title: Identification of C-geranylated flavonoids from Paulownia catalpifolia Gong Tong fruits by HPLC-DAD-ESI-MS/MS and their anti-aging effects on 2BS cells induced by H2O2

text: Abstract The fruits of Paulownia catalpifolia Gong Tong are used as a Chinese folk herbal medicine for the treatment of enteritis, tonsillitis, bronchitis, and dysentery, etc. Our previous study has identified new C-geranylated flavanones with obvious anti-proliferative effects in lung cancer A549 cells. In the present study, a new C-geranylated flavone, paucatalinone C (1) and five known C-geranylated flavanones (2–6) were isolated. In addition, a total of 34 C-geranylated flavonoids were detected by HPLC-DAD-ESI-MS/MS coupling techniques from the CH2Cl2 extract of P. catalpifolia. Futhermore, anti-aging effects of isolated compounds were evaluated in vitro with premature senescent 2BS cells induced by H2O2. Phytochemical results indicated that P. catalpifolia was a natural resource of abundant C-geranylated flavonoids. Diplacone (3) and paucatalinone A (5) were the potent anti-aging agents in the premature senescent 2BS cells induced by H2O2 and the C-geranyl substituent may be an important factor because of its lipophilic character.


title: Co‐infection with COVID‐19 and influenza A virus in two died patients with acute respiratory syndrome, Bojnurd, Iran

text: In some recently published papers researchers reported "Co-infection with SARS-CoV-2 and Influenza Virus"1-3 . They described the development of clinical symptoms of pneumoniae in patients and discussed about the importance of other respiratory viruses that cause symptoms similar to COVID-19. This article is protected by copyright. All rights reserved.


Virtual Assessment and Management in Foot and Ankle Surgery During the COVID-19 Pandemic: An Irish Experience


Tratamiento del catarro. Evidencia científica


Impact on colorectal cancer in COVID‐19 pandemic


title: Quantitative temporal‐spatial distribution of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) in post‐mortem tissues

text: Few post‐mortem studies have been performed on patients who have died from severe acute respiratory syndrome (SARS). No studies have examined how the SARS‐associated coronavirus (SARS‐CoV) loads in different organs with respect to time, post‐mortem. The aim of this study was to determine the quantitative temporal‐spatial distribution of SARS‐CoV in the post‐mortem tissue samples of seven patients. Quantitation of a house‐keeping gene, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) was undertaken to standardize the amount of tissue tested. SARS‐CoV viral load and SARS‐CoV/GAPDH RNA ratio for each organ type were related to four time durations: onset of illness to death, death to post‐mortem tissue sampling, and total durations of treatment with ribavirin and hydrocortisone. The SARS‐CoV/GAPDH RNA ratio remained relatively stable in most organ tissue types for all these time durations. The ratio reached the highest value of equal to or greater than one for lung and small bowel, whereas those for heart, liver, spleen, and kidney were always less than one. It is concluded that SARS‐CoV viral loads in these organs remain relatively stable, post‐mortem. This quantitative assessment further supports SARS‐CoV has a specific tropism for the human respiratory and gastrointestinal tracts, which may be related to the density of SARS‐CoV receptors. J. Med. Virol. 79:1245–1253, 2007. © Wiley‐Liss, Inc.


title: Enzyme-linked immunosorbent assay for the detection of canine coronavirus and its antibody in dogs

text: Abstract Two methods of enzyme-linked immunosorbent assay (ELISA) were developed for the diagnosis of canine coronavirus (CCV) infection in dogs. One ELISA, in which CCV-infected CRFK cell lysate is used as antigen, is for the detection and titration of antibody against CCV, and the other ELISA uses the double antibody sandwich method for the detection of CCV antigen. The first ELISA procedure demonstrated antibody responses in dogs inoculated with CCV, as did the virus neutralization test; the second ELISA detected specific CCV antigen in feces and organ homogenates of inoculated dogs.


title: Evolution of Mouse Hepatitis Virus (MHV) during Chronic Infection: Quasispecies Nature of the Persisting MHV RNA

text: Abstract Coronavirus infection of mice has been used extensively as a model for the study of acute encephalitis and chronic demyelination. To examine the evolution of coronavirus RNA during chronic demyelinating infection, we isolated RNA from intracerebrally inoculated mice at 4, 6, 8, 13, 20, and 42 days postinfection and used reverse transcription-polymerase chain reaction amplification methods (RT-PCR) to detect viral sequences. RNA sequences from two viral structural genes, the spike gene and the nucleocapsid gene, were detected throughout the chronic infection. In contrast, infectious virus was not detectable from brain homongenates beyond 13 days postinfection. These results indicate that coronavirus RNA persists in the brain at times when infectious virus is not detected. To determine if genetic changes were occurring during viral replication in the host, we cloned and sequenced the RT-PCR products from the spike and nucleocapsid regions and analyzed the sequences for mutations. Sequencing of the cloned products revealed that a variety of mutant forms of viral RNA persisted in the CNS, including point mutants, deletion mutants, and termination mutants. The mutations accumulated during persistent infection in both the spike and the nucleocapsid sequences, with greater than 65% of the mutations encoding amino acid changes. These results show that a diverse population or quasispecies consisting of mutant and deletion variant viral RNAs (which may not be capable of producing infectious virus particles) persists in the central nervous system of mice during chronic demyelinating infection. The implications of these results for the role of persistent viral genetic information in the pathogenesis of chronic demyelination are discussed.


title: Chest radiograph screening for severe acute respiratory syndrome in the ED

text: Abstract The purpose of the study was to evaluate the use of chest radiography for the screening of severe acute respiratory syndrome (SARS). We retrospectively analyzed all patients who attended an Emergency Department SARS screening clinic during the outbreak in Hong Kong, from March 10 to June 5, 2003. Patients with clinical and epidemiologic suspicion of SARS were evaluated by serial chest radiography. All radiographs were reported by consensus from 2 radiologists, blinded to the clinical records. The prevalence of SARS was 13.3% among 1328 patients included. The initial radiograph had sensitivity 50.3%, specificity 95.0%, positive likelihood ratio 10.06, negative likelihood ratio 0.52, positive predictive value 61.5%, and negative predictive value 92.3% for diagnosing SARS. Serial chest radiography had sensitivity 94.4%, specificity 93.9%, positive likelihood ratio 15.48, negative likelihood ratio 0.06, positive predictive value 71.4%, and negative predictive value 99.0%. The initial chest radiograph has poor sensitivity, and serial radiographs are required to rule out SARS.


title: Epidemiologic and clinical characteristics of 10 children with coronavirus disease 2019 in Changsha, China

text: Abstract Background The outbreak of a new coronavirus, first reported in Wuhan, China, is spreading around the world. Information on the characteristics of children with Coronavirus Disease 2019 (COVID-19) is limited. Methods In this retrospective study, we recruited 10 children infected with SARS-COV-2 from January 27 to March 10, 2020, in Changsha, China. We report the epidemiological, clinical, laboratory, and high-resolution CT findings for these children. Qualitative descriptive analysis was used to describe the key results. Results Ten children were included. Three were male and seven were female. Three were from Wuhan, Hubei Province, and seven were from Changsha. All had a history of close contact with adults with COVID-19 before the onset of disease. Clinical manifestations included fever in four cases, respiratory symptoms in three cases, febrile convulsions in one case, vomiting in one case, abdominal pain in one case, and asymptomatic infection in two cases. All the children tested positive for nucleic acid in throat swabs at admission. Stool swabs of three cases were positive for nucleic acid after several days of fever. In nine children, blood routine results were normal, whereas in one case the white blood cell count was elevated. In four cases, CT findings of the lungs showed light ground-glass opacities, one case showed changes similar to bronchopneumonia, and the remaining cases were normal. All were treated with symptomatic support without complications. Conclusion Our findings indicate that intrafamily transmission may be the main form of transmission of COVID-19 in children, and persistent intestinal excretion of virus is another characteristic among children. The results of stool swab tests should be considered for discharge and release from isolation.


title: Tollwut (Rabies, Lyssa)

text: Zusammenfassung Die Tollwut gehört in Deutschland inzwischen zu den sogenannten vergessenen Krankheiten, obwohl weltweit jährlich mehr als 55 000 Fälle auftreten und mehr als 10 Millionen Post-Expositions-Impfungen durchgeführt werden. Das die Krankheit auslösende Agens ist ein umhülltes Virus. Nach Biss einer Fledermaus, oder meist eines Fuchses in Mitteleuropa, eines Hundes in Afrika, Asien und Südamerika oder eines anderen Tieres wird das Virus in Muskelzellen aufgenommen, vermehrt sich und bekommt einige Zeit später Zugang zu Nervenzellen. Das Virus lagert sich z.B. an den Acteycholin-Rezeptor an und wird im Axon zum Gehirn transportiert und vermehrt sich intrazellulär. Infizierte Nervenzellen ändern ihre Funktion. Die Inkubationszeit bis zum Auftreten von Symptomen beträgt etwa 2 Wochen bis Monate. Symptome sind Enzephalitis oder Paralyse, gefolgt von Hypersalivation, Hydrophobie, Koma und Tod. Die Krankheit beim Menschen verläuft bei Nicht-Geimpften ohne immunologische Intervention, d.h. ohne die Post-Expositions-Impfung, immer tödlich. Eine antivirale Therapie steht nicht zur Verfügung. Die Krankheit Tollwut kann durch präventive und bei Verletzung an der unteren Extremität durch postexpositionelle Impfung verhindert werden. Natürliche Wirte sind Kleinnager, Fledermäuse und Karnivoren; insofern ist Tollwut eine Zoonose und das Virus kann nicht ausgerottet werden. Abstract Rabies is one of the so-called lost diseases in Germany despite every year more than 55.000 cases occur worldwide and more than 10 million post-exposition immunisations are being accomplished. The disease is caused by a virus. After a bite by a bat or, more often, a fox in Central Europe, or a dog in Africa, Asia or South America or any other animal the virus is being incorporated in muscle cells, multiplies and after a while gets access to nerve cells. The virus attaches e.g. to acetylcholine receptors, is transported via the axon to the brain where it reproduces intracellularly. Infected nerve cells change their function. Incubation period up to the appearance of any signs and symptoms is approximately 2 weeks to months. Signs and symptoms are encephalitis or paralysis followed by hypersalivation, hydrophobia, coma and death. In people who have no vaccine protection and do not receive post-exposure vaccination the disease is lethal throughout. There is no antiviral therapy available. Rabies can be prevented by pre-emptive immunisation or after injuries of the lower limb by post-exposure prophylaxis. Natural hosts are small rodents, bats, and carnivores; in this respect rabies is a zoonotic disease and the virus can not be extinguished.


