title: Isolation and recombinant analysis of variants of porcine epidemic diarrhea virus strains from Beijing, China

text: Porcine epidemic diarrhea virus (PEDV) is a highly infectious virus infecting pigs with high morbidity, especially for newborn piglets. Several PEDV strains were isolated from the intestinal tracts of diarrheic piglets from the Beijing area, China. Sequencing of the whole-genome of the PEDV isolates (GenBank numbers MG546687-MG546690) yielded sequences of 28033–28038 nt. The phylogenetic tree revealed that these strains from the Beijing area belonged to group II, while the vaccine strain, CV777, belonged to group I. We also determined the genetic correlation between these strains and CV777 strain. However, it showed that these strains in the Beijing area had unique mutations. The sequence identity of PEDV strains showed that these strains are most similar to these strains LZW, CH/JX-1/2013, USAIllinois972013, USAKansas1252014, CH/GDZQ/2014, SHQPYM2013, AJ1102, CHZMDZY11, KoreaK14JB01, and CHYJ130330, respectively. The possible recombination events indicate that PEDV in this studies were possibly recombinant strain formed by parent strains USAIllinois972013, KoreaK14JB01, CHYJ130330, and CHZMDZY11. These PEDV strains has been genetic recombination and mutations. The variant strains characterized in this study help to the evolutionary analysis of PEDV.


title: 6. Asthma: Factors underlying inception, exacerbation, and disease progression

text: Asthma is a heterogeneous disorder that is characterized by variable airflow obstruction, airway inflammation and hyperresponsiveness, and reversibility either spontaneously or as a result of treatment. Multiple causes no doubt exist for both its inception and symptom exacerbation once the disease is established. Factors underlying inception can range from viral respiratory tract infections in infancy to occupational exposures in adults. Factors underlying asthma exacerbations include allergen exposure in sensitized individuals, viral infections, exercise, irritants, and ingestion of nonsteroidal anti-inflammatory agents among others. Exacerbating factors might include one or all of these exposures and vary both among and within patients. Asthma treatment is determined to a large extent after an assessment of severity, which can be variable over time and assessed in 2 domains: impairment (current) and risk (long-term consequences). Unfortunately, despite the availability of effective therapies, suboptimal asthma control exists in many patients on a worldwide basis. The future development of novel therapies and treatment paradigms should address these disparities.


title: Transmission genetics of drug-resistant hepatitis C virus

text: Antiviral development is plagued by drug resistance and genetic barriers to resistance are needed. For HIV and hepatitis C virus (HCV), combination therapy has proved life-saving. The targets of direct-acting antivirals for HCV infection are NS3/4A protease, NS5A phosphoprotein and NS5B polymerase. Differential visualization of drug-resistant and -susceptible RNA genomes within cells revealed that resistant variants of NS3/4A protease and NS5A phosphoprotein are cis-dominant, ensuring their direct selection from complex environments. Confocal microscopy revealed that RNA replication complexes are genome-specific, rationalizing the non-interaction of wild-type and variant products. No HCV antivirals yet display the dominance of drug susceptibility shown for capsid proteins of other viruses. However, effective inhibitors of HCV polymerase exact such high fitness costs for drug resistance that stable genome selection is not observed. Barriers to drug resistance vary with target biochemistry and detailed analysis of these barriers should lead to the use of fewer drugs.


title: Pattern of liver injury in adult patients with COVID-19: a retrospective analysis of 105 patients

text: BACKGROUND: Recent studies reported that patients with coronavirus disease-2019 (COVID-19) might have liver injury. However, few data on the combined analysis and change patterns of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil) have been shown. METHODS: This is a single-center retrospective study. A total of 105 adult patients hospitalized for confirmed COVID-19 in Beijing Ditan Hospital between January 12, and March 17, 2020 were included, and divided into mild group (n = 79) and severe group(n = 26). We compared liver functional test results between the two groups. Category of ALT change during the disease course was also examined. RESULTS: 56.2% (59/105) of the patients had unnormal ALT, AST, or total TBil throughout the course of the disease, but in 91.4% (96/105) cases the level of ALT, AST or TBil ≤3 fold of the upper limit of normal reference range (ULN). The overall distribution of ALT, AST, and TBil were all significantly difference between mild and severe group (P < 0.05). The percentage of the patients with elevated both ALT and AST was 12.7% (10/79) in mild cases vs. 46.2% (12/26) in severe cases (P = 0.001). 34.6% (9/26) severe group patients started to have abnormal ALT after admission, and 73.3% (77/105) of all patients had normal ALT before discharge. CONCLUSIONS: Elevated liver function index is very common in patients with COVID-19 infection, and the level were less than 3 × ULN, but most are reversible. The abnormality of 2 or more indexes is low in the patients with COVID-19, but it is more likely to occur in the severe group.


title: Neutral metalloaminopeptidases APN and MetAP2 as newly discovered anticancer molecular targets of actinomycin D and its simple analogs

text: The potent transcription inhibitor Actinomycin D is used with several cancers. Here, we report the discovery that this naturally occurring antibiotic inhibits two human neutral aminopeptidases, the cell-surface alanine aminopeptidase and intracellular methionine aminopeptidase type 2. These metallo-containing exopeptidases participate in tumor cell expansion and motility and are targets for anticancer therapies. We show that the peptide portions of Actinomycin D and Actinomycin X(2) are not required for effective inhibition, but the loss of these regions changes the mechanism of interaction. Two structurally less complex Actinomycin D analogs containing the phenoxazone chromophores, Questiomycin A and Actinocin, appear to be competitive inhibitors of both aminopeptidases, with potencies similar to the non-competitive macrocyclic parent compound (K(i) in the micromolar range). The mode of action for all four compounds and both enzymes was demonstrated by molecular modeling and docking in the corresponding active sites. This knowledge gives new perspectives to Actinomycin D's action on tumors and suggests new avenues and molecules for medical applications.


title: Absolute quantitation of infectious salmon anaemia virus using different real-time reverse transcription PCR chemistries

text: Abstract Routine laboratory diagnosis of infectious salmon anaemia virus (ISAV) infection is primarily by reverse transcription polymerase chain reaction (RT-PCR) because of the high sensitivity and rapid turnaround time of the test. This paper describes methods for highly reproducible absolute viral load measurements using external standard curves generated with either ISAV recombinant plasmid DNA (pDNA) standards or transcribed RNA standards prepared by in vitro transcription with T7 RNA polymerase, and using a two tube real-time or quantitative (q)RT-PCR with SYBR® Green I chemistry and a single tube qRT-PCR with TaqMan® probe chemistry. When applied to virus samples of known virus titer for the highly pathogenic ISAV strain NBISA01 and the low pathogenic ISAV strain RPC/NB-04-085-1, both methods showed a 100-fold lower detectable titer for RPC/NB-04-085-1 but with a higher number of viral RNA molecules compared to NBISA01. Overall, the SYBR® Green I method overestimated copy numbers in samples having equivalent Ct values with the TaqMan® probe method. Taken together, the findings suggest that the TaqMan® probe method with the in vitro transcribed RNA standard curve is the preferred method for reliable and rapid quantitation of ISAV in samples.


title: A case–control study of pathogen and lifestyle risk factors for diarrhoea in dogs

text: Abstract Diarrhoea is a common and multi-factorial condition in dogs, the aetiology of which is often incompletely understood. A case–control study was carried out to compare the carriage of some common canine enteric pathogens (enteric coronavirus, parvovirus, distemper, endoparasites, Campylobacter and Salmonella spp.), as well as lifestyle factors such as vaccination history, diet and contact with other species, in dogs presenting at first opinion veterinary practices with and without diarrhoea. Multivariable conditional logistic regression showed that dogs in the study which scavenged or had had a recent change of diet (OR 3.5, p =0.002), had recently stayed in kennels (OR 9.5, p =0.01), or were fed a home-cooked diet (OR 4, p =0.002) were at a significantly greater risk of diarrhoea, whilst being female (OR 0.4, p =0.01), currently up to date with routine vaccinations (OR 0.4, p =0.05) and having contact with horse faeces (OR 0.4, p =0.06) were associated with a reduced risk. None of the pathogens tested for was a significant factor in the final multivariable model suggesting that in this predominantly vaccinated population, diarrhoea may be more associated with lifestyle risk factors than specific pathogens.


title: ACAM2000 clonal Vero cell culture vaccinia virus (New York City Board of Health strain) – a second-generation smallpox vaccine for biological defense

text: Summary The threat of smallpox as a biological weapon has spurred efforts to create stockpiles of vaccine for emergency preparedness. In lieu of preparing vaccine in animal skin (the original method), we cloned vaccinia virus (New York City Board of Health strain, Dryvax®) by plaque purification and amplified the clone in cell culture. The overarching goal was to produce a modern vaccine that was equivalent to the currently licensed Dryvax® in its preclinical and clinical properties, and could thus reliably protect humans against smallpox. A variety of clones were evaluated, and many were unacceptably virulent in animal models. One clonal virus (ACAM1000) was selected and produced at clinical grade in MRC-5 human diploid cells. ACAM1000 was comparable to Dryvax® in immunogenicity and protective activity but was less neurovirulent for mice and nonhuman primates. To meet requirements for large quantities of vaccine after the events of September 11th 2001, the ACAM1000 master virus seed was used to prepare vaccine (designated ACAM2000) at large scale in Vero cells under serum-free conditions. The genomes of ACAM1000 and ACAM2000 had identical nucleotide sequences, and the vaccines had comparable biological phenotypes. ACAM1000 and ACAM2000 were evaluated in three Phase 1 clinical trials. The vaccines produced major cutaneous reactions and evoked neutralizing antibody and cell-mediated immune responses in the vast majority of subjects and had a reactogenicity profile similar to that of Dryvax®.


title: Simulated gastrointestinal tract metabolism and pharmacological activities of water extract of Scutellaria baicalensis roots

text: Abstract Ethnopharmacological relevance . Scutellaria baicalensis Georgi (Labiatae) is a well-known traditional Chinese medicine to treat inflammation, cardiovascular diseases, respiratory and gastrointestinal infections, etc. The present study was to understand the metabolism of the root of Scutellaria baicalensis (a.k.a. Huangqin in Chinese) in the gastrointestinal tract and the correlation between the metabolites and their respective pharmacological activities. Materials and methods The water extract of the root of Scutellaria baicalensis (WESB) was incubated with simulated gastric and intestinal juices, and human fecal microflora for 24h at 37°C. The HPLC–DAD analysis was used to monitor the in vitro metabolic process and identify its metabolites by comparing their absorption spectrum and retention time with those of chemical references. The in vitro anticomplementary and antimicrobial activity was evaluated with hemolysis assay, agar disc-diffusion method and MIC value, respectively. Results Main constituents of WESB remain unchanged during the incubation with simulated gastric juice (pH=1.5) and intestinal juice (pH=6.8), whereas four flavones, baicalin, wogonoside, oroxyloside and norwogonoside were metabolized into their respective aglycons by human intestinal bacteria. All four metabolites were demonstrated to have higher anticomplementary and antimicrobial activity than those of WESB. The anticomplementary active metabolites were identified to be baicalein, oroxylin A and norwogonin, among them, norwogonin is the most active compound. Conclusion The presence of intestinal bacteria is demonstrated to play an important role in the gastrointestinal metabolism of WESB, and the pharmacological effects of Scutellaria baicalensis may be dependent on the intestinal bacteria metabolism.


