title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

text: Middle East respiratory syndrome coronavirus (MERS-CoV) infection is associated with a high case-fatality rate, and the potential pandemic spread of the virus is a public health concern. The spike protein of MERS-CoV (MERS-S) facilitates viral entry into host cells, which depends on activation of MERS-S by cellular proteases. Proteolytic activation of MERS-S during viral uptake into target cells has been demonstrated. However, it is unclear whether MERS-S is also cleaved during S protein synthesis in infected cells and whether cleavage is required for MERS-CoV infectivity. Here, we show that MERS-S is processed by proprotein convertases in MERS-S–transfected and MERS-CoV–infected cells and that several RXXR motifs located at the border between the surface and transmembrane subunit of MERS-S are required for efficient proteolysis. However, blockade of proprotein convertases did not impact MERS-S–dependent transduction of target cells expressing high amounts of the viral receptor, DPP4, and did not modulate MERS-CoV infectivity. These results show that MERS-S is a substrate for proprotein convertases and demonstrate that processing by these enzymes is dispensable for S protein activation. Efforts to inhibit MERS-CoV infection by targeting host cell proteases should therefore focus on enzymes that process MERS-S during viral uptake into target cells.


Hidden risk of nosocomial transmission: a presymptomatic novel coronavirus disease-19 (COVID-19) case with ischemic stroke


title: A Review of Laboratory-Acquired Infections in the Asia-Pacific: Understanding Risk and the Need for Improved Biosafety for Veterinary and Zoonotic Diseases

text: A rapid review was performed to determine (1) the number and causes of reported laboratory-acquired infections (LAI) in the Asia-Pacific region; (2) their significance and threat to the community; (3) the primary risk factors associated with LAIs; (4) the consequences in the event of a LAI or pathogen escape; and (5) to make general recommendations regarding biosafety practices for diagnosis and research in the Asia-Pacific region. A search for LAI and zoonoses in the Asia-Pacific region using online search engines revealed a relatively low number of reports. Only 27 LAI reports were published between 1982 and 2016. The most common pathogens associated with LAIs were dengue virus, Arthroderma spp., Brucella spp., Mycobacterium spp., Rickettsia spp., and Shigella spp. Seventy-eight percent (21 out of 27 LAI reports) occurred in high-income countries (i.e., Australia, Japan, South Korea, Singapore, and Taiwan) where laboratories were likely to comply with international biosafety standards. Two upper-middle income countries (China (2), and Malaysia (2)) and one lower-middle income country (India (2)) reported LAI incidents. The majority of the reports (fifty-two percent (14/27)) of LAIs occurred in research laboratories. Five LAI reports were from clinical or diagnostic laboratories that are considered at the frontier for zoonotic disease detection. Governments and laboratories in the Asia-Pacific region should be encouraged to report LAI cases as it provides a useful tool to monitor unintended release of zoonotic pathogens and to further improve laboratory biosafety. Non-reporting of LAI events could pose a risk of disease transmission from infected laboratory staff to communities and the environment. The international community has an important and continuing role to play in supporting laboratories in the Asia-Pacific region to ensure that they maintain the safe working environment for the staff and their families, and the wider community.


title: Covid‐19 and diabetes mellitus: unveiling the interaction of two pandemics

text: A novel RNA betacoronavirus causing coronavirus disease 2019 (Covid‐19) has now been declared pandemic disease by WHO. Guo et al published the first report of biochemical features in patients with diabetes and the further risk that this disease can determine to the progression of Covid‐19. Among different cytokines found significantly higher in patients with diabetes compared to those without, Interleukin‐6 (IL‐6), which is already increased in conditions of chronic inflammation, may play a more deleterious role in Covid‐19 infection. Targeting the overexpression of Il‐6 effects with a monoclonal antibody against IL‐6 receptor or using Janus Kinase inhibitors may be particularly helpful for treatment of Covid‐19 pneumonia in diabetes.


title: Clinical and laboratory profiles of hospitalized children with acute respiratory virus infection

text: PURPOSE: Despite the availability of molecular methods, identification of the causative virus in children with acute respiratory infections (ARIs) has proven difficult as the same viruses are often detected in asymptomatic children. METHODS: Multiplex reverse transcription polymerase chain reaction assays were performed to detect 15 common respiratory viruses in children under 15 years of age who were hospitalized with ARI between January 2013 and December 2015. Viral epidemiology and clinical profiles of single virus infections were evaluated. RESULTS: Of 3,505 patients, viruses were identified in 2,424 (69.1%), with the assay revealing a single virus in 1,747 cases (49.8%). While major pathogens in single virus-positive cases differed according to age, human rhinovirus (hRV) was common in patients of all ages. Respiratory syncytial virus (RSV), influenza virus (IF), and human metapneumovirus (hMPV) were found to be seasonal pathogens, appearing from fall through winter and spring, whereas hRV and adenovirus (AdV) were detected in every season. Patients with ARIs caused by RSV and hRV were frequently afebrile and more commonly had wheezing compared with patients with other viral ARIs. Neutrophil-dominant inflammation was observed in ARIs caused by IF, AdV, and hRV, whereas lymphocyte-dominant inflammation was observed with RSV A, parainfluenza virus, and hMPV. Monocytosis was common with RSV and AdV, whereas eosinophilia was observed with hRV. CONCLUSION: In combination with viral identification, recognition of virus-specific clinical and laboratory patterns will expand our understanding of the epidemiology of viral ARIs and help us to establish more efficient therapeutic and preventive strategies.


Le protocole d’accord revisité pour atténuer les pertes des LBM


Routine in an Italian high-volume Vascular Surgery Unit during the COVID-19 era: how the pandemic changed the vascular daily practice


title: Modulation of HIV-1 immunity by adjuvants

text: PURPOSE OF REVIEW: To summarize the role of adjuvants in eliciting desirable antibody responses against HIV-1 with particular emphasis on both historical context and recent developments. RECENT FINDINGS: Increased understanding of the role of pattern recognition receptors such as Toll-like receptors in recruiting and directing the immune system has increased the variety of adjuvant formulations being tested in animal models and humans. Across all vaccine platforms, adjuvant formulations have been shown to enhance desirable immune responses such as higher antibody titers and increased functional activity. Although no vaccine formulation has yet succeeded in eliciting broad neutralizing antibodies against HIV-1, the ability of adjuvants to direct the immune response to immunogens suggests they will be critically important in any successful HIV-1 vaccine. SUMMARY: The parallel development of adjuvants along with better HIV-1 immunogens will be needed for a successful AIDS vaccine. Additional comparative testing will be required to determine the optimal adjuvant and immunogen regimen that can elicit antibody responses capable of blocking HIV-1 transmission.


(839) Viral Pneumonia Accelerates Graft Death and CLAD in Lung Transplant Recipients


title: Study of expiratory droplet dispersion and transport using a new Eulerian modeling approach

text: Abstract Understanding of droplet nuclei dispersion and transport characteristics can provide more engineering strategies to control transmission of airborne diseases. Droplet dispersion in a room under the conventional well-mixed and displacement ventilation is simulated. Two droplet nuclei sizes, 0.01 and 10μm, are selected as they represent very fine and coarse droplets. The flow field is modeled using k–ε RNG model. A new Eulerian drift-flux methodology is employed to model droplet phase. Under the conventional ventilation scheme, both fine and coarse droplets are homogeneously dispersed within approximately 50s. Droplet nuclei exhibit distinctive dispersion behavior, particularly for low airflow microenvironment. After 270s of droplet emission, gravitational settling influences the dispersion for 10μm droplets, and concentration gradient can still be observed for displacement ventilation.


title: Risk for HIV-1 infection is not associated with Repeat-Region polymorphism in the DC-SIGN neck domain and Novel Genetic DC-SIGN Variants among North Indians

text: Abstract Background Several genetic factors have been related to HIV-1 resistance, the homozygosity for a mutation in CCR5 gene (CCR5Δ 32 allele) is presently considered the most relevant one. The C-type lectin, DC-SIGN efficiently binds and transmits HIV-1 to susceptible cell in trans thereby augmenting the infection. A potential association of the DC-SIGN neck domain repeats polymorphism and risk of HIV-1 infection is currently under debate. Methods Genetic risk association study was conducted in HIV-1 exposed seronegative (HES; n =50) individuals, HIV-1 seronegative (HSN; n =314) healthy control and HIV-1 infected seropositive patients (HSP; n =190) for polymorphism in neck domain of DC-SIGN gene. The DC-SIGN genotypes were identified by PCR from DNA extracted from peripheral blood and confirmed by sequencing. Fisher exact or χ2 test was used for statistical analysis. Results One HSN and HSP individual who were heterozygous (7/8) with respect to DC-SIGN repeat regions were found. The DC-SIGN neck repeat polymorphism among North Indian individuals was not associated with susceptibility to HIV-1 infection. Furthermore, inheritance study of heterozygous mutation (7/8) in HSN individual's family showed that one parent, two brothers, one sister and one daughter were heterozygous (7/8) for DC-SIGN mutant allele. Sequence analyses of DC-SIGN exon 4 repeat region of randomly selected 25 North Indian individuals from HSP, HSN and HES revealed four conserved intronic mutations. These mutations were at nucleotide position 1283, 1306, 1308 upstream and 1906 downstream of the DC-SIGN exon 4 repeat region when compared with the wild type sequence (NCBI Acc. No. AF209479). Conclusion The polymorphism in DC-SIGN neck repeats region was rare and not associated with HIV-1 susceptibility among North Indians. Sequencing analysis of DC-SIGN gene confirmed four novel genetic variants in intronic region flanking exon 4 coding region.


title: Research Advances in How the cGAS-STING Pathway Controls the Cellular Inflammatory Response

text: Double-stranded DNA (dsDNA) sensor cyclic-GMP-AMP synthase (cGAS) along with the downstream stimulator of interferon genes (STING) acting as essential immune-surveillance mediators have become hot topics of research. The intrinsic function of the cGAS-STING pathway facilitates type-I interferon (IFN) inflammatory signaling responses and other cellular processes such as autophagy, cell survival, senescence. cGAS-STING pathway interplays with other innate immune pathways, by which it participates in regulating infection, inflammatory disease, and cancer. The therapeutic approaches targeting this pathway show promise for future translation into clinical applications. Here, we present a review of the important previous works and recent advances regarding the cGAS-STING pathway, and provide a comprehensive understanding of the modulatory pattern of the cGAS-STING pathway under multifarious pathologic states.


