title: Expression, post‐translational modification and biochemical characterization of proteins encoded by subgenomic mRNA8 of the severe acute respiratory syndrome coronavirus

text: The most striking difference between the subgenomic mRNA8 of severe acute respiratory syndrome coronavirus isolated from human and some animal species is the deletion of 29 nucleotides, resulting in splitting of a single ORF (ORF8) into two ORFs (ORF8a and ORF8b). ORF8a and ORF8b are predicted to encode two small proteins, 8a and 8b, and ORF8 a single protein, 8ab (a fusion form of 8a and 8b). To understand the functions of these proteins, we cloned cDNA fragments covering these ORFs into expression plasmids, and expressed the constructs in both in vitro and in vivo systems. Expression of a construct containing ORF8a and ORF8b generated only a single protein, 8a; no 8b protein expression was obtained. Expression of a construct containing ORF8 generated the 8ab fusion protein. Site‐directed mutagenesis and enzymatic treatment revealed that protein 8ab is modified by N‐linked glycosylation on the N81 residue and by ubiquitination. In the absence of the 8a region, protein 8b undergoes rapid degradation by proteasomes, and addition of proteasome inhibitors inhibits the degradation of protein 8b as well as the protein 8b‐induced rapid degradation of the severe acute respiratory syndrome coronavirus E protein. Glycosylation could also stabilize protein 8ab. More interestingly, the two proteins could bind to monoubiquitin and polyubiquitin, suggesting the potential involvement of these proteins in the pathogenesis of severe acute respiratory syndrome coronavirus.


title: High Contagiousness and Rapid Spread of Severe Acute Respiratory Syndrome Coronavirus 2

text: Severe acute respiratory syndrome coronavirus 2 is the causative agent of the ongoing coronavirus disease pandemic. Initial estimates of the early dynamics of the outbreak in Wuhan, China, suggested a doubling time of the number of infected persons of 6–7 days and a basic reproductive number (R(0)) of 2.2–2.7. We collected extensive individual case reports across China and estimated key epidemiologic parameters, including the incubation period (4.2 days). We then designed 2 mathematical modeling approaches to infer the outbreak dynamics in Wuhan by using high-resolution domestic travel and infection data. Results show that the doubling time early in the epidemic in Wuhan was 2.3–3.3 days. Assuming a serial interval of 6–9 days, we calculated a median R(0) value of 5.7 (95% CI 3.8–8.9). We further show that active surveillance, contact tracing, quarantine, and early strong social distancing efforts are needed to stop transmission of the virus.


title: Sample Pretreatment and Nucleic Acid-Based Detection for Fast Diagnosis Utilizing Microfluidic Systems

text: Recently, micro-electro-mechanical-systems (MEMS) technology and micromachining techniques have enabled miniaturization of biomedical devices and systems. Not only do these techniques facilitate the development of miniaturized instrumentation for biomedical analysis, but they also open a new era for integration of microdevices for performing accurate and sensitive diagnostic assays. A so-called “micro-total-analysis-system”, which integrates sample pretreatment, transport, reaction, and detection on a small chip in an automatic format, can be realized by combining functional microfluidic components manufactured by specific MEMS technologies. Among the promising applications using microfluidic technologies, nucleic acid-based detection has shown considerable potential recently. For instance, micro-polymerase chain reaction chips for rapid DNA amplification have attracted considerable interest. In addition, microfluidic devices for rapid sample pretreatment prior to nucleic acid-based detection have also achieved significant progress in the recent years. In this review paper, microfluidic systems for sample preparation, nucleic acid amplification and detection for fast diagnosis will be reviewed. These microfluidic devices and systems have several advantages over their large-scale counterparts, including lower sample/reagent consumption, lower power consumption, compact size, faster analysis, and lower per unit cost. The development of these microfluidic devices and systems may provide a revolutionary platform technology for fast sample pretreatment and accurate, sensitive diagnosis.


SARS‐CoV‐2 infection in two pediatric patients with immune cytopenias: A single institution experience during the pandemic


title: Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization

text: We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10(0) copies/µL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.


title: Pandemics and Psychiatry: repositioning research in context of COVID-19

text: The current Covid-19 pandemic is posing an unprecedented threat to global well-being and its mental health consequences are just becoming understood. While the crisis has led to a temporary halt in a lot of ongoing or planned psychiatric research, this pandemic is a natural experiment that can help the field to repurpose research to better understand the causes, presentations and outcome trajectories as well as treatments in psychiatry. Inter-disciplinary collaborations between researchers are needed to rapidly develop and share the emerging new knowledge of the mental health implications of Covid-19, This will help mount an effective response to the current as well as future pandemics.


title: One-step multiplex TaqMan probe-based method for real-time PCR detection of four canine diarrhea viruses

text: Viral canine diarrhea has high morbidity and mortality and is prevalent worldwide, resulting in severe economic and spiritual losses to pet owners. However, diarrhea pathogens have similar clinical symptoms and are difficult to diagnose clinically. Thus, fast and accurate diagnostic methods are of great significance for prevention and accurate treatment. In this study, we developed a one-step multiplex TaqMan probe-based real-time PCR for the differential diagnosis of four viruses causing canine diarrhea including, CPV (Canine Parvovirus 2), CCoV (Canine Coronavirus), CAstV (Canine Astrovirus), and CaKoV (Canine Kobuviruses). The limit of detection was up to 10(2)copies/μL and performed well with high sensitivity and specificity. This assay was optimized and used to identify possible antagonistic relationships between viruses. From this, artificial pre-experiments were performed for mixed infections, and a total of 82 canine diarrhea field samples were collected from different animal hospitals in Zhejiang, China to assess the method. The virus prevalence was significantly higher than what previously reported based on RT-PCR(Reverse Transcription-Polymerase Chain Reaction). Taken together, these results suggest that the method can be used as a preferred tool for monitoring laboratory epidemics, timely prevention, and effective monitoring of disease progression.


docid: dpwzgake

title: Plants as bioreactors for the production of vaccine antigens

text: Abstract Plants have been identified as promising expression systems for commercial production of vaccine antigens. In phase I clinical trials several plant-derived vaccine antigens have been found to be safe and induce sufficiently high immune response. Thus, transgenic plants, including edible plant parts are suggested as excellent alternatives for the production of vaccines and economic scale-up through cultivation. Improved understanding of plant molecular biology and consequent refinement in the genetic engineering techniques have led to designing approaches for high level expression of vaccine antigens in plants. During the last decade, several efficient plant-based expression systems have been examined and more than 100 recombinant proteins including plant-derived vaccine antigens have been expressed in different plant tissues. Estimates suggest that it may become possible to obtain antigen sufficient for vaccinating millions of individuals from one acre crop by expressing the antigen in seeds of an edible legume, like peanut or soybean. In the near future, a plethora of protein products, developed through ‘naturalized bioreactors’ may reach market. Efforts for further improvements in these technologies need to be directed mainly towards validation and applicability of plant-based standardized mucosal and edible vaccines, regulatory pharmacology, formulations and the development of commercially viable GLP protocols. This article reviews the current status of developments in the area of use of plants for the development of vaccine antigens.


title: A Collaborative Multidisciplinary Approach to the Management of Coronavirus Disease-19 in the Hospital Setting

text: • Supportive care is the standard approach to the management of coronavirus disease-19, with particular attention to respiratory support, and early detection of potential complications such as cytokine storm and organ dysfunction. • Therapeutic management of coronavirus disease-19 using antivirals and immunomodulators remain investigational, and should be implemented using clinical trial protocols. • 3. A multidisciplinary collaborative approach to management of coronavirus disease-19 is an essential component of providing optimal care to patients.


docid: oqvsiawe

title: Bronchiolitis Obliterans Organizing Pneumonia in Swine Associated with Porcine Circovirus Type 2 Infection

text: Bronchiolitis obliterans organizing pneumonia (BOOP) is a chronic respiratory disease. Although the pathogenesis of BOOP is still incompletely understood, BOOP is responsive to steroids and has a good prognosis. In our five pigs with chronic postweaning multisystemic wasting syndrome (PMWS), typical BOOP lesions were revealed. All five porcine lungs showed typical intraluminal plugs, and porcine circovirus type 2 (PCV2) was identified. They also exhibited similar pathologic findings such as proliferation of type II pneumocytes and myofibroblasts (MFBs), extracellular collagen matrix (ECM) deposition, and fragmentation of elastic fibers. MFBs migration correlative molecules, for instance, gelatinase A, B and osteopontin, appeared strongly in the progressing marginal area of polypoid intraluminal plugs of fibrotic lesion. These molecules colocalized with the active MFBs. Both gelatinase activity and intercellular level of active MFBs were significantly increased (P < .05). Porcine chronic bronchopneumonia leads to BOOP and it is associated with PCV2 persistent infection. Swine BOOP demonstrates similar cellular constituents with human BOOP. Perhaps their molecular mechanisms of pathogenesis operate in a similar way. Thus we infer that the swine BOOP can be considered as a potential animal model for human BOOP associated with natural viral infection. Moreover, it is more convenient to obtain samples.


title: Feline vaccines

text: Many of the decisions regarding the vaccination of cats relate to the animal’s lifestyle. Vaccination requirements for the solitary indoor cat are very different than those for feral or free-roaming cats. Core vaccines include those directed against feline herpesvirus, feline parvovirus, and feline caliciviruses. Other important vaccines include the mandated rabies vaccination and also vaccination against feline leukemia. One significant issue with respect to feline vaccination is the development of injection site sarcomas. Although the prevalence of these is low and should not inhibit the use of vaccines, they are impossible to predict and very difficult to treat.


title: Two Genetically Similar H9N2 Influenza A Viruses Show Different Pathogenicity in Mice

text: H9N2 Avian influenza virus has repeatedly infected humans and other mammals, which highlights the need to determine the pathogenicity and the corresponding mechanism of this virus for mammals. In this study, we found two H9N2 viruses with similar genetic background but with different pathogenicity in mice. The A/duck/Nanjing/06/2003 (NJ06) virus was highly pathogenic for mice, with a 50% mouse lethal dose (MLD(50)) of 10(2.83) 50% egg infectious dose (EID(50)), whereas the A/duck/Nanjing/01/1999 (NJ01) virus was low pathogenic for mice, with a MLD(50) of >10(6.81) EID(50). Further studies showed that the NJ06 virus grew faster and reached significantly higher titers than NJ01 in vivo and in vitro. Moreover, the NJ06 virus induced more severe lung lesions, and higher levels of inflammatory cellular infiltration and cytokine response in lungs than NJ01 did. However, only 12 different amino acid residues (HA-K157E, NA-A9T, NA-R435K, PB2-T149P, PB2-K627E, PB1-R187K, PA-L548M, PA-M550L, NP-G127E, NP-P277H, NP-D340N, NS1-D171N) were found between the two viruses, and all these residues except for NA-R435K were located in the known functional regions involved in interaction of viral proteins or between the virus and host factors. Summary, our results suggest that multiple amino acid differences may be responsible for the higher pathogenicity of the NJ06 virus for mice, resulting in lethal infection, enhanced viral replication, severe lung lesions, and excessive inflammatory cellular infiltration and cytokine response in lungs. These observations will be helpful for better understanding the pathogenic potential and the corresponding molecular basis of H9N2 viruses that might pose threats to human health in the future.


