Showing papers on "Dengue virus published in 2022"

Identification of acridinedione scaffolds as potential inhibitor of DENV‐2 C protein: An in silico strategy to combat dengue

Abstract: Dengue is a prominent viral disease transmitted by mosquitoes to humans that affects mainly tropical and subtropical countries worldwide. The global spread of dengue virus (DENV) is mainly occurred by Aedes aegypti and Aedes albopictus mosquitoes. The dengue virus serotypes‐2 (DENV‐2) is a widely prevalent serotype of DENV, that causes the hemorrhagic fever and bleeding in the mucosa, which can be fatal. In the life cycle of DENV‐2, a structural capsid (DENV‐2 C) protein forms the nucleocapsid assembly and bind to the viral progeny RNA. For DENV‐2 maturation, the nucleocapsid is a vital component. We used virtual ligand screening to filter out the best in‐house synthesized acridinedione analogs (DSPD molecules) that could efficiently bind to DENV‐2 C protein. The molecular docking and dynamics simulations studies were performed to analyze the effect of DSPD molecules on DENV‐2 C protein after binding. Our findings showed that DSPD molecules strongly interacted with DENV‐2 C protein, as evident from molecular interactions and several time‐dependent molecular dynamics‐driven analyses. Moreover, this study was also supported by the thermodynamic binding free energy and steered molecular dynamics simulations. Therefore, we intend to suggest that the DSPD3 molecule could be used as a potential therapeutic molecule against dengue complications as compared to the cocrystallized inhibitor ST‐148. However, further studies are required to demonstrate the ability of DSPD3 to induce DENV‐2 C tetramer formation.

Dengue: A Growing Problem With New Interventions.

Abstract: Dengue is the disease caused by 1 of 3 distinct, but closely related dengue viruses (DENV-1-4) that are transmitted by Aedes spp. mosquito vectors. It is the most common arboviral disease worldwide, with the greatest burden in tropical and sub-tropical regions. In the absence of effective prevention and control measures, dengue is projected to increase in both disease burden and geographic range. Given its increasing importance as an etiology of fever in the returning traveler or the possibility of local transmission in regions in the United States with competent vectors, as well as the risk for large outbreaks in endemic US territories and associated states, clinicians should understand its clinical presentation and be familiar with appropriate testing, triage, and management of patients with dengue. Control and prevention efforts reached a milestone in June 2021 when the Advisory Committee on Immunization Practices (ACIP) recommended Dengvaxia for routine use in children aged 9 to 16 years living in endemic areas with laboratory confirmation of previous dengue virus infection. Dengvaxia is the first vaccine against dengue to be recommended for use in the United States and one of the first to require laboratory testing of potential recipients to be eligible for vaccination. In this review, we outline dengue pathogenesis, epidemiology, and key clinical features for front-line clinicians evaluating patients presenting with dengue. We also provide a summary of Dengvaxia efficacy, safety, and considerations for use as well as an overview of other potential new tools to control and prevent the growing threat of dengue .

Vaccine-Associated Enhanced Disease and Pathogenic Human Coronaviruses

Abstract: Vaccine-associated enhanced disease (VAED) is a difficult phenomenon to define and can be confused with vaccine failure. Using studies on respiratory syncytial virus (RSV) vaccination and dengue virus infection, we highlight known and theoretical mechanisms of VAED, including antibody-dependent enhancement (ADE), antibody-enhanced disease (AED) and Th2-mediated pathology. We also critically review the literature surrounding this phenomenon in pathogenic human coronaviruses, including MERS-CoV, SARS-CoV-1 and SARS-CoV-2. Poor quality histopathological data and a lack of consistency in defining severe pathology and VAED in preclinical studies of MERS-CoV and SARS-CoV-1 vaccines in particular make it difficult to interrogate potential cases of VAED. Fortuitously, there have been only few reports of mild VAED in SARS-CoV-2 vaccination in preclinical models and no observations in their clinical use. We describe the problem areas and discuss methods to improve the characterisation of VAED in the future.

Antibody Fc characteristics and effector functions correlate with protection from symptomatic dengue virus type 3 infection

