title: Chapter Two Structures of enveloped virions determined by cryogenic electron microscopy and tomography

text: Abstract Enveloped viruses enclose their genomes inside a lipid bilayer which is decorated by membrane proteins that mediate virus entry. These viruses display a wide range of sizes, morphologies and symmetries. Spherical viruses are often isometric and their envelope proteins follow icosahedral symmetry. Filamentous and pleomorphic viruses lack such global symmetry but their surface proteins may display locally ordered assemblies. Determining the structures of enveloped viruses, including the envelope proteins and their protein-protein interactions on the viral surface, is of paramount importance. These structures can reveal how the virions are assembled and released by budding from the infected host cell, how the progeny virions infect new cells by membrane fusion, and how antibodies bind surface epitopes to block infection. In this chapter, we discuss the uses of cryogenic electron microscopy (cryo-EM) in elucidating structures of enveloped virions. Starting from a detailed outline of data collection and processing strategies, we highlight how cryo-EM has been successfully utilized to provide unique insights into enveloped virus entry, assembly, and neutralization.


title: Immunoproteomic analysis of bacterial proteins of Actinobacillus pleuropneumoniae serotype 1

text: BACKGROUND: Actinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide. Identification and characterization of novel antigenic APP vaccine candidates are underway. In the present study, we use an immunoproteomic approach to identify APP protein antigens that may elicit an immune response in serotype 1 naturally infected swine and serotype 1 virulent strain S259-immunized rabbits. RESULTS: Proteins from total cell lysates of serotype 1 APP were separated by two-dimensional electrophoresis (2DE). Western blot analysis revealed 21 immunoreactive protein spots separated in the pH 4-7 range and 4 spots in the pH 7-11 range with the convalescent sera from swine; we found 5 immunoreactive protein spots that separated in the pH 4-7 range and 2 in the pH 7-11 range with hyperimmune sera from S259-immunized rabbits. The proteins included the known antigens ApxIIA, protective surface antigen D15, outer membrane proteins P5, subunit NqrA. The remaining antigens are being reported as immunoreactive proteins in APP for the first time, to our knowledge. CONCLUSIONS: We identified a total of 42 immunoreactive proteins of the APP serotype 1 virulent strain S259 which represented 32 different proteins, including some novel immunoreactive factors which could be researched as vaccine candidates.


title: Proteomic analysis of purified Newcastle disease virus particles

text: BACKGROUND: Newcastle disease virus (NDV) is an enveloped RNA virus, bearing severe economic losses to the poultry industry worldwide. Previous virion proteomic studies have shown that enveloped viruses carry multiple host cellular proteins both internally and externally during their life cycle. To address whether it also occurred during NDV infection, we performed a comprehensive proteomic analysis of highly purified NDV La Sota strain particles. RESULTS: In addition to five viral structural proteins, we detected thirty cellular proteins associated with purified NDV La Sota particles. The identified cellular proteins comprised several functional categories, including cytoskeleton proteins, annexins, molecular chaperones, chromatin modifying proteins, enzymes-binding proteins, calcium-binding proteins and signal transduction-associated proteins. Among these, three host proteins have not been previously reported in virions of other virus families, including two signal transduction-associated proteins (syntenin and Ras small GTPase) and one tumor-associated protein (tumor protein D52). The presence of five selected cellular proteins (i.e., β-actin, tubulin, annexin A2, heat shock protein Hsp90 and ezrin) associated with the purified NDV particles was validated by Western blot or immunogold labeling assays. CONCLUSIONS: The current study presented the first standard proteomic profile of NDV. The results demonstrated the incorporation of cellular proteins in NDV particles, which provides valuable information for elucidating viral infection and pathogenesis.


title: Isolation rooms for highly infectious diseases: an inventory of capabilities in European countries

text: Summary Isolation of patients with highly infectious diseases (HIDs) in hospital rooms with adequate technical facilities is essential to reduce the risk of spreading disease. The European Network for Infectious Diseases (EUNID), a project co-funded by European Commission and involving 16 European Union member states, performed an inventory of high level isolation rooms (HIRs, hospital rooms with negative pressure and anteroom). In participating countries, HIRs are available in at least 211 hospitals, with at least 1789 hospital beds. The adequacy of this number is not known and will depend on prevailing circumstances. Sporadic HID cases can be managed in the available HIRs. HIRs could also have a role in the initial phases of an influenza pandemic. However, large outbreaks due to natural or to bioterrorist events will need management strategies involving healthcare facilities other than HIRs.


title: High-resolution CT features of 17 cases of Corona Virus Disease 2019 in Sichuan province, China

text: Bilateral ground-glass opacities and a combination of consolidation and ground-glass opacities mainly in the subpleural lung regions is a noteworthy high-resolution CT feature of Corona Virus Disease 2019 which may help in the early diagnosis of the disease.


title: How villagers in central Sierra Leone understand infection risks under threat of Covid-19

text: BACKGROUND: Concern has been expressed over how well Africa is prepared to cope with the pandemic of Covid-19. Will rural populations with low levels of education know how to apply community-based infection control? We undertook fieldwork in two villages in central Sierra Leone to gain insight into how rural people faced with Covid-19 assess epidemic infection risks. METHODS: Two communities were selected based on prior contrasted exposure to Ebola Virus Disease–one with substantial number of cases and the other having resisted infection through strong community sequestration measures. We assessed understanding of infection risks via an experimental game. This asked players to express a preference for one of two diseases, one resembling Ebola with lower risk of infection and the other resembling Covid-19 with lower risk of death. Players were not told the identity of the diseases. RESULTS: In total 107 adult villagers played the game (58% women). Half (52%) preferred the disease model with lower risk of infection, 29% preferred the model with lower risk of death, while 21% saw the combined risk of infection and death as being equivalent. Differences in reactions between the two locations were small despite different experiences of Ebola. Asked to explain their choices 48% of players cited information on infection risks modelled by the game and 31% stated that their choices reflected awareness of the need for personal action and respect for local regulations. We concluded that villagers thoughtfully assess disease risks and that some are good intuitive statisticians. CONCLUSIONS: Results suggest rural people in Sierra Leone retain the lessons of experience from the Ebola outbreak of 2014–15 and will be able to apply these lessons to a new infectious disease for which have no prior practical experience. Our expectation is that rural populations will understand Covid-19 control measures, thus reducing need for draconian enforcement.


title: The Anti-Porcine Parvovirus Activity of Nanometer Propolis Flavone and Propolis Flavone In Vitro and In Vivo

text: Objectives. The present study was conducted to evaluate the activity of nanometer propolis flavone (NPF) on inhibiting porcine parvovirus (PPV) in vitro and in vivo. Methods. In vitro, the effect of NPF on cellular infectivity of PPV was carried out before and after adding drug and simultaneous adding and PPV after being mixed. In vivo, the anti-PPV effect of NPF in guinea pigs was performed. Results. The results showed that NPF could significantly inhibit PPV infecting porcine kidney- (PK-) 15 cells compared with propolis flavone (PF), and the activity of NPF was the best in preadding drug pattern. NPF at high and medium doses was able to observably restrain PPV copying in lung, gonad, blood, and spleen, decrease the impact of PPV on weight of guinea pigs, and improve hemagglutination inhibition (HI) of PPV in serum. In addition, it could also increase the contents of IL-2 and IL-6 in serum after PPV challenge. Conclusion. These results indicated that NPF could significantly improve the anti-PPV activity of PF, and its high concentration possessed the best efficacy. Therefore, NPF would be expected to be exploited into a new-style antiviral drug.


title: Stability Analysis and Numerical Simulation of SEIR Model for pandemic COVID-19 spread in Indonesia

text: The Aim of this research is construct the SEIR model for COVID-19, Stability Analysis and numerical simulation of the SEIR model on the spread of COVID-19. The method used to construct the model is the SEIR model by considering vaccination and isolation factors as model parameters, the analysis of the model uses the generation matrix method to obtain the basic reproduction numbers and the global stability of the COVID-19 distribution model. Numerical simulation models use secondary data on the number of COVID-19 cases in Indonesia. The results obtained are the SEIR model for COVID-19; model analysis yields global stability from the spread of COVID-19; The results of the analysis also provide information if no vaccine, Indonesia is endemic COVID-19. Then the simulation results provide a prediction picture of the number of COVID-19 in Indonesia in the following days, the simulation results also show that the vaccine can accelerate COVID-19 healing and maximum isolation can slow the spread of COVID-19. The results obtained can be used as a reference for early prevention of the spread of COVID-19 in Indonesia


Covid‐19


title: Chapter 45 Gender Differences in Emerging Infectious Diseases

text: Publisher Summary This chapter focuses on gender differences in emerging infectious diseases. An urgent worldwide threat is posed by the introduction and spread of novel infectious diseases. The reasons for emerging infectious diseases are numerous and complex. Among the most significant explanations for these emerging diseases are changes in environment and ecology caused by natural phenomena such as droughts, hurricanes, and floods; and human-made phenomena such as agricultural development, urbanization, and denuding of forests. Worldwide conflict, including wars, ethnic cleansing, and genocide, have led to displacement of large populations into overcrowded settlements where safe water is not available and sanitation is poor. For example, unsanitary conditions led to a huge increase in the rat population in post-war Kosovo, resulting in a tularemia outbreak with 327 confirmed cases in 8 months. Regional conflict leads to breakdown in infection control, inadequate surveillance, impeded access to populations, and spread of infectious diseases through movement of refugees and aid workers. Increased precipitation, a result of climate change, leads to more agricultural run-off, allowing pathogens to enter drinking water systems. In developing countries where poverty and inadequate infrastructure are the norm, public health monitoring systems must be supported and improved so that new or more severe risks to health can be identified and curtailed. As new infectious diseases are recognized, critical issues arise regarding pregnant women and their unborn children. Physiologic changes during pregnancy and gestational age both alter decision-making regarding vaccinations and medications.


title: Online Gambling in the Midst of COVID-19: A Nexus of Mental Health Concerns, Substance Use and Financial Stress

text: This study examined the emerging impact of COVID-19 on gambling during the first 6 weeks of emergency measures in Ontario, Canada. A cross-sectional online survey of 2005 gamblers, including a sub-sample of 1081 online gamblers (age 18 years and older), was administered to assess risky gambling behaviours and motivations, financial impacts from COVID-19, the influence of COVID-19 on online gambling, mental health concerns and substance use. A series of odds ratio comparisons and measures of association were carried out. Results show significant likelihood of online gambling among those classified as high-risk gamblers (according to the Problem Gambling Severity Index) and those with past experience of online gambling, though migration from land-based gambling was apparent. Among high-risk online gamblers, the most predictive risk factors included moderate and severe anxiety and depression, reduced work hours, being influenced to gamble due to COVID-19, gambling under the influence of cannabis or alcohol and risky gambling motives tied to mental health concerns, including gambling because it helps with nervousness and depression, chasing gambling losses and seeking to earn income. This study has confirmed many of the risk associations presented in emerging COVID-19-related studies and past research on global economic crisis relating to gambling risk, mental health concerns and substance use. However, unlike many past studies, the present paper takes note of all of these elements holistically and provides incremental clarity on online gambling risk factors during the pandemic, specifically.


