In silico design and validation of a novel multi-epitope vaccine candidate against structural proteins of Chikungunya virus using comprehensive immunoinformatics analyses.

Chikungunya virus (CHIKV) is an emerging viral infectious agent with the potential of causing pandemic. There is neither a protective vaccine nor an approved drug against the virus. The aim of this study was design of a novel multi-epitope vaccine (MEV) candidate against the CHIKV structural proteins using comprehensive immunoinformatics and immune simulation analyses. In this study, using comprehensive immunoinformatics approaches, we developed a novel MEV candidate using the CHIKV structural proteins (E1, E2, 6 K, and E3). The polyprotein sequence was obtained from the UniProt Knowledgebase and saved in FASTA format. The helper and cytotoxic T lymphocytes (HTLs and CTLs respectively) and B cell epitopes were predicted. The toll-like receptor 4 (TLR4) agonist RS09 and PADRE epitope were employed as promising immunostimulatory adjuvant proteins. All vaccine components were fused using proper linkers. The MEV construct was checked in terms of antigenicity, allergenicity, immunogenicity, and physicochemical features. The docking of the MEV construct and the TLR4 and molecular dynamics (MD) simulation were also performed to assess the binding stability. The designed construct was non-allergen and was immunogen which efficiently stimulated immune responses using the proper synthetic adjuvant. The MEV candidate exhibited acceptable physicochemical features. Immune provocation included prediction of HTL, B cell, and CTL epitopes. The docking and MD simulation confirmed the stability of the docked TLR4-MEV complex. The high-level protein expression in the Escherichia coli (E. coli) host was observed through in silico cloning. The in vitro, in vivo, and clinical trial investigations are required to verify the findings of the current study.

Experiências em divulgação científica e sensibilização da população: importância do controle mecânico do vetor Aedes aegypti / Experiences in scientific outreach and public awareness: the importance of mechanical control of the Aedes aegypti vector / Experiencias en divulgación científica y sensibilización de la población: importancia del control mecánico del vector Aedes aegypti

Aedes aegypti é o principal vetor dos agentes etiológicos de dengue, zika e chikungunya, doenças para as quais não existem vacinas totalmente eficazes. Alternativas de controle visando mitigar essas arboviroses são primordiais. Entre essas, o controle mecânico aborda práticas de eliminação e/ou limpeza de criadouros do vetor. Neste relato, apresentamos e avaliamos criticamente ações realizadas pelo grupo, ocorridas entre 2016 e 2019, nas quais divulgamos informação científica clara através do diálogo com a população. Os métodos utilizados foram: 1) palestras em escolas (público infantojuvenil) utilizando slides, fotos e vídeos; 2) oficinas (público misto), estande com material in vivo do ciclo de vida do Aedes, jogos e desenhos. Analisamos dez palestras em escolas do ensino fundamental e médio e vinte oficinas realizadas em diferentes regiões do Brasil. Concluímos que tais ações e suas análises críticas devem ser realizadas continuamente para que sejam bem-sucedidas

Aedes aegypti is the main vector of dengue, zika, and chikungunya etiological agents, diseases for which no effective vaccines are available. Control alternatives aimed at mitigating these arboviruses are essential. Among such, mechanical control addresses practices of elimination and/or cleaning of vector breeding sites. Here, we presented and critically evaluated actions carried out by ourselves. These actions took place between 2016 and 2019, where we disseminated clear scientific information through dialogue with the population. The following methods were employed: 1) lectures in schools (children and youth audiences) using slides, photos, and videos; 2) workshops (mixed audience), stand with in vivo material from the Aedeslife cycle, games, and drawings. Ten lectures in elementary and high schools and twenty workshops held in different regions of Brazil were analyzed. It was concluded that such actions and their critical analyzes must be carried out continuously to be successful

Aedes aegypti es el principal vector de los agentes etiológicos del dengue, zika y chikungunya, enfermedades para las que no existen vacunas totalmente eficaces. Las alternativas de control para mitigar estas arbovirosis son fundamentales. El control mecánico, representa una de estas alternativas, aborda prácticas de eliminación y/o limpieza de criaderos del vector. En este informe presentamos y evaluamos de manera crítica las acciones realizadas por el grupo entre los años 2016 y 2019. Presentamos información científica clara a través del diálogo con la población mediante los siguientes métodos: 1) conferencias en escuelas (público infantil) utilizando diapositivas, fotos y videos; 2) Talleres (público mixto), stand con material in vivo del ciclo de vida del Aedes, juegos y dibujos. Analizamos diez conferencias en escuelas (primarias y secundarias) y veinte talleres realizados en diferentes regiones de Brasil. Concluimos que tales acciones y el análisis crítico de las mismas deben llevarse a cabo de manera continua para que resulten exitosas.

Perfil das notificações de dengue e febre chikungunya em Cabo Frio/Rj, 2014-2019 / Profile of dengue and chikungunya notifications in Cabo Frio/Rj, 2014-2019 / Perfil de las notificaciones de dengue y chikungunya en Cabo Frio/Rj, 2014-2019

