title: The Golgi associated ERI3 is a Flavivirus host factor

text: Dengue virus (DENV) is a mosquito-borne Flavivirus classified into four serotypes (DENV-1-4) that causes Dengue fever (DF), Dengue hemorrhagic Fever (DHF) or Dengue shock syndrome (DSS). An estimated 390 million people are at risk for infection with DENV and there are no effective vaccines or therapeutics. We utilized RNA chromatography coupled with quantitative mass spectrometry (qMS) to identify host RNA binding proteins (RBPs) that interact with DENV-2 RNA. We identified ERI3 (also PRNPIP and PINT1), a putative 3′–5′ RNA exonuclease, which preferentially associates with DENV-2 genomic RNA via interactions with dumbbell structures in the 3′ UTR. ERI3 is required for accumulation of DENV-2 genomic RNA and production of infectious particles. Furthermore, the mosquito homologue of ERI3 is required for DENV-2 replication in adult Aedes aegypti mosquitos implying that the requirement for ERI3 is conserved in both DENV hosts. In human cells ERI3 localizes to the Golgi in uninfected cells, but relocalizes near sites of DENV-2 replication in infected cells. ERI3 is not required for maintaining DENV-2 RNA stability or translation of the viral polyprotein, but is required for viral RNA synthesis. Our results define a specific role for ERI3 and highlight the importance of Golgi proteins in DENV-2 replication.


Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus, Asia


title: Differences in the origin of philosophy between Chinese medicine and western medicine: Exploration of the holistic advantages of Chinese medicine

text: To explore advantages of Chinese medicine (CM) by analyzing differences in the origin of philosophy for human health between CM and Western medicine (WM). Methodologically, a distinctive feature of CM is its systems theory, which is also the difference between CM and WM. Since the birth of CM, it has taken the human body as a whole from the key concepts of “qi, blood, yin-yang, viscera (Zang-Fu), and meridian and channel”, rather than a single cell or a particular organ. WM evolves from the Western philosophic way of thinking and merely uses natural sciences as the foundation. The development of WM is based on human structures, or anatomy, and therefore, research of WM is also based on the way of thinking of decomposing the whole human body into several independent parts, which is the impetus of promoting the development of WM. The core of CM includes the holistic view and the dialectical view. Chinese herbal medicines contain various components and treat a disease from multiple targets and links. Therefore, Chinese herbal medicines treat a diseased state by regulating and mobilizing the whole body rather than just regulating a single factor, since the diseased state is not only a problem in a local part of the body but a local reflection of imbalance of the whole body.


title: Etiology of respiratory disease in non-vaccinated, non-medicated calves in rearing herds

text: Abstract The aim of this study was to examine the occurrence of bacterial, mycoplasmal and viral pathogens in the lower respiratory tract of calves in all-in all-out calf-rearing units. According to clinical status, non-medicated calves with and without respiratory disease signs were selected of the 40 herds investigated to analyse the micro-organisms present in healthy and diseased calves. Tracheobronchial lavage (TBL) and paired serum samples were analysed for bacteria, mycoplasmas, respiratory syncytial virus (RSV), parainfluenza virus 3 (PIV3), bovine corona virus (BCV) and bovine adenovirus (BAV). Pasteurella multocida was the most common bacterial pathogen. It was isolated from 34% of the TBL samples in 28 herds and was associated with clinical respiratory disease (p <0.05) when other pathogenic bacteria or mycoplasma were present in the sample. Mannheimia spp. and Histophilus somni were rarely found. Mycoplasma bovis was not detected at all. Ureaplasma diversum was associated with clinical respiratory disease (p <0.05). TBL samples from healthy or suspect calves were more often negative in bacterial culture than samples from diseased calves (p <0.05). No viral infections were detected in six herds, while 16–21 herds had RSV, BCV, BAV or PIV3. In the herds that had calves seroconverted to BCV, respiratory shedding of BCV was more frequently observed than faecal shedding. This study showed that the microbial combinations behind BRD were diverse between herds. M. bovis, an emerging pathogen in many countries, was not detected.


title: Incidental Discovery of a COVID-19 Infection on a Reevaluation FDG PET/CT in a Patient Treated for Hodgkin Lymphoma

text: We report the results of (18)F-FDG PET/CT in an asymptomatic case of COVID-19 infection. A 27-year-old woman underwent FDG PET/CT for revaluation of a stage IIIE B Hodgkin lymphoma after the fourth cycle of chemotherapy. It showed intense avid FDG subpleural mixed ground-glass and consolidative lesions, especially in the left lung. Because of this morpho-metabolic aspect and the epidemic context, a viral pneumopathy was suspected. The patient who was initially asymptomatic was admitted for fever 28 hours after the PET/CT. The nasopharyngeal swab was positive for COVID-19, and the outcome was favorable.


title: The duration of cough in patients with H1N1 influenza

text: BACKGROUND: Cough is one of common symptoms of influenza, the cough duration and prevalence of postinfectious cough (PIC) after viral upper respiratory tract infection has not been well described. OBJECTIVES: We aim to investigate the duration of cough and prevalence of PIC and its relation with acute symptoms, airway inflammation and cough sensitivity in patients with H1N1 influenza. METHODS: Patients with acute symptoms of H1N1 influenza were enrolled and followed up until cough relived. Spirometry, induced sputum test, capsaicin challenge test were conducted in patients with PIC. Cough sensitivity was presented as logarithm of provocative concentration inducing five or more coughs (logC5). RESULTS: A total of 141 cases with H1N1 influenza were enrolled. In patients with H1N1 influenza, 97.2% of them complained cough. The duration of cough was as following: <1 week (73.0%); 1–2 weeks (7.8%); 2–3 weeks (7.8%); ≥3 weeks (8.5%). Twelve (8.5%) patients had cough lasting more than 3 weeks (PIC), 4 (2.8%) patients developed chronic cough (>8 weeks). Acute symptoms, spirometry, bronchial responsiveness and sputum differential cell count were similar between patients with PIC and those without PIC, however, there was a higher prevalence of previous PIC (58.3% vs 14.7%, P < 0.05) and elevated cough sensitivity (lgC5: 1.18 ± 0.58 vs 2.73 ± 0.33, P < 0.01) in patients with PIC as compared with the patients without PIC. CONCLUSIONS: Acute cough is common in patients with H1N1 PIC, only a few of patients develop chronic cough. Acute symptoms cannot predict PIC which is related with previous PIC and increased cough sensitivity.


title: Conceptualising the technical relationship of animal disease surveillance to intervention and mitigation as a basis for economic analysis

text: BACKGROUND: Surveillance and intervention are resource-using activities of strategies to mitigate the unwanted effects of disease. Resources are scarce, and allocating them to disease mitigation instead of other uses necessarily involves the loss of alternative sources of benefit to people. For society to obtain the maximum benefits from using resources, the gains from disease mitigation must be compared to the resource costs, guiding decisions made with the objective of achieving the optimal net outcome. DISCUSSION: Economics provides criteria to guide decisions aimed at optimising the net benefits from the use of scarce resources. Assessing the benefits of disease mitigation is no exception. However, the technical complexity of mitigation means that economic evaluation is not straightforward because of the technical relationship of surveillance to intervention. We argue that analysis of the magnitudes and distribution of benefits and costs for any given strategy, and hence the outcome in net terms, requires that mitigation is considered in three conceptually distinct stages. In Stage I, 'sustainment', the mitigation objective is to sustain a free or acceptable status by preventing an increase of a pathogen or eliminating it when it occurs. The role of surveillance is to document that the pathogen remains below a defined threshold, giving early warning of an increase in incidence or other significant changes in risk, and enabling early response. If a pathogen is not contained, the situation needs to be assessed as Stage II, 'investigation'. Here, surveillance obtains critical epidemiological information to decide on the appropriate intervention strategy to reduce or eradicate a disease in Stage III, 'implementation'. Stage III surveillance informs the choice, timing, and scale of interventions and documents the progress of interventions directed at prevalence reduction in the population. SUMMARY: This article originates from a research project to develop a conceptual framework and practical tool for the economic evaluation of surveillance. Exploring the technical relationship between mitigation as a source of economic value and surveillance and intervention as sources of economic cost is crucial. A framework linking the key technical relationships is proposed. Three conceptually distinct stages of mitigation are identified. Avian influenza, salmonella, and foot and mouth disease are presented to illustrate the framework.


title: Applications of omics approaches to the development of microbiological risk assessment using RNA virus dose–response models as a case study

text: T e in the amount of ‘omics’ data available and in our ability to interpret those data. The aim of this paper was to consider how omics techniques can be used to improve and refine microbiological risk assessment, using dose–response models for RNA viruses, with particular reference to norovirus through the oral route as the case study. The dose–response model for initial infection in the gastrointestinal tract is broken down into the component steps at the molecular level and the feasibility of assigning probabilities to each step assessed. The molecular mechanisms are not sufficiently well understood at present to enable quantitative estimation of probabilities on the basis of omics data. At present, the great strength of gene sequence data appears to be in giving information on the distribution and proportion of susceptible genotypes (for example due to the presence of the appropriate pathogen‐binding receptor) in the host population rather than in predicting specificities from the amino acid sequences concurrently obtained. The nature of the mutant spectrum in RNA viruses greatly complicates the application of omics approaches to the development of mechanistic dose–response models and prevents prediction of risks of disease progression (given infection has occurred) at the level of the individual host. However, molecular markers in the host and virus may enable more broad predictions to be made about the consequences of exposure in a population. In an alternative approach, comparing the results of deep sequencing of RNA viruses in the faeces/vomitus from donor humans with those from their infected recipients may enable direct estimates of the average probability of infection per virion to be made.


title: Myeloma care adaptations in the UK during SARS‐CoV‐2 pandemic: Challenges and measurable outcomes

text: Following the emergence of COVID-19 as a global pandemic, cancer clinicians have been faced with the challenge of continuing to manage patients, with limited data to inform practice (Poortmans et al 2020). Recently, a large prospective cohort study of 800 cancer patients diagnosed with symptomatic COVID-19 demonstrated a mortality rate of 28%, which was largely driven by age, co-morbidities and being a male (Lee et al 2020).


