title: Personal Cleanliness Activities in Preschool Classrooms

text: One hundred and twelve preschool teachers in Indiana were asked to complete a questionnaire requesting them to indicate what personal health-related activities they were incorporating into their classroom routines, why they were incorporating them, and how often such activities were done. The results of the study indicate that the most incorporated activities were hand-washing, proper ways of coughing, and discussion on germs. The least incorporated personal health-related activities were proper use of bathroom and keeping things out of the mouth. Reasons for incorporating the activities include helping to curtail the spreading of colds and germs, helping the children to develop good life skills later in life, and helping the preschoolers know about the health hazards associated with germs, among others. The study recommends the need to make Health Education a core component of preschool teachers’ training.


title: Hospital Preparedness and SARS

text: On May 23, 2003, Toronto experienced the second phase of a severe acute respiratory syndrome (SARS) outbreak. Ninety cases were confirmed, and >620 potential cases were managed. More than 9,000 persons had contact with confirmed or potential case-patients; many required quarantine. The main hospital involved during the second outbreak was North York General Hospital. We review this hospital’s response to, and management of, this outbreak, including such factors as building preparation and engineering, personnel, departmental workload, policies and documentation, infection control, personal protective equipment, training and education, public health, management and administration, follow-up of SARS patients, and psychological and psychosocial management and research. We also make recommendations for other institutions to prepare for future outbreaks, regardless of their origin.


docid: otyrbdwq

title: Addressing problems with alcohol and other substances among older adults during the COVID-19 pandemic


title: Basic Scholarship in Biosafety Is Critically Needed To Reduce Risk of Laboratory Accidents

text: Our firm conducted a risk/benefit assessment of “gain-of-function” research, as part of the deliberative process following a U.S. moratorium on the research (U.S. Department of Health and Human Services, U.S. Government Gain-of-Function Deliberative Process and Research Funding Pause on Selected Gain-of-Function Research Involving Influenza, MERS, and SARS Viruses, 2014). Due to significant missing but theoretically acquirable data, our biosafety assessment faced limitations, and we were forced to provide a relative, instead of absolute, measure of risk (Gryphon Scientific, LLC, Risk and Benefit Analysis of Gain of Function Research, 2016). Here, we argue that many of these types of missing data represent large and stunning gaps in our knowledge of biosafety and argue that these missing data, once acquired via primary research efforts, would improve biosafety risk assessments and could be incorporated into biosafety practices to reduce risk of accidents. Governments invest billions in biological research; at least a small fraction of this support is warranted to prevent biological accidents.


title: Unexpected rise of ozone in urban and rural areas, and sulfur dioxide in rural areas during the coronavirus city lockdown in Hangzhou, China: implications for air quality

text: The outbreak of coronavirus named COVID-19, initially identified in Wuhan, China in December 2019, has spread rapidly at the global scale. Most countries have rapidly stopped almost all activities including industry, services and transportation of goods and people, thus decreasing air pollution in an unprecedented way, and providing a unique opportunity to study air pollutants. While satellite data have provided visual evidence for the global reduction in air pollution such as nitrogen dioxide (NO(2)) worldwide, precise and quantitative information is missing at the local scale. Here we studied changes in particulate matter (PM(2.5), PM(10)), carbon monoxide (CO), NO(2), sulfur dioxide (SO(2)) and ozone (O(3)) at 10 urban sites in Hangzhou, a city of 7.03 million inhabitants, and at 1 rural site, before city lockdown, January 1–23, during city lockdown, January 24-February 15, and during resumption, February 16–28, in 2020. Results show that city lockdown induced a sharp decrease in PM(2.5), PM(10), CO, and NO(2) concentrations at both urban and rural sites. The NO(2) decrease is explained by reduction in traffic emissions in the urban areas, and by lower regional transport in rural areas during lockdown, as expected. SO(2) concentrations decreased from 6.3 to 5.3 μg m(−3) in the city, but increased surprisingly from 4.7 to 5.8 μg m(−3) at the rural site: this increase is attributed both to higher coal consumption for heating and emissions from traditional fireworks of the Spring Eve and Lantern Festivals during lockdown. Unexpectedly, O(3) concentrations increased by 145% from 24.6 to 60.6 μg m(−3) in the urban area, and from 42.0 to 62.9 μg m(−3) in the rural area during the lockdown. This finding is explained by the weakening of chemical titration of O(3) by NO due to reductions of NO(x) fresh emissions during the non-photochemical reaction period from 20:00 PM to 9:00 AM (local time). During the lockdown, compared to the same period in 2019, the daily average concentrations in the city decreased by 42.7% for PM(2.5), 47.9% for PM(10), 28.6% for SO(2), 22.3% for CO and 58.4% for NO(2), which is obviously explained by the absence of city activities. Overall, we observed not only the expected reduction in some atmospheric pollutants (PM, SO(2), CO, NO(2)), but also unexpected increases in SO(2) in the rural areas and of ozone (O(3)) in both urban and rural areas, the latter being paradoxically due to the reduction in nitrogen oxide levels. In other words, the city lockdown has improved air quality by reducing PM(2.5), PM(10), CO, and NO(2), but has also decreased air quality by augmenting O(3) and SO(2). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10311-020-01028-3) contains supplementary material, which is available to authorized users.


title: A DESCRIPTIVE STUDY OF PANDEMIC INFLUENZA A(H1N1)PDM09 IN BRAZIL, 2009 - 2010

text: Influenza A viruses undergo frequent antigenic mutations and may thus cause seasonal epidemics and pandemics. The aim of this study was to recover the epidemiological history of the pandemic influenza A(H1N1)pdm09 in Brazil. A descriptive study was conducted in 2009-2010. The Brazilian Information System for reportable diseases (SINAN) was the data source. A total of 105,054 suspected cases of influenza A(H1N1)pdm09 were reported to SINAN. Of these, 53,797 (51.2%) were classified as the new influenza virus subtype. Among the confirmed cases, 56.7% were female, the mean age was 26.31 (SD ± 18.1) years. Fever was the most common sign among the confirmed cases (99.7%) and the presence of comorbidities was reported in 32.5% of cases. In 2009 there were confirmed cases in all 26 Brazilian States and the Federal District. The incidence (per 100,000 inhabitants) of severe influenza in the population was 28.0 in 2009 and 0.5 in 2010. The states of Paraná (301.3), Santa Catarina (36.0) and Rio Grande do Sul (27.4) presented the highest incidence; 46.4% of the confirmed cases were hospitalized and 47,643 were cured (93.8%). The case-fatality rate was 3.9% in 2009. The pandemic virus A(H1N1)pdm09 hit Brazil between April/2009 and December/2010 with an important difference in the geographic pattern distribution of the cases from the northeast to the south of the country. Children and young adults were the most affected. The limitations of the study were data quality and inconsistencies in the final classification of cases in SINAN. This study highlights the urgent need for improvements in the surveillance of emerging diseases in Brazil.


title: The human coronavirus HCoV-229E S-protein structure and receptor binding

text: The coronavirus S-protein mediates receptor binding and fusion of the viral and host cell membranes. In HCoV-229E, its receptor binding domain (RBD) shows extensive sequence variation but how S-protein function is maintained is not understood. Reported are the X-ray crystal structures of Class III-V RBDs in complex with human aminopeptidase N (hAPN), as well as the electron cryomicroscopy structure of the 229E S-protein. The structures show that common core interactions define the specificity for hAPN and that the peripheral RBD sequence variation is accommodated by loop plasticity. The results provide insight into immune evasion and the cross-species transmission of 229E and related coronaviruses. We also find that the 229E S-protein can expose a portion of its helical core to solvent. This is undoubtedly facilitated by hydrophilic subunit interfaces that we show are conserved among coronaviruses. These interfaces likely play a role in the S-protein conformational changes associated with membrane fusion.


title: Safety and Immunogenicity of High Dose Trivalent Inactivated Influenza Vaccine in Pediatric Patients with Acute Lymphoblastic Leukemia

text: BACKGROUND: Although children with acute lymphoblastic leukemia (ALL) mount immune responses after vaccination with the trivalent influenza vaccine (TIV), these responses are lower compared to controls. Recently, a high dose (HD) TIV was found to increase the level of antibody response in elderly patients compared to the standard dose (SD) TIV. We hypothesized that the HD TIV would be well-tolerated and more immunogenic compared to the SD TIV in pediatric subjects with ALL. PROCEDURE: This was a randomized, double-blind, phase I safety and immunogenicity trial comparing the HD to the SD TIV in children with ALL. Subjects were randomized 2:1 to receive either the HD (60µg) or the SD (15µg) TIV. Local and systemic reactions were solicited, hemagglutinin inhibition titers to influenza virus antigens were measured, and monitoring labs were collected prior to and/or after each vaccination. RESULTS: Fifty subjects were enrolled (34 HD, 16 SD). Mean age was 8.5 years; 63% were male, and 80% were in maintenance therapy. There were no significant differences reported in local or systemic symptoms. No severe adverse events were attributed to vaccination. No significant differences between the HD and SD TIV groups were noted for immune responses. CONCLUSIONS: No differences were noted between the HD and SD TIV groups for solicited systemic and local reactions. Since this study was not powered for immunogenicity, a phase II trial is needed to determine the immunogenicity of HD versus SD TIV in the pediatric ALL population.


title: Chapter 84 Respiratory Tract Infections A Clinical Approach

text: Respiratory tract infections are among the most common and important problems in clinical medicine. The approach to these infections begins with a basic understanding of the pathophysiology of the respiratory tract and how immune defences interact with pathogens. A number of distinct clinical syndromes can be defined which help physicians and clinical microbiologists develop a differential diagnosis of potential infecting pathogens. The type and severity of infection is often modulated by the medical characteristics of the patient, circumstances surrounding infection, geography and environmental exposures. Multiple laboratory test methodologies are available to aid clinical diagnosis and close communication between clinicians and laboratory personnel can optimise the efficiency and cost-effectiveness of diagnosis. Molecular testing for respiratory viruses has become widely used in the clinical setting and will expand to include other pathogens of the respiratory tract.


