Coletanêa de Respostas Rápidas Turma 1 / Collection of Rapid Review Class 1

Coletânea dedicada aos estudos das respostas rápidas do Programa Educacional em Vigilância em Saúde no enfrentamento da COVID-19 e outras Doenças Virais (VigiEpidemia). Esse tema é de extrema relevância e atualidade em nosso contexto da saúde global e na resposta as emergências em saúde pública (ESP) de forma geral. As ESP, que englobam surtos e epidemias, desastres e desassistência à população, representam desafios complexos que exigem respostas ágeis e eficazes por parte das autoridades sanitárias, profissionais da saúde e comunidades como um todo. Até o momento, a pandemia de COVID-19 foi a maior ESP do Século XXI. Ela serviu como um lembrete doloroso da vulnerabilidade da humanidade diante da ameaça de doenças virais. Esta ESP, que teve resposta catastrófica em diversos momentos, evidenciou a importância do investimento em preparação, vigilância e resposta, destacando a necessidade de sistemas de vigilância robustos, colaboração internacional, Inteligência epidemiológica e comunicação transparente para mitigar o impacto devastador das doenças infecciosas na sociedade. As lições aprendidas com a pandemia de COVID-19 são vastas e multifacetadas. A importância da pesquisa, da educação em saúde e do desenvolvimento de vacinas foi evidenciada como uma prioridade crucial na proteção da saúde pública mundial. O investimento em pesquisas e em cursos para formação de profissionais que possam estar atentos as mudanças nos padrões e comportamentos das doenças infecciosas, além de atuar na resposta rápida quando necessário, é fundamental para estarmos preparados para as futuras pandemias. A vacinação, por exemplo, sempre foi uma das ferramentas mais poderosas para evitar surtos e epidemias e, durante a pandemia de COVID-19, ajudou a controlar os óbitos pela doença e possibilitou que voltássemos a ter uma vida normal. Além da vacina contra COVID-19, as vacinas de influenza e dengue também são exemplos notáveis de avanços científicos que desempenham um papel fundamental na prevenção de futuras ESP. Ao explorar os diversos aspectos da resposta, monitoramento e controle de surtos, epidemias e pandemias, esta coletânea visa fornecer uma compreensão abrangente dos desafios enfrentados, das melhores práticas e das estratégias eficazes para mitigar os impactos adversos desses eventos. Espera-se que este trabalho não apenas informe e eduque, mas também inspire ações concretas para fortalecer a recuperação e resiliência dos sistemas de saúde e proteger o bem-estar das comunidades mais vulneráveis do nosso pais.

A collection dedicated to the study of rapid responses by the Educational Program in Health Surveillance in addressing COVID-19 and other Viral Diseases (VigiEpidemia). This theme is of utmost relevance and timeliness in our context of global health and in responding to public health emergencies (PHE) in general. PHEs, which encompass outbreaks and epidemics, disasters, and neglect of the population, represent complex challenges that require swift and effective responses from health authorities, healthcare professionals, and communities as a whole. To date, the COVID-19 pandemic has been the largest PHE of the 21st century. It served as a painful reminder of humanity's vulnerability in the face of viral disease threats. This PHE, which had catastrophic responses at various times, highlighted the importance of investing in preparedness, surveillance, and response, underscoring the need for robust surveillance systems, international collaboration, epidemiological intelligence, and transparent communication to mitigate the devastating impact of infectious diseases on society. The lessons learned from the COVID-19 pandemic are vast and multifaceted. The importance of research, health education, and vaccine development was highlighted as a crucial priority in protecting global public health. Investing in research and training courses to prepare professionals who can be attentive to changes in the patterns and behaviors of infectious diseases and act quickly when needed is essential to be prepared for future pandemics. Vaccination, for example, has always been one of the most powerful tools to prevent outbreaks and epidemics, and during the COVID-19 pandemic, it helped control disease-related deaths and allowed us to return to a normal life. In addition to the COVID-19 vaccine, influenza and dengue vaccines are also notable examples of scientific advancements that play a key role in preventing future PHEs. By exploring the various aspects of response, monitoring, and control of outbreaks, epidemics, and pandemics, this collection aims to provide a comprehensive understanding of the challenges faced, best practices, and effective strategies to mitigate the adverse impacts of these events. It is hoped that this work will not only inform and educate but also inspire concrete actions to strengthen the recovery and resilience of health systems and protect the well-being of the most vulnerable communities in our country.

