title: Mapping the evolutionary trajectories of morbilliviruses: what, where and whither

text: Morbilliviruses are pathogens of humans and other animals. Live attenuated morbillivirus vaccines have been used to end endemic transmission of measles virus (MV) in many parts of the developed world and to eradicate rinderpest virus. Entry is mediated by two different receptors which govern virus lymphotropism and epitheliotropism. Morbillivirus transmissibility is unparalleled and MV represents the most infectious human pathogen on earth. Their evolutionary origins remain obscure and their potential for adaption to new hosts is poorly understood. It has been suggested that MV could be eradicated. Therefore it is imperative to dissect barriers which restrict cross species infections. This is important as ecological studies identify novel morbilliviruses in a vast number of small mammals and carnivorous predators.


title: Influence of weaning date and late gestation supplementation on beef system productivity I: animal performance()

text: A 4-yr experiment examined how weaning date and prepartum nutrition affected productivity in a spring (March and April) calving system. Crossbred beef cows (479 ± 59 kg, n = 144) were used in a completely randomized design with a 2 × 4 factorial treatment arrangement: 1) cows were weaned in early October or early December; and 2) during late gestation cows were fed on a dry matter basis a 32% crude protein supplement at 0, 0.41, or 0.82 kg/cow/d on dormant upland range or grazed corn residue without supplement. Cow body condition score (BCS) was affected (P ≤ 0.01) by treatment prior to parturition and breeding but was similar (P > 0.27) among all treatments in October. Dams on a higher nutritional plane during winter treatment had greater (P < 0.01) BCS and body weight (BW) prior to parturition and breeding. Subsequent pregnancy rates (88% to 97%) were not influenced (P > 0.76) by weaning date, but tended (P = 0.10) to be lower for cows grazing winter range without supplement. Calves born to dams grazing winter range without supplement had lower (P < 0.01) BW in October and adjusted weaning BW. Pre-breeding BW of heifers weaned in December born to cows grazing winter range without supplement was lower (P < 0.01) than contemporaries born to cows in all other treatment combinations. However, postweaning (0.48 kg/d) and postbreeding (0.42 kg/d) average daily gain, percentage cycling before breeding (33%), and pregnancy rate (81%) was similar (P > 0.12). Within weaning date, steers born to cows grazing winter range without supplement had lower (P < 0.05) hot carcass weight (HCW) than contemporaries born to cows grazing corn residue. Cows weaned in December had decreased BW and BCS but similar pregnancy rates as cows weaned in October. Weaning date and dam maternal nutrition had minimal impact on heifer progeny pregnancy rate. Steer progeny born to dams on a higher nutritional plane had similar HCW at slaughter when adjusted to equal fat thickness.


title: Consensus français sur la réalisation de trachéotomies et les soins de trachéotomies pendant la pandémie de COVID-19

text: ABSTRACT Tracheostomy post-tracheostomy care are regarded as at high risk for contamination of health care professionals with the new coronavirus (SARS-CoV-2). Considering the rapid spread of the infection, all patients in France must be considered as potentially infected by the virus. Nevertheless, patients without clinical or radiological (CT scan) markers of COVID-19, and with negative nasopharyngeal sample within 24h of surgery, are at low risk of being infected. Instructions for personal protection include specific wound dressings and decontamination of all material used. The operating room should be ventilated after each tracheostomy and the pressure of the room should be neutral or negative. Percutaneous tracheostomy is to be preferred over surgical cervicotomy in order to reduce aerosolization and to avoid moving patients from the intensive care unit to the operating room. Ventilation must be optimized during the procedure, to limit patient oxygen desaturation. Drug assisted neuromuscular blockage is advised to reduce coughing during tracheostomy tube insertion. An experienced team is mandatory to secure and accelerate the procedure as well as to reduce risk of contamination.


title: Can SARS-CoV-2-infected women breastfeed after viral clearance?

text: 浙江大学医学院附属第一医院感染科收治一名新型冠状病毒 (SARS-CoV-2) 感染的妊娠晚期孕妇. 在其住院期间, 患者进行了剖腹产术, 新生儿被证实无 SARS-CoV-2 感染. 结合已有的相关文献报道, SARS-CoV-2 通过母婴垂直传播的依据不足, 同时乳汁的病毒检测阴性, 因此目前不能证明 SARS-CoV-2 病毒能通过乳汁传播. 通过研究, 我们建议感染 SARS-CoV-2 的孕妇, 在治愈出院后, 应经过两周隔离观察, 并且病毒复查结果为阴性, 才可以进行母乳喂养.


title: Alert to a European epidemic

text: Funds must be forthcoming for an effective EU Centre for Disease Control.


title: Identification, Expression, and Processing of an 87-kDa Polypeptide Encoded by ORF 1a of the Coronavirus Infectious Bronchitis Virus

text: Abstract Nucleotide sequence analysis has shown previously that the genomic-length mRNA (mRNA1) of the coronavirus infectious bronchitis virus (IBV) contains two large open reading frames (ORFs), 1a and 1b, with the potential to encode polyproteins of approximately 441 and 300 kDa, respectively. We have characterized the specificity of a set of region-specific antisera raised against the 5′-portion of ORF 1a by immunoprecipitation of in vitro-synthesized, C-terminally truncated 1a polypeptides and used these antisera to detect virus-specific proteins in IBV-infected Vero cells. Two antisera, which had specificity for IBV sequences from nucleotides 710 to 2079 and 1355 to 2433, respectively, immunoprecipitated a polypeptide of approximately 87 kDa from IBV-infected Veto cells. In vitro translation of ORF 1a sequence terminating at nucleotide 5763 did not produce this protein unless the in vitro translation products were incubated with Vero cell S10 extracts prepared from either IBV-infected or mock-infected Vero cells. However, processing of the 87-kDa protein was also observed when the same region was expressed in Vero cells using the vaccinia virus/T7 expression system. This observation indicates that the 87-kDa polypeptide is encoded within the 5′-most 3000 nucleotides of mRNA 1 and that it might be cleaved from the 1a polyprotein by viral and cellular proteinases.


title: Frailty in Orthopaedics: Is Age Relevant?

text: Over the last decade, national guidelines and the Best Practice Tariff (BPT) have been created to incentivise quality care in patients aged over 60 with hip fractures. This has resulted in significantly decreased length of stay, mortality and post-operative complications in this patient cohort. However, there is increasing recognition of frail patients in all age groups sustaining all fragility fractures. Until recently, these patients experienced poorer outcomes and were excluded from the dedicated care pathways that hip fracture patients received. The BPT and other national guidelines are now expanding inclusion criteria into care packages between guidelines which were initially reserved for hip fracture patients. This expansion is placing increasing pressure on limited NHS resources. Current variations between society guidelines risks producing regional and departmental inconsistencies in care. There is therefore a need to provide consistent guideline targeted at the most vulnerable trauma patients of this expanded cohort. Although the current BPT applies to over 60s only, there is limited evidence to support age-related prognosis in trauma. In contrast, frailty is being increasingly recognised as a global indicator of patient outcomes irrespective of age, with use of Clinical Frailty Scale (CFS) being adopted in various medical fields. BOAST is already using CFS as an inclusion criterion for major trauma and there is increasing data to suggest that frail trauma patients benefit most from comprehensive geriatric care and expedient time-to-operation. We suggest that CFS should take precedence over age when ascertaining clinical priority and producing Best Practice Tariffs. Further research is required to investigate frailty-related outcomes in trauma and the impact of comprehensive care bundles on the outcomes of frail orthopaedic patients.


title: Hypothalamus-pituitary-adrenal axis involves in anti-viral ability through regulation of immune response in piglets infected by highly pathogenic porcine reproductive and respiratory syndrome virus

text: BACKGROUND: The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) has been responsible for several viral attacks in the Asian porcine industry, since the first outbreak in China in 2006. During the early stages of the HP-PRRSV infection, high levels of proinflammatory cytokines are noted in the host peripheral blood, which are accompanied by severe lesions in the lungs and immune system organs; these are considered as the greatest contributors to the overall disease burden. We hypothesized that the anti-PRRSV response in piglets might be mediated by the hypothalamus-pituitary-adrenal (HPA) axis, which led to a decrease in the psycho-neuroendocrinological manifestation of HP-PRRSV etiology via immune response regulation. RESULTS: We investigated the regulation of the HPA axis in HP-PRRSV-infected piglets that were treated with 1 mg/kg body weight (b. w.)/day mifepristone (RU486) or 2 mg/kg b.w./day dexamethasone (DEX). Both RU486 and DEX enhanced the disease status of the piglets infected by the HP-PRRSV HuN4 strain, resulting in high mortality and more severe pathological changes in the lungs. CONCLUSIONS: HP-PRRSV infection activates the HPA axis, and artificial regulation of the immune-endocrine system enhances disease severity in HP-PRRSV-infected piglets. Thus, DEX and RU486 should be avoided in the clinical treatment of HP-PRRS. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1414-3) contains supplementary material, which is available to authorized users.


title: SARS: Systematic Review of Treatment Effects

text: BACKGROUND: The SARS outbreak of 2002–2003 presented clinicians with a new, life-threatening disease for which they had no experience in treating and no research on the effectiveness of treatment options. The World Health Organization (WHO) expert panel on SARS treatment requested a systematic review and comprehensive summary of treatments used for SARS-infected patients in order to guide future treatment and identify priorities for research. METHODS AND FINDINGS: In response to the WHO request we conducted a systematic review of the published literature on ribavirin, corticosteroids, lopinavir and ritonavir (LPV/r), type I interferon (IFN), intravenous immunoglobulin (IVIG), and SARS convalescent plasma from both in vitro studies and in SARS patients. We also searched for clinical trial evidence of treatment for acute respiratory distress syndrome. Sources of data were the literature databases MEDLINE, EMBASE, BIOSIS, and the Cochrane Central Register of Controlled Trials (CENTRAL) up to February 2005. Data from publications were extracted and evidence within studies was classified using predefined criteria. In total, 54 SARS treatment studies, 15 in vitro studies, and three acute respiratory distress syndrome studies met our inclusion criteria. Within in vitro studies, ribavirin, lopinavir, and type I IFN showed inhibition of SARS-CoV in tissue culture. In SARS-infected patient reports on ribavirin, 26 studies were classified as inconclusive, and four showed possible harm. Seven studies of convalescent plasma or IVIG, three of IFN type I, and two of LPV/r were inconclusive. In 29 studies of steroid use, 25 were inconclusive and four were classified as causing possible harm. CONCLUSIONS: Despite an extensive literature reporting on SARS treatments, it was not possible to determine whether treatments benefited patients during the SARS outbreak. Some may have been harmful. Clinical trials should be designed to validate a standard protocol for dosage and timing, and to accrue data in real time during future outbreaks to monitor specific adverse effects and help inform treatment.