title: Evidence of MAPK–JNK1/2 activation by hepatitis E virus ORF3 protein in cultured hepatoma cells

text: Hepatitis E virus (HEV) has recently emerged to cause chronic infection in some immunosuppressed individuals, including extrahepatic manifestations in acute and chronic patients. Mammalian MAPK–JNK1/2 is expressed in hepatocytes, which is known to be involved in anti-apoptotic signaling pathway for the establishment of persistent infection. Though in vitro modulation of cellular MAPK–ERK cascade by HEV-ORF3 protein is suggested to have a role in host pathobiology, activation of the JNK module has not been studied so far. In this report, we have shown for the first time, evidence of MAPK–JNK1/2 activation by HEV-ORF3, using viral replicon as well as expression vector in human hepatoma cells. Phospho-ELISA based relative quantitaion has demonstrated ~54% and ~66% phosphorylation of JNK1/2 in replicon-RNA and ORF3-vector DNA transfected cells, respectively. Our finding however, suggests further molecular studies to validate a role of JNK1/2 in HEV pathogenesis.


title: The clinical data from 19 critically ill patients with coronavirus disease 2019: a single-centered, retrospective, observational study

text: BACKGROUND: The objectives of this study were to analyze the clinical features of coronavirus disease 2019 (COVID-19) and evaluate the diagnosis and treatment. METHODS: A retrospective analysis of the clinical manifestation and auxiliary examination of 19 patients with COVID-19 from the Liyuan Hospital intensive care unit (ICU) between January 16, 2020 and February 20, 2020 was undertaken. RESULTS: There were 11 male and 8 female cases among the patients. The median (range) age was 73 (38–91) years. Of these patients, 8 (42.1%) had died and the median duration from ICU admission to death was 2 (interquartile range (IQR): 1–10.75) days. Seven of these 8 patients had underlying diseases. The auxiliary examination showed fever (68.4%), dry cough (15.8%), dyspnea (10.5%), and diarrhea (5.3%). All 19 cases showed ground-glass changes on chest computed tomography. Serum hypersensitive C-reactive protein (hs-CRP) and serum amylase A (SAA) were clearly increased in all of the cases. Among the 19 cases, there were 16 (84.2%) cases in which the total number of lymphocytes decreased, 12 cases (63%) had reduced liver function, and 11 cases (58%) had deviant results for fibrinogen (FIB) and D-dimer, in particular, the D-dimer level was significantly higher in the non-survivors compared with the survivors. CONCLUSION: There were more men than women among critically ill patients. All of the cases showed ground-glass changes on chest computed tomography and the vast majority of patients displayed fever and dry cough. The clinical laboratory indices change significantly, especially the D-dimer level among non-survivors.


Restructuring Burns Management during the COVID-19 Pandemic: A Malaysian Experience


title: Broad-Spectrum Detection of H5 Subtype Influenza A Viruses with a New Fluorescent Immunochromatography System

text: Immunochromatography (IC) is an antigen-detection assay that plays an important role in the rapid diagnosis of influenza virus because the protocol is short time and easy to use. Despite the usability of IC, the sensitivity is approximately 10(3) pfu per reaction. In addition, antigen-antibody interaction-based method cannot be used for the detection of influenza viruses with major antigenic change. In this study, we established the use of fluorescent immunochromatography (FLIC) to detect a broad spectrum of H5 subtype influenza A viruses. This method has improved sensitivity 10–100 fold higher than traditional IC because of the use of fluorescent conjugated beads. Our Type-E FLIC kit detected all of the H5 subtype influenza viruses that were examined, as well as recombinant hemagglutinin (HA) proteins (rHAs) belonging to the Eurasian H5 subtype viruses and the Type-N diagnosed North American H5 subtype influenza A viruses. Thus, this kit has the improved potential to detect H5 subtype influenza viruses of different clades with both Type-E and Type-N FLIC kits. Compared with PCR-based diagnosis, FLIC has a strong advantage in usability, because the sample preparation required for FLIC is only mix-and-drop without any additional steps such as RNA extraction. Our results can provide new strategies against the spread and transmission of HPAI H5N1 viruses in birds and mammals including humans.


COVID‐19 infection on IL‐23 inhibition


title: Role of RNA Guanine Quadruplexes in Favoring the Dimerization of SARS Unique Domain in Coronaviruses

text: [Image: see text] Coronaviruses may produce severe acute respiratory syndrome (SARS). As a matter of fact, a new SARS-type virus, SARS-CoV-2, is responsible for the global pandemic in 2020 with unprecedented sanitary and economic consequences for most countries. In the present contribution we study, by all-atom equilibrium and enhanced sampling molecular dynamics simulations, the interaction between the SARS Unique Domain and RNA guanine quadruplexes, a process involved in eluding the defensive response of the host thus favoring viral infection of human cells. Our results evidence two stable binding modes involving an interaction site spanning either the protein dimer interface or only one monomer. The free energy profile unequivocally points to the dimer mode as the thermodynamically favored one. The effect of these binding modes in stabilizing the protein dimer was also assessed, being related to its biological role in assisting the SARS viruses to bypass the host protective response. This work also constitutes a first step in the possible rational design of efficient therapeutic agents aiming at perturbing the interaction between SARS Unique Domain and guanine quadruplexes, hence enhancing the host defenses against the virus.


title: Utility Evaluation of Porcine Enteroids as PDCoV Infection Model in vitro

text: Porcine deltacoronavirus (PDCoV) is a novel emerging enteric coronavirus found in pigs. Intestinal enteroids, which partially recreate the structure and function of intestinal villi-crypts, have many physiological similarities to the intestinal tissues in vivo. Enteroids exhibit advantages in studying the interactions between intestines and enteric pathogens. To create a novel infection model for PDCoV, we developed an in vitro system to generate porcine intestinal enteroids from crypts of duodenum, jejunum, and ileum of pigs. Enterocytes, enteroendocrine cells, Paneth cells, stem cells, proliferating cells, and goblet cells were found in the differentiated enteroids. Replication of PDCoV was detected in the cultured enteroids by immunofluorescence and quantitative RT-PCR. Double immunofluorescence labeling demonstrated that PDCoV was present in Sox9-positive intestinal cells and Villin1-positive enterocytes. There were multiple cellular responses shown as changes of transcription of genes related to mucosal immunity, antiviral genes, and marker genes of stem cells and other cells in the enteroids infected with PDCoV. We conclude that the 2-D enteroids derived from porcine jejunum can be used as an in vitro multicellular model for the investigation of pathogenesis and host immune responses to porcine enteric pathogens, such as PDCoV.


title: Coinfection of diarrheagenic bacterial and viral pathogens in piglets of Northeast region of India

text: AIM: This study aimed to study the prevalence of the coinfection of enteric bacterial and viral pathogens, namely Escherichia coli, Salmonella, Rotavirus, and Picobirnavirus from fecal samples of pre-weaned piglets in Northeast region of India. MATERIALS AND METHODS: A total of 457 fresh fecal samples were collected from piglets under 9 weeks old during 2013-2015 from organized (n=225) and unorganized (n=232) farms of Manipur, Meghalaya, Mizoram, and Nagaland. Samples were collected from diarrheic (n =339) and non-diarrheic (n=118) piglets including local indigenous (n=130) and crossbreed (n=327) piglets in different seasons during the study period. The samples were processed for the isolation of E. coli and Salmonella and detection of their putative virulence genes by polymerase chain reaction (PCR). Samples were also processed for the detection of Rotavirus and Picobirnavirus by RNA-polyacrylamide agarose gel electrophoresis and reverse transcriptase-PCR (RT-PCR). RESULTS: A total of 11 (2.40%) samples were found positive for two or more coinfecting enteric bacterial and viral pathogens. All the 11 positive fecal samples were recovered from diarrheic piglets. Salmonella Typhimurium (enterotoxin, stn gene) and Picobirnavirus genogroup 1 were found to be more frequent as coinfecting agents. Coinfection was recorded higher in unorganized (3.87%) compared to organized farm (0.88%). Again, higher detection was recorded in crossbreed (2.75%) than local indigenous piglets (1.53%). The occurrence of coinfection was found to be more common during summer (4.68%) followed by winter (2.27%) season. CONCLUSION: The present study highlighted the significance of E. coli, Salmonella, Rotavirus, and Picobirnavirus as important diarrheagenic pathogens causing coinfection in piglets in Northeast region of India. Probably, this is the first systematic study of the coinfection of four important diarrheagenic bacterial and viral agents associated with piglet diarrhea in India.


title: Evidence for the interaction of the human metapneumovirus G and F proteins during virus-like particle formation

text: BACKGROUND: Human metapneumovirus (HMPV) is now a major cause of lower respiratory infection in children. Although primary isolation of HMPV has been achieved in several different cell lines, the low level of virus replication and the subsequent recovery of low levels of infectious HMPV have hampered biochemical studies on the virus. These experimental methodologies usually require higher levels of biological material that can be achieved following HMPV infection. In this study we demonstrate that expression of the HMPV F, G and M proteins in mammalian cells leads to HMPV virus-like particles (VLP) formation. This experimental strategy will serve as a model system to allow the process of HMPV virus assembly to be examined. METHODS: The HMPV F, G and M proteins were expressed in mammalian cell lines. Protein cross-linking studies, sucrose gradient centrifugation and in situ imaging was used to examine interactions between the virus proteins. VLP formation was examined using sucrose density gradient centrifugation and electron microscopy analysis. RESULTS: Analysis of cells co-expressing the F, G and M proteins demonstrated that these proteins interacted. Furthermore, in cells co-expression the three HMPV proteins the formation VLPs was observed. Image analysis revealed the VLPs had a similar morphology to the filamentous virus morphology that we observed on HMPV-infected cells. The capacity of each protein to initiate VLP formation was examined using a VLP formation assay. Individual expression of each virus protein showed that the G protein was able to form VLPs in the absence of the other virus proteins. Furthermore, co-expression of the G protein with either the M or F proteins facilitated their incorporation into the VLP fraction. CONCLUSION: Co-expression of the F, G and M proteins leads to the formation of VLPs, and that incorporation of the F and M proteins into VLPs is facilitated by their interaction with the G protein. Our data suggests that the G protein plays a central role in VLP formation, and further suggests that the G protein may also play a role in the recruitment of the F and M proteins to sites of virus particle formation during HMPV infection.