Screening Laboratory Requests


title: Recombinant Vesicular Stomatitis Virus Vaccine Vectors Expressing Filovirus Glycoproteins Lack Neurovirulence in Nonhuman Primates

text: The filoviruses, Marburg virus and Ebola virus, cause severe hemorrhagic fever with high mortality in humans and nonhuman primates. Among the most promising filovirus vaccines under development is a system based on recombinant vesicular stomatitis virus (rVSV) that expresses an individual filovirus glycoprotein (GP) in place of the VSV glycoprotein (G). The main concern with all replication-competent vaccines, including the rVSV filovirus GP vectors, is their safety. To address this concern, we performed a neurovirulence study using 21 cynomolgus macaques where the vaccines were administered intrathalamically. Seven animals received a rVSV vector expressing the Zaire ebolavirus (ZEBOV) GP; seven animals received a rVSV vector expressing the Lake Victoria marburgvirus (MARV) GP; three animals received rVSV-wild type (wt) vector, and four animals received vehicle control. Two of three animals given rVSV-wt showed severe neurological symptoms whereas animals receiving vehicle control, rVSV-ZEBOV-GP, or rVSV-MARV-GP did not develop these symptoms. Histological analysis revealed major lesions in neural tissues of all three rVSV-wt animals; however, no significant lesions were observed in any animals from the filovirus vaccine or vehicle control groups. These data strongly suggest that rVSV filovirus GP vaccine vectors lack the neurovirulence properties associated with the rVSV-wt parent vector and support their further development as a vaccine platform for human use.


Potential Threats of COVID-19 On Quarantined Families


title: A serological comparison of bovine coronavirus strains

text: Two bovine coronavirus (BCV) strains from diarrheic calf faeces were adapted to grow in HRT 18 cells and compared in immunofluorescence (IF), haemagglutination inhibition (HAI) and neutralisation (NT) tests with three other strains of BCV and a human coronavirus (HCV) strain obtained from other laboratories. Polyclonal antisera against these 6 viruses were raised in rabbits. No significant differences between viruses were detected by IF. In the HAI test the HCV strain was distinguishable from the 5 BCV strains and serological variation between the BCV strains was shown. HCV could be distinguished by NT test, but all BCV isolates were similar. Two monoclonal antibodies prepared against one of the BCV strains distinguished HCV in all three assays, and detected varying relationships between BCV strains.


title: Characterizing the Impact of COVID-19 on Men Who Have Sex with Men Across the United States in April, 2020

text: The COVID-19 pandemic is reinforcing health inequities among vulnerable populations, including men who have sex with men (MSM). We conducted a rapid online survey (April 2 to April 13, 2020) of COVID-19 related impacts on the sexual health of 1051 US MSM. Many participants had adverse impacts to general wellbeing, social interactions, money, food, drug use and alcohol consumption. Half had fewer sex partners and most had no change in condom access or use. Some reported challenges in accessing HIV testing, prevention and treatment services. Compared to older MSM, those 15–24 years were more likely to report economic and service impacts. While additional studies of COVID-19 epidemiology among MSM are needed, there is already evidence of emerging interruptions to HIV-related services. Scalable remote solutions such as telehealth and mailed testing and prevention supplies may be urgently needed to avert increased HIV incidence among MSM during the COVID-19 pandemic era. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10461-020-02894-2) contains supplementary material, which is available to authorized users.


Authors’ response: Plenty of coronaviruses but no SARS-CoV-2


title: Testing cardiovascular autonomic function in the COVID-19 era: lessons from Bologna’s Autonomic Unit

text: The coronavirus disease 2019 (COVID-19) pandemic has changed the way most medical procedures are performed. Autonomic units, as well as other healthcare sectors, are required to undergo a thorough reorganization of the protocols in order to guarantee the safety of patients and healthcare staff. Cardiovascular autonomic function testing (CAFT) is necessary in certain situations; however, it poses several concerns which need to be addressed. Here, we provide some practical advice based on current national and international health authorities’ recommendations and our experience about how to perform CAFT during the COVID-19 emergency. We examine aspects regarding patients, healthcare staff, laboratory preparation, and test performance.


title: SARS coronavirus replicase proteins in pathogenesis

text: Much progress has been made in understanding the role of structural and accessory proteins in the pathogenesis of severe acute respiratory syndrome coronavirus (SARS-CoV) infections. The SARS epidemic also brought new attention to the proteins translated from ORF1a and ORF1b of the input genome RNA, also known as the replicase/transcriptase gene. Evidence for change within the ORF1ab coding sequence during the SARS epidemic, as well as evidence from studies with other coronaviruses, indicates that it is likely that the ORF1ab proteins play roles in virus pathogenesis distinct from or in addition to functions directly involved in viral replication. Recent reverse genetic studies have confirmed that proteins of ORF1ab may be involved in cellular signaling and modification of cellular gene expression, as well as virulence by mechanisms yet to be determined. Thus, the evolution of the ORF1ab proteins may be determined as much by issues of host range and virulence as they are by specific requirements for intracellular replication.


title: Manejo quirúrgico de pacientes con infección por COVID-19. Recomendaciones de la Asociación Española de Cirujanos

text: Due to the current pandemic of respiratory disease known as coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2 virus, many patients with confirmed or suspected COVID-19 infection will require elective surgery, surgery that cannot be postponed, or emergency surgical treatment. In these situations, special measures need to be adopted in order to minimize the possibility of transmission between patients, exposure of healthcare personnel and the development of postoperative complications. This document explains the main principles to consider when managing confirmed or suspected COVID-19 patients during evaluation as well as when surgical treatment is required.


title: Coding potential and sequence conservation of SARS-CoV-2 and related animal viruses

text: Abstract In December 2019, a novel human-infecting coronavirus (SARS-CoV-2) was recognized in China. In a few months, SARS-CoV-2 has caused thousands of disease cases and deaths in several countries. Phylogenetic analyses indicated that SARS-CoV-2 clusters with SARS-CoV in the Sarbecovirus subgenus and viruses related to SARS-CoV-2 were identified from bats and pangolins. Coronaviruses have long and complex genomes with high plasticity in terms of gene content. To date, the coding potential of SARS-CoV-2 remains partially unknown. We thus used available sequences of bat and pangolin viruses to determine the selective events that shaped the genome structure of SARS-CoV-2 and to assess its coding potential. By searching for signals of significantly reduced variability at synonymous sites (dS), we identified six genomic regions, one of these corresponding to the programmed −1 ribosomal frameshift. The most prominent signal of dS reduction was observed within the E gene. A genome-wide analysis of conserved RNA structures indicated that this region harbors a putative functional RNA element that is shared with the SARS-CoV lineage. Additional signals of reduced dS indicated the presence of internal ORFs. Whereas the presence ORF9a (internal to N) was previously proposed by homology with a well characterized protein of SARS-CoV, ORF3h (for hypothetical, within ORF3a) was not previously described. The predicted product of ORF3h has 90% identity with the corresponding predicted product of SARS-CoV and displays features suggestive of a viroporin. Finally, analysis of the putative ORF10 revealed high dN/dS (3.82) in SARS-CoV-2 and related coronaviruses. In the SARS-CoV lineage, the ORF is predicted to encode a truncated protein and is neutrally evolving. These data suggest that ORF10 encodes a functional protein in SARS-CoV-2 and that positive selection is driving its evolution. Experimental analyses will be necessary to validate and characterize the coding and non-coding functional elements we identified.


title: Effects of space flight on DNA mutation and secondary metabolites of licorice (Glycyrrhiza uralensis Fisch.)

text: Licorice (Glycyrrhiza uralensis Fisch.) seeds were flown on a recoverable satellite for 18 days(the average radiation dose in the flight recovery module was 0.102 mGy/d, the distance from flight apogee to earth was 350 km, gravity 10(−6)). After returning to earth, the seeds were germinated and grown to maturity. The parallel ground-based seeds were also planted under the same conditions. The leaves of licorice were used for inter-simple sequence repeat (ISSR) analysis and the two main secondary metabolites in one-year-old roots were analyzed by high performance liquid chromatography (HPLC). Among 22 random primers used in this experiment, 6 primers generated different DNA band types. Analysis of HPLC showed that the content of glycyrrhizic acid (GA) and liquiritin (LQ) in the roots from seeds flown in space was respectively 2.19, 1.18 times higher than that of the control group. The results demonstrated that the extraterrestrial environment induced mutagenic effects on licorice and affected its secondary metabolites. These changes indicated that extraterrestrial orbit is possible means of breeding of licorice so as to preserve this endangered medicinal plant.


Transitioning clinical rotations to a virtual experience: Problem, solution, and results


title: SARS-coronavirus spike S2 domain flanked by cysteine residues C822 and C833 is important for activation of membrane fusion

text: Abstract The S2 domain of the coronavirus spike (S) protein is known to be responsible for mediating membrane fusion. In addition to a well-recognized cleavage site at the S1–S2 boundary, a second proteolytic cleavage site has been identified in the severe acute respiratory syndrome coronavirus (SARS-CoV) S2 domain (R797). C-terminal to this S2 cleavage site is a conserved region flanked by cysteine residues C822 and C833. Here, we investigated the importance of this well conserved region for SARS-CoV S-mediated fusion activation. We show that the residues between C822–C833 are well conserved across all coronaviruses. Mutagenic analysis of SARS-CoV S, combined with cell–cell fusion and pseudotyped virion infectivity assays, showed a critical role for the core-conserved residues C822, D830, L831, and C833. Based on available predictive models, we propose that the conserved domain flanked by cysteines 822 and 833 forms a loop structure that interacts with components of the SARS-CoV S trimer to control the activation of membrane fusion.