Introdução: De acordo com a literatura científica, diagnósticos clínicos diferenciais de arboviroses representam uma dificuldade no que tange à dengue, na medida em que está no Brasil há muitos anos, o que acarreta em ser a arbovirose mais conhecida no país. As notificações de arboviroses se tornaram obrigatórias para inserção no SINAN, possibilitando a construção de perfis demográficos e o cálculo de incidências a partir de informações específicas para estas doenças. No que tange à dengue, a epidemia deste agravo ocorre no país desde 1986, evidenciando falhas na prevenção, relacionadas a aspectos socioeconômicos e ambientais. Objetivo: analisar perfis das notificações de dengue e febre de chikungunya dos casos notificados no município de Cabo Frio. Metodologia: Trata-se de estudo transversal e descritivo, com uso de dados secundários do SINAN referentes a casos de arboviroses no município de Cabo Frio/RJ. Foram observadas variáveis relacionadas ao sexo, escolaridade, raça/cor e critérios de confirmação, além do grau de completude. Resultados: Foram notificados 8.777 casos suspeitos de arboviroses, incluindo-se 1.367 notificações (15,57%) referentes à febre de chikungunya e 1.986 (22,63%), à dengue. Em relação ao desfecho, 1186 casos (51,45%) foram fechados como inconclusivos e 344 destes (14,92%) foram descartados como arboviroses. Dentre os inconclusivos, 943 (79,51%) eram referentes à notificação de dengue, idem para os 277 casos descartados (80,52%). Conclusão: Observou-se baixa taxa de completude nas fichas de notificação, explicada pelo baixo número de recursos humanos e pela insuficiente infraestrutura. Sugere-se a interação de diferentes profissionais e pesquisadores, facilitando a compreensão da complexa dinâmica das arboviroses em questão.

Introduction: According to the scientific literature, differential clinical diagnoses of arboviruses represent a difficulty with regard to dengue, as it has been present in Brazil for many years, which makes it the most well-known arbovirus in the country. Notifications of arboviruses became mandatory for inclusion in SINAN, enabling the construction of demographic profiles and the calculation of incidences based on specific information for these diseases. With regard to dengue, the epidemic of this disease has occurred in the country since 1986, showing failures in prevention, related to socioeconomic and environmental aspects. Objective: to analyze profiles of notifications of dengue and chikungunya fever of cases notified in the municipality of Cabo Frio. Methodology: This is a cross-sectional and descriptive study, using secondary data from SINAN regarding cases of arboviruses in the municipality of Cabo Frio/RJ. Variables related to sex, education, race/color and confirmation criteria were observed, in addition to the degree of completeness. Results: 8,777 suspected cases of arboviruses were reported, including 1,367 reports (15.57%) referring to chikungunya fever and 1,986 (22.63%) to dengue fever. Regarding the outcome, 1186 cases (51.45%) were closed as inconclusive and 344 of these (14.92%) were discarded as arboviruses. Among the inconclusive ones, 943 (79.51%) were related to dengue notification, the same for the 277 discarded cases (80.52%). Conclusion: A low completeness rate was observed in the notification forms, explained by the low number of human resources and insufficient infrastructure. It is suggested the interaction of different professionals and researchers, facilitating the understanding of the complex dynamics of the arboviruses in question.

Introducción: Según la literatura científica, los diagnósticos clínicos diferenciales de los arbovirus representan una dificultad con respecto al dengue, ya que está presente en Brasil desde hace muchos años, lo que lo convierte en el arbovirus más conocido en el país. Las notificaciones de arbovirus pasaron a ser obligatorias para su inclusión en el SINAN, lo que permitió la construcción de perfiles demográficos y el cálculo de incidencias a partir de información específica de estas enfermedades. Con respecto al dengue, la epidemia de esta enfermedad se presenta en el país desde 1986, mostrando fallas en la prevención, relacionadas con aspectos socioeconómicos y ambientales. Objetivo: analizar perfiles de notificaciones de dengue y fiebre chikungunya de los casos notificados en el municipio de Cabo Frio. Metodología: Se trata de un estudio transversal y descriptivo, utilizando datos secundarios del SINAN sobre casos de arbovirus en el municipio de Cabo Frio/RJ. Se observaron variables relacionadas con el sexo, escolaridad, raza/color y criterios de confirmación, además del grado de completitud. Resultados: se notificaron 8.777 casos sospechosos de arbovirus, de los cuales 1.367 (15,57%) se referían a fiebre chikungunya y 1.986 (22,63%) a dengue. En cuanto al resultado, 1186 casos (51,45%) se cerraron como no concluyentes y 344 de estos (14,92%) se descartaron como arbovirus. Entre los inconclusos, 943 (79,51%) estaban relacionados con la notificación de dengue, lo mismo para los 277 casos descartados (80,52%). Conclusión: Se observó un bajo índice de completitud en los formularios de notificación, explicado por el bajo número de recursos humanos y la infraestructura insuficiente. Se sugiere la interacción de diferentes profesionales e investigadores, facilitando la comprensión de la compleja dinámica de los arbovirus en cuestión.

Construction and Evaluation of Recombinant Adenovirus Candidate Vaccines for Chikungunya Virus.

Chikungunya virus (CHIKV) is a mosquito-borne virus. The emergence of CHIKV infection has raised global concern, and there is a growing need to develop safe and effective vaccines. Here, adenovirus 5 was used as the vaccine vector to construct recombinant adenoviruses expressing CHIKV E2, E1, and E2-6K-E1, respectively. And then the immunogenicity and protective efficiency against CHIKV were evaluated in BALB/c mice. Compared to the ad-wt control group, all three vaccines elicited significant humoral and cellar immune responses. The levels of neutralizing antibodies in the rAd-CHIKV-E2-6K-E1 and rAd-CHIKV-E2 groups both reached 1:256, which were 3.2 times higher than those in the rAd-CHIKV-E1 group. Furthermore, the levels of lymphocyte proliferation in rAd-CHIKV-E2-6K-E1 group were the highest. Besides, the concentrations of IFN-γ and IL-4 in mice immunized with rAd-CHIKV-E2-6K-E1 were 1.37 and 1.20 times higher than those in ad-wt immunized mice, respectively. After the challenge, mice in the rAd-CHIKV-E2-6K-E1 and rAd-CHIKV-E2 groups lost 2% of their body weight compared with 5% in the ad-wt control group. And low viral loads were detected in the heart, kidney, and blood of mice immunized with rAd-CHIKV-E2-6K-E1 and rAd-CHIKV-E2 at 3-5 dpc, which decreased by 0.4-0.7 orders of magnitude compared with the ad-wt control. Overall, these data suggest that the recombinant adenovirus is a potential candidate vaccine against CHIKV.

