title: Acute pancreatic injuries: A complication of Stevens-Johnson syndrome/toxic epidermal necrolysis associated with cytotoxic immune cell activation

text: Abstract Background Complications involving internal organs are usually present in Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). However, pancreatic complications are rarely reported and studied. Objective To summarize clinical characteristics of SJS/TEN-associated acute pancreatic injuries (API) and to investigate underlying inflammatory mechanisms. Methods Clinical records of 124 inpatients with SJS/TEN were reviewed. Serum levels of tumor necrosis factor-α, interleukin (IL)-6, IL-18, IL-15, IL-12p70, and soluble CD56 were determined in 18 healthy donors and 17 patients with SJS/TEN, including three patients with API. Results API was diagnosed in 7.3% (9/124) of patients in the SJS/TEN cohort. Serum transaminase elevation and hypoalbuminemia occurred more frequently in patients with API compared to those without pancreatic symptoms (P = 0.004, P = 0.0002, respectively). Although API did not alter mortality rate of SJS/TEN, it was associated with longer hospitalization stays (P = 0.008). Within the serum cytokines that were elevated in SJS/TEN, only IL-18 was found to be selectively increased in patients with API compared to those without pancreatic injuries (P = 0.032). Limitations Cohort size is small. Conclusion API is a gastrointestinal complication of SJS/TEN in which hepatotoxicity is more likely to occur. Overexpression of IL-18 might be involved in this unique entity.


title: Lockdown timing and efficacy in controlling COVID-19 using mobile phone tracking

text: BACKGROUND: Italy's severe COVID-19 outbreak was addressed by a lockdown that gradually increased in space, time and intensity. The effectiveness of the lockdown has not been precisely assessed with respect to the intensity of mobility restriction and the time until the outbreak receded. METHODS: We used processed mobile phone tracking data to measure mobility restriction, and related those data to the number of new SARS-CoV-2 positive cases detected on a daily base in the three most affected Italian regions, Lombardy, Veneto and Emilia-Romagna, from February 1 through April 6, 2020, when two subsequent lockdowns with increasing intensity were implemented by the Italian government. FINDINGS: During the study period, mobility restriction was inversely related to the daily number of newly diagnosed SARS-CoV-2 positive cases only after the second, more effective lockdown, with a peak in the curve of diagnosed cases of infection occurring 14 to 18 days from lockdown in the three regions and 9 to 25 days in the included provinces. An effective reduction in transmission must have occurred nearly immediately after the tighter lockdown, given the lag time of around 10 days from asymptomatic infection to diagnosis. The period from lockdown to peak was shorter in the areas with the highest prevalence of the infection. This effect was seen within slightly more than one week in the most severely affected areas. INTERPRETATION: It appears that the less rigid lockdown led to an insufficient decrease in mobility to reverse an outbreak such as COVID-19. With a tighter lockdown, mobility decreased enough to bring down transmission promptly below the level needed to sustain the epidemic. FUNDING: No funding sources have been used for this work.


title: The UPR Branch IRE1-bZIP60 in Plants Plays an Essential Role in Viral Infection and Is Complementary to the Only UPR Pathway in Yeast

text: The unfolded protein response (UPR) signaling network encompasses two pathways in plants, one mediated by inositol-requiring protein-1 (IRE1)-bZIP60 mRNA and the other by site-1/site-2 proteases (S1P/S2P)-bZIP17/bZIP28. As the major sensor of UPR in eukaryotes, IRE1, in response to endoplasmic reticulum (ER) stress, catalyzes the unconventional splicing of HAC1 in yeast, bZIP60 in plants and XBP1 in metazoans. Recent studies suggest that IRE1p and HAC1 mRNA, the only UPR pathway found in yeast, evolves as a cognate system responsible for the robust UPR induction. However, the functional connectivity of IRE1 and its splicing target in multicellular eukaryotes as well as the degree of conservation of IRE1 downstream signaling effectors across eukaryotes remains to be established. Here, we report that IRE1 and its substrate bZIP60 function as a strictly cognate enzyme-substrate pair to control viral pathogenesis in plants. Moreover, we show that the S1P/S2P-bZIP17/bZIP28 pathway, the other known branch of UPR in plants, does not play a detectable role in virus infection, demonstrating the distinct function of the IRE1-bZIP60 pathway in plants. Furthermore, we provide evidence that bZIP60 and HAC1, products of the enzyme-substrate duet, rather than IRE1, are functionally replaceable to cope with ER stress in yeast. Taken together, we conclude that the downstream signaling of the IRE1-mediated splicing is evolutionarily conserved in yeast and plants, and that the IRE1-bZIP60 UPR pathway not only confers overlapping functions with the other UPR branch in fundamental biology but also may exert a unique role in certain biological processes such as virus-plant interactions.


title: Lactoferrin: Lactoferrin: a modulator of immune and inflammatory responses

text: Lactoferrin is an iron-binding glycoprotein of the transferrin family. Abundant expression and secretion of lactoferrin, in particular in milk and fluids of the digestive tract, are related to its implication in the first line of host defense. Lactoferrin is also a prominent component of the secondary granules of neutrophils (PMNs) and is released in infected tissues and blood during the inflammatory process. In addition to its direct antimicrobial properties, the abilities of lactoferrin to regulate the immune response and to protect against infection and septic shock have been described in numerous in vitro and in vivo studies. Although the cellular and molecular mechanisms that account for the modulation of the inflammatory and immune responses by lactoferrin are not yet totally elucidated, many are now established. At the cellular level, lactoferrin modulates the migration, maturation and function of immune cells. At the molecular level and in addition to iron binding, interactions of lactoferrin with a plethora of compounds, either soluble or membrane molecules, account for its modulatory properties. This paper reviews our current understanding of the cellular and molecular mechanisms that explain the regulatory properties of lactoferrin in host defence.


title: Severity and risk of COVID‐19 in cancer patients: An evidence based learning

text: The immune system of cancer patient gets compromised because of cancer therapy, surgery and malignancy and thus the probability of infection are increased than the general patients. Immunosuppression can expose cancer patients to serious complications which can lead to delay in diagnosis and unnecessary hospitalizations that may adversely affect the prognosis of the disease. Patients who received chemotherapy or surgery within the 30 days before COVID‐19 pandemic have more risk of infection than the patients who had not undergone chemotherapy or surgery. This article is protected by copyright. All rights reserved.


title: Phylogenetic analysis of avian infectious bronchitis virus strains isolated in Japan

text: To define the origin and evolution of recent avian infectious bronchitis virus (IBV) in Japan, a genetic analysis was performed. By phylogenetic analysis based on the S1 gene including the sequence of the hypervariable regions, IBV isolates in Japan were classified into five genetic groups, which included two already-known groups (Mass and Gray). Among them, three major genetic groups were associated with the recent outbreaks of IB in Japan. One group is indigenous to Japan and could not be placed within the known existing groups in other countries. The remaining two groups, which have emerged recently, are related to isolates in China and Taiwan.


title: Influence of hydrophobic and electrostatic residues on SARS-coronavirus S2 protein stability: Insights into mechanisms of general viral fusion and inhibitor design

text: Severe acute respiratory syndrome (SARS) is an acute respiratory disease caused by the SARS-coronavirus (SARS-CoV). SARS-CoV entry is facilitated by the spike protein (S), which consists of an N-terminal domain (S1) responsible for cellular attachment and a C-terminal domain (S2) that mediates viral and host cell membrane fusion. The SARS-CoV S2 is a potential drug target, as peptidomimetics against S2 act as potent fusion inhibitors. In this study, site-directed mutagenesis and thermal stability experiments on electrostatic, hydrophobic, and polar residues to dissect their roles in stabilizing the S2 postfusion conformation was performed. It was shown that unlike the pH-independent retroviral fusion proteins, SARS-CoV S2 is stable over a wide pH range, supporting its ability to fuse at both the plasma membrane and endosome. A comprehensive SARS-CoV S2 analysis showed that specific hydrophobic positions at the C-terminal end of the HR2, rather than electrostatics are critical for fusion protein stabilization. Disruption of the conserved C-terminal hydrophobic residues destabilized the fusion core and reduced the melting temperature by 30°C. The importance of the C-terminal hydrophobic residues led us to identify a 42-residue substructure on the central core that is structurally conserved in all existing CoV S2 fusion proteins (root mean squared deviation = 0.4 Å). This is the first study to identify such a conserved substructure and likely represents a common foundation to facilitate viral fusion. We have discussed the role of key residues in the design of fusion inhibitors and the potential of the substructure as a general target for the development of novel therapeutics against CoV infections.


title: Expression and purification of an influenza hemagglutinin—one step closer to a recombinant protein-based influenza vaccine

text: Numerous human infections with avian influenza viruses in Asia in recent years have raised the concern that the next influenza pandemic is imminent. The most effective way to combat influenza is through the vaccination of the public. However, a minimum of 3–6 months is needed to develop an influenza vaccine using the traditional egg-based vaccine approach. The influenza hemagglutinin protein (HA), the active ingredient in the current vaccine, can be expressed in insect cells using the baculovirus expression vector system and purified rapidly. An influenza vaccine based on such a recombinant antigen allows a more timely response to a potential influenza pandemic. Here, we report an innovative monitoring assay for recombinant HA (rHA) expression and a rapid purification process. Various biochemical analyses indicate that the purified rHA is properly folded and biologically active.


