title: Identification of unusual RNA folding patterns encoded by bacteriophage T4 gene 60

text: Abstract A 50-nucleotide (nt) untranslated region (coding gap sequence) that interrupts the amino acid coding sequence in T4 gene 60, plus an additional 5 nt upstream and another 3 nt downstream from the gap sequence, shows unusual folding patterns according to RNA structure prediction. A predicted highly stable and significant hairpin structure in the 5' half of the gap sequence and a plausible tertiary structural element computed in the 3' part of the gap sequence seem significant by statistical tests on the wild-type (wt) sequence. This feature is absent in insertion, deletion and substitution variants of the gap sequence, in which template activities are markedly lower than that of the wt. The proposed feature is consistent with currently available data showing that the translational bypass of the coding gap is correlated with a stop codon involved in a stem-loop structure folded in the gap sequence. We suggest that the role of this segment in “ribosomal bypass” of a portion of the mRNA sequence is a property of its special folded structure.


title: Inflammation inhibitors were remarkably up‐regulated in plasma of severe acute respiratory syndrome patients at progressive phase

text: Severe acute respiratory syndrome (SARS) is a severe infectious disease that has affected many countries and regions since 2002. A novel member of the coronavirus, SARS‐CoV, has been identified as the causative agent. However, the pathogenesis of SARS is still elusive. In this study, we used 2‐D DIGE and MS to analyze the protein profiles of plasma from SARS patients, in the search for proteomic alterations associated with the disease progression, which could provide some clues to the pathogenesis. To enrich the low‐abundance proteins in human plasma, two highly abundant proteins, albumin and IgG, were first removed. By comparing the plasma proteins of SARS patients with those of a normal control group, several proteins with a significant alteration were found. The up‐regulated proteins were identified as alpha‐1 acid glycoprotein, haptoglobin, alpha‐1 anti‐chymotrypsin and fetuin. The down‐regulated proteins were apolipoprotein A‐I, transferrin and transthyretin. Most of the proteins showed significant changes (up‐ or down‐regulated) in the progressive phase of disease, and there was a trend back to normal level during the convalescent phase. Among these proteins, the alterations of fetuin and anti‐chymotrypsin were further confirmed by Western blotting. Since all the up‐regulated proteins identified above are well‐known inflammation inhibitors, these results strongly suggest that the body starts inflammation inhibition to sustain the inflammatory response balance in the progression of SARS.


title: Avian IgY antibodies and their recombinant equivalents in research, diagnostics and therapy

text: Abstract The generation and use of avian antibodies is of increasing interest in a wide variety of applications within the life sciences. Due to their phylogenetic distance, mechanisms of immune diversification and the way in which they deposit IgY immunoglobulin in the egg yolk, chickens provide a number of advantages compared to mammals as hosts for immunization. These advantages include: the one-step purification of antibodies from egg yolk in large amounts facilitates having a virtually continuous supply; the epitope spectrum of avian antibodies potentially grants access to novel specificities; the broad absence of cross-reactivity with mammalian epitopes avoids assay interference and improves the performance of immunological techniques. The polyclonal nature of IgY antibodies has limited their use since avian hybridoma techniques are not well established. Recombinant IgY, however, can be generated from mammalian monoclonal antibodies which makes it possible to further exploit the advantageous properties of the IgY scaffold. Moreover, cloning and selecting the immune repertoire from avian organisms is highly efficient, yielding antigen-specific antibody fragments. The recombinant approach is well suited to circumvent any limitations of polyclonal antibodies. This review presents comprehensive information on the generation, purification, modification and applications of polyclonal and monoclonal IgY antibodies.


title: The Implication of Oxidative Stress and AMPK-Nrf2 Antioxidative Signaling in Pneumonia Pathogenesis

text: It is widely recognized that chemical, physical, and biological factors can singly or synergistically evoke the excessive production of oxidative stress in pulmonary tissue that followed by pulmonary lesions and pneumonia. In addition, metabolic and endocrine disorder-induced diseases such as diabetes and obesity often expressed higher susceptibility to pulmonary infections, and presented severe symptoms which increasing the mortality rate. Therefore, the connection between the lesion of the lungs and the metabolic/endocrine disorders is an interesting and essential issue to be addressed. Studies have noticed a similar pathological feature in both infectious pneumonia and metabolic disease-intercurrent pulmonary lesions, that is, from the view of molecular pathology, the accumulation of excessive reactive oxygen species (ROS) in pulmonary tissue accompanying with activated pro-inflammatory signals. Meanwhile, Adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) and nuclear factor erythroid-2-related factor 2 (Nrf2) signaling plays important role in metabolic/endocrine homeostasis and infection response, and it's closely associated with the anti-oxidative capacity of the body. For this reason, this review will start from the summary upon the implication of ROS accumulation, and to discuss how AMPK-Nrf2 signaling contributes to maintaining the metabolic/endocrine homeostasis and attenuates the susceptibility of pulmonary infections.


Health measures for the reopening of schools, colleges, high schools and nurseries


title: Biologie moléculaire et microbiologie clinique en 2007 Les applications et leur avenir – Partie 2

text: Résumé La biologie moléculaire est omniprésente en biologie médicale et plus particulièrement en microbiologie. De nombreux articles démontrent son importance tant dans le domaine du diagnostic que du pronostic, de l'évaluation thérapeutique, de l'épidémiologie ou des risques biologiques naturels ou non. La quantité considérable d'articles sur ce sujet n'apporte pas toujours une réponse évidente sur le rôle de la biologie moléculaire dans un laboratoire de microbiologie qu'il soit hospitalier ou non. Cette revue constitue une synthèse des apports de cette discipline en microbiologie. À partir de cet état des lieux, certaines questions se posent, par exemple: la biologie moléculaire constitue-t-elle un réel apport en microbiologie? Dans quelles indications prescrire un examen de biologie moléculaire? Les réponses ne sont pas toujours simples. Elles sont évidentes dans certains cas (l'hépatite C par exemple) et le sont moins dans d'autres, la tuberculose par exemple. Dans la première partie de l'article, nous avons parlé des généralités appliquées à la microbiologie. Dans cette deuxième partie, nous abordons certaines applications, reflets de l'importance prise par la biologie moléculaire en microbiologie. Abstract Molecular biology is omnipresent in medical biology especially in microbiology. Many papers report studies on its importance in diagnosis, prognosis, therapeutic field, epidemiology, natural (or not) biological risks alike. The important sum of reports on this subject does not always bring an obvious answer to the part that molecular biology could play in a microbiology laboratory either public or private. This article summarizes the contributions of this specialized field in microbiology and brings some questions on its contribution. Is molecular biology a real progress in microbiology? What are the main indications of molecular biology today in this field? According to the infectious particle, the answers are not always simple. They can be obvious in some cases (for instance, hepatitis C) or less obvious in other cases (tuberculosis for instance). In the first part of this article, we reported general remarks in molecular biology related to microbiology. The second part report some important applications underlining the important extent of molecular biology in this field.


title: Engineering of a Lectibody Targeting High-Mannose-Type Glycans of the HIV Envelope

text: High-mannose-type glycans (HMGs) are aberrantly enriched on HIV envelope glycoproteins. However, there is currently no drug selectively targeting HIV-associated HMGs. Here, we describe a novel HMG-targeting “lectibody,” a recombinant Fc-fusion protein comprising human IgG1 Fc and a novel actinohivin lectin variant (Avaren) obtained by structure-guided modifications for improved overall surface charge properties (AvFc). AvFc was engineered and produced using a rapid and scalable plant-based transient overexpression system. The lectibody exhibited potent antiviral activity against HIV-1 groups M and O primary viruses, as well as HIV-2 and simian immunodeficiency virus (SIV) strains, without affecting normal human blood cells. Furthermore, the lectibody induced Fc-mediated cell killing activity against HIV-1-infected cells and selectively recognized SIVmac239-infected macaque mesenteric lymph node cells in vitro. AvFc showed an extended serum half-life in rats and rhesus macaques, while no discernible toxicity was observed upon repeated systemic dosing in mice. These results highlight AvFc’s potential as a biotherapeutic targeting HIV-associated HMGs of cell-free virions, as well as productively infected cells, providing a foundation for new anti-HIV strategies. Efficient and cost-effective bioproduction in greenhouse facilities may open unique possibilities for further development of AvFc.


Transforming graduate nursing education during an era of social distancing: Tools from the field()


No more “social distancing” but practice physical separation


title: Germ warfare: What's eating you?

text: Superfast tests for infections could save thousands of lives – so why aren't doctors allowed to use them?


title: Beyond the science of covid-19

text: Models of the new coronavirus's spread are imperfect, so factors other than the science play an important role too, says David Adam


title: Financial Risk for COVID-19-like Respiratory Hospitalizations in Consumer-Directed Health Plans

text: INTRODUCTION: : This study aims to quantify out-of-pocket spending associated with respiratory hospitalizations for conditions similar to those caused by coronavirus disease 2019 (COVID-19) and to compare out-of-pocket spending differences among those enrolled in consumer-directed health plans and in traditional, low-deductible plans. METHODS: : This study used de-identified administrative claims from the OptumLabs® Data Warehouse (January 1, 2016–August 31, 2019) to identify patients with a respiratory hospitalization. It compared unadjusted out-of-pocket spending among consumer-directed health plan enrollees to traditional plan enrollees using difference of mean significance tests and repeated the analysis separately by age category and by calendar year quarter. These data were collected on a rolling basis by OptumLabs and were analyzed in March 2020. RESULTS: : Commercially insured consumer-directed health plan enrollees had significantly higher out-of-pocket spending than traditional plan enrollees and these differences were largest among younger populations. The largest difference in out-of-pocket spending occurred during the first half of the year. CONCLUSIONS: : Consumer-directed health plan enrollees may experience differential financial burden from a hospitalization related to COVID-19. Although some insurers are waiving cost-sharing payments for COVID-19 treatment, self-insured employers remain exempt. Policy responses to date may be insufficient to reduce the financial burden on consumer-directed health plans enrollees with respiratory hospitalizations related to COVID-19.


title: Justifications for Non-Consensual Medical Intervention: From Infectious Disease Control to Criminal Rehabilitation

text: A central tenet of medical ethics holds that it is permissible to perform a medical intervention on a competent individual only if that individual has given informed consent to the intervention. Yet it occasionally seems morally permissible to carry out non-consensual medical interventions on competent individuals for the purpose of infectious disease control (IDC). We describe two different moral frameworks that have been invoked in support of non-consensual IDC interventions and identify five desiderata that might be used to guide assessments of the moral permissibility of such interventions on either kind of fundamental justification. We then consider what these desiderata imply for the justifiability of carrying out non-consensual medical interventions that are designed to facilitate rehabilitation amongst serious criminal offenders. We argue that these desiderata suggest that a plausible case can be made in favor of such interventions.


