The current recommendation to avoid non-steroidal anti-inflammatory drugs (NSAIDs) in management of dengue virus disease is, as it was adopted earlier in COVID-19, scientifically considered of very low to low certainty. In this perspective I argue, for the first time globally, basing on real-life practice that NSAIDs might be lifesaving in early management of dengue virus disease as I originally argued and proved rightful in COVID-19. Moreover, I also argue that the Egyptian immune-modulatory Kelleni’s protocol, including nitazoxanide as an integral component, can be safely and effectively used to early manage COVID-19, human metapneumovirus, dengue virus disease, Zika virus separate or co-infections similar to what has already been experienced and published with separate or conjoined infections with SARS CoV-2, influenza, RSV and Norwalk viruses without the need of prior serological or molecular investigations to determine the causative agent.

Interferon Stimulated Genes (ISGs) play key roles in the control of viral replication and dissemination. Understanding this dynamic relationship between the pathogen and host is critical to our understanding of viral life-cycles and development of potential novel anti-viral strategies. Traditionally, plasmid based exogenous prompter driven expression of ISGs has been used to investigate anti-viral ISG function, however there are deficiencies in this approach. To overcome this, we investigated the utility of CRISPR activation (CRISPRa), which allows for targeted transcriptional activation of a gene from its endogenous promoter. Using the CRISPRa-SAM system to induce targeted expression of a panel of anti-viral ISGs we showed robust induction of mRNA and protein expression. We then employed our CRISPRa-SAM ISG panel in several antiviral screen formats to test for the ability of ISGs to prevent viral induced cytopathic cell death (CPE) and replication of Dengue Virus (DENV), Zika Virus (ZIKV), West Nile Virus Kunjin (WNV KUNV ), Hepatitis A Virus (HAV) and Human Coronavirus 229E (HCoV-229E). Our CRISPRa approach confirmed the anti-viral activity of ISGs like IFI6, IFNβ and IFNλ2 that prevented viral induced CPE, which was supported by high-content immunofluorescence imaging analysis. This work highlights CRISPRa as a rapid, agile, and powerful methodology to identify and characterise ISGs and viral restriction factors.

Background: To report a case of bilateral rapidly progressive visual loss in a patient with long-standing Crohn's disease treated with a TNF blocking agent for 4 months who received a second SARS-CoV-2 mRNA dose 7 weeks before onset. Case presentation A 26-year-old female with a 5-year history of Crohn’s disease was treated with adalimumab (Humira ® ) bimonthly from March 2021 to the end of July 2021. She received a first dose of mRNA-1273 COVID-19 vaccine (Moderna ® ) on April 14 th, 2021, and a second dose was given on May 18 th, 2021. Forty-nine days after the second dose, she complained of a central scotoma in her right eye. Her ophthalmologist noticed bilateral subretinal fluid associated with mild vasculitis on angiography. She was treated with 50 mg/day of oral prednisone and then substituted two days later by acetazolamide 250 mg/day when she was referred to an uveitis specialist in the city. Visual acuity worsened, and she was hospitalized from July 29 th to August 4 th, 2021, for bilateral panuveitis with severe vasculitis. She received five days of 500 mg/day intravenous methylprednisolone, followed by oral prednisone 1.5 mg/kg/day plus 1200 mg/day intravenous acyclovir and 100 mg/day acetylsalicylic acid. Adalimumab was switched to infliximab (Inflectra ® ). When she was first seen at Jules-Gonin Eye Hospital on November 4, 2021, the visual acuity was reduced to light perception in both eyes (OU). Fundus examination revealed bilateral honeycomb macular atrophy and multiple peripheral focal lesions with massive retinal ischemia. Optic disc atrophy was present in the OU. Ocular infections were ruled out by an anterior chamber tap puncture and multiple serologies, including tropical infections. Antiphospholipid antibodies were repeatedly negative. There were no signs of systemic involvement, and Crohn's disease was inactive. For corticosteroid sparing, weekly injections of tocilizumab were performed for one year from 6 th December 2021, and corticosteroids were progressively tapered to 7.5 mg/day in January 2023. Conclusions: This case study depicts a doomed visual fate in a young woman. Several hypotheses have been raised about the pathogenesis of the disease: either an autoimmune process with massive vascular occlusive disease, a reaction to biologic agent (adalimumab) injections or a boost of ocular inflammation after COVID-19 vaccination.

