Bats are natural reservoirs for several zoonotic viruses, potentially due to an enhanced capacity to control viral infection. However, the mechanisms of antiviral responses in bats are poorly defined. Here we established a Jamaican fruit bat (JFB) intestinal organoid model of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. JFB organoids were susceptible to SARS-CoV-2 infection, with increased viral RNA and subgenomic RNA detected in cell lysates and supernatants. Gene expression of type I interferons and inflammatory cytokines was induced in response to SARS-CoV-2 but not in response to TLR agonists. Interestingly, SARS-CoV-2 did not lead to cytopathic effects in JFB organoids but caused enhanced organoid growth. Proteomic analyses revealed an increase in inflammatory signaling, cell turnover, cell repair, and SARS-CoV-2 infection pathways. Collectively, our findings suggest that primary JFB intestinal epithelial cells can mount a successful antiviral interferon response and that SARS-CoV-2 infection in JFB cells induces protective regenerative pathways.

Host-based antivirals could offer broad-spectrum therapeutics and prophylactics against the constantly-mutating viruses including the currently-ravaging coronavirus, yet must target cellular vulnerabilities of viruses without grossly endangering the host. Here we show that the master lipid regulator SREBP1 couples the phospholipid scramblase TMEM41B to constitute a host “metabolism-to-manufacture” cascade that maximizes membrane supplies to support coronaviral genome replication, harboring biosynthetic enzymes including Lipin1 as druggable viral-specific-essential (VSE) host genes. Moreover, pharmacological inhibition of Lipin1, by a moonlight function of the widely-prescribed beta-blocker Propranolol, metabolically uncouples the SREBP1-TMEM41B cascade and consequently exhibits broad-spectrum antiviral effects against coronaviruses, Zika virus, and Dengue virus. The data implicate a metabolism-based antiviral strategy that is well tolerated by the host, and a potential broad-spectrum medication against current and future coronavirus diseases.

SARM1 is a central regulator of programmed axon death and is required to initiate axon self-destruction after traumatic and toxic insults to the nervous system. Abnormal activation of this axon degeneration pathway is increasingly recognized as a contributor to human neurological disease and SARM1 knockdown or inhibition has become an attractive therapeutic strategy to preserve axon loss in a variety of disorders of the peripheral and central nervous system. Despite this, it remains unknown whether Sarm1 /SARM1 is present in myelinating glia and whether it plays a role in myelination in the PNS or CNS. It is important to answer these questions to understand whether future therapies inhibiting SARM1 function may have unintended deleterious impacts on myelination. Here we show that Sarm1 mRNA is present in oligodendrocytes in zebrafish but only detectable at low levels in Schwann cells in both zebrafish and mice. We find SARM1 protein is readily detectable in murine oligodendrocytes in vitro and in vivo and activation of endogenous SARM1 in oligodendrocytes induces cell death. In contrast, SARM1 protein is not detectable in Schwann cells and satellite glia in the adult murine nervous system. Cultured Schwann cells contain negligible functional SARM1 and are insensitive to specific SARM1 activators. Using zebrafish and mouse Sarm1 mutants, we show that SARM1 is not required for initiation of myelination nor myelin sheath maintenance by oligodendrocytes and Schwann cells. Thus, strategies to inhibit SARM1 function in the nervous system to treat neurological disease are unlikely to perturb myelination in humans. Main Points SARM1 protein is detectable in oligodendrocytes but not in Schwann cells Oligodendrocytes but not Schwann cells die in response to endogenous SARM1 activation CNS nor PNS myelination, in zebrafish and mice, is hindered by loss of sarm1/Sarm1

In the 2016 Zika virus (ZIKV) pandemic, a previously unrecognized risk of birth defects surfaced in babies whose mothers were infected with Asian-lineage ZIKV during pregnancy. Less is known about the impacts of gestational African-lineage ZIKV infections. Given high human immunodeficiency virus (HIV) burdens in regions where African-lineage ZIKV circulates, we evaluated whether pregnant rhesus macaques infected with simian immunodeficiency virus (SIV) have a higher risk of African-lineage ZIKV-associated birth defects. Remarkably, in both SIV+ and SIV-animals, ZIKV infection early in the first trimester caused a high incidence (78%) of spontaneous pregnancy loss within 20 days. These findings suggest a significant risk for early pregnancy loss associated with African-lineage ZIKV infection and provide the first consistent ZIKV-associated phenotype in macaques for testing medical countermeasures.

