Molecular Characterization of Influenza A/H3N2 Virus Isolated from Indonesian Hajj and Umrah Pilgrims 2013 to 2014.

The Hajj and Umrah are the annual mass gatherings of Muslims in Saudi Arabia and increase the transmission risk of acute respiratory infection. This study describes influenza infection among pilgrims upon arrival in Indonesia and the genetic characterization of imported influenza A/H3N2 virus. In total, 251 swab samples with influenza-like illness were tested using real-time RT-PCR for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and influenza viruses. Complete sequences of influenza A/H3N2 HA and NA genes were obtained using DNA sequencing and plotted to amino acid and antigenicity changes. Phylogenetic analysis was performed using a neighbour-joining method including the WHO vaccine strains and influenza A/H3N2 as references. The real-time RT-PCR test detected 100 (39.5%) samples positive with influenza with no positivity of MERS-CoV. Mutations in the HA gene were mainly located within the antigenic sites A, B, and D, while for the NA gene, no mutations related to oseltamivir resistance were observed. Phylogenetic analysis revealed that these viruses grouped together with clades 3C.2 and 3C.3; however, they were not closely grouped with the WHO-recommended vaccine (clades 3C.1). Sequences obtained from Hajj and Umrah pilgrims were also not grouped together with viruses from Middle East countries but clustered according to years of collection. This implies that the influenza A/H3N2 virus mutates continually across time.

A pioneering effort to give voices to indigenous psychology in Southeast Asia

For this special issue, eight manuscripts were accepted. The contributions showcase a range of indigenous psychological concepts and practices in the Southeast Asia (SEA) region. These include the Filipino values of "kapwa" (shared identity) and Indonesian values of "Mangan ora mangan sing penting kumpul" and "Dalihan Na Tolu" (both referring to the importance of kinship) in relation to one's preferred sources of social supports, Filipino coping strategies of "diskarte" (resourcefulness) and "bayanihan" (mutual, collective support) in managing mental health challenges, the "Sikolohiyang Pilipino" (Filipino Psychology) movement in the Philippines, the "kiasu" mindset (fear of losing out), and traditional Chinese healing practices of "dang-ki" (consultations with a divine deity) in Singapore. The contributions all address sustainable development goals in SEA communities, such as mental health challenges and practices in Indonesia, the Philippines and Singapore;the marginalization of ethnic minority groups;effects of postcolonialism in the Philippines and Malaysia;and issues of urbanization, deforestation, and climate change in Malaysia. Some assessed specifically the effects of the COVID-19 pandemic on SEA populations' well-being, including comparisons of COVID-19 stigma and quality of life in Indonesia and the Philippines, the associated factors of self-harm and suicide ideation among Chinese Indonesians, and the mental health challenges faced and coping strategies utilized by doctors working in rural areas in the Philippines. (PsycInfo Database Record (c) 2023 APA, all rights reserved)

Wearable devices for anxiety & depression: A scoping review.

Background: The rates of mental health disorders such as anxiety and depression are at an all-time high especially since the onset of COVID-19, and the need for readily available digital health care solutions has never been greater. Wearable devices have increasingly incorporated sensors that were previously reserved for hospital settings. The availability of wearable device features that address anxiety and depression is still in its infancy, but consumers will soon have the potential to self-monitor moods and behaviors using everyday commercially-available devices. Objective: This study aims to explore the features of wearable devices that can be used for monitoring anxiety and depression. Methods: Six bibliographic databases, including MEDLINE, EMBASE, PsycINFO, IEEE Xplore, ACM Digital Library, and Google Scholar were used as search engines for this review. Two independent reviewers performed study selection and data extraction, while two other reviewers justified the cross-checking of extracted data. A narrative approach for synthesizing the data was utilized. Results: From 2408 initial results, 58 studies were assessed and highlighted according to our inclusion criteria. Wrist-worn devices were identified in the bulk of our studies (n = 42 or 71%). For the identification of anxiety and depression, we reported 26 methods for assessing mood, with the State-Trait Anxiety Inventory being the joint most common along with the Diagnostic and Statistical Manual of Mental Disorders (n = 8 or 14%). Finally, n = 26 or 46% of studies highlighted the smartphone as a wearable device host device. Conclusion: The emergence of affordable, consumer-grade biosensors offers the potential for new approaches to support mental health therapies for illnesses such as anxiety and depression. We believe that purposefully-designed wearable devices that combine the expertise of technologists and clinical experts can play a key role in self-care monitoring and diagnosis.