Reliability and usefulness of a rapid IgM‐IgG antibody test for the diagnosis of SARS-CoV-2 infection: a preliminary report.


title: Availability of feces-free areas in rodent shoebox cages

text: The Guide for the Care and Use of Laboratory Animals (the Guide) recommends that terrestrial mammals be provided space free of urine and feces in which to rest. To evaluate the feasibility of meeting this recommendation, the author examined the availability of feces-free resting areas in standard rodent cages over time. Adult rodents (C57BL/6J mice and Wistar rats) were housed singly, in pairs or in trios in shoebox cages at densities that met the space recommendations of the Guide. As housing density increased, the availability of unsoiled resting space declined. For C57BL/6J mice housed singly, in pairs or in trios, most cages lacked unsoiled resting area within 3–6 days (depending on cage size), 2 days or 1 day, respectively. Similarly, for Wistar rats housed singly, in pairs or in trios, most cages lacked adequate unsoiled resting space within 3 days, 2 days or 1 day, respectively. Because most cages lacked adequate unsoiled resting space within 3 days of housing animals, the author concludes that standard cage change frequencies of once a week for adult C57BL/6J mice and twice a week for adult Wistar rats may be inadequate to provide unsoiled resting areas for rodents.


title: Co-infections, genetic, and antigenic relatedness of avian influenza H5N8 and H5N1 viruses in domestic and wild birds in Egypt

text: A total of 50 poultry farms of commercial broilers (N = 39) and commercial layers (N = 11) suffered from respiratory problems and mortality during the period from January 2016 to December 2017 were investigated. Also, samples were collected from quail (N = 4), Bluebird (Sialis, N = 1), and Greenfinch (Chloris chloris, N = 1) for analysis. Respiratory viral pathogens were screened by PCR and positive samples were subjected to virus isolation and genetic identification. Antigenic relatedness of isolated avian influenza (AI) H5 subtype was evaluated using cross-hemagglutination inhibition. Results revealed that the incidence of single virus infections in commercial broilers was 64.1% (25/39), with the highest incidence for ND (33.3%) and H9N2 (20.5%), followed by H5N1 (7.7%) and H5N8 (2.7). Meanwhile, H9N2/ND mixed infection was the most observed case (7.7%). Other mixed infections H5N1/ND, H5N1/H9N2/ND, H5N1/H9N2/ND/IB, H9N2/IB, and H9N2/ILT were also observed (2.6% each). In commercial layers, H5N1 and ILT were the only detected single infections (18.1% each). Mixed H9N2/ND was the most predominant infection in layers (27.3%). Other mixed infections of H9N2/IB, H5N1/H5N8/H9N2, and H9N2/ND/IB were observed in 3 separate farms (9.1% each). The H5N8 virus was detected in one quail farm and 2 out of 3 wild bird's samples. Partial HA gene sequence analysis showed the clustering of the selected AI H5N8 within the 2.3.4.4 clade, while H5N1 clustered with the clade 2.2.1.2. Interestingly, the H5N8 isolated from chickens possessed 6 amino acids substitutions at HA1 compared to those isolated from wild birds with low antigenic relatedness to AI H5N1 clades 2.2.1 or 2.2.1.2. In conclusion, mixed viral infections were observed in both broiler and layer chickens in Egypt. The detected triple H5N1, H9N2, and H5N8 influenza co-infection raises the concern of potential AI epidemic strain emergence. The low genetic and antigenic relatedness between AI H5N1 and H5N8 viruses suggest the need for modification of vaccination strategies of avian influenza in Egypt along with strict biosecurity measures.


Teledermatology for patient management, dermatology education and research during the COVID‐19 pandemic


title: Serum KL‐6 concentrations as a novel biomarker of severe COVID‐19

text: Severe acute respiratory syndrome coronavirus 2–induced direct cytopathic effects against type I and II pneumocytes mediate lung damage. Krebs von den Lungen‐6 (KL‐6) is mainly produced by damaged or regenerating alveolar type II pneumocytes. This preliminary study analyzed serum concentrations of KL‐6 in patients with coronavirus disease (COVID‐19) to verify its potential as a prognostic biomarker of severity. Twenty‐two patients (median age [interquartile range] 63 [59‐68] years, 16 males) with COVID‐19 were enrolled prospectively. Patients were divided into mild‐moderate and severe groups, according to respiratory impairment and clinical management. KL‐6 serum concentrations and lymphocyte subset were obtained. Peripheral natural killer (NK) cells/µL were significantly higher in nonsevere patients than in the severe group (P = .0449) and the best cut‐off value was 119 cells/µL. KL‐6 serum concentrations were significantly higher in severe patients than the nonsevere group (P = .0118). Receiver operating characteristic analysis distinguished severe and nonsevere patients according to KL‐6 serum levels and the best cut‐off value was 406.5 U/mL. NK cell analysis and assay of KL‐6 in serum can help identify severe COVID‐19 patients. Increased KL‐6 serum concentrations were observed in patients with severe pulmonary involvement, revealing a prognostic value and supporting the potential usefulness of KL‐6 measurement to evaluate COVID‐19 patients' prognosis.


title: Two-amino acids change in the nsp4 of SARS coronavirus abolishes viral replication

text: Abstract Infection with coronavirus rearranges the host cell membrane to assemble a replication/transcription complex in which replication of the viral genome and transcription of viral mRNA occur. Although coexistence of nsp3 and nsp4 is known to cause membrane rearrangement, the mechanisms underlying the interaction of these two proteins remain unclear. We demonstrated that binding of nsp4 with nsp3 is essential for membrane rearrangement and identified amino acid residues in nsp4 responsible for the interaction with nsp3. In addition, we revealed that the nsp3-nsp4 interaction is not sufficient to induce membrane rearrangement, suggesting the participation of other factors such as host proteins. Finally, we showed that loss of the nsp3-nsp4 interaction eliminated viral replication by using an infectious cDNA clone and replicon system of SARS-CoV. These findings provide clues to the mechanism of the replication/transcription complex assembly of SARS-CoV and could reveal an antiviral target for the treatment of betacoronavirus infection.


title: Structural basis for catalysis and ubiquitin recognition by the Severe acute respiratory syndrome coronavirus papain‐like protease

text: Papain‐like protease (PL(pro)) is one of two cysteine proteases involved in the proteolytic processing of the polyproteins of Severe acute respiratory syndrome coronavirus (SARS‐CoV). PL(pro) also shows significant in vitro deubiquitinating and de‐ISGylating activities, although the detailed mechanism is still unclear. Here, the crystal structure of SARS‐CoV PL(pro) C112S mutant in complex with ubiquitin (Ub) is reported at 1.4 Å resolution. The Ub core makes mostly hydrophilic interactions with PL(pro), while the Leu‐Arg‐Gly‐Gly C‐terminus of Ub is located in the catalytic cleft of PL(pro), mimicking the P4–P1 residues and providing the first atomic insights into its catalysis. One of the O atoms of the C‐terminal Gly residue of Ub is located in the oxyanion hole consisting of the main‐chain amides of residues 112 and 113. Mutations of residues in the PL(pro)–Ub interface lead to reduced catalytic activity, confirming their importance for Ub binding and/or catalysis. The structure also revealed an N‐cyclohexyl‐2‐aminethanesulfonic acid molecule near the catalytic triad, and kinetic studies suggest that this binding site is also used by other PL(pro) inhibitors. Overall, the structure provides a foundation for understanding the molecular basis of coronaviral PL(pro) catalysis.


title: Viral Findings in Adult Hematological Patients with Neutropenia

text: BACKGROUND: Until recently, viral infections in patients with hematological malignancies were concerns primarily in allogeneic hematopoietic stem cell transplant (HSCT) recipients. During the last years, changed treatment regimens for non-transplanted patients with hematological malignancies have had potential to increase the incidence of viral infections in this group. In this study, we have prospectively investigated the prevalence of a broad range of respiratory viruses in nasopharyngeal aspirate (NPA) as well as viruses that commonly reactivate after allogeneic HSCT. METHODOLOGY/PRINCIPAL FINDINGS: Patients with hematological malignancies and therapy induced neutropenia (n = 159) were screened regarding a broad range of common respiratory viruses in the nasopharynx and for viruses commonly detected in severely immunosuppressed patients in peripheral blood. Quantitative PCR was used for detection of viruses. A viral pathogen was detected in 35% of the patients. The detection rate was rather similar in blood (22%) and NPA (18%) with polyoma BK virus and rhinovirus as dominating pathogens in blood and NPA, respectively. Patients with chronic lymphocytic leukemia (CLL) (p<0.01) and patients with fever (p<0.001) were overrepresented in the virus-positive group. Furthermore, viral findings in NPA were associated with upper respiratory symptoms (URTS) (p<0.0001). CONCLUSIONS/SIGNIFICANCE: Both respiratory viral infections and low titers of viruses in blood from patients with neutropenia were common. Patients with CLL and patients with fever were independently associated to these infections, and viral findings in NPA were associated to URTS indicating active infection. These findings motivate further studies on viruses' impact on this patient category and their potential role as causative agents of fever during neutropenia.


title: Signature patterns revealed by microarray analyses of mice infected with influenza virus A and Streptococcus pneumoniae

text: Abstract We used cDNA microarrays to identify differentially expressed genes in mice in response to infections with influenza virus A/PR/8/34 (H1N1) and Streptococcus pneumoniae. Expression microarray analysis showed up-regulation and down-regulation of many genes involved in the defense, inflammatory response and intracellular signaling pathways including chemokine, apoptosis, MAPK, Notch, Jak-STAT, T-cell receptor and complement and coagulation cascades. We have revealed signature patterns of gene expression in mice infected with two different classes of pathogens: influenza virus A and S. pneumoniae. Quantitative real-time RT-PCR results confirmed microarray results for most of the genes tested. These studies document clear differences in gene expression profiles between mice infected with influenza virus A and S. pneumoniae. Identification of genes that are differentially expressed after respiratory infections can provide insights into the mechanisms by which the host interacts with different pathogens, useful information about stage of diseases and selection of suitable targets for early diagnosis and treatments. The advantage of this novel approach is that the detection of pathogens is based on the differences in host gene expression profiles in response to different pathogens instead of detecting pathogens directly.