The emergence of SARS-like coronaviruses is a multi-stage process from wildlife reservoirs to people. Here we characterize multiple drivers—landscape change, host distribution, and human exposure—associated with the risk of spillover of SARS-like coronaviruses to help inform surveillance and mitigation activities. We consider direct and indirect transmission pathways by modeling four scenarios with livestock and mammalian wildlife as potential and known reservoirs before examining how access to healthcare varies within clusters and scenarios. We found 19 clusters with differing risk factor contributions within a single country (N=9) or transboundary (N=10). High-risk areas were mainly closer (11-20%) rather than far (<1%) from healthcare. Areas far from healthcare reveal healthcare access inequalities, especially Scenario 3, which includes wild mammals as secondary hosts. China (N=2) and Indonesia (N=1) had clusters with the highest risk. Our findings can help stakeholders in land use planning integrating healthcare implementation and One Health actions.

Background Pregnant females affected with COVID-19 are reported to have poorer disease outcomes as compared to non-pregnant females of a similar age group. COVID-19 may lead to adverse changes in the placenta, which needs to be studied. Methods This is a case series of 63 pregnant women hospitalized with COVID-19 from May 2020 to February 2021.The primary outcomes were maternal death or complications. Results 63 women were studied. 83.3% of women were in the age group of 26 to 35 years. 33% women had associated comorbidities. 68.3% of women tested positive in their third trimester, 15.9% and 11% tested positive in their second and first trimesters respectively. 73% women had mild disease and 27% women required oxygen support. 3/63 women died. One woman in the second and two women in the third trimester died respectively. Histopathological examination in 13 placentae (of 19 placentae examined) were suggestive of maternal and fetal malperfusion. Conclusion Pregnant COVID-19 women may develop disease-related as well as obstetric complications.

The pandemic required efficient allocation of public resources and transforming existing ways of societal functions. To manage any crisis, governments and public health researchers exploit the information available to them in order to make informed decisions, also defined as situational awareness. Gathering situational awareness using social media has been functional to manage epidemics. Previous research focused on using discussions during periods of epidemic crises on social media platforms like Twitter, Reddit, or Facebook and developing NLP techniques to filter out relevant discussions from a huge corpus of messages and posts. Social media usage varies with internet penetration and other socioeconomic factors, which might induce disparity in analyzing discussions across different geographies. However, print media is a ubiquitous information source, irrespective of geography. Further, topics discussed in news articles are already newsworthy, while on social media newsworthiness is a product of techno-social processes. Developing this fundamental difference, we study Twitter data during the second wave in India focused on six high-population cities with varied macroeconomic factors. Through a mixture of qualitative and quantitative methods, we further analyze two Indian newspapers during the same period and compare topics from both Twitter and the newspapers to evaluate situational awareness around the second phase of COVID on each of these platforms. We conclude that factors like internet penetration and GDP in a specific city influence the discourse surrounding situational updates on social media. Thus, augmenting information from newspapers with information extracted from social media would provide a more comprehensive perspective in resource deficit cities.

Altered blood hormone and metabolite levels during and post-COVID-19 have been extensively reported. Yet, studies of gene expression at the tissue level that can help identify the causes of endocrine dysfunctions are scarce. We analyzed transcript levels of endocrine-specific genes in five endocrine organs of lethal COVID-19 cases. Overall, 116 autoptic specimens from 77 individuals (50 COVID-19 and 27 uninfected controls) were included. All samples were tested for SARS-CoV-2 genome. Investigated organs included adrenals, pancreas, ovary, thyroid and white adipose tissue (WAT). Transcript levels of 42 endocrine-specific and 3 IFN-stimulated genes (ISGs) were measured and compared between COVID-19 cases (virus-positive and virus-negative in tissue) and uninfected controls. ISG transcript levels were enhanced in tissues positive for SARS-CoV-2. Endocrine-specific genes (e.g., HSD3B2 , INS , IAPP , TSHR , FOXE1 , LEP , CRYGD ) were deregulated in COVID-19 cases in an organ-specific manner. Transcription of organ-specific genes was suppressed in virus-positive specimens of ovary, pancreas and thyroid but enhanced in adrenals. In WAT of COVID-19 cases transcription of ISGs and leptin was enhanced independently of the presence of virus. Our findings suggest that, in COVID-19, endocrine dysfunctions may arise especially when SARS-CoV-2 invades endocrine organs and that transcriptional alterations of endocrine-specific genes may contribute to endocrine manifestations.