title: Induction of robust immunity response in mice by dual-expression-system-based recombinant baculovirus expressing the capsid protein of porcine circovirus type 2

text: BACKGROUND: Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease that causes progressive weight loss, dyspnea, tachypnea, anemia, jaundice, and diarrhea in piglets. Although baculovirus is an enveloped virus that infects insects in nature, it has emerged as a vaccine vector, and we used it to develop a novel candidate vaccine for a preventive or therapeutic strategy to control PCV2 infections. METHODS: Immunoblotting analysis of recombinant baculovirus and immunofluorescent staining of baculovirus-infected cells were followed using anti-ORF2 monoclonal antibodies. The BALB/c mice were immunized intramuscularly with this baculovirus. The titers of antibodies were mensurated with a Cap-protein-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The IFN-γ response in splenocytes harvested from immunized mice was measured by ELISA. Student's t-test was used to compare immune responses of different groups. RESULTS: In this study, we successfully constructed a dual-expression-system-based recombinant baculovirus BV-GD-ORF2, which can display the PCV2 capsid (Cap) protein and VSV-G protein on the viral envelope and also expressing Cap protein on transduced mammalian cells, thereby functioning as both a subunit and a DNA vaccine. After infection, the Cap protein was expressed and displayed on the viral surface, as demonstrated with an indirect fluorescence assay and immunoblotting. The vaccination of mice with recombinant baculovirus BV-GD-ORF2 successfully induced robust Cap-protein-specific humoral and cellular immune responses. CONCLUSIONS: Our findings collectively demonstrate that the recombinant baculovirus BV-GD-ORF2 is a potential vaccine against PCV2 infections.


title: Pathogenic Leptospira spp. in bats: Molecular investigation in Southern Brazil

text: Abstract The present study aimed to investigate the frequency of pathogenic Leptospira spp. in Brazilian bats and to determine possible risk factors associated to it. Ninety two bats of 12 species were evaluated. Whole genomic DNA from kidneys was extracted and real-time PCR specific to pathogenic Leptospira spp. was applied. Association between the frequency of specimens positive for Leptospira spp. and sex, age, bat species or family, season of collection, geographic localization and feeding habits was evaluated. The results showed that 39.13% of analyzed bats were found positive for Leptospira spp. Nine bat species had at least one positive result. There was no association among the evaluated variables and frequency of pathogenic Leptospira spp. Although the limitations due to lack of Leptospira spp. isolation, leptospiral carriage was demonstrated in bats of different species from southern Brazil, which reinforces the need for surveillance of infectious agents in wild animals.


On Channel Activity of Synthetic Peptides Derived from Severe and Acute Respiratory Syndrome Coronavirus (SARS-CoV) E Protein


title: On facing the SARS-cov-2 (COVID-19) with combination of nanomaterials and medicine: Possible strategies and first challenges

text: Global health is facing the most dangerous situation regarding the novel severe acute respiratory syndrome called coronavirus 2 (SARS-CoV-2), which is widely known as the abbreviated COVID-19 pandemic. This is due to the highly infectious nature of the disease and its possibility to cause pneumonia induced death in approximately 6.89% of infected individuals (data until 27 April 2020). The pathogen causing COVID-19 is called severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which is believed to be originated from the Wuhan Province in China. Unfortunately, an effective and approved vaccine for SARS-CoV-2 virus is still not available, making the situation more dangerous and currently available medical care futile. This unmet medical need thus requires significant and very urgent research attention to develop an effective vaccine to address the SARS-CoV-2 virus. In this review, the state-of-the-art drug design strategies against the virus are critically summarized including exploitations of novel drugs and potentials of repurposed drugs. The applications of nanochemistry and general nanotechnology was also discussed to give the status of nanodiagnostic systems for COVID-19.


Key words to be adopted for COVID-19 research: A return to simple, large, randomized trials


title: Risque d’introduction et voies d’importation par l’homme de maladies infectieuses exotiques : cas particulier de l’émergence de bactéries pathogènes multirésistantes aux antibiotiques, importées en France à l’occasion de voyages internationaux ou du rapatriement de patients hospitalisés à l’étranger

text: SUMMARY The spread of multidrug-resistant bacteria has become a major problem in France in recent years, owing to increasing antibiotic exposure, growing international exchanges, repatriation of hospitalized French patients, and treatment of French and foreign travelers in French hospitals. This article examines how different pathogens may become endemic in France.


title: New emerging dermatological symptoms in coronavirus pandemic

text: COVID‐19 (Coronavirus Disease 2019) is a fatal disease that could lead to a serious respiratory illness. It is caused by SARS‐CoV‐2 virus. In December 2019, the first of human cases were identified in China and the infection spread quickly around the world. The World Health Organisation (WHO) declared the disease, a global pandemic on 11 March 2020, even as COVID‐19 rapidly spread across the world. According to clinical reports, fever, headache, cough and myalgia are common clinical symptoms. Many symptoms also occur, such as sputum formation, haemoptysis and diarrhea.


title: Advances in viral respiratory infections: new experimental models

text: The wide array of models available for the study of respiratory viral infections is extremely valuable for the development of novel therapeutic and prophylactic options against these highly prevalent diseases. In addition, through these models we have gathered considerable insight into the cellular and molecular mechanisms involved in the pathogenesis of these infections and the inflammatory and immune responses they elicit in the host. This article reviews new promising models introduced recently in this field. Section Editor: Alastair Stewart – University of Melbourne, Australia There has long been interest in the potential relationships between viral respiratory illness and chronic respiratory disease. However, until relatively recently there have been few attempts to develop animal models that address chronic influences of acute and chronic infections. Professor Piedimonte and colleagues provide a commentary on the impact of molecular biology on studies of viral infection and address the important issues that arise in selecting appropriate strains and species to model the course of infection, inflammation and long term sequelae.


title: Venous Thrombosis Among Critically Ill Patients With Coronavirus Disease 2019 (COVID-19)

text: This case series reports a systematic assessment of deep vein thrombosis among patients in an intensive care unit in France with severe coronavirus disease 2019 (COVID-19).


title: Wearing a N95 mask increases rescuer's fatigue and decreases chest compression quality in simulated cardiopulmonary resuscitation

text: OBJECTIVES: N95 mask is essential for healthcare workers dealing with the coronavirus disease 2019 (COVID-19). However, N95 mask causes discomfort breathing with marked reduction in air exchange. This study was designed to investigate whether the use of N95 mask affects rescuer's fatigue and chest compression quality during cardiopulmonary resuscitation (CPR). METHODS: After a brief review of CPR, each participant performed a 2-minute continuous chest compression on a manikin wearing N95 (N95 group, n = 40) or surgical mask (SM group, n = 40). Compression rate and depth, the proportions of correct compression rate, depth, complete chest recoil and hand position were documented. Participants' fatigue was assessed using Borg score. RESULTS: Significantly lower mean chest compression rate and depth were both achieved in the N95 group than in the SM group (p < 0.05, respectively). In addition, the proportion of correct compression rate (61 ± 19 vs. 75 ± 195, p = 0.0067), depth (67 ± 16 vs. 90 ± 14, p < 0.0001) and complete recoil (91 ± 16 vs. 98 ± 5%, p = 0.0248) were significantly decreased in the N95 group as compared to the SM group. At the end of compression, the Borg score in the N95 group was significantly higher than that in the SM group (p = 0.027). CONCLUSION: Wearing a N95 mask increases rescuer's fatigue and decreases chest compression quality during CPR. Therefore, the exchange of rescuers during CPR should be more frequent than that recommended in current guidelines when N95 masks are applied.


title: Occupant-based energy upgrades selection for Canadian residential buildings based on field energy data and calibrated simulations

text: Occupant behavior in residential buildings has a direct impact on the effectiveness of energy-saving measures. In order to realize a buildings’ carbon mitigation targets, the impact of individual occupancy profiles needs to be integrated with building simulation models. This paper introduces a decision support framework as a potential solution to make energy performance upgrade choices based on different occupancy profiles. This framework has been demonstrated through a case study of two single-family detached homes in Canada, which were highly instrumented with sensors for monitoring energy input and output. The case studies represented two common occupancy profiles-(1) a family of four (consisting of 2 working adults and 2 teenagers); and (2) a retired couple. Firstly, calibrated energy models were developed by using one-year energy use data collected through an intrusive load monitoring technique. Secondly, energy upgrade combinations were considered for each profile and tested for additional investment, payback period and greenhouse gas (GHG) emissions. Lastly, the most suitable combination of energy upgrade for each profile was ranked using a multi-criteria decision-making method (e.g., TOPSIS). Results indicated that the retired couple used more energy than the family of four and required energy upgrades with usually higher payback periods to achieve the same level of GHG emission reduction. The results of this research are timely for energy policymaking and developing best management practices, which need to be implemented along with the deployment of more stringent building standards and codes.


title: Nucleolar-cytoplasmic shuttling of PRRSV nucleocapsid protein: a simple case of molecular mimicry or the complex regulation by nuclear import, nucleolar localization and nuclear export signal sequences

text: Abstract The order Nidovirales, which includes the arteriviruses and coronaviruses, incorporate a cytoplasmic replication scheme; however, the nucleocapsid (N) protein of several members of this group localizes to the nucleolus suggesting that viral proteins influence nuclear processes during replication. The relatively small, 123 amino acid, N protein of porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus, presents an ideal model system for investigating the properties and mechanism of N protein nucleolar localization. The PRRSV N protein is found in both cytoplasmic and nucleolar compartments during infection and after transfection of gene constructs that express N-enhanced green fluorescent protein (EGFP) fusion proteins. Experiments using oligopeptides, truncated polypeptides and amino acid-substituted proteins have identified several domains within PRRSV N protein that participate in nucleo-cytoplasmic shuttling, including a cryptic nuclear localization signal (NLS) called NLS-1, a functional NLS (NLS-2), a nucleolar localization sequence (NoLS), as well as a possible nuclear export signal (NES). The purpose of this paper is to review our current understanding of PRRSV N protein shuttling and propose a shuttling scheme regulated by RNA binding and post-translational modification.


