Combining antiviral drugs with different mechanisms of action can help prevent the development of resistance by attacking the infectious agent through multiple pathways. Additionally, by using faster and more economical screening methods, effective synergistic drug candidates can be rapidly identified, facilitating faster paths to clinical testing. In this work, a rapid method was standardized to identify possible synergisms from drug combinations. We analyzed the possible reduction of antiviral effective concentration of drugs already approved by the FDA, such as ivermectin (IVM), ribavirin (RIBA) and acyclovir (ACV) against Zika virus (ZIKV), Chikungunya virus (CHIKV) and herpes virus type 2 (HHV-2). Essential oils (EOs) were also included in the study since they have been reported for more than a couple of decades to have broad-spectrum antiviral activity. We also continued studying the antiviral properties of one of our patented molecule with broad-spectrum antiviral activity, the ferruginol analogue 18-(phthalimid-2-yl)ferruginol (phthFGL). In general, the combination of IVM, phthFGL and Oregano EO showed the greatest synergism potential against CHIKV, ZIKV and HHV-2, obtaining a reduction in the EC99 value of up to ~8, and ~27, and ~12-fold for CHIKV, in example, respectively. The ternary combination RIBA, phthFGL, Oregano EO was slightly more efficient than the binary combination RIBA/phthFGL but much less efficient than IVM, phthFGL, Oregano EO which indicates that IVM could contribute more to the differentiations of cell targets (for example by inhibition of host heterodimeric importin IMP α/β1 complex) than ribavirin. PhthFGL showed a good pharmacokinetic profile.

Background The COVID-19 pandemic has focused research on the detrimental impact of neuroinvasive single-strand RNA (ssRNA) viruses. Those viruses cause a wide range of human diseases ranging from mild to severe life-threatening conditions, and host factors such as age, sex, and the expression of virus receptors can influence the severity of the infection. For example, older adults and males have a bias for severe disease with COVID-19 infection, but females have a higher risk of Long-COVID. However, there is a gap in understanding the expression of receptors for ssRNA viruses in the human brain and how that expression varies by age and sex. Here, we studied the expression of ssRNA virus receptors in the human brain and compared lifespan changes in females and males to identify age- and sex-specific book patterns. Methods We used a publicly available transcriptomic database of human brain development to characterize the development of 67 viral host factor genes for 10 ssRNA virus families. A data-driven approach was applied to characterize the lifespan trajectories for those ssRNA receptors using samples from 15 brain areas (n=700, F=306, M=394, age range:4 mo-82 yrs). Then, high-dimensional trajectory analyses and visualizations, including tSNE, were used to compare lifespan changes for females and males among the 15 brain areas. Results Unsupervised hierarchical clustering of the 2010 trajectories identified 25 different patterns with a range of sex bias from ones with only female data to ones with only male data. Differential Expression Sliding Window Analysis (DE-SWAN) found a brain-wide pattern of virus receptor sex differences in childhood, before puberty. The virus families had a range of sexual dimorphism, with the Pneumoveridae family being the most dimorphic. Finally, high dimensional visualization of ssRNA virus receptor development in the 15 brain areas showed that the cortex is distinct for females and males. Conclusions Females are often described as developing precociously relative to males, but our findings paint a different picture. Instead, we found that sex differences in virus receptor development in the human brain do not reflect simply linear shifts and are best described as females and males following distinct high-dimensional trajectories. Highlights We studied 15 areas of the human brain to examine sex differences in the development of ssRNA virus receptors. Application of high-dimensional data analyses identified 25 distinct patterns of developmental trajectories for ssRNA virus receptors. There was little overlap in the trajectories between females and males. Most sex differences were in childhood, before puberty. There was variable sexual dimorphism across virus families, with Pneumoveridae being highly dimorphic. Differential Expression Sliding Window Analysis (DE-SWAN) identified brain-wide virus receptor sex differences in childhood. Density-preserving tSNE visualization showed distinct virus receptor development in the cortex for females and males. Contrary to the notion that females develop preciously compared with males, our findings show distinct sex-specific development of virus receptors in the human brain. Plain english summary The COVID-19 pandemic has drawn attention to viruses that can affect the brain, causing various brain-related health issues. Age, sex, and the expression of virus receptors can influence the severity of these infections. For example, older adults and males tend to experience more severe COVID-19 symptoms, while women are more prone to long-lasting effects. However, little is known about virus receptors in the human brain, their age-related changes, and potential sex differences. To address this, we studied the development of 67 genes related to brain-infecting viruses and analyzed data from 15 brain regions in individuals of various ages. Our findings revealed that these genes develop differently in females and males, with the most differences observed in childhood before puberty. Some virus families exhibited more pronounced sex distinctions, notably pneumonia-related virus receptors. Contrary to the belief that females mature more quickly, our study suggests that virus receptors have unique developmental trajectories for females and males.

