title: Mapping infectious disease hospital surge threats to lessons learnt in Singapore: a systems analysis and development of a framework to inform how to DECIDE on planning and response strategies

text: BACKGROUND: Hospital usage and service demand during an Infectious Disease (ID) outbreak can tax the health system in different ways. Herein we conceptualize hospital surge elements, and lessons learnt from such events, to help build appropriately matched responses to future ID surge threats. METHODS: We used the Interpretive Descriptive qualitative approach. Interviews (n = 35) were conducted with governance and public health specialists; hospital based staff; and General Practitioners. Key policy literature in tandem with the interview data were used to iteratively generate a Hospital ID Surge framework. We anchored our narrative account within this framework, which is used to structure our analysis. RESULTS: A spectrum of surge threats from combinations of capacity (for crowding) and capability (for treatment complexity) demands were identified. Starting with the Pyramid scenario, or an influx of high screening rates flooding Emergency Departments, alongside fewer and manageable admissions; the Reverse-Pyramid occurs when few cases are screened and admitted but those that are, are complex; during a ‘Black’ scenario, the system is overburdened by both crowding and complexity. The Singapore hospital system is highly adapted to crowding, functioning remarkably well at constant near-full capacity in Peacetime and resilient to Endemic surges. We catalogue 26 strategies from lessons learnt relating to staffing, space, supplies and systems, crystalizing institutional memory. The DECIDE model advocates linking these strategies to types of surge threats and offers a step-by-step guide for coordinating outbreak planning and response. CONCLUSIONS: Lack of a shared definition and decision making of surge threats had rendered the procedures somewhat duplicative. This burden was paradoxically exacerbated by a health system that highly prizes planning and forward thinking, but worked largely in silo until an ID crisis hit. Many such lessons can be put into play to further strengthen our current hospital governance and adapted to more diverse settings.


title: Human Infection with Influenza Virus A(H10N8) from Live Poultry Markets, China, 2014

text: Human infection with avian influenza virus A(H10N8) was initially reported in China in December 2013. We characterized H10N8 strains from a human patient and from poultry in live markets that infected persons had visited. Results of genome sequencing and virus characterization suggest that the virus strains that infected humans originated from these markets.


title: Coronavirus infection in mink (Mustela vision). Serological evidence of infection with a coronavirus related to transmissible gastroenteritis virus and porcine epidemic diarrhea virus

text: Abstract Antibodies to a transmissible gastroenteritis virus (TGEV)-related coronavirus have been demonstrated in mink sera by indirect immunofluorescence, peroxidase-linked antibody assays and immunoblotting. This is the first serological evidence of a specific coronavirus infection in mink. The putative mink coronavirus (MCV) seems to be widespread in the Danish mink population with a prevalence approaching 100%. Analysis by immunoblotting has shown that MCV is closely related to TGEV by the spike (S), matrix (M) and nucleoprotein (N) polypeptides. Furthermore, antibodies to MCV also cross-reacted with N and M polypeptides of porcine epidemic diarrhea virus (PEDV). Thus MCV may occupy an intermediate position between the TGEV group of coronaviruses and PEDV. The possibility that MCV may be associated with syndromes of acute enteritis in preweaning mink is discussed.


title: Lessons learned from the anti-SARS quarantine experience in a hospital-based fever screening station in Taiwan

text: BACKGROUND: Severe acute respiratory syndrome (SARS) was the first major novel infectious disease to hit the international community in the 21st century. While SARS was sweeping over almost 30 countries, most hospitals in Taiwan instituted mandatory quarantine measures, one of the most effective public health strategies for preventing disease transmission. We explored the anti-SARS quarantine experience of patients in a hospital-based fever screening station. METHODS: We conducted a phenomenologic, qualitative study using semistructured telephone interviews during the SARS outbreak in Taiwan. Seventeen patients with fever who were quarantined in the fever screening station of a hospital emergency department for at least 2 hours were recruited into this study. RESULTS: Data analysis using Collaizi's 9 steps revealed 2 categories—external burden and internal struggle—and 6 themes regarding patients' quarantine experience. External burden included 3 themes: (1) bearing the uncomfortable surroundings, (2) facing discrimination, and (3) lacking in-person family support. Internal struggle consisted of 3 themes: (1) struggle with being quarantined, (2) struggle with emotional turmoil, and (3) struggle with possible SARS diagnosis. CONCLUSION: These results will contribute to sensitizing health care professionals to empathize with quarantined persons while providing quality quarantine care and other infection control measures.


Consequences of COVID-19 on international medical graduates and students applying to residencies in the United States


title: Replication of sialodacryoadenitis virus in mouse L-2 cells

text: Sialodacryoadenitis (SDA) is a naturally-occurring infection of the laboratory rat raused by the coronavirus, sialodacryoadenitis virus (SDAV). The study of SDAV has been limited because there is no widely available continuous cell line for the propagation of high titers of the virus. The purpose of this study, therefore, was to compare the ability of SDAV to replicate in the permanent cell lines, LBC, of rat origin, and the mouse cell lines. L-929 and L-2. Following 2 to 6 repeated passages of SDAV in LBC cells, the virus could be readily propagated in LBC and L-2 cells, but not in L-929 cells. Similarly, SDAV adapted to replicate directly in L-2 cells could be readily propagated in LBC, but not L-929 cells. In LBC and L-2 cells, cytopathic effect (CPE), viral antigen, viral particles, and virus infectivity could be demonstrated. Titers of up to 10(8.0) infectious viral particles/0.25 ml of culture fluid were obtained at 48 hours in L-2 cells. Titers in LBC cells were one to two logs lower. When susceptible rats were inoculated with eighth passage L-2 cell-adapted virus, they developed typical lesions of SDA. Virus could be recovered from infected tissues and propagated in L-2 cells on first passage. The ability to propagate SDAV to high titers in the widely available L-2 cell line should promote the study of this virus and facilitate its comparison with other murine coronaviruses.


title: The Highest Cited Papers in Brucellosis: Identification Using Two Databases and Review of the Papers' Major Findings

text: Citation classics represent the highest impact work in a given field. We aim to identify and analyze the most frequently cited papers on brucellosis. We used the databases Scopus and Web of Science to determine the most frequently cited papers. The most cited fifty papers in each database were identified. We then ranked the papers according to the highest citation count recorded from any of the two databases. The most frequently cited paper received 964 citations and was by DelVecchio VG et al. reporting the complete genomic sequencing of Brucella melitensis. The papers were published in 30 journals led by the “Infection and Immunity” journal and the “Veterinary Microbiology” journal (each had 7 papers). Citation classics in brucellosis were all in English except one in French and were mostly of basic science type. In addition, we noticed that 12 articles that were identified among the highest fifty articles in one database were missed by the other database and vice versa. Therefore, we suggest that searching in more than one database would detect additional citation classics.


title: Acquisition of multidrug-resistant Enterobacterales during international travel: a systematic review of clinical and microbiological characteristics and meta-analyses of risk factors

text: BACKGROUND: International tourism increased from 25 million tourist arrivals in 1950 to over 1.3 billion in 2017. These travelers can be exposed to (multi) resistant microorganisms, may become colonized, and bring them back home. This systematic review aims to identify the carriage rates of multidrug-resistant Enterobacterales (MDR-E) among returning travelers, to identify microbiological methods used, and to identify the leading risk factors for acquiring MDR-E during international travel. METHODS: Articles related to our research question were identified through a literature search in multiple databases (until June 18, 2019) - Embase, Medline Ovid, Cochrane, Scopus, Cinahl, Web of Science, and Google Scholar. RESULTS: Out of 3211 potentially relevant articles, we included 22 studies in the systematic review, and 12 studies in 7 random-effects meta-analyses. Highest carriage rates of MDR-E were observed after travel to Southern Asia (median 71%), followed by travel to Northern Africa (median 42%). Carbapenemase-producing Enterobacterales (CPE) were identified in 5 out of 22 studies, from a few patients. However, in only eight out of 22 studies (36.4%) the initial laboratory method targeted detection of the presence of CPE in the original samples. The risk factor with the highest pooled odds ratio (OR) for MDR-E was travel to Southern Asia (pooled OR = 14.16, 95% confidence interval [CI] = 5.50 to 36.45), followed by antibiotic use during travel (pooled OR = 2.78, 95% CI = 1.76 to 4.39). CONCLUSIONS: Risk of acquiring MDR-E while travelling increases depending on travel destination and if antibiotics are used during travel. This information is useful for the development of guidelines for healthcare facilities with low MDR-E prevalence rates to prevent admission of carriers without appropriate measures. The impact of such guidelines should be assessed.


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title: A novel plan to deal with SARS‐CoV‐2 and COVID‐19 disease

text: As we write this report in April 2020, more than two million people worldwide have been infected with the SARS-CoV-2 coronavirus and more than 150,000 have died of COVID-19 disease. International economies have been brought to a standstill, and social isolation based on palpable fear of death remains the order of the day. Appropriately or not, the United States government is talking about resuming work activities and social interaction to boost economic recovery. While this makes financial sense, from a medical perspective our population will be left defenseless in the absence of a viable treatment strategy for SARS-CoV-2. Herein we present a plan to deal with this looming threat. This article is protected by copyright. All rights reserved.


title: Amplified ozone pollution in cities during the COVID-19 lockdown

text: Abstract The effect of lockdown due to coronavirus disease (COVID-19) pandemic on air pollution in four Southern European cities (Nice, Rome, Valencia and Turin) and Wuhan (China) was quantified, with a focus on ozone (O3). Compared to the same period in 2017–2019, the daily O3 mean concentrations increased at urban stations by 24% in Nice, 14% in Rome, 27% in Turin, 2.4% in Valencia and 36% in Wuhan during the lockdown in 2020. This increase in O3 concentrations is mainly explained by an unprecedented reduction in NOx emissions leading to a lower O3 titration by NO. Strong reductions in NO2 mean concentrations were observed in all European cities, ~53% at urban stations, comparable to Wuhan (57%), and ~65% at traffic stations. NO declined even further, ~63% at urban stations and ~78% at traffic stations in Europe. Reductions in PM2.5 and PM10 at urban stations were overall much smaller both in magnitude and relative change in Europe (~8%) than in Wuhan (~42%). The PM reductions due to limiting transportation and fuel combustion in institutional and commercial buildings were partly offset by increases of PM emissions from the activities at home in some of the cities. The NOx concentrations during the lockdown were on average 49% lower than those at weekends of the previous years in all cities. The lockdown effect on O3 production was ~10% higher than the weekend effect in Southern Europe and 38% higher in Wuhan, while for PM the lockdown had the same effect as weekends in Southern Europe (~6% of difference). This study highlights the challenge of reducing the formation of secondary pollutants such as O3 even with strict measures to control primary pollutant emissions. These results are relevant for designing abatement policies of urban pollution.


