title: Comparison of molecular detection methods for pertussis in children during a state-wide outbreak

text: A state-wide pertussis outbreak occurred in Washington during the winter–spring months of 2012, concurrent with respiratory viral season. We compared performance characteristics of a laboratory-developed pertussis PCR (LD-PCR for Bordetella pertussis, Bordetella parapertussis, and Bordetella holmesii) and rapid multiplex PCR (RM-PCR) for respiratory viruses (FilmArray™, BioFire, B. pertussis data unblinded following FDA approval post outbreak). We analyzed three cohorts of patients using physician testing orders as a proxy for clinical suspicion for pertussis or respiratory viruses: Cohort 1, tested by LD-PCR for pertussis pathogens only by nasopharyngeal swab; Cohort 2, by RM-PCR for respiratory viruses only by mid-nasal turbinate swab; and Cohort 3, by both methods. B. pertussis was detected in a total of 25 of the 490 patients in Cohort 3 in which LD-PCR detected 20/25 (80 %) cases and the RM-PCR detected 24/25 (96 %; p = 0.2). Pertussis pathogens were detected in 21/584 (3.6 %) of samples from Cohort 1 where clinicians had a relatively strong suspicion for pertussis. In contrast, B. pertussis was detected in only 4/3071 (0.1 %) specimens from Cohort 2 where suspicion for pertussis was lower (p < 0.001 for comparison with Cohort 1). In summary, the two laboratory methods were comparable for the detection of B. pertussis.


title: La Kinésithérapie face au COVID-19: un rôle de Santé publique majeur à court, moyen et long termes dans le processus rééducatif-réadaptatif des patients

text: Résumé Le coronavirus SRAS-CoV-2 est une zoonose virale à transmission interhumaine responsable d’une crise sanitaire mondiale. Les patients hospitalisés pour une pneumonie à SRAS-CoV-2 risquent de développer : une neuromyopathie, une sarcopénie, un déconditionnement à l’effort, un déséquilibre des comorbidités, une perte d’autonomie dans les activités de la vie quotidienne, une diminution de leur qualité de vie. En France, le masseur-kinésithérapeute joue un rôle majeur de Santé publique dans le traitement de ces patients en milieu hospitalier et à l’interface du système ville-hôpital-centre de rééducation fonctionnelle-domicile-établissement d'hébergement pour personnes âgées dépendantes. L’objectif de cet article est de faire une description scientifique pragmatique de l’attitude diagnostique et thérapeutique adoptée en pratique quotidienne par le masseur-kinésithérapeute dans le parcours de soin du patient affecté. Les conséquences bio-psycho-sociales de cette pandémie seront probablement importantes. La rééducation est déterminante pour le pronostic fonctionnel des patients atteints. À long terme, son objectif majeur sera la restauration multifonctionnelle. Abstract The SARS-CoV-2 coronavirus is a human-to-human viral zoonosis responsible for a global health crisis. Patients hospitalized with SARS-CoV-2 pneumonia are at risk of developing: neuromyopathy, sarcopenia, stress deconditioning, comorbidities imbalance, loss of autonomy in activities of daily living, reduced quality of life. In France, the physiotherapist plays a major public health role in the treatment of these patients in hospitals, rehabilitation centres and nursing homes. The main objective of this article is to make a pragmatic scientific description of the diagnostic and therapeutic attitude adopted in daily practice by the physiotherapist in the care of affected patient. Bio-psycho-social consequences of this pandemic are likely to be significant. Rehabilitation is decisive for the functional prognosis of affected patients. In the long term, its major objective will be multifunctional restoration.


title: Delayed diagnosis of paediatric appendicitis during the COVID‐19 pandemic

text: AIM: To present seven paediatric patients with appendicitis, all with late diagnosis resulting from different aspects of the fear from the current global COVID‐19 pandemic. METHODS: Cases were collected from three paediatric surgical wards. Comparison between complicated appendicitis rates in the COVID‐19 era and similar period in previous year was performed. RESULTS: All seven children presented with complicated appendicitis. Main reasons for the delayed diagnosis during the COVID‐19 era were parental concern, telemedicine use and insufficient evaluation. Higher complication rates were found during the COVID‐19 era compared to similar period in previous year (22% vs 11%, P‐value .06). CONCLUSION: The fear from COVID‐19 pandemic may result in delayed diagnosis and higher complication rates in common paediatric medical conditions. We believe caregivers and healthcare providers should not withhold necessary medical care since delay in diagnosis and treatment in these routinely seen medical emergencies may become as big of a threat as COVID‐19 itself.


title: COVID 19 outbreak: organisation of a geriatric assessment and coordination unit. A French example

text: Older people are particularly affected by the COVID19 outbreak because of their vulnerability as well as the complexity of health organisations, particularly in the often-compartmentalised interactions between community, hospital and nursing home actors. In this endemic situation, with massive flows of patients requiring holistic management including specific and intensive care, the appropriate assessment of each patient’s level of care and the organisation of specific networks is essential. To use the basis of territorial organisation of health by facilitating the link between hospital settings and geriatric sectors at the regional level. To connect private, medico-social and hospital actors through a dedicated centralised unit for evaluation, geriatric coordination of care and decision support. A geriatrician coordinates this multidisciplinary unit. It includes an emergency room doctor, a supervisor from the medical regulation centre (Centre 15), an infectious disease physician, a medical hygienist and a palliative care specialist. To organise an ad hoc follow-up channel, including the necessary resources for the different levels of care required, according to the resources of the territorial network, and the creation of a specific COVID geriatric palliative care service. This organisation meets the urgent health needs of all stakeholders, facilitating its deployment and allows the sustainable implementation of a coordinated geriatric management dynamic between the stakeholders on the territory.


title: Otolaryngology-related Google Search trends during the COVID-19 pandemic

text: OBJECTIVE: To assess trends of Google Search queries for symptoms and complaints encountered commonly in otolaryngology practices during the coronavirus disease 2019 (COVID-19) pandemic when in-person care has been limited. MATERIALS AND METHODS: In this cross-sectional study, data on Google Search queries in the United States for 30 otolaryngology-related terms were obtained from Google Trends. The means of relative search volume from the COVID-19 period (March 29, 2020 through May 16, 2020) were compared to similar periods from 2016 to 2019 using a t-test of two independent samples. RESULTS: In total, 16.6% of search terms had significant increases in relative search volume during the COVID-19 period, with the largest percentage increase for “can't smell” (124.4%, p = .006), followed by “allergies” (30.3%, p = .03), “voice pain” (26.1%, p = .008), and “ears ringing” (19.0%, p < .001). Of all search terms, 26.7% had significant decreases in relative search volume, including the largest percentage decrease for “laryngitis” (59.8%, p < .001), followed by “thyroid nodule” (54.4%, p < .001), “thyroid cancer” (45.6%, p < .001), and “ENT” (34.9%, p < .001). CONCLUSION: This study demonstrates that Google search activity for many otolaryngology-related terms during the COVID-19 pandemic has increased or decreased significantly as compared to previous years. With reduced access to in-office otolaryngology care in the United States during the COVID-19 pandemic, these are important considerations for otolaryngology practices to meet the needs of patients who lack access to care.


title: Genetic diversity of avian infectious bronchitis virus California variants isolated between 1988 and 2001 based on the S1 subunit of the spike glycoprotein

text: Twenty-nine isolates of avian infectious bronchitis virus (IBV) recovered from commercial chicken flocks in California between 1988 and 2001 and identified as California variants by serotype and direct automated cycle sequencing of the IBV spike glycoprotein S1 subunit, were further characterized phylogenetically and by nucleotide sequence comparison. California variants were grouped according to production type of chicken, by comparison with public access sequence databases (NCBI GenBank and EMBL), or based on phylogenetic analysis. Fisher’s Exact test was used to compare mutations per year, purifying and positive selection, predictive antigenicity, and a ≥ 6 bp deletion between California variant groups. A high number of mutations at the nucleotide level (p = 0.013) and a ≥ 6 bp deletion in the nucleotide sequence (p = 0.006) was significantly associated with broiler-type chickens. However, 88% of significant comparisons at the amino acid level such as purifying and positive selection were seen in layer-type chickens. A pronounced predictive antigenicity in the HVR2 region was also associated with layer-type chickens (p = 0.001). The study indicates that IBV in California is in a phase of slow evolution with different evolutionary patterns being associated with the production type of chicken.


title: 2015 RAD-AID Conference on International Radiology for Developing Countries: The Evolving Global Radiology Landscape

text: Abstract Radiology in low- and middle-income (developing) countries continues to make progress. Research and international outreach projects presented at the 2015 annual RAD-AID conference emphasize important global themes, including (1) recent slowing of emerging market growth that threatens to constrain the advance of radiology, (2) increasing global noncommunicable diseases (such as cancer and cardiovascular disease) needing radiology for detection and management, (3) strategic prioritization for pediatric radiology in global public health initiatives, (4) continuous expansion of global health curricula at radiology residencies and the RAD-AID Chapter Network’s participating institutions, and (5) technologic innovation for recently accelerated implementation of PACS in low-resource countries.