Flaviviridae is a family of positive-stranded RNA viruses, including human pathogens such as Japanese encephalitis virus (JEV), dengue virus (DENV), zika virus (ZIKV), and West Nile virus (WNV). Nuclear localization of the viral core protein is conserved among Flaviviridae , and this feature may be targeted for developing broad-ranging anti-flavivirus drugs. However, the mechanism of core protein translocation to the nucleus and the importance of nuclear localization in the viral life cycle remain unknown. We aimed to identify the molecular mechanism underlying core protein nuclear translocation. We identified importin-7 (IPO7), an importin-β family protein, as a nuclear carrier for Flaviviridae core proteins. Nuclear import assays revealed that core protein was transported into the nucleus via IPO7, whereas IPO7 deletion by CRISPR/Cas9 impaired their nuclear localization. To understand the importance of core protein nuclear localization, we evaluated the production of infectious virus or single-round-infectious-particles in wild-type or IPO7-deficient cells; both processes were significantly impaired in IPO7-deficient cells, whereas intracellular infectious virus levels were equivalent in wild-type and IPO7-deficient cells. These data suggested that IPO7-mediated nuclear localization of core proteins plays a role in the release of infectious virus particles of flaviviruses.

Background: Zika virus (ZIKV) infection in pregnancy can cause brain and eye abnormalities and neurodevelopmental sequelae. In the absence of medical countermeasures, behavioral interventions were recommended to prevent mosquito bites and sexual transmission of ZIKV. Methods: We used self-reported data from the Zika en Embarazadas y Niños (ZEN) prospective cohort study in Colombia to describe the knowledge, attitudes, and behaviors related to ZIKV prevention in male partners compared to those of their pregnant partners at study enrollment during February 2017 – February 2018. Results: Most male partners reported wearing protective clothing such as long pants (97.6%) and long sleeves (72.8%), as well as covering ankles and feet (89.1%) to prevent ZIKV infection. When comparing the preventive behavior of condom use between male and pregnant partners, 26 pairs (10.0%) both responded that they performed the behavior. Overall, 25.1% of male partners and 18.9% of pregnant people reported any condom use during the three months before enrolling in ZEN. When comparing other preventive behaviors between male and pregnant partners, the behavior which was most frequently reported by both partners was wearing long pants (85.4%), and the least frequently reported by both partners was using condoms after finding out about a partner’s pregnancy (3.4%). Conclusions: These findings provide new information on the behaviors used by male partners to prevent ZIKV infection during the outbreak in Colombia and can be used to inform future ZIKV and other arbovirus prevention strategies that include both pregnant people and their partners.

SARS-CoV-2 can trigger autoimmune central nervous system (CNS) diseases in genetically susceptible individuals, a mechanism poorly understood. Molecular mimicry (MM) has been identified in other viral diseases as potential triggers of autoimmune CNS events. This study investigated if MM is the process through which SARS-CoV-2 induce the breakdown of immune tolerance. The frequency of autoimmune CNS disorders was evaluated in a prospective cohort with patients admitted in the COVID-19 Intense Care Unity (ICU) in Rio de Janeiro. Then, an in silico analysis was performed to identify the conserved regions which share high identity between SARS-Cov-2 anti-gens and human proteins. The sequences with significant identity and antigenic properties were then assessed for their binding capacity to HLA subtypes. Of the 112 patients included, 3 were classified as having an autoimmune disorder. A total of eleven combinations had significant linear and three-dimensional overlap. NMDAR1, MOG and MPO were the self-antigens with more significant combinations, followed by GAD65. All sequences presented at least one epitope with strong or intermediate binding capacity to the HLA subtypes selected. This study underscores the possibility that CNS autoimmune attacks observed in COVID-19 patients, including those in our population, could be driven by MM in genetically predisposed individuals.

Le Dantec virus (LDV), type species of the genus Ledantevirus within the Rhabdoviridae has been associated with human disease but has gone undetected since the 1970s. We describe the detection of LDV in a human case of undifferentiated fever in Uganda by metagenomic sequencing and demonstrate a serological response using ELISA and pseudotype neutralisation. By screening a cohort of 997 individuals sampled in 2016, we show frequent exposure to ledanteviruses with 76% of individuals seropositive in Western Uganda, but lower seroprevalence in other areas. Serological cross-reactivity as measured by pseudotype-based neutralisation was confined to Ledantevirus, indicating population seropositivity may represent either exposure to LDV or related ledanteviruses. We also describe the discovery of a closely related ledantevirus, Odro virus, in the synanthropic rodent Mastomys erythroleucus . Ledantevirus infection is common in Uganda but is geographically heterogenous. Further surveys of patients presenting with acute fever are required to determine the contribution of these emerging viruses to febrile illness in Uganda.