title: A Proof Of Concept For A Syndromic Surveillance System Based On Routine Ambulance Records In The South-west Of England, For The Influenza Season 2016/17

text: The introduction of electronic patient records in the ambulance service provides new opportunities to monitor the population. Most patients presenting to British ambulance services are discharged at scene. Ambulance records are therefore an ideal data source for syndromic early event detection systems to monitor infectious disease in the prehospital population. It has been previously found that tympanic temperature records can be used to detect influenza outbreaks in emergency departments. This study investigated whether routine tympanic temperature readings collected by ambulance crews can be used to detect seasonal influenza. Here we show that these temperature readings do allow the detection of seasonal influenza before methods applied to conventional data sources. The counts of pyretic patients were used to calculate a sliding case ratio (CR) as a measurement to detect seasonal influenza outbreaks. This method does not rely on conventional thresholds and can be adapted to the data. The data collected correlated with seasonal influenza. The 2016/17 outbreak was detected with high specificity and sensitivity, up to 9 weeks before other surveillance programs. An unanticipated outbreak of E. coli was detected in the same dataset. Our results show that ambulance records can be a useful data source for biosurveillance systems. Two outbreaks caused by different infectious agents have been successfully detected. The routine ambulance records allowed to use tympanic temperature readings that can be used as surveillance tool for febrile diseases. Therefore, this method is a valuable addition to the current surveillance tools.


title: Addiction telemedicine comes into its own with COVID‐19

text: Telepsychiatry, telemental health, telepsychology, teletherapy — whatever you call it, it is now required in the era of the COVID‐19 pandemic. “We should be doing it immediately; the more, the better,” said Robert Caudill, M.D., a member of the American Psychiatric Association's Committee on Telepsychiatry. “It's exploded in the past week, obviously,” he told ADAW in a Zoom interview on March 23. But he thinks it should be the norm.


title: Population-scale Longitudinal Mapping of COVID-19 Symptoms, Behavior, and Testing Identifies Contributors to Continued Disease Spread in the United States

text: Despite social distancing and shelter-in-place policies, COVID-19 continues to spread in the United States. A lack of timely information about factors influencing COVID-19 spread and testing has hampered agile responses to the pandemic. We developed How We Feel, an extensible web and mobile application that aggregates self-reported survey responses, to fill gaps in the collection of COVID-19-related data. How We Feel collects longitudinal and geographically localized information on users' health, behavior, and demographics. Here we report results from over 500,000 users in the United States from April 2, 2020 to May 12, 2020. We show that self- reported surveys can be used to build predictive models of COVID-19 test results, which may aid in identification of likely COVID-19 positive individuals. We find evidence among our users for asymptomatic or presymptomatic presentation, as well as for household and community exposure, occupation, and demographics being strong risk factors for COVID-19. We further reveal factors for which users have been SARS-CoV-2 PCR tested, as well as the temporal dynamics of self- reported symptoms and self-isolation behavior in positive and negative users. These results highlight the utility of collecting a diverse set of symptomatic, demographic, and behavioral self- reported data to fight the COVID-19 pandemic.


Canada 2010: what should global health expect?


title: Novel activities of safe-in-human broad-spectrum antiviral agents

text: Abstract According to the WHO, there is an urgent need for better control of viral diseases. Re-positioning existing safe-in-human antiviral agents from one viral disease to another could play a pivotal role in this process. Here, we reviewed all approved, investigational and experimental antiviral agents, which are safe in man, and identified 59 compounds that target at least three viral diseases. We tested 55 of these compounds against eight different RNA and DNA viruses. We found novel activities for dalbavancin against echovirus 1, ezetimibe against human immunodeficiency virus 1 and Zika virus, as well as azacitidine, cyclosporine, minocycline, oritavancin and ritonavir against Rift valley fever virus. Thus, the spectrum of antiviral activities of existing antiviral agents could be expanded towards other viral diseases.


title: Cold plasma, a new hope in the field of virus inactivation

text: Abstract Viruses can infect all cell-based organisms, from bacteria to humans, animals, and plants. They are responsible for numerous cases of hospitalization, many deaths, and widespread crop destruction, which all result in an enormous medical, economical, and biological burden. Each of the currently used decontamination methods have important drawbacks. Cold plasma has entered this field as a novel, efficient, and clean solution for virus inactivation. Here, we present the recent developments in this promising field of cold-plasma-mediated virus inactivation, and describe the applications and mechanisms of the inactivation. This is a particularly relevant subject as viral pandemics, such as the COVID-19 pandemic, expose the need for alternative viral inactivation methods to replace, complement or upgrade existing ones.


title: Chapter 40 Infectious Demyelinating Diseases

text: Publisher Summary This chapter focuses on three human CNS demyelinating diseases—namely, acute disseminated encephalomyelitis (ADEM), also known as postinfectious encephalomyelitis, progressive multifocal leukoencephalopathy (PML), and multiple sclerosis (MS). These three diseases have very different clinical courses and distinctive pathological features, although all share the essential element of demyelination. ADEM and PML have antithetic modes of pathogenesis. ADEM is a predominantly extraneural infection resulting in a virus-induced host autoimmune response, whereas PML is a direct lytic infection of oligodendrocytes in an immunocompromised host. It is clear that a diverse array of viruses can infect the human central nervous system. The viral infections can result in a wide variety of clinical and pathological symptoms. While some viruses may cause widespread inflammation and neurodegeneration, other viruses may remain latent in the CNS and only produce pathological changes during reactivation. Viral induced demyelination can occur both from direct infection of the myelin-producing oligodendrocytes, as in PML, or by indirect mechanisms that have yet to be determined. The outcome of a viral CNS infection depends not only on viral characteristics, but also on the interaction between virus and host cell.


title: The contagion of mortality: A terror management health model for pandemics

text: The novel coronavirus, COVID‐19, proliferates as a contagious psychological threat just like the physical disease itself. Due to the growing death toll and constant coverage this pandemic gets, it is likely to activate mortality awareness, to greater or lesser extents, depending on a variety of situational factors. Using terror management theory and the terror management health model, we outline reactions to the pandemic that consist of proximal defences aimed at reducing perceived vulnerability to (as well as denial of) the threat, and distal defences bound by ideological frameworks from which symbolic meaning can be derived. We provide predictions and recommendations for shifting reactions to this pandemic towards behaviours that decrease, rather than increase, the spread of the virus. We conclude by considering the benefits of shifting towards collective mindsets to more effectively combat COVID‐19 and to better prepare for the next inevitable pandemic.


title: Cardiac function in relation to myocardial injury in hospitalised patients with COVID-19

text: BACKGROUND: Previous studies have reported on myocardial injury in patients with coronavirus infectious disease 19 (COVID-19) defined as elevated cardiac biomarkers. Whether elevated biomarkers truly represent myocardial dysfunction is not known. The aim of this study was to explore the incidence of ventricular dysfunction and assess its relationship with biomarker analyses. METHODS: This cross-sectional study ran from April 1 to May 12, 2020, and consisted of all consecutively admitted patients to the Radboud university medical centre nursing ward for COVID-19. Laboratory assessment included high-sensitivity Troponin T and N‑terminal pro-B-type natriuretic peptide (NT-proBNP). Echocardiographic evaluation focused on left and right ventricular systolic function and global longitudinal strain (GLS). RESULTS: In total, 51 patients were included, with a median age of 63 years (range 51–68 years) of whom 80% was male. Troponin T was elevated (>14 ng/l) in 47%, and a clinically relevant Troponin T elevation (10 × URL) was found in three patients (6%). NT-proBNP was elevated (>300 pg/ml) in 24 patients (47%), and in four (8%) the NT-proBNP concentration was >1,000 pg/ml. Left ventricular dysfunction (ejection fraction <52% and/or GLS >−18%) was observed in 27%, while right ventricular dysfunction (TAPSE <17 mm and/or RV S’ < 10 cm/s) was seen in 10%. There was no association between elevated Troponin T or NT-proBNP and left or right ventricular dysfunction. Patients with confirmed pulmonary embolism had normal right ventricular function. CONCLUSIONS: In hospitalised patients, it seems that COVID-19 predominantly affects the respiratory system, while cardiac dysfunction occurs less often. Based on a single echocardiographic evaluation, we found no relation between elevated Troponin T or NT-proBNP, and ventricular dysfunction. Echocardiography has limited value in screening for ventricular dysfunction. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12471-020-01458-2) contains supplementary material, which is available to authorized users.


title: Phylodynamic applications in 21(st) century global infectious disease research

text: BACKGROUND: Phylodynamics, the study of the interaction between epidemiological and pathogen evolutionary processes within and among populations, was originally defined in the context of rapidly evolving viruses and used to characterize transmission dynamics. The concept of phylodynamics has evolved since the early 21(st) century, extending its reach to slower-evolving pathogens, including bacteria and fungi, and to the identification of influential factors in disease spread and pathogen population dynamics. RESULTS: The phylodynamic approach has now become a fundamental building block for the development of comparative phylogenetic tools capable of incorporating epidemiological surveillance data with molecular sequences into a single statistical framework. These innovative tools have greatly enhanced scientific investigations of the temporal and geographical origins, evolutionary history, and ecological risk factors associated with the growth and spread of viruses such as human immunodeficiency virus (HIV), Zika, and dengue and bacteria such as Methicillin-resistant Staphylococcus aureus. CONCLUSIONS: Capitalizing on an extensive review of the literature, we discuss the evolution of the field of infectious disease epidemiology and recent accomplishments, highlighting the advancements in phylodynamics, as well as the challenges and limitations currently facing researchers studying emerging pathogen epidemics across the globe. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s41256-017-0034-y) contains supplementary material, which is available to authorized users.


title: Diabetes care during COVID-19 lockdown at a tertiary care centre in India

text: Abstract Providing scheduled consultations to persons with diabetes during the COVID-19 induced lockdowns posed a major challenge. With the clinicians occupied in COVID management, a strategy of using telemedicine and engaging a team of para-clinical doctors was devised. Telephonic follow up consults were given and diabetes care was efficiently delivered.