The eighth alternative to Evidence Based Medicine in the early era of the COVID-19 pandemic: too much Emergency and Emotion, too little Evidence.


title: Identification of a Novel Betacoronavirus (Merbecovirus) in Amur Hedgehogs from China

text: While dromedaries are the immediate animal source of Middle East Respiratory Syndrome (MERS) epidemic, viruses related to MERS coronavirus (MERS-CoV) have also been found in bats as well as hedgehogs. To elucidate the evolution of MERS-CoV-related viruses and their interspecies transmission pathway, samples were collected from different mammals in China. A novel coronavirus related to MERS-CoV, Erinaceus amurensis hedgehog coronavirus HKU31 (Ea-HedCoV HKU31), was identified from two Amur hedgehogs. Genome analysis supported that Ea-HedCoV HKU31 represents a novel species under Merbecovirus, being most closely related to Erinaceus CoV from European hedgehogs in Germany, with 79.6% genome sequence identity. Compared to other members of Merbecovirus, Ea-HedCoV HKU31 possessed unique non-structural proteins and putative cleavage sites at ORF1ab. Phylogenetic analysis showed that Ea-HedCoV HKU31 and BetaCoV Erinaceus/VMC/DEU/2012 were closely related to NeoCoV and BatCoV PREDICT from African bats in the spike region, suggesting that the latter bat viruses have arisen from recombination between CoVs from hedgehogs and bats. The predicted HKU31 receptor-binding domain (RBD) possessed only one out of 12 critical amino acid residues for binding to human dipeptidyl peptidase 4 (hDPP4), the MERS-CoV receptor. The structural modeling of the HKU31-RBD-hDPP4 binding interphase compared to that of MERS-CoV and Tylonycteris bat CoV HKU4 (Ty-BatCoV HKU4) suggested that HKU31-RBD is unlikely to bind to hDPP4. Our findings support that hedgehogs are an important reservoir of Merbecovirus, with evidence of recombination with viruses from bats. Further investigations in bats, hedgehogs and related animals are warranted to understand the evolution of MERS-CoV-related viruses.


El CIBERESP durante la pandemia de COVID-19


title: Challenges in Dengue Vaccines Development: Pre-existing Infections and Cross-Reactivity

text: Dengue is one of the most frequently transmitted mosquito-borne diseases in the world, which creates a significant public health concern globally, especially in tropical and subtropical countries. It is estimated that more than 390 million people are infected with dengue virus each year and around 96 million develop clinical pathologies. Dengue infections are not only a health problem but also a substantial economic burden. To date, there are no effective antiviral therapies and there is only one licensed dengue vaccine that only demonstrated protection in the seropositive (Immune), naturally infected with dengue, but not dengue seronegative (Naïve) vaccines. In this review, we address several immune components and their interplay with the dengue virus. Additionally, we summarize the literature pertaining to current dengue vaccine development and advances. Moreover, we review some of the factors affecting vaccine responses, such as the pre-vaccination environment, and provide an overview of the significant challenges that face the development of an efficient/protective dengue vaccine including the presence of multiple serotypes, antibody-dependent enhancement (ADE), as well as cross-reactivity with other flaviviruses. Finally, we discuss targeting T follicular helper cells (Tfh), a significant cell population that is essential for the production of high-affinity antibodies, which might be one of the elements needed to be specifically targeted to enhance vaccine precision to dengue regardless of dengue serostatus.


Síndrome de DRESS simulando enfermedad por coronavirus 2019-NcoV


title: Implications of COVID-19 Outbreak on Immune Therapies in Multiple Sclerosis Patients—Lessons Learned From SARS and MERS

text: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic keeps the world in suspense. In addition to the fundamental challenges for the health care system, the individual departments must decide how to deal with patients at risk. Neurologists are confronted with the question, how they should advise their patients regarding immunosuppressive treatment. In particular, the large number of different disease-modifying therapies (DMTs) in the treatment of neuroimmunological diseases such as multiple sclerosis poses a challenge. To a limited extent, it might be useful to transfer knowledge from previous SARS- and Middle East respiratory syndrome (MERS) coronavirus outbreaks in 2002/2003 and 2012 to the current situation. Overall, immunosuppressive therapy does neither seem to have a major impact on infection with SARS- and MERS-CoV nor does it seem to lead to a severe disease course in many cases. Considering the immunological responses against infections with novel coronaviruses in humans, interferons, glatiramer acetate, and teriflunomide appear to be safe. As lymphopenia seems to be associated with a more severe disease course, all DMTs causing lymphopenia, such as cladribine, alemtuzumab, and dimethyl fumarate, need to be reviewed more thoroughly. As they are, in general, associated with a higher risk of infection, depleting anti-CD20 antibodies may be problematic drugs. However, it has to be differentiated between the depletion phase and the phase of immune reconstitution. In summary, previous coronavirus outbreaks have not shown an increased risk for immunocompromised patients. Patients with severe neuroimmunological diseases should be kept from hasty discontinuation of immunotherapy.


title: Cardiac Involvment in COVID-19–Related Acute Respiratory Distress Syndrome

text: The cardiac involvement in (COVID-19) Coronavirus disease is still under evaluation, especially in severe COVID-19-related Acute Respiratory Distress Syndrome (ARDS). The cardiac involvement was assessed by serial troponin levels and echocardiograms in 28 consecutive patients with COVID-19 ARDS consecutively admitted to our Intensive Care Unit from 1(st) March to 31(th) March. Twenty-eight COVID-19 patients (aged 61.7 ± 10 years, males 79%). The majority was mechanically ventilated (86%) and four patients (14%) required veno-venous extracorporeal membrane oxygenation (ECMO). As of 31(th) March, the ICU mortality rate was 7%, while 7 patients was discharged (25%) with a length of stay of 8.2 ±5 days. At echocardiographic assessment on admission, acute core pulmonale was detected in two patients who required ECMO support. Increased systolic arterial pressure was detected in all patients. Increased Troponin T levels were detectable in 11 patients (39%) on admission. At linear regression analysis, troponin T showed a direct relationship with CRP (R square: 0.082, F: 5.95, p=0.017). In conclusions, in COVID-19-related ARDS, increased in Tn levels was common but not associated with alterations in wall motion kinesis, thus suggesting that troponin T elevation is likely to be multifactorial, mainly linked to disease severely (as inferred by the relation between Tn and CRP). The increase in systolic pulmonary arterial pressures observed in all patients may be related to hypoxic vasoconstriction. Further studies are need to confirm our findings in larger cohorts.


title: SARS‐CoV‐2 infection anxieties and general population restrictions delay diagnosis and treatment of acute haematological malignancies

text: Governments worldwide have responded to the SARS-CoV-2 pandemic by officially imposing or unofficially encouraging social distancing measures. These measures have had a positive impact on the spread of the virus, however the "stay at home" formula and the lack of specific scientific information about SARS-CoV-2 has understandably resulted in increased anxiety and fear. One unforeseen consequence has been an enhanced risk of underestimating the symptoms and clinical signs of others pathologies even more severe than the viral infection. In the last 35 days, we have observed 8 patients who delayed coming to our hospital emergency room despite unmistakable clinical symptoms and signs of acute haematological disease, putting themselves at risk of compromising their entire therapeutic plan. The similarities between viral symptoms and initial clinical signs of haematological pathologies and the anxiety of contracting SARS-CoV-2 infection were the main causes of this delay.


title: The DE and FG loops of the HPV major capsid protein contribute to the epitopes of vaccine-induced cross-neutralising antibodies

text: The human papillomavirus (HPV) vaccines consist of major capsid protein (L1) virus-like particles (VLP) and are highly efficacious against the development of cervical cancer precursors attributable to oncogenic genotypes, HPV16 and HPV18. A degree of vaccine-induced cross-protection has also been demonstrated against genetically-related genotypes in the Alpha-7 (HPV18-like) and Alpha-9 (HPV16-like) species groups which is coincident with the detection of L1 cross-neutralising antibodies. In this study the L1 domains recognised by inter-genotype cross-neutralising antibodies were delineated. L1 crystallographic homology models predicted a degree of structural diversity between the L1 loops of HPV16 and the non-vaccine Alpha-9 genotypes. These structural predictions informed the design of chimeric pseudovirions with inter-genotype loop swaps which demonstrated that the L1 domains recognised by inter-genotype cross-neutralising antibodies comprise residues within the DE loop and the late region of the FG loop. These data contribute to our understanding of the L1 domains recognised by vaccine-induced cross-neutralising antibodies. Such specificities may play a critical role in vaccine-induced cross-protection.


title: Medidas de contención de la violencia de género durante la pandemia de COVID-19

text: Resumen Las crisis, las emergencias y las épocas de disturbios se han relacionado con un aumento de la violencia interpersonal, incluida la violencia hacia las mujeres. A raíz de la declaración de estado de alarma y confinamiento se han puesto en marcha nuevas medidas encaminadas a paliar el posible impacto de la violencia de género (Plan de contingencia contra la violencia de género ante la crisis del coronavirus o Real Decreto-Ley de medidas urgentes). Se revisan en este documento las medidas de contención adoptadas hasta el momento por el gobierno de España y las comunidades autónomas, y las iniciativas formuladas en diferentes países. Ante la ausencia de medidas económicas concretas planteadas hasta la fecha y el escenario de incertidumbre económica concluimos que no es posible prevenir la violencia de género de manera integral sin considerar el aumento del desempleo, la temporalidad y la inestabilidad laboral, la dependencia económica o la sobrecarga de tareas reproductivas, entre otros elementos que la facilitan. Abstract Crises, emergencies and times of unrest have been linked to increased interpersonal violence, including violence against women. Following the declaration of alarm status and quarantine, different measures have been implemented to mitigate the possible effect of gender violence (Contingency Plan against Gender-Based Violence in Coronavirus Crisis or Royal Decree Law on Emergency Measures). This document reviews the measures adopted so far by the government of Spain, the autonomous governments and the initiatives formulated in different countries. In the absence of concrete economic measures to date, and the scenario of economic uncertainty, we conclude that it is not possible to prevent gender-based violence in a comprehensive way, without considering the increase in unemployment, temporary and instability employment, economic dependency or the overload of household chores and reproductive tasks, among other elements that facilitate it.


title: Framework for prioritizing head and neck surgery during the COVID‐19 pandemic

text: The COVID‐19 pandemic has placed an extraordinary demand on the United States health care system. Many institutions have canceled elective and non‐urgent procedures to conserve resources and limit exposure. While operational definitions of elective and urgent categories exist, there is a degree of surgeon judgment in designation. In the present commentary, we provide a framework for prioritizing head and neck surgery during the pandemic. Unique considerations for the head and neck patient are examined including risk to the oncology patient, outcomes following delay in head and neck cancer therapy, and risk of transmission during otolaryngologic surgery. Our case prioritization criteria consist of four categories: urgent—proceed with surgery, less urgent—consider postpone > 30 days, less urgent—consider postpone 30 to 90 days, and case‐by‐case basis. Finally, we discuss our preoperative clinical pathway for transmission mitigation including defining low‐risk and high‐risk surgery for transmission and role of preoperative COVID‐19 testing.