title: Hepatitis E virus ORF1 encoded macro domain protein interacts with light chain subunit of human ferritin and inhibits its secretion

text: Hepatitis E Virus (HEV) is the major causative agent of acute hepatitis in developing countries. Its genome has three open reading frames (ORFs)—called as ORF1, ORF2, and ORF3. ORF1 encodes nonstructural polyprotein having multiple domains, namely: Methyltransferase, Y domain, Protease, Macro domain, Helicase, and RNA-dependent RNA polymerase. In the present study, we show that HEV-macro domain specifically interacts with light chain subunit of human ferritin (FTL). In cultured hepatoma cells, HEV-macro domain reduces secretion of ferritin without causing any change in the expression levels of FTL. This inhibitory effect was further enhanced upon Brefeldin-A treatment. The levels of transferrin Receptor 1 or ferroportin, two important proteins in iron metabolism, remained unchanged in HEV-macro domain expressing cells. Similarly, there were no alterations in the levels of cellular labile iron pool and reactive oxygen species, indicating that HEV-macro domain does not influence cellular iron homeostasis/metabolism. As ferritin is an acute-phase protein, secreted in higher level in infected persons and HEV-macro domain has the property of reducing synthesis of inflammatory cytokines, we propose that by directly binding to FTL, macro domain prevents ferritin from entering into circulation and helps in further attenuation of the host immune response. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11010-016-2715-0) contains supplementary material, which is available to authorized users.


title: Biochemical characterisation of the recombinant peroxiredoxin (FhePrx) of the liver fluke, Fasciola hepatica

text: Abstract The parasitic helminth Fasciola hepatica secretes a 2-Cys peroxiredoxin (Prx) that may play important functions in host–parasite interaction. Recombinant peroxiredoxin (FhePrx) prevented metal-catalyzed oxidative nicking of plasmid DNA and detoxified hydrogen peroxide when coupled with Escherichia coli thioredoxin and thioredoxin reductase (k cat/K m =5.2×105 M−1 s−1). Enzyme kinetic analysis revealed that the catalytic efficiency of FhePrx is similar to other 2-Cys peroxiredoxins; the enzyme displayed saturable enzyme Michaelis–Menten type kinetics with hydrogen peroxide, cumene hydroperoxide and t-butyl hydroperoxide, and is sensitive to concentrations of hydrogen peroxide above 0.5mM. Like the 2-Cys peroxiredoxins from a related helminth, Schistosoma mansoni, steady-state kinetics indicate that FhePrx exhibits a saturable, single displacement-like reaction mechanism rather than non-saturable double displacement (ping–pong) enzyme substitution mechanism common to other peroxiredoxins. However, unlike the schistosome Prxs, FhePrx could not utilise reducing equivalents supplied by glutathione or glutathione reductase.


title: Evaluation of the commercially available LightMix® Modular E-gene kit using clinical and proficiency testing specimens for SARS-CoV-2 detection

text: BACKGROUND: Rapid and sensitive diagnostic assays for SARS-CoV-2 detection are required for prompt patient management and infection control. The analytical and clinical performances of LightMix® Modular SARS and Wuhan CoV E-gene kit, a widely used commercial assay for SARS-CoV-2 detection, have not been well studied. OBJECTIVE: To evaluate the performance characteristics of the LightMix® E-gene kit in comparison with well-validated in-house developed tests. STUDY DESIGN: Serial dilutions of SARS-CoV-2 culture isolate extracts were used for analytical sensitivity evaluation. A total of 289 clinical specimens from 186 patients with suspected COVID-19 and 8 proficiency testing (PT) samples were used to evaluate the diagnostic performance of the LightMix® E-gene kit against in-house developed COVID-19-RdRp/Hel and COVID-19-N RT-PCR assays. RESULTS: The LightMix® E-gene kit had a limit-of-detection of 1.8 × 10(-1) TCID(50)/mL, which was one log(10) lower than those of the two in-house RT-PCR assays. The LightMix® E-gene kit (149/289 [51.6%]) had higher diagnostic sensitivity than the in-house assays (144/289 [49.8%] for RdRp/Hel and 146/289 [50.5%] for N). All three assays gave correct results for all the PT samples. Cycle threshold (Cp) values of the LightMix® E-gene kit and in-house assays showed excellent correlation. Reproducibility of the Cp values was satisfactory with intra- and inter-assay coefficient of variation values <5%. Importantly, the LightMix® E-gene kit, when used as a stand-alone assay, was equally sensitive as testing algorithms using multiple SARS-CoV-2 assays. CONCLUSIONS: The LightMix® E-gene kit is a rapid and sensitive assay for SARS-CoV-2 diagnostics, with fewer verification requirements compared to laboratory-developed tests


title: Nationwide surveillance of 18 respiratory viruses in patients with influenza‐like illnesses: A pilot feasibility study in the French Sentinel Network

text: The aim of the present study was to test the feasibility of integrating the diagnosis of 18 respiratory viruses into clinical surveillance of influenza‐like illness using a PCR‐DNA microarray detection assay. The study took place in the French Sentinel Network, a nationwide surveillance network of General Practitioners (GPs) representative of French GPs in terms age, location, and type of practice (urban/rural). Three virological laboratories also participated in the study. The study was planned for 5 weeks from January 25, 2010 to February 27, 2010. A subset of 150 Sentinel GPs, located in mainland France, was enrolled to collect clinical data and nasopharyngeal samples from every first patient of the week having a medical visit for influenza‐like illness defined as a sudden fever of 39°C or more with respiratory symptoms and myalgia. Sixty‐three GPs (42%) collected 103 samples while 87 GPs (58%) did not. GPs did not differ with respect to their age, gender, urban/rural distribution, or years of inscription in the Sentinel Network. Patients included were of a similar age and had similar vaccination characteristics, but were more frequently men than influenza‐like illness patients reported to the network during the study period. Sixty‐one viruses were detected from 56 of 96 (58%) interpretable samples. The respiratory viruses detected most frequently were metapneumovirus and respiratory syncytial virus. This study showed that virological diagnosis of 18 respiratory viruses can be combined with surveillance of clinical influenza‐like illness in general practice. Although feasibility has not been demonstrated yet, it will be evaluated over the winter of 2010–2011. J. Med. Virol. 83:1451–1457, 2011. © 2011 Wiley‐Liss, Inc.


title: Pregnant Women in Trials of COVID‐19: A Critical Time to Consider Ethical Frameworks of Inclusion in Clinical Trials

text: Ethical issues abound during this unprecedented international public health crisis of Covid‐19. While the trade‐off between societal and individual interests that occurs at the intersection of public health ethics and clinical ethics affects all populations, this calculus has particular relevance for pregnant women and the question of when they will have access to new Covid‐19 therapies and vaccines. Pregnant women are a “scientifically complex” population whose inclusion in clinical research must be done with consideration of the unique state of pregnancy. Yet research on the impact of Covid‐19 on pregnant women is lagging. In a rush to prevent and treat SARS‐CoV‐2 infection, it is crucial that the interests of pregnant women be prioritized to enable them to make autonomous, informed decisions about participating in clinical trials. The global pandemic calls for a revisiting of frameworks for the inclusion of pregnant women in research, as these women have an important stake in the prevention and treatment of Covid‐19.


Aerosol-containment device for prevention of aerosol dispersion during nebulization in COVID-19 patients.


title: Head and neck virtual medicine in a pandemic era: Lessons from COVID‐19

text: The 2019 novel coronavirus disease (COVID‐19) has presented the world and physicians with a unique public health challenge. In light of its high transmissibility and large burden on the health care system, many hospitals and practices have opted to cancel elective surgeries in order to mobilize resources, ration personal protective equipment and guard patients from the virus. Head and neck cancer physicians are particularly affected by these changes given their scope of practice, complex patient population, and interventional focus. In this viewpoint, we discuss some of the many challenges faced by head and neck surgeons in this climate. Additionally, we outline the utility of telemedicine as a potential strategy for allowing physicians to maintain an effective continuum of care.


title: Emergence of a Novel Coronavirus (COVID-19): Protocol for Extending Surveillance Used by the Royal College of General Practitioners Research and Surveillance Centre and Public Health England

text: BACKGROUND: The Royal College of General Practitioners (RCGP) Research and Surveillance Centre (RSC) and Public Health England (PHE) have successfully worked together on the surveillance of influenza and other infectious diseases for over 50 years, including three previous pandemics. With the emergence of the international outbreak of the coronavirus infection (COVID-19), a UK national approach to containment has been established to test people suspected of exposure to COVID-19. At the same time and separately, the RCGP RSC’s surveillance has been extended to monitor the temporal and geographical distribution of COVID-19 infection in the community as well as assess the effectiveness of the containment strategy. OBJECTIVES: The aims of this study are to surveil COVID-19 in both asymptomatic populations and ambulatory cases with respiratory infections, ascertain both the rate and pattern of COVID-19 spread, and assess the effectiveness of the containment policy. METHODS: The RCGP RSC, a network of over 500 general practices in England, extract pseudonymized data weekly. This extended surveillance comprises of five components: (1) Recording in medical records of anyone suspected to have or who has been exposed to COVID-19. Computerized medical records suppliers have within a week of request created new codes to support this. (2) Extension of current virological surveillance and testing people with influenza-like illness or lower respiratory tract infections (LRTI)—with the caveat that people suspected to have or who have been exposed to COVID-19 should be referred to the national containment pathway and not seen in primary care. (3) Serology sample collection across all age groups. This will be an extra blood sample taken from people who are attending their general practice for a scheduled blood test. The 100 general practices currently undertaking annual influenza virology surveillance will be involved in the extended virological and serological surveillance. (4) Collecting convalescent serum samples. (5) Data curation. We have the opportunity to escalate the data extraction to twice weekly if needed. Swabs and sera will be analyzed in PHE reference laboratories. RESULTS: General practice clinical system providers have introduced an emergency new set of clinical codes to support COVID-19 surveillance. Additionally, practices participating in current virology surveillance are now taking samples for COVID-19 surveillance from low-risk patients presenting with LRTIs. Within the first 2 weeks of setup of this surveillance, we have identified 3 cases: 1 through the new coding system, the other 2 through the extended virology sampling. CONCLUSIONS: We have rapidly converted the established national RCGP RSC influenza surveillance system into one that can test the effectiveness of the COVID-19 containment policy. The extended surveillance has already seen the use of new codes with 3 cases reported. Rapid sharing of this protocol should enable scientific critique and shared learning. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/18606


title: Validation of the Fear of COVID-19 Scale in a US College Sample

text: The COVID-19 pandemic has impacted college students’ coursework, stress levels, and perceived health. Various estimates indicate that high proportions of college students have experienced increased amounts of stress (Dziech, Inside Higher Education, 2020; Hartocollis, New York Times, 2020), yet other sources indicate that many college students respond to these changes with resilience (Kelley, Cornell Chronicle, 2020). A method for assessing student anxiety regarding the pandemic is thus needed. The Fear of COVID-19 Scale (FCV-19S) (Ahorsu et al., International Journal of Mental Health and Addiction, 2020) is a seven-item scale which has been validated and shown to possess good psychometric qualities in studies of participants from multiple countries. The current study used a cross-sectional convenience sample of US college student participants (n = 237) and found that the FCV-19S has high reliability and validity as demonstrated by its internal consistency and strong one-factor solution. Scores on the FCV-19S were positively correlated with anxiety for students who were married or of Asian descent. Additionally, the FCV-19S was only moderately correlated with the Generalized Anxiety Disorder-7 Scale (GAD-7), suggesting that the FCV-19S may bring added utility to research and clinical practice with populations impacted by the COVID-19 pandemic.