Bats, Coronaviruses, and Deforestation: Toward the Emergence of Novel Infectious Diseases?


title: Vicarious traumatization in the general public, members, and non-members of medical teams aiding in COVID-19 control

text: Abstract Since December 2019, more than 79,000 people have been diagnosed with infection of the Corona Virus Disease 2019 (COVID-19). A large number of medical staff was sent to Wuhan city and Hubei province to aid COVID-19 control. Psychological stress, especially vicarious traumatization caused by the COVID-19 pandemic, should not be ignored. To address this concern, the study employed a total of 214 general public and 526 nurses (i.e., 234 front-line nurses and 292 non-front-line nurses) to evaluate vicarious traumatization scores via a mobile app-based questionnaire. Front-line nurses are engaged in the process of providing care for patients with COVID-19. The results showed that the vicarious traumatization scores for front-line nurses including scores for physiological and psychological responses, were significantly lower than those of non-front-line nurses (P < 0.001). Interestingly, the vicarious traumatization scores of the general public were significantly higher than those of the front-line nurses (P < 0.001); however, no statistical difference was observed compared to the scores of non-front-line nurses (P > 0.05). Therefore, increased attention should be paid to the psychological problems of the medical staff, especially non-front-line nurses, and general public under the situation of the spread and control of COVID-19. Early strategies that aim to prevent and treat vicarious traumatization in medical staff and general public are extremely necessary.


title: Adenovirus vectors for high-efficiency gene transfer into mammalian cells

text: Characteristics such as versatility, stability and high-level expression make adenovirus vectors invaluable tools for the expression of transgenes in mammalian cells for the development of recombinant viral vaccines and for delivery of therapeutic genes.


title: Implementation of the IHR Joint External Evaluation: Taiwan's Experiences

text: In February 2016, the World Health Organization developed the Joint External Evaluation (JEE) tool to independently assess country capacity to prevent, detect, and respond to public health threats as part of the International Health Regulations (IHR) (2005) monitoring and evaluation framework. In light of this, the Taiwan government actively engaged at least 19 government agencies or institutions and voluntarily implemented the JEE. An External Assessment Team consisting of 6 US subject matter experts conducted the external evaluation, including site visits, from June 21 to July 1, 2016. The results, published on October 18, 2016, are useful and will be translated into actions and change in the system. Based on Taiwan's experiences, early stakeholder engagement and an experts' pre-JEE pilot visit would contribute to a successful JEE process.


title: Modeling Arboviral Infection in Mice Lacking the Interferon Alpha/Beta Receptor

text: Arboviruses are arthropod-borne viruses that exhibit worldwide distribution and are a constant threat, not only for public health but also for wildlife, domestic animals, and even plants. To study disease pathogenesis and to develop efficient and safe therapies, the use of an appropriate animal model is a critical concern. Adult mice with gene knockouts of the interferon α/β (IFN-α/β) receptor (IFNAR(−/−)) have been described as a model of arbovirus infections. Studies with the natural hosts of these viruses are limited by financial and ethical issues, and in some cases, the need to have facilities with a biosafety level 3 with sufficient space to accommodate large animals. Moreover, the number of animals in the experiments must provide results with statistical significance. Recent advances in animal models in the last decade among other gaps in knowledge have contributed to the better understanding of arbovirus infections. A tremendous advantage of the IFNAR(−/−) mouse model is the availability of a wide variety of reagents that can be used to study many aspects of the immune response to the virus. Although extrapolation of findings in mice to natural hosts must be done with care due to differences in the biology between mouse and humans, experimental infections of IFNAR(−/−) mice with several studied arboviruses closely mimics hallmarks of these viruses in their natural host. Therefore, IFNAR(−/−) mice are a good model to facilitate studies on arbovirus transmission, pathogenesis, virulence, and the protective efficacy of new vaccines. In this review article, the most important arboviruses that have been studied using the IFNAR(−/−) mouse model will be reviewed.


title: Co-infection of broilers with Ornithobacterium rhinotracheale and H9N2 avian influenza virus

text: BACKGROUND: Since 2008, a progressive pneumonia has become prevalent in broilers and laying hens. This disease occurrs the first day after hatching and lasts more than 30 days, resulting in approximately 70% morbidity and 30% mortality in broilers. The objective of this study was to isolate and identify the pathogens that are responsible for the progressive pneumonia and establish an animal model for drug screening. RESULTS: 193 serum samples were collected from 8 intensive farms from 5 provinces in China and analysed in the current research. Our clinical survey showed that 65.2% to 100% of breeding broilers, breeding layers, broilers and laying hens were seropositive for ORT antibodies. From 8 intensive farms, six ORT isolates were identified by PCR and biochemical assays, and two H9N2 viruses were isolated. Newcastle Disease Virus (NDV) and Infectious BronchitisVirus (IBV) were excluded. Typical pneumonia and airsacculitis were observed both in broilers inoculated intraperitoneally with an ORT isolate alone and in those co-infected with ORT and H9N2 virus isolates. Specifically, the survival rate was 30%, 20%, 70%, 50% and 90% in birds inoculated with ORT+H9N2 virus, ORT followed by H9N2 virus, H9N2 virus followed by ORT, and ORT or H9N2 virus alone, respectively. CONCLUSIONS: The results of this study suggest that ORT infections of domestic poultry have been occurring frequently in China. ORT infection can induce higher economic losses and mortality if H9N2 AIV is also present. Although the isolation of ORT and H9N2 virus has been reported previously, there have been no reported co-infections of poultry with these two pathogens. This is the first report of co-infection of broilers with ORT and H9N2 virus, and this co-infection is probably associated with the outbreak of broiler airsacculitis in China, which has caused extensive economic losses.


title: Organization of Patient Management and Fungal Epidemiology in Cystic Fibrosis

text: The achievement of a better life for cystic fibrosis (CF) patients is mainly caused by a better management and infection control over the last three decades. Herein, we want to summarize the cornerstones for an effective management of CF patients and to give an overview of the knowledge about the fungal epidemiology in this clinical context in Europe. Data from a retrospective analysis encompassing 66,616 samples from 3235 CF patients followed-up in 9 CF centers from different European countries are shown.


title: Hemorrhagic stroke and anticoagulation in COVID-19

text: Abstract Background and Purpose : Patients with the Coronavirus Disease of 2019 (COVID-19) are at increased risk for thrombotic events and mortality. Various anticoagulation regimens are now being considered for these patients. Anticoagulation is known to increase the risk for adverse bleeding events, of which intracranial hemorrhage (ICH) is one of the most feared. We present a retrospective study of 33 patients positive for COVID-19 with neuroimaging-documented ICH and examine anticoagulation use in this population. Methods : Patients over the age of 18 with confirmed COVID-19 and radiographic evidence of ICH were included in this study. Evidence of hemorrhage was confirmed and categorized by a fellowship trained neuroradiologist. Electronic health records were analyzed for patient information including demographic data, medical history, hospital course, laboratory values, and medications. Results : We identified 33 COVID-19 positive patients with ICH, mean age 61.6 years (range 37 to 83 years), 21.2% of whom were female. Parenchymal hemorrhages with mass effect and herniation occurred in 5 (15.2%) patients, with a 100% mortality rate. Of the remaining 28 patients with ICH, 7 (25%) had punctate hemorrhages, 17 (60.7%) had small- moderate size hemorrhages, and 4 (14.3%) had a large single site of hemorrhage without evidence of herniation. Almost all patients received either therapeutic dose anticoagulation (in 22 [66.7%] patients) or prophylactic dose (in 3 [9.1] patients) prior to ICH discovery. Conclusions : Anticoagulation therapy may be considered in patients with COVID-19 though the risk of ICH should be taken into account when developing a treatment regimen.


title: Molecular characterization of canine parvovirus and canine enteric coronavirus in diarrheic dogs on the island of St. Kitts: First report from the Caribbean region

text: Abstract Although canine parvovirus (CPV) and canine enteric coronavirus (CCoV) are important enteric pathogens of dogs and have been studied extensively in different parts of the world, there are no reports on these viruses from the Caribbean region. During 2015–2016, a total of 104 diarrheic fecal samples were collected from puppies and adult dogs, with or without hemorrhagic gastroenteritis, on the Caribbean island of St. Kitts (KNA). By PCR, 25 (24%, n=104) samples tested positive for CPV. Based on analysis of the complete deduced VP2 amino acid sequences, 20 of the KNA CPV strains were assigned to new CPV-2a (also designated as CPV-2a-297A). On the other hand, the VP2 genes of the remaining 5 strains were partially characterized, or could not be sequenced. New CPV-2a was the predominant CPV variant in St. Kitts, contrasting the molecular epidemiology of CPV variants reported in most studies from nearby North and South American countries. By RT-PCR, CCoVs were detected in 5 samples (4.8%, n=104). Based on analysis of partial M-protein gene, the KNA CCoV strains were assigned to CCoV-I genotype, and were closely related to CCoV-I strains from Brazil. To our knowledge, this is the first report on detection and genetic diversity of CPV and CCoV in dogs from the Caribbean region, and underscores the importance of similar studies in the other Caribbean islands.


title: A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

text: Abstract Canine parvovirus (CPV) is an important pathogen in domestic dogs, and the original antigenic types CPV-2 and its variants, CPV-2a, 2b and 2c, are prevalent worldwide. A multiplex TaqMan real-time PCR method was developed for the detection and differentiation of four antigenic types of CPV. A set of primers and probes, CPV-305F/CPV-305R and CPV-2-305P (for CPV-2)/CPV-2a-305P (for CPV-2a, 2b and 2c), was able to differentiate CPV-2 and its variants (CPV-2a, 2b and 2c). Another set of primers and probes, CPV-426F/CPV-426R and CPV-2-426P (for CPV-2 and 2a)/CPV-2b-426P (for CPV-2b)/CPV-2c-426P (for CPV-2c), was able to differentiate CPV-2a (2), CPV-2b, and CPV-2c. With these primers and probes, the multiplex TaqMan real-time PCR assay detected effectively and differentiated CPV-2, 2a, 2b and 2c by two separate real-time PCRs. No cross reactivity was observed with canine distemper virus, canine adenovirus, and canine coronavirus. The detection limit of the assay is 101 genome copies/μL for CPV-2, CPV-2a, CPV-2b, and 102 copies/μL for CPV-2c. The multiplex real-time PCR has 100% agreement with DNA sequencing. We provide a sensitive assay that simultaneously detects and differentiate four antigenic types of CPV and the method was also used for quantification of CPVs viral genome.