Expression and Characterization of Recombinant S2 Subunit of SARS-coronavirus S Fusion Protein


title: Identification of an infectious bronchitis coronavirus strain exhibiting a classical genotype but altered antigenicity, pathogenicity, and innate immunity profile

text: Avian coronavirus infectious bronchitis virus (IBV) poses economic threat to the poultry industry worldwide. Pathogenic IBV 3575/08 was isolated from broilers vaccinated with the attenuated viral vaccine derived from a Taiwan strain 2575/98. In this study, extensive investigations were conducted on the genome sequences, antigenicity, pathogenicity, and host immune responses of several IBV strains in specific-pathogen-free chickens. Sequence analyses revealed that 3575/08 and 2575/98 shared high homology in their structural genes, but not in non-structural accessory proteins such as 3a, 3b and 5b. Despite a high degree of homology in their spike protein genes, cross neutralization test showed low cross protection between 3575/08 and 2575/98, suggesting distinct antigenicity for the two strains. Animal challenge experiments exhibited strong respiratory and renal pathogenicity for 3575/08. In addition, early and prolonged viral shedding and rapid viral dissemination were observed. Immune gene expression profiling by PCR array showed chickens infected with 3575/08 had delayed expression of a subset of early innate immune genes, whereas chickens infected with the wild-type or attenuated-type 2575/08 revealed quick gene induction and efficient virus control. In summary, this study reveals a new IBV strain, which harbors a known local genotype but displays remarkably altered antigenicity, pathogenicity and host defenses.


title: Missed opportunities for antimicrobial stewardship in pre‐school children admitted to hospital with lower respiratory tract infection

text: AIM: To describe the usage of multiplex polymerase chain reaction on nasopharyngeal swab (NPS) samples in pre‐school children presenting with lower respiratory tract infection (LRTI) at Christchurch Hospital, and its impact on the use of antibiotics empirically and at discharge. METHODS: This retrospective cohort study included 237 children, ages 3 months to 5 years, admitted to hospital during the winter months of 2012–2015 with a diagnosis of community‐acquired LRTI. Children were identified by discharge coding and their notes reviewed. RESULTS: A significantly larger proportion of children who had a NPS sample taken (42/146, 36%) received no empiric antibiotics compared with children who did not have a sample taken (7/91, 7.7%, P < 0.001). Of those who did have a NPS sample taken 17 of 146 (11.6%) had their antibiotics discontinued prior to or at the time of discharge compared with only 3 of 91 (3.3%) of those who did not have a NPS sample (P < 0.025). Children with influenza detected were more likely to receive no antibiotics or have their antibiotics discontinued prior to or at discharge. Only a small proportion of children with other viruses identified had their antibiotics discontinued. CONCLUSIONS: It appears that clinicians were generally reluctant to stop antibiotics prior to discharge in young children with LRTI in whom influenza or other viruses were identified. In our view, it makes sense to stop antibiotics when the clinical presentation and NPS testing is consistent with a viral aetiology. Not stopping antibiotics at or before discharge in these children represents a missed opportunity for antimicrobial stewardship.


title: Contagion

text: Are we really a few mutations away from the end of the world as we know it? Emma Young investigates


title: Autoinflammatory and autoimmune conditions at the crossroad of COVID-19

text: Coronavirus disease 2019 (COVID-19) has been categorized as evolving in overlapping phases. First, there is a viral phase that may well be asymptomatic or mild in the majority, perhaps 80% of patients. The pathophysiological mechanisms resulting in minimal disease in this initial phase are not well known. In the remaining 20% of cases, the disease may become severe and/or critical. In most patients of this latter group, there is a phase characterized by the hyperresponsiveness of the immune system. A third phase corresponds to a state of hypercoagulability. Finally, in the fourth stage organ injury and failure occur. Appearance of autoinflammatory/autoimmune phenomena in patients with COVID-19 calls attention for the development of new strategies for the management of life-threatening conditions in critically ill patients. Antiphospholipid syndrome, autoimmune cytopenia, Guillain-Barré syndrome and Kawasaki disease have each been reported in patients with COVID-19. Here we present a scoping review of the relevant immunological findings in COVID-19 as well as the current reports about autoimmune/autoinflammatory conditions associated with the disease. These observations have crucial therapeutic implications since immunomodulatory drugs are at present the most likely best candidates for COVID-19 therapy. Clinicians should be aware of these conditions in patients with COVID-19, and these observations should be considered in the current development of vaccines.


title: Griffithsin inhibits porcine reproductive and respiratory syndrome virus infection in vitro

text: Porcine reproductive and respiratory syndrome virus (PRRSV) is a pathogen that severely disrupts swine production. Despite sustained efforts, the disease is still endemic, with high mortality and morbidity. New antiviral strategies to control PRRSV are needed. Griffithsin, a red algal lectin, has potent antiviral effect on several human enveloped viruses, but this effect has not been demonstrated on PRRSV. Here, we first tested the in vitro antiviral activity of Griffithsin against PRRSV. Griffithsin exerted strong saccharide-dependent antiviral activity against PRRSV, probably through interactions with glycans on the surface of PRRSV that interfered with virus entry. Furthermore we revealed that Griffithsin’s action on PRRSV involved blocking viral adsorption, and it had no effect on viral penetration. Besides Our findings also suggested that Griffithsin may interfere with cell-to-cell spread to prevent virus transmission. The remarkable potency profile of Griffithsin supports its potential value as an antiviral agent against PRRSV.


title: Demand‐capacity modelling and Covid‐19 disease: identifying themes for future NHS planning

text: This editorial shows how the principles of demand‐capacity modelling lead to the conclusion that COVID‐19 disease will require the UK NHS to run its serves at a far higher working capacity (in terms of hospital, intensive care and social care beds, operating theatres and staff) than it has been used to. The analysis leads to key questions that should form the basis for rational planning in a post‐COVID NHS.


title: Verification with the utility of an established rapid assessment of brain safety for newly developed vaccines

text: In the twenty-first century, high contagious infectious diseases such as SARS (Severe Acute Respiratory Syndrome), MERS (Middle East Respiratory Syndrome), FMD (Foot-and-Mouth Disease) and AI (Avian Influenza) have become very prevalent, causing treat harm to humans and animals in aspect of public health, and economical issues. The critical problem is that newly-reported infectious diseases that humans firstly experience are expected to continue to emerge, and these diseases will be spreading out rapidly. Therefore, rapid and safe supplies of effective vaccines are most pivotal to prevent the rapid prevalent of new infection, but international standards or assessing protocol the safety of urgent vaccines are not established well. In our previous study, since we established a module to assess the brain safety of urgent vaccines, therefore, it is necessary to verify that this established module for assessing brain safety could work effectively in commercially available two vaccines (one killed- and on live-vaccines). We compared the results of Evans blue (EB) assay and qPCR analysis by injection of two kinds of vaccines, PBS and Lipopolysaccharide (LPS) under the condition of the module previously reported. We confirmed that the brain safety test module for urgent vaccine we established is very reproducible. Therefore, it is believed that this vaccine safety testing method can be used to validate brain safety when prompt supply of a newly developed vaccines is needed.


title: Human Betacoronavirus 2c EMC/2012–related Viruses in Bats, Ghana and Europe

text: We screened fecal specimens of 4,758 bats from Ghana and 272 bats from 4 European countries for betacoronaviruses. Viruses related to the novel human betacoronavirus EMC/2012 were detected in 46 (24.9%) of 185 Nycteris bats and 40 (14.7%) of 272 Pipistrellus bats. Their genetic relatedness indicated EMC/2012 originated from bats.


title: Least cost energy system pathways towards 100% renewable energy in Ireland by 2050

text: Studies focusing on 100% renewable energy systems have emerged in recent years; however, existing studies tend to focus only on the power sector using exploratory approaches. This paper therefore undertakes a whole-system approach and explores optimal pathways towards 100% renewable energy by 2050. The analysis is carried out for Ireland, which currently has the highest share of variable renewable electricity on a synchronous power system. Large numbers of scenarios are developed using the Irish TIMES model to address uncertainties. Results show that compared to decarbonization targets, focusing on renewable penetration without considering carbon capture options is significantly less cost effective in carbon mitigation. Alternative assumptions on bioenergy imports and maximum variability in power generation lead to very different energy mixes in bioenergy and electrification levels. All pathways suggest that indigenous bioenergy needs to be fully exploited and the current annual deployment rate of renewable electricity needs a boost. Pathways relying on international bioenergy imports are slightly cheaper and faces less economic and technical challenges. However, given the large future uncertainties, it is recommended that further policy considerations be given to pathways with high electrification levels as they are more robust towards uncertainties.


title: An enzymatic assay based on luciferase Ebola virus-like particles for evaluation of virolytic activity of antimicrobial peptides

text: Antimicrobial peptides are currently considered as promising antiviral compounds. Current assays to evaluate the effectivity of peptides against enveloped viruses based on liposomes or hemolysis are encumbered by the artificial nature of liposomes or distinctive membrane composition of used erythrocytes. We propose a novel assay system based on enzymatic Ebola virus-like particles containing sensitive luciferase reporter. The assay was validated with several cationic and anionic peptides and compared with lentivirus inactivation and hemolytic assays. The assay is sensitive and easy to perform in standard biosafety level laboratory with potential for high-throughput screens. The use of virus-like particles in the assay provides a system as closely related to the native viruses as possible eliminating some issues associated with other more artificial set ups. We have identified CAM-W (KWKLWKKIEKWGQGIGAVLKWLTTWL) as a peptide with the greatest antiviral activity against infectious lentiviral vectors and filoviral virus-like particles.


title: Epidemiologic investigation of a family cluster of imported ZIKV cases in Guangdong, China: probable human-to-human transmission

text: Zika virus (ZIKV) is an emerging mosquito-borne flavivirus that can potentially threaten South China. A Chinese family of four returning from Venezuela to China was found to be positive for ZIKV when the youngest son's fever was first detected at an airport immigration inspection. They were isolated temporarily in a local hospital in Enping city, Guangdong province, where their clinical data were recorded and urine and saliva were collected to isolate ZIKV and to obtain viral sequences. All of them except the mother presented mild symptoms of rash and fever. Envelope gene sequences from the father, daughter and son were completely identical. Phylogenetic analysis demonstrated that this strain is similar to several imported strains reported in recent months, which are all clustered into a group isolated from 2015 ZIKA outbreaks in Brazil. Together with the climatic features in Venezuela, New York and Guangdong in February, it can be concluded that our subjects are imported cases from Venezuela. With the same viral sequence being shared between family members, neither direct human-to-human nor vector transmission can be ruled out in this study, but the former seems more likely. Although our subjects had mild illness, epidemiologists and public health officials should be aware of the risk of further expansion of ZIKV transmission by local competent vectors.