title: A Continuous Renal Replacement Therapy Protocol for Patients with Acute Kidney Injury in Intensive Care Unit with COVID-19

text: COVID-19 often leads to acute respiratory distress syndrome complicated by acute kidney injury (AKI). The indications for renal replacement therapy for these patients are those commonly accepted to treat AKI. We describe a continuous veno-venous haemodialysis (CVVHD) protocol for AKI, which aims to provide the best treatment according to the particular patient’s and medical personnels’ needs in biohazard settings with limited human and technological resources. We designed a CVVHD protocol with a high cut-off (HCO) filter in regional citrate anticoagulation (RCA). The HCO filter in diffusion determines the enhanced cytokines clearance with less filter clotting due to a lower filtration fraction. In our hospital, at the beginning of the pandemic outbreak, we treated seven COVID-19 patients with AKI stage 2 and 3 and recorded the circuit lifespan and the number of interventions on monitors. CVVHD in RCA appears to be safe, effective and easy to be performed in a biohazard scenario using lower blood flows and less bag changes with fluid savings, a biohazard reduction and sparing of resources. Although the data come from a very small cohort, our protocol seems related to a low mortality.


title: Respiratory virus is a real pathogen in immunocompetent community-acquired pneumonia: comparing to influenza like illness and volunteer controls

text: BACKGROUND: Viral pathogens were more commonly reported than previously estimated in community-acquired pneumonia (CAP) patients. However, the real role of virus was still controversial. METHODS: Consecutive adult patients with CAP between April and December, 2009 were prospectively enrolled. A four-fold or greater increase of IgG-titres against respiratory viruses in pair sera was tested by means of hemagglutination inhibition assay or indirect immunofluorescence. Swab samples were tested by cell culture and/or nucleic amplification tests. Viral etiology was considered definitive if at least one of the above tests was positive. RESULTS: Viral etiology was established in fifty-two (34.9%) of 149 CAP patients, twenty-two (81.5%) of 27 influenza like illness patients, and none of 75 volunteer controls. Forty-seven CAP patients were infected by a single virus (24 influenza A virus, 5 influenza B, 10 parainfluenza virus type 3 [PIV-3], 2 PIV-1, 2 adenovirus, 2 human rhinovirus and 2 coronavirus OC43), five cases by two or three viruses co-infection. Fever ≥ 39°C (66.7%), fatigue (64.6%), and purulent sputum (52.1%) was the most common symptoms in viral pneumonia patients. On multivariate analysis, myalgia was included in the model for pneumonia associated with influenza infection. In the CURB-65 model only influenza infection was found independently associated with severe disease (CURB-65 score ≥ 3) out of variables, including age(years), sex, current smoking status, sick contact with febrile patients, numbers of comorbidity, presence of influenza infection, presence of PIV infection, with P = 0.021, OR 7.86 (95% CI 1.37-45.04). CONCLUSION: Respiratory virus was not a bystander, but pathogenic in pneumonia and was a common cause of CAP.


Abstract 34. Inoculation of schizophrenic brains to rodents and primates: behavioral and neuropathological observations


title: Experimental inoculation of adult dairy cows with bovine coronavirus and detection of coronavirus in feces by RT-PCR

text: A reverse transcriptase PCR (RT-PCR) targeting a 407 bp fragment of the nucleocapsid gene of bovine coronavirus (BCV) was developed for detection of BCV RNA in feces of experimentally inoculated cattle. The sensitivity and specificity of the RT-PCR were confirmed using tissue culture-adapted BCV strains and feces of 2 calves inoculated with BCV. Ten nonpregant, BCV seropositive, adult dairy cows were inoculated with winter dysentery (WD) (n = 8) or calf diarrhea (CD) (n = 2) strains of BCV intranasally and orally (n = 2) or through a surgically-placed duodenal catheter (n = 8) with and without dexamethasone treatment or feeding ice water. The 6 cows inoculated with BCV intranasally and through a duodenal catheter (2 of 2 cows given CD BCV and 4 of 6 cows given WD BCV) developed mild diarrhea, and BCV was detected in diarrheal feces by RT-PCR, ELISA or immune electron microscopy. These results suggest that CD and WD strains of BCV can cause diarrhea in adult cows in conjunction with host or environmental factors and that RT-PCR might be useful to diagnose BCV infections in calves and adult cows.


New Insights on the Structure and Morphogenesis of Berne Virus


title: Temporal changes in cytokine/chemokine profiles and pulmonary involvement in severe acute respiratory syndrome

text: Objective and background: Pathological changes in severe acute respiratory syndrome (SARS) suggest that SARS sequelae are associated with dysregulation of cytokine and chemokine production. To improve understanding of the immuno‐pathological processes involved in lung injury associated with SARS, the temporal changes in cytokine/chemokine profiles in the sera of SARS patients were compared with those of patients with community‐acquired pneumonia (CAP), according to the degree of lung involvement. Methods: Serum levels of 11 cytokines and chemokines, in 14 patients with SARS and 24 patients with CAP, were serially checked using a bead‐based multiassay system. Sera from 12 healthy subjects were used as normal controls. Results: The serum levels of interferon‐γ‐inducible protein‐10 (IP‐10), IL‐2 and IL‐6 were significantly elevated during SARS infection. In patients with CAP, but not in those with SARS, the levels of interferon‐γ, IL‐10, IL‐8 and monokine induced by interferon‐γ (MIG) were significantly elevated compared with the levels in healthy controls. Among the chemokines/cytokines, IL‐6 levels correlated most strongly with radiographic scores (r = 0.62). The elevation of IP‐10 and IL‐2 antedated the development of chest involvement and reached peak levels earlier than the radiographic scores. In contrast, the dynamic changes in IL‐6, C‐reactive protein and neutrophils occurred synchronously with the changes in radiographic scores. The mean ratio of IL‐6 to IL‐10 in SARS patients (4.84; range 0.41–21) was significantly higher than that in CAP patients (2.95; range 0.02–10.57) (P = 0.04). Conclusions: The early induction of IP‐10 and IL‐2, as well as the subsequent over‐production of IL‐6 and lack of IL‐10 production, probably contribute to the main immuno‐pathological processes involved in lung injury in SARS. These changes in cytokine/chemokine profile are remarkably different from those observed in CAP patients.


title: Getting Unstuck: Challenges and opportunities in caring for patients experiencing prolonged hospitalization while medically ready for discharge

text: Many physicians care for patients who remain in the hospital for prolonged periods despite being ‘medical ready’ or stable for discharge. However, this phenomenon is not well defined and optimal strategies to address the problem are not known. A prolonged hospitalization past the point of medically necessity can harm patients, frustrate care teams, and is costly for the healthcare system. In this perspective, we describe opportunities to improve value of care for these patients through the lens of the Quadruple Aim, a common framework used to guide healthcare transformation efforts. We then offer recommendations, including some employed by our hospitals, for clinicians, researchers, and healthcare systems to improve the care for patients who are “stuck” in the hospital.


title: 2019_nCoV/SARS‐CoV‐2: rapid classification of betacoronaviruses and identification of Traditional Chinese Medicine as potential origin of zoonotic coronaviruses

text: The current outbreak of a novel severe acute respiratory syndrome‐like coronavirus, 2019_nCoV (now named SARS‐CoV‐2), illustrated difficulties in identifying a novel coronavirus and its natural host, as the coding sequences of various Betacoronavirus species can be highly diverse. By means of whole‐genome sequence comparisons, we demonstrate that the noncoding flanks of the viral genome can be used to correctly separate the recognized four betacoronavirus subspecies. The conservation would be sufficient to define target sequences that could, in theory, classify novel virus species into their subspecies. Only 253 upstream noncoding sequences of Sarbecovirus are sufficient to identify genetic similarities between species of this subgenus. Furthermore, it was investigated which bat species have commercial value in China, and would thus likely be handled for trading purposes. A number of coronavirus genomes have been published that were obtained from such bat species. These bats are used in Traditional Chinese Medicine, and their handling poses a potential risk to cause zoonotic coronavirus epidemics. SIGNIFICANCE AND IMPACT OF THE STUDY: The noncoding upstream and downstream flanks of coronavirus genomes allow for rapid classification of novel Betacoronavirus species and correct identification of genetic relationships. Although bats are the likely natural host of 2019_nCoV, the exact bat species that serves as the natural host of the virus remains as yet unknown. Chinese bat species with commercial value were identified as natural reservoirs of coronaviruses and are used in Traditional Chinese Medicine. Since their trading provides a potential risk for spreading zoonoses, a change in these practices is highly recommended.


title: Synthesis, Binding and Antiviral Properties of Potent Core-Extended Naphthalene Diimides Targeting the HIV-1 Long Terminal Repeat Promoter G-Quadruplexes

text: [Image: see text] We have previously reported that stabilization of the G-quadruplex structures in the HIV-1 long terminal repeat (LTR) promoter suppresses viral transcription. Here we sought to develop new G-quadruplex ligands to be exploited as antiviral compounds by enhancing binding toward the viral G-quadruplex structures. We synthesized naphthalene diimide derivatives with a lateral expansion of the aromatic core. The new compounds were able to bind/stabilize the G-quadruplex to a high extent, and some of them displayed clear-cut selectivity toward the viral G-quadruplexes with respect to the human telomeric G-quadruplexes. This feature translated into low nanomolar anti-HIV-1 activity toward two viral strains and encouraging selectivity indexes. The selectivity depended on specific recognition of LTR loop residues; the mechanism of action was ascribed to inhibition of LTR promoter activity in cells. This is the first example of G-quadruplex ligands that show increased selectivity toward the viral G-quadruplexes and display remarkable antiviral activity.


title: From single-molecule detection to next-generation sequencing: microfluidic droplets for high-throughput nucleic acid analysis

text: Droplet-based microfluidic technologies have proved themselves to be of significant utility in the performance of high-throughput chemical and biological experiments. By encapsulating and isolating reagents within femtoliter–nanoliter droplet, millions of (bio) chemical reactions can be processed in a parallel fashion and on ultra-short timescales. Recent applications of such technologies to genetic analysis have suggested significant utility in low-cost, efficient and rapid workflows for DNA amplification, rare mutation detection, antibody screening and next-generation sequencing. To this end, we describe and highlight some of the most interesting recent developments and applications of droplet-based microfluidics in the broad area of nucleic acid analysis. In addition, we also present a cursory description of some of the most essential functional components, which allow the creation of integrated and complex workflows based on flowing streams of droplets.