Emergence of SARS-CoV-2 variants diminishes the efficacy of vaccines and antiviral monoclonal antibodies. Continued development of immunotherapies and vaccine immunogens resilient to viral evolution is therefore necessary. Using coldspot-guided antibody discovery, a screening approach that focuses on portions of the virus spike that are both functionally relevant and averse to change, we identified human neutralizing antibodies to highly conserved viral epitopes. Antibody fp.006 binds the fusion peptide and cross-reacts against coronaviruses of the four genera , including the nine human coronaviruses, through recognition of a conserved motif that includes the S2’ site of proteolytic cleavage. Antibody hr2.016 targets the stem helix and neutralizes SARS-CoV-2 variants. Antibody sd1.040 binds to subdomain 1, synergizes with antibody rbd.042 for neutralization and, like fp.006 and hr2.016, protects mice when present as bispecific antibody. Thus, coldspot-guided antibody discovery reveals donor-derived neutralizing antibodies that are cross-reactive with Orthocoronavirinae , including SARS-CoV-2 variants. One sentence summary Broadly cross-reactive antibodies that protect from SARS-CoV-2 variants are revealed by virus coldspot-driven discovery.

ABSTRACT Background The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that receive increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Results Applying a small RNA-based metagenomic approach to Ae. japonicus , we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of 21 nucleotide sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families, however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified a ∼1 kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1 and AejapBV1 by reverse-transcriptase PCR in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100%) in adult females, adult males and larvae. Conclusions Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus and may affect different aspects of the physiology of this vector mosquito.

Abstract Background Antibodies targeting envelope glycoproteins have been shown in some instances to enhance infection by subverting Fc receptor and complement function, or by directly inducing fusion with cellular membranes. The potential for antibody dependent enhancement (ADE) of infection raises concern that passive immunization with a therapeutic anti-viral antibody could increase risk of disease. As part of the nonclinical package characterizing the risk profile of the SARS-CoV-2 neutralizing monoclonal antibody bamlanivimab, studies were conducted to evaluate the potential for ADE of infection in vitro and in a non-human primate model of COVID-19. Methods In vitro assays were performed in primary human macrophage, Raji, or THP-1 cells exposed to SARS-CoV-2 in the presence of bamlanivimab ranging from approximately IC50 to more than 100-fold above or below the IC50. Samples were evaluated for demonstration of productive viral infection. Bamlanivimab binding to C1q and FcR were quantified, and activity was studied by cell-based assays. In vivo studies were performed in African green monkeys (AGM) infected with SARS-CoV-2 virus following sub-saturating or saturating doses of bamlanivimab or IgG control. Viral loads, clinical pathology, and histology endpoints were assessed to determine if bamlanivimab enhanced SARS-CoV-2 replication or clinical illness. Mixed model repeated measures were used to evaluate virology statistics. Results Bamlanivimab did not increase viral RNA production in FcγR-expressing cell lines, despite demonstration of effector function. No significant differences were found among the AGM groups in terms of weight, temperature, or food intake. Treatment with bamlanivimab reduced viral loads in nasal and oral swabs and BAL fluid relative to control groups. Viral antigen was not detected in lung tissue from animals treated with the highest dose of bamlanivimab. Microscopic findings along with decreases in viral loads in bamlanivimab-treated animals indicated that ADE of disease was not observed in this study. Conclusions Sub-saturating doses of bamlanivimab treatment do not induce ADE of SARSCoV2 infection in either in vitro or an AGM model of infection. Findings suggest that high affinity monoclonal antibodies pose a low risk of mediating ADE in patients and further supports their safety profile as a treatment of COVID-19 disease.