Abstract: Preexisting cross-reactive antibodies have been implicated in both protection and pathogenesis during subsequent infections with different dengue virus (DENV) serotypes (DENV1-4). Nonetheless, humoral immune correlates and mechanisms of protection have remained elusive. Using a systems serology approach to evaluate humoral responses, we profiled plasma collected before inapparent or symptomatic secondary DENV3 infection from our pediatric cohort in Nicaragua. Children protected from symptomatic infections had more anti-envelope (E) and anti–nonstructural protein 1 (NS1) total immunoglobulin G (IgG), IgG4, and greater Fc effector functions than those with symptoms. Fc effector functions were also associated with protection from hemorrhagic manifestations in the pre-symptomatic group. Furthermore, in vitro virological assays using these plasma samples revealed that protection mediated by antibody-dependent complement deposition was associated with both lysis of virions and DENV-infected cells. These data suggest that E- and NS1-specific Fc functions may serve as correlates of protection, which can be potentially applied toward the design and evaluation of dengue vaccines. Description Antibody Fc characteristics and effector functions correlate with protection from symptomatic DENV3 infection. Dengue immune defenses Cross-reactive antibodies against different dengue virus serotypes (DENV1-4) can have either protective or pathogenic effects depending on immune responses to the secondary infection serotype. Dias et al. used systems serology to profile DENV-specific antibody responses of previously infected children in a Nicaraguan cohort study to determine correlates of protection against secondary DENV3 infection. Children protected against secondary symptomatic infection had higher titers of total IgG and IgG4 specific to envelope (E) protein and nonstructural protein 1 (NS1) and had antibodies with greater Fc effector functions relative to symptomatic children. Plasma anti-E and anti-NS1 polyclonal antibodies were associated with protection via Fc-driven complement deposition and lysis of DENV virions or infected cells in vitro. Together, these observations indicate that E- and NS1-specific Abs may have Fc effector functions that serve as correlates of protection against DENV.

Impact of temperature on dengue and chikungunya transmission by the mosquito Aedes albopictus

Abstract: Abstract The mosquito Aedes albopictus is an invasive species first detected in Europe in Albania in 1979, and now established in 28 European countries. Temperature is a limiting factor in mosquito activities and in the transmission of associated arboviruses namely chikungunya (CHIKV) and dengue (DENV). Since 2007, local transmissions of CHIKV and DENV have been reported in mainland Europe, mainly in South Europe. Thus, the critical question is how far north transmission could occur. In this context, the Albanian infestation by Ae. albopictus is of interest because the species is present up to 1200 m of altitude; this allows using altitude as a proxy for latitude. Here we show that Ae. albopictus can transmit CHIKV at 28 °C as well as 20 °C, however, the transmission of DENV is only observed at 28 °C. We conclude that if temperature is the key environmental factor limiting transmission, then transmission of CHIKV, but not DENV is feasible in much of Europe.

Seeking heterocyclic scaffolds as antivirals against dengue virus

Abstract: Dengue is one of the most typical viral infection categorized in the Neglected Tropical Diseases (NTDs). It is transmitted via the female Aedes aegypti mosquito to humans and majorly puts risk to the lives of more than half of the world. Recent advancements in medicinal chemistry have led to the design and development of numerous potential heterocyclic scaffolds as antiviral drug candidates for the inhibition of the dengue virus (DENV). Thus, in this review, we have discussed the significance of inhibitory and antiviral activities of nitrogen, oxygen, and mixed (nitrogen - sulfur and nitrogen-oxygen) heterocyclic scaffolds that are published in the last seven years (2016–2022). Furthermore, we have also discussed the probable mechanisms of action and the diverse structure-activity relationships (SARs) of the heterocyclic scaffolds. In addition, this review has elaborately outlined the mechanism of viral infection and the life cycle of DENV in the host cells. The wide set of heterocycles and their SARs will aid in the development of pharmaceuticals that will allow the researchers to synthesize the promising anti-dengue drug candidate in the future. • Dengue is a critical viral infection under Neglected Tropical Diseases. • The molecular mechanism and life cycle of DENV infection have been elucidated. • The inhibitory effect of several heterocyclic Scaffolds on DENV has been outlined. • SAR of heterocycles on the DENV serotypes has been summarized.

Dengue Vaccines: An Update

Abstract: Dengue is one of the most prevalent mosquito-borne diseases in the world, affecting an estimated 390 million people each year, according to models. For the last two decades, efforts to develop safe and effective vaccines to prevent dengue virus (DENV) infections have faced several challenges, mostly related to the complexity of conducting long-term studies to evaluate vaccine efficacy and safety to rule out the risk of vaccine-induced DHS/DSS, particularly in children. At least seven DENV vaccines have undergone different phases of clinical trials; however, only three of them (Dengvaxia®, TV003, and TAK-003) have showed promising results, and are addressed in detail in this review in terms of their molecular design, efficacy, and immunogenicity. Safety-related challenges during DENV vaccine development are also discussed.