title: Associations between exposure to viruses and bovine respiratory disease in Australian feedlot cattle

text: Abstract Bovine respiratory disease (BRD) is the most important cause of clinical disease and death in feedlot cattle. Respiratory viral infections are key components in predisposing cattle to the development of this disease. To quantify the contribution of four viruses commonly associated with BRD, a case-control study was conducted nested within the National Bovine Respiratory Disease Initiative project population in Australian feedlot cattle. Effects of exposure to Bovine viral diarrhoea virus 1 (BVDV-1), Bovine herpesvirus 1 (BoHV-1), Bovine respiratory syncytial virus (BRSV) and Bovine parainfluenza virus 3 (BPIV-3), and to combinations of these viruses, were investigated. Based on weighted seroprevalences at induction (when animals were enrolled and initial samples collected), the percentages of the project population estimated to be seropositive were 24% for BoHV-1, 69% for BVDV-1, 89% for BRSV and 91% for BPIV-3. For each of the four viruses, seropositivity at induction was associated with reduced risk of BRD (OR: 0.6–0.9), and seroincrease from induction to second blood sampling (35–60 days after induction) was associated with increased risk of BRD (OR: 1.3–1.5). Compared to animals that were seropositive for all four viruses at induction, animals were at progressively increased risk with increasing number of viruses for which they were seronegative; those seronegative for all four viruses were at greatest risk (OR: 2.4). Animals that seroincreased for one or more viruses from induction to second blood sampling were at increased risk (OR: 1.4–2.1) of BRD compared to animals that did not seroincrease for any viruses. Collectively these results confirm that prior exposure to these viruses is protective while exposure at or after feedlot entry increases the risk of development of BRD in feedlots. However, the modest increases in risk associated with seroincrease for each virus separately, and the progressive increases in risk with multiple viral exposures highlights the importance of concurrent infections in the aetiology of the BRD complex. These findings indicate that, while efficacious vaccines could aid in the control of BRD, vaccination against one of these viruses would not have large effects on population BRD incidence but vaccination against multiple viruses would be expected to result in greater reductions in incidence. The findings also confirm the multifactorial nature of BRD development, and indicate that multifaceted approaches in addition to efficacious vaccines against viruses will be required for substantial reductions in BRD incidence.


title: Acute bronchitis: a review of diagnosis and evidence-based management

text: Abstract Obstetricians and gynecologists are increasingly involved in primary care. Acute bronchitis is among the most common ambulatory complaints. Although the cause of acute bronchitis is predominantly viral, 50–70% of patients presenting with this condition are treated with antibiotics. Because of the increasing bacterial resistance to antibiotics, the cost of prescription drugs, and the potential adverse reactions to them, the present management of acute bronchitis has important shortcomings. Also, inhaled bronchodilators are underused for symptomatic management. Improved awareness among physicians about the recommended management of acute bronchitis has been targeted as an important means of decreasing unnecessary antibiotic use. Patient satisfaction motivates physicians to prescribe antibiotics in managing acute bronchitis. However, patient satisfaction does not necessarily correlate with prescribing of antibiotics but rather with patient education. We present a review of the diagnosis and differential diagnosis of acute bronchitis and its management.


title: Glycyrrhizic acid as the antiviral component of Glycyrrhiza uralensis Fisch. against coxsackievirus A16 and enterovirus 71 of hand foot and mouth disease

text: Abstract Ethnopharmacological relevance The radices of Glycyrrhiza uralensis Fisch. and herbal preparations containing Glycyrrhiza spp. have been used for thousands of years as an herbal medicine for the treatment of viral induced cough, viral hepatitis, and viral skin diseases like ulcers in China. Glycyrrhizic acid (GA) is considered the principal component in Glycyrrhiza spp. with a wide spectrum of antiviral activity. Aim The present study attempt to validate the medicinal use of Glycyrrhiza uralensis for hand, foot and mouth disease (HFMD) and further to verify whether GA is an active antiviral component in the water extract of Glycyrrhiza uralensis. Materials and methods Radices of Glycyrrhiza uralensis Fisch. were extracted with hot water. The chemical contents of the extract were profiled with HPLC analysis. The antiviral activity of the extract and the major components was evaluated against infection of enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) on Vero cells. The cytopathic effect caused by the infection was measured with MTT assay. Infectious virion production was determined using secondary infection assays and viral protein expression by immunoblotting analysis. Results The extract at 1000μg/ml suppressed EV71 replication by 1.0 log and CVA16 by 1.5 logs. The antiviral activity was associated with the content of GA in the extract since selective depletion of GA from the extract by acid precipitation resulted in loss of antiviral activity. In contrast, the acid precipitant retained antiviral activity. The precipitant at a concentration of 200μg/ml inhibited EV71 and CVA16 replication by 1.7 and 2.2 logs, respectively. Furthermore, GA dose-dependently blocked viral replication of EV71 and CVA16. At 3mM, GA reduced infectious CVA16 and EV71 production by 3.5 and 2.2 logs, respectively. At 5mM, CVA16 production was reduced by 6.0 logs and EV71 by 4.0 logs. Both EV71 and CVA16 are members of Enterovirus genus, time-of-drug addition studies however showed that GA directly inactivated CVA16, while GA anti-EV71 effect was associated with an event(s) post virus cell entry. Conclusions This study validated the medicinal usefulness of radices Glycyrrhiza uralensis against the etiological agents of HFMD. In addition to the identification of GA as the antiviral component of Glycyrrhiza uralensis against EV71 and CVA16 infection, this study also reveals that GA inhibits EV71 and CVA16 with distinct mechanisms.


title: A commentary on the COVID‐19 crisis, sustainability and the service industries

text: Over the last two decades sustainability strategies and programmes have assumed an increasingly high profile within the leading companies in the service industries. However, the Covid‐19 (Coronavirus) poses a major challenge to the majority of companies within these industries and may reduce their commitment to sustainable development. At the same time, a counter argument holds that a continuing commitment to sustainability will be vitally important in maintaining the relationships between the service industries and the natural and social capital on which many of them depend. This commentary paper explores sustainability within the retail and hospitality industries through the lens of the COVID‐19 crisis. The paper provides an outline of the COVID‐19 crisis and of its impact within the retail and hospitality industries and explores some of the relationships between COVID‐19 and sustainability.


title: 8 Animal biosafety

text: Abstract Aside from playing a vital part in overseeing the use of recombinant or synthetic nucleic acids used in vitro, in vivo, in utero, or ex vivo, Institutional Biosafety Committees (IBCs) oversee multiple aspects of research ranging from: performing risk assessments and mitigation, including assignment of biosafety levels and practices, NIH category designations related to the particular experiments per the NIH guidelines for research involving synthetic or nucleic acid molecules, agency reporting and approval of standard operation procedures used for animal research. Animal models are a core component of biological research and play a critical role in numerous aspects of studying human disease, including through the creation of transgenic animals, but they can also be used to research diseases that naturally occur within animal populations. They also present disadvantages and unique biosafety issues.


title: Chlamydia trachomatis, Bordetella pertussis and other respiratory bacteria in the aetiology of lower respiratory tract infections in young infants

text: Systematic testing for Bordetella pertussis in young infants with lower respiratory tract infections (LRTIs) has been suggested, because the early clinical characteristics are difficult to recognise and distinguish from viral infections (1). Chlamydia trachomatis, a sexually transmitted disease passed on during birth, is also important (2-3). Recently, multiplex polymerase chain reaction (PCR) panels that include other respiratory bacteria have been implemented in clinical practice. Several studies have indicated that the atypical pathogens Mycoplasma pneumoniae and Chlamydia pneumoniae can cause community-acquired LRTIs in older children (4-5), but their role in young infants remains largely unknown. This article is protected by copyright. All rights reserved.


title: Prevalence of venous thromboembolism in patients with severe novel coronavirus pneumonia

text: BACKGROUND: Three months ago, severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) broke out in Wuhan, China, and spread rapidly around the world. Severe novel coronavirus pneumonia (NCP) patients have abnormal blood coagulation function, but their venous thromboembolism (VTE) prevalence is still rarely mentioned. OBJECTIVES: To determine the incidence of VTE in patients with severe NCP. METHODS: In this study, 81 severe NCP patients in the intensive care unit (ICU) of Union Hospital (Wuhan, China) were enrolled. The results of conventional coagulation parameters and lower limb vein ultrasonography of these patients were retrospectively collected and analyzed. RESULTS: The incidence of VTE in these patients was 25% (20/81), of which 8 patients with VTE events died. The VTE group was different from the non‐VTE group in age, lymphocyte counts, activated partial thromboplastin time (APTT), D‐dimer, etc. If 1.5 µg/mL was used as the D‐dimer cut‐off value to predicting VTE, the sensitivity was 85.0%, the specificity was 88.5%, and the negative predictive value (NPV) was 94.7%. CONCLUSIONS: The incidence of VTE in patients with severe NCP is 25% (20/81), which may be related to poor prognosis. The significant increase of D‐dimer in severe NCP patients is a good index for identifying high‐risk groups of VTE.


title: Respiratory virus-induced heterologous immunity: Part of the problem or part of the solution?

text: PURPOSE: To provide current knowledge on respiratory virus-induced heterologous immunity (HI) with a focus on humoral and cellular cross-reactivity. Adaptive heterologous immune responses have broad implications on infection, autoimmunity, allergy and transplant immunology. A better understanding of the mechanisms involved might ultimately open up possibilities for disease prevention, for example by vaccination. METHODS: A structured literature search was performed using Medline and PubMed to provide an overview of the current knowledge on respiratory-virus induced adaptive HI. RESULTS: In HI the immune response towards one antigen results in an alteration of the immune response towards a second antigen. We provide an overview of respiratory virus-induced HI, including viruses such as respiratory syncytial virus (RSV), rhinovirus (RV), coronavirus (CoV) and influenza virus (IV). We discuss T cell receptor (TCR) and humoral cross-reactivity as mechanisms of HI involving those respiratory viruses. Topics covered include HI between respiratory viruses as well as between respiratory viruses and other pathogens. Newly developed vaccines, which have the potential to provide protection against multiple virus strains are also discussed. Furthermore, respiratory viruses have been implicated in the development of autoimmune diseases, such as narcolepsy, Guillain-Barré syndrome, type 1 diabetes or myocarditis. Finally, we discuss the role of respiratory viruses in asthma and the hygiene hypothesis, and review our recent findings on HI between IV and allergens, which leads to protection from experimental asthma. CONCLUSION: Respiratory-virus induced HI may have protective but also detrimental effects on the host. Respiratory viral infections contribute to asthma or autoimmune disease development, but on the other hand, a lack of microbial encounter is associated with an increasing number of allergic as well as autoimmune diseases. Future research might help identify the elements which determine a protective or detrimental outcome in HI-based mechanisms.


