ABSTRACT SARS-CoV-2, responsible for the COVID-19 pandemic, causes respiratory failure and damage to multiple organ systems. The emergence of viral variants poses a risk of vaccine failures and prolongation of the pandemic. However, our understanding of the molecular basis of SARS-CoV-2 infection and subsequent COVID-19 pathophysiology is limited. In this study, we have uncovered a critical role for the evolutionarily conserved Hippo signaling pathway in COVID-19 pathogenesis. Given the complexity of COVID-19 associated cell injury and immunopathogenesis processes, we investigated Hippo pathway dynamics in SARS-CoV-2 infection by utilizing COVID-19 lung samples, and human cell models based on pluripotent stem cell-derived cardiomyocytes (PSC-CMs) and human primary lung air-liquid interface (ALI) cultures. SARS-CoV-2 infection caused activation of the Hippo signaling pathway in COVID-19 lung and in vitro cultures. Both parental and Delta variant of concern (VOC) strains induced Hippo pathway. The chemical inhibition and gene knockdown of upstream kinases MST1/2 and LATS1 resulted in significantly enhanced SARS-CoV-2 replication, indicating antiviral roles. Verteporfin a pharmacological inhibitor of the Hippo pathway downstream transactivator, YAP, significantly reduced virus replication. These results delineate a direct antiviral role for Hippo signaling in SARS-CoV-2 infection and the potential for this pathway to be pharmacologically targeted to treat COVID-19.

The systemic nature of SARS-CoV-2 infection is highly recognized, but poorly characterized. A non-invasive and unbiased method is needed to clarify whole body spatiotemporal dynamics of SARS-CoV-2 infection after transmission. We recently developed a probe based on the anti-SARS-CoV-2 spike antibody CR3022 to study SARS-CoV-2 pathogenesis in vivo. Herein, we describe its use in immunoPET to investigate SARS-CoV-2 infection of three rhesus macaques. Using PET/CT imaging of macaques at different times post-SARS-CoV-2 inoculation, we track the 64Cu-labelled CR3022-F(ab’)2 probe targeting the spike protein of SARS-CoV-2 to study the dynamics of infection within the respiratory tract and uncover novel sites of infection. Using this method, we uncovered differences in lung pathology between infection with the WA1 isolate and the delta variant, which were readily corroborated through computed tomography scans. The 64Cu-CR3022-probe also demonstrated dynamic changes occurring between 1- and 2-weeks post-infection. Remarkably, a robust signal was seen in the male genital tract (MGT) of all three animals studied. Infection of the MGT was validated by immunofluorescence imaging of infected cells in the testicular and penile tissue and severe pathology was observed in the testes of one animal at 2-weeks post-infection. The results presented here underscore the utility of using immunoPET to study the dynamics of SARS-CoV-2 infection to understand its pathogenicity and discover new anatomical sites of viral replication. We provide direct evidence for SARS-CoV-2 infection of the MGT in rhesus macaques revealing the possible pathologic outcomes of viral replication at these sites.

Recent infectious disease outbreaks, such as the COVID-19 pandemic and the Zika epidemic in Brazil, have demonstrated both the importance and difficulty of accurately forecasting novel infectious diseases. When new diseases first emerge, we have little knowledge of the transmission process, the level and duration of immunity to reinfection, or other parameters required to build realistic epidemiological models. Time series forecasts and machine learning, while less reliant on assumptions about the disease, require large amounts of data that are also not available in early stages of an outbreak. In this study, we examine how knowledge of related diseases can help make predictions of new diseases in data-scarce environments using transfer learning. We implement both an empirical and a theoretical approach. Using empirical data from Brazil, we compare how well different machine learning models transfer knowledge between two different disease pairs: (i) dengue and Zika, and (ii) influenza and COVID-19. In the theoretical analysis, we generate data using different transmission and recovery rates with an SIR compartmental model, and then compare the effectiveness of different transfer learning methods. We find that transfer learning offers the potential to improve predictions, even beyond a model based on data from the target disease, though the appropriate source disease must be chosen carefully. While imperfect, these models offer an additional input for decision makers during pandemic response.