The World Health Organization (WHO) added Disease X to their shortlist of blueprint priority diseases to represent a hypothetical, unknown pathogen that could cause a future epidemic. During different virus outbreaks of the past, such as COVID-19, Influenza, Lyme Disease, and Zika virus, researchers from various disciplines utilized Google Trends to mine multimodal components of web behavior to study, investigate, and analyze the global awareness, preparedness, and response associated with these respective virus outbreaks. As the world prepares for Disease X, a dataset on web behavior related to Disease X would be crucial to contribute towards the timely advancement of research in this field. Furthermore, none of the prior works in this field have focused on the development of a dataset to compile relevant web behavior data, which would help to prepare for Disease X. To address these research challenges, this work presents a dataset of web behavior related to Disease X, which emerged from different geographic regions of the world, between February 2018 to August 2023. Specifically, this dataset presents the search interests related to Disease X from 94 geographic regions. These regions were chosen for data mining as these regions recorded significant search interests related to Disease X during this timeframe. The dataset was developed by collecting data using Google Trends. The relevant search interests for all these regions for each month in this time range are available in this dataset. This paper also discusses the compliance of this dataset with the FAIR principles of scientific data management. Finally, a brief analysis of specific features of this dataset is presented to uphold the applicability, relevance, and usefulness of this dataset for the investigation of different research questions in the interrelated fields of Big Data, Data Mining, Healthcare, Epidemiology, and Data Analysis.

Our growing ability to tailor healthcare to the needs of individuals has the potential to transform clinical treatment. However, the measurement of multiple biomarkers to inform clinical decisions requires rapid, effective, and affordable diagnostics. Chronic diseases and rapidly evolving pathogens in a larger population have also escalated the need for improved diagnostic capabilities. Current chemical diagnostics are often performed in centralised facilities and are still dependent on multiple steps, molecular labelling, and detailed analysis causing the result turnaround time to be over hours and days. Rapid diagnostic kits based on lateral flow devices can return results quickly but are only capable of detecting a handful of pathogens or markers. Herein we present the use of disposable plasmonics as a platform for low-cost label-free optical biosensing with multiplexing capabilities, eliminating the need for flow systems often required in current optical biosensors. We demonstrate the biosensing capabilities of a system that uses chiroptical measurements as well as plasmonic enhanced fluorescence from metafilms with shuriken shaped nanoindentations. We also perform the detection of SARS-CoV-2 in complex solutions as an early developmental milestone towards high-throughput, single-step, diagnostic kits for differential diagnosis of multiple respiratory viruses and any other emerging diagnostic needs. Diagnostics based on this platform which we term Disposable Plasmonics Assays, would be suitable for low-cost screening of multiple pathogens or biomarkers in a near point of care setting.

The development of sensitive, fast, and more reliable diagnostic methods to prevent and control the outbreak of life-threatening pandemic infectious diseases, such as COVID-19, according to the WHO recommendations, is always in priority and necessity. Optical diagnostic methods, with their progress in technology, are central to most applications and now, in the health field, are an alternative to conventional methods because of their great sensitivity and capability utilized for SARS-CoV-2 detection such as generally applied PCR technology which is time-consuming, effortful, and has limited availability in resource-limited situations. Herein, we reviewed various optical methods engaged for virus detection such as fluorescence-based techniques, Raman spectroscopy (RS), Plasmonic methods (e.g., SPR, LSPR, SEF, and SERS), ATR-FTIR, super-resolution microscopy (SRM), and some integrated platforms like waveguides and MIP-based biosensors for the development of portable, sensitive, specific, and low-cost POC apparatus for the rapid virus detection.

