title: Display technologies: application for the discovery of drug and gene delivery agents

text: Recognition of molecular diversity of cell surface proteomes in disease is essential for the development of targeted therapies. Progress in targeted therapeutics requires establishing effective approaches for high-throughput identification of agents specific for clinically relevant cell surface markers. Over the past decade, a number of platform strategies have been developed to screen polypeptide libraries for ligands targeting receptors selectively expressed in the context of various cell surface proteomes. Streamlined procedures for identification of ligand-receptor pairs that could serve as targets in disease diagnosis, profiling, imaging and therapy have relied on the display technologies, in which polypeptides with desired binding profiles can be serially selected, in a process called biopanning, based on their physical linkage with the encoding nucleic acid. These technologies include virus/phage display, cell display, ribosomal display, mRNA display and covalent DNA display (CDT), with phage display being by far the most utilized. The scope of this review is the recent advancements in the display technologies with a particular emphasis on molecular mapping of cell surface proteomes with peptide phage display. Prospective applications of targeted compounds derived from display libraries in the discovery of targeted drugs and gene therapy vectors are discussed.


title: The major histocompatibility complex influences the ethiopathogenesis of MS-like disease in primates at multiple levels

text: Abstract Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease primarily affecting the central nervous system. Of the many candidate polymorphic major histocompatibility complex (MHC) and non-MHC genes contributing to disease susceptibility, including those encoding effector (cytokines and chemokines) or receptor molecules within the immune system (MHC, TCR, Ig or FcR), human leukocyte antigen (HLA) class II genes have the most significant influence. In this article we put forward the hypothesis that the influence of HLA genes on the risk to develop MS is actually the sum of multiple antigen presenting cell (APC) and T-cell interactions involving HLA class I and class II molecules. This article will also discuss that, because of the genetic and immunologic similarity to humans, autoimmune models of MS in non-human primates are the experimental models “par excellence” to test this hypothesis.


title: SARS‐CoV‐2 was found in the bile juice from a patient with severe COVID‐19

text: SARS‐CoV‐2 has infected over 170,000 people worldwide and was associated with substantial mortality. Previous studies have reported that the SARS‐CoV‐2 could be detected in sputum, faeces, urine and other specimens from COVID‐19 patients. In the case report, the novel coronavirus was found in the bile specimen from a patient with severe COVID‐19 by real‐time fluorescent RT‐PCR. In addition, much higher viral load in the bile juice than in the sputum indicated that a false positive error in the specimen was hardly possible. As far as we know, it is the first report about SARS‐CoV‐2 in the bile juice. We would love to share the information to other researchers. The report may have a significant impact on the clinical management and public health decision making. This article is protected by copyright. All rights reserved.


title: Comparison of the geographical distribution of feline immunodeficiency virus and feline leukemia virus infections in the United States of America (2000–2011)

text: BACKGROUND: Although feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) have similar risk factors and control measures, infection rates have been speculated to vary in geographic distribution over North America. Since both infections are endemic in North America, it was assumed as a working hypothesis that their geographic distributions were similar. Hence, the purpose of this exploratory analysis was to investigate the comparative geographical distribution of both viral infections. Counts of FIV (n=17,108) and FeLV (n=30,017) positive serology results (FIV antibody and FeLV ELISA) were obtained for 48 contiguous states and District of Columbia of the United States of America (US) from the IDEXX Laboratories website. The proportional morbidity ratio of FIV to FeLV infection was estimated for each administrative region and its geographic distribution pattern was visualized by a choropleth map. Statistical evidence of an excess in the proportional morbidity ratio from unity was assessed using the spatial scan test under the normal probability model. RESULTS: This study revealed distinct spatial distribution patterns in the proportional morbidity ratio suggesting the presence of one or more relevant and geographically varying risk factors. The disease map indicates that there is a higher prevalence of FIV infections in the southern and eastern US compared to FeLV. In contrast, FeLV infections were observed to be more frequent in the western US compared to FIV. The respective excess in proportional morbidity ratio was significant with respect to the spatial scan test (p < 0.05). CONCLUSIONS: The observed variability in the geographical distribution of the proportional morbidity ratio of FIV to FeLV may be related to the presence of an additional or unique, but yet unknown, spatial risk factor. Putative factors may be geographic variations in specific virus strains and rate of vaccination. Knowledge of these factors and the geographical distributions of these infections can inform recommendations for testing, management and prevention. However, further studies are required to investigate the potential association of these factors with FIV and FeLV.


title: Kidney involvement in COVID-19 and rationale for extracorporeal therapies

text: The prevalence of direct kidney involvement in novel coronavirus disease (COVID-19) is low, but such involvement is a marker of multiple organ dysfunction and severe disease. Here, we explore potential pathways of kidney damage and discuss the rationale for extracorporeal support with various blood purification strategies in patients who are critically ill with COVID-19.


title: Tales from the crypt and coral reef: the successes and challenges of identifying new herpesviruses using metagenomics

text: Herpesviruses are ubiquitous double-stranded DNA viruses infecting many animals, with the capacity to cause disease in both immunocompetent and immunocompromised hosts. Different herpesviruses have different cell tropisms, and have been detected in a diverse range of tissues and sample types. Metagenomics—encompassing viromics—analyses the nucleic acid of a tissue or other sample in an unbiased manner, making few or no prior assumptions about which viruses may be present in a sample. This approach has successfully discovered a number of novel herpesviruses. Furthermore, metagenomic analysis can identify herpesviruses with high degrees of sequence divergence from known herpesviruses and does not rely upon culturing large quantities of viral material. Metagenomics has had success in two areas of herpesvirus sequencing: firstly, the discovery of novel exogenous and endogenous herpesviruses in primates, bats and cnidarians; and secondly, in characterizing large areas of the genomes of herpesviruses previously only known from small fragments, revealing unexpected diversity. This review will discuss the successes and challenges of using metagenomics to identify novel herpesviruses, and future directions within the field.


title: Social status mediates the fitness costs of infection with canine distemper virus in Serengeti spotted hyenas

text: 1. The extent to which the fitness costs of infection are mediated by key life‐history traits such as age or social status is still unclear. Within populations, individual heterogeneity in the outcome of infection is the result of two successive processes; the degree of contact with the pathogen (exposure) and the immune response to infection. In social mammals, because individuals holding high social status typically interact more frequently with group members, they should be more often in contact with infected individuals than those of low social status. However, when access to resources is determined by social status, individuals with a high social status are often better nourished, have a greater opportunity to allocate resources to immune processes and therefore should have a smaller chance of succumbing to infection than individuals with low social status. 2. We investigated the risk and fitness costs of infection during a virulent epidemic of canine distemper virus (CDV) in a social carnivore, the spotted hyena, in the Serengeti National Park. We analysed two decades of detailed life‐history data from 625 females and 816 males using a multi‐event capture–mark–recapture model that accounts for uncertainty in the assignment of individual infection states. 3. Cubs of mothers with a high social status had a lower probability of CDV infection and were more likely to survive infection than those with low social status. Subadult and adult females with high social status had a higher infection probability than those with low social status. Subadult females and pre‐breeder males that had recovered from CDV infection had a lower survival than susceptible ones. 4. Our study disentangles the relative importance of individual exposure and resource allocation to immune processes, demonstrates fitness costs of infection for juveniles, particularly for those with low social status, shows that patterns of infection can be driven by different mechanisms among juveniles and adults and establishes a negative relationship between infection and fitness in a free‐ranging mammal. A http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.13059/suppinfo is available for this article.


title: Video as a public health knowledge transfer tool in Burkina Faso: A mixed evaluation comparing three narrative genres

text: BACKGROUND: The dengue virus is endemic in many low- and middle-income countries. In Burkina Faso, the proportion of fevers that could be due to dengue is growing. In 2013, a dengue epidemic spread there, followed by other seasonal outbreaks. Dengue is often confused with malaria, and health workers are not trained to distinguish between them. Three training videos using different narrative genres were tested with nursing students from two institutions in Ouagadougou: journalistic, dramatic and animated video. The study aimed to determine if video is an effective knowledge transfer tool, if narrative genre plays a role in knowledge acquisition, and which narrative elements are the most appreciated. METHODOLOGY: A mixed method research design was used. The relative effectiveness of the videos was verified through a quasi-experimental quantitative component with a comparison group and post-test measurements. A qualitative component identified participants’ perceptions regarding the three videos. Data were drawn from a knowledge test (n = 482), three focus groups with health professionals’ students (n = 46), and individual interviews with health professionals (n = 10). Descriptive statistics and single-factor variance analysis were produced. A thematic analysis was used to analyse qualitative data. PRINCIPAL FINDINGS: Results showed that all three videos led to significant rates of knowledge improvement when compared with the comparison group (p <0.05): 12.31% for the journalistic video, 20.58% for the dramatic video, and 18.91% for the animated video. The dramatic and animated videos produced a significantly higher increase in knowledge than did the journalistic video (with respectively 8.27% (p = 0.003) and 6.59% (p = 0.029) and can be considered equivalent with a difference of 1.68% (p = 0.895). Thematic analysis also revealed that these two videos were considered to be better knowledge transfer tools. Four key aspects are important to consider for a video to be effective: 1) transmitting information in a narrative form, 2) choosing good communicators, 3) creating a visual instrument that reinforces the message and 4) adapting the message to the local context. CONCLUSIONS: Video has proven to be an effective and appreciated knowledge transfer and training tool for health professionals, but the narrative genre of the videos can influence knowledge acquisition. The production of other videos should be considered for training or updating health professionals and their narrative genre taken into consideration. The actual context of constant circulation of new diseases, such as COVID-19, reaffirms the need to train health professionals.


title: Host-viral infection maps reveal signatures of severe COVID-19 patients

text: Viruses are a constant threat to global health as highlighted by the current COVID-19 pandemic. Currently, lack of data underlying how the human host interacts with viruses, including the SARS-CoV-2 virus, limits effective therapeutic intervention. We introduce Viral-Track, a computational method that globally scans unmapped scRNA-seq data for the presence of viral RNA, enabling transcriptional cell sorting of infected versus bystander cells. We demonstrate the sensitivity and specificity of Viral-Track to systematically detect viruses from multiple models of infection, including hepatitis B virus in an unsupervised manner. Applying Viral-Track to Bronchoalveloar-Lavage samples from severe and mild COVID-19 patients reveals a dramatic impact of the virus on the immune system of severe patients compared to mild cases. Viral-Track detects an unexpected co-infection of the human MetaPneumoVirus, present mainly in monocytes perturbed in type-I IFN-signaling. Viral-Track provides a robust technology for dissecting the mechanisms of viral-infection and pathology.