title: Antiviral activity of Alpinia katsumadai extracts against rotaviruses

text: Abstract In vitro anti-rotavirus activity of Alpinia katsumadai (AK) extracts were evaluated against bovine G8P[7] and porcine G5P[7] rotaviruses in two different assay strategies, a mixed treatment assay and a post treatment assay. In the mixed treatment assay, six AK extracts [AK-1 (EtOH extract), AK-3 (H2O layer), AK-5 (40% methanol fraction), and AK-9–11 (H2O extract, polysaccharide fraction, supernatant fraction)] exhibited inhibitory activities against G5P[7] rotavirus with the EC50 values ranging from 0.7±0.4 to 33.7±6.5μg/mL. Extracts AK-1, AK-3, and AK-5 inhibited rotavirus infection against G8P[7] rotavirus, the with EC50 values of 8.4±2.2μg/mL, 6.5±0.8μg/mL and 8.4±5.0μg/mL, respectively. By hemagglutination inhibition (HI) assay, six AK extracts completely inhibited viral adsorption onto human RBCs in both strains of rotaviruses at less than 11μg/mL. However, in the post treatment assay, there was no anti activity shown against both strains of rotaviruses. As a result, six AK extracts were attributed mainly to having a strong interaction with hemagglutinin protein on the outer surface of rotavirus, resulting to blockage of viral adsorption.


title: Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera

text: Severe fever with thrombocytopenia syndrome (SFTS) caused by the SFTS virus (SFTSV), a phlebovirus in the family Bunyaviridae, is an emerging tick-borne infectious disease that impacts humans. This disease manifests as a decreased blood cell count and multi-organ failure, with a case-fatality rate of more than 12% in China. Because vaccines or antiviral drugs for the treatment of this disease are not available, monitoring the SFTS circulation in animals and controlling the tick-mammal cycle are important for preventing SFTS. Monoclonal antibodies against the recombinant nucleoprotein of SFTSV were generated to develop a competitive enzyme-linked immunosorbent assay (cELISA) for the detection of antibodies against SFTSV infection in cattle. The specificity and sensitivity of cELISA was assessed by comparing the results of this assay to those of an immunofluorescence assay (IFA). The results of the cELISA using 416 field bovine serum samples and laboratory-immunized positive sera showed 98.1% consistency with those of the IFA. The cELISA used in this study did not show cross-reactivity with antisera against other viral cattle diseases. The cELISA presented in this study can be applied to detect antibodies against SFTSV in cattle.


title: Different pattern of viral infections and clinical outcomes in patient with acute exacerbation of chronic obstructive pulmonary disease and chronic obstructive pulmonary disease with pneumonia

text: Respiratory viruses are well‐known causes of acute exacerbation of chronic obstructive pulmonary disease (AE‐COPD) and also important pathogens for concomitant pneumonia in COPD (CP‐COPD). However, the differences in a viral infection pattern and clinical impacts of respiratory viruses between the two groups have not been well investigated. The clinical and microbiological data from COPD patients admitted with AE‐COPD (n = 281) or CP‐COPD (n = 284) between January 2010 and December 2012 were reviewed. After excluding 88 patients (40 with AE‐COPD and 48 with CP‐COPD) who did not undergo a multiplex RT‐PCR test for respiratory viruses, the demographic characteristics, identified viruses, and clinical outcomes of the AE‐COPD and CP‐COPD groups were compared. Respiratory viruses were identified in 41.9% of AE‐COPD group and 33.5% of the CP‐COPD groups. The most common virus was influenza virus in the AE‐COPD group (33.7%) versus human coronavirus (24.1%) in the CP‐COPD group. Influenza virus was significantly more common in the AE‐ACOPD group than in the CP‐COPD group (P < 0.01). In‐hospital mortality of AE‐COPD and CP‐COPD were 1.2% and 12.3%, respectively (P < 0.01). Among CP‐COPD patients, in‐hospital mortality of patients with only viral infection group, only bacterial infection group, and viral‐bacterial co‐infection were 2.6%, 25.8%, and 17.5%, respectively (P = 0.01). Respiratory viruses were commonly identified in both AE‐COPD and CP‐COPD, influenza virus and human coronavirus were the most common viruses identified in AE‐COPD and CP‐COPD patients, respectively. The mortality rates of only viral infection group was significantly lower than only bacterial infection or viral‐bacterial co‐infection group in CP‐COPD patients. J. Med. Virol. 88:2092–2099, 2016. © 2016 Wiley Periodicals, Inc.


title: The Se S/N interactions as a possible mechanism of δ-aminolevulinic acid dehydratase enzyme inhibition by organoselenium compounds: a computational study

text: Organoselenium compounds present many pharmacological properties and are promising drugs. However, toxicological effects associated with inhibition of thiol-containing enzymes, such as the δ-aminolevulinic acid dehydratase (δ-AlaD), have been described. The molecular mechanism(s) by which they inhibit thiol-containing enzymes at the atomic level, is still not well known. The use of computational methods to understand the physical-chemical properties and biological activity of chemicals is essential to the rational design of new drugs. In this work, we propose an in silico study to understand the δ-AlaD inhibition mechanism by diphenyl diselenide (DPDS) and its putative metabolite, phenylseleninic acid (PSA), using δ-AlaD enzymes from Homo sapiens (Hsδ-AlaD), Drosophila melanogaster (Dmδ-AlaD) and Cucumis sativus (Csδ-AlaD). Protein modeling homology, molecular docking, and DFT calculations are combined in this study. According to the molecular docking, DPDS and PSA might bind in the Hsδ-AlaD and Dmδ-AlaD active sites interacting with the cysteine residues by Se(…)S interactions. On the other hand, the DPDS does not access the active site of the Csδ-AlaD (a non-thiol protein), while the PSA interacts with the amino acids residues from the active site, such as the Lys291. These interactions might lead to the formation of a covalent bond, and consequently, to the enzyme inhibition. In fact, DFT calculations (mPW1PW91/def2TZVP) demonstrated that the selenylamide bond formation is energetically favored. The in silico data showed here are in accordance with previous experimental studies, and help us to understand the reactivity and biological activity of organoselenium compounds.


How to face the novel coronavirus infection during the 2019–2020 epidemic: the experience of Sichuan Provincial People’s Hospital


title: A novel immunochromatographic system for easy-to-use detection of group 1 avian influenza viruses with acquired human-type receptor binding specificity

text: Abstract A switch of viral hemagglutinin receptor binding specificity from bird-type α2,3- to human-type α2,6-linked sialic acid is necessary for an avian influenza virus to become a pandemic virus. In this study, an easy-to-use strip test to detect receptor binding specificity of influenza virus was developed. A biotinylated anti-hemagglutinin antibody that bound a broad range of group 1 influenza A viruses and latex-conjugated α2,3 (blue) and α2,6 (red) sialylglycopolymers were used in an immunochromatographic strip test, with avidin and lectin immobilized on a nitrocellulose membrane at test and control lines, respectively. Accumulation of a sialylglycopolymer–virus–antibody complex at the test line was visualized by eye. The strip test could be completed in 30min and did not require special equipment or skills, thereby avoiding some disadvantages of current methods for analyzing receptor binding specificity of influenza virus. The strip test could detect the receptor binding specificity of a wide range of influenza viruses, as well as small increases in the binding affinity of variant H5N1 viruses to α2,6 sialylglycans at viral titers >128 hemagglutination units. The strip test results were in agreement with those of ELISA virus binding assays, with correlations >0.95. In conclusion, the immunochromatographic strip test developed in this study should be useful for monitoring potential changes in the receptor binding specificity of group 1 influenza A viruses in the field.


Serum amyloid A is a predictor for prognosis of COVID‐19


title: Using dimension reduction to improve outbreak predictability of multistrain diseases

text: Multistrain diseases have multiple distinct coexisting serotypes (strains). For some diseases, such as dengue fever, the serotypes interact by antibody-dependent enhancement (ADE), in which infection with a single serotype is asymptomatic, but contact with a second serotype leads to higher viral load and greater infectivity. We present and analyze a dynamic compartmental model for multiple serotypes exhibiting ADE. Using center manifold techniques, we show how the dynamics rapidly collapses to a lower dimensional system. Using the constructed reduced model, we can explain previously observed synchrony between certain classes of primary and secondary infectives (Schwartz et al. in Phys Rev E 72:066201, 2005). Additionally, we show numerically that the center manifold equations apply even to noisy systems. Both deterministic and stochastic versions of the model enable prediction of asymptomatic individuals that are difficult to track during an epidemic. We also show how this technique may be applicable to other multistrain disease models, such as those with cross-immunity.


title: Pandemic influenza A(H1N1) virus infection in solid organ transplant recipients: impact of viral and non-viral co-infection

text: Solid organ transplant recipients (SOTR) are at risk of serious influenza-related complications. The impact of respiratory co-infection in SOTR with 2009 pandemic influenza A(H1N1) is unknown. A multicentre prospective study of consecutive cases of pandemic influenza A(H1N1) in SOTR was carried out to assess the clinical characteristics and outcome and the risk factors for co-infection. Overall, 51 patients were included. Median time from transplant was 3.7 years, 5.9% of the cases occurred perioperatively and 7.8% were hospitalacquired. Pneumonia was diagnosed in 15 (29.4%) patients. Ten cases were severe (19.6%): 13.7% were admitted to intensive care units, 5.9% suffered septic shock, 5.9% developed acute graft rejection and 7.8% died. Co-infection was detected in 15 patients (29.4%): eight viral, six bacterial and one fungal. Viral co-infection did not affect the outcome. Patients with non-viral co-infection had a worse outcome: longer hospital stay (26.2 ± 20.7 vs. 5.5 ± 10.2) and higher rate of severe diseases (85.7% vs. 2.3%) and mortality (42.8% vs. 2.3%). Independent risk factors for non-viral co-infection were: diabetes mellitus and septic shock. Other factors associated with severe influenza were: delayed antiviral therapy, diabetes mellitus, time since transplantation <90 days and pneumonia. In conclusion, pandemic influenza A can cause significant direct and indirect effects in SOTR, especially in the early post-transplant period, and should be treated early. Clinicians should be aware of the possibility of non-viral co-infection, mainly in diabetic patients and severe cases. An effort should be made to prevent influenza with immunization of the patient and the environment.


