title: MERS-CoV spillover at the camel-human interface

text: Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic virus from camels causing significant mortality and morbidity in humans in the Arabian Peninsula. The epidemiology of the virus remains poorly understood, and while case-based and seroepidemiological studies have been employed extensively throughout the epidemic, viral sequence data have not been utilised to their full potential. Here, we use existing MERS-CoV sequence data to explore its phylodynamics in two of its known major hosts, humans and camels. We employ structured coalescent models to show that long-term MERS-CoV evolution occurs exclusively in camels, whereas humans act as a transient, and ultimately terminal host. By analysing the distribution of human outbreak cluster sizes and zoonotic introduction times, we show that human outbreaks in the Arabian peninsula are driven by seasonally varying zoonotic transfer of viruses from camels. Without heretofore unseen evolution of host tropism, MERS-CoV is unlikely to become endemic in humans.


title: MBL-associated serine proteases (MASPs) and infectious diseases

text: Abstract The lectin pathway of the complement system has a pivotal role in the defense against infectious organisms. After binding of mannan-binding lectin (MBL), ficolins or collectin 11 to carbohydrates or acetylated residues on pathogen surfaces, dimers of MBL-associated serine proteases 1 and 2 (MASP-1 and MASP-2) activate a proteolytic cascade, which culminates in the formation of the membrane attack complex and pathogen lysis. Alternative splicing of the pre-mRNA encoding MASP-1 results in two other products, MASP-3 and MAp44, which regulate activation of the cascade. A similar mechanism allows the gene encoding MASP-2 to produce the truncated MAp19 protein. Polymorphisms in MASP1 and MASP2 genes are associated with protein serum levels and functional activity. Since the first report of a MASP deficiency in 2003, deficiencies in lectin pathway proteins have been associated with recurrent infections and several polymorphisms were associated with the susceptibility or protection to infectious diseases. In this review, we summarize the findings on the role of MASP polymorphisms and serum levels in bacterial, viral and protozoan infectious diseases.


title: B cell characterization and reactivity analysis in multiple sclerosis

text: Abstract B cells are one of the key players in the pathogenesis of multiple sclerosis (MS). The peripheral B cell distributions are similar in healthy persons and MS patients. In healthy controls, B cells are rarely present in the cerebrospinal fluid (CSF) while in MS patients, a clonally expanded B cell population is detected. This consists of memory B cells, centroblasts and antibody-secreting plasma blasts and plasma cells that are responsible for intrathecal immunoglobulin G production and oligoclonal band formation in more than 90% of MS patients. Unfortunately, the targets of the autoreactive B cells and antibodies remain largely unknown. Various candidate antigens have been identified but often their involvement in the disease process is still unclear. Most studies characterizing these target antigens examined autoantibodies by analyzing sera or CSF of MS patients. An alternative approach is focusing on the clonally expanded B cells. In this way B cells directed against myelin, astroglia and axons have been denoted in MS patients. B cell immortalization, that is based on the antibody-producing potential of Epstein–Barr virus (EBV) transformed B cells, can be used to expand B cells from MS patients for the production of antibodies, that ultimately can be analysed for target identification.


title: Treatment of sleep apnea by ENT specialists during the COVID-19 pandemic

text: Abstract The treatment of sleep disorders has been strongly impacted by the COVID-19 pandemic. When the lockdown is over, resumption of usual patient care will require precautions to limit the risk of contamination for patients and caregivers. In this document, the French Association of Otorhinolaryngology and Sleep disorders (AFSORL) and the French Society of Otorhinolaryngology (SFORL) put forward a summary of the measures for continuing the treatment of sleep apnoea syndrome in these new practice conditions. Emphasis is placed on teleconsultation, methods of nocturnal sleep studies, the conditions for treatment by continuous positive airway pressure (CPAP) ventilation, and the postponement of more invasive treatments.


Nervous system damage after COVID-19 infection: presence or absence?


title: Use of DAMPs and SAMPs as Therapeutic Targets or Therapeutics: A Note of Caution

text: This opinion article discusses the increasing attention paid to the role of activating damage-associated molecular patterns (DAMPs) in initiation of inflammatory diseases and suppressing/inhibiting DAMPs (SAMPs) in resolution of inflammatory diseases and, consequently, to the future roles of these novel biomarkers as therapeutic targets and therapeutics. Since controlled production of DAMPs and SAMPs is needed to achieve full homeostatic restoration and repair from tissue injury, only their pathological, not their homeostatic, concentrations should be therapeutically tackled. Therefore, distinct caveats are proposed regarding choosing DAMPs and SAMPs for therapeutic purposes. For example, we discuss the need to a priori identify and define a context-dependent “homeostatic DAMP:SAMP ratio” in each case and a “homeostatic window” of DAMP and SAMP concentrations to guarantee a safe treatment modality to patients. Finally, a few clinical examples of how DAMPs and SAMPs might be used as therapeutic targets or therapeutics in the future are discussed, including inhibition of DAMPs in hyperinflammatory processes (e.g., systemic inflammatory response syndrome, as currently observed in Covid-19), administration of SAMPs in chronic inflammatory diseases, inhibition of SAMPs in hyperresolving processes (e.g., compensatory anti-inflammatory response syndrome), and administration/induction of DAMPs in vaccination procedures and anti-cancer therapy.


title: Molecular mimicry of ACTH in SARS – implications for corticosteroid treatment and prophylaxis

text: Abstract For a virus to survive and replicate in an organism, it must employ strategies to evade and misdirect the host's immune response. There is compelling evidence that the primary immunoevasive strategy utilized by the SARS virus, like influenza, is to inhibit its host's corticosteroid stress response. This is accomplished by viral expression of amino acid sequences that are molecular mimics of the host's adrenocorticotropin hormone (ACTH). When the host produces antibodies against these viral antigens, the antibodies also bind to the host's own ACTH, which limits the host's stress response by interfering with ACTH's ability to stimulate the secretion of corticosteroids. This inadequate corticosteroid response provokes symptoms as a result of a relative adrenocortical insufficiency. Treatment with corticosteroids can relieve the patient's symptoms of adrenocortical insufficiency and give them the corticosteroid levels needed to fight their infection. Similarly, by taking moderate daily doses of corticosteroids as a prophylactic, it may be possible to avoid clinical infection with SARS. If SARS's ACTH mimic strategy never has an opportunity to get started, SARS's ability to evade its host's immune system while its viral load is low will be significantly impaired. In this article, amino acid sequences from the SARS and influenza viruses representing likely homology to human ACTH are identified. Evidence demonstrating that ACTH autoantibodies are produced during influenza infection is also presented. Early treatment with corticosteroids should lower the dose necessary to counteract SARS's ACTH autoantibody mechanism. If corticosteroid treatment is delayed until inflammatory cytokine levels are causing serious injury, only high doses of corticosteroids are likely to be effective.


title: Corticosteroid therapy for patients with CoVID-19 pneumonia: a before-after study

text: BACKGROUND: Anti-inflammatory drugs such as corticosteroids may beneficially modulate the host inflammatory response to CoVID-19 pneumonia. AIMS: To evaluate the impact of addition of corticosteroids to the hospital protocol for treatment of suspected or confirmed CoVID-19 pneumonia on rates of death or intensive care unit (ICU) admission. METHODS: A before-after study was performed to evaluate the effect of addition of corticosteroids to our institution's COVID-19 treatment protocol on hospital mortality. RESULTS: Between March 3(rd) and April 14(th) 2020, 257 patients with CoVID-19 diagnosis were included. As corticosteroids were wide used since 27 March 2020, two periods were considered for the purposes of our study: the before period from March 3(rd) to 20(th) (n= 85) and the “after period” (n=172) from March 26(th) to April 14(th) 2020. The “after” period was associated with a lower risk of death (HR 0.47; 95% CI, 0.23 - 0.97; p=0.04), and a lower risk of intensive care admission or death before ICU admission (HR 0.37 95% CI 0.21 - 0.64; p=0.0005) by multivariate analysis adjusted for age, National Early Warning score and institutionalization status. CONCLUSIONS: In the “after period”, the addition of corticosteroids to our institution's CoVID-19 treatment protocol was associated with a significant reduction in hospital mortality.


title: Development of monoclonal antibodies and serological assays including indirect ELISA and fluorescent microsphere immunoassays for diagnosis of porcine deltacoronavirus

text: BACKGROUND: A novel porcine deltacoronavirus (PDCoV), also known as porcine coronavirus HKU15, was reported in China in 2012 and identified in the U.S. in early 2014. Since then, PDCoV has been identified in a number of U.S. states and linked with clinical disease including acute diarrhea and vomiting in the absence of other identifiable pathogens. Since PDCoV was just recently linked with clinical disease, few specific antibody-based reagents were available to assist in diagnosis of PDCoV and limited serological capabilities were available to detect an antibody response to this virus. Therefore, the overall objective of this project was to develop and validate selected diagnostic reagents and assays for PDCoV antigen and antibody detection. RESULTS: The nucleoprotein of PDCoV was expressed as a recombinant protein and purified for use as an antigen to immunize mice for polyclonal, hyperimmune sera and monoclonal antibody (mAb) production. The resulting mAbs were evaluated for use in fluorescent antibody staining methods to detect PDCoV infected cells following virus isolation attempts and for immunohistochemistry staining of intestinal tissues of infected pigs. The same antigen was used to develop serological tests to detect the antibody response to PDCoV in pigs following infection. Serum samples from swine herds with recent documentation of PDCoV infection and samples from expected naïve herds were used for initial assay optimization. The tests were optimized in a checkerboard fashion to reduce signal to noise ratios using samples of known status. Statistical analysis was performed to establish assay cutoff values and assess diagnostic sensitivities and specificities. At least 629 known negative serum samples and 311 known positive samples were evaluated for each assay. The enzyme linked immunosorbent assay (ELISA) showed diagnostic sensitivity (DSe) of 96.1 % and diagnostic specificity (DSp) of 96.2 %. The fluorescent microsphere immunoassay (FMIA) showed a DSe of 95.8 % and DSp of 98.1 %. Both ELISA and FMIA detected seroconversion of challenged pigs between 8–14 days post-infection (DPI). An indirect fluorescent antibody (IFA) test was also developed using cell culture adapted PDCoV for comparative purposes. CONCLUSION: These new, specific reagents and serological assays will allow for improved diagnosis of PDCoV. Since many aspects of PDCoV infection and transmission are still not fully understood, the reagents and assays developed in this project should provide valuable tools to help understand this disease and to aid in the control and surveillance of porcine deltacoronavirus outbreaks.


