title: Development of horse neutralizing immunoglobulin and immunoglobulin fragments against Junín virus

text: Abstract Argentine haemorrhagic fever (AHF) is a rodent-borne disease with a lethality as high as ~30%, which is caused by the New World arenavirus, Junín virus (JUNV). It was once a major epidemic in South America and puts millions of people in Argentina at risk. Here, we aimed to develop horse antibodies or antibody fragments against JUNV. Before preparing the horse antibodies, a strategy to efficiently generate horse antisera was established based on comparisons among immunogens and immunization methods in both mice and horses. Antisera against JUNV were finally obtained by vaccinating horses with vesicular stomatitis virus pseudotypes bearing JUNV GP. The horse antibodies IgG and F(ab’)2 were subsequently demonstrated to effectively neutralize vesicular stomatitis virus pseudotypes bearing JUNV GP and to show some cross-neutralization against pathogenic New World arenaviruses. Further research revealed that Asp123 on GP1 is an important site for the binding of antibodies targeting mainly JUNV GP1 for neutralization. Collectively, this study presents an efficient strategy to develop horse antisera against JUNV and provides GP1-specific horse antibodies as potential therapeutics for AHF.


title: Different virulence of porcine and porcine-like bovine rotavirus strains with genetically nearly identical genomes in piglets and calves

text: Direct interspecies transmissions of group A rotaviruses (RVA) have been reported under natural conditions. However, the pathogenicity of RVA has never been directly compared in homologous and heterologous hosts. The bovine RVA/Cow-tc/KOR/K5/2004/G5P[7] strain, which was shown to possess a typical porcine-like genotype constellation similar to that of the G5P[7] prototype RVA/Pig-tc/USA/OSU/1977/G5P9[7] strain, was examined for its pathogenicity and compared with the porcine G5P[7] RVA/Pig-tc/KOR/K71/2006/G5P[7] strain possessing the same genotype constellation. The bovine K5 strain induced diarrhea and histopathological changes in the small intestine of piglets and calves, whereas the porcine K71 strain caused diarrhea and histopathological changes in the small intestine of piglets, but not in calves. Furthermore, the bovine K5 strain showed extra-intestinal tropisms in both piglets and calves, whereas the porcine K71 strain had extra-intestinal tropisms in piglets, but not in calves. Therefore, we performed comparative genomic analysis of the K71 and K5 RVA strains to determine whether specific mutations could be associated with these distinct clinical and pathological phenotypes. Full-length sequencing analyses for the 11 genomic segments for K71 and K5 revealed that these strains were genetically nearly identical to each other. Two nucleotide mutations were found in the 5′ untranslated region (UTR) of NSP5 and the 3′ UTR of NSP3, and eight amino acid mutations in VP1-VP4 and NSP2. Some of these mutations may be critical molecular determinants for RVA virulence and/or pathogenicity.


title: Bioaerosol Sampling in Clinical Settings: A Promising, Noninvasive Approach for Detecting Respiratory Viruses

text: BACKGROUND. Seeking a noninvasive method to conduct surveillance for respiratory pathogens, we sought to examine the usefulness of 2 types of off-the-shelf aerosol samplers to detect respiratory viruses in Singapore. METHODS. In this pilot study, we ran the aerosol samplers several times each week with patients present in the patient waiting areas at 3 primary health clinics during the months of April and May 2016. We used a SKC BioSampler with a BioLite Air Sampling Pump (run for 60 min at 8 L/min) and SKC AirChek TOUCH personal air samplers with polytetrafluoroethylene Teflon filter cassettes (run for 180 min at 5 L/min). The aerosol specimens and controls were studied with molecular assays for influenza A virus, influenza B virus, adenoviruses, and coronaviruses. RESULTS. Overall, 16 (33.3%) of the 48 specimens indicated evidence of at least 1 respiratory pathogen, with 1 (2%) positive for influenza A virus, 3 (6%) positive for influenza B virus, and 12 (25%) positive for adenovirus. CONCLUSIONS. Although we were not able to correlate molecular detection with individual patient illness, patients with common acute respiratory illnesses were present during the samplings. Combined with molecular assays, it would suggest that aerosol sampling has potential as a noninvasive method for novel respiratory virus detection in clinical settings.


title: GAIA 3.0: Effects of the Coronavirus Disease 2019 (COVID-19) outbreak on sustainable development and future perspectives.

text: Abstract In the past decades, the frequency of epidemics with global importance has increased significantly. Only in the first two decades of the 21st Century, the world has witnessed the epidemics of Severe Acute Respiratory Syndromes, Zika virus, Ebola virus, Yellow Fever, the new COVID-19, besides the traditional influenza and other virus and bacteria. However, due to the high level of globalization, the large-scale population flow and the high reproductive rate of the virus, COVID-19 suddenly affected several countries, infecting hundreds of thousands of people and killing dozens of thousands in the fastest unprecedented crisis ever recorded, which also led to the fasted severe economic crisis on history. This perfect storm of social, economic and health catastrophes caused by the COVID-19 pandemics, directly threatens the world's livelihood and wellbeing and jeopardize the achievement of the United Nation's Sustainable Development Goals. Sustainable development is personified by Gaia's definition, implying limits for human activities and urging for a better use of technologies and resources. Although temporary, these changes in human actions set future trends for humanity, changing many aspects of our behavior, which include impacts in culture, technology, healthcare, economy, policy, education and the environment. Which may lead to a new enlightenment based on the need for global solidarity and an urge for the implementation of sustainable development pathways, finally creating a common agenda for the future of humanity as part of the Gaia, not above it.


Covid-19: Trump proposes tax cuts and improved health insurance, but millions are not covered.


title: Unmasking the Actual COVID-19 Case Count

text: This report presents a novel approach to estimate the number of COVID-19 cases, including undocumented infections, in the US, by combining CDC’s influenza-like illness surveillance data with aggregated prescription data. We estimated that the cumulative number of COVID-19 cases in the US by April 4 was above 2.5 million.


title: 1.47 Viruses and Virus-Like Particles in Biotechnology: Fundamentals and Applications ☆

text: This is a reprint of A. Roldão, A.C. Silva, M.C.M. Mellado, P.M. Alves, M.J.T. Carrondo, Viruses and Virus-Like Particles in Biotechnology: Fundamentals and Applications, Reference Module in Life Sciences, Elsevier, 2017.


Covid-19 in Immune-Mediated Inflammatory Diseases — Case Series from New York


title: Application of a Risk Analysis Tool to Middle East Respiratory Syndrome Coronavirus (MERS‐CoV) Outbreak in Saudi Arabia

text: The Grunow–Finke assessment tool (GFT) is an accepted scoring system for determining likelihood of an outbreak being unnatural in origin. Considering its high specificity but low sensitivity, a modified Grunow–Finke tool (mGFT) has been developed with improved sensitivity. The mGFT has been validated against some past disease outbreaks, but it has not been applied to ongoing outbreaks. This study is aimed to score the outbreak of Middle East respiratory syndrome coronavirus (MERS‐CoV) in Saudi Arabia using both the original GFT and mGFT. The publicly available data on human cases of MERS‐CoV infections reported in Saudi Arabia (2012–2018) were sourced from the FluTrackers, World Health Organization, Saudi Ministry of Health, and published literature associated with MERS outbreaks investigations. The risk assessment of MERS‐CoV in Saudi Arabia was analyzed using the original GFT and mGFT criteria, algorithms, and thresholds. The scoring points for each criterion were determined by three researchers to minimize the subjectivity. The results showed 40 points of total possible 54 points using the original GFT (likelihood: 74%), and 40 points of a total possible 60 points (likelihood: 67%) using the mGFT, both tools indicating a high likelihood that human MERS‐CoV in Saudi Arabia is unnatural in origin. The findings simply flag unusual patterns in this outbreak, but do not prove unnatural etiology. Proof of bioattacks can only be obtained by law enforcement and intelligence agencies. This study demonstrated the value and flexibility of the mGFT in assessing and predicting the risk for an ongoing outbreak with simple criteria.


title: Infections in Transplant Patients

text: Recipients of solid organ transplants (SOT) need primary care providers (PCPs) who are familiar with their unique needs and understand the lifelong infectious risks faced by SOT patients because of their need for lifelong immunosuppressive medications. SOT recipients can present with atypical and muted manifestations of infections, for which the knowledgable PCP will initiate a comprehensive evaluation. The goal of this article is to familiarize PCPs with the infectious challenges facing SOT patients. General concepts are reviewed, and a series of patient cases described that illustrate the specific learning points based on common presenting clinical symptoms.


title: Protecting intestinal epithelial cells against deoxynivalenol and E. coli damage by recombinant porcine IL-22

text: Abstract Pigs suffer enteritis induced by pathogenic bacteria infection and toxins in the moldy feed, which cause intestinal epithelial damage and diarrhea through the whole breeding cycle. Interleukin-22 (IL-22) plays a critical role in maintaining intestinal mucosal barrier function through repairing intestinal epithelial damage. However, little was known about the effects of IL-22 against apoptosis caused by toxins and infection of intestinal pathogens in the intestinal epithelium, especially in pigs. In this study, we had successfully used prokaryotic expression system to produce recombinant porcine interleukin-22. Meanwhile, purified rIL-22 could activate STAT3 signal pathway and have been demonstrated to be safe to IPEC-J2 cells by increasing E-cadherin expression, without proinflammatory cytokines changes. Furthermore, rIL-22 reversed apoptosis induced by deoxynivalenol (DON) and played a vital part in repairing the intestinal injury. We also found that rIL-22 stimulated epithelial cells to secrete pBD-1 against enterotoxigenic E. coli (ETEC) K88 infection, as well as alleviating apoptosis ratio. This study provided a theoretical basis for curing intestinal inflammation caused by ETEC infection and epithelial apoptosis induced by DON with rIL-22 in pigs.