title: Understanding the public temper through an evaluation of rumours: an ethnographical method using educational technology

text: The power of rumours is that they can be broadly exchanged, generating a ‘public temper’ (which is everybody’s temper without being anybody’s temper in particular). This article, therefore, describes an approach to measuring the public temper, examining particularly the public temper of an Arab society, namely Saudi Arabia. It addresses the following research question: is it possible to analyse existing (scholarly) rumours to see if they can be used as informants of the public temper of the culture in which they exist? This question is answered ethnographically by analysing 579 Arabic online rumours collected by students as part of their critical engagement with educational technology. Having analysed the data, four categories emerged: the concerns, interests, attitudes and values of Saudi Arabia. According to the literature, these four categories, taken together, constitute the emotional domain (i.e., the public temper) of a society. Thus, a theoretical proposition (and contribution to the existing literature regarding sociology) is that rumours mirror the public temper of a culture, reflecting a range of emotions from simple to complex (from concerns, interests and attitudes to values). Simpler emotions (e.g., concerns) appear to be more easily affected by rumours than more complex emotions (e.g., values). An implication of this study is that rumours have ‘biographies’, which detail public tempers across space and time. Rumours are ‘records’ of public tempers that should be read in the same way archaeologists read landscapes and remains. Although rumours entail ill-defined information, it is feasible to well define society through such ill-defined information, meaning that something can come out of its opposite. This study offers ethnographers a new method of understanding public tempers through rumours, alongside conventional meaning-making symbols (e.g., poems).


title: Identification of 9-O acetyl sialoglycoconjugates (9-OAcSGs) as biomarkers in childhood acute lymphoblastic leukemia using a lectin, Achatinin(H), as a probe

text: Neoplastic transformation causes changes in cell surface architecture, most notably, aberrant sialylation. Exploiting the restricted specificity of a 9-O acetyl sialic acid (9-OAcSA) binding lectin, Achatinin(H) (ATN(H)), we have identified two 9-O acetyl sialoglyconjugates (9-OAcSGs) on lymphoblasts of 87 children suffering from acute lymphoblastic leukemia (ALL). The preferential binding of ATN(H) to lymphoblasts induces their 11-fold increased agglutination (81 ± 7.8%) compared to peripheral blood mononuclear cells (PBMC) of normal donors (8 ± 4.3%) which corroborates with flow cytometry studies. Agglutination of MOLT-4 (87 ± 4.8%), a lymphoblastoid cell line and MDCK (91.25 ± 0.01%), a cell line expressing surface 9-OAcSA, confirms the preferential binding of ATN(H) to lymphoblasts through their surface 9-OAcSGs. Furthermore, fluorometric quantitation reveals a 4.6-fold increase in % of 9-OAcSA on lymphoblasts of ALL patients (42.1 ± 4.1%) compared to normal donors (9.2. ± 3.4%). Western blotting confirms that ATN(H) recognizes two membrane sialoglycoconjugates, of MW 120 kDa and 90 kDa, both having 9-OAcSA α2 → 6 GalNAc terminal sugar moiety as their lectinogenic epitope. We propose that these 9-OAcSGs may serve as biomarkers for detection and monitoring of lymphoblasts in ALL and accordingly merit therapeutic considerations.


title: Review on heterogeneous photocatalytic disinfection of waterborne, airborne, and foodborne viruses: Can we win against pathogenic viruses?

text: Microbial pathogenic contaminations have world widely represented a serious health hazard to humans. Viruses, as a member of microbial contaminants, seriously threaten human health due to their high environmental resistance, having small sizes, and causing an extensive range of diseases. Therefore, selecting an appropriate technology to remove viral contaminants from the air, water, and foods is of prominent significance. Traditional methods for viral disinfection have not proven to be highly practical and effective because they need high energy resources and operational expenses. In recent years, semiconductor-based photocatalysis has attracted more attention in the field of microorganism inactivation due to its outstanding performance and mild reaction conditions. Therefore, this review primarily concentrates on the recent development in viral inactivation/disinfection by heterogeneous photocatalysts. Moreover, the photocatalytic viral inactivation of waterborne, airborne, and foodborne viruses is discussed. Given the appealing merits of heterogeneous photocatalytic disinfection of viruses, there is no doubt that this technology will be an impressively active research field and a source of comfort and confidence to humans in battling against viruses.


title: Influenza virus-related critical illness: pathophysiology and epidemiology

text: Influenza virus affects the respiratory tract by direct viral infection or by damage from the immune system response. In humans, the respiratory epithelium is the only site where the hemagglutinin (HA) molecule is effectively cleaved, generating infectious virus particles. Virus transmission occurs through a susceptible individual’s contact with aerosols or respiratory fomites from an infected individual. The inability of the lung to perform its primary function of gas exchange can result from multiple mechanisms, including obstruction of the airways, loss of alveolar structure, loss of lung epithelial integrity from direct epithelial cell killing, and degradation of the critical extracellular matrix. Approximately 30–40% of hospitalized patients with laboratory-confirmed influenza are diagnosed with acute pneumonia. These patients who develop pneumonia are more likely to be < 5 years old, > 65 years old, Caucasian, and nursing home residents; have chronic lung or heart disease and history of smoking, and are immunocompromised. Influenza can primarily cause severe pneumonia, but it can also present in conjunction with or be followed by a secondary bacterial infection, most commonly by Staphylococcus aureus and Streptococcus pneumoniae. Influenza is associated with a high predisposition to bacterial sepsis and ARDS. Viral infections presenting concurrently with bacterial pneumonia are now known to occur with a frequency of 30–50% in both adult and pediatric populations. The H3N2 subtype has been associated with unprecedented high levels of intensive care unit (ICU) admission. Influenza A is the predominant viral etiology of acute respiratory distress syndrome (ARDS) in adults. Risk factors independently associated with ARDS are age between 36 and 55 years old, pregnancy, and obesity, while protective factors are female sex, influenza vaccination, and infections with Influenza A (H3N2) or Influenza B viruses. In the ICU, particularly during the winter season, influenza should be suspected not only in patients with typical symptoms and epidemiology, but also in patients with severe pneumonia, ARDS, sepsis with or without bacterial co-infection, as well as in patients with encephalitis, myocarditis, and rhabdomyolysis.


title: What is the importance of zoonotic trichomonads for human health?

text: Trichomonads are common parasites of many vertebrate and invertebrate species, with four species classically recognized as human parasites: Dientamoeba fragilis, Pentatrichomonas hominis, Trichomonas vaginalis, and Trichomonas tenax. The latter two species are considered human-specific; by contrast, D. fragilis and P. hominis have been isolated from domestic and farm mammals, demonstrating a wide host range and potential zoonotic origin. Several new studies have highlighted the zoonotic dimension of trichomonads. First, species typically known to infect birds and domestic mammals have been identified in human clinical samples. Second, several phylogenetic analyses have identified animal-derived trichomonads as close sister taxa of the two human-specific species. It is our opinion, therefore, that these observations prompt further investigation into the importance of zoonotic trichomonads for human health.


The clinical need for the RVP test


title: Nucleic Acids-Based Therapeutics in the Battle Against Pathogenic Viruses

text: For almost three decades, researchers have studied the possibility to use nucleic acids as antiviral therapeutics. In theory, compounds such as antisense oligonucleotides, ribozymes, DNAzymes, and aptamers can be designed to trigger the sequence-specific inhibition of particular mRNA transcripts, including viral genomes. However, difficulties with their efficiency, off-target effects, toxicity, delivery, and stability halted the development of nucleic acid-based therapeutics that can be used in the clinic. So far, only a single antisense drug, Vitravene for the treatment of CMV-induced retinitis in AIDS patients, has made it to the clinic. Since the discovery of RNA interference (RNAi), there is a renewed interest in the development of nucleic acid-based therapeutics. Antiviral RNAi approaches are highly effective in vitro and in animal models and are currently being tested in clinical trials. Here we give an overview of antiviral nucleic acid-based therapeutics. We focus on antisense and RNAi-based compounds that have been shown to be effective in animal model systems.


title: It's not about the money. EU funds, local opportunities, and Euroscepticism

text: Growing Euroscepticism across the European Union (EU) leaves open questions as to what citizens expect to gain from EU Membership and what influences their dissent for EU integration. This paper looks at the EU Structural Funds, one of the largest and most visible expenditure items in the EU budget, to test their impact on electoral support for the EU. By leveraging the Referendum on Brexit held in the United Kingdom, a spatial RDD analysis offers causal evidence that EU money does not influence citizens' support for the EU. Conversely, the analysis shows that EU funds mitigate Euroscepticism only where they are coupled by tangible improvements in local labour market conditions, the ultimate objective of this form of EU intervention. Money cannot buy love for the EU, but its capacity to generate new local opportunities certainly can.


title: Mimicry of human IgE epitopes by anti-idiotypic antibodies 1 1 Edited by I. A. Wilson

text: Abstract According to Jerne’s network hypothesis, the binding site of an anti-idiotypic antibody also represents the internal image of an epitope present on a foreign, or even a self antigen. In recent years, antigen mimicry has been defined at the molecular level for some xeno-antigens. However, until now there has been no demonstration of structural mimicry between a human anti-idiotypic antibody and a self structure. To address this question, we used human IgE as the self structure and a well-defined anti-human IgE mAb (BSW17). We describe the isolation of two anti- idiotypic antibodies specific for the anti-IgE antibody BSW17 from a non-immune human Fab phage display library. Interestingly, these two anti-idiotypic antibodies mimic the same molecular surface region as a previously described IgE peptide mimotope isolated by panning on BSW17, but they cover a much larger epitope on the IgE molecule. Accordingly, immunisation of rabbits with the two anti-idiotypic antibodies induced high-affinity antibodies with the same characteristics as BSW17. Thus, our data demonstrate that it is possible to isolate anti-idiotypic antibodies derived from the human genome without the need for hyperimmunisation, and confirm Jerne’s hypothesis that both foreign antigens and self structures can be mimicked by our own immunoglobulins.


title: BWR develops mobile fuel cell generator sanitation systems

text: Blue Water Resolute (BWR) Innovations in North Dakota, USA has developed a line of small, mobile fuel cell generator sanitation systems, featuring a 10 kW electrical generator on a movable cart. The SFC 110 Fuel Cell Thermal Sanitizer combines heat produced by a fuel cell with ultraviolet light to inactivate a full spectrum of biological contaminants – including viruses, bacteria and fungi – while providing uniform sanitisation coverage with minimal risk of material compatibility issues.


title: The construction of a hospital disease tracking and control system with a disease infection probability model

text: With relatively short latency and rapid propagation, viral diseases could be transmitted through the air to medical personnel or the public during the incubation period. To reduce the possibilities of spread, this research creates an infection probability model based on the settling velocity and concentration distribution of infectious droplets. Then, radio frequency identification (RFID) technology is employed to track the travel history (time, date and place) of the infected patients. A tree structure algorithm and an infection probability model are applied to trace the transmission routes, discover the correlations between carriers and suspected cases, and finally calculate the infection probability on the basis of time interval. In case of an epidemic outbreak or once an infected case is confirmed, the disease tracking and control system could be initiated by accessing RFID logs to plot the carriers’ time of onset and to trace possible routes of transmission via tree diagrams. The disease tracking and control system developed in this research can assist hospitals in assessing the risk of infection among medical personnel, as well as in prompt implementation of infection prevention and control measures, in order to reduce hospital acquired infections and provide a safe health care setting.