title: HAb18G/CD147 cell-cell contacts confer resistance of a HEK293 subpopulation to anoikis in an E-cadherin-dependent manner

text: BACKGROUND: Acquisition of resistance to "anoikis" facilitates the survival of cells under independent matrix-deficient conditions, such as cells in tumor progression and the production of suspension culture cells for biomedical engineering. There is evidence suggesting that CD147, an adhesion molecule associated with survival of cells in tumor metastasis and cell-cell contacts, plays an important role in resistance to anoikis. However, information regarding the functions of CD147 in mediating cell-cell contacts and anoikis-resistance remains limited and even self-contradictory. RESULTS: An anoikis-resistant clone (HEK293ar), derived from anoikis-sensitive parental Human Embryonic Kidney 293 cells, survived anoikis by the formation of cell-cell contacts. The expression of HAb18G/CD147 (a member of the CD147 family) was upregulated and the protein was located at cell-cell junctions. Upregulation of HAb18G/CD147 in suspended HEK293ar cells suppressed anoikis by mediating the formation of cell-cell adhesions. Anoikis resistance in HEK293ar cells also required E-cadherin-mediated cell-cell contacts. Knock-down of HAb18G/CD147 and E-cadherin inhibited cell-cell contacts formation and increased anoikis sensitivity respectively. When HAb18G/CD147 was downregulated, E-cadherin expression in HEK293ar cells was significantly suppressed; however, knockdown of E-cadherin by E-cadherin siRNA or blocking of E-cadherin binding activity with a specific antibody and EDTA had no significant effect on HAb18G/CD147 expression. Finally, pretreatment with LY294002, a phosphoinositide 3-kinase (PI3K/AKT) inhibitor, disrupted cell-cell contacts and decreased cell number, but this was not the case in cells treated with the extracellular signal-regulated kinase (ERK) inhibitor PD98059. CONCLUSIONS: Our results provide new evidence that HAb18G/CD147-mediated cell-cell contact confers anoikis resistance in an E-cadherin-dependent manner; and cell-cell contact mediated resistance to anoikis implicates PI3K pathway in a highly relevant cell model (HEK293ar). Understanding of the role of HAb18G/CD147 cell-cell contacts in anoikis resistance may help in understanding the survival of cells in anchorage-independent growth, such as cells in tumor metastasis and suspension culture produced for biomedical engineering. Our results also contribute to a better understanding of the biology of HEK293 cell spheroids, a major workhorse for producing human therapeutic agents and viral vaccines.


title: The use of internet analytics by a Canadian provincial chiropractic regulator to monitor, evaluate and remediate misleading claims regarding specific health conditions, pregnancy, and COVID-19

text: BACKGROUND: Internet analytics are increasingly being integrated into public health regulation. One specific application is to monitor compliance of website and social media activity with respect to jurisdictional regulations. These data may then identify breaches of compliance and inform disciplinary actions. Our study aimed to evaluate the novel use of internet analytics by a Canadian chiropractic regulator to determine their registrants compliance with three regulations related to specific health conditions, pregnancy conditions and most recently, claims of improved immunity during the COVID-19 crisis. METHODS: A customized internet search tool (Market Review Tool, MRT) was used by the College of Chiropractors of British Columbia (CCBC), Canada to audit registrants websites and social media activity. The audits extracted words whose use within specific contexts is not permitted under CCBC guidelines. The MRT was first used in October of 2018 to identify words related to specific health conditions. The MRT was again used in December 2019 for words related to pregnancy and most recently in March 2020 for words related to COVID-19. In these three MRT applications, potential cases of word misuse were evaluated by the regulator who then notified the practitioner to comply with existing regulations by a specific date. The MRT was then used on that date to determine compliance. Those found to be non-compliant were referred to the regulator’s inquiry committee. We mapped this process and reported the outcomes with permission of the regulator. RESULTS: In September 2018, 250 inappropriate mentions of specific health conditions were detected from approximately 1250 registrants with 2 failing to comply. The second scan for pregnancy related terms of approximately1350 practitioners revealed 83 inappropriate mentions. Following notification, all 83 cases were compliant within the specified timeframe. Regarding COVID-19 related words, 97 inappropriate mentions of the word “immune” were detected from 1350 registrants with 7 cases of non-compliance. CONCLUSION: Internet analytics are an effective way for regulators to monitor internet activity to protect the public from misleading statements. The processes described were effective at bringing about rapid practitioner compliance. Given the increasing volume of internet activity by healthcare professionals, internet analytics are an important addition for health care regulators to protect the public they serve.


title: Contemporary Epidemiology of North American Lineage Triple Reassortant Influenza A Viruses in Pigs

text: The 2009 pandemic H1N1 infection in humans has been one of the greatest concerns for public health in recent years. However, influenza in pigs is a zoonotic viral disease well-known to virologists for almost one century with the classical H1N1 subtype the only responsible agent for swine influenza in the United States for many decades. Swine influenza was first recognized clinically in pigs in the Midwestern U.S. in 1918 and since that time it has remained important to the swine industry throughout the world. Since 1998, however, the epidemiology of swine influenza changed dramatically. A number of emerging subtypes and genotypes have become established in the U.S. swine population. The ability of multiple influenza virus lineages to infect pigs is associated with the emergence of reassortant viruses with new genomic arrangements, and the introduction of the 2009 pandemic H1N1 from humans to swine represents a well-known example. The recent epidemiological data regarding the current state of influenza A virus subtypes circulating in the Canadian and American swine population is discussed in this review.


“Time to recharge”


title: Detection of SARS‐CoV‐2 by RT‐PCR in anal from patients who have recovered from coronavirus disease 2019

text: An outbreak of coronavirus disease 2019 (COVID‐19) pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), started in Wuhan City, Hubei Province, China. The real‐time reverse‐ transcriptase polymerase ‐ chain ‐ reaction (RT ‐ PCR) method can be used for the detection of SARS‐CoV‐2 in oral swabs(1). Now, results have confirmed the presence of the live virus in stool samples from patients with COVID‐ 19(2). This article is protected by copyright. All rights reserved.


title: Acute abdomen -like-presentation associated with SARS-CoV-2 infection

text: As the cases of COVID-19 are flooding around the world, atypical presentations are being recognized, making the diagnosis challenging. Gastrointestinal symptoms and mild abdominal pain are common. However, severe abdominal pain associated with COVID-19 warranting surgical evaluation has been rarely described; recognizing such presentations and differentiating them from a surgical abdomen is critical to effectively and safely manage COVID-19 patients. Here we present a case of a middle-aged gentleman who developed features resembling secondary peritonitis. Eventually, he was found to have COVID-19 and was managed conservatively. In this report, we discuss his management course, and we explore pertinent relevant literature.


title: Supercritical Carbon Dioxide Application Using Hydantoin Acrylamide for Biocidal Functionalization of Polyester

text: Biocidal functionalization in polyester fibers is a really tough challenge because of the lack of tethering groups. This study indicated supercritical carbon dioxide application using N-halamine would be an alternative solution for obtaining antibacterial function on the polyester surface. Firstly, N-(2-methyl-1-(4-methyl-2,5-dioxo-imidazolidin-4 yl)propan-2 yl)acrylamide was synthesized and applied to the polyester in supercritical carbon dioxide medium, at 120 °C, 30 MPa for different processing times. The addition of N-halamine on the surface significantly brought antibacterial activity against E. coli. The chlorine loadings showed that 6 -h exposure time was critical to obtain sufficient antibacterial activity. This treatment caused a reasonable and tolerable loss in color and mechanical properties. But, the durability to abrasion, stability, and rechargeability of oxidative chlorine, and the durability of N-halamine on the surface were remarkably good. Conclusively, it can be available to work on polyester surfaces with resource-efficient and eco-friendly supercritical carbon dioxide technique for getting more functionalization and modification.


title: The Potential of Antimicrobial Peptides as an Antiviral Therapy against COVID-19

text: [Image: see text] COVID-19 is currently considered as a life-threatening pandemic viral infection. Finding an antiviral drug or a vaccine is the only route for humans’ survival against it. To date, no specific antiviral treatment has been confirmed. Antimicrobial peptides (AMPs) have been widely regarded as a promising solution to combat harmful microorganisms. They are biologically active molecules produced by different organisms as an essential component of their innate immune response against invading pathogens. Lactoferrin (LF), one of the AMPs, is an iron-binding glycoprotein that is present in several mucosal secretions. The antiviral activity of LF exists against a wide range of human and animal viruses (DNA and RNA). LF was proven to increase the host immunity against viral infection. Since LF is one of the constituents of breast milk and significantly located at the mucosal layers of the human body, it is considered the first line of defense against microbial infection. LF was reported to have antiviral activity against SARS-CoV infection. The significant antiviral activity of LF makes it a potential option as an immunity enhancer, a drug or a drug conjugate with conventional antivirals. The affordability, environmental safety, and efficiency of LFs will make them superior to all other control strategies.


title: COVID-19 versus seasonal influenza 2019–2020: USA

text: Severe mitigation efforts in the USA to reduce the incidence of COVID-19 infections have led to a massive rise in unemployment, social disruption, and appear to be leading to a severe economic depression. In contrast, no such interventions were regarded as necessary to manage seasonal influenza in 2019–2020. Another mitigation approach is proposed for COVID-19 that would allow society to function and yet should still be effective.


title: Outbreak of coinfection with human metapneumovirus and measles virus resulting in the death of a child at a hospital in China

text: Two children with different digestive diseases were admitted to the gastroenterology department of a children's hospital in Hangzhou, Zhejiang Province, China, in May 2010. They manifested successively acute lower respiratory tract infection symptoms during their stay in the hospital. The epidemiologic and experimental evidence supports that one child acquired nosocomial coinfection with measles virus and human metapneumovirus from another child while they shared the same ward.


title: Knowledge and Practices Regarding Safe Household Cleaning and Disinfection for COVID-19 Prevention — United States, May 2020

text: A recent report described a sharp increase in calls to poison centers related to exposures to cleaners and disinfectants since the onset of the coronavirus disease 2019 (COVID-19) pandemic (1). However, data describing cleaning and disinfection practices within household settings in the United States are limited, particularly concerning those practices intended to prevent transmission of SARS-CoV-2, the virus that causes COVID-19. To provide contextual and behavioral insight into the reported increase in poison center calls and to inform timely and relevant prevention strategies, an opt-in Internet panel survey of 502 U.S. adults was conducted in May 2020 to characterize knowledge and practices regarding household cleaning and disinfection during the COVID-19 pandemic. Knowledge gaps were identified in several areas, including safe preparation of cleaning and disinfectant solutions, use of recommended personal protective equipment when using cleaners and disinfectants, and safe storage of hand sanitizers, cleaners, and disinfectants. Thirty-nine percent of respondents reported engaging in nonrecommended high-risk practices with the intent of preventing SARS-CoV-2 transmission, such as washing food products with bleach, applying household cleaning or disinfectant products to bare skin, and intentionally inhaling or ingesting these products. Respondents who engaged in high-risk practices more frequently reported an adverse health effect that they believed was a result of using cleaners or disinfectants than did those who did not report engaging in these practices. Public messaging should continue to emphasize evidence-based, safe practices such as hand hygiene and recommended cleaning and disinfection of high-touch surfaces to prevent transmission of SARS-CoV-2 in household settings (2). Messaging should also emphasize avoidance of high-risk practices such as unsafe preparation of cleaning and disinfectant solutions, use of bleach on food products, application of household cleaning and disinfectant products to skin, and inhalation or ingestion of cleaners and disinfectants.