Index to volume 6


title: COVID-19 (SARS-Cov-2) and the heart – An ominous association

text: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Cardiovascular manifestations of COVID-19 are diverse and complex and include acute coronary syndrome, myocarditis masquerading as ST-segment elevation myocardial infarction, pericarditis and pericardial effusion. We present 2 cases of COVID-19 infection with myocardial involvement with distinct mechanistic pathways and outcomes. Important decision strategies such as the timing of cardiac catheterization (when indicated) and requirement of early hemodynamic support in critically ill patients are discussed.


title: Factors associated with chronic obstructive pulmonary disease exacerbation, based on big data analysis

text: Preventing exacerbation in chronic obstructive pulmonary disease (COPD) patients is crucial, but requires identification of the exacerbating factors. To date, no integrated analysis of patient-derived and external factors has been reported. To identify factors associated with COPD exacerbation, we collected data, including smoking status, lung function, and COPD assessment test scores, from 594 COPD patients in the Korean COPD subgroup study (KOCOSS), and merged these data with patients’ Korean Health Insurance Review and Assessment Service data for 2007–2012. We also collected primary weather variables, including levels of particulate matter <10 microns in diameter, daily minimum ambient temperature, as well as respiratory virus activities, and the logs of web queries on COPD-related issues. We then assessed the associations between these patient-derived and external factors and COPD exacerbations. Univariate analysis showed that patient factors, air pollution, various types of viruses, temperature, and the number of COPD-related web queries were associated with COPD exacerbation. Multivariate analysis revealed that the number of exacerbations in the preceding year, female sex, COPD grade, and influenza virus detection rate, and lowest temperature showed significant association with exacerbation. Our findings may help COPD patients predict when exacerbations are likely, and provide intervention as early as possible.


title: The structural and accessory proteins M, ORF 4a, ORF 4b, and ORF 5 of Middle East respiratory syndrome coronavirus (MERS-CoV) are potent interferon antagonists

text: The newly emerged Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus with pathogenic mechanisms that may be driven by innate immune pathways. The goal of this study is to characterize the expression of the structural (S, E, M, N) and accessory (ORF 3, ORF 4a, ORF 4b, ORF 5) proteins of MERS-CoV and to determine whether any of these proteins acts as an interferon antagonist. Individual structural and accessory protein-coding plasmids with an N-terminal HA tag were constructed and transiently transfected into cells, and their native expression and subcellular localization were assessed using Wes tern blotting and indirect immunofluorescence. While ORF 4b demonstrated majorly nuclear localization, all of the other proteins demonstrated cytoplasmic localization. In addition, for the first time, our experiments revealed that the M, ORF 4a, ORF 4b, and ORF 5 proteins are potent interferon antagonists. Further examination revealed that the ORF 4a protein of MERS-CoV has the most potential to counteract the antiviral effects of IFN via the inhibition of both the interferon production (IFN-β promoter activity, IRF-3/7 and NF-κB activation) and ISRE promoter element signaling pathways. Together, our results provide new insights into the function and pathogenic role of the structural and accessory proteins of MERS-CoV.


title: Proteome Profile of Swine Testicular Cells Infected with Porcine Transmissible Gastroenteritis Coronavirus

text: The interactions occurring between a virus and a host cell during a viral infection are complex. The purpose of this paper was to analyze altered cellular protein levels in porcine transmissible gastroenteritis coronavirus (TGEV)-infected swine testicular (ST) cells in order to determine potential virus-host interactions. A proteomic approach using isobaric tags for relative and absolute quantitation (iTRAQ)-coupled two-dimensional liquid chromatography-tandem mass spectrometry identification was conducted on the TGEV-infected ST cells. The results showed that the 4-plex iTRAQ-based quantitative approach identified 4,112 proteins, 146 of which showed significant changes in expression 48 h after infection. At 64 h post infection, 219 of these proteins showed significant change, further indicating that a larger number of proteomic changes appear to occur during the later stages of infection. Gene ontology analysis of the altered proteins showed enrichment in multiple biological processes, including cell adhesion, response to stress, generation of precursor metabolites and energy, cell motility, protein complex assembly, growth, developmental maturation, immune system process, extracellular matrix organization, locomotion, cell-cell signaling, neurological system process, and cell junction organization. Changes in the expression levels of transforming growth factor beta 1 (TGF-β1), caspase-8, and heat shock protein 90 alpha (HSP90α) were also verified by western blot analysis. To our knowledge, this study is the first time the response profile of ST host cells following TGEV infection has been analyzed using iTRAQ technology, and our description of the late proteomic changes that are occurring after the time of vigorous viral production are novel. Therefore, this study provides a solid foundation for further investigation, and will likely help us to better understand the mechanisms of TGEV infection and pathogenesis.


title: Superabsorbent Fibers for Comfortable Disposable Medical Protective Clothing

text: Disposable medical protective clothing for 2019-nCoV mainly consists of stacked layers with nanopore films, polymer coated nonwoven fabrics and melt-blown nonwoven fabrics against anti-microbial and anti-liquid penetration. However, such structures lack moisture permeability and breathability leading to an uncomfortable, stuffy wearing experience. Here, we propose a novel medical protective clothing material with a superabsorbent layer to enhance moisture absorption. Poly(acrylic acid-co-acrylamide)/polyvinyl alcohol superabsorbent fibers (PAAAM/PVA fibers) were prepared via wet spinning. And the superabsorbent composite layer was stacked from PAAAM/PVA fibers, bamboo pulp fibers (BPF) and ethylene-propylene side by side fibers (ESF). The novel disposable medical protective composite fabric was obtained through gluing the superabsorbent layer to the inner surface of strong antistatic polypropylene nonwoven fabric. The resultant composite fabric possesses excellent absorption and retention capacity for sweat, up to 12.3 g/g and 63.8%, and a maximum hygroscopic rate of 1.04 g/h, higher than that of the conventional material (only 0.53 g/h). The moisture permeability of the novel material reached 12,638.5 g/(m(2) d), which was 307.6% of the conventional material. The novel material can effectively reduce the humidity inside the protective clothing and significantly improve the comfort of medical staff.


title: Demonstration of a difference in expression of maximal lactase and sucrase activity along the villus in the adult rat jejunum

text: Abstract Lactase and sucrase are two disaccharidases that differ not only in their substrate specificity and developmental patterns, but also in their resistance to mucosal insult. In this experiment, we tested the hypothesis that there might be a dichotomy in expression of enzyme activity along the jejunal villuscrypt unit. Sectioning of the villus-crypt unit in a cryostat enabled direct comparison of the distribution of lactase and sucrase enzyme activities in the adult rat. There is a stepwise increase in mean lactase/sucrase ratio going from crypt to villus. The data indicate that unlike sucrase activity, which is expressed maximally in enterocytes along the entire villus, maximal lactase activity is not attained until midvillus. The delay in expression of maximal lactase activity might help to explain the vulnerability of this enzyme to acute mucosal insult such as occurs in viral gastroenteritis.


title: IFIT3 and IFIT2/3 promote IFIT1-mediated translation inhibition by enhancing binding to non-self RNA

text: Interferon-induced proteins with tetratricopeptide repeats (IFITs) are highly expressed during the cell-intrinsic immune response to viral infection. IFIT1 inhibits translation by binding directly to the 5′ end of foreign RNAs, particularly those with non-self cap structures, precluding the recruitment of the cap-binding eukaryotic translation initiation factor 4F and subsequent 40S recruitment. Interaction of different IFIT family members is well described, but little is known of the molecular basis of IFIT association or its impact on function. Here, we reconstituted different complexes of IFIT1, IFIT2 and IFIT3 in vitro, which enabled us to reveal critical aspects of IFIT complex assembly. IFIT1 interacts rapidly and strongly with IFIT3 forming a stable heterotetramer. IFIT2 and IFIT3 homodimers dissociate to form a more stable heterodimer that associates with IFIT1, forming an IFIT1:IFIT2:IFIT3 trimer. Site-directed mutagenesis revealed a C-terminal ‘YxxxL’ motif in IFIT1 that mediates its association with IFIT3. Using various reporter mRNAs, we demonstrate for the first time that IFIT3 stabilises IFIT1 binding to cap0-mRNA and enhances its translation inhibition activity. Disrupting the binding interface between IFIT1 and IFIT3 abrogated this enhancement. This work reveals molecular aspects of IFIT assembly and provides an important ‘missing link’ between IFIT interaction and function.