Liver and Kidney Injuries in COVID-19 and Their Effects on Drug Therapy; a Letter to Editor


title: The occurrence of polyomaviruses WUPyV and KIPyV among patients with severe respiratory infections

text: In 2007, the new polyomaviruses WUPyV and KIPyV were identified in patients with acute respiratory infections. The aim of this study was to investigate these viruses in hospitalized patients with severe acute respiratory infection (SARI). A retrospective study was conducted with 251 patients, from April 2009 to November 2010, using nasopharyngeal aspirates, naso- and oropharyngeal swab samples from hospitalized patients (children < 12 years and adults) who had SARI within 7 days of the onset of symptoms, including fever (> 38.8 °C), dyspnea, and cough. Clinical and epidemiological information was obtained through standardized questionnaire. Enrolled patients were initially suspected to have influenza A(H1N1)pdm09 infections. WUPyV and KIPyV were detected by real-time PCR. Samples were also tested for influenza A and B viruses, human respiratory syncytial virus, rhinovirus, metapneumovirus, coronavirus, adenovirus, and parainfluenza viruses. WUPyV and KIPyV were detected in 6.77% (4.78% and 1.99%, respectively) of hospitalized patients with SARI. All samples from children showed coinfections (rhinovirus was the most commonly detected). Six adults had polyomavirus infection and four (1.6%) had monoinfection. Of them, 3 reported comorbidities including immunosuppression and 1 patient had worse outcome, requiring ICU admission. These preliminary data may suggest a possible role of polyomaviruses in SARI among immunocompromised adult patients.


title: Just transitions: Histories and futures in a post-COVID world

text: The energy landscape is changing dramatically. Communities are being impacted in different ways. Positive impacts include reductions in air pollution and new tax revenues from renewables. Negative impacts include lost jobs and foregone tax revenues after closure of large fossil fuels generation facilities and coal mines. The contours of this transition have been further altered by recent events such as the global oil market crash and the COVID-19 pandemic. While economic and social issues can be addressed through thoughtful policy design, the pace of change, and the extent to which communities have a say in what comes next, matter. Though the technical issues of transitions are well-researched, the socio-economic aspects of the energy transition remain both emergent and essential to an equitable transition to a low-carbon energy system. This article provides an overview of the history and current status of just transitions.


Pivoting Research to COVID‐19


title: A Modified Coupled Spectrophotometric Method to Detect 2-5 Oligoadenylate Synthetase Activity in Prostate Cell Lines

text: BACKGROUND: 2’-5’ oligoadenylate synthetases (OAS) are interferon inducible enzymes that polymerizes ATP to 2’-5’-linked oligomers of adenylate (2-5As). As part of the innate immune response, these enzymes are activated by viral double stranded RNA or mRNAs with significant double stranded structure. The 2-5As in turn activate RNaseL that degrade single stranded RNAs. Three distinct forms of OAS exist in human cells (OAS1, 2 and 3) with each form having multiple spliced variants. The OAS enzymes and their spliced variants have different enzyme activities. OAS enzymes also play a significant role in regulating multiple cellular processes such as proliferation and apoptosis. Moreover, Single nucleotide polymorphisms that alter OAS activity are also associated with viral infection, diabetes and cancer. Thus detection of OAS enzyme activity with a simple spectrophotometric method in cells will be important in clinical research. RESULTS: Here we propose a modified coupled spectrophotometric assay to detect 2-5 oligoadenylate synthetase (OAS) enzyme activity in prostate cell lines as a model system. The OAS enzyme from prostate cancer cell lysates was purified using Polyinosinic: polycytidylic acid (poly I:C) bound activated sepharose beads. The activated OAS enzyme eluted from Sepharose beads showed expression of p46 isoform of OAS1, generally considered the most abundant OAS isoform in elutes from DU14 cell line but not in other prostate cell line. In this assay the phosphates generated by the OAS enzymatic reaction is coupled with conversion of the substrate 2-amino-6-mercapto-7-methylpurine ribonucleoside (methylthioguanosine, a guanosine analogue; MESG) to a purine base product, 2-amino-6-mercapto-7-methylpurine and ribose1-phosphate via a catalyst purine nucleoside phosphorylase (phosphorylase) using a commercially available pyrophosphate kit. The absorbance of the purine base product is measured at 360 nm. The higher levels of phosphates detected in DU145 cell line indicates more activity of OAS in this prostate cancer cell line. CONCLUSION: The modified simple method detected OAS enzyme activity with sensitivity and specificity, which could help in detection of OAS enzymes avoiding the laborious and radioactive methods.


title: Molecular Mapping of the RNA Cap 2′-O-Methyltransferase Activation Interface between Severe Acute Respiratory Syndrome Coronavirus nsp10 and nsp16

text: Several protein-protein interactions within the SARS-CoV proteome have been identified, one of them being between non-structural proteins nsp10 and nsp16. In this work, we have mapped key residues on the nsp10 surface involved in this interaction. Alanine-scanning mutagenesis, bioinformatics, and molecular modeling were used to identify several “hot spots,” such as Val(42), Met(44), Ala(71), Lys(93), Gly(94), and Tyr(96), forming a continuous protein-protein surface of about 830 Å(2), bearing very conserved amino acids among coronaviruses. Because nsp16 carries RNA cap 2′-O-methyltransferase (2′O-MTase) activity only in the presence of its interacting partner nsp10 (Bouvet, M., Debarnot, C., Imbert, I., Selisko, B., Snijder, E. J., Canard, B., and Decroly, E. (2010) PLoS Pathog. 6, e1000863), functional consequences of mutations on this surface were evaluated biochemically. Most changes that disrupted the nsp10-nsp16 interaction without structural perturbations were shown to abrogate stimulation of nsp16 RNA cap 2′O-MTase activity. More strikingly, the Y96A mutation abrogates stimulation of nsp16 2′O-MTase activity, whereas Y96F overstimulates it. Thus, the nsp10-nsp16 interface may represent an attractive target for antivirals against human and animal pathogenic coronaviruses.


title: Genome structure and transcriptional regulation of human coronavirus NL63

text: BACKGROUND: Two human coronaviruses are known since the 1960s: HCoV-229E and HCoV-OC43. SARS-CoV was discovered in the early spring of 2003, followed by the identification of HCoV-NL63, the fourth member of the coronaviridae family that infects humans. In this study, we describe the genome structure and the transcription strategy of HCoV-NL63 by experimental analysis of the viral subgenomic mRNAs. RESULTS: The genome of HCoV-NL63 has the following gene order: 1a-1b-S-ORF3-E-M-N. The GC content of the HCoV-NL63 genome is extremely low (34%) compared to other coronaviruses, and we therefore performed additional analysis of the nucleotide composition. Overall, the RNA genome is very low in C and high in U, and this is also reflected in the codon usage. Inspection of the nucleotide composition along the genome indicates that the C-count increases significantly in the last one-third of the genome at the expense of U and G. We document the production of subgenomic (sg) mRNAs coding for the S, ORF3, E, M and N proteins. We did not detect any additional sg mRNA. Furthermore, we sequenced the 5' end of all sg mRNAs, confirming the presence of an identical leader sequence in each sg mRNA. Northern blot analysis indicated that the expression level among the sg mRNAs differs significantly, with the sg mRNA encoding nucleocapsid (N) being the most abundant. CONCLUSIONS: The presented data give insight into the viral evolution and mutational patterns in coronaviral genome. Furthermore our data show that HCoV-NL63 employs the discontinuous replication strategy with generation of subgenomic mRNAs during the (-) strand synthesis. Because HCoV-NL63 has a low pathogenicity and is able to grow easily in cell culture, this virus can be a powerful tool to study SARS coronavirus pathogenesis.


title: Otolaryngology Resident Practices and Perceptions in the Initial Phase of the U.S. COVID‐19 Pandemic

text: OBJECTIVE: The coronavirus 2019 (COVID‐19) pandemic has had widespread implications on clinical practice at U.S. hospitals. These changes are particularly relevant to otolaryngology–head and neck surgery (OHNS) residents because reports suggest an increased risk of contracting COVID‐19 for otolaryngologists. The objectives of this study were to evaluate OHNS residency program practice changes and characterize resident perceptions during the initial phase of the pandemic. STUDY DESIGN: A cross‐sectional survey of U.S. OHNS residents at 81 programs was conducted between March 23, 2020, and March 29, 2020. RESULTS: Eighty‐two residents from 51 institutions (63% of invited programs) responded. At the time of survey, 98% of programs had enacted policy changes to minimize COVID‐19 spread. These included filtered respirator use for aerosol‐generating procedures even in COVID‐19‐negative patients (85%), decreased resident staffing of surgeries (70%), and reduced frequency of tracheotomy care (61%). The majority of residents (66%) perceived that residents were at higher risk of contracting COVID‐19 compared to attendings. Residents were most concerned about protective equipment shortage (93%) and transmitting COVID‐19 to patients (90%). The majority of residents (73%) were satisfied with their department's COVID‐19 response. Resident satisfaction correlated with comfort level in discussing concerns with attendings (r = 0.72, P < .00001) and inversely correlated with perceptions of increased risk compared to attendings (r = −0.52, P < .00001). CONCLUSION: U.S. OHNS residency programs implemented policy changes quickly in response to the COVID‐19 pandemic. Sources of resident anxieties demonstrate the importance of open communication and an integrated team approach to facilitate optimal patient and provider care during this unprecedented crisis. LEVEL OF EVIDENCE: 4. Laryngoscope, 2020


title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

text: The association of Zika virus (ZIKV) infection and severe complications including neurological sequelae especially fetal microcephaly has aroused global attentions since its outbreak in 2015. Currently, there are no vaccines or therapeutic drugs clinically approved for treatments of ZIKV infection, however. And the drugs used for treating ZIKV in pregnant women require a higher safety profile. Here, we identified an anti-asthmatic drug, montelukast, which is of safety profile for pregnant women and exhibited antiviral efficacy against ZIKV infection in vitro and in vivo. And we showed that montelukast could disrupt the integrity of the virions to release the viral genomic RNA, hence irreversibly inhibiting viral infectivity. In consideration of the neuro-protective activity that montelukast possessed, which was previously reported, it is promising that montelukast could be used for patients with ZIKV infection, particularly for pregnant women.