title: Origin of measles virus: divergence from rinderpest virus between the 11(th )and 12(th )centuries

text: Measles, caused by measles virus (MeV), is a common infection in children. MeV is a member of the genus Morbillivirus and is most closely related to rinderpest virus (RPV), which is a pathogen of cattle. MeV is thought to have evolved in an environment where cattle and humans lived in close proximity. Understanding the evolutionary history of MeV could answer questions related to divergence times of MeV and RPV. We investigated divergence times using relaxed clock Bayesian phylogenetics. Our estimates reveal that MeV had an evolutionary rate of 6.0 - 6.5 × 10(-4 )substitutions/site/year. It was concluded that the divergence time of the most recent common ancestor of current MeV was the early 20(th )century. And, divergence between MeV and RPV occurred around the 11(th )to 12(th )centuries. The result was unexpected because emergence of MeV was previously considered to have occurred in the prehistoric age. MeV may have originated from virus of non-human species and caused emerging infectious diseases around the 11(th )to 12(th )centuries. In such cases, investigating measles would give important information about the course of emerging infectious diseases.


title: A new Schiff base coordinated copper(II) compound induces apoptosis and inhibits tumor growth in gastric cancer

text: BACKGROUND: Gastric cancer, as a multifactorial disorders, shows cytological and architectural heterogeneity compared to other gastrointestinal cancers, making it therapeutically challenging. Cisplatin is generally used in clinic for gastric cancer treatment but with toxic side effects and develops resistance. Anti-tumor properties of copper and its coordinated compounds have been explored intensively in recent years. METHODS: In this study, we synthesized a novel Schiff base copper coordinated compound (SBCCC) and examined its antitumor effects in two gastric cancer cell lines SGC-7901 and BGC-823 as well as a mouse model of gastric cancer. RESULTS: The results show that SBCCC can significantly inhibit the proliferation of gastric cancer cells in a dose- and time-dependent manner. The IC50 of SBCCC in SGC-7901 and BGC-823 cells is 1 μM, which is much less than cisplatin’s IC50. SBCCC induces apoptosis and causes cell cycle arrest at the G1 phase. SBCCC induces apoptosis via multiple pathways including inhibition of NF-κB, ROS production and autophagy. CONCLUSIONS: The synthesized SBCCC induced cancer cell death via inhibition of NF-κB, ROS production and autophagy. The multiple cell-killing mechanisms were important to overcome therapeutic failure because of multidrug-resistance of cancer cells. SBCCC, with a lower IC50 compared to cisplatin, could render it the potential to overcome the side-effect for clinical application.


title: Efficacy and safety of Echinaforce® in respiratory tract infections

text: Echinaforce® is the standardised extract of Echinacea purpurea from Bioforce, Switzerland. Recent studies show immunomodulation and broad antiviral effects against respiratory tract viruses. Haemagglutinin and Neuraminidase are blocked. In contrast to Oseltamivir no resistance is caused by Echinaforce®. A randomised, double-blind, placebo-controlled study over four months confirms that Echinaforce® supports the immune resistance and acts directly against a series of viruses. Echinaforce® is efficacious and safe in respiratory tract infections for long-term and short-term prevention as well as for acute treatment.


title: Universal Shelter-in-Place vs. Advanced Automated Contact Tracing and Targeted Isolation: A Case for 21st-Century Technologies for SARS-CoV-2 and Future Pandemics

text: Abstract Objective To model and compare effect of digital contact tracing versus shelter-in-place on SARS-CoV-2 spread. Methods Using a classical epidemiologic framework, and parameters estimated from literature published between February 1, 2020 and May 25, 2020, we modeled two non-pharmacologic interventions- shelter-in-place and digital contact tracing- to curb spread of SARS-CoV-2. For contact tracing, we assumed an advanced, automated contact tracing (AACT) application that sends alerts to individuals advising self-isolation based on individual exposure profile. Model parameters included percentage population ordered to shelter-in-place, adoption rate of AACT, and percentage individuals who appropriately follow recommendations. Under influence of these variables, number of individuals infected, exposed, and isolated were estimated. Results Without any intervention, a high rate of infection (>10 million) with early peak is predicted. Shelter-in-place results in rapid decline in infection rate at the expense of impacting a large population segment. The AACT model achieves reduction in infected and exposed individuals similar to shelter-in-place without impacting a large number of individuals. For example, a 50% AACT adoption rate mimics a shelter-in-place order for 40% of the population and results in >90% decrease in peak number of infections. However, as compared to shelter-in-place, with AACT significantly fewer individuals would be isolated. Conclusion Wide adoption of digital contact tracing can mitigate infection spread similar to universal shelter-in-place, but with considerably fewer individuals isolated.


title: East of nature. Accounting for the environments of social sciences

text: At the core of ecological economics is the image of the economy as an open system embedded in the natural environment whose carrying capacity is limited. The present paper revisits this image by drawing upon the constructivist implications of Luhmann's social systems theory. To Luhmann, the modern society consists of a multitude of social systems each bringing forth and observing their own environments. If the Luhmannian vision is accepted, then ecological economics can be said to privilege the observational perspective of natural sciences. The unfortunate consequence of this privileging is the underestimation of a broad range of multidimensional sustainability risks which are foregrounded by the numerous alternative observational perspectives which are just as legitimate. It is argued that, rather than relativizing the sustainability concerns of the modern ecological economics, the Luhmannian perspective generalizes and radicalizes them. In doing so, the latter perspective opens new possibilities not only for navigating these risks but also for envisioning new resources and solutions.


title: Pollen likely seasonal factor in inhibiting flu-like epidemics. A Dutch study into the inverse relation between pollen counts, hay fever and flu-like incidence 2016–2019

text: Abstract There is uncertainty if current models for the Covid-19 pandemic should already take into account seasonality. That is because current environmental factors do not provide a powerful explanation of such seasonality, especially given climate differences between countries with moderate climates. It is hypothesized that one major factor is overlooked: pollen count. Pollen are documented to invoke strong immune responses and might create an environmental factor that makes it more difficult for flu-like viruses to survive outside a host. This Dutch study confirms that there is a (highly) significant inverse correlation between pollen count and weekly changes in medical flu consults, and that there is a highly significant inverse correlation between hay fever incidence, as measured by prescribed medication revenues, and weekly flu consults. This supports the idea that pollen are a direct or indirect factor in the seasonality of flu-like epidemics. If seasonality will be observed during the covid-19 spread as well, it is not unlikely that pollen play a role.


title: Boceprevir, GC-376, and calpain inhibitors II, XII inhibit SARS-CoV-2 viral replication by targeting the viral main protease

text: A novel coronavirus SARS-CoV-2, also called novel coronavirus 2019 (nCoV-19), started to circulate among humans around December 2019, and it is now widespread as a global pandemic. The disease caused by SARS-CoV-2 virus is called COVID-19, which is highly contagious and has an overall mortality rate of 6.96% as of May 4, 2020. There is no vaccine or antiviral available for SARS-CoV-2. In this study, we report our discovery of inhibitors targeting the SARS-CoV-2 main protease (M(pro)). Using the FRET-based enzymatic assay, several inhibitors including boceprevir, GC-376, and calpain inhibitors II, and XII were identified to have potent activity with single-digit to submicromolar IC(50) values in the enzymatic assay. The mechanism of action of the hits was further characterized using enzyme kinetic studies, thermal shift binding assays, and native mass spectrometry. Significantly, four compounds (boceprevir, GC-376, calpain inhibitors II and XII) inhibit SARS-CoV-2 viral replication in cell culture with EC(50) values ranging from 0.49 to 3.37 μM. Notably, boceprevir, calpain inhibitors II and XII represent novel chemotypes that are distinct from known M(pro) inhibitors. A complex crystal structure of SARS-CoV-2 M(pro) with GC-376, determined at 2.15 Å resolution with three monomers per asymmetric unit, revealed two unique binding configurations, shedding light on the molecular interactions and protein conformational flexibility underlying substrate and inhibitor binding by M(pro). Overall, the compounds identified herein provide promising starting points for the further development of SARS-CoV-2 therapeutics.


COVID-19 Y LA NEUMOLOGÍA DEL SIGLO XXI: ¿RETO U OPORTUNIDAD?


title: Management of Patients on Dialysis and With Kidney Transplantation During the SARS-CoV-2 (COVID-19) Pandemic in Brescia, Italy

text: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also known as coronavirus disease (COVID-19), is a major pandemic challenging health care systems around the world. The optimal management of patients infected with COVID-19 is still unclear, although the consensus is moving toward the need of a biphasic approach. During the first phase of the disease (from onset of the symptoms up to 7–10 days) viral-induced effects are prominent, with the opportunity to institute antiviral therapy. In the second inflammatory phase of the disease, immunosuppressive strategies (for example with glucocorticoids or anticytokine drugs) may be considered. This latter stage is characterized by the development of progressive lung involvement with increasing oxygen requirements and occasionally signs of the hemophagocytic syndrome. The management of the disease in patients with kidney disease is even more challenging, especially in those who are immunosuppressed or with severe comorbidities. Here we present the therapeutic approach used in Brescia (Italy) for managing patients infected with COVID-19 who underwent kidney transplantation and are receiving hemodialysis. Furthermore, we provide some clinical and physiopathological background, as well as preliminary outcome data of our cohort, to better clarify the pathogenesis of the disease and clinical management.


title: COVID-19-Versorgung – Strategien der Taskforce Coronavirus und Erfahrungen von den ersten 115 Fällen am Universitätsklinikum Freiburg

text: Background The new pandemic coronavirus SARS-CoV-2 causing coronavirus disease-2019 (COVID-19) poses immense challenges to health care systems worldwide. In the current manuscript we summarize the strategies, organisational approaches and actions of the Task-force Coronavirus at the University Medical Center Freiburg. We also report on experiences with implementation of these approaches and treatment outcomes in the first 115 COVID patients. Methods Retrospective, narrative process description and analysis of the time period between end of January and beginning of April 2020, performed by representatives of the involved departments and institutes. Additionally a retrospective observational cohort study with descriptive analysis of epidemiological and clinical data of COVID patients admitted until March 31st was performed. Results A multidisciplinary Task-force Coronavirus initiated measures concerning outpatient testing and counseling, reorganisation and separation of patient flow processes alongside with substantial escalation of inpatient capacities on regular wards and intensive care units. Within the framework of the resulting dynamic care model, 115 patients suffering from COVID could be treated without shortages in staff or bed capacities. Dicussion In the upcoming pandemic, adequate COVID management and care could be secured by a collaborative approach with inclusion of administrative departments, clinical disciplines and theoretical institutes of the University Medical Center Freiburg.