Background: Cross-species horizontal gene transfer (HGT) involves the transfer of genetic material between different species of organisms. In recent years, mounting evidence has emerged that cross-species HGT does take place and may play a role in the development and progression of diseases. Methods: : Transcriptomic data obtained from patients with gallbladder cancer (GBC)was assessed for the differential expression of antisense RNAs (asRNAs). Cross-species blast analysis was performed with viral, bacterial, fungal, and ancient human genomes to elucidate the evolutionary cross species origins of these differential asRNAs. Functional enrichment analysis and text mining were conducted and a network of asRNAs targeting mRNAs was constructed to understand the function of differential asRNAs better. Results: : A total of 17 differentially expressed antisense RNAs (asRNAs) were identified in gallbladder cancer tissue. BLAST analysis of 15 of these asRNAs (AFAP1-AS1, HMGA2-AS1, MNX1-AS1, SLC2A1-AS1, BBOX1-AS1, ELFN1-AS1, TRPM2-AS, DNAH17-AS1, DCST1-AS1, VPS9D1-AS1, MIR1-1HG-AS1, HAND2-AS1, PGM5P4-AS1, PGM5P3-AS1, and MAGI2-AS) showed varying degree of similarities with bacterial and viral genomes, except for UNC5B-AS1 and SOX21-AS1, which were conserved during evolution. Two of these asRNAs (VPS9D1-AS1 and SLC2A1-AS1) exhibited a high degree of similarity with viral genomes including (Chikungunya virus, Human immunodeficiency virus 1, Stealth virus 1, and Zika virus) and bacterial genomes including ( Staphylococcus sp., Bradyrhizobium sp., Pasteurella multocida sp., and , Klebsiella pneumoniae sp .), indicating potential HGT during evolution. Conclusion: The results provide novel evidence supporting the hypothesis that differentially expressed asRNAs in GBC exhibit varying sequence similarity with bacterial, viral, and ancient human genomes, indicating a potential shared evolutionary origin. These non-coding genes are enriched with methylation and were found to be associated with cancer-related pathways, including the P53 and PI3K-AKT signaling pathways, suggesting their possible involvement in tumor development.

Flaviviruses are arthropod-borne (arbo)viruses which can emerge rapidly and cause explosive epidemics of severe disease. Some of the most epidemiologically important flaviviruses, including dengue virus (DENV), Zika virus (ZIKV) and yellow fever virus (YFV), are transmitted by Aedes mosquitoes, most notably Aedes aegypti and Aedes albopictus. After a mosquito blood feeds on an infected host, virus enters the midgut and infects the midgut epithelium. The virus must then overcome a series of barriers before reaching the mosquito saliva and being transmitted to a new host. The virus must escape from the midgut (known as the midgut escape barrier; MEB), which is thought to be mediated by transient changes in the permeability of the midgut-surrounding basal lamina layer (BL) following blood feeding. Here, we present a mathematical model of the within-mosquito population dynamics of flaviviruses that includes the interaction of the midgut and BL which can account for the MEB. Our results indicate a dose-dependency of midgut establishment of infection as well as rate of escape from the midgut: collectively, these suggest that the extrinsic incubation period (EIP) – the time taken for DENV virus to be transmissible after infection – is shortened when mosquitoes imbibe more virus. Additionally, our experimental data indicates that multiple blood feeding events, which more closely mimic mosquito-feeding behavior in the wild, can hasten the course of infections, and our model predicts that this effect is sensitive to the amount of virus imbibed. Our model indicates that mutations to the virus which impact its replication rate in the midgut could lead to even shorter EIPs when double-feeding occurs. Mechanistic models of within-vector viral infection dynamics provide a quantitative understanding of infection dynamics and could be used to evaluate novel interventions that target the mosquito stages of the infection. Author summary Aedes mosquitoes are the main vectors of dengue virus (DENV), Zika virus (ZIKV) and yellow fever virus (YFV), all of which can cause severe disease in humans with dengue alone infecting an estimated 100-400 million people each year. Understanding the processes that affect whether, and at which rate, mosquitoes may transmit such viruses is, hence, paramount. Here, we present a mathematical model of virus dynamics within infected mosquitoes. By combining the model with novel experimental data, we show that the course of infection is sensitive to the initial dose of virus ingested by the mosquito. The data also indicates that mosquitoes which blood feed subsequent to becoming infected may be able to transmit infection earlier, which is reproduced in the model. This is important as many mosquito species feed multiple times during their lifespan and, any reduction in time to dissemination will increase the number of days that a mosquito is infectious and so enhance the risk of transmission. Our study highlights the key and complementary roles played by mathematical models and experimental data for understanding within-mosquito virus dynamics.