title: G0/G1 arrest and apoptosis induced by SARS-CoV 3b protein in transfected cells

text: Severe Acute Respiratory Syndrome coronavirus (SARS-CoV), cause of the life-threatening atypical pneumonia, infects many organs, such as lung, liver and immune organ, and induces parenchyma cells apoptosis and necrosis. The genome of SARS-CoV, not closely related to any of the previously characterized coronavirus, encodes replicase and four major structural proteins and a number of non-structural proteins. Published studies suggest that some non-structural proteins may play important roles in the replication, virulence and pathogenesis of viruses. Among the potential SARS-CoV non-structural proteins, 3b protein (ORF4) is predicted encoding 154 amino acids, lacking significant similarities to any known proteins. Till now, there is no report about the function of 3b protein. In this study, 3b gene was linked with the EGFP tag at the C- terminus. Through cell cycle analysis, it was found that over-expression of 3b-EGFP protein in Vero, 293 and COS-7 cells could induce cell cycle arrest at G0/G1 phase, and that especially in COS-7 cells, expression of 3b-EGFP was able to induce the increase of sub-G1 phase from 24 h after transfection, which was most obvious at 48 h. The apoptosis induction of 3b fusion protein in COS-7 cells was further confirmed by double cell labeling with 7-AAD and Annexin V, the function of 3b protein inducing cell G0/G1 arrest and apoptosis may provide a new insight for further study on the mechanism of SARS pathogenesis.


title: Emerging viral diseases of fish and shrimp

text: The rise of aquaculture has been one of the most profound changes in global food production of the past 100 years. Driven by population growth, rising demand for seafood and a levelling of production from capture fisheries, the practice of farming aquatic animals has expanded rapidly to become a major global industry. Aquaculture is now integral to the economies of many countries. It has provided employment and been a major driver of socio-economic development in poor rural and coastal communities, particularly in Asia, and has relieved pressure on the sustainability of the natural harvest from our rivers, lakes and oceans. However, the rapid growth of aquaculture has also been the source of anthropogenic change on a massive scale. Aquatic animals have been displaced from their natural environment, cultured in high density, exposed to environmental stress, provided artificial or unnatural feeds, and a prolific global trade has developed in both live aquatic animals and their products. At the same time, over-exploitation of fisheries and anthropogenic stress on aquatic ecosystems has placed pressure on wild fish populations. Not surprisingly, the consequence has been the emergence and spread of an increasing array of new diseases. This review examines the rise and characteristics of aquaculture, the major viral pathogens of fish and shrimp and their impacts, and the particular characteristics of disease emergence in an aquatic, rather than terrestrial, context. It also considers the potential for future disease emergence in aquatic animals as aquaculture continues to expand and faces the challenges presented by climate change.


title: Impact of a multiplex PCR point-of-care test for influenza A/B and respiratory syncytial virus on an acute pediatric hospital ward

text: Abstract Patients with respiratory infections are often managed presumptively until confirmation of infection status. We assessed the impact of introducing the Enigma® MiniLab™ FluAB-RSV point-of-care test (POCT) on patients admitted with a suspected respiratory virus driven illness in an acute pediatric ward. This utilized a before and after design (respiratory viral seasons 2013/14 versus 2014/15). Following POCT implementation, oseltamivir prescribing increased in patients with influenza (OR = 12.7, P = 0.05, 95% CI [1.0, 153.8]). A reduction in the average reimbursement charges without a change in the length of stay was observed. Modeling suggested that laboratory test cost savings could be achieved if the POCT cost £30 and was used for screening, followed by the respiratory viral panel for RSV and influenza negative patients. A rapid POCT for influenza A/B and RSV infections in pediatric inpatients may improve oseltamivir prescribing, strengthen antimicrobial stewardship, reduce reimbursement charges and decrease laboratory costs.


title: Characterization by high-resolution crystal structure analysis of a triple-helix region of human collagen type III with potent cell adhesion activity

text: Abstract Collagen is one of the most abundant and important proteins in the human body. Human collagen type III (hCOL3A1) belongs to the fibril-forming collagens and is widely distributed in extensible connective tissue like skin, internal organs, or the vascular system. It plays key roles in wound healing, collagen fibrillogenesis, and normal cardiovascular development in human. The charged residues are considered to be an important characteristic of hCOL3A1, especially for collagen binding and recognition. Here we found that a triple helix fragment of hCOL3A1, Gly489-Gly510, contained multiple charged residues, as well as representative Glu-Lys-Gly and Glu-Arg-Gly charged triplets. We solved the crystal structure of this new fragment to a high-resolution of 1.50 Å and identified some important conformations of this new triple-helix region, including strong hydrogen bonds in interchain and interhelical interactions in addition to obvious flexible bending for the triple helix. We also found that the synthetic collagen peptides around this region exhibited potent activities through integrin-mediated peptide-membrane interaction. We then developed a method to produce a recombinant protein consisting of 16 tandem repeats of the triple-helix fragment of hCOL3A1 with strong activity without cytotoxicity. These results provide a strong base for further functional studies of human collagen type III and the method developed in this study can be applied to produce hCOL3A1-derived proteins or other tandem-repeat proteins with membrane adhesion activity.


title: Planning horizon affects prophylactic decision-making and epidemic dynamics

text: The spread of infectious diseases can be impacted by human behavior, and behavioral decisions often depend implicitly on a planning horizon—the time in the future over which options are weighed. We investigate the effects of planning horizons on epidemic dynamics. We developed an epidemiological agent-based model (along with an ODE analog) to explore the decision-making of self-interested individuals on adopting prophylactic behavior. The decision-making process incorporates prophylaxis efficacy and disease prevalence with the individuals’ payoffs and planning horizon. Our results show that for short and long planning horizons individuals do not consider engaging in prophylactic behavior. In contrast, individuals adopt prophylactic behavior when considering intermediate planning horizons. Such adoption, however, is not always monotonically associated with the prevalence of the disease, depending on the perceived protection efficacy and the disease parameters. Adoption of prophylactic behavior reduces the epidemic peak size while prolonging the epidemic and potentially generates secondary waves of infection. These effects can be made stronger by increasing the behavioral decision frequency or distorting an individual’s perceived risk of infection.


title: Viral Loads and Duration of Viral Shedding in Adult Patients Hospitalized with Influenza

text: BackgroundThe goal of this study was to characterize viral loads and factors affecting viral clearance in persons with severe influenza MethodsThis was a 1-year prospective, observational study involving consecutive adults hospitalized with influenza. Nasal and throat swabs were collected at presentation, then daily until 1 week after symptom onset. Real-time reverse-transcriptase polymerase chain reaction to determine viral RNA concentration and virus isolation were performed. Viral RNA concentration was analyzed using multiple linear or logistic regressions or mixed-effect models ResultsOne hundred forty-seven inpatients with influenza A (H3N2) infection were studied (mean age ± standard deviation, 72±16 years). Viral RNA concentration at presentation positively correlated with symptom scores and was significantly higher than that among time-matched outpatients (control subjects). Patients with major comorbidities had high viral RNA concentration even when presenting >2 days after symptom onset (mean ± standard deviation, 5.06±1.85 vs 3.62±2.13 log(10) copies/mL; P=.005; β, +0.86 [95% confidence interval, +0.03 to +1.68]). Viral RNA concentration demonstrated a nonlinear decrease with time; 26% of oseltamivir-treated and 57% of untreated patients had RNA detected at 1 week after symptom onset. Oseltamivir started on or before symptom day 4 was independently associated with an accelerated decrease in viral RNA concentration (mean β [standard error], −1.19 [0.43] and −0.68 [0.33] log(10) copies/mL for patients treated on day 1 and days 2–3, respectively; P<.05) and viral RNA clearance at 1 week (odds ratio, 0.10 [95% confidence interval, 0.03–0.35] and 0.30 [0.10–0.90] for patients treated on day 1–2 and day 3–4, respectively). Conversely, major comorbidities and systemic corticosteroid use for asthma or chronic obstructive pulmonary disease exacerbations were associated with slower viral clearance. Viral RNA clearance was associated with a shorter hospital stay (7.0 vs 13.5 days; P=.001) ConclusionPatients hospitalized with severe influenza have more active and prolonged viral replication. Weakened host defenses slow viral clearance, whereas antivirals started within the first 4 days of illness enhance viral clearance


title: Advanced renal cell carcinoma and COVID-19 — a personal perspective

text: The management pathways of advanced renal cell carcinoma (RCC) have considerably evolved in the past 5 years, presenting a particular challenge during the coronavirus disease 2019 (COVID-19) pandemic. In this Comment, we propose a management algorithm on the basis of the current evidence to safely manage patients with advanced RCC during this pandemic.


title: Virtual screening for the discovery of bioactive natural products

text: In this survey the impact of the virtual screening concept is discussed in the field of drug discovery from nature. Confronted by a steadily increasing number of secondary metabolites and a growing number of molecular targets relevant in the therapy of human disorders, the huge amount of information needs to be handled. Virtual screening filtering experiments already showed great promise for dealing with large libraries of potential bioactive molecules. It can be utilized for browsing databases for molecules fitting either an established pharmacophore model or a three dimensional (3D) structure of a macromolecular target. However, for the discovery of natural lead candidates the application of this in silico tool has so far almost been neglected. There are several reasons for that. One concerns the scarce availability of natural product (NP) 3D databases in contrast to synthetic libraries; another reason is the problematic compatibility of NPs with modern robotized high throughput screening (HTS) technologies. Further arguments deal with the incalculable availability of pure natural compounds and their often too complex chemistry. Thus research in this field is time-consuming, highly complex, expensive and ineffective. Nevertheless, naturally derived compounds are among the most favorable source of drug candidates. A more rational and economic search for new lead structures from nature must therefore be a priority in order to overcome these problems. Here we demonstrate some basic principles, requirements and limitations of virtual screening strategies and support their applicability in NP research with already performed studies. A sensible exploitation of the molecular diversity of secondary metabolites however asks for virtual screening concepts that are interfaced with well-established strategies from classical pharmacognosy that are used in an effort to maximize their efficacy in drug discovery. Such integrated virtual screening workflows are outlined here and shall help to motivate NP researchers to dare a step towards this powerful in silico tool.


