title: Computational analysis and determination of a highly conserved surface exposed segment in H5N1 avian flu and H1N1 swine flu neuraminidase

text: BACKGROUND: Catalytic activity of influenza neuraminidase (NA) facilitates elution of progeny virions from infected cells and prevents their self-aggregation mediated by the catalytic site located in the body region. Research on the active site of the molecule has led to development of effective inhibitors like oseltamivir, zanamivir etc, but the high rate of mutation and interspecies reassortment in viral sequences and the recent reports of oseltamivir resistant strains underlines the importance of determining additional target sites for developing future antiviral compounds. In a recent computational study of 173 H5N1 NA gene sequences we had identified a 50-base highly conserved region in 3'-terminal end of the NA gene. RESULTS: We extend the graphical and numerical analyses to a larger number of H5N1 NA sequences (514) and H1N1 swine flu sequences (425) accessed from GenBank. We use a 2D graphical representation model for the gene sequences and a Graphical Sliding Window Method (GSWM) for protein sequences scanning the sequences as a block of 16 amino acids at a time. Using a protein sequence descriptor defined in our model, the protein sliding scan method allowed us to compare the different strains for block level variability, which showed significant statistical correlation to average solvent accessibility of the residue blocks; single amino acid position variability results in no correlation, indicating the impact of stretch variability in chemical environment. Close to the C-terminal end the GSWM showed less descriptor-variability with increased average solvent accessibility (ASA) that is also supported by conserved predicted secondary structure of 3' terminal RNA and visual evidence from 3D crystallographic structure. CONCLUSION: The identified terminal segment, strongly conserved in both RNA and protein sequences, is especially significant as it is surface exposed and structural chemistry reveals the probable role of this stretch in tetrameric stabilization. It could also participate in other biological processes associated with conserved surface residues. A RNA double hairpin secondary structure found in this segment in a majority of the H5N1 strains also supports this observation. In this paper we propose this conserved region as a probable site for designing inhibitors for broad-spectrum pandemic control of flu viruses with similar NA structure.


title: Seasonality and prevalence of respiratory pathogens detected by multiplex PCR at a tertiary care medical center

text: Respiratory tract infections (RTIs) are a leading cause of mortality and morbidity. Seasonality has been reported for many viruses, including influenza virus, respiratory syncytial virus (RSV), and the recently described human metapneumovirus (hMPV). We hypothesize that the availability of rapid, multiplex PCR diagnostics will provide better clinical care and new insights into the etiology and clinical spectrum of RTIs. We conducted a retrospective analysis of the incidence of respiratory pathogens at a 500-bed adult and 154-bed pediatric hospital tertiary care center. A total of 939 specimens from patients with an age range of 5 days to 91 years (median, 2 years) were tested by a multiplex respiratory pathogen PCR from November 14, 2011 to November 13, 2012. Sixty-five percent of specimens were positive for at least one pathogen. As the age of the patient increased, the positivity rate for the PCR decreased proportionately. Rhinoviruses/enteroviruses (Rhino/Entero) were the most prevalent (34.3 %) followed by RSV (19.2 %) and hMPV (6.2 %). Twelve percent of the positive samples were positive for multiple analytes, with Rhino/Entero and RSV being the most common combination. The peak months were September and May for Rhino/Entero infections, January for RSV and February for coronavirus. hMPV peaked 2 months after RSV, as has been observed recently in other studies. Multiplex PCR provides rapid diagnostic information that can be used to make knowledgeable clinical decisions and potentially reduce the use of antibiotics. Active respiratory PCR surveillance could also predict seasonal respiratory epidemics to allow for adequate planning of additional infection control measures.


title: The substrate specificity of SARS coronavirus 3C-like proteinase

text: Abstract The 3C-like proteinase of severe acute respiratory syndrome coronavirus (SARS) has been proposed to be a key target for structural based drug design against SARS. We have designed and synthesized 34 peptide substrates and determined their hydrolysis activities. The conserved core sequence of the native cleavage site is optimized for high hydrolysis activity. Residues at position P4, P3, and P3′ are critical for substrate recognition and binding, and increment of β-sheet conformation tendency is also helpful. A comparative molecular field analysis (CoMFA) model was constructed. Based on the mutation data and CoMFA model, a multiply mutated octapeptide S24 was designed for higher activity. The experimentally determined hydrolysis activity of S24 is the highest in all designed substrates and is close to that predicted by CoMFA. These results offer helpful information for the research on the mechanism of substrate recognition of coronavirus 3C-like proteinase.


title: Chapter 2 Ubiquitous waterborne pathogens

text: Abstract Water is one of the most important substances on earth and without it life cannot exist. However, poor water quality in many parts of the world has increased the number of water-related diseases, making it the leading cause of disease and death globally for both young and old. Waterborne pathogens cause diseases in humans through two major exposure pathways: drinking water and recreational waters. This chapter on waterborne pathogens will be starting with an introduction, followed by descriptions on classical waterborne pathogens; bacteria, viruses, protozoans, and helminths placing emphasis on the World Health Organization guidelines. Further to conventional waterborne pathogens, fresh organisms and new strains from already known pathogens are being identified and that present important additional challenges to both the water and public health sectors. Hence later part of the chapter focuses on the potential waterborne pathogens and will conclude with a summary of the content.


title: Randomized controlled clinical trial on the effect of oral immunoglobulin supplementation on neonatal dairy calves with diarrhea

text: BACKGROUND: Nonantibiotic alternatives providing local gut immunity have been recommended for managing calf diarrhea. ANIMALS: One hundred and two calves with diarrhea. HYPOTHESIS: Oral supplementation with immunoglobulins in calves with diarrhea will reduce time to resolution of diarrhea, number of treatment events, and mortality rate. METHODS: Randomized controlled trial. Calves were assigned into 1 of 3 groups. The treatment group was supplemented with 20 g of immunoglobulins in milk twice daily for 14 days. The placebo group was supplemented with 20 g of a product with similar nutritional value as the treatment group, but without immunoglobulins, in milk, twice daily for 14 days. The control group received no supplements. Medical treatments, time to resolution of diarrhea, and case fatality rates were compared. RESULTS: There was no difference in the proportion of treatment events (treatment, 79% versus placebo, 77% versus control, 71%) among groups (P = .69). The median time to resolution of diarrhea was not different between the treatment (10.5 days; 95% confidence interval [CI], 7, 13) and control (8 days; 95% CI, 5, 10) groups (P = .08) or between the placebo (6.5 days; 95% CI, 3, 9) and control groups (P = .89). Median time to resolution was shorter (P = .008) in the placebo compared to the treatment group (6.5 versus 10.5 days). Case fatality rates among groups (treatment, 12% versus placebo, 3% versus control, 3%) were not different (P = .36). CONCLUSIONS AND CLINICAL IMPORTANCE: Expected benefits of conferring local gut immunity by immunoglobulin supplementation in calves with diarrhea were not evident.


Protection protocol in intubation of suspected SARS patients


title: Mycoplasma pneumoniae-Associated Bronchiolitis Causing Severe Restrictive Lung Disease in Adults Report of Three Cases and Literature Review

text: Study objectives To characterize adult Mycoplasma pneumoniae-induced bronchiolitis requiring hospitalization. Design We encountered an adult patient with severe bronchiolitis in the absence of pneumonia due to M pneumoniae. To determine the relative frequency of such a condition, we retrospectively reviewed the medical records of adults over a 4-year period with a hospital discharge diagnosis of “bronchiolitis” from a university hospital. Setting University Hospital of the University of Colorado Health Sciences Center, Denver, CO. Study subjects From 1994 to 1998, 10 adult inpatients were identified with a diagnosis of bronchiolitis. There were two with respiratory bronchiolitis, one with panbronchiolitis, one patient with bronchiolitis obliterans organizing pneumonia (BOOP), and six with acute inflammatory bronchiolitis. Including the initial patient, three had a definitive clinical diagnosis of Mycoplasma-associated bronchiolitis. Results The three adult patients with bronchiolitis due to M pneumoniae are unusual because they occurred in the absence of radiographic features of a lobar or patchy alveolar pneumonia. Hospital admission was occasioned by the severity of symptoms and gas exchange abnormalities. One patient had bronchiolitis as well as organizing pneumonia (BOOP) that responded favorably to corticosteroid treatment. The other two had high-resolution CT findings diagnostic of an acute inflammatory bronchiolitis. One of the patients with inflammatory bronchiolitis had an unusual pattern of marked ventilation and perfusion defects localized predominantly to the left lung. All three had restrictive ventilatory impairment on physiologic testing. Conclusions In adults, Mycoplasma-associated bronchiolitis without pneumonia is rarely reported, but in hospitalized patients, it may be more common than expected and may be associated with severe physiologic disturbances.


title: Co-circulating genetically divergent A2 human metapneumovirus strains among children in southern Taiwan

text: An outbreak of human metapneumovirus (hMPV) among children in southern Taiwan in 2004 prompted the investigation of the molecular epidemiology of hMPV from September 2003 to August 2005. Respiratory specimens that were culture negative for a panel of respiratory viruses were examined for the presence of hMPV by RT-PCR. The results indicated that 59 out of 546 (10.8%) children were hMPV-positive. The majority of these hMPV-positive children were less than 2 years old (59.4%), females (61%), and inpatients (67.8%). Infections occurred throughout the year, but peaked during the spring and/or summer months. Sequence analysis of the fusion gene from the isolates revealed two phylogenetic groups with five possible lineages (A1, A2a/A2b, B1, and B2). Among these co-circulating strains, A2 strains were most frequently observed and demonstrated the greatest divergence. Deduced amino acid sequence analysis identified several variant amino acids specific to the A2 lineage. Lineage-specific amino acid substitutions were noted at aa233, aa286, aa312, aa348, and aa296. This study indicated that genetically divergent strains of hMPV which caused respiratory disease and hospitalization were circulating among children in Taiwan.