Même sans gravité, un Covid-19 conférerait une immunité


title: Porcine reproductive–respiratory syndrome virus infection predisposes pigs for respiratory signs upon exposure to bacterial lipopolysaccharide

text: Abstract This study examined whether an infection with porcine reproductive and respiratory syndrome virus (PRRSV) potentiates respiratory signs upon exposure to bacterial lipopolysaccharides (LPS). Five-week-old conventional pigs were inoculated intratracheally with the Lelystad strain of PRRSV and received 5 days later one or two intratracheal LPS administrations. The necessary controls were included. After LPS administration, pigs were intensively monitored for clinical signs. Additionally, some pigs were euthanatized after a second LPS administration for broncho-alveolar cell analysis and virological examinations of the lungs. Broncho-alveolar lavage (BAL) cells were counted and differentiated. Lung suspensions and BAL fluids were titrated for PRRSV. Exposure of pigs to PRRSV only resulted in a fever for time periods ranging from 1 to 5 days and slight respiratory signs. Exposure of pigs to LPS only resulted in general signs, characterized by fever and depression, but respiratory signs were slight or absent. PRRSV–LPS exposed pigs, on the other hand, developed severe respiratory signs upon LPS exposure, characterized by tachypnoea, abdominal breathing and dyspnoea. Besides respiratory signs, these pigs also showed enhanced general signs, such as fever and depression. Lung neutrophil infiltration was similar in non-infected and PRRSV-infected pigs upon LPS exposure. PRRSV quantities were similar in lungs and BAL fluids of pigs infected with PRRSV only and PRRSV–LPS exposed pigs. These data show a clear synergism between PRRSV and LPS in the induction of respiratory signs in conventional pigs. The synergism was observed in 87% of the pigs. So, it can be considered as reproducible and may be used to test the efficacy of preventive and therapeutic measures.


title: Advancing Human Health in the Decade Ahead: Pregnancy as a Key Window for Discovery A Burroughs Wellcome Fund Pregnancy Think-Tank

text: Abstract Recent revolutionary advances at the intersection of medicine, omics, data sciences, computing, epidemiology and related technologies inspire us to ponder their impact on health. Their potential impact is particularly germane to the biology of pregnancy and perinatal medicine, where limited improvement in health outcomes for women and children has remained a global challenge. We assembled a group of experts to establish a Pregnancy Think Tank to discuss a broad spectrum of major gestational disorders and adverse pregnancy outcomes that impact maternal-infant lifelong health and that should serve as targets for leveraging the many recent advances. This report reflects avenues for future impact that hold great potential in three major areas: developmental genomics, including the application of methodologies designed to bridge genotypes, physiology and diseases addressing vexing questions in early human development; gestational physiology, from immune tolerance to growth and the timing of parturition; and personalized and population medicine, focusing on amalgamating health record data and deep phenotypes to create broad knowledge that can be integrated into health care systems and drive discovery to address pregnancy related disease and promote general health. We propose a series of questions reflecting development, systems biology, diseases, clinical approaches and tools, and population health and a call for scientific action. Clearly, transdisciplinary science must advance and accelerate in order to address adverse pregnancy outcomes. Disciplines not traditionally involved in the reproductive sciences, including computer science, engineering, mathematics, and pharmacology should be engaged at the study design phase to optimize the information gathered as well as to identify and further evaluate potentially actionable therapeutic targets. Information sources should include non-invasive, personalized sensors and monitors, alongside instructive “liquid biopsies” for non-invasive pregnancy assessment. Future research should also address the diversity of human cohorts in terms of geography, racial and ethnic distributions, as well as social and health disparities. Modern technologies, both for data-gathering and for data-analyzing, make this possible at a scale that was previously unachievable. Finally, the psychosocial and economic environment in which pregnancy takes place must be considered to promote the health and wellness of communities worldwide.


Respirology year‐in‐review 2006: Clinical science


title: Gestion de l’épidémie de la COVID-19 par les établissements publics de santé – Analyse de la Fédération Hospitalière de France

text: Résumé Le but de cet article est de détailler les mesures prises dans les établissements publics pour faire face à l’épidémie de COVID-19. Sont détaillées la stratégie initiale, l’évolution organisationnelle vers le « tout-COVID », la coordination entre les différents intervenants ainsi que la stratégie de maintien de la continuité des soins. L’expérience au Québec est aussi prise en exemple. Enfin la sortie de crise doit être anticipée dès cette phase d’épidémie. Summary The objective of this article is to detail the measures taken in public institutions to cope with the COVID-19 epidemic. It details the initial strategy, organizational evolution towards "all-COVID", coordination between the various stakeholders and the strategy for maintaining continuity of care. The Quebec experience is also used as an example. Finally, an exit strategy must be anticipated at this phase of the epidemic.


title: The Clinics of HHV‐6 infection in COVID‐19 pandemic: Pityriasis rosea and Kawasaki disease

text: A new type of coronavirus family (SARS‐CoV‐2), which can be found in humans and animals, with many varieties and clinical symptoms, was first seen in Wuhan, China in late 2019, under the name novel Coronavirus Disease 2019 (COVID‐19). In the literature, cutaneous symptoms related to the disease are generally emphasized. However, it is not yet known whether this new SARS‐CoV‐2 virus, which has entered our lives, plays a role in the etiopathogenesis of dermatological diseases. The patients who were admitted to the dermatology outpatient clinic between 1 April and May 15, 2019, and on 1 April and May 15, 2020 were retrospectively analyzed by searching the hospital automation system and patient files. The reason for the same months to be included in the study was to exclude seasonal effects on the diseases. After pandemic, the number of patients with Pityriasis rosea and Kawasaki disease increased significantly in patients who applied to the dermatology outpatient clinic. Our study is the first study showing pityriasis rosea increase during the pandemic period. We think that this increase is related to HHV‐6 reactivation. Herein, we wanted to draw attention to two diseases in which Human Herpes 6 (HHV‐6) was accused in etiopathogenesis: Kawasaki disease and Pityriasis rosea. This article is protected by copyright. All rights reserved.


title: Human bocavirus and human metapneumovirus in hospitalized children with lower respiratory tract illness in Changsha, China

text: BACKGROUND: Lower respiratory tract illness is a major cause of morbidity and mortality in children worldwide, however, information about the epidemiological and clinical characteristics of LRTIs caused by HMPV and HBoV in China is limited. OBJECTIVES: Human bocavirus (HBoV) and human metapneumovirus (HMPV) are two important viruses for children with lower respiratory tract infections (LRTI). We aimed to assay the correlation between viral load and clinical characteristics of HBoV and HMPV with LRTI in Changsha, China. METHODS: Nasopharyngeal aspirates (NPAs) from children with LRTI were collected. Real‐time PCR was used to screen HBoV and HMPV. Analyses were performed using SPSS 16.0 software. RESULTS: Pneumonia was the most frequent diagnosis. There was no significant difference between HBoV‐ and HMPV‐positive patients in age (P = .506) or hospitalization duration (P = .280); 24.1% and 18.2% were positive for HBoV and HMPV. HBoV infections peaked in summer (32.2%), and HMPV infections peaked in winter (28.9%). The HBoV‐positive patients had a shorter hospitalization duration than the HBoV‐negative patients (P = .021), and the HMPV‐positive patients had a higher prevalence of fever than the HMPV‐negative patients (P = .002). The HBoV viral load was significantly higher among patients aged <1 year (P = .006). The mean HBoV and HMPV viral loads were not significantly different between patients with single infections and coinfections. Patients infected with HBoV only were older than those coinfected with HBoV and other respiratory viruses (P = .005). No significant difference was found in the clinical characteristics of patients infected with HMPV only and those coinfected with HMPV and other respiratory viruses. CONCLUSION: Pneumonia was the most frequent diagnosis caused by HBoV and HMPV. Neither HBoV nor HMPV viral load was correlated with disease severity.


8th Course of the European School of Neuroimmunology


title: 47 The Aircraft Cabin Environment

text: Abstract Travel by air is a safe means of transport, but from the physiologic point of view it is a means of putting people at risk as well as a potential means of spreading infectious disease. Atmospheric pressure falls with altitude and above about 10,000 feet, blood desaturation leads to hypoxia. The aircraft cabin is pressurized to maintain an effective altitude below 8000 feet, which provides adequate protection for healthy travelers. Those individuals with cardiopulmonary impairment or other conditions affecting fitness to travel should seek advice from the airline in consultation with their medical provider. People are the primary source of microorganisms in aircraft cabins and are the most important reservoirs of infectious agents on aircraft. Recirculated cabin air is filtered to remove microorganisms, but this does not prevent person-to-person spread of infectious disease in vulnerable people. The World Health Organization and Centers for Disease Control and Prevention produce guidelines; and people with febrile illnesses should postpone air travel.


title: Human viruses: discovery and emergence

text: There are 219 virus species that are known to be able to infect humans. The first of these to be discovered was yellow fever virus in 1901, and three to four new species are still being found every year. Extrapolation of the discovery curve suggests that there is still a substantial pool of undiscovered human virus species, although an apparent slow-down in the rate of discovery of species from different families may indicate bounds to the potential range of diversity. More than two-thirds of human viruses can also infect non-human hosts, mainly mammals, and sometimes birds. Many specialist human viruses also have mammalian or avian origins. Indeed, a substantial proportion of mammalian viruses may be capable of crossing the species barrier into humans, although only around half of these are capable of being transmitted by humans and around half again of transmitting well enough to cause major outbreaks. A few possible predictors of species jumps can be identified, including the use of phylogenetically conserved cell receptors. It seems almost inevitable that new human viruses will continue to emerge, mainly from other mammals and birds, for the foreseeable future. For this reason, an effective global surveillance system for novel viruses is needed.


title: Differential effect of cholesterol on type I and II feline coronavirus infection

text: Feline infectious peritonitis (FIP) is a fatal disease of domestic and wild felidae that is caused by feline coronavirus (FCoV). FCoV has been classified into types I and II. Since type I FCoV infection is dominant in the field, it is necessary to develop antiviral agents and vaccines against type I FCoV infection. However, few studies have been conducted on type I FCoV. Here, we compare the effects of cholesterol on types I and II FCoV infections. When cells were treated methyl-β-cyclodextrin (MβCD) and inoculated with type I FCoV, the infection rate decreased significantly, and the addition of exogenous cholesterol to MβCD-treated cells resulted in the recovery of the infectivity of type I FCoV. Furthermore, exogenous cholesterol increased the infectivity of type I FCoV. In contrast, the addition of MβCD and exogenous cholesterol had little effect on the efficiency of type II FCoV infection. These results strongly suggest that the dependence of infection by types I and II FCoV on cholesterol differs.