Corrigendum to “Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and COVID-19 infection during pregnancy” [Trav Med Infect Dis. (2020) 101641]


title: Covid-19: A Dynamic Analysis of Fatality Risk in Italy

text: Italy was the second country in the world to face a wide epidemic of Covid-19 after China. The ratio of the number of fatalities to the number of cases (case fatality ratio, CFR) recorded in Italy was surprisingly high and increased in the month of March. The older mean age of population, the changes in testing policy, and the methodological computation of CFR were previously reported as possible explanations for the incremental trend of CFR, a parameter theoretically expected to be constant. In this brief report, the official data provided by the Italian Ministry of Health were analyzed using fitting models and the linear fit method approach. This last methodology allowed us to reach two findings. The trend of the number of deaths followed a 1–3-day delay of positive cases. This delay was not compatible with a biological course of Covid-19 but was compatible with a health management explanation. The second finding is that the Italian number of deaths did not increase linearly with the number of positive cases, but their relationship could be modeled by a second-order polynomial function. The high number of positive cases might have a direct and an indirect effect on the number of deaths, the latter being related to the overwhelmed bed capacity of intensive care units.


title: There is a relationship between obesity and COVID‐19 but more information is needed

text: We read with particular interest your comments in Obesity regarding the Coronavirus‐19 (COVID‐19) epidemic (1) and a related manuscript by Simonnet et al. (2). Resolution of the specific relationship between obesity and COVID‐19, two existing public health epidemics, is critically needed to potentially prevent health systems worldwide from being overburdened. Few studies describing COVID‐19 with rates of obesity exist, and most are based on heterogeneous populations (Table 1) (2‐7). In cohort studies with COVID‐19 disease, obesity rates are generally reported as no higher than population‐based estimates; in contrast, subgroups of critically ill patients (e.g, intensive care unit (ICU)) report higher prevalence rates of obesity.


title: Isolation of patients in psychiatric hospitals in the context of the COVID-19 pandemic: An ethical, legal, and practical challenge

text: Abstract Psychiatric inpatients are particularly vulnerable to the transmission and effects of COVID-19. As such, healthcare providers should implement measures to prevent its spread within mental health units, including adequate testing, cohorting, and in some cases, the isolation of patients. Respiratory isolation imposes a significant limitation on an individual's right to liberty, and should be accompanied by appropriate legal safeguards. This paper explores the implications of respiratory isolation in English law, considering the applicability of the common law doctrine of necessity, the Mental Capacity Act 2005, the Mental Health Act 1983, and public health legislation. We then interrogate the practicality of currently available approaches by applying them to a series of hypothetical cases. There are currently no ‘neat’ or practicable solutions to the problem of lawfully isolating patients on mental health units, and we discuss the myriad issues with both mental health and public health law approaches to the problem. We conclude by making some suggestions to policymakers.


docid: ceuppvgs

title: A skull in a skull: a child’s observation


title: The European race of Gremmeniella abietina hosts a single species of Gammapartitivirus showing a global distribution and possible recombinant events in its history

text: Abstract The population genetics of the family Partitiviridae was studied within the European race of the conifer pathogen Gremmeniella abietina. One hundred sixty-two isolates were collected from different countries, including Canada, the Czech Republic, Finland, Italy, Montenegro, Serbia, Spain, Switzerland, Turkey and the United States. A unique species of G. abietina RNA virus–MS1 (GaRV-MS1) appears to occur indistinctly in G. abietina biotypes A and B, without a particular geographical distribution pattern. Forty-six isolates were shown to host GaRV-MS1 according to direct specific RT-PCR screening, and the virus was more common in biotype A than B. Phylogenetic analysis based on 46 partial coat protein (CP) cDNA sequences divided the GaRV-MS1 population into two closely related clades, while RNA-dependent RNA polymerase (RdRp) sequences revealed only one clade. The evolution of the virus appears to mainly occur through purifying selection but also through recombination. Recombination events were detected within alignments of the three complete CP and RdRp sequences of GaRV-MS1. This is the first time that recombination events have been directly identified in fungal partitiviruses and in G. abietina in particular. The results suggest that the population dynamics of GaRV-MS1 do not have a direct impact on the genetic structure of its host, G. abietina, though they might have had an innocuous ancestral relationship.


An Integrated and Intergenerational Community Response to Promote Holistic Wellbeing During the COVID-19 Pandemic


title: Binding and Repressive Activities of Apolipoprotein E3 and E4 Isoforms on the Human ApoD Promoter

text: Apolipoprotein D (ApoD) gene expression is increased in several neurological disorders such as Alzheimer’s disease (AD) and multiple sclerosis. We previously showed that transgenic mice that overexpress human ApoD show a better resistance against paraquat or OC43 coronavirus-induced neurodegeneration. Here, we identified several nuclear factors from the cortex of control and OC43-infected mice which bind a fragment of the proximal ApoD promoter in vitro. Of interest, we detected apolipoprotein E (ApoE). Human ApoE consists of three isoforms (E2, E3, and E4) with the E4 and E2 alleles representing a greater and a lower risk for developping AD, respectively. Our results show that ApoE is located in the nucleus and on the ApoD promoter in human hepatic and glioblastoma cells lines. Furthermore, overexpression of ApoE3 and ApoE4 isoforms but not ApoE2 significantly inhibited the ApoD promoter activity in U87 cells (E3/E3 genotype) cultured under normal or different stress conditions while ApoE knock-down by siRNA had a converse effect. Consistent with these results, we also demonstrated by ChIP assay that E3 and E4 isoforms, but not E2, bind the ApoD promoter. Moreover, using the Allen Brain Atlas in situ hybridization database, we observed an inverse correlation between ApoD and ApoE mRNA expression during development and in several regions of the mouse brain, notably in the cortex, hippocampus, plexus choroid, and cerebellum. This negative correlation was also observed for cortex layers IV–VI based on a new Transcriptomic Atlas of the Mouse Neocortical Layers. These findings reveal a new function for ApoE by regulating ApoD gene expression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12035-013-8456-0) contains supplementary material, which is available to authorized users.


title: Affective temperament, attachment style, and the psychological impact of the COVID-19 outbreak: an early report on the Italian general population

text: Abstract The outbreak of COVID-19 is severely affecting mental health worldwide, although individual response may vary. This study aims to investigate the psychological distress perceived by the Italian general population during the early phase of the COVID-19 pandemic, and to analyze affective temperament and adult attachment styles as potential mediators. Through an online survey, we collected sociodemographic and lockdown-related information and evaluated distress, temperament, and attachment using the Kessler 10 Psychological Distress Scale (K10), the Temperament Evaluation of Memphis, Pisa, Paris and San Diego-Autoquestionnaire short version (TEMPS-A) and the Attachment Style Questionnaire (ASQ). In our sample (n=500), 62% of the individuals reported no likelihood of psychological distress, whereas 19.4% and 18.6% displayed mild and moderate-to-severe likelihood. Cyclothymic (OR: 1.24; p<0.001), depressive (OR: 1.52; p<0.001) and anxious (OR: 1.58; p=0.002) temperaments, and the ASQ “Need for approval” (OR: 1.08; p=0.01) were risk factors for moderate-to-severe psychological distress compared to no distress, while the ASQ “Confidence” (OR: 0.89; p=0.002) and “Discomfort with closeness” were protective (OR: 0.92; p=0.001). Cyclothymic (OR: 1.17; p=0.008) and depressive (OR: 1.32; p=0.003) temperaments resulted as risk factors in subjects with moderate-to-severe psychological distress compared to mild distress, while the ASQ “Confidence” (OR: 0.92; p=0.039) and “Discomfort with closeness” (OR: 0.94; p=0.023) were protective. Our data indicated that a relevant rate of individuals may have experienced psychological distress following the COVID-19 outbreak. Specific affective temperament and attachment features predict the extent of mental health burden. To the best of our knowledge, these are the first data available on the psychological impact of the early phase of the COVID-19 pandemic on a sizeable sample of the Italian population. Moreover, our study is the first to investigate temperament and attachment characteristics in the psychological response to the ongoing pandemic. Our results provide further insight into developing targeted intervention strategies.


Behavioral and psychosocial responses of people receiving treatment for advanced lung cancer during the COVID‐19 pandemic: a qualitative analysis


Use of Biphasic Cuirass Ventilation During Bronchoscopy in a Critically Ill Infant


title: Immunostimulatory and anti-metastatic activity of polysaccharides isolated from byproducts of the corn starch industry

text: Abstract Corn steep liquor (CSL) is a major by-product of the corn steeping process that is utilized in the wet milling industry. To develop new physiologically active polysaccharides from CSL, polysaccharides were isolated and their innate immunostimulatory and anti-metastatic activities were investigated. Corn byproduct polysaccharides (CBP) were preferentially isolated from CSL and further separated into supernatant (CBP1S) and precipitate (CBP1P) fractions. The anti-complementary activity of CBP1S was more potent than CBP1P and CBP. In addition, CBP1S enhanced production of macrophage-stimulating cytokines (e.g., IL-6 and IL-12) and natural killer (NK) cell-activating substances (e.g., granzyme and interferon-γ). Further, CBP1S significantly inhibited lung metastasis at a dose of 1000μg per mouse in an experimental lung metastasis model. These results suggest that CBP1S seems to promote the inhibition of lung metastasis through a mechanism leading to stimulation of the innate immune system, and CBP1S could be used as immunostimulating agents and for industrial applications.


title: The easy way is hard enuff

text: Andrew Murray argues that you can use your forced exile from the lab to produce better future experiments by dissecting your past failures and successes and collaboratively critiquing the experiments you’re planning for your return to the lab.


docid: hypxhbuu

title: A novel biosensor based on serum antibody immobilization for rapid detection of viral antigens

text: Abstract In this paper, we represent a label-free biosensor based on immobilization of serum antibodies for rapid detection of viral antigens. Human serum containing specific antibodies against Japanese encephalitis virus (JEV) was immobilized on a silanized surface of an interdigitated sensor via protein A/glutaraldehyde for electrical detection of JEV antigens. The effective immobilization of serum antibodies on the sensor surface was verified by Fourier transform infrared spectrometry and fluorescence microscopy. The signal of the biosensor obtained by the differential voltage converted from the change into non-Faradic impedance resulting from the specific binding of JEV antigens on the surface of the sensor. The detection analyzed indicates that the detection range of this biosensor is 1–10μg/ml JEV antigens, with a detection limit of 0.75μg/ml and that stable signals are measured in about 20min. This study presents a useful biosensor with a high selectivity for rapid and simple detection of JEV antigens, and it also proposes the biosensor as a future diagnostic tool for rapid and direct detection of viral antigens in clinical samples for preliminary pathogenic screenings in the case of possible outbreaks.


title: Association of COVID-19 Infections in San Francisco in Early March 2020 with Travel to New York and Europe

text: Real-time dissemination of epidemiological survey data from positive COVID-19 cases is critical to support efforts to contain or reduce spread of viral infection in the community. Here we detected a significant association between domestic travel or travel to Europe and the identification of new cases in San Francisco, California, USA. These findings suggest that domestic and European travelers may need to be prioritized for evaluation of acute infection from COVID-19 in the setting of limited testing capacity.


title: Evolutionary Relationships between Bat Coronaviruses and Their Hosts

text: Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness in humans during 2002–2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.


1 year after The Lancet Neonatal Survival Series—was the call for action heard?


title: Is 0.5% Hydrogen Peroxide Effective against SARS‐CoV‐2?

text: SARS‐CoV‐2 is mostly transmitted through respiratory droplets and contact routes, but the WHO also states that “airborne transmission may be possible in specific circumstances and settings in which procedures or support treatments that generate aerosols are performed” (WHO, 2020).