Background and Rationale Approximately 35% of all COVID-19 deaths occurred in Skilled Nursing Facilities (SNFs). In a healthy general population, wearables have shown promise in providing early alerts for actionable interventions during the pandemic. We tested this promise in a cohort of SNFs patients diagnosed with COVID-19 and admitted for post-acute care under quarantine. We tested if 1) deployment of wearables and contact-free biosensors is feasible in the setting of SNFs and 2) they can provide early and actionable insights into deterioration. Methods This prospective clinical trial has been IRB-approved ( NCT04548895 ). We deployed two commercially available devices detecting continuously every 2-3 minutes heart rate (HR), respiratory rate (RR) and uniquely providing the following biometrics: 1) the wrist-worn bracelet by Biostrap yielded continuous oxygen saturation (O2Sat), 2) the under-mattress ballistocardiography sensor by Emfit tracked in-bed activity, tossing, and sleep disturbances. Patients also underwent routine monitoring by staff every 2-4 h. For death outcomes, cases are reported due to the small sample size. For palliative care versus at-home discharges, we report mean±SD at p<0.05. Results From 12/2020 - 03/2021, we approached 26 PCR-confirmed SarsCoV2-positive patients at two SNFs: 5 declined, 21 were enrolled into monitoring by both sensors (female=13, male=8; age 77.2±9.1). We recorded outcomes as discharged to home (8, 38%), palliative care (9, 43%) or death (4, 19%). The O2Sat threshold of 91% alerted for intervention. Biostrap captured hypoxic events below 91% nine times as often as the routine intermittent pulse oximetry. In the patient deceased, two weeks prior we observed a wide range of O2Sat values (65-95%) captured by the Biostrap device and not noticeable with the routine vital sign spot checks. In this patient, the Emfit sensor yielded a markedly reduced RR (7/min) in contrast to 18/min from two routine spot checks performed in the same period of observation as well as compared to the seven patients discharged home over a total of 86 days of monitoring (RR 19± 4). Among the patients discharged to palliative care, a total of 76 days were monitored, HR did not differ compared to the patients discharged home (68±8 vs 70±7 bpm). However, we observed a statistically significant reduction of RR at 16±4/min as well as the variances in RR (10±6 vs 19±4/min vs16±13) and activity of palliative care patients vs. patients discharged home. Conclusion/Discussion We demonstrate that wearables and under-mattress sensors can be integrated successfully into the SNF workflows and are well tolerated by the patients. Moreover, specific early changes of oxygen saturation fluctuations and other biometrics herald deterioration from COVID-19 two weeks in advance and evaded detection without the devices. Wearable devices and under-mattress sensors in SNFs hold significant potential for early disease detection.

Flaviviruses replicate in membrane factories associated with the endoplasmic reticulum (ER). Significant levels of flavivirus viral protein accumulation contribute to ER stress. As a consequence, the host cell exhibits an Unfolded Protein Response (UPR), subsequently stimulating appropriate cellular responses such as adaptation, autophagy or apoptosis. The correct redox conditions of this compartment are essential to form native disulfide bonds in proteins. ZIKA virus (ZIKV) has ability to induce persistent ER stress leading to activation of UPR pathways. In this study, we wondered whether ZIKV affects the redox balance and consequently the oxidative protein folding in the ER. We found that ZIKV replication influences redox state leading to aggregation of viral envelope protein as amyloid-like structures in the infected cells.

Background Understanding the underlying risk of infection by dengue virus from surveillance systems is complicated due to the complex nature of the disease. In particular, the probability of becoming severely sick is driven by serotype-specific infection histories as well as age; however, this has rarely been quantified. Island communities that have periodic outbreaks dominated by single serotypes provide an opportunity to disentangle the competing role of serotype, age and changes in surveillance systems in characterising disease risk. Methodology We develop mathematical models to analyse 35 years of dengue surveillance (1979-2014) and seroprevalence studies from French Polynesia. We estimate the annual force of infection, serotype-specific reporting probabilities and changes in surveillance capabilities using the annual age and serotype-specific distribution of dengue. Principal Findings Eight dengue epidemics occurred between 1979 and 2014, with reporting probabilities for DENV-1 primary infections increasing from 3% to 5%. The reporting probability for DENV-1 secondary infections was 3.6 times that for primary infections. Reporting probabilities for DENV-2–DENV-4 were 0.1-2.6 and 0.7-2.3 times that for DENV-1, for primary and secondary infections, respectively. Reporting probabilities declined with age after 14 y.o. Between 1979 and 2014, the proportion never infected declined from 70% to 23% while the proportion infected at least twice increased from 4.5% to 45%. By 2014, almost half of the population had acquired heterotypic immunity. The probability of an epidemic increased sharply with the estimated fraction of susceptibles among children. Conclusion / Significance By analysing 35 years of dengue data in French Polynesia, we characterised key factors affecting the dissemination profile and reporting of dengue cases in an epidemiological context simplified by mono-serotypic circulation. Our analysis provides key estimates that can inform the study of dengue in more complex settings where the co-circulation of multiple serotypes can greatly complicate inference. Author summary Characterising the true extent of dengue circulation and the level of population immunity is essential to assess the burden of disease, evaluate epidemic risk and organise prevention strategies against future epidemics. However, this is difficult in a context where most people who are infected by dengue virus (DENV) only have mild symptoms which may not be reported to surveillance systems. In this article, we develop a mathematical model to evaluate the fraction of unreported dengue infections from case data. The key idea is to introduce reporting probabilities that depend on the infecting serotype and the infection history of patients. These factors are known to contribute to variations in the severity of symptoms and hence the reporting probabilities, but have rarely been taken into account in model frameworks to study population immunity from the case data. Using the developed model, we study long-term dengue virus immunity in French Polynesia.