title: Identification of an endocytic signal essential for the antiviral action of IFITM3

text: Members of the interferon-induced transmembrane (IFITM) protein family inhibit the entry of a wide range of viruses. Viruses often exploit the endocytosis pathways to invade host cells and escape from the endocytic vesicles often in response to low pH. Localization to these endocytic vesicles is essential for IFITM3 to interfere with the cytosolic entry of pH-dependent viruses. However, the nature of the sorting signal that targets IFITM3 to these vesicles is poorly defined. In this study, we report that IFITM3 possesses a YxxΦ sorting motif, i.e., 20-YEML-23, that enables IFITM3 to undergo endocytosis through binding to the μ2 subunit of the AP-2 complex. IFITM3 accumulates at the plasma membrane as a result of either mutating 20-YEML-23, depleting the μ2 subunit, or overexpressing μ2 mutants. Importantly, blocking endocytosis of IFITM3 abrogates its ability to inhibit pH-dependent viruses. We have therefore identified a critical sorting signal, namely 20-YEML-23, that controls both the endocytic trafficking and the antiviral action of IFITM3. This finding also reveals that as an endocytic protein, IFITM3 first arrives at the plasma membrane before it is endocytosed and further traffics to the late endosomes where it acts to impede virus entry.


title: Evolution of CT findings in patients with mild COVID-19 pneumonia

text: OBJECTIVES: To delineate the evolution of CT findings in patients with mild COVID-19 pneumonia. METHODS: CT images and medical records of 88 patients with confirmed mild COVID-19 pneumonia, a baseline CT, and at least one follow-up CT were retrospectively reviewed. CT features including lobar distribution and presence of ground glass opacities (GGO), consolidation, and linear opacities were analyzed on per-patient basis during each of five time intervals spanning the 3 weeks after disease onset. Total severity scores were calculated. RESULTS: Of patients, 85.2% had travel history to Wuhan or known contact with infected individuals. The most common symptoms were fever (84.1%) and cough (56.8%). The baseline CT was obtained on average 5 days from symptom onset. Four patients (4.5%) had negative initial CT. Significant differences were found among the time intervals in the proportion of pulmonary lesions that are (1) pure GGO, (2) mixed attenuation, (3) mixed attenuation with linear opacities, (4) consolidation with linear opacities, and (5) pure consolidation. The majority of patients had involvement of ≥ 3 lobes. Bilateral involvement was more prevalent than unilateral involvement. The proportions of patients observed to have pure GGO or GGO and consolidation decreased over time while the proportion of patients with GGO and linear opacities increased. Total severity score showed an increasing trend in the first 2 weeks. CONCLUSIONS: While bilateral GGO are predominant features, CT findings changed during different time intervals in the 3 weeks after symptom onset in patients with COVID-19. KEY POINTS: • Four of 88 (4.5%) patients with COVID-19 had negative initial CT. • Majority of COVID-19 patients had abnormal CT findings in ≥ 3 lobes. • A proportion of patients with pure ground glass opacities decreased over the 3 weeks after symptom onset.


title: Detection of SARS-COV-2 in Placental and Fetal Membrane Samples

text: Introduction Since the first reports of the emergence of the novel coronavirus SARS-CoV-2 and its associated disease (COVID-19), concerns remain about whether the virus is transmissible in pregnant women from the mother to the fetus during either the antepartum period or the process of labor and delivery. One recent review reported that in a small number of cases, two PCR swabs of the placenta were sent in additional to neonatal and cord blood testing, and both placental PCR swabs were negative.1 Other studies have demonstrated the finding of SARS-CoV2 IgM in neonates born to mothers diagnosed with COVID-19 during pregnancy,2,3 findings that may indicate vertical transmission of the virus in utero. We report our experience with placental/membrane SARS-CoV2 RNA PCR swab results after delivery to a series of symptomatic mothers with confirmed COVID-19 infection in pregnancy. Methods IRB approval was obtained. All pregnant patients diagnosed with COVID-19 who gave birth between March 1, 2020 and April 20, 2020 at NYU Langone Health were identified by a search of the electronic medical record. Charts were reviewed for documentation of SARS-CoV-2 RNA RT-PCR testing sent from either the placenta or membranes within 30 minutes following delivery. PCR testing for SARS-COV-2 was performed using the cobas SARS-CoV-2 assay (Roche) or the Cepheid Xpert Xpress assay. Placental swabs were obtained from the amniotic surface after clearing the surface of maternal blood (placental PCR). Membrane swabs were obtained from between the amnion and chorion after manual separation of the membranes (membrane PCR). Maternal COVID-19 illness was categorized as mild, severe, or critical.4 The time interval from maternal diagnosis of COVID-19 to delivery was calculated in days. Infants were tested with nasopharyngeal swabs for SARS-CoV-2 PCR between days of life 1 and 5 while hospitalized. Hospitalized infants were also assessed for clinical signs and symptoms, including fever, cough, and nasal congestion. Results Of 32 COVID-19 positive pregnant patients who gave birth in this timeframe, placental or membrane swabs were sent from 11 patients (Table). Three of 11 swabs were positive. None of the infants tested positive for SARS-CoV2 on days of life 1 through 5, and none demonstrated symptoms of COVID-19 infection. Discussion Of 11 placental or membrane swabs sent following delivery, 3 swabs were positive for SARS-CoV-2, all in women with moderate to severe COVID-19 illness at time of delivery. This is the first study to demonstrate the presence of SARS-CoV-2 RNA in placental or membrane samples. While there were no clinical signs of vertical transmission, our findings raise the possibility of intrapartum viral exposure. Given the mixing of maternal and fetal fluid and tissue at time of delivery, the origin of the detected SARS-CoV-2 RNA in our series is unclear. It may represent contamination from maternal blood, amniotic fluid, or COVID-19 infection of the membranes and amniotic sac. For those infants who were delivered vaginally, contamination with vaginal secretions is also a possible source, although prior studies on vaginal secretions have failed to demonstrate the presence of COVID-19.5,6 Although all of our neonates tested negative in the first 5 days of life, many were born via cesarean deliveries with decreased length of exposure to these tissues, which may be associated with a decreased likelihood of vertical transmission. Additionally, nasopharyngeal testing immediately after delivery may not be the ideal approach to evaluate vertical transmission if exposure occurs at the time of delivery, as the virus may require a longer incubation period before these swabs convert to positive. In summary, the presence of viral RNA by RT-PCR in placenta/membranes at the time of delivery suggests the need for further research into the possibility of vertical transmission.


title: Rapid point of care diagnostic tests for viral and bacterial respiratory tract infections—needs, advances, and future prospects

text: Summary Respiratory tract infections rank second as causes of adult and paediatric morbidity and mortality worldwide. Respiratory tract infections are caused by many different bacteria (including mycobacteria) and viruses, and rapid detection of pathogens in individual cases is crucial in achieving the best clinical management, public health surveillance, and control outcomes. Further challenges in improving management outcomes for respiratory tract infections exist: rapid identification of drug resistant pathogens; more widespread surveillance of infections, locally and internationally; and global responses to infections with pandemic potential. Developments in genome amplification have led to the discovery of several new respiratory pathogens, and sensitive PCR methods for the diagnostic work-up of these are available. Advances in technology have allowed for development of single and multiplexed PCR techniques that provide rapid detection of respiratory viruses in clinical specimens. Microarray-based multiplexing and nucleic-acid-based deep-sequencing methods allow simultaneous detection of pathogen nucleic acid and multiple antibiotic resistance, providing further hope in revolutionising rapid point of care respiratory tract infection diagnostics.


title: An Alert to COVID-19 Literature in Predatory Publishing Venues

text: Abstract The COVID-19 pandemic, which has led to a flood of papers and preprints, has placed multiple challenges on academic publishing, the most obvious one being sustained integrity under the pressure to publish quickly. There are risks of this high volume-to-speed ratio. Many letters, editorials, and supposedly “peer reviewed” papers in ranked and indexed journals were published in a matter of days, suggesting that peer review was either fleeting or non-existential, or that papers were rapidly approved by editors based on their perceived interest and topicality, rather than on their intrinsic academic value. In academic publishing circles, the claim of “peer review”, when in fact it has not been conducted, is a core characteristic of “predatory publishing”, and is also a “fake” element that may undermine efforts in recent years to build trust in science's budding serials crisis. While the world is still centrally focused on COVID-19, the issue of “predatory publishing” is being ignored, or not being given sufficient attention. The risks to the scholarly community, academic publishing and ultimately public health are at stake when exploitative and predatory publishing are left unchallenged.


title: Healthcare impact of COVID-19 epidemic in India: A stochastic mathematical model

text: Abstract Background In India, the SARS-CoV-2 COVID-19 epidemic has grown to 1251 cases and 32 deaths as on 30 Mar 2020. The healthcare impact of the epidemic in India was studied using a stochastic mathematical model. Methods A compartmental SEIR model was developed, in which the flow of individuals through compartments is modeled using a set of differential equations. Different scenarios were modeled with 1000 runs of Monte Carlo simulation each using MATLAB. Hospitalization, intensive care unit (ICU) requirements, and deaths were modeled on SimVoi software. The impact of nonpharmacological interventions (NPIs) including social distancing and lockdown on checking the epidemic was estimated. Results Uninterrupted epidemic in India would have resulted in more than 364 million cases and 1.56 million deaths with peak by mid-July. As per the model, at current growth rate of 1.15, India is likely to reach approximately 3 million cases by 25 May, implying 125,455 (±18,034) hospitalizations, 26,130 (±3298) ICU admissions, and 13,447 (±1819) deaths. This would overwhelm India's healthcare system. The model shows that with immediate institution of NPIs, the epidemic might still be checked by mid-April 2020. It would then result in 241,974 (±33,735) total infections, 10,214 (±1649) hospitalizations, 2121 (±334) ICU admissions, and 1081 (±169) deaths. Conclusion At the current growth rate of epidemic, India's healthcare resources will be overwhelmed by the end of May. With the immediate institution of NPIs, total cases, hospitalizations, ICU requirements, and deaths can be reduced by almost 90%.


title: A clustering of rna recombination sites adjacent to a hypervariable region of the peplomer gene of murine coronavirus

text: Abstract Coronaviruses undergo RNA recombination at a very high frequency. To understand the mechanism of recombination in murine coronavirus, we have performed RNA sequencing of viral genomic RNA to determine the precise sites of recombination in a series of recombinants which have crossovers within the gene encoding the peplomer protein. We found that all of the recombination sites are clustered within a region of 278 nucleotides in the 5′-half of the gene. This region in which all of the crossovers occurred represents a small fraction of the distance between the two selection markers used for the isolation of these recombinant viruses. This result suggests that this region may be a preferred site for RNA recombination. The crossover sites are located within and immediately adjacent to a hypervariable area of the gene. This area has undergone deletions of varying sizes in several virus strains which have been passaged either in vivo or in vitro. These results suggest that a similar RNA structure may be involved in the occurrence of both recombination and deletion events.


