title: Steroid-associated osteonecrosis animal model in rats

text: OBJECTIVE: Established preclinical disease models are essential for not only studying aetiology and/or pathophysiology of the relevant diseases but more importantly also for testing prevention and/or treatment concept(s). The present study proposed and established a detailed induction and assessment protocol for a unique and cost-effective preclinical steroid-associated osteonecrosis (SAON) in rats with pulsed injections of lipopolysaccharide (LPS) and methylprednisolone (MPS). METHODS: Sixteen 24-week-old male Sprague–Dawley rats were used to induce SAON by one intravenous injection of LPS (0.2 mg/kg) and three intraperitoneal injections of MPS (100 mg/kg) with a time interval of 24 hour, and then, MPS (40 mg/kg) was intraperitoneally injected three times a week from week 2 until sacrifice. Additional 12 rats were used as normal controls. Two and six weeks after induction, animals were scanned by metabolic dual energy X-ray absorptiometry for evaluation of tissue composition; serum was collected for bone turnover markers, Microfil perfusion was performed for angiography, the liver was collected for histopathology and bilateral femora and bilateral tibiae were collected for histological examination. RESULTS: Three rats died after LPS injection, i.e., with 15.8% (3/19) mortality. Histological evaluation showed 100% incidence of SAON at week 2. Dual energy X-ray absorptiometry showed significantly higher fat percent and lower lean mass in SAON group at week 6. Micro-computed tomography (Micro-CT) showed significant bone degradation at proximal tibia 6 weeks after SAON induction. Angiography illustrated significantly less blood vessels in the proximal tibia and significantly more leakage particles in the distal tibia 2 weeks after SAON induction. Serum amino-terminal propeptide of type I collagen and osteocalcin were significantly lower at both 2 and 6 weeks after SAON induction, and serum carboxy-terminal telopeptide was significantly lower at 6 weeks after SAON induction. Histomorphometry revealed significantly lower osteoblast surface and higher marrow fat fraction and oedema area in SAON group. Hepatic oedema appeared 2 weeks after SAON induction, and lipid accumulation appeared in the liver of SAON rats 6 weeks after SAON induction. CONCLUSION: The present study successfully induced SAON in rats with pulsed injection of LPS and MPS, which was well simulating the clinical feature and pathology. Apart from available large animal models, such as bipedal emus or quadrupedal rabbits, our current SAON small model in rats could be a cost-effective preclinical experimental model to study body metabolism, molecular mechanism of SAON and potential drugs developed for prevention or treatment of SAON. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The present study successfully induced SAON in a small animal model in rats with pulsed injection of LPS and MPS. The evaluation protocols with typical histopathologic ON features and advanced evaluation approaches to identify the metabolic disorders of SAON could be used in future rat SAON studies. The SAON rat model is a suitable and cost-effective animal model to study molecular mechanism of SAON and potential drugs developed for prevention and treatment of SAON.


A Special Issue on Respiration and the Airway: critical topics at a challenging time


title: Potential negative effects of the free use of chloroquine to manage COVID‐19 in Colombia

text: Coronavirus disease 2019 (COVID-19) pandemic has challenged healthcare systems around the world. Unfortunately, failure has ensued: high-income countries have succumbed to the global emergency despite highly prepared human and technological assets. There is no current consensus on pharmacological management of COVID-19, but chloroquine phosphate (CQ) has emerged as a possible therapeutical candidate. However, no conclusive evidence has been published on the efficacy of CQ against SARS-CoV-2 infection. The Colombian government has approved the use of CQ for COVID-19, a measure that could negatively impact a large number of patients who depend on this drug for the treatment of other life-threatening diseases such as malaria and rheumatic autoimmune diseases. Careful consideration should be taken by Colombian authorities regarding the use of CQ in context of the ongoing pandemic. This article is protected by copyright. All rights reserved.


title: Successful implementation of preventive measures leads to low relevance of SARS‐CoV‐2 in liver transplant patients: Observations from a German outpatient department

text: BACKGROUND: Immunosuppressed liver transplant (LT) patients are considered to be at high risk for any kind of infection. What the outbreak of the novel severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) means for the transplant cohort is a question that, as of now, cannot easily be answered. Data on prevalence, relevance of the novel virus and clinical course of the infection in stable LT patients are limited. METHODS: Nasopharyngeal swabs were performed in our outpatient department during the shutdown between March and April 2020 in Germany. RESULTS: The prevalence of SARS‐CoV‐2 was 3%. Three out of a cohort of 101 LT‐patients were asymptomatic for respiratory diseases. Respiratory complaints were common and not associated with SARS‐CoV‐2 infection. The overall monthly mortality rate was 0,22% and did not show alterations during the shutdown in Germany. CONCLUSIONS: If preventive measures are applied, LT‐patients do not seem to be at a higher risk for SARS‐CoV‐2 infection. Telemedicine in the outpatient setting may help to maintain distance and to reduce direct patient contact. However, standard of care must be guaranteed for patients with relevant comorbidities in spite of pandemics, because complications may arise from preexisting conditions.


title: 3.33 Main-Group Medicinal Chemistry Including Li and Bi*

text: Abstract Main-group element compounds were among the first developed in the modern era as pharmaceutical preparations for the treatment of a wide variety of human ailments; it is now recognized that many of these elements exist in traditional medicine of many societies, for example, arsenic. The use of main-group element compounds in contemporary medicine continues for the treatment of, for example, depression (Li), stomach ulcers (Bi), cancer (As and Ga), and leishmaniasis (Sb). Not surprisingly, new compounds of these elements, and other main-group elements, continue to be investigated for their potential use in new therapies. In this chapter, the use of main-group elements as therapeutic agents is outlined and also, where understood, comments on biological targets and mechanisms of action. Further, key advances in new potential applications of main-group element compounds in medicine are evaluated.


title: Have microbes, will travel Ann Marie Kimball Risky Trade

text: Unknown


title: Small interfering RNA inhibits SARS-CoV nucleocapsid gene expression in cultured cells and mouse muscles

text: SARS-CoV is a newly identified coronavirus that causes severe acute respiratory syndrome (SARS). Currently, there is no effective method available for prophylaxis and treatment of SARS-CoV infections. In the present study, the influence of small interfering RNA (siRNA) on SARS-CoV nucleocapsid (N) protein expression was detected in cultured cells and mouse muscles. Four siRNA expression cassettes driven by mouse U6 promoter targeting SARS-CoV N gene were prepared, and their inhibitory effects on expression of N and enhanced green fluorescence protein (EGFP) fusion protein were observed. A candidate siRNA was proved to down-regulate N and EGFP expression actively in a sequence-specific manner. The expression vector of this siRNA was constructed and confirmed to reduce N and EGFP expression efficiently in both cultured cells and adult mouse muscles. Our findings suggest that the siRNA should provide the basis for prophylaxis and therapy of SARS-CoV infection in human.


title: The triad “dogs, conservation and zoonotic diseases” – An old and still neglected problem in Brazil

text: Abstract The presence of domestic/free-ranging dogs in Brazilian protected areas and native vegetation fragments is an important problem, mainly because these animals pose a threat to wild species that live in such areas. In addition, dogs constantly circulate between wildlife environments and urban regions, acting as “bridges” in spillover events. Dogs are traditionally recognized as vectors of zoonoses, which are correct, but their roles as facilitating agents for the “jump” of pathogens from wild animals to humans (and vice versa) are sparsely debated. In this context, this work briefly describes the different roles of dogs in the dynamics and ecology of infectious diseases, using the Brazilian scenario as a study model.


title: Interferon Lambda Genetics and Biology in Regulation of Viral Control

text: Type III interferons, also known as interferon lambdas (IFNλs), are the most recent addition to the IFN family following their discovery in 2003. Initially, IFNλ was demonstrated to induce expression of interferon-stimulated genes and exert antiviral properties in a similar manner to type I IFNs. However, while IFNλ has been described to have largely overlapping expression and function with type I IFNs, it has become increasingly clear that type III IFNs also have distinct functions from type I IFNs. In contrast to type I IFNs, whose receptor is ubiquitously expressed, type III IFNs signal and function largely at barrier epithelial surfaces, such as the respiratory and gastrointestinal tracts, as well as the blood–brain barrier. In further support of unique functions for type III IFNs, single nucleotide polymorphisms in IFNL genes in humans are strongly associated with outcomes to viral infection. These biological linkages have also been more directly supported by studies in mice highlighting roles of IFNλ in promoting antiviral immune responses. In this review, we discuss the current understanding of type III IFNs, and how their functions are similar to, and different from, type I IFN in various immune cell subtypes and viral infections.


title: Further evidence that chilblains are a cutaneous manifestation of COVID‐19 infection

text: A range of cutaneous conditions including chilblain‐like lesions have been reported in patients with Coronavirus 2019 (COVID‐19) due to severe acute respiratory Syndrome coronavirus 2 (SARS‐Cov‐2). Using clinical data and images, the authors of a recent nationwide Spanish survey identified 5 clinical patterns which were associated with different patient demographics, onset (timing), and prognosis.


title: An IDEA for Short Term Outbreak Projection: Nearcasting Using the Basic Reproduction Number

text: BACKGROUND: Communicable disease outbreaks of novel or existing pathogens threaten human health around the globe. It would be desirable to rapidly characterize such outbreaks and develop accurate projections of their duration and cumulative size even when limited preliminary data are available. Here we develop a mathematical model to aid public health authorities in tracking the expansion and contraction of outbreaks with explicit representation of factors (other than population immunity) that may slow epidemic growth. METHODOLOGY: The Incidence Decay and Exponential Adjustment (IDEA) model is a parsimonious function that uses the basic reproduction number R(0), along with a discounting factor to project the growth of outbreaks using only basic epidemiological information (e.g., daily incidence counts). PRINCIPAL FINDINGS: Compared to simulated data, IDEA provides highly accurate estimates of total size and duration for a given outbreak when R(0) is low or moderate, and also identifies turning points or new waves. When tested with an outbreak of pandemic influenza A (H1N1), the model generates estimated incidence at the i+1(th) serial interval using data from the i(th) serial interval within an average of 20% of actual incidence. CONCLUSIONS AND SIGNIFICANCE: This model for communicable disease outbreaks provides rapid assessments of outbreak growth and public health interventions. Further evaluation in the context of real-world outbreaks will establish the utility of IDEA as a tool for front-line epidemiologists.