A child presenting with severe hypertension and circulatory failure—a diagnostic conundrum: Answers


title: Automated detection of COVID-19 cases using deep neural networks with X-ray images

text: Abstract The novel coronavirus 2019 (COVID-2019), which first appeared in Wuhan city of China in December 2019, spread rapidly around the world and became a pandemic. It has caused a devastating effect on both daily lives, public health, and the global economy. It is critical to detect the positive cases as early as possible so as to prevent the further spread of this epidemic and to quickly treat affected patients. The need for auxiliary diagnostic tools has increased as there are no accurate automated toolkits available. Recent findings obtained using radiology imaging techniques suggest that such images contain salient information about the COVID-19 virus. Application of advanced artificial intelligence (AI) techniques coupled with radiological imaging can be helpful for the accurate detection of this disease, and can also be assistive to overcome the problem of a lack of specialized physicians in remote villages. In this study, a new model for automatic COVID-19 detection using raw chest X-ray images is presented. The proposed model is developed to provide accurate diagnostics for binary classification (COVID vs. No-Findings) and multi-class classification (COVID vs. No-Findings vs. Pneumonia). Our model produced a classification accuracy of 98.08% for binary classes and 87.02% for multi-class cases. The DarkNet model was used in our study as a classifier for the you only look once (YOLO) real time object detection system. We implemented 17 convolutional layers and introduced different filtering on each layer. Our model (available at (https://github.com/muhammedtalo/COVID-19)) can be employed to assist radiologists in validating their initial screening, and can also be employed via cloud to immediately screen patients.


title: Children's (and autoimmune patients) morbidity (and mortality) from Covid‐19 is similar to the general population: immunologic rationale.

text: We read with great interest the Viewpoint by L. H Henderson et al (1) on the therapeutic approach with Glucocorticoids (GC) to the inflammation and Cytokine Storm phases of SARS.CoV 2 infection. We would like to expand their analysis and discuss the data , so far reported in Children and Autoimmune patients ( Rheumatoid Arthritis , Systemic Lupus Erythematosus), about the chance of undergoing a "severe" infection. So far children (and autoimmune patients) , who should be extremely fragile, rarely entered into the third phase "the cytokine release syndrome‐CRS " of COVID‐19, leading only some patients to the Intensive Care Units (ICUs).


title: Severe acute respiratory syndrome: identification of the etiological agent

text: Abstract The severe acute respiratory syndrome (SARS) emerged in late 2002 in southern China and rapidly spread to countries around the globe. Three research groups within a World Health Organization (WHO)-coordinated network have independently and simultaneously shown that a novel coronavirus is linked to SARS. A fourth group has completed the Koch's postulates by infecting monkeys with the agent. Sequencing of the complete genome was achieved only weeks after the first isolate of the virus became available.


title: La gestión de la asistencia neurológica en tiempos de la pandemia de COVID-19

text: Abstract Introducción: La pandemia de Covid-19 ha supuesto la necesidad de tomar de decisiones para mantener la asistencia neurológica, sin precedentes. En este artículo se analizan esa toma de decisiones operativas. Desarrollo: Los autores refieren las fórmulas empleadas como son la realización de un plan de reorganización funcional, estrategias para la hospitalización y urgencias, la realización de consultas telefónicas para el mantenimiento de la atención neurológica, la asistencia en una área externa al hospital para pacientes preferentes, las decisiones sobre exploraciones complementarias y tratamientos intrahospitalarios periódicos o implantar un teléfono para la priorización de crisis epilépticas. Conclusión: A pesar de la situación de aislamiento, los servicios de neurología deben mantener la atención de sus pacientes, a través de distintas fórmulas operativas, que como cualquier elemento de gestión, deberán evaluarse. Abstract Introduction: The COVID-19 epidemic has led to the need for unprecedented decisions to be made to maintain the provision of neurological care. This article addresses operational decision-making during the epidemic. Development: We report the measures taken, including the preparation of a functional reorganisation plan, strategies for hospitalisation and emergency management, the use of telephone consultations to maintain neurological care, provision of care at a unit outside the hospital for priority patients, decisions about complementary testing and periodic in-hospital treatments, and the use of a specific telephone service to prioritise patients with epileptic seizures. Conclusion: Despite the situation of confinement, neurology departments must continue to provide patient care through different means of operation. Like all elements of management, these must be evaluated.


title: Age-related increase of porcine natural interferon α producing cell frequency and of interferon yield per cell

text: Abstract Porcine blood mononuclear cells (PBMC) were shown to secrete interferon α (IFN-α) after induction by a coronavirus, the transmissible gastroenteritis virus (TGEV). IFN-α producing cells, referred to as natural interferon α producing (NIP) cells, were detected by an ELISPOT assay using anti-porcine IFN-α monoclonal antibodies. The frequency of NIP cells among blood cells is low, at most 40–110 per 105 PBMC and each NIP cell was found to produce several units of IFN. We have shown that NIP cell frequency and IFN yield per cell gradually increased with the age of the donor animals, from the neonatal period to the adult age, with a significant increase around puberty. Our present results also indicate that NIP cells may be influenced by physiological and genetic factors; thus (1) NIP cell frequency and IFN yield per cell were decreased during lactation; (2) Chinese (Meishan) pigs were found to have higher NIP cell frequency and IFN yield per cell than European (Large White) animals.


title: A vesicular stomatitis virus-based mucosal vaccine promotes dendritic cell maturation and elicits preferable immune response against coxsackievirus B3 induced viral myocarditis

text: Abstract Recombinant vesicular stomatitis virus (VSV) is widely used as a vaccine platform. However, the capacity of VSV-based vaccines to induce mucosal immunity has not been fully investigated. In the present study, a recombinant VSV expressing coxsackievirus B3 (CVB3) major immunogen VP1 has been generated and the immune protection elicited by VSV-VP1 was evaluated. We demonstrated that intranasal delivery of VSV-VP1 can induce a potent antigen-specific mucosal immune response as well as a systemic immune response, particularly the induction of polyfunctional T cells. Importantly, mice immunized with VSV-VP1 were better protected against CVB3-induced viral myocarditis than those receiving a chitosan-formulated DNA vaccine. Increased dendritic cell (DC) maturation in the mesenteric lymph node (MLN) was observed in the mice vaccinated with VSV-VP1, which could be a potential mechanism for the protective immune response. These findings support VSV as a viral delivery vector that can induce robust mucosal immunity that should be considered for further vaccine development.


title: Therapeutic Algorithm for Use of Melatonin in Patients With COVID-19

text: The coronavirus, COVID-19, has infected hundreds of thousands and killed tens of thousands of individuals worldwide. This highly infectious condition continues to ravage the world population and has yet to reach it peak infective rate in some countries. Many conventional drugs including hydroxychloroquine/chloroquine, lopinavir, remdesivir, etc., have been repurposed as treatments for this often deadly disease, but there is no specifically-designed effective drug available; also, the drugs mentioned have significant side effects and their efficacy is unknown. New drugs and vaccines are being designed as COVID-19 treatment, but their development and testing will require months to years. Time is not a luxury that this crisis has. Thus, there is a serious unmet need for the identification of currently-available and safe molecules which can be used to slow or treat COVID-19 disease. Here, we suggest melatonin be given consideration for prophylactic use or treatment alone or in combination with other drugs. Melatonin's multiple actions as an anti-inflammatory, anti-oxidant, and anti-viral (against other viruses) make it a reasonable choice for use. Melatonin is readily available, can be easily synthesized in large quantities, is inexpensive, has a very high safety profile and can be easily self-administered. Melatonin is endogenously-produced molecule in small amounts with its production diminishing with increased age. Under the current critical conditions, large doses of melatonin alone or in combination with currently-recommended drugs, e.g., hydroxychloroquine/chloroquine, to resist COVID-19 infection would seem judicious.


title: Cumulative pharmacological activity index volumes 1-30

text: Publisher Summary This chapter lists the important subjects on pharmacological activity that are discussed in the publication Studies in Natural Products Chemistry, Volumes 1–30, such as abdominal constriction test, acanthoic acid, acetaminophen, Parkinson's disease, photodynamic activity, prostaglandins, and oleanolic acid. The terms are mentioned along with the page numbers in which they are discussed in the publication.


title: Arginine metabolism during macrophage autocrine activation and infection with mouse hepatitis virus 3

text: Abstract In contrast to BALB/c mouse macrophages (MΦ), MΦ from the A/J mouse strain, upon activation by exogenous interferon gamma (IFNγ), develop an anti-mouse hepatitis virus 3 (MHV3) state which correlates with resistance to virus infection. To investigate the autocrine activation of BALB/c and A/J MΦ, we activated them with interleukin-12 (IL-12) and/or IL-18, and quantified IFNγ production, the anti-MHV3 state and arginine metabolism. Synergistic activation by IL-12/IL-18 induced the expression of the IFNγ gene in MΦ from both mouse strains. In bone marrow (BM) or peritoneal (P) MΦ of specific pathogen-free (spf) mice of both strains, IFNγ synthesis occurred only with a synergistic IL-12/IL-18 activation and showed increasing levels from 24 to 72h of activation. In contrast, when non-spf mice were used in the assay, their PMΦ synthesized higher IFNγ levels upon activation with only IL-12 or only IL-18 or both. The BALB/c MΦ were always capable of synthesizing higher amounts of IFNγ than the A/J MΦ. An anti-MHV3 state was observed only in A/J MΦ upon activation with IL-12/IL-18 or IFNγ regardless of their origin from the peritoneum or bone marrow. Arginine metabolism in activated and/or virus infected BMMΦ was investigated through nitric oxide (NO) and arginase induction as well as the consumption of arginine and synthesis of citrulline, ornithine and spermine. The results showed that both BALB/c and A/J BMMΦ populations released NO only after activation with IL-12/IL-18 or IFNγ. Arginase was not induced in BMMΦ from both strains by IL-12/IL-18 or IFNγ but only by IL-4/IL-10. Higher arginine consumption was observed in BMMΦ from both strains upon activation with IL-4 or IFNγ which further increased, in this case, when the cells were infected with MHV3. As a consequence of nitric oxide synthase synthesis and arginine consumption in IFNγ activated BMMΦ, we observed a higher synthesis of citrulline. High levels of ornithine were induced only upon IL-4 activation. Polyamine synthesis was higher in A/J BMMΦ than in BALB/c ones, which correlated with the slightly lower levels of ornithine observed. Upon infection with MHV3, we observed a higher synthesis of spermine. IL-12/IL-18 or IFNγ activation, mainly in MHV3 infected cells, led to a decreased synthesis of polyamines, notably spermine, only in A/J BMMΦ. Difluoromethylornithine treatment, which leads to inhibition of polyamine synthesis, induced a decreased MHV3 multiplication in both BALB/c and A/J BMMΦ. Altogether these data show the relevance of IFNγ, from the autocrine or paracrine pathway, and arginine metabolism for the control of MHV3 replication in MΦ of a resistant mouse strain.