title: In Silico Design of Antiviral Peptides Targeting the Spike Protein of SARS-CoV-2

text: An outbreak caused by 2019 novel coronavirus (2019-nCoV) was first identified in Wuhan City, Hubei Province, China. The new virus was later named SARS-CoV-2. The virus has affected tens of thousands of patients in the world. The infection of SARS-CoV-2 causes severe pneumonia and even death. It is urgently needed to find a therapeutic method to treat patients with SARS-CoV-2 infection. Studies showed that the surface spike (S) protein is essential for the coronavirus binding and entry of host cells. The heptad repeats 1 and 2 (HR1 and HR2) in the S protein play a decisive role in the fusion of the viral membrane with the host cell membrane. We predicted the HR1 and HR2 regions in S protein by sequence alignment. We simulated a computational model of HR1/2 regions and the fusion core. The binding energy of HR1 and HR2 of the fusion core was -33.4kCal/mol. We then designed antivirus peptides by molecular dynamics simulation of the fusion core. The binding energy of HR2-based antiviral peptide to HR1 was -43.0kCal/mol, which was stronger than the natural stage of the fusion core, suggesting that the predicted antiviral peptide can competitively bind with HR1 to prevent forming of the fusion core. The antiviral peptides can prevent SARS-CoV-2 membrane fusion and can potentially be used for the prevention and treatment of infections.


title: Interleukin-1 receptor antagonist anakinra in association with remdesivir in severe Coronavirus disease 2019: A case report

text: Abstract We report the first successful treatment with the IL-1 receptor antagonist anakinra, in association with the most promising and available antiviral therapy, of a severe case of novel coronavirus disease 2019 (COVID-19). We describe the diagnosis, clinical course, and management of the case, including the respiratory failure at presentation, the progression to a scenario characterized by profound inflammatory dysregulation similar to that observed during macrophage activation syndrome, and the clinical improvement after treatment with the IL-1 receptor antagonist anakinra. This case highlights the high tolerability and the interesting immunomodulatory profile of the IL-1 receptor antagonist anakinra in the setting of severe COVID-19 associated with remdesivir therapy. Further studies are needed to confirm the safety and efficacy of this combination strategy in the treatment of this emerging infection.


title: People at Risk of Influenza Pandemics: The Evolution of Perception and Behavior

text: Influenza pandemics can severely impact human health and society. Understanding public perception and behavior toward influenza pandemics is important for minimizing the effects of such events. Public perception and behavior are expected to change over the course of an influenza pandemic, but this idea has received little attention in previous studies. Our study aimed to understand the dynamics of public perception and behavior over the course of the 2009 H1N1 influenza pandemic. Three consecutive cross-sectional surveys were administered among Beijing residents with random-digit dialing techniques in March 2008 and August and November 2009. Effective samples of 507, 508 and 1006 respondents were interviewed in each of the three surveys, respectively. The mean scores of risk perception were low to moderate across the three surveys. The perceived risk of infection of self was significantly lower than that of the community, revealing an optimistic bias. Longitudinally, the perceived risk of contracting H1N1 increased, whereas the perceived risk of being unable to obtain medicine and medical care once influenza permeated the community first increased and then decreased. Responsive actions toward influenza varied. Most respondents took actions that required little extra effort, such as ventilating rooms; these actions did not change over time. Comparatively, a smaller number of respondents took actions for coping with influenza, such as vaccination; however, these actions were taken by an increasing number of respondents over time. The association between risk perception and behavior was unstable. Positive, insignificant, and negative associations were obtained in the three surveys. In conclusion, the evolving patterns of risk perception and responsive behavior over the course of an influenza pandemic are sensitive to how risk and behavior are defined and scoped.


Biologika und Covid-19


title: Longitudinal anti-SARS-CoV-2 antibody profile and neutralization activity of a COVID-19 patient

text: We followed–up a mild COVID-19 patient for 91 days and serially monitored his serum antibodies to four SARS-CoV-2 related antigens (NP, RBD, S1 and ECD) and neutralization activities. Our data revealed a profile of serial antibody responses during the progress and a quick decline of neutralization activities after discharge.


title: GPI-anchored single chain Fv - an effective way to capture transiently-exposed neutralization epitopes on HIV-1 envelope spike

text: BACKGROUND: Identification of broad neutralization epitopes in HIV-1 envelope spikes is paramount for HIV-1 vaccine development. A few broad neutralization epitopes identified so far are present on the surface of native HIV-1 envelope spikes whose recognition by antibodies does not depend on conformational changes of the envelope spikes. However, HIV-1 envelope spikes also contain transiently-exposed neutralization epitopes, which are more difficult to identify. RESULTS: In this study, we constructed single chain Fvs (scFvs) derived from seven human monoclonal antibodies and genetically linked them with or without a glycosyl-phosphatidylinositol (GPI) attachment signal. We show that with a GPI attachment signal the scFvs are targeted to lipid rafts of plasma membranes. In addition, we demonstrate that four of the GPI-anchored scFvs, but not their secreted counterparts, neutralize HIV-1 with various degrees of breadth and potency. Among them, GPI-anchored scFv (X5) exhibits extremely potent and broad neutralization activity against multiple clades of HIV-1 strains tested. Moreover, we show that GPI-anchored scFv (4E10) also exhibited more potent neutralization activity than its secretory counterpart. Finally, we demonstrate that expression of GPI-anchored scFv (X5) in the lipid raft of plasma membrane of human CD4(+ )T cells confers long-term resistance to HIV-1 infection, HIV-1 envelope-mediated cell-cell fusion, and the infection of HIV-1 captured and transferred by human DCs. CONCLUSIONS: Thus GPI-anchored scFv could be used as a general and effective way to identify antibodies that react with transiently-exposed neutralization epitopes in envelope proteins of HIV-1 and other enveloped viruses. The GPI-anchored scFv (X5), because of its breadth and potency, should have a great potential to be developed into anti-viral agent for HIV-1 prevention and therapy.


title: Spiro‐Indole‐Coumarin Hybrids: Synthesis, ADME, DFT, NBO Studies and In Silico Screening through Molecular Docking on DNA G‐Quadruplex

text: New series of hybrids were synthesized by combination of 4‐hydroxycoumarin with spiro[indol‐indazole‐thiazolidine]‐diones and spiro[indol‐pyrazole‐thiazolidine]‐diones, via hitherto unknown Schiff bases. The effects of substituents, such as ‐F, ‐Br and ‐CH(3), on the crucial characteristics pertaining to the hybrids were investigated through computational studies. In silico or virtual screening through molecular docking studies on the library of 22 compounds, including reference compounds, precursors, non‐hybrid and hybrid derivatives, was performed on DNA G‐quadruplex of the human genome. All six freshly synthesized hybrids showed high binding energy as compared to non‐hybrids as well as reference compounds. The presence of substituents at 5‐position of indole enhanced the binding tendency of the ligand. ADME studies indicated good oral bioavailability and absorption of these compounds. Density Functional Theory (DFT) calculations of hybrids were done at B3LYP/6‐311G++(d,p) level of computation. Their HOMO and LUMO energy plots reflected the presence of high charge transfer and chemical potential. Natural bond order (NBO) calculations predicted hyperconjugative interactions. The Molecular Electrostatic Potential (MEP) surface plots showed possible electrophilic and nucleophilic attacking sites of the hybrids. Compound 10 a (5‐fluoro‐spiro[indol‐indazole‐thiazolidine]‐dione‐coumarin hybrid), on the basis of global reactivity descriptors, was filtered to be chemically most reactive with the highest binding energy of −8.23 kcal/mol with DNA G‐quadruplex. The synthesized hybrid coumarin derivatives in correlation with theoretical docking studies validate that hybrid derivatives are more reactive compared to their non‐hybrid counterparts.


title: Complete Genome Viral Phylogenies Suggests the Concerted Evolution of Regulatory Cores and Accessory Satellites

text: We consider the concerted evolution of viral genomes in four families of DNA viruses. Given the high rate of horizontal gene transfer among viruses and their hosts, it is an open question as to how representative particular genes are of the evolutionary history of the complete genome. To address the concerted evolution of viral genes, we compared genomic evolution across four distinct, extant viral families. For all four viral families we constructed DNA-dependent DNA polymerase-based (DdDp) phylogenies and in addition, whole genome sequence, as quantitative descriptions of inter-genome relationships. We found that the history of the polymerase gene was highly predictive of the history of the genome as a whole, which we explain in terms of repeated, co-divergence events of the core DdDp gene accompanied by a number of satellite, accessory genetic loci. We also found that the rate of gene gain in baculovirus and poxviruses proceeds significantly more quickly than the rate of gene loss and that there is convergent acquisition of satellite functions promoting contextual adaptation when distinct viral families infect related hosts. The congruence of the genome and polymerase trees suggests that a large set of viral genes, including polymerase, derive from a phylogenetically conserved core of genes of host origin, secondarily reinforced by gene acquisition from common hosts or co-infecting viruses within the host. A single viral genome can be thought of as a mutualistic network, with the core genes acting as an effective host and the satellite genes as effective symbionts. Larger virus genomes show a greater departure from linkage equilibrium between core and satellites functions.


title: Radiographic and ultrasonographic findings of the spleen and abdominal lymph nodes in healthy domestic ferrets

text: OBJECTIVE: To describe the radiographic and ultrasonographic characteristics of the spleen and abdominal lymph nodes in clinically healthy ferrets. MATERIALS AND METHODS: Fifty‐five clinically healthy ferrets were prospectively recruited for this cross‐sectional study. Three‐view whole body radiographs and abdominal ultrasonography were performed on awake (23 out of 55) or sedated (32 out of 55) ferrets. On radiographs splenic and abdominal lymph node visibility was assessed. Splenic thickness and echogenicity and lymph node length, thickness, echogenicity, number and presence of cyst‐like changes were recorded. RESULTS: The spleen was radiographically detectable in all ferrets. On ultrasound the spleen was hyperechoic to the liver (55 out of 55) and mildly hyperechoic (28 out of 55), isoechoic (15 out of 55) or mildly hypoechoic (12 out of 55) to the renal cortices. Mean splenic thickness was 11.80 ±0.34 mm. Lymph nodes were radiographically discernible in 28 out of 55 ferrets and included caudal mesenteric and sublumbar nodes. An average of 9 ±2 lymph nodes (mean± standard deviation; mode 10) were identified in each ferret using ultrasound. A single large jejunal lymph node was identified in all ferrets and had a mean thickness of 5.28 ± 1.66 mm. For other lymph nodes the mean thickness measurements plus one standard deviation were less than 4.4 mm (95% confidence interval: ≤ 3.72 mm). CLINICAL SIGNIFICANCE: The information provided in this study may act as a baseline for evaluation of the spleen and lymph nodes in ferrets.