title: In vitro assessment of the feline cell-mediated immune response against feline panleukopeniavirus, calicivirus and felid herpesvirus 1 using 5-bromo-2′-deoxyuridine labeling

text: Abstract In this study an in vitro assay was optimized to detect feline proliferating lymphocytes as an assessment for the cell-mediated immune response. For this purpose, 5-bromo-2′-deoxyuridine (BrdU) labeling was chosen because of its sensitivity and the possibility of further characterization of proliferating cells. The assay was optimized by selecting the best batch and concentration of fetal bovine serum, β-mercaptoethanol concentration, cell density, BrdU incubation time and antigen presenting cell type. Cats were vaccinated with the attenuated Nobivac vaccine Tricat and the peripheral blood lymphocyte proliferation responses were quantified upon in vitro restimulation with inactivated and infectious feline panleukopenia virus (FPV), feline calicivirus (FCV) and felid herpesvirus 1 (FeHV-1). Proliferation signals were detected with inactivated FeHV-1 in the CD8+ but not in the CD8− T lymphocyte population, with inactivated FCV and FPV in both CD8− and CD8+ T lymphocyte populations. Restimulation with infectious FCV caused significant proliferation in the CD8− T lymphocyte population only while infectious FPV and FeHV-1 seemed to suppress lymphocyte proliferation in both T cell populations. Additional IFN-γ quantification in the culture supernatant revealed a large correlation between the proliferation signals and IFN-γ production, indicating that BrdU labeling is a very reliable technique to assess and characterize feline lymphoproliferative responses to viral antigens in vitro.


title: Addressing COVID‐19 Drug Development with Artificial Intelligence

text: The emergence of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), the virus that led to the COVID‐19 (Coronavirus Disease 2019) pandemic, has resulted in substantial overburdening of healthcare systems as well as an economic crisis on a global scale. This has in turn resulted in widespread efforts to identify suitable therapies to address this aggressive pathogen. Therapeutic antibody and vaccine development are being actively explored, and a phase I clinical trial of mRNA‐1273 which is developed in collaboration between the National Institute of Allergy and Infectious Diseases and Moderna, Inc. is currently underway. Timelines for the broad deployment of a vaccine and antibody therapies have been estimated to be 12–18 months or longer. These are promising approaches that may lead to sustained efficacy in treating COVID‐19. However, its emergence has also led to a large number of clinical trials evaluating drug combinations composed of repurposed therapies. As study results of these combinations continue to be evaluated, there is a need to move beyond traditional drug screening and repurposing by harnessing artificial intelligence (AI) to optimize combination therapy design. This may lead to the rapid identification of regimens that mediate unexpected and markedly enhanced treatment outcomes.


title: Addressing General Surgery residents’ concerns in the early phase of the COVID-19 pandemic.

text: • General Surgery residents are preparing to be in the frontlines of the response against COVID-19. • They are ready to care for COVID-19 patients despite the risk of personal or familial harm. • Recommendations for their adequate preparedness include increased testing capacity, ensuring adequate personal protective equipment availability, and transitioning to a shift schedule in order to minimize exposure risk and prevent burnout.


title: Lessons from a one-year hospital-based surveillance of acute respiratory infections in Berlin- comparing case definitions to monitor influenza

text: BACKGROUND: Surveillance of severe acute respiratory infections (SARI) in sentinel hospitals is recommended to estimate the burden of severe influenza-cases. Therefore, we monitored patients admitted with respiratory infections (RI) in 9 Berlin hospitals from 7.12.2009 to 12.12.2010 according to different case definitions (CD) and determined the proportion of cases with influenza A(H1N1)pdm09 (pH1N1). We compared the sensitivity and specificity of CD for capturing pandemic pH1N1 cases. METHODS: We established an RI-surveillance restricted to adults aged ≤ 65 years within the framework of a pH1N1 vaccine effectiveness study, which required active identification of RI-cases. The hospital information-system was screened daily for newly admitted RI-patients. Nasopharyngeal swabs from consenting patients were tested by PCR for influenza-virus subtypes. Four clinical CD were compared in terms of capturing pH1N1-positives among hospitalized RI-patients by applying sensitivity and specificity analyses. The broadest case definition (CD1) was used for inclusion of RI-cases; the narrowest case definition (CD4) was identical to the SARI case definition recommended by ECDC/WHO. RESULTS: Over the study period, we identified 1,025 RI-cases, of which 283 (28%) met the ECDC/WHO SARI case definition. The percentage of SARI-cases among internal medicine admissions decreased from 3.2% (calendar-week 50-2009) to 0.2% (week 25-2010). Of 354 patients tested by PCR, 20 (6%) were pH1N1-positive. Two case definitions narrower than CD1 but -in contrast to SARI- not requiring shortness of breath yielded the largest areas under the Receiver-Operator-Curve. Heterogeneity of proportions of patients admitted with RI between hospitals was significant. CONCLUSIONS: Comprehensive surveillance of RI cases was feasible in a network of community hospitals. In most settings, several hospitals should be included to ensure representativeness. Although misclassification resulting from failure to obtain symptoms in the hospital information-system cannot be ruled out, a high proportion of hospitalized PCR-positive pH1N1-patients (45%) did not fulfil the SARI case-definition that included shortness of breath or difficulty breathing. Thus, to assess influenza-related disease burden in hospitals, broader, alternative case definitions should be considered.


A Journal in a Plague Year


title: Concerns regarding calculation of fatality rate of Chinese samples in COVID‐19 meta‐analysis

text: I read with interest the meta-analysis of patient clinical characteristics and fatality rates in COVID-19 from Chinese samples by Li and colleagues1 . I was surprised by the fatality rate of 7% which is much than higher than the value of 1.4% reported in a large study of Chinese patients by Guan et al (2020)2 . This article is protected by copyright. All rights reserved.


title: SARS‐CoV‐2 spike glycoprotein‐binding proteins expressed by upper respiratory tract bacteria may prevent severe viral infection

text: Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has become a major global challenge. The virus infects host cells using its spike glycoprotein (S‐protein) and has significantly higher infectivity and mortality rates among the aged population. Here, based on bioinformatic analysis, I provide evidence that some members of the upper respiratory tract (URT) commensal bacteria express viral S‐protein ‐binding proteins. Based on this analysis and available data showing a decline in the population of these bacteria in the elderly, I propose that some URT commensal bacteria hamper SARS‐CoV‐2 infectivity and that a decline in the population of these bacteria contributes to the severity of infection. Further studies should provide a better understanding of the interaction of URT bacteria and SARS‐CoV‐2, which may lead to new therapeutic approaches.


title: Isolation and sequence analysis of the complete H gene of canine distemper virus from domestic dogs in Henan Province, China

text: Eighteen canine distemper virus (CDV) isolates were obtained from clinical samples in Henan province, China, between 2012 and 2016. These viruses could not be recognized by 1A4, a monoclonal antibody specific for the H protein of CDV vaccine strains. The complete haemagglutinin (H) genes of all 18 isolates were sequenced, and phylogenetic analysis showed that they segregated into two clusters within the Asia-1 genotype. Moreover, the H genes of four viruses were found to lack a potential N-glycosylation site at position 309, which is the most conserved site within the Asia-1 genotype of CDV, and a novel potential N-glycosylation site (amino acids 517–519) was found in strain HL013, which has not been reported previously. These results will help in achieving a better understanding of the evolution of CDV in China. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00705-019-04298-7) contains supplementary material, which is available to authorized users.


On the relationship between science and reality in the time of COVID‐19


title: Dans les exacerbations de mucoviscidose, l’administration de tobramycine en une perfusion par jour ne parait pas plus toxique pour le rein ni moins efficace qu’en 3 perfusions par jour The TOPIC study group

text: Unknown


title: Coast-to-Coast Spread of SARS-CoV-2 during the Early Epidemic in the United States

text: Summary The novel coronavirus SARS-CoV-2 was first detected in the Pacific Northwest region of the United States in January 2020, with subsequent COVID-19 outbreaks detected in all 50 states by early March. To uncover the sources of SARS-CoV-2 introductions and patterns of spread within the United States, we sequenced nine viral genomes from early reported COVID-19 patients in Connecticut. Our phylogenetic analysis places the majority of these genomes with viruses sequenced from Washington state. By coupling our genomic data with domestic and international travel patterns, we show that early SARS-CoV-2 transmission in Connecticut was likely driven by domestic introductions. Moreover, the risk of domestic importation to Connecticut exceeded that of international importation by mid-March regardless of our estimated effects of federal travel restrictions. This study provides evidence of widespread sustained transmission of SARS-CoV-2 within the United States and highlights the critical need for local surveillance.


title: Prediction of the Epidemic Peak of Coronavirus Disease in Japan, 2020

text: The first case of coronavirus disease 2019 (COVID-19) in Japan was reported on 15 January 2020 and the number of reported cases has increased day by day. The purpose of this study is to give a prediction of the epidemic peak for COVID-19 in Japan by using the real-time data from 15 January to 29 February 2020. Taking into account the uncertainty due to the incomplete identification of infective population, we apply the well-known SEIR compartmental model for the prediction. By using a least-square-based method with Poisson noise, we estimate that the basic reproduction number for the epidemic in Japan is [Formula: see text] ([Formula: see text] CI, [Formula: see text] – [Formula: see text]) and the epidemic peak could possibly reach the early-middle summer. In addition, we obtain the following epidemiological insights: (1) the essential epidemic size is less likely to be affected by the rate of identification of the actual infective population; (2) the intervention has a positive effect on the delay of the epidemic peak; (3) intervention over a relatively long period is needed to effectively reduce the final epidemic size.


title: COVID-19-Related Stroke

text: The COVID-19 pandemic is associated with neurological symptoms and complications including stroke. There is hypercoagulability associated with COVID-19 that is likely a “sepsis-induced coagulopathy” and may predispose to stroke. The SARS-CoV-2 virus binds to angiotensin-converting enzyme 2 (ACE2) present on brain endothelial and smooth muscle cells. ACE2 is a key part of the renin angiotensin system (RAS) and a counterbalance to angiotensin-converting enzyme 1 (ACE1) and angiotensin II. Angiotensin II is proinflammatory, is vasoconstrictive, and promotes organ damage. Depletion of ACE2 by SARS-CoV-2 may tip the balance in favor of the “harmful” ACE1/angiotensin II axis and promote tissue injury including stroke. There is a rationale to continue to treat with tissue plasminogen activator for COVID-19-related stroke and low molecular weight heparinoids may reduce thrombosis and mortality in sepsis-induced coagulopathy.