title: Quantitative RT Real Time PCR and indirect immunofluorescence for the detection of human parainfluenza virus 1, 2, 3

text: Abstract Human parainfluenza viruses (HPIVs) are distributed worldwide and are involved mainly in the pathogenesis of respiratory tract infections. The development and optimization of three quantitative reverse transcription real time polymerase chain reactions (RT Real Time Qt-PCRs) and an indirect immunofluorescence (IFA) for the detection and quantitation of HPIV-1, -2 and -3 in clinical samples are described. Efficiency, sensitivity, specificity, inter- and intra-assay variability and turnaround time of the two methods were compared. These assays have been validated on 131 bronchoalveolar lavage specimens. Based on the results obtained, the molecular methods represent a valid and rapid tool for clinical management and should be included in diagnostic panels aimed to evaluate suspected respiratory tract infections.


title: Severe Acute Respiratory Syndrome (SARS)

text: Abstract Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus (SARS-CoV), spread largely via droplets. Patients present with rapidly progressive pneumonia and respiratory failure. Diagnosis requires the demonstration of epidemiological linkage, clinical profiles, and virological evidence of SARS-CoV infection. Treatment of SARS is largely anecdotal. High-dose corticosteroid therapy might be useful for critically ill patients, but previous use of ribavirin (a broad-spectrum antiviral agent) was probably not efficacious. Some survivors of SARS suffer from avascular necrosis of hip and knee joints. Poor prognostic factors include advanced age, comorbidities, and high serum lactate dehydrogenase and neutrophil levels.


title: Induction of caspase-dependent apoptosis by betanodaviruses GGNNV and demonstration of protein α as an apoptosis inducer

text: Abstract Betanodaviruses, members of the Nodaviridae family, are the causative agents of viral nervous necrosis in fish and infection by which cause high mortality in larvae and juveniles in a wide range of marine fish species in Asia, Europe, Australia, Martinique, and Tahit. Greasy grouper (Epinephelus tauvina) nervous necrosis viruses (GGNNV) were investigated for their apoptotic activity in culture cells. GGNNV infection of sea bass (SB) cells appeared to induce a typical cytopathic effect (CPE), i.e., cytoplasmic vacuolation, thinning, rounding up, detachment of infected cells from the cultured dish, and eventually cell lysis and death. The infected SB cells underwent DNA fragmentation and stained positive in terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) assay, suggesting that GGNNV infection induced apoptosis in SB cells. In addition, GGNNV-infected SB cells showed an increased activity of caspase-8-like proteases (IETDase) and caspase-3-like proteases (IETDase), whereas inhibitor of caspase-8 and caspase-3 reduced GGNNV-induced apoptosis. This suggests that GGNNV may promote apoptosis via the extrinsic pathway in SB cells. Protein α, the precursor of GGNNV capsid proteins, was transiently expressed in SB and Cos-7 cells. The DNA fragmentation and TUNEL positive signal were apparent in SB and Cos-7 cells expressing protein α, suggesting that protein α may serve as an apoptotic inducer in these cells. Moreover, expression of protein α resulted in the activation of caspase-3-like proteases in both cells, which could be inhibited by a caspase-3-like protease specific inhibitor DEVD-CHO peptide. These results suggest that fish caspases are important elements in GGNNV-meditated apoptosis.


title: Chemical-informatics approach to COVID-19 drug discovery: Monte Carlo based QSAR, virtual screening and molecular docking study of some in-house molecules as papain-like protease (PLpro) inhibitors

text: World Health Organization characterized novel coronavirus disease (COVID-19), caused by severe acute respiratory syndrome (SARS) coronavirus-2 (SARS-CoV-2) as world pandemic. This infection has been spreading alarmingly by causing huge social and economic disruption. In order to response quickly, the inhibitors already designed against different targets of previous human coronavirus infections will be a great starting point for anti-SARS-CoV-2 inhibitors. In this study, our approach integrates different ligand based drug design strategies of some in-house chemicals. The study design was composed of some major aspects: (a) classification QSAR based data mining of diverse SARS-CoV papain-like protease (PLpro) inhibitors, (b) QSAR based virtual screening (VS) to identify in-house molecules that could be effective against putative target SARS-CoV PLpro and (c) finally validation of hits through receptor-ligand interaction analysis. This approach could be used to aid in the process of COVID-19 drug discovery. It will introduce key concepts, set the stage for QSAR based screening of active molecules against putative SARS-CoV-2 PLpro enzyme. Moreover, the QSAR models reported here would be of further use to screen large database. This study will assume that the reader is approaching the field of QSAR and molecular docking based drug discovery against SARS-CoV-2 PLpro with little prior knowledge. Communicated by Ramaswamy H. Sarma


title: The UPR sensor IRE1α and the adenovirus E3-19K glycoprotein sustain persistent and lytic infections

text: Persistent viruses cause chronic disease, and threaten the lives of immunosuppressed individuals. Here, we elucidate a mechanism supporting the persistence of human adenovirus (AdV), a virus that can kill immunosuppressed patients. Cell biological analyses, genetics and chemical interference demonstrate that one of five AdV membrane proteins, the E3-19K glycoprotein specifically triggers the unfolded protein response (UPR) sensor IRE1α in the endoplasmic reticulum (ER), but not other UPR sensors, such as protein kinase R-like ER kinase (PERK) and activating transcription factor 6 (ATF6). The E3-19K lumenal domain activates the IRE1α nuclease, which initiates mRNA splicing of X-box binding protein-1 (XBP1). XBP1s binds to the viral E1A-enhancer/promoter sequence, and boosts E1A transcription, E3-19K levels and lytic infection. Inhibition of IRE1α nuclease interrupts the five components feedforward loop, E1A, E3-19K, IRE1α, XBP1s, E1A enhancer/promoter. This loop sustains persistent infection in the presence of the immune activator interferon, and lytic infection in the absence of interferon.


title: Superinfection and cell regeneration can lead to chronic viral coinfections

text: Abstract Molecular diagnostic techniques have revealed that approximately 43% of the patients hospitalized with influenza-like illness are infected by more than one viral pathogen, sometimes leading to long-lasting infections. It is not clear how the heterologous viruses interact within the respiratory tract of the infected host to lengthen the duration of what are usually short, self-limiting infections. We develop a mathematical model which allows for single cells to be infected simultaneously with two different respiratory viruses (superinfection) to investigate the possibility of chronic coinfections. We find that a model with superinfection and cell regeneration has a stable chronic coinfection fixed point, while superinfection without cell regeneration produces only acute infections. This analysis suggests that both superinfection and cell regeneration are required to sustain chronic coinfection via this mechanism since coinfection is maintained by superinfected cells that allow slow-growing infections a chance to infect cells and continue replicating. This model provides a possible mechanism for chronic coinfection independent of any viral interactions via the immune response.


title: Evaluation of a recombinant measles virus expressing hepatitis C virus envelope proteins by infection of human PBL-NOD/Scid/Jak3null mouse

text: Abstract In this study, we infected NOD/Scid/Jak3null mice engrafted human peripheral blood leukocytes (hu-PBL-NOJ) with measles virus Edmonston B strain (MV-Edm) expressing hepatitis C virus (HCV) envelope proteins (rMV-E1E2) to evaluate the immunogenicity as a vaccine candidate. Although human leukocytes could be isolated from the spleen of mock-infected mice during the 2-weeks experiment, the proportion of engrafted human leukocytes in mice infected with MV (103–105 pfu) or rMV-E1E2 (104 pfu) was decreased. Viral infection of the splenocytes was confirmed by the development of cytopathic effects (CPEs) in co-cultures of splenocytes and B95a cells and verified using RT-PCR. Finally, human antibodies against MV were more frequently observed than E2-specific antibodies in serum from mice infected with a low dose of virus (MV, 100–101 pfu, and rMV-E1E2, 101–102 pfu). These results showed the possibility of hu-PBL-NOJ mice for the evaluation of the immunogenicity of viral proteins.


title: The making of new regionalism in the cross-boundary metropolis of Hong Kong–Shenzhen, China

text: In the age of globalization, new regionalism has become a new trend of regional development. This study examines the making of new regionalism in South China, using the case of Hong Kong–Shenzhen region. By tracing the process of Hong Kong and Shenzhen integration during the past three decades, the research finds three stages in Hong Kong–Shenzhen regionalization after the late 1970s, namely, the emergence of informal regional society from below, the transitional regional society and formal regional society, with reference to Hettne’s concept of regionness. Hong Kong’s internal economic, political and societal difficulties and external changing conditions are conducive to Hong Kong government’s re-regulation toward a more positive stance on cross-boundary regionalization. Although intense economic and social interactions exist between Hong Kong and Shenzhen, a cross-boundary regional community has not emerged yet. The slow development of cross-boundary regional community is different from European case. The active participation of the governments aims to facilitate cross-boundary economic and infrastructure development and planning. This can take place well before reaching higher level of regionness: the formation of a regional community or the regional institutionalized polity.


title: Barriers to Hospital Electronic Public Health Reporting and Implications for the COVID-19 Pandemic

text: We sought to identify barriers to hospital reporting of electronic surveillance data to local, state, and federal public health agencies and the impact on areas projected to be overwhelmed by the COVID-19 pandemic. Using 2018 American Hospital Association data, we identified barriers to surveillance data reporting, and combined this with data on the projected impact of the COVID-19 pandemic on hospital capacity at the hospital referral region (HRR) level. Our results find the most common barrier was public health agencies lacked the capacity to electronically receive data, with 41.2% of all hospitals reporting it. We also identified 31 HRRs in the top quartile of projected bed capacity needed for COVID-19 patients in which over half of hospitals in the area reported that the relevant public health agency was unable to receive electronic data. Public health agencies’ inability to receive electronic data is the most prominent hospital-reported barrier to effective syndromic surveillance. This reflects the policy commitment of investing in information technology for hospitals without a concomitant investment in IT infrastructure for state and local public health agencies.


title: Is the prone position indicated in critically ill patients with SARS-CoV-2 during the peri-operative period?

text: Abstract Coronaviruses including SARS-CoV-2 are a large family of viruses that cause illnesses ranging from the common cold to more severe diseases. A SARS-CoV-2 is a new strain that has not been previously identified in humans. The majority of critically ill patients admitted to intensive care units with confirmed severe infection with SARS-CoV-2 developed an acute respiratory distress like syndrome. The main objective of this opinion paper is to raise the discussion about the possible benefit of keeping the patient with COVID-19 disease and acute hypoxemic respiratory failure (AHRF) in the prone position during the perioperative period, especially where this position is not a required factor for the surgical or invasive procedure. We believe that the prone position, due to its favorable pulmonary physiology, can improve the V̇/Q̇ ratio in the perioperative period.