title: Helix-helix interactions inside lipid bilayers

text: Abstract Far from being simple hydrophobic anchors, it is now clear that the transmembrane α-helices of integral membrane proteins can participate in strong, specific interactions that are important in their folding and oligomerization. Crystallographic studies of 21 such helices have indicated that these interactions are similar to those described for soluble proteins. Helix-helix interactions are also important in the oligomerization of a number of proteins that have a single transmembrane α-helix. The interactions are rather specific, involving interhelical salt bridges, hydrogen bonds or precise packing interactions. In some cases, such oligomerization is required for exit from the endoplasmic reticulum. The transmembrane helices of some Golgi-residing proteins also contain sufficient information to ensure their retention in this compartment. Finally, interactions between transmembrane α-helices may be important in the mechanism of transmembrane signalling by a number of membrane-bound receptors.


title: The Origin and Evolution of Porcine Reproductive and Respiratory Syndrome Viruses

text: Porcine reproductive and respiratory syndrome viruses (PRRSV) are divided into North American and European types, which show about 40% difference in their amino acid sequences. The divergence time of these two types has been estimated to be about 1980 from epidemiological data. This suggested that PRRSV have evolved at a higher evolutionary rate (order of 10(−2)/site/year) compared with other RNA viruses of 10(−3) to 10(−5)/site/year. Here, to test the evolutionary history of PRRSV speculated by the epidemiological background, we estimated the divergence time and evolutionary rate of PRRSV with molecular evolutionary analysis. Estimated divergence time (1972–1988) corresponded well to that estimated by the epidemiological data, and the evolutionary rate (4.71–9.8) × 10(−2) of PRRSV was indeed the highest among RNA viruses so far reported. Furthermore, we inferred important sites for the adaptation in order to examine how PRRSV have adapted to swine since they emerged. The adaptive sites were located not only in the epitopes related to immunity but also in the transmembrane regions including a signal peptide. In particular, the adaptive sites in the transmembrane regions were considered to affect compatibility to the host cell membrane. We conclude that PRRSV were transmitted from another host species to swine in about 1980 and have adapted to swine by altering the transmembrane regions.


NMR spectroscopy of basic/aromatic amino acid clusters in membrane proteins


title: Real-time monitoring the transmission potential of COVID-19 in Singapore, March 2020

text: BACKGROUND: As of March 31, 2020 the ongoing COVID-19 epidemic that started in China in December 2019 is now generating local transmission around the world. The geographic heterogeneity and associated intervention strategies highlight the need to monitor in real time the transmission potential of COVID-19. Singapore provides a unique case example for monitoring transmission, as there have been multiple disease clusters, yet transmission remains relatively continued. METHODS: Here we estimate the effective reproduction number, R(t), of COVID-19 in Singapore from the publicly available daily case series of imported and autochthonous cases by date of symptoms onset, after adjusting the local cases for reporting delays as of March 17, 2020. We also derive the reproduction number from the distribution of cluster sizes using a branching process analysis that accounts for truncation of case counts. RESULTS: The local incidence curve displays sub-exponential growth dynamics, with the reproduction number following a declining trend and reaching an estimate at 0.7 (95% CI: 0.3, 1.0) during the first transmission wave by February 14, 2020 while the overall R based on the cluster size distribution as of March 17, 2020 was estimated at 0.6 (95% CI: 0.4, 1.02). The overall mean reporting delay was estimated at 6.4 days (95% CI: 5.8, 6.9), but it was shorter among imported cases compared to local cases (mean 4.3 vs. 7.6 days, Wilcoxon test, p<0.001). CONCLUSION: The trajectory of the reproduction number in Singapore underscores the significant effects of successful containment efforts in Singapore, but it also suggests the need to sustain social distancing and active case finding efforts to stomp out all active chains of transmission.


title: Classification of the cutaneous manifestations of COVID‐19: a rapid prospective nationwide consensus study in Spain with 375 cases

text: BACKGROUND: Cutaneous manifestations of COVID‐19 disease are poorly characterized. OBJECTIVES: To describe the cutaneous manifestations of COVID‐19 disease and to relate them to other clinical findings METHODS: Nationwide case collection survey of images and clinical data. Using a consensus, we described 5 clinical patterns. We later described the association of these patterns with patient demographics, timing in relation to symptoms of the disease, severity, and prognosis. RESULTS: Lesions may be classified as acral areas of erythema with vesicles or pustules (Pseudo‐chilblain) (19%), other vesicular eruptions (9%), urticarial lesions (19%), maculopapular eruptions (47%) and livedo or necrosis (6%). Vesicular eruptions appear early in the course of the disease (15% before other symptoms). The pseudo‐chilblain pattern frequently appears late in the evolution of the COVID‐19 disease (59% after other symptoms), while the rest tend to appear with other symptoms of COVID‐19. Severity of COVID‐19 shows a gradient from less severe disease in acral lesions to most severe in the latter groups. Results are similar for confirmed and suspected cases, both in terms of clinical and epidemiological findings. Alternative diagnoses are discussed but seem unlikely for the most specific patterns (pseudo‐chilblain and vesicular). CONCLUSIONS: We provide a description of the cutaneous manifestations associated with COVID‐19 infection. These may help clinicians approach patients with the disease and recognize paucisymptomatic cases.


title: Global Health Law: International Law and Public Health Policy

text: Abstract The field of global health law and the contribution of international organizations to its development are described. The growth and elaboration of the field of global health law in the last two decades is one of the most notable developments in global health policy. In this new era of global health governance, international law has an important, albeit limited, role to play in promoting and coordinating international cooperation and national action to protect and promote global health. Particular attention is paid to the global impact of binding international agreements and other international legal instruments developed under the auspices of the World Health Organization and the World Trade Organization.


title: Post-disaster evacuation and temporary resettlement considering panic and panic spread

text: Abstract After a disaster, a huge number of homeless victims should be evacuated to temporary resettlement sites. However, because the number of temporary shelters is insufficient, as are shelter building capabilities, victims must be evacuated and resettled in batches. The perceived psychological penalty to victims may increase due to heightened panic when waiting for evacuation and resettlement, whereas psychological interventions can decrease the magnitude of this panic. Based on the susceptible–infective-removal model, panic spread among homeless victims and other disaster-affected people is modeled, while considering the effects of psychological interventions on panic spread. A function is derived to compute the increase in the number of victims to be evacuated due to panic spread. A novel mixed-integer linear program is constructed for multi-step evacuation and temporary resettlement under minimization of panic-induced psychological penalty cost, psychological intervention cost, and costs associated with transportation and building shelters. The model is solved by aggregating objectives into a single objective by assigning weights to these objectives. With Wenchuan County as the test case, the epicenter of the 2008 Sichuan earthquake, the influence and the sensitivity of parameters, tradeoff among costs, and the effects of various functions of panic strength on psychological penalty and monetary costs are assessed using six experimental scenarios. Analytical results reveal the complexity and managerial insights gained by applying the proposed method to post-disaster evacuation and temporary resettlement.


title: Armed oncolytic viruses: A kick-start for anti-tumor immunity

text: Abstract Oncolytic viruses (OVs), viruses that specifically result in killing tumor cells, represent a promising class of cancer therapy. Recently, the focus in the OV therapy field has shifted from their direct oncolytic effect to their immune stimulatory effect. OV therapy can function as a “kick start” for the antitumor immune response by releasing tumor associated antigens and release of inflammatory signals. Combining OVs with immune modulators could enhance the efficacy of both immune and OV therapies. Additionally, genetic engineering of OVs allows local expression of immune therapeutics, thereby reducing related toxicities. Different options to modify the tumor microenvironment in combination with OV therapy have been explored. The possibilities and obstacles of these combinations will be discussed in this review.


Cambios de TC de tórax antes y después del tratamiento en un paciente con COVID-19[[[en]]]Pre- and Posttreatment Chest CT Changes in a patient with COVID-19


title: Cashing in on innovation: a taxonomy of FinTech

text: In this article, we provide a taxonomy of the different types of innovation occurring in financial services, a framework which we refer to as our FinTech Ecosystem. We use this FinTech Ecosystem to highlight the various areas of potential value for investors who are looking into ventures in this space as alternative investments. Our conceptual framework enables investors to examine emerging technologies and FinTech verticals through the lens of a sector analysis. To better understand where the FinTech Revolution started and where it is going, we apply the concept of digital transformation to the financial services industry which allows us to highlight the delicate risk-reward balance in FinTech.


title: Potential airborne transmission between two isolation cubicles through a shared anteroom

text: Abstract Full-scale experiments and CFD simulations were performed to study potential inter-cubicle airborne transmissions through a shared anteroom due to the hinged door opening. When doors are closed, current negative pressure designs are effective for the containment of airborne pathogens in the 'dirty' cubicle with an index patient. When the 'dirty' cubicle door is open, airborne agents can move into the other 'clean' cubicle via the shared anteroom. As the door being opened or closed, the door sweeping effect is the main source of the two-way airflow and contaminant exchange through the doorway. When the dirty cubicle door remains fully open, temperature difference and concentration gradient across the doorway induce the two-way buoyancy-driven flow and transport of airborne agents across the doorway. The longer the dirty cubicle door remains fully open (10 s, 30 s or 60 s) or the smaller the air change rate (34–8.5 ACH for each cubicle), the more airborne pathogens are being transported into the 'clean' cubicle and the longer time it takes to remove them after the door is closed. Keeping the door completely open is potentially responsible for the majority of inter-cubicle transmissions if its duration is much longer than the duration of door motion (only 3 s). Our analyses suggest a potential inter-cubicle infection risk if the shared anteroom is used for multiple isolation cubicles. Decreasing the duration of door opening, raising air change rate or using a curtain at the doorway are recommended to reduce inter-cubicle exposure hazards.


