title: Fourteen-day inhalation study in rats, using aged and diluted sidestream smoke from a reference cigarette I. Inhalation toxicology and histopathology

text: Abstract Sprague-Dawley rats were exposed 6 hr per day for 14 consecutive days to aged and diluted sidestream smoke (ADSS), used as a surrogate for Environmental Tobacco Smoke (ETS), at concentrations of 0.1 (typical), 1 (extreme), or 10 (exaggerated) mg of particulates per cubic meter. Animals were exposed nose-only, inside whole-body chambers, to ADSS from the 1R4F reference cigarette. End-points included histopathology, CO-oximetry, plasma nicotine and cotinine, clinical pathology, and organ and body weights. The only pathological response observed was slight to mild epithelial hyperplasia and inflammation in the most rostral part of the nasal cavity, in the high-exposure group only. No effects were noted at medium or low exposures. The minimal changes noted were reversible, using a subgroup of animals kept without further treatment for an additional 14 days. Overall, the end-points used in the study demonstrated that there was no detectable biological activity of ADSS at typical or even 10-fold ETS concentrations and that the activity was only minimal at very exaggerated concentrations (particle concentrations 100 times higher than typical real-world concentrations).


title: Clinical features and chest CT findings of three cases of 2019 novel coronavirus (COVID-2019) pneumonia

text: BACKGROUND: Since December 2019, a new type of acute respiratory disease caused by the 2019 novel coronavirus (COVID-2019) appeared to Wuhan, China. Currently, positive of virus nucleic acid in respiratory or blood samples is the gold standard for the diagnosis of the disease, but the nucleic acid test may be false negative. Chest CT examination plays an important role in diagnosis of COVID-2019 pneumonia. CASE PRESENTATION: The clinical manifestations, laboratory examination, and chest CT characteristics of 3 patients with COVID-2019 pneumonia were reported. The mainly clinical manifestation of the 3 patients was fever. white blood cell, neutrophil cell and lymphocyte cell count can be normal, only C-Reactive Protein slightly elevated. Real-time fluorescence polymerase chain reaction of throat swab detection can be negative. Chest CT showed multifocal ground-glass opacities (GGO) in the peripheral of bilateral lungs. GGOs with consolidation, air bronchogram, vascular enlargement and halo sign were be found. CONCLUSIONS: In the early stages of COVID-2019 pneumonia, the laboratory parameters may be normal, the nucleic acid test may be false negative. Chest CT examination may be helpful for diagnosis of COVID-2019 pneumonia.


title: Interaction of cationic carbosilane dendrimers and their complexes with siRNA with erythrocytes and red blood cell ghosts

text: Abstract We have investigated the interactions between cationic NN16 and BDBR0011 carbosilane dendrimers with red blood cells or their cell membranes. The carbosilane dendrimers used possess 16 cationic functional groups. Both the dendrimers are made of water-stable carbon–silicon bonds, but NN16 possesses some oxygen–silicon bonds that are unstable in water. The nucleic acid used in the experiments was targeted against GAG-1 gene from the human immunodeficiency virus, HIV-1. By binding to the outer leaflet of the membrane, carbosilane dendrimers decreased the fluidity of the hydrophilic part of the membrane but increased the fluidity of the hydrophobic interior. They induced hemolysis, but did not change the morphology of the cells. Increasing concentrations of dendrimers induced erythrocyte aggregation. Binding of short interfering ribonucleic acid (siRNA) to a dendrimer molecule decreased the availability of cationic groups and diminished their cytotoxicity. siRNA–dendrimer complexes changed neither the fluidity of biological membranes nor caused cell hemolysis. Addition of dendriplexes to red blood cell suspension induced echinocyte formation.


title: Angiotensin-converting enzyme 2 prevents lipopolysaccharide-induced rat acute lung injury via suppressing the ERK1/2 and NF-κB signaling pathways

text: Acute respiratory distress syndrome (ARDS) caused by severe sepsis remains a major challenge in intensive care medicine. ACE2 has been shown to protect against lung injury. However, the mechanisms of its protective effects on ARDS are largely unknown. Here, we report that ACE2 prevents LPS-induced ARDS by inhibiting MAPKs and NF-κB signaling pathway. Lentiviral packaged Ace2 cDNA or Ace2 shRNA was intratracheally administrated into the lungs of male SD rats. Two weeks after gene transfer, animals received LPS (7.5 mg/Kg) injection alone or in combination with Mas receptor antagonist A779 (10 μg/Kg) or ACE2 inhibitor MLN-4760 (1 mg/Kg) pretreatment. LPS-induced lung injury and inflammatory response were significantly prevented by ACE2 overexpression and deteriorated by Ace2 shRNA. A779 or MLN-4760 pretreatment abolished the protective effects of ACE2. Moreover, overexpression of ACE2 significantly reduced the Ang II/Ang-(1-7) ratio in BALF and up-regulated Mas mRNA expression in lung, which was reversed by A779. Importantly, the blockade of ACE2 on LPS-induced phosphorylation of ERK1/2, p38 and p50/p65 was also abolished by A779. Whereas, only the ERK1/2 inhibitor significantly attenuated lung injury in ACE2 overexpressing rats pretreated with A779. Our observation suggests that AEC2 attenuates LPS-induced ARDS via the Ang-(1-7)/Mas pathway by inhibiting ERK/NF-κB activation.


title: Do we have enough evidence how seasonal influenza is transmitted and can be prevented in hospitals to implement a comprehensive policy?

text: PURPOSE: To identify if there is enough evidence at low risk-of-bias to prevent influenza transmission by vaccinating health-care workers (HCWs), patients and visitors; screening for laboratory-proven influenza all entering hospitals; screening asymptomatic individuals; identifying influenza supershedders; hand-washing and mask-wearing by HCWs, patients and visitors; and cleaning hospital rooms and equipment. PRINCIPAL RESULTS: Vaccination reduces influenza episodes of vaccinated (4.81/100 HCW) compared to unvaccinated (7.54/100) HCWs/influenza season. A Cochrane review found for inactivated vaccines the Number Needed to Vaccinate (NNV) = 71 (95%CI 64%, 80%) for adults 18–60 (same age as HCWs) to prevent laboratory-proven influenza. There are no RCTs of screening HCWs, patients, visitors and influenza supershedders to prevent transmission. None of four RCTs of HCWs mask-wearing (two directly observed, two not) showed an effect because they were underpowered either due to small size or low circulation of influenza. Hospital rooms and equipment can effectively be cleaned of influenza by many chemicals and hydrogen peroxide vapor machines but the cleaning cycle needs shortening to increase the likelihood of adoption. MAJOR CONCLUSIONS: HCW vaccination is a partial solution with current vaccination levels. There are no RCTs of screening HCWs, patients and visitors demonstrating preventing influenza transmission. Only one study costed furloughing HCWs with influenza and no RCTs have identified benefits of isolating influenza supershedders. RCTs of directly- and electronically continuously-observed mask-wearing and hand-hygiene and RCTs of incentives for meticulous hygiene are required. RCTs of engineering solutions (external venting, frequent room air changes) are needed. A wide range of chemicals effectively cleans hospital rooms and equipment from influenza. Hydrogen peroxide vapor is effective against influenza and a wide range of bacterial pathogens with patient room changes, and clean areas cleaners do not clean but its cleaning cycle needs shortening to increase the likelihood of adoption of cleaning rooms vacated by influenza patients.


title: Dynamical response of multi-patch, flux-based models to the input of infected people: Epidemic response to initiated events

text: Abstract The time course of an epidemic can be modeled using the differential equations that describe the spread of disease and by dividing people into “patches” of different sizes with the migration of people between these patches. We used these multi-patch, flux-based models to determine how the time course of infected and susceptible populations depends on the disease parameters, the geometry of the migrations between the patches, and the addition of infected people into a patch. We found that there are significantly longer lived transients and additional “ancillary” epidemics when the reproductive rate R is closer to 1, as would be typical of SARS (Severe Acute Respiratory Syndrome) and bird flu, than when R is closer to 10, as would be typical of measles. In addition we show, both analytical and numerical, how the time delay between the injection of infected people into a patch and the corresponding initial epidemic that it produces depends on R.


Impacto de la pandemia COVID-19 en los tratamientos inmunomoduladores/inmunosupresores en dermatología: actitudes de los pacientes y los dermatólogos en Argentina


title: A deep dive into testing and management of COVID-19 for Australian high performance and professional sport

text: Abstract The purpose of testing for any communicable disease is to support clinicians in the diagnosis and management of individual patients and to describe transmission dynamics. The novel coronavirus is formally named SARS-CoV-2 and the clinical disease state resulting from an infection is known as COVID-19. Control of the COVID-19 pandemic requires clinicians, epidemiologists, and public health officials to utilise the most comprehensive, accurate and timely information available to manage the rapidly evolving COVID-19 environment. High performance sport is a unique context that may look towards comprehensive testing as a means of risk mitigation. Characteristics of the common testing options are discussed including the circumstances where additional testing may be of benefit and considerations for the associated risks. Finally, a review of the available technology that could be considered for use by medical staff at the point of care (PoC) in a high-performance sporting context is included.


title: No Evidence of Viral Transmission following Long-Term Implantation of Agarose Encapsulated Porcine Islets in Diabetic Dogs

text: We have previously described the use of a double coated agarose-agarose porcine islet macrobead for the treatment of type I diabetes mellitus. In the current study, the long-term viral safety of macrobead implantation into pancreatectomized diabetic dogs treated with pravastatin (n = 3) was assessed while 2 dogs served as nonimplanted controls. A more gradual return to preimplant insulin requirements occurred after a 2nd implant procedure (days 148, 189, and >652) when compared to a first macrobead implantation (days 9, 21, and 21) in all macrobead implanted animals. In all three implanted dogs, porcine C-peptide was detected in the blood for at least 10 days following the first implant and for at least 26 days following the second implant. C-peptide was also present in the peritoneal fluid of all three implanted dogs at 6 months after 2nd implant and in 2 of 3 dogs at necropsy. Prescreening results of islet macrobeads and culture media prior to transplantation were negative for 13 viruses. No evidence of PERV or other viral transmission was found throughout the study. This study demonstrates that the long-term (2.4 years) implantation of agarose-agarose encapsulated porcine islets is a safe procedure in a large animal model of type I diabetes mellitus.


title: The Genome Organization of the Nidovirales: Similarities and Differences between Arteri-, Toro-, and Coronaviruses

text: Abstract Viruses in the families Arteriviridae and Coronaviridae have enveloped virions which contain nonsegmented, positive-stranded RNA, but the constituent genera differ markedly in genetic complexity and virion structure. Nevertheless, there are striking resemblances among the viruses in the organization and expression of their genomes, and sequence conservation among the polymerase polyproteins strongly suggests that they have a common ancestry. On this basis, the International Committee on Taxonomy of Viruses recently established a new order, Nidovirales, to contain the two families. Here, the common traits and distinguishing features of the Nidovirales are reviewed.