title: Guidelines of clinical practice for the management of swallowing disorders and recent dysphonia in the contexte of the COVID-19 pandemic

text: ABSTRACT Procedures putting healthcare workers in close contact with the airway are particularly at risk of contamination by the SARS-Cov-2 virus, especially when exposed to sputum, coughing, or a tracheostomy. In the current pandemic phase, all patients should be considered as potentially infected. Thus, the level of precaution recommended for the caregivers depends more on the type of procedure than on the patient's proved or suspected COVID-19 status. Procedures that are particularly at high risk of contamination are clinical and flexible endoscopic pharyngo-laryngological evaluation, and probably also video fluoroscopic swallowing exams. Voice rehabilitation should not be considered urgent at this time. Therefore, recommendations presented here mainly concern the management of swallowing disorders, which can sometimes be dangerous for the patient, and recent dysphonia. In cases where they are considered possible and useful, teleconsultations should be preferred to face-to-face assessments or rehabilitation sessions. The latter must be maintained only in few selected situations, after team discussions or in accordance with the guidelines provided by health authorities.


title: COVID-10 in South Korea: lessons for developing countries

text: South Korea's response is considered by many as one the most effective models against COVID-19. The average number of new cases per day fell to 6.4 in the first week of May and currently 90% of all confirmed cases have fully recovered after a massive outbreak in February 29. First, South Korea has flattened the curve of COVID-19 by combining testing, early isolation, and free treatment of positive cases combined with digital technologies without taking to “lockdown” measures. Second, South Korea has been holding press briefings twice a day and disclosed all information on COVID-19 to the public in an open and transparent manner. Third, South Korea has worked to secure the civic participation and voluntary engagement of citizens and businesses. The South Korean approach to COVID-19 may be difficult to emulate even for countries like the U.S. and U.K., but provides important policy implications for developing countries and needs for strengthening three core competencies against the outbreak of an epidemic: digital technology, efficient health governance, and civic partnership. It is a great challenge for developing countries to fight alone against COVID-19. As such, the international community should work to strengthen these core competencies accordingly.


title: Aspergillus meningitis: A rare clinical manifestation of central nervous system aspergillosis. Case report and review of 92 cases

text: Summary Objectives To describe the pathogenesis, clinical presentation, cerebrospinal fluid findings and outcome of Aspergillus meningitis, meningoencephalitis and arachnoiditis. Methods A case of Aspergillus meningitis is described. A comprehensive review of the English-language literature was conducted to identify all reported cases of Aspergillus meningitis described between January 1973 and December 2011. Results Ninety-three cases (including the one described herein) of Aspergillus meningitis were identified. Fifty-two (55.9%) were in individuals without any predisposing factor or known causes of immunosuppression. Acute and chronic meningitis was diagnosed in 65.6% of patients and meningoencephalitis in 24.7% of them with the remaining presenting with spinal arachnoiditis and ventriculitis. Cerebrospinal fluid cultures for Aspergillus spp. were positive in about 31% of cases and the galactomannan antigen test in 87%. Diagnosis during life was achieved in 52 patients (55.9%) with a case fatality rate of 50%. The overall case fatality rate was 72.1%. Conclusions Aspergillus meningitis may occur in both immunocompetent and immunocompromised patients and run an acute or chronic course. The findings of this systematic review extend the information on this life-threatening infection and could assist physicians in achieving an improved outcome.


title: The use of coiled-coil proteins in drug delivery systems

text: Abstract The coiled-coil motif is found in approximately 10% of all protein sequences and is responsible for the oligomerization of proteins in a highly specific manner. Coiled-coil proteins exhibit a large diversity of function (e.g. gene regulation, cell division, membrane fusion, drug extrusion) thus demonstrating the significance of oligomerization in biological systems. The classical coiled-coil domain comprises a series of consecutive heptad repeats in the protein sequence that are readily identifiable by the location of hydrophobic residues at the ‘a’ and ‘d’ positions. This gives rise to an α-helical structure in which between 2 and 7 helices are wound around each other in the form of a left-handed supercoil. More recently, structures of coiled-coil domains have been solved that have an 11 residue (undecad) or a 15 residue (pentadecad) repeat, which show the formation of a right-handed coiled-coil structure. The high stability of coiled coils, together with the presence of large internal cavities in the pentameric coiled-coil domain of cartilage oligomerization matrix protein (COMPcc) and the tetrameric right-handed coiled coil of Staphylothermus marinus (RHCC) has led us and others to look for therapeutic applications. In this review, we present evidence in support of a vitamin A and vitamin D3 binding activity for the pentameric COMPcc molecule. In addition, we will discuss exciting new developments which show that the RHCC tetramer is capable of binding the major anticancer drug cisplatin and the ability to fuse it to an antigenic epitope for the development of a new generation of vaccines.


title: Chapter 115 Hepatitis and Cholangiohepatitis

text: • Hepatitis is defined as any inflammatory cell infiltrate within the hepatic parenchyma; the term cholangiohepatitis describes the extension of that inflammation to include the intrahepatic bile ducts. • Although many causes of hepatitis and cholangiohepatitis have been described in dogs and cats, the cause in many cases remains unknown. • A suspicion of hepatitis or cholangiohepatitis may be based on supportive historical, physical examination, and clinicopathologic findings that are similar for most causes of hepatic disease. A diagnosis of hepatitis or cholangiohepatitis is made ultimately via histopathologic evaluation of hepatic tissue. • The mechanisms of hepatocellular injury in animals with hepatitis and cholangiohepatitis are poorly understood. Elucidation of these mechanisms may provide the basis for future therapeutic options. • Successful treatment of the patient with hepatitis or cholangiohepatitis involves addressing the underlying disease or inciting cause and providing aggressive symptomatic therapy and supportive care.


title: Mismatched Double-Stranded RNA: PolyI:PolyC12U

text: Ampligen® [polyI:polyC12U] is a mismatched double-stranded RNA that acts by inducing interferon production (immunomodulator) and by activating an intracellular enzyme (RNase-L) against viral RNA transcripts (antiviral). Ampligen®, currently under development by Hemispherx Biopharma in the US, acts on the immunological system through T-lymphocyte stimulation and is indicated for the treatment of chronic fatigue syndrome and acquired immunodeficiency deficiency syndrome (AIDS), as part of the combined therapy. Ampligen® is available for licensing worldwide. In February 2004, Fujisawa Deutschland GmbH, a subsidiary of Fujisawa Pharmaceutical Co., entered into an option agreement with Hemispherx Biopharma with the intent of becoming a distributor for Ampligen® for the potential treatment of chronic fatigue syndrome in Germany, Switzerland and Austria. An option fee of €400 000 was paid pursuant to the terms of the option agreement and upon execution of the Distribution Agreement, Fujisawa will pay Hemispherx fees and milestone payments with a potential worth of several millions of dollars.[1] In September 2003, Hemispherx Biopharma Inc. entered into an agreement with Guangdong Medicine Group Corporation to organise clinical trials, marketing, sales and distribution for both of its lead compounds, Ampligen® and Alferon N® in the People’s Republic of China. The agreement stipulates that the Guangdong Medicine Group Corporation (GMC) will conduct clinical trials with Ampligen® for the treatment of HIV. All costs related to the trials are to be covered by GMC. Additionally, GMC has to develop and implement marketing and promotional programmes.[2] In May 2003, Hemispherx Biopharma and the Center for Cell and Gene Therapy entered into a research project agreement that will see Ampligen® implemented in a protocol used in patients with relapsed EBV-positive Hodgkin’s Lymphoma.[3] In March 2002, Esteve and Hemispherx Biopharma entered into a collaborative agreement under which Esteve will be the sole distributor of Ampligen® in Spain, Portugal and Andorra for the treatment of chronic fatigue syndrome. Under this agreement, in addition to other terms, Esteve will also collaborate in the drug product development by conducting clinical studies in Spain in patients coinfected with HIV/HCV. In July 2001 Hemispherx Biopharma announced that it had formed a strategic alliance with Empire Health Resources for clinical trials of Ampligen® in the treatment of HIV and hepatitis C virus infections. Empire Health Resources, a healthcare management firm, will be responsible for accrual and retention of patients for HIV trials, and protocols for trials in patients with hepatitis C or both HIV and hepatitis C infections. Hemispherx has entered into a collaboration with RED Laboratories, and RED Laboratories NV expects that this will facilitate the continued development of Ampligen®. Hemispherx has also entered into an agreement with Schering Plough to use a Schering facility as its principal manufacturing platform in the US. This agreement may be expanded to include other territories. Hemispherx and AOP Orphan Pharmaceuticals have signed a marketing agreement for Ampligen® for the treatment of chronic fatigue syndrome for Austria, the Czech Republic, Poland and Hungary. In an arrangement between Hemispherx and Bioclones, Bioclones has certain marketing rights for Ampligen® in the Southern Hemisphere, UK and Ireland. In the US, Ampligen® has been granted orphan drug status for the treatment of AIDS, renal cell carcinoma (phase II, completed), chronic fatigue syndrome (phase III) and invasive/metastatic malignant melanoma (phase II). In August 2004, Hemispherx announced that it intends to use the proceeds from the private placement of company stock to complete the clinical work for its immunotherapeutics/ antivirals Ampligen® and Oragens™. Previously, Hemispherx submitted an application to the EMEA for the approval of Ampligen® for the treatment of chronic fatigue syndrome; the first stage of the regulatory review has been cleared. In 2000, Hemispherx Europe (Hemispherx) obtained orphan drug status for Ampligen® for the treatment of chronic fatigue syndrome in the EU, providing Hemispherx with 10 years of marketing exclusivity following the launch of the drug, as well as potential financial research benefits for the agent. In February 2000, Crystaal Corporation (now Biovail Pharmaceuticals Canada) acquired exclusive marketing rights to Ampligen® in Canada, where it submitted an NDA for the agent for the treatment of chronic fatigue syndrome. In the meantime, Ampligen® has been available since May 1996 under the Canadian Emergency Drug Release Programme for the treatment of chronic fatigue syndrome and immune dysfunction syndrome by Rivex Pharma (Helix BioPharma). Bioclones has initiated clinical studies with Ampligen® for the treatment of chronic fatigue syndrome in Australia. The active substance for Ampligen® is manufactured by F.H. Faulding Ltd. Clinical treatment programmes for chronic fatigue syndrome in other Pacific Rim countries are planned. Ampligen® is available for severe chronic fatigue syndrome on a named patient, cost-recovery basis in South Africa. Hemispherx has developed a ‘ready-to-use’ liquid formulation of the drug and has begun treating patients with chronic fatigue syndrome in ongoing clinical trials. Hemispherx has also developed an oral version of the drug (Oragen®), which is undergoing preclinical evaluation. In February 2001, Hemispherx Biopharma announced that it was initiating phase II/III trials of Ampligen® in the treatment of late-stage, multidrug-resistant strains of HIV in the European Union. Patients treated in these studies will have exhausted all other treatment options. In July 2001, Hemispherx stated that Ampligen® was being evaluated in a phase IIb trial in patients with HIV in the US. The trial, comprising two studies, REARMI and REARMII (Research/Evaluation of Ampligen® for Retroviral Mutations I and II), will evaluate the ability of Ampligen® to prevent the emergence of mutated, drug-resistant strains of the virus. ‘Several hundred’ patients currently on antiretroviral therapy and at risk of viral relapse will be enrolled at centres in Connecticut, New York, Florida and California. A second phase IIb study evaluating the effect of Ampligen® on structured treatment interruptions (STI) is also underway. Final results from this study were reported in December 2002. NIH sponsored studies of potential therapies for SARS have identified Ampligen® as having unusually high and consistent antiviral activity against human coronavirus, the pathogen implicated as the causative agent of the disease. Ampligen® demonstrated very high potency at very low concentrations (0.4 μg/mL) and had a favourable safety profile.[4] In October 2003, Hemispherx announced that, based on these promsing new results, the company will stockpile injectible and/or oral formats of Ampligen® and Alferon N®.[5] Independent researchers have demonstrated the antiviral activity of Ampligen® against flaviviruses (West Nile virus, Equine Encephalitis virus, Dengue fever virus and Japanese Encephalitis virus) as well as virus classes associated with bioterrorism. In an animal study, Ampligen® was shown to prevent destruction of nerve cells, reduce virus concentrations in the brain and blood stream and increase survival rates.[6] Researchers at the Rega Institute in Belgium have published results from an animal study demonstrating that Ampligen® was superior at protecting mice against coxsackie B3 virus-induced myocarditis compared with pegylated interferon.[7] In May 2004 Hemispherx announced that it had filed an expanded US patent application covering the use of Ampligen® for the potential treatment and prevention of severe acute respiratory syndrome (SARS) and dreaded emerging viruses.