Interleukin 27, like interferons, activates JAK-STAT signaling and promotes pro-inflammatory and antiviral states that interfere with dengue and chikungunya viruses replication in human macrophages.

Interferons (IFNs) are a family of cytokines that activate the JAK-STAT signaling pathway to induce an antiviral state in cells. Interleukin 27 (IL-27) is a member of the IL-6 and/or IL-12 family that elicits both pro- and anti-inflammatory responses. Recent studies have reported that IL-27 also induces a robust antiviral response against diverse viruses, both in vitro and in vivo, suggesting that IFNs and IL-27 share many similarities at the functional level. However, it is still unknown how similar or different IFN- and IL-27-dependent signaling pathways are. To address this question, we conducted a comparative analysis of the transcriptomic profiles of human monocyte-derived macrophages (MDMs) exposed to IL-27 and those exposed to recombinant human IFN-α, IFN-γ, and IFN-λ. We utilized bioinformatics approaches to identify common differentially expressed genes between the different transcriptomes. To verify the accuracy of this approach, we used RT-qPCR, ELISA, flow cytometry, and microarrays data. We found that IFNs and IL-27 induce transcriptional changes in several genes, including those involved in JAK-STAT signaling, and induce shared pro-inflammatory and antiviral pathways in MDMs, leading to the common and unique expression of inflammatory factors and IFN-stimulated genes (ISGs)Importantly, the ability of IL-27 to induce those responses is independent of IFN induction and cellular lineage. Additionally, functional analysis demonstrated that like IFNs, IL-27-mediated response reduced chikungunya and dengue viruses replication in MDMs. In summary, IL-27 exhibits properties similar to those of all three types of human IFN, including the ability to stimulate a protective antiviral response. Given this similarity, we propose that IL-27 could be classified as a distinct type of IFN, possibly categorized as IFN-pi (IFN-π), the type V IFN (IFN-V).

Chikungunya virus infection in human microglial C20 cells induces mitochondria-mediated apoptosis.

Introduction: Chikungunya virus (CHIKV) infection is associated with acute clinical manifestations and chronic joint inflammation. CHIKV has emerged as a significant causative agent of central nervous system (CNS) complications, including encephalitis and related sequelae. Microglial cells, crucial for immune responses and tissue repair in the CNS, play a vital role in the host response to viral infections, with their activation potentially leading to either protection or pathology. In this study, the infection biology of CHIKV in the C20 human microglial cell line was investigated. Methods: The permissiveness of C20 cells to CHIKV infection was assessed, and viral replication kinetics were compared to Vero E6 cells. Cytopathic effects of CHIKV infection on C20 cells were examined, along with ultrastructural changes using transmission electron microscopy. Additionally, apoptosis induction, mitochondrial membrane potential, and alterations in cell surface marker expression were evaluated by flow cytometry. Results: CHIKV infection demonstrated permissiveness in C20 cells, similar to Vero cells, resulting in robust viral replication and cytopathic effects. Ultrastructural analysis revealed viral replication, mature virion formation, and distinctive cytoplasmic and nuclear changes in infected C20 cells. CHIKV infection induced significant apoptosis in C20 cells, accompanied by mitochondrial membrane depolarization and altered expression of cell surface markers such as CD11c, CD14, and HLA-DR. Notably, decreased CD14 expression was observed in CHIKV-infected C20 cells. Discussion: The study findings suggest that CHIKV infection induces apoptosis in C20 microglial cells via the mitochondrial pathway, with significant alterations in cell surface marker expression, particularly CD14 that is linked with apoptosis induction. These observations provide valuable insights into the role of human microglial cells in the host response to CHIKV infection and contribute to the knowledge on the neuropathogenesis of this virus.