Treating Influenza in the Emergency Department


title: Human herpesviruses respiratory infections in patients with acute respiratory distress (ARDS)

text: Acute respiratory distress syndrome (ARDS) is today a leading cause of hospitalization in intensive care unit (ICU). ARDS and pneumonia are closely related to critically ill patients; however, the etiologic agent is not always identified. The presence of human herpes simplex virus 1, human cytomegalovirus and Epstein–Barr virus in respiratory samples of critically ill patients is increasingly reported even without canonical immunosuppression. The main aim of this study was to better understand the significance of herpesviruses finding in lower respiratory tract of ARDS patients hospitalized in ICU. The presence of this group of herpesviruses, in addition to the research of influenza viruses and other common respiratory viruses, was investigated in respiratory samples from 54 patients hospitalized in ICU, without a known microbiological causative agent. Moreover, the immunophenotype of each patient was analyzed. Herpesviruses DNA presence in the lower respiratory tract seemed not attributable to an impaired immunophenotype, whereas a significant correlation was observed between herpesviruses positivity and influenza virus infection. A higher ICU mortality was significantly related to the presence of herpesvirus infection in the lower respiratory tract as well as to impaired immunophenotype, as patients with poor outcome showed severe lymphopenia, affecting in particular T (CD3+) cells, since the first days of ICU hospitalization. In conclusion, these results indicate that herpesviruses lower respiratory tract infection, which occurs more frequently following influenza virus infection, can be a negative prognostic marker. An independent risk factor for ICU patients with ARDS is an impaired immunophenotype.


title: Middle East Respiratory Syndrome Coronavirus Infection Dynamics and Antibody Responses among Clinically Diverse Patients, Saudi Arabia

text: Middle East respiratory syndrome coronavirus (MERS-CoV) shedding and antibody responses are not fully understood, particularly in relation to underlying medical conditions, clinical manifestations, and mortality. We enrolled MERS-CoV–positive patients at a hospital in Saudi Arabia and periodically collected specimens from multiple sites for real-time reverse transcription PCR and serologic testing. We conducted interviews and chart abstractions to collect clinical, epidemiologic, and laboratory information. We found that diabetes mellitus among survivors was associated with prolonged MERS-CoV RNA detection in the respiratory tract. Among case-patients who died, development of robust neutralizing serum antibody responses during the second and third week of illness was not sufficient for patient recovery or virus clearance. Fever and cough among mildly ill patients typically aligned with RNA detection in the upper respiratory tract; RNA levels peaked during the first week of illness. These findings should be considered in the development of infection control policies, vaccines, and antibody therapeutics.


title: Clinical characteristics of 80 hospitalized frontline medical workers infected with COVID-19 in Wuhan, China

text: Summary More than 1,000 medical workers have been infected with COVID-19 in China. The 80 hospitalized patients included 57 SARS-CoV-2 confirmed and 23 clinically diagnosed. The median age was 39 years, 49 (61.25%) were women, and one patient died. The most common symptoms at onset were fever (65, 81.25%), cough (47, 58.75%), fatigue (28, 35%), myalgia (19, 23.75%), expectoration (19, 23.75%), and diarrhea (15, 18.75%). Patients of frontline medical workers at a single-center hospital showed some unique clinical and laboratory findings compared with other patients in Wuhan and outside of Wuhan. This study provides our experience for other frontline medical workers.


Impact of SARS on development of childhood acute lymphoblastic leukaemia


title: HIV care in times of the COVID-19 crisis – where are we now in Central and Eastern Europe?

text: Abstract Introduction The SARS-CoV-2 pandemic hit disproportionately European region. Many HIV clinics share staff and logistics with infectious disease facilities, which are now on the frontline of COVID-19. Therefore, we investigated the impact of current pandemic situation on HIV care and continuity of antiretroviral treatment (ART) supplies in CEE countries. Methods Euroguidelines in Central and Eastern Europe (ECEE) Network Group was established in February 2016 to review standards of care for HIV in the region. The group consists of professionals actively involved in HIV care. On 19th March we decided to review the status of HIV care sustainability in the face of emerging SARS-CoV-2 pandemic in Europe. For this purpose, we constructed an on-line survey consisting of 23 questions. Respondents were recruited from the ECEE members in 22 countries, based on their involvement in HIV care and contacted through e-mails. Results In total 19 countries responded: Albania, Armenia, Belarus, Bosnia and Herzegovina, Bulgaria, Croatia, Czech Republic, Estonia, Georgia, Greece, Hungary, Lithuania, Macedonia, Poland, Republic of Moldova, Russia, Serbia, Turkey, Ukraine. Most of the respondents were infectious disease physicians directly involved in HIV care (17/19). No country reported HIV clinic closure. HIV clinics were operating normally in only six countries (31.6%). In 11 (57.9%) countries physicians were sharing HIV care and COVID-19 care duties. None of the countries expect shortage of ART in the next two weeks; however five physicians expressed uncertainty about the next two months. At the time of providing responses ten (52.6%) countries had HIV-positive persons under quarantine. Conclusions The shortage of resources is evident and the impact on HIV care inevitable. We need to prepare for operating under minimal medical resources with the aim to secure retention on ART. Non-governmental organizations should re-validate their earlier objectives and second medical efforts to ascertain the continuity of ART deliverance.


What Happens Next: Radiation Oncology After COVID?


title: Isolation and characterization of a variant subgroup GII-a porcine epidemic diarrhea virus strain in China

text: Abstract Background Highly virulent variants of porcine epidemic diarrhea virus (PEDV) have been closely associated with recent outbreaks of porcine epidemic diarrhea (PED) in China, which have resulted in severe economic losses to the pork industry. Methods In the current study, the variant PEDV strain HM2017 was isolated and purified and a viral growth curve was constructed according to the median tissue culture infective dose (TCID50). HM2017 were amplify with RT-PCR and analyzed by phylogeny analysis. Animal pathogenicity experiment was carried to evaluate the HM2017 clinical assessment. Results Genome-based phylogenetic analysis revealed that PEDV strain HM2017 was clustered into the variant subgroup GII-a that is currently circulating in pig populations in China. The highest median tissue culture infectious dose of strain HM2017 after 15 passages in Vero cells was 1.33 × 107 viral particles/mL. Strain HM2017 was highly virulent to suckling piglets, which exhibited clinical symptoms at 12 h post-infection (hpi) (i.e., weight loss at 12–84 hpi, increased body temperatures at 24–48 hpi, high viral loads in the jejunum and ileum, and 100% mortality by 84 hpi). Conclusion The present study reports a variant subgroup GII-a PEDV HM2017 strain in China and characterize its pathogenicity. PEDV strain HM2017 of subgroup GII-a presents a promising vaccine candidate for the control of PED outbreaks in China.


title: Vaccines in Veterinary Medicine: A Brief Review of History and Technology

text: The use of vaccines in veterinary medicine has progressed from an experimental adventure to a routine and relatively safe practice. The common and aggressive use of efficacious vaccines has been responsible for the control and eradication of several diseases. Despite progress in research technologies, diagnostic capabilities, and manufacturing methods, there remain many infectious diseases for which no effective vaccines exist. Global availability, field compliance, effectiveness, and safety are also significant concerns. This review addresses the history, current practices, and potential future improvements of vaccine use in veterinary medicine.


title: Phylogenetic inference from binary sequences reduced by primary DNA sequences

text: Given a bi-classification of nucleotides, we can obtain a reduced binary sequence of a primary DNA sequence. This binary sequence will undoubtedly retain some biological information and lose the rest. Here we want to know what kind of and how much biological information an individual binary sequence carries. Three classifications of nucleotides are explored in the present article. Phylogenetic trees are built from these binary sequences by the Neighbor-Joining (NJ) method, with evolutionary distance evaluated on the basis of a symbolic sequence complexity. We find that, for all data sets studied, binary sequences reduced by the purine/pyrimidine classification give reliable phylogeny (almost the same as that from the primary sequences), while the other two result in discrepancies at different levels. Some possible reasons and a simple model of sequence evolutionary are introduced to interpret this phenomenon.


title: The ubiquity of asymptomatic respiratory viral infections in the tonsils and adenoids of children and their impact on airway obstruction

text: Abstract Background Airway obstruction due to enlargement of tonsils and adenoids is a common pediatric problem resulting in sleep disordered breathing. The cause for the relatively abnormal growth of tonsils and adenoids is poorly understood. Methods Non-acutely ill children undergoing tonsillectomy and adenoidectomy (T&A) for various reasons were enrolled prospectively in a study to determine the frequency of asymptomatic respiratory viral infections in each lymphoid tissue and to relate the number and types of virus to the degree of airway obstruction. Molecular techniques were used to detect 9 respiratory viruses while Brodsky scores and measurements of percentages airway obstruction were used to estimate the degree of airway compromise due to the tonsil and adenoid, respectively. Results Viruses were detected in 70.9% of tonsils and 94.7% of adenoids, p < 0.001. Adenovirus was the most common virus detected at 71.1%. Adenoids had an average of 2.4 viruses compared to 0.92 for tonsils, p < 0.001. Higher Brodsky scores were only associated with EBV in tonsils, p = 0.03, while greater percentages of airway obstruction in the adenoids were associated with adenovirus, EBV, corona virus, parainfluenza virus and rhinovirus, p ≤ 0.005. Conclusions Asymptomatic viral infections are common and directly related to the degree of airway obstruction significantly more often in adenoids than tonsils.


Public’s early response to the novel coronavirus–infected pneumonia


title: Evidence from Cochrane systematic reviews for controlling the dissemination of COVID-19 infection. A narrative review

text: BACKGROUND: COVID-19 infection has high transmissibility and several measures have been adopted for controlling its dissemination OBJECTIVE: To identify and summarize the evidence from Cochrane systematic reviews (SRs) regarding measures for controlling the dissemination of COVID-19 infection DESIGN AND SETTING: This review of Cochrane SRs was carried out in the Division of Vascular and Endovascular Surgery and in the Division of Emergency Medicine and Evidence-Based Medicine of Universidade Federal de Sao Paulo, Brazil METHODS: A comprehensive search in the Cochrane Database of Systematic Reviews retrieved all Cochrane SRs directly related to measures for controlling COVID-19 dissemination The main characteristics and results of all the SRs included were summarized and discussed RESULTS: Three Cochrane SRs were included in the qualitative synthesis These evaluated population-based and individual measures for controlling the dissemination of COVID-19 CONCLUSION: Low-certainty evidence shows that quarantine for people exposed to confirmed or suspected COVID-19 cases prevented 44% to 81% of incident cases and 31% to 63% of deaths, compared with situations of no measures Moreover, the sooner the quarantine measures were implemented, the greater the cost savings were High-confidence evidence showed that clear communication about infection control and prevention guidelines was vital for successful implementation Low-certainty evidence showed that healthcare professionals with long gowns were less exposed to contamination than were those using coveralls In addition, coveralls were more difficult to doff Further SRs on controlling the dissemination of COVID-19 infection are desirable


title: Losartan and enalapril decrease viral absorption and interleukin 1 beta production by macrophages in an experimental dengue virus infection

text: The role of angiotensin II (Ang II) in dengue virus infection remains unknown. The aim of this study was to determine the effect of losartan, an antagonist of the angiotensin II type 1 receptor (AT1 receptor), and enalapril, an inhibitor of angiotensin I-converting enzyme (ACE), on viral antigen expression and IL-1β production in peritoneal macrophages infected with dengue virus type 2. Mice treated with losartan or enalapril and untreated controls were infected intraperitoneally with the virus, and macrophages were analyzed. Infection resulted in increased IL-1β production and a high percentage of cells expressing viral antigen, and this was decreased by treatment with anti-Ang II drugs, suggesting a role for Ang II in dengue virus infection.