title: Inanimate surfaces as potential source of 2019-nCoV spread and their disinfection with biocidal agents

text: The WHO has declared COVID-19 illness a global health concern which is caused by 2019-nCoV, causing severe respiratory tract infections in humans. Transmissibility among individual to individual have been reported through droplets and probably also via contaminated surfaces and hands. Human coronaviruses can persist on inanimate surfaces such as plastic, glass, fibers and metals up to nine days. 2019-nCoV remains infectious in air for 3 h and on inanimate surfaces such as cardboard, copper, plastic and steel up to 24, 4, 72 and 48 h respectively. Disinfectant activity of various biocidal agents against coronaviruses like ethanol (62–71%), sodium hypochlorite (0.1%) and hydrogen peroxide (0.5%) can be regarded effective against 2019-nCoV as well. As no vaccine and antiviral therapies have been discovered for 2019-nCoV, prevention of further spread will viable option to control the ongoing and future outbreaks.


title: Spatial Analysis on Hepatitis C Virus Infection in Mainland China: From 2005 to 2011

text: BACKGROUND: The burden of Hepatitis C virus (HCV) has become more and more considerable in China. A macroscopic spatial analysis of HCV infection that can provide scientific information for further intervention and disease control is lacking. METHODS: All geo-referenced HCV cases that had been recorded by the China Information System for Disease Control and Prevention (CISDCP) during 2005–2011 were included in the study. In order to learn about the changes of demographic characteristics and geographic distribution, trend test and spatial analysis were conducted to reflect the changing pattern of HCV infection. RESULTS: Over 770,000 identified HCV infection cases had specific geographic information during the study period (2005–2011). Ratios of gender (Male/Female, Z-value = −18.53, P<0.001), age group (≤30 years old/≥31 years old, Z-value = −51.03, P<0.001) and diagnosis type (Clinical diagnosis/Laboratory diagnosis, Z-value = −130.47, P<0.001) declined. HCV infection was not distributed randomly. Provinces Henan, Guangdong, Guangxi, Xinjiang, and Jilin reported more than 40,000 HCV infections during 2005 to 2011, accounting for 43.91% of all cases. The strength of cluster of disease was increasing in China during the study period. Overall, 11 provinces had once been detected as hotspots during 7 years, most of which were located in the central or border parts of China. Tibet, Qinghai, Jiangxi were the regions that had coldspots. CONCLUSIONS: The number of clustering of HCV infection among older adults increased in recent years. Specific interventions and prevention programs targeting at main HCV epidemic areas are urgently in need in mainland China.


title: Induction of class I antigen processing components in oligodendroglia and microglia during viral encephalomyelitis

text: Glia exhibit differential susceptibility to CD8 T cell mediated effector mechanisms during neurotropic coronavirus infection. In contrast to microglia, oligodendroglia are resistant to CD8 T cell perforin‐mediated viral control in the absence of IFNγ. Kinetic induction of MHC Class I expression by microglia and oligodendroglia in vivo was thus analyzed to assess responses to distinct inflammatory signals. Flow cytometry demonstrated delayed Class I surface expression by oligodendroglia compared with microglia. Distinct kinetics of Class I protein upregulation correlated with cell type specific transcription patterns of genes encoding Class I heavy chains and antigen processing components. Microglia isolated from naïve mice expressed high levels of these mRNAs, whereas they were near detection limits in oligodendroglia; nevertheless, Class I protein was undetectable on both cell types. Infection induced modest mRNA increases in microglia, but dramatic transcriptional upregulation in oligodendroglia coincident with IFNα or IFNγ mRNA increases in infected tissue. Ultimately mRNAs reached similar levels in both cell types at their respective time points of maximal Class I expression. Expression of Class I on microglia, but not oligodendroglia, in infected IFNγ deficient mice supported distinct IFN requirements for Class I presentation. These data suggest an innate immune preparedness of microglia to present antigen and engage CD8 T cells early following infection. The delayed, yet robust, IFNγ dependent capacity of oligodendroglia to express Class I suggests strict control of immune interactions to avoid CD8 T cell recognition and potential presentation of autoantigen to preserve myelin maintenance. © 2008 Wiley‐Liss, Inc.


title: Alterations in Nerve-Evoked Bladder Contractions in a Coronavirus-Induced Mouse Model of Multiple Sclerosis

text: BACKGROUND: Patients with neurodegenerative diseases such as multiple sclerosis, Parkinson’s, and Alzheimer’s often present with lower urinary tract symptoms (LUTS, urinary frequency, urgency, nocturia and retention) resulting from damage to the peripheral and central nervous systems. These studies were designed to examine the changes in the function of the bladder that may underlie neurogenic bladder dysfunction using a mouse model of demyelination in the CNS. METHODS: Bladders from 12 week old male C57BL/6J mice with coronavirus-induced encephalomyelitis (CIE, a chronic, progressive demyelinating disease model of human MS), and age-matched controls, were cut into 5–7 strips and suspended in physiological muscle baths for tension measurement in response to agonists and electric field stimulation (EFS). Experiments were performed on intact and denuded (with mucosa removed) bladder strips. RESULTS: The maximum effect of EFS was not significantly different between CIE and control bladders. Nerve-evoked EFS contractions (tetrodotoxin-sensitive) were blocked by a combination of atropine (cholinergic antagonist) and α,β-methylene ATP (an ATP analog that desensitizes purinergic receptors). In response to EFS, the α,β-methylene ATP-resistant (cholinergic) component of contraction was significantly reduced, while the atropine-resistant (purinergic) component was significantly increased in CIE bladders. Removal of the mucosa in CIE bladders restored the cholinergic component. Bethanechol (muscarinic receptor agonist) potency was significantly increased in CIE bladders. CONCLUSIONS: Our data demonstrate a deficit in the nerve-evoked cholinergic component of contraction that is not due to the ability of the smooth muscle to respond to acetylcholine. We conclude that neurodegenerative bladder dysfunction in this model of multiple sclerosis may be due, in part, to pathologic changes in the mucosa that causes suppression of muscarinic receptor-mediated contractile response and augmentation of purinergic response of the underlying muscle. Further studies utilizing CIE mice should help elucidate the pathological changes in the mucosa resulting from demyelination in the CNS.


title: Evaluation of viral co-infections in hospitalized and non-hospitalized children with respiratory infections using microarrays

text: The impact of viral co-infections and recently discovered viruses on the epidemiology of respiratory infections in children is still unclear. To simultaneously detect viruses that are involved in the aetiology of respiratory infections, we used a DNA/RNA microarray assay that identifies 17 different viruses or viral subtypes. Rhinopharyngeal washes were taken from 611 children (aged 1 month to 14 years) who presented in the emergency department with respiratory infections from June 2010 to June 2011 and were treated as outpatients (299, 48.9%) or hospitalized (312, 51.1%). Lower respiratory tract infection was diagnosed more often in hospitalized children (68% versus 36%, p 0.001). Of 397 children in which microarrays detected viral infection (70.1%), a single virus was found in 228 (57.4%) and two or more viruses in 169 (42.5%). The most prevalent viruses among children with positive samples were respiratory syncytial virus (RSV) in 225 (56.6%), parainfluenza virus (PIV) in 118 (29.7%), rhinovirus (RV) in 73 (18.4%), followed by influenza in 56 (14.1%), adenoviruses in 31 (7.8%), bocavirus in 25 (6.3%), human metapneumovirus in 15 (3.7%) and enteroviruses in 12 (3%). Most common viral co-infections were RSVA–RSVB in 46 children (27.2%), RSV–Influenza in 20 (11.8%), RSV–RV in 18 (10.6%) and PIV–RV in 13 (7.7%). Multiple logistic regression analysis revealed that viral co-infections were associated with increased probability for hospitalization (OR 1.52, 95% CI 1.01–2.29, p 0.04), and previous pneumococcal vaccination was associated with decreased probability for hospitalization (OR 0.52, 95% CI 0.33–0.81, p 0.004). We conclude that viral co-infections are involved in a significant proportion of children with an acute respiratory infection and may increase the severity of clinical presentation and the risk for hospitalization.


title: Impact of the Coronavirus Disease 2019 (COVID-19) Pandemic on Imaging Case Volumes

text: OBJECTIVE: The coronavirus disease 2019 (COVID-19) pandemic has had significant economic impact on radiology with markedly decreased imaging case volumes. The purpose of this study was to quantify the imaging volumes during the COVID-19 pandemic across patient service locations and imaging modality types. METHODS: Imaging case volumes in a large health care system were retrospectively studied, analyzing weekly imaging volumes by patient service locations (emergency department, inpatient, outpatient) and modality types (x-ray, mammography, CT, MRI, ultrasound, interventional radiology, nuclear medicine) in years 2020 and 2019. The data set was split to compare pre-COVID-19 (weeks 1-9) and post-COVID-19 (weeks 10-16) periods. Independent-samples t tests compared the mean weekly volumes in 2020 and 2019. RESULTS: Total imaging volume in 2020 (weeks 1-16) declined by 12.29% (from 522,645 to 458,438) compared with 2019. Post-COVID-19 (weeks 10-16) revealed a greater decrease (28.10%) in imaging volumes across all patient service locations (range 13.60%-56.59%) and modality types (range 14.22%-58.42%). Total mean weekly volume in 2020 post-COVID-19 (24,383 [95% confidence interval 19,478-29,288]) was statistically reduced (P = .003) compared with 33,913 [95% confidence interval 33,429-34,396] in 2019, across all patient service locations and modality types. The greatest decline in 2020 was seen at week 16 specifically for outpatient imaging (88%) affecting all modality types: mammography (94%), nuclear medicine (85%), MRI (74%), ultrasound (64%), interventional (56%), CT (46%), and x-ray (22%). DISCUSSION: Since the duration of COVID-19 pandemic remains uncertain, these results may assist in guiding short-term and long-term practice decisions based on the magnitude of imaging volume decline across different patient service locations and specific imaging modality types.


title: Genome Sequences of Porcine Epidemic Diarrhea Virus: In Vivo and In Vitro Phenotypes

text: Since the outbreak of porcine epidemic diarrhea virus (PEDV) in May 2013, U.S. swine producers have lost almost five million baby pigs. In an attempt to understand the evolution of PEDV in the United States and possibly develop a control strategy, we compared the genome sequences of a PEDV strain isolated from an infected piglet against its in vitro adapted version. The original PEDV strain was grown in Vero cells and passed 10 times serially in a MARC145 cell line. The sequence analysis of the native PEDV strain and in vitro passaged virus shows that the cell culture adaptation specifically modifies PEDV spike protein whereas the open reading frame 1a/b (ORF1a/b)-encoded polyprotein, the nucleoprotein, NS3B (ORF3), and membrane and envelope proteins remain unchanged.