Analyzing Jo-Ha-Kyu Mechanism in Japanese Traditional Performing Art Ningyo Joruri

IoT devices that connect people without physical contact become more and more important after the COVID-19 impact. However, strange appearances and movements performed by IoT devices (interactive humanoid robots) cause human discomfort, so-called the uncanny valley, preventing widespread use of humanoid IoT devices. On the contrary, a Japanese traditional performing art named Ningyo Joruri (puppet theater) is recognized as a UNESCO intangible cultural heritage, and the sophisticated puppet motions and its unique music style somehow can avoid causing human discomfort even if the appearance of puppets is close enough to humans. One of the most important factors in empathizing humans with the puppet without uncomfortable is the modulation technique of both music tempo and motion speed known as Jo-Ha-Kyu. In this study, we analyzed Ningyo Joruri based on the Jo-Ha-Kyu mechanism, which is an art concept adopted in the puppet theater to interact with audiences according to modulation of the tempo. First, we obtained puppet movements using motion capture systems with the music. Second, we detected the changing tempo in Ningyo Joruri using the deep learning method to demonstrate the Jo-Ha-Kyu mechanism quantitatively. Finally, we showed the correlation of Jo-Ha-Kyu between Ningyo Joruri music and puppet manipulation techniques in the frequency domain using the Hilbert Huang transform. Our results revealed that low-frequency movements play an important role in synchronizing motion to the tempo of corresponding music, presenting novel knowledge to motion designers for humanoid robots IoT devices. © 2022 IEEE.

Use of radiolabeled hyaluronic acid for preclinical assessment of inflammatory injury and acute respiratory distress syndrome.

Acute respiratory distress syndrome (ARDS) is accompanied by a dramatic increase in lung hyaluronic acid (HA), leading to a dose-dependent reduction of pulmonary oxygenation. This pattern is associated with severe infections, such as COVID-19, and other important lung injury etiologies. HA actively participates in molecular pathways involved in the cytokine storm of COVID-19-induced ARDS. The objective of this study was to evaluate an imaging approach of radiolabeled HA for assessment of dysregulated HA deposition in mouse models with skin inflammation and lipopolysaccharide (LPS)-induced ARDS using a novel portable intensified Quantum Imaging Detector (iQID) gamma camera system. METHODS: HA of 10 kDa molecular weight (HA10) was radiolabeled with 125I and 99mTc respectively to produce [125I]I-HA10 and [99mTc]Tc-HA10, followed by comparative studies on stability, in vivo biodistribution, and uptake at inflammatory skin sites in mice with 12-O-tetradecanoylphorbol-13-acetate (TPA)-inflamed ears. [99mTc]Tc-HA10 was used for iQID in vivo dynamic imaging of mice with ARDS induced by intratracheal instillation of LPS. RESULTS: [99mTc]Tc-HA10 and [125I]I-HA10 had similar biodistribution and localization at inflammatory sites. [99mTc]Tc-HA10 was shown to be feasible in measuring skin injury and monitoring skin wound healing. [99mTc]Tc-HA10 dynamic pulmonary images yielded good visualization of radioactive uptake in the lungs. There was significantly increased lung uptake and slower lung washout in mice with LPS-induced ARDS than in control mice. Postmortem biodistribution measurement of [99mTc]TcHA10 (%ID/g) was 11.0 ± 3.9 vs. 1.3 ± 0.3 in the ARDS mice (n = 6) and controls (n = 6) (P < 0.001), consistent with upregulated HA expression as determined by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) staining. CONCLUSIONS: [99mTc]Tc-HA10 is promising as a biomarker for evaluating HA dysregulation that contributes to pulmonary injury in ARDS. Rapid iQID imaging of [99mTc]Tc-HA10 clearance from injured lungs may provide a functional template for timely assessment and quantitative monitoring of pulmonary pathophysiology and intervention in ARDS.