Background: Understanding mortality burden associated with communicable diseases is key to informing resource allocation, disease prevention and control efforts, and evaluating public health interventions. We quantified excess mortality among people notified with communicable diseases in Victoria, Australia. Methods: Cases of communicable disease notified in Victoria between 1 January 1991–31 December 2021 were linked to the death registry. Informational gain obtained through linkage and 30-day case fatality rates were calculated for each disease. Standardised mortality ratios (SMR) and 95% confidence intervals were calculated up to a year following illness onset. Findings: There were 1,032,619 cases and 5,985 (0.58%) died ≤30 days of illness onset. Diseases with high 7-day SMR signifying excess mortality included invasive pneumococcal disease (167.7, 95% CI 153.4–182.7); listeriosis (166.2, 95% CI 121.2–218.3); invasive meningococcal disease (145.9, 95% CI 116.7–178.3); legionellosis (43.3, 95% CI 28.0–62.0); and COVID-19 (21.9, 95% CI 19. 7–24.3). Most diseases exhibited a strong negative gradient, with high SMRs in the first 7-days of illness onset that reduced over time. Interpretation: We found evidence of substantially elevated all-cause mortality among people notified with communicable diseases. Not all elevated risk is likely directly attributable to the communicable diseases of interest, rather, it may reflect underlying comorbidities or behaviours in these individuals. Regardless of attribution, infection with communicable diseases may represent a marker of mortality. Key to preventing deaths may be through timely and appropriate transition to primary and preventive healthcare following diagnosis. Funding: No funding was provided for this study. Declaration of Interest: None to declare. Ethical Approval: Study approvals were obtained from the Victorian Government Department of Health Research Ethics Committee (reference: LNR/47982), and registered with Monash University Human Research Ethics Committee (reference: 30076)·

ABSTRACT BACKGROUND Paediatric neuroblastoma and brain tumours account for a third of all childhood cancer-related mortality. High-risk neuroblastoma is highly aggressive and survival is poor despite intensive multi-modal therapies with significant toxicity. Novel therapies are desperately needed. The Zika virus (ZIKV) is neurotropic and there is growing interest in employing ZIKV as a potential therapy against paediatric nervous system tumours, including neuroblastoma. METHODS Here, we perform an extensive meta-analysis of ZIKV infection studies to identify molecular mechanisms that may govern the oncolytic response in neuroblastoma cells. We summarise the neuroblastoma cell lines and ZIKV strains utilised and re-evaluate the infection data to deduce the susceptibility of neuroblastoma to the ZIKV oncolytic response. Integrating transcriptomics, interaction proteomics, dependency factor and compound datasets we show the involvement of multiple host systems during ZIKV infection. RESULTS We identify that most paediatric neuroblastoma cell lines are highly susceptible to ZIKV infection and that the PRVABC59 ZIKV strain is the most promising candidate for neuroblastoma oncolytic virotherapy. ZIKV induces TNF signalling, lipid metabolism, the Unfolded Protein Response (UPR), and downregulates cell cycle and DNA replication processes. ZIKV is dependent on SREBP-regulated lipid metabolism and three protein complexes; V-ATPase, ER Membrane Protein Complex (EMC) and mammalian translocon. We propose ZIKV nonstructural protein 4B (NS4B) as a likely mediator of ZIKVs interaction with IRE1-mediated UPR, lipid metabolism and mammalian translocon. CONCLUSIONS Our work provides a significant understanding of ZIKV infection in neuroblastoma cells, which will facilitate the progression of ZIKV-based oncolytic virotherapy through pre-clinical research and clinical trials. KEYPOINTS The Zika virus may provide the basis for an oncolytic virotherapy against Neuroblastoma Most paediatric neuroblastoma cell lines are susceptible to Zika viral infection We identified molecular mechanisms that may induce the oncolytic response in Neuroblastoma Contribution to the field The ability to both induce direct oncolysis and provoke an anti-tumoral immune response makes oncolytic virotherapy an attractive candidate to combat aggressive and heterogenous cancers, such as high-risk neuroblastoma. To progress oncolytic virotherapy to clinical trial it is essential to understand the host mechanisms the virus manipulates to kill cancer cells, alongside any pathology as a consequence of infection of normal cells. Here, we show that ZIKV efficiently infects and induces oncolysis of paediatric neuroblastoma cells and propose a potential TNF pathway-driven immune response. ZIKV’s specificity for infection of nervous system cancer cells, while rarely causing nervous system-related pathology in young children, addresses many of its safety concerns. The inclusion of more effective and less toxic novel therapies, such as a potential ZIKV-based therapeutic, in multimodal treatment regimens will pave the way for improving patient long-term health and overall survival.