title: Development and evaluation of a duplex real-time RT-PCR for detection and differentiation of virulent and variant strains of porcine epidemic diarrhea viruses from the United States

text: Abstract Porcine epidemic diarrhea virus (PEDV) has caused significant economic losses in the US swine industry since May 2013. A new variant strain of PEDV emerged in the US in the late December, 2013. This variant strain of PEDV differs from the virulent strain of PEDV currently circulating in the US in 1170nt of the 5’end of the S1 domain in the spike gene. Importantly, the variant PEDV caused significantly less mortality in piglets than the virulent PEDV, based on clinical observations. This suggests it may be a potential vaccine candidate for PED. Variant PEDV has been detected in samples from multiple states by our laboratory as well as other laboratories in the US. It is critical to detect and differentiate variant PEDV from the virulent PEDV during outbreaks to enhance control and to prevent PED associated disease. In this study, the development and validation of a duplex real-time RT-PCR assay for detection and differentiation of the variant and the virulent strains of PEDV currently circulating in the US was reported.


title: Lessons from SARS: A retrospective study of outpatient care during an infectious disease outbreak

text: BACKGROUND: During severe acute respiratory syndrome (SARS) outbreak in Toronto, outpatient clinics at SickKids Hospital were closed to prevent further disease transmission. In response, a decision was made by the neonatal neuro-developmental follow up (NNFU) clinic staff to select patients with scheduled appointments to have a mail/telephone assessment using Ages and Stages Questionnaire (ASQ) or to postpone/skip their visit. The objective of this study was to compare the developmental assessment and its outcome in two groups of NNFU clinic patients, SARS versus non-SARS, over three standard clinic appointments. METHODS: We compared the diagnostic accuracy (identification of developmental delay), and patient management (referral for therapy or communication of a new diagnosis) of the strategies used during SARS, April/May 2003, to the standard assessment methods used for patients seen in April/May 2005 (non-SARS). In all cases data were obtained for 3 patient visits: before, during and after these 2 months and were compared using descriptive statistics. RESULTS: There were 95 patients in the SARS group and 99 non-SARS patients. The gestational age, sex, entry diagnosis and age at the clinic visit was not different between the groups. The NNFU clinic staff mailed ASQ to 27 families during SARS, 17 (63%) were returned, and 8 of the 17 were then contacted by telephone. Criteria used to identify infants at risk selected for either mailed ASQ or phone interviews were not clearly defined in the patients' charts. There was a significant under identification of developmental delay during SARS (18% versus 45%). Of those who responded to the mailed questionnaire, referrals for therapy rates were similar to non-SARS group. The lost to follow up rate was 24% for the SARS group compared with 7% for non-SARS. There was no difference in the overall rate of developmental delay in the two groups as identified at the 'after' visit. CONCLUSIONS: Poor advanced planning led to a haphazard assessment of patients during this infectious disease outbreak. Future pandemic plans should consider planning for outpatient care as well as in hospital management of patients.


title: Improving older adults’ knowledge and practice of preventive measures through a telephone health education during the SARS epidemic in Hong Kong: A pilot study

text: Abstract Background The outbreak of severe acute respiratory syndrome (SARS) in Hong Kong posed many challenges for health promotion activities among a group of older adults with low socio-economic status (SES). With concerns that this vulnerable group could be at higher risk of contracting the disease or spreading it to others, the implementation of health promotion activities appropriate to this group was considered to be essential during the epidemic. Objectives To assess the effectiveness of delivering a telephone health education programme dealing with anxiety levels, and knowledge and practice of measures to prevent transmission of SARS among a group of older adults with low SES. Design Pretest/posttest design. Settings Subjects were recruited from registered members of a government subsidized social service center in Hong Kong and living in low-cost housing estates. Participants The eligibility criteria were: (1) aged 55 or above; (2) able to speak Cantonese; (3) no hearing impairment, and (4) reachable by telephone. Of the 295 eligible subjects, 122 older adults completed the whole study. Methods The interviewers approached all eligible subjects by telephone during the period of 15–25 May 2003. After obtaining the participants’ verbal consent, the interviewer collected baseline data by use of a questionnaire and implemented a health education programme. A follow-up telephone call was made a week later using the same questionnaire. Results The level of anxiety was lowered (t=3.28, p<0.001), and knowledge regarding the transmission routes of droplets (p<0.001) and urine and feaces (p<0.01) were improved after the intervention. Although statistical significant difference was found in the practice of identified preventive measures before and after intervention, influence on behavioral changes needed further exploration. Conclusion The telephone health education seemed to be effective in relieving anxiety and improving knowledge of the main transmission routes of SARS in this group, but not the practice of preventing SARS. Telephone contact appears to be a practical way of providing health education to vulnerable groups when face-to-face measure is not feasible and may be useful in raising health awareness during future outbreaks of emerging infections.


title: Acute disseminated encephalomyelitis in children: differential diagnosis from multiple sclerosis on the basis of clinical course

text: Acute disseminated encephalomyelitis (ADEM) is a demyelinating disease of the central nervous system (CNS) that typically presents as a monophasic disorder associated with multifocal neurologic symptoms and encephalopathy. ADEM is considered an autoimmune disorder that is triggered by an environmental stimulus in genetically susceptible individuals. The diagnosis of ADEM is based on clinical and radiological features. Most children with ADEM initially present with fever, meningeal signs, and acute encephalopathy. The level of consciousness ranges from lethargy to frank coma. Deep and subcortical white-matter lesions and gray-matter lesions such as thalami and basal ganglia on magnetic resonance imaging (MRI) are associated with ADEM. In a child who presents with signs of encephalitis, bacterial and viral meningitis or encephalitis must be ruled out. Sequential MRI is required to confirm the diagnosis of ADEM, as relapses with the appearance of new lesions on MRI may suggest either multiphasic ADEM or multiple sclerosis (MS). Pediatric MS, defined as onset of MS before the age of 16, is being increasingly recognized. MS is characterized by recurrent episodes of demyelination in the CNS separated in space and time. The McDonald criteria for diagnosis of MS include evidence from MRI and allow the clinician to make a diagnosis of clinically definite MS on the basis of the interval preceding the development of new white matter lesions, even in the absence of new clinical findings. The most important alternative diagnosis to MS is ADEM. At the initial presentation, the 2 disorders cannot be distinguished with certainty. Therefore, prolonged follow-up is needed to establish a diagnosis.


COVID-19 and the Renin Angiotensin System: Implications for the Older Adults


title: Automated assessment of COVID-19 pulmonary disease severity on chest radiographs using convolutional Siamese neural networks

text: Purpose To develop an automated measure of COVID-19 pulmonary disease severity on chest radiographs (CXRs), for longitudinal disease evaluation and clinical risk stratification. Materials and Methods A convolutional Siamese neural network-based algorithm was trained to output a measure of pulmonary disease severity on anterior-posterior CXRs (pulmonary x-ray severity (PXS) score), using weakly-supervised pretraining on ~160,000 images from CheXpert and transfer learning on 314 CXRs from patients with COVID-19. The algorithm was evaluated on internal and external test sets from different hospitals, containing 154 and 113 CXRs respectively. The PXS score was correlated with a radiographic severity score independently assigned by two thoracic radiologists and one in-training radiologist. For 92 internal test set patients with follow-up CXRs, the change in PXS score was compared to radiologist assessments of change. The association between PXS score and subsequent intubation or death was assessed. Results The PXS score correlated with the radiographic pulmonary disease severity score assigned to CXRs in the COVID-19 internal and external test sets ({rho}=0.84 and {rho}=0.78 respectively). The direction of change in PXS score in follow-up CXRs agreed with radiologist assessment ({rho}=0.74). In patients not intubated on the admission CXR, the PXS score predicted subsequent intubation or death within three days of hospital admission (area under the receiver operator characteristic curve=0.80 (95%CI 0.75-0.85)). Conclusion A Siamese neural network-based severity score automatically measures COVID-19 pulmonary disease severity in chest radiographs, which can be scaled and rapidly deployed for clinical triage and workflow optimization.


title: Wildlife health in a rapidly changing North: focus on avian disease

text: Climate‐related environmental changes have increasingly been linked to emerging infectious diseases in wildlife. The Arctic is facing a major ecological transition that is expected to substantially affect animal and human health. Changes in phenology or environmental conditions that result from climate warming may promote novel species assemblages as host and pathogen ranges expand to previously unoccupied areas. Recent evidence from the Arctic and subarctic suggests an increase in the spread and prevalence of some wildlife diseases, but baseline data necessary to detect and verify such changes are still lacking. Wild birds are undergoing rapid shifts in distribution and have been implicated in the spread of wildlife and zoonotic diseases. Here, we review evidence of current and projected changes in the abundance and distribution of avian diseases and outline strategies for future research. We discuss relevant climatic and environmental factors, emerging host–pathogen contact zones, the relationship between host condition and immune function, and potential wildlife and human health outcomes in northern regions.