title: A study on antigenicity and receptor-binding ability of fragment 450–650 of the spike protein of SARS coronavirus

text: Abstract The spike (S) protein of SARS coronavirus (SARS-CoV) is responsible for viral binding with ACE2 molecules. Its receptor-binding motif (S-RBM) is located between residues 424 and 494, which folds into 2 anti-parallel β-sheets, β5 and β6. We have previously demonstrated that fragment 450–650 of the S protein (S450–650) is predominantly recognized by convalescent sera of SARS patients. The N-terminal 60 residues (450–510) of the S450–650 fragment covers the entire β6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the β6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450–650, mainly recognized the β6 strand-containing region. Unlike patient sera, however, the mouse antisera were unable to inhibit the infectivity of S protein-expressing (SARS-CoV-S) pseudovirus. Fusion protein between green fluorescence protein (GFP) and S450–650 (S450–650-GFP) was able to stain Vero E6 cells and deletion of the β6 fragment rendered the fusion product (S511–650-GFP) unable to do so. Similarly, recombinant S450–650, but not S511–650, was able to block the infection of Vero E6 cells by the SARS-CoV-S pseudovirus. Co-precipitation experiments confirmed that S450–650 was able to specifically bind with ACE2 molecules in lysate of Vero E6 cells. However, the ability of S450–510, either alone or in fusion with GFP, to bind with ACE2 was significantly poorer compared with S450–650. Our data suggest a possibility that, although the β6 strand alone is able to bind with ACE2 with relatively high affinity, residues outside the S-RBM could also assist the receptor binding of SARS-CoV-S protein.


title: Early events of SARS coronavirus infection in vero cells

text: An isolate from a patient in the recent severe acute respiratory syndrome (SARS) outbreak in Singapore was used to infect Vero E6 cells. This study concentrated on the first 30 min of infection. It was discovered that the SARS coronavirus attached, entered, and uncoated the nucleocapsids, all within a 30‐min period. At 5 min after infection, several virus particles lined the Vero cell plasma membrane. Virus particles were at various stages of fusion at the cell surface, since entry was not a synchronised process. After entry (10 and 15 min), spherical core particles moved into the cytoplasm within large vacuoles. Quite surprising at such early stages of infection (20 min), a virus‐induced change in the infected cells was evident. The induction of myelin‐like membrane whorls was obvious within the same vacuoles as the core particles. The significance of this virus‐induced change is unknown at this stage. By 25–30 min postinfection (p.i.), the spherical core particles appeared to be disassociating and, in their place, doughnut‐shaped electron‐dense structures were observed. These could be the virus genomes together with the helical nucleocapsids. They were no longer in large vacuoles but packaged into smaller vacuoles in the cytoplasm, and occasionally in small groups. J. Med. Virol. 71:323–331, 2003. © 2003 Wiley‐Liss, Inc.


title: Spiking the MERS-coronavirus receptor

text: A novel coronavirus, the Middle East respiratory syndrome coronavirus, recently emerged through zoonotic transmission, causing a severe lower respiratory tract infection in humans. In two recent papers, one published in Cell Research, the crystal structure of the viral receptor-binding domain in complex with the host CD26/dipeptidyl peptidase 4 receptor has now been characterized.


title: Simultaneous detection of DNA and RNA virus species involved in bovine respiratory disease by PCR-free rapid tagmentation-based library preparation and MinION nanopore sequencing

text: The Oxford Nanopore MinION Mk1B is a portable 90 g device that sequences DNA directly at 450 bases/second generating sequence reads in excess of 400 kb. Recent improvements in error rate and speed of library preparation mean that this device has considerable potential for rapid molecular bovine pathogen diagnostics. We tested the MinION for rapid untargeted detection of viral pathogens associated with bovine respiratory disease (BRD), an economically important disease often involving primary infection of the lung by one or more of a number of DNA and/or RNA viruses. We combined three foetal lung cell cultures which were infected with either Bovine Respiratory Syncytial Virus (BRSV), Bovine Herpes Virus 1 (BoHV1) or Bovine Parainfluenza Virus 3 (BPI-3). BoHV1 is a DNA virus and BPI-3 and BRSV are RNA viruses. The cell cultures were treated with DNase and RNase to deplete bovine nucleic acid prior to viral nucleic acid extraction and double-stranded cDNA synthesis. Sequencing libraries were generated by PCR-free tagmentation in under 10 minutes and loaded onto a MinION sequencer. Approximately 7,000 sequencing reads were generated and analysed using high-throughput local BLAST against the NCBI nr/nt database. The top BLAST hit for 2,937 of these reads was identified as a virus. Of these, 2,926 (99.6%) were correctly identified either as BoHV1, BRSV or BPI-3.


title: Characterization of the potyviral HC-pro autoproteolytic cleavage site

text: Abstract The helper component-proteinase (HC-Pro) encoded by potyviruses functions to cleave the viral polyprotein by an autoproteolytic mechanism at the HC-Pro C-terminus. This protein belongs to a group of viral cysteine-type proteinases and has been shown previously to catalyze proteolysis between a Gly-Gly dipeptide. The amino acid sequence requirements surrounding the HC-Pro C-terminal cleavage site of the tobacco etch virus polyprotein have been investigated using site-directed mutagenesis and in vitro expression systems. A total of 51 polyprotein derivatives, each differing by the substitution of a single amino acid between the P5 and P2′ positions, were tested for autoproteolytic activity. Substitutions of Tyr (P4), Val (P2), Gly (P1), and Gly (P1′) were found to eliminate or nearly eliminate proteolysis. Substitutions of Thr (P5), Asn (P3), and Met (P2′), on the other hand, were permissive for proteolysis, although the apparent processing rates of some polyproteins containing these alterations were reduced. These results suggest that auto-recognition by HC-Pro involves the interaction of the enzymatic binding site with four amino acids surrounding the cleavage site. Comparison of the homologous sequences of five potyviral polyproteins revealed that the residues essential for processing are strictly conserved, whereas the nonessential residues are divergent. The relationship between HC-Pro and other viral and cellular cysteine-type proteinases is discussed.


title: Protein Histochemistry Using Coronaviral Spike Proteins: Studying Binding Profiles and Sialic Acid Requirements for Attachment to Tissues

text: Protein histochemistry is a tissue-based technique that enables the analysis of viral attachment patterns as well as the identification of specific viral and host determinants involved in the first step in the infection of a host cell by a virus. Applying recombinantly expressed spike proteins of infectious bronchitis virus onto formalin-fixed tissues allows us to profile the binding characteristics of these viral attachment proteins to tissues of various avian species. In particular, sialic acid-mediated tissue binding of spike proteins can be analyzed by pretreating tissues with various neuraminidases or by blocking the binding of the viral proteins with specific lectins. Our assay is particularly convenient to elucidate critical virus–host interactions for viruses for which infection models are limited.


title: Cancer, immune suppression and Coronavirus Disease-19 (COVID-19): Need to manage drug safety

text: Abstract The Coronavirus disease (COVID-19) pandemic is disrupting our health environment. As expected, studies highlighted the great susceptibility of cancer patients to COVID-19 and more severe complications, leading oncologists to deeply rethink patient cancer care. This review is dedicated to the optimization of care pathways and therapeutics in cancer patients during the pandemic and aims to discuss successive issues. First we focused on the international guidelines proposing adjustments and alternative options to cancer care in order to limit hospital admission and cytopenic treatment in cancer patients, most of whom are immunocompromised. In addition cancer patients are prone to polypharmacy, enhancing the risk of drug-related problems as adverse events and drug-drug interactions. Due to increased risk in case of COVID-19, we reported a comprehensive review of all the drug-related problems between COVID-19 and antineoplastics. Moreover, in the absence of approved drug against COVID-19, infected patients may be included in clinical trials evaluating new drugs with a lack of knowledge, particularly in cancer patients. Focusing on the several experimental drugs currently being evaluated, we set up an original data board helping oncologists and pharmacists to identify promptly drug-related problems between antineoplastics and experimental drugs. Finally additional and concrete recommendations are provided, supporting oncologists and pharmacists in their efforts to manage cancer patients and to optimize their treatments in this new era related to COVID-19.


title: Plant tissue distribution and chemical inactivation of six carnation viruses

text: Carnation mottle virus (CarMV), Carnation etched ring virus (CERV), Carnation vein mottle virus (CVMV), Carnation ringspot virus (CRSV), Carnation Italian ringspot virus (CIRV) and Carnation latent virus (CLV) are the most important viruses affecting carnation crops. All except CERV are RNA viruses. Viral RNA or DNA accumulation on root, stem, leaf, sepal, petal, stamen, pistil and ovary tissues of infected carnation or Saponaria vaccaria plants was analysed by non-isotopic molecular hybridisation. High-titres of CarMV, CRSV, CIRV, and CLV accumulated in all plant tissues whereas CERV and CVMV were irregularly distributed over the plant. High-titres of all viruses accumulated in leaf, petal, stamen, pistil, and ovary tissues, so leaves or petals are a good tissue for routine diagnosis. Six chemicals were evaluated for inactivation of all carnation viruses in infected extracts. Commercial bleach at 7% (v/v) or NaOH at 0.5% (w/v) was found to inactivate all viruses after 60 s treatment in a systemic S. vaccaria bioassay.


title: The IFN response in bat cells consists of canonical and non-canonical ISGs with unique temporal expression kinetics

text: Bats are reservoirs for a number of highly pathogenic zoonotic viruses, yet they remain relatively asymptomatic during infection. Whether this viral resistance is due to a unique innate immune system is unknown. An evolutionarily conserved feature of vertebrate antiviral immunity is the interferon (IFN) response, which triggers cellular defenses through interferon-stimulated gene (ISG) expression. While bats encode an intact IFN system, global ISG expression patterns in bat cells are not well characterized. Here, we used RNA-Seq to assess the transcriptional response to IFNα in cells derived from the bat Pteropus alecto (black flying fox). We show induction of more than 100 transcripts, most of which are canonical ISGs observed in other species. Kinetic gene profiling revealed that P. alecto ISGs fall into two unique temporal subclusters with similar early induction kinetics but distinct late-phase declines. In contrast to bat ISGs, human ISGs generally remained elevated for longer periods following IFN treatment, suggesting host-based differences in gene regulatory mechanisms. Notably, we also identified a small group of non-canonical bat ISGs, including an enzymatically active RNASEL that plays a role in controlling viral infection. These studies provide insight into the innate immune response of an important viral reservoir and lay a foundation for studies into the immunological features that may underlie unique virus-host relationship in bats. Bats are considered unique in their ability to resist disease caused by viruses that are often pathogenic in humans. While the nature of this viral resistance is unknown, genomic data suggest bat innate immune systems may be specialized in controlling these disease-causing viruses. A critical cell intrinsic antiviral defense system in vertebrates is the interferon response, which suppresses viral infection through induction of hundreds of interferon-stimulated genes (ISGs). In this study, we report the repertoire of ISGs and several unique features of ISG induction kinetics in bat cells. We also characterize induction and antiviral activity of bat RNASEL, which is induced by IFN in bat, but not human cells. These studies lay the foundation for discovery of potentially new antiviral mechanisms in bats, which may spur research into development of therapies to combat viral infection.