title: A Comprehensive Public Health Evaluation of Lockdown as a Non-pharmaceutical Intervention on COVID-19 Spread in India: National Trends Masking State Level Variations

text: INTRODUCTION: India has been under four phases of a national lockdown from March 25 to May 31 in response to the COVID-19 pandemic. Unmasking the state-wise variation in the effect of the nationwide lockdown on the progression of the pandemic could inform dynamic policy interventions towards containment and mitigation. METHODS: Using data on confirmed COVID-19 cases across 20 states that accounted for more than 99% of the cumulative case counts in India till May 31, 2020, we illustrate the masking of state-level trends and highlight the variations across states by presenting evaluative evidence on some aspects of the COVID-19 outbreak: case-fatality rates, doubling times of cases, effective reproduction numbers, and the scale of testing. RESULTS: The estimated effective reproduction number R for India was 3.36 (95% confidence interval (CI): [3.03, 3.71]) on March 24, whereas the average of estimates from May 25 - May 31 stands at 1.27 (95% CI: [1.26, 1.28]). Similarly, the estimated doubling time across India was at 3.56 days on March 24, and the past 7-day average for the same on May 31 is 14.37 days. The average daily number of tests have increased from 1,717 (March 19–25) to 131,772 (May 25–31) with an estimated testing shortfall of 4.58 million tests nationally by May 31. However, various states exhibit substantial departures from these national patterns. CONCLUSIONS: Patterns of change over lockdown periods indicate the lockdown has been effective in slowing the spread of the virus nationally. The COVID-19 outbreak in India displays large state-level variations and identifying these variations can help in both understanding the dynamics of the pandemic and formulating effective public health interventions. Our framework offers a holistic assessment of the pandemic across Indian states and union territories along with a set of interactive visualization tools that are daily updated at covind19.org.


title: Chapter 18 Molecular Basis of Pulmonary Disease

text: Publisher Summary Pulmonary pathology includes a large spectrum of both neoplastic and non-neoplastic diseases that affect the lung. Many of these are a result of the unusual relationship of the lung with the outside world. Every breath that a human takes brings the outside world into the body in the form of infectious agents, organic and inorganic particles, and noxious agents of all types. Although the lung has many defense mechanisms to protect itself from these insults, these are not infallible; therefore, lung pathology arises. Damage to the lung is particularly important given the role of the lung in the survival of the organism. Any impairment of lung function has widespread effects throughout the body, since all organs depend on the lungs for the oxygen they need. Pulmonary pathology catalogs the changes in the lung tissues and the mechanisms through which these occur. This chapter presents a review of lung pathology and the current state of knowledge about the pathogenesis of each disease. It suggests that a clear understanding of both morphology and mechanism is required for the development of new therapies and preventive measures.


title: Reduction of tumor necrosis factor production by splenocytes from v-Ha-ras oncogene-bearing mice

text: Abstract Mice containing the activated v-Ha-ras oncogene driven by the mouse mammary tumor virus promoter/enhancer produced less tumor necrosis factor (TNF) than genetically identical animals without it. Inbred Oncomice containing the v-Ha-ras oncogene and inbred FVB mice without it were grown for 6 months. Splenocytes were isolated and stimulated in vitro to produce tumor necrosis factor (TNF) and γ-interferon (IFN). TNF production by cells from Oncomice was significantly decreased compared to cells from FBV mice. There was a tendency for decrease, but no significant difference was seen on IFN release. These observations suggest that the oncogene may play a role in the immune system.


title: Factors motivating Mexico City residents to earthquake mass evacuation drills

text: Evacuation drills may constitute a key activity for preparing for an emergency due to an earthquake. The paper presents the results of an analysis of participants' motivations on the factors leading to conducting drills on 19 September every year in Mexico City; the sample size considered for the analysis was N = 2400. In particular, the following research question has been addressed: What factors predict the likelihood that respondents would report that they agree on conducting mass evacuation drills yearly? The approach has been the application of logistic regression technique to identify these factors. Of the 19 initial explanatory variables, in the final model, only seven variables and one interaction term, were significantly associated with the outcome variable; i.e.: age (Odds Ratio (OR) = 1.366; 95% Confidence Interval (CI) = 1.039–1.795); occupation (OR = 3.378; CI = 1.457–7.830); frequency of drills: one/year (OR = 2.128; CI = 1.610–2.812); knowledge vs. drills (OR = 1.394; CI = 1.172–1.658); ‘perception vulnerability city’ (OR = 1.271; CI = 1.091–1.480); warning time (OR = 1.266; CI = 1.1036–1.548); usefulness of the SASMEX (OR = 0.783; CI = 0.615-0.998); and ‘perception vulnerability city’ by occupation interaction (OR = 0.786; CI = 0.643-0.961). Further research may be needed to gain a better understanding of people's motivations on evacuation drills taking place anytime during the day or at night, and whether evacuation drills should be unannounced.


title: The CD8 T Cell Response to Respiratory Virus Infections

text: Humans are highly susceptible to infection with respiratory viruses including respiratory syncytial virus (RSV), influenza virus, human metapneumovirus, rhinovirus, coronavirus, and parainfluenza virus. While some viruses simply cause symptoms of the common cold, many respiratory viruses induce severe bronchiolitis, pneumonia, and even death following infection. Despite the immense clinical burden, the majority of the most common pulmonary viruses lack long-lasting efficacious vaccines. Nearly all current vaccination strategies are designed to elicit broadly neutralizing antibodies, which prevent severe disease following a subsequent infection. However, the mucosal antibody response to many respiratory viruses is not long-lasting and declines with age. CD8 T cells are critical for mediating clearance following many acute viral infections in the lung. In addition, memory CD8 T cells are capable of providing protection against secondary infections. Therefore, the combined induction of virus-specific CD8 T cells and antibodies may provide optimal protective immunity. Herein, we review the current literature on CD8 T cell responses induced by respiratory virus infections. Additionally, we explore how this knowledge could be utilized in the development of future vaccines against respiratory viruses, with a special emphasis on RSV vaccination.


title: Dual Mutation Events in the Haemagglutinin-Esterase and Fusion Protein from an Infectious Salmon Anaemia Virus HPR0 Genotype Promote Viral Fusion and Activation by an Ubiquitous Host Protease

text: In Infectious salmon anaemia virus (ISAV), deletions in the highly polymorphic region (HPR) in the near membrane domain of the haemagglutinin-esterase (HE) stalk, influence viral fusion. It is suspected that selected mutations in the associated Fusion (F) protein may also be important in regulating fusion activity. To better understand the underlying mechanisms involved in ISAV fusion, several mutated F proteins were generated from the Scottish Nevis and Norwegian SK779/06 HPR0. Co-transfection with constructs encoding HE and F were performed, fusion activity assessed by content mixing assay and the degree of proteolytic cleavage by western blot. Substitutions in Nevis F demonstrated that K276 was the most likely cleavage site in the protein. Furthermore, amino acid substitutions at three sites and two insertions, all slightly upstream of K276, increased fusion activity. Co-expression with HE harbouring a full-length HPR produced high fusion activities when trypsin and low pH were applied. In comparison, under normal culture conditions, groups containing a mutated HE with an HPR deletion were able to generate moderate fusion levels, while those with a full length HPR HE could not induce fusion. This suggested that HPR length may influence how the HE primes the F protein and promotes fusion activation by an ubiquitous host protease and/or facilitate subsequent post-cleavage refolding steps. Variations in fusion activity through accumulated mutations on surface glycoproteins have also been reported in other orthomyxoviruses and paramyxoviruses. This may in part contribute to the different virulence and tissue tropism reported for HPR0 and HPR deleted ISAV genotypes.


Pseudoperniosis en un adolescente durante la pandemia de COVID-19 en España


title: Current practices and barriers to the use of facemasks and respirators among hospital-based health care workers in Vietnam

text: BACKGROUND: This study aimed to examine the knowledge, attitudes, and practices towards the use of facemasks among hospital-based health care workers (HCWs) in Hanoi, Vietnam. METHODS: A qualitative study incorporating 20 focus groups was conducted between August 2010 and May 2011. HCWs from 7 hospitals in Vietnam were invited to participate. RESULTS: Issues associated with the availability of facemasks (medical and cloth masks) and respirators was the strongest theme to emerge from the discussion. Participants reported that it is not unusual for some types of facemasks to be unavailable during nonemergency periods. It was highlighted that the use of facemasks and respirators is not continuous, but rather is limited to selected situations, locations, and patients. Reuse of facemasks and respirators is also common in some settings. Finally, some participants reported believing that the reuse of facemasks, particularly cloth masks, is safe, whereas others believed that the reuse of masks put staff at risk of infection. CONCLUSIONS: In low and middle-income countries, access to appropriate levels of personal protective equipment may be restricted owing to competing demands for funding in hospital settings. It is important that issues around reuse and extended use of medical masks/respirators and decontamination of cloth masks are addressed in policy documents to minimize the risk of infection.


title: Ambulance dispatch calls attributable to influenza A and other common respiratory viruses in the Netherlands (2014‐2016)

text: BACKGROUND: Ambulance dispatches could be useful for syndromic surveillance of severe respiratory infections. We evaluated whether ambulance dispatch calls of highest urgency reflect the circulation of influenza A virus, influenza B virus, respiratory syncytial virus (RSV), rhinovirus, adenovirus, coronavirus, parainfluenzavirus and human metapneumovirus (hMPV). METHODS: We analysed calls from four ambulance call centres serving 25% of the population in the Netherlands (2014‐2016). The chief symptom and urgency level is recorded during triage; we restricted our analysis to calls with the highest urgency and identified those compatible with a respiratory syndrome. We modelled the relation between respiratory syndrome calls (RSC) and respiratory virus trends using binomial regression with identity link function. RESULTS: We included 211 739 calls, of which 15 385 (7.3%) were RSC. Proportion of RSC showed periodicity with winter peaks and smaller interseasonal increases. Overall, 15% of RSC were attributable to respiratory viruses (20% in out‐of‐office hour calls). There was large variation by age group: in <15 years, only RSV was associated and explained 11% of RSC; in 15‐64 years, only influenza A (explained 3% of RSC); and in ≥65 years adenovirus explained 9% of RSC, distributed throughout the year, and hMPV (4%) and influenza A (1%) mainly during the winter peaks. Additionally, rhinovirus was associated with total RSC. CONCLUSION: High urgency ambulance dispatches reflect the burden of different respiratory viruses and might be useful to monitor the respiratory season overall. Influenza plays a smaller role than other viruses: RSV is important in children while adenovirus and hMPV are the biggest contributors to emergency calls in the elderly.