title: Synthesis, antiviral, cytotoxic and cytostatic evaluation of N(1)-(phosphonoalkyl)uracil derivatives

text: ABSTRACT: A series of N (1)-(phosphonoalkyl)uracils was prepared in a two-step reaction sequence from ω-aminoalkylphosphonates and (E)-3-ethoxyacryloyl isocyanate followed by the uracil ring closure. Under standard conditions (NCS; NBS; I(2)/CAN) all N (1)-(phosphonoalkyl)uracils were transformed into the respective 5-halogeno derivatives to be later benzoylated at N3. All compounds were evaluated in vitro for activity against a broad variety of DNA and RNA viruses. One compound was slightly active against human cytomegalovirus in HEL cell cultures (EC(50) = 45 μM) while another showed weak activity against varicella-zoster virus (TK(+) VZV strain OKA and TK(−) VZV strain 07-1) with EC(50) = 43 and 53 µM, respectively. In addition, several compounds exhibited noticeable inhibitory effects on the proliferation of human cervical carcinoma cells (HeLa) at a concentration lower than 200 μM. GRAPHICAL ABSTRACT: [Image: see text]


title: Kaposi’s sarcoma: Etiology and pathogenesis, inducing factors, causal associations, and treatments: Facts and controversies

text: Abstract Kaposi's sarcoma (KS), an angioproliferative disorder, has a viral etiology and a multifactorial pathogenesis hinged on an immune dysfunction. The disease is multifocal, with a course ranging from indolent, with only skin manifestations to fulminant, with extensive visceral involvement. In the current view, all forms of KS have a common etiology in human herpesvirus (HHV)-8 infection, and the differences among them are due to the involvement of various cofactors. In fact, HHV-8 infection can be considered a necessary but not sufficient condition for the development of KS, because further factors (genetic, immunologic, and environmental) are required. The role of cofactors can be attributed to their ability to interact with HHV-8, to affect the immune system, or to act as vasoactive agents. In this contribution, a survey of the current state of knowledge on many and various factors involved in KS pathogenesis is carried out, in particular by highlighting the facts and controversies about the role of some drugs (quinine analogues and angiotensin-converting enzyme inhibitors) in the onset of the disease. Based on these assessments, it is possible to hypothesize that the role of cofactors in KS pathogenesis can move toward an effect either favoring or inhibiting the onset of the disease, depending on the presence of other agents modulating the pathogenesis itself, such as genetic predisposition, environmental factors, drug intake, or lymph flow disorders. It is possible that the same agents may act as either stimulating or inhibiting cofactors according to the patient’s genetic background and variable interactions. Treatment guidelines for each form of KS are outlined, because a unique standard therapy for all of them cannot be considered due to KS heterogeneity. In most cases, therapeutic options, both local and systemic, should be tailored to the patient’s peculiar clinical conditions.


title: Meat consumption: which are the current global risks? A review of recent (2010-2020) evidences

text: Meat consumption has been increasing since the 1960s, but especially from the 1980s decade to today. Although meat means an important source of nutrients, it is also evident that a great consumption of this source of proteins has also a negative environmental impact. Livestock production does not only have a negative influence on GHG emissions, but also on the water footprint, water pollution, and water scarcity. With respect to human health, in 2015 the International Agency for Research on Cancer (IARC) stated that red meat was a probable carcinogen to humans (Group 2A), while consumption of processed meat was carcinogenic to humans (Group 1). Most environmental contaminants (PCDD/Fs, PCBs, PBDEs, PCNs, etc.) that are frequently found in meats are highly soluble in fats. Therefore, avoiding ingesting fats from red meats and meat products, doubtless would help in the prevention, not only of the well-known cardiovascular diseases derived of fats consumption, but also of certain kinds of cancers, mainly colorectal cancer. On the other hand, consumption of meat – especially wild meat – is related to virus infections, as many viruses have been found in wild meat trade markets. Based on the scientific literature here reviewed, we have noted that the results of the investigations conducted after the statement of the IARC, have corroborated the recommendation of reducing significantly the consumption of red meats and meat products. In turn, the reduction of meat consumption should contribute to the reduction of GHG emissions and their considerable impact on global warming and climate change. It seems evident that human dietary habits regarding meat consumption in general, and red meats and wild meats in particular, should be significantly modified downward, as much and as soon as possible.


title: MERS epidemiological investigation to detect potential mode of transmission in the 178th MERS confirmed case in Pyeongtaek, Korea

text: Most cases of Middle East Respiratory Syndrome (MERS) infection in Korea (outbreak: May 11-July 4, 2015) occurred in hospital settings, with uncertain transmission modes in some cases. We performed an in-depth investigation epidemiological survey on the 178th case to determine the precise mode of transmission. A 29- year-old man living in Pyeongtaek presented on June 16 with a febrile sensation, chills, and myalgia. Upon confirmatory diagnosis on June 23, he was treated in an isolation room and discharged on July 2 after cure. An epidemiological investigation of all possible infection routes indicated two likely modes of transmission: exposure to MERS in Pyeongtaek St. Mary’s Hospital during a visit to his hospitalized father (May 18-29), and infection through frequent contact with his father between the latter’s referral to Pyeongtaek Good Samaritan Bagae Hospital for treatment without confirmatory diagnosis until his death (May 29-June 6). Although lack of clear proof or evidence to the contrary does not allow a definitive conclusion, all other possibilities could be excluded by epidemiological inferences. While it is impossible to trace back the modes of transmission of all cases in a large-scale outbreak, case-by-case tracking and isolation of infected individuals and those in close contact with them is important in preventing the spread. Efforts should be made to establish a methodology for rapid tracking of all possible contacts and elimination-based identification of the precise modes of transmission.


title: Chlamydia pneumoniae and Exacerbations of Asthma in Adults

text: Background Chlamydia pneumoniae is a frequent causative agent of acute respiratory disease and has been recently reported as a possible case of asthma. Objective We assessed the prevalence of C. pneumoniae infections in adult patients with acute exacerbations of asthma. Methods One hundred sixty-eight adult patients with acute exacerbations of asthma and 108 control subjects matched for age, sex, and smoking status were studied. Nasopharyngeal swab specimens were obtained from all subjects and analyzed by isolation in cell culture and polymerase chain reaction (PCR) test for C. pneumoniae. Serum samples were also obtained and tested for C. pneumoniae-specific antibodies by the microimmunofluorescence test. Results C. pneumoniae was isolated from two (1.2%) asthma patients and none from controls and detected by PCR from nine (5.4%) cases and one (0.9%) control. Both culture positive specimens were also positive in PCR. Further, serologic evidence of acute C. pneumoniae infection was present in 15 (8.9%) of asthma patients and in three (2.8%) of controls (P = .048). The prevalence of C. pneumoniae-specific IgG and IgA was significantly higher in asthma cases than in controls (IgG >/= 1:16; 85.1% versus 67.6%, P = .001; IgA ≥ 1:16: 47.6% versus 16.7%, P < .001). Mean titer of IgG and IgA was also significantly greater in asthma cases than in controls (IgG: 38.8 versus 18.1, P = .0001; IgA: 17.2 versus 6.1, P = .0001). Conclusion Our data suggest that C. pneumoniae infection may trigger acute exacerbations of adult asthma.


title: Preoperative assessment organization in the time of the outbreak COVID-19

text: • Maintaining the health of clinical workforce during COVID-19 pandemic is a top priority to avoiding collapse of health care system. • Following extraordinary measures could greatly reduce the risk of cross-contamination for both staff and non-infected patients. • Airway assessment is a critical time for the consultant anesthetist. • Projecting the airway examination via a mirror placed in front of the patient can be a useful idea to minimizing viral contamination.


title: The Baculoviruses Occlusion‐Derived Virus: Virion Structure and Function

text: Publisher Summary Baculoviruses play an important ecological role regulating the size of insect populations. For many years, baculoviruses have been applied as targeted biocontrol agents against forestry and agriculture pests. Baculovirus insecticides are effective against insect pests such as velvetbean caterpillar (Anticarsia gemmatalis), cotton bollworm (Helicoverpa zea), and gypsy moth (Lymantria dispar). Baculoviruses are transmitted to insects by the oral route mediated by the occlusion-derived virus (ODV). The ODV is also specialized to exploit the insect midgut that is one of the most extreme biological environments where the viruses are subject to caustic pH and digestive proteases. The molecular biology of the ODV reveals new frontiers in protein chemistry. Finally, ODVs establishes infection in insect gut tissues that are virtually nonsupportive to virus replication and which are continuously sloughed away. ODVs carry with them a battery of proteins that enable them to rapidly exploit and harness these unstable cells for virus replication.


title: The lactate dehydrogenase-elevating virus capsid protein is a nuclear–cytoplasmic protein

text: Arteriviruses replicate in the cytoplasm and do not require the nucleus function for virus multiplication in vitro. However, nucleocapsid (N) protein of two arteriviruses, porcine reproductive respiratory syndrome virus and equine arteritis virus, has been observed to localize in the nucleus and nucleolus of virus-infected and N-gene-transfected cells in addition to their normal cytoplasmic distribution. In the present study, the N protein of lactate dehydrogenase-elevating virus (LDV) of mice was examined for nuclear localization. The subcellular localization of LDV-N was determined by tagging N with enhanced green fluorescence protein (EGFP) at the N- and C-terminus. Both N-EGFP and EGFP-N fusion proteins localized to the nucleus and nucleolus of gene-transfected cells. Labeled N also accumulated in the perinuclear region, the site of virus replication. The LDV-N sequence contains a putative ‘pat4’-type nuclear localization signal (NLS) consisting of 38-KKKK. To determine its functional significance, a series of deletion constructs of N were generated and individually expressed in cells. The results showed that the ‘pat4’ NLS was essential for nuclear translocation. In addition, the LDV-N interacted with the importin-α and -β proteins, suggesting that the LDV-N nuclear localization may occur via the importin-mediated nuclear transport pathway. These results provide further evidence for the nuclear localization of N as a common feature within the arteriviruses.


title: Chancen und Einsatzmöglichkeiten von Telemedizin in der Hals‑, Nasen- und Ohrenheilkunde bei der Bekämpfung von SARS-COV-2: Narratives Review

text: INTRODUCTION: The COVID-19 pandemic is bringing healthcare systems worldwide to the brink of collapse. One reason for this is the rapidly increasing number of new infections. On the other hand, the high sickness rates of doctors and nurses, particularly in ENT medicine, are aggravating the situation. Telemedicine can be a useful tool to reduce the number of physician–patient contacts. This could break infection chains and minimize the risk of infection for physicians. METHODS: To prepare the review, a selective literature search was conducted at www.pubmed.com using the relevant English technical terms for telemedicine and ENT. In addition, research was conducted at www.news.google.com on current developments of the COVID-19 pandemic with the search terms “telemedicine” and “COVID-19.” RESULTS: Telemedicine can be helpful in direct contact with patients as well as in the conciliar support of general practitioners. The available studies show that, on average, more than 50% of medical consultations could be carried out telemedically. Both physicians and patients rate the use of telemedicine positively. Neither image quality nor the handling of the technology are relevant obstacles to a reliable diagnosis. Patients indicated that the telemedical consultation did not last longer than a traditional consultation. Patients also highlighted the faster and better availability of medical care through telemedicine. CONCLUSION: Telemedicine can make a decisive contribution to coping with the current COVID-19 pandemic. Furthermore, the establishment of telemedicine can help us to become better prepared for future pandemics.