title: Acute Cough in the Elderly: Aetiology, Diagnosis and Therapy

text: Although the frequency of physician consultations and the sale of over-the-counter remedies establish the high prevalence of acute cough in the elderly, epidemiological studies have tended to be imprecise. However, respiratory tract infections in nose, larynx and/or bronchi, either viral or bacterial or both, are by far the commonest cause of acute cough. These are especially frequent and hazardous in the elderly, and community living and institutionalisation may aggravate this problem. A variety of viruses and bacteria have been incriminated, with rhinovirus, influenza and respiratory syncytial viruses, and Streptococcus pneumoniae, Haemophilus influenza and Bordetella pertussis being especially important. Viral infections can readily lead to community-acquired pneumonia. Successful diagnosis should point to successful treatment, and in this respect clinical examination and patient history are paramount, supplemented by chest X-ray, viral and bacterial culture and serological testing. Depending on the results of these tests, specific antibacterial therapy may be called for, although there is dispute as to the merits of antibacterial therapy in cases of uncertain diagnosis. Prevention and prophylaxis for influenza and S. pneumoniae infections are now commendably routine in the elderly, especially those in communities. Treatment, as well as the use of antibacterials, may also be directed against the inflammatory and infective processes in the airways. Non-specific antitussive therapy is common and usually highly desirable to prevent the adverse effects of repeated coughing. There have been few advances in antitussive therapy in recent years, opioids and dextromethorphan being the most commonly used agents; they act centrally on the brainstem, but also have a large placebo effect. However they work, they are much appreciated by patients and their partners. Moreover, striking advances in our understanding of the peripheral sensory and central nervous pathways of the cough reflex in recent years should soon lead to a new and more specific choice of agents to inhibit cough.


A NEW ANTIRHINOVIRUS COMPOUND, ICI 73602: STRUCTURE, PROPERTIES, AND SPECTRUM OF ACTIVITY


COVID-19: a hypothesis regarding the ventilation-perfusion mismatch


title: Real-Time Fluorescence Loop Mediated Isothermal Amplification for the Detection of Acinetobacter baumannii

text: BACKGROUND: Detection of Acinetobacter baumannii has been relying primarily on bacterial culture that often fails to return useful results in time. Although DNA-based assays are more sensitive than bacterial culture in detecting the pathogen, the molecular results are often inconsistent and challenged by doubts on false positives, such as those due to system- and environment-derived contaminations. In addition, these molecular tools require expensive laboratory instruments. Therefore, establishing molecular tools for field use require simpler molecular platforms. The loop-mediated isothermal amplification method is relatively simple and can be improved for better use in a routine clinical bacteriology laboratory. A simple and portable device capable of performing both the amplification and detection (by fluorescence) of LAMP in the same platform has been developed in recent years. This method is referred to as real-time loop-mediated isothermal amplification. In this study, we attempted to utilize this method for rapid detection of A. baumannii. METHODOLOGY AND SIGNIFICANT FINDINGS: Species-specific primers were designed to test the utility of this method. Clinical samples of A. baumannii were used to determine the sensitivity and specificity of this system compared to bacterial culture and a polymerase chain reaction method. All positive samples isolated from sputum were confirmed to be the species of Acinetobacter by 16S rRNA gene sequencing. The RealAmp method was found to be simpler and allowed real-time detection of DNA amplification, and could distinguish A. baumannii from Acinetobacter calcoaceticus and Acinetobacter genomic species 3. DNA was extracted by simple boiling method. Compared to bacterial culture, the sensitivity and specificity of RealAmp in detecting A. baumannii was 98.9% and 75.0%, respectively. CONCLUSION: The RealAmp assay only requires a single unit, and the assay positivity can be verified by visual inspection. Therefore, this assay has great potential of field use as a molecular tool for detection of A. baumannii.


title: Epidemiological study of influenza virus infections in young adult outpatients from Buenos Aires, Argentina

text: Background Influenza virus is the most common cause of influenza‐like illness (ILI) in adults. In Argentina, studies on influenza and other respiratory viruses were performed mostly in pediatric populations. Objectives To determine: (1) the frequency of influenza virus and other common respiratory viruses in adult outpatients with ILI, (2) whether the signs and symptoms predict viral etiology, (3) whether viral diagnosis changes clinical management or infection control measures and (4) to characterize the influenza strains circulating in the community. Population and methods Nasal and pharyngeal swabs from adult outpatients with ILI attending the emergency room during the winter seasons of 2004 and 2005 in Argentina were evaluated by immunofluorescence and RT‐PCR. Results Of 151 samples analyzed, 39 (26%) were influenza A positive, 5 (3·3%) influenza B positive and 4 (2·6%) respiratory syncytial virus positive by immunofluorescence. Two samples (1·3%) were human metapneumovirus positive by RT PCR. Cell culture detected six additional influenza viruses and one adenovirus positive sample. The sensitivity of immunofluorescence for influenza compared with culture was 70%. Symptoms did not predict etiology. Conclusions In this study, 40% of the patients with ILI had a specific viral infection and 83% were influenza viruses. Viral detection was necessary to determine the etiology as signs and symptoms were not different between patients with or without viral infection. Viral diagnosis was important to implement infectious control measures. Circulating influenza strains in this study were similar to the correspondent vaccine strains selected for the Southern hemisphere.


title: Poxvirus protein evolution: Family-wide assessment of possible horizontal gene transfer events

text: To investigate the evolutionary origins of proteins encoded by the Poxviridae family of viruses, we examined all poxvirus protein coding genes using a method of characterizing and visualizing the similarity between these proteins and taxonomic subsets of proteins in GenBank. Our analysis divides poxvirus proteins into categories based on their relative degree of similarity to two different taxonomic subsets of proteins such as all eukaryote vs. all virus (except poxvirus) proteins. As an example, this allows us to identify, based on high similarity to only eukaryote proteins, poxvirus proteins that may have been obtained by horizontal transfer from their hosts. Although this method alone does not definitively prove horizontal gene transfer, it allows us to provide an assessment of the possibility of horizontal gene transfer for every poxvirus protein. Potential candidates can then be individually studied in more detail during subsequent investigation. Results of our analysis demonstrate that in general, proteins encoded by members of the subfamily Chordopoxvirinae exhibit greater similarity to eukaryote proteins than to proteins of other virus families. In addition, our results reiterate the important role played by host gene capture in poxvirus evolution; highlight the functions of many genes poxviruses share with their hosts; and illustrate which host-like genes are present uniquely in poxviruses and which are also present in other virus families.


title: Antiviral activities of Schizonepeta tenuifolia Briq. against enterovirus 71 in vitro and in vivo

text: No effective drug is currently available for treatment of enterovirus 71 (EV71) infection. Schizonepeta tenuifolia Briq. (ST) has been used as a herbal constituent of traditional Chinese medicine. We studied whether the aqueous extract of Schizonepeta tenuifolia Briq (STE) has antiviral activity. STE inhibited replication of EV71, as evident by its ability to diminish plaque formation and cytopathic effect induced by EV71, and to inhibit the synthesis of viral RNA and protein. Moreover, daily single-dose STE treatment significantly improved the survival of EV71-infected mice, and ameliorated the symptoms. Mechanistically, STE exerts multiple effects on enteroviral infection. Treatment with STE reduced viral attachment and entry; the cleavage of eukaryotic translation initiation factor 4 G (eIF4G) by EV71 protease, 2A(pro); virus-induced reactive oxygen species (ROS) formation; and relocation of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) from the nucleus to the cytoplasm. It was accompanied by a decline in EV71-associated hyperphosphorylation of p38 kinase and EPS15. It is plausible that STE may inhibit ROS-induced p38 kinase activation, and subsequent hnRNP A1 relocation and EPS15-mediated membrane trafficking in infected cells. These findings suggest that STE possesses anti-EV71 activities, and may serve as health food or candidate antiviral drug for protection against EV71.


title: Clinical Management of Pandemic 2009 Influenza A(H1N1) Infection

text: Antiviral therapy and vaccination are important strategies for controlling pandemic 2009 influenza A(H1N1) but efficacy depends on the timing of administration and is often limited by supply shortage. Patients with dyspnea, tachypnea, evidence of hypoxemia, and pulmonary infiltrates on chest radiograph should be hospitalized. Patients with severe illness or underlying medical conditions that increase the risk of more severe disease should be treated with oseltamivir or zanamivir as soon as possible, without waiting for the results of laboratory tests. Lung-protective ventilation strategy with a low tidal volume and adequate pressure, in addition to a conservative fluid management approach, is recommended when treating adult patients with ARDS. Extracorporeal membrane oxygenation has emerged as an important rescue therapy for critically ill patients. Use of systemic steroids was associated with delayed viral clearance in severe acute respiratory syndrome and H3N2 infection. Low-dose corticosteroids may be considered in the treatment of refractory septic shock. Passive immunotherapy in the form of convalescent plasma or hyperimmune globulin may be explored as rescue therapy. More data are needed to explore the potential role of IV gamma globulin and other drugs with immunomodulating properties, such as statins, gemfibrozil, and N-acetyl-cysteine. Health-care workers must apply strict standard and droplet precautions when dealing with suspected and confirmed case and upgrade to airborne precautions when performing aerosol-generating procedures. Nonpharmacologic measures, such as early case isolation, household quarantine, school/workplace closure, good community hygiene, and restrictions on travel are useful measures in controlling an influenza pandemic at its early phase.


title: A Study on Herd Immunity of COVID-19 in South Korea: Using a Stochastic Economic-Epidemiological Model

text: Vaccination is an effective measure to control the diffusion of infectious disease such as COVID-19. This paper analyzes the basic reproduction number in South Korea which enables us to identify a necessary level of vaccine stockpile to achieve herd immunity. An susceptible-infected-susceptible model is adopted that allows a stochastic diffusion. The result shows that the basic reproduction number of South Korea is approximately 2 which is substantially lower than those of the other regions. The herd immunity calculated from economic-epidemiological model suggests that at least 62% of the susceptible population be vaccinated when COVID-19 vaccine becomes available.