title: Analysis of long non-coding RNAs in neonatal piglets at different stages of porcine deltacoronavirus infection

text: BACKGROUND: PDCoV (Porcine Deltacoronavirus) is a novel porcine coronavirus that causes intestinal necrosis of piglets, thinning of the intestinal wall and severe villus atrophy in the small intestine. PDCoV is a highly contagious infectious disease characterized by diarrhea, dehydration and vomiting. It has been reported that lncRNA has a significant effect on viral replication and increased or decreased virulence. At present, there is almost no research on lncRNA related to PDCoV infection. With the development of the research, a large number of lncRNAs related to PDCoV infection have been discovered. Identifying the role of these lncRNAs in the infection process facilitates the screening of diagnostically significant biomarkers. RESULTS: Using high throughput sequencing to screen differentially expressed long non-coding RNA (lncRNA) during PDCoV infection, we identified 99, 41 and 33 differentially expressed lncRNAs in the early, middle and late stages of infection, respectively. These lncRNAs were involved in glycolysis / gluconeogenesis, histidine metabolism and pentose and Chloroalkane and chloroalkene degradation pathway. We obtained expression data of miRNAs, lncRNAs and mRNAs during PDCoV infection and constructed and investigated an interaction network. The qRT-PCR validation results of 6 differentially expressed lncRNAs were consistent with RNA-Seq results. CONCLUSIONS: This study is the first to examine differentially expressed lncRNAs after PDCoV infection of piglets. These results can provide new insights into PDCoV infection and antiviral strategies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-019-1862-4) contains supplementary material, which is available to authorized users.


title: Assembling the prenylneoflavone system through a Pechmann condensation/Mitsunobu reaction/Claisen rearrangement/olefin cross-metathesis sequence

text: ABSTRACT: The multistep synthesis of a prenylneoflavone through a sequence of the Mitsunobu reaction/Claisen rearrangement/olefin cross-metathesis reaction has been accomplished in 5% yield over six steps starting from commercially available 3-methoxyacetophenone. The sequence is shown to be compatible with a Pechmann condensation which proved to be a robust and cost-effective method for the assembling of the α-pyrone core. The results open doors to a general approach to the prenylneoflavone system starting from phenol and acetophenone derivatives. GRAPHIC ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00706-020-02584-8) contains supplementary material, which is available to authorized users.


Telehealth: An Important Player During the COVID-19 Pandemic


title: Small Business Administration emergency loans available for nonprofits

text: In its effort to minimize economic disruption wrought by COVID‐19, the U.S. Small Business Administration is working directly with state governors to provide targeted, low‐interest disaster recovery loans to small businesses and private nonprofit organizations that have been severely impacted by the pandemic.


title: Challenges in COVID‐19 medical response: a nephrology perspective

text: The new coronavirus disease, named by World Health Organization (WHO) as COVID‐19 brought great challenges to patients with end‐stage renal disease (ESRD). In general, ESRD patients have higher number of comorbidities and are at age‐risk for severe pulmonary presentation of this disease. Another important issue is that hemodialysis (HD) clinics are usually not located in small towns, and these frail patients often travel to their dialysis center in groups and also cannot keep the 6‐feet safe distance during their HD session.(1)


title: Feasibility of Long-Term Proteasome Inhibition in Multiple Myeloma by In-Class Transition from Bortezomib to Ixazomib

text: BACKGROUND: The ongoing US MM-6 study is investigating in-class transition (iCT) from parenteral bortezomib-based induction to all-oral ixazomib-lenalidomide-dexamethasone (ixazomib-Rd) with the aim of increasing proteasome inhibitor (PI)-based treatment adherence/duration while maintaining quality of life (QoL) and improving outcomes. PATIENTS AND METHODS: US community sites are enrolling non-transplant-eligible newly diagnosed multiple myeloma (MM) patients with no evidence of progressive disease after 3 cycles of bortezomib-based therapy to receive ixazomib-Rd (up to 39 cycles or until progression/toxicity). Patients use mobile/wearable digital devices to collect actigraphy (activity/sleep) data and electronically complete QoL/treatment satisfaction/medication adherence questionnaires. Primary endpoint: progression-free survival (PFS); key secondary endpoints include response rates and therapy duration. RESULTS: At data cutoff, 84 patients had been treated (median age 73 years; 44% ≥ 75 years; 49% male; 15% black/African American; 10% Hispanic/Latino); 62% of patients remain on therapy. Mean duration of total PI therapy was 10.1 months and of ixazomib-Rd was 7.3 months. With 8 months median follow-up, 12-month PFS rate was 86% (95% confidence interval, 73–93) from both the start of bortezomib-based treatment and the start of ixazomib-Rd. Overall response rate was 62% (complete response [CR], 4%; very good partial response [VGPR], 25%; partial response [PR], 33%) after bortezomib-based induction and 70% (CR, 26%; VGPR, 29%; PR, 15%) after iCT. The ixazomib-Rd safety profile was consistent with previous clinical trial data. QoL/treatment satisfaction were maintained. CONCLUSION: US MM-6 patients are representative of the real-world US MM population; iCT may permit prolonged PI-based therapy with promising efficacy, without impacting patients’ QoL/treatment satisfaction.


title: Biodiversity and clinico-demographic characteristics of human rhinoviruses from hospitalized children with acute lower respiratory tract infections in Malaysia

text: Abstract Background There is accumulating evidence that human rhinovirus (HRV) causes acute lower respiratory tract infections (ALRTI). Recently, HRV-C was identified as a new species of HRV, but its spectrum of clinical disease is not well understood. Objectives We investigated the molecular epidemiology, demographic and clinical characteristics of HRVs among hospitalized children with ALRIs. Study design One hundred and sixty-five nasopharangeal aspirates taken from children <5 years hospitalized with ALRTIs in Serdang Hospital, Malaysia, were subject to reverse transcriptase-PCR for HRV. Phylogenetic analysis on VP4/VP2 and 5′-NCR regions was used to further characterize HRV. Other respiratory viruses were also investigated using semi-nested multiplex RT-PCR assay. Clinical parameters were analyzed between HRV, RSV and IFV-A mono-infections and between HRV species. Results HRV was detected in 54 (33%) patients for both single (36 samples) and multiple (18 samples) infections, 61.1% (22/36) represents HRV-A strains while the remaining 14 HRV-C. Strain P51was the first reported representative of HRV98. The majority of the single HRV cases were in the second half of infancy; HRV-C occurred among older children compared with HRV-A. HRV children were admitted significantly earlier and less febrile than RSV and IFV-A infection. HRV-C infected children were more likely to have rhonchi and vomiting as compared to HRV-A. Pneumonia was the most common discharge diagnosis followed by bronchiolitis and post-viral wheeze in HRV patients. Conclusion Our study showed high prevalence of HRVs and detection of HRV-C among hospitalized children with ALRTIs in Malaysia. Analysis of clinical parameters suggested specific features associated with HRVs infections and specific HRV groups.


title: Influenza

text: Zusammenfassung Virus Influenzavirus ist ein Mitglied der Orthomyxoviren, welche in der Evolution die Fähigkeit erworben haben, schnelle Wirtswechsel vorzunehmen. Typische Wirte sind Mensch, Haustiere wie Pferd, Schwein, Katze und Geflügel, unter den Vögeln besonders Gans und Ente, und Säugetiere des Wassers. Das Genom von Influenzavirus A und B ist segmentiert und wenn zwei Viren eine Zelle infizieren können ganze Genomsegmente ausgetauscht werden (reassortment), was zu einer hohen Variabilität führt. Diese betrifft sowohl die Oberflächenproteine der Hülle des Influenzavirus wie H (Hämagglutinin) und N (Neuraminidase) als auch die im Partikel liegende Polymerase. Ein Influenzavirus, welches auf der Oberfläche Epitope trägt, die dem tierischen oder menschlichen Immunsystem nicht bekannt sind, hat das Potential eine neue Epidemie auszulösen. Übertragung typisch für Influenzavirus ist die aerogene Übertragung, besonders über Tröpfchen beim Husten und Niesen und Schmierinfektion, meist durch direkten Kontakt. Die Inkubationszeit beträgt normalerweise 24 bis 48 Stunden. Frühe klinische Symptome sind Fieber, Infektionszeichen (Halsschmerzen, Nasensekretion, Niesen) im oberen Respirationstrakt, Augenrötung gepaart mit Myalgie, Schüttelfrost und schwerem Krankheitsgefühl. Prävention wesentlich sind Abstand vom Infizierten, Vermeiden von Aerosolen und - innerhalb der Behandlungseinheiten - Händedesinfektion und Desinfektion von Gebrauchsgegenständen vom Stethoskop bis zur Türklinke. Ein wesentlicher Beitrag zur Prävention wird durch die jährliche Impfung gegen saisonale Stämme Influenzavirus A-H1N1 und A-H3N2 und Influenzavirus B geleistet. Therapie unspezifisch entzündungshemmende Substanzen und Vermeiden bzw. Behandlung der bakteriellen Superinfektion mit Antibiotika. Spezifisch mit Neuraminidase-Inhibitoren wie Oseltamivir (Tamiflu®) und Zanamivir (Relenza®) innerhalb von 48h nach Beginn der Symptomatik. Abstract Virus Influenza virus is the only human-pathogenic member of orthomyxovirus, which during evolution acquired the capability to infect the cells of various mammals and birds. Besides man common hosts are domestic animals as horse, swine, cat and poultry, among the birds especially goose and duck, and nearly all mammals living in water. The influenza virus A and B genome is segmented, when two different viruses infect one cell whole genome-segments can be exchanged (reassortment). This capability leads to a high variability of the envelope surface proteins H (haemagglutinin) and N (neuraminidase), as well as the polymerase inside the virus particle. An influenza virus equipped with epitopes on its surface that are unknown by the human and animal immune system has the potential to initiate a new epidemic. Transmission for influenza virus main route is aerogene, especially by small droplets while coughing and sneezing and by smear infection, mostly by direct contact. Incubation time is usually 24 to 48hours. Early clinical symptoms are fever, all signs of inflammation in the upper respiratory tract with sore throat, nasal secretion, conjunctivitis that are associated with myalgia, chills and malaise. Prevention important is the distance from the infected, to avoid aerosol formation, and within care units disinfection of hands and utensils as from stethoscope to door handle. Essential for prevention of disease is the annual vaccination against the seasonal strains influenza virus A-H1N1, A-H3N2 and influenza virus B. Treatment unspecific by anti-inflammatory drugs and reduction of symptoms by therapy of bacterial superinfection with antibiotics. Specific: by application of neuraminidase inhibitors as oseltamivir (Tamiflu®) and zanamavir (Relenza®) within 48hours after onset of the symptoms.