title: Distributions of HLA-A and -B alleles and haplotypes in the Yi ethnic minority of Yunnan, China: relationship to other populations

text: OBJECTIVE: To investigate the distributions of human leukocyte antigen (HLA)-A and -B alleles and HLA-A-B haplotypes in the Yi ethnic minority of the Yunnan Province, situated in southwestern China. METHODS: DNA typing for HLA-A and -B loci was performed using the polymerase chain reaction-sequence-based typing (PCR-SBT) method on 114 randomly selected healthy individuals of the Yi population. The allelic frequencies of HLA-A and -B loci were calculated by direct counting and HLA-A-B haplotypes were estimated using the expectation maximization algorithm. RESULTS: A total of 17 HLA-A and 38 HLA-B alleles were found in the Yi population. The most frequent alleles were A*2402 (32.46%), A*1101 (26.32%), and A*0203 (10.09%) at the HLA-A locus and B*4601 (12.28%), B*1525 (10.09%), B*4001 (8.77%), and B*3802 (7.89%) at the HLA-B locus. The predominant HLA-A-B haplotypes were A*2402-B*1525 (7.86%) and A*0203-B*3802 (5.64%), followed by A*1101-B*4001 (4.69%). Phylogenetic analysis indicates that the Yi population in the Honghe, Yunnan Province of China basically belongs to groups of southeastern Asian origin, but shares some characteristics with northeastern Asian groups. CONCLUSION: The present study may add to the understanding of HLA polymorphism in the Yi ethnic group that was poorly defined previously, and provide useful information for bone marrow transplantation, anthropological research, and forensic sciences as well as for disease-association studies.


title: The 2009 pandemic influenza A(H1N1) coincides with changes in the epidemiology of other viral pathogens causing acute respiratory tract infections in children

text: BACKGROUND: In Germany, the outbreak of the novel pandemic 2009 influenza A(H1N1) virus A(H1N1)pdm09 caused a wave of high activity between November 2009 and January 2011. The aim of this study was to investigate the prevalence of 19 respiratory pathogens in children hospitalized for lower respiratory tract infections during the winter influenza seasons of 2009/2010 and 2010/2011 and to observe a possible impact of influenza A(H1N1)pdm09 on the epidemiology of other epidemic viruses. MATERIALS AND METHODS: Specimens were nasopharyngeal aspirates which had been collected from children admitted to the participating hospitals in the area of Mainz, Wiesbaden, and Kiel, Germany, with acute community-acquired lower respiratory tract infections. The specimens were subjected to a previously described multiplex reverse transcription PCR assay to detect the following microorganisms: enterovirus, influenza virus types A and B, respiratory syncytial virus (RSV), parainfluenzavirus types 1–4, adenovirus, Mycoplasma pneumoniae, Chlamydophila pneumoniae, rhinovirus, human metapneumovirus (hMPV), coronavirus OC43 and 229E, influenza A(H1N1)pdm09, Bordetella pertussis, Bordetella parapertussis, and Legionella pneumophila. RESULTS: A total of 3,998 clinical specimens were collected from July 2009 to March 2011, of which 296 were positive for A(H1N1)pdm09. An epidemic of seasonal influenza A or B was not observed in the 2009/2010 season, but a minor epidemic of seasonal influenza B was observed in January/February 2011. Influenza A(H1N1)pdm09 coincided with the absence of the seasonal influenza A of former years. The RSV and hMPV epidemics of 2009/2010 erupted several weeks later than expected based on data collected in the PID-ARI-Network during the past 10 years, whereas in the 2010/2011 influenza season they occurred as expected. CONCLUSIONS: The emergence of the novel influenza A(H1N1)pdm09 virus may have been influenced the epidemiology of other epidemic viruses, such as the RSV and hMPV. No epidemic of seasonal influenza was observed in the 2009/2010 influenza season.


title: Pulmonary presentation of Kawasaki disease—A diagnostic challenge

text: OBJECTIVES: Kawasaki disease (KD) is a multisystemic vasculitis with predominant mucocutaneous manifestations. Pulmonary involvement in KD is distinctly uncommon and is not commonly recognized. We describe our experience of managing children with KD wherein the initial presentation was predominantly pulmonary. METHODS: Six hundred and two children have been diagnosed with KD during the period January 1993 to May 2017 in the Allergy Immunology Unit, Advanced Pediatrics Centre, Postgraduate Institute of Medical Education and Research, Chandigarh. Data were collected from inpatient records in Allergy Immunology Unit and follow‐up files in the Pediatric Rheumatology Clinic. RESULTS: Of 602 children, 11 (1.83%) had a predominant pulmonary presentation of KD. Mean age at diagnosis of KD was 2.5 years. Fever, cough and respiratory distress were the presenting complaints in all patients. First sign of KD was noted at a mean duration of 14.5 days from the onset of symptoms. Periungual desquamation was the most common clinical sign (72.7%). Persistent fever in spite of antimicrobials, thrombocytosis, and elevated erythrocyte sedimentation rate and C‐reactive protein levels pointed toward a diagnosis of KD in our patients. Parenchymal consolidation was evident on chest X‐ray in all patients, pleural effusion in six, empyema in three, and pneumothorax in two patients. Coronary artery abnormalities were evident in three patients. Intravenous immunoglobulin was given after a mean period of 22.4 days of onset of fever. CONCLUSIONS: The diagnosis of KD is often delayed in children who have a predominantly pulmonary presentation. This can have adverse clinical consequences.


title: Macrophages: Their role, activation and polarization in pulmonary diseases

text: Abstract Macrophages, circulating in the blood or concatenated into different organs and tissues constitute the first barrier against any disease. They are foremost controllers of both innate and acquired immunity, healthy tissue homeostasis, vasculogenesis and congenital metabolism. Two hallmarks of macrophages are diversity and plasticity due to which they acquire a wobbling array of phenotypes. These phenotypes are appropriately synchronized responses to a variety of different stimuli from either the tissue microenvironment or – microbes or their products. Based on the phenotype, macrophages are classified into classically activated/(M1) and alternatively activated/(M2) which are further sub-categorized into M2a, M2b, M2c and M2d based upon gene expression profiles. Macrophage phenotype metamorphosis is the regulating factor in initiation, progression, and termination of numerous inflammatory diseases. Several transcriptional factors and other factors controlling gene expression such as miRNAs contribute to the transformation of macrophages at different points in different diseases. Understanding the mechanisms of macrophage polarization and modulation of their phenotypes to adjust to the micro environmental conditions might provide us a great prospective for designing novel therapeutic strategy. In view of the above, this review summarises the activation of macrophages, the factors intricated in activation along with benefaction of macrophage polarization in response to microbial infections, pulmonary toxicity, lung injury and other inflammatory diseases such as chronic obstructive pulmonary dysplasia (COPD), bronchopulmonary dysplasia (BPD), asthma and sepsis, along with the existing efforts to develop therapies targeting this facet of macrophage biology.


title: Clinically significant anticardiolipin antibodies associated with COVID-19()

text: The novel coronavirus strain known as SARS-CoV-2 has rapidly spread around the world creating distinct challenges to the healthcare workforce. Coagulopathy contributing to significant morbidity in critically ill patients with SARS-CoV-2 has now been well documented. We discuss two cases selected from patients requiring critical care in April 2020 in New York City with a unique clinical course. Both cases reveal significant thrombotic events noted on imaging during their hospital course. Obtaining serial inflammatory markers in conjunction with anti-phospholipid antibody testing revealed clinically significant Antiphospholipid syndrome (APS). This case series reviews the details preceding APS observed in SARS-CoV-2 and aims to report findings that could potentially further our understanding of the disease.


title: Downstream Processing: From Egg to Cell Culture‐Derived Influenza Virus Particles

text: The establishment of cell culture‐derived vaccine production requires the development of appropriate downstream processes. Until today, many of the downstream methods applied originate from egg‐derived production processes. These methods have often been slightly modified in order to account for the new demands. However, efforts are currently underway to optimize these processes focusing, for example, on ion exchange or affinity based membrane adsorption chromatography. This review covers the main aspects relevant for the downstream processing of egg and mammalian cell culture‐derived whole influenza viruses.


title: Two predominant MUPs, OBP3 and MUP13, are male pheromones in rats

text: BACKGROUND: In rats, urine-borne male pheromones comprise organic volatile compounds and major urinary proteins (MUPs). A number of volatile pheromones have been reported, but no MUP pheromones have been identified in rat urine. RESULTS: We used sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing electrophoresis (IEF), nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) after in gel digestion of the proteins and quantitative real-time PCR (qRT-PCR) and showed that the levels of two MUPs, odorant-binding protein 3 (OBP3) (i.e. PGCL4) and MUP13 (i.e. PGCL1), in urine and their mRNAs in liver were higher in males than in females and were suppressed by orchidectomy and restored by testosterone treatment (T treatment). We then generated recombinant MUPs (rMUPs) and found that the sexual attractiveness of urine from castrated males to females significantly increased after the addition of either recombinant OBP3 (rOBP3) or recombinant MUP13 (rMUP13). Using c-Fos immunohistochemistry, we further examined neuronal activation in the brains of female rats after they sniffed rOBP3 or rMUP13. Both rOBP3 and rMUP13 activated the accessory olfactory bulb (AOB), medial preoptic area (MPA), bed nucleus of the stria terminalis (BST), medial amygdala (MeA), posteromedial cortical amygdala (PMCo) and ventromedial nucleus of the hypothalamus (VMH), which participate in the neural circuits responsible for pheromone-induced sexual behaviours. In particular, more c-Fos-immunopositive (c-Fos-ir) cells were observed in the posterior AOB than in the anterior AOB. CONCLUSIONS: The expression of OBP3 and MUP13 was male-biased and androgen-dependent. They attracted females and activated brain areas related to sexual behaviours in female rats, suggesting that both OBP3 and MUP13 are male pheromones in rats. Particularly, an OBP excreted into urine was exemplified to be a chemical signal.


title: Characteristics of personal protective equipment training programs in Australia and New Zealand hospitals: A survey

text: BACKGROUND: Using personal protective equipment (PPE) is one of several fundamental measures to prevent the transmission of infection and infectious diseases and is particularly pertinent in the current COVID-19 pandemic. Appropriate use of PPE by healthcare workers is, however, often suboptimal. Training and monitoring of PPE competency are essential components of an infection prevention and control program but there is a paucity of research and data on the content of such training programs across Australasia. This paper reports the results of a survey that characterised the nature of PPE training in Australian and New Zealand hospitals. METHODS: A population-based online survey was distributed to members of three major Australasian colleges representing infection prevention and control. RESULTS: Results indicate that, although training is frequently provided at orientation, many healthcare workers do not receive regular updates. Training programmes combine online and classroom sessions, but over a third do not include a practical component. The frequency of monitoring PPE competency is variable with one third of respondents indicating that no auditing occurs. PPE items used for high-level training are variable, with use of powered air purifying respirators (PAPRs) uncommon. CONCLUSION: The results of this study suggest that HCWs’ confidence, competence and familiarity with PPE are a concern, which in the context of the current global COVID-19 pandemic is problematic. More research is needed into how PPE training programs could be better designed, to prepare HCWs for practice using PPE safely and confidently.