Current status and perspectives on vaccine development against dengue virus infection

Abstract: Dengue virus (DENV) consists of four serotypes in the family Flaviviridae and is a causative agent of dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. DENV is transmitted by mosquitoes, Aedes aegypti and A. albopictus, and is mainly observed in areas where vector mosquitoes live. The number of dengue cases reported by the World Health Organization increased more than 8-fold over the last two decades from 505,430 in 2000 to over 2.4 million in 2010 to 5.2 million in 2019. Although vaccine is the most effective method against DENV, only one commercialized vaccine exists, and it cannot be administered to children under 9 years of age. Currently, many researchers are working to resolve the various problems hindering the development of effective dengue vaccines; understanding of the viral antigen configuration would provide insight into the development of effective vaccines against DENV infection. In this review, the current status and perspectives on effective vaccine development for DENV are examined. In addition, a plausible direction for effective vaccine development against DENV is suggested.

Antiviral Agents against Flavivirus Protease: Prospect and Future Direction

Abstract: Flaviviruses cause a significant amount of mortality and morbidity, especially in regions where they are endemic. A recent example is the outbreak of Zika virus throughout the world. Development of antiviral drugs against different viral targets is as important as the development of vaccines. During viral replication, a single polyprotein precursor (PP) is produced and further cleaved into individual proteins by a viral NS2B-NS3 protease complex together with host proteases. Flavivirus protease is one of the most attractive targets for development of therapeutic antivirals because it is essential for viral PP processing, leading to active viral proteins. In this review, we have summarized recent development in drug discovery targeting the NS2B-NS3 protease of flaviviruses, especially Zika, dengue, and West Nile viruses.

Overview of dengue outbreaks in the southwestern Indian Ocean and analysis of factors involved in the shift toward endemicity in Reunion Island: A systematic review

Abstract: Background Dengue is the world’s most prevalent mosquito-borne viral disease. It is endemic in many tropical and subtropical countries and represents a significant global health burden. The first reports of dengue virus (DENV) circulation in the South West Indian Ocean (SWIO) islands date back to the early 1940s; however, an increase in DENV circulation has been reported in the SWIO in recent years. The aim of this review is to trace the history of DENV in the SWIO islands using available records from the Comoros, Madagascar, Mauritius, Mayotte, Seychelles, and Reunion. We focus in particular on the most extensive data from Reunion Island, highlighting factors that may explain the observed increasing incidence, and the potential shift from one-off outbreaks to endemic dengue transmission. Methods Following the PRISMA guidelines, the literature review focused queried different databases using the keywords “dengue” or “Aedes albopictus” combined with each of the following SWIO islands the Comoros, Madagascar, Mauritius, Mayotte, Seychelles, and Reunion. We also compiled case report data for dengue in Mayotte and Reunion in collaboration with the regional public health agencies in these French territories. References and data were discarded when original sources were not identified. We examined reports of climatic, anthropogenic, and mosquito-related factors that may influence the maintenance of dengue transmission independently of case importation linked to travel. Findings and conclusions The first report of dengue circulation in the SWIO was documented in 1943 in the Comoros. Then not until an outbreak in 1976 to 1977 that affected approximately 80% of the population of the Seychelles. DENV was also reported in 1977 to 1978 in Reunion with an estimate of nearly 30% of the population infected. In the following 40-year period, DENV circulation was qualified as interepidemic with sporadic cases. However, in recent years, the region has experienced uninterrupted DENV transmission at elevated incidence. Since 2017, Reunion witnessed the cocirculation of 3 serotypes (DENV-1, DENV-2 and DENV-3) and an increased number of cases with severe forms and deaths. Reinforced molecular and serological identification of DENV serotypes and genotypes circulating in the SWIO as well as vector control strategies is necessary to protect exposed human populations and limit the spread of dengue.

Citywide Integrated Aedes aegypti Mosquito Surveillance as Early Warning System for Arbovirus Transmission, Brazil

Abstract: Infestation indices based on adult trapping predicted dengue outbreaks better than larval indices did.

Dengue and COVID-19: two sides of the same coin

Abstract: Many countries in Asia and Latin America are currently facing a double burden of outbreaks due to dengue and COVID-19. Here we discuss the similarities and differences between the two infections so that lessons learnt so far from studying both infections will be helpful in further understanding their immunopathogenesis and to develop therapeutic interventions.Although the entry routes of the SARS-CoV-2 and the dengue virus (DENV) are different, both infections result in a systemic infection, with some similar clinical presentations such as fever, headache, myalgia and gastrointestinal symptoms. However, while dengue is usually associated with a tendency to bleed, development of micro and macrothrombi is a hallmark of severe COVID-19. Apart from the initial similarities in the clinical presentation, there are further similarities between such as risk factors for development of severe illness, cytokine storms, endothelial dysfunction and multi-organ failure. Both infections are characterised by a delayed and impaired type I IFN response and a proinflammatory immune response. Furthermore, while high levels of potent neutralising antibodies are associated with protection, poorly neutralising and cross-reactive antibodies have been proposed to lead to immunopathology by different mechanisms, associated with an exaggerated plasmablast response. The virus specific T cell responses are also shown to be delayed in those who develop severe illness, while varying degrees of endothelial dysfunction leads to increased vascular permeability and coagulation abnormalities.While there are many similarities between dengue and SARS-CoV-2 infection, there are also key differences especially in long-term disease sequelae. Therefore, it would be important to study the parallels between the immunopathogenesis of both infections for development of more effective vaccines and therapeutic interventions.