Spatial distribution of arboviruses and its association with a social development index and the waste disposal in São Luís, state of Maranhão, Brazil, 2015 to 2019.

OBJECTIVE: To detect spatial and spatiotemporal clusters of urban arboviruses and to investigate whether the social development index (SDI) and irregular waste disposal are related to the coefficient of urban arboviruses detection in São Luís, state of Maranhão, Brazil. METHODS: The confirmed cases of Dengue, Zika and Chikungunya in São Luís, from 2015 to 2019, were georeferenced to the census tract of residence. The Bayesian Conditional Autoregressive regression model was used to identify the association between SDI and irregular waste disposal sites and the coefficient of urban arboviruses detection. RESULTS: The spatial pattern of arboviruses pointed to the predominance of a low-incidence cluster, except 2016. For the years 2015, 2016, 2017, and 2019, an increase of one unit of waste disposal site increased the coefficient of arboviruses detection in 1.25, 1.09, 1.23, and 1.13 cases of arboviruses per 100 thousand inhabitants, respectively. The SDI was not associated with the coefficient of arboviruses detection. CONCLUSION: In São Luís, spatiotemporal risk clusters for the occurrence of arboviruses and a positive association between the coefficient of arbovirus detection and sites of irregular waste disposal were identified.

Comparative analysis of midgut bacterial communities in Chikungunya virus-infected and non-infected Aedes aegypti Thai laboratory strain mosquitoes.

Chikungunya virus (CHIKV) poses a significant global health threat, re-emerging as a mosquito-transmitted pathogen that caused high fever, rash, and severe arthralgia. In Thailand, a notable CHIKV outbreak in 2019-2020 affected approximately 20,000 cases across 60 provinces, underscoring the need for effective mosquito control protocols. Previous studies have highlighted the role of midgut bacteria in the interaction between mosquito vectors and pathogen infections, demonstrating their ability to protect the insect from invading pathogens. However, research on the midgut bacteria of Aedes (Ae.) aegypti, the primary vector for CHIKV in Thailand remains limited. This study aims to characterize the bacterial communities in laboratory strains of Ae. aegypti, both infected and non-infected with CHIKV. Female mosquitoes from a laboratory strain of Ae. aegypti were exposed to a CHIKV-infected blood meal through membrane feeding, while the control group received a non-infected blood meal. At 7 days post-infection (dpi), mosquito midguts were dissected for 16S rRNA gene sequencing to identify midgut bacteria, and CHIKV presence was confirmed by E1-nested RT-PCR using mosquito carcasses. The study aimed to compare the bacterial communities between CHIKV-infected and non-infected groups. The analysis included 12 midgut bacterial samples, divided into three groups: CHIKV-infected (exposed and infected), non-infected (exposed but not infected), and non-exposed (negative control). Alpha diversity indices and Bray-Curtis dissimilarity matrix revealed significant differences in bacterial profiles among the three groups. The infected group exhibited an increased abundance of bacteria genus Gluconobacter, while Asaia was prevalent in both non-infected and negative control groups. Chryseobacterium was prominent in the negative control group. These findings highlight potential alterations in the distribution and abundance of gut microbiomes in response to CHIKV infection status. This study provides valuable insights into the dynamic relationship between midgut bacteria and CHIKV, underscoring the potential for alterations in bacterial composition depending on infection status. Understanding the relationships between mosquitoes and their microbiota holds promise for developing new methods and tools to enhance existing strategies for disease prevention and control. This research advances our understanding of the circulating bacterial composition, opening possibilities for new approaches in combating mosquito-borne diseases.

Exposure to Zika and chikungunya viruses impacts aspects of the vectorial capacity of Aedes aegypti and Culex quinquefasciatus.