COVID‐19 a short‐term challenge, telewound a lifetime change


title: Saturation mutagenesis genome engineering of infective ΦX174 bacteriophage via unamplified oligo pools and golden gate assembly

text: Here we present a novel protocol for the construction of saturation single-site—and massive multi-site—mutant libraries of a bacteriophage. We segmented the ΦX174 genome into 14 non-toxic and non-replicative fragments compatible with golden gate assembly. We next used nicking mutagenesis with oligonucleotides prepared from unamplified oligo pools with individual segments as templates to prepare near-comprehensive single-site mutagenesis libraries of genes encoding the F capsid protein (421 amino acids scanned) and G spike protein (172 amino acids scanned). Libraries possessed greater than 99% of all 11,860 programmed mutations. Golden Gate cloning was then used to assemble the complete ΦX174 mutant genome and generate libraries of infective viruses. This protocol will enable reverse genetics experiments for studying viral evolution and, with some modifications, can be applied for engineering of therapeutically relevant bacteriophages with larger genomes.


title: Adaptive Gene Amplification As an Intermediate Step in the Expansion of Virus Host Range

text: The majority of recently emerging infectious diseases in humans is due to cross-species pathogen transmissions from animals. To establish a productive infection in new host species, viruses must overcome barriers to replication mediated by diverse and rapidly evolving host restriction factors such as protein kinase R (PKR). Many viral antagonists of these restriction factors are species specific. For example, the rhesus cytomegalovirus PKR antagonist, RhTRS1, inhibits PKR in some African green monkey (AGM) cells, but does not inhibit human or rhesus macaque PKR. To model the evolutionary changes necessary for cross-species transmission, we generated a recombinant vaccinia virus that expresses RhTRS1 in a strain that lacks PKR inhibitors E3L and K3L (VVΔEΔK+RhTRS1). Serially passaging VVΔEΔK+RhTRS1 in minimally-permissive AGM cells increased viral replication 10- to 100-fold. Notably, adaptation in these AGM cells also improved virus replication 1000- to 10,000-fold in human and rhesus cells. Genetic analyses including deep sequencing revealed amplification of the rhtrs1 locus in the adapted viruses. Supplying additional rhtrs1 in trans confirmed that amplification alone was sufficient to improve VVΔEΔK+RhTRS1 replication. Viruses with amplified rhtrs1 completely blocked AGM PKR, but only partially blocked human PKR, consistent with the replication properties of these viruses in AGM and human cells. Finally, in contrast to AGM-adapted viruses, which could be serially propagated in human cells, VVΔEΔK+RhTRS1 yielded no progeny virus after only three passages in human cells. Thus, rhtrs1 amplification in a minimally permissive intermediate host was a necessary step, enabling expansion of the virus range to previously nonpermissive hosts. These data support the hypothesis that amplification of a weak viral antagonist may be a general evolutionary mechanism to permit replication in otherwise resistant host species, providing a molecular foothold that could enable further adaptations necessary for efficient replication in the new host.


title: TNFα inhibitor may be effective for severe COVID-19: learning from toxic epidermal necrolysis

text: Increased inflammatory cytokines [such as tumor necrosis factor alpha (TNFα) and interleukin-6 (IL-6)] are observed in COVID-19 patients, especially in the severe group. The phenomenon of a cytokine storm may be the central inducer of apoptosis of alveolar epithelial cells, which leads to rapid progression in severe group patients. Given the similarities of clinical features and pathogenesis between toxic epidermal necrolysis (TEN) and COVID-19, we hypothesize that the application of etanercept, an inhibitor of TNFα, could attenuate disease progression in severe group COVID-19 patients by suppressing systemic auto-inflammatory responses. The reviews of this paper are available via the supplemental material section.


title: Transmission of Mouse Minute Virus (MMV) but Not Mouse Hepatitis Virus (MHV) Following Embryo Transfer with Experimentally Exposed In Vivo-Derived Embryos(1)

text: The present study investigated the presence and location of fluorescent microspheres having the size of mouse hepatitis virus (MHV) and of mouse minute virus (MMV) in the zona pellucida (ZP) of in vivo-produced murine embryos, the transmission of these viruses by embryos during embryo transfer, and the time of seroconversion of recipients and pups. To this end, fertilized oocytes and morulae were exposed to different concentrations of MMVp for 16 h, while 2-cell embryos and blastocysts were coincubated for 1 h. In addition, morulae were exposed to MHV-A59 for 16 h. One group of embryos was washed, and the remaining embryos remained unwashed before embryo transfer. Serological analyses were performed by means of ELISA to detect antibodies to MHV or MMV in recipients and in progeny on Days 14, 21, 28, 42, and 63 and on Days 42, 63, 84, 112, 133, and 154, respectively, after embryo transfer. Coincubation with a minimum of 10(5)/ml of fluorescent microspheres showed that particles with a diameter of 20 nm but not 100 nm crossed the ZP of murine blastocysts. Washing generally led to a 10-fold to 100-fold reduction of MMVp. Washed MMV-exposed but not MHV-exposed embryos led to the production of antibodies independent of embryonic stage and time of virus exposure. Recipients receiving embryos exposed to a minimum of 10(7) mean tissue culture infective dose (TCID(50))/ml of MHV-A59 and 10(2) TCID(50)/ml of MMVp seroconverted by Day 42 after embryo transfer. The results indicate that MMV but not MHV can be transmitted to recipients even after washing embryos 10 times before embryo transfer.


title: Association between infectious diseases and type 1 diabetes: a case‐crossover study

text: Background: To investigate the role of infectious diseases in the development of type 1 diabetes, this study estimated the relative risks of type 1 diabetes immediately after infectious diseases. Research design and methods: A case‐crossover design was employed. Information on infectious diseases during 407 d before the onset of type 1 diabetes was collected from medical records and parents' interviews for 260 patients in Chinese type 1 diabetes registry. The frequency of infectious diseases in 42 d before the onset of type 1 diabetes was compared with either the usual frequency of infectious diseases over the past year or the actual frequency of infectious diseases in a comparable 42‐d control period. Results: Forty‐eight (18%) patients were reported to have infectious diseases during this period based on medical records and interviews with parents. The relative risk of type 1 diabetes onset was markedly elevated to 10.1 (5.6, 17.9) immediately after infectious diseases, suggesting the role of infections as a precipitator. The relative risk decreased gradually before and after 42 d and was similar between male and female patients. Conclusion: The results showed that infectious diseases are associated with a large and transient increase in the risk of type 1 diabetes during 42 d after the infection.


title: The Burden of Disease due to COVID-19 in Korea Using Disability-Adjusted Life Years

text: BACKGROUND: The world is currently experiencing a pandemic of coronavirus disease 2019 (COVID-19). In Korea, as in other countries, the number of confirmed cases and deaths due to COVID-19 have been rising. This study aimed to calculate the burden of disease due to COVID-19 in Korea. METHODS: We used data on confirmed cases and deaths due to COVID-19 between January 20 and April 24, 2020 provided by the Korea Centers for Disease Control and Prevention, the local governments and the public media to determine disability-adjusted life years (DALYs) by sex and age. Morbidity was estimated directly among the confirmed, cured, and fatal cases. Disability weights were adopted from previous similar causes on the severity of COVID-19 for the years of life lived with disability (YLDs). The years of life lost (YLLs) were calculated using the standard life expectancy from the 2018 life tables for each sex and age. RESULTS: The YLDs were higher in females (155.2) than in males (105.1), but the YLLs were higher in males (1,274.3) than in females (996.4). The total disease burden attributable to COVID-19 in Korea during the study period, was estimated to be 2,531.0 DALYs, and 4.930 DALYs per 100,000 population. The YLDs and the YLLs constituted 10.3% and 89.7% of the total DALYs, respectively. The DALYs per 100,000 population were highest in people aged ≥ 80 years, followed by those aged 70–79, 60–69, and 50–59 years, but the incidence was the highest in individuals aged 20–29 years. CONCLUSION: This study provided the estimates of DALYs due to COVID-19 in Korea. Most of the disease burden from COVID-19 was derived from YLL; this indicates that decision-makers should focus and make an effort on reducing fatality for preparing the second wave of COVID-19.


Ordering Effect Induced by SARS-CoV Fusion Peptides on Membranes Containing Negatively Charged Lipids Might be Important to Membrane Fusion


title: International Circumpolar Surveillance, An Arctic Network for the Surveillance of Infectious Diseases

text: Peoples of the Arctic and sub-Arctic regions live in social and physical environments that differ substantially from those of their more southern-dwelling counterparts. The cold northern climate keeps people indoors, amplifying the effects of household crowding, smoking, and inadequate ventilation on person-to-person spread of infectious disease. The emergence of antimicrobial drug resistance among bacterial pathogens, the reemergence of tuberculosis, the entrance of HIV into Arctic communities, and the specter of pandemic influenza or the sudden emergence and introduction of new viral pathogens such as severe acute respiratory syndrome are of increasing concern to residents, governments, and public health authorities. The International Circumpolar Surveillance system is a network of hospital, public health agencies, and reference laboratories throughout the Arctic linked together to collect, compare, and share uniform laboratory and epidemiologic data on infectious diseases and assist in the formulation of prevention and control strategies.