title: The need for fully bio-based facemasks to counter coronavirus outbreaks: A perspective

text: Abstract The onset of coronavirus pandemic has sparked a shortage of facemasks in almost all nations. Without this personal protective equipment, healthcare providers, essential workers, and the general public are exposed to the risk of infection. In light of the aforementioned, it is critical to balance the supply and demand for masks. COVID-19 will also ensure that masks are always considered as an essential commodity in future pandemic preparedness. Moreover, billions of facemasks are produced from petrochemicals derived raw materials, which are non-degradable upon disposal after their single use, thus causing environmental pollution and damage. The sustainable way forward is to utilise raw materials that are side-stream products of local industries to develop facemasks having equal or better efficiency than the conventional ones. In this regard, wheat gluten biopolymer, which is a by-product or co-product of cereal industries, can be electrospun into nanofibre membranes and subsequently carbonised at over 700 °C to form a network structure, which can simultaneously act as the filter media and reinforcement for gluten-based masks. In parallel, the same gluten material can be processed into cohesive thin films using plasticiser and hot press. Additionally, lanosol, a naturally-occurring substance, imparts fire (V-0 rating in vertical burn test), and microbe resistance in gluten plastics. Thus, thin films of flexible gluten with very low amounts of lanosol (<10 wt%) can be bonded together with the carbonised mat and shaped by thermoforming to create the facemasks. The carbon mat acting as the filter can be attached to the masks through adapters that can also be made from injection moulded gluten. The creation of these masks could simultaneously be effective in reducing the transmittance of infectious diseases and pave the way for environmentally benign sustainable products.


title: Protection against infectious bronchitis virus by spike ectodomain subunit vaccine

text: Abstract The avian coronavirus infectious bronchitis virus (IBV) S1 subunit of the spike (S) glycoprotein mediates viral attachment to host cells and the S2 subunit is responsible for membrane fusion. Using IBV Arkansas-type (Ark) S protein histochemistry, we show that extension of S1 with the S2 ectodomain improves binding to chicken tissues. Although the S1 subunit is the major inducer of neutralizing antibodies, vaccination with S1 protein has been shown to confer inadequate protection against challenge. The demonstrated contribution of S2 ectodomain to binding to chicken tissues suggests that vaccination with the ectodomain might improve protection compared to vaccination with S1 alone. Therefore, we immunized chickens with recombinant trimeric soluble IBV Ark-type S1 or S-ectodomain protein produced from codon-optimized constructs in mammalian cells. Chickens were primed at 12days of age with water-in-oil emulsified S1 or S-ectodomain proteins, and then boosted 21days later. Challenge was performed with virulent Ark IBV 21days after boost. Chickens immunized with recombinant S-ectodomain protein showed statistically significantly (P <0.05) reduced viral loads 5days post-challenge in both tears and tracheas compared to chickens immunized with recombinant S1 protein. Consistent with viral loads, significantly reduced (P <0.05) tracheal mucosal thickness and tracheal lesion scores revealed that recombinant S-ectodomain protein provided improved protection of tracheal integrity compared to S1 protein. These results indicate that the S2 domain has an important role in inducing protective immunity. Thus, including the S2 domain with S1 might be promising for better viral vectored and/or subunit vaccine strategies.


Post-containment management of nursing homes: a new public health concern


title: Covid-19 y la educación médica: retos y oportunidades en venezuela

text: ABSTRACT On 11 March 2020 the World Health Organisation declared the Pandemic situation due to a new coronavirus, Severe Acute Respiratory Syndrome -Coronavirus 2 (SARS-COV-2), which caused the infectious disease due to coronavirus called COVID-19. University education in Venezuela, and particularly medical education, due to its close relationship with the Pandemic, has been severely affected. A review is presented in this article on the available evidence and opinions as regards medical education in the COVID-19 era. A series of challenges are set out which the University may face from the reflection and the establishing of solutions in a context of uncertainty and accelerated changes for the future of Medical Education.The outlook for Medical Education in the country and at world level is uncertain, and although unfavourable at the moment, the evolutionary process not only offers adverse environments, but also the possibility of generating adaptive processes, and the search and implementation of new ways of teaching. This latter, may involve a positive and necessary change for the teaching of new health professionals.


title: Mental Health During the Covid-19 Outbreak in China: a Meta-Analysis

text: Background: Covid-19 has started to spread within China since the end of December 2019. Despite government’s immediate actions and strict control, more and more people were infected every day. As such a contagious virus can spread easily and rapidly between people, the whole country was put into lockdown and people were forced into isolation. In order to understand the impact of Covid-19 on mental health well-being, Chinese researchers have conducted several studies. However, no consistent results were obtained. Therefore, a meta-analysis was conducted. Methods: We searched Embase, PubMed, and Web of Science databases to find literature from December 2019 to April 2020 related to Covid-19 and mental health, among which results such as comments, letters, reviews and case reports were excluded. The incidence of anxiety and depression in the population was synthesized and discussed. Results: A total of 27,475 subjects were included in 12 studies. Random effect model is used to account for the data. The results showed that the incidence of anxiety was 25% (95% CI: 0.19–0.32), and the incidence of depression was 28% (95% CI: 0.17–0.38). Significant heterogeneity was detected across studies regarding these incidence estimates. Subgroup analysis included the study population and assessment tools, and sensitivity analysis was done to explore the sources of heterogeneity. Conclusions: Owing to the significant heterogeneity detected in studies regarding this pooled prevalence of anxiety and depression, we must interpret the results with caution. As the epidemic is ongoing, it is vital to set up a comprehensive crisis prevention system, which integrating epidemiological monitoring, screening and psychological crisis prevention and interventions.


title: Comparative analysis of four Massachusetts type infectious bronchitis coronavirus genomes reveals a novel Massachusetts type strain and evidence of natural recombination in the genome

text: Abstract Four Massachusetts-type (Mass-type) strains of infectious bronchitis coronavirus (IBV) were compared genetically with the pathogenic M41 and H120 vaccine strains using the complete genomic sequences. The results revealed that strains ck/CH/LNM/091017 and ck/CH/LDL/101212 were closely related to the H120 vaccine, which suggests that they might represent re-isolations of vaccine strains or variants of vaccine strains that have resulted from the accumulated point mutations after several passages in chickens. In contrast, strains ck/CH/LHLJ/07VII and ck/CH/LHLJ/100902 had a close genetic relationship with the pathogenic M41 strain. In addition, molecular markers have been identified that distinguish between field and vaccine (or vaccine-like) Mass-type viruses, which may be able to differentiate between field and vaccine strains for diagnostic purposes. Phylogenetic analysis, and pairwise comparison of full-length genomes and the nine genes, identified the occurrence of recombination events in the genome of strain CK/VH/LHLJ/07VII, which suggests that this virus originated from recombination events between M41- and H120-like strains at the switch site located at the 3′ end of the nucleocapsid (N) genes. To our knowledge, this is the first time that evidence for the evolution and natural recombination under field conditions between Mass-type pathogenic and vaccinal IBV strains has been documented. These findings provide insights into the emergence and evolution of the Mass-type IB coronaviruses and may help to explain the emergence of Mass-type IBV in chicken flocks all over the world.


title: Conflicts of Interest during Contact Investigations: A Game-Theoretic Analysis

text: The goal of contact tracing is to reduce the likelihood of transmission, particularly to individuals who are at greatest risk for developing complications of infection, as well as identifying individuals who are in need of medical treatment of other interventions. In this paper, we develop a simple mathematical model of contact investigations among a small group of individuals and apply game theory to explore conflicts of interest that may arise in the context of perceived costs of disclosure. Using analytic Kolmogorov equations, we determine whether or not it is possible for individual incentives to drive noncooperation, even though cooperation would yield a better group outcome. We found that if all individuals have a cost of disclosure, then the optimal individual decision is to simply not disclose each other. With further analysis of (1) completely offsetting the costs of disclosure and (2) partially offsetting the costs of disclosure, we found that all individuals disclose all contacts, resulting in a smaller basic reproductive number and an alignment of individual and group optimality. More data are needed to understand decision making during outbreak investigations and what the real and perceived costs are.


title: Inhibitory and Coactivating Receptors Recognising the Same Ligand: Immune Homeostasis Exploited by Pathogens and Tumours

text: Coactivating and inhibitory receptors that share at least one ligand interact with a wide variety of ligands, indicating their importance in a range of situations. Here, we discuss principles of mainly human paired receptor function and ligand recognition, and possible therapeutic implications of targeting these receptors in cancer, autoimmune diseases, and allergy. We summarise and emphasise the idea that these receptors, which have evolved in part in response to pathogen pressure, fine-tune the immune response, preserve homeostasis, and that pathogens and tumours use the dominance of the inhibitory receptors over the coactivating receptors to avoid immune elimination. Finally, we discuss the options of using paired receptors and their ligand for immune cell education and therapy.


title: Human Pulmonary 3D Models For Translational Research

text: Lung diseases belong to the major causes of death worldwide. Recent innovative methodological developments now allow more and more for the use of primary human tissue and cells to model such diseases. In this regard, the review covers bronchial air‐liquid interface cultures, precision cut lung slices as well as ex vivo cultures of explanted peripheral lung tissue and de‐/re‐cellularization models. Diseases such as asthma or infections are discussed and an outlook on further areas for development is given. Overall, the progress in ex vivo modeling by using primary human material could make translational research activities more efficient by simultaneously fostering the mechanistic understanding of human lung diseases while reducing animal usage in biomedical research.


title: Vaccine platform recombinant measles virus

text: The classic development of vaccines is lengthy, tedious, and may not necessarily be successful as demonstrated by the case of HIV. This is especially a problem for emerging pathogens that are newly introduced into the human population and carry the inherent risk of pandemic spread in a naïve population. For such situations, a considerable number of different platform technologies are under development. These are also under development for pathogens, where directly derived vaccines are regarded as too complicated or even dangerous due to the induction of inefficient or unwanted immune responses causing considerable side-effects as for dengue virus. Among platform technologies are plasmid-based DNA vaccines, RNA replicons, single-round infectious vector particles, or replicating vaccine-based vectors encoding (a) critical antigen(s) of the target pathogens. Among the latter, recombinant measles viruses derived from vaccine strains have been tested. Measles vaccines are among the most effective and safest life-attenuated vaccines known. Therefore, the development of Schwarz-, Moraten-, or AIK-C-strain derived recombinant vaccines against a wide range of mostly viral, but also bacterial pathogens was quite straightforward. These vaccines generally induce powerful humoral and cellular immune responses in appropriate animal models, i.e., transgenic mice or non-human primates. Also in the recent first clinical phase I trial, the results have been quite encouraging. The trial indicated the expected safety and efficacy also in human patients, interestingly independent from the level of prevalent anti-measles immunity before the trial. Thereby, recombinant measles vaccines expressing additional antigens are a promising platform for future vaccines.