title: Studying immunity to zoonotic diseases in the natural host — keeping it real

text: Zoonotic viruses that emerge from wildlife and domesticated animals pose a serious threat to human and animal health. In many instances, mouse models have improved our understanding of the human immune response to infection; however, when dealing with emerging zoonotic diseases, they may be of limited use. This is particularly the case when the model fails to reproduce the disease status that is seen in the natural reservoir, transmission species or human host. In this Review, we discuss how researchers are placing more emphasis on the study of the immune response to zoonotic infections in the natural reservoir hosts and spillover species. Such studies will not only lead to a greater understanding of how these infections induce variable disease and immune responses in distinct species but also offer important insights into the evolution of mammalian immune systems. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nri3551) contains supplementary material, which is available to authorized users.


title: Proventricular dilatation disease: an emerging exotic disease of parrots in Australia

text: Proventricular dilatation disease is a viral disease seen as a segmental neuropathy in parrots. It has always been believed to be a disease exotic to Australia, with the only reported case being a legally imported Green Wing Macaw (Ara chloroptera) in 1993. This paper reports a cluster of cases seen in south‐east Queensland in 2005 to 2006. Clinical signs, autopsy findings and histopathological findings are described. No pattern or common source for these cases could be identified. The implications for Australian aviculture and avifauna are discussed.


title: Response to the 2009 pandemic: Effect on influenza control in wealthy and poor countries

text: The declaration by the World Health Organization (WHO) that appearance of a swine-origin novel influenza virus in 2009 represented a pandemic was based on previously adopted guidelines and the new International Health Regulations. Severity of the pandemic was not part of the definition used, but it was stated to be less than severe at the time of declaration. It was necessary, when there was still uncertainty about the overall impact of the pandemic, for vaccine production to begin to have timely availability. Countries arranged to have vaccine for their populations, and WHO attempted to secure supplies for under-resourced countries. The world had been concerned that the next pandemic might be a severe one, based on the specter of avian influenza with a case fatality of up to 80% in humans. After it was clear that the 2009 pandemic was not severe, there were accusations, especially in Europe, that countries had secured vaccine supplies mainly to benefit the manufacturers. Such charges, even when refuted, may undermine public confidence in the process which assures vaccine supply and availability of vaccine for seasonal use. Production of pandemic vaccine is conditioned on the supply of seasonal influenza vaccine; it is unrealistic to expect vaccine to be available for pandemic use when none is used for seasonal influenza. This particularly applies to poorer counties. They have traditionally not recognized that influenza is a problem, although this attitude is changing. As we go forward, we need to keep in mind the global nature of the threat of influenza. Had the 2009 pandemic been more severe, demand would have been greater and poorer counties would have had little vaccine to meet their needs. Only by taking a broad view of influenza on an annual basis can vaccine supplies be ensured for all countries of the world.


title: Adoptive Transfer of Infectious Bronchitis Virus Primed αβ T Cells Bearing CD8 Antigen Protects Chicks from Acute Infection

text: Abstract Infectious bronchitis virus (IBV) infection and associated illness may be dramatically modified by passive transfer of immune T lymphocytes. Lymphocytes collected 10 days postinfection were transferred to naive chicks before challenge with virus. As determined by respiratory illness and viral load, transfer of syngeneic immune T lymphocytes protected chicks from challenge infection, whereas no protection was observed in the chicks receiving the MHC compatible lymphocytes from uninfected chicks. Protection following administration of T lymphocytes could be observed in chicks with three distinct MHC haplotypes: B8 /B8, B12 /B12, and B19 /B19 . Nearly complete elimination of viral infection and illness was observed in chicks receiving cells enriched in αβ lymphocytes. In contrast, removal of γδ T lymphocytes had only a small effect on their potential to protect chicks. The adoptive transfer of enriched CD8+ or CD4+ T lymphocytes indicated that protection was also a function primarily of CD8-bearing cells. These results indicated that αβ T lymphocytes bearing CD8+ antigens are critical in protecting chicks from IBV infection.


title: Letter: does vitamin D have a potential role against COVID‐19? Authors' reply

text: We thank Kumar et al for their comments on our review article and the letter connected with that by Panarese and Shahini.(1,2) We agree that there is a complicated effect of vitamin D in preventing the severity of COVID‐19, while this mechanism is not exactly the same as that of influenza.


title: Experimentally confirmed toltrazuril resistance in a field isolate of Cystoisospora suis

text: BACKGROUND: Constant treatment regimens with toltrazuril against Cystoisospora suis infection in piglets are being applied in the intensive production systems for the last two decades, but the possibility of resistance development has not been addressed so far despite limited availability of treatment alternatives. Recently, a pig producer in The Netherlands who routinely used toltrazuril complained about diarrhea in suckling piglets in the absence of bacterial and viral pathogens, and oocysts of C. suis could be isolated from feces of affected litters. METHODS: Piglets from two litters were infected with a field isolate of C. suis, Holland-I, and treated with 0 (Holl-Ctrl), 20 (Holl-20) or 30 (Holl-30) mg/kg of body weight (BW) of toltrazuril (Baycox®). The efficacy of toltrazuril was measured by assessment of oocyst excretion, fecal consistency and BW gain. A separate litter was infected with a toltrazuril-susceptible strain of C. suis, Wien-I, and treated with 0 (Wien-Ctrl) or 20 (Wien-20) mg/kg BW of toltrazuril for comparison. RESULTS: Treatment with the recommended (20 mg/kg) dose of toltrazuril completely suppressed oocyst shedding and diarrhea in group Wien-20. The prevalence of oocyst excretion was 100% in the groups infected with Holland-I and 80% in the group Wien-Ctrl. Most days with diarrhea were observed in group Holl-20 with an average of 6.40%, followed by 5.71% in Wien-Ctrl, while in Holl-Ctrl and Holl-30 diarrhea was only seen in 1.79% of the samples (n = 14/piglet). Oocyst excretion, fecal consistency and BW gain did not differ significantly among groups infected with Holland-I, indicating loss of efficacy to toltrazuril. CONCLUSION: Experimental infections and treatment confirmed toltrazuril resistance against the field isolate even at increased dosage. Such isolates are a potential threat to pig production as no other effective and economically sustainable alternative treatment is currently available. In the absence of a standardized protocol for resistance testing in C. suis, regular parasitological examination and, if possible, experimental confirmation should be considered to evaluate the extent and consequences of toltrazuril resistance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-017-2257-7) contains supplementary material, which is available to authorized users.


title: Evaluation of visual triage for screening of Middle East respiratory syndrome coronavirus patients

text: The emergence of Middle East respiratory syndrome coronavirus (MERS-CoV) in September 2012 in Saudi Arabia had attracted the attention of the global health community. In 2017 the Saudi Ministry of Health released a visual triage system with scoring to alert healthcare workers in emergency departments (EDs) and haemodialysis units for the possibility of occurrence of MERS-CoV infection. We performed a retrospective analysis of this visual score to determine its sensitivity and specificity. The study included all cases from 2014 to 2017 in a MERS-CoV referral centre in Riyadh, Saudi Arabia. During the study period there were a total of 2435 suspected MERS cases. Of these, 1823 (75%) tested negative and the remaining 25% tested positive for MERS-CoV by PCR assay. The application of the visual triage score found a similar percentage of MERS-CoV and non–MERS-CoV patients, with each score from 0 to 11. The percentage of patients with a cutoff score of ≥4 was 75% in patients with MERS-CoV infection and 85% in patients without MERS-CoV infection (p 0.0001). The sensitivity and specificity of this cutoff score for MERS-CoV infection were 74.1% and 18.6%, respectively. The sensitivity and specificity of the scoring system were low, and further refinement of the score is needed for better prediction of MERS-CoV infection.


title: Specific EEG Encephalopathy Pattern in SARS-CoV-2 Patients

text: We used quantified electroencephalography (qEEG) to define the features of encephalopathy in patients released from the intensive care unit after severe illness from COVID-19. Artifact-free 120–300 s epoch lengths were visually identified and divided into 1 s windows with 10% overlap. Differential channels were grouped by frontal, parieto-occipital, and temporal lobes. For every channel and window, the power spectrum was calculated and used to compute the area for delta (0–4 Hz), theta (4–8 Hz), alpha (8–13 Hz), and beta (13–30 Hz) bands. Furthermore, Shannon’s spectral entropy (SSE) and synchronization by Pearson’s correlation coefficient (ρ) were computed; cases of patients diagnosed with either infectious toxic encephalopathy (ENC) or post-cardiorespiratory arrest (CRA) encephalopathy were used for comparison. Visual inspection of EEGs of COVID patients showed a near-physiological pattern with scarce anomalies. The distribution of EEG bands was different for the three groups, with COVID midway between distributions of ENC and CRA; specifically, temporal lobes showed different distribution for EEG bands in COVID patients. Besides, SSE was higher and hemispheric connectivity lower for COVID. We objectively identified some numerical EEG features in severely ill COVID patients that can allow positive diagnosis of this encephalopathy.


title: Higher levels of stress and different coping strategies are associated with greater morning and evening fatigue severity in oncology patients receiving chemotherapy

text: PURPOSE: A cancer diagnosis and associated treatments are stressful experiences for most patients. Patients’ perceptions of stress and their use of coping strategies may influence fatigue severity. This study extends our previous work describing distinct profiles of morning (i.e., Very Low, Low, High, and Very High) and evening (i.e., Low, Moderate, High, and Very High) fatigue in oncology patients by evaluating for differences in stress and coping strategies among these fatigue classes. METHODS: This longitudinal study evaluated for changes in morning and evening fatigue in oncology patients (n = 1332) over two cycles of chemotherapy (CTX). Patients completed measures of cumulative exposure to stressful life events (SLEs) (i.e., the Life Stressor Checklist-Revised), general stress (i.e., Perceived Stress Scale [PSS]), cancer-specific stress (i.e., Impact of Event Scale-Revised [IES-R]), and coping strategies (i.e., Brief Cope). Differences among the latent classes were evaluated using analyses of variance, Kruskal-Wallis, or chi-square tests. RESULTS: Patients in both the Very High morning and evening fatigue classes reported higher numbers of and a higher impact from previous SLEs and higher PSS scores than the other fatigue classes. The IES-R scores for the Very High morning fatigue class met the criterion for subsyndromal PTSD. Patients in the Very High evening fatigue class used a higher number of engagement coping strategies compared with the Very High morning fatigue class. CONCLUSIONS: Our findings suggest that interventions to reduce stress and enhance coping warrant investigation to decrease fatigue in patients undergoing CTX. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00520-020-05303-5) contains supplementary material, which is available to authorized users.