title: Interaction of paramyxoviruses with human basophils and their effect on histamine release

text: Abstract We have demonstrated that human peripheral blood basophils released histamine on direct incubation with paramyxoviruses in vitro. Most histamine release occurred during the first 15 to 30 minutes after challenge, depending on the dose of virus used; release initiated by virus was complete by 1 hour. At a virus/cell ratio of 1:1, Sendai virus caused 41 ± 9% histamine release, whereas parainfluenza type 3 (PI-3) virus caused 25 ± 5% release and respiratory syncytial (RS) virus caused 19 ± 5% release. Sendai, but not PI-3 or RS, also caused a decrease in cell number and release of lactic dehydrogenase; however, this apparent cell lysis did not account for all the histamine released. Incubation of cells with virus desensitized them to subsequent triggering by viruses but did not affect response of cells to other stimuli. Histamine release was dependent on the virus/cell ratio, temperature, and metabolic energy, but it was not strictly dependent on the presence of calcium in the extracellular medium. Histamine release was not affected by preincubation of cells with colchicine, suggesting that microtubules were not involved in the release process. Basophils desensitized by anti-IgE in the absence of calcium or treated with lactic acid to dissociate IgE molecules from membrane receptors released amounts of histamine similar to that of control basophils; thus, release was not initiated through Fcϵ receptors. It was found, however, that histamine release by these viruses was greatly reduced when concanavalin A was used for desensitization. These data demonstrate that the respiratory viruses studied can cause direct nonimmunologic release of histamine from human basophils. Our findings provide evidence for another mechanism by which respiratory viruses can initiate inflammation.


title: Accelerating the global response against the exponentially growing COVID-19 outbreak through decent data sharing

text: Abstract COVID-19 is a novel and exponentially-growing disease and consequently the accelerated development of knowledge from good data is possible quickly and globally. In order to combat the global pandemic of novel coronavirus disease 2019 (COVID-19), all humans on earth need to make difficult strategic decisions on three very different scales, all fueled by Analytical and Artificial Intelligence-based predictive Models (AAIMs).


Coronaviruses in children with febrile neutropenia


title: Asma infantil y del lactante

text: El asma es la enfermedad crónica más frecuente de la infancia. El diagnóstico suele ser fácil cuando se producen episodios de disnea espiratoria con sibilancias reversibles de forma espontánea o mediante el uso de broncodilatadores. Las radiografías de tórax, la exploración funcional respiratoria y el estudio alérgico constituyen el aspecto fundamental de las pruebas complementarias necesarias. El tratamiento de la crisis consiste en el uso de β2-adrenérgicos inhalados y, si es necesario, corticoides orales. El tratamiento de fondo tiene como objetivo limitar al máximo los síntomas y restaurar o mantener las funciones pulmonares normales. Se debe adaptar a la gravedad y al control de la enfermedad y los corticoides inhalados tienen un papel de elección.


title: Rhinosinusitis: Establishing definitions for clinical research and patient care

text: BACKGROUND: There is a need for more research on all forms of rhinosinusitis. Progress in this area has been hampered by a lack of consensus definitions and the limited number of published clinical trials. OBJECTIVES: To develop consensus definitions for rhinosinusitis and outline strategies useful in clinical trials. METHODS: Five national societies, The American Academy of Allergy, Asthma and Immunology; The American Academy of Otolaryngic Allergy; The American Academy of Otolaryngology Head and Neck Surgery; The American College of Allergy, Asthma and Immunology; and the American Rhinologic Society formed an expert panel from multiple disciplines. Over two days, the panel developed definitions for rhinosinusitis and outlined strategies for design of clinical trials. RESULTS: Committee members agreed to adopt the term “rhinosinusitis” and reached consensus on definitions and strategies for clinical research on acute presumed bacterial rhinosinusitis, chronic rhinosinusitis without polyposis, chronic rhinosinusitis with polyposis, and classic allergic fungal rhinosinusitis. Symptom and objective criteria, measures for monitoring research progress, and use of symptom scoring tools, quality-of-life instruments, radiologic studies, and rhinoscopic assessment were outlined for each condition. CONCLUSION: The recommendations from this conference should improve accuracy of clinical diagnosis and serve as a starting point for design of rhinosinusitis clinical trials.


title: The Effects of Simvastatin or Interferon-α on Infectivity of Human Norovirus Using a Gnotobiotic Pig Model for the Study of Antivirals

text: The lack of an animal model for human norovirus (HuNoV) has hindered the development of therapeutic strategies. This study demonstrated that a commonly used cholesterol-lowering statin medication, simvastatin, which increases HuNoV replication in an in vitro replicon system, also enhances HuNoV infectivity in the gnotobiotic (Gn) pig model. In contrast, oral treatment with interferon (IFN)-α reduces HuNoV infectivity. Young piglets, all with A or H1 histo-blood group antigens on enterocytes, were treated orally with 8 mg/kg/day of simvastatin; 5 days later, the pigs were inoculated orally with a GII.4 HuNoV (HS194/2009/US strain) and then treated with simvastatin for 5 more days. Simvastatin induced significantly earlier onset and longer duration of HuNoV fecal shedding in treated pigs, frequently with higher fecal viral titers. Simvastatin impaired poly (I:C)-induced IFN-α expression in macrophages or dendritic cells, possibly due to lowered toll-like receptor (TLR) 3 expression; however, the mechanisms were not related to interferon regulatory factor 3 or nuclear factor kappa B signaling pathway. Thus, the enhanced, earlier infectivity of HuNoV in simvastatin-treated pigs coincided with the inhibitory effect of simvastatin on innate immunity. In contrast to the increased HuNoV shedding that simvastatin induced, viral shedding during the treatment period was reduced or curtailed in the HuNoV-inoculated pigs pre-treated/treated with human IFN-α. Our findings are the first to indicate that IFN-α has potential as antiviral therapy against HuNoV. Based on these intriguing and novel findings using the Gn pig model, we confirmed that HuNoV infectivity is altered by treatment with simvastatin or IFN-α. Collectively, these findings indicate that Gn pigs are a useful model to test immunomodulators or efficacy of antivirals against HuNoV.


title: In vitro inhibition of severe acute respiratory syndrome coronavirus by chloroquine

text: Abstract We report on chloroquine, a 4-amino-quinoline, as an effective inhibitor of the replication of the severe acute respiratory syndrome coronavirus (SARS-CoV) in vitro. Chloroquine is a clinically approved drug effective against malaria. We tested chloroquine phosphate for its antiviral potential against SARS-CoV-induced cytopathicity in Vero E6 cell culture. Results indicate that the IC50 of chloroquine for antiviral activity (8.8±1.2μM) was significantly lower than its cytostatic activity; CC50 (261.3±14.5μM), yielding a selectivity index of 30. The IC50 of chloroquine for inhibition of SARS-CoV in vitro approximates the plasma concentrations of chloroquine reached during treatment of acute malaria. Addition of chloroquine to infected cultures could be delayed for up to 5h postinfection, without an important drop in antiviral activity. Chloroquine, an old antimalarial drug, may be considered for immediate use in the prevention and treatment of SARS-CoV infections.


The ethics of neuro-oncology in the era of COVID-19: lessons to be learned


docid: onyryfty

title: Gross and histopathological pulmonary findings in a COVID-19 associated death during self-isolation

text: Forensic investigations generally contain extensive morphological examinations to accurately diagnose the cause of death. Thus, the appearance of a new disease often creates emerging challenges in morphological examinations due to the lack of available data from autopsy- or biopsy-based research. Since late December 2019, an outbreak of a novel seventh coronavirus disease has been reported in China caused by “severe acute respiratory syndrome coronavirus 2” (SARS-CoV-2). On March 11, 2020, the new clinical condition COVID-19 (Corona-Virus-Disease-19) was declared a pandemic by the World Health Organization (WHO). Patients with COVID-19 mainly have a mild disease course, but severe disease onset might result in death due to proceeded lung injury with massive alveolar damage and progressive respiratory failure. However, the detailed mechanisms that cause organ injury still remain unclear. We investigated the morphological findings of a COVID-19 patient who died during self-isolation. Pathologic examination revealed massive bilateral alveolar damage, indicating early-phase “acute respiratory distress syndrome” (ARDS). This case emphasizes the possibility of a rapid severe disease onset in previously mild clinical condition and highlights the necessity of a complete autopsy to gain a better understanding of the pathophysiological changes in SARS-CoV-2 infections.


title: CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

text: The IFN-stimulated gene ubiquitin specific proteinase 18 (USP18) encodes a protein that negatively regulates T1 IFN signaling via stearic inhibition of JAK1 recruitment to the IFNα receptor 2 subunit (IFNAR2). Here we demonstrate that USP18 expression is induced by HIV-1 in a T1 IFN dependent manner. Experimental depletion of USP18 by CRISPR/Cas9 gene editing results in a significant restriction of HIV-1 replication in an induced pluripotent stem cell (iPSC)-derived macrophage model. In the absence of USP18, macrophages have increased responsiveness to stimulation with T1 IFNs with prolonged phosphorylation of STAT1 and STAT2 and increased expression of interferon-stimulated genes that are key for antiviral responses. Interestingly, HIV-1 requires some signaling through the T1 IFN receptor to replicate efficiently because a neutralizing antibody that inhibits T1 IFN activity reduces HIV-1 replication rate in monocyte-derived macrophages. USP18 induction by HIV-1 tunes the IFN response to optimal levels allowing for efficient transcription from the HIV-1 LTR promoter while minimizing the T1 IFN-induced antiviral response that would otherwise restrict viral replication and spread. Finally, iPSC and CRISPR/Cas9 gene targeting offer a powerful tool to study host factors that regulate innate immune responses.