title: Endoplasmic Reticulum: The Favorite Intracellular Niche for Viral Replication and Assembly

text: The endoplasmic reticulum (ER) is the largest intracellular organelle. It forms a complex network of continuous sheets and tubules, extending from the nuclear envelope (NE) to the plasma membrane. This network is frequently perturbed by positive-strand RNA viruses utilizing the ER to create membranous replication factories (RFs), where amplification of their genomes occurs. In addition, many enveloped viruses assemble progeny virions in association with ER membranes, and viruses replicating in the nucleus need to overcome the NE barrier, requiring transient changes of the NE morphology. This review first summarizes some key aspects of ER morphology and then focuses on the exploitation of the ER by viruses for the sake of promoting the different steps of their replication cycles.


title: Oral immunization of cattle with hemagglutinin protein of rinderpest virus expressed in transgenic peanut induces specific immune responses

text: Rinderpest is an acute, highly contagious often fatal disease of large and small ruminants, both domestic and wild. Global eradication of rinderpest needs a robust, safe and cost-effective vaccine. The causative agent, rinderpest virus (RPV) is an important member of the genus Morbillivirus in the Paramyxoviridae family. We have generated transgenic peanut (Arachis hypogea L.) plants expressing hemagglutinin protein of RPV and report here, the induction of immune responses in cattle following oral feeding with transgenic leaves expressing hemagglutinin protein without oral adjuvant. Hemagglutinin-specific antibody was detected in the serum as confirmed by immunohistochemical staining of virus-infected cells, and in vitro neutralization of virus infectivity. Oral delivery also resulted in cell-mediated immune responses.


title: Tetraspanins: Architects of Viral Entry and Exit Platforms

text: Host factors render cells susceptible to viral infection. One family of susceptibility factors, the tetraspanin proteins, facilitate enveloped virus entry by promoting virus-cell membrane fusion. They also facilitate viral egress from infected cells. In this Gem, we discuss recent insights into how tetraspanins assemble viral entry and exit platforms on cell membranes, and we speculate that tetraspanins contribute to nonviral membrane fusions by similar mechanisms.


title: The Impact of the COVID-19 Pandemic on Fellowship Training: A National Survey of Pediatric Otolaryngology Fellowship Directors

text: OBJECTIVES: To gain a better understanding of the effects the COVID-19 pandemic has had on current and future pediatric otolaryngology fellowship training, as well as how the application process was impacted this past year. METHODS: An anonymous web-based survey consisting of 24 questions was sent to all fellowship directors. The survey questions were designed to gain a better understanding of the effects of the current COVID-19 pandemic on the surgical and clinical experience of current, to characterize the types of supplemental educational experiences that fellowship directors had incorporated into the curriculum to compensate for the decreased surgical and clinical workload, and highlight differences based on geographic location. RESULTS: Overall, 22 of 36 fellowship directors responded to our survey, for a total response rate of 61%. The Midwest had the highest response rate at 72.7%, followed by the Northeast (71.4%), the West (50%), and the South (50%). The vast majority of fellowship directors (77.2%) reported the COVID-19 pandemic had a “significant impact” on overall pediatric otolaryngology fellowship training. 86.3% of fellowship directors reported that their programs were still performing some surgical operations, but with decreased overall volume. Interestingly, 13.6% of fellowship directors reported that their fellows had been pulled to medicine or ICU services to assist with the COVID-19 pandemic. Of these programs that had a fellow pulled to the ICU or medicine service, 2 out of 3 were located in the Northeast, with the remaining fellow being from a program in the South. CONCLUSION: Overall, pediatric otolaryngology fellowship directors reported the COVID-19 pandemic has had a significant impact on the overall fellowship experience within the field of pediatric otolaryngology, with the majority feeling that both their fellows surgical and clinical experience have been significantly impacted.


title: COVID-19 and the Future of Microfinance: Evidence and Insights from Pakistan

text: The COVID-19 pandemic threatens lives and livelihoods, and, with that, has created immediate challenges for institutions that serve affected communities. We focus on implications for local microfinance institutions in Pakistan, a country with a mature microfinance sector, serving a large number of households. The institutions serve populations poorly-served by traditional commercial banks, helping customers invest in microenterprises, save, and maintain liquidity. We report results from ‘rapid response’ phone surveys of about 1,000 microenterprise owners, a survey of about 200 microfinance loan officers, and interviews with regulators and senior representatives of microfinance institutions. We ran these surveys starting about a week after the country went into lockdown to prevent the spread of the novel coronavirus. We find that, on average, week-on-week sales and household income both fell by about 90%. Households’ primary immediate concern in early April became how to secure food. As a result, 70% of the sample of current microfinance borrowers reported that they could not repay their loans; loan officers anticipated a repayment rate of just 34% in April 2020. We build from the results to argue that COVID-19 represents a crisis for microfinance in low-income communities. It is also a chance to consider the future of microfinance, and we suggest insights for policy reform.


title: Complement and its role in protection and pathogenesis of flavivirus infections

text: The complement system is a family of serum and cell surface proteins that recognize pathogen-associated molecular patterns, altered-self ligands, and immune complexes. Activation of the complement cascade triggers several antiviral functions including pathogen opsonization and/or lysis, and priming of adaptive immune responses. In this review, we will examine the role of complement activation in protection and/or pathogenesis against infection by Flaviviruses, with an emphasis on experiments with West Nile and Dengue viruses.


title: Development of reverse transcription loop-mediated isothermal amplification for rapid detection of porcine epidemic diarrhea virus

text: In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for detection of porcine epidemic diarrhea virus (PEDV). Six primers were designed to amplify the nucleocapsid (N) gene of PEDV. The optimization, sensitivity, and specificity of the RT-LAMP were investigated. The results showed that the optimal reaction condition for RT-LAMP amplifying PEDV N gene was achieved at 63°C for 50 min. The RT-LAMP assay was more sensitive than gel-based RT-PCR and enzyme-linked immunosorbent assay. It was capable of detecting PEDV from clinical samples and differentiating PEDV from Porcine transmissible gastroenteritis virus, Porcine rotavirus, Porcine pseudorabies virus, Porcine reproductive and respiratory syndrome virus, and Avian infectious bronchitis virus.


title: Molecular modeling and analysis of hepatitis E virus (HEV) papain-like cysteine protease

text: Abstract The biochemical or biophysical characterization of a papain-like cysteine protease in HEV ORF1-encoded polyprotein still remains elusive. Very recently, we have demonstrated the indispensability of ORF1 protease-domain cysteines and histidines in HEV replication, ex vivo (Parvez, 2013). In this report, the polyprotein partial sequences of HEV strains and genetically-related RNA viruses were analyzed, in silico. Employing the consensus-prediction results of RUBV-p150 protease as structural-template, a 3D model of HEV-protease was deduced. Similar to RUBV-p150, a ‘papain-like β-barrel fold’ structurally confirmed the classification of HEV-protease. Further, we recognized a catalytic ‘Cys434-His443’ dyad homologue of RUBV-p150 (Cys1152-His1273) and FMDV-Lpro (Cys51-His148) in line with our previous mutational analysis that showed essentiality of ‘His443’ but not ‘His590’ in HEV viability. Moreover, a RUBV ‘Zn2+ binding motif’ (Cys1167-Cys1175-Cys1178-Cys1225-Cys1227) equivalent of HEV was identified as ‘Cys457-His458-Cys459 and Cys481-Cys483’ residues within the ‘β-barrel fold’. Notably, unlike RUBV, ‘His458’ also clustered therein, that was in conformity with the consensus cysteine protease ‘Zn2+-binding motif’. By homology, we also proposed an overlapping ‘Ca2+-binding site’ ‘D-X-[DNS]-[ILVFYW]-[DEN]-G-[GP]-XX-DE’ signature, and a ‘proline-rich motif’ interacting ‘tryptophan (W437-W472)’ module in the modeled structure. Our analysis of the predicted model therefore, warrants critical roles of the ‘catalytic dyad’ and ‘divalent metal-binding motifs’ in HEV protease structural-integrity, ORF1 self-processing, and RNA replication. This however, needs further experimental validations.


title: Solution-phase synthesis of 2-cyano and 2-amido aziridinyl peptides

text: Abstract Starting from a library of 2-l-α-amino acyl (E)-acrylonitriles, different short 2-cyano and 2-amido aziridinyl peptides, potential protease inhibitors, were obtained under parallel solution-phase conditions. The transformations include careful selection of conditions for aziridine deprotection and cyano group partial hydrolysis.


title: Viral aetiology of common colds of outpatient children at primary care level and the use of antibiotics

text: Although antibiotics are ineffective against viral respiratory infections, studies have shown high rates of prescriptions worldwide. We conducted a study in Brazil to determine the viral aetiologies of common colds in children and to describe the use of antibiotics for these patients. Children up to 12 years with common colds were enrolled from March 2008-February 2009 at a primary care level facility and followed by regular telephone calls and medical consultations. A nasopharyngeal wash was obtained at enrollment and studied by direct fluorescence assay and polymerase chain reaction for nine different types of virus. A sample of 134 patients was obtained, median age 2.9 years (0.1-11.2 y). Respiratory viruses were detected in 73.9% (99/134) with a coinfection rate of 30.3% (30/99). Rhinovirus was the most frequent virus (53/134; 39.6%), followed by influenza (33/134; 24.6%) and respiratory syncytial virus (8/134; 13.4%). Antibiotic prescription rate was 39.6% (53/134) and 69.8% (37/53) were considered inappropriate. Patients with influenza infection received antibiotics inappropriately in a greater proportion of cases when compared to respiratory syncytial virus and rhinovirus infections (p = 0.016). The rate of inappropriate use of antibiotics was very high and patients with influenza virus infection were prescribed antibiotics inappropriately in a greater proportion of cases.