SARS-CoV-2 Testing: Trials and Tribulations


title: Genetic drift of human coronavirus OC43 spike gene during adaptive evolution

text: Coronaviruses (CoVs) continuously threaten human health. However, to date, the evolutionary mechanisms that govern CoV strain persistence in human populations have not been fully understood. In this study, we characterized the evolution of the major antigen-spike (S) gene in the most prevalent human coronavirus (HCoV) OC43 using phylogenetic and phylodynamic analysis. Among the five known HCoV-OC43 genotypes (A to E), higher substitution rates and dN/dS values as well as more positive selection sites were detected in the S gene of genotype D, corresponding to the most dominant HCoV epidemic in recent years. Further analysis showed that the majority of substitutions were located in the S1 subunit. Among them, seven positive selection sites were chronologically traced in the temporal evolution routes of genotype D, and six were located around the critical sugar binding region in the N-terminal domain (NTD) of S protein, an important sugar binding domain of CoV. These findings suggest that the genetic drift of the S gene may play an important role in genotype persistence in human populations, providing insights into the mechanisms of HCoV-OC43 adaptive evolution.


title: Prevalence of MERS-CoV Nasal Carriage and Compliance With the Saudi Health Recommendations Among Pilgrims Attending the 2013 Hajj

text: Background. Annually, Saudi Arabia is the host of the Hajj mass gathering. We aimed to determine the Middle East respiratory syndrome coronavirus (MERS-CoV) nasal carriage rate among pilgrims performing the 2013 Hajj and to describe the compliance with the Saudi Ministry of Health vaccine recommendations. Method. Nasopharyngeal samples were collected from 5235 adult pilgrims from 22 countries and screened for MERS-CoV using reverse transcriptase–polymerase chain reaction. Information regarding the participants' age, gender, country of origin, medical conditions, and vaccination history were obtained. Results. The mean age of the screened population was 51.8 years (range, 18–93 years) with a male/female ratio of 1.17:1. MERS-CoV was not detected in any of the samples tested (3210 pre-Hajj and 2025 post-Hajj screening). According to the vaccination documents, all participants had received meningococcal vaccination and the majority of those from at-risk countries were vaccinated against yellow fever and polio. Only 22% of the pilgrims (17.5% of those ≥65 years and 36.3% of diabetics) had flu vaccination, and 4.4% had pneumococcal vaccination. Conclusion. There was no evidence of MERS-CoV nasal carriage among Hajj pilgrims. While rates of compulsory vaccinations uptake were high, uptake of pneumococcal and flu seasonal vaccinations were low, including among the high-risk population.


Provisions for Obstetrics and Gynaecology - letter to the editor on “Impact of the coronavirus (COVID-19) pandemic on surgical practice - Part 2 (surgical prioritisation)”: A correspondence


PPG Donating 80,000 Masks to Support Coronavirus Relief Efforts


title: Roles of natural killer cells in antiviral immunity

text: Natural killer (NK) cells are important in immune defense against virus infections. This is predominantly considered a function of rapid, innate NK-cell killing of virus-infected cells. However, NK cells also prime other immune cells through the release of interferon gamma (IFN-γ) and other cytokines. Additionally, NK cells share features with long-lived adaptive immune cells and can impact disease pathogenesis through the inhibition of adaptive immune responses by virus-specific T and B cells. The relative contributions of these diverse and conflicting functions of NK cells in humans are poorly defined and likely context-dependent, thereby complicating the development of therapeutic interventions. Here we focus on the contributions of NK cells to disease in diverse virus infections germane to human health.


title: Experimental Cross-Species Infection of Common Marmosets by Titi Monkey Adenovirus

text: Adenoviruses are DNA viruses that infect a number of vertebrate hosts and are associated with both sporadic and epidemic disease in humans. We previously identified a novel adenovirus, titi monkey adenovirus (TMAdV), as the cause of a fulminant pneumonia outbreak in a colony of titi monkeys (Callicebus cupreus) at a national primate center in 2009. Serological evidence of infection by TMAdV was also found in a human researcher at the facility and household family member, raising concerns for potential cross-species transmission of the virus. Here we present experimental evidence of cross-species TMAdV infection in common marmosets (Callithrix jacchus). Nasal inoculation of a cell cultured-adapted TMAdV strain into three marmosets produced an acute, mild respiratory illness characterized by low-grade fever, reduced activity, anorexia, and sneezing. An increase in virus-specific neutralization antibody titers accompanied the development of clinical signs. Although serially collected nasal swabs were positive for TMAdV for at least 8 days, all 3 infected marmosets spontaneously recovered by day 12 post-inoculation, and persistence of the virus in tissues could not be established. Thus, the pathogenesis of experimental inoculation of TMAdV in common marmosets resembled the mild, self-limiting respiratory infection typically seen in immunocompetent human hosts rather than the rapidly progressive, fatal pneumonia observed in 19 of 23 titi monkeys during the prior 2009 outbreak. These findings further establish the potential for adenovirus cross-species transmission and provide the basis for development of a monkey model useful for assessing the zoonotic potential of adenoviruses.


title: Risk factors for pandemic H1N1 2009 infection in healthcare personnel of four general hospitals

text: Summary To characterize an outbreak of pandemic H1N1 2009 among healthcare personnel (HCP), we conducted a cross-sectional survey of HCP who had worked in four general hospitals during the outbreak. More than one-quarter of responding HCP (27.6%) had influenza-like illness (ILI) during the outbreak. The estimated infection rate of pandemic H1N1 2009 was 9.1% in the study of HCP. Independent risk factors for ILI were female gender, <40 years of age, the presence of chronic diseases associated with influenza complications, having family members with ILI or pandemic H1N1 2009, and working in influenza outpatient, influenza inpatient, non-influenza outpatient or emergency departments. During the outbreak of pandemic H1N1 2009, HCP frequently had ILI or the influenza infection. The development of the influenza infection in HCP was associated with some of their baseline characteristics, occupational risk factors, and non-occupational ones during the outbreak.


title: Targeting of the Nasal Mucosa by Japanese Encephalitis Virus for Non-Vector-Borne Transmission

text: The mosquito-borne Japanese encephalitis virus (JEV) causes severe central nervous system diseases and cycles between Culex mosquitoes and different vertebrates. For JEV and some other flaviviruses, oronasal transmission is described, but the mode of infection is unknown. Using nasal mucosal tissue explants and primary porcine nasal epithelial cells (NEC) at the air-liquid interface (ALI) and macrophages as ex vivo and in vitro models, we determined that the nasal epithelium could represent the route of entry and exit for JEV in pigs. Porcine NEC at the ALI exposed to with JEV resulted in apical and basolateral virus shedding and release of monocyte recruiting chemokines, indicating infection and replication in macrophages. Moreover, macrophages stimulated by alarmins, including interleukin-25, interleukin-33, and thymic stromal lymphopoietin, were more permissive to the JEV infection. Altogether, our data are important to understand the mechanism of non-vector-borne direct transmission of Japanese encephalitis virus in pigs. IMPORTANCE JEV, a main cause of severe viral encephalitis in humans, has a complex ecology composed of a mosquito-waterbird cycle and a cycle involving pigs, which amplifies virus transmission to mosquitoes, leading to increased human cases. JEV can be transmitted between pigs by contact in the absence of arthropod vectors. Moreover, virus or viral RNA is found in oronasal secretions and the nasal epithelium. Using nasal mucosa tissue explants and three-dimensional porcine nasal epithelial cells cultures and macrophages as ex vivo and in vitro models, we determined that the nasal epithelium could be a route of entry as well as exit for the virus. Infection of nasal epithelial cells resulted in apical and basolateral virus shedding and release of monocyte recruiting chemokines and therefore infection and replication in macrophages, which is favored by epithelial-cell-derived cytokines. The results are relevant to understand the mechanism of non-vector-borne direct transmission of JEV.


title: Crystallization and diffraction analysis of the SARS coronavirus nsp10–nsp16 complex

text: To date, the SARS coronavirus is the only known highly pathogenic human coronavirus. In 2003, it was responsible for a large outbreak associated with a 10% fatality rate. This positive RNA virus encodes a large replicase polyprotein made up of 16 gene products (nsp1–16), amongst which two methyltransferases, nsp14 and nsp16, are involved in viral mRNA cap formation. The crystal structure of nsp16 is unknown. Nsp16 is an RNA-cap AdoMet-dependent (nucleoside-2′-O-)-methyltransferase that is only active in the presence of nsp10. In this paper, the expression, purification and crystallization of nsp10 in complex with nsp16 are reported. The crystals diffracted to a resolution of 1.9 Å resolution and crystal structure determination is in progress.


title: Vaccine efficacy of a cell lysate with recombinant baculovirus-expressed feline infectious peritonitis (FIP) virus nucleocapsid protein against progression of FIP

text: Abstract The Type II feline infectious peritonitis virus (FIPV) infection of feline macrophages is enhanced by a monoclonal antibody (MAb) to the S protein of FIPV. This antibody-dependent enhancement (ADE) activity increased with the MAb that showed a neutralizing activity with feline kidney cells, suggesting that there was a distinct correlation between ADE activity and the neutralizing activity. The close association between enhancing and neutralizing epitopes is an obstacle to developing a vaccine containing only neutralizing epitopes without enhancing epitopes. In this study, we immunized cats with cell lysate with recombinant baculovirus-expressed N protein of the Type I FIPV strain KU-2 with an adjuvant and investigated its preventive effect on the progression of FIP. Cats immunized with this vaccine produced antibodies against FIPV virion-derived N protein but did not produce virus-neutralizing antibodies. A delayed type hypersensitivity skin response to N protein was observed in these vaccinated cats, showing that cell mediated immunity against the FIPV antigen was induced. When these vaccinated cats were challenged with a high dose of heterologous FIPV, the survival rate was 75% (6/8), while the survival rate in the control group immunized with SF-9 cell-derived antigen was 12.5% (1/8). This study showed that immunization with the cell lysate with baculovirus-expressed N protein was effective in preventing the progression of FIP without inducing ADE of FIPV infection in cats.


title: Putative contributions of circadian clock and sleep in the context of SARS-CoV-2 infection

text: Circadian deregulation and poor or insufficient sleep may facilitate COVID19 infection and severity


title: Novel algorithm for accelerated electroanatomic mapping and prediction of earliest activation of focal cardiac arrhythmias using mathematical optimization

text: Background Premature beats (PBs) are a common finding in patients suffering from structural heart disease, but they can also be present in healthy individuals. Catheter ablation represents a suitable therapeutic approach. However, the exact localization of the origin can be challenging, especially in cases of low PB burden during the procedure. Objective The aim of this study was to develop an automated mapping algorithm on the basis of the hypothesis that mathematical optimization would significantly accelerate the localization of earliest activation. Methods The algorithm is based on iterative regression analyses. When acquiring local activation times (LATs) within a 3-dimensional anatomic map of the corresponding heart chamber, this algorithm is able to identify that exact position where a next LAT measurement adds maximum information about the predicted site of origin. Furthermore, on the basis of the acquired LAT measurements, the algorithm is able to predict earliest activation with high accuracy. Results A systematic retrospective analysis of the mapping performance comparing the operator with simulated search processes by the algorithm within 17 electroanatomic maps of focal spreading arrhythmias revealed a highly significant reduction of necessary LAT measurements from 55 ± 8.8 to 10 ± 0.51 (n = 17; P < .0001). Conclusion On the basis of mathematical optimization, we developed an algorithm that is able to reduce the number of LAT measurements necessary to locate the site of earliest activation. This algorithm might significantly accelerate the mapping procedure by guiding the operator to the optimal position for the next LAT measurement. Furthermore, the algorithm would be able to predict the site of origin with high accuracy early during the mapping procedure.


title: Insertion of Introns: A Strategy to Facilitate Assembly of Infectious Full Length Clones

text: Some DNA fragments are difficult to clone in Escherichia coli by standard methods. It has been speculated that unintended transcription and translation result in expression of proteins that are toxic to the bacteria. This problem is frequently observed during assembly of infectious full-length virus clones. If the clone is constructed for transcription in vivo, interrupting the virus sequence with an intron can solve the toxicity problem. The AU-rich introns generally contain many stop codons, which interrupt translation in E. coli, while the intron sequence is precisely eliminated from the virus sequence in the plant nucleus. The resulting RNA, which enters the cytoplasm, is identical to the virus sequence and can initiate infection.