title: Engineered Protein Model of the ATP synthase H(+)- Channel Shows No Salt Bridge at the Rotor-Stator Interface

text: ATP synthase is powered by the flow of protons through the molecular turbine composed of two α-helical integral membrane proteins, subunit a, which makes a stator, and a cylindrical rotor assembly made of multiple copies of subunit c. Transient protonation of a universally conserved carboxylate on subunit c (D61 in E. coli) gated by the electrostatic interaction with arginine on subunit a (R210 in E. coli) is believed to be a crucial step in proton transfer across the membrane. We used a fusion protein consisting of subunit a and the adjacent helices of subunit c to test by NMR spectroscopy if cD61 and aR210 are involved in an electrostatic interaction with each other, and found no evidence of such interaction. We have also determined that R140 does not form a salt bridge with either D44 or D124 as was suggested previously by mutation analysis. Our results demonstrate the potential of using arginines as NMR reporter groups for structural and functional studies of challenging membrane proteins.


title: In Vitro synthesis of two polypeptides from a nonstructural gene of coronavirus mouse hepatitis virus strain A59

text: Abstract The complete nucleotide sequence of nonstructural gene 5 of coronavirus mouse hepatitis virus (MHV) strain A59 has been determined. This sequence contains two potential open reading frames which overlap by five nucleotides. The putative protein products predicted from the sequence are a basic 13,000-Da polypeptide and a 9600-Da polypeptide containing an unusually long hydrophobic amino terminus. RNAs transcribed in vitro from DNAs containing each of the open reading frames in pGEM vectors direct the synthesis in vitro of polypeptides of the sizes predicted by the sequence. An RNA transcript containing both of the open reading frames directs the synthesis primarily of the polypeptide corresponding to the downstream open reading frame. These data suggest that MHV-A59 messenger RNA 5 potentially encodes two proteins and may be preferentially translated from an internal AUG initiation codon.


title: Structure of the SARS coronavirus main proteinase as an active C(2) crystallographic dimer

text: The 34 kDa main proteinase (M(pro)) from the severe acute respiratory syndrome coronavirus (SARS-CoV) plays an important role in the virus life cycle through the specific processing of viral polyproteins. As such, SARS-CoV M(pro) is a key target for the identification of specific inhibitors directed against the SARS virus. With a view to facilitating the development of such compounds, crystals were obtained of the enzyme at pH 6.5 in the orthorhombic space group P2(1)2(1)2 that diffract to a resolution of 1.9 Å. These crystals contain one monomer per asymmetric unit and the biologically active dimer is generated via the crystallographic twofold axis. The conformation of the catalytic site indicates that the enzyme is active in the crystalline form and thus suitable for structure-based inhibition studies.


title: Hypothesis links emergence of chloroquine-resistant malaria and other intracellular pathogens and suggests a new strategy for treatment of diseases caused by intracellular parasites

text: Abstract Chloroquine and related anti-malarial drugs appear to promote apoptosis in T-cells by suppressing NF-kappa-B, which enhances the expression of anti-apoptotic proteins (e.g., Bcl-2). Thus, chloroquine has found applications in autoimmune diseases where it apparently facilitates apoptosis of abnormally persistent T-cell clones. The mode of action of chloroquine in prevention of malaria is not known, but it may be to minimize replication of the parasite in the liver cells, which occurs before invasion of the erythrocytes, by facilitating premature apoptosis of the infected host cells. After introduction of chloroquine in the 1950s world-wide for prophylactic use, chloroquine-resistant malaria emerged. Here it is hypothesized that concurrent with emergence of chloroquine-resistant malaria (presumably with enhanced anti-apoptotic capabilities), other intracellular parasites have evolved to enhance their ability to prevent apoptosis in host cells. Two examples of viral diseases that have emerged from areas of high incidence of chloroquine-resistant malaria are AIDS from HIV and SARS from coronavirus. The hypothesis holds that prophylactic exposure to pro-apoptotic chloroquine drugs caused natural selection for strains of viruses and other parasites that have enhanced anti-apoptotic abilities. When transmitted to host organisms that are not under the influence of the pro-apoptotic drug, the new “anti-apoptotic” strains may cause unexpected diseases. In the case of SARS, the coronavirus appears to have accessed a new niche where it proves to be lethal to its host. In the case of AIDS, the HIV (which has had a long-term symbiotic relationship with primates) has run amuck because the infected cells are now substantially more tolerant to the toxins (i.e., resistant to apoptosis) that they secrete than the uninfected bystander cells, which are not unusually resistant to apoptosis. A corollary to the hypothesis is that if the level of resistance to apoptosis in the infected cells were no higher than the level of resistance in the bystander cells, then the infected cells would preferentially kill themselves through apoptosis. It appears that in the case of HIV, the increased resistance to apoptosis is provided by expression of Bcl-2 and suppression of p53. Hence, drugs that suppresses Bcl-2 or restore p53 function might be effective in restoring the parity of resistance to apoptosis between infected and uninfected cells. Currently, an antisense drug targeting Bcl-2 (G3139/GenasenseTM, Genta, Inc.) is in late-stage cancer trials and may be on the market for those indications in months. It would be interesting to try these drugs against various intracellular parasites including HIV. This approach to prevent or eliminate active infections might be particularly attractive against a range of parasites (virus, bacteria, protozoa, fungus) when safe and effective vaccines are not available.


title: Antigen detection in human respiratory Coronavirus infections by monoclonal time-resolved fluoroimmunoassay

text: Abstract Background: The diagnosis of respiratory infections by detecting viral antigens has received considerable attention using immunofluorescent assays (IFA) and enzyme immunoassays (EIA). Time-resolved fluoroimmunoassay (TR-FIA) has been developed for several viruses. Objectives: To prepare monoclonal antibodies to coronavirus strains, to incorporate them into a TR-FIA, and test the assay on clinical specimens. Study design: Monoclonal antibodies were prepared to the N nucleoprotein of the two human respiratory coronaviruses, HCV strains 229E and OC43. Monoclonals to both viruses were completely type-specific; they did not cross-react between themselves or with multiple strains of other respiratory viruses. These antibodies were configured into optimized EIA and TR-FIA tests. The all-monoclonal tests were then compared to polyclonal EIA tests in terms of their ability to detect virus in clinical specimens. Results: The all-monoclonal TR-FIA was uniformly the most sensitive, detecting virus in all 13 229E-positive specimens compared to 69% for the monoclonal EIA and 54% for the polyclonal EIA test. Similar results were obtained for 10 OC43-positive specimens: 100% in TR-FIA, 90% in monoclonal EIA, and 80% in polyclonal EIA. For 229E in TR-FIA, mean positive/negative (P/N) ratios were 143 for 229E-positive human embryonic lung fibroblast (HLF) cell culture fluids and 10 for positive nasopharyngeal aspirate specimens; for OC43 in TR-FIA, mean P/N values were 964 for OC43-positive rhabdomyosarcoma (RD) cell culture fluids and 174 for positive NPA specimens. The sensitivities of the TR-FIA were determined with purified virions to be 0.308 ng virus per well for HCV-229E and 0.098 ng virus per well for HCV-OC43. Conclusions: This rapid and sensitive test appears to be much more sensitive than traditional antigen detection assays but will require more extensive field testing on clinical specimens.


title: Original Chemical Series of Pyrimidine Biosynthesis Inhibitors That Boost the Antiviral Interferon Response

text: De novo pyrimidine biosynthesis is a key metabolic pathway involved in multiple biosynthetic processes. Here, we identified an original series of 3-(1H-indol-3-yl)-2,3-dihydro-4H-furo[3,2-c]chromen-4-one derivatives as a new class of pyrimidine biosynthesis inhibitors formed by two edge-fused polycyclic moieties. We show that identified compounds exhibit broad-spectrum antiviral activity and immunostimulatory properties, in line with recent reports linking de novo pyrimidine biosynthesis with innate defense mechanisms against viruses. Most importantly, we establish that pyrimidine deprivation can amplify the production of both type I and type III interferons by cells stimulated with retinoic acid-inducible gene 1 (RIG-I) ligands. Altogether, our results further expand the current panel of pyrimidine biosynthesis inhibitors and illustrate how the production of antiviral interferons is tightly coupled to this metabolic pathway. Functional and structural similarities between this new chemical series and dicoumarol, which was reported before to inhibit pyrimidine biosynthesis at the dihydroorotate dehydrogenase (DHODH) step, are discussed.


title: A Tale of Two Viruses: Does Heterologous Flavivirus Immunity Enhance Zika Disease?

text: The rise of Zika virus (ZIKV) and its unusual clinical manifestations provided ground for speculative debate. The clinical severity of secondary dengue virus (DENV) infections is associated with antibody-dependent enhancement (ADE), and it was recently suggested that previous exposure to DENV may worsen ZIKV clinical outcomes. In this Opinion article we analyze the relationship among different flaviviruses and ADE. We discuss new evidence obtained in non-human primates and human cohorts demonstrating that there is no correlation to ADE when ZIKV infection occurs in the presence of pre-existing DENV immunity. We propose a redefinition of ADE in the context of complex immunological flavivirus interactions to provide a more objective perspective when translating in vitro or in vivo observations into the clinical setting.


title: Effect of single- versus double-gloving on virus transfer to health care workers’ skin and clothing during removal of personal protective equipment

text: BACKGROUND: The removal of personal protective equipment (PPE) after patient care may result in transfer of virus to hands and clothing of health care workers (HCWs). The risk of transfer can be modeled using harmless viruses to obtain quantitative data. To determine whether double-gloving reduces virus transfer to HCWs’ hands and clothing during removal of contaminated PPE, we conducted a human challenge study using bacteriophages to compare the frequency and quantity of virus transfer to hands and clothes during PPE removal with single-gloving and double-gloving technique. METHODS: Each experiment had a double-gloving phase and a single-gloving phase. Participants donned PPE (ie, contact isolation gown, N95 respirator, eye protection, latex gloves). The gown, respirator, eye protection, and dominant glove were contaminated with bacteriophage. Participants then removed the PPE, and their hands, face, and scrubs were sampled for virus. RESULTS: Transfer of virus to hands during PPE removal was significantly more frequent with single-gloving than with double-gloving. Transfer to scrubs was similar during single-gloving and double-gloving. The amount of virus transfer to hands ranged from 0.15 to 2.5 log(10) most probable number. Significantly more virus was transferred to participants’ hands after single-gloving than after double-gloving. CONCLUSIONS: Our comparison of double-gloving and single-gloving using a simulation system with MS2 and a most-probable number method suggests that double gloving can reduce the risk of viral contamination of HCWs’ hands during PPE removal. If incorporated into practice when full PPE is worn, this practice may reduce the risk of viral contamination of HCWs’ hands during PPE removal. The use of double gloves should be explored in larger controlled studies.