Zika virus (ZIKV) infection is a public health concern and currently there is no specific therapeutic or approved vaccine. Resveratrol (RESV), a natural antiviral compound, has been shown to possess antiviral properties, but the mechanisms of action against ZIKV remain unknown. This study aimed to investigate the role of the high mobility group box 1 protein (HMGB1) in the underlying anti-ZIKV mechanisms of RESV. HMGB1 protein expression and ZIKV replication in both RESV-treated wild-type (WT) and HMGB1-knockdown (shHMGB1) Huh7 cells were analyzed using ELISA, immunofluorescence assay, immunoblot assay, focus-forming assay and qRT-PCR. HMGB1’s role was explored by evaluating the changes in the type-1 interferon (IFN) response genes using qRT-PCR and immunoblot assays. The treatment of ZIKV-infected WT Huh7 cells with RESV significantly reduced ZIKV titers by >90% (P < 0.001) and inhibited ZIKV-induced HMGB1 translocation (P < 0.001), resulting in nuclear HMGB1 accumulation. Compared to WT Huh7 cells, shHMGB1 Huh7 cells without RESV treatment showed a significant increase in the infectious virus titers and RNA with a maximum rise of 74% (P < 0.001) and 65% (P < 0.01), respectively. RESV treatment of ZIKV-infected WT Huh7 cells significantly increased the MxA (one of the classical interferon-stimulated genes, ISGs) and IFN-β levels (p < 0.05). The treatment of infected shHMGB1 Huh7 cells with RESV showed a less effective antiviral response (P > 0.05) and did not cause changes in the expressions of MxA and IFN-β. Findings presented here suggest that RESV possesses therapeutic activity against ZIKV infection and the mechanism of action is mainly attributed to HMGB1 nuclear retention, which could upregulate the type-1 IFN and ISGs.

Background: In 2018, the USAID Applying Science to Strengthen and Improve Systems (ASSIST) Project started a new partnership with four Eastern and Southern Caribbean countries impacted by the Zika virus: Antigua and Barbuda, Dominica, St. Kitts and Nevis, and St. Vincent and the Grenadines. The goal of the project was to provide short-term technical assistance (STTA) to strengthen the health systems’ capacity to detect newborns and young children potentially affected by Zika and to address their health needs. To meet these objectives, ASSIST developed an innovative approach based on its existing model for service delivery improvement. This novel approach is known as Rapid, Multi-country, Parallel Process, Multi-tasking Approach for a Project Startup (RMPP-MAPS). An evaluation was conducted to document the STTA startup activities, to identify enabling and constraining factors, and to capture lessons learned. Methods: : An external consultant conducted remote in-depth interviews with individuals involved in the startup using semi-structured interview guides and retrieved data from the review of project documents. Results: : Using RMPP-MAPS, the ASSIST Project successfully implemented the startup for complex STTA in four countries within less than four months, spanning mid-May to early September 2018. Project milestones included achieving buy-in from stakeholders, co-developing the technical scope and materials, and rapidly executing critical operational functions. Dedicated project teams, country leaderships, and local champions were essential to overcoming the main challenges, which included a condensed timeframe, lack of in-country offices, and country-level factors such as a shortage of health care workers and a weak health infrastructure. Conclusions: The RMPP-MAPS is a feasible and resource-efficient mechanism of interest to implementers, donors, and low and middle-income countries facing temporal and financial limitations to rapidly addressing public health priorities.