Mental health status of health care workers during the COVID‐19 outbreak in Poland: one region, two different settings


title: Recent advances in nanowires-based field-effect transistors for biological sensor applications

text: Abstract Nanowires (NWs)-based field-effect transistors (FETs) have attracted considerable interest to develop innovative biosensors using NWs of different materials (i.e. semiconductors, polymers, etc.). NWs-based FETs provide significant advantages over the other bulk or non-NWs nanomaterials-based FETs. As the building blocks for FET-based biosensors, one-dimensional NWs offer excellent surface-to-volume ratio and are more suitable and sensitive for sensing applications. During the past decade, FET-based biosensors are smartly designed and used due to their great specificity, sensitivity, and high selectivity. Additionally, they have the advantage of low weight, low cost of mass production, small size and compatible with commercial planar processes for large-scale circuitry. In this respect, we summarize the recent advances of NWs-based FET biosensors for different biomolecule detection i.e. glucose, cholesterol, uric acid, urea, hormone, proteins, nucleotide, biomarkers, etc. A comparative sensing performance, present challenges, and future prospects of NWs-based FET biosensors are discussed in detail.


title: Porcine IL-12 plasmid as an adjuvant improves the cellular and humoral immune responses of DNA vaccine targeting transmissible gastroenteritis virus spike gene in a mouse model

text: Transmissible gastroenteritis (TGE), caused by transmissible gastroenteritis virus (TGEV), is a highly infectious disease in pigs. Vaccination is an effective approach to prevent TGEV infection. Here, we evaluated the potential of TGEV S1 as a DNA vaccine and porcine interleukin (pIL)-12 as an adjuvant in a mouse model. A DNA vaccine was constructed with the TGEV S1 gene to induce immune response in an experimental mouse model; pIL-12 was chosen as the immunological adjuvant within this DNA vaccine. The pVAX1-(TGEV-S1) and pVAX1-(pIL-12) vectors were transfected into BHK-21 cells and expressed in vitro. Experimental mice were separately immunized with each of the recombinant plasmids and controls through the intramuscular route. The lymphocytes isolated from the blood and spleen were analyzed for proliferation, cytotoxic activities, and populations of CD4(+) and CD8(+) cells. The titers of TGEV S1 in an enzyme-linked immunosorbent assay (ELISA) and TGEV neutralizing antibodies and the concentrations of interferon (IFN)-γ and IL-4 were also analyzed in the serum. The plasmids pVAX1-(TGEV-S1) and pVAX1-(pIL-12) could be expressed in BHK-21 cells, and the combination of pVAX1-(TGEV-S1) and pVAX1-(pIL-12) could induce a significant increase in all markers. pIL-12 could act as an immunological adjuvant in the DNA vaccine for TGEV-S1. Furthermore, the DNA vaccine prepared using TGEV-S1 and porcine IL-12 could induce excellent humoral and cellular immune responses.


title: Antihyperglycemic properties of hydroxychloroquine in patients with diabetes: risks and benefits at the time of COVID‐19 pandemic

text: The antimalarial drug hydroxychloroquine (HCQ) has long been used as a disease‐modifying antirheumatic drug for the treatment of several inflammatory rheumatic diseases. Over the last three decades, various studies have shown that HCQ plays also a role in the regulation of glucose homeostasis. Although the mechanisms of action underlying the glucose‐lowering properties of HCQ are still not entirely clear, evidence suggests that this drug may exert multifaceted effects on glucose regulation, including improvement of insulin sensitivity, increase of insulin secretion, reduction of hepatic insulin clearance and reduction of systemic inflammation. Preliminary studies have shown the safety and efficacy of HCQ (at a dose ranging from 400 to 600 mg/day) in patients with type 2 diabetes over a short‐term period. In 2014, HCQ has been approved in India as an add‐on hypoglycemic agent for patients with uncontrolled type 2 diabetes. However, large randomized controlled trials are needed to establish the safety and efficacy profile of HCQ in patients with type 2 diabetes over a long‐term period. With regard to the COVID‐19 pandemic, several medications (including HCQ) have been used as off‐label drugs due to the lack of proven effective therapies. However, emerging evidence shows limited benefit from HCQ use in COVID‐19 in general. The aim of this manuscript is to comprehensively summarize the current knowledge on the antihyperglycemic properties of HCQ and to critically evaluate the potential risks and benefits related to HCQ use in patients with diabetes, even in light of the current pandemic scenario. This article is protected by copyright. All rights reserved.


title: A Novel Indicator Predicts 2019 Novel Coronavirus Infection in Subjects with Diabetes

text: Abstract Aims Diabetes mellitus (DM) is associated with significant morbidity and mortality. The disease severity in 2019 novel coronavirus (Covid 19) infection has varied from mild self-limiting flu-like illness to fulminant pneumonia, respiratory failure and death. Since DM and Covid 19 infection are closely associated with inflammatory status, mean platelet volume (MPV) was suggested to be useful in predicting Covid infection onset. This study aimed to evaluate the diagnostic role of MPV in Covid patients with diabetes. Methods A total of 640 subjects (160 Covid patients with type 2 diabetes, 160 healthy controls, 160 patients with non-spesific infections and 160 Covid patients without type 2 diabetes) enrolled in the study. Results MPV was significantly higher (11.21 ± 0.61 fL) as compared to the results from the last routine visits of the the same individuals with diabetes (10.59 ± 0.96 fL) (p = 0.000). Conclusions MPV could be used as a simple and cost-effective tool to predict the Covid infection in subjects with diabetes in primary care.


title: Azithromycin added to hydroxychloroquine for patients admitted to intensive care due to coronavirus disease 2019 (COVID-19)—protocol of randomised controlled trial AZIQUINE-ICU

text: BACKGROUND: Novel coronavirus SARS-CoV-2 is known to be susceptible in vitro to exposure to hydroxychloroquine and its effect has been found to be potentiated by azithromycin. We hypothesise that early administration of hydroxychloroquine alone or in combination with azithromycin can prevent respiratory deterioration in patients admitted to intensive care due to rapidly progressive COVID-19 infection. METHODS: Design: Prospective, multi-centre, double-blind, randomised, controlled trial (RCT). Participants: Adult (> 18 years) within 24 h of admission to the intensive care unit with proven or suspected COVID-19 infection, whether or not mechanically ventilated. Exclusion criteria include duration symptoms of febrile disease for ≥ 1 week, treatment limitations in place or moribund patients, allergy or intolerance of any study treatment, and pregnancy. Interventions: Patients will be randomised in 1:1:1 ratio to receive Hydroxychloroquine 800 mg orally in two doses followed by 400 mg daily in two doses and azithromycin 500 mg orally in one dose followed by 250 mg in one dose for a total of 5 days (HC-A group) or hydroxychloroquine + placebo (HC group) or placebo + placebo (C-group) in addition to the best standard of care, which may evolve during the trial period but will not differ between groups. Primary outcome is the composite percentage of patients alive and not on end-of-life pathway who are free of mechanical ventilation at day 14. Secondary outcomes: The percentage of patients who were prevented from needing intubation until day 14, ICU length of stay, and mortality (in hospital) at day 28 and 90. DISCUSSION: Although both investigational drugs are often administered off label to patients with severe COVID-19, at present, there is no data from RCTs on their safety and efficacy. In vitro and observational trial suggests their potential to limit viral replication and the damage to lungs as the most common reason for ICU admission. Therefore, patients most likely to benefit from the treatment are those with severe but early disease. This trial is designed and powered to investigate whether the treatment in this cohort of patients leads to improved clinical patient-centred outcomes, such as mechanical ventilation-free survival. TRIAL REGISTRATION: Clinical trials.gov: NCT04339816 (Registered on 9 April 2020, amended on 22 June 2020); Eudra CT number: 2020-001456-18 (Registered on 29 March 2020).


title: Genomic characterization of circoviruses associated with acute gastroenteritis in minks in northeastern China

text: Mink circovirus (MiCV), a virus that was newly discovered in 2013, has been associated with enteric disease. However, its etiological role in acute gastroenteritis is unclear, and its genetic characteristics are poorly described. In this study, the role of circoviruses (CVs) in mink acute gastroenteritis was investigated, and the MiCV genome was molecularly characterized through sequence analysis. Detection results demonstrated that MiCV was the only pathogen found in this infection. MiCVs and previously characterized CVs shared genome organizational features, including the presence of (i) a potential stem-loop/nonanucleotide motif that is considered to be the origin of virus DNA replication; (ii) two major inversely arranged open reading frames encoding putative replication-associated proteins (Rep) and a capsid protein; (iii) direct and inverse repeated sequences within the putative 5ʹ region; and (iv) motifs in Rep. Pairwise comparisons showed that the capsid proteins of MiCV shared the highest amino acid sequence identity with those of porcine CV (PCV) 2 (45.4%) and bat CV (BatCV) 1 (45.4%). The amino acid sequence identity levels of Rep shared by MiCV with BatCV 1 (79.7%) and dog CV (dogCV) (54.5%) were broadly similar to those with starling CV (51.1%) and PCVs (46.5%). Phylogenetic analysis indicated that MiCVs were more closely related to mammalian CVs, such as BatCV, PCV, and dogCV, than to other animal CVs. Among mammalian CVs, MiCV and BatCV 1 were the most closely related. This study could contribute to understanding the potential pathogenicity of MiCV and the evolutionary and pathogenic characteristics of mammalian CVs.


title: Angiotensin‐converting enzyme 2: The old door for new severe acute respiratory syndrome coronavirus 2 infection

text: Coronavirus (CoV) disease 2019 (COVID‐19) is an ongoing pandemic caused by severe acute respiratory syndrome CoV 2 (SARS‐CoV‐2). The highly contagious SARS‐CoV‐2 belongs to the genus Betacoronavirus, and it is phylogenetically closely related to SARS‐CoV, a human CoV that caused an outbreak back in 2002 to 2003. Both SARS‐CoV‐2 and SARS‐CoV enter human cells via the interactions between viral crown‐like spike protein and human angiotensin‐converting enzyme 2 (ACE2) receptor. Here, we aim to review the involvement of ACE2 in human CoV infections by discussing the roles of ACE2 in CoV evolution, cross‐species transmissibility, and COVID‐19 susceptibility. We also provide our perspectives on COVID‐19 treatment and prevention.