Zika virus and dengue virus are co-circulating flaviviruses with a widespread endemic range. Eliciting broad and potent neutralizing antibodies is an attractive goal for developing a vaccine to simultaneously protect against these viruses. However, the capacity of viral mutations to confer escape from broadly neutralizing antibodies remains undescribed, due in part to limited throughput and scope of traditional approaches. Here, we use deep mutational scanning to map how all possible single amino acid mutations in Zika virus envelope protein affect neutralization by antibodies of varying breadth and potency. While all antibodies selected viral escape mutations, the mutations selected by broadly neutralizing antibodies conferred less escape relative to those selected by narrow, virus-specific antibodies. Surprisingly, even for broadly neutralizing antibodies with similar binding footprints, different single mutations led to escape, indicating distinct functional requirements for neutralization not captured by existing structures. Additionally, the antigenic effects of mutations selected by broadly neutralizing antibodies were conserved across divergent, albeit related, flaviviruses. Our approach identifies residues critical for antibody neutralization, thus comprehensively defining the as-yet-unknown functional epitopes of antibodies with clinical potential. Importance The wide endemic range of mosquito-vectored flaviviruses – such as Zika virus and dengue virus serotypes 1-4 – places hundreds of millions of people at risk of infection every year. Despite this, there are no widely available vaccines, and treatment of severe cases is limited to supportive care. An avenue towards development of more widely applicable vaccines and targeted therapies is the characterization of monoclonal antibodies that broadly neutralize all these viruses. Here, we measure how single amino acid mutations in viral envelope protein affect neutralizing antibodies with both broad and narrow specificities. We find that broadly neutralizing antibodies with potential as vaccine prototypes or biological therapeutics are quantifiably more difficult to escape than narrow, virus-specific neutralizing antibodies.

Epidemiological surveillance is a critical tool to monitor the population's health and facilitate the prevention and control of infectious disease outbreaks. Bacterial infections are well known as one of the leading causes of global infection-related morbidity and mortality. Our study summarizes the number of bacterial infectious diseases in Saudi Arabia, along with an overview and recent advances in treatment or prevention modalities against these reported bacterial infections. This study only covers the reported bacterial infectious diseases in the Saudi Monthly Epidemiology Reports between 2018 and 2021. The results revealed that brucellosis, tuberculosis and salmonellosis were the most frequently reported bacterial infectious diseases in Saudi Arabia. Generally, males were more affected by bacterial infections than females. There was a variation in the distribution of bacterial infectious diseases between Saudi and non-Saudi citizens. Brucellosis and Salmonellosis infections were more common among Saudi citizens, while Tuberculosis was more common in non-Saudis. Interestingly, there was a decline in the incidence rates of numerous bacterial infectious diseases during the Coronavirus Disease 2019 (COVID-19) pandemic and COVID-19 restrictions. However, this decline in the incidence rates might be a result of underreporting during the national lockdown. Some bacterial infectious diseases were rarely reported in Saudi Arabia, including Syphilis, Diphtheria, and Guillain-Barré syndrome.

Zika virus (ZIKV) is a mosquito-borne flavivirus that is emerging and re-emerging. Since 2010, sporadic ZIKV cases have been reported annually in Thailand, with over 1,000 confirmed positive cases in 2016. High-throughput sequencing technologies, such as whole genome sequencing (WGS), have recently enabled the rapid sequencing of pathogen genomes. Therefore, this study used multiplex amplicon sequencing by the Illumina Miseq to describe the ZIKV WGS. Six ZIKV WGS were obtained from three samples of field-caught Culex quinquefasciatus mosquitoes and three urine samples collected from human in three different provinces of Thailand. Additionally, one ZIKV isolate was successfully isolated from a female Cx. quinquefasciatus . Based on WGS analysis, the timing of the 2020 outbreak correlates with the acquisition of five amino acid changes in the Asian lineage ZIKV strains from Thailand 2006, Cambodia 2010 and 2019, and Philippines 2012, including C-T106A, prM-V1A, E-V473M, NS1-A188V, and NS5-M872V. This mutation was found in all seven WGS, which was previously reported to be associated with significantly higher mortality rates. Moreover, phylogenetic analysis indicated that the seven ZIKV sequences belonged to the Asian lineage. The genomic region of the E gene showed the highest nucleotide diversity (0.7–1.3%). The data can be applied to develop molecular tools to better understand the virus's patterns and evolution. Furthermore, it could indicate potential targets for development of more effective to control and prevent the zika outbreak.

