype at locus A256G, can protect against severe clinical forms of secondary dengue in Brazilian DENV3-infected patients.
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Introduction: We previously found that the occurrence of congenital Zika syndrome was under-reported in Mexico. It was postulated that high dengue virus antibody levels found at the population-level in endemic countries might have contributed to the occurrence of the regional pandemic of Zika disease. A case series of suspected cases of congenital Zika syndrome in a maternity hospital in Tuxtla Gutierrez, Chiapas, Mexico was assembled to assess why they were not reported and to explore the hypothesis of dengue virus antibody-dependent enhancement of Zika disease. Methods: We used a quantitative approach to describe clinical and imaging records and used data from interviews of a total of 13 suspected cases of congenital Zika syndrome. We also quantitated dengue 1 and 2 antibodies using an 80% plaque reduction neutralization test of sera specimens obtained from the mothers of these 13 cases and compared them to those of a group of mothers who delivered normal newborns in the same hospital. Results: Only one of the suspected cases was laboratory-confirmed because appropriate specimens were not collected from the newborns as required by the case definition. We found 1) microcephaly, 2) hypoplasia/hypogeneses, thinning or absence of brain structures, 3) multiple birth defects, 4) calcifications, and cysts, 5) meningocele/encephalocele, and 6) hydrocephalus in 100 %, 76.9 %, 38.5 %, 38.5 %, 30.8 %, and 23.1 %, respectively of the case series. The cases clustered geographically, and 77 % occurred between May 2016 to March 2017 and recalled or were told by a doctor they had Zika fever. There was a four-fold increased risk of congenital Zika syndrome among those with dengue 1 antibody as compared to those with dengue 2 antibodies (odds ratio = 3.6; 95 % confidence interval: 0.7, 20.5), reaching only borderline statistical significance. Conclusions: We found in the largest maternal facility of the capital of the State of Chiapas, in Mexico, that only 7.7 % of suspected cases were confirmed, and that the rather complex requirement of cerebrospinal fluid specimens or serological specimens of newborns for suspected cases of congenital Zika syndrome used during the pandemic resulted in low sensitivity of the surveillance system. The finding of higher levels of dengue 1 than dengue 2 antibodies in cases than the referent population, requires further evaluation and may suggest a role for dengue antibody-dependent response in Zika disease.
Introducción: Previamente los autores habían encontrado evidencia de sub-notificación de la ocurrencia del síndrome congénito por Zika en México. Se ha postulado que niveles elevados de anticuerpos contra los virus del dengue a nivel poblacional en los países endémicos hubiese contribuido a la ocurrencia de la pandemia regional de enfermedad por Zika. Ensamblamos una serie de casos sospechosos de síndrome congénito por Zika en un hospital de maternidad en Tuxtla Gutiérrez, Chiapas, México, para evaluar por qué no fueron notificados y explorar la hipótesis de enfermedad por Zika incrementada por anticuerpos anti-dengue. Métodos: Utilizamos un enfoque cuantitativo para describir 13 casos sospechosos de síndrome congénito por revisamos registros clínicos e imágenes, entrevistas. También cuantificamos los niveles de anticuerpos para los virus dengue 1 y 2 en suero de las madres de los casos comparados con los de mujeres que tuvieron recién nacidos normales en el mismo hospital. Resultados: Solamente uno de los 12 casos sospechosos fue confirmado por laboratorio, porque en los demás no se recolectaron especímenes adecuados de los neonatos como lo requería la definición de casos. Encontramos 1) microcefalia, 2) hipoplasia y adelgazamiento de las estructuras cerebrales, 3) malformaciones múltiples, 4) calcificaciones o quistes, 5) meningocele/encefalocele, y 6) hidrocefalia en: 100 %, 76.9 %, 38.5 %, 38.5 %, 30.8 %, y 23.1 %, en ese orden entre los casos sospechosos. Los casos se aglutinaron geográficamente y 77 % ocurrieron entre Mayo del 2016 y Marzo del 2017, y sus madres recordaban que tuvieron o que un profesional de la salud les dijo que tuvieron fiebre por Zika. Encontramos un incremento de casi 4 veces en el riesgo de síndrome congénito por Zika para aquellos con altos niveles de anticuerpos anti-dengue 1 comparado con anticuerpos anti-dengue 2 (cociente de suertes = 3.6; intervalo de confianza del 95 %: 0.7, 20.5), alcanzando solamente una significancia estadística limítrofe. Conclusiones: Encontramos en el establecimiento de atención a la maternidad más grande en la capital de Chiapas, México, que solamente 7.7 % de los casos sospechosos de síndrome congénito por Zika fueron confirmados y que los relativamente complejos requerimientos de la definición de casos de muestras serológicas o de líquido cefalorraquídeo resultó en una baja sensibilidad del sistema de vigilancia. El hallazgo de niveles más altos de anticuerpos a dengue 1 que dengue 2 requiere más evaluación y pudiera sugerir un papel de la respuesta dependiente de anticuerpos al dengue en Zika.