The simultaneous crises of dengue and COVID-19 in Pakistan: a double hazard for the country’s debilitated healthcare system

Abstract: Dengue fever is an arthropod-borne viral illness caused by four dengue virus serotypes (DENV 1-4), spread by Aedes mosquitos. Common symptoms of dengue include high peak temperature, headache, myalgia, and malaise following infection, with a rash emerging after 3 to 4 days. More than half of the world's population lives in dengue-endemic areas. Every year, around 100 million dengue cases are reported, with Southeast Asia comprising the majority. Nearly every day, there is a breakout of dengue infections in many areas of Pakistan, in addition to the ongoing COVID-19 pandemic. As a result, combating the dual burden of dengue and COVID-19 is difficult for the nation's healthcare system. Misdiagnosis owing to overlapping symptoms with COVID-19, overburdening of the healthcare system, and a lack of appropriate vaccination are some of the obstacles for dengue infection management. The government of Pakistan is pursuing a variety of measures to combat dengue fever outbreaks, including, The Pakistan Red Crescent Society was asked by the Department of Malaria Control Program to aid in promoting awareness and organizing clean-up campaigns in polluted regions and stagnant water for vector control.

Glycosylation of viral proteins: Implication in virus–host interaction and virulence

Abstract: ABSTRACT Glycans are among the most important cell molecular components. However, given their structural diversity, their functions have not been fully explored. Glycosylation is a vital post-translational modification for various proteins. Many bacteria and viruses rely on N-linked and O-linked glycosylation to perform critical biological functions. The diverse functions of glycosylation on viral proteins during viral infections, including Dengue, Zika, influenza, and human immunodeficiency viruses as well as coronaviruses have been reported. N-linked glycosylation is the most common form of protein modification, and it modulates folding, transportation and receptor binding. Compared to N-linked glycosylation, the functions of O-linked viral protein glycosylation have not been comprehensively evaluated. In this review, we summarize findings on viral protein glycosylation, with particular attention to studies on N-linked glycosylation in viral life cycles. This review informs the development of virus-specific vaccines or inhibitors.

The Aedes aegypti siRNA pathway mediates broad-spectrum defense against human pathogenic viruses and modulates antibacterial and antifungal defenses

Abstract: The mosquito’s innate immune system defends against a variety of pathogens, and the conserved siRNA pathway plays a central role in the control of viral infections. Here, we show that transgenic overexpression of Dicer2 (Dcr2) or R2d2 resulted in an accumulation of 21-nucleotide viral sequences that was accompanied by a significant suppression of dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) replication, thus indicating the broad-spectrum antiviral response mediated by the siRNA pathway that can be applied for the development of novel arbovirus control strategies. Interestingly, overexpression of Dcr2 or R2d2 regulated the mRNA abundance of a variety of antimicrobial immune genes, pointing to additional functions of DCR2 and R2D2 as well as cross-talk between the siRNA pathway and other immune pathways. Accordingly, transgenic overexpression of Dcr2 or R2d2 resulted in a lesser proliferation of the midgut microbiota and increased resistance to bacterial and fungal infections.

Dengue and COVID-19: two sides of the same coin

Abstract: Many countries in Asia and Latin America are currently facing a double burden of outbreaks due to dengue and COVID-19. Here we discuss the similarities and differences between the two infections so that lessons learnt so far from studying both infections will be helpful in further understanding their immunopathogenesis and to develop therapeutic interventions.Although the entry routes of the SARS-CoV-2 and the dengue virus (DENV) are different, both infections result in a systemic infection, with some similar clinical presentations such as fever, headache, myalgia and gastrointestinal symptoms. However, while dengue is usually associated with a tendency to bleed, development of micro and macrothrombi is a hallmark of severe COVID-19. Apart from the initial similarities in the clinical presentation, there are further similarities between such as risk factors for development of severe illness, cytokine storms, endothelial dysfunction and multi-organ failure. Both infections are characterised by a delayed and impaired type I IFN response and a proinflammatory immune response. Furthermore, while high levels of potent neutralising antibodies are associated with protection, poorly neutralising and cross-reactive antibodies have been proposed to lead to immunopathology by different mechanisms, associated with an exaggerated plasmablast response. The virus specific T cell responses are also shown to be delayed in those who develop severe illness, while varying degrees of endothelial dysfunction leads to increased vascular permeability and coagulation abnormalities.While there are many similarities between dengue and SARS-CoV-2 infection, there are also key differences especially in long-term disease sequelae. Therefore, it would be important to study the parallels between the immunopathogenesis of both infections for development of more effective vaccines and therapeutic interventions.