Electron Microscopic Investigations in COVID-19: Not all Crowns Are Coronas


title: Response to infections in persons with asthma and atopic disease: epiphenomenon or reflection of host susceptibility

text: Associations between respiratory infections and asthma inception and exacerbations are well established. Infant respiratory syncytial virus and rhinovirus infections are known to be associated with an increased risk of asthma development, and among children with prevalent asthma, 85% of asthma exacerbations are associated with viral infections. However, the exact nature of this relationship remains unclear. Is the increase in severity of infections an epiphenomenon, meaning respiratory infections just appear more severe in individuals with underlying respiratory disease, or instead a reflection of altered host susceptibility among persons with asthma and atopic disease? The main focus of this review is to summarize the available levels of evidence supporting or refuting the notion that persons with asthma or atopic disease have an altered susceptibility to selected pathogens, as well as discussing the biological mechanism(s) that might explain such associations. Finally, we will outline areas in need of further research, as understanding the relationships between infections and asthma has important implications for both asthma prevention and treatment, including potential new pathways that might target host immune response to select pathogens.


title: The current status of viral immunology

text: Abstract Many aspects of viral immunity, ranging from the molecular and cellular studies of the interaction between viruses and host cells in vitro and the crystalline structures of the MHC and peptides, to the regulation of pathogenesis in experimental animals and humans were discussed at a recent meeting.


title: Functional characterization of a broad and potent neutralizing monoclonal antibody directed against outer membrane protein (OMP) of Salmonella typhimurium

text: In the present study, we have generated a murine monoclonal antibody (mAb) named Sal-06 by using the crude outer membrane protein preparation of Salmonella enteric subsp. enterica serovar Typhimurium ATCC 14028 strain as antigen. Sal-06mAb belonging to IgG1 isotype demonstrated broad cross-reactivity to standard and isolated strains of genus Salmonella and others such as Escherichia coli, Klebsiella pneumonia, and Proteus mirabilis. Cross-reactivity across several bacterial genera indicated that the epitopes reactive to Sal-06mAb are conserved among these members. Neutralizing effects of Sal-06mAb on Salmonella growth and survival was evaluated in vitro using bacteriostatic and bactericidal activity with and without complement and bacterial invasion inhibition assay. Sal-06mAb demonstrated a bacteriostatic effect on the growth of S. typhimurium ATCC 14028 strain which is both time and concentration (of mAb) dependent. It was also found that the bacterial growth inhibition was complement independent. When the bacterial cells were preincubated with Sal-06mAb, it reduced the adherence and invasion of bacterial cells into A549 epithelial cell line. This was confirmed by CFU count analysis, phase contrast, and fluorescence microscopy. Scanning electron microscope (SEM) imaging confirmed the antimicrobial effects of Sal-06mAb on S. typhimurium ATCC 14028. The development of broadly reactive and cross protective Sal-06mAb opens new possibilities for immunotherapy of sepsis caused by Gram-negative Enterobacteriaceae members.


title: Discrepancies in viral gastroenteritis diagnosis: an unusual dual reovirus–adenovirus infection case

text: Abstract Background : A variable rate of false-positive results may be observed with commercial assays for the detection of rotavirus and adenovirus antigen in stool specimens, depending on the quality of the reagents and the presence of potentially interfering substances in stool samples. Objective : The present report analyse the discrepant results that could be obtained by the commercially available diagnostic tests and that can mask the reliable viral diagnosis. Study design : One fecal sample was collected from a hospitalized child aged 6 months with acute watery diarrhea and dehydration. The fecal specimen was processed the same day for the rotavirus and adenovirus antigen detection. Results : The sample was positive for rotavirus antigen by one-step membrane test based on immunochromatographic assays (ICA) and enzyme immunoassays (EIA) monoclonal test but it was negative by an EIA polyclonal test, polyacrylamide gel electrophoresis (PAGE) and RT-PCR assays. In the other hand, the sample was positive for adenovirus antigen by ICA and EIA adenovirus type 40/41. Finally, the sample showed by PAGE an electrophoretic profile resembled that of reovirus. Conclusion : The use of a wide repertory of diagnosis tests allowed to reach an unusual reovirus–adenovirus type 40/41 dual infection. This case also point out the potential participation of reovirus in the ethiology of the diarrhea illness.


title: How to safely and sustainably reorganise a large general radiography service facing the COVID-19 pandemic

text: Abstract Objectives This paper aims to share our experience in reorganising our general radiography service during the coronavirus disease (COVID-19) pandemic from the viewpoint of a large tertiary referral medical centre. Key findings Re-organization of radiographic manpower, patient segregation, and modification of routine radiographic practices are key measures to help radiographic services deal with the COVID-19 pandemic. More specifically, emphasis on deploying more mobile radiographic units, segregating equipment, developing consistent image acquisition workflows, and strict adherence to infection control protocols are paramount to minimize the possibility of in-hospital transmission and ensure a safe environment for both patients and staff. Streamlining communication channels between leadership and ground staff allows quick dissemination of information to ultimately facilitate safe provision of services. Conclusion COVID-19 has drastically altered the way general radiography teams provide services. The institution of several key measures will allow hospitals to safely and sustainably provide radiographic services. To date, there have been zero incidences of radiographer healthcare worker transmission within our institution during the course of work. Implication for practice Radiographers are facing the challenge of providing high-quality services while simultaneously minimizing pathogen exposure to staff and patients. Our experience may lend support to other radiographic services respond to the COVID-19 outbreak and serve as a blueprint for future infectious disease outbreak contingency plans.


How Do You Boost Resilience during the Current COVID-19 Pandemic in Your Practice?


Introduction to immunology at The University of Iowa


title: Non-suppurative Meningoencephalitis of Unknown Origin in Cats and Dogs: an Immunohistochemical Study

text: Summary Non-suppurative meningoencephalitis of unknown cause is a frequent finding in dogs and cats. Fifty-three dogs and 33 cats with non-suppurative meningoencephalitis of unknown aetiology were examined immunohistochemically for 18 different infectious agents, including viruses, bacteria and prion proteinSc. In 14 (26%) of the dogs and 13 (39%) of the cats a causative agent was identified in the central nervous system (CNS), two dogs and one cat giving positive results for two infectious agents simultaneously. The study revealed infections with known causative agents (porcine herpes virus 1, feline infectious peritonitis virus, Escherichia coli) and a new disease pattern of parvovirus infection in the CNS of dogs and cats. Infection of the CNS with feline leukaemia virus was found in a cat. Five dogs and four cats gave positive results for West Nile virus (WNV) antigen. In one dog, canine parainfluenza virus antigen was detected in the brain. Four dogs and four cats gave positive results for encephalomyocarditis virus (EMCV). The significance of the detection of WNV and EMCV antigen requires further study. The aetiology remained undetermined in 39 dogs (74%) and 20 cats (61%). Although it is possible that non-infectious causes play a more important role than previously thought, infections with hitherto unrecognized agents cannot be ruled out.


title: Facile route for N 1-aryl benzotriazoles from diazoamino arynes via CuI-mediated intramolecular N-arylation

text: Abstract A facile and high-yielding protocol for diverse benzotriazoles through intramolecular N-arylation of different o-chloro-1,2,3-benzotriazenes using CuI/Cs2CO3 has been developed.


Acute olfactory loss is specific for Covid-19 at the Emergency Department.


COVID‐19 pandemics and oral health care for older adults


title: 2 Application of Plastics in Medical Devices and Equipment

text: This chapter will present a look at the medical device market with a particular focus on the materials of construction of devices and what we can expect in new products looking ahead. A deeper look at some other trends that have an effect on the direction of the medical device industry will be done. Finally, consideration will be given to a number of global factors that can have dramatic effects on our industry.


title: Chapter 33 Avian Coronavirus: Case of Infectious Bronchitis Virus Pathogenesis, Diagnostic Approaches, and Phylogenetic Relationship Among Emerging Strains in Middle East and North Africa Regions

text: Abstract Infectious bronchitis (IB) virus (IBV) is a very dynamic and evolving virus, causing severe diseases in poultry with significant economic consequences to the poultry industry worldwide, affecting chickens of all ages and possessing a major threat to the poultry industry. IBV is well studied worldwide; nevertheless, in Middle East and North Africa (MENA) regions, epidemiological and scientific data are poor and not updated. In spite of regular vaccination with Massachusetts (Mass) strain has been available to control IB for many decades in MENA regions, which it is most commonly used, the protection is not occurred. However, the continuity of the spread of IBV has shown the emergence of multiple variant strains due to the frequent point of mutations and recombination events in the S1 gene hypervariable regions. All the reported IBV variants circulate in the form of several serotypes/genotypes that can be partially or poorly neutralized by current vaccines. This emergence remains a problem for the poultry industry and vaccine manufacturers. Therefore the objective of this present chapter, which is reported for the first time, aims to establish phylogenic relationship among emerging strains in MENA regions.