title: Correlation between Heart fatty acid binding protein and severe COVID-19: A case-control study

text: BACKGROUND: Heart-fatty acid binding protein (HFABP) has been recognized as a highly heart-specific marker. However, it is currently unknown that its HFABP is also closely related to the severity of COVID-19. METHODS: We retrospectively screened 46 patients who met our inclusion criteria within 4 weeks. They were tested for HFABP after the diagnosis of COVID-19, and monitored for HFABP during their hospital stay. We tracked the patients during their hospital stay to determine if they had severe COVID-19 or mild-to-severe transition features. We calculated the chi-square test values found for HFABP to predict the correlation between HFABP levels and the severity of the COVID-19. RESULTS: Of these 46 cases, 16 cases with confirmed COVID-19 were tested for HFABP> 7 ng / mL upon admission; among them, 14 cases were diagnosed with severe COVID-19 within the hospitalization. The Odds ratio of the measured HFABP elevation was 6.81(95% confidence interval [CI] 5.23–8.40), and 3 patients with severe COVID-19 progressed in 5 patients with mild HFABP> 7 ng/mL. CONCLUSION: These data indicate that the elevation of HFABP is closely related to the severity of COVID-19 in the patients, and the elevated HFABP may cause rapid development of patients with mild COVID-19 into severe COVID-19. But serum HFABP negative maybe make patients with mild COVID-19 safer, the current data show no effect on the all-cause mortality. TRIAL REGISTRATION: Our study has been registered with the Chinese Clinical Trial Registry, the registration number: ChiCTR2000029829.


title: Strategies to Promote Social Connections Among Older Adults During ‘Social Distancing’ Restrictions

text: Older age and medical comorbidity are factors associated with more severe illness and risk of death due to COVID-19 infection. Social distancing is an important public health strategy for controlling the spread of the virus and minimizing its impact on the older adult population. It comes at a cost, however. Loneliness is associated with myriad adverse health outcomes, one of which is impaired immune functioning, which adds even greater risk for corona virus infection, complications and death. Older adults, therefore, are at compound risk, making effective management of loneliness and social isolation in our older patients a high priority target for preventive intervention. In this paper, we describe a cognitive-behavioral framework for social connectedness, including evidence-informed strategies clinicians can use to help patients develop a ‘Connections Plan’ to stay connected and promote their social, mental, and physical health during ‘social distancing’ restrictions. This set of strategies can be provided during brief (30 minute) telephone sessions and is analogous to creating a ‘Safety Plan’ for suicide risk. The approach is illustrated with three case examples.


Placental abruption in a twin pregnancy at 32 weeks’ gestation complicated by COVID-19, without vertical transmission to the babies.


Phenomenology and outcomes of in‐patients with Parkinson's disease during COVID‐19 pandemic


title: Viral and cellular RNA helicases as antiviral targets

text: Although there has been considerable progress in the development of antiviral agents in recent years, there is still a pressing need for new drugs both to improve on the properties of existing agents and to combat the problem of viral resistance. Helicases, both viral and human, have recently emerged as novel targets for the treatment of viral infections. Here, we discuss the role of these enzymes, factors affecting their potential as drug targets and progress in the development of agents that inhibit their activity using the hepatitis C virus-encoded helicase NS3 and the cellular helicase DDX3 adopted for use by HIV-1 as examples.


title: Rapid Uptake of Home-Based HIV Self-testing During Social Distancing for SARS-CoV2 Infection in Oregon

text: We implemented a pilot home HIV self-testing program one week after a stay-home order for SARS-CoV2 was enacted in Oregon. We advertised the program on a geospatial networking app and community partner websites targeting men who have sex with men; nine percent of web visits resulted in an order. Over 70% of the kits initially allotted to the program were ordered in the first 24 h of launch. One-third of participants had never tested for HIV. We found enthusiasm for discreet, free, home-based testing and uncovered an unmet need for HIV testing as clinical and outreach programs shuttered in Oregon.


title: Identification of a novel linear B-cell epitope in the UL26 and UL26.5 proteins of Duck Enteritis Virus

text: BACKGROUND: The Unique Long 26 (UL26) and UL26.5 proteins of herpes simplex virus are known to function during the assembly of the viruses. However, for duck enteritis virus (DEV), which is an unassigned member of the family Herpesviridae, little information is available about the function of the two proteins. In this study, the C-terminus of DEV UL26 protein (designated UL26c), which contains the whole of UL26.5, was expressed, and the recombinant UL26c protein was used to immunize BALB/c mice to generate monoclonal antibodies (mAb). The mAb 1C8 was generated against DEV UL26 and UL26.5 proteins and used subsequently to map the epitope in this region. Both the mAb and its defined epitope will provide potential tools for further study of DEV. RESULTS: A mAb (designated 1C8) was generated against the DEV UL26c protein, and a series of 17 partially overlapping fragments that spanned the DEV UL26c were expressed with GST tags. These peptides were subjected to enzyme-linked immunosorbent assay (ELISA) and western blotting analysis using mAb 1C8 to identify the epitope. A linear motif, (520)IYYPGE(525), which was located at the C-terminus of the DEV UL26 and UL26.5 proteins, was identified by mAb 1C8. The result of the ELISA showed that this epitope could be recognized by DEV-positive serum from mice. The (520)IYYPGE(525 )motif was the minimal requirement for reactivity, as demonstrated by analysis of the reactivity of 1C8 with several truncated peptides derived from the motif. Alignment and comparison of the 1C8-defined epitope sequence with those of other alphaherpesviruses indicated that the motif (521)YYPGE(525 )in the epitope sequence was conserved among the alphaherpesviruses. CONCLUSION: A mAb, 1C8, was generated against DEV UL26c and the epitope-defined minimal sequence obtained using mAb 1C8 was (520)IYYPGE(525). The mAb and the identified epitope may be useful for further study of the design of diagnostic reagents for DEV.


title: The Role of Severe Acute Respiratory Syndrome (SARS)-Coronavirus Accessory Proteins in Virus Pathogenesis

text: A respiratory disease caused by a novel coronavirus, termed the severe acute respiratory syndrome coronavirus (SARS-CoV), was first reported in China in late 2002. The subsequent efficient human-to-human transmission of this virus eventually affected more than 30 countries worldwide, resulting in a mortality rate of ~10% of infected individuals. The spread of the virus was ultimately controlled by isolation of infected individuals and there has been no infections reported since April 2004. However, the natural reservoir of the virus was never identified and it is not known if this virus will re-emerge and, therefore, research on this virus continues. The SARS-CoV genome is about 30 kb in length and is predicted to contain 14 functional open reading frames (ORFs). The genome encodes for proteins that are homologous to known coronavirus proteins, such as the replicase proteins (ORFs 1a and 1b) and the four major structural proteins: nucleocapsid (N), spike (S), membrane (M) and envelope (E). SARS-CoV also encodes for eight unique proteins, called accessory proteins, with no known homologues. This review will summarize the current knowledge on SARS-CoV accessory proteins and will include: (i) expression and processing; (ii) the effects on cellular processes; and (iii) functional studies.


title: Mutterschutz – auch im Zeitalter der Coronakrise

text: Pregnant employees should be protected, particularly in crisis situations. The Maternity Protection Act states that employees are not allowed to have contact with infectious people, including people with SARS-CoV-2 infections; no new regulation is required here.


title: Identification of critical determinants on ACE2 for SARS-CoV entry and development of a potent entry inhibitor

text: Abstract Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus, SARS-CoV. Virus entry into cells is mediated through interactions between spike (S) glycoprotein and angiotensin-converting enzyme 2 (ACE2). Alanine scanning mutagenesis analysis was performed to identify determinants on ACE2 critical for SARS-CoV infection. Results indicated that charged amino acids between residues 22 and 57 were important, K26 and D30, in particular. Peptides representing various regions of ACE2 critical for virus infection were chemically synthesized and evaluated for antiviral activity. Two peptides (a.a. 22–44 and 22–57) exhibited a modest antiviral activity with IC50 of about 50 μM and 6 μM, respectively. One peptide comprised of two discontinuous segments of ACE2 (a.a. 22–44 and 351–357) artificially linked together by glycine, exhibited a potent antiviral activity with IC50 of about 0.1 μM. This novel peptide is a promising candidate as a therapeutic agent against this deadly emerging pathogen.


Border screening for SARS


title: Infectious bronchitis virus: immunopathogenesis of infection in the chicken

text: The immunopathogenesis of infectious bronchitis virus (IBV) infection in the chicken is reviewed. While infectious bronchitis (IB) is considered primarily a disease of the respiratory system, different IBV strains may show variable tissue tropisms and also affect the oviduct and the kidneys, with serious consequences. Some strains replicate in the intestine but apparently without pathological changes. Pectoral myopathy has been associated with an important recent variant. Several factors can influence the course of infection with IBV, including the age, breed and nutrition of the chicken, the environment and intercurrent infection with other infectious agents. Immunogenic components of the virus include the S (spike) proteins and the N nucleoprotein. The humoral, local and cellular responses of the chicken to IBV are reviewed, together with genetic resistance of the chicken. In long-term persistence of IBV, the caecal tonsil or kidney have been proposed as the sites of persistence. Antigenic variation among IBV strains is related to relatively small differences in amino acid sequences in the S1 spike protein. However, antigenic studies alone do not adequately define immunological relationships between strains and cross-immunisation studies have been used to classify IBV isolates into ‘protectotypes’. It has been speculated that changes in the S1 protein may be related to differences in tissue tropisms shown by different strains. Perhaps in the future, new strains of IBV may arise which affect organs or systems not normally associated with IB.


title: Approaching COVID‐19 ‐ bedside strategies for intensive care

text: The spread of novel coronavirus disease (COVID‐19) urged a never‐seen coordinated global response to prepare the health system, including primary care, hospital facilities and intensive care units (ICUs). Lessons have been learned from countries who suffered the pandemic at the beginning, helping the ones which are on different phases of the spreading curve. Currently, optimizing intensive care resources is mandatory as admittance to the ICUs remains rising exponentially. While public and private health system struggle for changing the slope of the curve, intensivists prepare the facilities for a tsunami of respiratory failure patients with COVID‐19


title: Acute‐onset smell and taste disorders in the context of COVID‐19: a pilot multicentre polymerase chain reaction based case–control study

text: BACKGROUND AND PURPOSE: Specific respiratory tract infections, including COVID‐19, may cause smell and/or taste disorders (STDs) with increased frequency. The aim was to determine whether new‐onset STDs are more frequent amongst COVID‐19 patients than influenza patients. METHOD: This was a case–control study including hospitalized patients of two tertiary care centres. Consecutive patients positive for COVID‐19 polymerase chain reaction (cases) and patients positive for influenza polymerase chain reaction (historical control sample) were assessed during specific periods, employing a self‐reported STD questionnaire. RESULTS: Seventy‐nine cases and 40 controls were included. No significant differences were found in basal features between the two groups. New‐onset STDs were significantly more frequent amongst cases (31, 39.2%) than in the control group (5, 12.5 %) [adjusted odds ratio 21.4 (2.77–165.4, P = 0.003)]. COVID‐19 patients with new‐onset STDs were significantly younger than COVID‐19 patients without STDs (52.6 ± 17.2 vs. 67.4 ± 15.1, P < 0.001). Amongst COVID‐19 patients who presented STDs, 22 (70.9%) recalled an acute onset and it was an initial manifestation in 11 (35.5%). Twenty‐five (80.6%) presented smell disorders (mostly anosmia, 14, 45.2%) and 28 (90.3%) taste disorders (mostly ageusia, 14, 45.2%). Only four (12.9 %) reported concomitant nasal obstruction. The mean duration of STD was 7.5 ± 3.2 days and 12 patients (40%) manifested complete recovery after 7.4 ± 2.3 days of onset. CONCLUSION: New‐onset STDs were significantly more frequent amongst COVID‐19 patients than influenza patients; they usually had an acute onset and were commonly an initial manifestation. The use of STD assessment in anamnesis as a hint for COVID‐19 and to support individuals’ self‐isolation in the current epidemic context is suggested.