Data on SARS-CoV-2 infection in pregnancy and infancy has accumulated throughout the course of the pandemic. However, limited information is available from countries in sub-Saharan Africa (SSA). Evidence regarding asymptomatic SARS-CoV-2 infection and adverse birth outcomes are also scarce in these countries. The pregnant woman and infant COVID in Africa study (PeriCOVID Africa) is a South-South-North partnership involving hospitals and health centres in five countries: Malawi, Uganda, Mozambique, The Gambia, and Kenya. The study leveraged data from three ongoing prospective cohort studies: Preparing for Group B Streptococcal Vaccines (GBS PREPARE), SARS-CoV-2 infection and COVID-19 in women and their infants in Kampala and Mukono (COMAC) and Pregnancy Care Integrating Translational Science Everywhere (PRECISE). In this paper we describe the seroepidemiology of SARS-CoV-2 infection in pregnant women enrolled in sites in Uganda and Malawi, and the impact of SARS-CoV-2 infection on pregnancy and infant outcomes. The PeriCOVID study is a prospective mother-infant cohort study that recruited pregnant women at any gestation antenatally or on the day of delivery. A nasopharyngeal swab was taken from mothers at enrolment for RT-PCR confirmation of SARS-CoV-2 infection, and maternal and cord blood samples were tested for SARS-CoV-2 antibodies using Wantai and Euroimmune ELISA. The primary outcome was seroprevalence of SARS-CoV-2 antibodies in maternal blood, reported as the proportion of seropositive women by study site and wave of COVID-19 within each country. Placental transfer of antibodies was described using the geometric mean ratio (GMR). We also estimated the proportion of asymptomatic or subclinical COVID-19 infections in pregnant women using serological testing and collected adverse pregnancy and infancy outcomes (e.g. still-birth, prematurity, maternal or infant death). In total, 1379 women were enrolled, giving birth to 1387 infants. Overall, 63% of pregnant women had a SARS-CoV-2 positive serology. Over subsequent waves (delta and omicron), in the absence of vaccination, seropositivity rose from 20% to over 80%. The placental transfer GMR was 1.7, indicating active placental transfer of anti-spike IgG. There was no association between SARS-CoV-2 antibody positivity and adverse pregnancy or infancy outcomes. This study describes the increasing prevalence of SARS CoV-2 antibodies in pregnant woman in Uganda and Malawi across waves of SARS-CoV-2 infection. Our study adds to existing evidence that suggests under-reporting of infection if based solely on cases with clinical disease, or a positive RT-PCR for SARS-CoV-2, as most of the women in our study had asymptomatic infections and did not seek medical care. This has implications for screening in subsequent outbreaks and pandemics where protection of pregnant women and effect of infection in pregnancy on the infant are unknown.

Zika virus (ZIKV) infection during pregnancy poses significant threats to maternal and fetal health, leading to intrauterine fetal demise and severe developmental malformations that constitute congenital Zika syndrome (CZS). As such, the development of a safe and effective ZIKV vaccine is a critical public health priority. However, the safety and efficacy of such a vaccine during pregnancy remain uncertain. Historically, the conduct of clinical trials in pregnant women has been challenging. Therefore, clinically relevant animal pregnancy models are in high demand for testing vaccine efficacy. We previously reported that a marmoset pregnancy model of ZIKV infection consistently demonstrated vertical transmission from mother to fetus during pregnancy. Using this marmoset model, we also showed that vertical transmission could be prevented by pre-pregnancy vaccination with Zika purified inactivated virus (ZPIV) vaccine. Here, we further examined the efficacy of ZPIV vaccination during pregnancy. Vaccination during pregnancy elicited virus neutralizing antibody responses that were comparable to those elicited by pre-pregnancy vaccination. Vaccination also prevented vertical transmission of ZIKV during pregnancy, without causing adverse effects. These results provide key insights into the safety and protective efficacy of ZPIV vaccination during pregnancy, an important advance in preparedness for future ZIKV outbreaks.

Zika virus (ZIKV) is a significant threat to pregnant women and fetuses as it can cause severe birth defects and congenital neurodevelopmental disorders, referred to as congenital Zika syndrome (CZS). Thus, a safe and effective ZIKV vaccine for pregnant women to prevent in utero ZIKV infection is of utmost importance. Murine models of ZIKV infection are limited by the fact that immunocompetent mice are resistant to ZIKV infection. As such, interferon-deficient mice have been used in some preclinical studies to test the efficacy of ZIKV vaccine candidates against lethal virus challenge. However, interferon-deficient mouse models have limitations in assessing the immunogenicity of vaccines, necessitating the use of immunocompetent mouse pregnancy models. Using the human stat2 knock-in (hSTAT2KI) mouse pregnancy model, we show that vaccination with a purified formalin-inactivated Zika virus vaccine (ZPIV) prior to pregnancy successfully prevented vertical transmission. In addition, maternal immunity protected offspring against postnatal challenge. Furthermore, passive transfer of human IgG purified from hyper-immune sera of ZPIV vaccinees prevented maternal and fetal ZIKV infection, providing strong evidence that the neutralizing antibody response may serve as a meaningful correlate of protection.