Risky behavior in the Ebola zone


Innate Immunity of the Lung


title: MoSAIC: Mobile Surveillance for Acute Respiratory Infections and Influenza-Like Illness in the Community

text: Surveillance for acute respiratory infection (ARI) and influenza-like illness (ILI) relies primarily on reports of medically attended illness. Community surveillance could mitigate delays in reporting, allow for timely collection of respiratory tract samples, and characterize cases of non–medically attended ILI representing substantial personal and economic burden. Text messaging could be utilized to perform longitudinal ILI surveillance in a community-based sample but has not been assessed. We recruited 161 households (789 people) in New York City for a study of mobile ARI/ILI surveillance, and selected reporters received text messages twice weekly inquiring whether anyone in the household was ill. Home visits were conducted to obtain nasal swabs from persons with ARI/ILI. Participants were primarily female, Latino, and publicly insured. During the 44-week period from December 2012 through September 2013, 11,282 text messages were sent. In responses to 8,250 (73.1%) messages, a household reported either that someone was ill or no one was ill; 88.9% of responses were received within 4 hours. Swabs were obtained for 361 of 363 reported ARI/ILI episodes. The median time from symptom onset to nasal swab was 2 days; 65.4% of samples were positive for a respiratory pathogen by reverse-transcriptase polymerase chain reaction. In summary, text messaging promoted rapid ARI/ILI reporting and specimen collection and could represent a promising approach to timely, community-based surveillance.


title: The Impact of SARS-Cov-2 Virus Infection on the Endocrine System

text: SARS-CoV-2 virus has spread across the globe rapidly causing an unprecedented pandemic. Due to the novelty of the disease, the possible impact on endocrine system is not clear. In order to compile a mini-review describing possible endocrine consequences of SARS-CoV-2 infection we performed a literature survey using the key words Covid-19, Coronavirus, SARS CoV-1, SARS Cov-2, Endocrine and related terms in medical databases including PubMed, Google Scholar, and MedARXiv from year 2000. Additional references were identified through manual screening of bibliographies and via citations in the selected articles. The literature review is current until April 28, 2020. In light of the literature, we discuss SARS-CoV-2 and explore the endocrine consequences based on the experience with structurally-similar SARS-CoV-1. Studies from the SARS -CoV-1 epidemic have reported variable changes in the endocrine organs. SARS-CoV-2 attaches to the ACE2 system in the pancreas causing perturbation of insulin production resulting in hyperglycemic emergencies. In patients with pre-existing endocrine disorders who develop COVID-19, several factors warrant management decisions. Hydrocortisone dose-adjustments are required in patients with adrenal insufficiency. Identification and management of Critical Illness Related Corticosteroid Insufficiency is crucial. Patients with Cushing’s syndrome may have poorer outcomes due to the associated immunodeficiency and coagulopathy. Vitamin D deficiency appears to be associated with increased susceptibility or severity to SARS-CoV-2 infection, and replacement may improve outcomes. Robust strategies required for the optimal management of endocrinopathies in COVID-19 are discussed extensively in this mini-review.


title: Human–Dromedary Camel Interactions and the Risk of Acquiring Zoonotic Middle East Respiratory Syndrome Coronavirus Infection

text: Middle East respiratory syndrome coronavirus (MERS‐CoV) cases without documented contact with another human MERS‐CoV case make up 61% (517/853) of all reported cases. These primary cases are of particular interest for understanding the source(s) and route(s) of transmission and for designing long‐term disease control measures. Dromedary camels are the only animal species for which there is convincing evidence that it is a host species for MERS‐CoV and hence a potential source of human infections. However, only a small proportion of the primary cases have reported contact with camels. Other possible sources and vehicles of infection include food‐borne transmission through consumption of unpasteurized camel milk and raw meat, medicinal use of camel urine and zoonotic transmission from other species. There are critical knowledge gaps around this new disease which can only be closed through traditional field epidemiological investigations and studies designed to test hypothesis regarding sources of infection and risk factors for disease. Since the 1960s, there has been a radical change in dromedary camel farming practices in the Arabian Peninsula with an intensification of the production and a concentration of the production around cities. It is possible that the recent intensification of camel herding in the Arabian Peninsula has increased the virus' reproductive number and attack rate in camel herds while the ‘urbanization’ of camel herding increased the frequency of zoonotic ‘spillover’ infections from camels to humans. It is reasonable to assume, although difficult to measure, that the sensitivity of public health surveillance to detect previously unknown diseases is lower in East Africa than in Saudi Arabia and that sporadic human cases may have gone undetected there.


Do the current cases reported to the WHO provide a realistic incidence rate of countries infected with COVID-19?


title: High Prevalence of Anelloviruses in Vitreous Fluid of Children With Seasonal Hyperacute Panuveitis

text: Seasonal hyperacute panuveitis (SHAPU) is a potentially blinding ocular disease occurring in Nepal that principally affects young children. Random amplification of partially purified vitreous fluid (VF)–derived nucleic acid revealed the presence of human anelloviruses in VF of SHAPU patients. In a comparative study of patients with different ocular pathologies, SHAPU patients were at highest risk of harboring anelloviruses in their eyes. The majority of SHAPU patients had multiple anelloviruses in their VF. The ocular anellovirus load in SHAPU and non-SHAPU patients did not differ and no SHAPU-specific anellovirus variant was detected. Analysis of paired serum and VF samples from SHAPU and non-SHAPU patients showed that the anellovirus detected in VF samples most likely originated from the systemic viral pool during viremia, potentially through breakdown of the blood-ocular barrier. The detection of anelloviruses in VF samples of uveitis patients, profoundly so in SHAPU patients, is imperative and warrants elucidation of its clinical significance.


title: Cellular Proteins in Influenza Virus Particles

text: Virions are thought to contain all the essential proteins that govern virus egress from the host cell and initiation of replication in the target cell. It has been known for some time that influenza virions contain nine viral proteins; however, analyses of other enveloped viruses have revealed that proteins from the host cell can also be detected in virions. To address whether the same is true for influenza virus, we used two complementary mass spectrometry approaches to perform a comprehensive proteomic analysis of purified influenza virus particles. In addition to the aforementioned nine virus-encoded proteins, we detected the presence of 36 host-encoded proteins. These include both cytoplasmic and membrane-bound proteins that can be grouped into several functional categories, such as cytoskeletal proteins, annexins, glycolytic enzymes, and tetraspanins. Interestingly, a significant number of these have also been reported to be present in virions of other virus families. Protease treatment of virions combined with immunoblot analysis was used to verify the presence of the cellular protein and also to determine whether it is located in the core of the influenza virus particle. Immunogold labeling confirmed the presence of membrane-bound host proteins on the influenza virus envelope. The identification of cellular constituents of influenza virions has important implications for understanding the interactions of influenza virus with its host and brings us a step closer to defining the cellular requirements for influenza virus replication. While not all of the host proteins are necessarily incorporated specifically, those that are and are found to have an essential role represent novel targets for antiviral drugs and for attenuation of viruses for vaccine purposes.


docid: yjcjwczl

title: Preparing for a responsible lockdown exit strategy

text: In just a few weeks’ time, leaders across the globe will have to start making decisions about lifting lockdown policies, with considerable social, economic and political consequences. We propose a framework for what is arguably the most difficult health challenge that governments have faced since the beginning of this century: a responsible lockdown exit strategy.


docid: fyqxablr

title: Protecting the gains of malaria elimination in China


title: Aetiology of paediatric pneumonia after the introduction of pneumococcal conjugate vaccine

text: We describe the aetiology of community-acquired pneumonia in children before and after the introduction of the pneumococcal conjugate vaccination (PCV) programme in 2006. Prospective studies were conducted in 2001–2002 (pre-vaccine) and 2009–2011 (post-vaccine) of children aged 0–16 years with radiologically confirmed pneumonia seen in hospital. Investigations included culture, serology, immunofluorescence antibody and urine antigen testing, with an increased use of PCR assays and expanded panels of pathogens in the post-vaccine study. 241 and 160 children were enrolled in the pre- and post-vaccine studies, respectively (73% aged <5 years). Identification of a causative pathogen was higher post-vaccination (61%) than pre-vaccination (48.5%) (p=0.019). Rates of bacterial infections were not different between post- and pre-vaccine studies (17.5% versus 24%, p=0.258). Viral (31%) and mixed (12.5%) infections were found more often post-vaccination (19.5%, p=0.021) than pre-vaccination (5%, p=0.015). Rates of identified pneumococcal infections were comparable between pre- and post-vaccine studies (14.7% versus 17.4%, p=0.557). Diagnosis of pneumococcal infection post-vaccination improved when PCR was used compared to culture (21.6% versus 6%, p=0.0004). Serotypes included in PCV13 but not PCV7 were identified in 75% (18 out of 24) post-vaccination. Infection with nonvaccine pneumococcal serotypes continues to be a significant cause of pneumonia in children in the UK.


title: A Toolkit for Rapid Modular Construction of Biological Circuits in Mammalian Cells

text: The ability to rapidly assemble and prototype cellular circuits is vital for biological research and its applications in biotechnology and medicine. Current methods that permit the assembly of DNA circuits in mammalian cells are laborious, slow, expensive and mostly not permissive of rapid prototyping of constructs. Here we present the Mammalian ToolKit (MTK), a Golden Gate-based cloning toolkit for fast, reproducible and versatile assembly of large DNA vectors and their implementation in mammalian models. The MTK consists of a curated library of characterized, modular parts that can be easily mixed and matched to combinatorially assemble one transcriptional unit with different characteristics, or a hierarchy of transcriptional units weaved into complex circuits. MTK renders many cell engineering operations facile, as showcased by our ability to use the toolkit to generate single-integration landing pads, to create and deliver libraries of protein variants and sgRNAs, and to iterate through Cas9-based prototype circuits. As a biological proof of concept, we used the MTK to successfully design and rapidly construct in mammalian cells a challenging multicistronic circuit encoding the Ebola virus (EBOV) replication complex. This construct provides a non-infectious biosafety level 2 (BSL2) cellular assay for exploring the transcription and replication steps of the EBOV viral life cycle in its host. Its construction also demonstrates how the MTK can enable important and time sensitive applications such as the rapid testing of pharmacological inhibitors of emerging BSL4 viruses that pose a major threat to human health.