title: Modulation of cell surface transport and lipid raft localization by the cytoplasmic tail of the influenza virus hemagglutinin

text: Viral glycoproteins are highly variable in their primary structure, but on the other hand feature a high functional conservation to fulfil their versatile tasks during the pathogenic life cycle. Typically, all protein domains are optimized in that indispensable functions can be assigned to small conserved motifs or even individual amino acids. The cytoplasmic tail of many viral spike proteins, although of particular relevance for the virus biology, is often only insufficiently characterized. Hemagglutinin (HA), the receptor‐binding protein of the influenza virus comprises a short cytoplasmic tail of 13 amino acids that exhibits three highly conserved palmitoylation sites. However, the particular importance of these modifications and the tail in general for intracellular trafficking and lateral membrane organization remains elusive. In this study, we generated HA core proteins consisting of transmembrane domain, cytoplasmic tail and a minor part of the ectodomain, tagged with a yellow fluorescent protein. Different mutation and truncation variants of these chimeric proteins were investigated using confocal microscopy, to characterize the role of cytoplasmic tail and palmitoylation for the intracellular trafficking to plasma membrane and Golgi apparatus. In addition, we assessed raft partitioning of the variants by Foerster resonance energy transfer with an established raft marker. We revealed a substantial influence of the cytoplasmic tail length on the intracellular distribution and surface exposure of the proteins. A complete removal of the tail hampers a physiological trafficking of the protein, whereas a partial truncation can be compensated by cytoplasmic palmitoylations. Plasma membrane raft partitioning on the other hand was found to imperatively require palmitoylations, and the cysteine at position 551 turned out to be of most relevance. Our data shed further light on the tight interconnection between cytoplasmic elements and intracellular trafficking and suggest a function of HA palmitoylations in both lateral sorting and anterograde trafficking of the glycoprotein.


title: Genomic and serological detection of bat coronavirus from bats in the Philippines

text: Bat coronavirus (BtCoV) is assumed to be a progenitor of severe acute respiratory syndrome (SARS)-related coronaviruses. To explore the distribution of BtCoVs in the Philippines, we collected 179 bats and detected viral RNA from intestinal or fecal samples by RT-PCR. The overall prevalence of BtCoVs among bats was 29.6 %. Phylogenetic analysis of the partial RNA-dependent RNA polymerase gene suggested that one of the detected BtCoVs was a novel alphacoronavirus, while the others belonged to the genus Betacoronavirus. Western blotting revealed that 66.5 % of bat sera had antibodies to BtCoV. These surveys suggested the endemic presence of BtCoVs in the Philippines.


title: Antigenic and cellular localisation analysis of the severe acute respiratory syndrome coronavirus nucleocapsid protein using monoclonal antibodies

text: Abstract A member of the family of coronaviruses has previously been identified as the cause of the severe acute respiratory syndrome (SARS). In this study, several monoclonal antibodies against the nucleocapsid protein have been generated to examine distribution of the nucleocapsid in virus-infected cells and to study antigenic regions of the protein. Confocal microscopic analysis identified nucleocapsids packaged in vesicles in the perinuclear area indicating viral synthesis at the endoplasmic reticulum and Golgi apparatus. The monoclonal antibodies bound to the central and carboxyterminal half of the nucleocapsid protein indicating prominent exposure and immunogenicity of this part of the protein. Antibodies recognised both linear and conformational epitopes. Predictions of antigenicity using mathematical modelling based on hydrophobicity analysis of SARS nucleoprotein could not be confirmed fully. Antibody binding to discontinuous peptides provides evidence that amino acids 274–283 and 373–382 assemble to a structural unit particularly rich in basic amino acids. In addition, amino acids 286–295, 316–325 and 361–367 that represent the epitope recognised by monoclonal antibody 6D11C1 converge indicating a well-structured C-terminal region of the SARS virus nucleocapsid protein and functional relationship of the peptide regions involved. Alternatively, dimerisation of the nucleocapsid protein may result in juxtaposition of the amino acid sequences 316–325 and 361–367 on one nucleoprotein molecule to amino acid 286–295 on the second peptide. The monoclonal antibodies will be available to assess antigenicity and immunological variabilities between different SARS CoV strains.


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title: Methodology in diagnostic virology

text: Selection of proper assays and appropriate interpretation of results can be a challenge to the veterinary clinician. Assays vary in methodology, sensitivity, and specificity. These assays can be invaluable in attaining the correct diagnosis, but a clear understanding of the assay and the results is essential. To this end, communication with the laboratory personnel is crucial. Optimal sample selection, shipping recommendations, assay selection, and interpretation should be discussed with the laboratory staff.


title: A Review of Quantitative Tools Used to Assess the Epidemiology of Porcine Reproductive and Respiratory Syndrome in U.S. Swine Farms Using Dr. Morrison’s Swine Health Monitoring Program Data

text: Porcine reproductive and respiratory syndrome (PRRS) causes far-reaching financial losses to infected countries and regions, including the U.S. The Dr. Morrison’s Swine Health Monitoring Program (MSHMP) is a voluntary initiative in which producers and veterinarians share sow farm PRRS status weekly to contribute to the understanding, in quantitative terms, of PRRS epidemiological dynamics and, ultimately, to support its control in the U.S. Here, we offer a review of a variety of analytic tools that were applied to MSHMP data to assess disease dynamics in quantitative terms to support the decision-making process for veterinarians and producers. Use of those methods has helped the U.S. swine industry to quantify the cyclical patterns of PRRS, to describe the impact that emerging pathogens has had on that pattern, to identify the nature and extent at which environmental factors (e.g., precipitation or land cover) influence PRRS risk, to identify PRRS virus emerging strains, and to assess the influence that voluntary reporting has on disease control. Results from the numerous studies reviewed here provide important insights into PRRS epidemiology that help to create the foundations for a near real-time prediction of disease risk, and, ultimately, will contribute to support the prevention and control of, arguably, one of the most devastating diseases affecting the North American swine industry. The review also demonstrates how different approaches to analyze and visualize the data may help to add value to the routine collection of surveillance data and support infectious animal disease control.


title: Chapter 17 Antimicrobial lead compounds from marine plants

text: Abstract Marine environment is a home to a very wide diversity of flora and fauna, which includes an array of genetically diverse coastline and under seawater plant species, animal species, microbial species, their habitats, ecosystems, and supporting ecological processes. The Earth is home to an estimated 10million species, of which a large chunk belongs to marine environment. Marine plants are a store house of a variety of antimicrobial compounds like classes of marine flavonoids—flavones and flavonols, terpenoids, alkaloids, peptides, carbohydrates, fatty acids, polyketides, polysaccharides, phenolic compounds, and steroids. Lot of research today is directed toward marine species, which have proved to be a potent source of structurally widely diverse and yet highly bioactive secondary metabolites. Varied species of phylum Porifera, algae including diatoms, Chlorophyta, Euglenophyta, Dinoflagellata, Chrysophyta, cyanobacteria, Rhodophyta, and Phaeophyta, bacteria, fungi, and weeds have been exploited by mankind for their inherent indigenous biological antimicrobial compounds, produced under the extreme stressful underwater conditions of temperature, atmospheric pressure, light, and nutrition. The present study aims at presenting a brief review of bioactive marine compounds possessing antimicrobial potency.


title: ‘Self-collected upper respiratory tract swabs for COVID-19 test’: A feasible way to increase overall testing rate and conserve resources in South Africa

text: Disparity in the testing rate of SARS-CoV-2 amongst different countries and regions is a very big challenge in understanding the COVID-19 pandemic. Although some developed countries have a very high testing rate and subsequently a high number of confirmed cases, less developed countries have a low testing rate and an illusive positivity rate. Collection of the upper respiratory specimen is not often comfortable. The discomfort could be accompanied with epistaxis and headache in some patients. The trained personnel taking the swab is forced to protect self with personal protective equipment (PPE) to avoid infections that may result from the patient due to provoked cough, sneezing and spitting. This study looks into an efficient means of increasing the testing rate for COVID 19 without compromising the quality. A literature review was conducted on the different modalities of collecting upper respiratory specimens and assessing the efficacy of samples collected using different methods in terms of the laboratory yield of different pathogens. Self-collection of upper respiratory tract specimen for diagnostic purposes is not new. Studies have demonstrated that trained staff-collected nasal swabs are not in any way superior to self-collected or parent-assisted swabs. The laboratory yield of different specimens is not determined by who took the sample but by the anatomical site from where the specimen was collected. Self collection of the upper respiratory swabs will not only increase the testing rate but also preserve the scarce PPE and reduces health care worker’s COVID 19 infection rate in South Africa.