title: The microbiota of the respiratory tract: gatekeeper to respiratory health

text: The respiratory tract is a complex organ system that is responsible for the exchange of oxygen and carbon dioxide. The human respiratory tract spans from the nostrils to the lung alveoli and is inhabited by niche-specific communities of bacteria. The microbiota of the respiratory tract probably acts as a gatekeeper that provides resistance to colonization by respiratory pathogens. The respiratory microbiota might also be involved in the maturation and maintenance of homeostasis of respiratory physiology and immunity. The ecological and environmental factors that direct the development of microbial communities in the respiratory tract and how these communities affect respiratory health are the focus of current research. Concurrently, the functions of the microbiome of the upper and lower respiratory tract in the physiology of the human host are being studied in detail. In this Review, we will discuss the epidemiological, biological and functional evidence that support the physiological role of the respiratory microbiota in the maintenance of human health. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nrmicro.2017.14) contains supplementary material, which is available to authorized users.


title: Development of a modified immunoelectroosmophoresis method for Uukuniemi and Runde virus serology

text: In search for a suitable method for sero-ecological screenings for arboviruses in Norway, efforts were undertaken to make the immunoelectroosmophoresis technique more sensitive than here to fare in detection of antibodies. The aim was to make it comparable to haemagglutination inhibition test in sensitivity, retaining the advantages in specificity, simplicity and capacity. This has been achieved by: 1. Using concentrated virus as antigen. 2. Performing electrophoresis in a gel consisting of an agar-agarose mixture in optimal concentrations. 3. “Sandwiching” the first specific electrophoretic run with an anti-species antiserum to the tested sample.


title: Anti-tumor effects of Abnormal Savda Munziq on the transplanted cervical cancer (U27) mouse model

text: BACKGROUND: Abnormal Savda Munziq (ASMq), a traditional uyghur medicine, has shown anti-tumour properties in vitro. it was showed that total flavonoids of ASMq could inhibit the proliferation and enhance the antioxidant ability of human cervix cancer HeLa cell. This study attempts to confirm these effects on the transplanted cervical cancer (U27) mouse model in vivo. METHODS: Forty eight Kunming mice were randomly divided in to six groups: normal control group (Control group), U27 tumor model group (Model group), cyclophosphamide administration group (CTX group),low-dose ASMq group (ASMq.L group), medium-dose ASMq group (ASMq.M group), and high-dose ASMq group (ASMq.H group). The five groups except normal control group transplanted with cervical cancer (U27) cells. We observed mice tumor inhibition rate and conducted the histopathological analysisUsing the western blot assay, the expression of TGF-β1 and TNF-α protein in transplanted cervical cancer U27 tumor tissue were detected. RESULTS: The tumor inhibition rates of CTX group, ASMq.L group, ASMq.M group, and ASMq.H group were 72.21, 31.27, 60.53 and 51.94% respectively, has obvious antitumor effect. ASMq significantly promote the spleen tlymphocyte proliferation of transplanted cervical cancer U27 mice. Invasive growth and diffusion rate in tumor tissue were accelerate in the transplanted cervical cancer U27 model group. Tumor tissue necrosis of tumor cells are smaller in the medium, high dosage group. Compared with the U27 model group, the expression levels of TGF-β1 protein and TNF-α protein expression exhibited statistically significant decreased in the mice tumor tissues in the CTX administration group and the ASMq administration group. CONCLUSIONS: ASMq has some antitumor effects on U27 model mice in vivo, The effects are achieved not only by improving the immune function of U27 model mice, but also by inhibiting the expression levels of TGF-β1 protein while promoting the expression levels of TNF-α protein.


title: Considerations for head and neck oncology practices during the coronavirus disease 2019 (COVID‐19) pandemic: Wuhan and Toronto experience

text: The practices of head and neck surgical oncologists must evolve to meet the unprecedented needs placed on our health care system by the Coronavirus disease 2019 (COVID‐19) pandemic. Guidelines are emerging to help guide the provision of head and neck cancer care, though in practice, it can be challenging to operationalize such recommendations. Head and neck surgeons at Wuhan University faced significant challenges in providing care for their patients. Similar challenges were faced by the University of Toronto during the severe acute respiratory syndrome (SARS) pandemic in 2003. Herein, we outline our combined experience and key practical considerations for maintaining an oncology service in the midst of a pandemic.


title: Dry heat and microwave generated steam protocols for the rapid decontamination of respiratory personal protective equipment in response to COVID-19-related shortages

text: BACKGROUND: In the wake of SARS-CoV-2 pandemic and unprecedented global demand, clinicians are struggling to source adequate access to personal protective equipment. Respirators can be in short supply, though are necessary to protect workers from SARS-CoV-2 exposure. Rapid decontamination and reuse of respirators may provide relief for the strained procurement situation. METHOD: In this study, we investigated the suitability of 70(o)C dry heat and microwave-generated steam (MGS) for reprocessing of FFP2/N95-type respirators, and Type-II surgical face masks. S. aureus was used as surrogate as it is less susceptible than enveloped viruses to chemical and physical processes. RESULTS: We observed >4 log(10) reductions in the viability of dry S. aureus treated by dry heat for 90 minutes at 70°C and >6 log(10) reductions by MGS in 90 seconds. After 3 reprocessing cycles, neither process was found to negatively impact the bacterial or NaCl filtration efficiency of the respirators that were tested. However, MGS was incompatible with Type-II surgical masks tested, as we confirmed that bacterial filtration capacity was completely lost following reprocessing. MGS was observed to be incompatible with some respirator types due to arcing observed around some types of metal nose clips and by loss of adhesion of clips to the mask. CONCLUSION: Considering the advantages and disadvantages of each approach, we propose a reprocessing PPE/face mask workflow for use in medical areas.


Crash Course in Decision Making


title: COVID-19 presenting as severe, persistent abdominal pain and causing late respiratory compromise in a 33-year-old man

text: A 33-year-old man presented repeatedly with severe abdominal pain and diarrhoea. Renal colic was suspected, and he was admitted for pain management. Questioning elicited an additional history of sore throat and mild, dry cough. Inflammatory markers were mildly raised (C reactive protein (CRP) 40 mg/L). Initial nasopharyngeal swabs were negative for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) by PCR. CT of the kidneys, ureters and bladder (CT KUB) was normal; however, CT of the thorax showed multifocal bilateral peripheral areas of consolidation consistent with COVID-19 infection. He developed respiratory compromise and was transferred to the intensive care unit (ICU). Sputum was positive for SARS-CoV-2 by PCR, and culture grew Yersinia enterocolitica. He recovered following supportive management and treatment with piperacillin–tazobactam.


title: Some properties of a simple stochastic epidemic model of SIR type

text: Abstract We investigate the properties of a simple discrete time stochastic epidemic model. The model is Markovian of the SIR type in which the total population is constant and individuals meet a random number of other individuals at each time step. Individuals remain infectious for R time units, after which they become removed or immune. Individual transition probabilities from susceptible to diseased states are given in terms of the binomial distribution. An expression is given for the probability that any individuals beyond those initially infected become diseased. In the model with a finite recovery time R, simulations reveal large variability in both the total number of infected individuals and in the total duration of the epidemic, even when the variability in number of contacts per day is small. In the case of no recovery, R =∞, a formal diffusion approximation is obtained for the number infected. The mean for the diffusion process can be approximated by a logistic which is more accurate for larger contact rates or faster developing epidemics. For finite R we then proceed mainly by simulation and investigate in the mean the effects of varying the parameters p (the probability of transmission), R, and the number of contacts per day per individual. A scale invariant property is noted for the size of an outbreak in relation to the total population size. Most notable are the existence of maxima in the duration of an epidemic as a function of R and the extremely large differences in the sizes of outbreaks which can occur for small changes in R. These findings have practical applications in controlling the size and duration of epidemics and hence reducing their human and economic costs.


title: Health evaluation of African penguins (Spheniscus demersus) in southern Africa

text: The African penguin (Spheniscus demersus) is an endangered seabird that breeds along the coast of Namibia and South Africa, and disease surveillance was identified as a priority for its conservation. Aiming for the establishment of baseline data on the presence of potential pathogens in this species, a comprehensive health assessment (blood smear examination, haematology, biochemistry and serology) was conducted on samples obtained from 578 African penguins at 11 breeding colonies and a rehabilitation centre. There were 68 penguins that were seropositive for at least one of seven pathogens tested: avian encephalomyelitis virus, avian infectious bronchitis virus, avian reovirus, infectious bursal disease virus, Newcastle disease virus, Mycoplasma gallisepticum and Mycoplasma synoviae. All samples were seronegative for avian influenza virus subtypes H5 and H7 and infectious laryngotracheitis virus. The apparent prevalence of Babesia sp. and Borrelia sp. in blood smears was consistent with previous studies. Babesia-infected individuals had a regenerative response of the erythrocytic lineage, an active inflammatory response and hepatic function impairment. These findings indicate that African penguins may be exposed to conservation-significant pathogens in the wild and encourage further studies aiming for the direct detection and/or isolation of these microorganisms.