Zika (ZIKV) and chikungunya (CHIKV) are arboviruses that cause infections in humans and can cause clinical complications, representing a worldwide public health problem. Aedes aegypti is the primary vector of these pathogens and Culex quinquefasciatus may be a potential ZIKV vector. This study aimed to evaluate fecundity, fertility, survival, longevity, and blood feeding activity in Ae. aegypti after exposure to ZIKV and CHIKV and, in Cx. quinquefasciatus exposed to ZIKV. Three colonies were evaluated: AeCamp (Ae. aegypti-field), RecL (Ae. aegypti-laboratory) and CqSLab (Cx. quinquefasciatus-laboratory). Seven to 10 days-old females from these colonies were exposed to artificial blood feeding with CHIKV or ZIKV. CHIKV caused reduction in fecundity and fertility in AeCamp and reduction in survival and fertility in RecL. ZIKV impacted survival in RecL, fertility in AeCamp and, fecundity and fertility in CqSLab. Both viruses had no effect on blood feeding activity. These results show that CHIKV produces a higher biological cost in Ae. aegypti, compared to ZIKV, and ZIKV differently alters the biological performance in colonies of Ae. aegypti and Cx. quinquefasciatus. These results provide a better understanding over the processes of virus-vector interaction and can shed light on the complexity of arbovirus transmission.

A Highly Sensitive Molecular Technique for RNA Virus Detection.

Zika (ZIKV) and Chikungunya (CHIKV) viruses are mosquito-transmitted infections, or vector-borne pathogens, that emerged a few years ago. Reliable diagnostic tools for ZIKV and CHIKV-inexpensive, multiplexed, rapid, highly sensitive, and specific point-of-care (POC) systems-are vital for appropriate risk management and therapy. We recently studied a detection system with great success in Mexico (Villahermosa, state of Tabasco), working with human sera from patients infected with those viruses. The research conducted in Mexico validated the efficacy of a novel two-step rapid isothermal amplification technique (RAMP). This approach, which encompasses recombinase polymerase amplification (RPA) followed by loop-mediated isothermal amplification (LAMP), had been previously established in the lab using lab-derived Zika (ZIKV) and Chikungunya (CHIKV) viruses. Crucially, our findings confirmed that this technique is also effective when applied to human sera samples collected from locally infected individuals in Mexico.

In vivo rescue of arboviruses directly from subgenomic DNA fragments.

Reverse genetic systems are mainly used to rescue recombinant viral strains in cell culture. These tools have also been used to generate, by inoculating infectious clones, viral strains directly in living animals. We previously developed the "Infectious Subgenomic Amplicons" (ISA) method, which enables the rescue of single-stranded positive sense RNA viruses in vitro by transfecting overlapping subgenomic DNA fragments. Here, we provide proof-of-concept for direct in vivo generation of infectious particles following the inoculation of subgenomic amplicons. First, we rescued a strain of tick-borne encephalitis virus in mice to transpose the ISA method in vivo. Subgenomic DNA fragments were amplified using a 3-fragment reverse genetics system and inoculated intramuscularly. Almost all animals were infected when quantities of DNA inoculated were at least 20 µg. We then optimized our procedure in order to increase the animal infection rate. This was achieved by adding an electroporation step and/or using a simplified 2- fragment reverse genetics system. Under optimal conditions, a large majority of animals were infected with doses of 20 ng of DNA. Finally, we demonstrated the versatility of this method by applying it to Japanese encephalitis and Chikungunya viruses. This method provides an efficient strategy for in vivo rescue of arboviruses. Furthermore, in the context of the development of DNA-launched live attenuated vaccines, this new approach may facilitate the generation of attenuated strains in vivo. It also enables to deliver a substance free of any vector DNA, which seems to be an important criterion for the development of human vaccines.

The rising global economic costs of invasive Aedes mosquitoes and Aedes-borne diseases.

Invasive Aedes aegypti and Aedes albopictus mosquitoes transmit viruses such as dengue, chikungunya and Zika, posing a huge public health burden as well as having a less well understood economic impact. We present a comprehensive, global-scale synthesis of studies reporting these economic costs, spanning 166 countries and territories over 45 years. The minimum cumulative reported cost estimate expressed in 2022 US$ was 94.7 billion, although this figure reflects considerable underreporting and underestimation. The analysis suggests a 14-fold increase in costs, with an average annual expenditure of US$ 3.1 billion, and a maximum of US$ 20.3 billion in 2013. Damage and losses were an order of magnitude higher than investment in management, with only a modest portion allocated to prevention. Effective control measures are urgently needed to safeguard global health and well-being, and to reduce the economic burden on human societies. This study fills a critical gap by addressing the increasing economic costs of Aedes and Aedes-borne diseases and offers insights to inform evidence-based policy.