COVID‐19 era: A chance to learn something new about monitoring psoriatic patients in biological therapy


title: Effect of Trade Openness on Food Security in the EU: A Dynamic Panel Analysis

text: The problem of food insecurity is growing across the world, including economically developed countries. In Europe, the question is not just about the total supply of foods, but it includes even the accessibility of prices and their nutritional and qualitative adequacy. In this context many countries recognize the importance of trade policies to ensure adequate levels of food security. The aim of this work was to analyze the impact of trade openness on the level of food security in European countries, using a dynamic panel analysis with the generalized method of moments (GMM) approach. We selected two different indicators of food security (average protein supply, average dietary energy supply adequacy) capable of offering information both on the quantity and on the nutritional quality of the food supply. In order to improve the robustness of the empirical results, we developed three different regressions, with three trade openness indicators (trade openness, tariff, globalization) for each food security indicator. The results showed that commercial opening has, on average, a statistically significant net positive impact on the food security of European countries. Additional results indicate that also economic development, together with the importance of the agricultural sector, can improve food security levels.


A newly designed real-time RT-PCR for SAFV detects SAFV-2 and SAFV-3 in the respiratory tracts of ill children during 2011


title: Effectiveness of Loaded Sit-to-Stand Resistance Exercise for Children With Mild Spastic Diplegia: A Randomized Clinical Trial

text: Abstract Liao H-F, Liu Y-C, Liu W-Y, Lin Y-T. Effectiveness of loaded sit-to-stand resistance exercise for children with mild spastic diplegia: a randomized clinical trial. Objective To investigate effectiveness of a functional strengthening program, the loaded sit-to-stand (STS) resistance exercise, for children with cerebral palsy (CP). Design A single-blind, randomized block design. Setting STS exercises were carried out at the children’s homes. Participants Twenty children (12 boys, 8 girls; age range, 5−12y) with spastic diplegia CP and classified by the Gross Motor Function Classification System as level I or II were stratified by their severity and age and randomly allocated into either the experimental or control group. Intervention Both groups received their regular physical therapy. The experimental group underwent loaded STS exercise 3 times a week for 6 weeks. Main Outcome Measures Goal dimension scores of the Gross Motor Function Measure (GMFM), gait speed, 1 repetition maximum (1-RM) of the loaded STS, isometric strength of knee extensor, and Physiological Cost Index (PCI). The outcome measures were conducted at the beginning and end of the 6-week study. Results After loaded STS exercise, the experimental group showed statistically significant differences in GMFM goal dimension scores, 1-RM STS, and PCI from the control group. The changes in gait speed and isometric strength of the knee extensor did not differ significantly between the 2 groups. Conclusions After the loaded STS exercise, children with mild spastic diplegia improved their basic motor abilities, functional muscle strength, and walking efficiency.


Central Apnea in a COVID19 Pediatric Patient


title: Clinical course of severe and critical COVID-19 in hospitalized pregnancies: a US cohort study

text: Abstract Background The COVID-19 pandemic has had an impact on healthcare systems around the world with 3.0 million infected and 208,000 resultant mortalities as of this writing. Information regarding infection in pregnancy is still limited. Objectives To describe the clinical course of severe and critical infection in hospitalized pregnant women with positive laboratory testing for SARS-CoV2. Study Design This is a cohort study of pregnant women with severe or critical COVID-19 infection hospitalized at 12 US institutions between March 5, 2020 and April 20, 2020. Severe infection was defined according to published criteria by patient reported dyspnea, respiratory rate > 30 per minute, blood oxygen saturation ≤ 93% on room air, partial pressure of arterial oxygen to fraction of inspired oxygen <300 and/or lung infiltrates >50% within 24 to 48 hours on chest imaging. Critical disease was defined by respiratory failure, septic shock, and/or multiple organ dysfunction or failure. Women were excluded if they had presumed COVID-19 infection but laboratory testing was negative. The primary outcome was median duration from hospital admission to discharge. Secondary outcomes included need for supplemental oxygen, intubation, cardiomyopathy, cardiac arrest, death, and timing of delivery. The clinical courses are described by the median disease day on which these outcomes occurred after the onset of symptoms. Treatment and neonatal outcomes are also reported. Results Of 64 pregnant women hospitalized with COVID-19, 44 (69%) had severe and 20 (31%) critical disease. The following pre-existing comorbidities were observed: 25% had a pulmonary condition, 17% had cardiac disease and the mean BMI was 34 kg/m2. Gestational age at symptom onset was at a mean 29 ±6 weeks and at hospital admission a mean of 30 ±6 weeks, on a median day of disease 7 since first symptoms. Eighty-one percent of women were treated with hydroxychloroquine; 9% of women with severe disease and 65% of women with critical disease received remdesivir. All women with critical disease received either prophylactic or therapeutic anticoagulation during their admission. The median duration of hospital stay was 6 days (6 days for severe, 10.5 days for critical, p=0.01). For those who required it, intubation usually occurred around day 9, and peak respiratory support for women with severe disease occurred on day 8. In women with critical disease, prone positioning was performed in 20% of cases, the rate of ARDS was 70%, and re-intubation was necessary in 20%. There was one case of maternal cardiac arrest, but no cases of cardiomyopathy and no maternal deaths. Thirty-two (50%) women in this cohort delivered during their COVID-19 hospitalization (34% of severe and 85% of critical women). Eighty-eight percent (15/17) of pregnant women with critical COVID-19 who delivered during their disease course were delivered preterm, 94% of them via cesarean; in all, 75% (15/20) of critically ill women delivered preterm. There were no stillbirths or neonatal deaths, or cases of vertical transmission. Conclusion In hospitalized pregnant women with severe or critical COVID-19 infection, admission typically occurred about 7 days after symptom onset, and the duration of hospitalization was 6 days (6 severe versus 12 critical). Critically ill women had a high rate of ARDS, and there was one case of cardiac arrest, but there were no cases of cardiomyopathy, or maternal mortality. Hospitalization for severe or critical COVID-19 infection resulted in delivery during the course of infection in 50% of this cohort, usually in the third trimester. There were no perinatal deaths in this cohort.


title: Recombination

text: Genetic recombination of viruses is a commonly found phenomenon in both DNA and RNA viruses. By exchanging of fragments of genetic material viruses gain important means leading to both variability and also to the repair of their genome. Biochemically, the processes of DNA and RNA recombination are different reflecting the specifics of DNA versus RNA replication as well as their use inside the cell. A variety of recombination mechanisms are discussed and illustrated by examples taken from specific viral species.


title: TIM-family Proteins Promote Infection of Multiple Enveloped Viruses through Virion-associated Phosphatidylserine

text: Human T-cell Immunoglobulin and Mucin-domain containing proteins (TIM1, 3, and 4) specifically bind phosphatidylserine (PS). TIM1 has been proposed to serve as a cellular receptor for hepatitis A virus and Ebola virus and as an entry factor for dengue virus. Here we show that TIM1 promotes infection of retroviruses and virus-like particles (VLPs) pseudotyped with a range of viral entry proteins, in particular those from the filovirus, flavivirus, New World arenavirus and alphavirus families. TIM1 also robustly enhanced the infection of replication-competent viruses from the same families, including dengue, Tacaribe, Sindbis and Ross River viruses. All interactions between TIM1 and pseudoviruses or VLPs were PS-mediated, as demonstrated with liposome blocking and TIM1 mutagenesis experiments. In addition, other PS-binding proteins, such as Axl and TIM4, promoted infection similarly to TIM1. Finally, the blocking of PS receptors on macrophages inhibited the entry of Ebola VLPs, suggesting that PS receptors can contribute to infection in physiologically relevant cells. Notably, infection mediated by the entry proteins of Lassa fever virus, influenza A virus and SARS coronavirus was largely unaffected by TIM1 expression. Taken together our data show that TIM1 and related PS-binding proteins promote infection of diverse families of enveloped viruses, and may therefore be useful targets for broad-spectrum antiviral therapies.


title: Effect of Graded Concentrations of Gossypol on Calf Performance: Toxicological and Pathological Considerations 1

text: Abstract The effects of feeding diets containing 0, 100, 200, 400, or 800ppm of free gossypol were examined in 60 Holstein bull calves from 1 to 120 d of age. The concentrations of free gossypol were varied by varying amounts of cottonseed meals from three different sources. Cottonseed meal totaled 31% of each treatment ration. Feed consumption, BW, and blood parameters were collected on all calves at 30-d intervals throughout the trial. There were no significant differences between the groups for feed consumption, BW, or average daily gain. Changes in the group means for hematology and chemistry variables examined were modest and insufficient to distinguish diagnostically between safe and unsafe free gossypol concentrations for the different groups. Clinical evidence of disease was limited to the calves fed 400 or 800ppm of free gossypol after 90 d of age. One calf in the group fed 400ppm and 4 in the group fed 800ppm died as a result of circulatory failure associated with gossypol consumption. We conclude from this study that a ration containing up to 200ppm of free gossypol is safe, 400ppm of free gossypol is toxic, and 800ppm of free gossypol results in death losses. These results are compatible with previously reported naturally occurring toxicological outbreaks.


title: Immunology of the Porcine Respiratory Disease Complex

text: Respiratory disease is a serious economic problem for swine producers and veterinarians worldwide. The term porcine respiratory disease complex (PRDC) has been used to describe the respiratory disease pattern that has recently emerged. PRDC is multifactorial, involving management factors and multiple pathogens. This article summarizes the pathogenesis and immune response to some of the common respiratory pathogens associated with PRDC, including interactions that lead to the severe respiratory disease associated with PRDC.