Arboviruses represents a real public health problem globally and in the Central African subregion in particular that represents a high risk zone for the emergence and re-emergence of arboviruses outbreaks. Furthermore, an updated review on the current arboviruses burden and associated mosquito vectors is lacking for this region. To contribute in filling this knowledge gap, the current study was designed with following objectives: (i) to systematically review data on the occurrence and distribution of arboviruses and mosquito fauna and (ii) to identify potential spillover mosquito species in Central African region in the last 30 years. Web search enabled the documentation of 2454 articles from different online databases. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and The Quality of Reporting of Meta-analyses (QUORUM) steps for a systematic review enabled the selection of 164 articles that fulfilled our selection criteria. Of the six arboviruses (Dengue Virus (DENV), Chikungunya Virus (CHIKV), Yellow Fever Virus (YFV), Zika Virus (ZIKV), Rift Valley Fever Virus (RVFV), and West Nile Virus (WNV)) of public health concern studied, the most frequently reported were Chikungunya and Dengue. The entomological records showed >248 species of mosquitoes and regrouped under 15 genera with Anopheles (n= 100 species), Culex (n= 56 species) and Aedes (n=52 species) having high species diversity. Three genera were rarely represented with only one species and included: Orthopodomyia, Lutzia, and Verrallina, but individuals of the genera Toxorhinchites and Finlayas were not identified upto the species level. We found that two Aedes species (Ae. aegypti and Ae. albopictus) colonised the same microhabitat and were involved in major epidemics of the six medically important arboviruses and other less frequently identified mosquito-genera consisted of competent species and were associated with outbreaks of medical and zoonotic arboviruses. The present study reveals high species richness of competent mosquito-vectors that could lead to the spillover of medically important arboviruses in the region. Although epidemiological studies were found, they were not regularly documented and this also applies to vector competence and transmission studies. Future studies will consider unpublished information in dissertations, technical reports from different countries to enable it more consistent. A regional project entitled: Ecology of Arboviruses (EcoVir) is underway in three countries (Gabon, Benin and Cote d’Ivoire) to generate a more comprehensive epidemiological and entomological data on this topic.

The COVID-19 pandemic caused by SARS-CoV-2 becomes a serious threat to global health and requires the development of effective antiviral therapies. Current therapies that target viral proteins have limited efficacy with side effects. In this study, we investigated the antiviral activity of MIT-001, a small molecule reactive oxygen species (ROS) scavenger targeting mitochondria, against SARS-CoV-2 and other zoonotic viruses in vitro. The antiviral activity of MIT-001 was quantified by RT-qPCR and plaque assay. We also evaluated the functional analysis of MIT-001 by JC-1 staining to measure mitochondrial depolarization, total RNA sequencing to investigate gene expression changes, and immunoblot to quantify protein expression levels. The results showed that MIT-001 effectively inhibited the replication of B.1.617.2 and BA.1 strains, Zika virus, Seoul virus, and Vaccinia virus. Treatment with MIT-001 restored the expression of heme oxygenase-1 (HMOX1) and NAD(P)H: quinone oxidoreductase 1 (NqO1) genes, anti-oxidant enzymes reduced by SARS-CoV-2, to normal levels. The presence of MIT-001 also alleviated mitochondrial depolarization caused by SARS-CoV-2 infection. These findings highlight the potential of MIT-001 as a broad-spectrum antiviral compound that targets for zoonotic RNA and DNA viruses, providing a promising therapeutic approach to combat viral infection.

Coronaviruses contain one of the largest genomes among the RNA viruses, coding for 14-16 non-structural proteins (nsp) involved in proteolytic processing, genome replication and transcription and four structural proteins that build the core of the mature virion. Due to conservation across coronaviruses nsps form a group of promising drug targets as their inhibition directly affects viral replication and therefore progression of infection. A minimal but fully functional replication and transcription complex was shown to be formed by one RNA-dependent RNA polymerase (nsp12), one nsp7, two nsp8 accessory subunits, and two helicase (nsp13) enzymes. Due to its key role in viral replication and lack of close host homologs nsp12 has the potential to be a safe and therapeutically attractive broad spectrum anti-viral target. Nsp13 provides multiple starting points for development of inhibitors as it exhibits two functional properties, the NTPase and unwinding activity, and direct interactions with the RNA-dependent RNA polymerase complex. Our approach involved, targeting both nsp12 together with nsp13 to allow multiple starting point to interfere with virus infection progression. Here we report a combined in-vitro repurposing screening approach, identifying new and confirming reported SARS-CoV-2 nsp13 and nsp12 inhibitors

Despite historical experience and health state capacity, Brazil struggled to address the COVID-19 pandemic. Whereas past administrations have proactively mobilized resources to combat epidemics, Bolsonaro’s administration took a more passive role. The federal government’s relative absence forced state and municipal governments to plan and manage the pandemic response. At the state level, governors, across different political parties and varying loyalties towards Bolsonaro, banded together to procure vaccinations for their citizens. At the municipal level, mayors enacted a series of social and public health support measures. However, these responses were not universal, and there has been significant variation between and within municipalities. What explains municipal variation in the enactment of pandemic-related public policies? Current studies focus on three explanations – political alignment, health state capacity, and diffusion – to explain policy implementation variation across and within countries at various levels of governance. While these studies are important to our knowledge of pandemic responses at the federal and state levels, there have been few studies examining these explanations at the local level. Using quantitative and qualitative evidence, this article argues that, while conventional wisdom holds in explaining pandemic responses, the mayor’s health training and population size mediate conventional explanations such as state capacity and political polarization. This article makes two key contributions. First, it tests the conventional wisdom of pandemic responses at the municipal level, which is important to our understanding of local city dynamics and highlights key mechanisms that mediate the implementation of COVID-19 policy. Second, it provides alternative hypotheses, such as the mayor’s health training and population size, which are idiosyncratic to the level of analysis. This project is part of a larger project exploring the political motivations of COVID-19 implementation in Brazilian municipalities.