An mRNA vaccine encoding Chikungunya virus E2-E1 protein elicits robust neutralizing antibody responses and CTL immune responses.

Arthropod-borne chikungunya virus (CHIKV) infection can cause a debilitating arthritic disease in human. However, there are no specific antiviral drugs and effective licensed vaccines against CHIKV available for clinical use. Here, we developed an mRNA-lipid nanoparticle (mRNA-LNP) vaccine expressing CHIKV E2-E1 antigen, and compared its immunogenicity with soluble recombinant protein sE2-E1 antigen expressed in S2 cells. For comparison, we first showed that recombinant protein antigens mixed with aluminum adjuvant elicit strong antigen-specific humoral immune response and a moderate cellular immune response in C57BL/6 mice. Moreover, sE2-E1 vaccine stimulated 12-23 folds more neutralizing antibodies than sE1 vaccine and sE2 vaccine. Significantly, when E2-E1 gene was delivered by an mRNA-LNP vaccine, not only the better magnitude of neutralizing antibody responses was induced, but also greater cellular immune responses were generated, especially for CD8+ T cell responses. Moreover, E2-E1-LNP induced CD8+ T cells can perform cytotoxic effect in vivo. Considering its better immunogenicity and convenience of preparation, we suggest that more attention should be placed to develop CHIKV E2-E1-LNP mRNA vaccine.

Household clustering supports a novel chemoprophylaxis trial design for a mosquito-borne viral disease.

Infections because of chikungunya and other mosquito-borne viruses, such as dengue and Zika, represent an area of significant unmet medical need. There are currently no approved medicines for prophylaxis or treatment of these diseases, and the development and implementation of vaccines against these viruses have proved problematic. Although antiviral molecules with treatment and prophylactic potential against the chikungunya virus have been identified, no successful field trials have been reported. Chemoprophylaxis may be attractive for unvaccinated at-risk populations; however, performing a successful chemoprophylaxis trial during a chikungunya outbreak will require a clearly identifiable at-risk population. We propose the application of a household transmission model as used in testing drugs against respiratory viruses. Current evidence on household clustering of chikungunya and other Aedes mosquito-borne viral infections is supportive. We suggest that this model may improve prophylaxis trial feasibility and focus research and future treatment on a population likely to benefit.

Percepção da comunidade sobre suas ações preventivas contra dengue, zika e chikungunya nas cinco regiões do Brasil / Community perception of its preventive actions against dengue, zika and chikungunya in five Brazilian regions

Resumo O controle ambiental do vetor no interior e ao redor dos domicílios, enquanto estratégia fundamental para a prevenção de arboviroses como dengue, zika e chikungunya, demanda um envolvimento ininterrupto da população. A pandemia de Covid-19 e, consequentemente, o isolamento social necessário ao seu controle, têm reduzido a vigilância profissional nos domicílios, tornando a atuação da população ainda mais necessária. Estudos que buscam compreender o envolvimento das comunidades na execução das ações preventivas têm sido quase exclusivamente locais. O artigo relata uma pesquisa-intervenção baseada em oficinas sobre práticas preventivas realizadas antes do isolamento social, em 16 municípios das 5 regiões brasileiras, as quais envolveram um total de 379 participantes. Os resultados apresentam as ações preventivas preconizadas por campanhas que são as mais comumente compreendidas e executadas pela população; as situações que dificultam a execução; as práticas preventivas de iniciativa própria dos cidadãos; e as práticas baseadas em saberes populares. Ficou evidente que as informações fornecidas às comunidades não podem estar restritas às campanhas de massa, e que se faz necessário investir em ações educativas mais efetivas e adequadas à variedade de contextos nacionais, com vistas a construções coletivas e intersetoriais de estratégias de enfrentamento às arboviroses.

Abstract The environmental control of the vector inside and around households, as a fundamental strategy for the prevention of arboviruses such as dengue, zika and chikungunya, demands an uninterrupted involvement of the population. The pandemic context of Covid-19, and consequently the social isolation has reduced professional surveillance in the households, making the performance of the population even more necessary. Studies that seek to understand the involvement of communities in the implementation of preventive actions have been almost exclusively local. This article reports an intervention research based on workshops on preventive practices carried out before social isolation, in 16 municipalities in the five Brazilian regions and involved 379 participants. Our results show the preventive actions recommended by campaigns that are the most commonly understood and performed by the population; situations that hinder execution; preventive practices of citizens' own initiative; and practices based on popular knowledge. It became evident that information to communities cannot be restricted to mass campaigns, and that it is necessary to invest in educational actions adequate to the variety of national contexts, seeking collective and intersectoral constructions of strategies to confront arboviruses.

In Silico Identification of Chikungunya Virus B- and T-Cell Epitopes with High Antigenic Potential for Vaccine Development.

Reverse vaccinology is an outstanding strategy to identify antigens with high potential for vaccine development. Different parameters of five prediction programs were used to assess their sensitivity and specificity to identify B-cell epitopes of Chikungunya virus (CHIKV) strains reported in the IEDB database. The results, based on the use of 15 to 20 mer epitopes and the polyproteins to which they belong, were compared to establish the best parameters to optimize the prediction of antigenic peptides of the Mexican strain CHIKV AJV21562.1. LBtope showed the highest specificity when we used the reported epitopes and polyproteins but the worst sensitivity with polyproteins; ABCpred had similar specificity to LBtope only with the epitopes reported and showed moderate specificity when we used polyproteins for the predictions. Because LBtope was more reliable in predicting true epitopes, it was used as a reference program to predict and select six novel epitopes of the Mexican strain of CHIKV according to prediction frequency, viral genome localization, and non-homology with the human proteome. On the other hand, six bioinformatics programs were used with default parameters to predict T-cell epitopes in the CHIKV strains AJV21562.1 and AJV21561.1. The sequences of the polyproteins were analyzed to predict epitopes present in the more frequent HLA alleles of the Mexican population: DQA1*03011, DQA1*0401, DQA1*0501, DQB1*0201, DQB1*0301, DQB1*0302, and DQB1*0402. Fifteen predicted epitopes in the non-structural and 15 predicted epitopes in the structural polyprotein (9- to 16-mers) with the highest scores of each allele were compared to select epitopes with at least 80% identity. Next, the epitopes predicted with at least two programs were aligned to the human proteome, and 12 sequences without identity with the human proteome were identified as potential antigenic candidates. This strategy would be useful to evaluate vaccine candidates against other viral diseases affecting the countries of the Americas and to increase knowledge about these diseases.