title: Chapter 10 Laboratory Diagnosis of Virus Diseases

text: Abstract An accurate virus diagnosis invariably requires laboratory testing of clinical specimens for the presence of virus, viral antigens, or specific antibodies. The past few decades have seen a major revolution in the operation of virus diagnostic laboratories and in their role in clinical patient management. Virus isolation has been largely replaced by sensitive nucleic acid detection assays and the measurement of specific antibodies at a very high level of sensitivity and specificity.


title: Virus Database and Online Inquiry System Based on Natural Vectors

text: We construct a virus database called VirusDB (http://yaulab.math.tsinghua.edu.cn/VirusDB/) and an online inquiry system to serve people who are interested in viral classification and prediction. The database stores all viral genomes, their corresponding natural vectors, and the classification information of the single/multiple-segmented viral reference sequences downloaded from National Center for Biotechnology Information. The online inquiry system serves the purpose of computing natural vectors and their distances based on submitted genomes, providing an online interface for accessing and using the database for viral classification and prediction, and back-end processes for automatic and manual updating of database content to synchronize with GenBank. Submitted genomes data in FASTA format will be carried out and the prediction results with 5 closest neighbors and their classifications will be returned by email. Considering the one-to-one correspondence between sequence and natural vector, time efficiency, and high accuracy, natural vector is a significant advance compared with alignment methods, which makes VirusDB a useful database in further research.


title: Recent Advances in Community-Acquired Pneumonia Inpatient and Outpatient

text: Community-acquired pneumonia (CAP) is a common illness, with the majority of patients treated out of the hospital, yet the greatest burden of the cost of care comes from inpatient management. In the past several years, the management of these patients has advanced, with new information about the natural history and prognosis of illness, the utility of serum markers to guide management, the use of appropriate clinical tools to guide the site-of-care decision, and the finding that guidelines can be developed in a way that improves patient outcome. The challenges to patient management include the emergence of new pathogens and the progression of antibiotic resistance in some of the common pathogens such as Streptococcus pneumoniae. Few new antimicrobial treatment options are available, and the utility of some new therapies has been limited by drug-related toxicity. Ancillary care for severe pneumonia with activated protein C and corticosteroids is being studied, but recently, inpatient care has been most affected by the development of evidence-based “core measures” for management that have been promoted by the Centers for Medicare and Medicaid Services, which form the basis for the public reporting of hospital performance in CAP care.


title: Human exhalation characterization with the aid of schlieren imaging technique

text: Abstract The purpose of this paper is to determine the dispersion and distribution characteristics of exhaled airflow for accurate prediction of disease transmission. The development of airflow dynamics of human exhalation was characterized using nonhazardous schlieren photography technique, providing a visualization and quantification of turbulent exhaled airflow from 18 healthy human subjects whilst standing and lying. The flow shape of each breathing pattern was characterized by two angles and averaged values of 18 subjects. Two exhaled air velocities, u m and u p , were measured and compared. The mean peak centerline velocity, u m was found to decay correspondingly with increasing horizontal distance x in a form of power function. The mean propagation velocity, u p was found to correlate with physiological parameters of human subjects. This was always lower than u m at the mouth/nose opening, due to a vortex like airflow in front of a single exhalation cycle. When examining the talking and breathing process between two persons, the potential infectious risk was found to depend on their breathing patterns and spatial distribution of their exhaled air. Our study when combined with information on generation and distributions of pathogens could provide a prediction method and control strategy to minimize infection risk between persons in indoor environments.


title: Enhanced Expression of Rabies Virus Surface G-Protein in Escherichia coli using SUMO Fusion

text: Fusion systems are known to increase the expression of difficult to express recombinant proteins in soluble form to facilitate their purification. Rabies glycoprotein was also tough to express at sufficient level in soluble form in both E. coli and plant. The present work was aimed to over-express and purify this membrane protein from soluble extract of E. coli. Fusion of Small Ubiqutin like Modifier (SUMO) with rabies glycoprotein increased ~1.5 fold higher expression and ~3.0 fold solubility in comparison to non-fused in E. coli. The SUMO fusion also simplified the purification process. Previously engineered rabies glycoprotein gene in tobacco plants provides complete protection to mice, but the expression was very low for purification. Our finding demonstrated that the SUMO-fusion was useful for enhancing expression and solubility of the membrane protein and again proves to be a good alternative technology for applications in biomedical and pharmaceutical research. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10930-011-9373-6) contains supplementary material, which is available to authorized users.


title: Cellular DDX3 regulates Japanese encephalitis virus replication by interacting with viral un-translated regions

text: Abstract Japanese encephalitis virus is one of the most common causes for epidemic viral encephalitis in humans and animals. Herein we demonstrated that cellular helicase DDX3 is involved in JEV replication. DDX3 knockdown inhibits JEV replication. The helicase activity of DDX3 is crucial for JEV replication. GST-pulldown and co-immunoprecipitation experiments demonstrated that DDX3 could interact with JEV non-structural proteins 3 and 5. Co-immunoprecipitation and confocal microscopy analysis confirmed that DDX3 interacts and colocalizes with these viral proteins and viral RNA during the infection. We determined that DDX3 binds to JEV 5′ and 3′ un-translated regions. We used a JEV-replicon system to demonstrate that DDX3 positively regulates viral RNA translation, which might affect viral RNA replication at the late stage of virus infection. Collectively, we identified that DDX3 is necessary for JEV infection, suggesting that DDX3 might be a novel target to design new antiviral agents against JEV or other flavivirus infections.


title: Estimating the prevalence and risk of COVID-19 among international travelers and evacuees of Wuhan through modeling and case reports

text: Coronavirus disease 2019 (COVID-19) started in Wuhan, China and has spread through other provinces and countries through infected travelers. On January 23(rd), 2020, China issued a quarantine and travel ban on Wuhan because travelers from Wuhan were thought to account for the majority of exported COVID-19 cases to other countries. Additionally, countries evacuated their citizens from Wuhan after institution of the travel ban. Together, these two populations account for the vast majority of the “total cases with travel history to China” as designated by the World Health Organization (WHO). The current study aims to assess the prevalence and risk of COVID-19 among international travelers and evacuees of Wuhan. We first used case reports from Japan, Singapore, and Korea to investigate the date of flights of infected travelers. We then used airline traveler data and the number of infected exported cases to correlate the cases with the number of travelers for multiple countries. Our findings suggest that the risk of COVID-19 infection is highest among Wuhan travelers between January 19(th) and 22(nd), 2020, with an approximate infection rate of up to 1.3% among international travelers. We also observed that evacuee infection rates varied heavily between countries and propose that the timing of the evacuation and COVID-19 testing of asymptomatic evacuees played significant roles in the infection rates among evacuees. These findings suggest COVID-19 cases and infectivity are much higher than previous estimates, including numbers from the WHO and the literature, and that some estimates of the infectivity of COVID-19 may need re-assessment.


title: Skin involvement in SARS‐CoV‐2 infection: Case series

text: since the beginning of the well-known SARS CoV-2 pandemic, skin involvement both in adults and children has been signaled (1,2). However, large-scale studies describing skin manifestations and their etiopathogenic correlation with COVID-19 in detail have not been reported yet. Through the analysis of emerging data from literature (3) and the direct observation of three patients with COVID-19 (SARS-CoV-2 detection from nasopharyngeal swab samples through RNA extraction) and dermatological manifestations, we have hypothesized different mechanisms for their development. This article is protected by copyright. All rights reserved.


title: Identification of the relative timing of infectiousness and symptom onset for outbreak control

text: In an outbreak of an emerging disease the epidemiological characteristics of the pathogen may be largely unknown. A key determinant of ability to control the outbreak is the relative timing of infectiousness and symptom onset. We provide a method for identifying this relationship with high accuracy based on data from household-stratified symptom-onset data. Further, this can be achieved with observations taken on only a few specific days, chosen optimally, within each household. This constitutes an important tool for outbreak response. An accurate and computationally-efficient heuristic for determining the optimal surveillance scheme is introduced. This heuristic provides a novel approach to optimal design for Bayesian model discrimination.


title: Genetics, Recombination and Clinical Features of Human Rhinovirus Species C (HRV-C) Infections; Interactions of HRV-C with Other Respiratory Viruses

text: To estimate the frequency, molecular epidemiological and clinical associations of infection with the newly described species C variants of human rhinoviruses (HRV), 3243 diagnostic respiratory samples referred for diagnostic testing in Edinburgh were screened using a VP4-encoding region-based selective polymerase chain reaction (PCR) for HRV-C along with parallel PCR testing for 13 other respiratory viruses. HRV-C was the third most frequently detected behind respiratory syncytial virus (RSV) and adenovirus, with 141 infection episodes detected among 1885 subjects over 13 months (7.5%). Infections predominantly targeted the very young (median age 6–12 months; 80% of infections in those <2 years), occurred throughout the year but with peak incidence in early winter months. HRV-C was detected significantly more frequently among subjects with lower (LRT) and upper respiratory tract (URT) disease than controls without respiratory symptoms; HRV-C mono-infections were the second most frequently detected virus (behind RSV) in both disease presentations (6.9% and 7.8% of all cases respectively). HRV variants were classified by VP4/VP2 sequencing into 39 genotypically defined types, increasing the current total worldwide to 60. Through sequence comparisons of the 5′untranslated region (5′UTR), the majority grouped with species A (n = 96; 68%, described as HRV-Ca), the remainder forming a phylogenetically distinct 5′UTR group (HRV-Cc). Multiple and bidirectional recombination events between HRV-Ca and HRV-Cc variants and with HRV species A represents the most parsimonious explanation for their interspersed phylogeny relationships in the VP4/VP2-encoding region. No difference in age distribution, seasonality or disease associations was identified between HRV-Ca and HRV-Cc variants. HRV-C-infected subjects showed markedly reduced detection frequencies of RSV and other respiratory viruses, providing evidence for a major interfering effect of HRV-C on susceptibility to other respiratory virus infections. HRV-C's disease associations, its prevalence and evidence for interfering effects on other respiratory viruses mandates incorporation of rhinoviruses into future diagnostic virology screening.