title: Zika Vaccine Development: Current Status

text: Abstract Zika virus outbreaks have been explosive and unpredictable and have led to significant adverse health effects—as well as considerable public anxiety. Significant scientific work has resulted in multiple candidate vaccines that are now undergoing further clinical development, with several vaccines now in phase 2 clinical trials. In this review, we survey current vaccine efforts, preclinical and clinical results, and ethical and other concerns that directly bear on vaccine development. It is clear that the world needs safe and effective vaccines to protect against Zika virus infection. Whether such vaccines can be developed through to licensure and public availability absent significant financial investment by countries, and other barriers discussed within this article, remains uncertain.


title: Infection by rhinovirus: Similarity of clinical signs included in the case definition of influenza IAn/H1N1

text: INTRODUCTION: Although new influenza virus (IAn/H1N1) infections are mild and indistinguishable from any other seasonal influenza virus infections, there are few data on comparisons of the clinical features of infection with (IAn/H1N1) and with other respiratory viruses. The incidence, clinical aspects and temporal distribution of those respiratory viruses circulating during flu pandemic period were studied. METHODS: Respiratory samples from patients with acute influenza-like symptoms were collected from May 2009 to December 2009. Respiratory viruses were detected by conventional culture methods and genome amplification techniques. RESULTS: Although IAn/H1N1 was the virus most frequently detected, several other respiratory viruses co-circulated with IAn/H1N1 during the pandemic period, especially rhinovirus. The similarity between clinical signs included in the clinical case definition for influenza and those caused by other respiratory viruses, particularly rhinovirus, suggest that a high percentage of viral infections were clinically diagnosed as case of influenza. CONCLUSIONS: Our study offers useful information to face future pandemics caused by influenza virus, indicating that differential diagnoses are required in order to not overestimate the importance of the pandemic.


title: Association between nitric oxide synthesis and vaccination-acquired resistance to murine hepatitis virus by spf mice

text: Abstract Murine hepatitis virus strain 3 (MHV-3) produces a strain-dependent pattern of disease, with A/J and BALB/c mice being considered models of resistance and susceptibility, respectively. A role for nitric oxide in controlling infection remains debatable; thus, we monitored nitric oxide levels in blood and liver of immunized and nonimmunized spf mice during infection by electron paramagnetic resonance. In parallel, liver histology, virus titers, and plasma alanine aminotransferase (ALT) activity were monitored. Nitric oxide synthesis was barely detectable in BALB/c mice, which showed a progressive increase in virus titers and ALT activity. These animals died with a shorter survival time than A/J mice. The latter displayed a less severe infection and presented detectable levels of nitric oxide as nitrosyl complexes in blood and liver at 72 hpi. Immunized mice from both strains became resistant to MHV-3 and showed comparable levels of nitrosyl complexes in blood and liver at an early time (24 hpi). Thereafter, nitric oxide levels decreased but remained detectable in blood up to 96 hpi. Immunized mice were capable of clearing the virus and clearance was inhibited by administration of a nitric oxide synthase inhibitor. Overall, the results support a role for nitric oxide in controlling MHV-3 infection.


title: Molecular epidemiology of bovine toroviruses circulating in South Korea

text: Abstract The prevalence of the bovine torovirus (BToV) and its genetic characterization have been reported in North America, Europe and Japan. Therefore, this study examined the prevalence and genetic diversity of the BToV in a total of 645 diarrheic fecal samples from 629 Korean native beef calf herds using RT-PCR and nested PCR with the primer pairs specific to a part of the BToV membrane (M) gene. Overall, 19 (2.9%) out of 645 diarrheic samples from 19 herds (6.9%) tested positive for BToVs by either RT-PCR or nested PCR. A comparison of the nucleotide (nt) and amino acid (aa) sequences of a part of the BToV M gene (409bp) among the BToVs showed the Korean BToVs to have comparatively higher sequence homology to the Japanese and Dutch BToVs than to the American and Italian BToVs. Generally, the Korean BToV strains clustered with the Japanese and Dutch BToV strains. However, the American and Italian BToV strains clustered on a separate major branch, suggesting that these are more distantly related to other known BToV strains. These results suggest that the BToV infections are sporadic in diarrheic calves in South Korea, and the Korean BToV strains are more closely related to the Japanese and Dutch BToVs than to the American and Italian BToVs.


title: Chapter 7 Bronchus-Associated Lymphoid Tissue (BALT) Structure and Function

text: Abstract Bronchus-associated lymphoid tissue (BALT) is a constitutive mucosal lymphoid tissue adjacent to major airways in some mammalian species, including rats and rabbits, but not humans or mice. A related tissue, inducible BALT (iBALT), is an ectopic lymphoid tissue that is formed upon inflammation or infection in both mice and humans and can be found throughout the lung. Both BALT and iBALT acquire antigens from the airways and initiate local immune responses and maintain memory cells in the lungs. Here, we discuss the development and function of BALT and iBALT in the context of pulmonary immunity to infectious agents, tumors, and allergens as well as autoimmunity and inflammatory diseases of the lung.


title: Distributions of time, place, and population of novel coronavirus disease 2019 (COVID-19) from January 20 to February 10, 2020, in China

text: Abstract Background and objective Since December 2019, increasing cases of novel coronavirus disease 2019 (COVID-19) are being detected worldwide. The purpose of this paper is to provide a scientific reference for the global prevention and control of COVID-19. Methods General demographic characteristics, epidemiological history, and clinical symptoms of COVID-19 were collected that had been reported on the websites of multiple Municipal Health Commissions in China. We herein describe distributions in time, place, and population of COVID-19. Results As of midnight on February 10, 2020, the number of confirmed cases of COVID-19 in China was 42,638, and the province with the largest number of confirmed cases was Hubei (31728), followed by Guangdong (1177), Zhejiang (1117), and Henan (1105) province. The number of cases and the speed of confirmed cases in provinces other than Hubei were more moderate than those of the Hubei province. The median (interquartile range) age of 1740 patients with COVID-19 was 44 (33, 54) years, with a range of 10 months to 89 years. Conclusions The COVID-19 epidemic should be considered a global threat and the steps for control include early diagnosis and treatment, as well as isolation.


title: A Critical Care and Transplantation-Based Approach to Acute Respiratory Failure after Hematopoietic Stem Cell Transplantation in Children

text: Abstract Acute respiratory failure contributes significantly to nonrelapse mortality after allogeneic hematopoietic stem cell transplantation. Although there is a trend of improved survival over time, mortality remains unacceptably high. An understanding of the pathophysiology of early respiratory failure, opportunities for targeted therapy, assessment of the patient at risk, optimal use of noninvasive positive pressure ventilation, strategies to improve alveolar recruitment, appropriate fluid management, care of the patient with chronic lung disease, and importantly, a team approach between critical care and transplantation services may improve outcomes.


title: Indian agriculture, air pollution, and public health in the age of COVID

text: Emerging evidence supports the intuitive link between chronic health conditions associated with air pollution and the vulnerability of individuals and communities to COVID-19. Poor air quality already imposes a highly significant public health burden in Northwest India, with pollution levels spiking to hazardous levels in November and early December when rice crop residues are burned. The urgency of curtailing the COVID-19 pandemic and mitigating a potential resurgence later in the year provides even more justification for accelerating efforts to dramatically reduce open agricultural burning in India.


title: Age-dependent gender differences of COVID-19 in mainland China: comparative study

text: BACKGROUND: The ongoing pandemic of novel coronavirus disease 2019 (COVID-19) is challenging global public health system. Sex-differences in infectious diseases are a common but neglected problem. METHODS: We used the national surveillance database of COVID-19 in mainland China to compared gender differences in attack rate (AR), proportion of severe and critical cases (PSCC) and case fatality rate (CFR) in relation to age, affected province, and onset-to-diagnosis interval. RESULTS: The overall AR was significantly higher in female population than in males (63.9 versus 60.5 per million persons; P ˂ .001). By contrast, PSCC and CFR were significantly lower among female patients (16.9% and 4.0%) than among males (19.5% and 7.2%), with ORs of 0.87 and 0.57, respectively (both P ˂ .001). The female-to-male differences were age-dependent, which were significant among people aged 50–69 years for AR, and in the patients of 30-years or older for both PSCC and CFR (all P ≤ .001). The AR, PSCC and CFR varied greatly from province to province. However, female-to-male differences in AR, PSCC and CFR were significant in the epicenter, Hubei province, where 82.2% confirmed cases and 97.4% deaths occurred. After adjusting for age, affected province and onset-to-diagnosis interval, the female-to-male difference in AR, PSCC and CFR remained significant in multivariate logistic regression analyses. CONCLUSIONS: We elucidate an age-dependent gender dimorphism for COVID-19, in which the females have higher susceptibility but lower severity and fatality. Further epidemiological and biological investigations are required to better understand the sex-specific differences for effective interventions.


title: Role of Regulatory T Cells in Pathogenesis and Biological Therapy of Multiple Sclerosis

text: Multiple sclerosis (MS) is an inflammatory disease in which the myelin sheaths around the axons of the brain and spinal cord are damaged, leading to demyelination and scarring as well as a broad spectrum of signs and symptoms. It is caused by an autoimmune response to self-antigens in a genetically susceptible individual induced by unknown environmental factors. Principal cells of the immune system that drive the immunopathological processes are T cells, especially of T(H)1 and T(H)17 subsets. However, in recent years, it was disclosed that regulatory T cells took part in, too. Subsequently, there was endeavour to develop ways how to re-establish their physiological functions. In this review, we describe known mechanisms of action, efficacy, and side-effects of contemporary and emerging MS immunotherapeutical agents on Treg cells and other cells of the immune system involved in the immunopathogenesis of the disease. Furthermore, we discuss how laboratory immunology can offer physicians its help in the diagnosis process and decisions what kind of biological therapy should be used.