Background: Coastal Kenya has experienced repeated outbreaks of Chikungunya (CHIKV) and Dengue (DENV) viruses mediated by competent Aedes aegypti mosquito populations . These mosquitoes harbor insect specific viruses (ISVs), some of which can prevent arboviral transmission . However, there has been no systematic molecular entomological surveillance in coastal Kenya and the diversity of viruses in local Aedes aegypti populations remains largely unknown. Methods: : To obtain a snapshot of the Aedes aegypti viromes from coastal Kenya, we took advantage of a cross-sectional survey of mosquitoes to determine the prevalence of Zika virus. We collected adult mosquitoes using lured Biogent’s sentinel traps at 16 different localities along the Kenyan coast between May to September 2017. Pools of 20 female Aedes aegypti mosquitoes were generated following grouping by morphological characteristics. Presence of arboviruses in the mosquito pools was determined using virus-specific and genera-specific primers with real-time PCR. Metagenomic next generation sequencing (mNGS) on Illumina Miseq and analysis was used to characterize the virome. Results: : A total of 16,520 female Aedes aegypti grouped into 826 pools were analysed. Flaviviruses were detected in 69/826 (8.4%) pools by real time PCR. Sequencing generated 8,459/971,754 (0.87%) clean reads that were taxonomically assigned to 16 and 28 viral families and species, respectively. The family Phenuiviridae represented by Phasi Charoen-like phasivirus (PCLV) species was the most abundant, detected in 64/73 (87%) mosquito pools. No pathogenic viruses were identified by mNGS. Phylogenetic analysis revealed local PCLV and Cell fusing agent virus (CFAV) were distinct from global sequences. Conclusions: : Our data provides information about virus diversity and composition of the Aedes aegypti mosquitoes from coastal Kenya and contributes to the body of knowledge of the Aedes aegypti virome. To the best of our knowledge, this is the first study to provide this information from this region.

Serosurveys suggest widespread dengue virus (DENV) transmission in Africa but there is limited information on the temporal patterns of exposure. Here, we estimated the prevalence and incidence of DENV infections in coastal Kenya over a 20-year period (1998–2018). Sera (n=8038) obtained annually from a longitudinal cohort of 1847 children aged 15 years and below were screened for anti-DENV IgG antibodies. Anti-DENV IgG seroprevalence increased with age and peaked during outbreak years. Among 1354 children who were seronegative at recruitment, we observed an overall incidence (seroconversion) rate of 129.5 (95% CI 118.7–141.4) DENV infections per 1000 person-years. The highest incidence was observed in 2013 at 520 infections per 1000 person-years (95% CI 443.6–610.2) coinciding with a large DENV outbreak in coastal Kenya. Our data suggest long-term DENV exposure among children in coastal Kenya highlighting an urgent need for clinical surveillance to determine the associated health burden in this setting.

Coronavirus disease-2019 (COVID-19) was seen as a respiratory disease, however, an increasing number of reports indicated that the spike protein could also be the cause of the long-term post-infectious conditions known as Long-COVID characterized by a group of unresponsive idiopathic severe neuro, cardio-vascular disorders, including strokes, cardiopathies, neuralgias, fibromyalgia, and Parkinson's like-disease. Different lines of pieces of evidence confirmed that the spike protein that can be found on the surface of the SARS-CoV-2 virus latches onto angiotensin-converting enzyme 2 (ACE2) receptors located on target cells. The RNA genome of coronaviruses, which, has a median length of 29 kb and is the longest among all RNA viruses, is comprised of six to ten open reading frames (ORFs) that are responsible for encoding both the replicase and structural proteins for the virus. Each of the components of the viral genome is packaged into a helical nucleocapsid that is surrounded by a lipid bilayer. The viral envelope of coronaviruses is typically made up of three proteins that include the membrane protein (M), the envelope protein (E), and the spike protein (S). The spike protein not only facilitates the virus entry into healthy cells, which is the first step in infection but also promote profound damage to different organs and tissues leading to severe impairments and long-term disabilities. Here, we discussed the pervasive mechanism that spikes mRNA adopted to alter multipotent and pluripotent stem cell (SCs) genomes and the acquired disability of generating an infinite number of affected clonal cells. This stance is based on the molecular and evolutionary aspects obtained from retrotransposons-retrotransposition in mammalians and humans that documented the frequent integration of mRNA molecules into genomes and thus into DNA. Retrotransposition is the molecular process in which transcribed and spliced mRNAs are accidentally reverse-transcribed and inserted into new genomic positions to form a retrogene. Sequence-specific traits of mRNA clearly showed long interspersed element-1 (LINE-1 or L1) to confirm the retrotransposition, considered the most abundant autonomously active retrotransposons in the human genome. In mammals, L1 retrotransposons drive retrotransposition and are composed of long terminal repeats (LTRs) and non-LTR retrotransposons (mainly long interspersed nuclear elements or LINEs); specifically, the LTR-mediated retrocopies are immediately cotranscribed with their flanking LTR retrotransposons. In response to retrotransposons transposition, stem cells (SCs) employ a number of silencing mechanisms, such as DNA methylation and histone modification. This manuscript theorizes the expression patterns, functions, and regulation of mRNA Spike protein imprinted by SCs retrotransposons which generate unlimited lines of affected cell progenies and tissues as the main condition of untreatable Spike-related inflammatory conditions.