Detection of musculoskeletal inflammatory lesions in patients with chronic chikungunya infection using 3T whole-body magnetic resonance imaging.

BACKGROUND: Musculoskeletal inflammatory lesions in chronic Chikungunya virus (CHIKV) infection have not been thoroughly assessed using whole-body magnetic resonance imaging (WBMRI). This study aimed to determine the prevalence of these lesions in such patients. METHODS: From September 2018 to February 2019, patients with positive Chikungunya-specific serology (Immunoglobulin M/Immunoglobulin G anti-CHIKV), with a history of polyarthralgia for > 6 months prior to MRI with no pre-existing rheumatic disorders, underwent 3T WBMRI and localized MRI. The evaluation focused on musculoskeletal inflammatory lesions correlated with chronic CHIKV infection. Pain levels were assessed using a visual analogue scale on the same day as WBMRI. RESULTS: The study included 86 patients of whom 26 met the inclusion criteria. All patients reported pain and most (92.3%) categorized it as moderate or severe. The most common finding across joints was effusion, particularly in the tibiotalar joint (57.7%) and bursitis, with the retrocalcaneal bursa most affected (48.0%). Tenosynovitis was prevalent in the flexor compartment of the hands (44.2%), while Kager fat pad and soleus edema were also observed. Bone marrow edema-like signals were frequently seen in the sacroiliac joints (19.2%). Most WBMRI findings were classified as mild. CONCLUSIONS: This study represents the first utilization of 3T WBMRI to assess musculoskeletal inflammatory disorders in chronic CHIKV infection. The aim was to identify the most affected joints and prevalent lesions, providing valuable insights for future research and clinical management of this condition regarding understanding disease pathophysiology, developing targeted treatment strategies, and using advanced imaging techniques in the assessment of musculoskeletal manifestations.

Unlocking the antiviral potential of rosmarinic acid against chikungunya virus via IL-17 signaling pathway.

Background: The Chikungunya virus is an Alphavirus that belongs to the Togaviridae family and is primarily transmitted by mosquitoes. It causes acute infection characterized by fever, headache, and arthralgia. Some patients also experience persistent chronic osteoarthritis-like symptoms. Dedicated antiviral treatments are currently unavailable for CHIKV. This study aims to explore the potential anti-CHIKV effect of rosmarinic acid using network pharmacology. Methods: This study employed network pharmacology to predict and verify the molecular targets and pathways associated with ROSA in the context of CHIKV. The analysis outcomes were further validated using molecular docking and in vitro experiments. Results: The analysis of CHIKV targets using the Kyoto Encyclopedia of Genes and Genomes and MCODE identified IL-17 as an important pathogenic pathway in CHIKV infection. Among the 30 targets of ROSA against CHIKV, nearly half were found to be involved in the IL-17 signaling pathway. This suggests that ROSA may help the host in resisting CHIKV invasion by modulating this pathway. Molecular docking validation results showed that ROSA can stably bind to 10 core targets out of the 30 identified targets. In an in vitro CHIKV infection model developed using 293T cells, treatment with 60 µM ROSA significantly improved the survival rate of infected cells, inhibited 50% CHIKV proliferation after CHIKV infection, and reduced the expression of TNF-α in the IL-17 signaling pathway. Conclusion: This study provides the first confirmation of the efficacy of ROSA in suppressing CHIKV infection through the IL-17 signaling pathway. The findings warrant further investigation to facilitate the development of ROSA as a potential treatment for CHIKV infection.

Molecular identification and phylogenetic analysis of chikungunya virus among dengue-negative patients in Kolkata, India.