title: Two highly similar LAEDDTNAQKT and LTDKIGTEI epitopes in G glycoprotein may be useful for effective epitope based vaccine design against pathogenic Henipavirus

text: Abstract Nipah virus and Hendra virus, two members of the genus Henipavirus, are newly emerging zoonotic pathogens which cause acute respiratory illness and severe encephalitis in human. Lack of the effective antiviral therapy endorses the urgency for the development of vaccine against these deadly viruses. In this study, we employed various computational approaches to identify epitopes which has the potential for vaccine development. By analyzing the immune parameters of the conserved sequences of G glycoprotein using various databases and bioinformatics tools, we identified two potential epitopes which may be used as peptide vaccines. Using different B cell epitope prediction servers, four highly similar B cell epitopes were identified. Immunoinformatics analyses revealed that LAEDDTNAQKT is a highly flexible and accessible B-cell epitope to antibody. Highly similar putative CTL epitopes were analyzed for their binding with the HLA-C 12*03 molecule. Docking simulation assay revealed that LTDKIGTEI has significantly lower binding energy, which bolstered its potential as epitope-based vaccine design. Finally, cytotoxicity analysis has also justified their potential as promising epitope-based vaccine candidate. In sum, our computational analysis indicates that either LAEDDTNAQKT or LTDKIGTEI epitope holds a promise for the development of universal vaccine against all kinds of pathogenic Henipavirus. Further in vivo and in vitro studies are necessary to validate the obtained findings.


title: Intestinal Organoids—Current and Future Applications

text: Recent technical advances in the stem cell field have enabled the in vitro generation of complex structures resembling whole organs termed organoids. Most of these approaches employ culture systems that allow stem cell-derived or tissue progenitor cells to self-organize into three-dimensional (3D)-structures. Since organoids can be grown from different species (human, mouse, cat, dog), organs (intestine, kidney, brain, liver), and from patient-derived induced pluripotent stem cells, they create significant prospects for modelling development and diseases, for toxicology and drug discovery studies, and in the field of regenerative medicine. Here, we report on intestinal stem cells, organoid culture, organoid disease modeling, transplantation, specifically covering the current and future uses of this exciting new insight model to the field of veterinary medicine.


title: Broad-spectrum microbicidal activity, toxicologic assessment, and materials compatibility of a new generation of accelerated hydrogen peroxide-based environmental surface disinfectant

text: BACKGROUND: Concerns on human and environmental safety and label claims of many microbicides point to the need for safer, faster acting, and broad-spectrum substitutes. ACCEL TB, a 0.5% accelerated H(2)O(2) (AHP)-based disinfectant described here, is a potential candidate. METHODS: ACCEL TB was tested for its broad-spectrum microbicidal activity, safety and materials compatibility using internationally accepted protocols. Activity against bacteria (Staphylococcus aureus, Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa, vancomycin-resistant Enterococcus faecalis, methicillin-resistant Staphylococcus aureus, and Salmonella choleraesuis) was tested with the AOAC use-dilution method and the first tier of a quantitative carrier test (QCT-1). Mycobactericidal activity was tested against Mycobacterium bovis and Mycobacterium terrae using a quantitative suspension test (QST) and QCT-1, respectively. Fungicidal activity (Trichophyton mentagrophytes) was determined with the AOAC test and QCT-1. Activity against several enveloped and nonenveloped viruses was evaluated using the American Society for Testing and Materials (ASTM) method No. E-1053. Sanitizing action was tested against 7 types of vegetative bacteria with method No. DIS/TSS-10. All microbicidal tests contained an added soil load; in all AOAC tests, it was 5% fetal bovine serum, and, in QCT-1, a mixture of 3 types of proteins in phosphate buffer was used instead. The methods to test for acute oral, dermal, inhalation toxicities, and dermal and eye irritation as well as skin sensitization complied with the requirements of the Organization for Economic Cooperation and Development and the US Environmental Protection Agency (OPPTS 870). Standard methods were also used to test compatibility with metals and plastics. RESULTS: At 20°C, the full-strength product was bactericidal and virucidal in 1 minute and mycobactericidal and fungicidal in 5 minutes. It was nonirritating to skin and eyes. The acute oral LD(50) (lethal dose 50%) was >5000 mg/kg. It was compatible with 12 types of plastic and 3 out of 4 metals. CONCLUSION: The tested formulation showed a high safety and materials compatibility profile in addition to being a fast acting, intermediate-level disinfectant.


title: Molecular Evolution of the Primate α-/θ-Defensin Multigene Family

text: The primate α-/θ-defensin multigene family encodes versatile endogenous cationic and amphipathic peptides that have broad-spectrum antibacterial, antifungal and antiviral activity. Although previous studies have reported that α-/θ-defensin (DEFA/DEFT) genes are under birth-and-death evolution with frequent duplication and rapid evolution, the phylogenetic relationships of the primate DEFA/DEFT genes; the genetic bases for the existence of similar antimicrobial spectra among closely related species; and the evolutionary processes involved in the emergence of cyclic θ-defensins in Old World monkeys and their subsequent loss of function in humans, chimpanzees and gorillas require further investigation. In this study, the DEFA/DEFT gene repertoires from primate and treeshrew were collected, followed by detailed phylogenetic, sequence and structure, selection pressure and comparative genomics analyses. All treeshrew, prosimian and simian DEFA/DEFT genes are grouped into two major clades, which are tissue-specific for enteric and myeloid defensins in simians. The simian enteric and myeloid α-defensins are classified into six functional gene clusters with diverged sequences, variable structures, altered functional constraints and different selection pressures, which likely reflect the antimicrobial spectra among closely related species. Species-specific duplication or pseudogenization within each simian cluster implies that the antimicrobial spectrum is ever-shifting, most likely challenged by the ever-changing pathogen environment. The DEFT evolved from the myeloid DEFA8. The prosegment of θ-defensin is detected with adaptive changes coevolving with the new protein fold of mature peptide, coincident with the importance of the prosegment for the correct folding of the mature peptide. Lastly, a less-is-hitchhiking hypothesis was proposed as a possible explanation for the expansion of pseudogene DEFTP and the loss of functional DEFT, where the gain or loss of the hitchhiker is determined by its adjacent driver gene during the birth-and-death evolutionary process.


title: Adenoviruses as vectors for delivering vaccines to mucosal surfaces

text: Abstract Immunization of mucosal surfaces has become an attractive route of vaccine delivery because of its ability to induce mucosal immunity. Although various methods of inducing mucosal immunity are being developed, our laboratory has focused on developing adenoviruses as replication–competent and replication–incompetent vectors. The present report will summarize our progress in sequencing the entire bovine adenovirus-3 genome and identifying regions which can be deleted and subsequently used as insertion sites for foreign genes in developing recombinant viral vaccines. Using these recombinant viruses, we demonstrated the ‘proof-of-principle’ in developing mucosal immunity and, more importantly, inducing protection against bovine herpes virus in a natural host–cattle. Finally, we demonstrated that immunity and protection occurred even in animals that had pre-existing antibodies to the vector.


title: Immunogenicity of a DNA-launched replicon-based canine parvovirus DNA vaccine expressing VP2 antigen in dogs

text: Abstract A replicon-based DNA vaccine encoding VP2 gene of canine parvovirus (CPV) was developed by cloning CPV-VP2 gene into a replicon-based DNA vaccine vector (pAlpha). The characteristics of a replicon-based DNA vaccine like, self-amplification of transcripts and induction of apoptosis were analyzed in transfected mammalian cells. When the pAlpha-CPV-VP2 was injected intradermal as DNA-launched replicon-based DNA vaccine in dogs, it induced CPV-specific humoral and cell mediated immune responses. The virus neutralization antibody and lymphocyte proliferative responses were higher than conventional CPV DNA vaccine and commercial CPV vaccine. These results indicated that DNA-launched replicon-based CPV DNA vaccine was effective in inducing both CPV-specific humoral and cellular immune responses and can be considered as effective alternative to conventional CPV DNA vaccine and commercial CPV vaccine.


Role of chest CT in patients with acute abdomen during the COVID‐19 era


title: Increase in the prescription rate of antidepressants after the Sewol Ferry disaster in Ansan, South Korea

text: Abstract Background Previous pharmaco-epidemiological studies have reported increases in the prescription of psychotropic medications after a disaster, reflecting post-disaster changes in psychiatric conditions and mental health service utilization. We investigated changes in the prescription of psychotropic medications in the Danwon district of Ansan city (Ansan Danwon) compared to a control community before and after the Sewol Ferry disaster on April 16, 2014. Methods Data was collected from the Korean Health Insurance Review and Assessment Service database. We analyzed the prescription rates of psychotropic medications including antidepressants, anxiolytics, and sedatives/hypnotics, and investigated whether the time-series pattern of monthly prescriptions per 100,000 people was different in Ansan Danwon compared to that in Cheonan city after the Sewol Ferry disaster through difference-in-differences regression analysis. Results Ansan Danwon showed a significantly greater increase (5.6%) in the prescription rate of antidepressants compared to Cheonan city following the Sewol Ferry disaster. There were no significant differences in changes in the prescription rates of anxiolytics or sedatives/hypnotics. In the secondary analysis, a significantly greater increase in the prescription rate of antipsychotics was observed in Ansan Danwon compared to a control community after the disaster. Limitations We could not exclude the possibility that other events influenced changes in the prescription rates of psychotropic medications during the study period. Conclusions Pharmaco-epidemiological studies on psychotropic medication prescription after a disaster provide important information about population-level mental health. Our results suggest that the Sewol Ferry disaster exerted a harmful effect on the mental health status of the affected community.


title: Degenerate primers based RT-PCR for rapid detection and differentiation of airborne chicken Newcastle disease virus in chicken houses

text: Abstract Airborne Newcastle disease (ND) viruses in the air of five chicken houses were detected and differentiated by reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate primers. Fifteen air samples were collected with All Glass Impinger-30 (AGI-30) air samplers in each house. Airborne ND viruses were also isolated and virulence identified by in vivo tests. Avirulent viruses were detected both in air samples and swab samples in four houses by degenerate primers based RT-PCR. Virulent viruses were detected only in the air samples by degenerate primers based RT-PCR in two houses. Seven strains viruses were isolated from the RT-PCR positive air samples. Of the seven strains, three strains were virulent viruses and four strains were avirulent viruses identified by in vivo tests. The results showed that it was feasible to detect and differentiate NDV in the air samples using degenerate primers based RT-PCR. This technique could decrease the time it required identify NDV infected flocks while distinguishing between virulent and avirulent viruses. It will help effectively to control Newcastle disease.