The inflammasome is a large multiprotein complex that assembles in the cell cytoplasm in response to stress or pathogenic infection. Its primary function is to defend the cell and promote the secretion of pro-inflammatory cytokines, including IL-1β and IL-18. It was shown that in immortalized bone marrow derived macrophages (iBMDMs) inflammasome assembly is dependent on the deacetylase HDAC6 and the aggresome processing pathway (APP), a cellular pathway involved in the disposal of misfolded proteins. Here we used primary BMDMs from mice in which HDAC6 is ablated or impaired and found that inflammasome activation was largely normal. We also used human peripheral blood mononuclear cells and monocytes cell lines expressing a synthetic protein blocking HDAC6-ubiquitin interaction and impairing the APP and found that inflammasome activation was moderately affected. Finally, we used a novel HDAC6 degrader and showed that inflammasome activation was partially impaired in human macrophage cell lines with depleted HDAC6. Our results therefore show that HDAC6 importance in inflammasome activation is context dependent.

Background: As a highly popular choice for a household pet worldwide, domestic cats may carry pathogens and antimicrobial resistance genes (ARGs) that pose severe risks to human health. However, our knowledge about pathogens and ARGs carried by cats remains limited. This large-scale genomic surveillance study is designed to identify viruses and bacteria as well as ARGs that may pose risks to animal and human health in the upper respiratory tracts of domestic cats in China. Results: : Meta-transcriptome sequencing of 1454 oropharyngeal-nasal swabs collected from domestic cats in different living conditions in 22 Chinese provinces identified 371 viral species that can affect the health of animals and humans. A total of 24 viral species that had been known to cause infections in cats were detected. Among them, felis catus gammaherpesvirus 1, feline dependoparvovirus, feline foamy virus, fesavirus 4, puma lentivirus, and puma lentivirus-14 were detected in Chinese cats for the first time. Our study also identified the presence of nucleic acids from viruses associated with other animal species commonly found in human daily lives, including poultry, birds, pigs, cattle and other ruminants, minks, rabbits, and dogs. Notably, we identified many well-known viruses associated with public health concerns, including SARS-related coronavirus, monkeypox virus, and Zika virus (ZIKV). For the first time we found cats could be experimentally infected with and transmit ZIKV via mosquito bites, indicating a potential role of domestic cats in the maintenance and spread of the virus. We also identified a substantial number of bacterial species associated with public health, including the ESKAPE pathogens ( Enterococcus faecium , Staphylococcus aureus , Klebsiella pneumoniae , Acinetobacter baumannii , Pseudomonas aeruginosa , and Enterobacter spp.) which showed a broad detection range. Additionally, a plentiful amount of ARGs, including those conferring resistance to the last resort antibiotics (carbapenems [ bla NDM , bla OXA , bla VIM ], colistin [ mcr ], and high-level tigecycline [ tetX3 , tetX4 , tetX5 , tetX6 ]), were also detected broadly. Conclusions: : This study indicate that domestic cats may act as a role in the worldwide emergence and increasing of zoonotic diseases and antimicrobial resistance. It is important to monitoring domestic cats within the One Health approach to control these critical global public health issues.