Innovación Educativa: Aprendizaje lúdico para el control de arbovirosis / Educational Innovation: Playful learning for arbovirus control

Las arboviroris, son enfermedades transmitidas por artrópodos (insectos vectores), presentan una clínica habitual de fiebre, síntomas articulares, hemorrágicos y neurológicos. Los de mayor importancia en salud pública son el zika, dengue y chikungunya, y se transmiten a las personas principalmente por la picadura de un mosquito de la especie Aedes. Para resolver la falta de información en la población joven para el control y prevención de estas enfermedades vectoriales se plantea utilizar nuevas estrategias pedagógicas y tecnologías lúdicas, como los videojuegos, para una innovación educativa que promueva el aprendizaje. Se realizó una investigación descriptiva de cohorte transversal, en escolares de 5 y 6 grado, en tres colegios primarios, de la capital de la República del Perú. La muestra estuvo integrada por un total de 288 escolares y fue dividida en cuatro (4) grupos de 6 niños cada uno para la aplicación de la innovación educativa, asignados como: (ETE), (CIE 1), (CIE 2) y (CIE 3). La evaluación del conocimiento, hábitos y aceptación de los métodos se ejecutó pre y post-intervención, midiéndose mediante encuestas. Por consiguiente luego de los resultados en promedio post-intervención se concluyó que la captación de conocimientos y generación de hábitos fueron mayores para los grupos que utilizaron los videojuegos como innovación educativa lúdica, siendo el videojuego interactivo "Pica el Aedes", el que obtuvo el mayor porcentaje de aprendizaje. Sin embargo el videojuego interactivo "Pueblo Pitanga, enemigos silenciosos" fue el que obtuvo la mayor aceptación por parte de los escolares(AU)

The arboviroris, are diseases transmitted by arthropods (vector insects), they present a habitual clinic of fever, joint, hemorrhagic and neurological symptoms. The most important ones in public health are Zika, dengue and chikungunya, and they are transmitted to people mainly through the bite of an Aedes species mosquito. To resolve the lack of information in the young population for the control and prevention of these vector diseases, it is proposed to use new pedagogical strategies and recreational technologies, such as video games, for an educational innovation that promotes learning. A descriptive cross-sectional cohort investigation was conducted in 5th and 6th grade schoolchildren in three primary schools in the capital of the Republic of Peru. The sample consisted of a total of 288 schoolchildren and was divided into four (4) groups of 6 children each for the application of educational innovation, assigned as: (ETE), (CIE 1), (CIE 2) and (CIE 3). The evaluation of the knowledge, habits and acceptance of the methods was carried out before and after the intervention, being measured by means of surveys. Consequently, after the average post-intervention results, it was concluded that the uptake of knowledge and generation of habits were higher for the groups that used video games as a recreational educational innovation, being the interactive video game "Pica el Aedes", the one that obtained the higher percentage of learning. However, the interactive video game "Pueblo Pitanga, enemigos silenciosos" was the one that obtained the greatest acceptance from schoolchildren(AU)

MK2a inhibitor CMPD1 abrogates chikungunya virus infection by modulating actin remodeling pathway.

Chikungunya virus (CHIKV) epidemics around the world have created public health concern with the unavailability of effective drugs and vaccines. This emphasizes the need for molecular understanding of host-virus interactions for developing effective targeted antivirals. Microarray analysis was carried out using CHIKV strain (Prototype and Indian) infected Vero cells and two host isozymes, MAPK activated protein kinase 2 (MK2) and MAPK activated protein kinase 3 (MK3) were selected for further analysis. The substrate spectrum of both enzymes is indistinguishable and covers proteins involved in cytokines production, endocytosis, reorganization of the cytoskeleton, cell migration, cell cycle control, chromatin remodeling and transcriptional regulation. Gene silencing and drug treatment were performed in vitro and in vivo to unravel the role of MK2/MK3 in CHIKV infection. Gene silencing of MK2 and MK3 abrogated around 58% CHIKV progeny release from the host cell and a MK2 activation inhibitor (CMPD1) treatment demonstrated 68% inhibition of viral infection suggesting a major role of MAPKAPKs during late CHIKV infection in vitro. Further, it was observed that the inhibition in viral infection is primarily due to the abrogation of lamellipodium formation through modulation of factors involved in the actin cytoskeleton remodeling pathway. Moreover, CHIKV-infected C57BL/6 mice demonstrated reduction in the viral copy number, lessened disease score and better survivability after CMPD1 treatment. In addition, reduction in expression of key pro-inflammatory mediators such as CXCL13, RAGE, FGF, MMP9 and increase in HGF (a CHIKV infection recovery marker) was observed indicating the effectiveness of the drug against CHIKV. Taken together it can be proposed that MK2 and MK3 are crucial host factors for CHIKV infection and can be considered as important target for developing effective anti-CHIKV strategies.