title: Optimization of extraction and PCR amplification of RNA extracts from paraffin-embedded tissue in different fixatives

text: Abstract A method was developed for fast and efficient isolation of RNA from paraffin-embedded tissue sections for subsequent PCR analysis. This method is based on the binding of RNA to acid-treated glass beads in the presence of a high molarity of guanidinium salt. It can be completed within an hour, and obviates the need for dewaxing and phenol/chloroform extractions. The effect of various fixatives and fixation times was tested and the amplification of actin mRNA fragments ranging in length from 82 to 507 bp was used to demonstrate the presence of RNA in the extracts. The method was compared to existing extraction techniques by studying the quality of the templates for reverse-transcriptase polymerase chain reaction amplification (RT-PCR), using virus-infected and mock-infected paraffin-embedded cell pellets as a model. PCR amplification of cellular and viral RNA was successful for RNA isolated by use of all extraction techniques, although the glass bead method was preferred for its simplicity and rapidity. Specimens fixed with formalin were found to be suitable for PCR, but the best results were obtained with acetone-fixed paraffin-embedded material. Dewaxing of tissue sections had no effect on the yield and quality of RNA extractions, and further purification of the extracts using gel filtration did not improve the results. After the protocols were optimized, rotavirus-infected cell pellets were used to demonstrate that extraction and amplification of dsRNA was possible. The information obtained from the studies with the model system was used for extraction of toroviral and rotaviral RNA from archival intestinal material. These data indicate that paraffin-embedded archival tissue can be used for RT-PCR analysis, adding an important technique to diagnostic pathology and retrospective studies.


title: De quoi le Covid-19 est-il le nom?

text: Résumé L’irruption massive de l’expression Covid-19 dans le milieu hospitalier comme dans nos vies a fait apparaitre une étrange analogie entre l’expansion d’un virus et la dissémination virale, elle aussi, d’une manière de parler, de formuler et de nommer pour agir. Cet article se propose de questionner l’impact de l’expression Covid-19 sur le langage des soignants. Il propose de discuter la dimension éthique d’un soin des mots du soin lorsqu’une manière de parler s’impose à eux. Il interroge également le pluralisme épistémologique et éthique engageant la médecine entre science, clinique et politique. Abstract The massive irruption of Covid-19 expression in the hospital environment as well as in our lives has brought to light a strange analogy between the expansion of a virus and viral dissemination, too, in a way of speaking and naming for action. This article proposes to question the impact of the expression Covid-19 on the language of caregivers. It proposes to discuss the ethical dimension of a care of care words when a way of speaking imposes itself on them. It also questions the epistemological and ethical pluralism involving medicine between science, clinical and politics.


title: Does autoimmunity of endogenous vasoactive neuropeptides cause retinopathy in humans?

text: Summary Immune privilege is a physiologic mechanism within the eye which protects it against pathogens, while also protecting it from inflammation. Immunological mechanisms in the eye must be tightly regulated to ensure externally mediated injury and infection or internally mediated autoimmunity do not exceed self-defence tolerance. Vasoactive neuropeptides (VNs) including vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase polypeptide (PACAP) and their receptors exist in the mammalian eye including the sclera, cornea, iris, ciliary body, ciliary process and the retina and may have a role in protecting these normally immune privileged sites. VN receptors are class II G protein-coupled receptors (GPCRs) which couple primarily to the adenylate cyclase (AC)-cyclic AMP pathway. A sound blood supply is essential for retinal survival hence vascular compromise will have serious consequences. Retinal vasculitis is a potentially blinding condition with a strong association with systemic inflammatory diseases. Compromise of the endothelial barriers and the blood retina barrier (BRB) may instigate inflammatory responses setting up a chain of events involving VNs in a manner which provokes autoimmunity to them. Protection from BRB breakdown may be linked to nitric oxide (NO) effects and actions of phosphodiesterase inhibitors and cAMP production. Induced NO expressed under influences of inflammatory mediators evokes neurodegeneration and cell apoptosis and may lead to serious ocular disease including retinal injury. Other inflammatory mediators also play a role in retinal pathology. PACAP and glutamate are co-stored in the retinohypothalamic tract and PACAP attenuates glutamate induced neurotoxicity in cultured retinal neurons suggesting that compromise of this VN would have significant detrimental impact on retinal viability though glutamate toxicity. Additional effects of VN compromise would possibly occur through unopposed vasoconstriction and inflammation. Proof of this hypothesis has important implications for treatment and prevention of autoimmune retinopathy and blindness as a number of therapeutic pathways may be opened. Importantly for therapeutic contexts cAMP effects are maintained by phosphodiesterase (PDE) inhibitors which could be used in VN autoimmune disorders. A compelling case may exist to undertake a therapeutic trial of VN replacement, PDE inhibitors and other agents in autoimmune retinopathies resulting from possible VN autoimmunity.


Beyond COVID‐19 lockdown: A Coasean approach with optionality


title: Removal of chronic Mycoplasma ovipneumoniae carrier ewes eliminates pneumonia in a bighorn sheep population

text: 1. Chronic pathogen carriage is one mechanism that allows diseases to persist in populations. We hypothesized that persistent or recurrent pneumonia in bighorn sheep (Ovis canadensis) populations may be caused by chronic carriers of Mycoplasma ovipneumoniae (Mo). Our experimental approach allowed us to address a conservation need while investigating the role of chronic carriage in disease persistence. 2. We tested our hypothesis in two bighorn sheep populations in South Dakota, USA. We identified and removed Mo chronic carriers from the Custer State Park (treatment) population. Simultaneously, we identified carriers but did not remove them from the Rapid City population (control). We predicted removal would result in decreased pneumonia, mortality, and Mo prevalence. Both population ranges had similar habitat and predator communities but were sufficiently isolated to preclude intermixing. 3. We classified chronic carriers as adults that consistently tested positive for Mo carriage over a 20‐month sampling period (n = 2 in the treatment population; n = 2 in control population). 4. We failed to detect Mo or pneumonia in the treatment population after chronic carrier removal, while both remained in the control. Mortality hazard for lambs was reduced by 72% in the treatment population relative to the control (CI = 36%, 91%). There was also a 41% reduction in adult mortality hazard attributable to the treatment, although this was not statistically significant (CI = 82% reduction, 34% increase). 5. Synthesis and Applications: These results support the hypothesis that Mo is a primary causative agent of persistent or recurrent respiratory disease in bighorn sheep populations and can be maintained by a few chronic carriers. Our findings provide direction for future research and management actions aimed at controlling pneumonia in wild sheep and may apply to other diseases.


title: Characterization of BIV Env core: Implication for mechanism of BIV-mediated cell fusion

text: Abstract Entry of lentiviruses, such as human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV), requires folding of two heptad repeat regions (HR1 and HR2) of gp41 into a trimer-of-hairpins, which subsequently brings virus and cell membrane into fusion. This motif is a generalized feature of viral fusion proteins and has been exploited in generating antiviral fusion agents. In the present paper, we report structural characters of Env protein from another lentivirus, bovine immunodeficiency virus (BIV), which contributes to a good animal model of HIV. BIV HR1 and HR2 regions are predicted by two different programs and expressed separately or conjointly in Escherichia coli. Biochemical and biophysical analyses show that the predicted HRs of BIV Env can form a stable trimer-of-hairpins or six-helix bundle just like that formed by feline immunodeficiency virus Env. Cell fusion assay demonstrates that the HR2 peptide of BIV can efficiently inhibit the virus-mediated cell fusion.


title: Knowledge and attitude of healthcare workers and patients on healthcare associated infections in a regional hospital in Ghana

text: OBJECTIVE: To assess knowledge and attitude of healthcare workers (HCWs) and patients on healthcare associated infections (HAIs) in the central regional hospital in Ghana. METHODS: The purposive random sampling method was used to administer questionnaires over a period of 6 months to HCWs and patients visiting the hospital. RESULTS: A total of 210 patients and 71 HCWs were sampled. One hundred and three (53.8%) patients had some knowledge of HAIs with 52 (28.4%) being informed by a HCW compared with 63 (88.7%) of HCWs who were well informed about HAIs. Ninety-seven (46.2%) responding patient always washed their hands while 65 (31%) and 48 (22.9%) respectively sometimes or never washed their hands within or after leaving the hospital. Out of those who washed their hands, 64 (39.5%) always washed with soap while 46 (28.4%) did sometimes. This positively and significantly correlated (r=0.440, P<0.001) with knowledge on HAIs which was however insignificant in HCWs (r=0.025, P=0.835). As many as 48 (67.6%) of HCWs believed that authorities in the hospital had done little to prevent HAIs with the main reason being that the hospital was unclean. Whereas, 112 (53.3%) of patients considered the hospital clean. Twenty-seven (38%) of HCWs had had confirmed HAIs of which cholera made up 12 (16.9%) while 94 (44.8%) of patients believed they had had unconfirmed HAIs. CONCLUSIONS: Although knowledge on HAIs is adequate, low compliance on preventive techniques resulting in high HAIs indicates attitudinal change is the best means of prevention.


title: The Never‐ending Shift: A feminist reflection on living and organizing academic lives during the coronavirus pandemic

text: This article offers a feminist reflection written as a nocturnal stream of consciousness exposing the embodied, emotional and professional experience of living and working during a pandemic outbreak. Framed within a feminist approach, this personal narrative provides an example of the effects of such unexpected and unprecedented circumstances on personal and professional academic lives. Developed during the first stage of the (inter)national coronavirus pandemic, my reflections address issues of privilege; emotional labour; the virtual invasion of the home space within the current increasingly ambiguous space of ‘the workplace'; workload; and wellbeing. Further, I consider how the newly enforced flexible work measures based on online tools have turned current work–life dynamics into a ‘Never‐ending Shift'.