title: Imported infections: What information should be collected by surveillance systems to inform public health policy?

text: Summary Background International travel carries the risk of imported diseases, which are an increasingly significant public health problem. There is little guidance about which variables should be collected by surveillance systems for strategy-based surveillance. Methods Surveillance forms for dengue, malaria, hepatitis A, typhoid and measles were collected from Australia and New Zealand and information on these compared with national surveillance forms from the UK and Canada by travel health experts. Variables were categorised by information relating to recent travel, demographics and disease severity. Results Travel-related information most commonly requested included country of travel, vaccination status and travel dates. In Australia, ethnicity information requested related to indigenous status, whilst in New Zealand it could be linked to census categories. Severity of disease information most frequently collected were hospitalisation and death. Conclusions Reviewing the usefulness of variables collected resulted in the recommendation that those included should be: overseas travel, reason for travel, entry and departure dates during the incubation period, vaccination details, traveller's and/or parents' country of birth, country of usual residence, time resident in current country, postcode, hospitalisation and death details. There was no agreement about whether ethnicity details should be collected. The inclusion of these variables on surveillance forms could enable imported infection-related policy to be formulated nationally and internationally.


title: The role of natural and synthetic zeolites as feed additives on the prevention and/or the treatment of certain farm animal diseases: A review

text: Abstract The present review comments on the role of the use of zeolites as feed additives on the prevention and/or the treatment of certain farm animal diseases. Both natural and synthetic zeolites have been used in animal nutrition mainly to improve performance traits and, based on their fundamental physicochemical properties, they were also tested and found to be efficacious in the prevention of ammonia and heavy metal toxicities, poisonings as well as radioactive elements uptake and metabolic skeletal defects. During the last decade, their utilization as mycotoxin-binding adsorbents has been a topic of considerable interest and many published research data indicate their potential efficacy against different types of mycotoxins either as a primary material or after specific modifications related to their surface properties. Ingested zeolites are involved in many biochemical processes through ion exchange, adsorption and catalysis. Recent findings support their role in the prevention of certain metabolic diseases in dairy cows, as well as their shifting effect on nitrogen excretion from urine to faeces in monogastric animals, which results in lower aerial ammonia concentration in the confinement facilities. Moreover, new evidence provide insights into potential mechanisms involved in zeolites supporting effect on animals suffered from gastrointestinal disturbances, including intestinal parasite infections. All the proposed mechanisms of zeolites’ effects are summarized in the present review and possible focus topics for further research in selected areas are suggested.


title: First characterization of a Middle-East GI-23 lineage (Var2-like) of infectious bronchitis virus in Europe

text: Abstract Variants assigned to GI-23 lineage of infectious bronchitis virus (IBV), formerly called Var2, have circulated for nearly 20 years only in countries of the Middle East. Strains of this lineage were first identified in Israel in 1998. More severe form of the virus appeared in 2006, when the second wave of Var2 epidemic has spread over the Middle East region. The present study describes the detection and detailed genetic characterization of the GI-23 viruses in Poland. The full-length genome of gammaCoV/Ck/Poland/G052/2016 strain consists of 27596 nucleotides and has typical organization for IBV (UTR5′-POl-S-3a-3b-E-M-4b-4c-5a-5b-N-UTR3′). The phylogenetic analysis of the complete sequence showed that it formed separate branch distinct from all of the full-length genome sequences analyzed in this study. Recombination analyses with other gammacoronaviruses revealed that Polish GI-23 strain may originate from recombination events and potential donors of build-in sequences are IBV of GI-1, GI-13 and G-19 lineages (Mass-, 793B- and QX-like strains, respectively). The 1a, 1b and N genes were involved in these recombination events. The source of virus introduction to the chicken population in Poland is unknown.


title: Elucidation of Cellular Targets and Exploitation of the Receptor Binding Domain of SARS‐CoV‐2 for vaccine and monoclonal antibody synthesis

text: The pandemic caused by novel severe acute respiratory syndrome coronavirus (SARS‐CoV‐2) has resulted in over 452,822 deaths in the first twenty days of June 2020 due to the coronavirus virus disease 2019 (COVID‐19). The SARS‐CoV‐2 uses the host angiotensin‐converting enzyme 2 (ACE2) receptor to gain entry inside the human cells where it replicates by using the cell protein synthesis mechanisms. The knowledge of the tissue distribution of ACE2 in human organs is therefore important to predict the clinical course of the COVID‐19. Also important is the understanding of the viral receptor‐binding domain (RBD) a region within the spike (S) proteins that enable the entry of the virus into the host cells to synthesize vaccine and monoclonal antibodies (mAbs). We performed an exhaustive search of human protein databases to establish the tissues that express ACE2 and performed an in‐depth analysis like sequence alignments and homology modeling of the spike protein (S) of the SARS‐CoV‐2 to identify antigenic regions in the RBD that can be exploited to synthesize vaccine and mAbs. Our results show that ACE2 is widely expressed in human organs that may explain the pulmonary, systemic, and neurological deficits seen in COVID‐19 patients. We show that though the S protein of the SARS‐CoV‐2 is a homolog of S protein of SARS‐CoV‐1, it has regions of dissimilarities in the RBD and transmembrane segments. We show peptide sequences in the RBD of SARS‐CoV‐2 that can bind to the MHC alleles and serve as effective epitopes for vaccine and mAbs synthesis. This article is protected by copyright. All rights reserved.


Pandemicity, COVID-19 and the limits of public health ‘science’


title: Cumulative incidence and diagnosis of SARS-CoV-2 infection in New York

text: PURPOSE: New York State (NYS) is an epicenter of the SARS-CoV-2 pandemic in the United States. Reliable estimates of cumulative incidence in the population are critical to tracking the extent of transmission and informing policies. METHODS: We conducted a statewide seroprevalence study among a 15,101 patron convenience sample at 99 grocery stores in 26 counties throughout NYS. SARS-CoV-2 cumulative incidence was estimated from antibody reactivity by first post-stratification weighting then adjusting by antibody test characteristics. The percent diagnosed was estimated by dividing diagnoses by estimated infection-experienced adults. RESULTS: Based on 1,887 of 15,101 reactive results (12.5%), estimated cumulative incidence through March 29 was 14.0% (95% CI: 13.3-14.7%), corresponding to 2,139,300 (95% CI: 2,035,800-2,242,800) infection-experienced adults. Cumulative incidence was highest in New York City (NYC) 22.7% (95% CI: 21.5-24.0%) and higher among Hispanic/Latino (29.2%), non-Hispanic black/African American (20.2%), and non-Hispanic Asian (12.4%) than non-Hispanic white adults (8.1%, p<.0001). An estimated 8.9% (95% CI: 8.4-9.3%) of infections in NYS were diagnosed, with diagnosis highest among adults ≥55 years (11.3%, 95% CI: 10.4-12.2%). CONCLUSIONS: From the largest US serosurvey to date, we estimated > 2 million adult New York residents were infected through late March, with substantial disparities, although cumulative incidence remained below herd immunity thresholds. Monitoring, testing, and contact tracing remain essential public health strategies.


title: The role of afferent pulmonary innervation in poor prognosis of acute respiratory distress syndrome in COVID-19 patients and proposed use of resiniferatoxin (RTX) to improve patient outcomes in advanced disease state: A review

text: Abstract Acute respiratory distress is one of the major causes of mortality associated with COVID-19 disease. Many patients will require intensive care with ventilatory support. Despite progress and best efforts, the mortality rates projected remain high. Historical data outlook points towards 80% expected fatality for patients progressing to advanced pulmonary disease, even when hospitalized in the intensive care unit. This is particularly true among the patient population over 65. Novel life-saving strategies are desperately needed to mitigate the high mortality that will be associated with the late stage SARS-CoV-2 viral infection associated with the fatal respiratory distress. We hypothesize that the morbidity, severity of the disease, and underlying physiological events leading to mortality are closely linked to the TRPV1 expressing neuronal system (afferent/efferent neurons) in the lungs. TRPV1 expressing cells are responsible for pain transmission, inflammation and immunomodulation throughout the entire pulmonary system and are modulating the processes associated with localized cytokine release (storm) and overall rapid disease progression. We suggest that therapeutic approaches targeting TRPV1 containing nerve fibers in the lungs will modulate the inflammatory and immune signal activity, leading to reduced mortality and better overall outcomes. We also propose to further explore the use of resiniferatoxin (RTX), an ultra-potent TRPV1 agonist currently in clinical trials for cancer and osteoarthritis pain, as a possible ablating agent of TRPV1 positive pulmonary pathways in patients with advanced COVID-19 disease.


title: An Economical Tandem Multiplex Real-Time PCR Technique for the Detection of a Comprehensive Range of Respiratory Pathogens

text: This study used real-time PCR assays to screen small sample volumes for a comprehensive range of 35 respiratory pathogens. Initial thermocycling was limited to 20 cycles to avoid competition for reagents, followed by a secondary real-time multiplex PCR. Supplementary semi-nested human metapneumovirus and picornavirus PCR assays were required to complete the acute respiratory pathogen profile. Potential pathogens were detected in 85 (70%) of pernasal aspirates collected from 121 children with acute respiratory symptoms. Multiple pathogens were detected in 29 (24%) of those samples. The tandem multiplex real-time PCR was an efficient method for the rapid detection of multiple pathogens.


title: Overview of Protein Microarrays

text: Protein microarray is an emerging technology that provides a versatile platform for characterization of hundreds of thousands of proteins in a highly parallel and high-throughput way. Two major classes of protein microarrays are defined to describe their applications: analytical and functional protein microarrays. In addition, tissue or cell lysates can also be fractionated and spotted on a slide to form a reverse-phase protein microarray. While the fabrication technology is maturing, applications of protein microarrays, especially functional protein microarrays, have flourished during the past decade. Here, we will first review recent advances in the protein microarray technologies, and then present a series of examples to illustrate the applications of analytical and functional protein microarrays in both basic and clinical research. The research areas will include detection of various binding properties of proteins, study of protein posttranslational modifications, analysis of host-microbe interactions, profiling antibody specificity, and identification of biomarkers in autoimmune diseases. As a powerful technology platform, it would not be surprising if protein microarrays will become one of the leading technologies in proteomic and diagnostic fields in the next decade.