The time is now: expedited HIV differentiated service delivery during the COVID‐19 pandemic


title: Clinical Review: Point-of-Care Ultrasound in the Evaluation of COVID-19

text: Abstract The novel coronavirus SARS-CoV-2 has caused a pandemic overwhelming healthcare systems around the world. Hospitals around the world, including the United States, have been struggling to adapt to the influx of patients with COVID-19, the illness caused by SARS-CoV2, given limited resources and high demand for medical care. Point of care ultrasound provides numerous benefits to emergency providers caring for patients with COVID-19 including decreasing resource utilization, assisting in diagnosis, guiding management of the critically ill patient, and aiding in rapid triage of patients under investigations for COVID-19. The studies currently being published have established a typical set of ultrasound findings in COVID-19. Point of care lung ultrasound is rapid and accessible in most emergency departments in the United States and even if many resource-poor settings. This article seeks to provide emergency physicians with a guide to sonographic findings in COVID-19 and an algorithm by which point of care lung ultrasound may assist emergency physicians caring for these patients during the SARS-CoV-2 pandemic.


title: Ramadan Fasting and Risk of Covid-19

text: Almost all religions recommend periods of fasting. Many adult Muslims fast during the holy month of Ramadan each year. Ramadan fasting as a type of intermittent fasting is a non-pharmacological intervention refining the overall health. This year, Ramadan is coincided with coronavirus disease 2019 (COVID-19) outbreak making it one of the most challenging fasting periods for Muslims in the world. There is no solid direct evidence to suggest any adverse effect of Ramadan fasting during the COVID-19 pandemic in healthy individuals. However, there are exemptions in Ramadan Fasting and those at risk of health issues should not fast. COVID-19 is a new disease and there is limited studies concerning its risk factors. The purpose of this review was shedding more light on the potential mechanisms involved in influence of practice of fasting in all forms, including Ramadan fasting on the vulnerability to infection.


title: Coronavirus Disease-2019 and Telehealth: Prevention of Exposure in a Medically Complex Patient With a Mild Presentation

text: The novel coronavirus disease-2019 (COVID-19) was declared a pandemic by the World Health Organization on March 11, 2020. We discuss a less common presentation of COVID-19 in a patient where the use of telemedicine technology prevented the exposure of the healthcare team in an academic general pediatrics clinic. A medically complex 20-year-old male presented via virtual visit for unilateral eye redness and discharge. He received topical ophthalmic antibiotics for presumed acute bacterial conjunctivitis and was counseled on return precautions. Three days later, the patient developed shortness of breath, fever to 102.6°F, and chest pain. COVID-19 testing returned positive. Conjunctivitis is a presenting symptom in 32% of COVID-19 cases. Current evidence suggests that these patients may transmit their illness at the time of presentation. Using telehealth, we avoided exposure to a COVID-19–positive patient who would not have met criteria for testing or droplet precautions. Telehealth allows providers to triage patients, identifying those who need to be seen in person, thereby minimizing exposure. Transitioning toward virtual practice, particularly during a pandemic, prepares for the possibility of healthcare workers being quarantined but still able to provide care and minimizes exposure to asymptomatic patients or those with atypical symptoms.


title: Recommendations on vaccination for Asian small animal practitioners: a report of the WSAVA Vaccination Guidelines Group

text: In 2012 and 2013, the World Small Animal Veterinary Association (WSAVA) Vaccination Guidelines Group (VGG) undertook fact‐finding visits to several Asian countries, with a view to developing advice for small companion animal practitioners in Asia related to the administration of vaccines to dogs and cats. The VGG met with numerous first opinion practitioners, small animal association leaders, academic veterinarians, government regulators and industry representatives and gathered further information from a survey of almost 700 veterinarians in India, China, Japan and Thailand. Although there were substantial differences in the nature and magnitude of the challenges faced by veterinarians in each country, and also differences in the resources available to meet those challenges, overall, the VGG identified insufficient undergraduate and postgraduate training in small companion animal microbiology, immunology and vaccinology. In most of the countries, there has been little academic research into small animal infectious diseases. This, coupled with insufficient laboratory diagnostic support, has limited the growth of knowledge concerning the prevalence and circulating strains of key infectious agents in most of the countries visited. Asian practitioners continue to recognise clinical infections that are now considered uncommon or rare in western countries. In particular, canine rabies virus infection poses a continuing threat to animal and human health in this region. Both nationally manufactured and international dog and cat vaccines are variably available in the Asian countries, but the product ranges are small and dominated by multi‐component vaccines with a licensed duration of immunity (DOI) of only 1 year, or no description of DOI. Asian practitioners are largely unaware of current global trends in small animal vaccinology or of the WSAVA vaccination guidelines. Consequently, most practitioners continue to deliver annual revaccination with both core and non‐core vaccines to adult animals, with little understanding that “herd immunity” is more important than frequent revaccination of individual animals within the population. In this paper, the VGG presents the findings of this project and makes key recommendations for the Asian countries. The VGG recommends that (1) Asian veterinary schools review and increase as needed the amount of instruction in small animal vaccinology within their undergraduate curriculum and increase the availability of pertinent postgraduate education for practitioners; (2) national small animal veterinary associations, industry veterinarians and academic experts work together to improve the scientific evidence base concerning small animal infectious diseases and vaccination in their countries; (3) national small animal veterinary associations take leadership in providing advice to practitioners based on improved local knowledge and global vaccination guidelines; (4) licensing authorities use this enhanced evidence base to inform and support the registration of improved vaccine product ranges for use in their countries, ideally with DOI for core vaccines similar or equal to those of equivalent products available in western countries (i.e. 3 or 4 years). The VGG also endorses the efforts made by Asian governments, non‐governmental organisations and veterinary practitioners in working towards the goal of global elimination of canine rabies virus infection. In this paper, the VGG offers both a current pragmatic and future aspirational approach to small animal vaccination in Asia. As part of this project, the VGG delivered continuing education to over 800 Asian practitioners at seven events in four countries. Accompanying this document is a list of 80 frequently asked questions (with answers) that arose during these discussions. The VGG believes that this information will be of particular value to Asian veterinarians as they move towards implementing global trends in small companion animal vaccinology.


title: Simple Device Design for Plume Management after Pneumoperitoneum in Laparoscopy in COVID-19 Outbreak

text: The impact of COVID-19 on surgical practice is worldwide. Controversy is there regarding dissemination of coronavirus by surgical smoke during laparoscopic surgery; hence, laparoscopic surgeries are being used with great cautions. We propose the use of a simple device, which can be prepared cheaply with material readily available in our hospitals, to manage the surgical plume generated during laparoscopic procedures. With proper management of the surgical plume with the above-proposed device, the concerns regarding aerosol generation can be alleviated and all the benefits of laparoscopy can be extended to our patient in need of a surgery that cannot be delayed.


title: Turnover Intention among Field Epidemiologists in South Korea

text: The purpose of this study was to explore the level of occupational stress, job satisfaction, and turnover intention among Korean field epidemiologists, and to identify the factors that contribute to their turnover intention. We surveyed the Korean field epidemiologists in the cohort from 2016 to 2018 using the Occupational Stress Inventory, revised edition, and questionnaires developed from the Public Health Workforce Interest and Needs Survey. Fisher’s exact test was used to identify the association between sociodemographic characteristics, occupational stress, job satisfaction, and turnover intention. Overall, 17 Korean field epidemiologists participated in this study (response rate: 74%). More than half of field epidemiologists had turnover intention (53%), and it was less likely to be present in the field epidemiologists recruited from the civilian sector than those recruited from the military (adjusted odds ratio, 0.59; 95% confidence interval, 0.39–0.88). Furthermore, about two-thirds of field epidemiologists had a burden of occupational stress on Role Ambiguity (65%), and only one respondent expressed satisfaction with the job. There was no significant relation among the levels of occupational stress, job satisfaction, and turnover intention. In this study, the field epidemiologists recruited from the military were more likely to have turnover intention. Additional studies to identify possible ways to reduce turnover intention among the public health workforce are warranted.


title: Choosing not to follow rules that will reduce the spread of COVID-19

text: In response to the COVID-19 pandemic, many authorities have implemented public health measures that place restrictions on individuals. Understanding how individuals respond to these new rules, particularly whether they are likely to follow or break them, is extremely important. Relational frame theory offers unique insights into rule-governed behavior, allowing researchers to develop functional-analytic interpretations of why a listener may understand a rule, have the required response established in their behavioral repertoire, and still choose not to follow the rule. Drawing from research on rule-following in accordance with relational frame theory and cognitive neuroscience, social psychology, and health literature, this paper presents reasons why a rule may be understood but not followed, identifying important considerations for implementing public health measures to reduce the spread of COVID-19. Specifically, rule-givers should attend to their credibility, authority and ability to mediate consequences, rule plausibility, establishing adequate motivative augmental control, whether the behavior specified in the rule opposes habits, and whether the message incites counterpliance.