Abstract Nucleic acid tests are essential for the accurate diagnosis and control of infectious diseases. However, current assays are not easily scalable for a large population, due to the requirement of laboratory settings or special equipment. Here, we developed an integrated box for instant nucleic acid screening (BINAS) which fully integrates nucleic acid release, amplification, and results visualization for self-service standalone test. Importantly, operation of BINAS runs on a novel gamepad-like interface, which allows deployment of BINAS in home settings and operation by users without any prior professional training. Performance of BINAS is empowered by a RNA extraction-free sample inactivation process and a newly developed Nest-RPA chemistry and exhibits sensitivity comparable to RT-qPCR with high specificity for SARS-Cov-2 RNA in a reaction time of 30 minutes directly from fresh swab sample to results. These innovations make BINAS a novel platform for a convenient, accurate, and deployable point-of-care testing scheme.

ABSTRACT Endocytosis, or internalization through endosomes is a major cell entry mechanism used by respiratory viruses. Phosphoinositide 5-kinase (PIKfyve) is a critical enzyme for the synthesis of Phosphatidylinositol (3,5)biphosphate (PtdIns(3,5)P2), and has been implicated in virus trafficking via the endocytic pathway. In fact, antiviral effects of PIKfyve inhibitors against SARS-CoV-2 and Ebola have been reported, but there is little evidence regarding other respiratory viruses. In this study we demonstrated the antiviral effects of PIKfyve inhibitors on influenza virus and respiratory syncytial virus in vitro and in vivo . PIKfyve inhibitors, Apilimod mesylate (AM) and YM201636 concentration-dependently inhibited several influenza strains in a MDCK cell-cytopathic assay. AM also reduced the viral load and cytokine release, whilst improving the cell integrity of human nasal air liquid interface cultured epithelium infected with influenza PR8. In PR8-infected mice, AM (2mg/ml), when intranasally treated, exhibited significant reduction of viral load and inflammation and inhibited weight loss caused by influenza infection, with effects being similar to oral oseltamivir (10 mg/kg). In addition, AM demonstrated anti-viral effects in RSV A2 infected human nasal epithelium in vitro and mouse in vivo , with equivalent effect to that of ribavirin. AM also showed anti-viral effects against human rhinovirus and seasonal coronavirus in vitro . Thus, PIKfyve is found to be involved in influenza and RSV infection, and PIKfyve inhibitor is a promising molecule for pan-viral approach against respiratory viruses.

Molecular tools for modulating transgene expression in Aedes aegypti are few. Here we demonstrate that adjustments to the AePUb promoter length can alter expression levels of two reporter proteins in Ae. aegypti cell culture and in mosquitoes. This provides a simple means for increasing or decreasing expression of a gene of interest and easy translation from cells to whole insects.

Chikungunya, Dengue, and Zika viruses are endemic to Brazil, with a high annual incidence rate. It is a serious public health issue and is responsible for outbreaks and epidemics worldwide. As COVID-19 emerged as a pandemic, the diagnosis of arboviruses was underestimated. Hence, this study aimed to characterize the sociodemographic and clinical aspects of arboviruses in the con-text of the COVID-19 pandemic in the state of Maranhão, Brazil. Sociodemographic and clinical data were obtained by applying questionnaires to 179 patients treated at health units in São José de Ribamar, Paço do Lumiar, Raposa, Santa Inês, and Vargem Grande municipalities. Serologi-cal and polymerase chain reaction analyses were performed do detect Chikungunya, Zika, Den-gue, and COVID-19 infections. The Maranhão state registered 706 probable cases of Chikungunya from 2019 to 2020. In 2020, the cases were reduced by 74.8% compared to those reported in 2019. The test results were analyzed separately. Here, 46.3% of the Chikungunya virus test results were negative, and 36.8% were positive. For the Zika virus tests, most (79.9%) were positive, as well as the DENV1, DENV2, DENV3, and DENV4 tests (88.3%, 88.3%, 89.4%, and 65.9%, respectively). For the COVID-19 tests, of the 146 individuals tested, 52.7% were reactive when considering the anti-S test, and 17.8% were positive when considering the anti-N test. Fever and myalgia were the symptoms most frequently reported by patients. Our results emphasize the importance of contin-uous epidemiological surveillance of arboviruses in northeastern Brazil, and simultaneous test-ing for Chikungunya, Dengue, and Zika viruses among suspected patients.