“Re: CoViD-19 and ortho and trauma surgery: The Italian experience” (In Press)


title: China's post-1978 experience in outbound tourism

text: Abstract In 1978, Deng Xiao-Ping introduced economic reforms and the open-door policy in China. Since then, the Chinese government started to recognize tourism as a means to economic development and modernisation. Chinese outbound tourism has been managed and regulated by the Approved Destination Status (ADS) system, which is based on bilateral tourism agreement between China and overseas destinations. The ratio-to-moving average technique and Box–Jenkins (1970) univariate time series modelling are used to analyse Chinese tourist arrival patterns to Australia for the period 1984–2004. Tests for stationarity in the time series of tourist arrivals are also conducted.


title: Mining the Characteristics of COVID-19 Patients in China: Analysis of Social Media Posts

text: BACKGROUND: In December 2019, pneumonia cases of unknown origin were reported in Wuhan City, Hubei Province, China. Identified as the coronavirus disease (COVID-19), the number of cases grew rapidly by human-to-human transmission in Wuhan. Social media, especially Sina Weibo (a major Chinese microblogging social media site), has become an important platform for the public to obtain information and seek help. OBJECTIVE: This study aims to analyze the characteristics of suspected or laboratory-confirmed COVID-19 patients who asked for help on Sina Weibo. METHODS: We conducted data mining on Sina Weibo and extracted the data of 485 patients who presented with clinical symptoms and imaging descriptions of suspected or laboratory-confirmed cases of COVID-19. In total, 9878 posts seeking help on Sina Weibo from February 3 to 20, 2020 were analyzed. We used a descriptive research methodology to describe the distribution and other epidemiological characteristics of patients with suspected or laboratory-confirmed SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) infection. The distance between patients’ home and the nearest designated hospital was calculated using the geographic information system ArcGIS. RESULTS: All patients included in this study who sought help on Sina Weibo lived in Wuhan, with a median age of 63.0 years (IQR 55.0-71.0). Fever (408/485, 84.12%) was the most common symptom. Ground-glass opacity (237/314, 75.48%) was the most common pattern on chest computed tomography; 39.67% (167/421) of families had suspected and/or laboratory-confirmed family members; 36.58% (154/421) of families had 1 or 2 suspected and/or laboratory-confirmed members; and 70.52% (232/329) of patients needed to rely on their relatives for help. The median time from illness onset to real-time reverse transcription-polymerase chain reaction (RT-PCR) testing was 8 days (IQR 5.0-10.0), and the median time from illness onset to online help was 10 days (IQR 6.0-12.0). Of 481 patients, 32.22% (n=155) lived more than 3 kilometers away from the nearest designated hospital. CONCLUSIONS: Our findings show that patients seeking help on Sina Weibo lived in Wuhan and most were elderly. Most patients had fever symptoms, and ground-glass opacities were noted in chest computed tomography. The onset of the disease was characterized by family clustering and most families lived far from the designated hospital. Therefore, we recommend the following: (1) the most stringent centralized medical observation measures should be taken to avoid transmission in family clusters; and (2) social media can help these patients get early attention during Wuhan’s lockdown. These findings can help the government and the health department identify high-risk patients and accelerate emergency responses following public demands for help.


title: Flaviviral methyltransferase/RNA interaction: Structural basis for enzyme inhibition

text: Abstract Flaviviruses are the causative agents of severe diseases such as Dengue or Yellow fever. The replicative machinery used by the virus is based on few enzymes including a methyltransferase, located in the N-terminal domain of the NS5 protein. Flaviviral methyltransferases are involved in the last two steps of the mRNA capping process, transferring a methyl group from S-adenosyl-l-methionine onto the N7 position of the cap guanine (guanine-N7 methyltransferase) and the ribose 2′O position of the first nucleotide following the cap guanine (nucleoside-2′O methyltransferase). The RNA capping process is crucial for mRNA stability, protein synthesis and virus replication. Such an essential function makes methyltransferases attractive targets for the design of antiviral drugs. In this context, starting from the crystal structure of Wesselsbron flavivirus methyltransferase, we elaborated a mechanistic model describing protein/RNA interaction during N7 methyl transfer. Next we used an in silico docking procedure to identify commercially available compounds that would display high affinity for the methyltransferase active site. The best candidates selected were tested in vitro to assay their effective inhibition on 2′O and N7 methyltransferase activities on Wesselsbron and Dengue virus (Dv) methyltransferases. The results of such combined computational and experimental screening approach led to the identification of a high-potency inhibitor.


title: Evaluation of decontamination methods for commercial and alternative respirator and mask materials – view from filtration aspect

text: This study aims to evaluate the filtration performance of three commercially available (3M 8210 respirator, Halyard 48,207 surgical mask, and 3M 1820 procedure mask) and two alternative face mask and respirator materials (Halyard H600 sterilization wrap and Cummins EX101) after selected decontamination treatments, including isopropanol (IPA) treatments (soaking or spraying), ultraviolet germicidal irradiation (UVGI), and heat treatments (dry heat at 77 °C or steam heat). Both IPA soaking and spraying removed most electrostatic charges on all four electret materials (three commercial and one alternative), causing significant deterioration of filtration efficiency to unacceptable level. The other non-electret alternative material sustained its N95-grade performance after both IPA soaking and spraying treatments, demonstrating the possible application of IPA disinfection for non-electret alternative respirator/mask materials. UVGI preserved the filtration of all three commercially available respirator/mask materials after up to 10 treatments, suggesting it can be a possible decontamination method for hospital and clinic use without compromising respirator/mask performance. The considerations of the practical implementation of this method was discussed. Between the two heat treatment methods tested, dry heat showed better compatibility with electret material by sustaining both filtration efficiency and fit (tested on commercial respirator only), although adding moisture was reported in favor of virus inactivation. Heat treatment is easily accessible method for general publics to implement at home, while it is recommended to maintain the moisture level below saturation. Comparing to size-integrated method, the size-resolved fractional efficiency measurement technique, although more time consuming, proved to be a better method for evaluating respirator/mask filtration performance after decontaminations by providing more sensitive detection of performance degradation and the capability of distinguishing charge loss to other mechanisms causing efficiency deterioration. Detailed descriptions are provided in methodology part to emphasize the cares needed for an appropriate efficiency evaluation. The limited results in this study on worn masks made of alternative sterilization wrap indicated possible performance degradation of electret material caused by normal human wearing activities, suggesting the need of assessing respirator/mask decontamination strategy by testing practically worn-and-decontaminated/reused samples instead of unworn only-decontaminated counterparts.


Organización de un Servicio de Dermatología en la pandemia del COVID-19


Sars Coronavirus Accessory ORFs Encode Luxury Functions


title: The mystery of “missing” visits in an emergency cardiology department, in the era of COVID-19.; a time-series analysis in a tertiary Greek General Hospital

text: BACKGROUND: In the era of the current COVID-19 health crisis, the aim of the present study was to explore population behavior as regards the visits in the Εmergency Cardiology department (ECD) of a tertiary General Hospital that does not hospitalize SARS-CoV-2 infected patients METHODS AND RESULTS: Daily number of visits at the EDC and admissions to Cardiology Wards and Intensive Care Unit of a tertiary General Hospital, in Athens, Greece, were retrieved from hospital’s database (January 1st–April 30th 2018, 2019 and 2020). A highly significant reduction in the visits at ECD of the hospital during March and April 2020 was observed as compared with January and February of the same year (p for linear trend < ·001); in particular the number of visits was 41.1% lower in March 2020 and 32.7% lower in April 2020, as compared to January 2020. As the number of confirmed COVID-19 cases throughout the country increased (i.e., from February 26th to April 2nd) the number of visits at ECD decreased (p = 0.01), whereas, the opposite was observed in the period afterwards (p = 0.01).The number of acute Myocardial infarctions (MI) cases in March 2020 was the lowest compared to the entire three year period (p < 0·001); however, the number of acute MI cases in April 2020 was doubled as compared to March 2020, but still was lower than the preceding years (p < 0·001). CONCLUSIONS: It is hard to explain the mystery of the “missing” emergency hospital visits. However, if this decline in cardiovascular disease related hospital visits is “true”, it is something that needs to be rigorously studied, to learn how to keep these rates down.


title: Prognostication and Proactive Planning in COVID-19

text: Abstract Accurate prognostication is challenging in the setting of SARS-CoV-2, the virus responsible for COVID-19, due to rapidly changing data, studies that are not generalizable and lack of morbidity and functional outcomes in survivors. To provide meaningful guidance to patients, existing mortality data must be considered and appropriately applied. While most people infected with SARS-CoV-2 will recover, mortality increases with age and co-morbidity in those who develop severe illness.