title: The early impact of the Covid-19 pandemic on the global and Turkish economy

text: BACKGROUND/AIM: Individuals infected by the Covid-19 potentially are at risk of health and economic well-being. Today, the Covid-19 is a global issue, and the world economy can be interpreted as almost at the standstill. In this context, this study aims to discuss the potential first reactions of short and long term global economic impacts of the pandemic through sectors by assessing its costs according to the data announced for both the world and Turkey. In addition, this study tries to put forth possible economic and political scenarios for the post-pandemic world. MATERIALS AND METHODS: This is a review article that summarizes the current reports and discussions about the economic consequences of this historical event, and tries to make some inferences considering them. RESULTS: This pandemic has severe adverse effects on the employees, customers, supply chains and financial markets, in brief, most probably it will cause a global economic recession. Nevertheless, due to the uncertainty of the end of this pandemic, both the length and scale of this contraction are not predictable. CONCLUSION: It takes a while for the world economy to recover from the contraction. It seems that this pandemic will lead to a permanent shift in the world and its politics, especially in health, security, trade, employment, agriculture, manufacturing goods production and science policies. Since this new world might provide great opportunities for some countries that did not dominate world production before, governments should develop new strategies to adjust the new world order without much delay.


title: COVID-19 and the nervous system

text: A pandemic due to novel coronavirus arose in mid-December 2019 in Wuhan, China, and in 3 months’ time swept the world. The disease has been referred to as COVID-19, and the causative agent has been labelled SARS-CoV-2 due to its genetic similarities to the virus (SARS-CoV-1) responsible for the severe acute respiratory syndrome (SARS) epidemic nearly 20 years earlier. The spike proteins of both viruses dictate tissue tropism using the angiotensin-converting enzyme type 2 (ACE-2) receptor to bind to cells. The ACE-2 receptor can be found in nervous system tissue and endothelial cells among the tissues of many other organs. Neurological complications have been observed with COVID-19. Myalgia and headache are relatively common, but serious neurological disease appears to be rare. No part of the neuraxis is spared. The neurological disorders occurring with COVID-19 may have many pathophysiological underpinnings. Some appear to be the consequence of direct viral invasion of the nervous system tissue, others arise as a postviral autoimmune process, and still others are the result of metabolic and systemic complications due to the associated critical illness. This review addresses the preliminary observations regarding the neurological disorders reported with COVID-19 to date and describes some of the disorders that are anticipated from prior experience with similar coronaviruses.


title: An isolated epizootic of hemorrhagic-like fever in cats caused by a novel and highly virulent strain of feline calicivirus

text: Abstract An isolated epizootic of a highly fatal feline calicivirus (FCV) infection, manifested in its severest form by a systemic hemorrhagic-like fever, occurred over a 1-month period among six cats owned by two different employees and a client of a private veterinary practice. The infection may have started with an unowned shelter kitten that was hospitalized during this same period for a severe atypical upper respiratory infection. The causative agent was isolated from blood and nasal swabs from two cats; the electron microscopic appearance was typical for FCV and capsid gene sequencing showed it to be genetically similar to other less pathogenic field strains. An identical disease syndrome was recreated in laboratory cats through oral inoculation with tissue culture grown virus. During the course of transmission studies in experimental cats, the agent was inadvertently spread by caretakers to an adjoining room containing a group of four normal adult cats. One of the four older cats was found dead and a second was moribund within 48–72h in spite of symptomatic treatment; lesions in these animals were similar to those of the field cats but with the added feature of severe pancreatitis. The mortality in field cats, deliberately infected laboratory cats, and inadvertently infected laboratory cats ranged from 33–50%. This new isolate of calicivirus, named FCV-Ari, was neutralized at negligible to low titer by antiserum against the universal FCV-F9 vaccine strain. Cats orally immunized with FCV-F9, and then challenge-exposed shortly thereafter with FCV-Ari, developed a milder self-limiting form of disease, indicating partial protection. However, all of the field cats, including the three that died, had been previously immunized with parenteral FCV-F9 vaccine. FCV-Ari caused a disease that was reminiscent of Rabbit Hemorrhagic Disease, a highly fatal calicivirus infection of older rabbits.


title: The conserved stem-loop II structure at the 3' untranslated region of Japanese encephalitis virus genome is required for the formation of subgenomic flaviviral RNA

text: Flaviviruses accumulate abundant subgenomic RNA (sfRNA) in infected cells. It has been reported that sfRNA results from stalling of host 5’-to-3’ exoribonuclease XRN1 at the highly structured RNA of the 3’ untranslated region (UTR). Although XRN1 digestion of a 3’-terminal 800-nt RNA could stall at a position to generate the sfRNA in vitro, we found that knocking out XRN1 had no effect on the accumulation of sfRNA in Japanese encephalitis virus (JEV) infected cells. Mutagenesis studies revealed that the stemloop II (SLII) at the 3’ UTR is required for the accumulation of sfRNA. According to the results of an in vitro RNA-dependent RNA polymerase (RdRp) assay, the (-)10431-10566 RNA fragment, containing the putative promoter on the antigenome for the sfRNA transcription, binds to RdRp protein and exhibits a strong promoter activity. Taken together, our results indicate that the JEV sfRNA could be transcribed initially and then be trimmed by XRN1 or other unidentified exoribonucleases.


title: Application of the use of high‐throughput technologies to the determination of protein structures of bacterial and viral pathogens

text: The Structural Proteomics In Europe (SPINE) programme is aimed at the development and implementation of high‐throughput technologies for the efficient structure determination of proteins of biomedical importance, such as those of bacterial and viral pathogens linked to human health. Despite the challenging nature of some of these targets, 175 novel pathogen protein structures (∼220 including complexes) have been determined to date. Here the impact of several technologies on the structural determination of proteins from human pathogens is illustrated with selected examples, including the parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens.


title: Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris

text: The 3C-like protease (3CL(pro)) of severe acute respiratory syndrome associated coronavirus (SARS-CoV) is vital for SARS-CoV replication and is a promising drug target. Recombinant 3CL(pro) was expressed in Pichia pastoris GS115 as a 42 kDa protein that displayed a K (m) of 15 ± 2 μM with Dabcyl-KTSAVLQSGFRKME-Edans as substrate. Purified 3CL(pro) was used for inhibition and kinetic assays with seven flavonoid compounds. The IC(50) of six flavonoid compounds were 47–381 μM. Quercetin, epigallocatechin gallate and gallocatechin gallate (GCG) displayed good inhibition toward 3CL(pro) with IC(50) values of 73, 73 and 47 μM, respectively. GCG showed a competitive inhibition pattern with K (i) value of 25 ± 1.7 μM. In molecular docking experiments, GCG displayed a binding energy of −14 kcal mol(−1) to the active site of 3CL(pro) and the galloyl moiety at 3-OH position was required for 3CL(pro) inhibition activity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10529-011-0845-8) contains supplementary material, which is available to authorized users.


title: The book reopened on infectious diseases

text: Abstract Emerging infectious diseases represent a major challenge to human health worldwide. The risk of evolving new infectious pathogens has been intensifying due to urbanization, demographic changes, air travel, inappropriate use of antibiotics, and climate change. These pathogens can affect humans from urban centers to the remotest corners of the globe. Far from being a scourge of the past, infectious diseases are relevant for the world today.


title: Evaluation of the adverse effects of oral firocoxib in healthy dogs

text: This study evaluated the adverse effects of oral firocoxib in dogs. Six dogs (20.2 ± 6.3 kg) were studied. Values for complete blood count (CBC), serum urea, creatinine, alanine transaminase, alanine phosphatase, γ‐glutamyl transferase, occult blood in feces, platelet aggregation, and buccal mucosal bleeding time were measured before and 7, 14, 21, and 29 days after SID treatment with firocoxib 5.3 ± 0.34 mg/kg (FG) or lactose 1 mg/kg (LG) for 28 days, in a randomized crossover study. Gastrointestinal (GI) tract endoscopy was performed before treatment began and at 29 days. Lesions were scored from grade 0 to 6. Data were analyzed using anova and paired t‐tests (P < 0.05). None of the dogs presented adverse clinical effects. There were no significant changes in CBC, biochemical profiles within groups, or differences between groups. Pretreatment mean ± SD bleeding time (LG, 70.7 ± 32.1 sec; FG, 75.8 ± 38.1 sec) and platelet aggregation (LG, 86.4 ± 10.2%; FG, 85.6 ± 9.2%) were not significantly different from readings at 29 days (LG, 95.2 ± 25 sec; FG, 91.7 ± 24 sec and LG, 73.2 ± 15.1%; FG, 84 ± 10.3%) nor the groups were different. None of the dogs had positive fecal occult blood tests, and endoscopic lesion scores were grade 0 both before treatment and at 29 days. Administration of firocoxib did not cause any adverse effects on GI, or hematological or serum biochemical variables and appears to have been well tolerated by dogs.


title: Viral metacommunities associated to bats and rodents at different spatial scales

text: One of the main goals of community ecology is to measure the relative importance of environmental filters to understand patterns of species distribution at different temporal and spatial scales. Likewise, the identification of factors that shape symbiont metacommunity structures is important in disease ecology because resulting structures drive disease transmission. We tested the hypothesis that distributions of virus species and viral families from rodents and bats are defined by shared responses to host phylogeny and host functional characteristics, shaping the viral metacommunity structures at four spatial scales (Continental, Biogeographical, Zoogeographical, and Regional). The contribution of host phylogeny and host traits to the metacommunity of viruses at each spatial scale was calculated using a redundant analysis of canonical ordering (RDA). For rodents, at American Continental scale the coherence of viral species metacommunity increased while the spatial scale decreased and Quasi-Clementsian structures were observed. This pattern suggests a restricted distribution of viruses through their hosts, while in the Big Mass (Europe, Africa, and Asia), the coherence decreased as spatial scale decreased. Viral species metacommunities associated with bats was dominated by random structures along all spatial scales. We suggest that this random pattern is a result of the presence of viruses with high occupancy range such as rabies (73%) and coronavirus (27%), that disrupt such structures. At viral family scale, viral metacommunities associated with bats showed coherent structures, with the emergence of Quasi- Clementsian and Checkerboard structures. RDA analysis indicates that the assemblage of viral diversity associated with rodents and bats responds to phylogenetic and functional characteristics, which alternate between spatial scales. Several of these variations could be subject to the spatial scale, in spite of this, we could identify patterns at macro ecological scale. The application of metacommunity theory at symbiont scales is particularly useful for large-scale ecological analysis. Understanding the rules of host-virus association can be useful to take better decisions in epidemiological surveillance, control and even predictions of viral distribution and dissemination. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available for this article at 10.1556/168.2018.19.2.9 and is accessible for authorized users.