Accidental poisoning among children in a regional hospital in northern KwaZulu-Natal before and during the COVID-19 pandemic

Introduction. Poisoning has been reported as the fifth most common cause of injury-related deaths in children <5 years worldwide. Little is known about accidental poisoning among children in the regional setting. During the COVID-19 pandemic, lockdown measures may have increased home-based unintentional poisoning. Objectives. To determine the frequency, outcome and type of accidental poisoning in children admitted to a regional hospital and compare cases before and during the COVID-19 pandemic. Methods. A review of admissions to Queen Nandi Regional Hospital in Empangeni was performed to document cases of accidental poisoning >2 years. Equal periods during 2019 and 2020 (April to December) were compared. Children <13 years were included. Age, sex, date of admission, death, survival and type of poisoning were collected. Results. Accidental poisoning made up a small proportion of the total admissions (n=252/5 071;4.97%) with a low case fatality rate (0.40%). Boys made up the majority (n=132/252, 52.38%). Most were <5 years (n=220/252, 87.30%,). Medicines (n=114/252, 45.24%), hydrocarbons (n=61/252, 24.21%) and pesticides (n=26/252, 10.32%) were the main types of poisoning. Domestic cleaner, sanitiser or disinfectant-related admissions were significantly increased during the pandemic (p=0.020). Conclusion. Accidental poisoning commonly occurs in younger children. Medicines, hydrocarbons and pesticides make up the majority of cases. Domestic cleaner, hand sanitiser and disinfectant ingestion increased during the COVID pandemic. Future research involving primary care facilities and risk factors related to poisoning should be investigated.

Bioinformatics analysis on structure and function and expression vector construction of SARS-CoV-2 related protein TMPRSS2

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and function of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular cloning technology.The homology, functional sites, subcellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly;TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L * mol~1 * cm"1 , and the half-life is 30 h;TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addition, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolutionary analysis showed that the highest sequence consistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ideas for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein. Copyright © 2022 Publication Centre of Anhui Medical University. All rights reserved.

Does a Gluten-Free Diet Affect BMI and Glycosylated Hemoglobin in Children and Adolescents with Type 1 Diabetes and Asymptomatic Celiac Disease? A Meta-Analysis and Systematic Review.

Background: Children diagnosed with type 1 diabetes mellitus (T1DM) are more prone to having celiac disease (CD) than the normal population. Moreover, patients with this dual diagnosis who are also on a diabetic and gluten-free diet (GFD) risk faltering growth and uncontrolled blood glucose levels. This review aims to assess the efficacy and effectiveness of managing patients with T1DM screened for CD with GFD to prevent complications associated with these chronic pathologies in childhood and adulthood. Materials and Methods: We abided by the PRISMA guidelines in this meta-analysis and used multiple databases and search engines. We included case-control studies. The primary outcomes were changes in the standard deviation score, body mass index (SDS BMI), and glycosylated hemoglobin (HA1C) after being on a GFD for at least twelve months. Results: The pooled data from the six studies included showed that there was neither a statistically significant difference in the mean SDS BMI (-0.28 (95% CI -0.75, 0.42)) (p = 0.24) nor in the mean of HA1C (mean -0.07 (95% CI -0.44, 0.30)) (p = 0.36) for the same group. HDL cholesterol improved significantly in patients on a strict GFD (p < 0.01). Conclusions: In children with T1DM and asymptomatic CD, being on a GFD had no significant effect on BMI or HA1C. However, it can have a protective effect on the other complications found in both chronic pathologies.

Elucidating the enhanced binding affinity of a double mutant SP-D with trimannose on the influenza A virus using molecular dynamics.