title: Immunogenicity and protection efficacy of monovalent and polyvalent poxvirus vaccines that include the D8 antigen

text: Abstract Recent studies have established the feasibility of subunit-based experimental vaccines to protect animals from lethal poxvirus infection. Individual outer membrane proteins from intracellular and extracellular virions of vaccinia virus, when delivered in the form of either DNA vaccines or recombinant protein vaccines produced from baculovirus-infected insect cells, were able to protect mice from the vaccinia virus challenge and rhesus macaques from the monkeypox virus challenge. The polyvalent formulations with various combinations of the four poxvirus antigens (A27, L1, B5 and A33) achieved better protection than the monovalent formulation using only one of these antigens. However, it is not clear whether any of the remaining outer membrane poxvirus proteins can further improve the efficacy of the current polyvalent formulations. In this study, we conducted detailed analysis on the immunogenicity of D8, a previously reported protective antigen from intracellular mature virions. Our results indicated that D8 induced strong protective antibody responses and was effective in improving the efficacy of previously reported polyvalent poxvirus vaccine formulations. Therefore, D8 is an excellent candidate antigen to be included in the final polyvalent subunit-based poxvirus vaccines.


title: Endogenous opiates: 1993

text: Abstract This paper is the sixteenth installment of our annual review of research concerning the opiate system. It is restricted to papers published during 1993 that concern the behavioral effects of the endogenous opiate peptides, and does not include papers dealing only with their analgesic properties. The specific topics this year include stress; tolerance and dependence; eating; drinking; gastrointestinal, renal, and hepatic function; mental illness and mood; learning, memory, and reward; cardiovascular responses; respiration and thermoregulation; seizures and other neurological disorders; electrical-related activity; general activity and locomotion; development; immunological responses; and other behaviors.


Virus Etiology of Airway Illness in Elderly Adults


title: Seasonal variation in the incidence of necrotizing enterocolitis

text: BACKGROUND: Necrotizing enterocolitis (NEC) has been reported to occur in a sporadic manner and in clusters of cases. We hypothesized that variations in the incidence of NEC were random, without clustering. In order to define the pattern of NEC in the United States, we analyzed the Pediatric Health Information System (PHIS) database to evaluate whether NEC cases are distributed randomly or exhibit temporal clustering or periodicity. METHODS: After obtaining IRB approval, we queried the PHIS database for all patients with NEC (ICD-9 code of 777.5) over a 13-year period. Sixteen children’s hospitals were studied, and individual institutional and aggregate data were reviewed. Fisher’s Kappa and Bartlett’s Kolmogorov–Smirnov tests were used to identify periodicity. RESULTS: During the study interval, there were 2,93,076 neonatal admissions, and 4,559 (1.6%) infants were diagnosed with NEC. Statistical analysis demonstrates a periodicity of 6 months in the occurrence of NEC. Fisher’s Kappa was 16.924 and Bartlett’s Kolmogorov–Smirnov was 0.281, which translates to a P value of <0.0001. CONCLUSIONS: On the basis of a national database analysis it appears a temporally non-random distribution of NEC cases does exist.


title: Symmetric cutaneous vasculitis in COVID‐19 pneumonia

text: Fever, cough, breathing difficulties, digestive issues and loss of smell and taste are the most common symptoms of novel SARS-CoV2 infection but cutaneous manifestations have been highlighted by several dermatologists. We found reports (1-3) to be very interesting because it was underlined how the COVID19 infection can also give cutaneous manifestations vasculitis - like. It has already been described how purple in children, when accompanied by fever, can be a rare but possible manifestation of novel SARs-CoV2 infection(4).


Virtual Interviews for the Complex General Surgical Oncology Fellowship: The Dana-Farber/Partners Experience


title: The natural compound silvestrol inhibits hepatitis E virus (HEV) replication in vitro and in vivo

text: Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and a member of the genus Orthohepevirus in the family Hepeviridae. HEV infections are the common cause of acute hepatitis but can also take chronic courses. Ribavirin is the treatment of choice for most patients and type I interferon (IFN) has been evaluated in a few infected transplantation patients in vivo. However, no effective and specific treatments against HEV infections are currently available. In this study, we evaluated the natural compound silvestrol, isolated from the plant Aglaia foveolata, and known for its specific inhibition of the DEAD-box RNA helicase eIF4A in state-of-the-art HEV experimental model systems. Silvestrol blocked HEV replication of different subgenomic replicons in a dose-dependent manner at low nanomolar concentrations and acted additive to ribavirin (RBV). In addition, HEV p6-based full length replication and production of infectious particles was reduced in the presence of silvestrol. A pangenotypic effect of the compound was further demonstrated with primary isolates from four different human genotypes in HEV infection experiments of hepatocyte-like cells derived from human embryonic and induced pluripotent stem cells. In vivo, HEV RNA levels rapidly declined in the feces of treated mice while no effect was observed in the vehicle treated control animals. In conclusion, silvestrol could be identified as pangenotypic HEV replication inhibitor in vitro with additive effect to RBV and further demonstrated high potency in vivo. The compound therefore may be considered in future treatment strategies of chronic hepatitis E in immunocompromised patients.


title: Ventilators, Guidelines, Judgment, and Trust

text: Covid‐19 confronts us with tragic choices, in which every option is unacceptable. On the New York State Task Force on Life and the Law, I worked on guidelines for such situations. We did not envision the scale or character of Covid‐19. To minimize fear that the decisions made in these situations might be unfair, we all must know what guidelines or mandates inform them. Only with transparency about how decisions will be made, by whom, and according to what requirements can we have confidence that fairness prevails. We now face many related questions about process, goals, leadership, and trust. For example, how might ethical guidelines evolve as scientific understanding advances? Should guidelines vary with different venues? And if, as I have argued, judgment is necessary even with the best of guidelines, how can we prepare clinicians to make good judgments? Are there implications for better training of personnel in nonemergency times for what they might face in the worst of times?


Principles of ethical prescribing for self and others: hydroxychloroquine in the COVID-19 pandemic


title: Impact of norepinephrine on right ventricular afterload and function in septic shock—a strain echocardiography study

text: BACKGROUND: In this observational study, the effects of norepinephrine‐induced changes in mean arterial pressure (MAP) on right ventricular (RV) systolic function, afterload and pulmonary haemodynamics were studied in septic shock patients. We hypothesised that RV systolic function improves at higher doses of norepinephrine/MAP levels. METHODS: Eleven patients with septic shock requiring norepinephrine after fluid resuscitation were included <24 hours after ICU arrival. Study enrolment and insertion of a pulmonary artery catheter was performed after written informed consent from the next of kin. Norepinephrine infusion was titrated to target mean arterial pressures (MAP) of 60, 75 and 90 mmHg in a random sequential order. At each target MAP, strain—and conventional echocardiographic—and pulmonary haemodynamic variables were measured. RV afterload was assessed as effective pulmonary arterial elastance, (E (pa)) and pulmonary vascular resistance index, (PVRI). RV free wall peak strain was the primary end‐point. RESULTS: At highest compared to lowest norepinephrine dose/MAP level, RV free wall peak strain increased from −19% to −25% (32%, P = .003), accompanied by increased tricuspid annular plane systolic excursion (22%, P = .01). At the highest norepinephrine dose/MAP, RV end‐diastolic area index (16%, P < .001), central venous pressure (38%, P < .001), stroke volume index (7%, P = .001), mean pulmonary artery pressure (19%, P < .001) and RV stroke work index (15%, P = .045) increased, with no effects on PVRI or E (pa). Cardiac index did not change, assessed by thermodilution (P = .079) and echocardiography (P = .054). CONCLUSIONS: Higher doses of norepinephrine to a target MAP of 90 mm Hg improved RV systolic function while RV afterload was not affected.


title: RNA synthetic mechanisms employed by diverse families of RNA viruses

text: RNA viruses are ubiquitous in nature, infecting every known organism on the planet. These viruses can also be notorious human pathogens with significant medical and economic burdens. Central to the lifecycle of an RNA virus is the synthesis of new RNA molecules, a process that is mediated by specialized virally encoded enzymes called RNA‐dependent RNA polymerases (RdRps). RdRps directly catalyze phosphodiester bond formation between nucleoside triphosphates in an RNA‐templated manner. These enzymes are strikingly conserved in their structural and functional features, even among diverse RNA viruses belonging to different families. During host cell infection, the activities of viral RdRps are often regulated by viral cofactor proteins. Cofactors can modulate the type and timing of RNA synthesis by directly engaging the RdRp and/or by indirectly affecting its capacity to recognize template RNA. High‐resolution structures of RdRps as apoenzymes, bound to RNA templates, in the midst of catalysis, and/or interacting with regulatory cofactor proteins, have dramatically increased our understanding of viral RNA synthetic mechanisms. Combined with elegant biochemical studies, such structures are providing a scientific platform for the rational design of antiviral agents aimed at preventing and treating RNA virus‐induced diseases. WIREs RNA 2013, 4:351–367. doi: 10.1002/wrna.1164 1.. RNA Interactions with Proteins and Other Molecules > RNA–Protein Complexes; 2.. RNA in Disease and Development > RNA in Disease.