title: The Diagnosis and Treatment of Elderly Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease and Chronic Bronchitis

text: The syndrome of chronic obstructive pulmonary disease (COPD) consists of chronic bronchitis (CB), bronchiectasis, emphysema, and reversible airway disease that combine uniquely in an individual patient. Older patients are at risk for COPD and its components—emphysema, CB, and bronchiectasis. Bacterial and viral infections play a role in acute exacerbations of COPD (AECOPD) and in acute exacerbations of CB (AECB) without features of COPD. Older patients are at risk for resistant bacterial organisms during their episodes of AECOPD and AECB. Organisms include the more‐common bacteria implicated in AECOPD/AECB such as Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. Less‐common nonenteric, gram‐negative organisms including Pseudomonas aeruginosa, gram‐positive organisms including Staphylococcus aureus, and strains of nontuberculosis Mycobacteria are more often seen in AECOPD/AECB episodes involving elderly patients with frequent episodes of CB or those with bronchiectasis. Risk‐stratified antibiotic treatment guidelines appear useful for purulent episodes of AECOPD and episodes of AECB. These guidelines have not been prospectively validated for the general population and especially not for the elderly population. Using a risk‐stratification approach for elderly patients, first‐line antibiotics (e.g., amoxicillin, ampicillin, pivampicillin, trimethoprim/sulfamethoxazole, and doxycycline), with a more‐limited spectrum of antibacterial coverage, are used in patients who are likely to have a low probability of resistant organisms during AECOPD/AECB. Second‐line antibiotics (e.g., amoxicillin/clavulanic acid, second‐ or third‐generation cephalosporins, and respiratory fluoroquinolones) with a broader spectrum of coverage are reserved for patients with significant risk factors for resistant organisms and those who have failed initial antibiotic treatment.


title: Molecular Basis for ADP-Ribose Binding to the Mac1 Domain of SARS-CoV-2 nsp3

text: [Image: see text] The virus that causes COVID-19, SARS-CoV-2, has a large RNA genome that encodes numerous proteins that might be targets for antiviral drugs. Some of these proteins, such as the RNA-dependent RNA polymerase, helicase, and main protease, are well conserved between SARS-CoV-2 and the original SARS virus, but several others are not. This study examines one of the proteins encoded by SARS-CoV-2 that is most different, a macrodomain of nonstructural protein 3 (nsp3). Although 26% of the amino acids in this SARS-CoV-2 macrodomain differ from those observed in other coronaviruses, biochemical and structural data reveal that the protein retains the ability to bind ADP-ribose, which is an important characteristic of beta coronaviruses and a potential therapeutic target.


title: Variation in False-Negative Rate of Reverse Transcriptase Polymerase Chain Reaction–Based SARS-CoV-2 Tests by Time Since Exposure

text: BACKGROUND: Tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) based on reverse transcriptase polymerase chain reaction (RT-PCR) are being used to “rule out” infection among high-risk persons, such as exposed inpatients and health care workers. It is critical to understand how the predictive value of the test varies with time from exposure and symptom onset to avoid being falsely reassured by negative test results. OBJECTIVE: To estimate the false-negative rate by day since infection. DESIGN: Literature review and pooled analysis. SETTING: 7 previously published studies providing data on RT-PCR performance by time since symptom onset or SARS-CoV-2 exposure using samples from the upper respiratory tract (n = 1330). PATIENTS: A mix of inpatients and outpatients with SARS-CoV-2 infection. MEASUREMENTS: A Bayesian hierarchical model was fitted to estimate the false-negative rate by day since exposure and symptom onset. RESULTS: Over the 4 days of infection before the typical time of symptom onset (day 5), the probability of a false-negative result in an infected person decreases from 100% (95% CI, 100% to 100%) on day 1 to 67% (CI, 27% to 94%) on day 4. On the day of symptom onset, the median false-negative rate was 38% (CI, 18% to 65%). This decreased to 20% (CI, 12% to 30%) on day 8 (3 days after symptom onset) then began to increase again, from 21% (CI, 13% to 31%) on day 9 to 66% (CI, 54% to 77%) on day 21. LIMITATION: Imprecise estimates due to heterogeneity in the design of studies on which results were based. CONCLUSION: Care must be taken in interpreting RT-PCR tests for SARS-CoV-2 infection—particularly early in the course of infection—when using these results as a basis for removing precautions intended to prevent onward transmission. If clinical suspicion is high, infection should not be ruled out on the basis of RT-PCR alone, and the clinical and epidemiologic situation should be carefully considered. PRIMARY FUNDING SOURCE: National Institute of Allergy and Infectious Diseases, Johns Hopkins Health System, and U.S. Centers for Disease Control and Prevention.


title: Pitfalls of Computed Tomography in the Coronavirus 2019 (COVID-19) Era: A New Perspective on Ground-Glass Opacities

text: Aim To study ground-glass opacities (GGO) not only from the coronavirus 2019 (COVID-19) pneumonia” perspective but also as a radiological presentation of other pathologies with comparable features. Methods We enrolled 33 patients admitted to Policlinico Universitario G. B. Rossi who underwent non-contrast-enhanced (NCE) or contrast-enhanced (CE) chest computed tomography (CT) between March 12 and April 12. All patients with CT-detected ground-glass opacity (GGO) were included. All patients resulted as COVID-19 negative at the reverse transcription-polymerase chain reaction (RT-PCR) assay. We studied the different pathologies underlying GGO features: neoplastic diseases and non-neoplastic diseases (viral pneumonias, interstitial pneumonias, and cardiopulmonary diseases) in order to avoid pitfalls and to reach the correct diagnosis. Results All CT scans detected GGOs. Symptomatic patients were 25/33 (75.7%). At the clinical presentation, they reported fever and dry cough; in six out of 25 cases, dyspnea was also reported (24%). Thirty-three (33; 100%) showed GGO at CT: 15/33 (45.45%) presented pure GGO, and 18/33 (54.54%) showed GGO with consolidation. The RT-PCR assay was negative in 100%. We investigated other potential underlying diseases to explain imaging features: neoplastic causes (8/33, 24.24%) and non-neoplastic causes, in particular, infectious pneumonias (16/33, 48,48 %, viral and fungal), interstitial pneumonias (4/33, 12,12%), and cardio-pulmonary disease (5/33, 15,15%). Conclusions GGO remains a diagnostic challenge. Although CT represents a fundamental diagnostic tool because of its sensitivity, it still needs to be integrated with clinical data to achieve the best clinical management. In the presence of typical imaging features (e.g. GGO and consolidation), the radiologist should focus on the pandemic and manage a suspect patient as COVID-19 positive until proven to be negative.


title: The growth and potential of human antiviral monoclonal antibody therapeutics

text: Monoclonal antibodies (mAbs) have long provided powerful research tools for virologists to understand the mechanisms of virus entry into host cells and of antiviral immunity. Even so, commercial development of human (or humanized) mAbs for the prophylaxis, preemptive and acute treatment of viral infections has been slow. This is surprising, as new antibody discovery tools have increased the speed and precision with which potent neutralizing human antiviral mAbs can be identified. As longstanding barriers to antiviral mAb development, such as antigenic variability of circulating viral strains and the ability of viruses to undergo neutralization escape, are being overcome, deeper insight into the mechanisms of mAb action and engineering of effector functions are also improving the efficacy of antiviral mAbs. These successes, in both industrial and academic laboratories, coupled with ongoing changes in the biomedical and regulatory environments, herald an era when the commercial development of human antiviral mAb therapies will likely surge.


Go back to the basics: Cardiac surgery residents at the time of COVID‐19


title: APA urges HHS to push for partnership to address pandemic

text: Citing the need to provide urgent information on the impact of COVID‐19 on key populations and produce data to target behavioral health interventions to vulnerable populations, the American Psychological Association (APA) is urging the U.S. Department of Health and Human Services (HHS) to establish within the Centers for Disease Control and Prevention a public‐private partnership to address the pandemic.


title: Unraveling the ‘DEAD-box’ helicases of Plasmodium falciparum

text: Abstract The causative agent for the most fatal form of malaria, Plasmodium falciparum, has developed insecticide and drug resistance with time. Therefore combating this disease is becoming increasingly difficult and this calls for finding alternate ways to control malaria. One of the feasible ways could be to find out inhibitors/drugs specific for the indispensable enzymes of malaria parasite such as helicases. These helicases, which contain intrinsic nucleic acid-dependent ATPase activity, are capable of enzymatically unwinding energetically stable duplex nucleic acids into single-stranded templates and are required for all the nucleic acid transactions. Most of the helicases contain a set of nine extremely conserved amino acid sequences, which are called ‘helicase motifs’. Due to the presence of the DEAD (Asp–Glu–Ala–Asp) in one of the conserved motifs, this family is also known as the ‘DEAD-box’ family. In this review, using bioinformatic approach, we describe the ‘DEAD-box’ helicases of malaria parasite P. falciparum. An in depth analysis shows that the parasite contains 22 full-length genes, some of which are homologues of well-characterized helicases of this family from other organisms. Recently we have cloned and characterized the first member of this family, which is a homologue of p68 and is expressed during the schizont stage of the development of the parasite [Pradhan, A., Chauhan, V.S., Tuteja, R., 2005a. A novel ‘DEAD-box’ DNA helicase from Plasmodium falciparum is homologous to p68. Mol. Biochem. Parasitol. 140, 55–60.; Pradhan A., Chauhan V.S., Tuteja R., 2005b. Plasmodium falciparum DNA helicase 60 is a schizont stage specific, bipolar and dual helicase stimulated by PKC phosphorylation. Mol. Biochem. Parasitol. 144, 133–141.]. It will be really interesting to clone and characterize other members of the ‘DEAD-box’ family and understand their role in the replication and transmission of the parasite. These detailed studies may help to identify a parasite-specific enzyme, which could be a potential drug target to treat malaria. The various steps at which this probable drug can act are also discussed.