title: A DNA prime-protein boost vaccination strategy targeting turkey coronavirus spike protein fragment containing neutralizing epitope against infectious challenge

text: Abstract The present study was undertaken to determine immune response and protection efficacy of a spike (S) protein fragment containing neutralizing epitopes (4F/4R) of turkey coronavirus (TCoV) by priming with DNA vaccine and boosting with the recombinant protein from the corresponding DNA vaccine gene segment. Turkeys were vaccinated by priming with either one dose (G1-750DP) or two doses (G3-750DDP) of 750μg DNA vaccine expressing 4F/4R S fragment and boosting with one dose of 200μg 4F/4R S fragment. One dose of 100μg DNA vaccine mixed with polyethyleneimine (PEI) and sodium hyaluronate (HA) followed by one dose of 750μg DNA vaccine and one dose of 200μg 4F/4R S fragment were given to the turkeys in group G2-100DPH. After infectious challenge by TCoV, clinical signs and TCoV detected by immunofluorescence antibody (IFA) assay were observed in less number of turkeys in vaccination groups than that in challenge control groups. TCoV viral RNA loads measured by quantitative real-time reverse transcription-PCR were lower in vaccinated turkeys than those in challenge control turkeys. The turkeys in G3-750DDP produced the highest level of TCoV S protein-specific antibody and virus neutralization (VN) titer. Comparing to the turkeys in G1-750DP, significantly less TCoV were detected by IFA in the turkeys in G2-100DPH receiving an extra dose of 100μg DNA mixed with PEI and HA. The results indicated that DNA-prime protein-boost DNA vaccination regimen targeting TCoV S fragment encompassing neutralizing epitopes induced humoral immune response and partially protected turkeys against infectious challenge by TCoV.


title: Human Coronavirus in the 2014 Winter Season as a Cause of Lower Respiratory Tract Infection

text: PURPOSE: During the late autumn to winter season (October to December) in the Republic of Korea, respiratory syncytial virus (RSV) is the most common pathogen causing lower respiratory tract infections (LRTIs). Interestingly, in 2014, human coronavirus (HCoV) caused not only upper respiratory infections but also LRTIs more commonly than in other years. Therefore, we sought to determine the epidemiology, clinical characteristics, outcomes, and severity of illnesses associated with HCoV infections at a single center in Korea. MATERIALS AND METHODS: We retrospectively identified patients with positive HCoV respiratory specimens between October 2014 and December 2014 who were admitted to Severance Children’s Hospital at Yonsei University Medical Center for LRTI. Charts of the patients with HCoV infection were reviewed and compared with RSV infection. RESULTS: During the study period, HCoV was the third most common respiratory virus and accounted for 13.7% of infections. Coinfection was detected in 43.8% of children with HCoV. Interestingly, one patient had both HCoV-OC43 and HCoV-NL63. Mild pneumonia was most common (60.4%) with HCoV, and when combined with RSV, resulted in bronchiolitis. Two patients required care in the intensive care unit. However, compared with that of RSV infection, the disease course HCoV was short. CONCLUSION: Infections caused by HCoVs are common, and can cause LRTIs. During an epidemic season, clinicians should be given special consideration thereto. When combined with other medical conditions, such as neurologic or cardiologic diseases, intensive care unit (ICU) care may be necessary.


docid: uzjtycsz

title: Cloning, expression and immunoassay detection of ferret IFN-γ

text: Ferrets (Mustela putorius furo) develop symptoms upon influenza infection that resemble those of humans, including sneezing, body temperature variation and weight loss. Highly pathogenic strains of influenza A, such as H5N1, have the capacity to cause severe illness or death in ferrets. The use of ferrets as a model of influenza infection is currently limited by a lack of species-specific immunological reagents. Interferon gamma (IFN-γ) plays a key role in the development of innate and adaptive immunity and the regulation of Th1-type immune responses. Here we describe the cloning of the full-length cDNA for ferret IFN-γ. Multiple sequence alignment of the predicted amino acid sequence with those of other species indicates that the predicted ferret protein shares the highest identity with Eurasian badger IFN-γ. We raised two hybridoma clones expressing monoclonal antibodies against recombinant ferret IFN-γ capable of detecting IFN-γ protein derived from mitogen-stimulated ferret PBMCs by immunoblotting, ELISA and ELISPOT assay. Finally, an ELISA utilizing the ferret-specific antibodies detected elevated levels of IFN-γ in serum samples from H3N2 influenza A-infected ferrets.


title: COVID‐19 and medical litigation: More than just the obvious

text: COVID‐19 has massively changed the health landscape around the world. Wide‐ranging changes to healthcare delivery have occurred, especially in hospitals and EDs. Health services have made local decisions about care pathways, in some cases deviating from what would, until recently, have been considered widely accepted care. These changes bring with them new medicolegal risk for clinicians. In Australia, civil liability Acts provide protection for professionals when the criterion of having undertaken ‘competent’ practice that would be ‘widely accepted’ ‘in the circumstances’ is met. There is doubt how courts, and the medical experts who advise them, will evaluate clinical care provided during the pandemic when health services have developed local care pathways and there is no nationally accepted standard.


title: Characterization of transmissible gastroenteritis coronavirus S protein expression products in avirulent S. typhimurium Δcya Δcrp: persistence, stability and immune response in swine

text: Abstract The spike protein from transmissible gastroenteritis virus (TGEV) was expressed in attenuated S. typhimurium Δcya Δcrp Δasd χ3987. Three partially overlapping fragments of TGEV S gene, encoding the amino-terminal, intermediate, and carboxy-terminal end of the protein, as well as the full length gene were inserted into the asd + plasmid pYA292 to generate recombinant plasmids pYATS-1, pYATS-2, pYATS-3, and pYATS-4, respectively, which were transformed into S. typhimurium χ3987. Recombinant S. typhimurium χ3987 (pYATS-1) and χ3987 (pYATS-4) expressing constitutively a 53 kDa amino-terminal fragment of the S protein and the full length protein (144 kDa), respectively, showed high stability. After 50 generations in vitro 60% and 20% of the bacteria transformed with pYATS-1 and pYATS-4, respectively, expressed the S-protein antigen. Since S. typhimurium χ3987 (pYATS-1) showed a better level of expression and stability in vitro, this recombinant strain was selected as a potential bivalent vector to induce both immunity to Salmonella and TGEV in swine. In order to study colonization of swine tissues by S. typhimurium Δcya Δcrp, a gene conferring resistance to rifampicin was cloned into the chromosome of S. typhimurium χ3987, generating χ4509 strain. Both S. typhimurium χ4509 (pYA292) and χ4509 (pYATS-1) colonized the ileum of orally inoculated swine with clearance of bacteria between days 10–20 post-infection. The expression of the amino-terminal fragment of the S protein diminished the ability of S. typhimurium χ4509 (pYATS-1) to colonize deep tissues. The recombinant strain S. typhimurium χ3987 (pYATS-1) induced TGEV specific antibodies in both serum and saliva of orally inoculated swine. This strain, as well as S. typhimurium χ3987 (pYA292), also elicited both systemic and mucosal immunity to Salmonella antigens.


docid: fegkacyz

title: The Thoracic Surgery Social Media Network Experience During the COVID-19 Pandemic

text: [Figure: see text]


title: Tailoring Subunit Vaccine Immunity with Adjuvant Combinations and Delivery Routes Using the Middle East Respiratory Coronavirus (MERS-CoV) Receptor-Binding Domain as an Antigen

text: The development of an effective vaccine is critical for prevention of a Middle East respiratory syndrome coronavirus (MERS-CoV) pandemic. Some studies have indicated the receptor-binding domain (RBD) protein of MERS-CoV spike (S) is a good candidate antigen for a MERS-CoV subunit vaccine. However, highly purified proteins are typically not inherently immunogenic. We hypothesised that humoral and cell-mediated immunity would be improved with a modification of the vaccination regimen. Therefore, the immunogenicity of a novel MERS-CoV RBD-based subunit vaccine was tested in mice using different adjuvant formulations and delivery routes. Different vaccination regimens were compared in BALB/c mice immunized 3 times intramuscularly (i.m.) with a vaccine containing 10 µg of recombinant MERS-CoV RBD in combination with either aluminium hydroxide (alum) alone, alum and polyriboinosinic acid (poly I:C) or alum and cysteine-phosphate-guanine (CpG) oligodeoxynucleotides (ODN). The immune responses of mice vaccinated with RBD, incomplete Freund’s adjuvant (IFA) and CpG ODN by a subcutaneous (s.c.) route were also investigated. We evaluated the induction of RBD-specific humoral immunity (total IgG and neutralizing antibodies) and cellular immunity (ELISpot assay for IFN-γ spot-forming cells and splenocyte cytokine production). Our findings indicated that the combination of alum and CpG ODN optimized the development of RBD-specific humoral and cellular immunity following subunit vaccination. Interestingly, robust RBD-specific antibody and T-cell responses were induced in mice immunized with the rRBD protein in combination with IFA and CpG ODN, but low level of neutralizing antibodies were elicited. Our data suggest that murine immunity following subunit vaccination can be tailored using adjuvant combinations and delivery routes. The vaccination regimen used in this study is promising and could improve the protection offered by the MERS-CoV subunit vaccine by eliciting effective humoral and cellular immune responses.


JAACAP’s Role in Advancing the Science of Pediatric Mental Health and Promoting the Care of Youth and Families During the COVID-19 Pandemic


title: Risk Factors for Death from Influenza A(H1N1)pdm09, State of São Paulo, Brazil, 2009

text: This case-control study aimed to assess the risk factors for death from influenza A(H1N1)pdm09 in patients with laboratory confirmation, who had severe acute respiratory illness-SARI and were hospitalized between June 28(th) and August 29(th) 2009, in the metropolitan regions of São Paulo and Campinas, Brazil. Medical charts of all the 193 patients who died (cases) and the 386 randomly selected patients who recovered (controls) were investigated in 177 hospitals. Household interviews were conducted with those who had survived and the closest relative of those who had died. 73.6% of cases and 38.1% of controls were at risk of developing influenza-related complications. The 18-to-59-year age group (OR = 2.31, 95%CI: 1.31–4.10 (reference up to 18 years of age)), presence of risk conditions for severity of influenza (OR = 1.99, 95%CI: 1.11–3.57, if one or OR = 6.05, 95%CI: 2.76–13.28, if more than one), obesity (OR = 2.73, 95%CI: 1.28–5.83), immunosuppression (OR = 3.43, 95%CI: 1.28–9.19), and search for previous care associated with the hospitalization (OR = 3.35, 95%CI: 1.75–6.40) were risk factors for death. Antiviral treatment performed within 72 hours of the onset of symptoms (OR = 0.17, 95%CI: 0.08–0.37, if within 48hours, and OR = 0.30, 95%CI: 0.11–0.81, if between 48 and 72 hours) was protective against death. The identification of high-risk patients and early treatment are important factors for reducing morbi-mortality from influenza.