Fears of “highly catastrophic” COVID-19 spread in Yemen


title: Integrating mental health into primary care: the policy maker’s perspective and experience in China

text: In China, ‘community’ was an alien word. Many people used to live in dormitories (Danwei), to which they were assigned by government according to their work units. ‘Dormitory form’ community was closely linked to where people worked, and thus administration and supervision were simple, as was the provision of health services. In each Danwei, a clinic provided basic healthcare not only for its employees but also for the other residents of the dormitory. The old primary care service was based on this. In fact, the ‘golden age’ of community mental healthcare was at that time, when psychiatric hospitals extended their service to communities via the Danwei ’s clinics in the cities and via ‘barefoot doctors’ in the rural areas. Home beds, occupational therapy stations and shelter factories were set up in some cities and mobile mental health teams played important roles in the villages. Although this did not really represent the ‘integration’ of mental health into primary care, it was a good example of maximising the utilisation of the very limited mental health resources by stretching the psychiatric service, using administrative power, and mobilising family members (Shen et al, 1990; Zhang & Yan, 1990; Zhang, 1999).


title: Respirator Donning in Post-Hurricane New Orleans

text: We evaluated correctness of N95 filtering facepiece respirator donning by the public in post-hurricane New Orleans, where respirators were recommended for mold remediation. We randomly selected, interviewed, and observed 538 participants, using multiple logistic regression for analysis. Only 129 (24%) participants demonstrated proper donning. Errors included nose clip not tightened (71%) and straps incorrectly placed (52%); 22% put on the respirator upside down. Factors independently associated with proper donning were as follows: ever having used a mask or respirator (odds ratio [OR] 5.28; 95% confidence interval [CI], 1.79–22.64); ever having had a respirator fit test (OR 4.40; 95% CI, 2.52–7.81); being male (OR 2.44; 95% CI, 1.50–4.03); Caucasian race (OR 2.09; 95% CI, 1.32–3.33); having a certified respirator (OR 1.99, 95% CI 1.20–3.28); and having participated in mold clean-up (OR 1.82; 95% CI,1.00–3.41). Interventions to improve respirator donning should be considered in planning for influenza epidemics and disasters.


title: Peer mentoring for medical students during COVID‐19 pandemic via a social media platform

text: In many contexts, medical students collaborate with health care workers to deliver patient management and care in emergencies like the COVID‐19 pandemic. In others, medical students are experiencing an unintended pause in their education due to global university closure over COVID‐19 concerns. In either situation, students find themselves coping with mental and emotional issues, including stress, anxiety, and fear, that may require significant psychological and physical effort. Therefore, it is important that medical schools not only care about students' mental health but also implement strategies to support their understanding of crisis management, self‐mental care, and other principal measures in order to strengthen their coping skills and mental preparedness.


Syncope, Near-syncope, or Non-mechanical Falls as a Presenting Feature of COVID-19


title: Early assessment of the impact of mitigation measures on the COVID-19 outbreak in Italy

text: Abstract Background On March 11, 2020, the World Health Organization characterized the novel coronavirus (COVID-19) outbreak as a pandemic. The first cases in Italy were reported on January 30, 2020, and the outbreak quickly escalated. On March 19, 2020, deaths in Italy surpassed those in China. The Italian government implemented progressively restrictive measures leading to a nationwide lockdown on March 8, 2020. This study aimed to assess the impact of mitigation measures implemented in Italy on the spread of COVID-19. Methods Publicly available data were used to evaluate changes in the growth curve of the number of patients hospitalized in intensive care (IC) at three time intervals between February 19, 2020 and April 9, 2020, following the implementation of progressive measures: (1) containment and travel restrictions, (2) lockdown of the epicenter of the outbreak, (3) school closures and nationwide lockdown. The models that showed the highest reliability according to the Akaike Information Criterion and based on data from the three time intervals were projected to assess how the epidemic would have evolved if no other measure had been implemented. Results The most reliable models were (1) exponential, (2) quadratic and (3) cubic (R2 0.99, > 0.99 and > 0.99 respectively), indicating a progressive decrease in the growth of the curve. Conclusion This study suggests the measures were effective in flattening the epidemic curve and bought valuable time, allowing for the number of IC beds to be nearly doubled before the national health system reached maximum capacity.


title: Evaluating the efficiency of specimen pooling for PCR‐based detection of COVID‐19

text: In the age of a pandemic, such as the ongoing one caused by SARS‐CoV‐2, the world faces a limited supply of tests, personal protective equipment, and factories and supply chains are struggling to meet the growing demands. This study aimed to evaluate the efficacy of specimen pooling for testing of SARS‐CoV‐2 virus, to determine whether costs and resource savings could be achieved without impacting the sensitivity of the testing. Ten previously tested nasopharyngeal and throat swab specimens by real‐time PCR, were pooled for testing, containing either one or two known positive specimens of varying viral concentrations. Specimen pooling did not affect the sensitivity of detecting SARS‐CoV‐2 when the PCR cycle threshold (Ct) of original specimen was lower than 35. In specimens with low viral load (Ct>35), 2 out of 15 pools (13.3%) were false negative. Pooling specimens to test for COVID‐19 infection in low prevalence (≤1%) areas or in low risk populations can dramatically decrease the resource burden on laboratory operations by up to 80%. This paves the way for large‐scale population screening, allowing for assured policy decisions by governmental bodies to ease lockdown restrictions in areas with a low incidence of infection, or with lower risk populations. This article is protected by copyright. All rights reserved.


title: Dietary plasma proteins, the intestinal immune system, and the barrier functions of the intestinal mucosa(1)

text: The intestinal mucosa contributes to homeostasis by preventing the entrance of biological and chemical agents across the epithelium that could alter the stability of the system. This protective function is especially important at the time of weaning, when animals are exposed to infectious agents and to numerous stresses such as the change of environment and diet. Diets supplemented with spray-dried plasma or plasma protein fractions have been shown to improve growth performance of farm animals and have been proposed as an alternative to antibiotics. In this review, we summarize our findings on the mechanism of action of dietary plasma proteins using a rat model of intestinal inflammation, based on the administration of Staphylococcus aureus enterotoxin B (SEB). Staphylococcal enterotoxin B activates the gut-associated lymphoid tissue (GALT), increasing T-lymphocytes in Peyer's patches and the number of activated T lymphocytes in mesenteric lymph nodes (organized GALT). In the lamina propria SEB increased cytotoxic T δγ and natural killer cell populations of the diffuse GALT. Staphyloccocal enterotoxin B significantly increased proinflammatory cytokines in Peyer's patches and mucosa. Plasma protein supplements modulated the mucosal immune response in organized and diffuse GALT, protecting GALT from possible excessive activation by the SEB challenge. These effects are accompanied by a reduction of proinflammatory cytokine production, supporting the view that changes in cytokine production mediate the effects of dietary plasma proteins during intestinal inflammation. The increase in mucosal permeability and intestinal secretion induced by SEB was associated with decreased expression of mucosal tight-junction and adherent-junction proteins. Both plasma and plasma protein fractions prevented the effects of SEB on intestinal permeability, thus reducing the exposure of the host to microbial and food antigens across the interstitial space. These findings indicate that dietary plasma proteins modulate functional and structural properties of the intestinal mucosa.


title: Alternative Waste Management Strategies

text: The health care industry and, more specifically, the perioperative setting have a 30-year history of acquiring a complete disposable habit. The fast-paced nature of perioperative nursing allows little time to contemplate the costs or impact of these practices; however, nurses are increasingly aware and concerned for their own safety and the safety of those who may be exposed to potential disease-causing materials. It seems to be the time to reconsider the benefits and liabilities of our practice habits. In the absence of definitive, conclusive, and compelling unbiased scientific evidence, we are directed by industry rather than by our own profession.


An American's perspective living through COVID-19 in Taiwan


docid: ynfelmiy

title: A case series of COVID‐19 patients with chronic hepatitis B virus infection

text: Previous studies reported that coronavirus disease 2019 (COVID‐19) was likely to result in liver injury. However, few studies reported the impacts of COVID‐19 on liver function in patients with chronic liver diseases. We aimed to describe a case series of COVID‐19 patients with chronic hepatitis B virus (HBV) infection. Confirmed hospitalized COVID‐19 patients from hospitals in 10 cities of Jiangsu province, China were retrospectively included between January 18, 2020, and February 26, 2020. Demographic information, epidemiologic data, clinical features and treatment data were extracted from medical records. Seven COVID‐19 patients with chronic HBV infection were included. Six (85.7%) patients were male. The patients aged from 33 years to 49 years. Two patients had HBV‐related cirrhosis. One patients (14.3%) were positive for serum hepatitis B virus e antigen. On admission, 1 (14.3%) patients had mildly elevated ALT level (>40 U/L) and 1 (14.3%) had elevated AST level (>40 U/L). The serum albumin level and platelet counts were decreased in 2 patients with HBV‐related liver cirrhosis. Three (42.9%) patients had elevated ALT level and 2 (28.6%) patients had elevated AST level in hospitalization. However, the peak ALT and AST level during hospitalization was 51 U/L and 44 U/L, respectively. As of February 29, 2020, all patients were discharged. No patient was admitted to the intensive care units or developed liver failure during hospitalization. The abnormalities of liver function are not uncommon on COVID‐19 patients with chronic HBV infection in our case series. However, no patient developed severe liver‐related complications during hospitalization. This article is protected by copyright. All rights reserved.


title: Coronavirus-Induced Demyelination Occurs in the Presence of Virus-Specific Cytotoxic T Cells

text: Abstract C57BI/6, but not BALB/c, mice infected with mouse hepatitis virus strain JHM (MHV-JHM) develop a late onset, symptomatic demyelinating encephalomyelitis. In this report, we characterized anti-viral cytotoxic T cells in the central nervous system and spleen during the acute and chronic stages of the MHV infection. The data show that C57BI/6 mice display a cytotoxic T cell (CTL) response to the surface (S) glycoprotein and this response can be demonstrated in lymphocytes isolated from the brains and spinal cords of mice both acutely and persistently infected with MHV-JHM. Thus, the anti-S CTL activity present in the central nervous system of chronically infected animals is not sufficient to prevent the demyelinating process. BALB/c mice have been shown previously to mount a CTL response against the nucleocapsid (N) protein (Stohlman et al., 1992). Since C57BI/6 mice do not mount a response to the N protein, the role of the N-specific response in preventing the late onset disease was assessed using B10.A(18R) mice, recombinant in the H-2 locus. These mice contain the d alleles of the D and L loci and exhibit a CTL response against the N protein. However, unlike the BALB/c mice, these animals develop the late onset symptomatic disease. These results suggest that the N-specific response is partially protective against the development of the demyelinating disease, but that additional factors are also likely to be involved.