title: Preparing for an Influenza Pandemic: Hospital Acceptance Study of Filtering Facepiece Respirator Decontamination Using Ultraviolet Germicidal Irradiation

text: OBJECTIVES: Predictions estimate supplies of filtering facepiece respirators (FFRs) would be limited in the event of a severe influenza pandemic. Ultraviolet decontamination and reuse (UVDR) is a potential approach to mitigate an FFR shortage. A field study sought to understand healthcare workers’ perspectives and potential logistics issues related to implementation of UVDR methods for FFRs in hospitals. METHODS: Data were collected at three hospitals using a structured guide to conduct 19 individual interviews, 103 focus group interviews, and 285 individual surveys. Data were then evaluated using thematic analysis to reveal key themes. RESULTS: Data revealed noteworthy variation in FFR use across the sample, along with preferences and requirements for the use of UVDR, unit design, and FFR reuse. Based on a scale of 1 (low) to 10 (high), the mean perception of safety in a high mortality pandemic wearing no FFR was 1.25 of 10, wearing an FFR for an extended period without decontamination was 4.20 of 10, and using UVDR was 7.72 of 10. CONCLUSIONS: In addition to technical design and development, preparation and training will be essential to successful implementation of a UVDR program. Ultraviolet decontamination and reuse program design and implementation must account for actual clinical practice, compliance with regulations, and practical financial considerations to be successfully adopted so that it can mitigate potential FFR shortages in a pandemic.


title: Chapter 9 Business Continuity

text: Publisher Summary Business continuity planning (BCP) is a methodology for planning how to deal with unforeseen disruptions in financial business processes. These disruptions are expected to be of a large scale, such as the disaster of September 11, only 5% of the time; more often than not, disruptions are minor and on a much smaller scale. These smaller disruptions may be due to short-term power outages, fires, disk drive failures, or computer viruses. Regardless, the firm must make a plan based on a thorough analysis of all scenarios that may affect its normal working, and create and test contingency and recovery measures for such situations. This chapter discusses the salient parts of a BCP, which constitutes a strategy for recovering from sudden and unexpected disruption of critical business functionalities. Business continuity and disaster recovery became more important after the occurrence of the September 11, 2001, terrorist attacks in New York, which led to widespread failure in financial systems. Developing a business continuity and disaster recovery plan requires setting aside resources and treating the development of the plan as a project for the firm. Once the analysis has been conducted and strategies have been formulated, the plan must be tested using simulated emergency scenarios. These scenarios are also important when it comes to industry-wide testing, which has become more of a focus recently as firms in financial sectors across the world become more deeply interlinked. While it is crucial for each firm to have its own business continuity plan, it is just as important for the industry as a whole to protect its infrastructure, since systems of each firm are intricately linked together in an STP environment.


title: Intestinal gastrointestinal stromal tumor in a cat

text: A 12-year-old, 3.6-kg, spayed female domestic shorthaired cat had a 2-month history of anorexia and weight loss. Abdominal ultrasonography and computed tomography revealed an exophytic mass originating from the jejunum with very poor central and poor peripheral contrast enhancement. On day 14, surgical resection of the jejunum and mass with 5-cm margins and an end-to-end anastomosis were performed. Histopathological examination revealed the mass was a transmural, invasive cancer showing exophytic growth and originating from the small intestinal muscle layer. Immunohistochemical analysis of tumor cells revealed diffuse positivity for KIT protein and negativity for desmin and S-100. The mass was diagnosed as a gastrointestinal stromal tumor (GIST). Ultrasonographic findings indicated the tumor probably metastasized to the liver and omentum, as seen in humans and dogs. The owner rejected further treatment at the last visit on day 192. To our knowledge, this is the first report of intestinal tumor and metastasis in feline GIST and its imaging features.


title: Diagnosis of cytomegalovirus pneumonia by quantitative polymerase chain reaction using bronchial washing fluid from patients with hematologic malignancies

text: BACKGROUND: The incidence of cytomegalovirus (CMV) pneumonia is increasing in patients diagnosed with hematologic malignancies. The utility of CMV-DNA viral load measurement has not been standardized, and viral cut-off values have not been established. This study was designed to investigate the utility of CMV quantitative real-time PCR (qRT-PCR) using bronchial washing fluid. METHODS: We retrospectively reviewed the microbiologic and pathologic results of bronchial washing fluid and biopsy specimens in addition to the patients' clinical characteristics. RESULTS: A total of 565 CMV qRT-PCR assays were performed using bronchial washing fluid from patients with hematologic malignancies. Among them, 101 were positive for CMV by qRT-PCR; of these, 24 were diagnosed with CMV pneumonia and 70 with CMV infection, and 7 were excluded due to a diagnosis of invasive pulmonary aspergillosis rather than viral pneumonia. The median CMV load determined by qPCR was 1.8 × 10(5) copies/mL (3.6 10(3)-1.5 × 10(8)) in CMV pneumonia patients and 3.0 × 10(3) copies/mL (5.0 × 10(2)-1.1 × 10(5)) in those diagnosed with CMV infection (P < 0.01). Using the ROC curve, the optimal inflection points were 18,900 copies/mL (137,970 IU/mL) in post-bone marrow transplantation (BMT) patients, 316,415 copies/mL (2,309,825 IU/mL) in no-BMT patients and 28,774 copies/mL (210,054 IU/mL) in all patients. CONCLUSIONS: The CMV titers in bronchial washing fluid determined by qRT-PCR differed significantly between patients diagnosed with CMV pneumonia and those with CMV infection. The viral cut-off values in bronchial washing fluid were suggested for the diagnosis of CMV pneumonia, which were different depending on the BMT status.


docid: ebsohram

title: From the field to the lab — An European view on the global spread of PEDV

text: Abstract Porcine Epidemic Diarrhea Virus (PEDV) is a member of the genus Alphacoronavirus, in the family Coronaviridae, of the Nidovirales order and outbreaks of porcine epidemic diarrhoea (PED) were first recorded in England in the 1970s. Intriguingly the virus has since successfully made its way around the globe, while seemingly becoming extinct in parts of Europe before its recent return from Northern America. In this review we are re-evaluating the spread of PEDV, its biology and are looking at lessons learnt from both failure and success. While a new analysis of PEDV genomes demonstrates a wider heterogeneity of PEDV than previously anticipated with at least five rather than two genotypes, biological features of the virus and its replication also point towards credible control strategies to limit the impact of this re-emerging virus.


title: Vectored immunoprophylaxis: an emerging adjunct to traditional vaccination

text: The successful development of effective vaccines has been elusive for many of the world’s most important infectious diseases. Additionally, much of the population, such as the aged or immunocompromised, are unable to mount an effective immunologic response for existing vaccines. Vectored Immunoprophylaxis (VIP) is a novel approach designed to address these challenges. Rather than utilizing an antigen to trigger a response from the host’s immune system as is normally done with traditional vaccines, VIP genetically engineers the production of tailored antibodies from non-hematopoietic cells, bypassing the humoral immune system. Direct administration of genes encoding for neutralizing antibodies has proven to be effective in both preventing and treating several infectious diseases in animal models. While, a significant amount of work has focused on HIV, including an ongoing clinical trial, the approach has also been shown to be effective for malaria, dengue, hepatitis C, influenza, and more. In addition to presenting itself as a potentially efficient approach to solving long-standing vaccine challenges, the approach may be the best, if not only, method to vaccinate immunocompromised individuals. Many issues still need to be addressed, including which tissue(s) makes the most suitable platform, which vector(s) are most efficient at transducing the platform tissue used to secrete the antibodies, and what are the long-term effects of such a treatment. Here we provide a brief overview of this approach, and its potential application in treating some of the world’s most intractable infectious diseases.


title: Epithelial injury and interstitial fibrosis in the proximal alveolar regions of rats chronically exposed to a simulated pattern of urban ambient ozone

text: Abstract Electron microcopic morphometry was used to study the development of lung injury during and after chronic (78 weeks) exposure to a pattern of ozone (O3) designed to simulate high urban ambient concentrations that occur in some environments. The daily exposure regimen consisted of a 13-hr background of 0.06 ppm, an exposure peak that rose from 0.06 to 0.25 ppm, and returned to the background level over a 9-hr period, and 2-hr downtime for maintenance. Rats were exposed for 1, 3, 13, and 78 weeks. Additional groups of rats exposed for 13 or 78 weeks were allowed to recover in filtered clean air for 6 or 17 weeks, respectively. Rats exposed to filtered air for the same lengths of time were used as controls. Samples from proximal alveolar regions and terminal bronchioles were obtained by microdissection. Analysis of the proximal alveolar region revealed a biphasic response. Acute tissue reactions after 1 week of exposure included epithelial inflammation, interstitial edema, interstitial cell hypertrophy, and influx of macrophages. These responses subsided after 3 weeks of exposure. Progressive epithelial and interstitial tissue responses developed with prolonged exposure and included epithelial hyperplasia, fibroblast proliferation, and interstitial matrix accumulation. The epithelial responses involved both type I and type II epithelial cells. Alveolar type I cells increased in number, became thicker, and covered a smaller average surface area. These changes persisted throughout the entire exposure and did not change during the recovery pefiod, indicating the sensitivity of these cells to injury. The main response of type II epithelial cells was cell proliferation. The accumulation of interstitial matrix after chronic exposure consisted of deposition of both increased amounts of basement membrane and collagen fibers. Interstitial matrix accumulation underwent partial recovery during follow-up periods in air; however, the thickening of the basement membrane did not resolve. Analysis of terminal bronchioles showed that short-term exposure to O3 caused a loss of ciliated cells and differentiation of preciliated and Clara cells. The bronchiolar cell population stabilized on continued exposure; however, chronic exposure resulted in structural changes, suggesting injury to both ciliated and Clara cells. We conclude that chronic exposure to low levels of O3 causes epithelial inflammation and interstitial fibrosis in the proximal alveolar region and bronchiolar epithelial cell injury.