Zika virus (ZIKV) outbreak caused one of the most significant medical emergencies in the Americas due to associated microcephaly in newborns. To evaluate the impact of ZIKV infection on neuronal cells over time, we retrieved gene expression data from several ZIKV-infected samples obtained at different time point post-infection (pi). Differential gene expression analysis was applied at each time point, with more differentially expressed genes (DEG) identified at 72h pi. There were 5 DEGs (PLA2G2F, TMEM71, PKD1L2, UBD, and TNFAIP3 genes) across all timepoints, which clearly distinguished between infected and healthy samples. The highest expression levels of all five genes were identified at 72h pi. Taken together, our results indicate that ZIKV infection greatly impacts human neural cells at early times of infection, with peak perturbation observed at 72h pi. Our analysis revealed that all five DEGs, in samples of ZIKV-infected human neural stem cells, remained highly upregulated across the timepoints evaluated. Moreover, despite the pronounced inflammatory host response observed throughout infection, the impact of ZIKV is variable over time. Finally, the five DEGs identified herein play prominent roles in infection, and could serve to guide future investigations into virus-host interaction, as well as constitute targets for therapeutic drug development.

This paper makes two key contributions. First, it argues that highly specialized rare content classifiers trained on small data typically have limited exposure to the richness and topical diversity of the negative class (dubbed anticontent) as observed in the wild. As a result, these classifiers' strong performance observed on the test set may not translate into real-world settings. In the context of COVID-19 misinformation detection, we conduct an in-the-wild audit of multiple datasets and demonstrate that models trained with several prominently cited recent datasets are vulnerable to anticontent when evaluated in the wild. Second, we present a novel active learning pipeline that requires zero manual annotation and iteratively augments the training data with challenging anticontent, robustifying these classifiers.

Traditional vaccines are produced by using weakened or inactivated forms of disease-causing pathogens to produce the target antigen they are designed to protect against. Messenger RNA vaccines are a class of vaccines that employ a minute segment of genetic material, known as messenger RNA (mRNA), which contains directives for the cells in the body to generate a particular protein. This genetic material is synthesized in the laboratory and packaged into a lipid nanoparticle, which protects and helps it enter cells for further protein synthesis. During vaccination with mRNA vaccine, the lipid nanoparticles containing the mRNA are injected into the muscle of vaccinees. Once inside the cells, the mRNA instructs the cells to produce a protein which is then displayed on the surface of the cell, triggering an immune response. During this, the immune system recognizes the displayed protein as foreign and mounts a defense by producing antibodies and activating immune cells to target and eliminate the protein. Furthermore, these immune responses generate a memory cell, facilitating the immune system to promptly react in case of encountering the authentic pathogen as an infection in the future. The mRNA vaccines are flexible and the sequence can be easily synthesized in the lab based on the genetic information of the target pathogen. Additionally, mRNA vaccines can be developed for new strains or variants of the target disease easily. This was particularly evident during the COVID-19 pandemic, where mRNA vaccines like the Pfizer-BioNTech and Moderna vaccines were developed and authorized for emergency use within a year. But currently, available mRNA vaccines require extensive cold chain, antigen delivery, potential immune response variability optimization, and sophisticated manufacturing process. The efforts to explore next-generation mRNA vaccine development are aimed to further improve the effectiveness, stability, and delivery methods. One focus of research has been to enhance the stability of mRNA vaccines, particularly temperature sensitivity which makes storage and distribution easier, particularly in regions with limited access to cold chain infrastructure. Self-amplifying mRNA vaccines, on the other hand, are designed to generate multiple copies of the mRNA within cells which potentially leads to a higher production of the target protein, resulting in a stronger immune response. Additionally, studies are exploring new delivery systems to improve the target and efficiency of mRNA vaccines using specialized nanoparticles and liposomes to specifically deliver mRNA to certain cell types or immune cells. Another area of interest is the development of combination vaccines, where multiple mRNA sequences are included in a single vaccine protecting against multiple diseases targeting strains or variants of a particular pathogen simultaneously. While current mRNA vaccines are administered via intramuscular injection, studies are underway to deliver directly into the skin offering enhanced immune response and the ability to use smaller vaccine doses.