title: Transmembrane Domains of Highly Pathogenic Viral Fusion Proteins Exhibit Trimeric Association In Vitro

text: Enveloped viruses require viral fusion proteins to promote fusion of the viral envelope with a target cell membrane. To drive fusion, these proteins undergo large conformational changes that must occur at the right place and at the right time. Understanding the elements which control the stability of the prefusion state and the initiation of conformational changes is key to understanding the function of these important proteins. The construction of mutations in the fusion protein transmembrane domains (TMDs) or the replacement of these domains with lipid anchors has implicated the TMD in the fusion process. However, the structural and molecular details of the role of the TMD in these fusion events remain unclear. Previously, we demonstrated that isolated paramyxovirus fusion protein TMDs associate in a monomer-trimer equilibrium, using sedimentation equilibrium analytical ultracentrifugation. Using a similar approach, the work presented here indicates that trimeric interactions also occur between the fusion protein TMDs of Ebola virus, influenza virus, severe acute respiratory syndrome coronavirus (SARS CoV), and rabies virus. Our results suggest that TM-TM interactions are important in the fusion protein function of diverse viral families. IMPORTANCE Many important human pathogens are enveloped viruses that utilize membrane-bound glycoproteins to mediate viral entry. Factors that contribute to the stability of these glycoproteins have been identified in the ectodomain of several viral fusion proteins, including residues within the soluble ectodomain. Although it is often thought to simply act as an anchor, the transmembrane domain of viral fusion proteins has been implicated in protein stability and function as well. Here, using a biophysical approach, we demonstrated that the fusion protein transmembrane domains of several deadly pathogens—Ebola virus, influenza virus, SARS CoV, and rabies virus—self-associate. This observation across various viral families suggests that transmembrane domain interactions may be broadly relevant and serve as a new target for therapeutic development.


title: Lopinavir/ritonavir and interferon combination therapy may help shorten the duration of viral shedding in patients with COVID‐19: a retrospective study in two designated hospitals in Anhui, China

text: BACKGROUND: Prolonged viral shedding may pose a threat to the control of coronavirus disease 2019 (COVID‐19), and data on the duration of SARS‐CoV‐2 shedding are still limited, with the associated factors being unknown. METHODS: All adult patients with laboratory‐confirmed COVID‐19 were included in this retrospective cross‐sectional study in two designated hospitals during 21 January 2020‐16 March 2020 in Anhui, China. In all patients, data on the duration of SARS‐CoV‐2 RNA shedding were analyzed by reviewing all RNA detection results during hospitalization. Additionally, demographic, clinical, treatment, laboratory and outcome data were also collected from electronic medical records. Factors associated with prolonged viral shedding were analyzed with the Cox proportional hazards model. RESULTS: Among 181 patients, the mean age was 44.3±13.2 years, and 55.2% were male. The median duration of viral shedding from illness onset was 18.0 days (IQR 15.0‐24.0). Prolonged viral shedding was associated with longer hospital stays (P<0.001) and higher medical costs (P<0.001). The severity of COVID‐19 had nothing to do with prolonged shedding. Moreover, the median time from onset to antiviral treatment initiation was 5.0 days (IQR 3.0‐7.0). Delayed antiviral treatment (HR 0.976 [95% CI, 0.962‐0.990]) and lopinavir/ritonavir + IFN‐α combination therapy as the initial antiviral treatment (HR 1.649 [95% CI, 1.162‐2.339]) were independent factors associated with prolonged SARS‐CoV‐2 RNA shedding. CONCLUSIONS: SARS‐CoV‐2 showed prolonged viral shedding, causing increased hospital stays and medical costs. Early initiation of lopinavir/ritonavir + IFN‐α combination therapy may help shorten the duration of SARS‐CoV‐2 shedding. This article is protected by copyright. All rights reserved.


title: The power of detention in the management of non-compliance with tuberculosis treatment: A survey of Irish practitioners and analysis of potential legal liability

text: SUMMARY Objectives The detention of patients infected with tuberculosis has recently been the subject of significant professional and public interest. In Ireland, the power to detain and isolate probable sources of infectious disease is found in the Health Act 1947. The objective of this study was to describe the use of the power to detain, and to examine relevant legal implications. Study design Cohort survey. Methods Respiratory and infectious disease physicians practising in the public sector were invited to complete a self-administered postal questionnaire. Results Of the 44 clinicians surveyed, 33 responded, representing a total of 356 years of specialist practice (mean 10.8 years). Although 70% of respondents had made use of threats of formal detention in dealing with non-compliant patients, only one formal detention under statutory powers was identified. Infrastructural and legal concerns with the use of detention were common. There was widespread support for a broadening of the range of additional public health powers, including a power of prolonged detention in the setting of multi-drug-resistant tuberculosis. Conclusion Detention and isolation of non-compliant tuberculosis patients remains in active use. Physicians detaining, or threatening to detain, patients continue to expose themselves to legal liability because of the outdated legal framework underlying those powers.


title: Viruses and the cellular RNA decay machinery

text: The ability to control cellular and viral gene expression, either globally or selectively, is central to a successful viral infection, and it is also crucial for the host to respond and eradicate pathogens. In eukaryotes, regulation of message stability contributes significantly to the control of gene expression and plays a prominent role in the normal physiology of a cell as well as in its response to environmental and pathogenic stresses. Not surprisingly, emerging evidence indicates that there are significant interactions between the eukaryotic RNA turnover machinery and a wide variety of viruses. Interestingly, in many cases viruses have evolved mechanisms not only to evade eradication by these pathways, but also to manipulate them for enhanced viral replication and gene expression. Given our incomplete understanding of how many of these pathways are normally regulated, viruses should be powerful tools to help deconstruct the complex networks and events governing eukaryotic RNA stability. Copyright © 2010 John Wiley & Sons, Ltd. 1.. RNA Turnover and Surveillance > Turnover/Surveillance Mechanisms; 2.. RNA in Disease and Development > RNA in Disease.


A first case of Mild Encephalitis with Reversible splenial lesions (MERS) as a presenting feature of SARS-CoV-2


title: A 28‐year study of human parainfluenza in Rio Grande do Sul, Southern Brazil

text: PROBLEM: Human parainfluenza virus (hPIV) is an important pathogen in respiratory infections, however the health burden of hPIV is underestimated. This study describes the infections by hPIV1‐3 in Rio Grande do Sul, Brazil, from 1990 to 2017, providing data of the frequency and seasonality of cases and associated clinical symptoms. METHOD OF STUDY: Nasopharyngeal samples of patients with respiratory infection were collected, clinical data were analyzed, and immunofluorescence was used to detect hPIV. RESULTS: Respiratory viruses were detected in 33.63% of respiratory infections. In a total of 11 606 cases of viral respiratory infection, 781 were positive for hPIV; hPIV prevalence ranged from 2.14% to 27% of viral respiratory infections. hPIV1 circulates mainly during fall; hPIV3 circulation, in turn, starts in fall and peaks during spring; and cases of hPIV2 are reported along the year, with peaks in fall and early spring. The most affected age group was children, with hPIV prevalence of 74.23% in patients for less than 1 year. A higher proportion of girls were infected than boys, however, no difference by sex was observed considering all age groups. The most frequent type was hPIV3, especially in hospitalized patients. Both hPIV1 and 3 were associated with dyspnea, while hPIV2 caused mild symptoms mainly in nonhospitalized patients. Nineteen fatalities occurred, 89.5% of them associated with risk factors (prematurity; chronic diseases; age, <1 or >60 years). CONCLUSION: hPIV causes a high number of respiratory infections, leading to hospitalization especially in children; epidemiological and surveillance studies are important for the control and management of respiratory infections.


title: The impact of the COVID-19 epidemic on the utilization of emergency dental services

text: Abstract Background/Purpose To assess how the current COVID-19 epidemic influenced peoples' utilization of emergency dental services in Beijing, China. Methods The first-visit patients seeking emergency dental services before or at the beginning of the COVID-19 epidemic were retrieved. Their demographic characteristics and the reasons for visiting were recorded and analyzed. Results There were 2,537 patients involved in this study. Thirty-eight percent fewer patients visited the dental urgency at the beginning of the COVID-19 epidemic than before. The distribution of dental problems has changed significantly. The proportion of dental and oral infection raised from 51.0% of pre-COVID-19 to 71.9% during COVID-19, and dental trauma decreased from 14.2% to 10.5%. Meanwhile, the non-urgency cases reduced to three-tenths of pre-COVID-19. Conclusion Within the limitation of this study, the COVID-19 epidemic had a strong influence on the utilization of emergency dental services.


title: Novel replication complex architecture in rubella replicon‐transfected cells

text: Rubella virus (RUB) assembles its replication complexes (RCs) in modified organelles of endo‐lysosomal origin, known as cytopathic vacuoles (CPVs). These peculiar structures are key elements of RUB factories, where rough endoplasmic reticulum, mitochondria, and Golgi are recruited. Bicistronic RUB replicons expressing an antibiotic resistance gene either in the presence or the absence of the RUB capsid (C) gene were used to study the structure of RCs in transfected cells. Confocal microscopy showed that the RUB replicase components P90 and P150 localized to CPVs, as did double‐stranded RNA (dsRNA), a marker for RNA synthesis. Electron microscopy (EM) showed that replicons generated CPVs containing small vesicles and large vacuoles, similar to CPVs from RUB‐infected cells and that the replicase proteins were sufficient for organelle recruitment. Some of these CPVs contained straight membranes. When cross‐sectioned, these rigid membranes appeared to be sheets of closely packed proteins. Immuno‐EM revealed that these sheets, apparently in contact with the cytosol, contained both P150 and P90, as well as dsRNA, and thus could be two‐dimensional arrays of functional viral replicases. Labelling of dsRNA after streptolysin‐O permeabilization showed that replication of viral genome takes place on the cytoplasmic side of CPVs. When present, C accumulated around CPVs. Mitochondrial protein P32 was detected within modified CPVs, the first demonstration of involvement of this protein, which interacts with C, with RCs.


title: Multiplex real-time PCR for detection of respiratory tract infections

text: Abstract Background Broad diagnostics of respiratory infection by molecular assays has not yet won acceptance due to technical difficulties and high costs. Objectives To evaluate clinical applicability of multiplex real-time PCR. Study design An assay targeting influenza virus A (IfA) and B (IfB), parainfluenza 1-3 (PIV), human metapneumovirus (MPV), respiratory syncytial virus (RSV), rhinovirus (RV), enterovirus (EV), adenovirus (AdV), human coronaviruses (229E, OC43, NL63), M. pneumoniae and Ch. pneumoniae was developed and run daily on consecutive clinical nasopharyngeal swab samples. Results An etiology was identified in 48% of the 954 samples, with IfA in 25%, RV in 20%, MPV in 10% and M. pneumoniae in 10% of the positive. By a rational procedure costs could be reduced and the customer price set relatively low (€33 per sample). Conclusion Streamlined testing and cost limitation is achievable and probably critical for implementation of a broad molecular diagnostics of respiratory infections.