title: Experience with Cidofovir as an adjunctive therapy in a patient of adenovirus-induced macrophage activation syndrome in systemic arthritis

text: A 5-year-old female child, with known systemic juvenile idiopathic arthritis diagnosed at 18 months of age (on low dose Prednisolone + Methotrexate + Leflunomide + Tocilizumab), presented with fever for 1 day, vomiting, drowsiness followed by seizures. On admission to PICU, she was drowsy, tachycardic, tachypneic, with rashes, and hepatosplenomegaly. Lab findings showed thrombocytopenia, leucopenia, low ESR, normal CRP, elevated liver enzymes, high ferritin, LDH, and triglycerides suggestive of macrophage activation syndrome (MAS). Chest X-ray showed left basal pneumonia and DNA PCR of throat swab revealed adenovirus. She was diagnosed as adenovirus-triggered MAS and was initiated on pulse methylprednisolone (6 mg/kg). Because of suboptimal response after 2 doses, manifested by increasing drowsiness, further fall in platelets and rising ferritin, methylprednisolone dosage was increased to 30 mg/kg/day with the addition of oral cyclosporine (4 mg/kg/day). In view of worsening of the chest X-ray and increasing oxygen requirement, Cidofovir infusion (1 mg/kg thrice weekly) was also started simultaneously considering increased activity of the adenoviral infection concurrent to immunosuppression. Within 48 h, the child showed signs of recovery with improved consciousness, lower oxygen requirements, and improving lab parameters. She was discharged after 3 weeks of IV Cidofovir, on oral prednisolone and cyclosporine. To the best of our knowledge, this is the first reported use of Cidofovir in adenovirus-induced MAS. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10067-020-05133-0) contains supplementary material, which is available to authorized users.


title: Prevalence of Leishmania infantum and co-infections in stray cats in northern Italy

text: Abstract Stray cats in the city of Milan, Italy, were tested for Leishmania infantum and other selected infections. Twenty-seven cats (30.0%) were seroreactive by indirect fluorescent antibody test (IFAT), with an antibody titer of 1:40 for 16 (17.7%) cats and 1:80 (cut-off for feline L. infantum infection) for 11 (12.2%) cats. One blood (1.1%) and one popliteal lymph node (1.1%) sample tested positive by real-time polymerase chain reaction; no oculoconjunctival swabs tested positive. Feline immunodeficiency virus, feline leukemia virus, and feline coronavirus (FCoV) seroprevalence determined by enzyme-linked immunosorbent assay was 6.1, 6.1, and 39.0%, respectively. Toxoplasma gondii, Bartonella henselae, and Chlamydophila felis prevalence determined by IFAT was 29.3, 17.1, and 17.1%, respectively. The frequency of seroreactivity to L. infantum was significantly higher in FCoV-seropositive cats (OR=4.4, P =0.04). L. infantum-infected stray cats in Milan have a high seropositivity rate, comparable to that of cats in areas endemic for leishmaniosis.


Limiting spread of COVID-19 from cruise ships - lessons to be learnt from Japan


A Pinch of Prevention is Worth a Pound of Cure: Proactive Dentistry in the Wake of COVID-19.


REACTing: the French response to infectious disease crises


title: Could SARS-CoV-2 be transmitted via speech droplets?

text: Speaking may be a primary mode of transmission of SARS-CoV-2. Considering that reports of asymptomatic transmission account for 50-80% of COVID-19 cases and that saliva has peak viral loads at time of patient presentation, droplet emission while speaking could be a significant factor driving transmission and warrants further study. We used a planar beam of laser light passing through a dust-free enclosure to detect saliva droplets emitted while speaking. We found that saying the words 'Stay Healthy' generates thousands of droplets that are otherwise invisible to the naked eye. A damp homemade cloth face mask dramatically reduced droplet excretion, with none of the spoken words causing a droplet rise above the background. Our preliminary findings have important implications for pandemic mitigation efforts.


title: SARS, MERS and COVID-19 among Healthcare Workers: A Narrative Review

text: Abstract In the recent two decades, three global viral infectious diseases, severe acute respiratory syndrome (SARS), middle east respiratory syndrome (MERS), and coronavirus disease (COVID-19), have occurred worldwide. SARS occurred in November 2002, causing 8,096 infected cases, as well as 774 deaths. MERS occurred in June, 2012, causing 2,519 confirmed cases, along with 866 associated deaths. COVID-19 occurred in December 2019, as of 30 April 2020, a total of 3,024,059 clinical cases have been reported, including 208,112 deaths. Healthcare workers (HCWs) need to be in close contact with these virus-infected patients and their contaminated environments at work, thus leading to be infected in some of them, even a few of them are died in line of duty. In this review, we summarized the infection status of HCWs during the outbreak of SARS, MERS and COVID-19, with in-depth discussion, hoping to provoke sufficient attention to the HCWs infection status by more people.


title: 肿瘤合并新型冠状病毒感染的临床特征与应对策略

text: Since mid-December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has outbroken in Wuhan, Hubei Province, China, and spread rapidly to other provinces in China and dozens of countries and regions around the world, becoming the Public Health Emergency of International Concern (Public Health Emergency of International Concern). SARS-CoV-2 can mainly transmit by droplets or close contact, and is generally susceptible in the crowd. Tumor patients are at high risk of this pathogen because of their impaired immune function. Identifying tumor patients with 2019 novel coronavirus disease (COVID-19) early, and understanding its distribution characteristics can help to improve the cure rate of patients, and better control the epidemic and development of SARS-CoV-2 much better. With comprehensive analysis of relevant literature, this paper reviews the clinical characteristics of neoplastic patients with COVID-19, and puts forward some suggestions on how to deal with this epidemic.


title: The use of synthetic polymers for delivery of therapeutic antisense oligodeoxynucleotides

text: Abstract Developed over the past two decades, the antisense strategy has become a technology of recognised therapeutic potential, and many of the problems raised earlier in its application have been solved to varying extents. However, the adequate delivery of antisense oligodeoxynucleotides to individual cells remains an important and inordinately difficult challenge. Synthetic polymers appeared on this scene in the middle 1980s, and there is a surprisingly large variety used or proposed so far as agents for delivery of oligodeoxynucleotides. After discussing the principles of antisense strategy, certain aspects of the ingestion of macromolecules by cells, and the present situation of delivery procedures, this article analyses in detail the attempts to use synthetic polymers as carrier matrices and/or cell membrane permeabilisation agents for delivery of antisense oligodeoxynucleotides. Structural aspects of various polymers, as well as the results, promises and limitations of their use are critically evaluated.


title: Generation of Recombinant Antibodies Against Toxins and Viruses by Phage Display for Diagnostics and Therapy

text: Antibody phage display is an in vitro technology to generate recombinant antibodies. In particular for pathogens like viruses or toxins, antibody phage display is an alternative to hybridoma technology, since it circumvents the limitations of the immune system. Phage display allows the generation of human antibodies from naive antibody gene libraries when either immunized patients are not available or immunization is not ethically feasible. This technology also allows the construction of immune libraries to select in vivo affinity matured antibodies if immunized patients or animals are available. In this review, we describe the generation of human and human-like antibodies from naive antibody gene libraries and antibodies from immune antibody gene libraries. Furthermore, we give an overview about phage display derived recombinant antibodies against viruses and toxins for diagnostics and therapy.


title: Role of IFN-γ responsiveness in CD8 T cell-mediated viral clearance and demyelination in coronavirus-infected mice

text: Abstract Immunocompetent, but not RAG1−/− mice infected with MHV–JHM develop demyelination. Transferred CD8 T cell-enriched splenocytes reconstitute demyelination, and this ability is dependent on donor IFN-γ. We used IFN-γR1−/− mice to examine the target of IFN-γ in CD8 T cell-mediated demyelination. In IFN-γR1−/−RAG1−/− recipients, demyelination is decreased, but not eliminated, while viral titers are significantly increased when compared to IFN-γR1+/+RAG1−/− recipients. IFN-γR1−/− CD8 T cells retain virus-specific effector function regardless of IFN-γR1 expression. Although IFN-γR1 responsiveness is critical for maximal demyelination, increased levels of infectious virus coupled with adoptive transfer of CD8 T cells may result in myelin destruction independent of IFN-γR1 expression.


Guideline on COVID-19: Ultrasound Machine and Transducer Cleaning


title: 2019 meeting of the global virus network

text: Abstract The Global Virus Network (GVN) was established in 2011 to strengthen research and responses to emerging viral causes of human disease and to prepare against new viral pandemics. There are now 52 GVN Centers of Excellence and 9 Affiliate laboratories in 32 countries. The 11th International GVN meeting was held from June 9–11, 2019 in Barcelona, Spain and was jointly organized with the Spanish Society of Virology. A common theme throughout the meeting was globalization and climate change. This report highlights the recent accomplishments of GVN researchers in several important areas of medical virology, including severe virus epidemics, anticipation and preparedness for changing disease dynamics, host-pathogen interactions, zoonotic virus infections, ethical preparedness for epidemics and pandemics, one health and antivirals.


title: Erythrocytes Reveal Complement Activation in Patients with COVID-19

text: COVID-19, the disease caused by the SARS-CoV-2 virus, can progress to multi-organ failure characterized by respiratory insufficiency, arrhythmias, thromboembolic complications and shock. The mortality of patients hospitalized with COVID-19 is unacceptably high and new strategies are urgently needed to rapidly identify and treat patients at risk for organ failure. Clinical epidemiologic studies demonstrate that vulnerability to organ failure is greatest after viral clearance from the upper airway, which suggests that dysregulation of the host immune response is a critical mediator of clinical deterioration and death. Autopsy and pre-clinical evidence implicate aberrant complement activation in endothelial injury and organ failure. A potential therapeutic strategy warranting investigation is to inhibit complement, with case reports of successful treatment of COVID-19 with inhibitors of complement. However, this approach requires careful balance between the host protective and potential injurious effects of complement activation, and biomarkers to identify the optimal timing and candidates for therapy are lacking. Here we report the presence of complement activation products on circulating erythrocytes from hospitalized COVID-19 patients using flow cytometry. These findings suggest that novel erythrocyte-based diagnostics provide a method to identify patients with dysregulated complement activation.


title: Lipid rafts are involved in SARS-CoV entry into Vero E6 cells

text: Abstract Lipid rafts often serve as an entry site for certain viruses. Here, we report that lipid rafts in Vero E6 cells are involved in the entry of severe acute respiratory syndrome coronavirus (SARS-CoV). Infectivity assay showed the integrity of lipid rafts was required for productive infection of pseudotyped SARS-CoV. Depletion of plasma membrane cholesterol with MβCD relocalized raft-resident marker caveolin-1 as well as SARS-CoV receptor ACE2 to a nonraft environment, but did not significantly change the surface expression of ACE2. MβCD-treatment inhibited infectivity of pseudotyped SARS-CoV by 90%. Biochemical fractionation and confocal imaging confirmed that ACE2 colocalized with raft-resident markers. Furthermore, an ectodomain of SARS-CoV S protein (S1188HA) could associate with lipid rafts after binding to its receptor, and colocalize with raft-resident marker ganglioside GM1. The binding of S1188HA was not affected by depleting plasma membrane cholesterol. Taken together, our results support that lipid rafts serve as an entry port for SARS-CoV.