Flavivirus infections show recurrent outbreaks and can be responsible for disease complications such as Hemorrhagic Dengue Fever and Congenital Zika Virus Syndrome. Effective therapeutic interventions are still a challenge. Antibodies can provide significant protection, although antibody response may fail due to ADE (Antibody-Dependent Enhancement) reactions or immune escape mutations. To generate effective neutralizing antibodies, the choice of the target antigen is a crucial part of the process. Human anti-Flavivirus antibodies were selected from a combinatorial library displayed on a phage surface. The antibodies were selected against a mimetic peptide based on the fusion loop region in Domain II of the protein E, which is highly conserved among different Flavivirus. Four rounds of selection were performed using the synthetic peptide in two strategies: the first was using acidic elution of bound phages, and the second was applying a competing procedure. After panning, the selected VH and VL domains were determined by combining NGS and bioinformatic approaches. Three different human monoclonal antibodies were expressed as scFvs and further characterized. All showed binding capacity to Zika (ZIKV), Yellow Fever (YFV), and Dengue (DENV) viruses. Two of these antibodies, AZ1p and AZ6m, could neutralize the ZIKV infection in a PRNT assay. These new antibodies have the potential to be used in therapeutic intervention against different Flavivirus illnesses and, due to the conservation of the fusion loop region, they may be resistant to scape mutations. Author summary The central idea of this work is to present a possible unique therapeutic approach to combat different diseases that cause health problems annually, such as Dengue and yellow fever infections and Zika congenital syndromes. The viruses that cause these diseases, of the Flavivirus genus, typically have disease amplification reactions and evasion mechanisms of the immune response that hinder the success of specific therapies and vaccines and require new forms of effective and safe treatments. We have developed new neutralizing human antibodies so that they bind to a highly conserved region of Flavivirus, called the fusion loop, and in such a way as to avoid adverse effects associated with anti-Flavivirus antibodies. We showed that the antibodies have high cross-reactivity against different Flaviviruses and we exemplified their neutralizing capacity for Zika virus infection in an ex vivo assay. New monoclonal antibodies such as those presented here may contribute to the control of important tropical diseases in a safer and more efficient way.

Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has caused many complications, the invention of coronavirus disease 2019 (Covid-19) vaccines has also brought about several adverse events, from common side effects to unexpected and rare ones. Common vaccine-related adverse reactions manifest locally or systematically following any vaccine, including Covid-19 vaccines. Certain side effects, known as adverse events of special interest (AESI), are unusual and need more evaluation. Here, we discuss some of the most important rare adverse events of Covid-19 vaccines.

The COVID-19 pandemic has highlighted an urgent need for new antivirals. Targeting host processes co-opted by viruses is an attractive antiviral strategy with a high resistance barrier. Picolinic acid (PA) is a Tryptophan metabolite endogenously produced in mammals. Here we report broad-spectrum antiviral activity of PA against enveloped viruses including Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), Influenza A virus (IAV), Flaviviruses, Herpes simplex virus, and Parainfluenza virus. Mechanistic studies revealed that PA inhibits enveloped virus entry by compromising viral membrane integrity, inhibiting virus-cellular membrane fusion, and interfering with cellular endocytosis. More importantly, in preclinical animal models, PA showed promising antiviral efficacy against SARS-CoV-2 and IAV, especially as an oral prophylactic. Overall, our data establish PA as a broad-spectrum antiviral, with promising preclinical efficacy against pandemic viruses SARS-CoV-2 and IAV. Funding Information: DBT-Wellcome Trust India Alliance (IA/I/18/1/503613), DBT-BIRAC (BT/CS0007/CS/02/20), Crypto Relief Fund, L & T Trust, DST-FIST program, DBT-IISc partnership program (Phase II), Bacteriophage work in the RA laboratory was supported by Ramanujan Fellowship (SB/S2/RJN-037/2017). Conflict of Interests: Indian Institute of Science has filed patents for use of Picolinic acid and its derivatives as broad-spectrum antiviral and listed ST, RN, AB, and RR as inventors. Ethical Approval: This study was conducted in compliance with institutional biosafety guidelines, (IBSC/IISc/ST/17/2020; IBSC/IISc/ST/18/2021), following the Indian Council of Medical Research and Department of Biotechnology recommendations. All experiments involving animals were reviewed and approved by the Institutional Animal Ethics Committee (Ref: IAEC/IISc/ST/784/2020) at the Indian Institute of Science and conducted in Viral Biosafety level-3 facility. The experiments were performed according to CPCSEA (The Committee for Control and Supervision of Experiments on Animals) guidelines.