Respuesta autores a: A propósito de la carta científica "Sospecha de abdomen agudo como manifestación extrapulmonar de infección COVID-19”


Acute care surgery and post‐operative COVID‐19 care: a comment


title: COVID-19 Infection and Haematological Involvement: a Review of Epidemiology, Pathophysiology and Prognosis of Full Blood Count Findings

text: The unfolding COVID-19 pandemic began in December 2019 in Wuhan, Hubei Province. COVID-19 is a systemic infection affecting several systems including the haematopoietic system. Surveys illustrating the laboratory findings of these patients conclude that lymphocytopenia, neutrophilia and thrombocytopenia are prominent amongst them. Moreover, it has been reported a significant decrease in T lymphocyte subsets and an increase of inflammatory cytokines of hospitalized patients with COVID-19. Generally, thrombocytopenia is commonplace in critically ill patients and usually suggests serious organ malfunction. In view of this, this review investigates the correlation between these abnormalities and the prognosis and disease course. Full blood count is an easy, economic and widely available tool which may help to discriminate between patients with or without severe disease. Last but not least, this review examines potential pathophysiological mechanisms by the novel coronavirus which contribute to these haematological alterations aiding the clinicians to better understand this disease and provide more clinical treatment options.


title: MERS CoV Infection in Two Renal Transplant Recipients: Case Report

text: Middle East Respiratory Syndrome Coronavirus (MERS CoV) infection has recently emerged as a cause of severe potentially fatal pneumonia. The clinical presentation ranges from asymptomatic infection to severe pneumonia and acute renal failure. Data on the clinical presentation in solid organ transplant recipients are lacking. We report two cases of MERS CoV infections in two renal transplant recipients with variable clinical presentations and outcomes. The first patient presented with progressive respiratory symptoms, acute renal failure and died. While the second patient presented with respiratory tract symptoms, remained stable and had an excellent clinical recovery despite recent reception of thymoglobulin induction. This is a rare report of MERS CoV infection in renal transplant recipients. Further data are needed to gain better understanding of the impact of anti‐rejection immunosuppressive therapy on the clinical presentation, severity and outcome of MERS CoV infections in solid organ transplant recipients.


title: Transmission of SARS-CoV-2 via fecal-oral and aerosols–borne routes: Environmental dynamics and implications for wastewater management in underprivileged societies

text: Abstract The advent of novel human coronavirus (SARS-CoV-2) and its potential transmission via fecal-oral and aerosols-borne routes are upcoming challenges to understand the fate of the virus in the environment. In this short communication, we specifically looked at the possibilities of these transmission routes based on the available literature directly related to the SARS-CoV-2 as well as on the closer phylogenetic relatives such as SARS-CoV-1. The available data suggest that, in addition to human-to-human contact, the virus may spread via fecal-oral and aerosols-borne routes. Existing knowledge states that coronaviruses have low stability in the environment due to the natural action of oxidants that disrupt the viral envelope. Previous recommended dosage of chlorination has been found to be not sufficient to inactivate SARS-CoV-2 in places where viral load is high such as hospitals and airports. Although there is no current evidence showing that coronaviruses can be transmitted through contaminated drinking water, there is a growing concern on the impact of the current pandemic wave on underprivileged societies because of their poor wastewater treatment infrastructures, overpopulation, and outbreak management strategies. More research is encouraged to trace the actual fate of SARS-CoV-2 in the environment and to develop/revise the disinfection strategies accordingly.


docid: ywbyflas

title: The evolution of pulmonary pathology in fatal COVID-19 disease: an autopsy study with clinical correlation

text: The pandemia of coronavirus disease 2019 (COVID-19) has caused more than 355,000 confirmed deaths worldwide. However, publications on postmortem findings are scarce. We present the pulmonary findings in four cases of fatal COVID-19 with a spectrum of lung pathology reflecting disease course and duration, invasive therapies, and laboratory features. Early disease is characterized by neutrophilic, exudative capillaritis with microthrombosis and high levels of IL-1beta and IL-6. Later stages are associated with diffuse alveolar damage and ongoing intravascular thrombosis in small to medium-sized pulmonary vessels, occasionally with areas of infarction equivalents, accompanied by laboratory features of disseminated intravascular coagulation. In late stages, organizing pneumonia with extensive intra-alveolar proliferation of fibroblasts and marked metaplasia of alveolar epithelium can be observed. Viral RNA is encountered in the lung, with virus particles in endothelial cells and pneumocytes. In many patients, multi-organ failure with severe liver damage sets in finally, possibly as consequence of an early-onset pro-inflammatory cytokine storm and/or thrombotic microangiopathy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00428-020-02881-x) contains supplementary material, which is available to authorized users.


title: Bioactivities of Phenolics by Focusing on Suppression of Chronic Diseases: A Review

text: Phenolics, which are secondary metabolites of plants, exhibit remarkable bioactivities. In this contribution, we have focused on their protective effect against chronic diseases rather than their antioxidant activities, which have been widely discussed in the literature. A large body of epidemiological studies has proven the bioactivities of phenolics in both standard compounds and natural extracts: namely, anticancer, anti-inflammatory, and antibacterial activities as well as reducing diabetes, cardiovascular disease, and neurodegenerative disease. Phenolics also display anti-analgesic, anti-allergic, and anti-Alzheimer’s properties. Thus, this review provides crucial information for better understanding the bioactivities of phenolics in foods and fills a gap in the existing collective and overall knowledge in the field.


title: COVID-19, a worldwide public health emergency

text: Abstract A new coronavirus outbreak emerged on the 31st of December 2019 in Wuhan, China, causing commotion among the medical community and the rest of the world. This new species of coronavirus has been termed 2019-nCoV and has caused a considerable number of cases of infection and deaths in China and, to a growing degree, beyond China, becoming a worldwide public health emergency. 2019-nCoV has high homology to other pathogenic coronaviruses, such as those originating from bat-related zoonosis (SARS-CoV), which caused approximately 646 deaths in China at the start of the decade. The mortality rate for 2019-nCoV is not as high (approximately 2–3%), but its rapid propagation has resulted in the activation of protocols to stop its spread. This pathogen has the potential to become a pandemic. It is therefore vital to follow the personal care recommendations issued by the World Health Organization.


title: Asthma prevalence in patients with SARS-CoV-2 virus infection detected by RT-PCR not requiring hospitalization

text: INTRODUCTION: The prevalence of asthma in patients hospitalized with SARS-CoV-2 has been studied and varies widely in the different series. However, the prevalence in SARS-infected patients not requiring hospitalization is not known. The objective of this study was to analyze the presence of asthma in a consecutive series of patients who tested positive in the RT-PCR assay for SARS-CoV-2 and did not require hospital admission. METHODS AND RESULTS: A total of 218 patients (58% of those who tested positive) did not require hospitalization; they had a median age of 45 years (IQR 34–57) and 57% were female. Six patients (2.8%) had a previous diagnosis of asthma. Only one patient developed a mild aggravation of asthma symptoms associated with SARS-CoV-2 infection. CONCLUSIONS: Few patients with asthma were infected by SARS-CoV-2, and this infection was not a significant cause of asthma exacerbation.


title: Protein structure analysis of the interactions between SARS-CoV-2 spike protein and the human ACE2 receptor: from conformational changes to novel neutralizing antibodies

text: The recent severe acute respiratory syndrome, known as Coronavirus Disease 2019 (COVID-19) has spread so much rapidly and severely to induce World Health Organization (WHO) to declare a state of emergency over the new coronavirus SARS-CoV-2 pandemic. While several countries have chosen the almost complete lock-down for slowing down SARS-CoV-2 spread, the scientific community is called to respond to the devastating outbreak by identifying new tools for diagnosis and treatment of the dangerous COVID-19. With this aim, we performed an in silico comparative modeling analysis, which allows gaining new insights into the main conformational changes occurring in the SARS-CoV-2 spike protein, at the level of the receptor-binding domain (RBD), along interactions with human cells angiotensin-converting enzyme 2 (ACE2) receptor, that favor human cell invasion. Furthermore, our analysis provides (1) an ideal pipeline to identify already characterized antibodies that might target SARS-CoV-2 spike RBD, aiming to prevent interactions with the human ACE2, and (2) instructions for building new possible neutralizing antibodies, according to chemical/physical space restraints and complementary determining regions (CDR) mutagenesis of the identified existing antibodies. The proposed antibodies show in silico high affinity for SARS-CoV-2 spike RBD and can be used as reference antibodies also for building new high-affinity antibodies against present and future coronaviruses able to invade human cells through interactions of their spike proteins with the human ACE2. More in general, our analysis provides indications for the set-up of the right biological molecular context for investigating spike RBD–ACE2 interactions for the development of new vaccines, diagnostic kits, and other treatments based on the targeting of SARS-CoV-2 spike protein. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00018-020-03580-1) contains supplementary material, which is available to authorized users.


Severe Acute Respiratory Syndrome: Relapse? Hospital Infection?