COVID‐19 and alcohol in Australia: Industry changes and public health impacts


title: First report and phylogenetic analysis of porcine deltacoronavirus in Mexico

text: Porcine deltacoronavirus has caused great economic losses in the swine industry worldwide. In this study, we carried out the first detection, sequencing and characterization of this virus in Mexico. We analysed 885 rectal samples by multiplex RT‐PCR to determine coinfections. In addition, the Spike gene was amplified, sequenced and analysed phylogenetically. We found 85 positive samples for porcine deltacoronavirus, representing 9.6% of the total samples, and we determined that the most frequent coinfection was with porcine epidemic diarrhoea virus (54.1%). Four sequences of Mexican isolates were most closely related to those of the United States. The antigenic regions and the glycosylation site of the strains obtained coincide with those previously reported. This relationship is probably related to the commercial exchange of pigs between the US and Mexico and the geographical proximity of these two countries.


title: Revealing antimicrobial resistance in stormwater with MinION

text: Discharge of urban stormwater containing organic matter, heavy metals and sometime human feces, to the natural aquatic reservoirs without any treatment is not only an environmental problem. It can lead to prevalence of antibiotic resistant bacteria in stormwater systems and transmission of antibiotic resistance genes to the environment. We performed antibiotic resistome identification and virus detection in stormwater samples from Stockholm, using publicly available metagenomic sequencing MinION data. A MinION platform offers low-cost, precise environmental metagenomics analysis. 37 groups of antibiotic resistant bacteria (ARB), 11 resistance types with 26 resistance mechanisms – antibiotic resistance genes (ARGs) giving tolerance to the aminoglycoside, beta-lactams, fosmidomycin, MLS, multidrug and vancomycin were identified using ARGpore pipeline. The majority of the identified bacteria species were related to the natural environment such as soil and were not dangerous to human. Alarmingly, human pathogenic bacteria carrying resistance to antibiotics currently used against them (Bordetella resistant to macrolides and multidrug resistant Propionibacterium avidum) were also found in the samples. Most abundant viruses identified belonged to Caudovirales and Herpesvirales and they were not carrying ARGs. Unlike the virome, resistome and ARB were not unique for stormwater sampling points. This results underline the need for extensive monitoring of the microbial community structure in the urban stormwater systems to assess antimicrobial resistance spread.


title: Porvair highlights its role in supporting the fight against Covid-19

text: Porvair Filtration Group, a specialist in filtration and environmental technologies, has played an important role in supporting the manufacture of components for ventilators and face shields, both of which have been vital during the coronavirus outbreak.


title: Chemical properties of rare earth elements in typical medical waste incinerator ashes in China

text: Abstract Medical waste (MW) ashes from different types of MW incinerators were examined to detect the characteristics and environmental impact of rare earth elements (REEs). The results showed that total REE contents in the ash samples ranged from 10.2 to 78.9mg/kg. REEs in bottom ash were apparently higher than those in fly ash. Average REE contents in the ashes followed the sequence of Ce>La>Nd>Y>Gd>Pr>Sm>Dy>Er>Yb>Ho>Eu>Tb>Lu>Tm. Some of the elements, such as Sm, Dy, Ho, Er, Yb in the ash samples were in normal or nearly normal distribution, but Y, La, Ce, Pr, Nd, Eu, Gd, Tb, Tm, Lu were not normally distributed, indicating some of the ash samples were enriched with these elements. Crust-normalized REE patterns indicated that two types of the MW ashes were obviously enriched with Gd and La. Sequential extraction results showed that REEs in the ash mainly presented as residual fraction, while exchangeable and carbonate fractions were relatively low. DTPA- and EDTA-extraction tests indicated that REEs in the MW ashes were generally in low bioavailability.


title: COVID-19–Related Glomerulopathy: A Report of 2 Cases of Collapsing Focal Segmental Glomerulosclerosis

text: Coronavirus disease 19 (COVID-19), an infectious disease caused by the SARS-CoV-2 virus, has been associated with acute kidney injury (AKI), presumably due to acute tubular injury. However, this does not explain the sometimes severe proteinuria and hematuria often observed. We present 2 African American patients with glomerulopathy demonstrated by kidney biopsy in the setting of AKI and COVID-19 infection. Kidney biopsies showed collapsing variant of focal segmental glomerulosclerosis (FSGS) in addition to acute tubular injury. Both patients were homozygous for APOL1. COVID-19 infection likely caused the interferon surge as a second hit causing podocyte injury leading to collapsing FSGS. APOL1 testing should be strongly considered in African American patients with nephrotic range proteinuria. More data from future kidney biopsies will further elucidate the pathology of kidney injury and glomerular involvement from COVID-19 infections.


title: Studies on Calf Diarrhoea in Mozambique: Prevalence of Bacterial Pathogens

text: The prevalence of diarrhoea in calves was investigated in 8 dairy farms in Mozambique at 4 occasions during 2 consecutive years. A total of 1241 calves up to 6 months of age were reared in the farms, and 63 (5%) of them had signs of diarrhoea. Two farms had an overall higher prevalence (13% and 21%) of diarrhoea. Faecal samples were collected from all diarrhoeal calves (n = 63) and from 330 healthy calves and analysed for Salmonella species, Campylobacter jejuni and enterotoxigenic Escherichia coli (ETEC). Salmonella spp. was isolated in only 2% of all calves. Campylobacter was isolated in 11% of all calves, irrespective of health condition, and was more frequent (25%) in one of the 2 diarrhoeal farms (p = 0.001). 80% of the isolates were identified as C. jejuni. No ETEC strains were detected among the 55 tested strains from diarrhoeal calves, but 22/55 (40%) strains from diarrhoeal calves and 14/88 (16%) strains from healthy calves carried the K99 adhesin (p = 0.001). 6,757 E. coli isolates were typed with a biochemical fingerprinting method (the PhenePlate™) giving the same E. coli diversity in healthy and diarrhoeal calves. Thus it was concluded: i) the overall prevalence of diarrhoea was low, but 2 farms had a higher prevalence that could be due to an outbreak situation, ii) Salmonella did not seem to be associated with diarrhoea, iii) Campylobacter jejuni was common at one of the 2 diarrhoeal farms and iv) ETEC strains were not found, but K99 antigen was more prevalent in E. coli strains from diarrhoeal calves than from healthy, as well as more prevalent in one diarrhoeal farm.


title: Rotavirus infections associated with diarrhoea in calves in Egypt

text: The successful isolation and identification of rotavirus from newborn calves with diarrhoea is reported for the first time in Egypt. From 25 faecal samples taken from diarrhoeic calves, ten virus isolates were found to give cytopathogenic effects on bovine embryonic kidney cells. Three of the isolates were identified as rotavirus using fluorescent antibody staining, serum-neutralization, complement fixation and agar gel precipitation. The complement fixation test revealed the presence of rotavirus antibodies in 18 of 105 serum samples obtained from other calves slaughtered at Cairo abattoir.


title: Porcine epidemic diarrhea virus ORF3 protein causes endoplasmic reticulum stress to facilitate autophagy

text: Abstract Porcine epidemic diarrhea virus (PEDV), the causative agent of PED, is an enveloped, positive-stranded RNA virus in the genus Alphacoronavirus, family Coronaviridae, order Nidovirales. PEDV non-structural accessory protein ORF3 is an ion channel related to viral infectivity and pathogenicity. Our previous study showed that PEDV ORF3 has expression characteristic of aggregation in cytoplasm, but its biological function remains elusive. Thus in this study, we initiated the construction of various vectors to express ORF3, and found ORF3 localized in the cytoplasm in the aggregation manner. Subsequently, confocal microscopy analysis showed that the aggregated ORF3 localized in endoplasmic reticulum (ER) to trigger ER stress response via up-regulation of GRP78 protein expression and activation of PERK-eIF2α signaling pathway. In addition, our results showed that PEDV ORF3 could induce the autophagy through inducing conversion of LC3-I to LC3-II, but couldn’t influence the apoptosis. In contrast, conversion of LC3-I/LC3-II could be significantly inhibited by 4-PBA, an ER stress inhibitor, indicating that ORF3-induced autophagy is dependent on ER stress response. This work not only provides some new findings for the biological function of the PEDV ORF3 protein, but also help us for the further understanding the molecular interaction between PEDV ORF3 protein and cells.


title: The influence of selenium and selenoproteins on immune responses of poultry and pigs

text: Abstract Selenium is an essential nutrient for poultry and pigs, and is important for a number of physiological processes including regulation and function of the immune system. Through its incorporation into selenoproteins, Se is involved in the regulation of oxidative stress, redox mechanisms, and other crucial cellular processes involved in innate and adaptive immune response. This review provides current knowledge on the mechanisms by which selenium can modulate the resilience to infectious diseases, and how this micronutrient can influence the capacity of the bird or the pig to maintain its productivity during an infectious challenge. In relation to the most frequent and economically important infectious diseases in poultry and pig production, the present paper considers the influence of different selenium sources (organic vs. inorganic Se) as well as dietary concentrations on the immune responses of poultry and pigs with major emphasis on the potential beneficial impact on animal resilience to common infectious diseases.


title: Structure of the SARS-CoV NSP12 polymerase bound to NSP7 and NSP8 co-factors

text: Recent history is punctuated by the emergence of highly pathogenic coronaviruses such as SARS- and MERS-CoV into human circulation. Upon infecting host cells, coronaviruses assemble a multi-subunit RNA-synthesis complex of viral non-structural proteins (NSP) responsible for the replication and transcription of the viral genome. Here, we present the 3.1 Å resolution structure of the SARS-CoV NSP12 polymerase bound to its essential co-factors, NSP7 and NSP8, using single particle cryo-electron microscopy. NSP12 possesses an architecture common to all viral polymerases as well as a large N-terminal extension containing a kinase-like fold and is unexpectedly bound by two NSP8 co-factors. This structure illuminates the assembly of the coronavirus core RNA-synthesis machinery, provides key insights into NSP12 polymerase catalysis and fidelity and acts as a template for the design of novel antiviral therapeutics.