Surfactant protein D (SP-D) is an essential component of the human pulmonary surfactant system, which is crucial in the innate immune response against glycan-containing pathogens, including Influenza A viruses (IAV) and SARS-CoV-2. Previous studies have shown that wild-type (WT) SP-D can bind IAV but exhibits poor antiviral activities. However, a double mutant (DM) SP-D consisting of two point mutations (Asp325Ala and Arg343Val) inhibits IAV more potently. Presently, the structural mechanisms behind the point mutations' effects on SP-D's binding affinity with viral surface glycans are not fully understood. Here we use microsecond-scale, full-atomistic molecular dynamics (MD) simulations to understand the molecular mechanism of mutation-induced SP-D's higher antiviral activity. We find that the Asp325Ala mutation promotes a trimannose conformational change to a more stable state. Arg343Val increases the binding with trimannose by increasing the hydrogen bonding interaction with Glu333. Free energy perturbation (FEP) binding free energy calculations indicate that the Arg343Val mutation contributes more to the increase of SP-D's binding affinity with trimannose than Asp325Ala. This study provides a molecular-level exploration of how the two mutations increase SP-D binding affinity with trimannose, which is vital for further developing preventative strategies for related diseases.

Kinetic of the Antibody Response Following AddaVax-Adjuvanted Immunization with Recombinant Influenza Antigens.

Notwithstanding the current SARS-CoV-2 pandemic, influenza virus infection still represents a global health concern in terms of hospitalizations and possible pandemic threats. The objective of next-generation influenza vaccines is not only to increase the breadth of response but also to improve the elicitation of an effective and robust immune response, especially in high-risk populations. To achieve this second objective, the administration of adjuvanted influenza vaccines has been considered. In this regard, the monitoring and characterization of the antibody response associated with the administration of adjuvanted vaccines has been evaluated in this study in order to shed light on the kinetic, magnitude and subclass usage of antibody secreting cells (ASCs) as well as of circulating antigen-specific serum antibodies. Specifically, we utilized the DBA/2J mouse model to assess the kinetic, magnitude and IgG subclass usage of the antibody response following an intramuscular (IM) or intraperitoneal (IP) immunization regimen with AddaVax-adjuvanted bivalent H1N1 and H3N2 computationally optimized broadly reactive antigen (COBRA) influenza recombinant hemagglutinins (rHAs). While the serological evaluation revealed a homogeneous kinetic of the antibody response, the detection of the ASCs through a FluoroSpot platform revealed a different magnitude, subclass usage and kinetic of the antigen-specific IgG secreting cells peaking at day 5 and day 9 following the IP and IM immunization, respectively.

Biopolymeric sustainable materials and their emerging applications

Advancements in polymer science and engineering have helped the scientific community to shift its attention towards the use of environmentally benign materials for reducing the environmental impact of conventional synthetic plastics. Biopolymers are environmentally benign, chemically versatile, sustainable, biocompatible, biodegradable, inherently functional, and ecofriendly materials that exhibit tremendous potential for a wide range of applications including food, electronics, agriculture, textile, biomedical, and cosmetics. This review also inspires the researchers toward more consumption of biopolymer-based composite materials as an alternative to synthetic composite materials. Herein, an overview of the latest knowledge of different natural- and synthetic-based biodegradable polymers and their fiber-reinforced composites is presented. The review discusses different degradation mechanisms of biopolymer-based composites as well as their sustainability aspects. This review also elucidates current challenges, future opportunities, and emerging applications of biopolymeric sustainable composites in numerous engineering fields. Finally, this review proposes biopolymeric sustainable materials as a propitious solution to the contemporary environmental crisis. © 2022 Elsevier Ltd.

Preliminary Observations and Impact in Japan of the Tsunami Caused by the Tonga Volcanic Eruption on January 15, 2022.