title: Application of chicken egg yolk immunoglobulins in the control of terrestrial and aquatic animal diseases: A review

text: Abstract Oral administration of chicken egg yolk immunoglobulin (IgY) has attracted considerable attention as a means of controlling infectious diseases of bacterial and viral origin. Oral administration of IgY possesses many advantages compared with mammalian IgG including cost-effectiveness, convenience and high yield. This review presents an overview of the potential to use IgY immunotherapy for the prevention and treatment of terrestrial and aquatic animal diseases and speculates on the future of IgY technology. Included are a review of the potential application of IgY for the treatment of livestock diseases such as mastitis and diarrhea, poultry diseases such as Salmonella, Campylobacteriosis, infectious bursal disease and Newcastle disease, as well as aquatic diseases like shrimp white spot syndrome virus, Yersina ruckeri and Edwardsiella tarda. Some potential obstacles to the adoption of IgY technology are also discussed.


title: VALORACIÓN MEDICOLEGAL DE LA INFECCIÓN POR COVID-19 EN EL ÁMBITO LABORAL

text: Resumen España ha sido uno de los países con mayor afectación por la pandemia COVID-19. Hasta la fecha, un 22% de los casos notificados son personal sanitario, siendo significativamente mayor este porcentaje entre las mujeres (76% del personal sanitario contagiado). Ante la evidencia de que muchos profesionales sanitarios pueden haberse contagiado en su puesto de trabajo, hemos revisado la problemática en el ámbito de la salud laboral que produce el COVID- 19. Se describen los decretos publicados por el Gobierno de España y las recomendaciones de la Organización Internacional del Trabajo. Concretamente se analiza la clasificación del SARS-CoV-2 como agente biológico del grupo 3 y su calificación como accidente de trabajo o enfermedad profesional, y sus repercusiones en las situaciones de incapacidades (temporales y permanentes), secuelas y muerte; así como el estudio de las posibles demandas de aquellos que lo han sufrido como consecuencia de falta de medidas preventivas. Abstract Spain is one of the countries most affected by the COVID-19 pandemic. To date, 22% of reported cases are healthcare professionals, and 76% of them are women. Given that many healthcare professionals may have been infected in their workplace, we have reviewed the occupational health challenges posed by COVID-19. The Spanish laws and the recommendations of the International Labor Organization are described. The review covers the classification of SARS-CoV-2 as a biological agent in group 3 and of COVID-19 as an occupational accident or disease, its repercussions in temporary and permanent disabilities, sequelae and death, and the possible claims of those who may have developed the disease due to the lack of preventive measures.


COVID-19 experience reports: An emerging trend in scholarly communication


title: Off‐label Use of Tocilizumab in Patients with SARS‐CoV‐2 Infection

text: The spread of the novel‐Coronavirus infection worldwide represents a challenge for physicians. Particularly, no approved therapy has demonstrated to have an impact in treating patients who develop severe respiratory insufficiency so far. The use of humanized anti‐human inteleukine‐6 receptor antibody tocilizumab seems a promising strategy for these patients. This article is protected by copyright. All rights reserved.


title: Language Barriers and the Management of Bronchiolitis in a Pediatric Emergency Department

text: OBJECTIVE: Language barriers may influence the management of pediatric emergency department (PED) patients who may not align with evidence-based guidelines from the American Academy of Pediatrics. Our objective was to determine if a family's preferred language of Spanish versus English was associated with differences in management of bronchiolitis in the PED. METHODS: We conducted a retrospective study of children ≤2 years old diagnosed with bronchiolitis in a PED over a 7-year period. Rates of PED testing, interventions, and disposition among children whose families’ preferred language was Spanish were compared to children whose families’ preferred language was English. Primary outcomes were frequencies of chest x-ray and bronchodilator orders. Secondary outcomes were diagnostic testing, medication orders, and disposition. Logistic regression was used to calculate adjusted odds ratios after controlling for age, emergency severity index, prior visit, and nesting within attending physicians. RESULTS: A total of 13,612 encounters were included. Spanish-speaking families were more likely to have chest x-rays (35.8% vs 26.7%, P < .0001; adjusted odds ratio [aOR] 1.5; 95% confidence interval [CI] 1.2–1.9), complete blood counts (8.2% vs 4.9%, P < .005; aOR 1.7; 95% CI 1.2–2.5), and blood cultures ordered (8.1% vs 5.0%, P < .05; aOR 1.7; 95% CI 1.2–2.4). No other differences in bronchodilators, medication orders, or disposition were found between the 2 groups. CONCLUSIONS: Among children diagnosed with bronchiolitis, Spanish-speaking families were more likely to have chest x-rays, complete blood counts, and blood cultures ordered compared to English-speaking families. Further research on how clinical practice guidelines and equity-focused guidelines can impact disparities in diagnostic testing within the PED is warranted.


title: Changing paradigms in sinus and skull base surgery as the COVID‐19 pandemic evolves: Preliminary experience from a single Italian tertiary care center

text: BACKGROUND: Italy was the first European country suffering from COVID‐19. Health care resources were redirected to manage the pandemic. We present our initial experience with the management of urgent and nondeferrable surgeries for sinus and skull base diseases during the COVID‐19 pandemic. METHODS: A retrospective review of patients treated in a single referral center during the first 2 months of the pandemic was performed. A comparison between the last 2‐month period and the same period of the previous year was carried out. RESULTS: Twenty‐four patients fulfilled the inclusion criteria. A reduction of surgical activity was observed (−60.7%). A statistically significant difference in pathologies treated was found (P = .016), with malignancies being the most frequent indication for surgery (45.8%). CONCLUSIONS: Although we feel optimistic for the future, we do not feel it is already time to restart elective surgeries. Our experience may serve for other centers who are facing the same challenges.


title: Dendritic Cell-Specific Delivery of Flt3L by Coronavirus Vectors Secures Induction of Therapeutic Antitumor Immunity

text: Efficacy of antitumor vaccination depends to a large extent on antigen targeting to dendritic cells (DCs). Here, we assessed antitumor immunity induced by attenuated coronavirus vectors which exclusively target DCs in vivo and express either lymphocyte- or DC-activating cytokines in combination with a GFP-tagged model antigen. Tracking of in vivo transduced DCs revealed that vectors encoding for Fms-like tyrosine kinase 3 ligand (Flt3L) exhibited a higher capacity to induce DC maturation compared to vectors delivering IL-2 or IL-15. Moreover, Flt3L vectors more efficiently induced tumor-specific CD8(+) T cells, expanded the epitope repertoire, and provided both prophylactic and therapeutic tumor immunity. In contrast, IL-2- or IL-15-encoding vectors showed a substantially lower efficacy in CD8(+) T cell priming and failed to protect the host once tumors had been established. Thus, specific in vivo targeting of DCs with coronavirus vectors in conjunction with appropriate conditioning of the microenvironment through Flt3L represents an efficient strategy for the generation of therapeutic antitumor immunity.


title: An Overview of Typical Infections of Research Mice: Health Monitoring and Prevention of Infection

text: There are many reasons to keep research mice healthy and free from infections. The two most important of these are to protect the health and welfare of research mice and to prevent infections from negatively impacting research. Just as the genetic integrity of a mouse strain will influence the reproducibility and validity of research data, so too will the microbiologic integrity of the animals. This has been repeatedly demonstrated in the literature of laboratory animal sciences wherein the direct impact of infections on physiologic parameters under study have been described. Therefore, it is of great importance that scientists pay close attention to the health status of their research animal colonies and maintain good communications with the animal facility personnel at their institution about mouse health issues. This overview provides information about animal health monitoring (HM) in research mouse colonies including commonly monitored agents, diagnostic methods, HM program, risk assessment, and animal facility biosecurity. Lastly, matters of communication with laboratory animal professionals at research institutions are also addressed. © 2015 by John Wiley & Sons, Inc.


title: Determining the Cause of Death Among Children Hospitalized With Respiratory Illness in Kenya: Protocol for Pediatric Respiratory Etiology Surveillance Study (PRESS)

text: BACKGROUND: In sub-Saharan Africa, where the burden of respiratory disease–related deaths is the highest, information on the cause of death remains inadequate because of poor access to health care and limited availability of diagnostic tools. Postmortem examination can aid in the ascertainment of causes of death. This manuscript describes the study protocol for the Pediatric Respiratory Etiology Surveillance Study (PRESS). OBJECTIVE: This study protocol aims to identify causes and etiologies associated with respiratory disease–related deaths among children (age 1-59 months) with respiratory illness admitted to the Kenyatta National Hospital (KNH), the largest public hospital in Kenya, through postmortem examination coupled with innovative approaches to laboratory investigation. METHODS: We prospectively followed children hospitalized with respiratory illness until the end of clinical care or death. In case of death, parents or guardians were offered grief counseling, and postmortem examination was offered. Lung tissue specimens were collected using minimally invasive tissue sampling and conventional autopsy where other tissues were collected. Tissues were tested using histopathology, immunohistochemistry, and multipathogen molecular-based assays to identify pathogens. For each case, clinical and laboratory data were reviewed by a team of pathologists, clinicians, laboratorians, and epidemiologists to assign a cause of and etiology associated with death. RESULTS: We have enrolled pediatric cases of respiratory illness hospitalized at the KNH at the time of this submission; of those, 14.8% (140/945) died while in the hospital. Both analysis and interpretation of laboratory results and writing up of findings are expected in 2019-2020. CONCLUSIONS: Postmortem studies can help identify major pathogens contributing to respiratory-associated deaths in children. This information is needed to develop evidence-based prevention and treatment policies that target important causes of pediatric respiratory mortality and assist with the prioritization of local resources. Furthermore, PRESS can provide insights into the interpretation of results using multipathogen testing platforms in resource-limited settings. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/10854


title: Computational studies for the structure and function of mRPE65

text: Abstract The mRPE65 protein is one form of the RPE65 protein and plays a very important role in the visual cycle. However, its 3D structure and detailed mechanism of function are still unclear because of difficulties with isolation and crystallization. This computational study reports a model for the mRPE65 protein structure derived from a model for sRPE65. The natural substrate for RPE65 has been shown to be a retinyl ester and, by utilizing the Autodock and the Ligplot programs, the interactions between the ester and the protein as well as the effects of several mutations on these interactions are studied. Finally, the position of the binding site is proposed based on an iterative process and the effects of the mutations on the binding site are also discussed.