Late‐onset fever and engraftment syndrome following autologous stem cell transplant: Impact on resource utilization


title: Genomic research delivering on promises: From rejuvenation to vaccines and pharmacogenetics

text: WHAT IS KNOWN AND OBJECTIVE: There has been astounding progress made in the treatment of disease over recent years. This progress is particularly marked in cell therapy and in the personalization of therapy based on genetic insight, an approach known as genomic medicine. Our objective is to comment on the progress made in cell and genomic medicine against an historical backcloth of the search for rejuvenation. COMMENT: In 1741, close to seven decades after Antoine van Leeuwenhoek first saw his microscopic animalcules, Abraham Trembley, a tutor in Leiden, reported on an organism that could regenerate itself. The strange organism was thought to hold the secret of life. If it does, we have yet to prise the secret out. However, the ensuing study of cell programming and induced stem cells has shed considerable light on cellular development and provided new insights on the rejuvenative capacity of organisms. Inventive scientists have provided a deeper understanding of cell replication and, from this, developed new medicines for an increasing range of diseases. Targeted therapies, oligonucleotide therapy, therapeutic monoclonal antibodies and pharmacogenetics are all new therapeutic areas originating from the improved insights. More will surely follow. WHAT IS NEW AND CONCLUSION: Immortality is for the gods, but man's search for its elusive secrets, perhaps as old as man himself, will continue. Huge leaps have been made, and effective medicines have been developed from our improved insights into the mechanism of life. However, only the foolish will predict how far this new knowledge will lead us, and more particularly, at what speed new therapies will follow.


title: The Long‐Standing History of Corynebacterium Parvum, Immunity and Viruses

text: We report a review of all the experimental and clinical studies performed in the last 60 years on the antiviral activity of inactivated Corynebacterium Parvum (Cutibacterium acnes). This bacterium has been originally investigated and used for its oncolytic properties linked to immunomodulating activity, but the interest to successfully prevent and treat bacterial, fungal and viral infections and lethality, uprising the innate immunity barriers produced many experimental models and very few clinical studies. The dramatic defenceless situation due to impending CoViD‐19 pandemic claims to exhume and highlight this aspecific strategy in preventive and therapeutic settings; as a matter of fact no new or mutated virus can potentially escape to this strong innate immune surveillance strengthened by adequate C.parvum protocols. This article is protected by copyright. All rights reserved.


title: Investigation of soils affected by burnt hospital wastes in Nigeria using PIXE

text: Improper management of hospital waste has been reported to be responsible for several acute outbreaks like the severe acute respiratory syndrome (SARS). In spite of these challenges, hospital wastes are sometimes not properly handled in Nigeria. To date, there has not been an adequate study on the effect and fate of burnt hospital waste on agricultural soil. The effect of burnt hospital wastes on the agricultural soil was conducted on soils sampled around farm settlement near Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife, South West Nigeria. PIXE technique was employed with a 1.7 MV 5SDH Tandem Pelletron accelerator available at Centre for Energy Research and Development O.A.U Ile-Ife, Nigeria. Eleven elements- Si, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Zr and Pb were detected and their concentrations and enrichment factors determined. The presence of Pb and Cl at the elevated concentrations range of (77.8 ± 3.5 - 279.6 ± 97.6 and 102.2 ± 37.4 -167.2±17.43) ppm respectively in this study, is of serious health concern because of the agricultural practices in the neighborhoods of the study sites. There is a need for proper handling of hospital and other related hazardous wastes because of the possibility of such posing serious environmental pollution problems.


The COVID‐19 Pandemic: An Epidemiologic, Public Health, and Clinical Brief


title: The Role of Compassion and Ethics in Decision Making Regarding Access to Applied Behavior Analysis Services During the COVID-19 Crisis: A Response to Cox, Plavnick, and Brodhead

text: Cox, Plavnick, and Brodhead (2020, “A Proposed Process for Risk Mitigation During the COVID-19 Pandemic”) published a position statement in the emergency section of Behavior Analysis in Practice in response to the COVID-19 crisis. They argued against a blanket interpretation that in-person applied behavior analysis services for all patients should continue during the pandemic. They strongly argued that the risks of continued services are almost always prohibitive and that only in rare cases would the continuation of in-person services be warranted. Colombo, Wallace, and Taylor (2020, “An Essential Service Decisions Model for Applied Behavior Analytic Providers During Crisis”) soon thereafter published a response to the article pointing out the potential dangers associated with the position of the article by Cox et al. They included a detailed decision model to assist providers in making nuanced and informed data-based decisions that provide the opportunity to honor the ethical responsibility for not abandoning patients. We echo the importance of the Colombo et al. response and add points of response centered on balanced ethical decision making informed by compassionate family-centered care.


title: Viral and bacterial co-infection in severe pneumonia triggers innate immune responses and specifically enhances IP-10: a translational study

text: Mixed viral and bacterial infections are widely described in community-acquired pneumonia; however, the clinical implications of co-infection on the associated immunopathology remain poorly studied. In this study, microRNA, mRNA and cytokine/chemokine secretion profiling were investigated for human monocyte-derived macrophages infected in-vitro with Influenza virus A/H1N1 and/or Streptococcus pneumoniae. We observed that the in-vitro co-infection synergistically increased interferon-γ-induced protein-10 (CXCL10, IP-10) expression compared to the singly-infected cells conditions. We demonstrated that endogenous miRNA-200a-3p, whose expression was synergistically induced following co-infection, indirectly regulates CXCL10 expression by targeting suppressor of cytokine signaling-6 (SOCS-6), a well-known regulator of the JAK-STAT signaling pathway. Additionally, in a subsequent clinical pilot study, immunomodulators levels were evaluated in samples from 74 children (≤5 years-old) hospitalized with viral and/or bacterial community-acquired pneumonia. Clinically, among the 74 cases of pneumonia, patients with identified mixed-detection had significantly higher (3.6-fold) serum IP-10 levels than those with a single detection (P = 0.03), and were significantly associated with severe pneumonia (P < 0.01). This study demonstrates that viral and bacterial co-infection modulates the JAK-STAT signaling pathway and leads to exacerbated IP-10 expression, which could play a major role in the pathogenesis of pneumonia.


title: Applying next-generation sequencing to unravel the mutational landscape in viral quasispecies

text: Abstract Next-generation sequencing (NGS) has revolutionized the scale and depth of biomedical sciences. Because of its unique ability for the detection of sub-clonal variants within genetically diverse populations, NGS has been successfully applied to analyze and quantify the exceptionally-high diversity within viral quasispecies, and many low-frequency drug- or vaccine-resistant mutations of therapeutic importance have been discovered. Although many works have intensively discussed the latest NGS approaches and applications in general, none of them has focused on applying NGS in viral quasispecies studies, mostly due to the limited ability of current NGS technologies to accurately detect and quantify rare viral variants. Here, we summarize several error-correction strategies that have been developed to enhance the detection accuracy of minority variants. We also discuss critical considerations for preparing a sequencing library from viral RNAs and for analyzing NGS data to unravel the mutational landscape.


title: Activation of COX-2/PGE(2) Promotes Sapovirus Replication via the Inhibition of Nitric Oxide Production

text: Enteric caliciviruses in the genera Norovirus and Sapovirus are important pathogens that cause severe acute gastroenteritis in both humans and animals. Cyclooxygenases (COXs) and their final product, prostaglandin E(2) (PGE(2)), are known to play important roles in the modulation of both the host response to infection and the replicative cycles of several viruses. However, the precise mechanism(s) by which the COX/PGE(2) pathway regulates sapovirus replication remains largely unknown. In this study, infection with porcine sapovirus (PSaV) strain Cowden, the only cultivable virus within the genus Sapovirus, markedly increased COX-2 mRNA and protein levels at 24 and 36 h postinfection (hpi), with only a transient increase in COX-1 levels seen at 24 hpi. The treatment of cells with pharmacological inhibitors, such as nonsteroidal anti-inflammatory drugs or small interfering RNAs (siRNAs) against COX-1 and COX-2, significantly reduced PGE(2) production, as well as PSaV replication. Expression of the viral proteins VPg and ProPol was associated with activation of the COX/PGE(2) pathway. We observed that pharmacological inhibition of COX-2 dramatically increased NO production, causing a reduction in PSaV replication that could be restored by inhibition of nitric oxide synthase via the inhibitor N-nitro-l-methyl-arginine ester. This study identified a pivotal role for the COX/PGE(2) pathway in the regulation of NO production during the sapovirus life cycle, providing new insights into the life cycle of this poorly characterized family of viruses. Our findings also reveal potential new targets for treatment of sapovirus infection. IMPORTANCE Sapoviruses are among the major etiological agents of acute gastroenteritis in both humans and animals, but little is known about sapovirus host factor requirements. Here, using only cultivable porcine sapovirus (PSaV) strain Cowden, we demonstrate that PSaV induced the vitalization of the cyclooxygenase (COX) and prostaglandin E(2) (PGE(2)) pathway. Targeting of COX-1/2 using nonsteroidal anti-inflammatory drugs (NSAIDs) such as the COX-1/2 inhibitor indomethacin and the COX-2-specific inhibitors NS-398 and celecoxib or siRNAs targeting COXs, inhibited PSaV replication. Expression of the viral proteins VPg and ProPol was associated with activation of the COX/PGE(2) pathway. We further demonstrate that the production of PGE(2) provides a protective effect against the antiviral effector mechanism of nitric oxide. Our findings uncover a new mechanism by which PSaV manipulates the host cell to provide an environment suitable for efficient viral growth, which in turn can be a new target for treatment of sapovirus infection.


title: Turning off the tap: stopping tuberculosis transmission through active case-finding and prompt effective treatment

text: Summary To halt the global tuberculosis epidemic, transmission must be stopped to prevent new infections and new cases. Identification of individuals with tuberculosis and prompt initiation of effective treatment to rapidly render them non-infectious is crucial to this task. However, in settings of high tuberculosis burden, active case-finding is often not implemented, resulting in long delays in diagnosis and treatment. A range of strategies to find cases and ensure prompt and correct treatment have been shown to be effective in high tuberculosis-burden settings. The population-level effect of targeted active case-finding on reducing tuberculosis incidence has been shown by studies and projected by mathematical modelling. The inclusion of targeted active case-finding in a comprehensive epidemic-control strategy for tuberculosis should contribute substantially to a decrease in tuberculosis incidence.


title: Addressing the selectivity and toxicity of antiviral nucleosides

text: Nucleoside and nucleotide analogs have played significant roles in antiviral therapies and are valued for their impressive potency and high barrier to resistance. They have been approved for treatment of herpes simplex virus-1, HIV, HBV, HCV, and influenza, and new drugs are being developed for the treatment of RSV, Ebola, coronavirus MERS, and other emerging viruses. However, this class of compounds has also experienced a high attrition rate in clinical trials due to toxicity. In this review, we discuss the utility of different biochemical and cell-based assays and provide recommendations for assessing toxicity liability before entering animal toxicity studies.