title: Evaluation of eluents for the recovery of an enveloped virus from hands by whole-hand sampling

text: AIMS: The objective of this research is to evaluate eluents for recovery of an enveloped bacteriophage, Φ6, using whole-hand sampling. METHODS AND RESULTS: Virus was applied to the hands of volunteers and sampled by the glove juice method with 1.5% beef extract (BE), phosphate buffered saline (PBS), 0.01 and 0.1% Tween 80, tryptic soy broth (TSB), and 9% NaCl. Each volunteer underwent multiple rounds application and hand sampling. Mean log(10) virus loss across trials was 2.6 for BE, 2.8 for PBS, 2.4 for TSB, 3.8 for NaCl, 3.0 for 0.1% Tween 80, and 2.9 for 0.01% Tween 80. Within each volunteer, there was a decline in viral loss from the first to last trial. CONCLUSIONS: These eluents can recover Φ6 from hands with ~2-3 log(10) loss, comparable to recoveries previously reported for influenza. Protein and detergent-based eluents may have similar recoveries, but recovery may still vary across repeated sampling. SIGNIFICANCE AND IMPACT: Based on current work, protein-based eluents such as beef extract can maximize recovery of enveloped viruses during hand sampling, providing methods for evaluating survival and transmission of enveloped viruses on hands. Further exploration is needed of the effect of repeated sampling on recovery from whole-hand sampling.


title: ApoD, a glia‐derived apolipoprotein, is required for peripheral nerve functional integrity and a timely response to injury

text: Glial cells are a key element to the process of axonal regeneration, either promoting or inhibiting axonal growth. The study of glial derived factors induced by injury is important to understand the processes that allow or preclude regeneration, and can explain why the PNS has a remarkable ability to regenerate, while the CNS does not. In this work we focus on Apolipoprotein D (ApoD), a Lipocalin expressed by glial cells in the PNS and CNS. ApoD expression is strongly induced upon PNS injury, but its role has not been elucidated. Here we show that ApoD is required for: (1) the maintenance of peripheral nerve function and tissue homeostasis with age, and (2) an adequate and timely response to injury. We study crushed sciatic nerves at two ages using ApoD knock‐out and transgenic mice over‐expressing human ApoD. The lack of ApoD decreases motor nerve conduction velocity and the thickness of myelin sheath in intact nerves. Following injury, we analyze the functional recovery, the cellular processes, and the protein and mRNA expression profiles of a group of injury‐induced genes. ApoD helps to recover locomotor function after injury, promoting myelin clearance, and regulating the extent of angiogenesis and the number of macrophages recruited to the injury site. Axon regeneration and remyelination are delayed without ApoD and stimulated by excess ApoD. The mRNA and protein expression profiles reveal that ApoD is functionally connected in an age‐dependent manner to specific molecular programs triggered by injury. © 2010 Wiley‐Liss, Inc.


Neumonía por virus parainfluenza tipo 4 y púrpura trombótica trombocitopénica


title: Laboratory evaluation and interpretation of synovial fluid

text: Canine and feline joint disease can be a primary disorder limited to joints or a manifestation of multisystemic disease. Collection and analysis of joint fluid provides valuable information for the diagnosis, prognosis, and treatment of diseases that affect the joint space. The cytologic recognition of the cellular components and infectious agents in synovial fluid categorizes the cell response and differentiates inflammatory and noninflammatory joint disorders. This information is supported by the cell counts, protein content, mucin clot test, bacterial culture, and serologic tests for infectious or immune-mediated disease. These results are integrated with the clinical history, physical examination, radiographic findings, and ancillary test results to arrive at a diagnosis and treatment plan.


title: Pediatric Asthma Mortality and Hospitalization Trends Across Asia Pacific Relationship With Asthma Drug Utilization Patterns

text: BACKGROUND: The wide variability in prevalence of childhood asthma across Asia Pacific is well documented, but less is known about its trends in mortality and hospitalization. OBJECTIVES: To examine pediatric asthma mortality and hospitalization trends of selected countries across Asia Pacific, and also patterns of asthma drug utilization. MATERIALS AND METHODS: Mortality and population data were sourced from the World Health Organization's mortality database. Data on hospitalization were obtained by direct inquiry and from government and scientific publications. Drug use for asthma was expressed as a controller-to-reliever (C:R) ratio (ie, units of inhaled corticosteroids/units of short-acting β-agonists, sold in each country). Time-series regression analyses were used to examine temporal patterns and study association between deaths, hospitalizations, and drug use. RESULTS: Japan showed a decreasing trend in pediatric asthma mortality whereas an increase was observed in Thailand. Hospitalizations decreased in Australia and Singapore but increased in Taiwan, Republic of China. C:R ratios increased significantly across the countries. CONCLUSIONS: Mixed trends in pediatric asthma mortality and hospitalization rates were observed, which coincided with a uniform increase in C:R ratios. This may reflect importance of other aspects of asthma management besides pharmacotherapy.


title: Pathogen Reduction: A Precautionary Principle Paradigm

text: Although remarkable advances have been made in the prevention of the major transfusion-transmitted diseases, long intervals have transpired between the first recognition of transfusion risk and the implementation of a preventive strategy. For hepatitis B virus, that interval was 30 years; for non-A, non-B/hepatitis C virus, 15 years; and for human immunodeficiency virus, West Nile virus, Trypanosoma cruzi, and bacteria, 3, 4, 5, and 18 years, respectively. In our existing reactive approach, there is a fundamental and inevitable delay before we can react; and thus, infections are destined to occur. The continued emergence or reemergence of transfusion-transmitted infections calls for a new paradigm of preemptive pathogen reduction (PR). Two PR systems, psoralen/UV-A and riboflavin/UV-A, have shown efficacy and safety for platelets and plasma; and psoralen/UV-A technology has been successfully implemented for platelets in Europe. Pathogen reduction can eliminate or reduce the risk for any nucleic acid containing agent, including bacteria, and thus will be effective for all but prion diseases. It is possible to introduce PR for platelets and plasma now and to concentrate resources on developing PR for red cells. This will require an intellectual and financial commitment from the National Institutes of Health, the Food and Drug Administration, industry, and the blood bank establishment, just as occurred for nucleic acid testing (NAT) technology. This can be done if there is sufficient will to do it.


title: Membrane penetrating peptides greatly enhance baculovirus transduction efficiency into mammalian cells

text: Abstract The baculovirus group of insect viruses is widely used for foreign gene introduction into mammalian cells for gene expression and protein production; however, the efficiency of baculovirus entry into mammalian cells is in general still low. In this study, two recombinant baculoviruses were engineered and their ability to improve viral entry was examined: (1) cytoplasmic transduction peptide (CTP) was fused with baculovirus envelope protein, GP64, to produce a cytoplasmic membrane penetrating baculovirus (vE-CTP); and (2) the protein transduction domain (PTD) of HIV TAT protein was fused with the baculovirus capsid protein VP39 to form a nuclear membrane penetrating baculovirus (vE-PTD). Transduction experiments showed that both viruses had better transduction efficiency than vE, a control virus that only expresses EGFP in mammalian cells. Interestingly, vE-CTP and vE-PTD were also able to improve the transduction efficiency of a co-transduced baculovirus, resulting in higher levels of gene expression. Our results have described new routes to further enhance the development of baculovirus as a tool for gene delivery into mammalian cells.


title: Preparedness for ongoing Ebola virus infection: how to welcome it?

text: Abstract The problem of Ebola virus infection is the big global concern. Preparedness for ongoing Ebola virus infection is the topic that should be discussed. In fact, it is necessary to set up a biosecurity system to protect against the present Ebola outbreak. The medical personnel have to prepare for fighting the problem. The management of the present outbreak requires international collaboration and control of cross-border disease transmission is also the big challenge. The good case study is the Hajj scenario.


title: Reliability of self-sampling for accurate assessment of respiratory virus viral and immunologic kinetics

text: The SARS-CoV-2 pandemic demonstrates the need for accurate and convenient approaches to diagnose and therapeutically monitor respiratory viral infections. We demonstrated that self-sampling with foam swabs is well-tolerated and provides quantitative viral output concordant with flocked swabs. Using longitudinal home-based self-sampling, we demonstrate nasal cytokine levels correlate and cluster according to immune cell of origin. Periods of stable viral loads are followed by rapid elimination, which could be coupled with cytokine expansion and contraction using mathematical models. Nasal foam swab self-sampling at home provides a precise, mechanistic readout of respiratory virus shedding and local immune responses.


title: Circulation of other respiratory viruses and viral co-infection during the 2009 pandemic influenza

text: Abstract Coinciding with the pandemic wave of the influenza A(H1N1)pdm09 virus, other respiratory viruses have co-circulated in our area and were responsible for many acute respiratory infections and influenza-like illness (ILI). Apart from the pandemic virus that was responsible for most ILI cases, incidence rates of other viruses have varied among geographical areas. In general, human rhinovirus was the most frequent among individuals from the community, and respiratory syncytial virus among hospitalized patients. Detection rates of other respiratory viruses such as human metapneumovirus, adenovirus or parainfluenza viruses have been much lower. On the basis of an interference mechanism, human rhinovirus may contribute to modulate the pandemic wave, although available data are not conclusive to support this hypothesis. In contrast, the epidemic wave of respiratory syncytial virus during 2009–2010 was similar to previous seasons. Overall, incidence rates of respiratory viruses other than influenza did not change significantly during the pandemic season compared to other seasons. No association has been found between coinfection of pandemic influenza and other respiratory viruses with the prognosis of patients with influenza. The involvement of clinical virology laboratories in the etiological diagnosis of ILI cases has improved and has optimized diagnostic procedures.