In regard to the global pandemic of COVID-19, it seems that persons with epilepsy (PWE) are not more vulnerable to get infected by SARS-CoV-2, nor are they more susceptible to a critical course of the disease. However, management of acute seizures in patients with COVID-19 as well as management of PWE and COVID-19 needs to consider potential drug-drug interactions between antiseizure drugs and candidate drugs currently assessed as therapeutic options for COVID-19. Repurposing of several licensed and investigational drugs is discussed for therapeutic management of COVID-19. While for none of these approaches, efficacy and tolerability has been confirmed yet in sufficiently powered and controlled clinical studies, testing is ongoing with multiple clinical trials worldwide. Here, we have summarized the possible mechanisms of action of drugs currently considered as potential therapeutic options for COVID-19 management along with possible and confirmed drug-drug interactions that should be considered for a combination of antiseizure drugs and COVID-19 candidate drugs.

Although many therapies have been proposed, there is no evidence of any effective treatment for COVID-19 to date. Currently, the main therapies being used to treat the critically patients with COVID-19 are antiviral drugs, chloroquine/hydroxychloroquine and respiratory therapy. Pharmacokinetics (PK) and pharmacodynamics (PD) play an important role in different clinical situations, and are keys to balance the effect (reduction of SARS-CoV-2 virus and symptom improvement) and toxicity (adverse effects). Critically ill patients always have altered PK and PD due to multiple factors such as hypoproteinemia, organ dysfunction, and organ support treatment including continuous renal replacement therapy (CRRT), and extracorporeal membrane oxygenation (ECMO). Here we provide an overview of the effects of the clinical treatments for critically COVID-19 patients and the pathological state of patients on pharmacokinetics and pharmacodynamics parameters of therapeutic drugs in detail. However, current evidence is insufficient to speculate the changes of drug concentration of antimicrobials, therapeutic drug monitoring (TDM) is a useful tool used in severe patients since many suspected factors were exist to influence the clinical responses and adverse drug reactions of drugs.

After the outbreak of the Coronavirus, because of travel restriction and fear, it can be anticipated that there would be a significant change in travel behaviour. However, very few research exclusively analyzed the changes of factors for different modes at different stages of COVID-19 (before, during, and after) in the context of developing countries. This study contributed to the knowledge, first by understanding if any changes in travel patterns transpired among the stages of the outbreak (before, during, and after), and second by understanding the importance of variables for different travel modes during each stage for different cities in Bangladesh. The research questions that were answered to achieve this aim are 1) Do travel patterns vary before, during, and after the COVID-19 situation? 2) Will there be a change in the importance of socio-economic and travel-related variables between before and during COVID-19 for all travel modes?. The analytical tool adopted was the combination of the exploratory data analysis and the Artificial Neural Network (ANN) model. For both before and during the COVID-19 situation ANN models had been calibrated separately for each mode. The online survey (conducted between April 2020 and May 2020) for this research includes a comprehensive set of questions associated with individuals' travel behaviors, habits, and perceptions before, during, and after the outbreak, as well as their expectations about the future. Results revealed that the importance of variables for predicting the use of different modes at different stages of COVID-19 changed due to fear of virus spread, restrictions on travel, and affordability of different modes. During COVID-19 the most popular modes were walk and rickshaw. Socio-demographic variables including income, age, and gender were very important only for walk mode share at all stages of COVID-19. other travel-related variables such as trip purpose and travel cost have an important impact on mode choice decisions during-COVID-19. After-COVID-19 it can be anticipated that the walk, bicycle, and car mode shares would be moderately increased, and app-based service and motorcycle mode share would be slightly increased. The outcomes of this study can be applied to other developing countries with similar characteristics.

The ability of human encoded soluble proteins to convert into amyloid fibrils is now recognized as a generic phenomenon in several human illnesses. Typically, such disease causal proteins/peptides consist of aggregation-prone regions (APR) that make them susceptible to misfolding and assemble into highly ordered β-sheet rich fibrils, distinct from their native soluble state. Here, we show that the zika virus (ZIKV) consists of several such aggregation prone hotspots spread across its entire proteome. Using a combination of high-accuracy prediction tools, we identified APRs in both structural and non-structural proteins of ZIKV. Furthermore, we have experimentally validated the bioinformatic results by subjecting the ZIKV proteins and peptides to artificial aggregation inducing environment. Using a combination of dye-based assays (ThT and ANS) and microscopy techniques (HR-TEM and AFM), we further characterized the morphological features of amyloid-like fibrils. We found that Envelope domain III (EDIII) protein, NS1 β -roll peptide, membrane-embedded signal peptide 2K, and cytosolic region of NS4B protein to be highly aggregating in the experimental setup. Our findings also pave the way for an extensive and detailed functional analysis of these predicted APRs in the future to enhance our understanding of the role played by amyloids in the pathogenesis of flavivirus. Graphical Abstract