title: Sars‐CoV‐2 infection: the same virus can cause different cutaneous manifestations

text: The article of Galván Casas et al. describing the cutaneous manifestations occurring during the COVID‐19 pandemic(1) prompted us to make some observations. Informations regarding the prevalence of enanthems, their clinical features and the association with exanthem patterns and disease severity are lacking. In the absence of a clearly defined temporal connection, the relationship between exanthems and SARS‐CoV‐2 infection is at times unclear, as in the case of urticarial/maculopapular eruptions in patients undergoing treatment with multiple drugs.


title: Molecular epidemiology of pathogenic Leptospira spp. in the straw-colored fruit bat (Eidolon helvum) migrating to Zambia from the Democratic Republic of Congo

text: Abstract The role played by bats as a potential source of transmission of Leptospira spp. to humans is poorly understood, despite various pathogenic Leptospira spp. being identified in these mammals. Here, we investigated the prevalence and diversity of pathogenic Leptospira spp. that infect the straw-colored fruit bat (Eidolon helvum). We captured this bat species, which is widely distributed in Africa, in Zambia during 2008–2013. We detected the flagellin B gene (flaB) from pathogenic Leptospira spp. in kidney samples from 79 of 529 E. helvum (14.9%) bats. Phylogenetic analysis of 70 flaB fragments amplified from E. helvum samples and previously reported sequences, revealed that 12 of the fragments grouped with Leptospira borgpetersenii and Leptospira kirschneri; however, the remaining 58 flaB fragments appeared not to be associated with any reported species. Additionally, the 16S ribosomal RNA gene (rrs) amplified from 27 randomly chosen flaB-positive samples was compared with previously reported sequences, including bat-derived Leptospira spp. All 27 rrs fragments clustered into a pathogenic group. Eight fragments were located in unique branches, the other 19 fragments were closely related to Leptospira spp. detected in bats. These results show that rrs sequences in bats are genetically related to each other without regional variation, suggesting that Leptospira are evolutionarily well-adapted to bats and have uniquely evolved in the bat population. Our study indicates that pathogenic Leptospira spp. in E. helvum in Zambia have unique genotypes.


title: Management of neurological care during the COVID-19 pandemic

text: Abstract Introduction The COVID-19 epidemic has led to the need for unprecedented decisions to be made to maintain the provision of neurological care. This article addresses operational decision-making during the epidemic. Development We report the measures taken, including the preparation of a functional reorganisation plan, strategies for hospitalisation and emergency management, the use of telephone consultations to maintain neurological care, provision of care at a unit outside the hospital for priority patients, decisions about complementary testing and periodic in-hospital treatments, and the use of a specific telephone service to prioritise patients with epileptic seizures. Conclusion Despite the situation of confinement, neurology departments must continue to provide patient care through different means of operation. Like all elements of management, these must be evaluated.


title: Metagenomic Analysis of the Ferret Fecal Viral Flora

text: Ferrets are widely used as a small animal model for a number of viral infections, including influenza A virus and SARS coronavirus. To further analyze the microbiological status of ferrets, their fecal viral flora was studied using a metagenomics approach. Novel viruses from the families Picorna-, Papilloma-, and Anelloviridae as well as known viruses from the families Astro-, Corona-, Parvo-, and Hepeviridae were identified in different ferret cohorts. Ferret kobu- and hepatitis E virus were mainly present in human household ferrets, whereas coronaviruses were found both in household as well as farm ferrets. Our studies illuminate the viral diversity found in ferrets and provide tools to prescreen for newly identified viruses that potentially could influence disease outcome of experimental virus infections in ferrets.


title: Nicotine and smoking in the COVID-19 era

text: Introduction: The knowledge regarding the demographic characteristics of patients with Covid-19 and risk factors distribution is still evolving. Considering the role of cigarette smoking in the pathogenesis of lung diseases and the effect of nicotine on expression of the entry receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it is important to determine the implications of smoking in COVID-19. Methods: In this brief report, by using the published articles in the literature, we aimed to compare the reported prevalence of smoking in patients with COVID-19 to the prevalence of smoking in the general population of the corresponding report. Binomial tests were conducted and a P value of less than 0.05 was considered statistically significant. Results: Among the screened papers, we found 12 peer-reviewed articles in which epidemiological characteristics of COVID-19 patients, including smoking status, were stated. Based on the descriptive reports of characteristics of COVID-19 patients, we observed a significantly lower proportion of COVID-19 patients with smoking history compared to what is expected, given the population average for each study’s geographic area. Conclusion: This analysis of available data showed a lower prevalence of smoking in COVID-19 patients in comparison to the regional average. Considering the limitations of the study, the results should be interpreted with great caution and be viewed just as a preliminary report to motivate related basic and clinical researches.


docid: nfdsomaz

title: Changes in mortalities and hospital admissions associated with holidays and respiratory illness: implications for medical services

text: Rationale and objectives To see whether net mortalities increase during and after reductions in medical services, either at average weekends, or at Christmas when pressure from illness is unusually high. Methods (1) Paired t‐tests to compare mean daily deaths and hospital admissions during and after weekends (Saturday–Tuesday) with means for the week, in south‐east England; (2) Linear regressions to see whether trends of daily deaths change when admissions are reduced at Christmas. Results Neither mean daily all‐cause, respiratory or ischaemic heart deaths exceeded weekly averages during weekends, or during Saturday–Monday or Saturday–Tuesday, despite falls in daily elective and daily emergency hospital admissions at weekends that averaged 61–72% and 14–22%, respectively. During 19–24 December, daily deaths were above annual means, respiratory deaths by 49% (29, 1–58), but elective admissions fell and although emergency admissions tended to rise, total admissions rose only for respiratory disease, and only by 33% (376, −47 to 799). On Christmas Day (25 December), even emergency admissions fell sharply below previous trends, respiratory emergency admissions by 18% (P < 0.01). Respiratory deaths alone then immediately increased (P < 0.01) above trend, by 5.9% (5.8 deaths/day) on 26 December and by 12.9% (12.9) on 27 December. Conclusions No adverse effect on mortality was apparent within 2 days from reduction in medical services at weekends. However, respiratory deaths accelerated sharply after reduction in elective and emergency admissions at Christmas, when rates of infection and mortality from respiratory disease were high. Implications for medical services during respiratory epidemics are discussed.


title: Comparison of virological profiles of respiratory syncytial virus and rhinovirus in acute lower tract respiratory infections in very young Chilean infants, according to their clinical outcome

text: Abstract Background Respiratory syncytial virus (RSV) and rhinovirus (HRV) are the main cause of acute lower respiratory tract infections (ALRTIs) in infants. Viral and host-related risk factors for severe disease have also not been clearly established. Objective To assess whether certain viral features of RSV and, or HRV are associated with severe ALRTI. Study design RSV and HRV were studied in nasopharyngeal samples of infants by immunofluorescence, Luminex® and/or real-time RT-PCR assays. Quantitation and genotyping of RSV and HRV by PCR were done. Results Of 124 virus positive specimens, 74 (59.7%) had RSV; 22 (17.7%) HRV and 28 (22.6%) RSV-HRV co-infection. Hospitalization was required in 57/74 RSV infants (77.0%); in 10/22 HRV cases (45.5%) (p =0.006) and in 15/28 co-infected by both viruses (53.6%) (p =0.003). Severe cases were 33/74 (44.6%) RSV infections, 2/22 HRV cases (9.1%), (p <0.002) and 6/28 (21.4%) patients co-infected by RSV–HRV (p <0.026). Three genotypes (NA1, B7, B9) of RSV circulated during the study. In 33 severe infants, NA1 was detected in 19 cases (57.6%); B7 in 13 (39.4%) and B9 in 1 (3.0%) (p <0.01; OR=10.0). RSV loads were similar between outpatients and hospitalized infants (p =0.7) and among different severities (p =0.7). NA1 loads were higher than other strains (p =0.049). Three geno-groups of HRV circulated homogeneously. Conclusion In very young infants, RSV cause more severe disease than HRV. Co-infection does not increase the severity of illness. NA1 RSV genotype was associated with major frequency of hospitalization, severe respiratory disease and higher viral load.


docid: amjrzocv

title: Both STING and MAVS Fish Orthologs Contribute to the Induction of Interferon Mediated by RIG-I

text: Viral infections are detected in most cases by the host innate immune system through pattern-recognition receptors (PRR), the sensors for pathogen-associated molecular patterns (PAMPs), which induce the production of cytokines, such as type I interferons (IFN). Recent identification in mammalian and teleost fish of cytoplasmic viral RNA sensors, RIG-I-like receptors (RLRs), and their mitochondrial adaptor: the mitochondrial antiviral signaling (MAVS) protein, also called IPS-1, highlight their important role in the induction of IFN at the early stage of a virus infection. More recently, an endoplasmic reticulum (ER) adaptor: the stimulator of interferon genes (STING) protein, also called MITA, ERIS and MPYS, has been shown to play a pivotal role in response to both non-self-cytosolic RNA and dsDNA. In this study, we cloned STING cDNAs from zebrafish and showed that it was an ortholog to mammalian STING. We demonstrated that overexpression of this ER protein in fish cells led to a constitutive induction of IFN and interferon-stimulated genes (ISGs). STING-overexpressing cells were almost fully protected against RNA virus infection with a strong inhibition of both DNA and RNA virus replication. In addition, we found that together with MAVS, STING was an important player in the RIG-I IFN-inducing pathway. This report provides the demonstration that teleost fish possess a functional RLR pathway in which MAVS and STING are downstream signaling molecules of RIG-I. The Sequences presented in this article have been submitted to GenBank under accession numbers: Zebrafish STING (HE856619); EPC STING (HE856620); EPC IRF3 (HE856621); EPC IFN promoter (HE856618).


title: Emerging and experimental treatments for COVID-19 and drug interactions with psychotropic agents

text: As yet, no agents have been approved for the treatment of COVID-19, although several experimental drugs are being used off licence. These may have serious adverse effects and potential drug interactions with psychotropic agents. We reviewed the common agents being used across the world for the treatment of COVID-19 and investigated their drug interaction potential with psychotropic agents using several drug interaction databases and resources. A preliminary search identified the following drugs as being used to treat COVID-19 symptoms: atazanavir (ATV), azithromycin (AZI), chloroquine (CLQ)/hydroxychloroquine (HCLQ), dipyridamole, famotidine (FAM), favipiravir, lopinavir/ritonavir (LPV/r), nitazoxanide, remdesivir, ribavirin and tocilizumab. Many serious adverse effects and potential drug interactions with psychotropic agents were identified. The most problematic agents were found to be ATV, AZI, CLQ, HCLQ, FAM and LPV/r in terms of both pharmacokinetic as well as serious pharmacodynamic drug interactions, including QTc prolongation and neutropenia. Significant caution should be exercised if using any of the medications being trialled for the treatment of COVID-19 until robust clinical trial data are available. An even higher threshold of vigilance should be maintained for patients with pre-existing conditions and older adults due to added toxicity and drug interactions, especially with psychotropic agents.