The tsunami caused by the Tonga submarine volcanic eruption that occurred at 13:15 Japan Time (JST) on January 15, 2022, exposed a blind spot in Japan's tsunami monitoring and warning system, which was established in 1952 for local tsunamis and expanded to distant tsunamis after the 1960 Chile tsunami. This paper summarizes how the warning system responded to the unprecedented tsunami, the actual evacuation process, and the damage it caused in Japan. Initially, the tsunami from the volcanic eruption was expected to arrive at approximately midnight with amplitudes of less than 20 cm. However, a series of short waves arrived at approximately 21:00, a few hours earlier than expected. The early arrival of these sea waves coincided with a rapid increase in atmospheric pressure; then, the short-period component was predominant, and the wave height was amplified while forming wave groups. After a 1.2 m tsunami was observed in Amami City in southern Japan at 23:55 JST, the Japan Meteorological Agency issued a tsunami warning/advisory. The tsunami continued, and all advisories were cleared at 14:00 JST on January 16. Information about this tsunami and the response to it are summarized here, including the characteristics and issues of the actual tsunami evacuation situation in each region. There were no casualties, but the issues that emerged included difficulty evacuating on a winter night and traffic congestion due to evacuation by car and under the conditions of the COVID-19 pandemic. In the coastal area, damage to fishing boats and aquaculture facilities was reported due to the flow of the tsunami. In addition, damage to aquaculture facilities, including those producing oysters, scallops, seaweed and other marine products, decreased the supply of marine products, and the economic impact is likely to increase in the future.

Prospects of NIR fluorescent nanosensors for green detection of SARS-CoV-2.

The pandemic of the novel coronavirus disease 2019 (COVID-19) is continuously causing hazards for the world. Effective detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can relieve the impact, but various toxic chemicals are also released into the environment. Fluorescence sensors offer a facile analytical strategy. During fluorescence sensing, biological samples such as tissues and body fluids have autofluorescence, giving false-positive/negative results because of the interferences. Fluorescence near-infrared (NIR) nanosensors can be designed from low-toxic materials with insignificant background signals. Although this research is still in its infancy, further developments in this field have the potential for sustainable detection of SARS-CoV-2. Herein, we summarize the reported NIR fluorescent nanosensors with the potential to detect SARS-CoV-2. The green synthesis of NIR fluorescent nanomaterials, environmentally compatible sensing strategies, and possible methods to reduce the testing frequencies are discussed. Further optimization strategies for developing NIR fluorescent nanosensors to facilitate greener diagnostics of SARS-CoV-2 for pandemic control are proposed.

Genetic Analysis of Influenza A/H1N1pdm Strains Isolated in Bangladesh in Early 2020.

Influenza is one of the most common respiratory virus infections. We analyzed hemagglutinin (HA) and neuraminidase (NA) gene segments of viruses isolated from influenza patients who visited Evercare Hospital Dhaka, Bangladesh, in early 2020 immediately before the coronavirus disease 2019 (COVID-19) pandemic. All of them were influenza virus type A (IAV) H1N1pdm. Sequence analysis of the HA segments of the virus strains isolated from the clinical specimens and the subsequent phylogenic analyses of the obtained sequences revealed that all of the H1N1pdm recent subclades 6B.1A5A + 187V/A, 6B.1A5A + 156K, and 6B.1A5A + 156K with K209M were already present in Bangladesh in January 2020. Molecular clock analysis results suggested that the subclade 6B.1A5A + 156K emerged in Denmark, Australia, or the United States in July 2019, while subclades 6B.1A5A + 187V/A and 6B.1A5A + 156K with K209M emerged in East Asia in April and September 2019, respectively. On the other hand, sequence analysis of NA segments showed that the viruses lacked the H275Y mutation that confers oseltamivir resistance. Since the number of influenza cases in Bangladesh is usually small between November and January, these results indicated that the IAV H1N1pdm had spread extremely rapidly without acquiring oseltamivir resistance during a time of active international flow of people before the COVID-19 pandemic.