title: Infecciones por bocavirus humano en niños españoles: características clínicas y epidemiológicas de un virus respiratorio emergente

text: Introducción En el año 2005 se ha clonado un nuevo virus respiratorio, llamado bocavirus humano (HBoV) de muestras respiratorias procedentes de lactantes y niños suecos con infección respiratoria de vías bajas. Objetivos Determinar si HBoV ha circulado en España, estimar la frecuencia de infecciones por HBoV en los niños hospitalizados por infección respiratoria y describir sus características clínicas y epidemiológicas. Pacientes y métodos Estudio descriptivo, prospectivo de las infecciones confirmadas por HBoV en niños menores de 14 años, hospitalizados por infección respiratoria desde octubre de 2004 a junio de 2005. Para el diagnóstico virológico se realizó reacción en cadena de la polimerasa-transcripción inversa (RT-PCR) múltiple para virus respiratorio sincitial (VRS) A y B, influenza A, B y C, parainfluenza 1-4, adenovirus y rinovirus; PCR para metapneumovirus humano (hMPV) y PCR para HBoV en aspirado nasofaríngeo. Se describen las características clínicas y epidemiológicas de los pacientes. Resultados Se detectaron 52 casos de infección por HBoV, lo que supuso el 17,1 % (IC [intervalo de confianza] 95 %: 13 a 21) de los pacientes hospitalizados por procesos respiratorios. HBoV fue el tercer agente viral tras el VRS (30 %) y el rinovirus (25 %). En 39 casos (71,1 %) se detectó coinfección con otro virus respiratorio. El 50 % de los pacientes eran menores de 13,6 meses y el 75% menores de 2 años. Los diagnósticos más frecuentes fueron sibilancias recurrentes (55,8 %), bronquiolitis (21,2 %) y neumonía (15,4 %). Dos niños presentaron sepsis clínica con exantema petequial. El 71,2 % presentó fiebre superior a 38 °C e infiltrado radiológico el 44 %. Presentaron hipoxia el 55,8 % de los niños. Dos pacientes presentaron aislamientos positivos para HBoV en distintos episodios. Las coinfecciones fueron similares a las infecciones simples, excepto que presentaron hipoxia con más frecuencia, p=0,038. Conclusiones HBoV es uno de los virus más frecuentes en las infecciones respiratorias graves de los niños, sólo precedido por VRS y rinovirus. Las coinfecciones son muy frecuentes. La mayoría de los niños son lactantes con sibilancias recurrentes y bronquiolitis. Introduction In 2005 a new respiratory virus, called human bocavirus (HBoV), was cloned from respiratory samples from Swedish infants and children with lower respiratory tract infections. Objectives To determine whether HBoV has circulated in Spain, estimate the frequency of HBoV infections in patients hospitalized for respiratory infection and describe the clinical and epidemiological characteristics of these patients. Patients and methods We performed a descriptive prospective study of confirmed HBoV infections in patients aged < 14 years old, hospitalized for respiratory infections between October 2004 and June 2005. Virologic diagnosis was based on multiple RT-PCR for respiratory syncytial virus (RSV) A and B, influenza A,B, and C, parainfluenza 1-4, adenovirus and rhinovirus; PCR was used for human metapneumovirus (hMPV) and PCR in nasopharyngeal aspirates was used for HBoV. The clinical and epidemiological characteristics of patients were analyzed. Results Fifty-two cases of HBoV infection were detected, representing 17.1 % (95 % CI: 13 % a 21 %) of patients hospitalized for respiratory infections. HBoV was the third most frequent viral agent after RSV (30 %) and rhinovirus (25 %). In 39 patients (71.1 %) coinfection with another respiratory virus was detected. Fifty percent of the patients were aged less than 13.6 months and 75 % were aged less than 2 years. The most frequent diagnoses were recurrent wheezing (55.8 %), bronchiolitis (21.2 %) and pneumonia (15.4 %). Clinical sepsis with petechial exanthema was found in two patients. Fever > 38 °C was found in 72.1 % and radiological infiltrate in 44 %. Hypoxia was present in 55.8 % of the patients. HBoV was isolated in distinct episodes in two patients. Coinfections were similar to simple infections except that hypoxia was more frequent in the former (p 0.038). Conclusions HBoV is one of the most frequent viruses in severe respiratory infections in patients aged less than 14 years old. Only RSV and rhinovirus are more frequent. Coinfections are highly frequent. Most patients are infants with recurrent wheezing and bronchiolitis.


title: Seroepidemiological analysis of influenza pandemics in Shizuoka Prefecture and all Japan

text: Abstract (1) Seroepidemiological analysis of influenza pandemics (1986–2003) in Shizuoka Prefecture and all Japan revealed differences in geographical, annual, seasonal, and age distributions. (2) For 17 years, the pandemics generally began at the 50th week every year showing over 1.0 patient/clinic, reached the peak at 5th week the following year, and ended over 10–15th week. Two big A/H3N2 pandemics were seen in 1989/1990 and 1997/1998 seasons, claiming over 1 million patients in Japan. (3) As herald strains, A/H3N2 strains (A/Sydney-like) were found in October 1999, and B strains (B/Victoria- and B/Yamagata-like) were detected in July and November 1998 and, in August and December 2000 in Shizuoka. B/Shizuoka/1/98 strain was registered internationally as a vaccine-recommended strain. (4) A/H3N2 and B viruses were detected in 55–78% of flu patients (almost under 10 years) with encephalopathy in 1999/2000 and 78–91% in 2000/2001 by MDCK and reverse transcription polymerase chain reaction (RT-PCR) methods. (5) High hemagglutination inhibition (HI) titers over 40 in 250 persons were shown against A/Sydney/5/97 (H3N2), A/Yokohama/8/98 (H3N2), A/Panama/2007/99 (H3N2) and A/Moscow/10/99 (H1N1) strains, while low titers showed against A/Beijing/262/95 (H1N1) and A/New Caledonia/20/99 (H1N1), and B/Beijing/243/97, B/Shangdong/7/97 and B/Yamanashi/106/98 strains in 1998–2000. (6) In anti-HA titers against A/H3N2, A/H1N1 and B subtypes, clear generation gaps were observed between children (0–19 years), adults (20–59 years) and old men (over 60 years). (7) The pandemics are dependent on host immunity (acquired and vaccinated) and climatic conditions (low temperature, low humidity and limited rainfall), considering highly pathogenic avian influenza (HPAI) viruses (A/H5N1, A/H7N7) like severe acute respiratory syndrome (SARS) corona virus in 2002–2003.


title: Dynamic RNA Modifications in Posttranscriptional Regulation

text: Cellular RNAs can be chemically modified over a hundred different ways. These modifications were once thought to be static, discrete, and utilized to fine-tune RNA structure and function. However, recent studies have revealed that some modifications, like mRNA methylation, can be reversed, and these reversible modifications may play active roles in regulating diverse biological processes. In this perspective, we summarize examples of dynamic RNA modifications that affect biological functions. We further propose that reversible modifications might occur on tRNA, rRNA, and other noncoding RNAs to regulate gene expression analogous to the reversible mRNA methylation.


title: Western Washington State COVID-19 Experience: Keys to Flattening the Curve and Effective Health System Response

text: BACKGROUND: Washington State experienced the first major outbreak of COVID-19 in the United States and despite a significant number of cases, has seen a relatively low death rate per million population compared to other states with major outbreaks and has seen a substantial decrease in the projections for healthcare utilization, i.e. “flattening the curve”. This consensus report seeks to identify the key factors contributing to the effective health system disaster response in western WA. METHODS: A multidisciplinary, expert panel including individuals and organizations who were integral to managing the public health and emergency healthcare system response were engaged in a consensus process to identify the key themes and lessons learned and develop recommendations for ongoing management of the COVID-19 pandemic. RESULTS: Six key themes were identified including: early communication and coordination among stakeholders; regional coordination of the healthcare system response; rapid development and access to viral testing; proactive management of long-term care & skilled nursing facilities; proactive management of vulnerable populations; and effective physical distancing in the community. CONCLUSIONS: Based on the lessons learned in each of the areas identified by the panel, 11 recommendations are provided to support the healthcare system disaster response in managing future outbreaks.


Neuro-Ophthalmology in the Era of COVID-19: Future Implications of a Public Health Crisis


title: The Use of Non‐invasive Vagus Nerve Stimulation to Treat Respiratory Symptoms Associated With COVID‐19: A Theoretical Hypothesis and Early Clinical Experience

text: OBJECTIVES: Coronavirus disease 2019 (COVID‐19), caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), is a pandemic with no specific therapeutic agents and substantial mortality, and finding new treatments is critical. Most cases are mild, but a significant minority of patients develop moderate to severe respiratory symptoms, with the most severe cases requiring intensive care and/or ventilator support. This respiratory compromise appears to be due to a hyperimmune reaction, often called a cytokine storm. Vagus nerve stimulation has been demonstrated to block production of cytokines in sepsis and other medical conditions. We hypothesize that non‐invasive vagus nerve stimulation (nVNS) might provide clinical benefits in patients with respiratory symptoms similar to those associated with COVID‐19. MATERIALS AND METHODS: Information on two case reports was obtained via email correspondence and phone interviews with the patients. RESULTS: Both patients reported clinically meaningful benefits from nVNS therapy. In case 1, the patient used nVNS to expedite symptomatic recovery at home after hospital discharge and was able to discontinue use of opioid and cough suppressant medications. In case 2, the patient experienced immediate and consistent relief from symptoms of chest tightness and shortness of breath, as well as an improved ability to clear his lungs. CONCLUSIONS: Preliminary observations and a strong scientific foundation suggest that nVNS might provide clinical benefits in patients with COVID‐19 via multiple mechanisms.