title: Prevalence of non-influenza respiratory viruses in acute respiratory infection cases in Mexico

text: BACKGROUND: Acute respiratory infections are the leading cause of morbidity and mortality worldwide. Although a viral aetiological agent is estimated to be involved in up to 80% of cases, the majority of these agents have never been specifically identified. Since 2009, diagnostic and surveillance efforts for influenza virus have been applied worldwide. However, insufficient epidemiological information is available for the many other respiratory viruses that can cause Acute respiratory infections. METHODS: This study evaluated the presence of 14 non-influenza respiratory viruses in 872 pharyngeal exudate samples using RT-qPCR. All samples met the operational definition of a probable case of an influenza-like illness or severe acute respiratory infection and had a previous negative result for influenza by RT-qPCR. RESULTS: The presence of at least one non-influenza virus was observed in 312 samples (35.8%). The most frequent viruses were rhinovirus (RV; 33.0%), human respiratory syncytial virus (HRSV; 30.8%) and human metapneumovirus (HMPV; 10.6%). A total of 56 cases of co-infection (17.9%) caused by 2, 3, or 4 viruses were identified. Approximately 62.5% of all positive cases were in children under 9 years of age. CONCLUSION: In this study, we identified 13 non-influenza respiratory viruses that could occur in any season of the year. This study provides evidence for the prevalence and seasonality of a wide range of respiratory viruses that circulate in Mexico and constitute a risk for the population. Additionally, our data suggest that including these tests more widely in the diagnostic algorithm for influenza may reduce the use of unnecessary antibiotics, reduce the hospitalisation time, and enrich national epidemiological data with respect to the infections caused by these viruses.


title: Coronavirus disease (COVID-19): a scoping review

text: BACKGROUND: In December 2019, a pneumonia caused by a novel coronavirus (SARS-CoV-2) emerged in Wuhan, China and has rapidly spread around the world since then. AIM: This study aims to understand the research gaps related to COVID-19 and propose recommendations for future research. METHODS: We undertook a scoping review of COVID-19, comprehensively searching databases and other sources to identify literature on COVID-19 between 1 December 2019 and 6 February 2020. We analysed the sources, publication date, type and topic of the retrieved articles/studies. RESULTS: We included 249 articles in this scoping review. More than half (59.0%) were conducted in China. Guidance/guidelines and consensuses statements (n = 56; 22.5%) were the most common. Most (n = 192; 77.1%) articles were published in peer-reviewed journals, 35 (14.1%) on preprint servers and 22 (8.8%) posted online. Ten genetic studies (4.0%) focused on the origin of SARS-CoV-2 while the topics of molecular studies varied. Nine of 22 epidemiological studies focused on estimating the basic reproduction number of COVID-19 infection (R(0)). Of all identified guidance/guidelines (n = 35), only ten fulfilled the strict principles of evidence-based practice. The number of articles published per day increased rapidly until the end of January. CONCLUSION: The number of articles on COVID-19 steadily increased before 6 February 2020. However, they lack diversity and are almost non-existent in some study fields, such as clinical research. The findings suggest that evidence for the development of clinical practice guidelines and public health policies will be improved when more results from clinical research becomes available.


title: COVID-19 and the Climate Emergency: Do Common Origins and Solutions Reside in the Global Agrifood System?

text: The coronavirus disease 2019 (COVID-19) pandemic and the climate emergency are devastating symptoms of the unsustainability of human society and the decreasing resilience of an unhealthy planet. Here, we discuss whether both COVID-19 and the climate emergency have the same underlying causes, and therefore common solutions, and whether they are rooted in a failing global agrifood system.


title: Molecular modeling, organ culture and reverse genetics for a newly identified human rhinovirus C

text: A recently recognized human rhinovirus species C (HRV-C) is associated with up to half of HRV infections in young children. Here we propagated two HRV-C isolates ex vivo in organ culture of nasal epithelial cells, sequenced a new C15 isolate and developed the first, to our knowledge, reverse genetics system for HRV-C. Using contact points for the known HRV receptors, intercellular adhesion molecule-1 (ICAM-1) and low-density lipoprotein receptor (LDLR), inter- and intraspecies footprint analyses predicted a unique cell attachment site for HRV-Cs. Antibodies directed to binding sites for HRV-A and -B failed to inhibit HRV-C attachment, consistent with the alternative receptor footprint. HRV-A and HRV-B infected HeLa and WisL cells but HRV-C did not. However, HRV-C RNA synthesized in vitro and transfected into both cell types resulted in cytopathic effect and recovery of functional virus, indicating that the viral attachment mechanism is a primary distinguishing feature of HRV-C. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nm.2358) contains supplementary material, which is available to authorized users.


Las enseñanzas del síndrome respiratorio agudo grave


IDENTIFICATION OF RESPIRATORY VIRUSES IN ADULTS: NASOPHARYNGEAL VS OROPHARYNGEAL SAMPLING


title: Ultra-Sensitive High-Resolution Profiling of Anti-SARS-CoV-2 Antibodies for Detecting Early Seroconversion in COVID-19 Patients

text: The COVID-19 pandemic continues to infect millions of people worldwide. In order to curb its spread and reduce morbidity and mortality, it is essential to develop sensitive and quantitative methods that identify infected individuals and enable accurate population-wide screening of both past and present infection. Here we show that Single Molecule Array assays detect seroconversion in COVID-19 patients as soon as one day after symptom onset using less than a microliter of blood. This multiplexed assay format allows us to quantitate IgG, IgM and IgA immunoglobulins against four SARS-CoV-2 targets, thereby interrogating 12 antibody isotype-viral protein interactions to give a high resolution profile of the immune response. Using a cohort of samples collected prior to the outbreak as well as samples collected during the pandemic, we demonstrate a sensitivity of 86% and a specificity of 100% during the first week of infection, and 100% sensitivity and specificity thereafter. This assay should become the gold standard for COVID19 serological profiling and will be a valuable tool for answering important questions about the heterogeneity of clinical presentation seen in the ongoing pandemic.


title: The prevalence of enteric pathogens in diarrhoeic Thoroughbred foals in Britain and Ireland

text: A survey of 77 normal and 326 diarrhoeic foals in Britain and Ireland from 1987 to 1989 revealed a significantly higher prevalence of Group A rotaviruses and Aeromonas hydrophila in diarrhoeic foals. The prevalence of cryptosporidia, potentially pathogenic Escherichia coli, Yersinia enterocolitica and Clostridium perfringens was similar in normal or diarrhoeic foals. Rotaviruses had a similar prevalence in all age groups of scouring foals up to three months of age, with an overall prevalence of 37 per cent among diarrhoeic foals. The number of cases of diarrhoea varied considerably from year to year, but in all three years of the survey rotavirus was a significant pathogen. A comparison of diagnostic tests for rotavirus in the faeces showed electron microscopy (EM) and polyacrylamide gel electrophoresis (PAGE) to have similar sensitivity. The Rotazyme ELISA test kit was found to have the same sensitivity as a combination of EM and PAGE. A. hydrophila had an overall prevalence of 9 per cent among diarrhoeic foals, although its prevalence was higher in some age groups. A. hydrophila has not been established previously as a significant enteric pathogen in foals. Other putative pathogens found at very low prevalence were coronavirus, the putative picobirnavirus, Campylobacter spp. and Salmonella spp. No evidence was found of synergistic effects between rotavirus, cryptosporidia and potentially pathogenic E. coli. Neither coccidia nor non‐Group A rotaviruses were found in any of the samples examined.


title: Extended stability of cyclin D1 contributes to limited cell cycle arrest at G1-phase in BHK-21 cells with Japanese encephalitis virus persistent infection

text: There is increasing evidence that many RNA viruses manipulate cell cycle control to achieve favorable cellular environments for their efficient replication during infection. Although virus-induced G0/G1 arrest often delays early apoptosis temporarily, a prolonged replication of the infected virus leads host cells to eventual death. In contrast, most mammalian cells with RNA virus persistent infection often escape cytolysis in the presence of productive viral replication. In this study, we demonstrated that the extended endurance of cyclin D1 was clearly associated with the suppression of glycogen synthase kinase-3ß (GSK-3ß) expression in BHK-21 cells that are persistently infected with Japanese encephalitis virus (JEV). The G0/G1 arrest of these cells turned much loose compared to the normal BHK-21 cells with JEV acute infection. After cycloheximide treatment, cyclin D1 in the persistently infected cells lasted several hours longer than those in acutely infected cells. Furthermore, both p21(Cip1) and p27(Kip1), positive regulators for cyclin D1 accumulation in the nucleus, were suppressed in their expression, which contrasts with those in JEV acute infection. Inhibition of the GSK-3ß by lithium chloride treatment rescued a significant number of cells from cytolysis in JEV acute infection, which coincided with the levels of cyclin D1 that escaped from proteolysis. Therefore, the limitation of G1/S arrest in the BHK-21 cells with JEV persistent infection is associated with the suppression of GSK-3ß expression, resulting in the extended duration of cyclin D1.


title: Analysis of 2019 novel coronavirus infection and clinical characteristics of outpatients: An epidemiological study from a fever clinic in Wuhan, China

text: BACKGROUND: Since the outbreak of 2019 novel coronavirus (SARS‐CoV‐2) pneumonia, thousands of patients with fever or cough were flocked into fever clinic of designated hospitals in Wuhan, China. To date, no data have ever been reported to reflect the prevalence of Corona Virus Disease 2019 (COVID‐19) among these outpatients. Moreover, it is almost unknown to discriminate COVID‐19 and nucleic acid negative patients based on clinical features in the fever clinics. METHODS: The infectious status of SARS‐CoV‐2 was estimated among the outpatients. The epidemiological and clinical characteristics were compared between COVID‐19 and nucleic acid negative patients. RESULTS: The nucleic acid positive rate for SARS‐CoV‐2 in the outpatients from our fever clinic was 67·1%, while the majority of COVID‐19 patients were mild cases. The predominant initial symptom in those COVID‐19 patients was fever (78.2%), followed by cough (15.6%). Very significantly lower number of eosinophils was characterized in COVID‐19 patients as compared to that of nucleic acid negative patients. More importantly, the proportion of subjects with eosinophil counts lower than normal levels in COVID‐19 patients was much higher than that of nucleic acid negative patients. Fever combined with bilateral ground‐glass opacities in CT imaging and eosinophil count below the normal level are probably a valuable indicator of COVID‐19 infection in those outpatients. CONCLUSIONS: Those findings may provide critical information for the regions such as Europe and United States that are facing the same situation as Wuhan experienced, and could be valuable to prevent those nucleic acid negative patients from misdiagnosis before antibody testing. This article is protected by copyright. All rights reserved.