Development of a hybrid alphavirus-SARS-CoV-2 pseudovirion for rapid quantification of neutralization antibodies and antiviral drugs.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein (S)-pseudotyped viruses are commonly used for quantifying antiviral drugs and neutralizing antibodies. Here, we describe the development of a hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) pseudovirion, which is a non-replicating SARS-CoV-2 virus-like particle composed of viral structural proteins (S, M, N, and E) and an RNA genome derived from a fast-expressing alphaviral vector. We validated Ha-CoV-2 for rapid quantification of neutralization antibodies, antiviral drugs, and viral variants. In addition, as a proof of concept, we used Ha-CoV-2 to quantify the neutralizing antibodies from an infected and vaccinated individual and found that the one-dose vaccination with Moderna mRNA-1273 greatly increased the anti-serum titer by approximately 6-fold. The post-vaccination serum can neutralize all nine variants tested. These results demonstrate that Ha-CoV-2 can be used as a robust platform for the rapid quantification of neutralizing antibodies against SARS-CoV-2 and its emerging variants.

Corona Discharge and Field Electron Emission in Ambient Air Using a Sharp Metal Needle: Formation and Reactivity of CO3 -• and O2 -•.

CO3 -• and O2 -• are known to be strong oxidizing reagents in biological systems. CO3 -• in particular can cause serious damage to DNA and proteins by H• abstraction reactions. However, H• abstraction of CO3 -• in the gas phase has not yet been reported. In this work we report on gas-phase ion/molecule reactions of CO3 -• and O2 -• with various molecules. CO3 -• was generated by the corona discharge of an O2 reagent gas using a cylindrical tube ion source. O2 -• was generated by the application of a 15 kHz high frequency voltage to a sharp needle in ambient air at the threshold voltage for the appearance of an ion signal. In the reactions of CO3 -•, a decrease in signal intensities of CO3 -• accompanied by the simultaneous increase of that of HCO3 - was observed when organic compounds with H-C bond energies lower than ∼100 kcal mol-1 such as n-hexane, cyclohexane, methanol, ethanol, 1-propanol, 2-propanol, and toluene were introduced into the ion source. This clearly indicates the occurrence of H• abstraction. O2 -• abstracts H+ from acid molecules such as formic, acetic, trifluoroacetic, nitric and amino acids. Gas-phase CO3 -• may play a role as a strong oxidizing reagent as it does in the condensed phase. The major discharge product CO3 -• in addition to O2 -•, O3, and NO x • that are formed in ambient air may cause damage to biological systems.

Performance evaluation of virus concentration methods for implementing SARS-CoV-2 wastewater based epidemiology emphasizing quick data turnaround.

Wastewater based epidemiology (WBE) has drawn significant attention as an early warning tool to detect and predict the trajectory of COVID-19 cases in a community, in conjunction with public health data. This means of monitoring for outbreaks has been used at municipal wastewater treatment centers to analyze COVID-19 trends in entire communities, as well as by universities and other community living environments to monitor COVID-19 spread in buildings. Sample concentration is crucial, especially when viral abundance in raw wastewater is below the threshold of detection by RT-qPCR analysis. We evaluated the performance of a rapid ultrafiltration-based virus concentration method using InnovaPrep Concentrating Pipette (CP) Select and compared this to the established electronegative membrane filtration (EMF) method. We evaluated sensitivity of SARS-CoV-2 quantification, surrogate virus recovery rate, and sample processing time. Results suggest that the CP Select concentrator is more efficient at concentrating SARS-CoV-2 from wastewater compared to the EMF method. About 25% of samples that tested negative when concentrated with the EMF method produced a positive signal with the CP Select protocol. Increased recovery of the surrogate virus control using the CP Select confirms this observation. We optimized the CP Select protocol by adding AVL lysis buffer and sonication, to increase the recovery of virus. Sonication increased Bovine Coronavirus (BCoV) recovery by 19%, which seems to compensate for viral loss during centrifugation. Filtration time decreases by approximately 30% when using the CP Select protocol, making this an optimal choice for building surveillance applications where quick turnaround time is necessary.

Recombinant myxoma virus infection associated with high mortality in rabbit farming (Oryctolagus cuniculus).