How to cite this article: Elangovan A. QTc Prolongation in the Critically Ill: Tread with Caution! Indian J Crit Care Med 2020;24(4):220–221.


title: Inhibition of canonical WNT signaling attenuates human leiomyoma cell growth

text: OBJECTIVE: Dysregulation of WNT signaling plays a central role in tumor cell growth and progression. Our goal was to assess the effect of three WNT/β-catenin pathway inhibitors, Inhibitor of β-Catenin And TCF4 (ICAT), niclosamide, and XAV939 on the proliferation of primary cultures of human uterine leiomyoma cells. DESIGN: Prospective study of human leiomyoma cells obtained from myomectomy or hysterectomy. SETTING: University research laboratory. PATIENT(S): Women (n=38) aged 27–53 years undergoing surgery. INTERVENTION(S): Adenoviral ICAT overexpression or treatment with varying concentrations of niclosamide or XAV939. MAIN OUTCOME MEASURE(S): Cell proliferation, cell death, WNT/β-catenin target gene expression or reporter gene regulation, β-catenin levels and cellular localization. RESULT(S): ICAT, niclosamide, or XAV939 inhibit WNT/β-catenin pathway activation and exert anti-proliferative effects in primary cultures of human leiomyoma cells. CONCLUSION(S): Three WNT/β-catenin pathway inhibitors specifically block human leiomyoma growth and proliferation, suggesting that the canonical WNT pathway may be a potential therapeutic target for the treatment of uterine leiomyoma. Our findings provide rationale for further preclinical and clinical evaluation of ICAT, niclosamide, and XAV939 as candidate anti-tumor agents for uterine leiomyoma.


title: The V-type H+-ATPase in vesicular trafficking: targeting, regulation and function

text: Vacuolar-type H+-ATPase (V-ATPase)-driven proton pumping and organellar acidification is essential for vesicular trafficking along both the exocytotic and endocytotic pathways of eukaryotic cells. Deficient function of V-ATPase and defects of vesicular acidification have been recently recognized as important mechanisms in a variety of human diseases and are emerging as potential therapeutic targets. In the past few years, significant progress has been made in our understanding of function, regulation, and the cell biological role of V-ATPase. Here, we will review these studies with emphasis on novel direct roles of V-ATPase in the regulation of vesicular trafficking events.


title: Effect of non-pharmaceutical interventions for containing the COVID-19 outbreak in China

text: BACKGROUND: The COVID-19 outbreak containment strategies in China based on non-pharmaceutical interventions (NPIs) appear to be effective. Quantitative research is still needed however to assess the efficacy of different candidate NPIs and their timings to guide ongoing and future responses to epidemics of this emerging disease across the World. METHODS: We built a travel network-based susceptible-exposed-infectious-removed (SEIR) model to simulate the outbreak across cities in mainland China. We used epidemiological parameters estimated for the early stage of outbreak in Wuhan to parameterise the transmission before NPIs were implemented. To quantify the relative effect of various NPIs, daily changes of delay from illness onset to the first reported case in each county were used as a proxy for the improvement of case identification and isolation across the outbreak. Historical and near-real time human movement data, obtained from Baidu location-based service, were used to derive the intensity of travel restrictions and contact reductions across China. The model and outputs were validated using daily reported case numbers, with a series of sensitivity analyses conducted. RESULTS: We estimated that there were a total of 114,325 COVID-19 cases (interquartile range [IQR] 76,776 – 164,576) in mainland China as of February 29, 2020, and these were highly correlated (p<0.001, R(2)=0.86) with reported incidence. Without NPIs, the number of COVID-19 cases would likely have shown a 67-fold increase (IQR: 44 – 94), with the effectiveness of different interventions varying. The early detection and isolation of cases was estimated to prevent more infections than travel restrictions and contact reductions, but integrated NPIs would achieve the strongest and most rapid effect. If NPIs could have been conducted one week, two weeks, or three weeks earlier in China, cases could have been reduced by 66%, 86%, and 95%, respectively, together with significantly reducing the number of affected areas. However, if NPIs were conducted one week, two weeks, or three weeks later, the number of cases could have shown a 3-fold, 7-fold, and 18-fold increase across China, respectively. Results also suggest that the social distancing intervention should be continued for the next few months in China to prevent case numbers increasing again after travel restrictions were lifted on February 17, 2020. CONCLUSION: The NPIs deployed in China appear to be effectively containing the COVID-19 outbreak, but the efficacy of the different interventions varied, with the early case detection and contact reduction being the most effective. Moreover, deploying the NPIs early is also important to prevent further spread. Early and integrated NPI strategies should be prepared, adopted and adjusted to minimize health, social and economic impacts in affected regions around the World.


title: Analysis of Imported Cases of COVID-19 in Taiwan: A Nationwide Study

text: In the early stages of the 2019 novel coronavirus disease (COVID-19) pandemic, containment of disease importation from epidemic areas was essential for outbreak control. This study is based on publicly accessible data on confirmed COVID-19 cases in Taiwan extracted from the Taiwan Centers for Disease Control website. We analysed the characteristics, infection source, symptom presentation, and route of identification of the 321 imported cases that were identified from 21 January to 6 April 2020. They were mostly returned Taiwanese citizens who had travelled to one or more of 37 countries for tourism, business, work, or study. Half of these cases developed symptoms before arrival, most of the remainder developed symptoms 1–13 days (mean 4.0 days) after arrival, and 3.4% never developed symptoms. Three-quarters of the cases had respiratory symptoms, 44.9% had fever, 13.1% lost smell or taste, and 7.2% had diarrhoea. Body temperature and symptom screening at airports identified 32.7% of the cases. Of the remainder, 27.7% were identified during home quarantining, 16.2% were identified via contact tracing, and 23.4% were reported by hospitals. Under the strict enforcement of these measures, the incidence of locally acquired COVID-19 cases in Taiwan remains sporadic. In conclusion, proactive border control measures are effective for preventing community transmission of this disease.


title: Cytomegalovirus-Induced Adrenal Insufficiency in a Renal Transplant Recipient

text: Abstract Cytomegalovirus (CMV) is an important pathogen in organ-transplant recipients. There have been frequent reports of CMV-induced adrenal insufficiency in patients with human immunodeficiency virus infection. Herein, we report CMV-induced renal insufficiency in a renal transplant recipient. A 24-year-old woman had gradual onset of weakness, anorexia, nausea, hypotension, and skin hyperpigmentation at 5 months after renal transplantation. The immunosuppression regimen included cyclosporine, mycophenolate mofetil, and corticosteroid (prednisolone, 5 mg/d). Recent history included acute CMV infection, which was treated with ganciclovir. Basal serum cortisol concentration was 4 μg/dL, and stimulated serum cortisol concentration was less than 10 μg/dL. All clinical signs and symptoms and hypotension gradually improved after the oral prednisolone dose was increased to 10 mg/d. Clinicians must be aware of the possibility of CMV-induced adrenal insufficiency in renal transplant recipients. The condition may be symptomatic despite low-dose prednisolone therapy.


title: Expression of SARS-CoV-2 Entry Molecules ACE2 and TMPRSS2 in the Gut of Patients With IBD

text: BACKGROUND: Patients with inflammatory bowel disease (IBD) have intestinal inflammation and are treated with immune-modulating medications. In the face of the coronavirus disease-19 pandemic, we do not know whether patients with IBD will be more susceptible to infection or disease. We hypothesized that the viral entry molecules angiotensin I converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) are expressed in the intestine. We further hypothesized that their expression could be affected by inflammation or medication usage. METHODS: We examined the expression of Ace2 and Tmprss2 by quantitative polymerase chain reacion in animal models of IBD. Publicly available data from organoids and mucosal biopsies from patients with IBD were examined for expression of ACE2 and TMPRSS2. We conducted RNA sequencing for CD11b-enriched cells and peripheral and lamina propria T-cells from well-annotated patient samples. RESULTS: The molecules ACE2 and TMPRSS2 were abundantly expressed in the ileum and colon and had high expression in intestinal epithelial cells. In animal models, inflammation led to downregulation of epithelial Ace2. Expression of ACE2 and TMPRSS2 was not increased in samples from patients with compared with those of control patients. In CD11b-enriched cells but not T-cells, the level of expression of ACE2 and TMPRSS2 in the mucosa was comparable to other functional mucosal genes and was not affected by inflammation. Anti-tumor necrosis factor drugs, vedolizumab, ustekinumab, and steroids were linked to significantly lower expression of ACE2 in CD11b-enriched cells. CONCLUSIONS: The viral entry molecules ACE2 and TMPRSS2 are expressed in the ileum and colon. Patients with IBD do not have higher expression during inflammation; medical therapy is associated with lower levels of ACE2. These data provide reassurance for patients with IBD.