ABSTRACT Viruses are known to co-opt host machinery for translation initiation, but less is known about which host factors are required for the formation of ribosomes used to synthesize viral proteins. Using a loss of function CRISPR screen, we show that synthesis of a flavivirus encoded reporter depends on multiple host factors, including several 60S ribosome biogenesis proteins. Viral phenotyping revealed that two of these factors, SBDS, a known ribosome biogenesis factor, and the relatively uncharacterized protein SPATA5, were broadly required for replication of flaviviruses, coronaviruses, alphaviruses, paramyxoviruses, an enterovirus, and a poxvirus. Mechanistic studies revealed that loss of SPATA5 caused defects in ribosomal RNA processing and ribosome assembly, suggesting that this human protein may be a functional ortholog of yeast Drg1 . These studies implicate specific ribosome biogenesis proteins as viral host dependency factors that are required for synthesis of virally encoded protein and accordingly, optimal viral replication.

ABSTRACT Introduction Smoking is one of the lifestyle choices associated with an increased risk of chronic health conditions and poorer COVID-19 outcomes. Because it is known that the lungs recover after quitting smoking, a direct comparison of the severity of COVID-19 infection in current and former smokers needs to be investigated. Methods and analysis The Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocol (PRISMA-P) 2015 Checklist was used. Non-randomized studies will be searched in PubMed, Cochrane CENTRAL library, Embase, and Epistemonikos from December 2019 to the present. Hand-searching of grey literature, key journals, and reference lists will be conducted This review will include studies of current and former smokers, with the main outcome being ICU admission, assisted respiration, or death. Two independent reviewers will select primary studies and abstract data from them. The Newcastle-Ottawa checklist will be used to assess the risk of bias, and the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) framework will be used to assess the quality of cumulative evidence. RevMan 5.4 will be used for data analysis. The I 2 statistic will be used to evaluate heterogeneity. For similar studies, the fixed-effect method of a meta-analysis will be used; otherwise, a random-effect model will be used. The qualitative synthesis will be used for studies that are ineligible for the quantitative approach. Ethical consideration and dissemination Because published data will be reviewed, no ethical approval is required. Our findings will be presented at national and/or international conferences, and they will be published in a peer-reviewed journal. PROSPERO registration number CRD42022368552 ARTICLE SUMMARY Strengths and limitations of this study This is a focused research question comparing the current and ex-smokers risk of contracting the severe form of COVID-19. This systematic review and meta-analysis will provide evidence of the dangers of smoking during the COVID-19 pandemic. The PRISMA-P reporting guidelines were strictly followed while writing this protocol. Study selection will be carried out by two independent reviewers and a third person will intervene if a disagreement arises. A potential limitation is that an observational study design will be used in this systematic review.

Social media has been one of the main information consumption sources for the public, allowing people to seek and spread information more quickly and easily. However, the rise of various social media platforms also enables the proliferation of online misinformation. In particular, misinformation in the health domain has significant impacts on our society such as the COVID-19 infodemic. Therefore, health misinformation in social media has become an emerging research direction that attracts increasing attention from researchers of different disciplines. Compared to misinformation in other domains, the key differences of health misinformation include the potential of causing actual harm to humans' bodies and even lives, the hardness to identify for normal people, and the deep connection with medical science. In addition, health misinformation on social media has distinct characteristics from conventional channels such as television on multiple dimensions including the generation, dissemination, and consumption paradigms. Because of the uniqueness and importance of combating health misinformation in social media, we conduct this survey to further facilitate interdisciplinary research on this problem. In this survey, we present a comprehensive review of existing research about online health misinformation in different disciplines. Furthermore, we also systematically organize the related literature from three perspectives: characterization, detection, and intervention. Lastly, we conduct a deep discussion on the pressing open issues of combating health misinformation in social media and provide future directions for multidisciplinary researchers.