title: Use of face coverings by the public during the COVID-19 pandemic: an observational study

text: Public health agencies have recommended that the public wear face coverings as a major non-pharmaceutical intervention to mitigate COVID-19 transmission. However, there is concern whether or not the public has adopted this recommendation. An observational study of 3,271 members of the public was conducted during the COVID-19 pandemic and examined the use of face coverings at grocery stores across 20 Wisconsin counties between May 16th and June 1st, 2020. Of the total individuals observed, we found that only 41.2% used face coverings. Individuals who appeared to be adults (aOR = 1.65; 95% CI = 1.20–2.27) or older adults (aOR = 3.67; 95% CI = 2.59–5.19) were more likely to be wearing face coverings than younger individuals. Additionally, individuals with female gender expression (aOR = 1.43; 95% CI = 1.24–1.66) and individuals shopping at a more expensive grocery store (aOR = 1.95; 95% CI = 1.69–2.25) were more likely to be wearing face coverings. We did not find an association between county level prevalence of COVID-19 cases and face covering use. To our knowledge, this is the first study using direct observations to examine face covering behavior by the public and our findings have implications for public health agencies.


title: Rapid PCR/ESI-MS-based molecular genotyping of Staphylococcus aureus from nasal swabs of emergency department patients

text: BACKGROUND: A limitation of both culture-based and molecular methods of screening for staphylococcal infection is that current tests determine only the presence or absence of colonization with no information on the colonizing strain type. A technique that couples polymerase chain reaction to mass spectrometry (PCR/ESI-MS) has recently been developed and an assay validated to identify and genotype S. aureus and coagulase-negative staphylococci (CoNS). METHODS: This study was conducted to determine the rates, risk factors, and molecular genotypes of colonizing Staphylococcus aureus in adult patients presenting to an inner-city academic emergency department. Participants completed a structured questionnaire to assess hospital and community risks for infection with methicillin-resistant S. aureus (MRSA). Nasal swabs were analyzed by PCR/ESI-MS to identify and genotype S. aureus and CoNS. RESULTS: Of 200 patients evaluated, 59 were colonized with S. aureus; 27 of these were methicillin-resistant strains. Twenty-four of the 59 S. aureus carriers were co-colonized with a CoNS and 140 of the 200 patients were colonized exclusively with CoNS. The molecular genotypes of the 59 S. aureus strains were diverse; 21 unique molecular genotypes belonging to seven major clonal complexes were identified. Eighty-five of 200 patients carried strains with high-level mupirocin resistance. Of these eighty-five participants, 4 were colonized exclusively with S. aureus, 16 were co-colonized with S. aureus and CoNS, and 65 were colonized exclusively with CoNS. CONCLUSION: The prevalence of S. aureus and methicillin-resistant S. aureus colonization in a random sample of patients seeking care in Emergency Department was 29.5% and 13.5%, respectively. A substantial fraction of the S. aureus-colonized patients were co-colonized with CoNS and high-level mupirocin-resistant CoNS. Determining the molecular genotype of S. aureus during intake screening may prove valuable in the future if certain molecular genotypes become associated with increased infection risk.


title: Are pangolins the intermediate host of the 2019 novel coronavirus (2019-nCoV) ?

text: The outbreak of 2019-nCoV pneumonia (COVID-19) in the city of Wuhan, China has resulted in more than 70,000 laboratory confirmed cases, and recent studies showed that 2019-nCoV (SARS-CoV-2) could be of bat origin but involve other potential intermediate hosts. In this study, we assembled the genomes of coronaviruses identified in sick pangolins. The molecular and phylogenetic analyses showed that pangolin Coronaviruses (pangolin-CoV) are genetically related to both the 2019-nCoV and bat Coronaviruses but do not support the 2019-nCoV arose directly from the pangolin-CoV. Our study also suggested that pangolin be natural host of Betacoronavirus, with a potential to infect humans. Large surveillance of coronaviruses in pangolins could improve our understanding of the spectrum of coronaviruses in pangolins. Conservation of wildlife and limits of the exposures of humans to wildlife will be important to minimize the spillover risks of coronaviruses from wild animals to humans.


title: Higher tourism specialization, better hotel industry efficiency?

text: This study examines the effects of both specialization in tourism and market competition on the efficiency of the hotel industry. For this purpose, stochastic frontier analysis (SFA) was employed to evaluate the efficiency of the hotel industry on a provincial level in China and to analyze how it is impacted by specialization in tourism and market competition. The results confirm that tourism specialization and market competition exert a synergistic effect on hotel industry efficiency in China. This study finds that tourism development as represented by a high level of tourism specialization by a destination does not guarantee high efficiency in hotels but does enhance the negative effect of market competition on hotel industry efficiency. Significant policy and managerial implications stem from these findings.


title: First-wave COVID-19 transmissibility and severity in China outside Hubei after control measures, and second-wave scenario planning: a modelling impact assessment

text: BACKGROUND: As of March 18, 2020, 13 415 confirmed cases and 120 deaths related to coronavirus disease 2019 (COVID-19) in mainland China, outside Hubei province—the epicentre of the outbreak—had been reported. Since late January, massive public health interventions have been implemented nationwide to contain the outbreak. We provide an impact assessment of the transmissibility and severity of COVID-19 during the first wave in mainland Chinese locations outside Hubei. METHODS: We estimated the instantaneous reproduction number (R(t)) of COVID-19 in Beijing, Shanghai, Shenzhen, Wenzhou, and the ten Chinese provinces that had the highest number of confirmed COVID-19 cases; and the confirmed case-fatality risk (cCFR) in Beijing, Shanghai, Shenzhen, and Wenzhou, and all 31 Chinese provinces. We used a susceptible–infectious–recovered model to show the potential effects of relaxing containment measures after the first wave of infection, in anticipation of a possible second wave. FINDINGS: In all selected cities and provinces, the R(t) decreased substantially since Jan 23, when control measures were implemented, and have since remained below 1. The cCFR outside Hubei was 0·98% (95% CI 0·82–1·16), which was almost five times lower than that in Hubei (5·91%, 5·73–6·09). Relaxing the interventions (resulting in R(t) >1) when the epidemic size was still small would increase the cumulative case count exponentially as a function of relaxation duration, even if aggressive interventions could subsequently push disease prevalence back to the baseline level. INTERPRETATION: The first wave of COVID-19 outside of Hubei has abated because of aggressive non-pharmaceutical interventions. However, given the substantial risk of viral reintroduction, particularly from overseas importation, close monitoring of R(t) and cCFR is needed to inform strategies against a potential second wave to achieve an optimal balance between health and economic protection. FUNDING: Health and Medical Research Fund, Hong Kong, China.


title: Dynamics: the missing link between structure and function of the viral RNA-dependent RNA polymerase?

text: The structural basis for nucleotide incorporation fidelity remains an open question for all nucleic acid polymerases. Addressing this question for the viral RNA-dependent RNA polymerase (RdRp) is of particular, practical significance because it is a determinant of sensitivity to antiviral nucleosides and may be a determinant of viral virulence. All polymerases are thought to employ the same catalytic mechanism, but the rate of nucleotide incorporation can vary substantially. Here we review some of the recent work with the RdRp that leads us to suggest that structure provides only a partial understanding of RdRp function and dynamics may be the missing link.


title: Innovation Lessons From the COVID-19 Pandemic

text: Abstract On December 31, 2019, Chinese authorities reported the first case of what would become known as COVID-19 to the World Health Organization (WHO), which declared a Public Health Emergency of International Concern one month later. In the ensuing weeks, health care organizations, industry partners, and government agencies worldwide have quickly come together to address pandemic challenges such as scaling high-quality laboratory tests, conducting treatment research, and enabling virtual health care in a time of social distancing. What we have accomplished together to date demonstrates the opportunity for future cross-sector partnerships in science and technology to fight the disease, while simultaneously creating the new digital health care solutions that our world greatly needs.


Editorial: Postcards from War Zones—Personal Experiences During the COVID-19 Pandemic


title: Impact of the Perceived Threat of COVID-19 on Variety-seeking

text: Abstract The COVID-19 pandemic has significantly influenced our daily and social lives as well as our consumption patterns. This paper focuses on the relationship between the COVID-19 threat and variety-seeking. Based on several theories, including reactance theory and terror management theory, we predict that the perceived threat of COVID-19 will increase the tendency to choose more and different options in multiple choice settings. Firstly, two empirical studies demonstrate that variety-seeking in food and stationery choices is enhanced as people's perceived threat from the disease increases. Study 3 further suggests the boundary conditions of the above pattern in that the type of decision (i.e., multiple option selections across different brands vs. within the same brand) will moderate the impact of the perceived threat on varietyseeking. Specifically, when the decision involved choice across different brands, participants showed higher variety-seeking under high (vs. low) perceived threat. However, the opposite pattern was true when the decision involved choice within the same brand. This research offers a deeper understanding of how variety-seeking can be changed by the perceived threat of COVID-19.


title: Molecular and serological investigation of 2019-nCoV infected patients: implication of multiple shedding routes

text: In December 2019, a novel coronavirus (2019-nCoV) caused an outbreak in Wuhan, China, and soon spread to other parts of the world. It was believed that 2019-nCoV was transmitted through respiratory tract and then induced pneumonia, thus molecular diagnosis based on oral swabs was used for confirmation of this disease. Likewise, patient will be released upon two times of negative detection from oral swabs. However, many coronaviruses can also be transmitted through oral–fecal route by infecting intestines. Whether 2019-nCoV infected patients also carry virus in other organs like intestine need to be tested. We conducted investigation on patients in a local hospital who were infected with this virus. We found the presence of 2019-nCoV in anal swabs and blood as well, and more anal swab positives than oral swab positives in a later stage of infection, suggesting shedding and thereby transmitted through oral–fecal route. We also showed serology test can improve detection positive rate thus should be used in future epidemiology. Our report provides a cautionary warning that 2019-nCoV may be shed through multiple routes.


title: Proteomic alteration of PK-15 cells after infection by porcine circovirus type 2

text: Porcine circovirus type 2 (PCV2) has been identified as the essential causal agent of post-weaning multisystemic wasting syndrome, which has spread worldwide. To discover cellular protein responses of PK-15 cells to PCV2 infection, two-dimensional liquid chromatography–tandem mass spectrometry (MS) coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling was employed to quantitatively identify the proteins that were differentially expressed in PK-15 from the PCV2-infected group compared to the uninfected control group. A total of 196 cellular proteins in PK-15 that were significantly altered at different time periods post-infection were identified. These differentially expressed proteins were related to the biological processes of binding, cell structure, signal transduction, cell adhesion, etc. and their interactions. Moreover, some of these proteins were further confirmed by Western blot. The high number of differentially expressed proteins identified should be very useful in elucidating the mechanism of replication and pathogenesis of PCV2 in the future.


title: Environmental effects

text: Our pandemic response is cutting emissions, but it isn't a climate change fix


title: Effectiveness of handwashing in preventing SARS: a review

text: This review examines the literature, including literature in Chinese, on the effectiveness of handwashing as an intervention against severe acute respiratory syndrome (SARS) transmission. Nine of 10 epidemiological studies reviewed showed that handwashing was protective against SARS when comparing infected cases and non‐infected controls in univariate analysis, but only in three studies was this result statistically significant in multivariate analysis. There is reason to believe that this is because most of the studies were too small. The evidence for the effectiveness of handwashing as a measure against SARS transmission in health care and community settings is suggestive, but not conclusive.