Dengue and chikungunya are co-circulating vector-borne diseases that share a significant number of clinical symptoms. To identify variables to aid physicians in making rapid and effective diagnostic decisions, we performed molecular diagnosis of the chikungunya virus and examined the clinical manifestations of chikungunya cases to identify the prevalence among dengue-negative individuals in Kolkata. Dengue suspected patients' samples were collected during January 2020-December 2021 and Enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) methods have been performed to confirm the prevalence of chikungunya infection among dengue-negative patients. By performing phylogenetic analysis, comparing clinical classifications, identifying disease aetiology using clinical and laboratory factors, and evaluating the time course of several clinical variables, we have evaluated the clinical manifestations linked to dengue and chikungunya virus infections. Chikungunya infection was found in 15.1% and 6.3% of the 635 dengue-negative patients, as determined by ELISA and RT-PCR, respectively. Arthritis and myalgia were more common in chikungunya-infected patients at the time of hospital admission while conjunctivitis, photosensitivity, arthralgia, Anorexia, fatigue, retro-orbital pain, vomiting, dermatitis, or swollen glands were significantly presented as an overlapping symptom. Although dengue and chikungunya infections have significant clinical overlap, basic clinical and laboratory criteria can predict these diseases at presentation for proper management. Effective management enables doctors to treat and care for patients properly and contributes to the development of control measures for these infections in a medical setting.

Design, development, and validation of multi-epitope proteins for serological diagnosis of Zika virus infections and discrimination from dengue virus seropositivity.

Zika virus (ZIKV), an arbovirus from the Flaviviridae family, is the causative agent of Zika fever, a mild and frequent oligosymptomatic disease in humans. Nonetheless, on rare occasions, ZIKV infection can be associated with Guillain-Barré Syndrome (GBS), and severe congenital complications, such as microcephaly. The oligosymptomatic disease, however, presents symptoms that are quite similar to those observed in infections caused by other frequent co-circulating arboviruses, including dengue virus (DENV). Moreover, the antigenic similarity between ZIKV and DENV, and even with other members of the Flaviviridae family, complicates serological testing due to the high cross-reactivity of antibodies. Here, we designed, produced in a prokaryotic expression system, and purified three multiepitope proteins (ZIKV-1, ZIKV-2, and ZIKV-3) for differential diagnosis of Zika. The proteins were evaluated as antigens in ELISA tests for the detection of anti-ZIKV IgG using ZIKV- and DENV-positive human sera. The recombinant proteins were able to bind and detect anti-ZIKV antibodies without cross-reactivity with DENV-positive sera and showed no reactivity with Chikungunya virus (CHIKV)- positive sera. ZIKV-1, ZIKV-2, and ZIKV-3 proteins presented 81.6%, 95%, and 66% sensitivity and 97%, 96%, and 84% specificity, respectively. Our results demonstrate the potential of the designed and expressed antigens in the development of specific diagnostic tests for the detection of IgG antibodies against ZIKV, especially in regions with the circulation of multiple arboviruses.

[Dengue, chikungunya and the mosquito vector at the Southern limit of distribution during the 2023 epidemic, Argentina]. / Dengue, chikungunya y el mosquito vector en el límite sur de distribución durante la epidemia 2023, Argentina.

OBJECTIVES: To monitor the oviposition activity of the mosquito Aedes aegypti and of dengue and chikungunya cases in four localities of temperate Argentina, during the 2023 epidemic. METHODS: During the summer and autumn of 2023, the oviposition activity of the mosquito vector was monitored weekly using ovitraps, and the arrival of cases with dengue or chikungunya in Tandil, Olavarría, Bahía Blanca and Laprida were registered. RESULTS: Monthly variations of the percentage of positive traps were similar in the first three locations; in Laprida the mosquito was not detected. On the contrary, a significant difference was observed in the percentage of total traps that ever tested positive in each locality, being higher in Olavarría (83.3%) than in Bahía Blanca (68.6%) and Tandil (48.7%). Regarding diseases, 18 imported cases of dengue and 3 of chikungunya were registered. In addition, the first autochthonous case of dengue in the region was recorded, being the southernmost until known. CONCLUSION: It is essential to raise awareness and train the members of the health systems of the new regions exposed to Ae. aegypti for early detection of cases, and to the general population to enhance prevention actions.