title: Epidemic viral gastroenteritis

text: Abstract Epidemic viral gastroenteritis is a significant world wide problem. In developed countries, gastroenteritis accounts for significant morbidity and loss of time from work; in the Third World it is the leading cause of mortality among infants and children. Recent technologic advances have been associated with an explosion of research activity. Two virus groups, the Norwalk-like agents and the rotaviruses, are currently accepted as causative agents of viral gastroenteritis in man. The problem of viral gastroenteritis is reviewed both from a current and a historic perspective.


title: Alignment-free similarity analysis for protein sequences based on fuzzy integral

text: Sequence comparison is an essential part of modern molecular biology research. In this study, we estimated the parameters of Markov chain by considering the frequencies of occurrence of the all possible amino acid pairs from each alignment-free protein sequence. These estimated Markov chain parameters were used to calculate similarity between two protein sequences based on a fuzzy integral algorithm. For validation, our result was compared with both alignment-based (ClustalW) and alignment-free methods on six benchmark datasets. The results indicate that our developed algorithm has a better clustering performance for protein sequence comparison.


title: Lactobacillus Mucosal Vaccine Vectors: Immune Responses against Bacterial and Viral Antigens

text: Lactic acid bacteria (LAB) have been utilized since the 1990s for therapeutic heterologous gene expression. The ability of LAB to elicit an immune response against expressed foreign antigens has led to their exploration as potential mucosal vaccine candidates. LAB vaccine vectors offer many attractive advantages: simple, noninvasive administration (usually oral or intranasal), the acceptance and stability of genetic modifications, relatively low cost, and the highest level of safety possible. Experimentation using LAB of the genus Lactobacillus has become popular in recent years due to their ability to elicit strong systemic and mucosal immune responses. This article reviews Lactobacillus vaccine constructs, including Lactobacillus species, antigen expression, model organisms, and in vivo immune responses, with a primary focus on viral and bacterial antigens.


title: Prediction and biochemical analysis of putative cleavage sites of the 3C-like protease of Middle East respiratory syndrome coronavirus

text: Abstract Coronavirus 3C-like protease (3CLpro) is responsible for the cleavage of coronaviral polyprotein 1a/1ab (pp1a/1ab) to produce the mature non-structural proteins (nsps) of nsp4–16. The nsp5 of the newly emerging Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as 3CLpro and its canonical cleavage sites (between nsps) were predicted based on sequence alignment, but the cleavability of these cleavage sites remains to be experimentally confirmed and putative non-canonical cleavage sites (inside one nsp) within the pp1a/1ab awaits further analysis. Here, we proposed a method for predicting coronaviral 3CLpro cleavage sites which balances the prediction accuracy and false positive outcomes. By applying this method to MERS-CoV, the 11 canonical cleavage sites were readily identified and verified by the biochemical assays. The Michaelis constant of the canonical cleavage sites of MERS-CoV showed that the substrate specificity of MERS-CoV 3CLpro is relatively conserved. Interestingly, nine putative non-canonical cleavage sites were predicted and three of them could be cleaved by MERS-CoV nsp5. These results pave the way for identification and functional characterization of new nsp products of coronaviruses.


title: Microbiological, pathological and histological findings in four Danish pig herds affected by a new neonatal diarrhoea syndrome

text: BACKGROUND: Neonatal diarrhoea is a frequent clinical condition in commercial swine herds, previously regarded to be uncomplicated to treat. However, since 2008 it seems that a new neonatal diarrhoeic syndrome unresponsive to antibiotics and common management practices has emerged. Routine laboratory examinations have not detected any pathogen related to this syndrome. The primary purpose of this study was to evaluate if well-known enteric pathogens could be associated with outbreaks of neonatal diarrhoea, thus question the hypotheses of a new syndrome. Furthermore, we wanted to evaluate macroscopic and microscopic findings associated with these outbreaks and if possible propose a preliminary piglet-level case-definition on syndrome New Neonatal Porcine Diarrhoea syndrome (NNPDS). RESULTS: Four well-managed herds experiencing neonatal diarrhoea with no previously established laboratory conclusion and suspected to suffer from New Neonatal Porcine Diarrhoea Syndrome, were selected. Within these herds, 51 diarrhoeic and 50 non-diarrhoeic piglets at the age of three to seven days were necropsied and subjected to histological and microbiological examination. Faeces were non-haemorrhagic. Neither enterotoxigenic E. coli, Clostridium perfringens type A or C, Clostridium difficile, rotavirus, coronavirus, Cryptosporidium spp, Giardia spp, Cystoisospora suis nor Strongyloides ransomi were associated with diarrhoea in the investigated outbreaks. Macroscopically, the diarrhoeic piglets were characterized by filled stomachs and flaccid intestines without mucosal changes. The predominant histological lesions were villous atrophy in jejunum and ileum. Epithelial lesions in colon were seen in one third of the case piglets. CONCLUSIONS: The results of the study supported the hypothesis that a new neonatal porcine diarrhoea was present in the investigated herds, since no known pathogen(s) or management factors could explain the diarrhoeal outbreaks. Based on the findings in the four herds the following case-definition of NNPDS was suggested: Non-haemorrhagic diarrhoea during the first week of life, without detection of known infectious pathogens, characterized by milk-filled stomachs and flaccid intestines at necropsy.


title: Virtual Spine: A novel, international teleconferencing program developed to increase the accessibility of spine education during the COVID-19 pandemic

text: Background The coronavirus (COVID-19) pandemic effectively ended all major spine educational conferences in the first half of 2020. In response, the authors formed a “virtual” case-based conference series directed at delivering spine education to healthcare providers around the world. We herein share the technical logistics, early participant feedback, and future direction of this initiative. Methods The Virtual Global Spine Conference (VGSC) was created in April 2020 by a multi-institutional team of spinal neurosurgeons and a neuroradiologist. Biweekly virtual meetings were established wherein invited national and international spine care providers would deliver case-based presentations on spine and spine surgery-related conditions via teleconferencing. Promotion was coordinated through social media platforms such as Twitter. Results VGSC recruited over 1000 surgeons, trainees and other specialists, with 50-100 new registrants per week thereafter. An early survey to the participants, with 168 responders, indicated that 92% viewed the content as highly valuable to their practice and 94% would continue participating post-COVID. Participants from the United States (29%), Middle East (16%), and Europe (12%) comprised the majority of the audience. Approximately 52% were neurosurgeons, 18% orthopaedic surgeons, and 6% neuroradiologists. A majority of participants were physicians (55%) and residents/fellows (21%). Conclusion The early success of the VGSC reflects a strong interest in spine education despite the COVID pandemic and social distancing guidelines. There is widespread opinion, backed by our own survey results, that many clinicians and trainees wish to see “virtual” education continue post-COVID.


title: Chitra Ultraviolet-C-Based Facemask Disposal Bin

text: Used face masks are hazardous waste and must be discarded immediately upon removal. Instead of throwing used masks into the disposal bin, disinfecting the masks is essential to break the chain of infection spread. The development of this device was attempted for COVID-19 management, with the following focus: (1) solution which have sufficient science background established so that extensive experimental validation need not be attempted during the lockdown period in India. (2) Provide solution which could be replicated with local resources and minimum material movement. Therefore, in response to the pressing societal demand we designed, prototyped, verified and validated an UV-C-based multipurpose disinfection device. The device after safety and efficacy evaluation as per the Indian Council of Medical Research (ICMR) guidelines for novel COVID-19 solutions, received Central Drugs Control Standard Organization (CDSCO) registration as non-notified medical device for commercialization.


title: Genomic determinants of pathogenicity in SARS-CoV-2 and other human coronaviruses

text: SARS-CoV-2 poses an immediate, major threat to public health across the globe. Here we report an in-depth molecular analysis to reconstruct the evolutionary origins of the enhanced pathogenicity of SARS-CoV-2 and other coronaviruses that are severe human pathogens. Using integrated comparative genomics and machine learning techniques, we identify key genomic features that differentiate SARS-CoV-2 and the viruses behind the two previous deadly coronavirus outbreaks, SARS-CoV and MERS-CoV, from less pathogenic coronaviruses. These features include enhancement of the nuclear localization signals in the nucleocapsid protein and distinct inserts in the spike glycoprotein that appear to be associated with high case fatality rate of these coronaviruses as well as the host switch from animals to humans. The identified features could be crucial elements of coronavirus pathogenicity and possible targets for diagnostics, prognostication and interventions.


title: Infectious causes of necrotizing enterocolitis

text: Necrotizing enterocolitis (NEC) is the most common gastrointestinal emergency among premature infants. Although a large body of research has focused on understanding its pathogenesis, the exact mechanism has not been elucidated. Of particular interest is the potential causative role of infectious culprits in the development of NEC. A variety of reports describe bacterial, viral, and fungal infections occurring in association with NEC; however, no organism has emerged as being definitively involved in NEC pathogenesis. In this review, we summarize the body of research on infectious causes of necrotizing enterocolitis.


title: Diverse activation of microglia by chemokine (C-C motif) ligand 2 overexpression in brain

text: BACKGROUND: The chemokine (C-C motif) ligand 2 (CCL2) is a monocyte chemoattractant protein that mediates macrophage recruitment and migration during peripheral and central nervous system (CNS) inflammation. METHODS: To determine the impact of CCL2 in inflammation in vivo and to elucidate the CCL2-induced polarization of activated brain microglia, we delivered CCL2 into the brains of wild-type mice via recombinant adeno-associated virus serotype 9 (rAAV-9) driven by the chicken β-actin promoter. We measured microglial activation using histological and chemical measurement and recruitment of monocytes using histology and flow cytometry. RESULTS: The overexpression of CCL2 in the CNS induced significant activation of brain resident microglia. CD45 and major histocompatibility complex class II immunoreactivity significantly increased at the sites of CCL2 administration. Histological characterization of the microglial phenotype revealed the elevation of “classically activated” microglial markers, such as calgranulin B and IL-1β, as well as markers associated with “alternative activation” of microglia, including YM1 and arginase 1. The protein expression profile in the hippocampus demonstrated markedly increased levels of IL-6, GM-CSF and eotaxin (CCL-11) in response to CCL2, but no changes in the levels of other cytokines, including TNF-α and IFN-γ. Moreover, real-time PCR analysis confirmed increases in mRNA levels of gene transcripts associated with neuroinflammation following CCL2 overexpression. Finally, we investigated the chemotactic properties of CCL2 in vivo by performing adoptive transfer of bone marrow–derived cells (BMDCs) isolated from donor mice that ubiquitously expressed green fluorescent protein. Flow cytometry and histological analyses indicated that BMDCs extravasated into brain parenchyma and colabeled with microglial markers. CONCLUSION: Taken together, our results suggest that CCL2 strongly activates resident microglia in the brain. Both pro- and anti-inflammatory activation of microglia were prominent, with no bias toward the M1 or M2 phenotype in the activated cells. As expected, CCL2 overexpression actively recruited circulating monocytes into the CNS. Thus, CCL2 expression in mouse brain induces microglial activation and represents an efficient method for recruitment of peripheral macrophages.