ABSTRACT Bacille Calmette-Guérin (BCG) vaccination can confer non-specific protection against heterologous pathogens. However, the underlying mechanisms remain mysterious. Here, we show that mice immunized intravenously with BCG exhibited reduced weight loss and/or improved viral clearance when challenged with SARS-CoV-2 and influenza. Protection was first evident between 14 - 21 days post vaccination, and lasted for at least 42 days. Remarkably, BCG induced a biphasic innate response in the lung, initially at day 1 and a subsequent prolonged phase starting at ∼15 days post vaccination, and robust antigen-specific Th1 responses. MyD88-dependent TLR signaling was essential for the induction of the innate and Th1 responses, and protection against SARS-CoV-2. Depletion of CD4 + T cells or IFN-γ activity prior to infection obliterated innate activation and protection. Single cell and spatial transcriptomics revealed CD4-dependent expression of interferon-stimulated genes (ISGs) in myeloid, type II alveolar and lung epithelial cells. Thus, BCG elicits “integrated organ immunity” where CD4+ T cells act on local myeloid and epithelial cells to imprint prolonged antiviral innate resistance.

Using nationally representative data from India, we document the first survey-based evidence of the unintended consequences of lockdown on neonatal mortality in a developing country. Event-study shows neonatal mortality significantly increased during the first nationwide lockdown and became insignificant one-month later. The difference-indifference estimates show neonatal mortality increased to 47 from 30 per 1,000 births during the lockdown. Negative in-utero exposure, forgone healthcare (through service interruption and avoidance), and delaying vaccinations are crucial impact mechanisms. Our findings stimulate the debate on the efficacy of strict lockdown, its duration, and missing policy directives in resource-poor countries, particularly for the care-dependent population.

Reverse genetic systems have been used to introduce heterologous sequences into the rotavirus segmented double-stranded (ds)RNA genome, enabling the generation of recombinant viruses that express foreign proteins and possibly serve as vaccine vectors. Notably, insertion of SARS-CoV-2 sequences into the segment 7 (NSP3) RNA of simian SA11 rotavirus was previously shown to result in the production of recombinant viruses that efficiently expressed the N-terminal domain (NTD) and the receptor-binding domain (RBD) of the S1 region of the SARS-CoV-2 spike protein. However, efforts to generate a similar recombinant (r) SA11 virus that efficiently expressed full-length S1 were less successful. In this study, we describe modifications to the S1-coding cassette inserted in the segment 7 RNA that allowed recovery of second-generation rSA11 viruses that efficiently expressed the ∼120-kDa S1 protein. The ∼120-kDa S1 products were shown to be glycosylated, based on treatment with endoglycosidase H, which reduced the protein to a size of ∼80 kDa. Co-pulldown assays demonstrated that the ∼120-kDa S1 proteins had affinity for the human ACE2 receptor. Although all the second-generation rSA11 viruses expressed glycosylated S1 with affinity for the ACE receptor, only the S1 product of one virus (rSA11/S1f) was appropriately recognized by anti-S1 antibody, suggesting the rSA11/S1f virus expressed an authentic form of S1. Probably due to the presence of FLAG tags on their S1 signal peptides, the S1 products of the other viruses (rSA11/3fS1 and rSA11/3fS1-His) may have undergone defective glycosylation, impeding antibody binding. In summary, these results indicate that recombinant rotaviruses can serve as expression vectors of foreign glycosylated proteins, raising the possibility of generating rotavirus-based vaccines that can induce protective immune responses against enteric and mucosal viruses with glycosylated capsid components, including SARS-CoV-2.

Preventing vector-borne diseases (VBDs) mainly relies on effective vector control tools and strategies, which in turn depend on population acceptance and adherence. Inspired by the abundant recent literature on SARS-COV-2, we investigate the relationship between risk perception and preventive behaviour for selected VBDs and the extent to which risk perception is determined by social norms. We use cross-sectional data collected from 497 individuals in four regions of Guyana in 2017. We use a conditional mixed process estimator with multilevel coefficients, estimated through a Generalized Linear Model (GLM) framework, applying a simultaneous equation structure. We find robust results on malaria: risk perception was significantly influenced by the risk perception of the reference group across different definitions of the reference group, hinting at the existence of social norms. Risk perception significantly increased the likelihood of passive behaviour by 4.48%. Less clear-cut results were found for dengue. This study applies quantitative social science methods to public health issues in the context of VBDs. Our findings point to the relevance of tailoring communications on health risks for VBDs to groups defined at the intersection of socio-economic and demographic characteristics. Such tailored strategies are expected to align risk perception among reference groups and boost preventive behaviour.