OBJETIVOS: Monitorear la actividad de oviposición del mosquito Aedes aegypti y de casos de dengue y chikungunya en cuatro localidades de Argentina templada, durante la epidemia del 2023. Métodos: Durante el verano y otoño del 2023, se monitoreó semanalmente mediante ovitrampas la actividad de oviposición del mosquito vector, y se registró el arribo de casos con dengue o chikungunya a Tandil, Olavarría, Bahía Blanca y Laprida. RESULTADOS: La variación mensual del porcentaje de trampas positivas fue similar en las tres primeras localidades; en Laprida no se detectó el mosquito. Por el contrario, se observó una diferencia significativa del porcentaje de trampas que alguna vez resultó positiva en cada localidad, siendo mayor en Olavarría (83%), que en Bahía Blanca (67%) y Tandil (49%). Respecto a las enfermedades, se registraron 18 casos importados de dengue y 3 de chikungunya. Además, se registró el primer caso autóctono de dengue en la región, siendo el más austral hasta el momento. Conclusión: Es imprescindible sensibilizar y capacitar a los integrantes de los sistemas de salud de las nuevas regiones expuestas al Ae. aegypti para la detección temprana de casos, y a la población en general para potenciar las acciones de prevención.

A case series of post-infectious chikungunya myeloradiculoneuropathy.

Chikungunya fever is an arboviral illness caused by chikungunya virus (CHIKV) and transmitted by the bite of Aedes aegypti and Aedes albopictus. It is an RNA virus belonging to the genus Alphavirus and family Togaviridae. We present a case series of three patients with chikungunya illness developing para/post-infectious myeloradiculoneuropathy.These patients developed neurological symptoms in the form of bilateral lower limb weakness with sensory and bowel involvement after the recovery from the initial acute episode of chikungunya fever. Clinical examination findings suggested myeloradiculoneuropathy with normal Magnetic Resonance Imaging of the Spine, with the nerve conduction study showing sensorimotor axonal polyneuropathy. All the patients were treated with 1 g of methylprednisolone once a day for five days, and case 2 was given intravenous immunoglobulin also. In the follow-up, cases 1 and 2 showed complete recovery without recurrence, and case 3 did not show improvement at one month.

Bone erosions and joint damage caused by chikungunya virus: a systematic review.

BACKGROUND: Chikungunya fever is an emerging global infection transmitted by Aedes mosquitoes that manifests as an acute febrile illness with joint pain and can lead to chronic arthritis. The mechanism underlying chronic joint damage remains unclear; however, chronic chikungunya arthritis shares similarities with rheumatoid arthritis. Disease-modifying antirheumatic drugs have revolutionized rheumatoid arthritis treatment by preventing joint damage. However, the role of these therapies in chronic chikungunya arthritis has not been determined. We conducted a systematic review to evaluate the burden of joint structural damage in chronic chikungunya arthritis to help to define the role of disease-modifying therapy in this disease. METHODS: This systematic review included retrospective and prospective studies, trials, and case reports evaluating joint damage caused by chikungunya virus. Various databases were searched without any date or language restrictions. Study selection was conducted independently by two researchers, and data were extracted from the articles selected. RESULTS: A total of 108 studies were initially evaluated, with 8 meeting the inclusion criteria. Longitudinal studies have reported persistent joint pain from chikungunya infection and the progression of radiographic joint damage up to 13 years post-infection. Joint imaging revealed synovial inflammation, bone erosion, and cartilage destruction in patients with chronic chikungunya arthritis. CONCLUSIONS: Few studies have addressed chikungunya-induced joint damage, limiting our understanding of chronic chikungunya arthritis. Nevertheless, chronic chikungunya arthritis has similarities to rheumatoid arthritis. The success of early disease-modifying antirheumatic drug therapy in rheumatoid arthritis underscores the need for comprehensive research on its role in chikungunya arthritis.

The High Capacity of Brazilian Aedes aegypti Populations to Transmit a Locally Circulating Lineage of Chikungunya Virus.