title: Respiratory infections associated with enterovirus D68 from 2011 to 2015 in Beijing, China

text: Enterovirus D68 (EV‐D68) is an emergent viral pathogen associated with mild to severe respiratory infections. In this study, we describe respiratory infections associated with EV‐D68 in Beijing over a 4 year period. Total nucleic acid was extracted from 7,945 clinical specimens collected between January 5, 2011 and July 30, 2015 in Beijing and used for detecting EV‐D68 and other enteroviruses by real‐time PCR. Overall, 555/7,945 (6.99%) specimens were enterovirus positive: 12/7,945 (0.2%) specimens were EV‐D68 positive. Of these patients, 11 were pediatric patients and 1 was a 76‐year‐old man. The main symptoms for the 12 EV‐D68 positive patients were fever (10/12, 83.3%) and cough (6/12, 50%). Ten EV‐D68 infection cases were identified in autumn or winter season. The phylogenetic relationships of the 12 EV‐D68 viral strains with other strains were analyzed based on the sequences of viral protein 1(VP1). The EV‐D68 strains from 2011 to 2013 belonged to groups 1 or 3, while all strains in 2014 were clustered into group 1 together with the strains circulating in the USA. In conclusion, EV‐D68 played a role in respiratory infections in Beijing during this period. In addition, the most common EV‐D68 strain detected was similar to that circulating in the USA in 2014. J. Med. Virol. 88:1529–1534, 2016. © 2016 Wiley Periodicals, Inc.


title: Questionnaire-based analysis of infection prevention and control in healthcare facilities in Saudi Arabia in regards to Middle East Respiratory Syndrome

text: Abstract Effective implementation of infection prevention and control in healthcare facilities depends on training, awareness and compliance of healthcare workers. In Saudi Arabia recent significant hospital outbreaks, including Middle East Respiratory Syndrome Coronavirus (MERS-CoV), have resulted from lack of, or breakdown in, infection prevention and control procedures. This study was designed to assess attitudes to, and awareness of, infection prevention and control policies and guidelines among healthcare workers of different professions and institution types in Saudi Arabia. A questionnaire was administered to 607 healthcare workers including physicians (n=133), nurses (n=162), laboratory staff (n=233) and other staff (n=79) in government hospitals, private hospitals and poly clinics. Results were compared using Chi square analysis according to profession type, institution type, age group and nationality (Saudi or non-Saudi) to assess variability. Responses suggested that there are relatively high levels of uncertainty among healthcare workers across a range of infection prevention and control issues, including institution-specific issues, surveillance and reporting standards, and readiness and competence to implement policies and respond to outbreaks. There was evidence to suggest that staff in private hospitals and nurses were more confident than other staff types. Carelessness of healthcare workers was the top-cited factor contributing to causes of outbreaks (65.07% of total group), and hospital infrastructure and design was the top-cited factor contributing to spread of infection in the hospital (54.20%), followed closely by lack and shortage of staff (53.71%) and no infection control training program (51.73%). An electronic surveillance system was considered the most effective by staff (81.22%). We have identified areas of concern among healthcare workers in Saudi Arabia on infection prevention and control which vary between institutions and among different professions. This merits urgent multi-factorial actions to try to ensure outbreaks such as MERS-CoV can be minimized and contained.


title: Automated Detection of Conformational Epitopes Using Phage Display Peptide Sequences

text: BACKGROUND: Precise determination of conformational epitopes of neutralizing antibodies represents a key step in the rational design of novel vaccines. A powerful experimental method to gain insights on the physical chemical nature of conformational epitopes is the selection of linear peptides that bind with high affinities to a monoclonal antibody of interest by phage display technology. However, the structural characterization of conformational epitopes from these mimotopes is not straightforward, and in the past the interpretation of peptide sequences from phage display experiments focused on linear sequence analysis to find a consensus sequence or common sequence motifs. RESULTS: We present a fully automated search method, EpiSearch that predicts the possible location of conformational epitopes on the surface of an antigen. The algorithm uses peptide sequences from phage display experiments as input, and ranks all surface exposed patches according to the frequency distribution of similar residues in the peptides and in the patch. We have tested the performance of the EpiSearch algorithm for six experimental data sets of phage display experiments, the human epidermal growth factor receptor-2 (HER-2/neu), the antibody mAb Bo2C11 targeting the C(2) domain of FVIII, antibodies mAb 17b and mAb b12 of the HIV envelope protein gp120, mAb 13b5 targeting HIV-1 capsid protein and 80R of the SARS coronavirus spike protein. In all these examples the conformational epitopes as determined by the X-ray crystal structures of the antibody-antigen complexes, were found within the highest scoring patches of EpiSearch, covering in most cases more than 50% residues of experimental observed conformational epitopes. Input options of the program include mapping of a single peptide or a set of peptides on the antigen structure, and the results of the calculation can be visualized on our interactive web server. AVAILABILITY: Users can access the EpiSearch from our web server http://curie.utmb.edu/episearch.html


title: Temporal dynamics, diversity, and interplay in three components of the virodiversity of a Mallard population: Influenza A virus, avian paramyxovirus and avian coronavirus

text: Abstract Multiple infections, or simultaneous infection of a host with multiple parasites, are the rule rather than the exception. Interactions between co-occurring pathogens in a population may be mutualistic, competitive or facilitative. For some pathogen combinations, these interrelated effects will have epidemiological consequences; however this is as yet poorly incorporated into practical disease ecology. For example, screening of Mallards for influenza A viruses (IAV) have repeatedly revealed high prevalence and large subtype diversity in the Northern Hemisphere. Other studies have identified avian paramyxovirus type 1 (APMV-1) and coronaviruses (CoVs) in Mallards, but without making inferences on the larger viral assemblage. In this study we followed 144 wild Mallards across an autumn season in a natural stopover site and constructed infection histories of IAV, APMV-1 and CoV. There was a high prevalence of IAV, comprising of 27 subtype combinations, while APMV-1 had a comparatively low prevalence (with a peak of 2%) and limited strain variation, similar to previous findings. Avian CoVs were common, with prevalence up to 12%, and sequence analysis identified different putative genetic lineages. An investigation of the dynamics of co-infections revealed a synergistic effect between CoV and IAV, whereby CoV prevalence was higher given that the birds were co-infected with IAV. There were no interactive effects between IAV and APMV-1. Disease dynamics are the result of an interplay between parasites, host immune responses, and resources; and is imperative that we begin to include all factors to better understand infectious disease risk.


title: Impact of an Infectious Disease Specialist on an Antimicrobial Stewardship Program at a Resource-Limited, Non-Academic Community Hospital in Korea

text: Background: Implementing a successful antimicrobial stewardship program (ASP) is difficult for non-academic community (NAC) hospitals due to insufficient infrastructure. Aim: We evaluated the impact of an infectious disease specialist (IDS) on implementing an ASP in a resource-limited setting in Korea. Methods: A retrospective study was performed at a NAC hospital between June 2015 and August 2018. An IDS has led an ASP at the hospital since June 2017. We used an interrupted time series analysis to evaluate longitudinal effects of the IDS-led ASP on the amount of antibiotic use and incidence of multidrug-resistant organism (MDRO) acquisition. Findings: Total antibiotic use changed from 698.82 ± 74.41 to 602.09 ± 69.94 defined daily dose/1000 patient-days (PDs) after intervention. An immediate reduction in the use of carbapenems, glycopeptides, penicillins, and other antibiotics followed the IDS-led ASP. The 3rd/4th generation cephalosporins and carbapenems prescription rates decreased in slope after the intervention. Incidence of MDRO acquisition changed from 1.38, 0.78, and 0.21/1000 PDs to 1.06, 0.15, and 0.32/1000 PDs in methicillin-resistant Staphylococcus aureus, multidrug-resistant Acinetobacter baumannii, and multidrug-resistant Pseudomonas aeruginosa, respectively. The incidence of methicillin-resistant Staphylococcus aureus and multidrug-resistant Acinetobacter baumannii acquisition immediately decreased following intervention. Conclusion: An IDS can implement a successful ASP by reducing antibiotic consumption and MDRO acquisition at resource-limited NAC hospitals.


title: The Differential Clinical Impact of Human Coronavirus Species in Children With Cystic Fibrosis

text: We investigated the clinical impact of human coronaviruses (HCoV) OC43, 229E, HKU1 and NL63 in pediatric patients with cystic fibrosis (CF) during routine and exacerbation visits. A total of 408 nasopharyngeal aspirate samples were obtained from 103 patients over a 1-year period. Samples positive for HCoV were submitted for nucleotide sequencing to determine the species. Nineteen samples (4.65%) were positive for HCoV, of which 8 were positive for NL63, 6 for OC43, 4 for HKU1, and 1 for 229E. Identification of HCoV was not associated with an increased rate of respiratory exacerbations, but NL63-positive patients had higher exacerbation rates than patients who were positive for other HCoV species.


title: Knowledge, risk perceptions, and preventive precautions among Hong Kong students during the 2009 influenza A (H1N1) pandemic

text: This study reports the knowledge, risk perceptions, and preventive precautions among 288 Hong Kong secondary school students during the 2009 influenza A (H1N1) pandemic. Only 28.1% of the students expressed knowledge of all 3 major routes of H1N1 transmission, and 58.1% perceived a high risk of H1N1 infection. Adoption of 4-5 preventive precautions was not prevalent (40.9%) and was significantly associated with female sex, better knowledge of transmission, and higher perceived risk of infection.


title: High-Mannose But Not Complex-Type Glycosylation of Tetherin Is Required for Restriction of HIV-1 Release

text: Tetherin is an interferon-inducible antiviral protein that inhibits the release of a broad spectrum of enveloped viruses by retaining virions at the surface of infected cells. While the role of specific tetherin domains in antiviral activity is clearly established, the role of glycosylation in tetherin function is not clear. In this study, we carried out a detailed investigation of this question by using tetherin variants in which one or both sites of N-linked glycosylation were mutated (N65A, N92A, and N65,92A), and chemical inhibitors that prevent glycosylation at specific stages of oligosaccharide were added or modified. The single N-linked glycosylation mutants, N65A and N92A, efficiently inhibited the release of Vpu-defective human immunodeficiency virus type 1 (HIV-1). In contrast, the non-glycosylated double mutant, N65,92A, lost its ability to block HIV-1 release. The inability of the N65,92A mutant to inhibit HIV-1 release is associated with a lack of cell-surface expression. A role for glycosylation in cell-surface tetherin expression is supported by tunicamycin treatment, which inhibits the first step of N-linked glycosylation and impairs both cell-surface expression and antiviral activity. Inhibition of complex-type glycosylation with kifunensine, an inhibitor of the oligosaccharide processing enzyme mannosidase 1, had no effect on either the cell-surface expression or antiviral activity of tetherin. These results demonstrate that high-mannose modification of a single asparagine residue is necessary and sufficient, while complex-type glycosylation is dispensable, for cell-surface tetherin expression and antiviral activity.