Background: To investigate the impact of the COVID-19 pandemic and infection prevention measures on children visiting emergency departments across Europe. Methods: Routine health data were extracted retrospectively from electronic patient records of children aged <16 years, presenting to 38 emergency departments (ED) in 16 European countries for the period January 2018 – May 2020, using predefined and standardised data domains. Observed and predicted numbers of ED attendances were calculated for the period February 2020 to May 2020. Poisson models and incidence rate ratios (IRR) were used to compare age groups, diagnoses, and outcomes. Findings: Reductions in paediatric ED attendances, hospital admissions and high triage urgencies were seen in all participating sites. ED attendances were relatively higher in countries with lower SARS-CoV-2 prevalence (incidence rate ratio (IRR) 2·62, 95% CI 2·19 to 3·13), and in children aged >12 months (12-<24 months IRR 0·89, 95% CI 0·86 to 0·92; 2-<5years IRR 0·84, 95% CI 0·82 to 0·87; 5-<12 years IRR 0·74, 95% CI 0·72 to 0·76; 12-<16 years IRR 0·74, 95% CI 0·71 to 0·77; vs. age <12 months as reference group). The impact on paediatric intensive care admissions (IRR 1·30, 95% CI 1·16 to 1·45) was not as great as the impact on general admissions. Lower triage urgencies were reduced more than higher triage urgencies (urgent triage IRR 1·10, 95% CI 1·08 to 1·12; emergent and very urgent triage IRR 1·53, 95% CI 1·49 to 1·57; vs. non-urgent triage category). Reductions were highest and sustained throughout the study period for children with communicable infectious diseases. Interpretation: Reductions in ED attendances were seen across Europe during the first COVID-19 lockdown period. More severely ill children continued to attend hospital more frequently compared to those with minor injuries and illnesses, although absolute numbers fell. Trial Registration Details: ISRCTN91495258. Funding Information: RGN was supported by National Institute of Health Research, award number ACL-2018-021- 007. Declaration of Interests: Authors have no conflict of interest to declare. Ethics Approval Statement: Following initial approval by the UK Health Research Authority, all participating sites obtained approval from their national/local institutional review boards. The need for individual patient informed consent was waived. Data sharing agreements were in place.

Zika virus (ZIKV) has been characterized as one of the potential pathogens and placed under future epidemic outbreaks by the WHO. However, lack of potential therapeutics can result in uncontrolled pandemic like other human pandemic viruses; therefore, prioritized effective therapeutics development has been recommended against ZIKV. In this context, the present study adopted the strategy to explore the lead compounds from Azadirachta indica against ZIKV via concurrent inhibition of the ZIKVpro and ZIKVRdRp proteins using molecular simulations. Initially, structure-based virtual screening of 44 bioflavonoids reported in Azadirachta indica against the crystal structures of targeted ZIKV proteins resulted in the identification of top four common bioflavonoids, viz. rutin, nicotiflorin, isoquercitrin, and hyperoside. These compounds showed substantial docking energy (-7.9 to -11.01 kcal/mol) and intermolecular interactions with essential residues of ZIKVpro (His51, Asp74, and Ser135) and ZIKVRdRp (Asp540, Ile799, and Asp665) by comparison to the reference compounds, O7N inhibitor (ZIKVpro) and Sofosbuvir inhibitor (ZIKVRdRp). Long interval molecular dynamics simulation (500 ns) on the selected docked poses reveals the stability of respective docked complexes contributed by intermolecular hydrogen bonds and hydrophobic interactions. The predicted complex stability was further supported by calculated end-point binding free energy using molecular mechanics generalized born surface area (MM/GBSA) method. Consequently, the identified common bioflavonoids are recommended as promising therapeutic inhibitors of ZIKVpro and ZIKVRdRp for further experimental assessment.

The E3 ligase TRIM7 has emerged as a critical player in viral infection and pathogenesis. A recent study found that TRIM7 inhibits human enteroviruses through ubiquitination and proteasomal degradation of viral 2BC protein by targeting the 2C moiety of 2BC protein. Here, we report the crystal structures of TRIM7 in complex with 2C, where the C-terminal region of 2C is inserted into a positively charged groove of the TRIM7 PRY-SPRY domain. Structure-guided biochemical studies revealed the C-terminus glutamine residue of 2C as the primary determinant for TRIM7 binding. Such a glutamine-end motif binding mechanism can be successfully extended to other substrates of TRIM7. More importantly, leveraged by this finding, we were able to identify norovirus and SARS-CoV-2 proteins, and physiological proteins, as new TRIM7 substrates. We further show that TRIM7 may function as a restriction factor to promote the degradation of the viral proteins of norovirus and SARS-CoV-2, thereby restoring the Type I interferon immune response and inhibiting viral infection. Several crystal structures of TRIM7 in complex with SARS-CoV-2 proteins are also determined, and a conserved C-terminus glutamine-specific interaction is observed. These findings unveil a common recognition mode by TRIM7, providing the foundation for further mechanistic characterization of antiviral and cellular functions of TRIM7.