Infection of Endothelial Cells by Dengue Virus Induces ROS Production by Different Sources Affecting Virus Replication, Cellular Activation, Death and Vascular Permeability

Abstract: Exacerbated inflammatory response and altered vascular function are hallmarks of dengue disease. Reactive oxygen species (ROS) production has been associated to endothelial barrier disturbance and microvascular alteration in distinct pathological conditions. Increased ROS has been reported in in vitro models of dengue virus (DENV) infection, but its impact for endothelial cell physiology had not been fully investigated. Our group had previously demonstrated that infection of human brain microvascular endothelial cells (HBMEC) with DENV results in the activation of RNA sensors and production of proinflammatory cytokines, which culminate in cell death and endothelial permeability. Here, we evaluated the role of mitochondrial function and NADPH oxidase (NOX) activation for ROS generation in HBMEC infected by DENV and investigated whether altered cellular physiology could be a consequence of virus-induced oxidative stress. DENV-infected HBMECs showed a decrease in the maximal respiratory capacity and altered membrane potential, indicating functional mitochondrial alteration, what might be related to mtROS production. Indeed, mtROS was detected at later time points after infection. Specific inhibition of mtROS diminished virus replication, cell death, and endothelial permeability, but did not affect cytokine production. On the other hand, inhibition of NOX-associated ROS production decreased virus replication and cell death, as well as the secretion of inflammatory cytokines, including IL-6, IL-8, and CCL5. These results demonstrated that DENV replication in endothelial cells induces ROS production by different pathways, which impacts biological functions that might be relevant for dengue pathogenesis. Those data also indicate oxidative stress events as relevant therapeutical targets to avoid vascular permeability, inflammation, and neuroinvasion during DENV infection.

Label-free electrochemical immunosensor based on l-cysteine-functionalized AuNP on reduced graphene oxide for the detection of dengue virus E-protein in dengue blood serum

Abstract: Dengue fever is one of the most deadly viruses, and it has become a recurring public health problem in tropical areas. The increase in dengue cases worldwide has increased the demand for rapid and accurate diagnostic methods. In order to detect the content of dengue virus type E-proteins (DENV-E protein), we fabricated an efficient label-free electrochemical immunosensor platform that was constructed based on in situ reduction and functionalized gold nanoparticles decorated heteroatom-doped reduced graphene oxides nanocomposites (AuNPs/NSG). Here, L-cystein (L-cys) is introduced to green reducing and stabilizing agent of Au(III) and graphene oxide . The sensing platform offers a suitable higher number of antibody immobilization by providing L-cys. The highly crystalline AuNP was unfoirmly grafting on 2D graphene sheet . Under the optimized conditions, the as-designed immunosensor exhibited a wide linear working range from 0.01–100 ng mL −1 with a low detection limit of 1.6 pg mL −1 for DENV-E detection. The proposed immunosensor shows high selective for discriminating DENV-E against antibodies of their antibodies, including the closely related DENV. The reliability of fabricated immunosensor had been used to detect the viral E-protein contents in serum samples collected from patients, and obtained results were compared with the enzyme-linked immunosorbend assay. Furthermore, the developed AuNPs/NSG immunosensor can be used as a potential probe for future prospective clinical diagnostic applications. The electrochemical detection of dengue virus type E-proteins (DENV–E protein), the efficient label-free electrochemical immunosensor platform was constructed based on in situ reduction and functionalized gold nanoparticles decorated heteroatom doped reduced graphene oxides nanocomposites. The fabricated electrode exhibits lower detection limit and higher sensing and selective detection of DENV-E protein in dengue blood serum. • Label-free, highly selective, and an ultrasensitive label-free electrochemical immunosensor platform for DENV–E protein detection. • In situ reduction and functionalized AuNPs/NSG was synthesized via l -cysteine to achieve excellent electrochemical properties. • l -Cysteine on the nanocomposites to bind covalently with anti-DENV–E protein immobilization. • Immunosensor had a low detection limit of 1.6 pg mL −1 for DENV-E detection. • Successful detection of DENV–E protein in human blood serum samples.