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Background & objectives: Recently, the incidences of chikungunya, dengue and Zika infections have increased due to globalization and urbanization. It is vital that reliable detection tools become available to assess the viral prevalence within mosquito populations. Methods: Based on the previous publications on clinical diagnosis in human infections, for the first time, we described a customized triplex RT-qPCR protocol for simultaneous detection of chikungunya virus (CHIKV), dengue virus serotypes 1-4 (DENV1-4) and Zika virus (ZIKV) in mosquitoes. Results: In preliminary assessment to determine the specificity and sensitivity of primers and probes, all six targets were detected individually with the following thresholds as indicated by calculated pfu equivalents: 3.96x100 for CHIKV, 3.80x101 for DENV1, 3.20x101 for DENV2, 8.00x10-1 for DENV3, 1.58x100 for DENV4, and 6.20x100 for ZIKV. When tested in a full combination of six targets (CDZ mix), CHIKV, DENV1-4 mix or ZIKV were all detected with the thresholds of 1.32x100 for CHIKV, 3.79x100 for DENV1-4 and 2.06x100 for ZIKV. All targets, individually or in full combination were detected in the mixtures of Aedes aegypti (L.) homogenate and viral lysates. A robust evaluation with three replicates in each of three plates for CHIKV, DENV1-4 and ZIKV individually or in full combination was conducted. In individual assays, CHIKV was detected to 3.96x10-1, DENV1-4 to 1.14x100 and ZIKV to 3.20x100 . In full combination assays, CHIKV was detected to 1.32x10-1, DENV1-4 to 3.79x101 and ZIKV to 1.07x100 . Interpretation & conclusion: This triplex RT-qPCR assay appears to consistently detect all six targets and does not cross react with Ae. aegypti homogenate, making it a feasible, practical, and immediately adoptable protocol for use among vector control and other entities, particularly in the endemic areas of CHIKV, DENVs and ZIKV.
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Abstract Acute kidney injury (AKI) is one of the least studied complications of dengue, but it carries high mortality rates and prolonged hospital stay. Due to the severity of this complication, the risk of developing chronic kidney disease (CKD) and the increasing number of dengue cases reported worldwide, particularly in the tropical and subtropical regions of Africa, Southeast Asia and South America, including Brazil, we embarked on this narrative review, aimed to update the epidemiology of AKI associated with dengue, elucidate the main pathophysiological mechanisms of AKI caused by the dengue virus infection, as well as discuss useful information on the prevention and management of AKI in patients with dengue.
Resumo A injúria renal aguda (IRA) é uma das complicações da dengue menos estudadas, mas que acarreta altas taxas de mortalidade e prolongamento do tempo de internação. Devido à gravidade dessa complicação, ao risco de desenvolvimento de doença renal crônica (DRC) e ao número crescente de casos de dengue relatados no mundo, particularmente nas regiões tropicais e subtropicais da África, Sudeste Asiático e América do Sul, incluindo o Brasil, foi proposta esta revisão narrativa que objetivou atualizar a epidemiologia da IRA associada à dengue, elucidar os principais mecanismos fisiopatológicos da IRA causada pela infecção do vírus da dengue, assim como discutir informações úteis sobre a prevenção e o manejo da IRA em pacientes com dengue.