title: Inequalities in access to water and soap matter for the COVID-19 response in sub-Saharan Africa

text: The COVID-19 pandemic has spread rapidly since the first case notification of the WHO in December 2019. Lacking an effective treatment, countries have implemented non-pharmaceutical interventions including social distancing measures and have encouraged maintaining adequate and frequent hand hygiene to slow down the disease transmission. Although access to clean water and soap is universal in high-income settings, it remains a basic need many do not have in low- and middle-income settings. We analyzed data from Demographic and Health Surveys (DHS) of 16 countries in sub-Saharan Africa, using the most recent survey since 2015. Differences in the percentage of households with an observed handwashing place with water and soap were estimated by place of residence and wealth quintiles. Equiplots showed wide within-country disparities, disproportionately affecting the poorest households and rural residents, who represent the majority of the population in most of the countries. Social inequalities in access to water and soap matter for the COVID-19 response in sub-Saharan Africa. Interventions such as mass distribution of soap and ensuring access to clean water, along with other preventive strategies should be scaled up to reach the most vulnerable populations.


title: Isolation and characterization of chicken lung mesenchymal stromal cells and their susceptibility to avian influenza virus

text: Abstract In this study, we isolated and characterized mesenchymal stromal cells (MSCs) from the lungs of 1- to 2-week-old chickens. Microscopically, the cultured cells showed fibroblast-like morphology. Phenotypically these cells expressed CD44, CD90, CD105 and the transcription factor PouV, which has been shown to be critical for stem cell self-renewal and pluripotency. The multipotency of chicken MSCs was demonstrated by their ability to undergo adipogenic and osteogenic differentiation. Like chicken bone marrow MSCs and mammalian MSCs, chicken lung MSCs had immunoregulatory activity and profoundly suppressed the proliferative capacity of T cells in response to a mitogenic stimulus. Next, we examined the susceptibility of these cells to H1N1 and H9N5 avian influenza (AI) viruses. The lung MSCs were shown to express known influenza virus alpha-2,3 and alpha-2,6 sialic acid receptors and to support replication of both the avian H1N1 and avian H9N5 influenza strains. Viral infection of MSCs resulted in cell lysis and cytokine and chemokine production. Further characterization of lung MSCs in chicken and other mammalian species may help in understanding the pathogenesis of infectious and non-infectious lung diseases and the mechanisms of lung injury repair.


title: α 1-Antitrypsin and the serpins: variation and countervariation

text: Abstract α 1-Antitrypsin is a plasma protein which protects elastic tissue from proteolytic attack. Consequently, genetic deficiency, or the oxidation of its reactive centre in cigarette smokers can result in the degenerative lung disease emphysema. Structural studies explain the mechanisms involved and have also drawn attention to a new family of serine proteinase inhibitors. The specificity of each of these inhibitors is primarily dependent on a single amino acid at its reactive centre. Site-directed mutagenesis is enabling the production of specifically designed inhibitors for therapeutic use, including an improved replacement for α 1-antitrypsin deficiency.


title: Convalescent Plasma, an Apheresis Research Project Targeting and Motivating the Fully Recovered COVID 19 Patients: A Rousing Message of Clinical Benefit To Both Donors and Recipients Alike

text: This concise manuscript aims to make suggestions for a small step forward in both preventative and therapeutic measures against the Coronavirus disease 2019 (COVID 19) pandemic. This targeted strategy consists of using fully recovered COVID 19 Heroes, that is, brave volunteers, as the source of antibodies in plasma collected by plasmapheresis or affinity column- derived antibodies, both are sterilised and pathogen inactivated for substitution therapy for use in those populations in need of antibody. This include for use in critically ill COVID 19 patients and as a preventative measure, in those at potential risk of infection as no vaccine is yet available. This would be a small step forward, while we are waiting to produce an effective, validated vaccine and witnessing increasing demands for testing and self-isolation which are the two most effective current strategies. In line with this concept, some methodological aspects of the use of the UVC sterilization of FFP/ cryoprecipitate-depleted FFP or immunoglobulins containing neutralizing antibodies for clinical use against COVID-19 are highlighted. The plasmapheresis procedure is, of course, particularly targeted to male donors, who consist of about 75 % of the COVID-19 population and who are able to undergo multiple double, or even triple plasmapheresis procedures. Moreover, as some of these donors have already been in an induced-hypercoagulable state and prone to thrombosis and DVT, this strategy will be partially aimed at improving their health with the use of citrate based anticoagulants and removal of high molecular weight viscous components which contribute to the untoward clinical effects of DVT. Repeated targeted plasmapheresis or plasma exchange of selected COVID-19 positive individuals would undoubtedly lower their state of hypercoagulability and normalize their hypercoagulability. The recipients of such a derived FFP-product would benefit from the two to 3 doses of viral inactivated antibodies, which could neutralize the viral antigens even at very low concentration if present in the early stage. So, this practice would be a double-edged sword with benefits for both donors and recipients.


New Perspectives from Misdiagnosis: A Case of Primary Cutaneous Cryptococcosis Treated with Hydroxychloroquine Sulfate Successfully


Children are at risk from COVID-19


title: Ventilation Techniques and Risk for Transmission of Coronavirus Disease, Including COVID-19: A Living Systematic Review of Multiple Streams of Evidence

text: BACKGROUND: Mechanical ventilation is used to treat respiratory failure in coronavirus disease 2019 (COVID-19). PURPOSE: To review multiple streams of evidence regarding the benefits and harms of ventilation techniques for coronavirus infections, including that causing COVID-19. (PROSPERO registration: CRD42020178187) DATA SOURCES: 21 standard, World Health Organization–specific and COVID-19–specific databases, without language restrictions, until 1 May 2020. STUDY SELECTION: Studies of any design and language comparing different oxygenation approaches in patients with coronavirus infections, including severe acute respiratory syndrome (SARS) or Middle East respiratory syndrome (MERS), or with hypoxemic respiratory failure. Animal, mechanistic, laboratory, and preclinical evidence was gathered regarding aerosol dispersion of coronavirus. Studies evaluating risk for virus transmission to health care workers from aerosol-generating procedures (AGPs) were included. DATA EXTRACTION: Independent and duplicate screening, data abstraction, and risk of bias assessment (GRADE for certainty of evidence and AMSTAR 2 for included systematic reviews). DATA SYNTHESIS: 123 studies were eligible (45 on COVID-19, 70 on SARS, 8 on MERS), but only 5 studies (1 on COVID-19, 3 on SARS, 1 on MERS) adjusted for important confounders. A study in hospitalized patients with COVID-19 reported slightly higher mortality with noninvasive ventilation (NIV) than with invasive mechanical ventilation (IMV), but 2 opposing studies, 1 in patients with MERS and 1 in patients with SARS, suggest a reduction in mortality with NIV (very low-certainty evidence). Two studies in patients with SARS report a reduction in mortality with NIV compared with no mechanical ventilation (low-certainty evidence). Two systematic reviews suggest a large reduction in mortality with NIV compared with conventional oxygen therapy. Other included studies suggest increased odds of transmission from AGPs. LIMITATION: Direct studies in COVID-19 are limited and poorly reported. CONCLUSION: Indirect and low-certainty evidence suggests that use of NIV, similar to IMV, probably reduces mortality but may increase the risk for transmission of COVID-19 to health care workers. PRIMARY FUNDING SOURCE: World Health Organization.


title: Renal Complications and Their Prognosis in Korean Patients with Middle East Respiratory Syndrome-Coronavirus from the Central MERS-CoV Designated Hospital

text: Some cases of Middle East Respiratory Syndrome-Coronavirus (MERS-CoV) infection presented renal function impairment after the first MERS-CoV patient died of progressive respiratory and renal failure. Thus, MERS-CoV may include kidney tropism. However, reports about the natural courses of MERS-CoV infection in terms of renal complications are scarce. We examined 30 MERS-CoV patients admitted to National Medical Center, Korea. We conducted a retrospective analysis of the serum creatinine (SCr), estimated glomerular filtration rate (eGFR), urine dipstick tests, urinary protein quantitation (ACR or PCR), and other clinical parameters in all patients. Two consecutive results of more than trace (or 1+) of albumin and blood on dipstick test occurred in 18 (60%) (12 [40%]) and 22 (73.3%) (19 [63.3%]) patients, respectively. Fifteen (50.0%) patients showed a random urine ACR or PCR more than 100 mg/g Cr. Eight (26.7%) patients showed acute kidney injury (AKI), and the mean and median durations to the occurrence of AKI from symptom onset were 18 and 16 days, respectively. Old age was associated with a higher occurrence of AKI in the univariate analysis (HR [95% CI]: 1.069 [1.013-1.128], P = 0.016) and remained a significant predictor of the occurrence of AKI after adjustment for comorbidities and the application of a mechanical ventilator. Diabetes, AKI, and the application of a continuous renal replacement therapy (CRRT) were risk factors for mortality in the univariate analysis (HR [95% CI]: diabetes; 10.133 [1.692-60.697], AKI; 12.744 [1.418-114.565], CRRT; 10.254 [1.626-64.666], respectively). Here, we report renal complications and their prognosis in 30 Korean patients with MERS-CoV.


title: COVID‐19 and Indigenous Peoples: An imperative for action

text: Every person on Earth has been affected in some way by the coronavirus disease (COVID-19) pandemic. However, there is a marked inequity in the impact and threat of the disease for the 370 million Indigenous Peoples worldwide. While honouring diversity in peoples and cultures, this editorial (written by a collaborative of Indigenous nurses from Australia, Aotearoa (New Zealand), Canada, the United States of America and Central America), explores contemporary issues raised for Indigenous communities by this latest public health emergency. Please note, while we may describe a situation about a specific Indigenous group, readers can be assured that the issues we raise are endemic across colonised Indigenous communities globally.