Was SARS a mental health catastrophe?


title: Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China

text: Six porcine epidemic diarrhea viruses (PEDVs) were isolated from the fecal samples of piglets infected with PEDV in 2006 in China. The membrane (M) protein genes of six PEDV isolates were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), then cloned, sequenced, and compared with each other as well as those ten PEDV reference strains. The M protein genes of six Chinese PEDV isolates consisted of 692 nucleotides containing a single open reading frame (ORF) of 681 nucleotides, which encoded a 226aa-long peptide. The conserved intergenic motif (ATAAAC), as previously recognized in Br1/87, was found in the 5 nucleotides upstream of the initiator ATG of M protein genes of six Chinese PEDV isolates. The hexamer motif was also found in CV777, JMe2, LZC, and QH. The M protein of six isolates had three main transmembrane domains (aa20–38, aa43–65, aa75–97). The M protein of one isolate, CH/IMT/06, had one potential glycosylation site, but those of the other five isolates had two. The glycosylation sequence Asn-Phe-Thr was highly conserved in the M proteins of six PEDV isolates. The six PEDV isolates showed nucleotide sequence homology between 98.8 and 100% and deduced amino acid sequence homology between 98.2 and 100% with each other. The nucleotide and amino acid identity of M protein genes between the six PEDV isolates and ten reference PEDV strains varied from 97.2 to 99.4% and 96.9 to 100%, respectively. On the basis of the phylogenetic relationship of M protein genes, six Chinese PEDV isolates composed of a separate cluster including one Chinese strain JS-2004-02, however, not including the Chinese strain LJB/03. These results demonstrated that there was a new genotype of PEDV prevailing in China.


Close Relative of Human Middle East Respiratory Syndrome Coronavirus in Bat, South Africa


title: The CoV-2 outbreak: how hematologists could help to fight Covid-19

text: Abstract COVID-19 is a medical emergency, with 20 % of patients presenting with severe clinical manifestations. From the pathogenetic point of view, COVID-19 mimics two other well-known diseases characterized by cytokine storm and hyper-activation of the immune response, with consequent organ damage: acute graft-versus-host disease (aGVHD) and macrophage activation syndrome (MAS). Hematologists are confident with these situations requiring a prompt therapeutic approach for switching off the uncontrolled cytokine release; here, we discuss pros and cons of drugs that are already employed in hematology in the light of their possible application in COVID-19. The most promising drugs might be: Ruxolitinib, a JAK1/2 inhibitor, with a rapid and powerful anti-cytokine effect, tyrosine kinase inhibitors (TKIs), with their good anti-inflammatory properties, and perhaps the anti-Cd26 antibody Begelomab. We also present immunological data from gene expression experiments where TKIs resulted effective anti-inflammatory and pro-immune drugs. A possible combined treatment algorithm for COVID-19 is here proposed.


title: Coronavirus occurrence and transmission over 8 years in the HIVE cohort of households in Michigan

text: BACKGROUND: As part of the Household Influenza Vaccine Evaluation (HIVE) study, acute respiratory infections (ARI) have been identified in children and adults over 8 years. METHODS: Annually, 890 to 1441 individuals were followed and contacted weekly to report ARIs. Specimens collected during illness were tested for human coronaviruses (HCoV) types OC43, 229E, HKU1, and NL63. RESULTS: In total, 993 HCoV infections were identified over 8 years, with OC43 most commonly seen and 229E the least. HCoVs were detected in a limited time period, between December and April/May, and peaked in January/February. Highest infection frequency was in children <5 years (18 per 100 person-years), with little variation in older age groups (range: 7 to 11 per 100 person-years). Overall, 9% of adult cases and 20% of cases in children were associated with medical consultation. Of the 993 infections, 260 were acquired from an infected household contact. The serial interval between index and household-acquired cases ranged from 3.2 to 3.6 days and the secondary infection risk ranged from 7.2% to 12.6% by type. CONCLUSIONS: Coronaviruses are sharply seasonal. They appear, based on serial interval and secondary infection risk, to have similar transmission potential to influenza A(H3N2) in the same population.


Part 1: Executive summary 2010 International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations


title: Crystal Structure and Functional Analysis of the SARS-Coronavirus RNA Cap 2′-O-Methyltransferase nsp10/nsp16 Complex

text: Cellular and viral S-adenosylmethionine-dependent methyltransferases are involved in many regulated processes such as metabolism, detoxification, signal transduction, chromatin remodeling, nucleic acid processing, and mRNA capping. The Severe Acute Respiratory Syndrome coronavirus nsp16 protein is a S-adenosylmethionine-dependent (nucleoside-2′-O)-methyltransferase only active in the presence of its activating partner nsp10. We report the nsp10/nsp16 complex structure at 2.0 Å resolution, which shows nsp10 bound to nsp16 through a ∼930 Å(2) surface area in nsp10. Functional assays identify key residues involved in nsp10/nsp16 association, and in RNA binding or catalysis, the latter likely through a SN2-like mechanism. We present two other crystal structures, the inhibitor Sinefungin bound in the S-adenosylmethionine binding pocket and the tighter complex nsp10(Y96F)/nsp16, providing the first structural insight into the regulation of RNA capping enzymes in (+)RNA viruses.


title: The host immune response in respiratory virus infection: balancing virus clearance and immunopathology

text: The respiratory tract is constantly exposed to the external environment, and therefore, must be equipped to respond to and eliminate pathogens. Viral clearance and resolution of infection requires a complex, multi-faceted response initiated by resident respiratory tract cells and innate immune cells and ultimately resolved by adaptive immune cells. Although an effective immune response to eliminate viral pathogens is essential, a prolonged or exaggerated response can damage the respiratory tract. Immune-mediated pulmonary damage is manifested clinically in a variety of ways depending on location and extent of injury. Thus, the antiviral immune response represents a balancing act between the elimination of virus and immune-mediated pulmonary injury. In this review, we highlight major components of the host response to acute viral infection and their role in contributing to mitigating respiratory damage. We also briefly describe common clinical manifestations of respiratory viral infection and morphological correlates. The continuing threat posed by pandemic influenza as well as the emergence of novel respiratory viruses also capable of producing severe acute lung injury such as SARS-CoV, MERS-CoV, and enterovirus D68, highlights the need for an understanding of the immune mechanisms that contribute to virus elimination and immune-mediated injury.


title: Operational efficiency and sustainability of vector control of malaria and dengue: descriptive case studies from the Philippines

text: BACKGROUND: Analysis is lacking on the management of vector control systems in disease-endemic countries with respect to the efficiency and sustainability of operations. METHODS: Three locations were selected, at the scale of province, municipality and barangay (i.e. village). Data on disease incidence, programme activities, and programme management were collected on-site through meetings and focus group discussions. RESULTS: Adaptation of disease control strategies to the epidemiological situation per barangay, through micro-stratification, brings gains in efficiency, but should be accompanied by further capacity building on local situational analysis for better selection and targeting of vector control interventions within the barangay. An integrated approach to vector control, aiming to improve the rational use of resources, was evident with a multi-disease strategy for detection and response, and by the use of combinations of vector control methods. Collaboration within the health sector was apparent from the involvement of barangay health workers, re-orientation of job descriptions and the creation of a disease surveillance unit. The engagement of barangay leaders and use of existing community structures helped mobilize local resources and voluntary services for vector control. In one location, local authorities and the community were involved in the planning, implementation and evaluation of malaria control, which triggered local programme ownership. CONCLUSIONS: Strategies that contributed to an improved efficiency and sustainability of vector control operations were: micro-stratification, integration of vector control within the health sector, a multi-disease approach, involvement of local authorities, and empowerment of communities. Capacity building on situational analysis and vector surveillance should be addressed through national policy and guidelines.


docid: zonfottc

title: Application of virus-like particles (VLP) to NMR characterization of viral membrane protein interactions

text: The membrane proteins of viruses play critical roles in the virus life cycle and are attractive targets for therapeutic intervention. Virus-like particles (VLP) present the possibility to study the biochemical and biophysical properties of viral membrane proteins in their native environment. Specifically, the VLP constructs contain the entire protein sequence and are comprised of native membrane components including lipids, cholesterol, carbohydrates and cellular proteins. In this study we prepare VLP containing full-length hemagglutinin (HA) or neuraminidase (NA) from influenza and characterize their interactions with small molecule inhibitors. Using HA-VLP, we first show that VLP samples prepared using the standard sucrose gradient purification scheme contain significant amounts of serum proteins, which exhibit high potential for non-specific interactions, thereby complicating NMR studies of ligand-target interactions. We then show that the serum contaminants may be largely removed with the addition of a gel filtration chromatography step. Next, using HA-VLP we demonstrate that WaterLOGSY NMR is significantly more sensitive than Saturation Transfer Difference (STD) NMR for the study of ligand interactions with membrane bound targets. In addition, we compare the ligand orientation to HA embedded in VLP with that of recombinant HA by STD NMR. In a subsequent step, using NA-VLP we characterize the kinetic and binding properties of substrate analogs and inhibitors of NA, including study of the H274Y-NA mutant, which leads to wide spread resistance to current influenza antivirals. In summary, our work suggests that VLP have high potential to become standard tools in biochemical and biophysical studies of viral membrane proteins, particularly when VLP are highly purified and combined with control VLP containing native membrane proteins.


docid: pyfbzyxp

title: Virus–Host Interactions Involved in Lassa Virus Entry and Genome Replication

text: Lassa virus (LASV) is the causative agent of Lassa fever, a human hemorrhagic disease associated with high mortality and morbidity rates, particularly prevalent in West Africa. Over the past few years, a significant amount of novel information has been provided on cellular factors that are determinant elements playing a role in arenavirus multiplication. In this review, we focus on host proteins that intersect with the initial steps of the LASV replication cycle: virus entry and genome replication. A better understanding of relevant virus–host interactions essential for sustaining these critical steps may help to identify possible targets for the rational design of novel therapeutic approaches against LASV and other arenaviruses that cause severe human disease.