title: Structure of aminopeptidase N from Escherichia coli complexed with the transition‐state analogue aminophosphinic inhibitor PL250

text: Aminopeptidase N (APN; EC 3.4.11.2) purified from Escherichia coli has been crystallized with the optically pure aminophosphinic inhibitor PL250, H(3)N(+)‐CH(CH(3))‐P(O)(OH)‐CH(2)‐CH(CH(2)Ph)‐CONH‐CH(CH(2)Ph)CO(2) (−), which mimics the transition state of the hydrolysis reaction. PL250 inhibits APN with a K (i) of 1.5–2.2 nM and its three‐dimensional structure in complex with E. coli APN showed its interaction with the S(1), S′(1) and S′(2) subsites of the catalytic site. In this structure, the Zn ion was shown to be pentacoordinated by His297, His301 and Glu320 of APN and the two O atoms of the phosphinic moiety of PL250. One of these O atoms is also involved in a hydrogen bond to Tyr381, supporting the proposed role of this amino acid in the stabilization of the transition state of the enzymatic process. The strength of the phosphinic zinc binding and the occupancy of the S′(2) subsite account for the 100‐fold increase in affinity of PL250 compared with the dipeptide‐derived inhibitor bestatin (K (i) = 4.1 × 10(−6) M). Accordingly, the removal of the C‐terminal phenylalanine of PL250 resulted in a large decrease in affinity (K (i) = 2.17 × 10(−7) M). Furthermore, it was observed that the C‐terminal carboxyl group of the inhibitor makes no direct interactions with the amino acids of the APN active site. Interestingly, PL250 exhibits the same inhibitory potency for E. coli APN and for mammalian enzymes, suggesting that the structure of the complex could be used as a template for the rational design of various human APN inhibitors needed to study the role of this aminopeptidase in various pathologies.


title: Ojo y piel

text: Las relaciones entre la patología del ojo y de su revestimiento cutáneo son muy numerosas. Debido a las analogías embriológicas e histológicas, sus principales consecuencias oftalmológicas afectan a la superficie ocular en el sentido más amplio: tegumentos de los párpados, conjuntivas y córnea. La exploración clínica dermatológica sospechará estas alteraciones, que confirmará el oftalmólogo. No obstante, algunos trastornos pueden afectar a todos los demás tejidos oculares. Ya se trate de una localización ocular de algunas enfermedades dermatológicas, como las dermatosis infecciosas, o de auténticos síndromes oculocutáneos, la exploración clínica especializada, y a veces repetida, suele orientar el tratamiento. Las consecuencias sobre la función visual de algunas de estas alteraciones pueden ser graves. La epidermólisis tóxica de Lyell constituye el ejemplo extremo que puede provocar una ceguera corneal bilateral de tratamiento temible. La dermatitis atópica es otro ejemplo de enfermedad más frecuente, pero cuyas complicaciones oculares también pueden amenazar la visión si el tratamiento oftalmodermatológico no es correcto. El ojo y la piel se asocian de nuevo en numerosas enfermedades generales, congénitas, hereditarias o no, o bien adquiridas, cuyos síntomas específicos relacionados con ambas especialidades ayudan a establecer el diagnóstico positivo. Los ejemplos más frecuentes son el lupus, la enfermedad de Behçet, la sarcoidosis y las facomatosis. La superficie ocular conjuntivocorneal comparte con la piel una cierta similitud en cuanto al desarrollo de tumores, pigmentados o no. Por último, merecen describirse las consecuencias oculares de algunos tratamientos dermatológicos. También se hará mención a las situaciones inversas, que son más excepcionales.


title: Accurate Reproduction of 161 Small-Molecule Complex Crystal Structures using the EUDOC Program: Expanding the Use of EUDOC to Supramolecular Chemistry

text: EUDOC is a docking program that has successfully predicted small-molecule-bound protein complexes and identified drug leads from chemical databases. To expand the application of the EUDOC program to supramolecular chemistry, we tested its ability to reproduce crystal structures of small-molecule complexes. Of 161 selected crystal structures of small-molecule guest-host complexes, EUDOC reproduced all these crystal structures with guest structure mass-weighted root mean square deviations (mwRMSDs) of <1.0 Å relative to the corresponding crystal structures. In addition, the average interaction energy of these 161 guest-host complexes (−50.1 kcal/mol) was found to be nearly half of that of 153 previously tested small-molecule-bound protein complexes (−108.5 kcal/mol), according to the interaction energies calculated by EUDOC. 31 of the 161 complexes could not be reproduced with mwRMSDs of <1.0 Å if neighboring hosts in the crystal structure of a guest-host complex were not included as part of the multimeric host system, whereas two of the 161 complexes could not be reproduced with mwRMSDs of <1.0 Å if water molecules were excluded from the host system. These results demonstrate the significant influence of crystal packing on small molecule complexation and suggest that EUDOC is able to predict small-molecule complexes and that it is useful for the design of new materials, molecular sensors, and multimeric inhibitors of protein-protein interactions.


title: A Subcellular Quantitative Proteomic Analysis of Herpes Simplex Virus Type 1-Infected HEK 293T Cells

text: Herpes simplex virus type 1 (HSV-1) is widespread double-stranded DNA (dsDNA) virus that establishes life-long latency and causes diverse severe symptoms. The mechanisms of HSV-1 infection and HSV-1’s interactions with various host cells have been studied and reviewed extensively. Type I interferons were secreted by host cells upon HSV infection and play a vital role in controlling virus proliferation. A few studies, however, have focused on HSV-1 infection without the presence of interferon (IFN) signaling. In this study, HEK 293T cells with low toll-like receptor (TLR) and stimulator of interferon genes protein (STING) expression were infected with HSV-1 and subjected to a quantitative proteomic analysis. By using a subcellular fractionation strategy and high-performance mass spectrometry, a total of 6607 host proteins were quantified, of which 498 proteins were differentially regulated. A bioinformatics analysis indicated that multiple signaling pathways might be involved in HSV-1 infection. A further functional study indicated the role of Interferon-induced transmembrane protein 3 (IFITM3), Coiled-coil-helix-coiled-coil-helix domain-containing protein 2 (CHCHD2), and Tripartite motif-containing protein 27 (TRIM27) in inhibiting viral DNA replication and proliferation. Our data provide a global view of host responses to HSV-1 infection in HEK 293T cells and identify the proteins involved in the HSV-1 infection process.


title: Communication in the Toronto critical care community: important lessons learned during SARS

text: The SARS outbreak in 2003 pushed Toronto's health care system to its limits. Staffing shortages, transmission of SARS within the ICU, and the influx of critically ill SARS patients were some unique challenges to the delivery of critical care. Communication strategies were a key component in the critical care response to SARS. Regular teleconference calls, web-based training and education, and the rapid coordination of research studies were some of the initiatives developed within the Toronto critical care community during the SARS outbreak. Other critical care communities should consider their communication strategies in advance of similar events.


title: Clinical trials and Haemophilia during the COVID‐19’ pandemic: Madrid’s Experience

text: In times of crisis, continuous adaptation is necessary. Communication between all members of a research team is key to adapting the development of clinical trials to the context of the epidemiological crisis of coronavirus. We are accustomed to performing day‐to‐day tasks to fulfil protocol requirements with precision. To this end, we typically have had protocol‐guided face‐to‐face visits and we have followed a detailed sequenced procedure during each visit.


title: We are here for you and ready to hear from you

text: Peer review at JEM during COVID-19


Control electrocardiográfico del intervalo QT mediante dispositivo portátil en pacientes ingresados por COVID-19. Propuesta de protocolo


Virus emergentes y reemergentes: un nuevo reto para la salud mundial del milenio


title: Implementation of rapid SARS-CoV-2 antigenic testing in a laboratory without access to molecular methods: experiences of a general hospital

text: BACKGROUND: The COVID-19 Ag (Antigen) Respi-Strip assay is a new immunochromatographic diagnostic tool recently available for antigenic diagnosis of SARS-CoV-2. The proposed sensitivity is not higher than 60%, but its high specificity allows both quick decisions for the management of patients and confirmation by molecular diagnosis for only negative tests. However, from the first tests performed, we suspected that the sensitivity observed with routine use was much lower than that announced by the manufacturer. MATERIALS AND METHODS: Over a period of one month, we compared the negative results obtained with the COVID-19 Ag Respi-Strip kit with those obtained from qRT-PCR performed in a laboratory qualified for the molecular diagnosis of SARS-CoV-2. All samples tested were naso-pharyngeal smears from UTM-RT medium. RESULTS: Of 774 patients tested, 714 negative samples were sent for confirmation, and 159 were found to be positive by qRT-PCR. The median positive percentage agreement was 23.9% (95% CI: 14.2%-38.2%). The Cohen’s kappa score was 0.35. CONCLUSION: Using this immunochromatographic assay as a triage test did not significantly reduce the number of samples outsourced for COVID-19 confirmation by qRT-PCR. In addition, even if the turn-around time is short, the assay is completely manual, which is not suitable for large volumes of routine samples. The sensitivity of this rapid test is poor, and improvements are needed to enhance its performance.


title: Adaptation of the National Plan for the Prevention and Fight Against Pandemic Influenza to the 2020 COVID-19 Epidemic in France

text: On Tuesday, March 17, 2020, at noon, France became the third European country to impose a nationwide containment policy in the fight against epidemic coronavirus disease 2019 (COVID-19) viral infection. Announcing that the country was at “war,” President Macron called upon all to play a role in mitigating against further development of contagion. This extreme measure never seen before during peace time was the result of adapting not only the French Pandemic Influenza Plan (PIP) being applied to the national context but also real-time clinical, epidemiological, and scientific information about the evolution of COVID-19 infection in the country. The situation was further complicated by local municipal elections and political agendas by populist opinions. Despite mass communication about the importance of individual behavioral attitudes to counter disease propagation, few heeded government advice. Consequently, the situation rapidly deteriorated with increasing number of cases that started to overwhelm health services. As a result, decisive and immediate action was taken by the State for the national public health interest. This report from the field details the timely events that contributed to this extreme policy decision taken by France. A policy decision that other Western democracies have since applied as the pandemic disseminated across the globe.


title: To Lockdown? When to Peak? Will there be an End? A Macroeconomic Analysis on COVID-19 Epidemic in the United States

text: In this paper, we construct an extended SIR model with agents optimally choosing outdoor activities. We calibrate the model and match it to the data from the United States. The model predicts the epidemic in the United States very well. Without government intervention, our simulation shows that the epidemic peaks on 22 March, 2020 and ends on 29 August, 2022. By the end of the epidemic, more than 21 million people will be infected, and the death toll is close to 3.8 million. We further conduct counterfactual experiments to evaluate the effectiveness of different polices against the virus. We find that no single policy can effectively suppress the epidemic, and the most effective policy is a hybrid policy with lockdown and broadening testing. Lockdown policy alone is ineffective in controlling the epidemic as agents would have optimally stayed at home anyway if the infection risk is high even without a lockdown. Broadening testing solely will accelerate the return to normal life as there are fewer infected people hanging around. However, as people do not internalize the social costs of returning to normal life, the epidemic could get even worse. Increasing medical capacity without any other measures only has temporary effects on reducing the death toll. We also find that random testing is too inefficient unless a majority of population is infected.