title: Manejo de pacientes de Ortopedia y Traumatología en el contexto de la contingencia por covid-19: revisión de conceptos actuales

text: Resumen El objetivo del presente estudio es presentar una revisión de la literatura disponible, que permita abordar de forma ordenada la evidencia actual con respecto a la organización de un servicio de cirugía ortopédica y traumatología durante la pandemia por covid-19, el manejo de los pacientes y las medidas de protección para el personal de salud involucrado. Se realizó una búsqueda de literatura en bases de datos (PUBMED y Scielo), instituciones gubernamentales de salud y sociedades científicas. Los autores seleccionaron los artículos que consideraron pertinentes para la presente revisión. Se plantean recomendaciones para los distintos escenarios de la práctica ortopédica: consulta externa, hospitalización, cirugía (electiva y de urgencias) y urgencias; así como recomendaciones específicas para algunas subespecialidades (cirugía de columna, cirugía de mano y ortopedia pediátrica). Abstract In this study, we review current evidence on how to organize an orthopaedic’s department during the covid-19 pandemic, how to manage patients and how to protect health personnel. We executed these search in PUBMED, Scielo, government health agencies and scientific associations databases. We selected articles based on relevance. We made recommendations for different scenarios of orthopedic practice: outpatient clinic, inpatient care, surgery (elective and emergency) and emergency department. We also made recommendatios for some orthopaedic subspecialties, including spinal surgery, hand surgery and pediatric orthopaedics.


title: Mass spectrometry for monitoring protease reactions

text: More than 560 genes are annotated as proteases in the human genome. About half of the genes are not or are only marginally characterized. Over the past decade, mass spectrometry has become the basis for proteomics, especially for protein identification, performed in a high-throughput manner. This development was also very fruitful for exploring the complex systems associated with protease functions, as briefly reviewed here. Mass spectrometry is an ideal tool for monitoring protease reactions, as will be highlighted in this review.


title: Vaccination of mice with recombinant baculovirus expressing spike or nucleocapsid protein of SARS-like coronavirus generates humoral and cellular immune responses

text: Abstract Continuous efforts have been made to develop a prophylactic vaccine against severe acute respiratory syndrome coronavirus (SARS-CoV). In this study, two recombinant baculoviruses, vAc-N and vAc-S, were constructed, which contained the mammalian-cell activate promoter element, human elongation factor 1α-subunit (EF-1α), the human cytomegalovirus (CMV) immediate-early promoter, and the nucleocapsid (N) or spike (S) gene of bat SARS-like CoV (SL-CoV) under the control of the CMV promoter. Mice were subcutaneously and intraperitoneally injected with recombinant baculovirus, and both humoral and cellular immune responses were induced in the vaccinated groups. The secretion level of IFN-γ was much higher than that of IL-4 in vAc-N or vAc-S immunized groups, suggesting a strong Th1 bias towards cellular immune responses. Additionally, a marked increase of CD4 T cell immune responses and high levels of anti-SARS-CoV humoral responses were also detected in the vAc-N or vAc-S immunized groups. In contrast, there were significantly weaker cellular immune responses, as well as less antibody production than in the control groups. Our data demonstrates that the recombinant baculovirus can serve as an effective vaccine strategy. In addition, because effective SARS vaccines should act to not only prevent the reemergence of SARS-CoV, but also to provide cross-protection against SL-CoV, findings in this study may have implications for developing such cross-protective vaccines.


title: Chapter 6 Public Health, Communicable Diseases and Global Health

text: Public health professionals are considering how molecular medicine applications can impact on preventive medicine and health provision enabling interventions to start earlier. There is the potential for molecular medicine to improve the workplace although inappropriate use will lead to discrimination. The public health issue of communicable diseases is being addressed through molecular medicine in more effective and early detection and a better understanding of pathogenesis. A threat from emerging and re-emerging infections requires multiple approaches but early and definitive detection will need nucleic acid based approaches. The non-communicable diseases are now a global world problem, particularly obesity. While molecular medicine is still to make a significant impact on this disorder some interesting findings are emerging about the role of genes in obesity. Ultimately, global health issues must include more efficient and environmentally sustainable strategies. Molecular technologies provide a promising approach for the way forward. Keywords Epidemiology, workplace, diagnostics, resistance, virulence, emerging and re-emerging infections, obesity, bioeconomy


title: Discharged COVID‐19 Patients Testing Positive Again for SARS‐CoV‐2 RNA: A Minireview of Published Studies from China

text: In the ongoing COVID‐19 pandemic, one potential cause of concern is that some discharged COVID‐19 patients are testing positive again for SARS‐CoV‐2 RNA. To better understand what is happening and to provide public health policy planners and clinicians timely information, we have searched and reviewed published studies about discharged patients testing positive again for the SARS‐CoV‐2 RNA. Our search found 12 reports, all of which described patients in China. Our review of these reports indicates the presence of discharged patients who remain asymptomatic but test positive. However, it is unclear whether they are contagious because a positive RT‐PCR test does not necessarily indicate the presence of replicating and transmissible virus. Our review suggests the need for timely, parallel testing of different samples, including for example, fecal specimens, from COVID‐19 patients before and after they are discharged from hospitals. This article is protected by copyright. All rights reserved.


title: A Case Report of Suicide Attempt Caused by Acute and Transient Psychotic Disorder during the COVID-19 Outbreak

text: We reported a case of suicide attempt caused by acute and transient psychotic disorder during the COVID-19 outbreak, which broke out in December 2019 in Wuhan. An epidemic of infectious diseases brought great psychological pressure to the public. During this period, a 20-year-old man went to the hospital repeatedly because he suspected that he was infected, with suspicious auditory hallucinations, self-laughter, primary delusions, victimization delusions, relationship delusions, and suicide attempts. He was diagnosed with Acute Transient Psychotic Disorder. 0.1 g bid Quetiapine was given orally, then gradually increased to 0.4 g per day, supplemented by cognitive therapy. The patient was discharged from hospital in relief of symptoms on February 9th. Conclusion. During the epidemic period, in addition to strengthening the protection work, we should also monitor the mental and psychological state of the population to prevent mental illness caused by coronavirus.


title: Internal versus external determinants of Schistosoma japonicum transmission in irrigated agricultural villages

text: Currently schistosomiasis transmission has been suppressed to low levels in many historically endemic areas of China by widespread use of praziquantel in human and bovine populations and application of niclosamide for snail control. However, re-emergent transmission has signalled the need for sustainable interventions beyond these repeated chemical interventions. To take advantage of ongoing investment in rural infrastructure, an index of schistosomiasis transmission potential is needed to identify villages where environmental modifications would be particularly effective. Based on a retrospective analysis of data from 10 villages in Sichuan Province, an index linked to the basic reproductive number is shown to have promise in meeting this need. However, a lack of methods for estimating the spatial components of the proposed metric and for estimating the import of cercariae and miracidia from neighbouring villages leads to significant uncertainty in its estimation. These findings suggest a priority effort to develop methods for measuring the free-swimming forms of the parasite in surface waters. This need is underscored by the high cost and limited sensitivity of current methods for diagnosing human infection and mounting evidence of the inadequacy of snail surveys to identify environments supporting low levels of transmission.


title: Lack of Transmission among Close Contacts of Patient with Case of Middle East Respiratory Syndrome Imported into the United States, 2014

text: In May 2014, a traveler from the Kingdom of Saudi Arabia was the first person identified with Middle East respiratory syndrome coronavirus (MERS-CoV) infection in the United States. To evaluate transmission risk, we determined the type, duration, and frequency of patient contact among health care personnel (HCP), household, and community contacts by using standard questionnaires and, for HCP, global positioning system (GPS) tracer tag logs. Respiratory and serum samples from all contacts were tested for MERS-CoV. Of 61 identified contacts, 56 were interviewed. HCP exposures occurred most frequently in the emergency department (69%) and among nurses (47%); some HCP had contact with respiratory secretions. Household and community contacts had brief contact (e.g., hugging). All laboratory test results were negative for MERS-CoV. This contact investigation found no secondary cases, despite case-patient contact by 61 persons, and provides useful information about MERS-CoV transmission risk. Compared with GPS tracer tag recordings, self-reported contact may not be as accurate.


title: Pharmacokinetics of Favipiravir in Critically Ill Patients with COVID‐19

text: Since December 2019, a novel coronavirus (SARS‐CoV‐2) infection has been rapidly spreading worldwide and causing the respiratory illness, coronavirus disease 2019 (COVID‐19). The anti‐retroviral drug favipiravir (FPV) has been experimentally used for COVID‐19 treatment since March 2020 in Japan. However, the pharmacokinetics of FPV in critically ill patients is unknown. We measured the serum concentration of FPV using high‐performance liquid chromatography in patients with severe COVID‐19 who were admitted to the intensive care unit and placed on mechanical ventilation. The patients were administered 1600 mg of FPV twice daily on Day 1, followed by 600 mg twice daily from Day 2 to Day 5 (or more if needed). Suspensions of FPV tablets were administered through a nasogastric tube. Seven patients were enrolled in this study. Forty‐nine blood samples were obtained from the eligible patients to evaluate FPV concentration. The FPV trough (after 8–12 h) concentrations of most samples were lower than the lower limit of quantification (1 µg/mL) and EC(50) (9.7 µg/mL) against SARS‐CoV‐2 previously tested in vitro. FPV trough concentration in critically ill patients was much lower than that of healthy subjects in a previous clinical trial, which is a cause for great concern. Further study is required to determine the optimal strategy for treatment of patients with severe COVID‐19.