title: Virus Type and Genomic Load in Acute Bronchiolitis: Severity and Treatment Response With Inhaled Adrenaline

text: Background. Acute bronchiolitis frequently causes infant hospitalization. Studies on different viruses or viral genomic load and disease severity or treatment effect have had conflicting results. We aimed to investigate whether the presence or concentration of individual or multiple viruses were associated with disease severity in acute bronchiolitis and to evaluate whether detected viruses modified the response to inhaled racemic adrenaline. Methods. Nasopharyngeal aspirates were collected from 363 infants with acute bronchiolitis in a randomized, controlled trial that compared inhaled racemic adrenaline versus saline. Virus genome was identified and quantified by polymerase chain reaction analyses. Severity was assessed on the basis of the length of stay and the use of supportive care. Results. Respiratory syncytial virus (83%) and human rhinovirus (34%) were most commonly detected. Seven other viruses were present in 8%–15% of the patients. Two or more viruses (maximum, 7) were detected in 61% of the infants. Virus type or coinfection was not associated with disease severity. A high genomic load of respiratory syncytial virus was associated with a longer length of stay and with an increased frequency of oxygen and ventilatory support use. Treatment effect of inhaled adrenaline was not modified by virus type, load or coinfection. Discussion. In infants hospitalized with acute bronchiolitis, disease severity was not associated with specific viruses or the total number of viruses detected. A high RSV genomic load was associated with more-severe disease. Clinical Trials Registration. NCT00817466 and EudraCT 2009-012667-34.


title: Nanoscale gizmos – the novel fluorescent probes for monitoring protein activity

text: Nanobiotechnology has emerged inherently as an interdisciplinary field, with collaborations from researchers belonging to diverse backgrounds like molecular biology, materials science and organic chemistry. Till the current times, researchers have been able to design numerous types of nanoscale fluorescent tool kits for monitoring protein–protein interactions through real time cellular imagery in a fluorescence microscope. It is apparent that supplementing any protein of interest with a fluorescence habit traces its function and regulation within a cell. Our review therefore highlights the application of several fluorescent probes such as molecular organic dyes, quantum dots (QD) and fluorescent proteins (FPs) to determine activity state, expression and localization of proteins in live and fixed cells. The focus is on Fluorescence Resonance Energy Transfer (FRET) based nanosensors that have been developed by researchers to visualize and monitor protein dynamics and quantify metabolites of diverse nature. FRET based toolkits permit the resolution of ambiguities that arise due to the rotation of sensor molecules and flexibility of the probe. Achievements of live cell imaging and efficient spatiotemporal resolution however have been possible only with the advent of fluorescence microscopic technology, equipped with precisely sensitive automated softwares.


title: Insulin-dependent diabetes mellitus induced in transgenic mice by ectopic expression of class II MHC and interferon-gamma

text: Abstract We have produced transgenic mouse strains harboring class II major histocompatibility complex or interferon-gamma genes linked to the human insulin promoter. These experiments were designed to investigate the consequences of the expression of immunological effector molecules by nonimmunological cells. In both of these studies we observed the disappearance from the pancreas of the insulin-producing β cells coinciding with the development of insulin-dependent diabetes mellitus. Transgenic mice expressing both chains of the I-A gene showed progressive atrophy of the islets of Langerhans, whereas mice expressing interferon-gamma suffered an inflammatory destruction of the islets.


Early Experience in Paris with the Impact of the Covid-19 Pandemic on Vascular Surgery


title: Transport impacts on atmosphere and climate: Aviation

text: Abstract Aviation alters the composition of the atmosphere globally and can thus drive climate change and ozone depletion. The last major international assessment of these impacts was made by the Intergovernmental Panel on Climate Change (IPCC) in 1999. Here, a comprehensive updated assessment of aviation is provided. Scientific advances since the 1999 assessment have reduced key uncertainties, sharpening the quantitative evaluation, yet the basic conclusions remain the same. The climate impact of aviation is driven by long-term impacts from CO2 emissions and shorter-term impacts from non-CO2 emissions and effects, which include the emissions of water vapour, particles and nitrogen oxides (NO x ). The present-day radiative forcing from aviation (2005) is estimated to be 55 mW m−2 (excluding cirrus cloud enhancement), which represents some 3.5% (range 1.3–10%, 90% likelihood range) of current anthropogenic forcing, or 78 mW m−2 including cirrus cloud enhancement, representing 4.9% of current forcing (range 2–14%, 90% likelihood range). According to two SRES-compatible scenarios, future forcings may increase by factors of 3–4 over 2000 levels, in 2050. The effects of aviation emissions of CO2 on global mean surface temperature last for many hundreds of years (in common with other sources), whilst its non-CO2 effects on temperature last for decades. Much progress has been made in the last ten years on characterizing emissions, although major uncertainties remain over the nature of particles. Emissions of NO x result in production of ozone, a climate warming gas, and the reduction of ambient methane (a cooling effect) although the overall balance is warming, based upon current understanding. These NO x emissions from current subsonic aviation do not appear to deplete stratospheric ozone. Despite the progress made on modelling aviation's impacts on tropospheric chemistry, there remains a significant spread in model results. The knowledge of aviation's impacts on cloudiness has also improved: a limited number of studies have demonstrated an increase in cirrus cloud attributable to aviation although the magnitude varies: however, these trend analyses may be impacted by satellite artefacts. The effect of aviation particles on clouds (with and without contrails) may give rise to either a positive forcing or a negative forcing: the modelling and the underlying processes are highly uncertain, although the overall effect of contrails and enhanced cloudiness is considered to be a positive forcing and could be substantial, compared with other effects. The debate over quantification of aviation impacts has also progressed towards studying potential mitigation and the technological and atmospheric tradeoffs. Current studies are still relatively immature and more work is required to determine optimal technological development paths, which is an aspect that atmospheric science has much to contribute. In terms of alternative fuels, liquid hydrogen represents a possibility and may reduce some of aviation's impacts on climate if the fuel is produced in a carbon-neutral way: such fuel is unlikely to be utilized until a ‘hydrogen economy’ develops. The introduction of biofuels as a means of reducing CO2 impacts represents a future possibility. However, even over and above land-use concerns and greenhouse gas budget issues, aviation fuels require strict adherence to safety standards and thus require extra processing compared with biofuels destined for other sectors, where the uptake of such fuel may be more beneficial in the first instance.


title: Publisher Correction: Stabilized coronavirus spikes are resistant to conformational changes induced by receptor recognition or proteolysis

text: A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.


title: Complete Genome Sequence of Porcine Deltacoronavirus Strain CH/Sichuan/S27/2012 from Mainland China

text: We report the first complete genome sequence of porcine deltacoronavirus (PDCoV) strain CH/Sichuan/S27/2012 identified in feces of diarrheic piglets from mainland China in 2012. This strain has two unique in-frame deletions within the ORF1a gene and is phylogenetically between the prototype PDCoV (HKU15) and the 2014 U.S. strains.


title: Ophthalmologic Diseases of Small Mammals

text: Ophthalmic examination of small mammals follows the same principles as in other species. Reference values for tear production and intraocular pressure are provided. Conjunctivitis and epiphora is common in rabbits. Numerous infectious organisms have been isolated from rabbits with conjunctivitis. Dental disease can be a cause for conjunctivitis in rabbits. Treatment consists of topical and oral antimicrobial agents and irrigation of the nasolacrimal system. Underlying dental disease needs to be addressed. Ulcerative keratitis, corneal dystrophy, membranous occlusion of the cornea and eosinophilic keratitis are described. Encephalitozoon cuniculi can cause phacoclastic uveitis in rabbits. A break in the anterior lens capsule exposes lens protein with subsequent anterior uveitis and cataract formation. Cataract surgery can be performed to remove the affected lens. Glaucoma is inherited as an autosomal recessive trait in rabbits. Orbital disease in rabbits may be associated with dental disease. Treatment is often difficult due to the caseous nature of the abscessed material. The most common ophthalmic diseases seen in ferrets include conjunctivitis, corneal endothelial cell degeneration, cataract formation, retinal degeneration and orbital disease. Ophthalmic diseases in guinea pigs include conjunctivitis, dermoids, cataract and osseous metaplasia of the mesectodermal trabecular meshwork. Exophthalmos may be related to dental disease in guinea pigs and chinchillas. Chromodacryorrhea is seen in rats and mice associated with inflammation of the Harderian gland. Sialodacryoadenitis virus is a highly contagious virus that may cause exophthalmos, epiphora and keratoconjunctivitis. Ammonia vapors from soiled bedding can cause keratoconjunctivitis in hamsters.


title: Structural insights into the effect of isonucleosides on B-DNA duplexes using molecular-dynamics simulations

text: Some structural insights into the conformations of the isonucleosides containing duplexes have been provided. Unrestrained molecular-dynamics simulations on 18-mer duplexes with isonucleosides incorporated at the 3'-end or in the center of one strand have been carried out with explicit solvent under periodic boundary conditions using the AMBER force field and the particle mesh Ewald method. The Watson–Crick hydrogen-bonding patterns of the duplexes studied remained intact throughout the simulation. For the modified duplexes, the changes observed in the inter-base pair parameters and backbone torsional angles were primarily localized at the isonucleoside-inserted area. All five structures studied remained in the B-form family. The decreased stacking abilities indicated by the large changes in inter-base pair parameters and the large changes in backbones made the modified duplexes show a minor thermal destabilization in comparison with native DNA. The MM_PBSA method for estimating binding free energies on two complementary strands was used. The results showed that the binding free energies of isonucleoside-incorporated DNA duplexes were lower than the native DNA duplex, which is in good agreement with experimental observations. [Image: see text]


title: Vermindering van diagnostiek en overbehandeling bij RS-virus-bronchiolitis na geprotocolleerde behandeling

text: Purpose: Evaluating the guideline ‘Diagnosis and treatment of respiratory syncytial (RS) virus bronchiolitis’ on the number of chest X-rays, C-reactive proteïn (CRP) counts, leukocyte counts, and antibiotic prescriptions in infants admitted to hospital with RS bronchiolitis. Design: Retrospective ‘before-after’ cohort study. Location: Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands. Patients: Infants admitted with proven RS virus infection. Methods: Guidelines, including sound restriction of performance of X-rays, CRP and leukocytes, were introduced in February 2003. Data from infants admitted with RS virus infection during 1997- 1999 (cohort A) were compared with those admitted from 2003- April 2006 (cohort B) Results: There were 155 infants in cohort A and 170 in cohort B. Implementation of guidelines led to significant reductions of CRP and leukocyte determinations: 49.0% and 48.2%, respectively (both p<0.001) and X-rays: 30.3% (p=0.020). Numbers of antibiotic prescriptions decreased with 55% (p<0.001). The chance of antibiotic prescription increased significantly when X-rays (OR=5.2), CRP (OR=5.4), or leukocytes (OR=4.2) were done. After implementation of the guidelines, the median stay in hospital decreased significantly from 8.0 to 6.0 days (p<0.001; ranges 1-13 days and 2-23 days, respectively). Performing X-ray, CRP or leukocytes, or antibiotic prescription did not significantly alter the total duration of hospital stay. Conclusion: Implementation of the guidelines led to significant decreases in numbers of X-rays, CRP and leukocytes determinations, and antibiotic prescriptions. Our data support the restrictive use of chest X-rays, CRP and leukocyte determinations in infants, admitted to hospital with RS virus bronchiolitis.