Lessons From the Coronavirus Crisis for European Integration


title: A vulnerability-based approach to human-mobility reduction for countering COVID-19 transmission in London while considering local air quality

text: Abstract An ecologic analysis was conducted to explore the correlation between air pollution, and COVID-19 cases and fatality rates in London. The analysis demonstrated a strong correlation (R2 > 0.7) between increment in air pollution and an increase in the risk of COVID-19 transmission within London boroughs. Particularly, strong correlations (R2 > 0.72) between the risk of COVID-19 fatality and nitrogen dioxide and particulate matter pollution concentrations were found. Although this study assumed the same level of air pollution across a particular London borough, it demonstrates the possibility to employ air pollution as an indicator to rapidly identify the city's vulnerable regions. Such an approach can inform the decisions to suspend or reduce the operation of different public transport modes within a city. The methodology and learnings from the study can thus aid in public transport's response to COVID-19 outbreak by adopting different levels of human-mobility reduction strategies based on the vulnerability of a given region.


title: Water and concentrate intake, weight gain and duration of diarrhea in young suckling calves on different diets

text: Abstract Diarrheic calves are fed with milk or milk replacer and oral rehydration solutions (ORS) to ensure energy and electrolyte supply. An easy and time-saving method is the preparation of ORS in milk. As milk-based ORS are hypertonic solutions administration of them may trigger thirst. Therefore, we hypothesized that restrictively fed calves receiving ORS prepared in milk had a higher water intake than restrictively and ad libitum fed calves receiving ORS prepared in water during diarrheic episodes. The daily water intake was measured in 100 individually-housed Holstein Friesian calves from day 2 to 21 of life. One group of the calves was fed with restrictive amounts of milk, the other group got milk ad libitum by an automated milk feeder. Nearly all calves spontaneously developed diarrhea within the observation period from day 2 to 21 of life. In cases of diarrhea the restrictively-fed calves received ORS prepared in milk or ORS prepared in water two hours after their milk meal, whereas the ad libitum-fed calves only got ORS prepared in water. All calves had ad libitum access to water. The daily intake of water, milk, and ORS and weight gain during diarrheic episode were determined. Data were expressed as arithmetic means (±standard deviation) and analyzed by using a one-way ANOVA or repeated-measures ANOVA. From day 2 to 21 of life calves fed with restrictive amounts of milk had higher water intakes related to the total dry matter intake (DMI) with 1.6L/kg of total DMI than ad libitum-fed calves (0.9L/kg of total DMI) per day. In cases of diarrhea water intake increased in all feeding groups. The calves receiving milk-based ORS had the highest water intake with 1.7L/d during the period of diarrhea compared to the calves received ORS prepared in water. Moreover, the calves fed ORS in milk showed with 4.6L/d the highest daily ORS intake. There were no differences in the duration of diarrhea or the daily weight gain during period of diarrhea between the feeding regimens. Therefore, it can be concluded that all feeding regimens were suitable in the treatment of calf diarrhea. The simplest method to treat calves suffering from diarrhea is the preparation of ORS in milk, but then ad libitum availability of water is absolutely necessary. Moreover, calves drink considerable amounts of water within the first 3 weeks of life and therefore should be provided with water for animal welfare reasons.


title: The revolution of the biology of the genome

text: Sequence data of entire eukaryotic genomes and their detailed comparison have provided new evidence on genome evolution. The major mechanisms involved in the increase of genome sizes are polyploidization and gene duplication. Subsequent gene silencing or mutations, preferentially in regulatory sequences of genes, modify the genome and permit the development of genes with new properties. Mechanisms such as lateral gene transfer, exon shuffling or the creation of new genes by transposition contribute to the evolution of a genome, but remain of relatively restricted relevance. Mechanisms to decrease genome sizes and, in particular, to remove specific DNA sequences, such as blocks of satellite DNAs, appear to involve the action of RNA interference (RNAi). RNAi mechanisms have been proven to be involved in chromatin packaging related with gene inactivation as well as in DNA excision during the macronucleus development in ciliates.


title: Middle East Respiratory Syndrome in 3 Persons, South Korea, 2015

text: In May 2015, Middle East respiratory syndrome coronavirus infection was laboratory confirmed in South Korea. Patients were a man who had visited the Middle East, his wife, and a man who shared a hospital room with the index patient. Rapid laboratory confirmation will facilitate subsequent prevention and control for imported cases.


title: Human lung ex vivo infection models

text: Pneumonia is counted among the leading causes of death worldwide. Viruses, bacteria and pathogen-related molecules interact with cells present in the human alveolus by numerous, yet poorly understood ways. Traditional cell culture models little reflect the cellular composition, matrix complexity and three-dimensional architecture of the human lung. Integrative animal models suffer from species differences, which are of particular importance for the investigation of zoonotic lung diseases. The use of cultured ex vivo infected human lung tissue may overcome some of these limitations and complement traditional models. The present review gives an overview of common bacterial lung infections, such as pneumococcal infection and of widely neglected pathogens modeled in ex vivo infected lung tissue. The role of ex vivo infected lung tissue for the investigation of emerging viral zoonosis including influenza A virus and Middle East respiratory syndrome coronavirus is discussed. Finally, further directions for the elaboration of such models are revealed. Overall, the introduced models represent meaningful and robust methods to investigate principles of pathogen-host interaction in original human lung tissue.


Familiarity breeds contempt


title: Tissue-Resident NK Cells: Development, Maturation, and Clinical Relevance

text: Natural killer (NK) cells belong to type 1 innate lymphoid cells (ILC1) and are essential in killing infected or transformed cells. NK cells mediate their effector functions using non-clonotypic germ-line-encoded activation receptors. The utilization of non-polymorphic and conserved activating receptors promoted the conceptual dogma that NK cells are homogeneous with limited but focused immune functions. However, emerging studies reveal that NK cells are highly heterogeneous with divergent immune functions. A distinct combination of several activation and inhibitory receptors form a diverse array of NK cell subsets in both humans and mice. Importantly, one of the central factors that determine NK cell heterogeneity and their divergent functions is their tissue residency. Decades of studies provided strong support that NK cells develop in the bone marrow. However, evolving evidence supports the notion that NK cells also develop and differentiate in tissues. Here, we summarize the molecular basis, phenotypic signatures, and functions of tissue-resident NK cells and compare them with conventional NK cells.


title: Distinct CD4 T-cell effects on primary versus recall CD8 T-cell responses during viral encephalomyelitis

text: CD4 T-cell help is not a universal requirement for effective primary CD8 T cells but is essential to generate memory CD8 T cells capable of recall responses. This study examined how CD4 T cells affect primary and secondary anti-viral CD8 T-cell responses within the central nervous system (CNS) during encephalomyelitis induced by sublethal gliatropic coronavirus. CD4 T-cell depletion before infection did not impair peripheral expansion, interferon-γ production, CNS recruitment or initial CNS effector capacity of virus-specific CD8 T cells ex vivo. Nevertheless, impaired virus control in the absence of CD4 T cells was associated with gradually diminished CNS CD8 T-cell interferon-γ production. Furthermore, within the CD8 T-cell population short-lived effector cells were increased and memory precursor effector cells were significantly decreased, consistent with higher T-cell turnover. Transfer of memory CD8 T cells to reduce viral load in CD4-depleted mice reverted the recipient CNS CD8 T-cell phenotype to that in wild-type control mice. However, memory CD8 T cells primed without CD4 T cells and transferred into infected CD4-sufficient recipients expanded less efficiently and were not sustained in the CNS, contrasting with their helped counterparts. These data suggest that CD4 T cells are dispensable for initial expansion, CNS recruitment and differentiation of primary resident memory CD8 T cells as long as the duration of antigen exposure is limited. By contrast, CD4 T cells are essential to prolong primary CD8 T-cell function in the CNS and imprint memory CD8 T cells for recall responses.


title: Astrovirus infections in humans and animals – Molecular biology, genetic diversity, and interspecies transmissions

text: Abstract Astroviruses are small, non-enveloped, positive sense, single-stranded RNA viruses first identified in 1975 in children suffering from diarrhea and then described in a wide variety of animals. To date, the list of animal species susceptible to astrovirus infection has expanded to 22 animal species or families, including domestic, synantropic and wild animals, avian, and mammalian species in the terrestrial and aquatic environments. Astrovirus infections are considered among the most common cause of gastroenteritis in children, second only to rotavirus infections, but in animals their association with enteric diseases is not well documented, with the exception of turkey and mink astrovirus infection. Genetic variability has been described in almost all astrovirus species sufficiently examined infecting mammals and birds; however, antigenic variability has been demonstrated for human astroviruses but is far less investigated in animal viruses. Interestingly, there is an increasing evidence of recombination events occurring in astroviruses, which contributes to increase the genetic variability of this group of viruses. A wide variety of species infected, the evident virus genetic diversity and the occurrence of recombination events indicate or imply either cross-species transmission and subsequent virus adaptation to new hosts or the co-infection of the same host with different astroviruses. This can also favor the emergence of novel astroviruses infecting animals or with a zoonotic potential. After more than 30years from their first description in humans, there are many exciting streams of research to be explored and intriguing questions that remain to be answered about the relatively under-studied Astroviridae family. In the present work, we will review the existing knowledge concerning astrovirus infections in humans and animals, with particular focus on the molecular biology, interspecies transmission and zoonotic potential of this group of viruses.


title: Fully human broadly neutralizing monoclonal antibodies against influenza A viruses generated from the memory B cells of a 2009 pandemic H1N1 influenza vaccine recipient

text: Abstract Whether the 2009 pandemic H1N1 influenza vaccine can induce heterosubtypic cross-protective anti-hemagglutinin (HA) neutralizing antibodies is an important issue. We obtained a panel of fully human monoclonal antibodies from the memory B cells of a 2009 pandemic H1N1 influenza vaccine recipient. Most of the monoclonal antibodies targeted the HA protein but not the HA1 fragment. Among the analyzed antibodies, seven mAbs exhibited neutralizing activity against several influenza A viruses of different subtypes. The conserved linear epitope targeted by the neutralizing mAbs (FIEGGWTGMVDGWYGYHH) is part of the fusion peptide on HA2. Our work suggests that a heterosubtypic neutralizing antibody response primarily targeting the HA stem region exists in recipients of the 2009 pandemic H1N1 influenza vaccine. The HA stem region contains various conserved neutralizing epitopes with the fusion peptide as an important one. This work may aid in the design of a universal influenza A virus vaccine.


title: The SARS Coronavirus E Protein Interacts with PALS1 and Alters Tight Junction Formation and Epithelial Morphogenesis

text: Intercellular tight junctions define epithelial apicobasal polarity and form a physical fence which protects underlying tissues from pathogen invasions. PALS1, a tight junction-associated protein, is a member of the CRUMBS3-PALS1-PATJ polarity complex, which is crucial for the establishment and maintenance of epithelial polarity in mammals. Here we report that the carboxy-terminal domain of the SARS-CoV E small envelope protein (E) binds to human PALS1. Using coimmunoprecipitation and pull-down assays, we show that E interacts with PALS1 in mammalian cells and further demonstrate that the last four carboxy-terminal amino acids of E form a novel PDZ-binding motif that binds to PALS1 PDZ domain. PALS1 redistributes to the ERGIC/Golgi region, where E accumulates, in SARS-CoV–infected Vero E6 cells. Ectopic expression of E in MDCKII epithelial cells significantly alters cyst morphogenesis and, furthermore, delays formation of tight junctions, affects polarity, and modifies the subcellular distribution of PALS1, in a PDZ-binding motif-dependent manner. We speculate that hijacking of PALS1 by SARS-CoV E plays a determinant role in the disruption of the lung epithelium in SARS patients.