Sharing Cigarette Smoking and COVID-19 Outbreak in a Party Group


title: Structural and antigenic analysis of the yellow head virus nucleocapsid protein p20

text: Abstract Yellow head virus (YHV) is an invertebrate nidovirus that is highly pathogenic for marine shrimp. Nucleotide sequence analysis indicated that the YHV ORF2 gene encodes a basic protein (pI =9.9) of 146 amino acids with a predicted molecular weight of 16,325.5Da. The deduced amino acid sequence indicated a predominance of basic (15.1%), acidic (9.6%) and hydrophilic polar (34.3%) residues and a high proportion proline and glycine residues (16.4%). The ORF2 gene was cloned and expressed in Escherichia coli as a M r =21kDa His6-protein that reacted with YHV nucleoprotein (p20) monoclonal antibody. Segments representing the four linear quadrants of the nucleoprotein were also expressed in E. coli as GST-fusion proteins. Immunoblot analysis using YHV polyclonal rabbit antiserum indicated the presence of linear epitopes in all except the V37–Q74 quadrant. Immunoblot analysis of the GST-fusion proteins and C-terminally truncated segments of the nucleoprotein allowed mapping of YHV monoclonal antibodies Y19, Y20 and YII4 to linear epitopes in the acidic domain between amino acids I116 and E137. The full-length nucleoprotein was expressed at high level in E. coli and was easily purified in quantity from the soluble cell fraction by Ni+-NTA affinity chromatography.


title: Inhibitory effect of papaverine on RNA and protein synthesis of vesicular stomatitis virus

text: Abstract Papaverine, an inhibitor of cAMP phosphodiesterase, reduced yields of infectious vesicular stomatitis virus in HEp-2 cells approximately 100-fold if added to cultures at a concentration of 30 μM before and after virus infection. The extent of papaverine-induced suppression of viral growth was dependent on drug dose and treatment regimen. Cells progressively recovered their viral permissive state after removal of drug. The cyclic nucleotide, cGMP, nullified the inhibitory effect of papaverine if added to cells during drug treatment. Pulse labeling experiments with [35S]methionine showed that papaverine compromises production of all virus-specific proteins in infected cells without adversely affecting host cell protein synthesis. Treatment of cells with papaverine strongly inhibited the production of viral RNA and both cellular RNA and DNA. It was found that VSV causes an immediate but transient stimulation of DNA synthesis in HEp-2 cells which is prevented by papaverine treatment. This drug also selectively blocked primary transcription of VSV in vivo and to a lesser extent in vitro RNA polymerase activity of the virion-bound transcriptase. The finding that papaverine has a strong inhibitory effect on viral biosynthesis including early transcription suggests that VSV replication may depend on host factors that regulate intracellular levels of cyclic nucleotides such as cAMP.


title: A glimpse into the future: can we learn lessons from the pandemic?

text: This month's installment of “The AJT Report” explores several ways in which COVID‐19 may be presenting opportunities for creating meaningful change for the transplant community and the US health system as a whole.


title: Biology

text: The hottest developments in the fields of cancer research, neuroscience, genomics and proteomics, anti-virals and more, with a pick of the key research papers in these areas.


title: Interaction of coronavirus nucleocapsid protein with the 5′‐ and 3′‐ends of the coronavirus genome is involved in genome circularization and negative‐strand RNA synthesis

text: Synthesis of the negative‐strand ((−)‐strand) counterpart is the first step of coronavirus (CoV) replication; however, the detailed mechanism of the early event and the factors involved remain to be determined. Here, using bovine coronavirus (BCoV)‐defective interfering (DI) RNA, we showed that (a) a poly(A) tail with a length of 15 nucleotides (nt) was sufficient to initiate efficient (−)‐strand RNA synthesis and (b) substitution of the poly(A) tail with poly(U), (C) or (G) only slightly decreased the efficiency of (−)‐strand synthesis. The findings indicate that in addition to the poly(A) tail, other factors acting in trans may also participate in (−)‐strand synthesis. The BCoV nucleocapsid (N) protein, an RNA‐binding protein, was therefore tested as a candidate. Based on dissociation constant (K(d)) values, it was found that the binding affinity between N protein, but not poly(A)‐binding protein, and the 3′‐terminal 55 nt plus a poly(A), poly(U), poly(C) or poly(G) tail correlates with the efficiency of (−)‐strand synthesis. Such an association was also evidenced by the binding affinity between the N protein and 5′‐ and 3′‐terminal cis‐acting elements important for (−)‐strand synthesis. Further analysis demonstrated that N protein can act as a bridge to facilitate interaction between the 5′‐ and 3′‐ends of the CoV genome, leading to circularization of the genome. Together, the current study extends our understanding of the mechanism of CoV (−)‐strand RNA synthesis through involvement of N protein and genome circularization and thus may explain why the addition of N protein in trans is required for efficient CoV replication.


title: A Conceptual Framework to Untangle the Concept of Urban Ecosystem Services

text: Abstract Urban ecosystem service (UES) is becoming an influential concept to guide the planning, design, and management of urban landscapes towards urban sustainability. However, its use is hindered by definitional ambiguity, and the conceptual bases underpinning its application remain weak. This is exemplified by two different but equally valid interpretations of UES: “urban ecosystem services”, referring to ecosystem services from analogs of natural and semi-natural ecosystems within urban boundaries, and “urban ecosystem services”, a much broader term that includes the former group as well as urban services in a city. While we recognize that a single definition of UES is not possible nor necessary as its application is context-dependent, it is nevertheless useful to clarify the relationships between these interpretations to promote consistent use, and importantly, explore how a broader interpretation of UES might advance its applications in areas that have been neglected. We developed a conceptual framework that links UES to natural and human-derived capital to explain the relationships between the dual meanings of UES and proposed three normative propositions to guide its application: (1) integrate holistically multiple components of natural capital to provide UES, (2) reduce dependence on non-renewable abiotic resources and human-derived capital, and (3) enhance UES through technology. The framework we developed helps to resolve the current ambiguity in the meanings of UES, highlights the need to recognise neglected aspects of natural capital important for UES, and can be used to clarify relationships with related concepts conveying dependence of human well-being on nature.


title: Two Novel Parvoviruses in Frugivorous New and Old World Bats

text: Bats, a globally distributed group of mammals with high ecological importance, are increasingly recognized as natural reservoir hosts for viral agents of significance to human and animal health. In the present study, we evaluated pools of blood samples obtained from two phylogenetically distant bat families, in particular from flying foxes (Pteropodidae), Eidolon helvum in West Africa, and from two species of New World leaf-nosed fruit bats (Phyllostomidae), Artibeus jamaicensis and Artibeus lituratus in Central America. A sequence-independent virus discovery technique (VIDISCA) was used in combination with high throughput sequencing to detect two novel parvoviruses: a PARV4-like virus named Eh-BtPV-1 in Eidolon helvum from Ghana and the first member of a putative new genus in Artibeus jamaicensis from Panama (Aj-BtPV-1). Those viruses were circulating in the corresponding bat colony at rates of 7–8%. Aj-BtPV-1 was also found in Artibeus lituratus (5.5%). Both viruses were detected in the blood of infected animals at high concentrations: up to 10E8 and to 10E10 copies/ml for Aj-BtPV-1 and Eh-BtPV-1 respectively. Eh-BtPV-1 was additionally detected in all organs collected from bats (brain, lungs, liver, spleen, kidneys and intestine) and spleen and kidneys were identified as the most likely sites where viral replication takes place. Our study shows that bat parvoviruses share common ancestors with known parvoviruses of humans and livestock. We also provide evidence that a variety of Parvovirinae are able to cause active infection in bats and that they are widely distributed in these animals with different geographic origin, ecologies and climatic ranges.


Drug reaction with eosinophilia and systemic symptoms syndrome to hydroxychloroquine, an old drug in the spotlight in the COVID‐19 era


title: The impact of COVID-19 partial lockdown on the air quality of the city of Rio de Janeiro, Brazil

text: Abstract The first COVID-19 case in Brazil was confirmed on February 25, 2020. On March 16, the state's governor declared public health emergency in the city of Rio de Janeiro and partial lockdown measures came into force a week later. The main goal of this work is to discuss the impact of the measures on the air quality of the city by comparing the particulate matter, carbon monoxide, nitrogen dioxide and ozone concentrations determined during the partial lockdown with values obtained in the same period of 2019 and also with the weeks prior to the virus outbreak. Concentrations varied with substantial differences among pollutants and also among the three studied monitoring stations. CO levels showed the most significant reductions (30.3–48.5%) since they were related to light-duty vehicular emissions. NO2 also showed reductions while PM10 levels were only reduced in the first lockdown week. In April, an increase in vehicular flux and movement of people was observed mainly as a consequence of the lack of consensus about the importance and need of social distancing and lockdown. Ozone concentrations increased probably due to the decrease in nitrogen oxides level. When comparing with the same period of 2019, NO2 and CO median values were 24.1–32.9 and 37.0–43.6% lower. Meteorological interferences, mainly the transport of pollutants from the industrial areas might have also impacted the results.


title: Phylogenetic evidence of a novel lineage of canine pneumovirus and a naturally recombinant strain isolated from dogs with respiratory illness in Thailand

text: BACKGROUND: Canine pneumovirus (CPV) is a pathogen that causes respiratory disease in dogs, and recent outbreaks in shelters in America and Europe have been reported. However, based on published data and documents, the identification of CPV and its variant in clinically symptomatic individual dogs in Thailand through Asia is limited. Therefore, the aims of this study were to determine the emergence of CPV and to consequently establish the genetic characterization and phylogenetic analysis of the CPV strains from 209 dogs showing respiratory distress in Thailand. RESULTS: This study identified and described the full-length CPV genome from three strains, designated herein as CPV_CP13 TH/2015, CPV_CP82 TH/2016 and CPV_SR1 TH/2016, that were isolated from six dogs out of 209 dogs (2.9%) with respiratory illness in Thailand. Phylogenetic analysis suggested that these three Thai CPV strains (CPV TH strains) belong to the CPV subgroup A and form a novel lineage; proposed as the Asian prototype. Specific mutations in the deduced amino acids of these CPV TH strains were found in the G/glycoprotein sequence, suggesting potential substitution sites for subtype classification. Results of intragenic recombination analysis revealed that CPV_CP82 TH/2016 is a recombinant strain, where the recombination event occurred in the L gene with the Italian prototype CPV Bari/100–12 as the putative major parent. Selective pressure analysis demonstrated that the majority of the nucleotides in the G/glycoprotein were under purifying selection with evidence of positive selection sites. CONCLUSIONS: This collective information on the CPV TH strains is the first evidence of CPV emergence with genetic characterization in Thailand and as first report in Asia, where homologous recombination acts as a potential force driving the genetic diversity and shaping the evolution of canine pneumovirus. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-019-2035-1) contains supplementary material, which is available to authorized users.


title: Production and immunogenicity of chimeric virus-like particles containing the spike glycoprotein of infectious bronchitis virus

text: Infectious bronchitis virus (IBV) poses a severe threat to the poultry industry and causes heavy economic losses worldwide. Vaccination is the most effective method of preventing infection and controlling the spread of IBV, but currently available inactivated and attenuated virus vaccines have some disadvantages. We developed a chimeric virus-like particle (VLP)-based candidate vaccine for IBV protection. The chimeric VLP was composed of matrix 1 protein from avian influenza H5N1 virus and a fusion protein neuraminidase (NA)/spike 1 (S1) that was generated by fusing IBV S1 protein to the cytoplasmic and transmembrane domains of NA protein of avian influenza H5N1 virus. The chimeric VLPs elicited significantly higher S1-specific antibody responses in intramuscularly immunized mice and chickens than inactivated IBV viruses. Furthermore, the chimeric VLPs induced significantly higher neutralization antibody levels than inactivated H120 virus in SPF chickens. Finally, the chimeric VLPs induced significantly higher IL-4 production in mice. These results demonstrate that chimeric VLPs have the potential for use in vaccines against IBV infection.


Third Director of the European Centre for Disease Prevention and Control takes office