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title: Emerging and re-emerging bacterial zoonoses in Nigeria: current preventive measures and future approaches to intervention

text: A characteristic of bacterial zoonoses, diseases caused by bacteria that can be transmitted to humans from animals, is a propensity to re-emerge. Several studies demonstrate their ongoing transmission in Nigeria, the most populous country in Africa. However, as local epidemiological data on bacterial zoonoses are inadequate the extent and impact of these infectious diseases is under-reported. Consequently, they are not a targeted priority of national public health policies. This limited recognition is despite indications of their possible roles in the widespread prevalence of non-malarial undifferentiated fever in Nigeria. While a number of animal reservoirs and arthropod vectors have been identified in the transmission routes of these diseases, an escalation of cases of undiagnosed febrile illness highlights the urgent need for a comprehensive assessment of other potential reservoirs, vectors and transmission cycles that may increase the local risk of infection with bacterial zoonoses. Animal health interventions have been proposed as a cost-effective strategy. Here, we present a broad overview of bacterial zoonotic infections of humans in Nigeria in the context of evolving epidemiological patterns. Further, we propose that facilitating the operation of a community-based One Health program is essential to providing the comprehensive epidemiological information that is required to improve prioritization of bacterial zoonoses. This would provide a driver for much needed investment in relevant public health interventions in Africa's most populous country.


title: Identification of M.tuberculosis-Specific Th1 Cells Expressing CD69 Generated in vivo in Pleural Fluid Cells from Patients with Tuberculous Pleurisy

text: Th1 cell-mediated immune responses at the site of active infection are important to restrict the growth of M.tuberculosis (MTB) and for the spontaneous resolution of patients with tuberculous pleurisy (TBP). In the present study, we found that without any stimulation, CD4(+) T cells in pleural fluid cells (PFCs) from patients with TBP expressed significantly higher levels of CD69 than PBMCs from patients with tuberculosis (TB) or healthy donors. CD4(+)CD69(+) T cells expressed T-bet and IL-12Rβ2. After stimulation with MTB-specific antigens, CD4(+)CD69(+) T cells expressed significantly higher levels of IFN-γ, IL-2 and TNF-α than CD4(+)CD69(−) T cells, demonstrating that CD4(+)CD69(+) T cells were MTB-specific Th1 cells. In addition, CD4(+)CD69(+) T cells were mostly polyfunctional Th1 cells that simultaneously produced IFN-γ, IL-2, TNF-α and displayed an effector or effector memory phenotype (CD45RA(−)CCR7(−)CD62L(−)CD27(−)). Moreover, the percentages of CD4(+)CD69(+) T cells were significantly and positively correlated with polyfunctional T cells. Interestingly, sorted CD4(+)CD69(+) but not CD4(+)CD69(−) fractions by flow cytometry produced IFN-γ, IL-2 and TNF-α that were significantly regulated by CD4(+)CD25(+) Treg cells. Taken together, based on the expression of CD69, we found a direct quantitative and qualitative method to detect and evaluate the in vivo generated MTB-specific polyfunctional CD4(+) T cells in PFCs from patients with TBP. This method can be used for the potential diagnosis and enrichment or isolation of MTB-specific Th1 cells in the investigations.


Clinical considerations for managing dermatology patients on systemic immunosuppressive and/or biologic therapy during COVID-19 pandemic


title: Pyridine N-oxide derivatives are inhibitory to the human SARS and feline infectious peritonitis coronavirus in cell culture

text: Objectives: Evaluation of a wide variety of pyridine N-oxide derivatives on their inhibitory activity against feline coronavirus (FIPV strain) and human SARS-CoV (Frankfurt strain-1) in cell culture. Methods: FIPV and SARS-CoV were exposed to confluent Crandel feline kidney (CRFK) and simian kidney (Vero) cell cultures in the presence of serial concentrations of the test compounds. The anti-cytopathic activity of the pyridine N-oxide derivatives was monitored by spectrophotometric analysis. Results and conclusions: A wide variety of pyridine N-oxide derivatives have been found to be inhibitory against feline coronavirus (FIPV strain) and human SARS-CoV (Frankfurt strain-1) in CRFK and simian kidney (Vero) cell cultures, respectively. The oxide part on the pyridine moiety proved indispensable for anti-coronavirus activity. The potency and virus specificity of the pyridine N-oxide derivatives varied depending the nature and specific location of substituents (i.e. alkyl, halogeno, nitro, etc.) on the different parts of the molecule. The most selective compounds were active in the higher microgram per litre range, being non-toxic at 50–100 mg/L. There was a poor structure-antiviral activity relationship (SAR) for the pyridine N-oxide derivatives against Fe-CoV and SARS-CoV. One of the most active and selective compounds was shown to inhibit Fe-CoV replication at the transcriptional level.


title: Coronavirus disease 2019 (COVID-19): Experiences and protocols from the Department of Prosthodontics at the Wuhan University

text: ABSTRACT Statement of problem The novel Coronavirus Disease 2019 (COVID-19) is a global pandemic, and many countries and regions are still currently in the midst of the outbreak. This pandemic has caused prosthodontics units to suspend their clinical and educational operations in academia. Purpose The purpose of this article was to review the experiences from the Department of Prosthodontics, Wuhan University School and Hospital of Stomatology (DP-WHUSHS) during the COVID-19 outbreak and the protocols DP-WHUSHS utilized to resume clinical activities after the outbreak. Material and methods The descriptive approach was used in this article to provide a chronological narrative of the experiences and protocols from the DP-WHUSHS during the COVID-19 outbreak and after the outbreak. Results During the COVID-19 outbreak period, clinical care was provided for patients with dental emergencies using enhanced Grade 2 or Garde 3 personal protective equipment (PPE). Teledentistry was used to provide care for patients with non-emergency needs. Online webinars and lectures were provided for the predoctoral students, residents, and dentists to minimize the interruption in their education and engage the dental community amid the pandemic. Various factors were considered before clinical activities resumed after the outbreak subsided. Additional resources were allocated for facility preparation and management and employee training. New infection control and clinical operation protocols were developed to minimize the healthcare-associated infection of airborne transmission diseases. The psychological health and mental wellness of the employees were emphasized. Distance or online education is still under rapid development to provide students and dentists opportunities to advance their knowledge amid the pandemic. Conclusions Within the limitation of this descriptive review, the following conclusions were drawn. Patient welfare and emergency needs should be considered amid the pandemic. Enhanced Grade 2 or Grade 3 PPE should be used during the outbreak. Multifactorial considerations for work resumption after the outbreak included facility preparation and management, training for employees, and clinical operation management. In-person psychological consultation and online mental wellness programs were available to employees to improve their mental wellness. Distance or online education was under rapid development to minimize the interruption in education for the students and to engage the dental community amid a pandemic.


title: Is it Ethically Appropriate to Continue Surgical Clinical Trials During the COVID-19 Pandemic? EDITED BY DR SARR

text: COVID-19 has greatly impacted surgical care and decision-making. The status of surgical clinical trials during this pandemic has not been addressed. We provide a framework and recommendations for the management of patients involved in surgical clinical trials.


title: Inclusion of Health in Impact Assessment: A Review of Current Practice in Sub-Saharan Africa

text: Natural resource extraction projects, including those in the mining sector, have various effects on human health and wellbeing, with communities in resource-rich areas in sub-Saharan Africa (SSA) being particularly vulnerable. While impact assessments (IA) can predict and mitigate negative effects, it is unclear whether and to what extent health aspects are included in current IA practice in SSA. For collecting IA reports, we contacted 569 mining projects and 35 ministries regulating the mining sector. The reports obtained were complemented by reports identified in prior research. The examination of the final sample of 44 IA reports revealed a heavy focus on environmental health determinants and included health outcomes were often limited to a few aspects, such as HIV, malaria and injuries. The miniscule yield of reports (1.6% of contacted projects) and the low response rate by the contacted mining companies (18%) might indicate a lack of transparency in the IA process of the mining sector in SSA. To address the shortcomings identified, policies regulating IA practice should strengthen the requirements for public disclosure of IA reports and promote a more comprehensive inclusion of health in IA, be it through stand-alone health impact assessment or more rigorous integration of health in other forms of IA.


title: The SARS Coronavirus S Glycoprotein Receptor Binding Domain: Fine Mapping and Functional Characterization

text: The entry of the SARS coronavirus (SCV) into cells is initiated by binding of its spike envelope glycoprotein (S) to a receptor, ACE2. We and others identified the receptor-binding domain (RBD) by using S fragments of various lengths but all including the amino acid residue 318 and two other potential glycosylation sites. To further characterize the role of glycosylation and identify residues important for its function as an interacting partner of ACE2, we have cloned, expressed and characterized various soluble fragments of S containing RBD, and mutated all potential glycosylation sites and 32 other residues. The shortest of these fragments still able to bind the receptor ACE2 did not include residue 318 (which is a potential glycosylation site), but started at residue 319, and has only two potential glycosylation sites (residues 330 and 357). Mutation of each of these sites to either alanine or glutamine, as well as mutation of residue 318 to alanine in longer fragments resulted in the same decrease of molecular weight (by approximately 3 kDa) suggesting that all glycosylation sites are functional. Simultaneous mutation of all glycosylation sites resulted in lack of expression suggesting that at least one glycosylation site (any of the three) is required for expression. Glycosylation did not affect binding to ACE2. Alanine scanning mutagenesis of the fragment S319–518 resulted in the identification of ten residues (K390, R426, D429, T431, I455, N473, F483, Q492, Y494, R495) that significantly reduced binding to ACE2, and one residue (D393) that appears to increase binding. Mutation of residue T431 reduced binding by about 2-fold, and mutation of the other eight residues – by more than 10-fold. Analysis of these data and the mapping of these mutations on the recently determined crystal structure of a fragment containing the RBD complexed to ACE2 (Li, F, Li, W, Farzan, M, and Harrison, S. C., submitted) suggested the existence of two hot spots on the S RBD surface, R426 and N473, which are likely to contribute significant portion of the binding energy. The finding that most of the mutations (23 out of 34 including glycosylation sites) do not affect the RBD binding function indicates possible mechanisms for evasion of immune responses.