title: Assessing the role of long-distance translocation and spatial heterogeneity in the raccoon rabies epidemic in Connecticut

text: Abstract Spatial heterogeneity and long-distance translocation (LDT) play important roles in the spatio-temporal dynamics and management of emerging infectious diseases and invasive species. We assessed the influence of LDT events on the invasive spread of raccoon rabies through Connecticut. We identified several putative LDT events, and developed a network-model to evaluate whether they became new foci for epidemic spread. LDT was fairly common, but many of the LDTs were isolated events that did not spread. Two putative LDT events did appear to become nascent foci that affected the epidemic in surrounding townships. In evaluating the role of LDT, we simultaneously revisited the problem of spatial heterogeneity. The spread of raccoon rabies is associated with forest cover—rabies moves up to three-times slower through the most heavily forested townships compared with those with less forestation. Forestation also modified the effect of rivers. In the best overall model, rabies did not cross the river separating townships that were heavily forested, and the spread slowed substantially between townships that were lightly forested. Our results suggest that spatial heterogeneity can be used to enhance the effects of rabies control by focusing vaccine bait distribution along rivers in lightly forested areas. LDT events are a concern, but this analysis suggests that at a local scale they can be isolated and managed.


title: Translating Research into Clinical Practice: Functional Recovery Post Total Hip Arthroplasty Using Outcomes Gathered in the Usual Physical Therapy Practice Setting

text: Abstract Purpose The purpose of this study is to determine the pattern of functional change after total hip arthroplasty (THA) in patients attending physical therapy (PT) in a usual care setting and to explore the effect of sex and time from surgery to the first physical therapy visit as potential prognostic factors influencing postoperative THA recovery. Study Design Adults with THA were retrospectively identified in an electronic medical record PT database (October 1, 2004-April 30, 2010). Hierarchical linear modeling was used to evaluate growth curves and individual variations in function using the Lower Extremity Function Scale (LEFS). Investigated predictors were: sex, age, start time, and PT visit. Results A total of 147 (81 female, 66 male) postoperative THA patients were included in the study; mean age was 62.7 years (SD 10.6, range 45-91 years). The majority (79%) of patients initiated PT <9 weeks postsurgery; predominately lower-functioning women started at ≥9 weeks. For patients initiating treatment at <9 weeks, the curvilinear slopes of recovery were similar between sexes, although the predicted levels of functional status were lower for females than for males (P = .041). Conclusions This study of usual physical therapy practice supports the findings from controlled studies that post-THA women enter and are discharged from outpatient PT with lower functional status than men. New findings suggest that functional status for early start patients steadily improves over 26 weeks postsurgery. Modeling change in clinical practice using outcomes measures acquired through usual practice can feasibly and adequately serve to guide decisions in the management of THA rehabilitation.


title: Comparative analysis of full genomic sequences among different genotypes of dengue virus type 3

text: BACKGROUND: Although the previous study demonstrated the envelope protein of dengue viruses is under purifying selection pressure, little is known about the genetic differences of full-length viral genomes of DENV-3. In our study, complete genomic sequencing of DENV-3 strains collected from different geographical locations and isolation years were determined and the sequence diversity as well as selection pressure sites in the DENV genome other than within the E gene were also analyzed. RESULTS: Using maximum likelihood and Bayesian approaches, our phylogenetic analysis revealed that the Taiwan's indigenous DENV-3 isolated from 1994 and 1998 dengue/DHF epidemics and one 1999 sporadic case were of the three different genotypes – I, II, and III, each associated with DENV-3 circulating in Indonesia, Thailand and Sri Lanka, respectively. Sequence diversity and selection pressure of different genomic regions among DENV-3 different genotypes was further examined to understand the global DENV-3 evolution. The highest nucleotide sequence diversity among the fully sequenced DENV-3 strains was found in the nonstructural protein 2A (mean ± SD: 5.84 ± 0.54) and envelope protein gene regions (mean ± SD: 5.04 ± 0.32). Further analysis found that positive selection pressure of DENV-3 may occur in the non-structural protein 1 gene region and the positive selection site was detected at position 178 of the NS1 gene. CONCLUSION: Our study confirmed that the envelope protein is under purifying selection pressure although it presented higher sequence diversity. The detection of positive selection pressure in the non-structural protein along genotype II indicated that DENV-3 originated from Southeast Asia needs to monitor the emergence of DENV strains with epidemic potential for better epidemic prevention and vaccine development.


title: E-SHOPPING changes and the state of E-grocery shopping in the U.S. evidence from national travel and time use surveys

text: Abstract In spite of the popularity of e-shopping, only 16% of US adults have ordered groceries online, and 7 out of 10 of those who currently buy groceries online do so at most twice a month. Understanding the determinants of e-grocery shopping is important for grocers, supply chain managers, and urban planners. In this context, we first explore how deliveries from online shopping have been changing over time. From our analysis of the 2009 and 2017 National Household Travel Surveys, we found that online shopping has been embraced by increasingly diverse households, although income, education, and some racial/ethnic differences persist. Our analysis of the 2017 American Time Use Survey shows that Americans are 24 times more likely to shop for groceries in stores than online. Moreover, in-store grocery shoppers are more likely to be female and unemployed, but less likely to belong to younger generations, to have less than a college degree, or to be African American. The gender imbalance in grocery shopping is larger online than in stores, but e-grocery shoppers do not otherwise differ from the general population. Future travel and e-shopping surveys (especially for e-grocery) should combine time use and travel questions with retrospective questions about online purchases.


title: Rapid Isothermal Amplification and Portable Detection System for SARS-CoV-2

text: The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs. As assay details and primer sequences become widely known, many laboratories could perform diagnostic tests using methods such as RT-PCR or isothermal RT-LAMP amplification. A key advantage of RT-LAMP based approaches compared to RT-PCR is that RT-LAMP is known to be robust in detecting targets from unprocessed samples. In addition, RT-LAMP assays are performed at a constant temperature enabling speed, simplicity, and point-of-use testing. Here, we provide the details of an RT-LAMP isothermal assay for the detection of SARS-CoV-2 virus with performance comparable to currently approved tests using RT-PCR. We characterize the assay by introducing swabs in virus spiked synthetic nasal fluids, moving the swab to viral transport medium (VTM), and using a volume of that VTM for performing the amplification without an RNA extraction kit. The assay has a Limit-of-Detection (LOD) of 50 RNA copies/μL in the VTM solution within 20 minutes, and LOD of 5000 RNA copies/μL in the nasal solution. Additionally, we show the utility of this assay for real-time point-of-use testing by demonstrating detection of SARS-CoV-2 virus in less than 40 minutes using an additively manufactured cartridge and a smartphone-based reader. Finally, we explore the speed and cost advantages by comparing the required resources and workflows with RT-PCR. This work could accelerate the development and availability of SARS-CoV-2 diagnostics by proving alternatives to conventional laboratory benchtop tests.


title: Cardiac and arrhythmic complications in patients with COVID‐19

text: In December 2019, the world started to face a new pandemic situation, the severe acute respiratory syndrome‐coronavirus 2 (SARS‐CoV‐2). Although coronavirus disease (COVID‐19) clinical manifestations are mainly respiratory, major cardiac complications are being reported. Cardiac manifestations etiology seems to be multifactorial, comprising direct viral myocardial damage, hypoxia, hypotension, enhanced inflammatory status, ACE2‐receptors downregulation, drug toxicity, endogenous catecholamine adrenergic status, among others. Studies evaluating patients with COVID‐19 presenting cardiac injury markers show that it is associated with poorer outcomes, and arrhythmic events are not uncommon. Besides, drugs currently used to treat the COVID‐19 are known to prolong the QT interval and can have a proarrhythmic propensity. This review focus on COVID‐19 cardiac and arrhythmic manifestations and, in parallel, makes an appraisal of other virus epidemics as SARS‐CoV, Middle East respiratory syndrome coronavirus, and H1N1 influenza.


title: Herbal plants and plant preparations as remedial approach for viral diseases

text: Herbal plants, plant preparations and phytoconstituents have proved useful in attenuating infectious conditions and were the only remedies available, till the advent of antibiotics (many being of plant origin themselves). Among infectious diseases, viral diseases in particular, remain the leading cause of death in humans globally. A variety of phytoconstituents derived from medicinal herbs have been extensively studied for antiviral activity. Based on this rationale, an online search was performed, which helped to identify a large number of plant species harboring antiviral molecules. These herbal sources have been reported individually or in combinations across a large number of citations studied. Activities against rabies virus, Human immunodeficiency virus, Chandipura virus, Japanese Encephalitis Virus, Enterovirus, Influenza A/H1N1 and other influenza viruses were discovered during the literature search. This review includes all such plant species exhibiting antiviral properties. The review also encompasses composition and methodologies of preparing various antiviral formulations around the globe. An elaborate section on the formulations filed for patent registration, along with non-patented formulations, has also been included in this article. To conclude, herbal sources provide researchers enormous scope to explore and bring out viable alternatives against viral diseases, considering non-availability of suitable drug candidates and increasing resistance to existing drug molecules for many emerging and re-emerging viral diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s13337-015-0276-6) contains supplementary material, which is available to authorized users.


title: Post-lockdown management of oncological priorities and postponed radiation therapy following the COVID-19 pandemic: Experience of the institut curie

text: • During COVID-19 lockdown, the number of radiotherapy treatments had to be reduced. • Given number of delayed treatments, the post-crisis period should be anticipated. • Data visualization software allowed to predict when peak activity would happen.