Surveys, Serologies, and Sequences Reveal History of Iatrogenic Transmission of HIV-1


title: Mapping of Minimal Motifs of B-Cell Epitopes on Human Zona Pellucida Glycoprotein-3

text: The human zona pellucida glycoprotein-3 (hZP3) by virtue of its critical role during fertilization has been proposed as a promising candidate antigen to develop a contraceptive vaccine. In this direction, it is imperative to map minimal motifs of the B cell epitopes (BCEs) so as to avoid ZP-specific oophoritogenic T cell epitopes (TCEs) in the ZP3-based immunogens. In this study, based on known results of mapping marmoset and bonnet monkey ZP3 (mstZP3 and bmZP3), two predictable epitopes(23–30 and 301–320) on hZP3 were first confirmed and five minimal motifs within four epitopes on hZP3 were defined using serum to recombinant hZP3a(22–176) or hZP3b(177–348) as well as a biosynthetic peptide strategy. These defined minimal motifs were QPLWLL(23–28) for hZP3(23–30), MQVTDD(103–108) for hZP3(93–110), EENW(178–181) for hZP3(172–190), as well as SNSWF(306–310) and EGP(313–315) for hZP3(301–320), respectively. Furthermore, the antigenicity of two peptides for hZP3(172–187) and hZP3(301–315) and specificity of the antibody response to these peptides were also evaluated, which produced high-titer antibodies in immunized animals that were capable of reacting to ZP on human oocytes, r-hZP3b(177–348) protein, as well as r-hZP3(172–190), r-hZP3(303–310), and r-hZP3(313–320) epitope peptides fused with truncated GST188 protein.


title: Dentists thoughts about COVID-19

text: Data sources Self-developed survey. Data extraction and synthesis This was a cross-sectional study using a self-administered online survey. Results Out of 700 randomly selected dentists in Jordan, 368 completed an online survey. Of those, 36% reported that the incubation period for COVID-19 is 1-14 days. The majority were aware of the symptoms of the disease and can identify affected patients. Nonetheless, 36.7% of dentists did not believe it to be a serious public health issue. Conclusions This group of Jordanian dentists was generally aware of COVID-19 symptoms and mode of transmission. Clinical practice guidelines should be made available to dentists by their associations during such crises.


title: A case of SARS‐CoV‐2 infection in an untreated HIV patient in Tokyo, Japan

text: We report a 28‐year‐old male co‐infected with SARS‐CoV‐2 (severe acute respiratory syndrome coronavirus 2; COVID‐19) and human immunodeficiency virus (HIV). His clinical course of the COVID‐19 pneumonia improved smoothly in 9 days; however, his immune status from HIV infection was not well‐controlled due to a lack of antiretroviral therapy (ART). The HIV‐1 viral load decreased during the acute phase and subsequently increased in the recovery phase. Antiretroviral therapy was given a month after the twice non‐detections of SARS‐CoV‐2 to avoid immune reconstitution syndrome (IRIS). Previous reports suggest COVID‐19 do not become more severe in untreated HIV patients than non‐HIV patients. However, untreated HIV patients with COVID‐19 should be followed up carefully for IRIS with ART. This article is protected by copyright. All rights reserved.


title: Computational Determination of Potential Inhibitors of SARS-CoV-2 Main Protease

text: [Image: see text] The novel coronavirus (SARS-CoV-2) has infected several million people and caused thousands of deaths worldwide since December 2019. As the disease is spreading rapidly all over the world, it is urgent to find effective drugs to treat the virus. The main protease (Mpro) of SARS-CoV-2 is one of the potential drug targets. Therefore, in this context, we used rigorous computational methods, including molecular docking, fast pulling of ligand (FPL), and free energy perturbation (FEP), to investigate potential inhibitors of SARS-CoV-2 Mpro. We first tested our approach with three reported inhibitors of SARS-CoV-2 Mpro, and our computational results are in good agreement with the respective experimental data. Subsequently, we applied our approach on a database of ∼4600 natural compounds, as well as 8 available HIV-1 protease (PR) inhibitors and an aza-peptide epoxide. Molecular docking resulted in a short list of 35 natural compounds, which was subsequently refined using the FPL scheme. FPL simulations resulted in five potential inhibitors, including three natural compounds and two available HIV-1 PR inhibitors. Finally, FEP, the most accurate and precise method, was used to determine the absolute binding free energy of these five compounds. FEP results indicate that two natural compounds, cannabisin A and isoacteoside, and an HIV-1 PR inhibitor, darunavir, exhibit a large binding free energy to SARS-CoV-2 Mpro, which is larger than that of 13b, the most reliable SARS-CoV-2 Mpro inhibitor recently reported. The binding free energy largely arises from van der Waals interaction. We also found that Glu166 forms H-bonds to all of the inhibitors. Replacing Glu166 by an alanine residue leads to ∼2.0 kcal/mol decreases in the affinity of darunavir to SARS-CoV-2 Mpro. Our results could contribute to the development of potential drugs inhibiting SARS-CoV-2.


title: Wildlife as Source of Zoonotic Infections

text: Zoonoses with a wildlife reservoir represent a major public health problem, affecting all continents. Hundreds of pathogens and many different transmission modes are involved, and many factors influence the epidemiology of the various zoonoses. The importance and recognition of wildlife as a reservoir of zoonoses are increasing. Cost-effective prevention and control of these zoonoses necessitate an interdisciplinary and holistic approach and international cooperation. Surveillance, laboratory capability, research, training and education, and communication are key elements.


title: The seafood Musculus senhousei shows anti-influenza A virus activity by targeting virion envelope lipids

text: Abstract Marine environments are known to be a new source of structurally diverse bioactive molecules. In this paper, we identified a porphyrin derivative of Pyropheophorbide a (PPa) from the mussel Musculus senhousei (M. senhousei) that showed broad anti-influenza A virus activity in vitro against a panel of influenza A viral strains. The analysis of the mechanism of action indicated that PPa functions in the early stage of virus infection by interacting with the lipid bilayer of the virion, resulting in an alteration of membrane-associated functions, thereby blocking the entry of enveloped viruses into host cells. In addition, the anti-influenza A virus activity of PPa was further assessed in mice infected with the influenza A virus. The survival rate and mean survival time of mice were apparently prolonged compared with the control group which was not treated with the drug. Therefore, PPa and its derivatives may represent lead compounds for controlling influenza A virus infection.


title: NMR assignment of the nonstructural protein nsp3(1066–1181) from SARS-CoV

text: Sequence-specific NMR assignments of the globular core comprising the residues 1066–1181 within the non-structural protein nsp3e from the SARS coronavirus have been obtained using triple-resonance NMR experiments with the uniformly [(13)C,(15)N]-labeled protein. The backbone and side chain assignments are nearly complete, providing the basis for the ongoing NMR structure determination. A preliminary identification of regular secondary structures has been derived from the (13)C chemical shifts.


title: Circulation and genetic diversity of Feline coronavirus type I and II from clinically healthy and FIP‐suspected cats in China

text: Feline infectious peritonitis (FIP) is a fatal infectious disease of wild and domestic cats, and the occurrence of FIP is frequently reported in China. To trace the evolution of type I and II feline coronavirus in China, 115 samples of ascetic fluid from FIP‐suspected cats and 54 fecal samples from clinically healthy cats were collected from veterinary hospitals in China. The presence of FCoV in the samples was detected by RT‐PCR targeting the 6b gene. The results revealed that a total of 126 (74.6%, 126/169) samples were positive for FCoV: 75.7% (87/115) of the FIP‐suspected samples were positive for FCoV, and 72.2% (39/54) of the clinically healthy samples were positive for FCoV. Of the 126 FCoV‐positive samples, 95 partial S genes were successfully sequenced. The partial S gene‐based genotyping indicated that type I FCoV and type II FCoV accounted for 95.8% (91/95) and 4.2% (4/95), respectively. The partial S gene‐based phylogenetic analyses showed that the 91 type I FCoV strains exhibited genetic diversity; the four type II FCoV strains exhibited a close relationship with type II FCoV strains from Taiwan. Three type I FCoV strains, HLJ/HRB/2016/10, HLJ/HRB/2016/11 and HLJ/HRB/2016/13, formed one potential new clade in the nearly complete genome‐based phylogenetic trees. Further analysis revealed that FCoV infection appeared to be significantly correlated with a multi‐cat environment (p < 0.01) and with age (p < 0.01). The S gene of the three type I FCoV strains identified in China, BJ/2017/27, BJ/2018/22 and XM/2018/04, exhibited a six nucleotide deletion (C(4035) AGCTC (4040)). Our data provide evidence that type I and type II FCoV strains co‐circulate in the FIP‐affected cats in China. Type I FCoV strains exhibited high prevalence and genetic diversity in both FIP‐affected cats and clinically healthy cats, and a multi‐cat environment and age (<6 months) were significantly associated with FCoV infection.


title: Reporting of all cardiac medications and their outcome in COVID ‐ 19

text: We read with much interest the article “Organ‐ protective effect of Angiotensin Converting Enzyme 2 and its Effect on the prognosis of COVID ‐19” by Cheng et al in your esteemed Journal. Authors have discussed the theoretical basis of Angiotensin Converting Enzyme 2 in improving outcome of COVID – 19 patients(1). This article is protected by copyright. All rights reserved.


docid: gmzbinpi

title: Molecular Virology of Hepatitis E Virus

text: This review details the molecular virology of the hepatitis E virus (HEV). While replicons and in vitro infection systems have recently become available, a lot of information on HEV has been generated through comparisons with better-studied positive-strand RNA viruses and through subgenomic expression of viral open reading frames. These models are now being verified with replicon and infection systems. We provide here the current knowledge on the HEV genome and its constituent proteins - ORF1, ORF2 and ORF3. Based on the available information, we also modify the existing model of the HEV life cycle.