The worldwide expansion of mosquito-borne pathogens necessitates improved control measures, including approaches to reduce transmission by mosquito vectors. Reducing transmission is challenging because determinants of vector competence for viruses like Zika (ZIKV) are poorly understood. Our previous work established that Aedes (Ae.) aegypti larvae reared in environmental water containing microbes are less susceptible to ZIKV as adults compared to cohorts reared in laboratory tap water with fewer microbial species and lower microbial abundance. Here, we identify a process by which environment-derived microbes reduce susceptibility of Ae. aegypti for ZIKV. Provided that the midgut represents the first barrier to mosquito infection, we hypothesized that microbial exposure modulates midgut infection by ZIKV. Since mosquitoes live in water as larvae and pupae and then transition to air as adults, we also define the stage in the life of a mosquito when microbial exposure reduces ZIKV susceptibility. Ae. aegypti larvae were reared in water containing microbes and then treated with antibiotics during the pupal and adult stages, adult stage only, or provided no antibiotics at any stage. Vector competence was next evaluated in mosquitoes that ingested ZIKV-spiked bloodmeals. Antibiotic treated mosquitoes with reduced microbiota showed enhanced ZIKV infection rates in Ae. aegypti treated as both pupae and adults. Antibiotic treatment to disrupt microbes in pupal and adult mosquitoes also resulted in increased midgut epithelium permeability, higher numbers of ZIKV-infected midgut cells, and impaired bloodmeal digestion. Parallel control experiments with antibiotic-treated or gnotobiotic mosquitoes reared in laboratory water showed that the dysbiotic state created by antibiotic use does not influence ZIKV vector competence or midgut permeability and that more than the bacterial species in gnotobiotic mosquitoes is responsible for reducing ZIKV vector competence. Ae. aegypti with disrupted microbiota via antibiotic treatment as pupae and adults that ingested ZIKV in bloodmeals showed reduced expression of genes associated with bloodmeal digestion and metabolism relative to mosquitoes whose microbes were not reduced with antibiotics. Together, these data show that exposure to microbes throughout the life of Ae. aegypti restricts ZIKV infection by facilitating blood digestion and reducing midgut cell infection. Understanding the connections between mosquito microbiota, midgut physiology, and arbovirus susceptibility can lead to novel approaches to decrease mosquito transmission and will improve understanding of vector competence in environmental habitats containing microbes. Author Summary Mosquito-transmitted viruses like Zika continue to threaten human health. Absent vaccines or treatments, controlling mosquitoes or limiting their ability to transmit viruses represents a primary way to prevent mosquito-borne viral diseases. The role mosquito microbiota play in shaping transmission of Zika virus has been limited to association-based studies. Our prior work showed that Aedes aegypti mosquito larvae that develop in water containing bacteria are less susceptible to Zika virus compared to larvae reared in laboratory tap water with fewer numbers and species of bacteria. Here we identify a process that explains this association. Since mosquitoes are aquatic as larvae and pupae and terrestrial as adults, we also define the life stage when microbes need be present to reduce Zika virus susceptibility. We used antibiotics to reduce environmental water-derived microbes at pupal and adult or only adult stages and observed that microbial disruption via antibiotic treatment increases Zika virus infection and midgut permeability and impairs bloodmeal digestion. These findings advance understanding of microbiota-mosquito-virus interactions and further implicate microbes as a means to restrict virus infection of mosquitoes.

Zika virus (ZIKV) infection causes severe neurological consequences in both gestationally-exposed infants and adults. Sensorial gating deficits strongly correlate to the motor, sensorial and cognitive impairments observed in ZIKV-infected patients. However, to date, no startle response or prepulse inhibition (PPI) assessment has been made in patients or animal models. In this study, we identified different outcomes for the age of infection and sex in wild-type mice: neonatally infected animals presented an increase in PPI and startle latency in both sexes, while adult infected males presented lower startle amplitude but preserved PPI. Our data further the understanding of the functional impacts of ZIKV on the developing and mature nervous system, which could help explain other behavioral and cognitive alterations caused by the virus. With this study, we support the use of startle reflex testing in ZIKV exposed patients, especially infants, allowing for early detection of functional neuromotor damage and early intervention.