title: The role of chemokines and chemokine receptors in multiple sclerosis

text: Abstract Multiple sclerosis (MS) is a chronic inflammatory disease that is characterized by leukocyte infiltration and subsequent axonal damage, demyelinating inflammation, and formation of sclerosing plaques in brain tissue. The results of various studies in patients indicate that autoimmunity and inflammation make an important impact on the pathogenesis of MS. Chemokines are key mediators of inflammation development and cell migration, mediating various immune cell responses, including chemotaxis and immune activation, and are important in immunity and inflammation, therefore we focus on chemokines and their receptors in multiple sclerosis. In this article, we summarize the study of the role of prominent chemokines and their receptors in MS patients and MS animal modelsand discuss their potential significance in inflammatory injury and repair of MS. We have also summarized the progress in the treatment of multiple sclerosis antagonists in recent years with chemokine receptors as targets.


title: High prevalence and genetic diversity of hepatitis B viruses in insectivorous bats from China

text: Abstract Bats have been identified as the hosts of hepatitis B virus (HBV) in recent years and bats HBV can infect human hepatocyte. We investigated the prevalence and genetic diversity of HBV in bats in China. In this study, a total of 197 insectivorous bats belonging to 10 bat species were captured from karst caves in Mengyin County, Shandong Province and Xianning City, Hubei Province, China. PCR amplification indicated that in total 6.6% (13/197) bats were positive to HBVs. The HBV positive rate in bats was 7.1% (9/127) and 5.7% (4/70) in Shandong Province and Hubei Province, respectively. Phylogenetic analysis indicated that HBV from the two places were in the same cluster with 90.5%–99.5% homology, but distinct from bat HBVs from other places in China and other countries. We concluded that HBV was prevalent and genetic diversified in bats, supporting the hypothesis that bats may be the origin of primate hepadnaviruses.


title: Viral replicons as valuable tools for drug discovery

text: RNA viruses can cause severe diseases such as dengue, Lassa, chikungunya and Ebola. Many of these viruses can only be propagated under high containment levels, necessitating the development of low containment surrogate systems such as subgenomic replicons and minigenome systems. Replicons are self-amplifying recombinant RNA molecules expressing proteins sufficient for their own replication but which do not produce infectious virions. Replicons can persist in cells and are passed on during cell division, enabling quick, efficient and high-throughput testing of drug candidates that act on viral transcription, translation and replication. This review will explore the history and potential for drug discovery of hepatitis C virus, dengue virus, respiratory syncytial virus, Ebola virus and norovirus replicon and minigenome systems.


title: The future of manufacturing: a Delphi-based scenario analysis on Industry 4.0

text: Abstract Industry 4.0 is expected to impart profound changes to the configuration of manufacturing companies with regards to what their value proposition will be and how their production network, supplier base and customer interfaces will develop. The literature on the topic is still fragmented; the features of the emerging paradigm appear to be a contested territory among different academic disciplines. This study assumes a value chain perspective to analyze the evolutionary trajectories of manufacturing companies. We developed a Delphi-based scenario analysis involving 76 experts from academia and practice. The results highlight the most common expectations as well as controversial issues in terms of emerging business models, size, barriers to entry, vertical integration, rent distribution, and geographical location of activities. Eight scenarios provide a concise outlook on the range of possible futures. These scenarios are based on four main drivers which stem from the experts’ comments: demand characteristics, transparency of data among value chain participants, maturity of additive manufacturing and advanced robotics, and penetration of smart products. Researchers can derive from our study a series of hypotheses and opportunities for future research on Industry 4.0. Managers and policymakers can leverage the scenarios in long-term strategic planning.


title: Key Ethical Issues Discussed at CDC-Sponsored International, Regional Meetings to Explore Cultural Perspectives and Contexts on Pandemic Influenza Preparedness and Response

text: Background: Recognizing the importance of having a broad exploration of how cultural perspectives may shape thinking about ethical considerations, the Centers for Disease Control and Prevention (CDC) funded four regional meetings in Africa, Asia, Latin America, and the Eastern Mediterranean to explore these perspectives relevant to pandemic influenza preparedness and response. The meetings were attended by 168 health professionals, scientists, academics, ethicists, religious leaders, and other community members representing 40 countries in these regions. Methods: We reviewed the meeting reports, notes and stories and mapped outcomes to the key ethical challenges for pandemic influenza response described in the World Health Organization’s (WHO’s) guidance, Ethical Considerations in Developing a Public Health Response to Pandemic Influenza: transparency and public engagement, allocation of resources, social distancing, obligations to and of healthcare workers, and international collaboration. Results: The important role of transparency and public engagement were widely accepted among participants. However, there was general agreement that no "one size fits all" approach to allocating resources can address the variety of economic, cultural and other contextual factors that must be taken into account. The importance of social distancing as a tool to limit disease transmission was also recognized, but the difficulties associated with this measure were acknowledged. There was agreement that healthcare workers often have competing obligations and that government has a responsibility to assist healthcare workers in doing their job by providing appropriate training and equipment. Finally, there was agreement about the importance of international collaboration for combating global health threats. Conclusion: Although some cultural differences in the values that frame pandemic preparedness and response efforts were observed, participants generally agreed on the key ethical principles discussed in the WHO’s guidance. Most significantly the input gathered from these regional meetings pointed to the important role that procedural ethics can play in bringing people and countries together to respond to the shared health threat posed by a pandemic influenza despite the existence of cultural differences.


title: Decontamination of face masks with steam for mask reuse in fighting the pandemic COVID‐19: experimental supports

text: The COVID‐19 pandemic caused by the novel coronavirus SARS‐CoV‐2 has claimed many lives worldwide. Wearing medical masks or N95 masks (namely N95 respirators) can slow the virus spread and reduce the infection risk. Reuse of these masks can minimize waste, protect the environment, and help to solve the current imminent shortage of masks. Disinfection of used masks is needed for reuse of them with safety, but improper decontamination can damage the blocking structure of masks. In this study, we demonstrated, using avian coronavirus of infectious bronchitis virus to mimic SARS‐CoV‐2, that medical masks and N95 masks remained their blocking efficacy after being steamed on boiling water even for 2 hours. We also demonstrated that three brands of medical masks blocked over 99% viruses in aerosols. The avian coronavirus was completely inactivated after being steamed for 5 minutes. Together, this study suggested that medical masks are adequate for use on most social occasions, and both medical masks and N95 masks can be reused for a few days with steam decontamination between use. This article is protected by copyright. All rights reserved.


title: Clinical characteristics of 3,062 COVID‐19 patients: a meta‐analysis

text: OBJECTIVE: We aim to systematically review the clinical characteristics of Coronavirus disease 2019 (COVID‐19). METHODS: Seven datebases were searched to collect studies about the clinical characteristics of COVID‐19 from 1 January 2020 to 28 February 2020. Then, meta‐analysis was performed by using Stata12.0 software. RESULTS: A total of 38 studies involving 3 062 COVID‐19 patients were included. Meta‐analysis showed that a higher proportion of infected patients were male (56.9%). The incidence rate of respiratory failure or ARDS was 19.5% and the fatality rate was 5.5%. Fever (80.4%), fatigue (46%), cough (63.1%) and expectoration (41.8%) were the most common clinical manifestations. Other common symptoms included muscle soreness (33%), anorexia (38.8%), chest tightness (35.7%), shortness of breath (35%), dyspnea (33.9%). Minor symptoms included nausea and vomiting (10.2%), diarrhea (12.9%), headache (15.4%), pharyngalgia(13.1%), shivering (10.9%) and abdominal pain (4.4%). Patients with asymptomatic was 11.9%. Normal leukocytes counts (69.7%), lymphopenia (56.5%), elevated C‐reactive protein levels (73.6%), elevated ESR (65.6%) and oxygenation index decreased (63.6%) were observed in most patients. About 37.2% of patients with elevated D‐dimer, 25.9% of patients with leukopenia, along with abnormal levels of liver function (29%) and renal function (25.5%). Other findings included leukocytosis (12.6%) and elevated procalcitonin (17.5%). Only 25.8% of patients had lesions involving single lung and 75.7% of patients had lesions involving bilateral lungs. CONCLUSIONS: The most commonly experienced symptoms of COVID‐19 patients were fever, fatigue, cough and expectoration. A relatively small percentage of patients were asymptomatic. Most patients showed normal leucocytes counts, lymphopenia, elevated levels of C‐reactive protein and ESR. Bilateral lungs involvement was common. This article is protected by copyright. All rights reserved.


title: Dietary Management of Food Protein-Induced Enterocolitis Syndrome during COVID-19 Pandemic

text: As communities struggle to adapt to life under the threat of the global pandemic, COVID-19, those living with Food Protein-Induced Enterocolitis Syndrome (FPIES) must adapt with additional difficulties. Social distancing and shelter-in-place strategies have been implemented, resulting in fewer supermarkets trips, stockpile-purchasing behaviors in up to 74.5% of those surveyed(1), and shortages of staple food items all with potential impact on the availability of foods for those on limited diets. Concern about allergic reactions make exploring alternative or new ingredients undesirable or untenable. Remaining safe at home is important to avoid trips to the emergency department where families may be exposed to the COVID-19 virus and medical attention can be limited due to the burden on global health systems. Parents of children with FPIES are also understandably concerned about meeting their child’s nutritional needs during these times of sheltering-in-place. Now more than ever, advice on what foods to serve and when to serve them is critically important.


title: 19 treatment regimens?

text: Abstract The pandemic of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a significant threat to global health. Currently, no specific prophylactic and therapeutic treatment is available. No evidence from randomized clinical trials (RCTs) that a treatment may ameliorate the clinical outcome of patients with COVID-19 exists with the only exception of preliminary evidence from remdesivir trials. Here, we present evidence from the literature and a compelling hypothesis on the potential immunomodulatory, iron chelating and anti-oxidant effects of iron chelators in the treatment of COVID-19 and its complications. Interestingly, iron chelation has been shown in vitro to suppress endothelial inflammation in viral infection, which is the main pathophysiologic mechanism behind systemic organ involvement induced by SARS-CoV-2, by inhibiting IL-6 synthesis through decreasing NF-kB. Iron chelators exhibit iron chelating, antiviral and immunomodulatory effects in vitro and in vivo, particularly against RNA viruses. These agents could attenuate ARDS and help control SARS-CoV-2 via multiple mechanisms including: 1) inhibition of viral replication; 2) decrease of iron availability; 3) upregulation of B cells; 4) improvement of the neutralizing anti-viral antibody titer; 5) inhibition of endothelial inflammation and 6) prevention of pulmonary fibrosis and lung decline via reduction of pulmonary iron accumulation. Both retrospective analyses of data in electronic health records, as well as proof of concept studies in humans and large RCTs are needed to fully elucidate the efficacy and safety of iron chelating agents in the therapeutic armamentarium of COVID-19, probably as an adjunctive treatment.