ABSTRACT Biological control of mosquito vectors using the insect-specific bacteria Wolbachia is an emerging strategy for the management of human arboviral diseases. We recently described the development of a strain of Ae. aegypti infected with the Wolbachia strain w AlbB (referred to as the w AlbB2-F4 strain) through simple backcrossing of wild type Australian mosquitoes with a w AlbB infected Ae. aegypti strain from the USA. Field releases of male w AlbB2-F4 mosquitoes resulted in the successful suppression of a wild population of mosquitoes in the trial sites by exploiting the strains Wolbachia- induced cytoplasmic incompatibility. We now demonstrate that the strain is resistant to infection by dengue and Zika viruses and is genetically similar to endemic Queensland populations. There was a fourfold reduction in the proportion of w AlbB2-F4 mosquitoes that became infected following a blood meal containing dengue 2 virus (16.7%) compared to wild type mosquitoes (69.2%) and a 6-7 fold reduction in the proportion of w AlbB2-F4 mosquitoes producing virus in saliva following a blood meal containing an epidemic strain of Zika virus (8.7% in comparison to 58.3% in wild type mosquitoes). Restriction-site Associated DNA (RAD) sequencing revealed that w AlbB2-F4 mosquitoes have > 98% Australian ancestry, confirming the successful introduction of the w AlbB2 infection into the Australian genomic background through backcrossing. Genotypic and phenotypic analyses showed the w AlbB2-F4 strain retains the insecticide susceptibility phenotype and genotype of the Australian mosquitoes. We demonstrate that the Wolbachia w AlbB2-F4, in addition to being suitable for suppression programs, can be effective in population replacement programs given its high inhibition of virus infection in mosquitoes. The ease at which a target mosquito population can be transfected with w AlbB2, while retaining genotypes and phenotypes of the target population, shows the robustness of this strain as a biocontrol agent against the Ae. aegypti mosquito itself as well as the pathogens it transmits. IMPORTANCE Epidemics of arthopod-borne virus (arbovirus) diseases affect millions of people and are becoming more frequent and widespread. A successful strategy to control these diseases is by infecting mosquito populations with benign, insect-specific Wolbachia bacteria that render mosquitoes refractory to infection with pathogenic arboviruses. Here we show that a strain of the major mosquito vector Ae. aegypti that was infected with Wolbachia following a simple back-cross mating procedure is refractory to infection with dengue and Zika viruses. Importantly, the genetic background of the strain is equivalent to the target population, which is important for persistence of the strain and regulatory approval.

The World Health Organization (WHO) notifies the global community about disease outbreaks through the Disease Outbreak News (DON). These online reports tell important stories about both outbreaks themselves and the high-level decision making that governs information sharing during public health emergencies. However, they have been used only minimally in global health scholarship to date. Here, we collate all 2,789 of these reports from their first use through the start of the Covid-19 pandemic (January 1996 to December 2019), and develop an annotated database of the subjective and often inconsistent information they contain. We find that these reports are dominated by a mix of persistent worldwide threats (particularly influenza and cholera) and persistent epidemics (like Ebola virus disease in Africa or MERS-CoV in the Middle East), but also document important periods in history like the anthrax bioterrorist attacks at the turn of the century, the spread of chikungunya and Zika virus to the Americas, or even recent lapses in progress towards polio elimination. We present three simple vignettes that show how researchers can use these data to answer both qualitative and quantitative questions about global outbreak dynamics and public health response. However, we also find that the retrospective value of these reports is visibly limited by inconsistent reporting (e.g., of disease names, case totals, mortality, and actions taken to curtail spread). We conclude that sharing a transparent rubric for which outbreaks are considered reportable, and adopting more standardized formats for sharing epidemiological metadata, might help make the DON more useful to researchers and policymakers.