Doxycycline or minocycline may be a viable treatment option against SARS-CoV-2


title: Universal health insurance coverage for 1.3 billion people: What accounts for China's success?

text: Abstract China successfully achieved universal health insurance coverage in 2011, representing the largest expansion of insurance coverage in human history. While the achievement is widely recognized, it is still largely unexplored why China was able to attain it within a short period. This study aims to fill the gap. Through a systematic political and socio-economic analysis, it identifies seven major drivers for China's success, including (1) the SARS outbreak as a wake-up call, (2) strong public support for government intervention in health care, (3) renewed political commitment from top leaders, (4) heavy government subsidies, (5) fiscal capacity backed by China's economic power, (6) financial and political responsibilities delegated to local governments and (7) programmatic implementation strategy. Three of the factors seem to be unique to China (i.e., the SARS outbreak, the delegation, and the programmatic strategy.) while the other factors are commonly found in other countries’ insurance expansion experiences. This study also discusses challenges and recommendations for China's health financing, such as reducing financial risk as an immediate task, equalizing benefit across insurance programs as a long-term goal, improving quality by tying provider payment to performance, and controlling costs through coordinated reform initiatives. Finally, it draws lessons for other developing countries.


title: Stool cultures for nosocomial diarrhea: money down the drain?

text: The authors report on a modified “3-day rule” for limiting low-yield stool cultures obtained from hospitalized patients with nosocomial diarrhea. The modified rule addresses the two known weaknesses of the strict 3-day rule: immune-suppressed patients and nosocomial outbreaks of bacterial gastroenteritis. In addition, the modified rule addresses a third group of previously unidentified patients at risk of non-Clostridium difficile bacterial nosocomial diarrhea: patients older than 65 with comorbidity. The modified rule would decrease the number of stool cultures by half without missing any clinically significant cases. The authors recommend the modified 3-day rule as a way to “greatly reduce the laboratory burden of stool culture, yet provide rapid diagnosis for patients at increased risk of nosocomial bacterial gastroenteritis.”


docid: pbhdzshl

title: Coronavirus glycoprotein E1, a new type of viral glycoprotein

text: Abstract The carbohydrate contents of coronavirus glycoproteins E1 and E2 have been analyzed. E2 has complex and mannose-rich-type oligosaccharide side-chains, which are attached by N-glycosidic linkages to the polypeptide. Glycosylation of E2 is initiated at the co-translational level, and it is inhibited by tunicamycin, 2-deoxy-glucose, and 2-deoxy-2-fluoro-glucose. Thus, E2 belongs to a glycoprotein type found in many other enveloped viruses. E1, in contrast, represents a different class of glycoprotein. The following observations indicate that its carbohydrate side-chains have 0-glycosidic linkage. (1) The constituent sugars of E1 are N-acetylglucosamine, N-acetylgalactosamine, galactose, and neuraminic acid; mannose and fucose are absent. (2) The side-chains can be removed by β-elimination. (3) Glycosylation of E1 is not sensitive to the compounds interfering with N-glycosylation. E1 is the first viral glycoprotein analyzed that contains only 0-glycosidic linkages. Coronaviruses are therefore a suitable model system to study biosynthesis and processing of this type of glycoprotein.


title: Chapter 7 Global Supply of Virus-Safe Drinking Water

text: Publisher Summary This chapter illustrates the recommendations and guidelines of the World Health Organization (WHO) concerning water, sanitation, and health. The recommendations and guidelines are evaluated in the light of disease caused by human pathogenic viruses. The guidelines outline a preventive management framework for safe drinking water. The framework includes health-based targets to assist national authorities who are normally responsible to set the targets for the protection of public health from risks by exposure to drinking water. Assessing the adequacy of systems, defining and monitoring control measures, and establishing management plans are the three components of the so-called water safety plans. Achievement of health-based targets may be verified by independent surveillance to assess the safety of the drinking water through additional verification or audit-based approaches. This framework for safe drinking water can be adapted according to environmental, social, economic, and cultural circumstances of drinking water provision on the national, regional, and local level. The chapter concludes that viruses could be considered as biocolloids with specific properties such as size, shape, structure, charge, composition, and genome. These viral characteristics determine their behavior in the environment, resistance to natural inactivation and treatment, and disinfection processes. For each (re-)emerging virus these properties may be known or could be assessed predicting the effectiveness of possible intervention measures for prevention of waterborne disease.


title: Psychologist Self-Care During the Pandemic: Now More Than Ever

text: The COVID-19 crisis has transformed the lives and practices of psychologists and has highlighted the need for time-efficient self-care. The anxiety among psychologists (and humans) is practically universal—nearly every psychologist worldwide is experiencing some negative impact on their mental health. We offer nine research-supported, practitioner-friendly methods of self-care to manage anxiety and maintain optimal performance during these extraordinary times.


Screening of Donor and Recipient Prior to Solid Organ Transplantation


title: ANGIOTENSIN CONVERTING ENZYME 2 IN THE KIDNEY

text: 1.. Angiotensin converting enzyme 2 (ACE2) is an important homeostatic component of the renin angiotensin system (RAS). ACE2 both degrades the vasoconstrictor, angiotensin II and generates the potent vasodilator peptide, angiotensin 1–7. These actions counterbalance those of ACE. 2.. ACE2 is highly expressed in the healthy kidney, particularly in the proximal tubules, where it colocalizes with ACE and angiotensin receptors. 3.. Kidney disease and subtotal nephrectomy is associated with a reduction in renal ACE2 expression, possibly facilitating the damaging effects of angiotensin II in the failing kidney. Acquired or genetic ACE2 deficiency also appears to exacerbate renal damage and albuminuria in experimental models, supporting this hypothesis. 4.. ACE2 also has an important role in blood pressure control. Many models of hypertension are associated with reduced ACE2 expression. Although ACE2 KO animals are normotensive, in states associated with activation of the RAS, ACE2 overexpression improves blood pressure control and reduces angiotensin responsiveness.


Trials of BCG vaccine will test for covid-19 protection


docid: njkhwkwk

title: Potential spreading risks and disinfection challenges of medical wastewater by the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) viral RNA in septic tanks of Fangcang Hospital

text: Abstract The outbreak of coronavirus infectious disease-2019 (COVID-19) pneumonia raises the concerns of effective deactivation of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in medical wastewater by disinfectants. In this study, we evaluated the presence of SARS-CoV-2 viral RNA in septic tanks of Wuchang Cabin Hospital and found a striking high level of (0.5–18.7) × 103 copies/L after disinfection with sodium hypochlorite. Embedded viruses in stool particles might be released in septic tanks, behaving as a secondary source of SARS-CoV-2 and potentially contributing to its spread through drainage pipelines. Current recommended disinfection strategy (free chlorine ≥0.5 mg/L after at least 30 min suggested by World Health Organization; free chlorine above 6.5 mg/L after 1.5-h contact by China Centers for Disease Control and Prevention) needs to be reevaluated to completely remove SARS-CoV-2 viral RNA in non-centralized disinfection system and effectively deactivate SARS-CoV-2. The effluents showed negative results for SARS-CoV-2 viral RNA when overdosed with sodium hypochlorite but had high a level of disinfection by-product residuals, possessing significant ecological risks.


title: The relationship between obsessive compulsive beliefs and symptoms, anxiety and disgust sensitivity, and Swine Flu fears

text: Abstract Using the 2009 Swine Flu outbreak as a contemporary example of pandemic fears, this study examined the relationship between various symptoms related to anxiety sensitivity and Swine Flu fears. It was hypothesized that both obsessive–compulsive (OC) beliefs and OC symptoms would significantly predict Swine Flu fears. It was also hypothesized that symptoms of anxiety, including measures of anxiety sensitivity and disgust sensitivity would significantly mediate the relationship between both OC beliefs and OC symptoms and Swine Flu fears. A total of 393 undergraduate students completed measures of Swine Flu fears, anxiety sensitivity, OC beliefs and symptoms, and disgust sensitivity. It was found that both OC beliefs and OC symptoms significantly predicted Swine Flu fears. While disgust sensitivity significantly mediated the relationship between both OC beliefs and OC symptoms and Swine Flu fears using the Sobel test, anxiety sensitivity was a significant mediator only for OC symptoms. Additionally, path modeling showed that anxiety sensitivity mediated the relationship between OC symptoms and Swine Flu fears best. The results of this study may be useful for treating individuals suffering from anxiety in light of future pandemics, as well as continuing to research the role of anxiety symptoms in predicting pandemic fears.


title: Modeling Nigerian Covid-19 cases: A comparative analysis of models and estimators

text: COVID-19 remains a major pandemic currently threatening all the countries of the world. In Nigeria, there were 1, 932 COVID-19 confirmed cases, 319 discharged cases and 58 deaths as of 30th April 2020. This paper, therefore, subjected the daily cumulative reported COVID-19 cases of these three variables to nine (9) curve estimation statistical models in simple, quadratic, cubic, and quartic forms. It further identified the best of the thirty-six (36) models and used the same for prediction and forecasting purposes. The data collected by the Nigeria Centre for Disease Control for sixty-four (64) days, two (2) months and three (3), were daily monitored and eventually analyzed. We identified the best models to be Quartic Linear Regression Model with an autocorrelated error of order 1 (AR(1)); and found the Ordinary Least Squares, Cochrane Orcutt, Hildreth–Lu, and Prais-Winsten and Least Absolute Deviation (LAD) estimators useful to estimate the models’ parameters. Consequently, we recommended the daily cumulative forecast values of the LAD estimator for May and June 2020 with a 99% confidence level. The forecast values are alarming, and so, the Nigerian Government needs to hastily review her activities and interventions towards COVID-19 to provide some tactical and robust structures and measures to avert these challenges.


title: Chapter Seven Vaccine Adjuvant Nanotechnologies

text: Abstract The increasing sophistication of vaccine adjuvant design has been driven by improved understanding of the importance of nanoscale features of adjuvants to their immunological function. Newly available advanced nanomanufacturing techniques now allow very precise control of adjuvant particle size, shape, texture, and surface chemistry. Novel adjuvant concepts include self-assembling particles and targeted immune delivery. These individual concepts can be combined to create a single integrated vaccine nanoparticle-combining antigen, adjuvants, and DC-targeting elements. In the process, the concept of an adjuvant has broadened to include not only immune-stimulatory substances but also any design features that enhance the immune response against the relevant vaccine antigen. The modern definition of an adjuvant includes not only classical immune stimulators but also any aspects of particle size, shape, and surface chemistry that enhance vaccine immunogenicity. It even includes purely physical processes such as texturing of particle surfaces to maximize immunogenicity. Looking forward, adjuvants will increasingly be seen not as separate add-on items but as wholly integrated elements of a complete vaccine delivery package. Hence, vaccine systems will increasingly approach the complexity and sophistication of pathogens themselves, incorporating highly specific particle properties, contents, and behaviors, all designed to maximize immune system recognition and drive the immune response in the specific direction that affords maximal protection.


title: Décision kinésithérapique: Michelle T. 73 ans. Réhabilitation respiratoire, en centre de soins de suite et de réadaptation, nécessaire malgré une bonne santé avant la maladie Covid-19

text: Abstract This clinical study covers the care of a Covid-positive patient after she was admitted in a pulmonary rehabilitation centre. The clinical pattern is mainly marked by dyspnea, but also by the psychological trauma caused by impairment and hospitalization in intensive care. The patient's previous quality of life, the lack of comorbidity and a tailored multidisciplinary care will be decisive in her rehabilitation progress.