Within a large healthcare system, the incidence of positive COVID-19 results and mortality are lower in patients on chronic hydroxychloroquine therapy


title: Cutting Cubes Out of Fog: The Whole View of Design

text: Abstract In this article we argue that the field of design evolved to include helping organizations address complex problems in the face of uncertainty, but it has yet to build knowledge in a way that formalizes the various activities embraced by the field. We use our relationship with the school of public health at Harvard to illustrate the advantages of formalizing design knowledge more explicitly. We introduce the Whole View model to provide the field of design with a tentative structure for its knowledge that could make it easier for design and other fields to work together. We situate the model in the context of different types of design education, and show examples of how the Whole View provides a structure for teaching, research, and practice. We conclude by introducing a new agenda for design centered on the well-being of people, organizations, and the natural environment, with health, happiness, and prosperity as the levers driving change.


title: Detecting the emergence of novel, zoonotic viruses pathogenic to humans

text: RNA viruses, with their high potential for mutation and epidemic spread, are the most common class of pathogens found as new causes of human illness. Despite great advances made in diagnostic technology since the 1950s, the annual rate at which novel virulent viruses have been found has remained at 2–3. Most emerging viruses are zoonoses; they have jumped from mammal or bird hosts to humans. An analysis of virus discovery indicates that the small number of novel viruses discovered annually is an artifact of inadequate surveillance in tropical and subtropical countries, where even established endemic pathogens are often misdiagnosed. Many of the emerging viruses of the future are already infecting humans but remain to be uncovered by a strategy of disease surveillance in selected populations.


title: Association of sputum microbiota profiles with severity of community-acquired pneumonia in children

text: BACKGROUND: Competitive interactions among bacteria in the respiratory tract microbiota influence which species can colonize and potentially contribute to pathogenesis of community-acquired pneumonia (CAP). However, understanding of the role of respiratory tract microbiota in the clinical course of pediatric CAP is limited. METHODS: We sought to compare microbiota profiles in induced sputum and nasopharyngeal/oropharyngeal (NP/OP) samples from children and to identify microbiota profiles associated with CAP severity. We used 16S ribosomal RNA sequencing and several measures of microbiota profiles, including principal component analysis (PCA), to describe the respiratory microbiota in 383 children, 6 months to <18 years, hospitalized with CAP. We examined associations between induced sputum and NP/OP microbiota profiles and CAP severity (hospital length of stay and intensive care unit admission) using logistic regression. RESULTS: Relative abundance of bacterial taxa differed in induced sputum and NP/OP samples. In children 6 months to < 5 years, the sputum PCA factor with high relative abundance of Actinomyces, Veillonella, Rothia, and Lactobacillales was associated with decreased odds of length of stay ≥ 4 days [adjusted odds ratio (aOR) 0.69; 95 % confidence interval (CI) 0.48–0.99]. The sputum factor with high relative abundance of Haemophilus and Pasteurellaceae was associated with increased odds of intensive care unit admission [aOR 1.52; 95 % CI 1.02–2.26]. In children 5 to < 18 years, the sputum factor with high relative abundance of Porphyromonadaceae, Bacteriodales, Lactobacillales, and Prevotella was associated with increased odds of length of stay ≥ 4 days [aOR 1.52; 95 % CI 1.02–2.26]. Taxa in NP/OP samples were not associated with CAP severity. CONCLUSION: Certain taxa in the respiratory microbiota, which were detected in induced sputum samples, are associated with the clinical course of CAP. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1670-4) contains supplementary material, which is available to authorized users.


title: Pseudorabies virus infection (Aujeszky’s disease) in an Iberian lynx (Lynx pardinus) in Spain: a case report

text: BACKGROUND: The only natural hosts of Pseudorabies virus (PRV) are members of the family Suidae (Sus scrofa scrofa). In species other than suids infection is normally fatal. In these mammals, including carnivores, PRV typically causes serious neurologic disease. The endangered Iberian lynx (Lynx pardinus) is a wild feline endemic to south-western Europe (Iberian Peninsula). The Iberian lynx was found to be the world’s most endangered felid species in 2002. In wild felines, PRV infection has only been previously reported once in a Florida panther in 1994. No seropositive lynxes have ever been found, nor has PRV been detected in dead Iberian lynxes to date. CASE PRESENTATION: We describe the first reported case of pseudorabies in an Iberian lynx (Lynx pardinus). Pseudorabies was diagnosed in a young wild Iberian lynx from Extremadura (SW Spain) by histopathological examination, immunohistochemistry, polymerase chain reaction (PCR) and sequence analysis. Gross lesions included alopecia of the ventral neck, bloody gastro-intestinal contents and congestion of the brain. Histopathological analysis showed a moderate nonsuppurative meningoencephalitis with diffuse areas of demyelination, necrotizing gastritis and enteritis of the small intestine. Pseudorabies virus (PRV) antigen was found in neuronal and non-neuronal cells of the brain, tonsils, and gastric glandular epithelial cells by immunohistochemical analysis. The presence of the virus in the brain was confirmed by nested PCR. The sequence analysis of the 146 bp fragment (from the viral glycoprotein B gene) showed that the amplified sequence matched (with 100% identity) the PRV genome. Furthermore, specific DNA from glycoprotein D and E encoding-genes was detected by conventional and real-time PCR, respectively, confirming the latter that this infection was produced by a wild-type PRV strain. CONCLUSIONS: This study supports the suspicion that PRV could infect the Iberian lynx. The detection of PRV in a dead Iberian lynx suggests that the virus may have a negative impact on the survival of endangered lynxes in the wild. However, because this is the first verified instance of lynx mortality resulting from pseudorabies, its true impact on the population is unknown. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-016-0938-7) contains supplementary material, which is available to authorized users.


docid: aaewccao

title: The respiratory health of urban indigenous children aged less than 5 years: study protocol for a prospective cohort study

text: BACKGROUND: Despite the burden of acute respiratory illnesses (ARI) among Aboriginal and Torres Strait Islander children being a substantial cause of childhood morbidity and associated costs to families, communities and the health system, data on disease burden in urban children are lacking. Consequently evidence-based decision-making, data management guidelines, health resourcing for primary health care services and prevention strategies are lacking. This study aims to comprehensively describe the epidemiology, impact and outcomes of ARI in urban Aboriginal and Torres Strait Islander children (hereafter referred to as Indigenous) in the greater Brisbane area. METHODS/DESIGN: An ongoing prospective cohort study of Indigenous children aged less than five years registered with a primary health care service in Northern Brisbane, Queensland, Australia. Children are recruited at time of presentation to the service for any reason. Demographic, epidemiological, risk factor, microbiological, economic and clinical data are collected at enrolment. Enrolled children are followed for 12 months during which time ARI events, changes in child characteristics over time and monthly nasal swabs are collected. Children who develop an ARI with cough as a symptom during the study period are more intensely followed-up for 28 (±3) days including weekly nasal swabs and parent completed cough diary cards. Children with persistent cough at day 28 post-ARI are reviewed by a paediatrician. DISCUSSION: Our study will be one of the first to comprehensively evaluate the natural history, epidemiology, aetiology, economic impact and outcomes of ARIs in this population. The results will inform studies for the development of evidence-based guidelines to improve the early detection, prevention and management of chronic cough and setting of priorities in children during and after ARI. TRIAL REGISTRATION: Australia New Zealand Clinical Trial Registry Registration Number: 12614001214628. Registered 18 November 2014


title: Transforming a research institute into a COVID diagnostic centre – scientists step forward to protect public health during the coronavirus pandemic

text: Let's face it: COVID‐19 caught us unprepared. Virologists had abundantly described the mutagenic nature of viruses and their potential infectivity. And yet, mankind was surprised by the sudden appearance of SARS‐CoV‐2. We saw what happened in China and watched the coronavirus spread through countries, but we were slow in reacting.


docid: onzzpkye

title: Critically ill COVID-19 patient

text: Coronavirus disease 2019 (COVID-19) stands out as the major pandemic that we have experienced in the last century. As it affects every social structure, it brought the importance of intensive care support once again to the agenda of healthcare system after causing severe acute respiratory syndrome. The precautions to be taken against this virus, where our knowledge is extremely small, intensive care units take an indispensable place in pandemic planning. In this review, we aimed to emphasize the crucial points regarding intensive care management of COVID-19 patients, which we have written not only for intensivists but also for all healthcare professionals.


L’après Covid-19: vers une dermatologie nouvelle?


AEP: Engine of the specialist's recognition and guarantor of the paediatric model. Generating health also in pandemic situations()


title: Effects of 1,2,4,6-tetra-O-galloyl-β-D-glucose from P. emblica on HBsAg and HBeAg secretion in HepG2.2.15 cell culture

text: A polyphenolic compound, 1,2,4,6-tetra-O-galloyl-β-D-glucose (1246TGG), was isolated from the traditional Chinese medicine Phyllanthus emblica L. (Euphorbiaceae) and assayed for its potential as an anti-hepatitis B virus (HBV) agent. The cytotoxicity of 1246TGG on HepG2.2.15 as well as HepG2 cells was determined by observing cytopathic effects, and the effects of 1246TGG on secretion of HBsAg and HBeAg in HepG2.2.15 cells were assayed by enzyme immunoassay. Results indicates that treatment with 1246TGG (6.25 μg/mL, 3.13 μg/mL), reduced both HBsAg and HBeAg levels in culture supernatant, yet the inhibitory effects tend to decline with the assay time. This study provides a basis for further investigation of the anti-HBV activity and possible mechanism of action of 1246TGG.


title: Structural analysis of the putative SARS-CoV-2 primase complex

text: We report the crystal structure of the SARS-CoV-2 putative primase composed of the nsp7 and nsp8 proteins. We observed a dimer of dimers (2:2 nsp7-nsp8) in the crystallographic asymmetric unit. The structure revealed a fold with a helical core of the heterotetramer formed by both nsp7 and nsp8 that is flanked with two symmetry-related nsp8 β-sheet subdomains. It was also revealed that two hydrophobic interfaces one of approx. 1340 Å(2) connects the nsp7 to nsp8 and a second one of approx. 950 Å(2) connects the dimers and form the observed heterotetramer. Interestingly, analysis of the surface electrostatic potential revealed a putative RNA binding site that is formed only within the heterotetramer.


title: Impact of COVID-19 outbreak measures of lockdown on the Italian Carbon Footprint

text: Abstract Stringent lockdown measures implemented in Italy to mitigate the spread of COVID-19 are generating unprecedented economic impacts. However, the environmental consequences associated with the temporary shutdown and recovery of industrial and commercial activities are still not fully understood. Using the well-known carbon footprint (CF) indicator, this paper provides a comprehensive estimation of environmental effects due to the COVID-19 outbreak lockdown measures in Italy. Our aim was to quantify the CF associated with the consumption of energy by any economic activity and region in Italy during the lockdown, and then compare these environmental burdens with the CF calculated for analogous periods from 2015 to 2019 (~March and April). Complementarily, we also conducted a scenario analysis to estimate the post-lockdown CF impact in Italy. A consumption-based approach was applied according to the principles of the established Life Cycle Assessment method. The CF was therefore quantified as a sum of direct and indirect greenhouse gases (GHGs) released from domestically produced and imported energy metabolism flows, excluding the exports. Our findings indicate that the CF in the lockdown period is ~−20% lower than the mean CF calculated for the past. This means avoided GHGs in between ~5.6 and ~10.6 Mt CO2e. Results further suggest that a tendency occurs towards higher impact savings in the Northern regions, on average ~230 kt CO2e of GHGs avoided by province (against ~110–130 kt CO2e in central and Southern provinces). Not surprisingly, these are the utmost industrialized areas of Italy and have been the ones mostly affected by the outbreak. Despite our CF estimates are not free of uncertainties, our research offers quantitative insights to start understanding the magnitude generated by such an exceptional lockdown event in Italy on climate change, and to complement current scientific efforts investigating the relationships between air pollution and the spread of COVID-19.