Challenges and Insights amidst the Covid-19 pandemic: Nutrition, the immune system and disease risk


Bringing Urban Legends into the Classroom


title: Evaluation of glycoprotein E subunit and live attenuated varicella‐zoster virus vaccines formulated with a single‐strand RNA‐based adjuvant

text: INTRODUCTION: Varicella‐zoster virus (VZV), a human alphaherpesvirus 3, elicits both chickenpox and shingles and/or postherpetic neuralgia. A live attenuated vaccine (LAV) and glycoprotein E (gE) subunit vaccine were developed to prevent VZV‐induced diseases. We recently reported that single‐strand RNA (ssRNA) based on the intergenic region of the internal ribosome entry site of cricket paralysis virus (CrPV) is an effective adjuvant for protein‐based and virus‐like particle‐based vaccines. Here, Chinese hamster ovary expression system and an LAV from Oka/SK strains. METHODS: We appraised the adjuvant effect of the same CrPV ssRNA encoding the gE gene formulated in the two vaccines using VZV‐primed C57BL/6 mice and guinea pigs. Humoral immunity and cell‐mediated immunity were assessed by enzyme‐linked immunosorbent assay (ELISA) and ELISPOT in gE subunit vaccine and by ELISA and fluorescent antibody to membrane antigen in LAV. RESULTS: The gE subunit vaccine‐induced gE‐specific antibodies and CD4(+) T‐cell responses (indicated by interferon‐γ [IFN‐γ] and interleukin‐2 secretion) in the ssRNA‐based adjuvant containing the VZV gE gene. Therefore, an ssRNA adjuvant combined with gE antigen can trigger the innate immune response and induce an adaptive immune response to ultimately activate humoral and cell‐mediated responses. VZV LAV could also induce VZV‐specific antibodies and IFN‐γ stimulated by LAV, whereas the effect of ssRNA as a vaccine adjuvant could not be confirmed. However, the ssRNA adjuvant increased VZV‐specific neutralizing antibody response. CONCLUSIONS: Taken together, these results highlight that the gE subunit vaccine and LAV developed in this study can be functional VZV vaccines, and ssRNAs appear to function better as adjuvants in a subunit vaccine than in an LAV.


title: Characteristics of community-acquired respiratory viruses infections except seasonal influenza in transplant recipients and non-transplant critically ill patients

text: Abstract Background/Purpose Transplant recipients are vulnerable to life-threatening community-acquired respiratory viruses (CA-RVs) infection (CA-RVI). Even if non-transplant critically ill patients in intensive care unit (ICU) have serious CA-RVI, comparison between these groups remains unclear. We aimed to evaluate clinical characteristics and mortality of CA-RVI except seasonal influenza A/B in transplant recipients and non-transplant critically ill patients in ICU. Methods We collected 37,777 CA-RVs multiplex real-time reverse transcription-polymerase chain reaction test results of individuals aged ≥18 years from November 2012 to November 2017. The CA-RVs tests included adenovirus, coronavirus 229E/NL63/OC43, human bocavirus, human metapneumovirus, parainfluenza virus 1/2/3, rhinovirus, and respiratory syncytial virus A/B. Results We found 286 CA-RVI cases, including 85 solid organ transplantation recipients (G1), 61 hematopoietic stem cell transplantation recipients (G2), and 140 non-transplant critically ill patients in ICU (G3), excluding those with repeated isolation within 30 days. Adenovirus positive rate and infection cases were most prominent in G2 (p < 0.001). The median time interval between transplantation and CA-RVI was 30 and 20 months in G1 and G2, respectively. All-cause in-hospital mortality was significantly higher in G3 than in G1 or G2 (51.4% vs. 28.2% or 39.3%, p = 0.002, respectively). The mechanical ventilation (MV) was the independent risk factor associated with all-cause in-hospital mortality in all three groups (hazard ratio, 3.37, 95% confidence interval, 2.04–5.56, p < 0.001). Conclusions This study highlights the importance of CA-RVs diagnosis in transplant recipients even in long-term posttransplant period, and in non-transplant critically ill patients in ICU with MV.


title: Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19

text: Abstract Recently, a novel coronavirus (SARS-CoV-2; coronavirus disease 2019, COVID-19) has emerged, rapidly spreading and severely straining the capacity of the global health community. Many nations are employing combinations of containment and mitigation strategies, where early diagnosis of COVID-19 is vital in controlling illness progression and limiting viral spread within the population. Thus, rapid and accurate methods of early detection are vital to contain COVID-19 and prevent further spread and predicted subsequent infectious waves of viral recurrence in future. Immediately after its initial characterization, Chinese and American Centers for Disease Control and Prevention (CDCs) rapidly employed molecular assays for detection of COVID-19, mostly employing real-time polymerase chain reaction (RT-PCR) methods. However, such methods require specific expensive items of equipment and highly trained analysts, requiring upwards of 4-8 hours to process. These requirements coupled with associated financial pressures may prevent effective deployment of such diagnostic tests. Loop mediated isothermal amplification(LAMP) is method of nucleic acid amplification which exhibits increased sensitivity and specificity are significantly rapid, and do not require expensive reagents or instruments, which aids in cost reduction for coronavirus detection. Studies have shown the successful application of LAMP assays in various forms to detect coronavirus RNA in patient samples, demonstrating that 1-10 copies of viral RNA template per reaction are sufficient for successful detection, ∼100-fold more sensitive than conventional RT-PCR methods. Importantly, studies have also now demonstrated the effectiveness of LAMP methodology in the detection of SARS-CoV-2 RNA at significantly low levels, particularly following numerous improvements to LAMP assay protocols. We hypothesise that recent advancements in enhanced LAMP protocols assay perhaps represent the best chance for a rapid and robust assay for field diagnosis of COVID-19, without the requirement of specialized equipment and highly trained professionals to interpret results. Herein, we present our arguments with a view to disseminate such findings, to assist the combat of this virus that is proving so devastating. We hope that this strategy could be applied rapidly, and confirmed for viability with clinical samples, before being rolled out for mass-diagnostic testing in these current times.


title: Nucleocapsid protein of SARS-CoV activates the expression of cyclooxygenase-2 by binding directly to regulatory elements for nuclear factor-kappa B and CCAAT/enhancer binding protein

text: Abstract SARS-associated coronavirus (SARS-CoV) causes inflammation and damage to the lungs resulting in severe acute respiratory syndrome. To evaluate the molecular mechanisms behind this event, we investigated the roles of SARS-CoV proteins in regulation of the proinflammatory factor, cyclooxygenase-2 (COX-2). Individual viral proteins were tested for their abilities to regulate COX-2 gene expression. Results showed that the COX-2 promoter was activated by the nucleocapsid (N) protein in a concentration-dependent manner. Western blot analysis indicated that N protein was sufficient to stimulate the production of COX-2 protein in mammalian cells. COX-2 promoter mutations suggested that activation of COX-2 transcription depended on two regulatory elements, a nuclear factor-kappa B (NF-κB) binding site, and a CCAAT/enhancer binding protein (C/EBP) binding site. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) demonstrated that SARS-CoV N protein bound directly to these regulatory sequences. Protein mutation analysis revealed that a Lys-rich motif of N protein acted as a nuclear localization signal and was essential for the activation of COX-2. In addition, a Leu-rich motif was found to be required for the N protein function. A sequence of 68 residuals was identified as a potential DNA-binding domain essential for activating COX-2 expression. We propose that SARS-CoV N protein causes inflammation of the lungs by activating COX-2 gene expression by binding directly to the promoter resulting in inflammation through multiple COX-2 signaling cascades.


title: Epidemic Surveillance of Covid-19: Considering Uncertainty and Under-Ascertainment

text: Epidemic surveillance is a fundamental part of public health practice. Addressing under-ascertainment of cases is relevant in most surveillance systems, especially in pandemics of new diseases with a large spectrum of clinical presentations as it may influence timings of policy implementation and public risk perception. From this perspective, this article presents and discusses early evidence on under-ascertainment of COVID-19 and its motifs, options for surveillance, and reflections around their importance to tailor public health measures. In the case of COVID-19, systematically addressing and estimating under-ascertainment of cases is essential to tailor timely public health measures, and communicating these findings is of the utmost importance for policy making and public perception.


title: Crystal Structures of the Main Peptidase from the SARS Coronavirus Inhibited by a Substrate-like Aza-peptide Epoxide

text: The main peptidase (Mpro) from the coronavirus (CoV) causing severe acute respiratory syndrome (SARS) is one of the most attractive molecular targets for the development of anti-SARS agents. We report the irreversible inhibition of SARS-CoV Mpro by an aza-peptide epoxide (APE; k inact/K i=1900(±400)M−1 s−1). The crystal structures of the Mpro:APE complex in the space groups C2 and P212121 revealed the formation of a covalent bond between the catalytic Cys145 Sγ atom of the peptidase and the epoxide C3 atom of the inhibitor, substantiating the mode of action of this class of cysteine-peptidase inhibitors. The aza-peptide component of APE binds in the substrate-binding regions of Mpro in a substrate-like manner, with excellent structural and chemical complementarity. In addition, the crystal structure of unbound Mpro in the space group C2 revealed that the “N-fingers” (N-terminal residues 1 to 7) of both protomers of Mpro are well defined and the substrate-binding regions of both protomers are in the catalytically competent conformation at the crystallization pH of 6.5, contrary to the previously determined crystal structures of unbound Mpro in the space group P21.


title: What Does the COVID-19 Pandemic Mean for Rheumatology Patients?

text: PURPOSE OF REVIEW: The COVID-19 outbreak has resulted in uncertainty for patients with autoimmune rheumatic diseases for several reasons. They are concerned about their risk of developing COVID-19 as many are immune suppressed from their disease and/or treatment, whether they should stop their advanced therapies, if they will have a worse outcome if/when infected due to their underlying medication condition(s) and if they will have drug availability, especially with press (without much data) coverage suggesting hydroxychloroquine may be used in COVID-19 infection causing diversion of medication supply. This article discusses how the pandemic affects people with systemic autoimmune rheumatic diseases. RECENT FINDINGS: Preliminarily, articles seem to suggest that patients with rheumatic diseases may not have more infections from SARS-CoV-2 and similar outcomes to age and gender matched patients, but fear of rheumatic medications increasing their risk, drug shortages, and work exposure all are concerns for patients. RECENT FINDINGS: The long term effects of the pandemic in patients with rheumatic diseases will not be known until much later and likely include stressors flaring disease (fear, illness, job loss, social isolation), post-traumatic stress, flaring due to stopping medications, less physician visits with subsequent under-treatment, and increases in chronic concomitant fatigue, pain, fibromyalgia.


title: Natural Marine Antiviral Products

text: Abstract The oceans are unique resources that provide a diverse array of natural products, primarily from invertebrates such as sponges, tunicates, bryozoans and molluscs and from marine bacteria and cyanobacteria. Pharmacologic research with marine chemicals continue to contribute potentially novel chemical leads in the ongoing global search for therapeutic agents in the treatment of multiple diverse categories. As infectious diseases evolve and develop resistance to existing pharmaceuticals, the marine environment provides novel leads against fungal, parasitic, bacterial and viral diseases. Limitations in our current antiviral treatment options and the continuing emergence of new pathogenic viruses have contributed to a growing need for new and effective chemotherapeutic agents to treat viral diseases. The search for potent antiviral agents is urgent in view of the dramatic situation of the global human immunodeficiency virus (HIV) epidemic, a possible spread of avian influenza and of other viral diseases. Effective antiviral therapeutics are not available, and the presently approved therapy for HIV has been recognized to be toxic, unable to eradicate the causative virus, and to induce severe drug resistance. The marine environment provides a rich source of chemical diversity for the screening and identification of new compounds with desirable antiviral properties. Antiviral testing has revealed numerous compounds from structural classes including polysaccharides, terpenoids, steroids, alkaloids and peptides that potentially inhibit both RNA and DNA viruses. This review presents an account of some research directed toward the discovery of new antiviral agents from marine sources.