title: The Relationship between the Migrant Population’s Migration Network and the Risk of COVID-19 Transmission in China—Empirical Analysis and Prediction in Prefecture-Level Cities

text: The outbreak of COVID-19 in China has attracted wide attention from all over the world. The impact of COVID-19 has been significant, raising concerns regarding public health risks in China and worldwide. Migration may be the primary reason for the long-distance transmission of the disease. In this study, the following analyses were performed. (1) Using the data from the China migrant population survey in 2017 (Sample size = 432,907), a matrix of the residence–birthplace (R-B matrix) of migrant populations is constructed. The matrix was used to analyze the confirmed cases of COVID-19 at Prefecture-level Cities from February 1–15, 2020 after the outbreak in Wuhan, by calculating the probability of influx or outflow migration. We obtain a satisfactory regression analysis result (R(2) = 0.826–0.887, N = 330). (2) We use this R-B matrix to simulate an outbreak scenario in 22 immigrant cities in China, and propose risk prevention measures after the outbreak. If similar scenarios occur in the cities of Wenzhou, Guangzhou, Dongguan, or Shenzhen, the disease transmission will be wider. (3) We also use a matrix to determine that cities in Henan province, Anhui province, and Municipalities (such as Beijing, Shanghai, Guangzhou, Shenzhen, Chongqing) in China will have a high risk level of disease carriers after a similar emerging epidemic outbreak scenario due to a high influx or outflow of migrant populations.


title: Use of Sentinel Laboratories by Clinicians to Evaluate Potential Bioterrorism and Emerging Infections

text: With the persistent threat of emerging infectious diseases and bioterrorism, it has become increasingly important that clinicians be able to identify the diseases that might signal the occurrence of these unusual events. Essential to a thoughtful diagnostic approach is understanding when to initiate a public health investigation and how to appropriately use commonly performed microbiology procedures in the sentinel laboratory to evaluate potential pathogens. Although diagnostic test development is evolving rapidly, recognizing many of these pathogens continues to challenge the capabilities of most sentinel laboratories. Therefore, effective, ongoing communication and education among clinicians, infection control personnel, sentinel laboratorians, public health authorities, and Laboratory Response Network reference laboratorians is the key to preparedness.


title: Evolutionary and Phylogenetic Analysis of the Hepaciviruses and Pegiviruses

text: The known genetic diversity of the hepaciviruses and pegiviruses has increased greatly in recent years through the discovery of viruses related to hepatitis C virus and human pegivirus in bats, bovines, equines, primates, and rodents. Analysis of these new species is important for research into animal models of hepatitis C virus infection and into the zoonotic origins of human viruses. Here, we provide the first systematic phylogenetic and evolutionary analysis of these two genera at the whole-genome level. Phylogenies confirmed that hepatitis C virus is most closely related to viruses from horses whereas human pegiviruses clustered with viruses from African primates. Within each genus, several well-supported lineages were identified and viral diversity was structured by both host species and location of sampling. Recombination analyses provided evidence of interspecific recombination in hepaciviruses, but none in the pegiviruses. Putative mosaic genome structures were identified in NS5B gene region and were supported by multiple tests. The identification of interspecific recombination in the hepaciviruses represents an important evolutionary event that could be clarified by future sampling of novel viruses. We also identified parallel amino acid changes shared by distantly related lineages that infect similar types of host. Notable parallel changes were clustered in the NS3 and NS4B genes and provide a useful starting point for experimental studies of the evolution of Hepacivirus host–virus interactions.


title: On-chip analysis of respiratory viruses from nasopharyngeal samples

text: Point-of-care (PoC) testing followed by personalized efficient therapy of infectious diseases may result in a considerable reduction of associated health care costs. Lab-on-a-chip (LoC) systems represent a potentially high efficient class of PoC tools. Here, we present a LoC system for automated pathogen analysis of respiratory viruses from nasopharyngeal specimens. The device prepares total nucleic acids from extracted swab samples using magnetic silica beads. After reverse transcription the co-purified viral RNA is amplified in accordance with the QIAplex multiplex PCR technology. Hybridized to corresponding QIAGEN LiquiChip beads and labelled with streptavidin R-phycoerythrin, the amplified target sequences are finally detected using a QIAGEN LiquiChip200 workstation. All chemicals needed are either stored freeze-dried on the disposable chip or are provided in liquid form in a reagent cartridge for up to 24 runs. Magnetic stir bars for mixing as well as turning valves with metering structures are integrated into the injection-moulded disposable chip. The core of the controlling instrument is a rotating heating bar construction providing fixed temperatures for fast cycling. PCR times of about half an hour (for 30 cycles) could be achieved for 120 μl reactions, making this system the fastest currently available high-volume PCR chip. The functionality of the system was shown by comparing automatically processed nasopharyngeal samples to ones processed manually according to the QIAGEN “ResPlex™ II Panel v2.0” respiratory virus detection kit. A prototype of the present instrument revealed slightly weaker signal intensities with a similar sensitivity in comparison to the commercially available kit and automated nucleic acid preparation devices, even without protocol optimization.


title: Prevalence of serum IgG antibodies against SARS-CoV-2 among clinic staff

text: The SARS-CoV-2 pandemic threatens health care providers and society. For planning of treatment capacities, it is of major importance to obtain reliable information on infection and fatality rates of the novel coronavirus. A German community study, the so-called Heinsberg study, found a 5-fold higher infection rate (and thus a remarkably lower fatality rate) than the officially reported cases suggest. We were interested to examine the SARS-CoV-2-IgG antibody status among clinic staff of a large neurological center in Northern Germany. Blood samples and questionnaires (demographic data, medical history) were collected pseudonymously. In total, 406 out of 525 (77.3%) of our employees participated in the study. The infection rate among the staff was as high as 2.7%. Including drop-outs (missing questionnaire but test result available), the infection rate was even higher (2.9%). Only 36% of the positively tested employees did suffer from flu-like symptoms in 2020. None of the nurses–having closest and longest contact to patients—were found to be positive. Despite the fact that the infection rate among clinic staff may not be directly compared to the situation in the surrounding county (due to different testing procedures), one might hypothesize that the infection rate could be more than 30-fold higher than the number of officially reported cases for the county of Hameln-Pyrmont. The high rate of IgG-positive, asymptomatic healthcare workers might help to overcome fears in daily work.


title: The immunology of COVID-19: is immune modulation an option for treatment?

text: In December, 2019, an outbreak of COVID-19 emerged in Wuhan, China and quickly spread globally. As of May 7, 2020, there were 3 672 238 confirmed infections and 254 045 deaths attributed to COVID-19. Evidence has shown that there are asymptomatic carriers of COVID-19 who can transmit the disease to others. The virus incubation time shows a wide range (0–24 days) and the virus displays a high infectivity. It is therefore urgent to develop an effective therapy to treat patients with COVID-19 and to control the spread of the causative agent, severe respiratory syndrome coronavirus 2. Repurposing of approved drugs is widely adopted to fight newly emerged diseases such as COVID-19, as these drugs have known pharmacokinetic and safety profiles. As pathological examination has confirmed the involvement of immune hyperactivation and acute respiratory distress syndrome in fatal cases of COVID-19, several disease-modifying anti-rheumatic drugs (DMARDS), such as hydroxychloroquine and tocilizumab, have been proposed as potential therapies for the treatment of COVID-19. In this Review, we discuss the immunological aspects of COVID-19 and the potential implication of DMARDs in treating this disease.


title: Zero COVID‐19 infection in inflammatory bowel disease patients: Findings from population‐based inflammatory bowel disease registries in Hong Kong and Taiwan

text: BACKGROUND AND AIM: It is unsure whether inflammatory bowel disease (IBD) is a risk factor for novel coronavirus infection (COVID‐19). METHODS: IBD patients were identified from population‐based databases in Hong Kong and Taiwan from January 21, 2020, until April 15, 2020. RESULTS: Total 2954 and 2554 IBD patients were identified in Hong Kong and Taiwan, respectively. None had COVID‐19. Pooled analysis showed that 65.3%, 39.1%, 4.3%, and 12.8% IBD patients in Hong Kong and 75.8 %, 51.4 %, 26.1%, and 52.3 % in Taiwan were on 5‐aminosalicylates, immunomodulators, corticosteroids, and biologics, respectively. CONCLUSION: There were no reported cases of COVID‐19 infection amongst IBD patients in Hong Kong and Taiwan. IBD patients should continue their usual medications during the COVID‐19 pandemic.


title: Insights into N-calls of mitochondrial DNA sequencing using MitoChip v2.0

text: BACKGROUND: Developments in DNA resequencing microarrays include mitochondrial DNA (mtDNA) sequencing and mutation detection. Failure by the microarray to identify a base, compared to the reference sequence, is designated an 'N-call.' This study re-examined the N-call distribution of mtDNA samples sequenced by the Affymetrix MitoChip v.2.0, based on the hypothesis that N-calls may represent insertions or deletions (indels) in mtDNA. FINDINGS: We analysed 16 patient mtDNA samples using MitoChip. N-calls by the proprietary GSEQ software were significantly reduced when either of the freeware on-line algorithms ResqMi or sPROFILER was utilized. With sPROFILER, this decrease in N-calls had no effect on the homoplasmic or heteroplasmic mutation levels compared to GSEQ software, but ResqMi produced a significant change in mutation load, as well as producing longer N-cell stretches. For these reasons, further analysis using ResqMi was not attempted. Conventional DNA sequencing of the longer N-calls stretches from sPROFILER revealed 7 insertions and 12 point mutations. Moreover, analysis of single-base N-calls of one mtDNA sample found 3 other point mutations. CONCLUSIONS: Our study is the first to analyse N-calls produced from GSEQ software for the MitoChipv2.0. By narrowing the focus to longer stretches of N-calls revealed by sPROFILER, conventional sequencing was able to identify unique insertions and point mutations. Shorter N-calls also harboured point mutations, but the absence of deletions among N-calls suggests that probe confirmation affects binding and thus N-calling. This study supports the contention that the GSEQ is more capable of assigning bases when used in conjunction with sPROFILER.


Initial resolution of the COVID-19 pandemic in China—can the virus return?