Gestión integrada para la prevención y control del dengue y otras arbovirosis en la Municipalidad de Ambato / Integrated management for the prevention and control of dengue and other arboviruses in the Municipality of Ambato

Las arbovirosis por Dengue (VD), Zika (VZIK) y Chikungunya (VCHIK), transmitidas por vectores del género Aedes, tienen alta prevalencia en países tropicales, especialmente en Ecuador donde se les considera problemas de salud pública. Se realizó un estudio descriptivo para identificar el nivel de cumplimiento de la estrategia de gestión integrada (EGI) para prevención y control de VD y otros arbovirus en Ambato, Tungurahua - Ecuador. Se realizó perfil comunitario a partir de fuentes oficiales y secundarias, se evaluó componentes del ambiente mediante lista de verificación y se evaluó cumplimiento de EGI mediante encuesta al personal del sector salud. Datos consultados señalan que en la provincia existe 36% y 10% de pobreza y pobreza extrema, respectivamente; además adecuado servicio de agua potable, recepción de agua por tubería, gestión de residuos sólidos y servicio de electricidad en ambas zonas, urbana y rural. Se constató que más del 50% de la población rural presenta condiciones óptimas de salud ambiental: aire limpio (90,28%) y agua potable (66,94%); en la población urbana destaca higiene de alimentos (63,89%) y microambiente doméstico (69,44%), con deficiencia en recolección de residuos sólidos (33,89%). La EGI tuvo un nivel de cumplimiento de 100% en parroquias urbanas en componentes laboratorio, atención al paciente, promoción y comunicación, mientras que en parroquias rurales, salud ambiental tuvo 63% e investigación y capacitación menor nivel (47,78%); siendo necesario mejorar sistemas de desechos sólidos en la población urbana y promover el conocimiento y capacitación epidemiológica en la rural, para reducir el riesgo de transmisión por arbovirus(AU)

Dengue (DV), Zika (ZIKV) and Chikungunya (CHIKV) arboviruses, transmitted by vectors of the Aedes genus, have a high prevalence in tropical countries, especially in Ecuador where they are considered public health problems. A descriptive study was carried out to identify the level of compliance with the integrated management strategy (EGI) for prevention and control of DV and other arboviruses in Ambato, Tungurahua - Ecuador. A community profile was made from official and secondary sources, components of the environment were evaluated by means of a checklist and compliance with EGI was evaluated by means of a survey of health sector personnel. Consulted data indicate that in the province there is 36% and 10% of poverty and extreme poverty, respectively; In addition, adequate drinking water service, receiving water through pipes, solid waste management and electricity service in both urban and rural areas. It was found that more than 50% of the rural population presents optimal environmental health conditions: clean air (90.28%) and drinking water (66.94%); In the urban population, food hygiene (63.89%) and domestic microenvironment (69.44%) stand out, with a deficiency in solid waste collection (33.89%). The EGI had a compliance level of 100% in urban parishes in laboratory, patient care, promotion and communication components, while in rural parishes, environmental health had 63% and research and training lower level (47.78%); being necessary to improve solid waste systems in the urban population and promote epidemiological knowledge and training in rural areas, to reduce the risk of transmission by arbovirus(AU)

A single dose of ChAdOx1 Chik vaccine induces neutralizing antibodies against four chikungunya virus lineages in a phase 1 clinical trial.

Chikungunya virus (CHIKV) is a reemerging mosquito-borne virus that causes swift outbreaks. Major concerns are the persistent and disabling polyarthralgia in infected individuals. Here we present the results from a first-in-human trial of the candidate simian adenovirus vectored vaccine ChAdOx1 Chik, expressing the CHIKV full-length structural polyprotein (Capsid, E3, E2, 6k and E1). 24 adult healthy volunteers aged 18-50 years, were recruited in a dose escalation, open-label, nonrandomized and uncontrolled phase 1 trial (registry NCT03590392). Participants received a single intramuscular injection of ChAdOx1 Chik at one of the three preestablished dosages and were followed-up for 6 months. The primary objective was to assess safety and tolerability of ChAdOx1 Chik. The secondary objective was to assess the humoral and cellular immunogenicity. ChAdOx1 Chik was safe at all doses tested with no serious adverse reactions reported. The vast majority of solicited adverse events were mild or moderate, and self-limiting in nature. A single dose induced IgG and T-cell responses against the CHIKV structural antigens. Broadly neutralizing antibodies against the four CHIKV lineages were found in all participants and as early as 2 weeks after vaccination. In summary, ChAdOx1 Chik showed excellent safety, tolerability and 100% PRNT50 seroconversion after a single dose.

Human IFITM3 restricts chikungunya virus and Mayaro virus infection and is susceptible to virus-mediated counteraction.

Interferon-induced transmembrane (IFITM) proteins restrict membrane fusion and virion internalization of several enveloped viruses. The role of IFITM proteins during alphaviral infection of human cells and viral counteraction strategies are insufficiently understood. Here, we characterized the impact of human IFITMs on the entry and spread of chikungunya virus and Mayaro virus and provide first evidence for a CHIKV-mediated antagonism of IFITMs. IFITM1, 2, and 3 restricted infection at the level of alphavirus glycoprotein-mediated entry, both in the context of direct infection and cell-to-cell transmission. Relocalization of normally endosomal IFITM3 to the plasma membrane resulted in loss of antiviral activity. rs12252-C, a naturally occurring variant of IFITM3 that may associate with severe influenza in humans, restricted CHIKV, MAYV, and influenza A virus infection as efficiently as wild-type IFITM3 Antivirally active IFITM variants displayed reduced cell surface levels in CHIKV-infected cells involving a posttranscriptional process mediated by one or several nonstructural protein(s) of CHIKV. Finally, IFITM3-imposed reduction of specific infectivity of nascent particles provides a rationale for the necessity of a virus-encoded counteraction strategy against this restriction factor.