title: The Accessory Protein ORF3 Contributes to Porcine Epidemic Diarrhea Virus Replication by Direct Binding to the Spike Protein

text: The porcine epidemic diarrhea virus (PEDV) is an important swine pathogen responsible for severe watery diarrhea, particularly in neonatal piglets. Despite extensive studies performed to elucidate the function of several viral proteins, the contribution of an accessory protein ORF3 in PEDV replication is still largely unknown. Here, we constructed expression plasmids as well as recombinant PEDV carrying myc-tagged ORF3 to assess their expression and subcellular localization in both transfected and infected cells. In PEDV-infected cells, ORF3 was predominantly localized in the cytoplasm, partially in the endoplasmic reticulum (ER) and the Golgi apparatus (Golgi). Interestingly, ORF3 with the N-terminal Flag tag was also detected on the cell surface concomitant with the spike (S) protein as determined by flow cytometry and confocal microscopy. ORF3 and S proteins were also co-localized at perinuclear compartments and in the vesicle-like structures in transfected and infected cells. We also demonstrated that both full-length and naturally truncated ORF3 proteins could interact with the S protein but with different binding affinity, which correlate with the ability of the protein to regulate virus replication in cell culture. Collectively, our results underscore the unprecedented role of the ORF3, which involves the interaction of ORF3 with S and, possibly, other structural protein during PEDV replication.


title: Highlights in Antiviral Drug Research: Antivirals at the Horizon

text: This review highlights ten “hot topics” in current antiviral research: (i) new nucleoside derivatives (i.e., PSI‐352938) showing high potential as a direct antiviral against hepatitis C virus (HCV); (ii) cyclopropavir, which should be further pursued for treatment of human cytomegalovirus (HCMV) infections; (iii) North‐methanocarbathymidine (N‐MCT), with a N‐locked conformation, showing promising activity against both α‐ and γ‐herpesviruses; (iv) CMX001, an orally bioavailable prodrug of cidofovir with broad‐spectrum activity against DNA viruses, including polyoma, adeno, herpes, and pox; (v) favipiravir, which is primarily pursued for the treatment of influenza virus infections, but also inhibits the replication of other RNA viruses, particularly (‐)RNA viruses such as arena, bunya, and hanta; (vi) newly emerging antiarenaviral compounds which should be more effective (and less toxic) than the ubiquitously used ribavirin; (vii) antipicornavirus agents in clinical development (pleconaril, BTA‐798, and V‐073); (viii) natural products receiving increased attention as potential antiviral drugs; (ix) antivirals such as U0126 targeted at specific cellular kinase pathways [i.e., mitogen extracellular kinase (MEK)], showing activity against influenza and other viruses; and (x) two structurally unrelated compounds (i.e., LJ‐001 and dUY11) with broad‐spectrum activity against virtually all enveloped RNA and DNA viruses. © 2012 Wiley Periodicals, Inc. Med. Res. Rev., 00, No. 0, 1–34, 2012


title: Daily emotional well‐being during the COVID‐19 pandemic

text: The COVID‐19 outbreak has become one of the largest public health crises of our time. Governments have responded by implementing self‐isolation and physical distancing measures that have profoundly impacted daily life throughout the world. In this study, we aimed to investigate how people experience the activities, interactions, and settings of their lives during the pandemic. The sample (N = 604) was assessed in Ireland on the 25 March 2020, following the closure of schools and non‐essential businesses. We examined within‐person variance in emotional well‐being and how people spend their time. We found that while most time was spent in the home (74%), time spent outdoors (8%) was associated with markedly raised positive affect and reduced negative emotions. Exercising, going for walks, gardening, pursuing hobbies, and taking care of children were the activities associated with the greatest affective benefits. Home‐schooling children and obtaining information about COVID‐19 were ranked lowest of all activities in terms of emotional experience. These findings highlight activities that may play a protective role in relation to well‐being during the pandemic, the importance of setting limits for exposure to COVID‐19‐related media coverage, and the need for greater educational supports to facilitate home‐schooling during this challenging period.


title: COVID-19 and Liver.

text: The current pandemic coronavirus labelled as Severe Acute Respiratory Distress Syndrome Coronavirus -2 (SARS -CoV-2) is a significant public health threat over for past few weeks. Overall case fatality rates range between 2-6%; however, the rates are higher in patients with severe disease, advanced age and underlying comorbidities like diabetes, hypertension and heart disease. Recent reports showed about 2-11% of patients with COVID-19 had underlying chronic liver disease. Experience from previous SARS epidemic suggest that 60% of patients developed various degrees of liver damage. In the current pandemic, hepatic dysfunction was seen in 14-53% of patients with COVID-19, particularly in those with severe disease. Cases of acute liver injury have been reported, associated with higher mortality. Hepatic involvement in COVID-19 could be multifactorial related to any of direct cytopathic effect of the virus, uncontrolled immune reaction, sepsis or drug induced liver injury. The postulated mechanism of viral entry is through the host ACE2 receptors that are abundantly present in type 2 alveolar cells. Interestingly, the expression of ACE2 receptors were identified in the gastrointestinal tract, vascular endothelium and cholangiocytes of the liver. Liver transplant recipients with COVID-19 have been reported recently. Effects of COVID-19 on underlying chronic liver disease requires a detailed evaluation and currently data is lacking and further research is warranted in this area. With lack of definitive therapy, patient education, hand hygiene and social distancing appears to be the cornerstone in minimising the disease spread.


title: Identification of co-infection by rotavirus and parvovirus in dogs with gastroenteritis in Mexico

text: This is the first report on circulating canine rotavirus in Mexico. Fifty samples from dogs with gastroenteritis were analyzed used polymerase chain reaction and reverse transcription polymerase chain reaction in order to identify parvovirus and rotavirus, respectively; 7% of dogs were infected with rotavirus exclusively, while 14% were co-infected with both rotavirus and parvovirus; clinical signs in co-infected dogs were more severe.


title: IMPACT OF CORONAVIRUS SYNDROMES ON PHYSICAL AND MENTAL HEALTH OF HEALTH CARE WORKERS: SYSTEMATIC REVIEW AND META-ANALYSIS

text: BACKGROUND: Health care workers (HCW) are at high risk of developing physical/mental health outcomes related to coronavirus syndromes. Nature and frequency of these outcomes are undetermined. METHODS: PRISMA/MOOSE-compliant (PROSPERO-CRD42020180205) systematic review of Web of Science/grey literature until 15th April 2020, to identify studies reporting physical/mental health outcomes in HCW infected/exposed to Severe Acute Respiratory Syndrome -SARS-, Middle East Respiratory Syndrome -MERS-, Novel coronavirus -COVID-19-. Proportion random effect meta-analyses, I(2) statistic, quality assessment and sensitivity analysis. RESULTS: 115 articles were included (n=60,458 HCW, age 36.1±7.1, 77.1% female). Physical health outcomes: 75.9% HCW infected by SARS/MERS/COVID-19 reported fever (95%CI=65.9%-83.7%, k=12, n=949), 47.9% cough (95%CI=39.2%-56.8%, k=14, n=970), 43.6% myalgias (95%CI=31.9%-56.0%, k=13, n=898), 42.3% chills (95%CI=20.2%-67.9%, k=7, n=716), 41.2% fatigue (95%CI=18.2%-68.8%, k=6, n=386), 34.6% headaches (95%CI=23.1%-48.2%, k=11, n=893), 31.2% dyspnoea (95%CI=23.2%-40.5%, k=12, n=1003), 25.3% sore throat (95%CI=18.8%-33.2%, k=8, n=747), 22.2% nausea/vomiting (95%CI=14.9%-31.8%, k=6, n=662), 18.8% diarrhoea (95%CI=11.9%-28.4%, k=9, n=824). Mental health outcomes: 62.5% HCW exposed to SARS/MERS/COVID-19 reported general health concerns (95%CI=57.0%-67,8%, k=2, n=2254), 43.7% fear (95%CI=33.9%-54.0%, k=4, n=584), 37.9% insomnia (95%CI=30.9%-45.5%, k=6, n=5067), 37.8% psychological distress (95%CI=28.4%-48.2%, k=15, n=24,346), 34.4% burnout (95%CI=19.3%-53.5%, k=3, n=1337), 29.0% anxiety features (95%CI=14.2%-50.3%, k=6, n=9191), 26.3% depressive symptoms (95%CI=12.5%-47.1%, k=8, n=9893), 20.7% post-traumatic stress disorder features (95%CI=13.2%-31%, k=11, n=3826), 16.1% somatisation (95%CI=0.2%-96.0%, k=2, n=2184), 14.0% stigmatisation feelings (95%CI=6.4%-28.1%, k=2, n=411). LIMITATIONS: Limited amount of evidence for some outcomes and suboptimal design in several studies included. CONCLUSIONS: SARS/MERS/COVID-19 have a substantial impact on the physical and mental health of HCW, which should become a priority for public health strategies.


title: Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein

text: The emergence of SARS-CoV-2 has resulted in >90,000 infections and >3,000 deaths. Coronavirus spike (S) glycoproteins promote entry into cells and are the main target of antibodies. We show that SARS-CoV-2 S uses ACE2 to enter cells and that the receptor-binding domains of SARS-CoV-2 S and SARS-CoV S bind with similar affinities to human ACE2, correlating with the efficient spread of SARS-CoV-2 among humans. We found that the SARS-CoV-2 S glycoprotein harbors a furin cleavage site at the boundary between the S(1)/S(2) subunits, which is processed during biogenesis and sets this virus apart from SARS-CoV and SARS-related CoVs. We determined cryo-EM structures of the SARS-CoV-2 S ectodomain trimer, providing a blueprint for the design of vaccines and inhibitors of viral entry. Finally, we demonstrate that SARS-CoV S murine polyclonal antibodies potently inhibited SARS-CoV-2 S mediated entry into cells, indicating that cross-neutralizing antibodies targeting conserved S epitopes can be elicited upon vaccination.


title: Airborne disinfection using microwave-based technology: Energy efficient and distinct inactivation mechanism compared with waterborne disinfection

text: Abstract Microwave has been extensively applied to inactivate microorganisms in liquids, food, and surfaces. However, energy efficiency is a limiting factor for the environmental application. The utilization pathway and energy efficiency of the microwave in different media have not been investigated. In this study, the inactivation performance, energy utilization, and bactericidal mechanisms for microwave-irradiated airborne and waterborne Escherichia coli were compared. A Beer-Lambert law-based model was also developed and validated to compare the inactivation performance in different phases. Microwave had greater inactivation effect on airborne bacteria than waterborne bacteria. The inactivation rate constant for airborne E. coli (0.29 s−1) was nearly 20 times higher than that of waterborne species (0.014 s−1). Most of the absorbed microwave energy (92.3%) was converted to increase water temperature instead of inactivating the waterborne bacteria, because the microwave photons were easily absorbed by water molecules. By contrast, 45.4% of the absorbed energy could disinfect the airborne bacteria. Finally, the required energies for 1-log inactivation were calculated as 2.3 J and 116.9 J per log-inactivation for airborne and waterborne E. coli, respectively. The airborne and waterborne E. coli samples showed distinct microwave inactivation mechanisms. Waterborne E. coli disinfection was primarily due to thermal effect, while the non-thermal effect was the major mechanism for airborne E. coli inactivation.