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title: National policies for paediatric universal BCG vaccination were associated with decreased mortality due to COVID‐19

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title: Does hospital work constitute a risk factor for Helicobacter pylori infection?

text: Summary The aim of this study was to assess whether clinical work constitutes a risk factor for Helicobacter pylori infection among employees in hospitals. The prevalence of H. pylori infection was analysed in 249 individuals employed in a university teaching hospital according to three categories of hospital workers: (A) personnel from gastrointestinal endoscopy units (N=92); (B) personnel from other hospital units with direct patient contact (N=105); and (C) staff from laboratories and other units with no direct patient contact (N=52). Stool samples from each subject were examined with a validated enzyme-linked immunosorbent assay for the presence of H. pylori antigens. A questionnaire inquiring about sociodemographic and occupational characteristics was completed by each participant. The prevalence of H. pylori infection was 37.0% in group A, 35.2% in group B and 19.2% in group C (P<0.05). Among the different healthcare categories, nurses had a significant higher prevalence of H. pylori infection (P<0.01). No significant association was found between the length of employment or exposure to oral and faecal secretions, and H. pylori infection. Hospital work involving direct patient contact seems to constitute a major risk factor for H. pylori infection compared with hospital work not involving direct patient contact.


title: Recent Advances in the Laboratory Diagnosis of Equine Parasitic Diseases

text: This article reviews recent advances in laboratory diagnosis of equine parasitic diseases. Laboratory diagnosis of most equine parasitic diseases continues to rely on standard methods. Only laboratory diagnostic testing for EPM, cryptosporidiosis, and giardiasis are included, and the criteria for testing and interpretation of results for each new diagnostic method are explained.


title: Coronavirus disease (Covid-19): how does the exercise practice in active people with Type 1 Diabetes change? A preliminary survey

text: Abstract Aims Coronavirus disease (Covid-19) could lead persons with pre-existing medical conditions to severe respiratory infections. The Italian Government introduced quarantine to limit viral transmission. This measure could lead people with type 1 diabetes (PWT1D) to disrupt daily care routine including PA practice with difficulties in glycemia management. This study aims to explore PA level in PWT1D before and during quarantine and to describe variation in glycemia values. Methods An online survey investigating medical factors and the perceived and PA level in pre-established period before and after the introduction of quarantine was developed. Comparison between pre and post quarantine was assessed by Wilcoxon Signed Ranks test for continuous variables. Results A total of 154 subjects satisfied the eligibility criteria (54.5% males, 44.8±12.5 years). We found a decrease of PA level (Godin Scale Score 25±1.7vs38.6±1.7 points), steps number and minutes of exercise (respectively 12.606±5026vs4.760±3.145 and 66±4vs38±3) and an increase of glycemia values (142.1± 25.4 mg/dLvs150.8±29.4 mg/dL) Conclusions PWT1D reported a decrease in exercise and worst glycemia. Although PWT1D tried to remain active, their PA level was inadequate to prevent glycemia rising. The difficult to maintain a glycemic control could expose patients to diabetes complications and to an higher risk to counteract infections.


The Late Arrival of COVID-19 in Africa - Mitigating Pan-Continental Spread


title: Mitigating the air pollution effect? The remarkable decline in the pollution-mortality relationship in Hong Kong

text: Abstract Using transboundary pollution from mainland China as an instrument, we show that air pollution leads to higher cardio-respiratory mortality in Hong Kong. However, the air pollution effect has dramatically decreased over the past two decades: before 2003, a 10-unit increase in the Air Pollution Index could lead to a 3.1% increase in monthly cardio-respiratory mortality, but this effect has declined to 0.5% using recent data and is no longer statistically significant. Exploratory analyses suggest that a well-functioning medical system and immediate access to emergency services can help mitigate the contemporaneous effects of pollution on mortality.


title: 6-Thioguanine blocks SARS-CoV-2 replication by inhibition of PLpro protease activities

text: A recently emerged betacoronavirus, SARS-CoV-2, has led to a global health crisis that calls for the identification of effective therapeutics for COVID-19 disease. Coronavirus papain-like protease (PLpro) is an attractive drug target as it is essential for viral polyprotein cleavage and for deconjugation of ISG15, an antiviral ubiquitin-like protein. We show here that 6-Thioguanine (6-TG) inhibits SARS-CoV-2 PLpro-catalyzed viral polyprotein cleavage and ISG15 deconjugation in cells and inhibits replication of SARS-CoV-2 in Vero-E6 cells and Calu3 cells at submicromolar levels. As a well-characterized FDA-approved orally delivered drug, 6-TG represents a promising therapeutic for COVID-19 and other emerging coronaviruses. One Sentence Summary A repurposed drug that targets an essential enzymatic activity of SARS-CoV-2 represents a promising COVID-19 therapeutic.


title: Crimean-Congo Hemorrhagic Fever in Humanized Mice Reveals Glial Cells as Primary Targets of Neurological Infection

text: Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral hemorrhagic disease seen exclusively in humans. Central nervous system (CNS) infection and neurological involvement have also been reported in CCHF. In the current study, we inoculated NSG-SGM3 mice engrafted with human hematopoietic CD34(+) stem cells with low-passage CCHF virus strains isolated from human patients. In humanized mice, lethal disease develops, characterized by histopathological change in the liver and brain. To date, targets of neurological infection and disease have not been investigated in CCHF. CNS disease in humanized mice was characterized by gliosis, meningitis, and meningoencephalitis, and glial cells were identified as principal targets of infection. Humanized mice represent a novel lethal model for studies of CCHF countermeasures, and CCHF-associated CNS disease. Our data suggest a role for astrocyte dysfunction in neurological disease and identify key regions of infection in the CNS for future investigations of CCHF.


Technological Innovation in Outpatient Assistance for Chronic Liver Disease and Liver Transplant Patients During the Coronavirus Disease Outbreak: A Method to Minimize Transmission


title: Characterization of a novel human calicivirus that may be a naturally occurring recombinant

text: We identified a Norwalk-like calicivirus (CV) whose genome likely was derived from naturally occurring recombination. This strain (Arg320) was detected by the EIA developed against recombinant Mexico virus (rMxV) capsids, but the viral RNA polymerase sequence was closer to Lordsdale virus, in a separate genetic cluster of Norwalk-like viruses. A 3.3 kb cDNA from the RNA polymerase region to the 3′ end of the genome of Arg320 was cloned and sequenced. The sequence demonstrated that the capsid region of Arg320 shared 95% amino acid identity with MxV, but 68% identity with Lordsdale virus, while the RNA polymerase region shared 95% identity with Lordsdale virus, but 87% identity with MxV. Pair-wise sequence comparisons identified a potential recombination site at the polymerase/capsid junction. This is the first example of a naturally occurring recombinant in the CV family. Further studies to search for and characterize other strains may be necessary for understanding the genetic diversity of the family.


title: A pilot study on the disinfection efficacy of localized UV on the flushing-generated spread of pathogens

text: Abstract The process of toilet-flushing can generate flushing-associated water droplets which can potentially expose humans to pathogen-laden aerosols. Very little is known about such aerosol dissemination or the means for minimizing exposure to these aerosols. This study has evaluated the efficacy of ultraviolet waveband C (UV-C) for disinfection of flushing-generated pathogen-laden aerosols through tests with localized disinfection systems for airborne and surface contaminations. Three types of bacteria were chosen for investigation: Staphylococcus epidermidis, Escherichia coli, and Salmonella typhimurium. Tests were conducted with UV-C tubes of 5 W and 10 W. High levels of disinfection efficacies were observed, ranging from 76% to 97% for bacteria-laden aerosols at sources of emission, and efficiencies of 53% to 79% for surface samples in localized systems. The results from the localized systems were further compared with those obtained with an upper-room ultraviolet germicidal irradiation (UVGI) system. As it is important to note, the UV-C doses and ozone emissions for the localized systems were found well below the limits recommended in current guidelines. This research has shown that the disinfection of flushing-generated pathogen-laden aerosols in proximity to the source of emission was more effective than at the more distant sites where aerosols may be dispersed to the environment.


General practitioner management of notifiable diseases is central to communicable disease control


Australia's national COVID‐19 primary care response


title: Targeting the SARS-CoV-2 Main Protease to Repurpose Drugs for COVID-19

text: Guided by a computational docking analysis, about 30 FDA/EMA-approved small molecule medicines were characterized on their inhibition of the SARS-CoV-2 main protease (M(Pro)). Of these tested small molecule medicines, six displayed an IC(50) value in inhibiting M(Pro) below 100 μM. Three medicines pimozide, ebastine, and bepridil are basic small molecules that are expected to exert a similar effect as hydroxychloroquine in raising endosomal pH for slowing down the SARS-CoV-2 entry into human cell hosts. Bepridil has been previously explored in a high dose as 100 mg/kg for treating diseases. Its high dose use will likely achieve dual functions in treating COVID-19 by both raising the endosomal pH to slow viral entry and inhibiting M(Pro) in infected cells. Therefore, the current study urges serious considerations of using bepridil in COVID-19 clinical tests.


title: A new look at an old virus: patterns of mutation accumulation in the human H1N1 influenza virus since 1918

text: BACKGROUND: The H1N1 influenza A virus has been circulating in the human population for over 95 years, first manifesting itself in the pandemic of 1917–1918. Initial mortality was extremely high, but dropped exponentially over time. Influenza viruses have high mutation rates, and H1N1 has undergone significant genetic changes since 1918. The exact nature of H1N1 mutation accumulation over time has not been fully explored. METHODS: We have made a comprehensive historical analysis of mutational changes within H1N1 by examining over 4100 fully-sequenced H1N1 genomes. This has allowed us to examine the genetic changes arising within H1N1 from 1918 to the present. RESULTS: We document multiple extinction events, including the previously known extinction of the human H1N1 lineage in the 1950s, and an apparent second extinction of the human H1N1 lineage in 2009. These extinctions appear to be due to a continuous accumulation of mutations. At the time of its disappearance in 2009, the human H1N1 lineage had accumulated over 1400 point mutations (more than 10% of the genome), including approximately 330 non-synonymous changes (7.4% of all codons). The accumulation of both point mutations and non-synonymous amino acid changes occurred at constant rates (μ = 14.4 and 2.4 new mutations/year, respectively), and mutations accumulated uniformly across the entire influenza genome. We observed a continuous erosion over time of codon-specificity in H1N1, including a shift away from host (human, swine, and bird [duck]) codon preference patterns. CONCLUSIONS: While there have been numerous adaptations within the H1N1 genome, most of the genetic changes we document here appear to be non-adaptive, and much of the change appears to be degenerative. We suggest H1N1 has been undergoing natural genetic attenuation, and that significant attenuation may even occur during a single pandemic. This process may play a role in natural pandemic cessation and has apparently contributed to the exponential decline in mortality rates over time, as seen in all major human influenza strains. These findings may be relevant to the development of strategies for managing influenza pandemics and strain evolution.


title: Application of Humanized Mice in Immunological Research

text: During the past decade, the development of humanized mouse models and their general applications in biomedical research greatly accelerated the translation of outcomes obtained from basic research into potential diagnostic and therapeutic strategies in clinic. In this chapter, we firstly present an overview on the history and current progress of diverse humanized mouse models and then focus on those equipped with reconstituted human immune system. The update advancement in the establishment of humanized immune system mice and their applications in the studies of the development of human immune system and the pathogenesis of multiple human immune-related diseases are intensively reviewed here, while the shortcoming and perspective of these potent tools are discussed as well. As a valuable bridge across the gap between bench work and clinical trial, progressive humanized mouse models will undoubtedly continue to play an indispensable role in the wide area of biomedical research.


title: An unusual case of bullous hemorrhagic vasculitis in a COVID‐19 patient

text: A novel Coronavirus strain, named “Severe Acute Respiratory Syndrome Coronavirus 2” (SARS‐CoV‐2) was recently identified as the etiological agent of the COronaVIrus Disease 2019 (COVID‐19). Interestingly, a consistent number of COVID‐19‐associated skin manifestations seem to share a certain degree of vascular damage as common pathogenetic mechanism(1). Vascular injury may be due to the direct damage of endothelial cells by the virus or may represents an epiphenomenon of a dysregulated host inflammatory responses triggered by the infection(2). Here, we describe an unprecedented case of leukocytoclastic vasculitis presenting with a hemorrhagic bullous eruption in a patient affected by COVID‐19.


title: New Tetromycin Derivatives with Anti-Trypanosomal and Protease Inhibitory Activities

text: Four new tetromycin derivatives, tetromycins 1–4 and a previously known one, tetromycin B (5) were isolated from Streptomyces axinellae Pol001(T) cultivated from the Mediterranean sponge Axinella polypoides. Structures were assigned using extensive 1D and 2D NMR spectroscopy as well as HRESIMS analysis. The compounds were tested for antiparasitic activities against Leishmania major and Trypanosoma brucei, and for protease inhibition against several cysteine proteases such as falcipain, rhodesain, cathepsin L, cathepsin B, and viral proteases SARS-CoV M(pro), and PL(pro). The compounds showed antiparasitic activities against T. brucei and time-dependent inhibition of cathepsin L-like proteases with K(i) values in the low micromolar range.