Knowledge gaps in the epidemiology of severe dengue impede vaccine evaluation

Abstract: The most severe consequences of dengue virus infection include shock, haemorrhage, and major organ failure; however, the frequency of these manifestations varies, and the relative contribution of pre-existing anti-dengue virus antibodies, virus characteristics, and host factors (including age and comorbidities) are not well understood. Reliable characterisation of the epidemiology of severe dengue first depends on the use of consistent definitions of disease severity. As vaccine trials have shown, severe dengue is a crucial interventional endpoint, yet the infrequency of its occurrence necessitates the inclusion of thousands of study participants to appropriately compare its frequency among participants who have and have not been vaccinated. Hospital admission is frequently used as a proxy for severe dengue; however, lack of specificity and variability in clinical practices limit the reliability of this approach. Although previous infection with a dengue virus is the best characterised risk factor for developing severe dengue, the influence of the timing between dengue virus infections and the sequence of dengue virus infections on disease severity is only beginning to be elucidated. To improve our understanding of the diverse factors that shape the clinical spectrum of disease resulting from dengue virus infection, prospective, community-based and clinic-based immunological, virological, genetic, and clinical studies across a range of ages and geographical regions are needed.

Risk and predictive factors for severe dengue infection: A systematic review and meta-analysis

Abstract: Background Dengue is a major public health issue worldwide and severe dengue (SD) is life threatening. It is critical to triage patients with dengue infection in the early stage. However, there is limited knowledge on early indicators of SD. The objective of this study is to identify risk factors for the prognosis of SD and try to find out some potential predictive factors for SD from dengue fever (DF) in the early of infection. Methods The PubMed, Cochrane Library and Web of Science databases were searched for relevant studies from June 1999 to December 2020. The pooled odds ratio (OR) or standardized mean difference (SMD) with 95% confidence intervals (CI) of identified factors was calculated using a fixed or random effect model in the meta-analysis. Tests for heterogeneity, publication bias, subgroup analyses, meta-regression, and a sensitivity analysis were further performed. Findings A total of 6,848 candidate articles were retrieved, 87 studies with 35,184 DF and 8,173 SD cases met the eligibility criteria. A total of 64 factors were identified, including population and virus characteristics, clinical symptoms and signs, laboratory biomarkers, cytokines, and chemokines; of these factors, 34 were found to be significantly different between DF and SD, while the other 30 factors were not significantly different between the two groups after pooling the data from the relevant studies. Additionally, 9 factors were positive associated with SD within 7 days after illness when the timing subgroup analysis were performed. Conclusions Practical factors and biomarkers for the identification of SD were established, which will be helpful for a prompt diagnosis and early effective treatment for those at greatest risk. These outcomes also enhance our knowledge of the clinical manifestations and pathogenesis of SD.

Designer DNA nanostructures for viral inhibition

Abstract: Emerging viral diseases can substantially threaten national and global public health. Central to our ability to successfully tackle these diseases is the need to quickly detect the causative virus and neutralize it efficiently. Here we present the rational design of DNA nanostructures to inhibit dengue virus infection. The designer DNA nanostructure (DDN) can bind to complementary epitopes on antigens dispersed across the surface of a viral particle. Since these antigens are arranged in a defined geometric pattern that is unique to each virus, the structure of the DDN is designed to mirror the spatial arrangement of antigens on the viral particle, providing very high viral binding avidity. We describe how available structural data can be used to identify unique spatial patterns of antigens on the surface of a viral particle. We then present a procedure for synthesizing DDNs using a combination of in silico design principles, self-assembly, and characterization using gel electrophoresis, atomic force microscopy and surface plasmon resonance spectroscopy. Finally, we evaluate the efficacy of a DDN in inhibiting dengue virus infection via plaque-forming assays. We expect this protocol to take 2–3 d to complete virus antigen pattern identification from existing cryogenic electron microscopy data, ~2 weeks for DDN design, synthesis, and virus binding characterization, and ~2 weeks for DDN cytotoxicity and antiviral efficacy assays. Star-shaped DNA nanostructures are designed to inhibit dengue virus in vitro. The ‘DNA star’ acts as a template to display ten virus-binding aptamers with precise spatial patterning to mirror the virus antigens, achieving high viral binding avidity.

Reduced dengue incidence during the COVID-19 movement restrictions in Sri Lanka from March 2020 to April 2021