Could pulmonary arterial hypertension patients be at a lower risk from severe COVID-19?


title: Phosphatidylinositol 4-Kinase IIIβ Is Required for Severe Acute Respiratory Syndrome Coronavirus Spike-mediated Cell Entry

text: Phosphatidylinositol kinases (PI kinases) play an important role in the life cycle of several viruses after infection. Using gene knockdown technology, we demonstrate that phosphatidylinositol 4-kinase IIIβ (PI4KB) is required for cellular entry by pseudoviruses bearing the severe acute respiratory syndrome-coronavirus (SARS-CoV) spike protein and that the cell entry mediated by SARS-CoV spike protein is strongly inhibited by knockdown of PI4KB. Consistent with this observation, pharmacological inhibitors of PI4KB blocked entry of SARS pseudovirions. Further research suggested that PI4P plays an essential role in SARS-CoV spike-mediated entry, which is regulated by the PI4P lipid microenvironment. We further demonstrate that PI4KB does not affect virus entry at the SARS-CoV S-ACE2 binding interface or at the stage of virus internalization but rather at or before virus fusion. Taken together, these results indicate a new function for PI4KB and suggest a new drug target for preventing SARS-CoV infection.


title: Hypereosinophilic paraneoplastic syndrome in a cat with intestinal T cell lymphosarcoma

text: A 10‐year‐old, neutered female, domestic shorthair cat was presented with a recent history of weight loss, polydipsia, diarrhea and vomiting. On physical examination, intestinal thickening and mesenteric lymph node enlargement were apparent. Clinical investigations revealed peripheral blood eosinophilia, eosinophilic abdominal effusion and eosinophilic mesenteric lymphadenitis. There was a temporary response to treatment with glucocorticoids but signs progressed and the cat was euthanased. On histology, there was eosinophilic infiltration and fibroplasia of intestine and mesenteric lymph nodes. Large aggregates of neoplastic round cells in the intestine and lymph nodes were identified as T lymphocytes using immunohistochemistry. A diagnosis of intestinal T cell lymphosarcoma was made. This case demonstrates that hypereosinophilic paraneoplastic syndrome may occur in cats with lymphosarcoma. Eosinophil chemotaxis may be a response to the production of interleukin‐5 by neoplastic lymphocytes.


title: Cystoviral RNA-directed RNA polymerases: Regulation of RNA synthesis on multiple time and length scales

text: Abstract P2, an RNA-directed RNA polymerase (RdRP), is encoded on the largest of the three segments of the double-stranded RNA genome of cystoviruses. P2 performs the dual tasks of replication and transcription de novo on single-stranded RNA templates, and plays a critical role in the viral life-cycle. Work over the last few decades has yielded a wealth of biochemical and structural information on the functional regulation of P2, on its role in the spatiotemporal regulation of RNA synthesis and its variability across the Cystoviridae family. These range from atomic resolution snapshots of P2 trapped in functionally significant states, in complex with catalytic/structural metal ions, polynucleotide templates and substrate nucleoside triphosphates, to P2 in the context of viral capsids providing structural insight into the assembly of supramolecular complexes and regulatory interactions therein. They include in vitro biochemical studies using P2 purified to homogeneity and in vivo studies utilizing infectious core particles. Recent advances in experimental techniques have also allowed access to the temporal dimension and enabled the characterization of dynamics of P2 on the sub-nanosecond to millisecond timescale through measurements of nuclear spin relaxation in solution and single molecule studies of transcription from seconds to minutes. Below we summarize the most significant results that provide critical insight into the role of P2 in regulating RNA synthesis in cystoviruses.


title: Chapter 12 Wine, food, and health

text: Abstract Wine has historically been associated with religious rights, used as a salubrious beverage, employed as a medication as well as a medicinal solvent, and consumed as a food accompaniment. It is the last use that is most intimately associated in the minds of most modern consumers. Despite this, there is little flavor commonality on which pairing could be based. The first section of the chapter examines this feature and wine's primary role as a palate cleanser and food condiment. The synergistic role of food and wine in suppressing each other's least pleasant attributes is also explained. The final section deals with the latest evidence relating to the many beneficial health effects of moderate wine consumption, shortfalls in the data, headache induction, dental erosion, and conditions under which wine intake is contraindicated.


title: Mouse models for the study of HCV infection and virus–host interactions

text: Hepatitis C virus (HCV) is a major cause of chronic liver disease including steatosis, cirrhosis and hepatocellular carcinoma. The development of transgenic mice expressing HCV proteins and the successful repopulation of SCID/Alb-uPA mice with human hepatocytes provides an important tool for unraveling virus–host interactions in vivo. Several of these mouse models exhibit aspects of HCV-related liver disease. Thus, these in vivo models play an important role to further understand the pathogenesis of HCV infection and to evaluate the pre-clinical safety and efficacy of new antiviral compounds against HCV. This review summarizes the most important mouse models currently used to study HCV pathogenesis and infection. Finally, the perspective of these models for future HCV research as well as the design of novel small animal models is discussed.


title: Letter to the Editor about the Beltrán‐Corbellini et al. publication :"Acute‐onset smell and taste disorders in the context of Covid‐19: a pilot multicenter PCR‐based case‐control study" (Eur J Neurol. 2020. doi: 10.1111/ene.14273.)

text: We read, with interest, the paper entitled "Acute‐onset smell and taste disorders in the context of Covid‐19: a pilot multicenter PCR‐based case‐control study". The authors observed that sudden loss of smell (LOS) was significantly more frequent in Covid‐19 patients (39%) than those infected with influenza (12%). These data corroborated the results of Yan et al.


title: Characterization of Kunjin virus RNA-dependent RNA polymerase: Reinitiation of synthesis in Vitro

text: Abstract RNA-dependent RNA polymerase (RDRP) activity was characterized in a cytoplasmic extract of Kunjin virus-infected Vero cells at 24 hr. The activity was influenced, possibly indirectly, by the length of prior treatment of infected cells with actinomycin D; however, 6 μg/ml actinomycin D and 10−5 M α-amanitin in the RDRP assay had no effect. The replication complex was membrane-bound and Mg2+ was essential for RDRP activity. Incorporation was more dependent on exogenous UTP and GTP than ATP or CTP. The specific activity was low, and rate of incorporation of GMP decreased as the period of assay was increased; however, incorporation of label lasted for at least 60 min. RNA products were fractionated by LiCl precipitation, and kinetic studies showed that the sequence of accumulation of label was the same as that observed in vivo, viz., RI → RF → 44 S RNA; limited reinitiation was also observed. This sequence of labeling also indicated that the in vitro RDRP activity was due to an enzyme capable of elongation, release, and reinitiation of Kunjin RNA synthesis and not merely end labeling or elongating preexisting RNA molecules. No labeled bands in urea-polyacrylamide gels were observed using extracts from mock-infected cells and hence the three RNA products of assays were readily identified in a single gel. The replication complex was still active after treatment with nonionic detergent, but no labeled 44 S RNA was detected in gels, even in the presence of RNasin in the assay which inhibited some nuclease activity. Antibodies to flavivirus-specific nonstructural proteins were preincubated with infected cell extracts in the presence and absence of detergent but no inhibition of RDRP activity was observed. However, anti-dsRNA plus detergent blocked activity by as much as 78% and label was found only in RF.


DGHM 2011


title: IgE reactivity to vaccine components in dogs that developed immediate-type allergic reactions after vaccination

text: Abstract Allergic reactions after vaccination are considered as an important practical problem in dogs; however, their immunological mechanism has not been well understood. The present study was designed to investigate the relationship between IgE reactivity to the vaccines and immediate-type allergic reactions after vaccination in dogs. Sera from 10 dogs that developed immediate-type allergic reactions such as circulatory collapse, cyanosis, dyspnea, facial edema, and vomiting within 1h after vaccination with non-rabies monovalent or combined vaccines and sera from 50 dogs that did not develop allergic reactions after vaccination were collected. Serum IgE reactivity to the injected vaccines was measured by fluorometric ELISA using a mouse monoclonal anti-dog IgE antibody. Then, IgE reactivity to fetal calf serum (FCS) and stabilizer proteins (gelatin, casein, and peptone) included in the vaccines was measured in sera that had high levels of IgE to the vaccines. Levels of serum specific IgE to the vaccines in dogs with immediate-type allergic reactions (59–4173 fluorescence units [FU], mean±S.D.: 992.5±1181.9 FU) were significantly higher than those in control dogs (38–192 FU, 92.4±43.3 FU) (P <0.001). Of the eight dogs that developed immediate-type allergic reactions and had high levels of serum specific IgE to the vaccines, seven had specific IgE directed to FCS. The IgE reactivity to the vaccines in sera from these dogs was almost completely inhibited by FCS. The other one dog had serum IgE directed to gelatin and casein included in the vaccine as stabilizers. The results obtained in this study suggest that immediate-type allergic reactions after vaccination in dogs were induced by type I hypersensitivity mediated by IgE directed to vaccine components. In addition, FCS, gelatin, and casein included in vaccines could be the causative allergens that induced immediate-type allergic reactions after vaccination in dogs.


title: Rash as a presenting complaint in a child with COVID‐19

text: Cutaneous manifestations are becoming increasingly well‐documented in adults with COVID‐19. There is now also a growing body of literature regarding skin involvement in children, with reports of papulovesicular, petechial and widespread macular and papular lesions, as well as chilblains (pernio). We describe the case of a thirteen‐year‐old boy with confirmed COVID‐19 in the United Kingdom who presented with skin findings localized to the plantar aspects of the feet, axillae and lower limbs. The morphology was predominantly maculopapular but also included petechiae and annular lesions.