Myxomatosis is an emergent disease in the Iberian hare, having been considered a rabbit disease for decades. Genome sequencing of the strains obtained from Iberian hares with myxomatosis showed these to be distinct from the classical ones that circulated in rabbits since the virus introduction in Europe, in 1952. The main genomic difference in this natural recombinant hare myxoma virus (ha-MYXV) is the presence of an additional 2.8 kb region disrupting the M009L gene and adding a set of genes homologous to the myxoma virus (MYXV) genes M060R, M061R, M064R, M065R and M066R originated in Poxviruses. After the emergence of this recombinant virus (ha-MYXV) in hares, in the summer of 2019, the ha-MYXV was not detected in rabbit surveys, suggesting an apparent species segregation with the MYXV classic strains persistently circulating in rabbits. Recently, a group of six unvaccinated European rabbits (Oryctolagus cuniculus cuniculus) from a backyard rabbitry in South Portugal developed signs of myxomatosis (anorexia, dyspnoea, oedema of eyelids, head, ears, external genitals and anus, and skin myxomas in the base of the ears). Five of them died within 24-48 hr of symptom onset. Molecular analysis revealed that only the recombinant MYXV was present. This is the first documented report of a recombinant hare myxoma virus in farm rabbits associated with high mortality, which increases the concern for the future of both the Iberian hare and wild rabbits and questions the safety of the rabbit industry. This highlights the urgent need to evaluate the efficacy of available vaccines against this new MYXV.

S-Acylation of Proteins of Coronavirus and Influenza Virus: Conservation of Acylation Sites in Animal Viruses and DHHC Acyltransferases in Their Animal Reservoirs.

Recent pandemics of zoonotic origin were caused by members of coronavirus (CoV) and influenza A (Flu A) viruses. Their glycoproteins (S in CoV, HA in Flu A) and ion channels (E in CoV, M2 in Flu A) are S-acylated. We show that viruses of all genera and from all hosts contain clusters of acylated cysteines in HA, S and E, consistent with the essential function of the modification. In contrast, some Flu viruses lost the acylated cysteine in M2 during evolution, suggesting that it does not affect viral fitness. Members of the DHHC family catalyze palmitoylation. Twenty-three DHHCs exist in humans, but the number varies between vertebrates. SARS-CoV-2 and Flu A proteins are acylated by an overlapping set of DHHCs in human cells. We show that these DHHC genes also exist in other virus hosts. Localization of amino acid substitutions in the 3D structure of DHHCs provided no evidence that their activity or substrate specificity is disturbed. We speculate that newly emerged CoVs or Flu viruses also depend on S-acylation for replication and will use the human DHHCs for that purpose. This feature makes these DHHCs attractive targets for pan-antiviral drugs.

Identification of the SHREK Family of Proteins as Broad-Spectrum Host Antiviral Factors.

Mucins and mucin-like molecules are highly glycosylated, high-molecular-weight cell surface proteins that possess a semi-rigid and highly extended extracellular domain. P-selectin glycoprotein ligand-1 (PSGL-1), a mucin-like glycoprotein, has recently been found to restrict HIV-1 infectivity through virion incorporation that sterically hinders virus particle attachment to target cells. Here, we report the identification of a family of antiviral cellular proteins, named the Surface-Hinged, Rigidly-Extended Killer (SHREK) family of virion inactivators (PSGL-1, CD43, TIM-1, CD34, PODXL1, PODXL2, CD164, MUC1, MUC4, and TMEM123) that share similar structural characteristics with PSGL-1. We demonstrate that SHREK proteins block HIV-1 infectivity by inhibiting virus particle attachment to target cells. In addition, we demonstrate that SHREK proteins are broad-spectrum host antiviral factors that block the infection of diverse viruses such as influenza A. Furthermore, we demonstrate that a subset of SHREKs also blocks the infectivity of a hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) pseudovirus. These results suggest that SHREK proteins may be a part of host innate immunity against enveloped viruses.