Bioethics, health, and inequality


title: Prenatal anxiety and obstetric decisions among pregnant women in Wuhan and Chongqing during the COVID‐19 outbreak: a cross‐sectional study

text: OBJECTIVES: To investigate the mental status of pregnant women and to determine their obstetric decisions during the COVID‐19 outbreak. DESIGN: Cross‐sectional study. SETTING: Two cities in China, Wuhan (epicentre) and Chongqing (a less affected city). POPULATION: 1947 pregnant women. METHODS: We collected demographic, pregnancy, and epidemic information from our pregnant subjects, along with their attitudes towards COVID‐19 (using a self‐constructed five‐point scale). The Self‐Rating Anxiety Scale (SAS) was used to assess anxiety status. Obstetric decision‐making was also evaluated. The differences between cities in all of the above factors were compared, and the factors that influenced anxiety levels were identified by multivariable analysis. MAIN OUTCOME MEASURES: Anxiety status and its influencing factors. Obstetric decision‐making. RESULTS: Differences were observed between cities in some background characteristics and women's attitudes towards COVID‐19 in Wuhan were more extreme. More women in Wuhan felt anxious (24.5% vs 10.4%). Factors that influenced anxiety also included household income, subjective symptoms, and attitudes. Overall, obstetric decisions also revealed city‐based differences; these decisions mainly concerned hospital preference, time of prenatal care or delivery, mode of delivery and infant feeding. CONCLUSIONS: The outbreak aggravated prenatal anxiety, and the associated factors could be targets for psychological care. In parallel, key obstetric decision‐making changed, emphasising the need for pertinent professional advice. Special support is essential for pregnant mothers during epidemics.


title: Involvement of autophagy in viral infections: antiviral function and subversion by viruses

text: Autophagy is a cellular process involved in the degradation and turn-over of long-lived proteins and organelles, which can be subjected to suppression or further induction in response to different stimuli. According to its essential role in cellular homeostasis, autophagy has been implicated in several pathologies including cancer, neurodegeneration and myopathies. More recently, autophagy has been described as a mechanism of both innate and adaptive immunity against intracellular bacteria and viruses. In this context, autophagy has been proposed as a protective mechanism against viral infection by degrading the pathogens into autolysosomes. This is strengthened by the fact that several proteins involved in interferon (IFN) signalling pathways are linked to autophagy regulation. However, several viruses have evolved strategies to divert IFN-mediated pathways and autophagy to their own benefit. This review provides an overview of the autophagic process and its involvement in the infection by different viral pathogens and of the connections existing between autophagy and proteins involved in IFN signalling pathways.


title: Immunité innée et cognitive aux interfaces muqueuses

text: Résumé Les interfaces épithéliales muqueuses, indispensables aux relations de l'organisme avec son environnement digestif, respiratoire, reproductif, sont défendues par un système immunitaire apparaissant de plus en plus élaboré et contrôlé. Il comporte une participation efficace des cellules et des molécules de l'immunité innée, ensemble physiologique de connaissance récente, mais phylogénétiquement très ancien. Les muqueuses sont d'autre part colonisées par des cellules immunitaires (cellules présentatrices d'antigène, lignées lymphocytaires T, B), mobiles entre un site inducteur principal, les plaques de Peyer, et des sites effecteurs diversifiés et produisant des molécules effectrices et régulatrices originales (IgA sécrétoires, cytokines, chemokines). Cet ensemble complexe est physiologiquement particulièrement contrôlé et reste habituellement silencieux. Il peut néanmoins être le siège de pathologies spécifiques et commence à pouvoir être exploré biologiquement. Le progrès des connaissances fait espérer des modalités thérapeutiques nouvelles à impact muqueux. Abstract Mucosal epithelial interfaces, essential for a proper interaction of the individual with the environment of the digestive, respiratory or reproductive tracts, are protected by a powerful immune system that appears more and more sophisticated and controlled. This system involves the newly described cells and molecules of innate immunity, which appear phylogenetically highly conserved. Mucosae are moreover seeded by immunocompetent cells (antigen presenting cells, B and T lymphocytes), that traffic between the major inductive sites of Peyer's patches, and diversified effector sites. These cells elaborate original effector and regulatory molecules (secretory IgA, cytokines, chemokines). This complex and tightly controlled system is usually quite silent. Specific pathologies may however develop, the exploration of which being progressively more possible. Increasing knowledge about the mucosae associated immune system suggests that new treatments with mucosal impact should develop in the future.


Pandemics and the consequences of COVID‐19


title: Clinical predictors of COVID-19 mortality

text: BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has affected over millions of individuals and caused hundreds of thousands of deaths worldwide. It can be difficult to accurately predict mortality among COVID-19 patients presenting with a spectrum of complications, hindering the prognostication and management of the disease. METHODS: We applied machine learning techniques to clinical data from a large cohort of 5,051 COVID-19 patients treated at the Mount Sinai Health System in New York City, the global COVID-19 epicenter, to predict mortality. Predictors were designed to classify patients into Deceased or Alive mortality classes and were evaluated in terms of the area under the receiver operating characteristic (ROC) curve (AUC score). FINDINGS: Using a development cohort (n=3,841) and a systematic machine learning framework, we identified a COVID-19 mortality predictor that demonstrated high accuracy (AUC=0·91) when applied to test sets of retrospective (n= 961) and prospective (n=249) patients. This mortality predictor was based on five clinical features: age, minimum O(2) saturation during encounter, type of patient encounter (inpatient vs. various types of outpatient and telehealth encounters), hydroxychloroquine use, and maximum body temperature. INTERPRETATION: An accurate and parsimonious COVID-19 mortality predictor based on five features may have utility in clinical settings to guide the management and prognostication of patients affected by this disease.


title: The Role of Apoptin in Chicken Anemia Virus Replication

text: Apoptin is the Vp3 protein of chicken anemia virus (CAV), which infects the thymocytes and erythroblasts in young chickens, causing chicken infectious anemia and immunosuppression. Apoptin is highly studied for its ability to selectively induce apoptosis in human tumor cells and, thus, is a protein of interest in anti-tumor therapy. CAV apoptin is known to localize to different subcellular compartments in transformed and non-transformed cells, depending on the DNA damage response, and the phosphorylation of several identified threonine residues. In addition, apoptin interacts with molecular machinery such as the anaphase promoting complex/cyclosome (APC/C) to inhibit the cell cycle and induce arrest in G2/M phase. While these functions of apoptin contribute to the tumor-selective effect of the protein, they also provide an important fundamental framework to apoptin’s role in viral infection, pathogenesis, and propagation. Here, we reviewed how the regulation, localization, and functions of apoptin contribute to the viral life cycle and postulated its importance in efficient replication of CAV. A model of the molecular biology of infection is critical to informing our understanding of CAV and other related animal viruses that threaten the agricultural industry.


title: An enzyme immunoassay for identification and quantification of infectious murine parvovirus in cultured cells

text: Abstract An enzyme immunoassay was developed to identify and quantify infectious murine parvovirus. The assay was as sensitive as the fluorescent focus test for quantification of virus in clinical material and was less labor-intensive. Viral infectivity could be inactivated with formalin prior to performance of the enzyme immunoassay. Specificity of the reaction product was shown by neutralization of infectivity using a hyperimmune rabbit antiserum to murine parvovirus.


Update on infection prevention in disaster planning: New resources and policies


title: Genetic variability of human metapneumovirus amongst an all ages population in Cambodia between 2007 and 2009

text: Abstract First identified in 2001, human metapneumovirus (HMPV) is a novel pathogen and causative agent of acute respiratory tract infection. Re-infection with HMPV is common, and currently there is no available vaccine against HMPV infection. Two genotypes of HMPV have been identified, A and B, both of which can be divided further into at least two distinct sub-genotypes. Here we report the results of the first study to investigate the genetic variability of HMPV strains circulating within Cambodia. The overall incidence of HMPV infection amongst an all-ages population of patients hospitalised with ALRI in Cambodia during 3 consecutive years, between 2007 and 2009, was 1.7%. The incidence of HMPV infection was highest amongst children less than 5 years of age, with pneumonia or bronchopneumonia the most frequent clinical diagnoses across all age groups. The incidence of HMPV infection varied annually. As anticipated, genetic diversity was low amongst the conserved F gene sequences but very high amongst G gene sequences, some strains sharing as little as 56.3% and 34.2% homology at the nucleotide and amino acid levels, respectively. Simultaneous co-circulation of strains belonging to the HMPV sub-genotypes B1, B2 and lineage A2b, amongst patients recruited at 2 geographically distinct provincial hospitals, was detected. Sub-genotype B2 strains were responsible for the majority of the infections detected, and a significant (p = 0.013) association between infection with lineage A2b strains and disease severity was observed.


title: Human Coronavirus NL63 Infection in Canada

text: The isolation of human coronavirus NL63 (HCoV-NL63) in The Netherlands raised questions about its contribution to respiratory illness. In this study, a total of 525 respiratory specimens, collected in Canada primarily during the winter months of 2001–2002, were tested for HCoV-NL63; 19 tested positive for HCoV-NL63, demonstrating virus activity during January–March 2002. Patients with HCoV-NL63 were 1 month-100 years old (median age, 37 years). The main clinical presentations were fever (15/19), sore throat (5/19), and cough (9/19), and 4 patients were hospitalized. These results provide evidence for the worldwide distribution of HCoV-NL63.


title: Epidemic Surveillance Using an Electronic Medical Record: An Empiric Approach to Performance Improvement

text: BACKGROUNDS: Electronic medical records (EMR) form a rich repository of information that could benefit public health. We asked how structured and free-text narrative EMR data should be combined to improve epidemic surveillance for acute respiratory infections (ARI). METHODS: Eight previously characterized ARI case detection algorithms (CDA) were applied to historical EMR entries to create authentic time series of daily ARI case counts (background). An epidemic model simulated influenza cases (injection). From the time of the injection, cluster-detection statistics were applied daily on paired background+injection (combined) and background-only time series. This cycle was then repeated with the injection shifted to each week of the evaluation year. We computed: a) the time from injection to the first statistical alarm uniquely found in the combined dataset (Detection Delay); b) how often alarms originated in the background-only dataset (false-alarm rate, or FAR); and c) the number of cases found within these false alarms (Caseload). For each CDA, we plotted the Detection Delay as a function of FAR or Caseload, over a broad range of alarm thresholds. RESULTS: CDAs that combined text analyses seeking ARI symptoms in clinical notes with provider-assigned diagnostic codes in order to maximize the precision rather than the sensitivity of case-detection lowered Detection Delay at any given FAR or Caseload. CONCLUSION: An empiric approach can guide the integration of EMR data into case-detection methods that improve both the timeliness and efficiency of epidemic detection.


title: Coronavirus Disease 2019 (COVID‐19): Potential implications for weak health systems and conflict zones in the Middle East and North Africa region

text: This short communication recognizes the underbelly of weak and conflict‐prone health systems in the Middle East and North Africa region in the wake of COVID‐19 pandemic. The communication highlights how the lack of basic resources, absence of a well‐functioning health system and the dearth of well‐coordinated communication channels, can bode ill for the successful fight against COVID‐19. The article elucidates COVID‐19 potential health, social, and economic implications for such countries. The communication cautions that if COVID‐19 is left to incubate and makes a home in weak systems, it will have a much better chance of mutating and coming back to infect many people globally. The communication calls on the international institutions in collaboration with developed nations to be prepared to probe up health systems in weak and conflict‐prone health systems with much‐needed resources in order to nip COVID‐19 in the bud.


Shoulder & Elbow Education During COVID19 – Perspectives from the Resident, Fellow, and Attending Level