title: DNA Vaccine Delivered by a Needle-Free Injection Device Improves Potency of Priming for Antibody and CD8+ T-Cell Responses after rAd5 Boost in a Randomized Clinical Trial

text: BACKGROUND: DNA vaccine immunogenicity has been limited by inefficient delivery. Needle-free delivery of DNA using a CO(2)-powered Biojector® device was compared to delivery by needle and syringe and evaluated for safety and immunogenicity. METHODS: Forty adults, 18–50 years, were randomly assigned to intramuscular (IM) vaccinations with DNA vaccine, VRC-HIVDNA016-00-VP, (weeks 0, 4, 8) by Biojector® 2000™ or needle and syringe (N/S) and boosted IM at week 24 with VRC-HIVADV014-00-VP (rAd5) with N/S at 10(10) or 10(11) particle units (PU). Equal numbers per assigned schedule had low (≤500) or high (>500) reciprocal titers of preexisting Ad5 neutralizing antibody. RESULTS: 120 DNA and 39 rAd5 injections were given; 36 subjects completed follow-up research sample collections. IFN-γ ELISpot response rates were 17/19 (89%) for Biojector® and 13/17 (76%) for N/S delivery at Week 28 (4 weeks post rAd5 boost). The magnitude of ELISpot response was about 3-fold higher in Biojector® compared to N/S groups. Similar effects on response rates and magnitude were observed for CD8+, but not CD4+ T-cell responses by ICS. Env-specific antibody responses were about 10-fold higher in Biojector-primed subjects. CONCLUSIONS: DNA vaccination by Biojector® was well-tolerated and compared to needle injection, primed for greater IFN-γ ELISpot, CD8+ T-cell, and antibody responses after rAd5 boosting. TRIAL REGISTRATION: ClinicalTrials.gov NCT00109629


Editorial: COVID-19 and what it means for end-of-life care in ICU: Balancing the priorities


title: A molecular docking model of SARS-CoV S1 protein in complex with its receptor, human ACE2

text: Abstract The exact residues within severe acute respiratory syndrome coronavirus (SARS-CoV) S1 protein and its receptor, human ACE2, involved in their interaction still remain largely undetermined. Identification of exact amino acid residues that are crucial for the interaction of S1 with ACE2 could provide working hypotheses for experimental studies and might be helpful for the development of antiviral inhibitor. In this paper, a molecular docking model of SARS-CoV S1 protein in complex with human ACE2 was constructed. The interacting residue pairs within this complex model and their contact types were also identified. Our model, supported by significant biochemical evidence, suggested receptor-binding residues were concentrated in two segments of S1 protein. In contrast, the interfacial residues in ACE2, though close to each other in tertiary structure, were found to be widely scattered in the primary sequence. In particular, the S1 residue ARG453 and ACE2 residue LYS341 might be the key residues in the complex formation.


title: Respiratory viral coinfection among hospitalized patients with H1N1 2009 during the first pandemic wave in Brazil

text: Abstract Influenza A coinfections with other respiratory viruses were investigated in 25.8% (41/159) of the samples from patients hospitalized in 2009 at our University Hospital. Out of the 41 influenza A cases, nine cases (21.9%) were coinfected with other viruses, with a similar frequency among children and adults (p=0.47), and seasonal influenza cases were more prevalent than H1N1 2009 influenza virus. Adenovirus was the most frequently detected (4/9) among coinfected cases. Coinfection was not associated with higher morbidity or mortality (p=0.75).


title: Biocatalytic properties of a novel crude glycyrrhizin hydrolase from the liver of the domestic duck

text: A novel crude glycyrrhizin (GL) hydrolase preparation from the liver of domestic duck was used to produce glycyrrhetic acid monoglucuronide. To characterize the biocatalytic profiles of the crude enzyme, some effect factors were investigated. It had an apparent optimal pH of 6.0 and an optimal temperature at 55 °C. Most of the metal ions tested and ethylene diamine tetra acetic acid showed little effect on the crude enzyme activity except Cu(2+). The enzyme was stable only at pH 6. It was more prone to inactivity at high pH conditions than at low pH conditions. It was stable at temperatures below 55 °C and it will lost 90% GL hydrolytic activity exposed at 70 °C. GL hydrolytic activity declined by 30% compared with the control in aqueous solution (buffer pH 6.0) when pre-equilibrated at 55 °C for 5 days. It indicated that the novel crude GL hydrolase preparation had good biocatalytic ability for selective hydrolysis of one glucuronic acid from GL.


title: Explosion in mortality in the Amazonian epicenter of the COVID-19 epidemic 19

text: Manaus, the capital of the Brazilian State of Amazonas, is the current epicenter of the COVID-19 epidemic in Amazonia The sharp increase in deaths is a huge concern for health system administrators and society The study aimed to analyze excess overall mortality according to Epidemiological Week (EW) in order to identify changes potentially associated with the epidemic in Manaus Overall and cause-specific mortality data were obtained from the Central Database of the National Civil Registry and the Mortality Information System for 2018, 2019, and 2020 The study analyzed age bracket, sex, place of death, EW, calendar year, and causes of death Ratios were calculated between deaths in 2019/2018 and 2020/2019 to estimate excess deaths, with 5% confidence intervals No significant excess overall mortality was seen in the ratios for 2019/2018, independently of EW Meanwhile, the ratios for 2020/2019 increased from 1 0 (95%CI: 0 9-1 3) in EW 11 to 4 6 (95%CI: 3 9-5 3) in EW 17 Excess overall mortality was observed with increasing age, especially in individuals 60 years or older, who accounted for 69 1% (95%CI: 66 8-71 4) of the deaths The ratios for 2020/2019 for deaths at home or on public byways were 1 1 (95%CI: 0 7-1 8) in EW 12 and 7 8 (95%CI: 5 4-11 2) in EW 17 The explosion in overall mortality in Manaus and the high proportion of deaths at home or on public byways reveals the epidemic's severity in contexts of heavy social inequality and weak effectiveness of government policies, especially policies meant to deal with social inequalities and strengthen the Unified Health System


title: Establishment of a COVID‐19 Recovery Unit in a Veteran Affairs (VA) Post‐Acute Facility

text: Coronavirus disease 19 (COVID‐19) is now an epidemic of global proportion with major adverse impacts on older adults, persons with chronic diseases, and especially residents of long‐term care facilities. This health catastrophe has challenged healthcare facilities’ capacity to deliver care to not only COVID‐19 patients but all patients who need hospital care. We report on a novel approach of utilizing long‐term care beds at a Department of Veterans Affairs healthcare facility for managing recovering COVID‐19 patients.


title: Concurrent detection of other respiratory viruses in children shedding viable human respiratory syncytial virus

text: Human respiratory syncytial virus (HRSV) is an important cause of respiratory disease. The majority of studies addressing the importance of virus co‐infections to the HRSV‐disease have been based on the detection of HRSV by RT‐PCR, which may not distinguish current replication from prolonged shedding of remnant RNA from previous HRSV infections. To assess whether co‐detections of other common respiratory viruses are associated with increased severity of HRSV illnesses from patients who were shedding viable‐HRSV, nasopharyngeal aspirates from children younger than 5 years who sought medical care for respiratory infections in Ribeirão Preto (Brazil) were tested for HRSV by immunofluorescence, RT‐PCR and virus isolation in cell culture. All samples with viable‐HRSV were tested further by PCR for other respiratory viruses. HRSV‐disease severity was assessed by a clinical score scale. A total of 266 samples from 247 children were collected and 111 (42%) were HRSV‐positive. HRSV was isolated from 70 (63%), and 52 (74%) of them were positive for at least one additional virus. HRSV‐positive diseases were more severe than HRSV‐negative ones, but there was no difference in disease severity between patients with viable‐HRSV and those HRSV‐positives by RT‐PCR. Co‐detection of other viruses did not correlate with increased disease severity. HRSV isolation in cell culture does not seem to be superior to RT‐PCR to distinguish infections associated with HRSV replication in studies of clinical impact of HRSV. A high rate of co‐detection of other respiratory viruses was found in samples with viable‐HRSV, but this was not associated with more severe HRSV infection. J Med. Virol. 85:1852–1859, 2013. © 2013 Wiley Periodicals, Inc.


docid: qnxsvurv

title: SARS‐CoV‐2 coinfections: Could influenza and the common cold be beneficial?

text: The novel coronavirus SARS‐CoV‐2 has rapidly spread around the world, causing serious illness and death and creating a heavy burden on the health care systems of many countries. Since the virus first emerged in late November 2019, its spread has coincided with peak circulation of several seasonal respiratory viruses, yet some studies have noted limited coinfections between SARS‐CoV‐2 and other viruses. We use a mathematical model of viral coinfection to study SARS‐CoV‐2 coinfections, finding that SARS‐CoV‐2 replication is easily suppressed by many common respiratory viruses. According to our model, this suppression is because SARS‐CoV‐2 has a lower growth rate (1.8 /d) than the other viruses examined in this study. The suppression of SARS‐CoV‐2 by other pathogens could have implications for the timing and severity of a second wave. This article is protected by copyright. All rights reserved.