title: Prophylactic and therapeutic intranasal administration with an immunomodulator, Hiltonol® (Poly IC:LC), in a lethal SARS-CoV-infected BALB/c mouse model

text: Abstract Hiltonol®, (Poly IC:LC), a potent immunomodulator, is a synthetic, double-stranded polyriboinosinic-polyribocytidylic acid (poly IC) stabilized with Poly-L-lysine and carboxymethyl cellulose (LC). Hiltonol® was tested for efficacy in a lethal SARS-CoV-infected BALB/c mouse model. Hiltonol® at 5, 1, 0.5 or 0.25 mg/kg/day by intranasal (i.n.) route resulted in significant survival benefit when administered at selected times 24 h prior to challenge with a lethal dose of mouse-adapted severe acute respiratory syndrome coronavirus (SARS-CoV). The infected BALB/c mice receiving the Hiltonol® treatments were also significantly effective in protecting mice against weight loss due to infection (p < 0.001). Groups of 20 mice were dosed with Hiltonol® at 2.5 or 0.75 mg/kg by intranasal instillation 7, 14, and 21 days before virus exposure and a second dose was given 24 h later, prophylactic Hiltonol® treatments (2.5 mg/kg/day) were completely protective in preventing death, and in causing significant reduction in lung hemorrhage scores, lung weights and lung virus titers. Hiltonol® was also effective as a therapeutic when give up to 8 h post virus exposure; 100% of the-infected mice were protected against death when Hiltonol® was administered at 5 mg/kg/day 8 h after infection. Our data suggest that Hiltonol® treatment of SARS-CoV infection in mice leads to substantial prophylactic and therapeutic effects and could be used for treatment of other virus disease such as those caused by MERS-CoV a related coronavirus. These properties might be therapeutically advantageous if Hiltonol® is considered for possible clinical use.


title: A comparative analysis of some policy options to reduce rationing in the UK's NHS: Lessons from a general equilibrium model incorporating positive health effects

text: Abstract This paper seeks to determine the macro-economic impacts of changes in health care provision. The resource allocation issues have been explored in theory, by applying the Rybczynski theorem, and empirically, using a computable general equilibrium (CGE) model for the UK with a detailed health component. From the theory, changes in non-health outputs are shown to depend on factor-bias and scale effects, the net effects generally being indeterminate. From the applied model, a rise in the National Health Service (NHS) budget is shown to yield overall welfare gains, which fall by two-thirds assuming health care-specific factors. A nominally equivalent migration policy yields even higher welfare gains.


title: SARS-Cov-2RNA Found on Particulate Matter of Bergamo in Northern Italy: First Evidence

text: BACKGROUND: The burden of COVID-19 was extremely severe in Northern Italy, an area characterized by high concentrations of particulate matter (PM), which is known to negatively affect human health. Consistently with evidence already available for other viruses, we initially hypothesized the possibility of SARS-CoV-2 presence on PM, and we performed a first experiment specifically aimed at confirming or excluding this research hyphotesys. METHODS: We have colelcted 34 PM10 samples in Bergamo area (the epicenter of the Italian COVID-19 epidemic) by using two air samplers over a continuous 3-weeks period. Filters were properly stored and underwent RNA extraction and amplification according to WHO protocols in two parallel blind analyses performed by two different authorized laboratories. Up to three highly specific molecular marker genes (E, N, and RdRP) were used to test the presence of SARS-CoV-2 RNA on particulate matter. RESULTS: The first test showed positive results for gene E in 15 out of 16 samples, simultaneously displaying positivity also for RdRP gene in 4 samples. The second blind test got 5 additional positive results for at least one ofthe three marker genes. Overall, we tested 34 RNA extractions for the E, N and RdRP genes, reporting 20 positive results for at least one of the three marker genes, with positivity separately confirmed for all the three markers. Control tests to exclude false positivities were successfully accomplished. CONCLUSION: This is the first evidence that SARS-CoV-2 RNA can be present on PM, thus suggesting a possible use as indicator of epidemic recurrence.


title: A surface plasmon resonance-based assay for small molecule inhibitors of human cyclophilin A

text: Abstract A simple protocol for generating a highly stable and active surface plasmon resonance (SPR) sensor surface of recombinant human hexahistidine cyclophilin A (His-CypA) is described. The sensor surface was sensitive and stable enough to allow, for the first time, the screening and ranking of several novel small-molecule (M r ∼250–500Da) ligands in a competition binding assay with cyclosporin A (CsA). It also allowed us to accurately determine the kinetic rate constants for the interaction between His-CypA and CsA. His-CypA was first captured on a Ni2+–nitrilotriacetic acid (NTA) sensor chip and was then briefly covalently stabilized, coupling via primary amines. The significant baseline drift observed due to dissociation of weakly bound His-CypA from the Ni2+–NTA moiety was eliminated, resulting in a surface that was stable for at least 36h. In addition, immobilized protein activity levels were high, typically between 85 and 95%, assayed by the interaction between His-CypA and CsA. The mean equilibrium dissociation constant for CsA (K dCsA) binding to the immobilized His-CypA was 23±6nM, with on and off rates of 0.53±0.1μM−1 s−1 and 1.2±0.1 (×10−2) s−1, respectively. These values agree well with the values for the corresponding binding constants determined from steady-state and kinetic fluorescence titrations in solution.


title: New Digital Technologies for the Surveillance of Infectious Diseases at Mass Gathering Events

text: Outbreaks of infectious diseases at mass gatherings can strain the health system of the host region and pose a threat to local and global health. In addition to strengthening existing surveillance systems, most host nations also utilize novel technologies to assess disease risk and augment traditional surveillance approaches. We review novel approaches to disease surveillance utilizing the Internet, mobile phone applications, and wireless sensor networks. These novel approaches to disease surveillance can result in prompt detection.


Letter to the Editor in response to the article “COVID-19 and diabetes: Can DPP4 inhibition play a role?”


title: Cochrane Review: Interventions for the interruption or reduction of the spread of respiratory viruses

text: BACKGROUND: Viral epidemics or pandemics such as of influenza or severe acute respiratory syndrome (SARS) pose a significant threat. Antiviral drugs and vaccination may not be adequate to prevent catastrophe in such an event. OBJECTIVES: To systematically review the evidence of effectiveness of interventions to interrupt or reduce the spread of respiratory viruses (excluding vaccines and antiviral drugs, which have been previously reviewed). SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2006, issue 4); MEDLINE (1966 to November 2006); OLDMEDLINE (1950 to 1965); EMBASE (1990 to November 2006); and CINAHL (1982 to November 2006). SELECTION CRITERIA: We scanned 2300 titles, excluded 2162 and retrieved the full papers of 138 trials, including 49 papers of 51 studies. The quality of three randomised controlled trials (RCTs) was poor; as were most cluster RCTs. The observational studies were of mixed quality. We were only able to meta‐analyse case‐control data. We searched for any interventions to prevent viral transmission of respiratory viruses (isolation, quarantine, social distancing, barriers, personal protection and hygiene). Study design included RCTs, cohort studies, case‐control studies, cross‐over studies, before‐after, and time series studies. DATA COLLECTION AND ANALYSIS: We scanned the titles, abstracts and full text articles using a standardised form to assess eligibility. RCTs were assessed according to randomisation method, allocation generation, concealment, blinding, and follow up. Non‐RCTs were assessed for the presence of potential confounders and classified as low, medium, and high risk of bias. MAIN RESULTS: The highest quality cluster RCTs suggest respiratory virus spread can be prevented by hygienic measures around younger children. Additional benefit from reduced transmission from children to other household members is broadly supported in results of other study designs, where the potential for confounding is greater. The six case‐control studies suggested that implementing barriers to transmission, isolation, and hygienic measures are effective at containing respiratory virus epidemics. We found limited evidence that the more uncomfortable and expensive N95 masks were superior to simple surgical masks. The incremental effect of adding virucidals or antiseptics to normal handwashing to decrease respiratory disease remains uncertain. The lack of proper evaluation of global measures such as screening at entry ports and social distancing prevent firm conclusions about these measures. AUTHORS' CONCLUSIONS: Many simple and probably low‐cost interventions would be useful for reducing the transmission of epidemic respiratory viruses. Routine long‐term implementation of some of the measures assessed might be difficult without the threat of a looming epidemic. PLAIN LANGUAGE SUMMARY: Interventions to interrupt or reduce the spread of respiratory viruses Although respiratory viruses usually only cause minor disease, they can cause epidemics. Approximately 10% to 15% of people worldwide contract influenza annually, with attack rates as high as 50% during major epidemics. Global pandemic viral infections have been devastating because of their wide spread. In 2003 the severe acute respiratory syndrome (SARS) epidemic affected ˜8,000 people, killed 780, and caused an enormous social and economic crisis. A new avian influenza pandemic caused by the H5N1 strain might be more catastrophic. Single measures (particularly the use of vaccines or antiviral drugs) may be insufficient to interrupt the spread. We found 51 studies including randomised controlled trials (RCTs) and observational studies with a mixed risk of bias. Respiratory virus spread might be prevented by hygienic measures around younger children. These might also reduce transmission from children to other household members. Implementing barriers to transmission, isolation, and hygienic measures may be effective at containing respiratory virus epidemics. There was limited evidence that (more uncomfortable and expensive) N95 masks were superior to simple ones. Adding virucidals or antiseptics to normal handwashing is of uncertain benefit. There is insufficient evaluation of global measures such as screening at entry ports and social distancing.


docid: wwwqqvca

title: Pharmacoinformatics and molecular dynamics simulation studies reveal potential covalent and FDA-approved inhibitors of SARS-CoV-2 main protease 3CL(pro)

text: The SARS-CoV-2 was confirmed to cause the global pandemic of coronavirus disease 2019 (COVID-19). The 3-chymotrypsin-like protease (3CLpro), an essential enzyme for viral replication, is a valid target to combat SARS-CoV and MERS-CoV. In this work, we present a structure-based study to identify potential covalent inhibitors containing a variety of chemical warheads. The targeted Asinex Focused Covalent (AFCL) library was screened based on different reaction types and potential covalent inhibitors were identified. In addition, we screened FDA-approved protease inhibitors to find candidates to be repurposed against SARS-CoV-2 3CLpro. A number of compounds with significant covalent docking scores were identified. These compounds were able to establish a covalent bond (C–S) with the reactive thiol group of Cys145 and to form favorable interactions with residues lining the substrate-binding site. Moreover, paritaprevir and simeprevir from FDA-approved protease inhibitors were identified as potential inhibitors of SARS-CoV-2 3CLpro. The mechanism and dynamic stability of binding between the identified compounds and SARS-CoV-2 3CLpro were characterized by molecular dynamics (MD) simulations. The identified compounds are potential inhibitors worthy of further development as COVID-19 drugs. Importantly, the identified FDA-approved anti-hepatitis-C virus (HCV) drugs paritaprevir and simeprevir could be ready for clinical trials to treat infected patients and help curb COVID-19. Communicated by Ramaswamy H. Sarma


title: Viral inhibitors reveal overlapping themes in regulation of cell death and innate immunity

text: Abstract Many viruses regulate a crucial point in the apoptotic pathway by expressing viral Bcl-2 homologues, which have become useful tools to investigate the mechanisms behind the control of the mitochondrial checkpoint of apoptosis. Concurrently, a number of viral inhibitors of innate immune signalling have been instrumental tools in the discovery of key host pathways. Here we discuss how viral inhibitors of the apoptotic and innate signalling pathways have further enhanced the understanding of both research fields and are beginning to shed light on how these two pathways converge.