title: Foot-and-mouth disease virus utilizes an autophagic pathway during viral replication

text: Abstract Foot-and-mouth disease virus (FMDV) is the type species of the Aphthovirus genus within the Picornaviridae family. Infection of cells with positive-strand RNA viruses results in a rearrangement of intracellular membranes into viral replication complexes. The origin of these membranes remains unknown; however induction of the cellular process of autophagy is beneficial for the replication of poliovirus, suggesting that it might be advantageous for other picornaviruses. By using confocal microscopy we showed in FMDV-infected cells co-localization of non-structural viral proteins 2B, 2C and 3A with LC3 (an autophagosome marker) and viral structural protein VP1 with Atg5 (autophagy-related protein), and LC3 with LAMP-1. Importantly, treatment of FMDV-infected cell with autophagy inducer rapamycin, increased viral yield, and inhibition of autophagosomal pathway by 3-methyladenine or small-interfering RNAs, decreased viral replication. Altogether, these studies strongly suggest that autophagy may play an important role during the replication of FMDV.


title: APOBEC3G cytidine deaminase association with coronavirus nucleocapsid protein

text: Abstract We previously reported that replacing HIV-1 nucleocapsid (NC) domain with SARS-CoV nucleocapsid (N) residues 2–213, 215–421, or 234–421 results in efficient virus-like particle (VLP) production at a level comparable to that of wild-type HIV-1. In this study we demonstrate that these chimeras are capable of packaging large amounts of human APOBEC3G (hA3G), and that an HIV-1 Gag chimera containing the carboxyl-terminal half of human coronavirus 229E (HCoV-229E) N as a substitute for NC is capable of directing VLP assembly and efficiently packaging hA3G. When co-expressed with SARS-CoV N and M (membrane) proteins, hA3G was efficiently incorporated into SARS-CoV VLPs. Data from GST pull-down assays suggest that the N sequence involved in N–hA3G interactions is located between residues 86 and 302. Like HIV-1 NC, the SARS-CoV or HCoV-229E N-associated with hA3G depends on the presence of RNA, with the first linker region essential for hA3G packaging into both HIV-1 and SARS-CoV VLPs. The results raise the possibility that hA3G is capable of associating with different species of viral structural proteins through a potentially common, RNA-mediated mechanism.


title: Severe Acute Respiratory Syndrome Coronavirus 2−Specific Antibody Responses in Coronavirus Disease Patients

text: A new coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has recently emerged to cause a human pandemic. Although molecular diagnostic tests were rapidly developed, serologic assays are still lacking, yet urgently needed. Validated serologic assays are needed for contact tracing, identifying the viral reservoir, and epidemiologic studies. We developed serologic assays for detection of SARS-CoV-2 neutralizing, spike protein–specific, and nucleocapsid-specific antibodies. Using serum samples from patients with PCR-confirmed SARS-CoV-2 infections, other coronaviruses, or other respiratory pathogenic infections, we validated and tested various antigens in different in-house and commercial ELISAs. We demonstrated that most PCR-confirmed SARS-CoV-2–infected persons seroconverted by 2 weeks after disease onset. We found that commercial S1 IgG or IgA ELISAs were of lower specificity, and sensitivity varied between the 2 assays; the IgA ELISA showed higher sensitivity. Overall, the validated assays described can be instrumental for detection of SARS-CoV-2–specific antibodies for diagnostic, seroepidemiologic, and vaccine evaluation studies.


title: COVID-19 and Gastrointestinal Symptoms—A Case Report

text: COVID-19, a new illness secondary to a novel Coronavirus emerged in December 2019 in China. Our early understanding of the clinical features of COVID-19 has been based on case series emerging from the first outbreak in Wuhan. These features included fever, a dry cough, myalgia and dyspnea. Gastrointestinal symptoms were rarely reported as a key feature. We present a case report of a 74-year-old male who presented with symptoms of gastroenteritis and subsequently tested positive for COVID-19. This article aims to highlight an uncommon presentation of COVID-19 and that a high index of suspicion is required for COVID-19 in older people given their greater likelihood of presenting atypically.


title: Coproducing Responses to COVID‐19 with Community‐Based Organizations: Lessons from Zhejiang Province, China

text: Zhejiang Province achieved one of the best records in containing the COVID‐19 pandemic in China, what lessons can the world learn from it? What roles do community‐based organizations play in its success story? Based on more than 100 interviews during and after the outbreak in Zhejiang, this article provides a roadmap of how community‐based organizations were involved in the three distinct stages of Zhejiang's responses to COVID‐19. We recommend that public sector leaders strategically leverage the strengths of community‐based organizations in multiple stages of COVID‐19 responses; incentivize volunteers to participate in epidemic prevention and control; provide data infrastructure and digital tracking platforms; and build trust and long‐term capacity of community‐based organizations. This article is protected by copyright. All rights reserved.


Cerebral hemorrhage related to vein thrombosis in Covid-19 patients in different Italian hospitals: View point for clinical and imaging implications


title: Living and dying in a virtual world

text: A personal narrative is presented which explores the author's experience of being a virtual participant via a conference app in two major family events at the height of the coronavirus disease-2019 (COVID-19) pandemic


title: Rapid in situ hybridization technique for the detection of ribonucleic acids in tissues using radiolabelled and fluorescein-labelled riboprobes

text: Abstract In situhybridization (ISH) is a useful diagnostic and research tool, but is also time consuming. This study was conducted to determine if a rate enhancement hybridization (REH) buffer, developed for membrane hybridization, could be used to decrease hybridization time for ISH. Tissue from swine with an enteric disease produced by a swine coronavirus, transmissible gastroenteritis virus (TGEV), was used as a model to standardize hybridization conditions for a rapid ISH technique. Small intestinal sections from pigs experimentally and naturally infected with TGEV were hybridized for various times at 52°C and 70°C with a radiolabelled or a fluorescein-labelled RNA probe in a standard hybridization or a REH buffer. Viral RNA was detected in intestines from as early as 30 min of hybridization by using both buffers with the radiolabelled probe; however, the signal was stronger with the REH buffer. With the fluorescein-labelled probe, viral RNA was detected in virus-infected cells of the intestines after 30 min of hybridization by using the REH buffer. Signal intensity was greater with the REH buffer than with the standard hybridization buffer when compared at each hybridization time and hybridization temperature using both radiolabelled and fluorescein-labelled probes. With the REH buffer, hybridization signal intensity was greater at 70°C than at 52°C for both probes. The best results were obtained when small intestinal sections were hybridized at 70°C for 2 h using a radiolabelled or a fluorescein-labelled probe diluted in the REH buffer. The fluorescein-labelled RNA probe with REH buffer resulted in a minimal non-specific signal when compared with the radiolabelled probe. These studies demonstrated that the REH buffer can be used to decrease the time of ISH for the detection of viral RNA. This rapid ISH technique should have broad applications in the utilization of probe technology in diagnostics and research for the detection of target ribonucleic acidsin situ


title: Induction of hairy roots by Agrobacterium rhizogenes-mediated transformation of spine gourd (Momordica dioica Roxb. ex. willd) for the assessment of phenolic compounds and biological activities

text: Abstract An efficient protocol for hairy root induction of spine gourd (Momordica dioica) was established using Agrobacterium rhizogenes (KCTC 2703). This study evaluates the phenolic compound production, antioxidant and antimicrobial (antibacterial, antifungal and antiviral) activities of transgenic hairy root cultures in M. dioica . Hairy roots were induced from leaves, petiole, and internodal explants. Molecular analysis of PCR and gene sequencing using specific primers of rolC and aux1 revealed T-DNA integration in the hairy root clones and RT-PCR analysis confirmed the expression of hairy root inducible genes (rolC and aux1). The greatest biomass accumulation of hairy roots on MS liquid medium supplemented with 3% sucrose was observed at 22 days. Ultra-HPLC was used to compare the individual phenolic compound contents of transgenic and non-transgenic roots. Moreover, transgenic hairy roots efficiently produced several phenolic compounds, such as flavonols, hydroxycinnamic acid and hydroxybenzoic acid derivatives. The total phenolic, flavonoid contents and biological (antioxidant, antibacterial, antifungal and antiviral) activities were higher in hairy roots compared to non-transformed roots. These results demonstrate the greater potentiality of M. dioica hairy root cultures for the production of valuable phenolic compounds and for studies of their biological activity.


title: CXCR3 modulates glial accumulation and activation in cuprizone-induced demyelination of the central nervous system

text: BACKGROUND: The functional state of glial cells, like astrocytes and microglia, critically modulates the course of neuroinflammatory and neurodegenerative diseases and can have both detrimental and beneficial effects. Glial cell function is tightly controlled by cellular interactions in which cytokines are important messengers. Recent studies provide evidence that in particular chemokines are important modulators of glial cell function. During the course of CNS diseases like multiple sclerosis or Alzheimer’s disease, and in the corresponding animal models, the chemokines CXCL9 and CXCL10 are abundantly expressed at sites of glial activation, arguing for an important role of these chemokines and their corresponding receptor CXCR3 in glial activation. To clarify the role of this chemokine system in glial cell activation, we characterized the impact of CXCR3 on glial activation in a model of toxic demyelination in which glial activation without a prominent influx of hematogenous cells is prototypical. METHODS: We investigated the impact of CXCR3 on cuprizone-induced demyelination, comparing CXCR3-deficient mice with wild type controls. The clinical course during cuprizone feeding was documented for five weeks and for the subsequent four days withdrawal of the cuprizone diet (5.5 weeks). Glial activation was characterized using histological, histomorphometric and phenotypic analysis. Molecular analysis for (de)myelination and neuroinflammation was applied to characterize the effect of cuprizone on CXCR3-deficient mice and control animals. RESULTS: CXCR3-deficient mice displayed a milder clinical course during cuprizone feeding and a more rapid body weight recovery after offset of diet. In the CNS, CXCR3 deficiency significantly attenuated the accumulation and activation of microglia and astrocytes. Moreover, a deficiency of CXCR3 reduced the expression of the microglial activation markers CD45 and CD11b. Compared to controls, we observed a vast reduction of RNA levels for proinflammatory cytokines and chemokines like Ccl2, Cxcl10, Tnf and Il6 within the CNS of cuprizone-treated mice. Lastly, CXCR3 deficiency had no major effects on the course of demyelination during cuprizone feeding. CONCLUSIONS: The CXCR3 chemokine system is critically involved in the intrinsic glial activation during cuprizone-induced demyelination, which significantly modulates the distribution of glial cells and the local cytokine milieu.