A simple and effective protective shield for the ophthalmoscope to prevent COVID‐19


docid: rkrkofri

title: Bat and virus

text: Bat, the only flying mammal and count more than 20% of the extant mammals on earth, were recently identified as a natural reservoir of emerging and reemerging infectious pathogens. Astonishing amount (more than 70) and genetic diversity of viruses isolated from the bat have been identified in different populations throughout the world. Many studies focus on bat viruses that caused severe domestic and human diseases. However, many viruses were found in apparently healthy bats, suggesting that bats may have a specific immune system or antiviral activity against virus infections. Therefore, basic researches for bat immunology and virus-host interactions are important for understanding bat-derived infectious diseases.


title: A facile inhibitor screening of SARS coronavirus N protein using nanoparticle-based RNA oligonucleotide

text: Hundreds of million people worldwide have been infected with severe acute respiratory syndrome (SARS), and the rate of global death from SARS has remarkably increased. Hence, the development of efficient drug treatments for the biological effects of SARS is highly needed. We have previously shown that quantum dots (QDs)-conjugated RNA oligonucleotide is sensitive to the specific recognition of the SARS-associated coronavirus (SARS-CoV) nucleocapsid (N) protein. In this study, we found that a designed biochip could analyze inhibitors of the SARS-CoV N protein using nanoparticle-based RNA oligonucleotide. Among the polyphenolic compounds examined, (−)-catechin gallate and (−)-gallocatechin gallate demonstrated a remarkable inhibition activity on SARS-CoV N protein. (−)-catechin gallate and (−)-gallocatechin gallate attenuated the binding affinity in a concentrated manner as evidenced by QDs-conjugated RNA oligonucleotide on a designed biochip. At a concentration of 0.05 μg mL(−1), (−)-catechin gallate and (−)-gallocatechin gallate showed more than 40% inhibition activity on a nanoparticle-based RNA oligonucleotide biochip system.


title: Glutamyl Endopeptidases: The Puzzle of Substrate Specificity

text: Glutamyl endopeptidases (GEPases) are chymotrypsin-like enzymes that preferentially cleave the peptide bonds of the α-carboxyl groups of glutamic acid. Despite the many years of research, the structural determinants underlying the strong substrate specificity of GEPases still remain unclear. In this review, data concerning the molecular mechanisms that determine the substrate preference of GEPases is generalized. In addition, the biological functions of and modern trends in the research into these enzymes are outlined.


title: Devil and angel in the renin–angiotensin system: ACE–angiotensin II–AT(1) receptor axis vs. ACE2–angiotensin-(1–7)–Mas receptor axis

text: Recent studies have established a new regulatory axis in the renin–angiotensin system (RAS). In this axis, angiotensin (Ang)-(1–7) is finally produced from Ang I or Ang II by the catalytic activity of angiotensin-converting enzyme 2 (ACE2). Ang-(1–7) shows actions different from those of AT(1) receptor stimulation, such as vasodilatation, natriuresis, anti-proliferation and an increase in the bradykinin–NO (nitric oxide) system. As the catalytic efficiency of ACE2 is approximately 400-fold higher with Ang II as a substrate than with Ang I, this axis is possibly acting as a counter-regulatory system against the ACE/Ang II/AT(1) receptor axis. The signaling pathway of the ACE2–Ang-(1–7) axis has not yet been totally and clearly understood. However, a recent report suggests that the Mas oncogene acts as a receptor for Ang-(1–7). Intracellular signaling through Mas is not clear yet. Several factors such as Akt phosphorylation, protein kinase C activation and mitogen-activated protein (MAP) kinase inhibition seem to be involved in this signaling pathway. Further investigations are needed to clarify the regulation and mechanism of action of ACE2 and Ang-(1–7). However, this second axis through ACE2 and Ang-(1–7) in RAS can be an important target for the therapy of cardiovascular and metabolic disorders.


title: Characteristics and Injury Patterns in Electric-Scooter Related Accidents—A Prospective Two-Center Report from Germany

text: Since the introduction of rental E-scooters in Germany in mid-June 2019, the safety of this new means of transport has been the subject of extensive public debate. However, valid data on injuries and usage habits are not yet available. This retrospective two-center study included a total of 76 patients who presented to the emergency department following E-scooter-related accidents. The mean age was 34.3 ± 12.4 years and 69.7% of the patients were male. About half of the patients were admitted by ambulance (42.1%). Fractures were found in 48.6% of patients, and 27.6% required surgical treatment due to a fracture. The upper extremities were the most commonly affected body region, followed by injuries to the lower extremity and to the head and face. Only one patient had worn a helmet. In-hospital treatment was necessary for 26.3% of the cases. Patients presented to the emergency department mainly during the weekend and on-call times. This is the first report on E-scooter-related injuries in Germany. Accidents with E-scooters can cause serious injuries and, therefore, represent a further burden to emergency departments. The use of E-scooters appears to be mostly recreational, and the rate of use of protective gear is low.


title: Surveillance of Infectious Diseases

text: Surveillance is the ongoing, systematic collection, analysis, and interpretation of data essential to public health practice for public health action, closely integrated with the timely dissemination of these data to those who need to know. Modern surveillance systems must meet the challenges of a dynamic emergence of infectious diseases and the factors that influence it, including human demographics and behavior, technology, economic development and land use, international travel and commerce, microbial adaptation and change, breakdown of public health measures, and potential acts of bioterrorism. Globally, the use of public health surveillance systems to track occurrences of infectious diseases and other public health threats has never been more important, and all countries have agreed to commit to conduct surveillance under the World Health Organization International Health Regulations (2005). Modern information technology can facilitate linking diverse sources of data, rapid analysis and synthesis of large amounts of information, and development of the communications and networks needed globally for surveillance and response to infectious diseases. Continued development of diagnostic and molecular laboratory methods, when effectively combined with epidemiologic information, can make prevention, recognition, and response to outbreaks faster and more effective.


title: Selection of reference genes for gene expression analysis by real-time qPCR in avian cells infected with infectious bronchitis virus.

text: Infectious bronchitis virus (IBV) causes infectious bronchitis in poultry, a respiratory disease that is a source of major economic loss to the poultry industry. Detection and the study of the molecular pathogenesis of the virus often involve the use of real-time quantitative PCR assays (qPCR). To account for error within the experiments, the levels of target gene transcription are normalized to that of suitable reference genes. Despite publication of the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines in 2009, single un-tested reference genes are often used for normalization of qPCR assays in avian research studies. Here, we use the geNorm algorithm to identify suitable reference genes in different avian cell types during infection with apathogenic and pathogenic strains of IBV. We discuss the importance of selecting an appropriate experimental sample subset for geNorm analysis, and show the effect that this selection can have on resultant reference gene selection. The effects of inappropriate normalization on the transcription pattern of a cellular signalling gene, AKT1, and the interferon-inducible, MX1, were studied. We identify the possibility of the misinterpretation of qPCR data when an inappropriate normalization strategy is employed. This is most notable when measuring the transcription of AKT1, where changes are minimal during infection.


“Does having diabetes increase chances of contracting COVID-19 infection?”


title: Common Respiratory Tract Infections as Psychological Entities: A Review of the Mood and Performance Effects of Being Ill

text: The clinical manifestations associated with colds and influenza overshadow the equally important mood and performance impairments. While decreased alertness and increased anxiety can be considered side effects of symptomatology, symptoms alone may not be responsible for the psychomotor and attention deficits of colds and influenza, respectively. An alternative hypothesis, as proposed in this review, suggests that the immune response, in the form of a cytokine cascade, may be responsible for both the physical and psychological symptoms. In particular, patterns of cytokine production for each infection will dictate the symptoms and performance deficits both within and between viruses. This hypothesis can be extended to incorporate infectious mononucleosis, as well as colds and influenza. The efficacy of symptom‐based overthe‐counter medications is then called into question.


docid: lavcsqov

title: Human Coronaviruses and Other Respiratory Viruses: Underestimated Opportunistic Pathogens of the Central Nervous System?

text: Respiratory viruses infect the human upper respiratory tract, mostly causing mild diseases. However, in vulnerable populations, such as newborns, infants, the elderly and immune-compromised individuals, these opportunistic pathogens can also affect the lower respiratory tract, causing a more severe disease (e.g., pneumonia). Respiratory viruses can also exacerbate asthma and lead to various types of respiratory distress syndromes. Furthermore, as they can adapt fast and cross the species barrier, some of these pathogens, like influenza A and SARS-CoV, have occasionally caused epidemics or pandemics, and were associated with more serious clinical diseases and even mortality. For a few decades now, data reported in the scientific literature has also demonstrated that several respiratory viruses have neuroinvasive capacities, since they can spread from the respiratory tract to the central nervous system (CNS). Viruses infecting human CNS cells could then cause different types of encephalopathy, including encephalitis, and long-term neurological diseases. Like other well-recognized neuroinvasive human viruses, respiratory viruses may damage the CNS as a result of misdirected host immune responses that could be associated with autoimmunity in susceptible individuals (virus-induced neuro-immunopathology) and/or viral replication, which directly causes damage to CNS cells (virus-induced neuropathology). The etiological agent of several neurological disorders remains unidentified. Opportunistic human respiratory pathogens could be associated with the triggering or the exacerbation of these disorders whose etiology remains poorly understood. Herein, we present a global portrait of some of the most prevalent or emerging human respiratory viruses that have been associated with possible pathogenic processes in CNS infection, with a special emphasis on human coronaviruses.


Covid-19: Major US medical organisations urge people to wear masks.


title: Nanoparticulate vacuolar ATPase blocker exhibits potent host-targeted antiviral activity against feline coronavirus

text: Feline infectious peritonitis (FIP), caused by a mutated feline coronavirus, is one of the most serious and fatal viral diseases in cats. The disease remains incurable, and there is no effective vaccine available. In light of the pathogenic mechanism of feline coronavirus that relies on endosomal acidification for cytoplasmic entry, a novel vacuolar ATPase blocker, diphyllin, and its nanoformulation are herein investigated for their antiviral activity against the type II feline infectious peritonitis virus (FIPV). Experimental results show that diphyllin dose-dependently inhibits endosomal acidification in fcwf-4 cells, alters the cellular susceptibility to FIPV, and inhibits the downstream virus replication. In addition, diphyllin delivered by polymeric nanoparticles consisting of poly(ethylene glycol)-block-poly(lactide-co-glycolide) (PEG-PLGA) further demonstrates an improved safety profile and enhanced inhibitory activity against FIPV. In an in vitro model of antibody-dependent enhancement of FIPV infection, diphyllin nanoparticles showed a prominent antiviral effect against the feline coronavirus. In addition, the diphyllin nanoparticles were well tolerated in mice following high-dose intravenous administration. This study highlights the therapeutic potential of diphyllin and its nanoformulation for the treatment of FIP.