title: An Online Observational Study of Patients With Olfactory and Gustory Alterations Secondary to SARS-CoV-2 Infection

text: Introduction: Since the beginning of the Covid-19 epidemic produced by SARS2-Cov virus, olfactory alterations have been observed at a greater frequency than in other coronavirus epidemics. While olfactory alterations may be observed in patients with rhinovirus, influenza virus, or parainfluenza virus infection, they are typically explained by nasal obstruction with mucus or direct epithelial damage; in the case of SARS-CoV-2, olfactory alterations may present without nasal congestion with mucus. We performed a study of patients presenting olfactory/gustatory alterations in the context of SARS-CoV-2 infection in order to contribute to the understanding of this phenomenon. Material and Methods: We performed a descriptive, cross-sectional, observational study of the clinical characteristics of olfactory/gustatory alterations using a self-administered, anonymous online questionnaire. Results: A total of 909 patients with SARS-CoV-2 infection and olfactory/gustatory alterations responded to the questionnaire in the 4-day data collection period; 824 cases (90.65%) reported simultaneous olfactory and gustatory involvement. Patients' responses to the questionnaire revealed ageusia (581, 64.1% of respondents), hypogeusia (256, 28.2%), dysgeusia (22, 2.4%), anosmia (752 82.8%), hyposmia (142, 15.6%), and dysosmia (8, 0.9%). Fifty-four percent (489) did not report concomitant nasal congestion or mucus. Conclusion: Olfactory alterations are frequent in patients with SARS-CoV-2 infection and is only associated with nasal congestion in half of the cases.


title: Threatwatch: Is the Saudi virus a new SARS?

text: The Middle Eastern coronavirus has started behaving disturbingly like SARS – making the search for undiagnosed cases increasingly vital


title: Epidemiology of and Risk Factors for Coronavirus Infection in Health Care Workers: A Living Rapid Review

text: BACKGROUND: Health care workers (HCWs) are at risk for severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection. PURPOSE: To examine the burden of SARS-CoV-2, SARS-CoV-1, and Middle Eastern respiratory syndrome (MERS)-CoV on HCWs and risk factors for infection, using rapid and living review methods. DATA SOURCES: Multiple electronic databases including the WHO Database of Publications on Coronavirus Disease and medRxiv preprint server (2003 through 27 March 2020, with ongoing surveillance through 24 April 2020), and reference lists. STUDY SELECTION: Studies published in any language reporting incidence of or outcomes associated with coronavirus infections in HCWs and studies on the association between risk factors (demographic characteristics, role, exposures, environmental and administrative factors, and personal protective equipment [PPE] use) and HCW infections. New evidence will be incorporated on an ongoing basis by using living review methods. DATA EXTRACTION: One reviewer abstracted data and assessed methodological limitations; verification was done by a second reviewer. DATA SYNTHESIS: 64 studies met inclusion criteria; 43 studies addressed burden of HCW infections (15 on SARS-CoV-2), and 34 studies addressed risk factors (3 on SARS-CoV-2). Health care workers accounted for a significant proportion of coronavirus infections and may experience particularly high infection incidence after unprotected exposures. Illness severity was lower than in non-HCWs. Depression, anxiety, and psychological distress were common in HCWs during the coronavirus disease 2019 outbreak. The strongest evidence on risk factors was on PPE use and decreased infection risk. The association was most consistent for masks but was also observed for gloves, gowns, eye protection, and handwashing; evidence suggested a dose–response relationship. No study evaluated PPE reuse. Certain exposures (such as involvement in intubations, direct patient contact, or contact with bodily secretions) were associated with increased infection risk. Infection control training was associated with decreased risk. LIMITATION: There were few studies on risk factors for SARS-CoV-2, the studies had methodological limitations, and streamlined rapid review methods were used. CONCLUSION: Health care workers experience significant burdens from coronavirus infections, including SARS-CoV-2. Use of PPE and infection control training are associated with decreased infection risk, and certain exposures are associated with increased risk. PRIMARY FUNDING SOURCE: World Health Organization.


title: A simple and rapid approach for screening of SARS-coronavirus genotypes: an evaluation study

text: BACKGROUND: The Severe Acute Respiratory Syndrome (SARS) was a newly emerged infectious disease which caused a global epidemic in 2002–2003. Sequence analysis of SARS-coronavirus isolates revealed that specific genotypes predominated at different periods of the epidemic. This information can be used as a footprint for tracing the epidemiology of infections and monitor viral evolution. However, direct sequencing analysis of a large number of clinical samples is cumbersome and time consuming. We present here a simple and rapid assay for the screening of SARS-coronavirus genotypes based on the use of fluorogenic oligonucleotide probes for allelic discrimination. METHODS: Thirty SARS patients were recruited. Allelic discrimination assays were developed based on the use of fluorogenic oligonucleotide probes (TaqMan). Genotyping of the SARS-coronavirus isolates obtained from these patients were carried out by the allelic discrimination assays and confirmed by direct sequencing. RESULTS: Genotyping based on the allelic discrimination assays were fully concordant with direct sequencing. All of the 30 SARS-coronavirus genotypes studied were characteristic of genotypes previously documented to be associated with the latter part of the epidemic. Seven of the isolates contained a previously reported major deletion but in patients not epidemiologically related to the previously studied cohort. CONCLUSION: We have developed a simple and accurate method for the characterization and screening of SARS-coronavirus genotypes. It is a promising tool for the study of epidemiological relationships between documented cases during an outbreak.


title: Anosmia: A marker of infection by the new corona virus

text: The diagnosis of Coronavirus Disease 2019 (COVID-19) is based on the identification of common symptoms such as fever, tiredness and dry cough. Anosmia and ageusia are also in fact symptoms of the infection with the new coronavirus and recently were considered as symptoms by the World Health Organization. In this case report we present the new onset anosmia during the COVID-19 pandemic. The patient, 31-year-old, reported olfactory and gustatory dysfunctions as initial symptoms of mild-to-moderate form of the COVID-19. Therefore, chemosensory dysfunctions should serve as a warning to health professionals as a possible marker of infection with the new corona virus.


title: Protecting Health Care Workers during the COVID-19 Coronavirus Outbreak –Lessons from Taiwan's SARS response

text: During major epidemic outbreaks, demand for health care workers grows even as the extreme pressures they face cause declining availability. We draw on Taiwan’s SARS experience to argue that a modified form of Traffic Control Bundling protects health care worker safety and by extension strengthens overall COVID-19 epidemic control.


title: Specific Recognition of Proteins by Array‐Bound Hexanucleotides

text: Maximum information density: The complete hexanucleotide sequence space can be immobilized on a chip. The binding of a number of proteins was profiled with this array of 4096 sequences, including fluorescently labeled forms of the HIV‐2 and HIV‐1 reverse transciptases (the consensus motif is shown in the picture). This new technique should aid in the search for pharmaceutical lead compunds.[Image: see text]


title: High-Value Plant Species Used for the Treatment of “Fever” by the Karen Hill Tribe People

text: The symptom “fever” is generally not itself a terminal condition. However, it does occur with common mild to severe ailments afflicting the world population. Several allopathic medicines are available to attenuate fever by targeting the pathogen or the symptom itself. However, many people in marginal civilizations are obligated to use locally grown medicinal plants due to limited access to common pharmaceuticals. The Karen ethnic group is the biggest ethnic minority group in the hill-tribes of Thailand. They utilise a vast repertoire of medicinal plant species. Since many modern drugs were discovered out of traditional therapies, it is possible to discover new allopathic drugs in the treatment of fever and associated pathogens from the Karen people. Thus, this study aims to identify and record the ethnomedicinal plants they used for the treatment of “fever”. The names of plants used by the Thai Karen people for the treatment of fever were mined from publications on ethnomedicinal uses. Useful plant species and families were identified using the Cultural Importance Index (CI). With the mined data, 125 plant species from 52 families were identified, distributed across 25 Karen villages. A chemical cross-examination of these species provided valuable insights into chemical classes worthy of further investigation in the context of fever and associated pathogens.


title: Chapter One Water Chemistry and Microbiology

text: Abstract Water is essential to life; therefore, providing access to safe and clean water is a major concern for health development at the local, national, and regional levels. Enhancing water quality by improving sanitation systems needs a profound understanding of water chemistry and microbiology. This chapter will give an overview of water chemistry and microbiology. The chapter will be divided into two main parts: the first one will discuss water chemistry in terms of its physical properties and the main chemical processes that takes place within water bodies, the main parameters that govern the water quality, and water quality management. This section will finally mark out the basics of water treatment processes. The second part will be devoted to water microbiology where the main concepts that are encountered when discussing water microbiology will be outlined. An overview will be given of the different types of microorganisms and their classification as well as the main processes they carry out in living cell such as metabolism, respiration, and microbial growth. A brief synopsis will be depicted about the main diseases caused by microorganisms that could be present in water bodies, the main microbial indicator, as well as the main analytical tests for microbial indicators. This section will also give a short overview about microbial control and the terms related to it such as disinfection. The final part of this section will delineate the role of microorganisms in wastewater treatment plants.


title: SARS-CoV-2 receptor ACE2 gene expression in small intestine correlates with age

text: Gastrointestinal symptoms are common in COVID-19 patients, especially in younger patients. Our hypothesis was that intestinal SARS-CoV-2 receptor ACE2 expression depends on patients’ age. We examined duodenal biopsies from 43 healthy human adults. ACE2 gene expression was directly correlated with age (Spearman’s r = 0.317, p = 0.039). With each year, duodenal ACE2 expression increased by 0.083 RU. The higher intestinal ACE2 mRNA expression in older patients may impact on their susceptibility to develop intestinal symptoms.


title: Effects of human T-cell leukemia virus type 1 (HTLV-1) p13 on mitochondrial K+ permeability: A new member of the viroporin family?

text: Abstract Human T-cell leukemia virus type-1 (HTLV-1) encodes a mitochondrial protein named p13. p13 mediates an inward K+ current in isolated mitochondria that leads to mitochondrial swelling, depolarization, increased respiratory chain activity and reactive oxygen species (ROS) production. These effects trigger the opening of the permeability transition pore and are dependent on the presence of K+ and on the amphipathic alpha helical domain of p13. In the context of cells, p13 acts as a sensitizer to selected apoptotic stimuli. Although it is not known whether p13 influences the activity of endogenous K+ channels or forms a channel itself, it shares some structural and functional analogies with viroporins, a class of small integral membrane proteins that form pores and alter membrane permeability.