ABSTRACT In India, the SARS-CoV-2 Delta wave (2020-21) gradually faded away with the advent of the Omicron variants (2021-present). Dengue incidences were observably less in Southeast Asia during the active years of the pandemic (2020-21). However, Dengue virus type 3 (DV3) cases were increasingly reported in India and many other dengue-endemic countries concurrent with the progression of the Omicron wave since 2022. This observation prompted us to investigate the current state of cross-reactivity between prevalent SARS-CoV-2 variants and different DV serotypes. Fifty-five COVID-19 serum samples (collected between January-September, 2022) and three pre-pandemic healthy serum samples were tested for DV or SARS-CoV-2 Immunoglobulin G/Immunoglobulin M (IgG/IgM) using the lateral flow immunoassays (LFIAs). The SARS-CoV-2 antibody (Ab)-positive samples were further tested for their ability to cross-neutralize DV types 1-4 (DV1-4) in Huh7 cell lines. Cross-reactivity between SARS-CoV-2 and DV diminished with the shift from Delta to Omicron prevalence. COVID-19 serum samples that were DV cross-reactive neutralized all DV serotypes, including DV3. However, Omicron wave serum samples were predominantly DV non-cross-reactive (about 70%) in LFIAs and coincided with the prevalence of BA.2 Omicron variant. They also cross-neutralized DV1, 2 and 4 but enhanced DV3 infectivity as evident from increased DV3 titres in virus neutralization test (VNT) compared to control serum samples. In conclusion, DV non-cross-reactive COVID-19 serum samples are becoming increasingly prominent in the present times. These non-cross-reactive serums could neutralize DV1, 2 and 4 but they are contributing to the surge in DV3 cases worldwide by means of ADE.

Japanese encephalitis infection is a vector-borne disease caused by the flavivirus Japanese encephalitis virus (JEV). It is responsible for severe brain infections in humans worldwide. Given the ubiquitous nature of complications and tropism associated with Japanese encephalitis (JE) infection, a holistic understanding of its molecular mechanism is essential. The phenomenon of abnormal protein aggregation into pathogenic amyloids is now increasingly linked to multiple human diseases, also known as Amyloidosis. Most are neurodegenerative disorders, but amyloidosis is not restricted to a specific organ or tissue type. The overlap of viral protein aggregation with human pathologies remains limited, and it is gaining momentum, especially after the devastating Covid-19 pandemic. Therefore, in this study, we have examined the likelihood of aggregation for the entire collection of proteins in JEV. Multiple independent web server tools were employed to scan for potential amyloid-prone regions (APRs), and it was followed by in vitro validation using two JEV transmembrane domains, Capsid anchor, and 2K peptides. These synthetic viral peptides were introduced to artificial aggregation-inducing conditions and then analyzed using different dye-based assays and microscopy methods confirming amyloid-like fibril structure formation. We found these aggregates cytotoxic to human neuronal cell lines and membrane damaging to human blood-derived RBCs. The aggregation kinetics of both peptides is enhanced in the presence of artificial membrane models and seeds of self and diabetes hallmark protein Amylin. Our findings thereby strongly suggest the possibility of JEV protein aggregation playing a vital role in its pathogenesis, opening up a broad scope of future study. Also, the interplay between JEV protein aggregation and initiation/progression of other proteopathies is possible and needs further exploration.

The prevalence of mosquito-borne diseases has precipitated significant global health concerns, particularly in tropical regions where millions of individuals remain at a heightened risk of infection. These diseases, often transmitted through the bites of infected mosquitoes, can lead to serious illnesses, such as malaria, dengue fever, Zika virus, and chikungunya. Given the substantial health and economic burden imposed by these diseases, there is an undeniable need to effectively address this issue. The efficiency of the selected weed plant extracts in water was evaluated for larval mortality of Culex quinquefasciatus ( Cu. quinquefasciatus ) after 24 h. The LC 50 and LC 95 values of the plant extracts of Carthamus oxycantha , Trachyspermum ammi , Achyranthes aspera , Convolvulus arvensis , and Cynodon dactylon were also determined. The experiment was conducted in a completely randomized design with six treatments, each replicated three times. The plant extracts were tested at five concentrations: 200, 300, 500, 700, and 1000 ppm. Carthamus oxycantha in the water extract yielded a higher mortality of 42% after 24 h. Both LC 50 and LC 95 values were maximum for Cynodon dactylon (66749.45) and lowest for Carthamus oxycantha (2535.34), whereas the LC 95 value was highest for Cynodon dactylon . The study recommends that these weed plant extracts be tested at lower concentrations, as higher concentrations yield 100% mortality of the pest. Natural bio-products are not only favored in vector control measures but also in resistance developed by the larvae. These findings highlight the potential of natural bioproducts for vector control and addressing larval resistance, emphasizing the need to identify new active compounds from cost-effective natural sources against mosquitoes.