Abstract: Dengue is the major mosquito-borne disease in Sri Lanka. After its first detection in January 2020, COVID-19 has become the major health issue in Sri Lanka. The impact of public health measures, notably restrictions on movement of people to curb COVID-19 transmission, on the incidence of dengue during the period March 2020 to April 2021 was investigated.The incidence of dengue and COVID-19, rainfall and the public movement restrictions implemented to contain COVID-19 transmission were obtained from Sri Lanka government sources. A Seasonal Autoregressive Integrated Moving Average (SARIMA) model was used to predict the monthly dengue incidence from March 2020 to April 2021 for each of the country's 25 districts based on five years of pre-pandemic data, and compared with the actual recorded incidence of dengue during this period. Ovitrap collections of Aedes larvae were performed in Jaffna city in the Jaffna district from August 2020 to April 2021 and the findings compared with similar collections made in the pre-pandemic period from March 2019 to December 2019.The recorded numbers of dengue cases for every month from March 2020 to April 2021 in the whole country and for all 25 districts over the same period were lower than the numbers of dengue cases predicted from data for the five years (2015-2019) immediately preceding the COVID-19 pandemic. The number of dengue cases recorded nationwide represented a 74% reduction from the predicted number of dengue cases for the March 2020 to April 2021 period. The numbers of Aedes larvae collected from ovitraps per month were reduced by 88.6% with a lower proportion of Ae. aegypti than Ae. albopictus in Jaffna city from August 2020 until April 2021 compared with March 2019 to December 2019.Public health measures that restricted movement of people, closed schools, universities and offices to contain COVID-19 transmission unexpectedly led to a significant reduction in the reported numbers of dengue cases in Sri Lanka. This contrasts with findings reported from Singapore. The differences between the two tropical islands have significant implications for the epidemiology of dengue. Reduced access to blood meals and lower vector densities, particularly of Ae. aegypti, resulting from the restrictions on movement of people, are suggested to have contributed to the lower dengue incidence in Sri Lanka.

Designer DNA nanostructures for viral inhibition

Abstract: Emerging viral diseases can substantially threaten national and global public health. Central to our ability to successfully tackle these diseases is the need to quickly detect the causative virus and neutralize it efficiently. Here we present the rational design of DNA nanostructures to inhibit dengue virus infection. The designer DNA nanostructure (DDN) can bind to complementary epitopes on antigens dispersed across the surface of a viral particle. Since these antigens are arranged in a defined geometric pattern that is unique to each virus, the structure of the DDN is designed to mirror the spatial arrangement of antigens on the viral particle, providing very high viral binding avidity. We describe how available structural data can be used to identify unique spatial patterns of antigens on the surface of a viral particle. We then present a procedure for synthesizing DDNs using a combination of in silico design principles, self-assembly, and characterization using gel electrophoresis, atomic force microscopy and surface plasmon resonance spectroscopy. Finally, we evaluate the efficacy of a DDN in inhibiting dengue virus infection via plaque-forming assays. We expect this protocol to take 2–3 d to complete virus antigen pattern identification from existing cryogenic electron microscopy data, ~2 weeks for DDN design, synthesis, and virus binding characterization, and ~2 weeks for DDN cytotoxicity and antiviral efficacy assays. Star-shaped DNA nanostructures are designed to inhibit dengue virus in vitro. The ‘DNA star’ acts as a template to display ten virus-binding aptamers with precise spatial patterning to mirror the virus antigens, achieving high viral binding avidity.

Effects of Acute Dengue Infection on Sperm and Virus Clearance in Body Fluids of Men

Abstract: We investigated the effects of dengue virus (DENV) on semen using samples collected 7, 15, 30, 60, and 90 days after symptom onset from 10 infected volunteers on Réunion Island. We assessed characteristics of semen and reproductive hormones and isolated motile spermatozoa from semen. We assayed semen for DENV using reverse transcription PCR and searched for DENV RNA by virus isolation in Vero E6 cell cultures. Four volunteers had >1 DENV RNA-positive semen samples; 2 volunteers had DENV RNA–positive semen at day 15 and 1 at day 30. No motile sperm were DENV positive. After exposure to positive semen, few Vero E6 cells stained positive for DENV antigens, indicating low levels of replicative virus. We found DENV had shorter duration in semen than in blood. These findings support the possibilities that DENV is sexually transmissible for a short period after acute dengue illness and that acute dengue induces reversible alterations in sperm.

Telmisartan Restricts Chikungunya Virus Infection <i>In Vitro</i> and <i>In Vivo</i> through the AT1/PPAR-γ/MAPKs Pathways

Abstract: Chikungunya virus (CHIKV) has reemerged as a global public health threat. The inflammatory pathways of the renin-angiotensin system (RAS) and peroxisome proliferator-activated receptor-gamma (PPAR-γ) are usually involved in viral infections. Thus, telmisartan (TM), which is known to block the angiotensin 1 (AT1) receptor and activate PPAR-γ, was investigated for activity against CHIKV. The anti-CHIKV effect of TM was investigated in vitro (Vero cells, RAW 264.7 cells, and human peripheral blood mononuclear cells [hPBMCs]) and in vivo (C57BL/6 mice). TM was found to abrogate CHIKV infection efficiently (50% inhibitory concentration (IC50) of 15.34 to 20.89 μM in the Vero cells and RAW 264.7 cells, respectively). Viral RNA and proteins were reduced remarkably. Additionally, TM interfered in the early and late stages of the CHIKV life cycle with efficacy during pretreatment and posttreatment. Moreover, the agonist of the AT1 receptor and an antagonist of PPAR-γ increased CHIKV infection, suggesting that the antiviral potential of TM occurs through modulating host factors. In addition, reduced activation of all major mitogen-activated protein kinases (MAPKs), NF-κB (p65), and cytokines by TM occurred through the inflammatory axis and supported the fact that the anti-CHIKV efficacy of TM is partly mediated through the AT1/PPAR-γ/MAPKs pathways. Interestingly, at a human equivalent dose, TM abrogated CHIKV infection and inflammation significantly, leading to reduced clinical scores and complete survival of C57BL/6 mice. Additionally, TM reduced infection in hPBMC-derived monocyte-macrophage populations in vitro. Hence, TM was found to reduce CHIKV infection by targeting both viral and host factors. Considering its safety and in vivo efficacy, it can be a suitable candidate in the future for repurposing against CHIKV.