title: Selected TLR7/8 agonist and type I interferon (IFN-α) cooperatively redefine the microglia transcriptome

text: BACKGROUND: Microglia, the primary immune cells of the central nervous system, exerts multiple functions to mediate many neurological diseases. Upon any detection of invading pathogen products (e.g., TLR agonists) or host-released signaling factors (e.g., interferon/IFN), these cells undergo an activation process to release large numbers of inflammatory substances that participate in inflammation and homeostasis. The profound effects of inflammation associated with TLR7/8 agonist Resiquimod (R848) and type 1 interferon (e.g., IFN-α)-induced macrophage and dendritic cell activation on biological outcomes have long been recognized. However, the underlying mechanisms are not well defined in microglial cells. METHODS: The present study investigated the molecular signatures of microglia and identified genes that are uniquely or synergistically expressed in R848-, IFN-α- or R848 with IFN-α-treated primary microglial (PM) cells. We used RNA-sequencing, quantitative real-time PCR, and bioinformatics approaches to derive regulatory networks that control the transcriptional response of PM to R848, IFN-α and R848 with IFN-α. RESULTS: Our approach revealed that the inflammatory response in R848 with IFN-α-treated PM is faster and more intense than that in R848 or IFN-α-treated PM in terms of the number of differentially expressed genes and the magnitude of induction/repression. In particular, our integrative analysis enabled us to suggest the regulatory functions of TFs, which allowed the construction of a network model that explains how TLR7/8 and IFN-α-sensing pathways achieve specificity. CONCLUSION: In conclusion, the systematic approach presented herein could be important to the understanding microglial activation-mediated molecular signatures induced by inflammatory stimuli related to TLR7/8, IFN-α or co-signaling, and associated transcriptional machinery of microglial functions and neuroinflammatory mechanisms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10787-019-00610-8) contains supplementary material, which is available to authorized users.


title: Single particle assay of coronavirus membrane fusion with proteinaceous receptor-embedded supported bilayers

text: Abstract Total internal reflection microscopy combined with microfluidics and supported bilayers is a powerful, single particle tracking (SPT) platform for host-pathogen membrane fusion studies. But one major inadequacy of this platform has been capturing the complexity of the cell membrane, including membrane proteins. Because of this, viruses requiring proteinaceous receptors, or other unknown cellular co-factors, have been precluded from study. Here we describe a general method to integrate proteinaceous receptors and cellular components into supported bilayers for SPT fusion studies. This method is general to any enveloped virus-host cell pair, but demonstrated here for feline coronavirus (FCoV). Supported bilayers are formed from mammalian cell membrane vesicles that express feline aminopeptidase N (the viral receptor) using a cell blebbing technique. SPT is then used to identify fusion intermediates and measure membrane fusion kinetics for FCoV. Overall, the fusion results recapitulate what is observed in vivo, that coronavirus entry requires binding to specific receptors, a low-pH environment, and that membrane fusion is receptor- and protease-dependent. But this method also provides quantitative kinetic rate parameters for intermediate steps in the coronavirus fusion pathway, which to our knowledge have not been obtained before. Moreover, the platform offers versatile, precise control over the sequence of triggers for fusion; these triggers may define the fusion pathway, tissue tropism, and pathogenicity of coronaviruses. Systematically varying these triggers in this platform provides a new route to study how viruses rapidly adapt to other hosts, and to identify factors that led to the emergence of zoonotic viruses, such as human SARS-CoV and the newly emerging human MERS-CoV.


title: Responding to the Pandemic of Falsified Medicines

text: Over the past decade, the number of countries reporting falsified (fake, spurious/falsely labeled/counterfeit) medicines and the types and quantities of fraudulent drugs being distributed have increased greatly. The obstacles in combating falsified pharmaceuticals include 1) lack of consensus on definitions, 2) paucity of reliable and scalable technology to detect fakes before they reach patients, 3) poor global and national leadership and accountability systems for combating this scourge, and 4) deficient manufacturing and regulatory challenges, especially in China and India where fake products often originate. The major needs to improve the quality of the world's medicines fall into three main areas: 1) research to develop and compare accurate and affordable tools to identify high-quality drugs at all levels of distribution; 2) an international convention and national legislation to facilitate production and utilization of high-quality drugs and protect all countries from the criminal and the negligent who make, distribute, and sell life-threatening products; and 3) a highly qualified, well-supported international science and public health organization that will establish standards, drug-quality surveillance, and training programs like the U.S. Food and Drug Administration. Such leadership would give authoritative guidance for countries in cooperation with national medical regulatory agencies, pharmaceutical companies, and international agencies, all of which have an urgent interest and investment in ensuring that patients throughout the world have access to good quality medicines. The organization would also advocate strongly for including targets for achieving good quality medicines in the United Nations Millennium Development Goals and Sustainable Development Goals.


title: Developing vaccines against epidemic-prone emerging infectious diseases

text: Today’s world is characterized by increasing population density, human mobility, urbanization, and climate and ecological change. This global dynamic has various effects, including the increased appearance of emerging infectious diseases (EIDs), which pose a growing threat to global health security. Outbreaks of EIDs, like the 2013–2016 Ebola outbreak in West Africa or the current Ebola outbreak in Democratic Republic of the Congo (DRC), have not only put populations in low- and middle-income countries (LMIC) at risk in terms of morbidity and mortality, but they also have had a significant impact on economic growth in affected regions and beyond. The Coalition for Epidemic Preparedness Innovation (CEPI) is an innovative global partnership between public, private, philanthropic, and civil society organizations that was launched as the result of a consensus that a coordinated, international, and intergovernmental plan was needed to develop and deploy new vaccines to prevent future epidemics. CEPI is focusing on supporting candidate vaccines against the World Health Organization (WHO) Blueprint priority pathogens MERS-CoV, Nipah virus, Lassa fever virus, and Rift Valley fever virus, as well as Chikungunya virus, which is on the WHO watch list. The current vaccine portfolio contains a wide variety of technologies, ranging across recombinant viral vectors, nucleic acids, and recombinant proteins. To support and accelerate vaccine development, CEPI will also support science projects related to the development of biological standards and assays, animal models, epidemiological studies, and diagnostics, as well as build capacities for future clinical trials in risk-prone contexts.


docid: uapxlnib

title: Evaluation of the synergetic effect of Yupingfeng san and Flos Sophorae Immaturus based on free radical scavenging capacity

text: OBJECTIVE: This study aimed to determine the optimal extraction process and examine whether the combination ofFlos Sophorae Immaturus (FSI) and Yupingfeng san (YPS) has a synergistic effect on free radical scavenging capacity. DESIGN AND METHODS: The time of immersion and extraction and the ratios (material/solvent) of the combination of YPS and FSI were optimized on the basis of polysaccharide and flavonoid yields via orthogonal design. The optimal result was used in the 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay and animal experiments to test the antioxidant activity, which is reflected by superoxide dismutase, malondialdehyde, glutathione peroxidase, and total antioxidant capacity serum levels. The optimal extraction process was determined using various ingredients to obtain complex extracts with high active ingredient content and antioxidant activity. DPPH assay results showed that the optimized ingredients have antioxidant effects, and the combination had better antioxidation function than YPS in vitro. The combination also showed synergistic antioxidant activity compared with YPS in vivo. CONCLUSIONS: The combination of YPS and FSI had a synergistic antioxidant effect in vitro. The optimized extracts had antioxidant effects in vivo. These results indicated that YPS could be used with FSI to improve its antioxidant capacity in the body on the basis of free radical scavenging capacity.


title: Regulation of Stress Responses and Translational Control by Coronavirus

text: Similar to other viruses, coronavirus infection triggers cellular stress responses in infected host cells. The close association of coronavirus replication with the endoplasmic reticulum (ER) results in the ER stress responses, which impose a challenge to the viruses. Viruses, in turn, have come up with various mechanisms to block or subvert these responses. One of the ER stress responses is inhibition of the global protein synthesis to reduce the amount of unfolded proteins inside the ER lumen. Viruses have evolved the capacity to overcome the protein translation shutoff to ensure viral protein production. Here, we review the strategies exploited by coronavirus to modulate cellular stress response pathways. The involvement of coronavirus-induced stress responses and translational control in viral pathogenesis will also be briefly discussed.


docid: wmelyonk

title: A proposed treatment for pathogenic enveloped viruses having high rates of mutation or replication

text: Several enveloped viruses, particularly some RNA viruses, have high rates of mutation or replication, which can make them virulent pathogens in humans and other mammals. A proposed treatment could use synthesized proteins to mask pathogenic viral surface proteins to quickly induce an immune attack on specific enveloped viruses by using existing immune cells. One treatment could inject dual‐protein ligand masks into patients' blood streams to mask pathogenic surface proteins used to infect mammalian cells. The mammalian immune system already uses an analogous, more complex structure called a pentraxin to neutralize some pathogens by connecting their surface proteins to immune cells. And several types of antiviral peptides have already experimentally demonstrated effectiveness in blocking various viral pathogen infections. These treatments offer advantages, especially for currently untreatable viral pathogens. Furthermore, using dual‐protein ligands and the antigenic memory of some subpopulations of NK cells would also allow the creation of defacto vaccines based on a host's NK cells, instead of vaccines utilizing CD4 and CD8 α:β T cells, which are limited by the requirement of MHC presentation of the target antigens to α:β T cells. Targeted NK cell vaccines could attack host cells latently or actively infected by intracellular pathogens, even host cells having pathogen downregulated MHC antigen presentation. Eight postulates concerning the effects of pathogen mutation, or change in phenotype from genetic recombination or rearrangement, and replication rates on pathogen versus host dominance are also listed, which should be applicable to viral and non‐viral pathogens.


docid: ehhotvqc

title: Stepwise prediction and statistical screening: B-cell epitopes on neuraminidase of human avian H(5)N(1) virus

text: The B-cell epitopes of virus are associated with the antiviral drug and the vaccine screening. As the nucleotide sequences of neuraminidase (NA) of stain GD-01-06 were sequenced, we predicted the α-helix and β-fold structure and the indexes of the flexible regions of secondary structure of NA with methods of the Hydrophilicity plot by Kyte-Doolittle, the Surface probability plot by Emini and the Antigenic index by Jameson-Wolf, and then screened statistically the parameters to predict B-cell epitopes by the Hierarchical cluster and the Bivariate correlation and the quartiles with SPSS 13.0. The impact of variation of amino acids in NA on its epitopes was analyzed. The predictive results were evaluated by Wu’s Antigenic Index and SWISS-MODEL. We found that the most possible epitopes on NA were located within or nearby its N-terminal Nos. 120–137, 81–84, 408–415, 273–282, 429–432, 356–368, 46–55, 146–155, 341–350 and 198–209, which were the dominant regions of NA epitopes. Peptide 120–137 including the glycoprotein domain (NGT(126–128)) was first chosen as the B-cell epitopes on NA. NA in H(5)N(1) strain isolated after 2003 lacked in No. 53 amino acid (I), resulting in an increase in the surface flexible region of NA in GD-01-06 and an enlargement to their epitope regions (VEP(46–48) → VEPISNTNFL(46–55)). Conclusively, prediction of the B-cell epitopes on the NA based on multiple parameters is useful for researches on the molecular immunology and drug screening and immuno-prophylaxis. A deletion of No. 53 amino acid (I) in NA in strain GD-01-06 might increase its antigenicity.