title: Oxidative stress influences positive strand RNA virus genome synthesis and capping

text: Abstract Flaviviruses are 5′ capped positive-stranded RNA viruses that replicate their genomes within endoplasmic reticulum-derived vesicles. Flaviviruses are well known to induce oxidative stress late in infection but it is unknown if oxidative stress plays a positive role in the viral RNA replication cycle. We therefore examined how oxidation affects flavivirus RNA replication. We found that antioxidant treatment reduced virus production, reduced the viral positive-to-negative strand RNA ratio, and resulted in the accumulation of uncapped positive-sense viral RNAs. Treatment of the NS5 RNA capping enzyme in vitro with oxidizing agents enhanced guanylyltransferase activity, indicating that the guanylyltransferase function of the flavivirus NS5 RNA capping enzyme is activated by oxidative conditions. Antioxidant treatment also reduced alphavirus RNA replication and protein expression while enhancing nsP1 capping activity. These findings suggest that RNA viruses may utilize oxidative stress induced during infection to help temporally control genome RNA capping and genome replication.


title: Separation methods used for Scutellaria baicalensis active components

text: Abstract Scutellaria baicalensis Georgi is one of the most widely used traditional Chinese herbal medicines. Its roots have been used for anti-inflammation, anticancer, antiviral and antibacterial infections of the respiratory and the gastrointestinal tract, cleaning away heat, moistening aridity, purging fire, detoxifying toxicosis, reducing the total cholesterol level and decreasing blood pressures. Baicalin, baicalein, wogonin and oroxylin A are its main active components. This review provides an overview of various separation, detection, and identification techniques employed for the quantitative and qualitative determination of these active components. Applications of high-performance liquid chromatography, high-speed counter-current chromatography, thin layer chromatography, capillary electrophoresis, and micellar electrokinetic capillary chromatography to the separation and determination of these active components are described. Examples of identification of these active components and their metabolites in complex matrices by high-performance liquid chromatography–mass spectrometry and gas chromatography–mass spectrometry are also presented. The advantages and limitations of these separation and identification methods are assessed and discussed.


title: Silver Lining of COVID-19: heightened global interest in pneumococcal and influenza vaccines, an infodemiology study

text: BACKGROUND: Health-seeking behaviors change during pandemics and may increase with regard to illnesses with symptoms similar to the pandemic. The global reaction to COVID-19 may drive interest in vaccines for other diseases. OBJECTIVES: Our study investigated the correlation between global online interest in COVID-19 and interest in CDC-recommended routine vaccines. DESIGN, SETTINGS, MEASUREMENTS: This infodemiology study used Google Trends data to quantify worldwide interest in COVID-19 and CDC-recommended vaccines using the unit search volume index (SVI), which estimates volume of online search activity relative to highest volume of searches within a specified period. SVIs from December 30, 2019 to March 30, 2020 were collected for “coronavirus (Virus)” and compared with SVIs of search terms related to CDC-recommended adult vaccines. To account for seasonal variation, we compared SVIs from December 30, 2019 to March 30, 2020 with SVIs from the same months in 2015 to 2019. We performed country-level analyses in ten COVID-19 hotspots and ten countries with low disease burden. RESULTS: There were significant positive correlations between SVIs for “coronavirus (Virus)” and search terms for pneumococcal (R=0.89, p<0.0001) and influenza vaccines (R=0.93, p<0.0001) in 2020, which were greater than SVIs for the same terms in 2015-2019 (p= 0.005, p<0.0001, respectively). Eight in ten COVID-19 hotspots demonstrated significant positive correlations between SVIs for coronavirus and search terms for pneumococcal and influenza vaccines. LIMITATIONS: SVIs estimate relative changes in online interest and do not represent for people with no Internet access. CONCLUSION: A peak in worldwide interest in pneumococcal and influenza vaccines coincided with the COVID-19 pandemic in February and March 2020. Trends are likely not seasonal in origin and may be driven by Covid-19 hotspots. Global events may change public perception about the importance of vaccines. Our findings may herald higher demand for pneumonia and influenza vaccines in the upcoming season.


title: COVID-19: Soziale Arbeit auf der Suche nach ihrem Auftrag im Katastrophenfall

text: COVID-19 stellt die Soziale Arbeit vor neue und alte Herausforderungen. Die Pandemie macht den Ausschluss marginalisierter Personengruppen und globale Ungleichheiten besonders sichtbar. Der Beitrag versteht die Corona-Pandemie als „Katastrophe“ und stellt erste Überlegungen dazu an, wie Soziale Arbeit ihren gesellschaftlichen Auftrag angesichts von COVID-19 ausgestalten und sich nachhaltig in der Katastrophenhilfe verankern kann.


title: Chapter 2 Classification of Viruses and Phylogenetic Relationships

text: Abstract The taxonomy of viruses represents a unique classification system that recognizes boundaries among at first sight a continuum of properties. Genome sequencing has brought into sharp debate the origin of viruses, with RNA viruses perhaps having a separate evolutionary lineage. The criteria adopted for assessing the causal linkage between virus and disease deviates from those normally adopted for bacterial diseases and reflects those unique properties that underlie the principles of virus classification.


title: Controlling Nipah virus encephalitis in Bangladesh: Policy options

text: Nipah virus (NiV) encephalitis is endemic in Bangladesh, with yearly seasonal outbreaks occurring since 2003. NiV has a notable case fatality rate, 75–100 per cent depending on the strain. In Bangladesh, primary transmission to humans is believed to be because of consumption of bat-contaminated date palm sap (DPS). Both the disease and the virus have been investigated extensively, however efforts to implement preventive strategies have met social and cultural challenges. Here we present a variety of community approaches to control the spread of Nipah encephalitis, along with advantages and disadvantages of each. This information may be useful to health workers and policymakers in potential NiV outbreak areas in Southeast Asia.


Chloroquine and hydroxychloroquine during pregnancy: what do we know?


Citoyens de la planète contre le redoutable SARS-CoV-2


title: A New Measure for Assessing the Public Health Response to a Middle East Respiratory Syndrome Coronavirus Outbreak

text: Contact monitoring is an essential component of the public health response to a Middle East respiratory syndrome coronavirus outbreak, and is required for an effective quarantine to contain the epidemic. The timeliness of a quarantine is associated with its effectiveness. This paper provides a conceptual framework to describe the process of contact monitoring, and proposes a new measure called the “timely quarantined proportion” as a tool to assess the adequacy of a public health response.


title: COVID19: a case report of thrombus in transit

text: BACKGROUND: The global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused significant morbidity and mortality, not only through devastating lung injury, but also due to multiple malfunctions in the cardiovascular system. The primary aetiology is believed to be mediated through lung alveolar injury; however, a few published reports have linked SARS-CoV-2 to significant organ dysfunction, venous thrombo-embolism, and coagulopathy. In view of the fact that the utility of tissue plasminogen activator in this population is not well studied, we present this case of rapid improvement in oxygenation after successful lytic therapy for thrombus in transit in this patient with SARS-CoV-2. CASE SUMMARY: We discuss a patient admitted with SARS-CoV-2 pneumonia. Due to the development of dramatic hypoxia, he underwent echocardiography which demonstrated extensive thrombus in transit. He received successful thrombolytic therapy with tissue plasminogen activator, with subsequent improvement in oxygenation. The patient was successfully discharged home on 2 L of oxygen via nasal cannula, and continues to improve at follow-up with his cardiologist and primary care physician. CONCLUSION: This case not only highlights embolic causes of hypoxia in SARS-CoV-2, but demonstrates the important utility of an echocardiogram and tissue plasminogen activator in this population.


title: Respiratory infections and asthma

text: What we know What we need to know


title: Effects of a mixed media education intervention program on increasing knowledge, attitude, and compliance with standard precautions among nursing students: A randomized controlled trial

text: BACKGROUND: Standard precautions (SPs) are considered fundamental protective measures to manage health care-associated infections and to reduce occupational health hazards. This study intended to assess the effectiveness of a mixed media education intervention to enhance nursing students' knowledge, attitude, and compliance with SPs. METHOD: A randomized controlled trial with 84 nursing students was conducted in a teaching hospital in Hubei, China. The intervention group (n = 42) attended 3 biweekly mixed media education sessions, consisting of lectures, videos, role-play, and feedback with 15-20 minutes of individual online supervision and feedback sessions following each class. The control group learned the same material through self-directed readings. Pre- and posttest assessments of knowledge, attitudes, and compliance were assessed with the Knowledge with Standard Precautions Questionnaire, Attitude with Standard Precautions Scale, and the Compliance with Standard Precautions Scale, respectively. The Standard Bacterial Colony Index was used to assess handwashing effectiveness. RESULTS: At 6-week follow-up, performance on the Knowledge with Standard Precautions Questionnaire, Attitude with Standard Precautions Scale, and Compliance with Standard Precautions Scale were significantly improved in the intervention group compared with the control group (P < .01). The hand hygiene standard in the intervention group (38 passed) outperformed the control group (23 passed) (P < .01). CONCLUSIONS: A mixed media education intervention is effective in improving knowledge, attitude, and compliance with SPs.