Envelope E protein of dengue virus and phospholipid binding to the late endosomal membrane.

Abstract: Flaviviruses include many significant human pathogens, comprising dengue, West Nile, Yellow fever, Japanese encephalitis, Zika and tick-borne encephalitis viruses and many others, affecting millions of people in the world. These viruses have produced important epidemics in the past, they continue to do it and they will undoubtedly continue to do so in the future. Flaviviruses enter into the cells via receptor-mediated endocytosis by fusing its membrane with the endosomal membrane in a pH-dependent manner with the help of the envelope E protein, a prototypical class II membrane fusion protein. The envelope E protein has a conserved fusion peptide at its distal end, which is responsible in the first instance of inserting the protein into the host membrane. Since the participation of other segments of the E protein in the fusion process should not be ruled out, we have used atomistic molecular dynamics to study the binding of the distal end of domain II of the envelope E protein from Dengue virus (DENV) with a complex membrane similar to the late-endosome one. Our work shows that not only the fusion peptide participates directly in the fusion, but also two other sequences of the protein, next to the fusion peptide it in the three-dimensional structure, are jointly wrapped in the fusion process. Overall, these three sequences represent a new target that would make it possible to obtain effective antivirals against DENV in particular and Flaviviruses in general.

Dengue Virus NS1 Uses Scavenger Receptor B1 as a Cell Receptor in Cultured Cells

Abstract: Dengue is the most common viral disease transmitted to humans by mosquitoes. The dengue virus NS1 is a multifunctional glycoprotein necessary for viral replication. ABSTRACT The dengue virus NS1 is a multifunctional protein that forms part of replication complexes. NS1 is also secreted, as a hexamer, to the extracellular milieu. Circulating NS1 has been associated with dengue pathogenesis by several mechanisms. Cell binding and internalization of soluble NS1 result in endothelial hyperpermeability and in the downregulation of the innate immune response. In this work, we report that the HDL scavenger receptor B1 (SRB1) in human hepatic cells and a scavenger receptor B1-like in mosquito C6/36 cells act as cell surface binding receptors for dengue virus NS1. The presence of the SRB1 on the plasma membrane of C6/36 cells, as well as in Huh7 cells, was demonstrated by confocal microscopy. The internalization of NS1 can be efficiently blocked by anti-SRB1 antibodies, and previous incubation of the cells with HDL significantly reduces NS1 internalization. Significant reduction in NS1 internalization was observed in C6/36 cells transfected with siRNAs specific for SRB1. In addition, the transient expression of SRB1 in Vero cells, which lacks the receptor, allows NS1 internalization in these cells. Direct interaction between soluble NS1 and the SRB1 in Huh7 and C6/36 cells was demonstrated in situ by proximity ligation assays and in vitro by surface plasmon resonance. Finally, results are presented indicating that the SRB1 also acts as a cell receptor for Zika virus NS1. These results demonstrate that dengue virus NS1, a bona fide lipoprotein, usurps the HDL receptor for cell entry and offers explanations for the altered serum lipoprotein homeostasis observed in dengue patients. IMPORTANCE Dengue is the most common viral disease transmitted to humans by mosquitoes. The dengue virus NS1 is a multifunctional glycoprotein necessary for viral replication. NS1 is also secreted as a hexameric lipoprotein and circulates in high concentrations in the sera of patients. Circulating NS1 has been associated with dengue pathogenesis by several mechanisms, including favoring of virus replication in hepatocytes and dendritic cells and disruption of the endothelial glycocalyx leading to hyperpermeability. Those last actions require NS1 internalization. Here, we identify the scavenger cell receptor B1, as the cell-binding receptor for dengue and Zika virus NS1, in cultured liver and in mosquito cells. The results indicate that flavivirus NS1, a bona fide lipoprotein, usurps the human HDL receptor and may offer explanations for the alterations in serum lipoprotein homeostasis observed in dengue patients.