title: Control of Infectious Diseases in the Era of European Clinical Microbiology Laboratory Consolidation: New Challenges and Opportunities for the Patient and for Public Health Surveillance

text: Many new innovative diagnostic approaches have been made available during the last 10 years with major impact on patient care and public health surveillance. In parallel, to enhance the cost-effectiveness of the clinical microbiology laboratories (CMLs), European laboratory professionals have streamlined their organization leading to amalgamation of activities and restructuring of their professional relationships with clinicians and public health specialists. Through this consolidation process, an operational model has emerged that combines large centralized clinical laboratories performing most tests on one high-throughput analytical platform connected to several distal laboratories dealing locally with urgent analyses at near point of care. The centralization of diagnostic services over a large geographical region has given rise to the concept of regional-scale “microbiology laboratories network.” Although the volume-driven cost savings associated with such laboratory networks seem self-evident, the consequence(s) for the quality of patient care and infectious disease surveillance and control remain less obvious. In this article, we describe the range of opportunities that the changing landscape of CMLs in Europe can contribute toward improving the quality of patient care but also the early detection and enhanced surveillance of public health threats caused by infectious diseases. The success of this transformation of health services is reliant on the appropriate preparation in terms of staff, skills, and processes that would be inclusive of stakeholders. In addition, rigorous metrics are needed to set out more concrete laboratory service performance objectives and assess the expected benefits to society in terms of saving lives and preventing diseases.


title: COVID-19 im Alter — eine komplexe Herausforderung: Corona-Pandemie 2020

text: Alte und multimorbide Patienten sind besonders vulnerabel für einen schweren Verlauf von COVID-19. Die Maßnahmen zur Eindämmung der Pandemie und die Anpassung der Versorgungsstrukturen wirken sich massiv auf die ambulante geriatrische und die Notfallversorgung aus, sowie auf den akutstationären Bereich bis hin zu geriatrischer Rehabilitation und stationärer Pflege.


SARS-Cov2 Clinical Diagnostics: Academic Scientists Take on the COVID-19 Pandemic


Albumin and total bilirubin for severity and mortality in coronavirus disease 2019 patients


title: Dysregulated expression of T cell immunoglobulin and mucin domain 3 is associated with the disease severity and the outcome of patients with spontaneous intracerebral hemorrhage

text: Abstract Objectives We aimed to investigate the expression of T cell immunoglobulin and mucin domain 3 (Tim-3) on peripheral blood cells in spontaneous intracerebral hemorrhage (ICH) patients and to analyze its clinical significance. Design and methods Tim-3 expression on peripheral immunocytes from ICH patients and healthy volunteers was measured by flow cytometry. The correlation between Tim-3 expression and the clinical indices was estimated using linear regression. Results Tim-3 expressions on peripheral CD3+ T cells and CD8+ T cells in ICH patients are significantly downregulated, while Tim-3 expressions on CD14+ monocytes and CD16+CD56+ NK cells are increased. Furthermore, Tim-3 expression on peripheral CD8+ T cells was negatively correlated with the inflammatory response, the disease severity and the outcome of ICH patients. However, there was no relationship between Tim-3 expression and blood glucose concentration. Conclusions Altered expression of Tim-3 might play an important role in the pathogenesis of ICH, demonstrating that Tim-3 might be a novel candidate molecule for prognosis evaluation of ICH patients.


title: Household financial vulnerability in Indonesia: Measurement and determinants()

text: This study assesses the level of financial vulnerability of Indonesian households using data from the Household’s Balance Sheet Survey (Survei Neraca Rumah Tangga/SNRT) 2016 and 2017. The SNRT are micro-unit of household data that contains information on preferences and behavior. Through both objective and subjective measurements of the Household Financial Vulnerability Index (FVI), we find that the financial vulnerability of Indonesian households is not only strongly influenced by income factors, but also by finance-related behavioral characteristics and several socio-economic factors. As a consistency and robustness check, we also estimate econometric models using the Indonesian Family Life Survey (IFLS) panel data for the periods 1993, 1997, 2000, 2007 and 2014. Our study then conclude that the level of household financial vulnerability decreased in 2017. Moreover, the study suggests that we should carefully monitor the behaviour of middle income group as they contribute significantly to the household financial vulnerability in Indonesia.


title: Exchange proteins directly activated by cAMP (EPACs): Emerging therapeutic targets

text: Exchange proteins directly activated by cAMP (EPACs) are critical cAMP-dependent signaling pathway mediators. The discovery of EPAC proteins has significantly facilitated understanding on cAMP-dependent signaling pathway and efforts along this line open new avenues for developing novel therapeutics for cancer, diabetes, heart failure, inflammation, infections, neurological disorders and other human diseases. Over the past decade, important progress has been made in the identification of EPAC agonists, antagonists and their biological and pharmacological applications. In this review, we briefly summarize recently reported novel functions of EPACs and the discovery of their small molecule modulators. The challenges and future perspectives are also discussed.


title: Antiviral effects of Glycyrrhiza species

text: Historical sources for the use of Glycyrrhiza species include ancient manuscripts from China, India and Greece. They all mention its use for symptoms of viral respiratory tract infections and hepatitis. Randomized controlled trials confirmed that the Glycyrrhiza glabra derived compound glycyrrhizin and its derivatives reduced hepatocellular damage in chronic hepatitis B and C. In hepatitis C virus‐induced cirrhosis the risk of hepatocellular carcinoma was reduced. Animal studies demonstrated a reduction of mortality and viral activity in herpes simplex virus encephalitis and influenza A virus pneumonia. In vitro studies revealed antiviral activity against HIV‐1, SARS related coronavirus, respiratory syncytial virus, arboviruses, vaccinia virus and vesicular stomatitis virus. Mechanisms for antiviral activity of Glycyrrhiza spp. include reduced transport to the membrane and sialylation of hepatitis B virus surface antigen, reduction of membrane fluidity leading to inhibition of fusion of the viral membrane of HIV‐1 with the cell, induction of interferon gamma in T‐cells, inhibition of phosphorylating enzymes in vesicular stomatitis virus infection and reduction of viral latency. Future research needs to explore the potency of compounds derived from licorice in prevention and treatment of influenza A virus pneumonia and as an adjuvant treatment in patients infected with HIV resistant to antiretroviral drugs. Copyright © 2007 John Wiley & Sons, Ltd.


Modified mask for aesthetic procedures on face during COVID‐19 era: chiseling our armamentarium


title: A nascent peptide is required for ribosomal bypass of the coding gap in bacteriophage T4 gene 60

text: Abstract Bacteriophage T4 DNA topoisomerase gene 60 contains a 50 nucleotide untranslated region within the coding sequence of its mRNA. Translational bypass of this sequence by elongating ribosomes has been postulated for the mode of synthesis of an 18 kd polypeptide specified by the split coding segments. Ribosome bypass of the untranslated region also occurs when a segment of gene 60 is fused to lacZ and expressed in E. coli. The efficiency of bypass in these gene 60-lacZ fusions approaches 100%. Here, mutations that delete, insert, or substitute nucleotides from gene 60-lacZ fusions are examined. Essential features necessary for high level gap bypass emerging from this analysis are a cis-acting nascent peptide sequence, a short duplication bordering the gap, and a stop codon contained in a stem-loop structure at the 5′ junction of the gap.


title: A Novel Approach to Clustering Genome Sequences Using Inter-nucleotide Covariance

text: Classification of DNA sequences is an important issue in the bioinformatics study, yet most existing methods for phylogenetic analysis including Multiple Sequence Alignment (MSA) are time-consuming and computationally expensive. The alignment-free methods are popular nowadays, whereas the manual intervention in those methods usually decreases the accuracy. Also, the interactions among nucleotides are neglected in most methods. Here we propose a new Accumulated Natural Vector (ANV) method which represents each DNA sequence by a point in ℝ(18). By calculating the Accumulated Indicator Functions of nucleotides, we can further find an Accumulated Natural Vector for each sequence. This new Accumulated Natural Vector not only can capture the distribution of each nucleotide, but also provide the covariance among nucleotides. Thus global comparison of DNA sequences or genomes can be done easily in ℝ(18). The tests of ANV of datasets of different sizes and types have proved the accuracy and time-efficiency of the new proposed ANV method.


title: Exposure to Ozone Modulates Human Airway Protease/Antiprotease Balance Contributing to Increased Influenza A Infection

text: Exposure to oxidant air pollution is associated with increased respiratory morbidities and susceptibility to infections. Ozone is a commonly encountered oxidant air pollutant, yet its effects on influenza infections in humans are not known. The greater Mexico City area was the primary site for the spring 2009 influenza A H1N1 pandemic, which also coincided with high levels of environmental ozone. Proteolytic cleavage of the viral membrane protein hemagglutinin (HA) is essential for influenza virus infectivity. Recent studies suggest that HA cleavage might be cell-associated and facilitated by the type II transmembrane serine proteases (TTSPs) human airway trypsin-like protease (HAT) and transmembrane protease, serine 2 (TMPRSS2), whose activities are regulated by antiproteases, such as secretory leukocyte protease inhibitor (SLPI). Based on these observations, we sought to determine how acute exposure to ozone may modulate cellular protease/antiprotease expression and function, and to define their roles in a viral infection. We utilized our in vitro model of differentiated human nasal epithelial cells (NECs) to determine the effects of ozone on influenza cleavage, entry, and replication. We show that ozone exposure disrupts the protease/antiprotease balance within the airway liquid. We also determined that functional forms of HAT, TMPRSS2, and SLPI are secreted from human airway epithelium, and acute exposure to ozone inversely alters their expression levels. We also show that addition of antioxidants significantly reduces virus replication through the induction of SLPI. In addition, we determined that ozone-induced cleavage of the viral HA protein is not cell-associated and that secreted endogenous proteases are sufficient to activate HA leading to a significant increase in viral replication. Our data indicate that pre-exposure to ozone disrupts the protease/antiprotease balance found in the human airway, leading to increased influenza susceptibility.


title: Molecular characterization of HLJ-073, a recombinant canine coronavirus strain from China with an ORF3abc deletion

text: Canine enteric coronaviruses (CCoVs) are important enteric pathogens of dogs. CCoVs with different variations are typically pantropic and pathogenic in dogs. In this study, we isolated a CCoV, designated HLJ-073, from a dead 6-week-old male Pekingese with gross lesions and diarrhea. Interestingly, sequence analysis suggested that HLJ-073 contained a 350-nt deletion in ORF3abc compared with reference CCoV isolates, resulting in the loss of portions of ORF3a and ORF3c and the complete loss of ORF3b. Phylogenetic analysis based on the S gene showed that HLJ-073 was more closely related to members of the FCoV II cluster than to members of the CCoV I or CCoV II cluster. Furthermore, recombination analysis suggested that HLJ-073 originated from the recombination of FCoV 79-1683 and CCoV A76, which were both isolated in the United States. Cell tropism experiments suggested that HLJ-073 could effectively replicate in canine macrophages/monocytes and human THP-1 cells. This is the first report of the isolation of strain HLJ-073 in China, and this virus has biological characteristics that are different from those of other reported CCoVs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00705-019-04296-9) contains supplementary material, which is available to authorized users.