title: Overweight and obese men’s experiences in a sport-based weight loss intervention for men

text: In Western countries, such as Australia and the UK, a significantly greater proportion of men (relative to women) are overweight and obese, yet relatively few weight loss interventions have been developed that sufficiently target men. This lack of male-focused programming may be in part because ‘traditional’ weight loss programs are unappealing for what is considered a ‘hard-to-reach’ population. One program that appears to have such appeal for men is the MAN v FAT Football (MVFF) program, based out of the United Kingdom, which is designed for men with a body mass index of (or greater than) 27.5. MVFF encourages men’s participation in a community-based weight loss program that incentivizes weight loss through participation in a football league, and since 2016 MVFF has supported the weight loss efforts of several thousand men. Using MVFF as an exemplar, our aim was to derive insight into how men experience a male-only competitive, sport-based weight loss program. We recruited twenty-seven players (Mage = 41.13, SD = 9.93), and ten coaches (Mage = 31.8, SD = 11.55) from program locations throughout the United Kingdom. Using semi-structured interviews and thematic analysis, we identified several appraisal aspects of the program that players and coaches considered important, including the appeal of sport, competition on a level playing field, being part of a team, camaraderie, accountability, men sharing issues with other men, gender-sensitized environment, likeminded and similar men, and perceptions that traditional weight loss programs are tailored towards women. Player experiences (i.e., competence and enjoyment) and functional supports in the program (e.g., player handbook, weight loss coach) were reported to drive outcomes of effective weight loss and program retention. Interventions aiming to target men may be more successful working with rather than against formulations of identity such as masculinities, and this can be achieved by tailoring program content (e.g., messaging), settings (e.g., among men sharing similar characteristics such as body-type or goals), and mode of delivery (e.g., through organized sports, and leveraging competition to drive healthy behaviours).


title: Host-Species Transferrin Receptor 1 Orthologs Are Cellular Receptors for Nonpathogenic New World Clade B Arenaviruses

text: The ability of a New World (NW) clade B arenavirus to enter cells using human transferrin receptor 1 (TfR1) strictly correlates with its ability to cause hemorrhagic fever. Amapari (AMAV) and Tacaribe (TCRV), two nonpathogenic NW clade B arenaviruses that do not use human TfR1, are closely related to the NW arenaviruses that cause hemorrhagic fevers. Here we show that pseudotyped viruses bearing the surface glycoprotein (GP) of AMAV or TCRV can infect cells using the TfR1 orthologs of several mammalian species, including those of their respective natural hosts, the small rodent Neacomys spinosus and the fruit bat Artibeus jamaicensis. Mutation of one residue in human TfR1 makes it a functional receptor for TCRV, and mutation of four residues makes it a functional receptor for AMAV. Our data support an in vivo role for TfR1 in the replication of most, if not all, NW clade B arenaviruses, and suggest that with modest changes in their GPs the nonpathogenic arenaviruses could use human TfR1 and emerge as human pathogens.


title: Toona sinensis Inhibits Murine Leukemia WEHI-3 Cells and Promotes Immune Response In Vivo

text: Toona sinensis (TS) is one of the most popular vegetarian dishes in Taiwan. It has been shown to exhibit antioxidant, antiangiogenic, antiatherosclerotic, and anticancer properties. In this study, we demonstrated the ability of aqueous leaf extracts from TS to promote immune responses in BALB/c mice and to exhibit anti-leukemia activity in murine WEHI-3 cells. BALB/c mice were injected intravenously with WEHI-3 cells and then treated orally with TS (50 mg/kg). In vivo study showed that TS treatment reduced liver and spleen enlargement in WEHI-3 bearing mice compared with the untreated group. Furthermore, TS also decreased white blood cells (WBC), indicating inhibition of differentiation of the precursor of macrophages in WEHI-3 bearing mice. Treatment of WEHI-3 cells with TS (0-75 μg/mL for 24 hours) significantly reduced cell viability. Furthermore, TS treatment–induced late apoptosis was confirmed by Annexin-V/PI staining. Western blot analyses revealed that treatment of WEHI-3 cells with TS statistically increased the protein expression level of cytochrome c in the cytoplasm and activates caspase-3. Notably, TS treatment caused a dramatic reduction in Bcl-2 and increase in Bax protein levels. TS may disturb the Bcl-2 and Bax protein ratio and induce apoptosis. This reports confirms the antitumor activity of this nutritious vegetable potentially against leukemia.


title: Proficiency testing for the detection of Middle East respiratory syndrome coronavirus demonstrates global capacity to detect Middle East respiratory syndrome coronavirus

text: The first reported case of Middle East respiratory syndrome coronavirus (MERS‐CoV) infection was identified in Saudi Arabia in September 2012, since which time there have been over 2000 laboratory‐confirmed cases, including 750 deaths in 27 countries. Nucleic acid testing (NAT) is the preferred method for the detection of MERS‐CoV. A single round of a Proficiency Testing Program (PTP) was used to assess the capability of laboratories globally to accurately detect the presence of MERS‐CoV using NAT. A panel of eleven lyophilized specimens containing different viral loads of MERS‐CoV, common coronaviruses, and in vitro RNA transcripts was distributed to laboratories in all six World Health Organization regions. A total of 96 laboratories from 79 countries participating in the PTP, with 76 of 96 (79.2%) reporting correct MERS‐CoV results for all nine scored specimens. A further 10 laboratories (10.4%) scored correctly in eight of nine specimens of the PTP. The majority of laboratories demonstrated satisfactory performance in detecting the presence of MERS‐CoV using NAT. However, some laboratories require improved assay sensitivity, reduced cross contamination of samples, and improved speciation of coronavirus subtypes for potentially complex clinical specimens. Further PTP and enhanced links with expert laboratories globally may improve the laboratory performance.


title: Understanding of guidance for acupuncture and moxibustion interventions on COVID-19 (Second edition) issued by CAAM

text: Abstract At present, the situation of global fight against COVID-19 is serious. WHO (World Health Organization)-China Joint Mission fully confirms the success of “China's model” against COVID-19 in the report. In fact, one particular power in “China's model” is acupuncture and moxibustion of traditional Chinese medicine. To better apply “non-pharmaceutic measures”—the external technique of traditional Chinese medicine, in the article, the main content of Guidance for acupuncture and moxibustion interventions on COVID-19 (Second edition) issued by China Association of Acupuncture–Moxibution is introduced and the discussion is stressed on the selection of moxibustion device and the duration of its exertion.


title: Prevalence and impact of acute renal impairment on COVID-19: a systematic review and meta-analysis

text: BACKGROUND: The aim of this study is to assess the prevalence of abnormal urine analysis and kidney dysfunction in COVID-19 patients and to determine the association of acute kidney injury (AKI) with the severity and prognosis of COVID-19 patients. METHODS: The electronic database of Embase and PubMed were searched for relevant studies. A meta-analysis of eligible studies that reported the prevalence of abnormal urine analysis and kidney dysfunction in COVID-19 was performed. The incidences of AKI were compared between severe versus non-severe patients and survivors versus non-survivors. RESULTS: A total of 24 studies involving 4963 confirmed COVID-19 patients were included. The proportions of patients with elevation of sCr and BUN levels were 9.6% (95% CI 5.7–13.5%) and 13.7% (95% CI 5.5–21.9%), respectively. Of all patients, 57.2% (95% CI 40.6–73.8%) had proteinuria, 38.8% (95% CI 26.3–51.3%) had proteinuria +, and 10.6% (95% CI 7.9–13.3%) had proteinuria ++ or +++. The overall incidence of AKI in all COVID-19 patients was 4.5% (95% CI 3.0–6.0%), while the incidence of AKI was 1.3% (95% CI 0.2–2.4%), 2.8% (95% CI 1.4–4.2%), and 36.4% (95% CI 14.6–58.3%) in mild or moderate cases, severe cases, and critical cases, respectively. Meanwhile, the incidence of AKI was 52.9%(95% CI 34.5–71.4%), 0.7% (95% CI − 0.3–1.8%) in non-survivors and survivors, respectively. Continuous renal replacement therapy (CRRT) was required in 5.6% (95% CI 2.6–8.6%) severe patients, 0.1% (95% CI − 0.1–0.2%) non-severe patients and 15.6% (95% CI 10.8–20.5%) non-survivors and 0.4% (95% CI − 0.2–1.0%) survivors, respectively. CONCLUSION: The incidence of abnormal urine analysis and kidney dysfunction in COVID-19 was high and AKI is closely associated with the severity and prognosis of COVID-19 patients. Therefore, it is important to increase awareness of kidney dysfunction in COVID-19 patients.


docid: hleylymf

title: Co-expressing GP5 and M proteins under different promoters in recombinant modified vaccinia virus ankara (rMVA)-based vaccine vector enhanced the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV)

text: The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP5, M, and N. Protein GP5 and M have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. There were some attempts on expressing GP5 or M in DNA vaccine and adenovirus to arouse humoral and cellular immune responses, but few papers have been reported on that the immune response can be difference because of the expression patterns of GP5 and M proteins in the recombinant virus. In this article, four recombinant viruses that expressed GP5 and M proteins of PRRSV in the modified vaccinia virus ankara (MVA) with different expression patterns were made. In these recombinant virus (rMVAs), GP5 and M proteins were expressed in MVA in the same virus but under the control of two promoters (rMVA-GP5/M), or as a fusion protein under one promoter (rMVA-GP5-M), or separately (rMVA-GP5 and rMVA-M). The humoral and cellular immune responses for the four recombinant viruses were evaluated with mouse model. Every mouse was inoculated with 5 × 10(5) TCID(50) of the different rMVAs and boosted 3 weeks later. Neutralizing antibody titers for each group were detected with virus neutralization test assay weekly after the primary inoculation for 13 weeks to evaluate the humoral immune response. The production of gamma interferon (IFN-γ), interleukin-2 (IL-2), and interleukin-4 (IL-4) was detected in splenocytes of rMVA-inoculated mice at 30, 60, and 90 days post inoculation to evaluate the cellular immune response. Results showed that rMVA-GP5 and rMVA-M cannot induce obvious humoral and cellular immune responses; rMVA-GP5-M inoculated group developed better immune responses than rMVA-GP5 and rMVA-M inoculated groups; however, mice inoculated with rMVA-GP5/M maintained the strongest cellular response against PRRS and consistently enhanced the anti-PRRSV humoral responses. The strategy of co-expressing PRRSV GP5 and M protein in MVA under the control of different promoters might be an attractive method for future PRRSV vaccine design.


title: Oligonucleotide aptamers: promising and powerful diagnostic and therapeutic tools for infectious diseases

text: Summary The entire human population is at risk of infectious diseases worldwide. Thus far, the diagnosis and treatment of human infectious diseases at the molecular and nanoscale levels have been extremely challenging tasks because of the lack of effective probes to identify and recognize biomarkers of pathogens. Oligonucleotide aptamers are a class of small nucleic acid ligands that are composed of single-stranded DNA (ssDNA) or RNA and act as affinity probes or molecular recognition elements for a variety of targets. These aptamers have an exciting potential for diagnose and/or treatment of specific diseases. In this review, we highlight areas where aptamers have been developed as diagnostic and therapeutic agents for both bacterial and viral infectious diseases as well as aptamer-based detection.