Sumoylation of the Nucleocapsid Protein of Severe Acute Respiratory Syndrome Coronavirus by Interaction with UBC9


title: Evaluation of the SD Bioline Norovirus rapid immunochromatography test using fecal specimens from Korean gastroenteritis patients

text: Abstract The analytical and clinical performance of a new rapid immunochromatography test, the SD Bioline Norovirus test, was evaluated for the detection of human norovirus in fecal specimens. The analytical performance studies were performed for detection limit, reproducibility, cross-reactivity, and interference. For comparison, 92 norovirus-positive stool samples and 126 norovirus-negative samples for which the results were confirmed by 2 different real-time PCR kits were used. The rapid immunochromatography test detected the equivalent of 4.48×106 copies/mL of the norovirus genome in stool samples. On performing the repeatability/reproducibility test, samples above this concentration all provided positive results (100%) and 97.8% of the samples slightly below this concentration (2.45×106 copies/mL) provided negative results. No cross-reactivity or interference was detected. Positive percent agreement (sensitivity), negative percent agreement (specificity), and overall percent agreement of the rapid immunochromatography test compared with testing by real-time PCR were 90.2%, 100%, and 95.9%, respectively. In addition, the rapid immunochromatography test was completed within 20min. The SD Bioline Norovirus test was, therefore, easier and more rapid to perform and showed excellent reproducibility, no cross-reactivity, no interference, and high agreement compared with real-time PCR. Thus, this test is useful for rapid screening to identity norovirus infection.


title: Clinical characteristics and outcome of respiratory syncytial virus infection among adults hospitalized with influenza-like illness in France

text: Abstract Objectives The aim of this study was to analyse characteristics and outcome of respiratory syncytial virus (RSV) infection in adults hospitalized with influenza-like illness (ILI). Methods Patients hospitalized with ILI were included in this prospective, multicentre study carried out in six French hospitals during three consecutive influenza seasons (2012–2015). RSV and other respiratory viruses were detected by multiplex PCR in nasopharyngeal swabs. Risk factors for RSV infection were identified by backward stepwise logistic regression analysis. Results A total of 1452 patients hospitalized with ILI were included, of whom 59% (861/1452) were >65 years and 83% (1211/1452) had underlying chronic illnesses. RSV was detected in 4% (59/1452), and influenza virus in 39% (566/1452). Risk factors for RSV infection were cancer (adjusted OR 2.1, 95% CI 1.1–4.1, p 0.04), and immunosuppressive treatment (adjusted OR 2.0, 95% CI 1.1–3.8, p 0.03). Patients with RSV had a median length of stay of 9 days (6–25), and 57% of them (30/53) had complications, including pneumonia (23/53, 44%) and respiratory failure (15/53, 28%). Fifteen per cent (8/53) were admitted to an intensive care unit, and the in-hospital mortality rate was 8% (4/53). Pneumonia was more likely to occur in patients with RSV than in patients with RSV-negative ILI (44% (23/53) versus 26% (362/1393), p 0.006) or with influenza virus infection (44% versus 28% (157/560), p 0.02). Conclusion RSV is an infrequent cause of ILI during periods of influenza virus circulation but can cause severe complications in hospitalized adults. Risk factors for RSV detection in adults hospitalized with ILI include cancer and immunosuppressive treatment. Specific immunization and antiviral therapy might benefit patients at risk.


title: Expression cloning of functional receptor used by SARS coronavirus

text: Abstract We have expressed a series of truncated spike (S) glycoproteins of SARS-CoV and found that the N-terminus 14–502 residuals were sufficient to bind to SARS-CoV susceptible Vero E6 cells. With this soluble S protein fragment as an affinity ligand, we screened HeLa cells transduced with retroviral cDNA library from Vero E6 cells and obtained a HeLa cell clone which could bind with the S protein. This cell clone was susceptible to HIV/SARS pseudovirus infection and the presence of a functional receptor for S protein in this cell clone was confirmed by the cell–cell fusion assay. Further studies showed the susceptibility of this cell was due to the expression of endogenous angiotensin-converting enzyme 2 (ACE2) which was activated by inserted LTR from retroviral vector used for expression cloning. When human ACE2 cDNA was transduced into NIH3T3 cells, the ACE2 expressing NIH3T3 cells could be infected with HIV/SARS pseudovirus. These data clearly demonstrated that ACE2 was the functional receptor for SARS-CoV.


title: Spatiotemporal diffusion of influenza A (H1N1): Starting point and risk factors

text: Influenza constitutes a major challenge to world health authorities due to high transmissibility and the capacity to generate large epidemics. This study aimed to characterize the diffusion process of influenza A (H1N1) by identifying the starting point of the epidemic as well as climatic and sociodemographic factors associated with the occurrence and intensity of transmission of the disease. The study was carried out in the Brazilian state of Paraná, where H1N1 caused the largest impact. The units of spatial and temporal analysis were the municipality of residence of the cases and the epidemiological weeks of the year 2009, respectively. Under the Bayesian paradigm, parametric inference was performed through a two-part spatiotemporal model and the integrated nested Laplace approximation (INLA) algorithm. We identified the most likely starting points through the effective distance measure based on mobility networks. The proposed estimation methodology allowed for rapid and efficient implementation of the spatiotemporal model, and provided evidence of different patterns for chance of occurrence and risk of influenza throughout the epidemiological weeks. The results indicate the capital city of Curitiba as the probable starting point, and showed that the interventions that focus on municipalities with greater migration and density of people, especially those with higher Human Development Indexes (HDIs) and the presence of municipal air and road transport, could play an important role in mitigation of effects of future influenza pandemics on public health. These results provide important information on the process of introduction and spread of influenza, and could contribute to the identification of priority areas for surveillance as well as establishment of strategic measures for disease prevention and control. The proposed model also allows identification of epidemiological weeks with high chance of influenza occurrence, which can be used as a reference criterion for creating an immunization campaign schedule.


title: Household animal and human medicine use and animal husbandry practices in rural Bangladesh: risk factors for emerging zoonotic disease and antibiotic resistance

text: Animal antimicrobial use and husbandry practices increase risk of emerging zoonotic disease and antibiotic resistance. We surveyed 700 households to elicit information on human and animal medicine use and husbandry practices. Households that owned livestock (n=265/459, 57.7%) reported using animal treatments 630 times during the previous 6 months; 57.6% obtained medicines, including antibiotics, from drug-sellers. Government animal health care providers were rarely visited (9.7 %), respondents more often sought animal health care from pharmacies and village doctors (70.6% and 11.9% respectively), citing the latter two as less costly and more successful based on past performance. Animal husbandry practices that could promote the transmission of microbes from animals to humans included: the proximity of chickens to humans (50.1% of households reported that the chickens slept in the bedroom); the shared use of natural bodies of water for human and animal bathing (78.3%); the use of livestock waste as fertilizer (60.9%); and gender roles that dictate that females are the primary care takers of poultry and children (62.8%). In the absence of an effective animal health care system, villagers must depend on local animal health care practitioners for treatment of their animals. Suboptimal use of antimicrobials coupled with unhygienic animal husbandry practices are important risk factors for emerging zoonotic disease and resistant pathogens.


title: Time-optimal control strategies in SIR epidemic models

text: Abstract We investigate the time-optimal control problem in SIR (Susceptible-Infected-Recovered) epidemic models, focusing on different control policies: vaccination, isolation, culling, and reduction of transmission. Applying the Pontryagin’s Minimum Principle (PMP) to the unconstrained control problems (i.e. without costs of control or resource limitations), we prove that, for all the policies investigated, only bang–bang controls with at most one switch are admitted. When a switch occurs, the optimal strategy is to delay the control action some amount of time and then apply the control at the maximum rate for the remainder of the outbreak. This result is in contrast with previous findings on the unconstrained problems of minimizing the total infectious burden over an outbreak, where the optimal strategy is to use the maximal control for the entire epidemic. Then, the critical consequence of our results is that, in a wide range of epidemiological circumstances, it may be impossible to minimize the total infectious burden while minimizing the epidemic duration, and vice versa. Moreover, numerical simulations highlighted additional unexpected results, showing that the optimal control can be delayed also when the control reproduction number is lower than one and that the switching time from no control to maximum control can even occur after the peak of infection has been reached. Our results are especially important for livestock diseases where the minimization of outbreaks duration is a priority due to sanitary restrictions imposed to farms during ongoing epidemics, such as animal movements and export bans.


title: Is Pakistan prepared for the COVID‐19 epidemic? A questionnaire‐based survey

text: COVID‐19 is a pandemic that began in China in December 2019. World health organization (WHO) has expressed fears that Pakistan might emerge as the next epicenter of this pandemic. We hypothesize that at present the Pakistani masses are not prepared to face any threat of a looming epidemic. The main aim of this study was to evaluate the basic knowledge of educational and health care workers (HCWs) regarding COVID‐19, its control, and prevention. Knowledge about origin, symptoms, and spread of viral infection was assessed. In this cross‐sectional survey, a self‐designed questionnaire was distributed among 302 HCWs including physicians (10.9%), nurses (1.32%), lab staff (1.65%), and academic individuals including faculty and students (86.42%) of different organizations. Results were analyzed using the χ (2) test. Obtained results validate our null hypothesis that Pakistani masses are not well aware of the COVID‐19 and strategies for the prevention and control of infection. The study concluded that individuals belonging to the front‐line workers and high literacy groups are not prepared for the alarming situation in the country. Effectual implementation of infection control programs should be practiced, and it depends on awareness, training, and cooperation of individuals.


title: Ethical surgical triage of patients with head and neck cancer during the COVID‐19 pandemic

text: BACKGROUND: Coronavirus has serially overtaken our metropolitan hospitals. At peak, patients with acute respiratory distress syndrome may outnumber mechanical ventilators. In our Miami Hospital System, COVID‐19 cases have multiplied for 4 weeks and elective surgery has been suspended. METHODS: An Otolaryngologic Triage Committee was created to appropriately allocate resources to patients. Hospital ethicists provided support. Our tumor conference screened patients for nonsurgical options. Patients were tested twice for coronavirus before performing urgent contaminated operations. N95 masks and protective equipment were conserved when possible. Patients with low‐grade cancers were advised to delay surgery, and other difficult decisions were made. RESULTS: Hundreds of surgeries were canceled. Sixty‐five cases screened over 3 weeks are tabulated. Physicians and patients expressed discomfort regarding perceived deviations from standards, but risk of COVID‐19 exposure tempered these discussions. CONCLUSIONS: We describe the use of actively managed surgical triage to fairly balance our patient's health with public health concerns.