Evaluation of neutralizing antibodies produced by papaya mosaic virus nanoparticles fused to the E2EP3 peptide epitope of Chikungunya envelope.

Chikungunya virus (CHIKV) infection is the cause of acute symptoms and chronic symmetrical polyarthritis associated with long-term morbidity and mortality. Currently, there is no available licensed vaccine or particularly useful drug for human use against CHIKV infection. This study was conducted to evaluate the efficacy of antibodies produced by papaya mosaic virus (PapMV) nanoparticles fused to E2EP3 peptide of CHIKV envelope as a recombinant CHIKV vaccine. PapMV, PapMV-C- E2EP3, and E2EP3-N-PapMV were produced in E. coli with an approximate size of 27 to 30 kDa. ICR mice (5 to 6 weeks of age) were injected subcutaneously with 25 micrograms of vaccine construct, and ELISA measured the titer of CHIKV specific IgG antibodies. The results showed that both recombinant proteins E2EP3-N-PapMV and PapMVC-E2EP3 were able to induce IgG antibodies production in immunized mice against CHIKV while immunization with recombinant PapMV showed no IgG antibodies induction. The neutralizing activity of the antibodies generated by either E2EP3-N-PapMV or PapMV-C-E2EP3 exhibited similar inhibition to CHIKV replication in Vero cells using the cells based antibody neutralizing assay and analyzed by plaque formation assay. This study showed the effectiveness of nanoparticles vaccine generated by fusing epitope peptide of CHIKV envelope to papaya mosaic virus envelope in inducing a robust immune response in mice against CHIKV. The data showed that levels of neutralizing antibodies correlate with a protective immune response CHIKV replication.

Non-replicating adenovirus based Mayaro virus vaccine elicits protective immune responses and cross protects against other alphaviruses.

Mayaro virus (MAYV) is an alphavirus endemic to South and Central America associated with sporadic outbreaks in humans. MAYV infection causes severe joint and muscle pain that can persist for weeks to months. Currently, there are no approved vaccines or therapeutics to prevent MAYV infection or treat the debilitating musculoskeletal inflammatory disease. In the current study, a prophylactic MAYV vaccine expressing the complete viral structural polyprotein was developed based on a non-replicating human adenovirus V (AdV) platform. Vaccination with AdV-MAYV elicited potent neutralizing antibodies that protected WT mice against MAYV challenge by preventing viremia, reducing viral dissemination to tissues and mitigating viral disease. The vaccine also prevented viral-mediated demise in IFN⍺R1-/- mice. Passive transfer of immune serum from vaccinated animals similarly prevented infection and disease in WT mice as well as virus-induced demise of IFN⍺R1-/- mice, indicating that antiviral antibodies are protective. Immunization with AdV-MAYV also generated cross-neutralizing antibodies against two related arthritogenic alphaviruses-chikungunya and Una viruses. These cross-neutralizing antibodies were protective against lethal infection in IFN⍺R1-/- mice following challenge with these heterotypic alphaviruses. These results indicate AdV-MAYV elicits protective immune responses with substantial cross-reactivity and protective efficacy against other arthritogenic alphaviruses. Our findings also highlight the potential for development of a multi-virus targeting vaccine against alphaviruses with endemic and epidemic potential in the Americas.

Adenoviral-Vectored Mayaro and Chikungunya Virus Vaccine Candidates Afford Partial Cross-Protection From Lethal Challenge in A129 Mouse Model.

Mayaro (MAYV) and chikungunya viruses (CHIKV) are vector-borne arthritogenic alphaviruses that cause acute febrile illnesses. CHIKV is widespread and has recently caused large urban outbreaks, whereas the distribution of MAYV is restricted to tropical areas in South America with small and sporadic outbreaks. Because MAYV and CHIKV are closely related and have high amino acid similarity, we investigated whether vaccination against one could provide cross-protection against the other. We vaccinated A129 mice (IFNAR -/-) with vaccines based on chimpanzee adenoviral vectors encoding the structural proteins of either MAYV or CHIKV. ChAdOx1 May is a novel vaccine against MAYV, whereas ChAdOx1 Chik is a vaccine against CHIKV already undergoing early phase I clinical trials. We demonstrate that ChAdOx1 May was able to afford full protection against MAYV challenge in mice, with most samples yielding neutralizing PRNT80 antibody titers of 1:258. ChAdOx1 May also provided partial cross-protection against CHIKV, with protection being assessed using the following parameters: survival, weight loss, foot swelling and viremia. Reciprocally, ChAdOx1 Chik vaccination reduced MAYV viral load, as well as morbidity and lethality caused by this virus, but did not protect against foot swelling. The cross-protection observed is likely to be, at least in part, secondary to cross-neutralizing antibodies induced by both vaccines. In summary, our findings suggest that ChAdOx1 Chik and ChAdOx1 May vaccines are not only efficacious against CHIKV and MAYV, respectively, but also afford partial heterologous cross-protection.

Avaliação da armadilha ovitrampa iscada com atraente natural para o monitoramento de Aedes spp. em Dili, capital do Timor-Leste / Evaluation of the baited ovitrap with natural attractant for monitoring Aedes spp. in Dili, capital of East Timor

Resumo A dengue ocorre no Timor-Leste desde 2005, porém não existe um programa de monitoramento e controle do "Aedes aegypti". O objetivo deste estudo foi de avaliar a armadilha ovitrampa iscada com atraente natural como uma possível ferramenta para monitorar o vetor das arboviroses: Dengue (DENV), Chikungunya (CHIKV) e Zika (ZIKV). O estudo foi realizado na cidade de Dili, capital do Timor-Leste, entre as semanas epidemiológicas 32 (02/08) a 48 (02/12) de 2016. Foram instaladas 70 armadilhas Ovitrampa, em residências de 15 sucos (ruas), de quatro Postos Administrativos (bairros) da cidade. Para as analises dos dados utilizou-se os indicadores entomológicos: Índice de Positividade de Ovitrampa (IPO), Índice de Densidade Vetorial (IDV) e Índice de Densidade de Ovos (IDO). Durante o experimento foram coletados 158.904 ovos de Aedes spp.. O IPO demonstrou que todas as áreas tiveram 98% a 100% de armadilhas contendo ovos de Aedes spp.. Os indicadores IDO e IPO apresentaram correlações positivas e significativa com a temperatura. A defasagem de duas e três semanas para precipitação indicou correlação positiva significativa para IDV e IDO. Portanto, a armadilha ovitrampa é uma ferramenta que pode integrar as ações de um programa de monitoramento e controle de Aedes spp. no Timor-Leste.