title: Dysbiosis of upper respiratory tract microbiota in elderly pneumonia patients

text: Bacterial pneumonia is a major cause of morbidity and mortality in elderly. We hypothesize that dysbiosis between regular residents of the upper respiratory tract (URT) microbiome, that is balance between commensals and potential pathogens, is involved in pathogen overgrowth and consequently disease. We compared oropharyngeal microbiota of elderly pneumonia patients (n=100) with healthy elderly (n=91) by 16S-rRNA-based sequencing and verified our findings in young adult pneumonia patients (n=27) and young healthy adults (n=187). Microbiota profiles differed significantly between elderly pneumonia patients and healthy elderly (PERMANOVA, P<0.0005). Highly similar differences were observed between microbiota profiles of young adult pneumonia patients and their healthy controls. Clustering resulted in 11 (sub)clusters including 95% (386/405) of samples. We observed three microbiota profiles strongly associated with pneumonia (P<0.05) and either dominated by lactobacilli (n=11), Rothia (n=51) or Streptococcus (pseudo)pneumoniae (n=42). In contrast, three other microbiota clusters (in total n=183) were correlated with health (P<0.05) and were all characterized by more diverse profiles containing higher abundances of especially Prevotella melaninogenica, Veillonella and Leptotrichia. For the remaining clusters (n=99), the association with health or disease was less clear. A decision tree model based on the relative abundance of five bacterial community members in URT microbiota showed high specificity of 95% and sensitivity of 84% (89% and 73%, respectively, after cross-validation) for differentiating pneumonia patients from healthy individuals. These results suggest that pneumonia in elderly and young adults is associated with dysbiosis of the URT microbiome with bacterial overgrowth of single species and absence of distinct anaerobic bacteria. Whether the observed microbiome changes are a cause or a consequence of the development of pneumonia or merely coincide with disease status remains a question for future research.


title: Adaptive policies for use of physical distancing interventions during the COVID-19 pandemic

text: Policymakers need decision tools to determine when to use physical distancing interventions to maximize the control of COVID-19 while minimizing the economic and social costs of these interventions. We develop a pragmatic decision tool to characterize adaptive policies that combine real-time surveillance data with clear decision rules to guide when to trigger, continue, or stop physical distancing interventions during the current pandemic. In model-based experiments, we find that adaptive policies characterized by our proposed approach prevent more deaths and require a shorter overall duration of physical distancing than alternative physical distancing policies. Our proposed approach can readily be extended to more complex models and interventions.


title: Pregnant women voice their concerns and birth expectations during the COVID-19 pandemic in Italy

text: Abstract Background In March 2020, COVID-19 was declared to be a pandemic. While data suggests that COVID-19 is not associated with significant adverse health outcomes for pregnant women and newborns, the psychological impact on pregnant women is likely to be high. Aim The aim was to explore the psychological impact of the COVID-19 pandemic on Italian pregnant women, especially regarding concerns and birth expectations. Methods A cross-sectional online survey of pregnant women in Italy was conducted. Responses were analysed for all women and segregated into two groups depending on previous experience of pregnancy loss. Analysis of open text responses examined expectations and concerns before and after the onset of the pandemic. Findings Two hundred pregnant women responded to the first wave of the survey. Most (n = 157, 78.5%) had other children and 100 (50.0%) had a previous history of perinatal loss. ‘Joy’ was the most prevalent emotion expressed before COVID-19 (126, 63.0% before vs 34, 17.0% after; p < 0.05); fear was the most prevalent after (15, 7.5% before vs 98, 49.0% after; p < 0.05). Positive constructs were prevalent before COVID-19, while negative ones were dominant after (p < 0.05). Across the country, women were concerned about COVID-19 and a history of psychological disorders was significantly associated with higher concerns (p < 0.05). A previous pregnancy loss did not influence women’s concerns. Conclusions Women’s expectations and concerns regarding childbirth changed significantly as a result of the COVID-19 pandemic in Italy. Women with a history of psychological disorders need particular attention as they seem to experience higher levels of concern.


title: Antigen delivery systems for veterinary vaccine development Viral-vector based delivery systems

text: Abstract The recent advances in molecular genetics, pathogenesis and immunology have provided an optimal framework for developing novel approaches in the rational design of vaccines effective against viral epizootic diseases. This paper reviews most of the viral-vector based antigen delivery systems (ADSs) recently developed for vaccine testing in veterinary species, including attenuated virus and DNA and RNA viral vectors. Besides their usefulness in vaccinology, these ADSs constitute invaluable tools to researchers for understanding the nature of protective responses in different species, opening the possibility of modulating or potentiating relevant immune mechanisms involved in protection.


title: Bovine coronaviruses associated with enteric and respiratory diseases in Canadian dairy cattle display different reactivities to anti-HE monoclonal antibodies and distinct amino acid changes in their HE, S and ns4.9 protein

text: Abstract Bovine coronavirus isolates associated with recent outbreaks of respiratory disease in Ontario and Quebec dairy farms were compared to reference strains known to be responsible for neonatal calf diarrhea (NCD) or winter dysentery (WD) of adult cattle. In respect to their hemagglutinating properties and their higher RDE activities with rat erythrocytes, WDBCoV strains differed from NCDBCoV strains and respiratory bovine coronaviruses RBCoV strains. Serologically, three MAbs directed to the HE glycoprotein of the WDBCoV strain BCQ.2590 recognized two serogroups amongst NCDBCoV strains by hemagglutination inhibition, whereas only one of the MAbs failed to react toward three of the four RBCoV isolates tested. Sequencing analysis of the S (S1 portion), HE, ORF4 and ORF5 genes of BCoV isolates associated with different clinical syndromes indicated that neither insertions or deletions could explain their distinct tropism. For the HE glycoprotein, a total of 15 amino acids (aa) substitutions were identified by comparing field isolates to the prototype Mebus strain. Two specific proline substitutions were identified for virulent strains being located in the signal peptides (aa 5) and aa position 367; one specific aa change was revealed at position 66 for RBCoV field isolates. Analysis of the S1 portion of the S glycoprotein revealed a total of eight aa changes specific to enteropathogenic (EBCoV) strains and eight aa changes specific to RBCoV strains. For all BCoV isolates studied, the region located between the S and M genes (ORF4) apparently encodes for two non-structural (ns) proteins of 4.9 and 4.8 kDa. A specific non-sense mutation was identified for the nucleotide at position 88 of the putative 4.9 kDa protein gene of RBCoV isolates resulting in 29 rather that 43 aa residues. The ORF5, which encodes a 12.7 ns protein and the 9.5 kDa E protein, was highly conserved amongst the BCoV field isolates.


title: The nsp9 Replicase Protein of SARS-Coronavirus, Structure and Functional Insights

text: As part of a high-throughput structural analysis of SARS-coronavirus (SARS-CoV) proteins, we have solved the structure of the non-structural protein 9 (nsp9). This protein, encoded by ORF1a, has no designated function but is most likely involved with viral RNA synthesis. The protein comprises a single β-barrel with a fold previously unseen in single domain proteins. The fold superficially resembles an OB-fold with a C-terminal extension and is related to both of the two subdomains of the SARS-CoV 3C-like protease (which belongs to the serine protease superfamily). nsp9 has, presumably, evolved from a protease. The crystal structure suggests that the protein is dimeric. This is confirmed by analytical ultracentrifugation and dynamic light scattering. We show that nsp9 binds RNA and interacts with nsp8, activities that may be essential for its function(s).


How to transform a general hospital into an “infectious disease hospital” during the epidemic of COVID-19


title: Deficient IFN Signaling by Myeloid Cells Leads to MAVS-Dependent Virus-Induced Sepsis

text: The type I interferon (IFN) signaling response limits infection of many RNA and DNA viruses. To define key cell types that require type I IFN signaling to orchestrate immunity against West Nile virus (WNV), we infected mice with conditional deletions of the type I IFN receptor (IFNAR) gene. Deletion of the Ifnar gene in subsets of myeloid cells resulted in uncontrolled WNV replication, vasoactive cytokine production, sepsis, organ damage, and death that were remarkably similar to infection of Ifnar (−/−) mice completely lacking type I IFN signaling. In Mavs(−/−)×Ifnar(−/−) myeloid cells and mice lacking both Ifnar and the RIG-I-like receptor adaptor gene Mavs, cytokine production was muted despite high levels of WNV infection. Thus, in myeloid cells, viral infection triggers signaling through MAVS to induce proinflammatory cytokines that can result in sepsis and organ damage. Viral pathogenesis was caused in part by massive complement activation, as liver damage was minimized in animals lacking complement components C3 or factor B or treated with neutralizing anti-C5 antibodies. Disease in Ifnar (−/−) and CD11c Cre(+) Ifnar (f/f) mice also was facilitated by the proinflammatory cytokine TNF-α, as blocking antibodies diminished complement activation and prolonged survival without altering viral burden. Collectively, our findings establish the dominant role of type I IFN signaling in myeloid cells in restricting virus infection and controlling pathological inflammation and tissue injury.


title: Global status of Middle East respiratory syndrome coronavirus in dromedary camels: a systematic review

text: Dromedary camels have been shown to be the main reservoir for human Middle East respiratory syndrome (MERS) infections. This systematic review aims to compile and analyse all published data on MERS-coronavirus (CoV) in the global camel population to provide an overview of current knowledge on the distribution, spread and risk factors of infections in dromedary camels. We included original research articles containing laboratory evidence of MERS-CoV infections in dromedary camels in the field from 2013 to April 2018. In general, camels only show minor clinical signs of disease after being infected with MERS-CoV. Serological evidence of MERS-CoV in camels has been found in 20 countries, with molecular evidence for virus circulation in 13 countries. The seroprevalence of MERS-CoV antibodies increases with age in camels, while the prevalence of viral shedding as determined by MERS-CoV RNA detection in nasal swabs decreases. In several studies, camels that were sampled at animal markets or quarantine facilities were seropositive more often than camels at farms as well as imported camels vs. locally bred camels. Some studies show a relatively higher seroprevalence and viral detection during the cooler winter months. Knowledge of the animal reservoir of MERS-CoV is essential to develop intervention and control measures to prevent human infections.