Private Equity-Backed Hospital Investments and the Impact of the COVID-19 epidemic


title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

text: Abstract Middle East respiratory syndrome (MERS) represents an important respiratory disease accompanied by lethal outcome in one third of human patients. In recent years, several investigators developed protective antibodies which could be used as prophylaxis in prospective human epidemics. In the current study, eight human monoclonal antibodies (mAbs) with neutralizing and non-neutralizing capabilities, directed against different epitopes of the MERS-coronavirus (MERS-CoV) spike (MERS-S) protein, were investigated with regard to their ability to immunohistochemically detect respective epitopes on formalin-fixed paraffin-embedded (FFPE) nasal tissue sections of MERS-CoV experimentally infected alpacas. The most intense immunoreaction was detected using a neutralizing antibody directed against the receptor binding domain S1B of the MERS-S protein, which produced an immunosignal in the cytoplasm of ciliated respiratory epithelium and along the apical membranous region. A similar staining was obtained by two other mAbs which recognize the sialic acid-binding domain and the ectodomain of the membrane fusion subunit S2, respectively. Five mAbs lacked immunoreactivity for MERS-CoV antigen on FFPE tissue, even though they belong, at least in part, to the same epitope group. In summary, three tested human mAbs demonstrated capacity for detection of MERS-CoV antigen on FFPE samples and may be implemented in double or triple immunohistochemical methods.


docid: lmzkseiz

title: Effects of non-pharmaceutical interventions on COVID-19 cases, deaths, and demand for hospital services in the UK: a modelling study

text: BACKGROUND: Non-pharmaceutical interventions have been implemented to reduce transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the UK. Projecting the size of an unmitigated epidemic and the potential effect of different control measures has been crucial to support evidence-based policy making during the early stages of the epidemic. This study assesses the potential impact of different control measures for mitigating the burden of COVID-19 in the UK. METHODS: We used a stochastic age-structured transmission model to explore a range of intervention scenarios, tracking 66·4 million people aggregated to 186 county-level administrative units in England, Wales, Scotland, and Northern Ireland. The four base interventions modelled were school closures, physical distancing, shielding of people aged 70 years or older, and self-isolation of symptomatic cases. We also modelled the combination of these interventions, as well as a programme of intensive interventions with phased lockdown-type restrictions that substantially limited contacts outside of the home for repeated periods. We simulated different triggers for the introduction of interventions, and estimated the impact of varying adherence to interventions across counties. For each scenario, we projected estimated new cases over time, patients requiring inpatient and critical care (ie, admission to the intensive care units [ICU]) treatment, and deaths, and compared the effect of each intervention on the basic reproduction number, R(0). FINDINGS: We projected a median unmitigated burden of 23 million (95% prediction interval 13–30) clinical cases and 350 000 deaths (170 000–480 000) due to COVID-19 in the UK by December, 2021. We found that the four base interventions were each likely to decrease R(0), but not sufficiently to prevent ICU demand from exceeding health service capacity. The combined intervention was more effective at reducing R(0), but only lockdown periods were sufficient to bring R(0) near or below 1; the most stringent lockdown scenario resulted in a projected 120 000 cases (46 000–700 000) and 50 000 deaths (9300–160 000). Intensive interventions with lockdown periods would need to be in place for a large proportion of the coming year to prevent health-care demand exceeding availability. INTERPRETATION: The characteristics of SARS-CoV-2 mean that extreme measures are probably required to bring the epidemic under control and to prevent very large numbers of deaths and an excess of demand on hospital beds, especially those in ICUs. FUNDING: Medical Research Council.


title: Development of a RT Real-Time PCR for the Detection and Quantification of Human Rhinoviruses

text: Human Rhinoviruses (HRV) are the most common viral agents, being responsible for upper as well as lower respiratory tract infections. Evidence demonstrating that HRV disease is not exclusively limited to the upper airways and may cause lower respiratory complications, together with the frequency of HRV infections and the increasing number of immunocompromised patients underline the need for including HRV in virological diagnostics of acute lower respiratory tract illness. This article describes the development and optimization of a reverse transcription (RT) real-time PCR assay for quantification of HRV RNA in clinical samples. Efficiency, sensitivity, specificity, inter- and intra-assay variability, and dynamic range have been determined. Subsequently, the assay has been validated on bronchoalveolar lavage (BAL) specimens obtained from immunocompetent and immunocompromised patients.


The camel, new challenges for a sustainable development


title: Acute kidney injury in pregnant women following SARS-CoV-2 infection: A case report from Iran

text: We reported a 33-year-old female case with novel coronavirus disease 2019 (COVID-19) accompanied by Acute tubular necrosis (ATN). She had a gestational age of 34 weeks. The patient referred to treatment clinic for COVID-19 in Imam Reza hospital of Tabriz (Iran) after having flu-like symptoms. In radiologic assessment, ground glass opacity (GGO) with consolidation was found in upper right lobe. Lopinavir/ritonavir (200mg/50mg) two tablet tow times, Ribavirin 200mg every six hours, and Oseltamivir 75mg tow times were given for the treatment of COVID-19. The medications used for treatment of pneumonia were Meropenem, Ciprofloxacin, Vancomycin. All doses of medications were administrated by adjusted dose assuming the patient is anephric. Also, a few supplements were also given after ATN development including daily Rocaltrol and Nephrovit (as a multivitamin appropriate for patients with renal failure), Folic acid and Calcium carbonate. The patient is still under ventilator with a Fraction of inspired oxygen (FiO(2)) of 60% and Positive end-expiratory pressure (PEEP) of eight. SpO(2) is 94% but the patient's ATN problem has been resolved. We started weaning from mechanical ventilator. The patient is conscious with full awareness to time, person and place. The maternal well-being is achieved and her neonate was discharged.


title: To clear or to fear: An innate perspective on factor VIII immunity

text: Abstract The enigma that is factor VIII immunogenicity remains ever pertinent in the treatment of hemophilia A. Development of neutralizing antibodies against the therapeutic protein in 25–30% of patients likely depends on the appropriate activation of the innate immune response shortly following antigen encounter. Our understanding of this important immunological synapse remains ill-defined. In this review, we examine the three distinct factors contributing to the fate of factor VIII almost immediately after infusion: the characteristics of the protein, the cell, and the microenvironment. We propose a continuum between clearance and antigen presentation that facilitates removal of FVIII from circulation leading to either tolerance or immunity.


title: Body Recovery

text: Abstract Appropriate precautions are critical to minimize damage to the body during recovery and removal from a scene and transit to the mortuary to allow for accurate interpretation of injury and the minimization of contamination. Many elements of the investigation may be affected by the way in which tasks are conducted. Members of numerous jurisdictions must all work together to ensure the best outcome both from a forensic point of view and also in preserving and respecting the dignity of the deceased. This chapter will outline the techniques necessary for preserving the body and associated evidence during transport, including specific considerations for different types of cases.


title: Securing Posterior Auricular Incision with Button Headbands in Prominent Ear Patients Wearing Surgical Masks for Self-protection in the Pandemic

text: ABSTRACT: In this letter, we describe an attachment on conventional headbands recommended for the postoperative period in prominent ear patients to confidently wear surgical masks without disturbing the posterior auricular incision. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.


title: Molecular Epidemiology of Hospital Outbreak of Middle East Respiratory Syndrome, Riyadh, Saudi Arabia, 2014

text: We investigated an outbreak of Middle East respiratory syndrome (MERS) at King Fahad Medical City (KFMC), Riyadh, Saudi Arabia, during March 29–May 21, 2014. This outbreak involved 45 patients: 8 infected outside KFMC, 13 long-term patients at KFMC, 23 health care workers, and 1 who had an indeterminate source of infection. Sequences of full-length MERS coronavirus (MERS-CoV) from 10 patients and a partial sequence of MERS-CoV from another patient, when compared with other MERS-CoV sequences, demonstrated that this outbreak was part of a larger outbreak that affected multiple health care facilities in Riyadh and possibly arose from a single zoonotic transmission event that occurred in December 2013 (95% highest posterior density interval November 8, 2013–February 10, 2014). This finding suggested continued health care–associated transmission for 5 months. Molecular epidemiology documented multiple external introductions in a seemingly contiguous outbreak and helped support or refute transmission pathways suspected through epidemiologic investigation.


title: Preventing Airborne Disease Transmission: Review of Methods for Ventilation Design in Health Care Facilities

text: Health care facility ventilation design greatly affects disease transmission by aerosols. The desire to control infection in hospitals and at the same time to reduce their carbon footprint motivates the use of unconventional solutions for building design and associated control measures. This paper considers indoor sources and types of infectious aerosols, and pathogen viability and infectivity behaviors in response to environmental conditions. Aerosol dispersion, heat and mass transfer, deposition in the respiratory tract, and infection mechanisms are discussed, with an emphasis on experimental and modeling approaches. Key building design parameters are described that include types of ventilation systems (mixing, displacement, natural and hybrid), air exchange rate, temperature and relative humidity, air flow distribution structure, occupancy, engineered disinfection of air (filtration and UV radiation), and architectural programming (source and activity management) for health care facilities. The paper describes major findings and suggests future research needs in methods for ventilation design of health care facilities to prevent airborne infection risk.