Avaliação da qualidade metodológica de diretrizes de vigilância e manejo clínico de dengue e chikungunya / Methodological quality assessment of guidelines for surveillance and clinical management of dengue and chikungunya / Evaluación de la calidad metodológica de directrices de vigilancia y manejo clínico del dengue y chikungunya

Resumo: A pesquisa objetivou avaliar a qualidade metodológica de diretrizes do Ministério da Saúde brasileiro, da Organização Pan-Americana da Saúde (OPAS) e da Organização Mundial da Saúde (OMS) sobre vigilância e manejo clínico de dengue e chikungunya. Trata-se de um estudo descritivo, no qual a ferramenta Appraisal of Guidelines for Research & Evaluation Reporting Checklist II (AGREE II) foi aplicada por quatro avaliadores, de forma independente e mascarada, para seis diretrizes. Cada avaliador atribuiu um escore de 1 (discordo totalmente) a 7 (concordo completamente) aos 23 itens dos domínios do AGREE II: escopo e propósito; envolvimento das partes interessadas; rigor no desenvolvimento; clareza da apresentação; aplicabilidade e independência editorial. As diretrizes de dengue da OPAS (média = 5,2, DP = 0,8) e da OMS (média = 4,5, DP = 0,5) obtiveram maiores pontuações globais, sendo recomendadas com modificações por todos os avaliadores, e a do Ministério da Saúde (média = 2,7, DP = 0,4) não foi recomendada por um deles. Já as pontuações das diretrizes de chikungunya foram baixas (médias variando de 2,2 a 3,0) independentemente do órgão que as elaborou. Os domínios com maior conformidade foram "clareza da apresentação" (mediana de 84,7%) e "escopo e propósito" (77,1%), e os de menor conformidade foram "independência editorial" (5,2%) e "rigor no desenvolvimento" (9,1%). O estudo identificou lacunas na qualidade metodológica das diretrizes relacionadas, principalmente, à transparência nos processos de busca, seleção das evidências científicas e formulação das recomendações, além de falta de clareza quanto ao financiamento e possíveis conflitos de interesses.

Abstract: The study aimed to assess the methodological quality of guidelines by the Brazilian Ministry of Health, Pan American Health Organization (PAHO), and World Health Organization (WHO) on surveillance and clinical management of dengue and chikungunya. This was a descriptive study in which the tool Appraisal of Guidelines for Research & Evaluation Reporting Checklist II (AGREE II) was applied by four evaluators in independent and masked fashion for six guidelines. Each evaluator assigned a score from 1 (disagree completely) to 7 (agree completely) to the 23 items in the AGREE II domains: scope and purpose; stakeholder involvement; rigor in the development; clarity of presentation; applicability; and editorial independence. The dengue guidelines by PAHO (mean = 5.2, SD = 0.8) and WHO (mean = 4.5, SD = 0.5) obtained the highest overall scores and were recommended with modifications by all the evaluators, while the Brazilian Ministry of Health guidelines (mean = 2.7, SD = 0.4) were not recommended by any of them. Meanwhile, the chikungunya guidelines scored low (means from 2.2 to 3.0) for all three agencies. The domains with the greatest conformity were "clarity of presentation" (median 84.7%) and "scope and purpose" (77.1%), while those with the lowest conformity were "editorial independence" (5.2%) and "rigor in development" (9.1%). The study identified gaps in the guidelines' methodological quality, mainly in transparency of the work processes, selection of scientific evidence, and formulation of recommendations, besides lack of clarity in financing and possible conflicts of interest.

Resumen: El objetivo de la investigación fue evaluar la calidad metodológica de las directrices del Ministerio de Salud (MS) brasileño, de la Organización Panamericana de la Salud (OPAS) y de la Organización Mundial de la Salud (OMS) sobre vigilancia y manejo clínico del dengue y chikungunya. Se trata de un estudio descriptivo, en el cual la herramienta Appraisal of Guidelines for Research & Evaluation Reporting Checklist II (AGREE II) fue aplicada por parte de cuatro evaluadores, de forma independiente y oculta, en seis directrices. Cada evaluador atribuyó una puntuación de 1 (en desacuerdo totalmente) a 7 (concuerdo completamente) a los 23 ítems de los dominios del AGREE II: alcance y propósito; implicando las partes interesadas; rigor en el desarrollo; claridad de la presentación; aplicabilidad e independencia editorial. Las directrices del dengue de la OPAS (media = 5,2, DP = 0,8) y OMS (media = 4,5, DP = 0,5) obtuvieron mayores puntuaciones globales, siendo recomendadas con modificaciones por todos los evaluadores, respecto a las del Ministerio de Salud (media = 2,7, DP = 0,4) no se recomendó por parte de uno de ellos. Ya las puntuaciones de las directrices de chikungunya fueron bajas (medias variando de 2,2 a 3,0), independientemente del órgano que las elaboró. Los dominios con mayor conformidad fueron "clareza de la presentación" (media de 84,7%) y "alcance y propósito" (77,1%), mientras que los de menor conformidad fueron "independencia editorial" (5,2%) y "rigor en el desarrollo" (9,1%). El estudio identificó lagunas en la calidad metodológica de las directrices relacionadas, principalmente, respecto a la transparencia en los procesos de búsqueda, selección de las evidencias científicas y formulación de las recomendaciones, además de la falta de claridad respecto a la financiación y posibles conflictos de intereses.