This chapter examines the evolving implementation of the International Health Regulations (IHR) in responding to public health emergencies. Analyzing implementation obligations under global health law, Part II outlines the national and global governance expectations inherent in the IHR. Part III chronicles the evolving implementation of the IHR in responding to infectious disease threats since the establishment of WHO, with early efforts seen as largely ineffective in speaking to the emerging disease threats of a globalizing world. Drawing from the sweeping changes in implementation expectations promulgated by IHR (2005), Part IV examines the implications of the revised IHR in responding to a rapid succession of public health emergencies of international concern. Yet despite continuing advancements in IHR (2005) implementation over fifteen years, the COVID-19 pandemic overwhelmed the world, and Part V examines the limitations of the IHR in responding to this cataclysmic threat, raising an imperative for further reforms of global health law. As the world considers IHR revisions anew, with these revisions taking place amid an interconnected series of multisectoral reforms in global health law and governance, this chapter concludes that past limitations point the way forward in revising institutional mechanisms, ensuring IHR implementation as a foundation for global health security under global health law.

Abstract Zika virus (ZIKV) infections have been associated with severe clinical outcomes, including neurological manifestations, especially in newborns with intrauterine infection. However, no licensed vaccines or specific antiviral agents are available yet. Therefore, safe and low-cost therapy is required, especially for pregnant women. In this sense, metformin, an FDA-approved drug used to treat gestational diabetes, has previously exhibited an effect anti-ZIKA in vitro in HUVEC cells by activating AMPK. In this study, we evaluated metformin treatment during in vitro ZIKV infection in a permissive trophoblast cell line JEG3. Our results demonstrate that metformin impacts viral replication and protein synthesis and reverts the cytoskeletal changes promoted by ZIKV infection. Beyond this, lipid droplet formation is reduced, which is associated with the lipogenic activation of infection. Taken together, our findings indicate that metformin has potential as an antiviral agent against in vitro ZIKV infection in trophoblastic cells.

Background: Vector-borne flaviviruses, including tick-borne encephalitis virus (TBEV), Zika virus (ZIKA), West Nile virus (WNV), yellow fever virus (YFV), dengue virus (DENV), and Japanese encephalitis virus (JEV), pose a growing threat to public health worldwide, and have evolved complex mechanisms to overcome host antiviral innate immunity. However, the underlying mechanisms of flavivirus structural proteins to evade host immune response remain elusive. Results: : We show that TBEV structural protein, pre-membrane (prM) protein, could inhibit type I interferon (IFN-I) production. Mechanically, TBEV prM interacted with both MDA5 and MAVS and interfered with the formation of MDA5-MAVS complex, thereby impeding the nuclear translocation and dimerization of IRF3 to inhibit RLR antiviral signaling. ZIKA and WNV prM was also demonstrated to interact with both MDA5 and MAVS, while dengue virus serotype 2 (DENV2) and YFV prM associated only with MDA5 or MAVS to suppress IFN-I production. In contrast, JEV prM could not suppress IFN-I production. Overexpression of TBEV and ZIKA prM significantly promoted the replication of TBEV and Sendai virus. Conclusion: Our findings reveal the immune evasion mechanisms of flavivirus prM, which may contribute to understanding flavivirus pathogenicity, therapeutic intervention and vaccine development.

Background: Zika Virus (ZIKV), an arbovirus of global concern, has been associated with neurological complications including microcephaly in newborns and Guillain-Barré Syndrome in adults. Like other flaviviruses, ZIKV depends on cholesterol to facilitate its replication; thus, cholesterol has been proposed as a therapeutic target to treat the infection using FDA-approved statins. Cholesterol is stored in intracellular lipid droplets (LD) in the form of cholesterol esters and can be regulated by autophagy. We hypothesize that the virus hijacks autophagy machinery as an early step to increase the formation of LD and viral replication, and that interference with this pathway will limit reproduction of virus. Methods: : We pretreated MDCK cells with atorvastatin or other inhibitors of autophagy prior to infection with ZIKV. We measured viral expression by qPCR for NS1 RNA and immunofluorescence for Zika E protein. Results: : Autophagy increases in virus-infected cells as early as 6 hours post infection (hpi). In the presence of atorvastatin, LD are decreased, and cholesterol is reduced, targeting key steps in viral replication, resulting in suppression of replication of ZIKV is suppressed. Other both early- and late-acting autophagy inhibitors decrease both the number of LD and viral replication. Bafilomycin renders cholesterol is inaccessible to ZIKV. We also confirm previous reports of a bystander effect, in which neighboring uninfected cells have higher LD counts compared to infected cells. Conclusions: : We conclude that atorvastatin and inhibitors of autophagy lead to lower availability of LD, decreasing viral replication. We conclude that bafilomycin A1 inhibits viral expression by blocking cholesterol esterification to form LD.