Despite the exponential rise of social media and online news sources, traditional news media, such as newspapers, remain essential for health information. During a health crisis, such as a pandemic, the news media can play a significant role by conveying messages from emergency management agencies to the general population. In this study, we conducted a qualitative content analysis of the New York Times (NYT) coverage of COVID-19-related stories published between 1, 2020, and August 31, 2022. Based on crisis communication and media framing theories, we analyzed the coverage span, size, and wording of the articles. Based on our sample, we observed that the frequency of COVID-19-related reports was higher at the beginning of the pandemic and decreased over time. Section analysis of the articles reflected a high diversity as coronavirus-related stories were distributed in more than ten sections. In addition, we found that almost 16 percent of the articles were placed in U.S. news and 14 percent were in the editorial. It also indicated a national interest and the topic's significance as articles found in editorials received more attention. Regarding framing, we discovered that the NYT coverage of COVID-19 dealt more with human interest, responsibility, and conflict than economic and morality frames.

Background: The mosquito microbiome is an important modulator of vector competence and vectoral capacity. Unlike the extensively studied bacterial microbiome, fungal communities in the mosquito microbiome (mycobiome) remain largely unexplored. To work towards getting an improved understanding of the fungi associated with mosquitoes, we sequenced the mycobiome of three field-collected and laboratory-reared mosquito species ( Aedes albopictus, Aedes aegypti , and Culex quinquefasciatus ). Results Our analysis showed both environment and host species were contributing to the diversity of the fungal microbiome of mosquitoes. When comparing species, Ae. albopictus possessed a higher number of diverse fungal taxa than Cx. quinquefasciatus , while strikingly less than 1% of reads from Ae. aegypti samples were fungal. Fungal reads from Ae. aegypti were < 1% even after inhibiting host amplification using a PNA blocker, indicating that this species lacked a significant fungal microbiome that was amplified using this sequencing approach. Using a mono-association mosquito infection model, we confirmed that mosquito-derived fungal isolates colonize and for Aedes mosquitoes, support growth and development at comparable rates to their bacterial counterparts. Strikingly, native bacterial taxa isolated from mosquitoes impeded the colonization of symbiotic fungi in Ae. aegypti suggesting interkingdom interactions shape fungal microbiome communities. Conclusion Collectively, this study adds to our understanding of the fungal microbiome of different mosquito species, that these fungal microbes support growth and development, and highlights that microbial interactions underpin fungal colonization of these medically relevent species.

The establishment of effective antiviral responses within host cells is intricately related to their metabolic status, shedding light on immunometabolism. In this study, we investigated the hypothesis that cellular reliance on glutamine metabolism contributes to the development of a potent antiviral response. We evaluated the antiviral response in the presence or absence of L-glutamine in the culture medium, revealing a bivalent response hinging on cellular metabolism. While certain interferon-stimulated genes (ISGs) exhibited higher expression in an oxidative phosphorylation (OXPHOS)-dependent manner, others were surprisingly upregulated in a glycolytic-dependent manner. This metabolic dichotomy was influenced in part by variations in IFN-β expression. We initially demonstrated that the presence of L-glutamine induced an enhancement of OXPHOS in A549 cells. Furthermore, in cells either stimulated poly:IC or infected with Dengue Virus and Zika Virus, a marked increase in ISGs expression was observed in a dose-dependent manner with L-glutamine supplementation. Interestingly, our findings unveiled a metabolic dependency in the expression of specific ISGs. In particular, genes such as ISG54, ISG12 and ISG15 exhibited heightened expression in cells cultured with L-glutamine, corresponding to higher OXPHOS rates and IFN-β signaling. Conversely, the expression of viperin and 2’-5’-oligoadenylate synthetase 1 was inversely related to L-glutamine concentration, suggesting a glycolysis-dependent regulation, confirmed by inhibition experiments. This study highlights the intricate interplay between cellular metabolism, especially glutaminergic and glycolytic, and the establishment of the canonical antiviral response characterized by the expression of antiviral effectors, potentially paving the way for novel strategies to modulate antiviral responses through metabolic interventions.