A call for new 4R’s based Radiation Oncology model in COVID-19 Pandemic


The Importance of a Centralized Line Service during the COVID-19 Pandemic


title: Impairment of the DeISGylation Activity of Foot-and-Mouth Disease Virus Lpro Causes Attenuation In Vitro and In Vivo

text: Foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) affects several pathways of the host innate immune response. Previous studies in bovine cells demonstrated that deletions (leaderless [LLV]) or point mutations in Lpro result in increased expression of interferon (IFN) and IFN-stimulated genes (ISGs), including, among others, the ubiquitin-like protein modifier ISG15 and the ubiquitin specific peptidase USP18. In addition to its conventional papain-like protease activity, Lpro acts as a deubiquitinase (DUB) and deISGylase. In this study, we identified a conserved residue in Lpro that is involved in its interaction with ISG15. Mutation W105A rendered Escherichia coli-expressed Lpro unable to cleave the synthetic substrate pro-ISG15 while preserving cellular eIF4G cleavage. Interestingly, mutant FMDV W105A was viable. Overexpression of ISG15 and the ISGylation machinery in porcine cells resulted in moderate inhibition of FMDV replication, along with a decrease of the overall state of ISGylation in wild-type (WT)-infected cells. In contrast, reduced deISGylation was observed upon infection with W105A and leaderless virus. Reduction in the levels of deubiquitination was also observed in cells infected with the FMDV LproW105A mutant. Surprisingly, similarly to WT, infection with W105A inhibited IFN/ISG expression despite displaying an attenuated phenotype in vivo in mice. Altogether, our studies indicate that abolishing/reducing the deISGylase/DUB activity of Lpro causes viral attenuation independently of its ability to block the expression of IFN and ISG mRNA. Furthermore, our studies highlight the potential of ISG15 to be developed as a novel biotherapeutic molecule against FMD. IMPORTANCE In this study, we identified an aromatic hydrophobic residue in foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) (W105) that is involved in the interaction with ISG15. Mutation in Lpro W105 (A12-LproW105A) resulted in reduced deISGylation in vitro and in porcine-infected cells. Impaired deISGylase activity correlated with viral attenuation in vitro and in vivo and did not affect the ability of Lpro to block expression of type I interferon (IFN) and other IFN-stimulated genes. Moreover, overexpression of ISG15 resulted in the reduction of FMDV viral titers. Thus, our study highlights the potential use of Lpro mutants with modified deISGylase activity for development of live attenuated vaccine candidates, and ISG15 as a novel biotherapeutic against FMD.


title: ChurchInAction: the role of religious interventions in times of COVID-19

text: In a recently published letter to the editor of this journal, the authors have called for the need to establish psychological support structures that cater to people’s mental health in this time of the coronavirus disease 2019 pandemic. To be more holistic, we extend this call to include people’s spiritual well-being as well. We highlight the initiatives of the Philippines’ religious sector. In particular, we report some of the interventions made by the Roman Catholic Church that have led to the social media hashtag, #ChurchInAction. These religious and spiritual interventions showcase the efforts of the Philippine Church and play an important role in providing assistance in time of public health crisis.


title: Molecular cloning and characterization of GuHMGR, an HMG-CoA reductase gene from liquorice (Glycyrrhiza uralensis)

text: A full length cDNA encoding HMGR (designated as GuHMGR) was isolated from liquorice (Glycyrrhiza uralensis) based on degenerated PCR and genome walking. The full length cDNA of GuHMGR was 2330 bp with a 1518-bp open reading frame (ORF) encoding a 505-aa polypeptide. Bioinformatics analysis indicated that there were two trans-membrane domains in GuHMGR. A molecular model of tertiary structure showed that GuHMGR is a novel HMGR with a similar spatial structure to other plant HMGRs. The deduced polypeptide of GuHMGR has an isoelectric point (pI) of 6.41 and a calculated molecular weight of about 54.7 kDa. Sequence comparison and phylogenetic tree analysis showed that GuHMGR had the highest homology with HMGRs from Pisum sativum and Medicago truncatula, indicating that GuHMGR belongs to the plant HMGR group. Expression analysis showed the similar amount of transcript level of GuHMGR in roots and leaves, suggesting that this gene was expressed constitutively in plants. Therefore, this novel HMGR gene would possibly provide a new strategy for studying the glycyrrhizin metabolism at the molecular level in the future.


Avoiding a new epidemic during a pandemic: the importance of assessing the risk of substance use disorders in the COVID-19 era.


title: Repurposing and reshaping of hospitals during the COVID-19 outbreak in South Korea

text: Abstract During the extensive outbreak of coronavirus disease 2019 (COVID-19) in South Korea, many strategies in the hospital setting, such as stratified patient care, the assignment of hospitals/beds by a task force team, and the establishment of dedicated COVID-19 hospitals, dedicated COVID-19 emergency centers, COVID-19 community facilities, and respiratory care split hospitals, were adopted to mitigate community transmission and prevent nosocomial infection. Most of these strategies were used during the Middle East Respiratory syndrome outbreak and were applied again successfully during the COVID-19 outbreak. The reallocation of health care capacity, repurposing of hospitals, and close collaboration between the government and the health care committee might have been the key to successfully addressing the crisis of COVID-19 given the shortage of health care resources.


title: First international external quality assessment of molecular diagnostics for Mers-CoV

text: Abstract Background Since the discovery of Middle East respiratory syndrome coronavirus (MERS-CoV) in 2012, diagnostic protocols were quickly published and deployed globally. Objectives We set out to assess the quality of MERS-CoV molecular diagnostics worldwide. Study design Both sensitivity and specificity were assessed using 12 samples containing different viral loads of MERS-CoV or common coronaviruses (OC43, 229E, NL63, HKU1). Results The panel was sent to more than 106 participants, of which 99 laboratories from 6 continents returned 189 panel results.Scores ranged from 100% (84 laboratories) to 33% (1 laboratory). 15% of respondents reported quantitative results, 61% semi-quantitative (Ct-values or time to positivity) and 24% reported qualitative results. The major specific technique used was real-time RT-PCR using the WHO recommended targets upE, ORF1a and ORF1b. The evaluation confirmed that RT-PCRs targeting the ORF1b are less sensitive, and therefore not advised for primary diagnostics. Conclusions The first external quality assessment MERS-CoV panel gives a good insight in molecular diagnostic techniques and their performances for sensitive and specific detection of MERS-CoV RNA globally. Overall, all laboratories were capable of detecting MERS-CoV with some differences in sensitivity. The observation that 8% of laboratories reported false MERS-CoV positive single assay results shows room for improvement, and the importance of using confirmatory targets.


docid: rzzohwof

title: Characterisation of different forms of the accessory gp3 canine coronavirus type I protein identified in cats

text: Abstract ORF3 is a supplemental open reading frame coding for an accessory glycoprotein gp3 of unknown function, only present in genotype I canine strain (CCoV-I) and some atypical feline FCoV strains. In these latter hosts, the ORF3 gene systematically displays one or two identical deletions leading to the synthesis of truncated proteins gp3-Δ1 and gp3-Δ2. As deletions in CoV accessory proteins have already been involved in tissue or host switch, studies of these different gp3 proteins were conducted in canine and feline cell. All proteins oligomerise through covalent bonds, are N-glycosylated and are maintained in the ER in non-infected but also in CCoV-II infected cells, without any specific retention signal. However, deletions influence their level of expression. In canine cells, all proteins are expressed with similar level whereas in feline cells, the expression of gp3-Δ1 is higher than the two other forms of gp3. None of the gp3 proteins modulate the viral replication cycle of heterologous genotype II CCoV in canine cell line, leading to the conclusion that the gp3 proteins are probably advantageous only for CCoV-I and atypical FCoV strains.


Loneliness and Isolation in Long-term Care and the Covid-19 pandemic


title: Lipid Metabolism as a Source of Druggable Targets for Antiviral Discovery against Zika and Other Flaviviruses

text: The Zika virus (ZIKV) is a mosquito-borne flavivirus that can lead to birth defects (microcephaly), ocular lesions and neurological disorders (Guillain-Barré syndrome). There is no licensed vaccine or antiviral treatment against ZIKV infection. The effort to understand the complex interactions of ZIKV with cellular networks contributes to the identification of novel host-directed antiviral (HDA) candidates. Among the cellular pathways involved in infection, lipid metabolism gains attention. In ZIKV-infected cells lipid metabolism attributed to intracellular membrane remodeling, virion morphogenesis, autophagy modulation, innate immunity and inflammation. The key roles played by the cellular structures associated with lipid metabolism, such as peroxisomes and lipid droplets, are starting to be deciphered. Consequently, there is a wide variety of lipid-related antiviral strategies that are currently under consideration, which include an inhibition of sterol regulatory element-binding proteins (SREBP), the activation of adenosine-monophosphate activated kinase (AMPK), an inhibition of acetyl-Coenzyme A carboxylase (ACC), interference with sphingolipid metabolism, blockage of intracellular cholesterol trafficking, or a treatment with cholesterol derivatives. Remarkably, most of the HDAs identified in these studies are also effective against flaviviruses other than ZIKV (West Nile virus and dengue virus), supporting their broad-spectrum effect. Considering that lipid metabolism is one of the main cellular pathways suitable for pharmacological intervention, the idea of repositioning drugs targeting lipid metabolism as antiviral candidates is gaining force.