title: Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor

text: The genus Sapovirus belongs to the family Caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. Some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface proteins, such as CD300lf, CD300ld, and junctional adhesion molecule 1 of tight junctions (TJs), as receptors. However, the roles of TJs and their proteins in sapovirus entry have not been examined. In this study, we found that porcine sapovirus (PSaV) significantly decreased transepithelial electrical resistance and increased paracellular permeability early in infection of LLC-PK cells, suggesting that PSaV dissociates TJs of cells. This led to the interaction between PSaV particles and occludin, which traveled in a complex into late endosomes via Rab5- and Rab7-dependent trafficking. Inhibition of occludin using small interfering RNA (siRNA), a specific antibody, or a dominant-negative mutant significantly blocked the entry of PSaV. Transient expression of occludin in nonpermissive Chinese hamster ovary (CHO) cells conferred susceptibility to PSaV, but only for a limited time. Although claudin-1, another TJ protein, neither directly interacted nor was internalized with PSaV particles, it facilitated PSaV entry and replication in the LLC-PK cells. We conclude that PSaV particles enter LLC-PK cells by binding to occludin as a coreceptor in PSaV-dissociated TJs. PSaV and occludin then form a complex that moves to late endosomes via Rab5- and Rab7-dependent trafficking. In addition, claudin-1 in the TJs opened by PSaV infection facilitates PSaV entry and infection as an entry factor. IMPORTANCE Sapoviruses (SaVs) cause severe acute gastroenteritis in humans and animals. Although they replicate in intestinal epithelial cells, which are tightly sealed by apical-junctional complexes, such as tight junctions (TJs), the mechanisms by which SaVs hijack TJs and their proteins for successful entry and infection remain largely unknown. Here, we demonstrate that porcine SaVs (PSaVs) induce early dissociation of TJs, allowing them to bind to the TJ protein occludin as a functional coreceptor. PSaVs then travel in a complex with occludin into late endosomes through Rab5- and Rab7-dependent trafficking. Claudin-1, another TJ protein, does not directly interact with PSaV but facilitates the entry of PSaV into cells as an entry factor. This work contributes to our understanding of the entry of SaV and other caliciviruses into cells and may aid in the development of efficient and affordable drugs to treat SaV infections.


title: The Internet as a Vehicle to Communicate Health Information During a Public Health Emergency: A Survey Analysis Involving the Anthrax Scare of 2001

text: BACKGROUND: The recent public health risks arising from bioterrorist threats and outbreaks of infectious diseases like SARS (Severe Acute Respiratory Syndrome) highlight the challenges of effectively communicating accurate health information to an alarmed public. OBJECTIVE: To evaluate use of the Internet in accessing information related to the anthrax scare in the United States in late 2001, and to strategize about the most effective use of this technology as a communication vehicle during times of public health crises. METHODS: A paper-based survey to assess how individuals obtained health information relating to bioterrorism and anthrax during late 2001.We surveyed 500 randomly selected patients from two ambulatory primary care clinics affiliated with the Brigham and Women's Hospital in Boston, Massachusetts. RESULTS: The response rate was 42%. While traditional media provided the primary source of information on anthrax and bioterrorism, 21% (95% CI, 15% - 27%) of respondents reported searching the Internet for this information during late 2001. Respondents reported trusting information from physicians the most, and information from health websites slightly more than information from any traditional media source. Over half of those searching the Internet reported changing their behavior as a result of information found online. CONCLUSIONS: Many people already look to the Internet for information during a public health crisis, and information found online can positively influence behavioral responses to such crises. However, the potential of the Internet to convey accurate health information and advice has not yet been realized. In order to enhance the effectiveness of public-health communication, physician practices could use this technology to pro-actively e-mail their patients validated information. Still, unless Internet access becomes more broadly available, its benefits will not accrue to disadvantaged populations.


title: T Lymphocytes Promote the Antiviral and Inflammatory Responses of Airway Epithelial Cells

text: HYPOTHESIS: T cells modulate the antiviral and inflammatory responses of airway epithelial cells to human rhinoviruses (HRV). METHODS: Differentiated primary human nasal epithelial cells (HNEC) grown on collagen-coated filters were exposed apically to HRV14 for 6 h, washed thoroughly and co-cultured with anti-CD3/CD28 activated T cells added in the basolateral compartment for 40 h. RESULTS: HRV14 did not induce IFNγ, NOS2, CXCL8 and IL-6 in HNEC, but enhanced expression of the T cell attractant CXCL10. On the other hand, HNEC co-cultured with activated T cells produced CXCL10 at a level several orders of magnitude higher than that induced by HRV14. Albeit to a much lower degree, activated T cells also induced CXCL8, IL-6 and NOS2. Anti-IFNγ antibodies and TNF soluble receptor completely blocked CXCL10 upregulation. Furthermore, a significant correlation was observed between epithelial CXCL10 mRNA expression and the amounts of IFNγ and TNF secreted by T cells. Likewise, increasing numbers of T cells to a constant number of HNEC in co-cultures resulted in increasing epithelial CXCL10 production, attaining a plateau at high IFNγ and TNF levels. Hence, HNEC activation by T cells is induced mainly by IFNγ and/or TNF. Activated T cells also markedly inhibited viral replication in HNEC, partially through activation of the nitric oxide pathway. CONCLUSION: Cross-talk between T cells and HNEC results in activation of the latter and increases their contribution to airway inflammation and virus clearance.


Thrombosis and Coagulopathy in COVID-19 Patients Requiring Extracorporeal Membrane Oxygenation


title: Replication of porcine enteric adenovirusesin vivo

text: The replication of an enteropathogenic porcine adenovirus in the intestinal epithelial cells of naturally and experimentally piglets has been studied by transmission electron microscopy (TEM) and by immunoperoxidase (Ip) staining of paraffin sections. Three types of intranuclear inclusion bodies were observed. Viral particles appeared to be assembled from electron dense crescents which seemed to originate from type II intranuclear inclusion bodies. Virus accumulated in the nucleus of infected cells. It formed paracrystalline arrays in the electron dense centre of the nucleus but was dispersed in the peripheral electron translucent zone. Virus was released from the cells after rupture of the nuclear and plasma membranes. The results are compared with published data on the replication of adenoviruses in cell cultures.


title: Bayesian Reconstruction of Disease Outbreaks by Combining Epidemiologic and Genomic Data

text: Recent years have seen progress in the development of statistically rigorous frameworks to infer outbreak transmission trees (“who infected whom”) from epidemiological and genetic data. Making use of pathogen genome sequences in such analyses remains a challenge, however, with a variety of heuristic approaches having been explored to date. We introduce a statistical method exploiting both pathogen sequences and collection dates to unravel the dynamics of densely sampled outbreaks. Our approach identifies likely transmission events and infers dates of infections, unobserved cases and separate introductions of the disease. It also proves useful for inferring numbers of secondary infections and identifying heterogeneous infectivity and super-spreaders. After testing our approach using simulations, we illustrate the method with the analysis of the beginning of the 2003 Singaporean outbreak of Severe Acute Respiratory Syndrome (SARS), providing new insights into the early stage of this epidemic. Our approach is the first tool for disease outbreak reconstruction from genetic data widely available as free software, the R package outbreaker. It is applicable to various densely sampled epidemics, and improves previous approaches by detecting unobserved and imported cases, as well as allowing multiple introductions of the pathogen. Because of its generality, we believe this method will become a tool of choice for the analysis of densely sampled disease outbreaks, and will form a rigorous framework for subsequent methodological developments.


title: Genomic Analysis of Influenza A Viruses, including Avian Flu (H5N1) Strains

text: This study was designed to conduct genomic analysis in two steps, such as the overall relative synonymous codon usage (RSCU) analysis of the five virus species in the orthomyxoviridae family, and more intensive pattern analysis of the four subtypes of influenza A virus (H1N1, H2N2, H3N2, and H5N1) which were isolated from human population. All the subtypes were categorized by their isolated regions, including Asia, Europe, and Africa, and most of the synonymous codon usage patterns were analyzed by correspondence analysis (CA). As a result, influenza A virus showed the lowest synonymous codon usage bias among the virus species of the orthomyxoviridae family, and influenza B and influenza C virus were followed, while suggesting that influenza A virus might have an advantage in transmitting across the species barrier due to their low codon usage bias. The ENC values of the host-specific HA and NA genes represented their different HA and NA types very well, and this reveals that each influenza A virus subtype uses different codon usage patterns as well as the amino acid compositions. In NP, PA and PB2 genes, most of the virus subtypes showed similar RSCU patterns except for H5N1 and H3N2 (A/HK/1774/1999) subtypes which were suspected to be transmitted across the species barrier, from avian and porcine species to human beings, respectively. This distinguishable synonymous codon usage patterns in non-human origin viruses might be useful in determining the origin of influenza A viruses in genomic levels as well as the serological tests. In this study, all the process, including extracting sequences from GenBank flat file and calculating codon usage values, was conducted by Java codes, and these bioinformatics-related methods may be useful in predicting the evolutionary patterns of pandemic viruses.


title: Innate Immunity to Respiratory Infection in Early Life

text: Early life is a period of particular susceptibility to respiratory infections and symptoms are frequently more severe in infants than in adults. The neonatal immune system is generally held to be deficient in most compartments; responses to innate stimuli are weak, antigen-presenting cells have poor immunostimulatory activity and adaptive lymphocyte responses are limited, leading to poor immune memory and ineffective vaccine responses. For mucosal surfaces such as the lung, which is continuously exposed to airborne antigen and to potential pathogenic invasion, the ability to discriminate between harmless and potentially dangerous antigens is essential, to prevent inflammation that could lead to loss of gaseous exchange and damage to the developing lung tissue. We have only recently begun to define the differences in respiratory immunity in early life and its environmental and developmental influences. The innate immune system may be of relatively greater importance than the adaptive immune system in the neonatal and infant period than later in life, as it does not require specific antigenic experience. A better understanding of what constitutes protective innate immunity in the respiratory tract in this age group and the factors that influence its development should allow us to predict why certain infants are vulnerable to severe respiratory infections, design treatments to accelerate the development of protective immunity, and design age specific adjuvants to better boost immunity to infection in the lung.