Recommendations for ‘responsible behaviour’ is not a sufficient policy tool in public health emergencies


title: Complete Genome Sequence of a Brazil-Type Avian coronavirus Detected in a Chicken

text: Avian coronavirus is the causative agent of infectious bronchitis in chickens, leading to multisystemic disease that might be controlled if adequate vaccine strains are used. This paper reports the first complete genome sequence of a Brazil type of this virus (27,615 nucleotides [nt]) isolated from the kidneys of a chicken.


title: Global Organization of a Positive-strand RNA Virus Genome

text: The genomes of plus-strand RNA viruses contain many regulatory sequences and structures that direct different viral processes. The traditional view of these RNA elements are as local structures present in non-coding regions. However, this view is changing due to the discovery of regulatory elements in coding regions and functional long-range intra-genomic base pairing interactions. The ∼4.8 kb long RNA genome of the tombusvirus tomato bushy stunt virus (TBSV) contains these types of structural features, including six different functional long-distance interactions. We hypothesized that to achieve these multiple interactions this viral genome must utilize a large-scale organizational strategy and, accordingly, we sought to assess the global conformation of the entire TBSV genome. Atomic force micrographs of the genome indicated a mostly condensed structure composed of interconnected protrusions extending from a central hub. This configuration was consistent with the genomic secondary structure model generated using high-throughput selective 2′-hydroxyl acylation analysed by primer extension (i.e. SHAPE), which predicted different sized RNA domains originating from a central region. Known RNA elements were identified in both domain and inter-domain regions, and novel structural features were predicted and functionally confirmed. Interestingly, only two of the six long-range interactions known to form were present in the structural model. However, for those interactions that did not form, complementary partner sequences were positioned relatively close to each other in the structure, suggesting that the secondary structure level of viral genome structure could provide a basic scaffold for the formation of different long-range interactions. The higher-order structural model for the TBSV RNA genome provides a snapshot of the complex framework that allows multiple functional components to operate in concert within a confined context.


title: Chapter 5 Expression

text: Tight control of gene expression is highly characteristic of virus replication, although the mechanisms used to achieve this vary depending on the host cell. There are considerable differences in patterns of gene expression in prokaryotic and eukaryotic cells. Control of gene expression is also achieved at a posttranscriptional level by control over translation of proteins using a number of different mechanisms. Finally, epigenetics plays a significant role in controlling virus gene expression, just as it does for cellular genes. The net results of all these interlocking processes are a complex web of interactions between the virus genome and the machinery of the host cell.


title: Complete blood count data and leukocyte expression of cytokine genes and cytokine receptor genes associated with bovine respiratory disease in calves

text: OBJECTIVE: The purpose of this study was to evaluate potential relationships between cytokine gene expression, complete blood counts (CBC) and animals that were sick or would become sick. The CBC and the transcript abundance of cytokines and their receptors expressed in leukocytes were measured from calves at two early timepoints, and again after diagnosis with bovine respiratory disease (BRD). RESULTS: Blood was collected from calves at pre-conditioning (n = 796) and weaning (n = 791) for CBC. Blood counts were also measured for the calves with BRD (n = 13), and asymptomatic calves (n = 75) after weaning. The CBC were compared for these animals at 3 time points. At diagnosis, neutrophils were higher and basophils lower in sick animals (P < 0.05). To further characterize BRD responses, transcript abundance of 84 cytokine genes were evaluated in 5 calves with BRD and 9 asymptomatic animals at all time points. There was more data for CBC than transcript abundance; hence, animal and temporary environmental correlations between CBC and transcript abundance were exploited to improve the power of the transcript abundance data. Expression of CCL16, CXCR1, CCR1 was increased in BRD positive animals compared to controls (P-corrected < 0.1). Cytokine expression data may help to provide insight into an animal’s health. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-018-3900-x) contains supplementary material, which is available to authorized users.


title: Disinfection of aircraft: Appropriate disinfectants and standard operating procedures for highly infectious diseases

text: For infectious diseases caused by highly pathogenic agents (e. g., Ebola/Lassa fever virus, SARS-/MERS-CoV, pandemic influenza virus) which have the potential to spread over several continents within only a few days, international Health Protection Authorities have taken appropriate measures to limit the consequences of a possible spread. A crucial point in this context is the disinfection of an aircraft that had a passenger on board who is suspected of being infected with one of the mentioned diseases. Although, basic advice on hygiene and sanitation on board an aircraft is given by the World Health Organization, these guidelines lack details on available and effective substances as well as standardized operating procedures (SOP). The purpose of this paper is to give guidance on the choice of substances that were tested by a laboratory of Lufthansa Technik and found compatible with aircraft components, as well as to describe procedures which ensure a safe and efficient disinfection of civil aircrafts. This guidance and the additional SOPs are made public and are available as mentioned in this paper.


Pandémie de covid-19: rôle diagnostique fondamental de l’imagerie


Response to Letter to the Editor on “Digital Orthopaedics. A Glimpse into the Future in The Midst of a Pandemic”


Afectación endotelial por covid-19 en criobiopsia pulmonar


title: Diagnosing pneumonia in rural Thailand: Digital cameras versus film digitizers for chest radiograph teleradiology

text: Summary Background Accurate surveillance for pneumonia requires standardized classification of chest radiographs. Digital imaging permits rapid electronic transfer of data to radiologists, and recent improvements in digital camera technology present high quality, yet cheaper, options. Methods We evaluated the comparative utility of digital camera versus film digitizer in capturing chest radiographs in a pneumonia surveillance system in rural Thailand using a panel of radiologists; the gold standard was the hard-copy radiograph. We calculated sensitivity and specificity and conducted a receiver operator characteristics (ROC) analysis. Results Of the 192 radiographs from patients with clinical pneumonia, 166 (86%) were classified as pneumonia on the hard copies. Sensitivity and specificity for identifying pneumonia were 89% and 73% for the camera and 90% and 65% for the digitizer. In the ROC analysis, there was no statistically significant difference in the area under the curve (camera, 0.86; film digitizer, 0.91, p =0.29). The digital camera set cost $965 compared to $3000 for the film digitizer. Conclusion Detection of pneumonia was not measurably compromised by using digital cameras compared with film digitizers. The 3-fold lower cost of the digital camera makes this technology an affordable and widely accessible alternative for surveillance systems, vaccine trials, and perhaps clinical use.


title: Induction of lactogenic immunity to transmissible gastroneteritis virus of swine using an attenuated coronavirus mutant able to survive in the physicochemical environment of the digestive tract

text: Abstract A trasmissible gastroenteritis (TGE) coronavirus mutant (188-SG), selected as attenuated and resistant to acidity and proteases of the digestive tract of adult pigs, was used as vaccine (“Nouzilly strain”) in sows to protect suckling piglets against a challenge exposure carried out with a higly virulent TGEV strain. The pregnant sows were immunized once (42–49 days before farrowing) or twice (42–49 and 7–15 days before farrowing) by the oral, intramuscular or conjunctival route with the 188-SG strain. Sows exposed to virulent TGEV in the field and experimentally infected sows (two oral inoculations during pregnancy) were used as positive controls leading to high protection. The neutralizing antibody response to vaccination and/or infection was studied in serum and milk. No protection against mortality was observed in the litters of (1) the nine seronegative, susceptible sows, with piglet mortality of 65/70, (2) the seven once orally vaccinated sows, with mortality of 44/54, (3) the seven sows vaccinated twice by the conjunctival route, with mortality of 55/76. Moderate protection was observed in (1) the eight sows vaccinated intramuscularly twice with piglet mortality of 36/90, (2) the seven orally and intramuscularly vaccinated sows with piglet mortality of 31/51. In contrast, improved protection was observed in (1) the 10 sows vaccinated twice orally, with piglet mortality of 23/95, (2) the four naturally infected sows with piglet mortality of 6/41, (3) the six sows experimentally infected with virulent TGEV with piglet mortality of 1/59. No correlation was found between neutralizing antibodies titers in serum and milk and protection rate of the piglets. The results indicate that relative protective lactogenic immunity against TGEV is induced only by repeated ingestion of -SG attenuated 188-SG strain of TGEV.


title: Activity‐based probes for the ubiquitin conjugation–deconjugation machinery: new chemistries, new tools, and new insights

text: The reversible post‐translational modification of proteins by ubiquitin and ubiquitin‐like proteins regulates almost all cellular processes, by affecting protein degradation, localization, and complex formation. Deubiquitinases (DUBs) are proteases that remove ubiquitin modifications or cleave ubiquitin chains. Most DUBs are cysteine proteases, which makes them well suited for study by activity‐based probes. These DUB probes report on deubiquitinase activity by reacting covalently with the active site in an enzyme‐catalyzed manner. They have proven to be important tools to study DUB selectivity and proteolytic activity in different settings, to identify novel DUBs, and to characterize deubiquitinase inhibitors. Inspired by the efficacy of activity‐based probes for DUBs, several groups have recently reported probes for the ubiquitin conjugation machinery (E1, E2, and E3 enzymes). Many of these enzymes, while not proteases, also posses active site cysteine residues and can be targeted by covalent probes. In this review, we will discuss how features of the probe (cysteine‐reactive group, recognition element, and reporter tag) affect reactivity and suitability for certain experimental applications. We will also review the diverse applications of the current probes, and discuss the need for new probe types to study emerging aspects of ubiquitin biology.


title: Prior Virus Exposure Alters the Long-Term Landscape of Viral Replication during Feline Lentiviral Infection

text: We developed a feline model of lentiviral cross-species transmission using a puma lentivirus (PLV or FIV(Pco)) which infects domestic cats but does not cause disease. Infection with PLV protects cats from CD4+ T-cell decline caused by subsequent infection with virulent feline immunodeficiency virus (FIV). Previous studies implicate innate immune and/or cellular restriction mechanisms for FIV disease attenuation in PLV-infected cats. In this study, we evaluated viral infection and cytokine mRNA transcription in 12 different tissue reservoirs approximately five months post infection. We quantitated tissue proviral load, viral mRNA load and relative transcription of IL-10, IL-12p40 and IFNγ from tissues of cats exposed to FIV, PLV or both viruses and analyzed these parameters using a multivariate statistical approach. The distribution and intensity of FIV infection and IFNγ transcription differed between single and co-infected cats, characterized by higher FIV proviral loads and IFNγ expression in co-infected cat tissues. Variability in FIV mRNA load and IFNγ was significantly more constrained in co-infected versus singly infected cat tissues. Single-infected:co-infected ratios of FIV mRNA load compared to FIV proviral load indicated that active viral transcription was apparently inhibited during co-infection. These results indicate that previous PLV infection increases activation of tissue innate immunity and constrains the ability of FIV to productively infect tissue reservoirs of infection for months, independent of FIV proviral load, supporting a model in which innate immunity and/or modulation of target cell susceptibility play a key role in PLV-induced protection from FIV disease.