title: Years of life lost due to the psychosocial consequences of COVID-19 mitigation strategies based on Swiss data

text: BACKGROUND. The pandemic caused by coronavirus disease 2019 (COVID-19) has forced governments to implement strict social mitigation strategies to reduce the morbidity and mortality from acute infections. These strategies, however, carry a significant risk for mental health, which can lead to increased short-term and long-term mortality and is currently not included in modeling the impact of the pandemic. METHODS. We used years of life lost (YLL) as the main outcome measure, applied to Switzerland as an example. We focused on suicide, depression, alcohol use disorder, childhood trauma due to domestic violence, changes in marital status, and social isolation, as these are known to increase YLL in the context of imposed restriction in social contact and freedom of movement. We stipulated a minimum duration of mitigation of 3 months based on current public health plans. RESULTS. The study projects that the average person would suffer 0.205 YLL due to psychosocial consequence of COVID-19 mitigation measures. However, this loss would be entirely borne by 2.1% of the population, who will suffer an average of 9.79 YLL. CONCLUSIONS. The results presented here are likely to underestimate the true impact of the mitigation strategies on YLL. However, they highlight the need for public health models to expand their scope in order to provide better estimates of the risks and benefits of mitigation.


title: Effect of Host Species on the Distribution of Mutational Fitness Effects for an RNA Virus

text: Knowledge about the distribution of mutational fitness effects (DMFE) is essential for many evolutionary models. In recent years, the properties of the DMFE have been carefully described for some microorganisms. In most cases, however, this information has been obtained only for a single environment, and very few studies have explored the effect that environmental variation may have on the DMFE. Environmental effects are particularly relevant for the evolution of multi-host parasites and thus for the emergence of new pathogens. Here we characterize the DMFE for a collection of twenty single-nucleotide substitution mutants of Tobacco etch potyvirus (TEV) across a set of eight host environments. Five of these host species were naturally infected by TEV, all belonging to family Solanaceae, whereas the other three were partially susceptible hosts belonging to three other plant families. First, we found a significant virus genotype-by-host species interaction, which was sustained by differences in genetic variance for fitness and the pleiotropic effect of mutations among hosts. Second, we found that the DMFEs were markedly different between Solanaceae and non-Solanaceae hosts. Exposure of TEV genotypes to non-Solanaceae hosts led to a large reduction of mean viral fitness, while the variance remained constant and skewness increased towards the right tail. Within the Solanaceae hosts, the distribution contained an excess of deleterious mutations, whereas for the non-Solanaceae the fraction of beneficial mutations was significantly larger. All together, this result suggests that TEV may easily broaden its host range and improve fitness in new hosts, and that knowledge about the DMFE in the natural host does not allow for making predictions about its properties in an alternative host.


title: Retrospective comparison of convalescent plasma with continuing high-dose methylprednisolone treatment in SARS patients

text: Treatment of severe acute respiratory syndrome (SARS) is experimental, and the effectiveness of ribavirin-steroid therapy is unclear. Forty SARS patients with progressive disease after ribavirin treatment and 1.5 g of pulsed methylprednisolone were given either convalescent plasma (n = 19) or further pulsed methylprednisolone (n = 21) in a retrospective non-randomised study. Good clinical outcome was defined as discharge by day 22 following the onset of symptoms. Convalescent plasma was obtained from recovered patients after informed consent. Patients in the plasma group had a shorter hospital stay (p 0.001) and lower mortality (p 0.049) than the comparator group. No immediate adverse effects were observed following plasma infusion.


title: ERAD and ERAD tuning: disposal of cargo and of ERAD regulators from the mammalian ER

text: The endoplasmic reticulum (ER) is the site of maturation for secretory and membrane proteins in eukaryotic cells. Unsuccessful folding attempts are eventually interrupted and most folding-defective polypeptides are dislocated across the ER membrane and degraded by cytosolic proteasomes in a complex series of events collectively defined as ER-associated degradation (ERAD). Uncontrolled ERAD activity might prematurely interrupt ongoing folding programs. At steady state, this is prevented by ERAD tuning, that is, the removal of select ERAD regulators from the ER and their degradation by proteasomes and by endo-lysosomal proteases. In Coronaviruses infected cells, the formation of LC3-I coated vesicles containing ERAD regulators cleared from the ER lumen is co-opted to anchor viral replication and transcription complexes to ER-derived membranes.


title: Effective chemical virus inactivation of patient serum compatible with accurate serodiagnosis of infections

text: Abstract Objectives Highly pathogenic viruses such as EBOV are a threat to routine laboratory workers. Inactivation procedures with Triton X-100 0.1% and/or heat are currently recommended, but have unknown effects on the accuracy of serological testing. Furthermore, virus inactivation by Triton X-100 0.1% was shown to be ineffective in serum. This study aimed to demonstrate virus inactivation in serum by Triton X-100 1% and maintained accuracy of serological testing. Methods A panel of 19 serological tests was run on patient serum samples after treatment with Triton X-100 1%, 0.1%, and 0.1% + heat inactivation at 60°C for 1 h. Mean differences between measurements (bias) were calculated applying the Bland–Altman method. To determine effectiveness of virus inactivation, herpes simplex virus 1 (HSV-1) was spiked into medium containing 90% or 1% serum, and treated with Triton X-100 0.1% or 1%. Infectious titres were then determined on Vero cells. Results Serological measurements showed good agreement between controls and samples treated with Triton X-100 0.1% and 1%, with an estimated bias of 0.6 ± 9.2% (n = 258) and –0.1 ± 18.6% (n = 174), respectively. Discordant qualitative results were rare. Conversely, heat inactivation alone and combined with Triton X-100 0.1% triggered a bias of 17.5 ± 66.4% (n = 200) and 37.9 ± 79.8% (n = 160), respectively. Triton X-100 1% completely inactivated HSV-1 in 1% and 90% serum while Triton X-100 0.1% failed to do so in 90% serum. Conclusions Unlike heat inactivation, Triton X-100 1% enabled accurate serological testing and completely inactivated HSV-1 in serum. This simple method could allow safe routine serological diagnostics in high-risk patients.


Azithromycin Should Not Be Used to Treat COVID-19


title: Porcine vaccines

text: The growth of large, intensive pig operations has made effective vaccination imperative in the swine industry. This growth in size has been accompanied by outbreaks of disease such as porcine respiratory and reproductive syndrome (PRRS) and porcine epidemic diarrhea. These have caused massive losses, and effective vaccines are not yet available. Older diseases such as pleuropneumonia, erysipelas, enzootic pneumonia, proliferative enteritis, pseudorabies, and colibacillosis still persist, and classical swine fever and influenza remain as ongoing threats. The large size of many operations has required the introduction of mass vaccination procedures such as vaccination through drinking water. The current African swine fever pandemic makes the development of an effective vaccine against this virus, a matter of urgent necessity.


title: Orientation diagnostique devant une lymphopénie

text: Points essentiels Une lymphopénie est définie par un nombre de lymphocytes circulants < 1 500/mm3 chez l’adulte et 4 500/mm3 chez l’enfant avant 8 mois. Nous proposons de classer les lymphopénies en fonction de leur mécanisme supposé • les insuffisances de production, qui regroupent les déficits immunitaires primitifs ou secondaires à une dénutrition et/ou une carence en zinc • les excès de catabolisme comprenant la radiothérapie, la chimiothérapie, les traitements immunosuppresseurs, l’infection VIH et le lupus érythémateux systémique • les modifications de la répartition des lymphocytes, qui comprennent principalement l’hypersplénisme, certaines infections virales, le choc septique, les brûlures étendues, les granulomatoses et la corticothérapie • les autres causes de lymphopénie, dont le mécanisme est encore mal compris : ethnie (Éthiopiens), insuffisance rénale, lymphome, tumeur solide, lymphopénie CD4 idiopathique. Key points Lymphopenia is defined as a peripheral lymphocyte count lower than 1500/mm3 in adults and 4500/mm3 in children younger than eight months of age. We propose a classification of lymphopenia according to the mechanism involved • lymphocyte production defects, including primary immune deficiencies and immune deficiencies secondary to malnutrition or zinc deprivation • excess catabolism, due to causes including radiotherapy, chemotherapy, immunosuppressive therapy, HIV infection, and systemic lupus erythematosus • abnormal lymphocyte trapping, including mainly splenomegaly, certain viral infections, septic shock, extended burns, systemic granulomatosis, and corticosteroids • other causes of lymphocytopenia, with mechanisms that remain poorly understood: ethnicity (Ethiopians), lymphoma, renal insufficiency, and idiopathic CD4 lymphocytopenia.


The impact of COVID-19 on Acute Psychiatric Inpatient Unit


title: Human metapneumovirus in paediatric patients

text: Acute respiratory tract infections (ARTIs) are a leading cause of morbidity and mortality in children worldwide, but the aetiology of many ARTIs is still unknown. In 2001, researchers in The Netherlands reported the discovery of a previously unidentified pathogen called human metapneumovirus (hMPV). Since its initial description, hMPV has been associated with ARTI in Europe (Italy, France, Spain, the UK, Germany, Denmark, Finland and Norway), America (the USA, Canada, Argentina and Brazil), Asia (India, Japan, China and Singapore), Australia and South Africa in individuals of all ages. The incidence of infection varies from 1.5% to 25%, indicating that hMPV is a ubiquitous virus with a worldwide distribution. hMPV seems to play an important role as a cause of paediatric upper and lower respiratory tract infection, with similar, but not identical, epidemiological and clinical features to those of respiratory syncytial virus and influenza virus. Moreover, the socio-economic impact of hMPV-infected children on their families seems to be considerable, which suggests that, like influenza virus, hMPV infection may be a substantial public health problem for the community. It may be associated with significant morbidity and mortality in pre-term infants and children with underlying clinical conditions, although more adequately controlled studies are needed to confirm its importance in such patients. Many fundamental questions concerning the pathogenesis of hMPV disease and the host's specific immune response remain to be answered. Further studies are also required to properly define hMPV diagnosis, treatment and prevention strategies.