title: Estimating Incidence Curves of Several Infections Using Symptom Surveillance Data

text: We introduce a method for estimating incidence curves of several co-circulating infectious pathogens, where each infection has its own probabilities of particular symptom profiles. Our deconvolution method utilizes weekly surveillance data on symptoms from a defined population as well as additional data on symptoms from a sample of virologically confirmed infectious episodes. We illustrate this method by numerical simulations and by using data from a survey conducted on the University of Michigan campus. Last, we describe the data needs to make such estimates accurate.


title: A cross-sectional study of pandemic influenza health literacy and the effect of a public health campaign

text: BACKGROUND: To ascertain the understanding of 2009 pandemic (H1N1) influenza and relevant infection control measures in an emergency department population and to assess the effectiveness of education campaigns in informing the public about the pandemic. METHODS: Questionnaires were administered to patients, visitors, non-clinical staff and volunteers. Data were collected on knowledge, preventative measures, information sources, attitudes to government and media reporting, perceived seriousness, behaviour change and intended compliance with future measures. Results were used to construct an overall knowledge score. RESULTS: There were 252 participants. Traditional forms of mass media (138 [55%]) remained the principal information source. Approximately 70% (176) accurately described mode of transmission and recommended precautions and 68% (175) reported behaviour change because of the pandemic. Gaps in knowledge included failure to identify certain high risk groups. Recall of government campaigns was significantly associated with a higher knowledge score. 60% (151) thought that authorities and media had exaggerated the threat; only 40% (101) would comply with recommended measures in a future pandemic. CONCLUSIONS: The knowledge regarding pandemic influenza was high in this population and positively affected by official campaigns. Pandemic planning should address knowledge gaps and the impression that authorities had exaggerated the public-health threat.


title: Performance of the Alere i RSV assay for point-of-care detection of respiratory syncytial virus in children

text: BACKGROUND: Respiratory syncytial virus (RSV) is the most important cause of severe acute respiratory tract infection in young children. Alere i RSV is a novel molecular rapid test which identifies respiratory syncytial virus in less than 13 min. METHODS: We evaluated the clinical performance of the Alere i RSV assay in a pediatric point-of-care setting during winter season 2016 / 2017. Test results from 518 nasopharyngeal swab samples were compared to a real-time reverse transcription PCR reference standard. RESULTS: The overall sensitivity and specificity of the Alere i RSV test assay was 93% (CI(95) 89% – 96%) and 96% (CI(95) 93% – 98%), respectively. Alere i RSV performed well in children of all age groups. An optimal sensitivity of 98% (CI(95) 94% - 100%) and specificity of 96% (CI(95) 90% - 99%) was obtained in children < 6 months. In children ≥ 2 years, sensitivity and specificity remained at 87% (CI(95) 73% – 96%) and 98% (CI(95) 92% – 100%), respectively. False negative Alere i RSV test results mostly occurred in samples with low viral load (mean CT value 31.1; CI(95) 29.6 – 32.6). The Alere i RSV assay is easy to use and can be operated after minimal initial training. Test results are available within 13 min, with most RSV positive samples being identified after approximately 5 min. CONCLUSION: The Alere i RSV assay has the potential to facilitate the detection of RSV in pediatric point-of-care settings. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12879-017-2855-1) contains supplementary material, which is available to authorized users.


title: Association of functional polymorphisms in the MxA gene with susceptibility to enterovirus 71 infection

text: Myxovirus resistance A (MxA) is an antiviral protein induced by type I interferons α and β (IFN-α and IFN-β) that can inhibit virus replication. We examined whether the MxA polymorphisms were related to the risk and severity of enterovirus 71 (EV71) infection in Chinese populations. The MxA C-123A and G-88T polymorphisms were genotyped in two independent case–control populations in China by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) analysis. Multivariate logistic regression analysis was used to calculate adjusted odds ratios (ORs) and 95 % confidence intervals (95 % CIs). MxA messenger RNA was quantified by real-time quantitative PCR in peripheral blood mononuclear cells (PBMCs) from 45 healthy children and 19 patients with EV71 infection. Significantly decreased susceptibility to EV71 infection was observed for the -123A allele and -88T allele carriers, with ORs (95 % CIs) estimated as 0.56 (0.39–0.81) and 0.64 (0.47–0.88), respectively, in the northern population. This association was confirmed in the southern population, with ORs (95 % CIs) estimated as 0.58 (0.38–0.89) and 0.67(0.47–0.95), respectively. The A(-) (123)T(-) (88) haplotype was also significantly associated with lower risk of EV71 infection in both the northern (OR = 0.62; 95 % CI = 0.44–0.85) and the southern population (OR = 0.63; 95 % CI = 0.43–0.92). Furthermore, we observed higher MxA messenger RNA levels in IFNβ1a-stimulated PBMCs from the -123A or -88T allele carriers compared with that from nocarriers. Our findings suggest that polymorphisms in the MxA promoter may play a role in mediating the susceptibility to EV71 infection in Chinese population.


title: Evolutionary selection against short nucleotide sequences in viruses and their related hosts

text: Viruses are under constant evolutionary pressure to effectively interact with the host intracellular factors, while evading its immune system. Understanding how viruses co-evolve with their hosts is a fundamental topic in molecular evolution and may also aid in developing novel viral based applications such as vaccines, oncologic therapies, and anti-bacterial treatments. Here, based on a novel statistical framework and a large-scale genomic analysis of 2,625 viruses from all classes infecting 439 host organisms from all kingdoms of life, we identify short nucleotide sequences that are under-represented in the coding regions of viruses and their hosts. These sequences cannot be explained by the coding regions’ amino acid content, codon, and dinucleotide frequencies. We specifically show that short homooligonucleotide and palindromic sequences tend to be under-represented in many viruses probably due to their effect on gene expression regulation and the interaction with the host immune system. In addition, we show that more sequences tend to be under-represented in dsDNA viruses than in other viral groups. Finally, we demonstrate, based on in vitro and in vivo experiments, how under-represented sequences can be used to attenuated Zika virus strains.


title: Foodborne viruses

text: Among the wide variety of viral agents liable to be found as food contaminants, noroviruses and hepatitis A virus are responsible for most well characterized foodborne virus outbreaks. Additionally, hepatitis E virus has emerged as a potential zoonotic threat. Molecular methods, including an ISO standard, are available for norovirus and hepatitis A virus detection in foodstuffs, although the significance of genome copy detection with regard to the associated health risk is yet to be determined through viability assays. More precise and rapid methods for early foodborne outbreak investigation are being developed and they will need to be validated versus the ISO standard. In addition, protocols for next-generation sequencing characterization of outbreak-related samples must be developed, harmonized and validated as well.


title: Genotypes and subtypes of Cryptosporidium spp. in neonatal calves in Northern Ireland

text: Cryptosporidium spp. in diarrheic calves less than 30 days old from farms across Northern Ireland were examined over a year period by microscopic, genotyping, and subtyping techniques to characterize the transmission dynamics. Cryptosporidium oocysts were detected in 291 of 779 (37.4%) animals. The prevalence rates of rotavirus, coronavirus, and Escherichia coli K99+ were lower as seen in 242 of 806 (30.0%), 46/806 (5.7%), and 16/421 (3.8%) of animals, respectively. Of the 224 Cryptosporidium-positive specimens available for molecular analysis, Cryptosporidium parvum was identified in 213 (95.1%) specimens, Cryptosporidium bovis in eight (3.6%), and Cryptosporidium deer-like genotype in three (1.3%). Sequence analysis of the 60-kDa glycoprotein gene identified 16 IIa subtypes and a new subtype family, with 120 of the 216 (55.6%) positive specimens having the subtype IIaA18G3R1. Eight of the IIa subtypes were previously seen in humans in Northern Ireland. Several subtypes were temporally or geographically unique. The genetic diversity in calves in Northern Ireland was much greater than that reported from other areas. This work demonstrates the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. in calves and humans.


title: COVID-19 Pandemic and the Implications for Orthopaedic and Neurosurgery Residents and Fellows on Spine Rotations

text: Abstract Background The COVID-19 global pandemic has caused unprecedented levels of strain on the United States healthcare and its workforce. Orthopaedic and neurosurgery residents and fellows, as part of this workforce have also experienced some of the uncertainty and stress caused by this pandemic. Concerns exist regarding the effects of the pandemic on spine surgery education due to the cancellation of all elective surgeries. Current Context We explore how this pandemic is affecting orthopaedic and neurosurgery residents and fellows and their spine surgery education and experience. We also examined measures taken by the residency and fellowship programs to protect their residents and fellows, and measures taken by regulatory agencies like the ACGME and the ABOS to give programs some flexibility during these difficult times. Conclusion Orthopaedic and neurosurgery residents and fellows are often on the front lines of patient care. Programs have to ensure adequate resources and training, supervision, and work hour requirements are met. Residents and fellows need to be ready to assist with management of COVID-19 patients if necessary. Residency programs and spine surgery fellowships need to use objective metrics to assess the impact of the pandemic on the spine surgery education of their residents and fellows in order to address any potential area of weakness caused by the decreased exposure to spine surgery.


title: Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid

text: The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies (mAbs) targeting N protein of PEDV and analyzed their applications on enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), western blot assay, and flow cytometry assay. The results demonstrated that all these six mAbs were IgG1 isotype and κ chain. Among these six mAbs, 3F12 recognizes a linear epitope (VAAVKDALKSLGI) while the other five mAbs recognize different conformational epitopes formed by a specific peptide fragment or the full length of N protein. The functional analysis showed that all these six mAbs were applicable to ELISA, western blot, IFA, and flow cytometry assay. In conclusion, we developed six mAbs against PEDV-N protein to facilitate the early detection of PEDV infection using ELISA, western blot, IFA, and flow cytometry. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09702-5) contains supplementary material, which is available to authorized users.


Single-use Bronchoscopes: Applications in COVID-19 Pandemic