In the last decade and a half, the world has experienced the outbreak of a range of viruses such as COVID-19, H1N1, flu, Ebola, Zika Virus, Middle East Respiratory Syndrome (MERS), Measles, and West Nile Virus, just to name a few. During these virus outbreaks, the usage and effectiveness of social media platforms increased significantly as such platforms served as virtual communities, enabling their users to share and exchange information, news, perspectives, opinions, ideas, and comments related to the outbreaks. Analysis of this Big Data of conversations related to virus outbreaks using concepts of Natural Language Processing such as Topic Modeling has attracted the attention of researchers from different disciplines such as Healthcare, Epidemiology, Data Science, Medicine, and Computer Science. The recent outbreak of the MPox virus has resulted in a tremendous increase in the usage of Twitter. Prior works in this field have primarily focused on the sentiment analysis and content analysis of these Tweets, and the few works that have focused on topic modeling have multiple limitations. This paper aims to address this research gap and makes two scientific contributions to this field. First, it presents the results of performing Topic Modeling on 601,432 Tweets about the 2022 Mpox outbreak, which were posted on Twitter between May 7, 2022, and March 3, 2023. The results indicate that the conversations on Twitter related to Mpox during this time range may be broadly categorized into four distinct themes - Views and Perspectives about MPox, Updates on Cases and Investigations about Mpox, MPox and the LGBTQIA+ Community, and MPox and COVID-19. Second, the paper presents the findings from the analysis of these Tweets. The results show that the theme that was most popular on Twitter (in terms of the number of Tweets posted) during this time range was - Views and Perspectives about MPox. It is followed by the theme of MPox and the LGBTQIA+ Community, which is followed by the themes of MPox and COVID-19 and Updates on Cases and Investigations about Mpox, respectively. Finally, a comparison with prior works in this field is also presented to highlight the novelty and significance of this research work.

The recent COVID-19 pandemic shows the critical need for novel broad spectrum antiviral agents. Scorpion venoms are known to contain highly bioactive peptides, several of which have demonstrated strong antiviral activity against a range of viruses. We have generated the first annotated reference transcriptome for the Androctonus amoreuxi venom gland and used high performance liquid chromatography, transcriptome mining, circular dichroism and mass spectrometric analysis to purify and characterize fourteen new venom peptides. Selected peptides were tested for binding to the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein and inhibition of the spike RBD – human angiotensin-converting enzyme 2 (hACE2) interaction using surface plasmon resonance-based assays. Seven peptides showed dose-dependent inhibitory effects. The most active peptide was synthesized using solid phase peptide synthesis and tested for its antiviral activity against SARS-CoV-2 (Lineage B.1.1.7). On exposure of replication-competent SARS-CoV-2 to the synthetic peptide, we observed a two log10 PFU/mL reduction at sub-micromolar concentrations of the peptide compared to virus exposed to medium alone. Our results show that scorpion venom peptides can inhibit the SARS-CoV-2 spike RBD – hACE2 interaction, exhibit anti-SARS-CoV-2 activity through other unexplored modes of 3 actions and represent excellent scaffolds for design of novel anti-SARS-CoV-2 constrained peptides.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging porcine intestinal coronavirus that can cause acute diarrhea, vomiting, rapid weight loss, and high mortality in newborn piglets. Cholesterol 25-hydroxylase (CH25H) is a molecular mediator of innate antiviral immunity and converts cholesterol to 25-hydroxycholesterol (25HC). Previous studies have reported that CH25H and 25HC have an antiviral effect against multiple viruses. However, it is still uncertain about the interplay between SADS-CoV infection and CH25H or 25HC. Here, we found that CH25H and its enzymatic products 25HC restrained SADS-CoV replication by blocking membrane fusion. Our results show that CH25H was upregulated by SADS-CoV infection in vitro and in vivo, and that it was an IFN-stimulated gene in IPI-2I cells. Moreover, CH25H and CH25H mutants lacking catalytic activity can inhibit SADS-CoV replication. Further, 25HC significantly suppressed SADS-CoV infection by inhibiting virus entry. Notably, we confirmed that CH25H and 25HC blocked SADS-CoV spike protein-mediated membrane fusion. Our data provide a possible antiviral therapy against SADS-CoV and other conceivable emerging coronaviruses in the future.

Infectious disease forecasting has been a key focus and proved to be crucial in controlling epidemic. A recent trend is to develop forecast-ing models based on graph neural networks (GNNs). However, existing GNN-based methods suffer from two key limitations: (1) Current models broaden receptive fields by scaling the depth of GNNs, which is insuffi-cient to preserve the semantics of long-range connectivity between distant but epidemic related areas. (2) Previous approaches model epidemics within single spatial scale, while ignoring the multi-scale epidemic pat-terns derived from different scales. To address these deficiencies, we devise the Multi-scale Spatio-temporal Graph Neural Network (MSGNN) based on an innovative multi-scale view. To be specific, in the proposed MSGNN model, we first devise a novel graph learning module, which directly captures long-range connectivity from trans-regional epidemic signals and integrates them into a multi-scale graph. Based on the learned multi-scale graph, we utilize a newly designed graph convolution module to exploit multi-scale epidemic patterns. This module allows us to facilitate multi-scale epidemic modeling by mining both scale-shared and scale-specific pat-terns. Experimental results on forecasting new cases of COVID-19 in United State demonstrate the superiority of our method over state-of-arts. Further analyses and visualization also show that MSGNN offers not only accurate, but also robust and interpretable forecasting result.