title: Non-degradative Role of Atg5-Atg12/Atg16L1 Autophagy Protein Complex in Antiviral Activity of Interferon gamma

text: Host resistance to viral infection requires Type-I (α/β) and -II (γ) interferon (IFN) production. Another important defense mechanism is the degradative activity of macroautophagy (herein autophagy), mediated by the coordinated action of evolutionarily conserved autophagy proteins (Atg). We show that the Atg5-Atg12/Atg16L1 protein complex, whose prior known function is in autophagosome formation, is required for IFNγ-mediated host defense against murine norovirus (MNV) infection. Importantly, the direct antiviral activity of IFNγ against MNV in macrophages required Atg5-Atg12, Atg7, and Atg16L1, but not induction of autophagy, the degradative activity of lysosomal proteases, fusion of autophagosomes and lysosomes, or the Atg8 processing protein Atg4B. IFNγ, via Atg5-Atg12/Atg16L1, inhibited formation of the membranous cytoplasmic MNV replication complex, where Atg16L1 localized. Thus, the Atg5-Atg12/Atg16L1 complex performs a pivotal, nondegradative role in IFNγ-mediated antiviral defense, establishing that multicellular organisms have evolved to use portions of the autophagy pathway machinery in a cassette-like fashion for host defense.


title: Herd-level risk factors for Cryptosporidium infection in dairy-goat kids in western France

text: Abstract We conducted a cross-sectional study of risk factors for herd-level kid positivity for Cryptosporidium parvum oocysts in dairy-goat farms (Deux-Sèvres, western France). From January to March 2003, faeces from a convenient sample of 879 5- to 30-day-old goat kids from 60 herds were examined microscopically after staining with carbol fuschin. Oocyst shedding was scored semi-quantitatively (0 to 4+) allowing us to obtain a cumulative score per herd. Standardized questionnaires with information about management practices were collected in each farm. We found positive kids in 32 of 60 herds (53.3%) and in 142 animals out of 879 (16.2%). We used logistic regression for two risk-factor model: (1) simple positive (case: herd score ≥1+, at least one positive kid in the herd, versus control: herd score=0), (2) strongly positive (case: overall herd score ≥3+ versus control: herd score <3+). Risk factors associated with simple positive herds were period of sampling compared to the peak of births (After versus Before, OR=4.2, 95% CI 1.2, 15.3) and practice of kid grouping by age or weight (Yes versus No, OR=4.4, 95% CI 1.0, 19.1). Risk factors associated with strongly positive herds were period of investigation (February/March versus January, OR=12.7, 95% CI 2.1, 76.6), exposure to graminaceous plants in forage (OR=11.6, 95% CI 1.7, 81.0) and type of ventilation in the goat premises (Vertical versus Wind effect, OR=14.7, 95% CI 2.1, 106.1). No important association was found between kid-management practices and herd positivity. These results suggest a major role of the environment of kids during their first hours of life in the adult-goat premises regarding the transmission of C. parvum infection.


Retour des peurs ancestrales avec le coronavirus, Covid-19


title: Intermediate Compartment: A Sorting Station between the Endoplasmic Reticulum and the Golgi Apparatus

text: Abstract The intermediate compartment (IC) is a pleiomorphic membrane system that mediates two-way trafficing in the early biosynthetic-secretory pathway of mammalian cells. The IC associates with the cytoskeleton, binds COPI (coat protein I) coats and generates vesicular, tubular and saccular transport carriers. It recieves newly made proteins and lipids from the endoplasmic reticulum (ER) and sorts them for transport to the Golgi apparatus or recycling back to the ER. Although the IC appears to be functionally complex and resides at the crossroads of multiple transport routes, it is still disputed whether it represents a transient or stable structure.


title: Évolution de l’activité d’électro-convulsivo-thérapie en France depuis le début de la pandémie COVID-19

text: Résumé La pandémie récente de COVID-19 a entraîné des changements organisationnels majeurs dans les lieux de soins et notamment en hospitalisation en psychiatrie. Pour évaluer l’évolution de l’activité des différents centres pratiquant l’ECT, une enquête nationale en ligne a été réalisée. 65 réponses de toute la France ont été analysées. Plus de 90 % des centres pratiquant l’ECT ont connu une diminution de leur activité. Plus inquiétant encore, la moitié des centres ont subi un arrêt total de leur activité et un quart des centres accusent une diminution de plus de la moitié de leur activité habituelle. Les soins psychiatriques post-pandémie COVID-19 s’annoncent difficiles. Il est essentiel de ne pas ajouter à cette difficulté les complications, souvent graves, qui seront liées au retard ou à l’arrêt de la pratique de l’ECT. Il conviendra aussi de rester vigilant quant aux conséquences spécifiques neuropsychiatriques qui feront suite à la pandémie. Abstract The recent COVID-19 pandemic has led to major organisational changes in health care settings, especially in psychiatric hospitals. We conducted a national online survey to assess the evolution of electroconvulsive therapy (ECT) in the different centres practicing this treatment. 65 responses from all over France were analysed. More than 90 % of the centres practising ECT experienced a decrease in their activity. Half of the centres experienced a total cessation of activity and 25 % of the centres experienced a decrease of more than half of their usual activity. Post-pandemic COVID-19 psychiatric care is expected to be difficult. It is essential not to add to this difficulty the complications, often serious, that will be associated with delaying or stopping the practice of ECT. It will also be necessary to remain vigilant with regard to the specific neuropsychiatric consequences that will follow the pandemic.


title: Wie erleben Pflegefachpersonen die Corona-Pandemie?

text: Ein pflegewissenschaftliches Projekt zu Beginn der Corona-Pandemie rückt das Erleben von Pflegefachpersonen in ihrer aktuellen beruflichen Situation in den Fokus: "Warten auf die Corona-Pandemie: Wie wird die aktuelle berufliche Situation von Pflegefachpersonen in Deutschland erlebt?", lautete die Fragestellung. Über 2.000 Pflegefachpersonen aus den unterschiedlichsten Fachbereichen und Settings nahmen an der Online-Befragung teil. Die Auswertung erfolgte sowohl qualitativ wie quantitativ. Zielparameter war insbesondere das aktuelle persönliche Erleben der Teilnehmenden. Dabei gaben 95,5% der Befragten an, bisher keine vergleichbare Krisensituation erlebt zu haben. Zusatzmaterial online: Zu diesem Beitrag sind unter 10.1007/s41906-020-0761-4 für autorisierte Leser zusätzliche Dateien abrufbar.


title: ACE2, a Promising Therapeutic Target for Pulmonary Hypertension

text: Pulmonary arterial hypertension (PAH) is a chronic lung disease with poor diagnosis and limited therapeutic options. The current available therapies are ineffective in improving the quality of life and reducing mortality rates. There exists a clear unmet medical need to treat this disease, which necessitates the discovery of novel therapeutic targets/agents for safe and successful therapy. An altered renin-angiotensin system (RAS) has been implicated as a causative factor in the pathogenesis of PAH. Angiotensin II, a key effector peptide of the RAS, can exert deleterious effects on the pulmonary vasculature resulting in vasoconstriction, proliferation and inflammation, all of which contribute to PAH development. Recently, a new member of the RAS, Angiotensin converting enzyme 2 (ACE2), was discovered. This enzyme functions as a negative regulator of the angiotensin system by metabolizing Angiotensin II to a putative protective peptide, Angiotensin-(1-7). ACE2 is abundantly expressed in the lung tissue and emerging evidence suggests a beneficial role for this enzyme against lung diseases. In this review, we focus on ACE2 in relation to pulmonary hypertension and provide proof of principle for its therapeutic role in PAH.


title: Evaluation and Management of Pediatric Neck Masses An Otolaryngology Perspective

text: Most pediatric neck masses encountered in primary care are benign, reactive lymph nodes that originate from common pediatric viral processes. In a pediatric otolaryngology practice, more unusual pathologies are encountered, such as embryologic anomalies, vascular lesions, or neoplasms. Lesions that are larger or that have concerning features will ultimately need imaging and excisional biopsy for histopathologic confirmation of the diagnosis. A sound clinician understanding of anatomic neck spaces and common etiologies of pediatric neck masses can greatly reduce nonessential testing, cost, delay in treatment, and parental angst.


title: Comparison of the S genes and the biological properties of respiratory and enteropathogenic bovine coronaviruses

text: The nucleotide sequence of the S gene of the bovine respiratory coronavirus (BRCV) strain G95, which was isolated from nasal swabs of a calf suffering from respiratory disorders, was determined and compared with the S gene of the enteropathogenic bovine coronavirus (BECV) strain LY138. Sequence analysis revealed 98.7% nucleotide and 98.3% deduced amino acid identities between the S genes of BRCV-G95 and BECV-LY138 without any deletions or insertions. Nucleotide substitutions were distributed randomly throughout the gene. Five monoclonal antibodies specific for the S protein distinguished BRCV-G95 from BECV-L9, but failed to differentiate it from BECV-LY138 in Western blots under denatured and native conditions. BRCV-G95 induced cytopathic changes in cell cultures that were similar to BECV-LY138 but different from BECV-L9. These results suggest that strain BRCV-G95 is more closely related to the virulent strain BECV-LY138 than to the avirulent, cell culture-adapted strain BECV-L9.


title: Respiratory viral infections in the elderly

text: Viral respiratory infections represent a significant challenge for those interested in improving the health of the elderly. Influenza continues to result in a large burden of excess morbidity and mortality. Two effective measures, inactivated influenza vaccine, and the antiviral drugs rimantadine and amantadine, are currently available for control of this disease. Inactivated vaccine should be given yearly to all of those over the age of 65, as well as younger individuals with high-risk medical conditions and individuals delivering care to such persons. Live, intranasally administered attenuated influenza vaccines are also in development, and may be useful in combination with inactivated vaccine in the elderly. The antiviral drugs amantadine and rimantadine are effective in the treatment and prevention of influenza A, although rimantadine is associated with fewer side-effects. Recently, the inhaled neuraminidase inhibitor zanamivir, which is active against both influenza A and B viruses, was licensed for use in uncomplicated influenza. The role of this drug in treatment and prevention of influenza in the elderly remains to be determined. Additional neuraminidase inhibitors are also being developed. In addition, to influenza, respiratory infections with respiratory syncytial virus, parainfluenza virus, rhinovirus, and coronavirus have been identified as potential problems in the elderly. With increasing attention, it is probable that the impact of these infections in this age group will be more extensively documented. Understanding of the immunology and pathogenesis of these infections in elderly adults is in its infancy, and considerable additional work will need to be performed towards development of effective control measures.