title: Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses

text: A stable culture of primary porcine enterocytes is necessary to study porcine enteric virus replication characteristics. Because the direct cultivation of primary porcine enterocytes is difficult, alternatives have to be considered. As subepithelial myofibroblasts secrete extracellular matrix and growth factors contributing to the attachment, proliferation and differentiation of epithelial cells, co-cultures of primary porcine enterocytes (ileocytes and colonocytes) with myofibroblasts were developed and evaluated for their susceptibility to enteric viruses. First, it was demonstrated that the co-cultured ileocytes and colonocytes were susceptible to an archival rotavirus strain RVA/pig-tc/BEL/RV277/1977/G1P[7] and different other rotavirus genotypes (fecal samples containing G5P[7], G5P[13], G9P[23], G4P[6]). Next, the TGEV Purdue strain infected both ileocytes and colonocytes whereas the Miller strain only infected ileocytes. Last, the PEDV CV777 Vero adapted and non-adapted (fecal suspension) strains could infect co-cultured ileocytes but not colonocytes. The infectivity of the CV777 Vero adapted strain was higher when the cells were cultured without fetal bovine serum and the CV777 fecal suspension only infected the ileocytes cultured without fetal bovine serum. In conclusion, a novel co-culture of porcine enterocytes with myofibroblasts was established, which can be used for the investigation of the replication of enteric viruses.


title: Capacity of serotype 19A and 15B/C Streptococcus pneumoniae isolates for experimental otitis media: implications for the conjugate vaccine

text: Non-vaccine Streptococcus pneumoniae serotypes are increasingly associated with disease. We evaluated isolates of the same sequence type (ST199) but different serotype (15B/C, 19A) for growth in vitro, and pathogenic potential in a chinchilla otitis media model. We also developed a qPCR assay to quantitatively assess each isolate, circumventing the need for selectable markers. In vitro studies showed faster growth of serotype 19A over 15B/C. Both were equally capable of colonization and middle ear infection in this model. Serotype 19A is included in new conjugate vaccine formulations while serotype 15B/C is not. Non-capsular vaccine targets will be important in disease prevention efforts.


title: Yeast surface display for protein engineering and characterization

text: Yeast surface display is being employed to engineer desirable properties into proteins for a broad variety of applications. Labeling with soluble ligands enables rapid and quantitative analysis of yeast-displayed libraries by flow cytometry, while libraries with insoluble or even as-yet-uncharacterized binding targets can be screened through cell-surface selections. In parallel, the utilization of yeast surface display for protein characterization, including in particular the mapping of functional epitopes mediating protein-protein interactions, represents a significant recent advance.


title: Epidemiology of human adenovirus and molecular characterization of human adenovirus 55 in China, 2009–2012

text: BACKGROUND: Human adenovirus 55 (HAdV-55) has caused recent outbreaks of acute respiratory disease (ARD) among adults and military trainees. The active surveillance for HAdV infections was sparse in China, and current knowledge on the HAdV-type distributions and its molecular evolution is lacking. OBJECTIVES: To acquire better understanding on the prevalence and molecular evolution of HAdV-55 strains in China, for an informed strategy for disease control and prevention. POPULATION/METHODS: Nasopharyngeal aspirates were collected from hospitalized children with ARTI in Chongqing during 2009–2012. The genotype of HAdV isolates were determined by sequencing the partial hexon and fiber genes. Whole genome sequences of HAdV-55 were obtained for molecular evolution analysis. RESULTS: About 191 (8·55%) HAdV were detected in 2234 children, including 92 (48·2%) with HAdV-7, 72 (37·7%) with HAdV-3, 6 (3·1%) with HAdV-55, 5 (2·6%) with HAdV-5, 4 (2·1%) with HAdV-1, 1 (0·5%) with HAdV-2, and 11(5·8%) with untyped HAdV. Four of these children developed pneumonia, two of whom were diagnosed with severe pneumonia and/or encephalopathy. HAdV-55 isolates clustered with HAdV-11 sequences based on the hexon gene and clustered with HAdV-14 sequences based on the fiber gene and the whole genome. The overall evolutionary rates of hexon gene, fiber gene, and whole genome of HAdV-55 were estimated at 6·2 × 10(−5) s/s/y, 8·0 × 10(−5) s/s/y, and 1·7 × 10(−5) s/s/y, respectively. CONCLUSIONS: This study suggested HAdV-55 as an emerging infectious disease pathogen has conserved genetic structure and is closely related to each other. Further molecular investigation based on HAdV-55 of wider origin might facilitate understanding its diversity, dissemination, and transmission in China.


title: Oropouche virus entry into HeLa cells involves clathrin and requires endosomal acidification

text: Abstract Oropouche virus (ORO), family Bunyaviridae, is the second most frequent cause of arboviral febrile illness in Brazil. Studies were conducted to understand ORO entry in HeLa cells. Chlorpromazine inhibited early steps of ORO replication cycle, consistent with entry/uncoating. The data indicate that ORO enters HeLa cells by clathrin-coated vesicles, by a mechanism susceptible to endosomal acidification inhibitors. Transmission electron microscopy and immunofluorescence indicated that ORO associates with clathrin-coated pits and can be found in association with late endosomes in a time shorter than 1h.


title: Nurture to nature via COVID-19, a self-regenerating environmental strategy of environment in global context

text: Abstract Coronavirus Disease (COVID-19) has become the largest pandemic that has affected 210 countries. Rolling data indicate that 257,3605 people are infected by the disease, from which 701,838 have recovered and 178,562 have died. No specific medicine or vaccine is available yet to control the disease, hence, social distancing via lockdown is widely adopted as the only preventive measure. Social distancing is observed at different level of strictness in different counties but it almost made the world to stands still. Although scientific articles on this largest social move are scanty, it resulted in benefiting the deteriorated environment to revive back. Many environmental indices such as lowering NO2 and CO2 emissions and reduction in particulate matters in air as a result of less human activities have led to clean air and pollution free water in many countries. Undoubtedly, the world was experiencing pollution in several countries due to mainly human activities including urbanization, industrialization, fossil fuel exhaustion etc. Under such situation a special (natural) a protective measure was awaited to fix environmental issues. Probably, the lockdown is one of the natural effects expected by nature via introduction of COVID-19. It is because, introduction of COVID-19 to nature was an outcome of mutation from two of its pre-existing forms, although, debate on it is still continuing. Viability of CoV-19 virus found to have a lot of correlation with aquatic and terrestrial environmental parameters such as pH, surface type, temperature etc. Air pollution is found to increase the risk of COVID-19 infection, therefore, use of mask and alcohols based standard sterilisers is strongly recommended. However, the self-revival rate of nature shall continue during post-lockdown period and a master plan must be adapted by national and international (mostly political) bodies to revive the Mother Nature completely.


title: Computational Inference of Selection Underlying the Evolution of the Novel Coronavirus, Severe Acute Respiratory Syndrome Coronavirus 2

text: The novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that recently emerged in China is thought to have a bat origin, as its closest known relative (BatCoV RaTG13) was described previously in horseshoe bats. We analyzed the selective events that accompanied the divergence of SARS-CoV-2 from BatCoV RaTG13. To this end, we applied a population genetics-phylogenetics approach, which leverages within-population variation and divergence from an outgroup. Results indicated that most sites in the viral open reading frames (ORFs) evolved under conditions of strong to moderate purifying selection. The most highly constrained sequences corresponded to some nonstructural proteins (nsps) and to the M protein. Conversely, nsp1 and accessory ORFs, particularly ORF8, had a nonnegligible proportion of codons evolving under conditions of very weak purifying selection or close to selective neutrality. Overall, limited evidence of positive selection was detected. The 6 bona fide positively selected sites were located in the N protein, in ORF8, and in nsp1. A signal of positive selection was also detected in the receptor-binding motif (RBM) of the spike protein but most likely resulted from a recombination event that involved the BatCoV RaTG13 sequence. In line with previous data, we suggest that the common ancestor of SARS-CoV-2 and BatCoV RaTG13 encoded/encodes an RBM similar to that observed in SARS-CoV-2 itself and in some pangolin viruses. It is presently unknown whether the common ancestor still exists and, if so, which animals it infects. Our data, however, indicate that divergence of SARS-CoV-2 from BatCoV RaTG13 was accompanied by limited episodes of positive selection, suggesting that the common ancestor of the two viruses was poised for human infection. IMPORTANCE Coronaviruses are dangerous zoonotic pathogens; in the last 2 decades, three coronaviruses have crossed the species barrier and caused human epidemics. One of these is the recently emerged SARS-CoV-2. We investigated how, since its divergence from a closely related bat virus, natural selection shaped the genome of SARS-CoV-2. We found that distinct coding regions in the SARS-CoV-2 genome evolved under conditions of different degrees of constraint and are consequently more or less prone to tolerate amino acid substitutions. In practical terms, the level of constraint provides indications about which proteins/protein regions are better suited as possible targets for the development of antivirals or vaccines. We also detected limited signals of positive selection in three viral ORFs. However, we warn that, in the absence of knowledge about the chain of events that determined the human spillover, these signals should not be necessarily interpreted as evidence of an adaptation to our species.


title: Dynamics and orientation of a cationic antimicrobial peptide in two membrane-mimetic systems

text: Abstract In order to investigate the functional and structural properties of cationic α-helical peptides in two different membranes, we studied the 20-residue peptide maximin H6 in two membrane-mimetic systems by NMR spectroscopy using partially 15N-labeled peptide and paramagnetic relaxation enhancements. Maximin H6, which is found in skin secretions of frogs of the Bombinae family, attacks gram-negative bacteria and acts haemolytically. While the peptide spontaneously folds into similar structures in both neutral dodecylphosphocholine (DPC) and negatively charged sodium dodecyl sulphate (SDS) micelles, its structure is more flexible in SDS as shown by 15N relaxation measurements. In addition, it is bound closer to the surface of the micelle and rotated by ∼70° around its helix axis in the negatively charged membrane surrogate compared to the structure in DPC. This might form the basis for peptide–peptide interactions through a GxxxG motif, which could finally lead to membrane disruption and, thus, preferential attack of negatively charged microbial cell walls.


title: Effects of Sialic Acid Modifications on Virus Binding and Infection

text: Sialic acids (Sias) are abundantly displayed on the surfaces of vertebrate cells, and particularly on all mucosal surfaces. Sias interact with microbes of many types, and are the targets of specific recognition by many different viruses. They may mediate virus binding and infection of cells, or alternatively can act as decoy receptors that bind virions and block virus infection. These nine-carbon backbone monosaccharides naturally occur in many different modified forms, and are attached to underlying glycans through varied linkages, creating significant diversity in the pathogen receptor forms. Here we review the current knowledge regarding the distribution of modified Sias in different vertebrate hosts, tissues, and cells, their effects on viral pathogens where those have been examined, and outline unresolved questions.


title: Gaining insights into human viral diseases through mathematics

text: Mathematical models have been recognized as powerful tools for providing new insights into the understanding of viral dynamics of human diseases at both the population and cellular levels. This article briefly reviews the role of mathematical models and␣their historical precedents for creating new knowledge of the mechanisms of disease pathogenesis, transmission, and control of some human viral infections. Future research in the modelling of infectious diseases will need to rely upon incorporation of the fundamental principles that govern viral dynamics in vivo as well as in the population.