docid: imrjlhia

title: Principi e indicazioni dell’assistenza circolatoria e respiratoria extracorporea in chirurgia toracica

text: In origine, l’extracorporeal membrane oxygenation (ECMO) era una tecnica di assistenza respiratoria che utilizzava uno scambiatore gassoso a membrana. Per estensione, l’ECMO è diventata una tecnica respiratoria e cardiopolmonare utilizzata in caso di deficit respiratorio e/o cardiaco nell’attesa della restaurazione della funzione deficitaria o di un eventuale trapianto. Il supporto emodinamico può essere parziale o totale. Gli accessi vascolari possono essere periferici o centrali. Questo tipo di assistenza utilizza il concetto di circolazione extracorporea (CEC) sanguigna che in epoca moderna si è estesa con l’utilizzo di polmoni artificiali a membrana. Il circuito di base è semplice e comprende una pompa, un ossigenatore (che permette al sangue di caricarsi di O2 e di eliminare CO2) e delle vie d’accesso (una di drenaggio e una di reinfusione). La sua attuazione è facile, veloce e può essere avviata al letto del malato. Il miglioramento delle attrezzature, una migliore conoscenza delle tecniche e delle indicazioni, e le politiche di salute pubblica hanno reso popolare questa tecnica. Alcuni centri di chirurgia toracica la utilizzano di routine come assistenza alla realizzazione di un intervento terapeutico (soprattutto trapianto) assieme a team di rianimazione per il trattamento della sindrome da distress respiratorio acuto. Nel quadro della malattia polmonare dell’adulto, l’idea principale è quella di sviluppare il concetto di strategia minimalista con l’uso di una CEC adiuvante parziale – più che sostitutiva totale – che permetterebbe il recupero metabolico ad integrum del paziente. Nei prossimi anni, i progressi della tecnologia e dell’ingegneria così come le conoscenze approfondite permetteranno il miglioramento della prognosi dei pazienti colpiti da deficit respiratorio sotto assistenza meccanica.


title: Application of a portable instrument for rapid and reliable detection of SARS‐CoV‐2 infection in any environment

text: The ongoing outbreak of the novel coronavirus (SARS‐CoV‐2) infection is creating serious challenges for health laboratories that seek to identify viral infections as early as possible, optimally at the earliest appearance of symptom. Indeed, there is urgent need to develop and deploy robust diagnostic methodologies not only to use in health laboratory environments but also directly in places where humans circulate and spread the virus such as airports, trains, boats, and any public aggregation places. The success of a reliable and sensitive asymptomatic diagnosis relies on the identification and measurement of informative biomarkers from human host and virus in a rapid, sensitive, and inexpensive manner. The objective of this article is to describe an innovative multidisciplinary approach to develop an efficient, inexpensive, and easy‐to‐use portable instrument (bCUBE(®) by Hyris Ltd) that can be employed as a surveillance system for the emergency caused by SARS‐CoV‐2. A solution for Coronavirus testing, compliant with CDC guidelines, is scheduled to be released in the next weeks. In addition, we will describe a workflow and path of an integrated multi‐omic approach that will lead to host and pathogen biomarker discovery in order to train the instrument to provide reliable results based on a specific biomarker's fingerprint of SARS‐CoV‐2 infection.


title: HexaPrime: A novel method for detection of coronaviruses

text: Abstract Despite intense efforts to develop novel and better tools to identify known viruses and to discover new viruses, establishing etiological roles for viruses in human disease is challenging. In large part, this may be attributed to the high variability of viral species and the difficulties in developing broad-spectrum, yet specific, diagnostic assays. To overcome this problem, a novel method for the detection of viruses is described in the current manuscript. The technique relies on the addition of synthetic oligonucleotides to both termini of RNA fragments in a sequence-dependent manner during first- and second-strand DNA synthesis; these oligonucleotides are used subsequently for amplification of the viral nucleic acids of interest. The recognition of the target sequence by the oligonucleotides is mediated by short (6–8nt) conserved regions, which facilitates development of broad–spectrum assays. The method has been tested for coronaviruses, although it may be also adopted for other RNA viruses.


title: Genome sequencing of the first SARS-CoV-2 reported from patients with COVID-19 in Ecuador

text: SARS-CoV-2, the etiological agent of COVID-19 was first described in Wuhan in December 2019 and has now spread globally. Ecuador was the second country in South America to report confirmed cases. The first case reported in Quito, the capital city of Ecuador, was a tourist who came from the Netherlands and presented symptoms on March 10th, 2020 (index case). In this work we used the MinION platform (Oxford Nanopore Technologies) to sequence the metagenome of the bronchoalveolar lavage (BAL) from this case reported, and subsequently we sequenced the whole genome of the index case and other three patients using the ARTIC network protocols. Our data from the metagenomic approach confirmed the presence of SARS-CoV-2 coexisting with pathogenic bacteria suggesting coinfection. Relevant bacteria found in the BAL metagenome were Streptococcus pneumoniae, Mycobacterium tuberculosis, Staphylococcus aureus and Chlamydia spp. Lineage assignment of the four whole genomes revealed three different origins. The variant HEE-01 was imported from the Netherlands and was assigned to B lineage, HGSQ-USFQ-018, belongs to the B.1 lineage showing nine nucleotide differences with the reference strain and grouped with sequences from the United Kingdom, and HGSQ-USFQ-007 and HGSQ-USFQ-010 belong to the B lineage and grouped with sequences from Scotland. All genomes show mutations in their genomes compared to the reference strain, which could be important to understand the virulence, severity and transmissibility of the virus. Our findings also suggest that there were at least three independent introductions of SARS-CoV-2 to Ecuador.


Suggestions from Cremona, Italy - two months into the pandemic at the frontline of COVID-19 in Europe


title: An unfolded protein-induced conformational switch activates mammalian IRE1

text: The unfolded protein response (UPR) adjusts the cell’s protein folding capacity in the endoplasmic reticulum (ER) according to need. IRE1 is the most conserved UPR sensor in eukaryotic cells. It has remained controversial, however, whether mammalian and yeast IRE1 use a common mechanism for ER stress sensing. Here, we show that similar to yeast, human IRE1α’s ER-lumenal domain (hIRE1α LD) binds peptides with a characteristic amino acid bias. Peptides and unfolded proteins bind to hIRE1α LD’s MHC-like groove and induce allosteric changes that lead to its oligomerization. Mutation of a hydrophobic patch at the oligomerization interface decoupled peptide binding to hIRE1α LD from its oligomerization, yet retained peptide-induced allosteric coupling within the domain. Importantly, impairing oligomerization of hIRE1α LD abolished IRE1’s activity in living cells. Our results provide evidence for a unifying mechanism of IRE1 activation that relies on unfolded protein binding-induced oligomerization.


title: Sample descriptors linked to metagenomic sequencing data from human and animal enteric samples from Vietnam

text: There is still limited information on the diversity of viruses co-circulating in humans and animals. Here, we report data obtained from a large field collection of enteric samples taken from humans, pigs, rodents and other mammal hosts in Vietnam between 2012 and 2016. Each of 2100 stool or rectal swab samples was subjected to virally-enriched agnostic metagenomic sequencing; the short read sequence data are accessible from the European Nucleotide Archive (ENA). We link the sequence data to metadata on host type and demography and geographic location, distinguishing hospital patients, members of a cohort identified as a high risk of zoonotic infections (e.g. abattoir workers, rat traders) and animals. These data are suitable for further studies of virus diversity and virus discovery in humans and animals from Vietnam and to identify viruses found in multiple hosts that are potentially zoonotic.


title: Clinical Characteristics of Acute Lower Extremity Deep Venous Thrombosis Diagnosed by Duplex in Patients Hospitalized for Coronavirus Disease (COVID-19)

text: OBJECTIVE: Little is known about coronavirus disease (COVID-19)-associated hypercoagulability. We sought to characterize patients with deep venous thrombosis (DVT) identified after admission for COVID-19. METHODS: All adult patients admitted to Montefiore Medical Center (MMC) from March 1, 2020 to April 10, 2020 and undergoing lower extremity venous duplex for DVT evaluation were included. Patients admitted with suspicion of COVID-19 were divided into severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positive and SARS-CoV-2 negative groups based on in-hospital test results. Patients without clinical suspicion for COVID-19 were not tested. A retrospective case-control study design was used to identify potential risk factors for DVT in patients with COVID-19. Demographic, radiographic and laboratory values were abstracted and analyzed. RESULTS: During the study period, 3,404 patients with confirmed COVID-19 were admitted to the hospital. Of the 135 SARS-CoV-2 patients who underwent duplex scanning, there were 18 (13.3%) noted to have DVT compared to 72 of the 711 patients (10.1%) who were either SARS-CoV-2 negative or untested. The odds ratio for DVT in COVID-19 was 1.35 (95% CI 0.78 – 2.34, p=0.289). Baseline characteristics for COVID-19 patients with and without DVT were overall similar. COVID-19 patients with DVT had an elevated median first D-dimer (18.88 mcg/mL [IQR 7.79, 20.00] versus 2.55 mcg/mL [IQR 1.45, 6.28], p=0.002, reference range <0.5 mcg/mL), average in-hospital D-dimer (median 11.93 mcg/mL [IQR 8.25, 16.97] versus 3.54 mcg/mL [IQR 2.05, 8.53] , p<0.001) and median fibrinogen level (501.0 (IQR 440.0, 629.0) versus 654.5 (IQR 535.8, 780.0), p= 0.002, reference range 187-502 mg/dL). There was a trend to significance for COVID-19 patients with DVT compared to without DVT in median D-dimer levels at the time of the duplex (13.61 mcg/mL [IQR 4.04, 19.97] versus 3.58 mcg/mL [IQR 2.51, 9.62], p=0.055) and median ferritin levels (1,679.0 ng/mL [IQR 1,168.0, 2,577.0] versus 1,103.0 ng/mL [IQR 703.5, 2,076.5], p=0.055, reference 25-270 ng/mL). Twelve of the 18 patients with COVID who developed DVT did so despite chemical thromboprophylaxis, and two developed DVT in spite of therapeutic anticoagulation CONCLUSIONS: We found only a modestly increased risk of DVT in patients with COVID-19, likely underestimated due to limitations in duplex testing early in the epidemic. Elevated D-dimer and a less elevated fibrinogen are associated with DVT in patients with COVID-19 who appear to form thrombus despite conventional chemical thromboprophylaxis. Additionally, an increasing D-dimer over time may be a reflection of the development of DVT in patients with COVID-19.