Seroprevalence of Middle East respiratory syndrome coronavirus (MERS-CoV) in public health workers responding to a MERS outbreak in Seoul, Republic of Korea, in 2015


title: Neumomediastino espontáneo: estudio descriptivo de nuestra experiencia basada en 36 casos

text: El neumomediastino espontáneo se define como la presencia de aire o gas en el mediastino de forma primaria. Presentamos todos los casos de neumomediastino espontáneo hallados en nuestro hospital entre enero de 1996 y diciembre de 2004. Para ello, elaboramos un protocolo de recogida de datos que incluyó: antecedentes personales, desencadenantes, semiología, radiología, tratamiento, estancia y complicaciones. En este período diagnosticamos 36 casos —25 varones (69,4%) y 11 mujeres (27,5%)—, con una edad media de 36,8 años (rango: 11-90) y estancia media de 8,56 días (rango: 1-53). El cuadro clínico más habitual fue el dolor torácico, aislado (27%) o asociado a disnea (19,4%). En 14 pacientes (38,8%) hubo factor desencadenante. No hubo morbimortalidad asociada al proceso. Por todo ello, se concluye que el neumomediastino espontáneo es una entidad infrecuente, con gran variabilidad clínica, cuyo diagnóstico correcto exige un alto índice de sospecha, y la radiografía de tórax es la prueba idónea para ello. Spontaneous pneumomediastinum is defined as a primary process characterized by the presence of air or gas in the mediastinum. We report all the cases of spontaneous pneumomediastinum diagnosed in our hospital between January 1996 and December 2004. We developed a protocol for data collection that included the following: medical history, triggers, signs, radiology, treatment, hospital stay, and complications. During this period we diagnosed 36 cases—25 men (69.4%) and 11 women (27.5%)—with a mean age of 36.8 years (range, 11-90 years) and a mean hospital stay of 8.56 days (range, 1-53 days). The most common clinical presentation was chest pain, either isolated (27%) or with associated dyspnea (19.4%). A triggering factor was identified for 14 patients (38.8%). There was no associated morbidity or mortality. In view of our findings, we concluded that spontaneous pneumomediastinum is an uncommon entity with considerable clinical variability and that correct diagnosis requires a high level of suspicion. Radiography provides the best evidence for diagnosis.


title: Identification of two mutation sites in spike and envelope proteins mediating optimal cellular infection of porcine epidemic diarrhea virus from different pathways

text: Entry of the α-coronavirus porcine epidemic diarrhea virus (PEDV) requires specific proteases to activate spike (S) protein for the membrane fusion of the virion to the host cell following receptor binding. Herein, PEDV isolate 85-7 could proliferate and induce cell–cell fusion in a trypsin independent manner on Vero cells, and eight homologous mutation strains were screened by continuous proliferation in the absence of trypsin on Vero cells. According to the whole genome sequence comparative analysis, we identified four major variations located in nonstructural protein 2, S, open reading frame 3, and envelope (E) genes, respectively. Comparative analyses of their genomic variations and proliferation characteristics identified a single mutation within the S2′ cleavage site between C30 and C40 mutants: the substitution of conserved arginine (R) by a glycine (G) (R895G). This change resulted in weaker cell–cell fusion, smaller plaque morphology, higher virus titer and serious microfilament condensation. Further analysis confirmed that this mutation was responsible for optimal cell-adaptation, but not the determinant for trypsin-dependent entry of PEDV. Otherwise, a novel variation (16–20 aa deletion and an L25P mutation) in the transmembrane domain of the E protein affected multiple infection processes, including up-regulation of the production of the ER stress indicator GRP78, improving the expression of pro-inflammatory cytokines IL-6 and IL-8, and promoting apoptosis. The results of this study provide a better understanding of the potential mechanisms of viral functional proteins in PEDV replication, infection, and fitness. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13567-017-0449-y) contains supplementary material, which is available to authorized users.


docid: giakcaki

title: A simpler and more cost-effective peptide biosynthetic method using the truncated GST as carrier for epitope mapping

text: There is a need to develop better methods for epitope mapping and/or identification of antibody-recognizing motifs. Here, we describe improved biosynthetic peptide (BSP) method using a newly developed plasmid pXXGST-3 as vector, which has a viral E7 gene in the cloning sites of pXXGST-1. It is crucial to employ pXXGST-3 instead of pXXGST-1, since it makes use of the BSP method simpler and easier to perform, and more cost-effective for epitope mapping. These merits are embodied in two aspects: i) convenient recovery of double enzyme-digested product due to the existence of 315 bp inserted between BamH I and Sal I sites, and thus greatly reducing the production of self-ligation clones, and ii) no longer requiring control protein when screening recombinant (r-) clones expressing 8/18mer peptides by running polyacrylamide gel electrophoresis. The protocol involves the following core steps: (i) design of plus and minus strands of DNA fragments encoding overlapping 8/18mer peptides; (ii) chemical synthesis of the designed DNA fragments; (iii) development of r-clones using pXXGST-3 vector expressing each 8/18mer peptide fused with truncated GST188 protein; (iv) screening r-clones by running the cell pellets from each induced clone on SDS-PAGE gel followed by sequencing of inserted DNA fragments for each verified r-clone; and (v) Western blotting with either monoclonal antibodies or polyclonal antibodies. This improved GST188-BSP method provides a powerful alternative tool for epitope mapping.


title: Infectious causes of multiple sclerosis

text: Summary Multiple sclerosis (MS) is a serious chronic neurological disorder in which demyelination and inflammation occur in the white matter of the CNS. The findings of many epidemiological studies and a discordance of MS in monozygotic twins suggest that the disorder is acquired. The most likely cause is a virus because more than 90% of patients with MS have high concentrations of IgG, manifest as oligoclonal bands, in the brain and CSF. Most chronic inflammatory CNS disorders are infectious. More indirect evidence that MS is caused by a virus is the association of several viruses with demyelinating encephalomyelitis in human beings, and the induction of demyelination in animals infected with viruses in research. Nevertheless, no virus has been isolated from the brains of patients who had MS. Molecular analysis of IgG gene specificity in the brain and CSF of those with MS has shown features of an antigen-driven response: clonal amplification and extensive somatic mutations. A viral antigen against which the IgG in MS brain and CSF is directed might be identified.


title: Foot-and-mouth disease virus induces lysosomal degradation of host protein kinase PKR by 3C proteinase to facilitate virus replication

text: Abstract The interferon-induced double-strand RNA activated protein kinase (PKR) plays important roles in host defense against viral infection. Here we demonstrate the significant antiviral role of PKR against foot-and-mouth disease virus (FMDV) and report that FMDV infection inhibits PKR expression and activation in porcine kidney (PK-15) cells. The viral nonstructural protein 3C proteinase (3Cpro) is identified to be responsible for this inhibition. However, it is independent of the well-known proteinase activity of 3Cpro or 3Cpro-induced shutoff of host protein synthesis. We show that 3Cpro induces PKR degradation by lysosomal pathway and no interaction is determined between 3Cpro and PKR. Together, our results indicate that PKR acts an important antiviral factor during FMDV infection, and FMDV has evolved a strategy to overcome PKR-mediated antiviral role by downregulation of PKR protein.


title: Peptide presentation by bat MHC class I provides new insight into the antiviral immunity of bats

text: Bats harbor many zoonotic viruses, including highly pathogenic viruses of humans and other mammals, but they are typically asymptomatic in bats. To further understand the antiviral immunity of bats, we screened and identified a series of bat major histocompatibility complex (MHC) I Ptal-N*01:01–binding peptides derived from four different bat-borne viruses, i.e., Hendra virus (HeV), Ebola virus (EBOV), Middle East respiratory syndrome coronavirus (MERS-CoV), and H17N10 influenza-like virus. The structures of Ptal-N*01:01 display unusual peptide presentation features in that the bat-specific 3–amino acid (aa) insertion enables the tight “surface anchoring” of the P1-Asp in pocket A of bat MHC I. As the classical primary anchoring positions, the B and F pockets of Ptal-N*01:01 also show unconventional conformations, which contribute to unusual peptide motifs and distinct peptide presentation. Notably, the features of bat MHC I may be shared by MHC I from various marsupials. Our study sheds light on bat adaptive immunity and may benefit future vaccine development against bat-borne viruses of high impact on humans.


title: Chapter Six Feline Coronaviruses Pathogenesis of Feline Infectious Peritonitis

text: Abstract Feline infectious peritonitis (FIP) belongs to the few animal virus diseases in which, in the course of a generally harmless persistent infection, a virus acquires a small number of mutations that fundamentally change its pathogenicity, invariably resulting in a fatal outcome. The causative agent of this deadly disease, feline infectious peritonitis virus (FIPV), arises from feline enteric coronavirus (FECV). The review summarizes our current knowledge of the genome and proteome of feline coronaviruses (FCoVs), focusing on the viral surface (spike) protein S and the five accessory proteins. We also review the current classification of FCoVs into distinct serotypes and biotypes, cellular receptors of FCoVs and their presumed role in viral virulence, and discuss other aspects of FIPV-induced pathogenesis. Our current knowledge of genetic differences between FECVs and FIPVs has been mainly based on comparative sequence analyses that revealed “discriminatory” mutations that are present in FIPVs but not in FECVs. Most of these mutations result in amino acid substitutions in the S protein and these may have a critical role in the switch from FECV to FIPV. In most cases, the precise roles of these mutations in the molecular pathogenesis of FIP have not been tested experimentally in the natural host, mainly due to the lack of suitable experimental tools including genetically engineered virus mutants. We discuss the recent progress in the development of FCoV reverse genetics systems suitable to generate recombinant field viruses containing appropriate mutations for in vivo studies.