The stress-inducible molecular chaperone GRP78 as potential therapeutic target for Coronavirus infection


title: Serological assays for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), March 2020

text: BACKGROUND: The ongoing coronavirus disease (COVID-19) pandemic has major impacts on health systems, the economy and society. Assessing infection attack rates in the population is critical for estimating disease severity and herd immunity which is needed to calibrate public health interventions. We have previously shown that it is possible to achieve this in real time to impact public health decision making. AIM: Our objective was to develop and evaluate serological assays applicable in large-scale sero-epidemiological studies. METHODS: We developed an ELISA to detect IgG and IgM antibodies to the receptor-binding domain (RBD) of the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We evaluated its sensitivity and specificity in combination with confirmatory microneutralisation (MN) and 90% plaque reduction neutralisation tests (PRNT(90)) in 51 sera from 24 patients with virologically confirmed COVID-19 and in age-stratified sera from 200 healthy controls. RESULTS: IgG and IgM RBD ELISA, MN and PRNT(90) were reliably positive after 29 days from illness onset with no detectable cross-reactivity in age-stratified controls. We found that PRNT(90) tests were more sensitive in detecting antibody than MN tests carried out with the conventional 100 tissue culture infectious dose challenge. Heparinised plasma appeared to reduce the infectivity of the virus challenge dose and may confound interpretation of neutralisation test. CONCLUSION: Using IgG ELISA based on the RBD of the spike protein to screen sera for SARS-CoV-2 antibody, followed by confirmation using PRNT(90), is a valid approach for large-scale sero-epidemiology studies.


Phenotypic Expressions of Childhood Wheezing and Asthma: Implications for Therapy


title: Whole genome analysis of Japanese bovine toroviruses reveals natural recombination between porcine and bovine toroviruses

text: Abstract Bovine toroviruses (BToVs), belong to the subfamily Toroviridae within the family Coronaviridae, and are pathogens, causing enteric disease in cattle. In Japan, BToVs are distributed throughout the country and cause gastrointestinal infection of calves and cows. In the present study, complete genome sequences of two Japanese BToVs and partial genome sequences of two Japanese BToVs and one porcine torovirus (PToV) from distant regions in Japan were determined and genetic analyses were performed. Pairwise nucleotide comparison and phylogenetic analyses revealed that Japanese BToVs shared high identity with each other and showed high similarities with BToV Breda1 strain in S, M, and HE coding regions. Japanese BToVs showed high similarities with porcine toroviruses in ORF1a, ORF1b, and N coding regions and the 5′ and 3′ untranslated regions, suggestive of a natural recombination event. Recombination analyses mapped the putative recombinant breakpoints to the 3′ ends of the ORF1b and HE regions. These findings suggest that the interspecies recombinant nature of Japanese BToVs resulted in a closer relationship between BToV Breda1 and PToVs.


title: Genome Sequencing of a Severe Acute Respiratory Syndrome Coronavirus 2 Isolate Obtained from a South African Patient with Coronavirus Disease 2019

text: As a contribution to the global efforts to track and trace the ongoing coronavirus pandemic, here we present the sequence, phylogenetic analysis, and modeling of nonsynonymous mutations for a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome that was detected in a South African patient with coronavirus disease 2019 (COVID-19).


title: Antiviral activity in vitro of two preparations of the herbal medicinal product Sinupret® against viruses causing respiratory infections

text: Abstract Sinupret®, a herbal medicinal product made from Gentian root, Primula flower, Elder flower, Sorrel herb, and Verbena herb is frequently used in the treatment of acute and chronic rhinosinusitis and respiratory viral infections such as common cold. To date little is known about its potential antiviral activity. Therefore experiments have been performed to measure the antiviral activity of Sinupret® oral drops (hereinafter referred to as “oral drops”) and Sinupret® dry extract (hereinafter referred to as “dry extract”), in vitro against a broad panel of both enveloped and non-enveloped human pathogenic RNA and DNA viruses known to cause infections of the upper respiratory tract: influenza A, Chile 1/83 (H1N1) virus (FluA), Porcine Influenza A/California/07/2009 (H1N1) virus (pFluA), parainfluenza type 3 virus (Para 3), respiratory syncytial virus, strain Long (RSV), human rhinovirus B subtype 14 (HRV 14), coxsackievirus subtype A9 (CA9), and adenovirus C subtype 5 (Adeno 5). Concentration-dependent antiviral activity (EC50 between 13.8 and 124.8μg/ml) of Sinupret® was observed against RNA as well as DNA viruses independent of a viral envelope. Remarkable antiviral activity was shown against Adeno 5, HRV 14 and RSV in which dry extract was significantly superior to oral drops. This could be ascertained with different assays as plaque-reduction assays in plaque forming units (PFU), the analyses of a cytopathogenic effect (CPE) and with enzyme immunoassays (ELISA) to determine the amount of newly synthesised virus. Our results demonstrate that Sinupret® shows a broad spectrum of antiviral activity in vitro against viruses commonly known to cause respiratory infections.


title: Chest imaging of H7N9 subtype of human avian influenza

text: Abstract Background Human infection with avian influenza A H7N9 virus is an acute respiratory infectious disease, which usually causes severe pneumonia with a high mortality. Chest radiographs and Computed Tomography (CT) are principal radiological modalities to assess the lung abnormalities. Objectives The goal of this study was to investigate the chest images characteristic of H7N9 subtype of human avian influenza. Materials and methods The clinical and imaging data of 11 cases diagnosed as H7N9 subtype of human avian influenza were collected from 4 cities in the southern region of the Yangtze River, China. The chest imaging manifestations were analyzed by the assigned expert group. The analyzed cases include 7 males and 4 females aged from 20 to 84 years, with a mean of 55.6 years. The clinical symptoms were mainly fever (100%, 11/11) and cough (72.7%, 8/11). Results Segmental or lobar ground-glass opacity (GGO) or consolidation was shown in 8 cases (72.7% or 8/11). Air bronchogram was found in 7 cases (63.6% or 7/11). The lesions developed into multiple or diffuse in both lungs rapidly at the progressive stage. The reticulation shadows were shown after some lesions absorbed at the stable stage. Conclusions The characteristic imaging demonstrations of H7N9 subtype of human avian influenza are segmental or lobar exudative lesions at lungs at the initial stage, which rapidly progress into bilateral distribution at lungs at the progressive stage.


title: Detection of feline Coronavirus in effusions of cats with and without feline infectious peritonitis using loop-mediated isothermal amplification

text: Abstract Feline infectious peritonitis (FIP) is a fatal disease in cats worldwide. The aim of this study was to test two commercially available reaction mixtures in a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to detect feline Coronavirus (FCoV) in body cavity effusions of cats with and without FIP, in order to minimize the time from sampling to obtaining results. RNA was extracted from body cavity effusion samples of 71 cats, including 34 samples from cats with a definitive diagnosis of FIP, and 37 samples of control cats with similar clinical signs but other confirmed diseases. Two reaction mixtures (Isothermal Mastermix, OptiGene Ltd.and PCRun™ Molecular Detection Mix, Biogal) were tested using the same primers, which were designed to bind to a conserved region of the FCoV membrane protein gene. Both assays were conducted under isothermal conditions (61 °C–62 °C). Using the Isothermal Mastermix of OptiGene Ltd., amplification times ranged from 4 and 39 min with a sensitivity of 35.3% and a specificity of 94.6% for the reported sample group. Using the PCRun™ Molecular Detection Mix of Biogal, amplification times ranged from 18 to 77 min with a sensitivity of 58.8% and a specificity of 97.3%. Although the RT-LAMP assay is less sensitive than real time reverse transcription PCR (RT-PCR), it can be performed without the need of expensive equipment and with less hands-on time. Further modifications of primers might lead to a suitable in-house test and accelerate the diagnosis of FIP.


title: Transient transport model of particles resulting from high momentum respiratory activities: Inter-personal exposure

text: Abstract In this work, a transient mathematical multi-region zonal transport model of particle behavior resulting from high momentum respiratory activities (HMRA) is developed focusing on the transient inter-personal exposure (IPE) in indoor spaces ventilated by displacement ventilation (DV) systems. The developed model was validated by experimentation and by published empirical data. Three stages are identified with respect to time for the variation of the IPE: a first stage dominated by the propagation and decay of the exhaled jet, a particles' redistribution stage, and a particles' removal stage. The inhaled dose is affected by the DV flow rate, cough velocity, particle diameter and distance between the occupants. The DV system with a flow rate of 100 L/s reduced significantly the inhaled dose during particle redistribution and removal stages decreasing the total inhaled dose by 83% compared to a flow rate of 50 L/s. IPE is higher when particle diameter is increased from 1 to 20 μm due to the opposition of particle removal by the upward DV. A comparison between steady and transient modeling of the IPE showed that steady modeling captures the physics affecting particle spread due to HMRA but it over-predicts the inhaled dose. It is found that for a DV flow rate of 100 L/s and a cough velocity of 22 m/s during 1 s, and 1 μm particles, the minimum required distance between the occupants for a threshold inhaled dose of 10−5 kg is nearly 0.5 m by transient modeling while it is 2.15 m by steady state modeling.