COVID-19 Pandemic: International Variation of Personal Protective Equipment and Infection Prevention and Control Guidelines


title: Exploring the binding mechanism of the main proteinase in SARS-associated coronavirus and its implication to anti-SARS drug design

text: Abstract The main proteinase of SARS-associated coronavirus (SARS-CoV) plays an important role in viral transcription and replication, and is an attractive target for anti-SARS drug development. The important thing is to understand its binding mechanism with possible ligands. In this study, we investigated possible noncanonical interactions, potential inhibitors, and binding pockets in the main proteinase of SARS-CoV based on its recently determined crystal structure. These findings provide a wide clue to searching for anti-SARS drug. Interestingly, we found that similar structure patterns exist in SARS-CoV main proteinase with Poliovirus 3c Proteinase, Rhinovirus 3c Protease, Nsp4 Proteinase From Equine Arteritis Virus, Hepatitis C Virus Ns3 Protease, Hepatitis A Virus 3c Protease, and Dengue Virus Ns3 Protease. It suggests that the available drugs in these viruses could be used to fight SARS disease.


title: Können Haustiere Corona auf ihre Besitzer übertragen?: SARS-CoV-2 bei Hunden und Katzen

text: Haustiere können sich mit SARS-CoV-2 infizieren, ohne symptomatisch zu erkranken. Die Sorge, Katzen oder Hunde könnten als „stille Überträger“ Einfluss auf das Pandemiegeschehen nehmen, ist aber zum jetzigen Stand wohl unbegründet. Die Leiterin der Medizinischen Kleintierklinik der LMU München, Prof. Katrin Hartmann, erklärt, warum.


title: A Novel Method for the Capture-based Purification of Whole Viral Native RNA Genomes

text: Current technologies for targeted characterization and manipulation of viral RNA either involve amplification or ultracentrifugation with isopycnic gradients of viral particles to decrease host RNA background. The former strategy is non-compatible for characterizing properties innate to RNA strands such as secondary structure, RNA-RNA interactions, and also for nanopore direct RNA sequencing involving the sequencing of native RNA strands. The latter strategy, ultracentrifugation, causes loss in genomic information due to its inability to retrieve unassembled viral RNA. We developed a novel nucleic acid manipulation technique involving the capture of whole viral native RNA genomes for downstream RNA assays using hybridization baits in solution to circumvent these problems. This technique involves hybridization of biotinylated baits at 500 nucleotides (nt) intervals, stringent washes and release of free native RNA strands using DNase I treatment, with a turnaround time of about 6 h 15 min. Proof of concept was primarily done using RT-qPCR with dengue virus infected Huh-7 cells. We report that this protocol was able to purify viral RNA (561-791 fold). We also describe a successful application of our capture-based purification method to direct RNA sequencing, with a 77.47% of reads mapping to the target viral genome. We observed a reduction in human host RNA background by 1580 fold, a 99.91% recovery of viral genome with at least 15x coverage, and a mean coverage across the genome of 120x. This report is, to the best of our knowledge, the first description of a capture-based purification method for whole viral RNA genomes. The fundamental advantages of using our capture-based purification method makes it a superior alternative to conventional viral purification methods and would potentially pave a new path for the direct characterization and sequencing of native RNA molecules.


title: Porcine Reproductive and Respiratory Syndrome Virus Infection Induces Stress Granule Formation Depending on Protein Kinase R-like Endoplasmic Reticulum Kinase (PERK) in MARC-145 Cells

text: Stress granules (SGs) are sites of mRNA storage that are formed in response to various conditions of stress, including viral infections. Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. In this study, we found that infection of PRRSV strain WUH3 (genotype 2 PRRSV) induced stable formation of robust SGs in MARC-145 cells, as demonstrated by the recruitment of marker proteins of SGs, including TIA1, G3BP1, and eIF3η. Treatment with specific inhibitors or siRNAs against the stress kinases that are involved in SG formation revealed that PRRSV induced SG formation through a PERK (protein kinase R–like endoplasmic reticulum kinase)-dependent mechanism. Impairment of SG assembly by concomitant knockdown of the SG marker proteins (TIA1, G3BP1, and TIAR) did not affect PRRSV growth, while significantly enhanced PRRSV-induced NF-κB subunit p65 phosphorylation and inflammatory cytokine production. Taken together, our results demonstrate that PRRSV induces SG formation via a PERK-dependent pathway and that SGs are involved in the signaling pathway of the PRRSV-induced inflammatory response in MARC-145 cells.


xTAG™ RVP assay: analytical and clinical performance


title: Evaluation of infliximab as second-line treatment of acute graft versus host disease -validating response on day 7 and 28 as predictors of survival

text: Several immunosuppressive drugs have been proposed for second-line treatment of steroid-refractory acute graft versus host disease (aGvHD) after allogeneic hematopoietic stem cell transplantation. However, the studies on these drugs are small, retrospective, uncontrolled and use different endpoints. Therefore, it remains unknown which treatment is superior. We retrospectively evaluated 68 consecutive patients treated with infliximab for aGvHD. We adhered to recently proposed guidelines for aGvHD trials and thus evaluated response on day 7 and 28. Furthermore, we assessed the composite endpoint 6 months freedom from treatment failure (6MFTF). The majority of patients had grade III-IV aGvHD. We found that 41 patients (60%) responded on day 7 and 31 patients (46%) on day 28. Twenty-four patients (35%) achieved 6MFTF. The main reasons for failure within 6 months were death (n = 31) or additional immunosuppression (n = 16). By six and 24 months, 44 and 34% of the patients were alive respectively. Patients with response to infliximab on day 7 and 28 had significantly higher overall survival (OS) probability than non-responders. We show that response on day 7 and 28 identifies high and low risk groups. Patients who fail to respond should be identified early and offered alternative therapy.


title: Cardiopulmonary Resuscitation in the Prone Position: A Good Option for Patients With COVID-19

text: Supplemental Digital Content is available in the text.


title: Single Fluorescence Channel-based Multiplex Detection of Avian Influenza Virus by Quantitative PCR with Intercalating Dye

text: Since its invention in 1985 the polymerase chain reaction (PCR) has become a well-established method for amplification and detection of segments of double-stranded DNA. Incorporation of fluorogenic probe or DNA intercalating dyes (such as SYBR Green) into the PCR mixture allowed real-time reaction monitoring and extraction of quantitative information (qPCR). Probes with different excitation spectra enable multiplex qPCR of several DNA segments using multi-channel optical detection systems. Here we show multiplex qPCR using an economical EvaGreen-based system with single optical channel detection. Previously reported non quantitative multiplex real-time PCR techniques based on intercalating dyes were conducted once the PCR is completed by performing melting curve analysis (MCA). The technique presented in this paper is both qualitative and quantitative as it provides information about the presence of multiple DNA strands as well as the number of starting copies in the tested sample. Besides important internal control, multiplex qPCR also allows detecting concentrations of more than one DNA strand within the same sample. Detection of the avian influenza virus H7N9 by PCR is a well established method. Multiplex qPCR greatly enhances its specificity as it is capable of distinguishing both haemagglutinin (HA) and neuraminidase (NA) genes as well as their ratio.


title: Angiotensin-Converting Enzyme 2: The First Decade

text: The renin-angiotensin system (RAS) is a critical regulator of hypertension, primarily through the actions of the vasoactive peptide Ang II, which is generated by the action of angiotensin-converting enzyme (ACE) mediating an increase in blood pressure. The discovery of ACE2, which primarily metabolises Ang II into the vasodilatory Ang-(1-7), has added a new dimension to the traditional RAS. As a result there has been huge interest in ACE2 over the past decade as a potential therapeutic for lowering blood pressure, especially elevation resulting from excess Ang II. Studies focusing on ACE2 have helped to reveal other actions of Ang-(1-7), outside vasodilation, such as antifibrotic and antiproliferative effects. Moreover, investigations focusing on ACE2 have revealed a variety of roles not just catalytic but also as a viral receptor and amino acid transporter. This paper focuses on what is known about ACE2 and its biological roles, paying particular attention to the regulation of ACE2 expression. In light of the entrance of human recombinant ACE2 into clinical trials, we discuss the potential use of ACE2 as a therapeutic and highlight some pertinent questions that still remain unanswered about ACE2.


title: Rotavirus C: prevalence in suckling piglets and development of virus-like particles to assess the influence of maternal immunity on the disease development

text: Rotavirus C (RVC) has been detected increasingly in humans and swine in different countries, including the US. It is associated with significant economic losses due to diarrheal disease in nursing piglets. In this study we aimed: (1) to determine the prevalence of RVC in healthy and diarrheic suckling piglets on US farms; and (2) to evaluate if maternal antibody (Ab) levels were associated with protection of newborn suckling piglets against RVC. There was a significantly higher prevalence (p = 0.0002) of litters with diarrhea born to gilts compared with those born to multiparous sows. Of 113 nursing piglet fecal samples tested, 76.1% were RVC RNA positive. Fecal RVC RNA was detected in significantly (p = 0.0419) higher quantities and more frequently in piglets with diarrhea compared with healthy ones (82.5 vs. 69.9%). With the exception of the historic strain Cowden (G1 genotype), field RVC strains do not replicate in cell culture, which is a major impediment for studying RVC pathogenesis and immunity. To circumvent this, we generated RVC virus-like particles (VLPs) for Cowden (G1), RV0104 (G3) and RV0143 (G6) and used them as antigens in ELISA to detect swine RVC Abs in serum and milk from the sows. Using RVC-VLP Ab ELISA we demonstrated that sows with diarrheic litters had significantly lower RVC IgA and IgG Ab titers in milk compared to those with healthy litters. Thus, our data suggest that insufficient lactogenic protection provided by gilts plays a key role in the development of and the increased prevalence of clinical RVC disease.