Background: Post-infectious autoimmunity is a hallmark of Guillain–Barré syndrome (GBS), and GBS incidence closely parallels that of its immune triggers. Sociobehavioral interventions implemented during the coronavirus disease pandemic have altered the infectious disease landscape. Methods: This nationwide longitudinal, time-series correlation study analyzed GBS incidence and sentinel surveillance data from January 2017 to December 2021 in the National Health Insurance Service and Korean Disease Control and Prevention Agency databases. The incidence of GBS and sentinel gastrointestinal and respiratory infectious diseases during the pandemic (2020–2021) was estimated and compared with both pre-pandemic (2017–2019) and incidence predicted in a time-series forecasting model to examine temporal associations between GBS and infectious triggers. Findings: During the pandemic, the total crude cumulative incidence rate (CIR) was 2.1 per 100,000 population, which is lower than the pre-pandemic incidence, especially in age groups of less than 60 years. Seasonality was briefly interrupted during the winter of 2021. The majority of common infectious diseases, and respiratory more than gastrointestinal conditions, had a lower-than-expected incidence during the pandemic. Compared to the pre-pandemic state, during the pandemic period a higher number of gastrointestinal pathogens ( E. coli, Campylobacter spp., C. perfringens, Y. enterocolitica , and enteric adenovirus) had significant, moderate-to-strong positive temporal associations with GBS. Interpretation: The incidence of GBS and sentinel infectious diseases decreased to below-expected levels during the pandemic, with the former attributable to the decreased incidence of non-COVID-19 respiratory and gastrointestinal infections. The evolving incidence of autoimmune post-infectious phenomena following the pandemic needs attention. Funding: Korea Disease Control and Prevention Agency Declaration of Interest: The authors declare no conflict of interests. Ethical Approval: The Institutional Review Board at KDCA exempted this study from ethical approval because of the retrospective analysis of de-identified data that were already obtained through epidemiological investigation, presented minimal privacy or confidentiality risk to participants, and met the current public health interest requirements. All study procedures have been reported in accordance with the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Reporting Guidelines. The requirement for consent was waived because we retrospectively collected de-identified data.

This research focused on the Indonesian government’s global pandemic policies mapping from 2000-2021. Indonesia has been affected by some diseases such as H1N1, H5N1, SARS-Cov-1, and the current SARS- CoV-2 (Covid-19). These three outbreaks will be the main focus of this research in order to see the readiness of Indonesian government policy mapping to emergencies. This research will measure Indonesian health policy mapping based on its capability to adapt to the local context, construct a care delivery value chain, leverage shared delivery infrastructure, and improve health delivery and economic development. We perform feasibility analysis with a method scoring system to the fourth Pillar above. The result of this research indicated that most of the health policies taken by the Indonesian government are oriented on the response to a current condition, not preparing for future health challenges. At the end of this paper, this research will also provide patterns and descriptions regarding the development of health policy-making in order to deal with emergency conditions caused by pandemics or certain viral epidemics. Keywords: health policy, policy analysis, disease control, health care, emergency service, effectiveness, emergent infections, risk situation, policy implementation

Advances in AI technologies have resulted in superior levels of AI-based model performance. However, this has also led to a greater degree of model complexity, resulting in 'black box' models. In response to the AI black box problem, the field of explainable AI (xAI) has emerged with the aim of providing explanations catered to human understanding, trust, and transparency. Yet, we still have a limited understanding of how xAI addresses the need for explainable AI in the context of healthcare. Our research explores the differing explanation needs amongst stakeholders during the development of an AI-system for classifying COVID-19 patients for the ICU. We demonstrate that there is a constellation of stakeholders who have different explanation needs, not just the 'user'. Further, the findings demonstrate how the need for xAI emerges through concerns associated with specific stakeholder groups i.e., the development team, subject matter experts, decision makers, and the audience. Our findings contribute to the expansion of xAI by highlighting that different stakeholders have different explanation needs. From a practical perspective, the study provides insights on how AI systems can be adjusted to support different stakeholders needs, ensuring better implementation and operation in a healthcare context.