Gestão adequada de resíduos sólidos como fator de proteção na ocorrência da dengue / Adequate solid waste management as a protection factor against dengue cases / Gestión adecuada de los residuos sólidos como factor de protección contra los casos de dengue

RESUMO Objetivo. Verificar a existência de associação de indicadores de gestão de resíduos sólidos e socioeconômicos municipais com índices de incidência de dengue, Zika e Chikungunya nos municípios do estado brasileiro de Minas Gerais. Métodos. Este estudo de caráter exploratório, quantitativo e transversal abrangeu os 853 municípios do estado de Minas Gerais. Todos os dados utilizados foram secundários, coletados e agrupados por regionais de planejamento. Como variáveis independentes, foram consideradas a cobertura de coleta de resíduos sólidos urbanos, cobertura de coleta seletiva e massa de resíduos sólidos urbanos, além de um indicador da qualidade da destinação final de resídulos, índices de desenvolvimento humano municipal e de Gini, renda mensal per capita e porcentagem de vulneráveis à pobreza. Os fatores potencialmente associados aos desfechos - incidências municipais de dengue, Chikungunya e Zika - foram selecionados inicialmente através de análises univariadas. Posteriormente, os modelos de regressão linear para as incidências de dengue, Chikungunya ou Zika foram gerados considerando os preditores selecionados pela análise univariada. Resultados. Não foi observada associação entre gestão de resíduos sólidos e incidência de Chikungunya e Zika. Por sua vez, a incidência de dengue associou-se à gestão de resíduos sólidos e apresentou relação inversa significativa com o percentual de vulneráveis à pobreza. Houve também associação direta o índice de Gini, sugerindo que quanto maiores os registros de incidência de dengue de 2007 a 2016, maiores os valores de Gini dos municípios - ou seja, maior a desigualdade social. A cobertura da coleta seletiva apresentou relação inversa e significativa com os casos de dengue, sugerindo que quanto menor a cobertura da coleta de seletiva, maiores foram os casos registrados de dengue. Conclusões. A gestão de resíduos sólidos pode influenciar os casos de dengue e, por isso, deve ser considerada nas ações de saúde pública.(AU)

ABSTRACT Objective. To investigate whether solid waste management and municipal socioeconomic indicators are associated with incidence rates of dengue, Zika, and Chikungunya in municipalities located in the state of Minas Gerais, Brazil. Methods. This exploratory, quantitative, cross-sectional study included all the 853 municipalities of Minas Gerais. Only secondary data were used, collected and grouped according to planning regions. Independent variables included regular urban solid waste collection, separated waste collection, and urban solid waste mass, in addition to a quality indicator of final waste disposal, municipal human development and Gini indices, monthly per capita income, and percentage of population vulnerable to poverty. The factors potentially associated with outcomes - municipal incidence of dengue, Chikungunya, and Zika - were initially selected by univariate analysis, followed by linear regression analysis for the incidence of dengue, Chikungunya, or Zika using the predictors selected through univariate analysis. Results. Solid waste management was not associated with incidence of Zika or Chikungunya. In turn, the incidence of dengue was associated with solid waste management and had a significant inverse association with percent population vulnerable to poverty. A direct association was also observed with Gini index, suggesting that the higher the incidence of dengue from 2007 to 2016, the higher the municipal Gini coefficient and thus social inequality. Selective waste collection was inversely and significantly correlated with dengue cases, suggesting that the lower the coverage by regular separated waste collection, the higher the number of dengue cases. Conclusions. Solid waste management may influence the occurrence of dengue cases, and therefore should be considered in the planning of public health actions.(AU)

RESUMEN Objetivo. Investigar si el manejo de los residuos sólidos y los indicadores socioeconómicos municipales están asociados con las tasas de incidencia de dengue, zika y chikunguña en los municipios del estado de Minas Gerais, Brasil. Métodos. Estudio exploratorio, cuantitativo y transversal que incluyó los 853 municipios de Minas Gerais. Sólo se utilizaron datos secundarios, recopilados y agrupados según las regiones de planificación. Las variables independientes incluyeron la recolección urbana regular de residuos sólidos, la recolección separada de residuos y la masa de residuos sólidos urbanos, además de un indicador de calidad de la eliminación final de desechos, el desarrollo humano municipal y los índices de Gini, el ingreso mensual per cápita y el porcentaje de población vulnerable a la pobreza. Los factores potencialmente asociados con los resultados -incidencia municipal de dengue, zika y chikunguña- se seleccionaron inicialmente mediante un análisis univariado, seguido de un análisis de regresión lineal para la incidencia del dengue, zika o chikunguña utilizando los predictores seleccionados mediante el análisis univariado. Resultados. El manejo de residuos sólidos no se asoció con la incidencia de zika o chikungunya. A su vez, la incidencia del dengue se asoció con un manejo sólido y tuvo una asociación inversa significativa con el porcentaje de población vulnerable a la pobreza. También se observó una asociación directa con el índice de Gini, lo que sugiere que cuanto mayor sea la incidencia del dengue entre 2007 y 2016, mayor será el coeficiente de Gini municipal y, por lo tanto, la desigualdad social. La recolección selectiva de residuos se correlacionó de manera significativa e inversa con los casos de dengue, lo que sugiere que cuanto menor sea la cobertura de la recolección regular selectiva de residuos, mayor será el número de casos de dengue. Conclusiones. La gestión de los residuos sólidos puede influir en la aparición de casos de dengue y, por lo tanto, debe considerarse en la planificación de las medidas de salud pública.(AU)