Countermeasures against Zika virus (ZIKV), including vaccines, are frequently tested in nonhuman primates (NHP). Macaque models are important for understanding how ZIKV infections impact human pregnancy due to similarities in placental development. The lack of consistent adverse pregnancy outcomes in ZIKV-affected pregnancies poses a challenge in macaque studies where group sizes are often small (4-8 animals). Studies in small animal models suggest that African-lineage Zika viruses can cause more frequent and severe fetal outcomes. No adverse outcomes were observed in macaques inoculated with a low dose of African-lineage ZIKV at gestational day (GD) 45. Here, we inoculate eight pregnant rhesus macaques with a higher dose of African-lineage ZIKV at GD 45 to test the hypothesis that adverse pregnancy outcomes are dose-dependent. Three of eight pregnancies ended prematurely with fetal death. ZIKV was detected in both fetal and placental tissues from all cases of early fetal loss. Further refinements of this challenge system (e.g., varying the dose and timing of infection) could lead to an even more consistent, unambiguous fetal loss phenotype for assessing ZIKV countermeasures in pregnancy. These data demonstrate that high-dose inoculation with African-lineage ZIKV causes pregnancy loss in macaques and also suggest that ZIKV-induced first trimester pregnancy could be strain-specific.

Placental pathology can identify characteristic features of specific infectious pathogens. The histopathology of acute SARS-CoV-2 placental infection and exposure without infection has been well described. However, whether the characteristic placental pathology persists after the acute phase of the infection is less clear. We retrospectively identified 67 COVID-19 recovered pregnant patients who had placental pathology available. After reviewing the gross and histopathology we categorized the findings and studied the placentas for evidence of chronic infection by immunohistochemistry for the Spike protein of the virus. We found these placentas showed significantly increased prevalence of maternal vascular malperfusion and trend towards increased prevalence of fetal vascular malperfusion pathologies when compared to a control group of placentas examined for the sole indication of maternal GBS colonization. None of the COVID-19 recovered placentas showed expression of the viral protein, therefore we found no evidence of persistent infection of the placenta in women with a history of COVID-19 during their pregnancy... We conclude that recovery from a SARS-CoV-2 infection during pregnancy puts the pregnancy at risk for specific pathology.

Bloodstream infection caused by antimicrobial resistance pathogens is a global concern because it is difficult to treat with conventional therapy. Here we report scavenger magnetic nanoparticles enveloped by nanovesicles derived from blood cells (MNVs), which magnetically eradicate an extreme range of pathogens in an extracorporeal circuit. We quantitatively reveal that glycophorin A and complement receptor (CR) 1 on red blood cell (RBC)-MNVs predominantly capture human fecal bacteria, carbapenem-resistant (CR) E. coli , and extended-spectrum beta-lactamases-positive (ESBL-positive) E. coli , vancomycin-intermediate S. aureus (VISA), endotoxins, and proinflammatory cytokines in human blood. Additionally, CR3 and CR1 on white blood cell-MNVs mainly contribute to depleting the virus envelope proteins of Zika, SARS-CoV-2, and their variants in human blood. Supplementing opsonins into the blood significantly augments the pathogen removal efficiency due to its combinatorial interactions between pathogens and CR1 and CR3 on MNVs. The extracorporeal blood cleansing enables full recovery of lethally infected rodent animals within seven days by treating them twice in series. We also validate that parameters reflecting immune homeostasis, such as blood cell counts, cytokine levels, and transcriptomics changes, are restored in blood of the fatally infected rats after treatment.

This contribution marks a dual milestone at the intersection of public health law and JLME: my 50th publication of a substantive manuscript in the 50th anniversary of the Journal of Law, Medicine & Ethics in 2022. In recognition of these coinciding landmarks, this piece for JLME features observations and reflections of the field of public health law and policy.

Artificial intelligence (AI) has been applied widely in our daily lives in a variety of ways with numerous success stories. AI has also contributed to dealing with the coronavirus disease (COVID-19) pandemic, which has been happening around the globe. This paper presents a survey of AI methods being used in various applications in the fight against the COVID-19 outbreak and outlines the crucial role of AI research in this unprecedented battle. We touch on areas where AI plays as an essential component, from medical image processing, data analytics, text mining and natural language processing, the Internet of Things, to computational biology and medicine. A summary of COVID-19 related data sources that are available for research purposes is also presented. Research directions on exploring the potential of AI and enhancing its capability and power in the pandemic battle are thoroughly discussed. We identify 13 groups of problems related to the COVID-19 pandemic and highlight promising AI methods and tools that can be used to address these problems. It is envisaged that this study will provide AI researchers and the wider community with an overview of the current status of AI applications, and motivate researchers to harness AI's potential in the fight against COVID-19.