The incidence of chikungunya has dramatically surged worldwide in recent decades, imposing an expanding burden on public health. In recent years, South America, particularly Brazil, has experienced outbreaks that have ravaged populations following the rapid dissemination of the chikungunya virus (CHIKV), which was first detected in 2014. The primary vector for CHIKV transmission is the urban mosquito species Aedes aegypti, which is highly prevalent throughout Brazil. However, the impact of the locally circulating CHIKV genotypes and specific combinations of local mosquito populations on vector competence remains unexplored. Here, we experimentally analyzed and compared the infectivity and transmissibility of the CHIKV-ECSA lineage recently isolated in Brazil among four Ae. aegypti populations collected from different regions of the country. When exposed to CHIKV-infected AG129 mice for blood feeding, all the mosquito populations displayed high infection rates and dissemination efficiency. Furthermore, we observed that all the populations were highly efficient in transmitting CHIKV to a vertebrate host (naïve AG129 mice) as early as eight days post-infection. These results demonstrate the high capacity of Brazilian Ae. aegypti populations to transmit the locally circulating CHIKV-ECSA lineage. This observation could help to explain the high prevalence of the CHIKV-ECSA lineage over the Asian lineage, which was also detected in Brazil in 2014. However, further studies comparing both lineages are necessary to gain a better understanding of the vector's importance in the epidemiology of CHIKV in the Americas.

U-CAN-seq: A Universal Competition Assay by Nanopore Sequencing.

RNA viruses quickly evolve subtle genotypic changes that can have major impacts on viral fitness and host range, with potential consequences for human health. It is therefore important to understand the evolutionary fitness of novel viral variants relative to well-studied genotypes of epidemic viruses. Competition assays are an effective and rigorous system with which to assess the relative fitness of viral genotypes. However, it is challenging to quickly and cheaply distinguish and quantify fitness differences between very similar viral genotypes. Here, we describe a protocol for using reverse transcription PCR in combination with commercial nanopore sequencing services to perform competition assays on untagged RNA viruses. Our assay, called the Universal Competition Assay by Nanopore Sequencing (U-CAN-seq), is relatively cheap and highly sensitive. We used a well-studied N24A mutation in the chikungunya virus (CHIKV) nsp3 gene to confirm that we could detect a competitive disadvantage using U-CAN-seq. We also used this approach to show that mutations to the CHIKV 5' conserved sequence element that disrupt sequence but not structure did not affect the fitness of CHIKV. However, similar mutations to an adjacent CHIKV stem loop (SL3) did cause a fitness disadvantage compared to wild-type CHIKV, suggesting that structure-independent, primary sequence determinants in this loop play an important role in CHIKV biology. Our novel findings illustrate the utility of the U-CAN-seq competition assay.

Live-attenuated CHIKV vaccine with rearranged genome replicates in vitro and induces immune response in mice.

Chikungunya fever virus (CHIKV) is a mosquito-borne alphavirus that causes wide-spread human infections and epidemics in Asia, Africa and recently, in the Americas. CHIKV is considered a priority pathogen by CEPI and WHO. Despite recent approval of a live-attenuated CHIKV vaccine, development of additional vaccines is warranted due to the worldwide outbreaks of CHIKV. Previously, we developed immunization DNA (iDNA) plasmid capable of launching live-attenuated CHIKV vaccine in vivo. Here we report the use of CHIKV iDNA plasmid to prepare a novel, live-attenuated CHIKV vaccine V5040 with rearranged RNA genome. In V5040, genomic RNA was rearranged to encode capsid gene downstream from the glycoprotein genes. Attenuated mutations derived from experimental CHIKV 181/25 vaccine were also engineered into E2 gene of V5040. The DNA copy of rearranged CHIKV genomic RNA with attenuated mutations was cloned into iDNA plasmid pMG5040 downstream from the CMV promoter. After transfection in vitro, pMG5040 launched replication of V5040 virus with rearranged genome and attenuating E2 mutations. Furthermore, V5040 virus was evaluated in experimental murine models for general safety and immunogenicity. Vaccination with V5040 virus subcutaneously resulted in elicitation of CHIKV-specific, virus-neutralizing antibodies. The results warrant further evaluation of V5040 virus with rearranged genome as a novel live-attenuated vaccine for CHIKV.