Venous thromboembolism and Covid-19


title: Anosmia: a COVID-19 symptom that must be investigated by rhinoplasty surgeons

text: the prevention of the COVID-19 epidemic between the LTCFs having been contaminated by at least one case of COVID-19 among residents and/or LTCFs staff and LTCFs with no cases of COVID between March 23rd and May 6th, in the year 2020 Methods Participants The epidemic risk being variable over time and from one region to another, and because the guidance have evolved over time in short-term periods, our study was limited to all the LTCFs of the Haute-Garonne department (Occitania region), SouthWestern France, between March 23rd and May 6th, 2020 We sent a questionnaire to LTCFs directors/medical staff by email on March 30th, to all the 132 LTCFs registered in the Haute-Garonne department After sending the questionnaire, we systematically performed a phone call (period of 6 to 19 May 2020) to the LTCFs either coordinating nurse or coordinating doctor in order to explain the questions and guide them on how to complete the questionnaire online The questionnaire focused on the preventive measures implemented in the LTCF before March 23rd, 2020 This date refers to the first confirmed case of COVID-19 in LTCFs in the department of Haute Garonne At that period, none of the other LTCFs had confirmed cases of COVID-19 The questionnaire therefore relates to practices at the time that preceded the arrival of the epidemic wave This date was also chosen because it was, on the basis of our field knowledge (7), an important motivational element for local LTCFs to implement more strictly all the guidance for LTCFs This event was indeed widely diffused in the media in the Haute Garonne department, in the official website of the association of LTCFs coordinating doctors and allowed a temporal reference of a major local event helping LTCFs staff to complete the questionnaire accurately Variables of interest The questionnaire included the status of the LTC (public, private for-profit, private non-profit) and the presence of a coordinating physician (yes/no) The implementation of the guidance were listed (each recommendation was a dichotomy: yes/no) regarding: the systematic wearing of masks by the healthcare professional;access to effective masks (surgical and/or FFP2);satisfactory supply of masks;satisfactory supply of hydro-alcoholic solute;access to specific internal or external training on hygiene measures throughout the year;the use of containment in residents' rooms;during meals, separating residents to each other, respecting a distance of at least one meter (in the dining room with distance, in small groups, in bedroom);cessation of group activities for residents (with distance, in small groups);the use of interim jobs;staff compartmentalization within zones (organization of the work so that the team works in small groups in one area of the LTCF with no physical connection with the other members of the team);resident compartmentalization within zones (organization of the LTCF so that the resident live in small groups in one area of the LTCF with no possible physical connection with the other residents);specific dressing procedure at the entrance (complete daily routine dress change at the entrance and exit of the LTCF for anyone entering the LTCF) In order to judge the global prevention effort made by the healthcare team, the questionnaire ended with a self-assessment scale: To what extent do you think the nursing home staff has satisfactorily applied all the recommended «barrier» measures? (rated from 0, not at all, to 10, perfectly) At the end of the phone call, the coordinating nurse or coordinating doctor was asked to give a global judgment of the follow-up of the guidance by the all nursing home staff Outcome measure The coordinating nurse or doctor should specify whether, in the LTCFs, one or more residents and/or one or more caregivers were infected with COVID-19 (yes/no) The cases had to be confirmed by a positive rt-PCR test Unity of measurement was the LTCF Ethics This study is a survey of the leaders of the LTCFs and no data relating to residents was collected The approval of the ethics c mmittee was not required Statistical Analysis Qualitative variables are described as numbers and percentage [n (%)] and quantitative variables as mean and standard deviation (SD) Bivariate analysis used the chi-square test (Fisher's exact test if applicable) and the Student's ttest for independent samples, as appropriate In order to consider the confounding factors, a logistic regression was realized Multicollinearity was assessed The initial model was composed of variables found associated with a LTCFs infected by COVID-19 to a threshold of 0 20 in bivariate analysis Then, a reduced model was performed, using a step by step backward regression Analysis has been duplicated separately for private and public LTCFs Results Among the 132 LTCFs in the region, 124 (93 9%) agreed to participate and 8 refused In 30 LTCFs, confirmed cases of COVID-19 (24 2%) were found among: one or more residents (n = 6 LTCFs), one or more caregivers (n = 17), or both (n = 7) Seven LTCFs (5 6%) had confirmed cases for two or more residents Table 1 shows the implementation of guidance in the LTCFs with confirmed COVID cases vs those without confirmed cases In bivariate analyses, LTCFs ownership, staff compartmentalization within zones, use of professional interim, and organization of the meals with ≥ 1-meter distance separating residents to each other were significantly associated with infection of COVID-19 in the LTCFs The


title: Blocking transmission of Middle East respiratory syndrome coronavirus (MERS-CoV) in llamas by vaccination with a recombinant spike protein

text: The ongoing Middle East respiratory syndrome coronavirus (MERS-CoV) outbreaks pose a worldwide public health threat. Blocking MERS-CoV zoonotic transmission from dromedary camels, the animal reservoir, could potentially reduce the number of primary human cases. Here we report MERS-CoV transmission from experimentally infected llamas to naïve animals. Directly inoculated llamas shed virus for at least 6 days and could infect all in-contact naïve animals 4–5 days after exposure. With the aim to block virus transmission, we examined the efficacy of a recombinant spike S1-protein vaccine. In contrast to naïve animals, in-contact vaccinated llamas did not shed infectious virus upon exposure to directly inoculated llamas, consistent with the induction of strong virus neutralizing antibody responses. Our data provide further evidence that vaccination of the reservoir host may impede MERS-CoV zoonotic transmission to humans.


title: Diminished production of type-I interferons and interleukin-2 in patients with multiple sclerosis

text: Abstract Several lines of evidence have supported the role of immunological mechanisms in the pathogenesis of multiple sclerosis (MS) and new immunomodulatory strategies for its treatment, e.g. subcutaneous application of interferon (IFN)-β, have emerged. We investigated the ability of peripheral blood mononuclear cells (PBMC) in 21 consecutive patients with clinically definite MS to produce interferons and lymphokines in response to viral or mitogenic stimulation. Ten patients showed clinical signs of disease activity (acute relapse) and 11 patients were in a stable condition. Additionally, white blood count, leukocyte differentiation and lymphocyte subtyping were performed. A group of age-related healthy blood donors served as control (n=20). There was no difference between patients and controls in the production of IFN-γ, tumor necrosis factor (TNF)-α and soluble interleukin (IL)-2 receptor. IFN-α and IFN-β responsiveness, however, was significantly lower in patients with stable disease than in patients with active disease and controls (p<0.001). Furthermore, secretion of IL-2 after stimulation was significantly diminished in both patient groups as compared to the control group (p<0.01). Analysis of T-cell subsets revealed a significantly lower amount of CD8+ T-cells in patients with stable disease, leading to a significantly higher CD4/CD8 ratio in this group as compared to patients with active disease. Our study depicted an IL-2 deficiency in MS patients which is shared with other autoimmune diseases. In addition, our findings suggest that the ability to produce type-I IFNs, IFN-α and IFN-β, is primarily impaired in MS patients and changes in correlation to the course of disease activity.


title: Kinetics and synergistic effects of siRNAs targeting structural and replicase genes of SARS-associated coronavirus

text: SARS-associated coronavirus was identified as the etiological agent of severe acute respiratory syndrome and a large virus pool was identified in wild animals. Virus generates drug resistance through fast mutagenesis and escapes antiviral treatment. siRNAs targeting different genes would be an alternative for overcoming drug resistance. Here, we report effective siRNAs targeting structural genes (i.e., spike, envelope, membrane, and nucleocapsid) and their antiviral kinetics. We also showed the synergistic effects of two siRNAs targeting different functional genes at a very low dose. Our findings may pave a way to develop cost effective siRNA agents for antiviral therapy in the future.


title: Identification and characterization of vp7 gene in Bombyx mori cytoplasmic polyhedrosis virus

text: Abstract The genome of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) contains 10 double stranded RNA segments (S1–S10). The segment 7 (S7) encodes 50kDa protein which is considered as a structural protein. The expression pattern and function of p50 in the virus life cycle are still unclear. In this study, the viral structural protein 7 (VP7) polyclonal antibody was prepared with immunized mouse to explore the presence of small VP7 gene-encoded proteins in Bombyx mori cytoplasmic polyhedrosis virus. The expression pattern of vp7 gene was investigated by its overexpression in BmN cells. In addition to VP7, supplementary band was identified with western blotting technique. The virion, BmCPV infected cells and midguts were also examined using western blotting technique. 4, 2 and 5 bands were detected in the corresponding samples, respectively. The replication of BmCPV genome in the cultured cells and midgut of silkworm was decreased by reducing the expression level of vp7 gene using RNA interference. In immunoprecipitation experiments, using a polyclonal antiserum directed against the VP7, one additional shorter band in BmCPV infected midguts was detected, and then the band was analyzed with mass spectrum (MS), the MS results showed thatone candidate interacted protein (VP7 voltage-dependent anion-selective channel-like isoform, VDAC) was identified from silkworm. We concluded that the novel viral product was generated with a leaky scanning mechanism and the VDAC may be an interacted protein with VP7.