Benefits of Bovine Colostrum on Fecal Quality in Recently Weaned Puppies


title: Inhibitory effects of recombinant feline interferon on the replication of feline enteropathogenic viruses in vitro

text: Abstract Antiviral activities of a recombinant feline interferon (rFeIFN) KT-80 were evaluated against feline enteropathogenic viruses in feline and canine cell lines. Sensitivity to antiviral activities of the rFeIFN varied with cell types; Felis catus whole fetus (fcwf-4) cells were more sensitive than Crandell feline kidney cells, but no sensitivity was found for Madin-Darby canine kidney cells when vesicular stomatitis virus was used as a challenge virus. Reductions were generally IFN dose-dependent and were more consistent when the cells were continuously treated with the rFeIFN than when they were pretreated only before viral challenge. Compared with each virus control culture of fcwf-4 cells, yields of rotavirus, feline panleukopenia virus (FPLV), feline calicivirus and feline infectious peritonitis coronavirus were reduced by ranges of 1.3 to ⩽3.1 log10, 0.6 log2, 0.8 to 3.7 log10 and 0.5 to 0.6 log10, respectively, in the cultures continuously treated with 10 to 10000 U of the rFeIFN. The yield reduction of FPLV was considered to be in part attributable to inhibition of cell growth by the rFeIFN supplemented in the medium.


title: Presence of Infectious Bronchitis Virus Strain CK/CH/LDL/97I in the Middle East

text: Infectious bronchitis virus (IBV) is a very dynamic and evolving virus, causing major economic losses to the global poultry industry. In early 2011, respiratory disease outbreaks were investigated in Iraq, Jordan, and Saudi Arabia. Five IBV isolates (JOA2, JOA4, Saudi-1, Saudi-2, and Iraqi IBV) were detected by diagnostic-nested nucleocapsid RT-PCR. Strain identification was characterised by sequencing and phylogenetic analysis of the amplified hypervariable region of the spike 1 (S1) gene. These five IBV isolates were found to be of the IBV strain CK/CH/LDL/97I. Nucleotide identity between these five IBV isolates ranged from 96.9% to 99.7%, and between these isolates and the CK/CH/LDL/97I strain in the range of 96.6–99.1%. The sequenced fragment of the S1 gene of the CK/CH/LDL/97I strain had less than 80% nucleotide identity to the IBV vaccine strains commonly used in the Middle East (M41 and H120). The presence of these CK/CH/LDL/97I-like strains may account for vaccination failure against IBV, since all IBV isolates were from vaccinated chickens. In this paper, we documented for the first time the presence of IBV strain CK/CH/LDL/97I in the Middle East. This strain is known to have originated in China and Taiwan.


title: Superforecasting reality check: Evidence from a small pool of experts and expedited identification

text: Superforecasting has drawn the attention of academics - despite earlier contradictory findings in the literature, arguing that humans can consistently and successfully forecast over long periods. It has also enthused practitioners, due to the major implications for improving forecast-driven decision-making. The evidence in support of the superforecasting hypothesis was provided via a 4-year project led by Tetlock and Mellers, which was based on an exhaustive experiment with more than 5000 experts across the globe, resulting in identifying 260 superforecasters. The result, however, jeopardizes the applicability of the proposition, as exciting as it may be for the academic world; if every company in the world needs to rely on the aforementioned 260 experts, then this will end up an impractical and expensive endeavor. Thus, it would make sense to test the superforecasting hypothesis in real-life conditions: when only a small pool of experts is available, and there is limited time to identify the superforecasters. If under these constrained conditions the hypothesis still holds, then many small and medium-sized organizations could identify fast and consequently utilize their own superforecasters. In this study, we provide supportive empirical evidence from an experiment with an initial (small) pool of 314 experts and an identification phase of (just) 9 months. Furthermore - and corroborating to the superforecasting literature, we also find preliminary evidence that even an additional training of just 20 minutes, can influence positively the number of superforecasters identified.


ACUTE CORONARY SYNDROME MANAGEMENT IN THE COVID-19 ERA: Voices from the Global Cardiology Community


title: Differential modulation of MHC class I antigen expression on mouse brain endothelial cells by MHV-4 infection

text: Abstract Virus-induced modulation of mouse cerebral endothelial cell class I and class II antigens by the neurotropic coronavirus, MHV-4 (JHM), was examined by flow cytometry. In susceptible BALB/c, H-2Kd was downregulated, while H-2Dd was upregulated following infection by MHV-4. In contrast, H-2K and H-2D antigens were both upregulated in either MHV-4-susceptible B10.S and (BALB/c × SJL) F1, or MHV-4-resistant SJL-derived cerebral endothelial cells following infection with this virus. Class II antigen expression was unchanged following MHV-4 infection. Virus-induced MHC class I modulation is genetically regulated, and may influence virus clearance by class I-dependent CTL.


title: US vs. Euro Area: Who drives cross-border bank lending to EMs?

text: This paper analyzes the drivers of cross-border bank lending to 49 Emerging Markets (EMs) during the period 1990Q1-2014Q4, by assessing the impact of monetary, financial and real sector shocks in both the US and the euro area. The literature has traditionally highlighted the influence of US monetary policy on driving cross-border bank flows, and more recently the importance of both US and Euro Area (EA) financial/banking sectors’ related variables. Our contribution is the simultaneous analysis of the role of these US and EA drivers, as well as their interactions with real sector shocks. We corroborate the negative impact of US monetary policy tightening on cross-border lending to EMs, but we find that EA monetary policy seems to have an impact mostly on Emerging Europe, reflecting the fact that cross-border lending to most other EM regions is dollar denominated. We also find that real sector shocks in both the US and EA trigger an increase in cross-border lending, but less in EA when modeling the financial sector. Finally, for financial sector shocks, such as those associated with a decrease in bank leverage, our results indicate a broad-based overall contraction of cross-border lending if the shock originates in the US, and heterogenous effects across borrowing regions if the shock originates in the EA.


title: Liquidity Backstops and Dynamic Debt Runs

text: Abstract Liquidity backstops can mitigate runs. In this paper we develop a dynamic model of debt runs based on He and Xiong (2012) to identify, both conceptually and quantitatively, the value of a liquidity backstop for its run-mitigating role. For the purpose of identification, we focus on the municipal bond markets for variable rate demand obligations and auction rate securities. Based on the run episodes in these markets during the financial crisis of 2007-09 and the calibrated model, we find that the value of a liquidity backstop is about 14.5 basis points per annum. Our findings have important policy implications regarding the effectiveness of liquidity backstops in ameliorating problems of financial instability.


title: Cloning and Sequence Analysis of the M gene of Porcine Epidemic Diarrhea Virus LJB/03

text: Porcine epidemic diarrhea virus (PEDV) LJB/03 was isolated from the fece of piglets infected with PEDV on a pig farm, Heilongjiang province, China. The M gene of LJB/03 was amplified from the RNA extracted directly from the fece samples by RT-PCR and cloned into pMD18-T vector. The M gene cDNA was sequenced and encompasses an open reading frame of 681 nucleotides, encoding a 226-amino acid protein. The LJB/03 M gene has a base composition of 152 adenines (22%), 153 cytosines (23%), 161 guanines (24%), and 214 thymines (31%). Sequence comparison with other PEDV strains selected from GenBank revealed that the LJB/03 M gene has a high sequence homology to those of other PEDV isolates, 97.80% with JMe2, 96.92% with KPEDV-9 (Korean field isolate), 97.36% with KPEDV-9 (Korean), 97.80% with Br1/87, and 97.94% with CV777. The encoded protein shared 97.79% amino acid identities compared with CV777, 97.35% with Br1/87, 97.79% with JMe2, 96.90% with KPEDV-9 (Korean field isolate), 96.46% with KPEDV-9 (Korean). Sequence analysis of the M gene, including genetic distance measurement, phylogenetic tree analysis, and residue substitution analysis, showed that all other PED viruses analyzed fell into three groups, and the LJB/03 itself branched in an independent group. These data revealed that the M gene nucleotide sequence of LJB/03 has some mutations in comparison with the other PED viruses.


title: Chapter 19 Nanomaterials for removal of waterborne pathogens opportunities and challenges

text: Abstract As the nanotechnological applications have taken over in different fields, their applications for water and wastewater treatment is also surfacing as a fast-developing and very promising area. Recent advancements in nanotechnological science and engineering advise that many of the waterborne pathogens could be culminated or debilitated using nanobiosorbents, nanocatalysts, bioactive nanoparticles, nanostructured catalytic membranes, nanobioreactors, nanoparticle-enhanced filtration among other products, and processes resulting from the development of nanotechnology. A detailed insight has been provided for advanced techniques such as photochemical (photocatalytic and advanced oxidation processes) applications of metal oxide nanoparticles, nanomembrane technology, bioinspired nanomaterials, and nanotechnological innovations (nano-Ag, fullerenes, nanotubes, and molecularly imprinted polymers, etc.), which prove to be highly potential as well as promising and cost-effective. However, there are still some shortcomings and challenges that must be overcome which will be looked upon in this chapter.


title: Simulation model of security control lane operation in the state of the COVID-19 epidemic

text: The article presents a discrete event simulation model for the various security control lanes configurations. The aim of the article was to check the impact of social distance on the performance of security control lanes. Different ways of passenger flow across the lane were compared. The model was verified on actual data. The results obtained indicate that it is better to use lanes with a dedicated service area for each passenger on Entry Area than a free flow along the lane. This knowledge is extremely important for security checkpoint designers. It will help ensure proper performance under normal conditions and limit performance losses under epidemic conditions. The results were also divided according to different structure of the passenger stream (percentage share of flags, lowcost and charter operations). The results showed that this has a significant impact on efficiency.


title: Serum soluble receptor for advanced glycation end‐products during acute bronchiolitis in infant: Prospective study in 93 cases

text: INTRODUCTION: Acute bronchiolitis is a major cause of acute respiratory distress in infants. The soluble receptor for advanced glycation end‐products (sRAGE) is a biomarker of pulmonary damage processes, with a diagnostic and a prognostic value in acute respiratory distress syndrome (ARDS). The RAGE pathway is also implicated in the pathogenesis of other respiratory diseases like asthma, but the value of sRAGE levels in acute bronchiolitis remains under‐investigated. MATERIAL AND METHODS: A prospective, observational, and analytical study was conducted at Clermont‐Ferrand University Hospital. The main objective was to evaluate the correlation between serum sRAGE and clinical severity of bronchiolitis in hospitalized infants aged <1 year. We analyzed correlations between serum sRAGE and Wainwright score, short‐term morbidity attributable to bronchiolitis, causal viruses and risk for recurrent wheezing at 1 year. RESULTS: The study included 93 infants. sRAGE levels were significantly lower in acute bronchiolitis patients (mean 1101 pg/mL) than in controls (2203 pg/mL, P < 0.001) but did not correlate with clinical severity. No correlation was found between serum sRAGE and severity score, respiratory viruses, and recurrent wheezing at 1 year. Serum sRAGE levels were negatively correlated with age (r = −0.45, P < 0.001). CONCLUSION: Serum sRAGE levels are decreased in acute bronchiolitis but not correlated with disease severity. sRAGE levels should be age‐adjusted in infants. Serum sRAGE levels measured in the setting of acute bronchiolitis were not predictive of recurrent wheezing.