title: Molecular detection of infectious bronchitis and avian metapneumoviruses in Oman backyard poultry

text: Abstract Infectious bronchitis virus (IBV) and avian metapneumovirus (aMPV) are economically important viral pathogens infecting chickens globally. Identification of endemic IBV and aMPV strains promotes better control of both diseases and prevents production losses. Orophrayngeal swab samples were taken from 2317 birds within 243 different backyard flocks in Oman. Swabs from each flock were examined by RT-PCR using part-S1 and G gene primers for IBV and aMPV respectively. Thirty-nine chicken flocks were positive for IBV. Thirty two of these were genotyped and they were closely related to 793/B, M41, D274, IS/1494/06 and IS/885/00. 793/B-like IBV was also found in one turkey and one duck flock. Five flocks were positive for aMPV subtype B. Though no disease was witnessed at the time of sampling, identified viruses including variant IBV strains, may still pose a threat for both backyard and commercial poultry in Oman.


title: Lack of adverse effects during a target animal safety trial of extended-release buprenorphine in Fischer 344 rats

text: Extended-release buprenorphine is an effective analgesic in laboratory animals, and its safety has been established in mice but not in rats. The authors used a target animal safety trial to evaluate the safety of extended-release buprenorphine in rats. Fischer 344 rats received post-surgical subcutaneous injections of 1.3 mg, 3.9 mg or 6.5 mg buprenorphine per kg body weight (two times, six times or ten times the intended dose, respectively), and their body weight, clinical signs and symptoms, clinical pathology and histopathology were monitored for 4 d. Body weight was not significantly different in rats that received buprenorphine compared with control rats. Signs of nausea-related behavior were observed in 25% of the rats treated with buprenorphine. Clinical pathology results for all rats were normal, and gross and microscopic histopathology examinations identified no substantial abnormalities, suggesting that this behavior was of minor consequence. Other adverse events previously reported to occur with opiate therapy, including weight loss and dermal lesions at drug injection sites, were not observed in this study. The results of this study show that post-surgical administration of an extended-release buprenorphine product is safe in Fischer 344 rats and does not necessarily cause substantial adverse effects, confirming that opiate therapy is a viable choice in laboratory animal medicine.


Foreword: Sexually Transmitted Diseases in the Midst of the Coronavirus Pandemic


docid: ueqxxzsl

title: Folding of the rabbit hemorrhagic disease virus capsid protein and delineation of N-terminal domains dispensable for assembly

text: Rabbit hemorrhagic disease virus (RHDV) and European brown hare syndrome virus (EBHSV) are caliciviruses that produce severe symptoms and are lethal to rabbits and hares. The folding of the capsid protein was studied by determination of the antigenic pattern of chimeric capsid proteins, composed of regions from RHDV and EBHSV capsid proteins. The anti-RHDV monoclonal antibody (MAb) E3, which is known to bind an external conformational epitope, recognized the RHDV C-terminal region. The anti-RHDV MAb A47, which binds a buried epitope, recognized the RHDV N-terminal region. Using a pGEX expression library, we more precisely mapped the MAb A47 epitope on a 31 residues length peptide, between residue 129 and 160 of the VP60, confirming its location in the N-terminal part of the protein. These results demonstrate that the C-terminal part of the protein is accessible to the exterior whereas the N-terminal domain of the protein constitutes the internal shell domain of the particle. With the aim of using virus-like particles (VLPs) of RHDV as epitope carriers or DNA transfer vectors, we produced in the baculovirus system three proteins, ΔN1, ΔN2 and ΔN3, truncated at the N terminus. The ΔN1 protein assembled into VLPs, demonstrating that the first 42 amino acid residues are not essential for capsid assembly. In contrast, ΔN2, from which the first 75 residues were missing, was unable to form VLPs. The small particles obtained with the ΔN3 protein lacking residues 31 to 93, located in the immunodominant region of the RHDV capsid protein, indicate that up to 62 amino acid residues can be eliminated without preventing assembly.


title: A single-dose ChAdOx1-vectored vaccine provides complete protection against Nipah Bangladesh and Malaysia in Syrian golden hamsters

text: Nipah virus (NiV) is a highly pathogenic re-emerging virus that causes outbreaks in South East Asia. Currently, no approved and licensed vaccine or antivirals exist. Here, we investigated the efficacy of ChAdOx1 NiV(B), a simian adenovirus-based vaccine encoding NiV glycoprotein (G) Bangladesh, in Syrian hamsters. Prime-only as well as prime-boost vaccination resulted in uniform protection against a lethal challenge with NiV Bangladesh: all animals survived challenge and we were unable to find infectious virus either in oral swabs, lung or brain tissue. Furthermore, no pathological lung damage was observed. A single-dose of ChAdOx1 NiV(B) also prevented disease and lethality from heterologous challenge with NiV Malaysia. While we were unable to detect infectious virus in swabs or tissue of animals challenged with the heterologous strain, a very limited amount of viral RNA could be found in lung tissue by in situ hybridization. A single dose of ChAdOx1 NiV(B) also provided partial protection against Hendra virus and passive transfer of antibodies elicited by ChAdOx1 NiV(B) vaccination partially protected Syrian hamsters against NiV Bangladesh. From these data, we conclude that ChAdOx1 NiV(B) is a suitable candidate for further NiV vaccine pre-clinical development.


THE COVID-19 HOTEL FOR HEALTHCARE WORKERS: AN ITALIAN BEST PRACTICE


title: Hyperinflammation and derangement of renin-angiotensin-aldosterone system in COVID-19: A novel hypothesis for clinically suspected hypercoagulopathy and microvascular immunothrombosis

text: Abstract Early clinical evidence suggests that severe cases of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are frequently characterized by hyperinflammation, imbalance of renin-angiotensin-aldosterone system, and a particular form of vasculopathy, thrombotic microangiopathy, and intravascular coagulopathy. In this paper, we present an immunothrombosis model of COVID-19. We discuss the underlying pathogenesis and the interaction between multiple systems, resulting in propagation of immunothrombosis, which through investigation in the coming weeks, may lead to both an improved understanding of COVID-19 pathophysiology and identification of innovative and efficient therapeutic targets to reverse the otherwise unfavorable clinical outcome of many of these patients.


Sport in the tracks and fields of the corona virus: critical issues during the exit from lockdown


title: Src inhibitor reduces permeability without disturbing vascularization and prevents bone destruction in steroid-associated osteonecrotic lesions in rabbits

text: To examine the therapeutic effect of Src inhibitor on the VEGF mediating vascular hyperpermeability and bone destruction within steroid-associated osteonecrotic lesions in rabbits. Rabbits with high risk for progress to destructive repair in steroid-associated osteonecrosis were selected according to our published protocol. The selected rabbits were systemically administrated with either Anti-VEGF antibody (Anti-VEGF Group) or Src inhibitor (Src-Inhibition Group) or VEGF (VEGF-Supplement Group) or a combination of VEGF and Src inhibitor (Supplement & Inhibition Group) or control vehicle (Control Group) for 4 weeks. At 0, 2 and 4 weeks after administration, in vivo dynamic MRI, micro-CT based-angiography, histomorphometry and immunoblotting were employed to evaluate the vascular and skeletal events in different groups. The incidence of the destructive repair in the Anti-VEGF Group, Src-Inhibition Group and Supplement & Inhibition Group was all significantly lower than that in the Control Group. The angiogenesis was promoted in VEGF-Supplement Group, Src-Inhibition Group and Supplement & Inhibition Group, while the hyperpermeability was inhibited in Anti-VEGF Group, Src-Inhibition Group and Supplement & Inhibition Group. The trabecular structure was improved in Src-Inhibition Group and Supplement & Inhibition Group. Src inhibitor could reduce permeability without disturbing vascularization and prevent destructive repair in steroid-associated osteonecrosis.


“Urinary Tract Infection” in the Era of COVID‐19


title: Chapter 52 Gerbils

text: The chapter provides an overview of the taxonomy, history, and origin of the Mongolian gerbil. It provides quick, easy-to-use information on anatomy, physiology, and behavior as well as management and husbandry practices. It assists in the humane care and use of gerbils by including details on basic experimental methods for investigators, veterinary care, and the commonly seen diseases. Some more common uses of gerbils in research are described.


title: Respiratory and fecal shedding of Porcine respiratory coronavirus (PRCV) in sentinel weaned pigs and sequence of the partial S-gene of the PRCV isolates

text: Porcine respiratory coronavirus (PRCV), a spike (S) gene deletion mutant of Transmissible gastroenteritis virus (TGEV), causes mild or subclinical respiratory infections in pigs. The shedding of PRCV/TGEV was studied at different days post-arrival in fecal and nasal swabs from PRCV/TGEV seronegative sentinel pigs introduced into a PRCV seropositive herd with questionable TGEV serology and diarrhea. Nasal shedding of PRCV was detected in 57% and 63% of samples by nested-RT-PCR and cell culture immunofluorescence (CCIF), respectively. However fecal shedding of PRCV was detected in 37% of the samples by nested-RT-PCR and 19% by CCIF. Four respiratory and 5 fecal PRCV strains were isolated in swine testicle cells including nasal/fecal PRCV pairs (isolated at the same time) from 3 pigs. Comparison of nasal/fecal PRCV pairs from individual pigs revealed different deletions in the spike (S) gene (648 or 681 nt) in 2 pairs and a consistent change in nt 790/791 (aa T to V) for all pairs. In preliminary studies, inoculation of gnotobiotic pigs with each plaque-purified pair of the nasal and fecal PRCV isolates, revealed no clinical disease but different tropisms. The nasal isolate was shed both nasally and in feces, but the fecal isolate was shed only marginally in feces, and not nasally. Our results show that nested-RT-PCR was as sensitive as CCIF for PRCV detection in nasal swabs, but was more sensitive than CCIF for PRCV detection in fecal samples; alternatively PRCV shed in feces was more labile with loss of infectivity. The S-gene sequence differences found between the fecal and respiratory PRCV isolates may influence their tissue tropism. These new PRCV isolates should be useful to understand the molecular basis of coronavirus tropism and evolution in infected swine.