title: Multiple simultaneous viral infections in infants with acute respiratory tract infections in Spain

text: Abstract Background The clinical significance of the presence of more than one type of virus in the respiratory specimens of children with respiratory infections is not clear. Objectives To describe the clinical characteristics of multiple viral infections versus single infection by respiratory syncytial virus (RSV) in hospitalized infants. Study design This is a prospective study conducted in all infants under 2 years of age admitted for acute respiratory infection (September 2000–June 2003) in a secondary teaching hospital. Virological diagnosis was made by two different multiplex reverse transcription-nested polymerase chain reaction (RT-PCR) assays in nasopharyngeal aspirates. We describe the clinical characteristics of the patients with multiple viral infections and compare them to a group of 86 randomly selected patients infected only with RSV. Results 749 specimens taken were analyzed. Respiratory viruses were detected in 65.9% of the samples. 86 children had multiple viral infections (17.4% of all positive specimens). The most frequent clinical diagnosis in this group was recurrent wheezing in 44% and bronchiolitis in 52%. Fever was significantly more frequent (p <0.001), hospital stays were longer (p =0.05), and antibiotic treatment was used more (p =0.03) in infants with multiple viral infections than in the RSV-infected group. Conclusions Multiple viral infections are frequent in hospitalized children with respiratory tract disease (17.4%). Multiple viral infections are linked to higher fever, longer hospital stays and more frequent use of antibiotics than in the case of infants with single RSV infections.


title: Coronavirus (COVID-19) in Italy: knowledge, management of patients and clinical experience of Italian dentists during the spread of contagion

text: BACKGROUND: The coronavirus infection that emerged in China in the last few months of 2019 has now spread globally. Italy registered its first case in the second half of February, and in a short time period, it became the top country in Europe in terms of the number of infected people and the first in the world in terms of deaths. The medical and scientific community has been called upon to manage the emergency and to take measures. Dentists also need to take new precautions during their clinical activity to protect themselves, coworkers and patients from the risks of contagion and to avoid further spread of infection. METHODS: Following the data published in the international literature as well as the guidelines and directives constantly updated by the WHO and by the national health authorities, a questionnaire to be completed anonymously was submitted online to Italian dentists using social tools and online professional platforms. The collected data were processed statistically, providing descriptive data and analysis of correlations of the most significant parameters using the Pearson’s χ2, the Likelihood-Ratio χ2, Cramér’s V, Fisher’s exact test, Goodman and Kruskal’s γ, and Kendall’s τb (p < 0.05). RESULTS: A total of 535 dentists from Italy participated in the survey. A good level of scientific knowledge about coronavirus and the extra precautionary measures needed to limit the spread was related to the age of respondents and their sex. Coming from areas with higher concentrations of cases affected knowledge, level of attention and perception of risk related to dental activity. CONCLUSIONS: At the moment, there are no therapies or vaccines to contain the infection with the new coronavirus that is causing many infections, many of which are fatal, worldwide. Dentists are one of the categories at highest risk of encountering diseases and infections because they work in close proximity with patients, and in their procedures, there is always contact with aerosols with high bacterial and viral potential. Therefore, during this COVID-19 emergency, it is important that dentists are properly informed and take the appropriate precautionary measures.


title: Developing an ultra-efficient microsatellite discoverer to find structural differences between SARS-CoV-1 and Covid-19

text: MOTIVATION: Recently, the outbreak of Coronavirus-Covid-19 has forced the World Health Organization to declare a pandemic status. A genome sequence is the core of this virus which interferes with the normal activities of its counterparts within humans. Analysis of its genome may provide clues toward the proper treatment of patients and the design of new drugs and vaccines. Microsatellites are composed of short genome subsequences which are successively repeated many times in the same direction. They are highly variable in terms of their building blocks, number of repeats, and their locations in the genome sequences. This mutability property has been the source of many diseases. Usually the host genome is analyzed to diagnose possible diseases in the victim. In this research, the focus is concentrated on the attacker's genome for discovery of its malicious properties. RESULTS: The focus of this research is the microsatellites of both SARS and Covid-19. An accurate and highly efficient computer method for identifying all microsatellites in the genome sequences is discovered and implemented, and it is used to find all microsatellites in the Coronavirus-Covid-19 and SARS2003. The Microsatellite discovery is based on an efficient indexing technique called K-Mer Hash Indexing. The method is called Fast Microsatellite Discovery (FMSD) and it is used for both SARS and Covid-19. A table composed of all microsatellites is reported. There are many differences between SARS and Covid-19, but there is an outstanding difference which requires further investigation. AVAILABILITY: FMSD is freely available at https://gitlab.com/FUM_HPCLab/fmsd_project, implemented in C on Linux-Ubuntu system. Software related contact: hossein_savari@mail.um.ac.ir.


title: Mutational analysis of a helicase motif-based RNA 5′-triphosphatase/NTPase from bamboo mosaic virus

text: Abstract The helicase-like domain of BaMV replicase possesses NTPase and RNA 5′-triphosphatase activities. In this study, mutational effects of the helicase signature motifs and residue L543 on the two activities were investigated. Either activity was inactivated by K643A-S644A, D702A, D730A, R855A, or L543P mutations. On the other hand, Q826A, D858A and L543A had activities, in terms of k cat/K m, reduced by 5- to 15-fold. AMPPNP, a nonhydrolyzable ATP analogue, competitively inhibited RNA 5′-triphosphatase activity. Analogies of mutational effects on the two activities and approximation of K i(AMPPNP) and K m(ATP) suggest that the catalytic sites of the activities are overlapped. Mutational effects on the viral accumulation in Chenopodium quinoa indicated that the activities manifested by the domain are required for BaMV survival. Results also suggest that Q826 in motif V plays an additional role in preventing tight binding to ATP, which would otherwise decrease further RNA 5′-triphosphatase, leading to demise of the virus in plant.


title: Helicase of Type 2 Porcine Reproductive and Respiratory Syndrome Virus Strain HV Reveals a Unique Structure

text: Porcine reproductive and respiratory syndrome virus (PRRSV) is prevalent throughout the world and has caused great economic losses to the swine industry. Nonstructural protein 10 (nsp10) is a superfamily 1 helicase participating in multiple processes of virus replication and one of the three most conserved proteins in nidoviruses. Here we report three high resolution crystal structures of highly pathogenic PRRSV nsp10. PRRSV nsp10 has multiple domains, including an N-terminal zinc-binding domain (ZBD), a β-barrel domain, a helicase core with two RecA-like domains, and a C-terminal domain (CTD). The CTD adopts a novel fold and is required for the overall structure and enzymatic activities. Although each domain except the CTD aligns well with its homologs, PRRSV nsp10 adopts an unexpected extended overall structure in crystals and solution. Moreover, structural and functional analyses of PRRSV nsp10 versus its closest homolog, equine arteritis virus nsp10, suggest that DNA binding might induce a profound conformational change of PRRSV nsp10 to exert functions, thus shedding light on the mechanisms of activity regulation of this helicase.


Transmission of SARS-CoV-2 and Other Infections at Large Sports Gatherings: A Surprising Gap in Our Knowledge


title: Global Community Child Health

text: This special issue of IJERPH has published a range of studies in this developing field of Global Community Child Health research. A number of manuscripts submitted in response to our invitation describing ‘community-based interventions which impact on child health and wellbeing around the globe. In addition to rural community-based initiatives given that most children now live in cities we are also interested to hear about urban initiatives….’ We hope this issue will of great interest to the researchers and practitioners as well as academia from the fields of Global Health as well as Global Child Health because it comprised of 14 articles representing all five continents. Physical activity appears a key component of the scientific community’s current conception of child well-being judging from the four papers published addressing this area. This issue also has papers on childhood obesity to rubella vaccination. Despite of the journal’s strive for reaching out to a wider global child health community, this issue missed contributions relating to child safeguarding and social determinants of urban health.


title: Establishment of different plasmid only-based reverse genetics systems for the recovery of African horse sickness virus

text: Abstract In an effort to simplify and expand the utility of African horse sickness virus (AHSV) reverse genetics, different plasmid-based reverse genetics systems were developed. Plasmids containing cDNAs corresponding to each of the full-length double-stranded RNA genome segments of AHSV-4 under control of a T7 RNA polymerase promoter were co-transfected in cells expressing T7 RNA polymerase, and infectious AHSV-4 was recovered. This reverse genetics system was improved by reducing the required plasmids from 10 to five and resulted in enhanced virus recovery. Subsequently, a T7 RNA polymerase expression cassette was incorporated into one of the AHSV-4 rescue plasmids. This modified 5-plasmid set enabled virus recovery in BSR or L929 cells, thus offering the possibility to generate AHSV-4 in any cell line. Moreover, mutant and cross-serotype reassortant viruses were recovered. These plasmid DNA-based reverse genetics systems thus offer new possibilities for investigating AHSV biology and development of designer AHSV vaccine strains.


title: Eosinophils

text: Abstract Eosinophils have been traditionally understood as end-stage, primarily cytotoxic effector cells. Recent studies have had profound impact on this limited view and have led to new research on the functions and capabilities of this unique leukocyte lineage. Novel insights into eosinophil development, localization, modes of degranulation, and the nature of their granule contents have provided a better understanding of these cells as immunomodulatory mediators in health and disease.


title: CaMK IV phosphorylates prohibitin 2 and regulates prohibitin 2-mediated repression of MEF2 transcription

text: Prohibitin 2 (PHB2) is an evolutionarily conserved and ubiquitously expressed multifunctional protein which is present in various cellular compartments including the nucleus. However, mechanisms underlying various functions of PHB2 are not fully explored yet. Previously we showed that PHB2 interacts with Akt and inhibits muscle differentiation by repressing the transcriptional activity of both MyoD and MEF2. Here we show that Calcium/Calmodulin-dependent kinase IV (CaMK IV) specifically binds to the C terminus of PHB2 and phosphorylates PHB2 at serine 91. Ectopic expression of CaMK IV and PHB2 in C2C12 cells results effectively in decreased PHB2-mediated repression of MEF2-dependent gene expression. Conversely, PHB2 mutant (S91A) resistant to CaMK IV phosphorylation has less effective in relieving the inhibition of MEF2 transcription by PHB2. Our findings suggest that CaMK IV interacts with and regulates PHB2 through phosphorylation, which could be one of the mechanisms underlying the CaMK-mediated activation of MEF2.


title: Remote Monitoring of Oxygen Saturation in Individuals with COVID-19 Pneumonia

text: Remote monitoring of oxygen saturation in cases of COVID-19 pneumonia may facilitate discharge relieving burden on bed demand and allowing safe follow up for this disease in which the sequelae are unknown.