title: Knowledge, Perceptions, and Attitude of Egyptians Towards the Novel Coronavirus Disease (COVID-19)

text: Coronavirus disease 2019 (COVID-19) has been recognized as a pandemic by the World Health Organization. Global efforts have been exerted to prevent the spreading of the disease through political decisions together with personal behaviors, which depend on awareness of the public. The goal of this study is to assess the knowledge, perceptions and attitude of the Egyptian public towards the COVID-19 disease. We conducted a cross-sectional survey about these points, which was distributed among adult Egyptians. Five hundred and fifty nine persons completed the survey. The mean knowledge score was 16.39 out of 23, gained mainly though social media (66.9%), and the internet (58.3%). Knowledge was significantly lower among older, less educated, lower income participants, and rural residents. Most participants (86.9%) were concerned about the risk of infection. While 37.6% thought that their salary will be continued if they become isolated, 68.5% believed that it should be continued during this period. About 73.0% were looking forward to get the vaccine when available. In general, participants had a good knowledge about the disease and a positive attitude towards protective measures. This knowledge is gained mainly through novel media channels, which have pros and cons. Although the government has taken major steps to educate the public and limit the spread of the disease, more effort is needed to educate and support the lower economic strata. If a vaccine or a treatment is approved, we recommend a government control over its use to preserve the rights of the vulnerable and needy groups. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10900-020-00827-7) contains supplementary material, which is available to authorized users.


title: Identification and complete genome analysis of a novel bovine picornavirus in Japan

text: Abstract We identified novel viruses in feces from cattle with diarrhea collected in 2009 in Hokkaido Prefecture, Japan, by using a metagenomics approach and determined the (near) complete sequences of the virus. Sequence analyses revealed that they had a standard picornavirus genome organization, i.e. 5′ untranslated region (UTR) - L- P1 (VP4- VP3- VP2- VP1) - P2 (2A- 2B- 2C) - P3 (3A- 3B- 3C-3D) - 3′UTR- poly(A). They are closely related to other unclassified Chinese picornaviruses; bat picornaviruses group 1–3, feline picornavirus, and canine picornavirus, sharing 45.4–51.4% (P1), 38.0–44.9% (P2), and 49.6–53.3% (P3) amino acid identities, respectively. The phylogenetic analyses and detailed genome characterization showed that they, together with the unclassified Chinese picornaviruses, grouped as a cluster for the P1, 2C, 3CD and VP1 coding regions. These viruses had conserved features (e.g. predicted protein cleavage sites, presence of a leader protein, 2A, 2C, 3C, and 3D functional domains), suggesting they have a common ancestor. Reverse-transcription-PCR assays, using specific primers designed from the 5′UTR sequence of these viruses, showed that 23.0% (20/87) of fecal samples from cattle with diarrhea were positive, indicating the prevalence of these picornavirus in the Japanese cattle population in Hokkaido Prefecture. However, further studies are needed to investigate the pathogenic potential and etiological role of these viruses in cattle.


title: Chapter 5 Laboratory diagnosis of viral infection

text: Abstract Since the last Handbook of Clinical Neurology volume on this topic, viral diagnosis has made tremendous strides, moving from the margin to the mainstream of clinical care. For many years, conventional virus isolation was the mainstay of viral diagnosis since it was sensitive and “open-minded.” However, growth in conventional cell culture entails an inherent delay that limits its clinical impact. Although rapid culture and viral antigen methods detect fewer pathogens and are less sensitive than conventional culture, both require less expertise and have greatly reduced time to result. Polymerase chain reaction has ushered in a new era in virology, especially in the diagnosis of neurologic diseases. Molecular amplification methods are rapid, highly sensitive, can be automated, quantitative, and detect viruses not amenable to routine culture. User-friendly, walk-away tests with results in an hour, as well as multiplex tests that can detect 20 viruses in a single reaction, are now a reality. As the variety of test methods and commercial products proliferate, the challenges for clinicians and laboratories are selecting which tests to utilize in which clinical scenarios, and understanding how to interpret the results. The advantages and limitations of each method are discussed in this chapter, with special attention to neurologic disease.


title: Bovine coronavirus in naturally and experimentally exposed calves; viral shedding and the potential for transmission

text: BACKGROUND: Bovine coronavirus (BCoV) is a widely distributed pathogen, causing disease and economic losses in the cattle industry worldwide. Prevention of virus spread is impeded by a lack of basic knowledge concerning viral shedding and transmission potential in individual animals. The aims of the study were to investigate the duration and quantity of BCoV shedding in feces and nasal secretions related to clinical signs, the presence of virus in blood and tissues and to test the hypothesis that seropositive calves are not infectious to naïve in-contact calves three weeks after BCoV infection. METHODS: A live animal experiment was conducted, with direct contact between animal groups for 24 h as challenge procedure. Four naïve calves were commingled with a group of six naturally infected calves and sequentially euthanized. Two naïve sentinel calves were commingled with the experimentally exposed group three weeks after exposure. Nasal swabs, feces, blood and tissue samples were analyzed for viral RNA by RT-qPCR, and virus isolation was performed on nasal swabs. Serum was analyzed for BCoV antibodies. RESULTS: The calves showed mild general signs, and the most prominent signs were from the respiratory system. The overall clinical score corresponded well with the shedding of viral RNA the first three weeks after challenge. General depression and cough were the signs that correlated best with shedding of BCoV RNA, while peak respiratory rate and peak rectal temperature appeared more than a week later than the peak shedding. Nasal shedding preceded fecal shedding, and the calves had detectable amounts of viral RNA intermittently in feces through day 35 and in nasal secretions through day 28, however virus isolation was unsuccessful from day six and day 18 from the two calves investigated. Viral RNA was not detected in blood, but was found in lymphatic tissue through day 42 after challenge. Although the calves were shedding BCoV RNA 21 days after infection the sentinel animals were not infected. CONCLUSIONS: Prolonged shedding of BCoV RNA can occur, but detection of viral RNA does not necessarily indicate a transmission potential. The study provides valuable information with regard to producing scientifically based biosecurity advices. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-016-0555-x) contains supplementary material, which is available to authorized users.


title: Antibody-mediated Neutralization of Ebola Virus Can Occur by Two Distinct Mechanisms

text: Human Ebola virus (EBOV) causes severe hemorrhagic fever disease with high mortality and there is no vaccine or treatment. Antibodies in survivors occur early, are sustained, and can delay infection when transferred into nonhuman primates. Monoclonal antibodies (mAbs) from survivors exhibit potent neutralizing activity in vitro and are protective in rodents. To better understand targets and mechanisms of neutralization, we investigated a panel of mAbs shown previously to react with the envelope glycoprotein (GP). While one non-neutralizing mAb recognized a GP epitope in the non-essential mucin-like domain, the rest were specific for GP1, were neutralizing, and could be further distinguished by reactivity with secreted GP. We show that survivor antibodies, human KZ52 and monkey JP3K11, were specific for conformation-dependent epitopes comprising residues in GP1 and GP2 and that neutralization occurred by two distinct mechanisms; KZ52 inhibited cathepsin cleavage of GP whereas JP3K11 recognized the cleaved, fusion-active form of GP.


title: Sequence motifs and prokaryotic expression of the reptilian paramyxovirus fusion protein

text: Fourteen reptilian paramyxovirus isolates were chosen to represent the known extent of genetic diversity among this novel group of viruses. Selected regions of the fusion (F) gene were sequenced, analyzed and compared. The F gene of all isolates contained conserved motifs homologous to those described for other members of the family Paramyxoviridae including: signal peptide, transmembrane domain, furin cleavage site, fusion peptide, N-linked glycosylation sites, and two heptad repeats, the second of which (HRB-LZ) had the characteristics of a leucine zipper. Selected regions of the fusion gene of isolate Gono-GER85 were inserted into a prokaryotic expression system to generate three recombinant protein fragments of various sizes. The longest recombinant protein was cleaved by furin into two fragments of predicted length. Western blot analysis with virus-neutralizing rabbit-antiserum against this isolate demonstrated that only the longest construct reacted with the antiserum. This construct was unique in containing 30 additional C-terminal amino acids that included most of the HRB-LZ. These results indicate that the F genes of reptilian paramyxoviruses contain highly conserved motifs typical of other members of the family and suggest that the HRB-LZ domain of the reptilian paramyxovirus F protein contains a linear antigenic epitope.


title: Intra-abdominal hypertension as a trigger of “gut failure” in SARS-CoV-2 infection: effect of open abdomen (OA) and negative pressure therapy (NPT) on respiratory and gastrointestinal (GI) function

text: COVID-19 gastrointestinal manifestations could be attributed to SARS-CoV-2-induced small vessel thrombosis. OA with NPT treatment may have a role in optimization of bowel microcirculation and in the reduction of the endothelial and the systemic cytokine-related damage, improving also respiratory function.


title: Is schizophrenia research relevant during the COVID-19 pandemic?

text: Abstract Schizophrenia researchers may ask themselves—or be asked by others—whether their research is relevant in the face of the COVID-19 pandemic. This commentary argues that schizophrenia research is more relevant than ever during this public health crisis, because of the likelihood that the COVID-19 pandemic may lead to increased incidence of psychotic disorders.


Epizootic Catarrhal Enteritis