title: Pathogenicity and pathogenesis of a United States porcine deltacoronavirus cell culture isolate in 5-day-old neonatal piglets

text: Abstract Porcine deltacoronavirus (PDCoV) was first identified in Hong Kong in 2009–2010 and reported in United States swine for the first time in February 2014. However, diagnostic tools other than polymerase chain reaction for PDCoV detection were lacking and Koch׳s postulates had not been fulfilled to confirm the pathogenic potential of PDCoV. In the present study, PDCoV peptide-specific rabbit antisera were developed and used in immunofluorescence and immunohistochemistry assays to assist PDCoV diagnostics. The pathogenicity and pathogenesis of PDCoV was investigated following orogastric inoculation of 5-day-old piglets with a plaque-purified PDCoV cell culture isolate (3×104 TCID50 per pig). The PDCoV-inoculated piglets developed mild to moderate diarrhea, shed increasing amount of virus in rectal swabs from 2 to 7 days post inoculation, and developed macroscopic and microscopic lesions in small intestines with viral antigen confirmed by immunohistochemistry staining. This study experimentally confirmed PDCoV pathogenicity and characterized PDCoV pathogenesis in neonatal piglets.


title: Is There Still Room for Novel Viral Pathogens in Pediatric Respiratory Tract Infections?

text: Viruses are the most frequent cause of respiratory disease in children. However, despite the advanced diagnostic methods currently in use, in 20 to 50% of respiratory samples a specific pathogen cannot be detected. In this work, we used a metagenomic approach and deep sequencing to examine respiratory samples from children with lower and upper respiratory tract infections that had been previously found negative for 6 bacteria and 15 respiratory viruses by PCR. Nasal washings from 25 children (out of 250) hospitalized with a diagnosis of pneumonia and nasopharyngeal swabs from 46 outpatient children (out of 526) were studied. DNA reads for at least one virus commonly associated to respiratory infections was found in 20 of 25 hospitalized patients, while reads for pathogenic respiratory bacteria were detected in the remaining 5 children. For outpatients, all the samples were pooled into 25 DNA libraries for sequencing. In this case, in 22 of the 25 sequenced libraries at least one respiratory virus was identified, while in all other, but one, pathogenic bacteria were detected. In both patient groups reads for respiratory syncytial virus, coronavirus-OC43, and rhinovirus were identified. In addition, viruses less frequently associated to respiratory infections were also found. Saffold virus was detected in outpatient but not in hospitalized children. Anellovirus, rotavirus, and astrovirus, as well as several animal and plant viruses were detected in both groups. No novel viruses were identified. Adding up the deep sequencing results to the PCR data, 79.2% of 250 hospitalized and 76.6% of 526 ambulatory patients were positive for viruses, and all other children, but one, had pathogenic respiratory bacteria identified. These results suggest that at least in the type of populations studied and with the sampling methods used the odds of finding novel, clinically relevant viruses, in pediatric respiratory infections are low.


title: Where do urologists stand in the era of novel coronavirus-2019 disease

text: The novel coronavirus-2019 disease (COVID-19) pandemic has had devastating consequences on healthcare systems globally. The effect this has on urologists and the patients they care for is not fully understood and presents the challenge of prioritizing the most urgent cases. We aim to review the impact on urology services and evaluate strategies to minimize disruption. RECENT FINDINGS: Various healthcare systems have been forced to postpone treatment for many urological conditions as resources are dedicated to the treatment of COVID-19. Training has been postponed as staff are reallocated to areas of need. Face-to-face contact is largely minimized and innovative, virtual communication methods are used in the outpatient setting and multidisciplinary team meetings. Surgical practice is changing because of the risks posed by COVID-19 and procedures can be prioritized in a nonurgent, low priority, high priority or emergency category. SUMMARY: Although the COVID-19 pandemic will inevitably affect urological services, steps can be taken to mitigate the impact and prioritize the patients most in need of urgent care. Similarly, in future; simulation, e-learning and webinars will allow interaction to share, discuss and debate focused training and education.


Menopausal hormone therapy for women with obesity in the era of COVID-19


title: Preparedness and disinfection of anesthetic equipment in COVID-19

text: • Standards for perioperative supply and equipment precaution for COVID-19 have not been detailed. • Systematically introduces sufficiently preoperative and postoperative equipment precaution; • The sterilization of disassembled anesthesia machine according to high-temperature resistance; • The disinfection of the breathing systems of nondetachable anesthesia machines; • The decontamination of video laryngoscopes, fiberoptic bronchoscopes, and lighted stylets.


title: Mortality statistics in England and Wales: the SARS-CoV-2 paradox

text: OBJECTIVE: To analyse mortality statistics in the United Kingdom during the initial phases of the severe acute respiratory coronavirus 2 (SARS-CoV-2) pandemic and to understand the impact of the pandemic on national mortality. METHODS: Retrospective review of weekly national mortality statistics in the United Kingdom over the past 5 years, including subgroup analysis of respiratory mortality rates. RESULTS: During the early phases of the SARS-CoV-2 pandemic in the first months of 2020, there were consistently fewer deaths per week compared with the preceding 5 years. This pattern was not observed at any other time within the past 5 years. We have termed this phenomenon the “SARS-CoV-2 paradox.” We postulate potential explanations for this seeming paradox and explore the implications of these data. CONCLUSIONS: Paradoxically, but potentially importantly, lower rather than higher weekly mortality rates were observed during the early stages of the SARS-CoV-2 pandemic. This paradox may have implications for current and future healthcare utilisation. A rebound increase in non-SARS-CoV-2 mortality later this year might coincide with the peak of SARS-CoV-2 admissions and mortality.


Smart business networks: Concepts and empirical evidence


title: COVID-19 vaccine design: the Janus face of immune enhancement

text: Previous work on severe acute respiratory syndrome coronavirus (SARS-CoV) vaccines identified cellular immunopathology and antibody-dependent enhancement as potential safety issues. We discuss the implications of these findings for COVID-19 vaccine development and our approach to optimizing for safety and efficacy.


The Parameters to Establish a New Corneal Dystrophy


Exposures among MERS Case-Patients, Saudi Arabia, January–February 2016


title: COVID-19 and congenital heart disease: Results from a nationwide survey

text: Background. The pandemic of Novel Coronavirus Disease 2019 (COVID-19) is challenging, given the large number of hospitalized patients. Cardiovascular co-morbidities are linked to a higher mortality risk. Thus, patients with Congenital Heart Disease (CHD) might represent a high-risk population. Nevertheless, no data about them are available, yet. Hence, we conducted a nationwide survey to assess clinical characteristics and outcomes in patients with congenital heart disease affected by COVID-19. Methods and Results. This is a multi-centre, observational, nationwide survey, involving high-volume Italian CHD centres. COVID-19 diagnosis was defined as either “clinically suspected” or “confirmed”, where a severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) test had been performed and was positive. Cardiovascular comorbidities were observed among adult patients—atrial fibrillation (seven; 9%), hypertension (five; 7%), obesity (seven; 9%) and diabetes (one; 1%)—but were absent among children. Cardiovascular complications were mainly observed in the “confirmed” COVID-19+group, consisting of heart failure (9%), palpitations/arrhythmias (3%), stroke/TIA (3%) and pulmonary hypertension (3%). Cardiovascular symptoms such as chest pain (1%), myocardial injury (1%) and pericardial effusion (1%) were also recorded. On the contrary, CHD patients from the clinically suspected COVID-19 group presented no severe symptoms or complications. Conclusions. Despite previous reports pointing to a higher case-fatality rate among patients with cardiovascular co-morbidities, we observed a mild COVID-19 clinical course in our cohort of CHD patients. Although these results should be confirmed in larger cohorts to investigate the underlying mechanisms, the findings of low cardiovascular complications rates and no deaths are reassuring for CHD patients.


title: Threatening drug‐drug interaction in a kidney transplant patient with Coronavirus Disease 2019 (COVID‐19)

text: During the novel coronavirus pandemic, organ transplant recipients represent a frail susceptible category due to long‐term immunosuppressive therapy. For this reason, clinical manifestations may differ from general population and different treatment approaches may be needed. We present the case of a 36‐year‐old kidney transplanted woman affected by Senior‐Loken syndrome diagnosed with COVID‐19 pneumonia after a contact with her positive mother. Initial symptoms were fatigue, dry cough and coryza; she never had fever nor oxygen supplementation. Hydroxychloroquine and lopinavir/ritonavir were started, and the antiviral drug was replaced with darunavir/cobicistat after two days for diarrhea. Immunosuppressant levels were closely monitored, and we observed very high tacrolimus trough levels despite initial dose reduction. The patient was left with steroid therapy alone. The peculiarity of clinical presentation and the management difficulties represent the flagship of our case‐report. We stress the need for guidelines in transplant recipients with COVID‐19 infection with particular regard to the management of therapy.


title: Impact of severe acute respiratory syndrome on liver transplantation service

text: Severe acute respiratory syndrome (SARS) struck 1755 patients in Hong Kong and developed into a global health crisis. Although the World Health Organization and national health authorities are sparing no effort to contain the disease and to find a cure for the potentially deadly infection, SARS has an impact on our liver transplantation (LTx) program. Before the SARS outbreak, an average of 1 LTx was performed per month in our center. For 6 months since the outbreak, there had been no LTx performed. The intensive care unit had to be dedicated to patients with SARS. Two of the LTx team members were struck by SARS. A survey conducted among LTx recipients and their family members (n = 45) demonstrated symptoms of anxiety and stress in all. Some LTx recipients were treated at the Emergency Department for suspected SARS, which were later confirmed to be false alarms. Many LTx patients were too frightened to come back for follow-up. A new strain of coronavirus was identified as the causative agent. The origin of this virus is uncertain but the probability of zoonoses is being seriously discussed. Not only are immunosuppressed patients exposed to higher risk of infection, but also the waiting list mortality is also expected to increase. The SARS outbreak has demonstrated the vulnerability of an organ transplantation service and reminds us of the fearful possibility of zoonoses in future xeno-transplantation.


title: Chapter 3 Pathogenesis of Viral Infections and Diseases

text: Abstract This chapter describes the pathogenesis of viral infections and diseases, including mechanisms of virulence and disease expression, cellular interactions, and virus-induced neoplasia.