title: Genetic analysis of picornaviruses

text: During the past year, genetic studies of picornaviruses, vastly facilitated by the application of infectious picornaviral cDNAs and RNAs, have contributed to our understanding of the function of individual picornavirus polypeptides and to the genetic processes that operate in these small RNA viruses. Especially notable were the demonstrations that the RNA-dependent RNA polymerase may have a function in RNA synthesis as an uncleaved precursor polypeptide, and that a mutation in the polymerase can be complemented in trans, in contrast to data obtained from previously studied polymerase mutants. A new in vitro system, in which positive-strand sythesis, negative-strand synthesis and RNA packaging were all observed, will facilitate further studies into the mechanism of these processes.


title: Global site-specific analysis of glycoprotein N-glycan processing

text: N-glycans contribute to the folding, stability and functions of the proteins they decorate. They are produced by transfer of the glycan precursor to the sequon Asn-X-Thr/Ser, followed by enzymatic trimming to a high-mannose-type core and sequential addition of monosaccharides to generate complex-type and hybrid glycans. This process, mediated by the concerted action of multiple enzymes, produces a mixture of related glycoforms at each glycosite, making analysis of glycosylation difficult. To address this analytical challenge, we developed a robust semiquantitative mass spectrometry (MS)-based method that determines the degree of glycan occupancy at each glycosite and the proportion of N-glycans processed from high-mannose type to complex type. It is applicable to virtually any glycoprotein, and a complete analysis can be conducted with 30 μg of protein. Here, we provide a detailed description of the method that includes procedures for (i) proteolytic digestion of glycoprotein(s) with specific and nonspecific proteases; (ii) denaturation of proteases by heating; (iii) sequential treatment of the glycopeptide mixture with two endoglycosidases, Endo H and PNGase F, to create unique mass signatures for the three glycosylation states; (iv) LC-MS/MS analysis; and (v) data analysis for identification and quantitation of peptides for the three glycosylation states. Full coverage of site-specific glycosylation of glycoproteins is achieved, with up to thousands of high-confidence spectra hits for each glycosite. The protocol can be performed by an experienced technician or student/postdoc with basic skills for proteomics experiments and takes ∼7 d to complete. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nprot.2018.024) contains supplementary material, which is available to authorized users.


title: Identification of bioactive compounds from Glycyrrhiza glabra as possible inhibitor of SARS-CoV-2 spike glycoprotein and non-structural protein-15: a pharmacoinformatics study

text: At present, the world is facing a pandemic named as COVID-19, caused by SARS-CoV-2. Traditional Chinese medicine has recommended the use of liquorice (Glycyrrhiza species) in the treatment of infections caused by SARS-CoV-2. Therefore, the present investigation was carried out to identify the active molecule from the liquorice against different protein targets of COVID-19 using an in-silico approach. The molecular docking simulation study of 20 compounds along with two standard antiviral drugs (Lopinavir and Rivabirin) was carried out with the help of Autodock vina software using two protein targets from COVID-19 i.e. spike glycoprotein (PDB ID: 6VSB) and Non-structural Protein-15 (Nsp15) endoribonuclease (PDB ID: 6W01). From the observed binding energy and the binding interactions, glyasperin A showed high affinity towards Nsp15 endoribonuclease with uridine specificity, while glycyrrhizic acid was found to be best suited for the binding pocket of spike glycoprotein and also prohibited the entry of the virus into the host cell. Further, the dynamic behavior of the best-docked molecules inside the spike glycoprotein and Nsp15 endoribonuclease were explored through all-atoms molecular dynamics (MD) simulation study. Several parameters from the MD simulation have substantiated the stability of protein-ligand stability. The binding free energy of both glyasperin A and glycyrrhizic acid was calculated from the entire MD simulation trajectory through the MM-PBSA approach and found to high binding affinity towards the respective protein receptor cavity. Thus, glyasperin A and glycyrrhizic acid could be considered as the best molecule from liquorice, which could find useful against COVID-19. Communicated by Ramaswamy H. Sarma


title: Detection of virus-neutralising antibodies and associated factors against rabies in the vaccinated household dogs of Kathmandu Valley, Nepal

text: BACKGROUND: Rabies is a vaccine-preventable neglected tropical viral zoonosis. It occurs worldwide, creating a very heavy burden in many developing countries, including Nepal. Dogs are the principle vector for the transmission of this disease in urban areas. Vaccination is the most important preventive measure in areas where dogs are the principle source of infection. This study was conducted with the aim of detecting virus-neutralising antibodies and associated factors against rabies in vaccinated household dogs of Kathmandu valley. METHODS: Blood samples were collected from 110 vaccinated pet dogs in Kathmandu, Bhaktapur, and Lalitpur districts of Nepal. The samples were taken to the laboratory of the National Zoonosis and Food Hygiene Research Center where serum was separated. An indirect immune-enzymatic assay (Platelia(TM) Rabies II kit ad usum Veterinarium, Biorad, China) was used for the detection of rabies virus anti-glycoprotein antibodies in the dog serum samples following the manufacturer’s recommendations and instructions. Optical density values for unknown samples were compared with the positive sera titers in quantification tests obtained after a direct reading on the standard curve. Results were expressed as equivalent units per ml (EU/ml). FINDINGS: Of the total samples, 89.09% exceeded the required seroconversion level (≥ 0.5 EU/ml); another 9.09% did not reach the seroconversion level (0.125–0.5 EU/ml); and 1.81% had undetectable seroconversion levels (<0.125 EU/ml) suggesting that the animal had not seroconverted according to the PLATELIA(™) RABIES II test. Only one factor, the condition under which the dog was kept, was significantly associated with the antibody titer level. No association was found for any of the other factors included in the study. INTERPRETATION: Vaccination is the most effective measure for prevention and control of rabies. The locally manufactured brand of vaccine, which is available in Nepal, is potent enough to generate a sufficient amount of protective antibodies, equal to international brands.


title: Remdesivir is a direct-acting antiviral that inhibits RNA-dependent RNA polymerase from severe acute respiratory syndrome coronavirus 2 with high potency

text: Effective treatments for coronavirus disease 2019 (COVID-19) are urgently needed to control this current pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Replication of SARS-CoV-2 depends on the viral RNA-dependent RNA polymerase (RdRp), which is the likely target of the investigational nucleotide analogue remdesivir (RDV). RDV shows broad-spectrum antiviral activity against RNA viruses, and previous studies with RdRps from Ebola virus and Middle East respiratory syndrome coronavirus (MERS-CoV) have revealed that delayed chain termination is RDV's plausible mechanism of action. Here, we expressed and purified active SARS-CoV-2 RdRp composed of the nonstructural proteins nsp8 and nsp12. Enzyme kinetics indicated that this RdRp efficiently incorporates the active triphosphate form of RDV (RDV-TP) into RNA. Incorporation of RDV-TP at position i caused termination of RNA synthesis at position i+3. We obtained almost identical results with SARS-CoV, MERS-CoV, and SARS-CoV-2 RdRps. A unique property of RDV-TP is its high selectivity over incorporation of its natural nucleotide counterpart ATP. In this regard, the triphosphate forms of 2′-C-methylated compounds, including sofosbuvir, approved for the management of hepatitis C virus infection, and the broad-acting antivirals favipiravir and ribavirin, exhibited significant deficits. Furthermore, we provide evidence for the target specificity of RDV, as RDV-TP was less efficiently incorporated by the distantly related Lassa virus RdRp, and termination of RNA synthesis was not observed. These results collectively provide a unifying, refined mechanism of RDV-mediated RNA synthesis inhibition in coronaviruses and define this nucleotide analogue as a direct-acting antiviral.


title: Diseased prey predator model with general Holling type interactions

text: Abstract Choice of interaction function is one of the most important parts for modelling a food chain. Many models have been proposed as a diseased-prey predator model with Holling type-I or type-II or type-III interactions, but there is no model with general Holling type interactions. In this paper, we study a diseased prey–predator model with general Holling type interactions. Local stability conditions of equilibrium points are derived. We obtain the permanence and impermanence conditions of the system. The conditions for global stability of the system are also derived. The system exhibits limit cycle, period-2, higher periodic oscillations and chaotic behaviour for different values of Holling parameters. One parameter bifurcation analysis is done with respect to general Holling parameters and infection rate. We utilize the MATCONT package to analyse the detailed bifurcation scenario as the two important interaction parameters are varied. It is interesting to note that a diseased system becomes a disease free system for proper choice of interaction functions. Our results give an idea for constructing a realistic food chain model through proper choice of general Holling parameters.


Mortality in Older Patients with Covid‐19


title: Impact of mental health and caregiver burden on family caregivers’ physical health

text: Abstract Caregivers providing care to chronically ill family members at home are potentially at risk for caregiver burden and declining physical and psychological health. This study aims to understand how family caregivers’ mental health and caregiver burden affects physical health simultaneously, controlling for factors such as age, education level, caring hours per day, and emotional, functional, and physical support systems used by caregivers. We recruited 388 caregivers from Kaohsiung and Pingtong region in Taiwan. Caregivers had to be 18 years or older and spend most of their time taking care of an ill family member at home. Mental health was assessed by the 12-item Chinese Health Questionnaire (CHQ-12) and burden was measured using a modified scale for caregiver's burden. Health status was assessed by self-perceived health (SPH), illness symptoms and the number of diagnosed chronic diseases. A high number of hours per day of caregiving was associated with low emotional support and SPH, poor mental health and high burden. Higher emotional support was associated with better mental health and fewer illness symptoms. Higher physical support was associated with poorer mental health, higher burden, a greater number of illness symptoms and chronic diseases, and a lower SPH score. Hours per day of caregiving, and use of emotional, functional, and physical support were associated with mental health, and the hours per day of caregiving and use of physical support were predictors of burden. Mental health and burden were significantly associated with caregivers’ health problems simultaneously. Our results show the important influence of mental health on caregivers’ physical health. Interventions for caregivers need to target health in a holistic way.


A new diagnostic strategy for gestational diabetes during the COVID‐19 pandemic for the Japanese population