title: Real-time assessment of COVID-19 impact on global surgical case volumes

text: IMPORTANCE: The COVID-19 pandemic has disrupted global surgical capacity. The impact of the pandemic in low and middle income countries has the potential to worsen already strained access to surgical care. Timely assessment of surgical volumes in these countries remains challenging. OBJECTIVE: To determine whether usage data from a globally used anesthesiology calculator mobile application can serve as a proxy for global surgical case volume and contribute to monitoring of the impact of the COVID-19 pandemic, particularly in World Bank low income countries where official data collection is not currently practical. DESIGN: Subset of data from an ongoing observational cohort study of users of the application collected from October 1, 2018 to April 18, 2020. SETTING: The mobile application is available from public sources; users download and use the application per their own clinical needs on personal mobile devices. PARTICIPANTS: No user data was excluded from the study. EXPOSURE(S): Events with impacts on surgical case volumes, including weekends, holidays, and the COVID-19 pandemic. MAIN OUTCOME(S) AND MEASURE(S): It was previously noted that application usage was decreased on weekends and during winter holidays. We subsequently hypothesized that more detailed analysis would reveal impacts of country-specific or region-specific holidays on the volume of app use. RESULTS: 4,300,975 data points from 92,878 unique users were analyzed. Physicians and other anesthesia providers comprised 85.8% of the study population. Application use was reduced on holidays and weekends and correlated with fluctuations in surgical volume. The COVID-19 pandemic was associated with substantial reductions in app use globally and regionally. There was strong cross correlation between COVID-19 case count and reductions in app use. By country, there was a median global reduction in app use to 58% of baseline (interquartile range, 46%−75%). Application use in low-income continues to decline but in high-income countries has stabilized. CONCLUSIONS AND RELEVANCE: Application usage metadata provides a real-time indicator of surgical volume. This data may be used to identify impacted regions where disruptions to surgical care are disproportionate or prolonged. A dashboard for continuous visualization of these data has been deployed.


title: Virus discovery by sequence‐independent genome amplification

text: Genome sequences from several blood borne and respiratory viruses have recently been recovered directly from clinical specimens by variants of a technique known as sequence‐independent single primer amplification. This and related methods are increasingly being used to search for the causes of diseases of presumed infectious aetiology, but for which no agent has yet been found. Other methods that do not require prior knowledge of the genome sequence of any virus that may be present in the patient specimen include whole genome amplification, random PCR and subtractive hybridisation and differential display. This review considers the development and application of these techniques. Copyright © 2006 John Wiley & Sons, Ltd.


Tele-ophthalmology amid COVID-19 pandemic—Hong Kong experience


title: Expression of coronavirus E1 and rotavirus VP10 membrane proteins from synthetic RNA

text: Some viruses acquire their envelopes by budding through internal membranes of their host cell. We have expressed the cloned cDNA for glycoproteins from two such viruses, the E1 protein of coronavirus, which buds in the Golgi region, and VP10 protein of rotavirus, which assembles in the endoplasmic reticulum. Messenger RNA was prepared from both cDNAs by using SP6 polymerase and either translated in vitro or injected into cultured CV1 cells or Xenopus oocytes. In CV1 cells, the El protein was localised to the Golgi region and VP10 protein to the endoplasmic reticulum. In Xenopus oocytes, the E1 protein acquired post‐translational modifications indistinguishable from the sialylated, O‐linked sugars found on viral protein, while the VP10 protein acquired endoglycosidase‐H‐sensitive N‐linked sugars, consistent with their localisation to the Golgi complex and endoplasmic reticulum, respectively. Thus the two proteins provide models with which to study targeting to each of these intracellular compartments. When the RNAs were expressed in matured, meiotic oocytes, the VP10 protein was modified as before, but the E1 protein was processed to a much lesser extent than in interphase oocytes, consistent with a cessation of vesicular transport during cell division.


title: Dose‐dependent Inhibition of Gynecophoral Canal Protein Gene Expression in Vitro in the Schistosome (Schistosoma japonicum) by RNA Interference

text: Abstract The gynecophoral canal protein gene SjGCP of Schistosoma japonicum that is necessary for the pairing between the male and female worms is specifically expressed in the adult male worm. This protein is widely distributed in the adult female worm after pairing. Reverse transcription‐polymerase chain reaction (RT‐PCR) and immunofluorescence were employed to analyze the relationship between the RNAi effect and dsRNA dosage in the parasites. The results revealed that the inhibition of SjGCP expression by siRNA is dose‐dependent. RT‐PCR analysis showed that the SjGCP transcript level was reduced by 75% when 100 nM dsRNA was applied. Edited by: Wang‐Yi LIU


title: Identification of highly conserved regions in L-segment of Crimean–Congo hemorrhagic fever virus and immunoinformatic prediction about potential novel vaccine

text: Crimean–Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic viral disease with a disease fatality rate between 15% and 70%. Despite the wide range of distribution, the virus (CCHFV) is basically endemic in Africa, Asia, eastern Europe, and the Middle East. Acute febrile illness associated with petechiae, disseminated intravascular coagulation, and multiple-organ failure are the main symptoms of the disease. With all these fatal effects, CCHFV is considered a huge threat as no successful therapeutic approach is currently available for the treatment of this disease. In the present study, we have used the immunoinformatics approach to design a potential epitope-based vaccine against the RNA-dependent RNA polymerase-L of CCHFV. Both the T-cell and B-cell epitopes were assessed, and the epitope “DCSSTPPDR” was found to be the most potential one, with 100% conservancy among all the strains of CCHFV. The epitope was also found to interact with both type I and II major histocompatibility complex molecules and is considered nonallergenic as well. In vivo study of our proposed peptide is advised for novel universal vaccine production, which might be an effective path to prevent CCHF disease.


title: Provision of community pharmacy services during COVID-19 pandemic: a cross sectional study of community pharmacists’ experiences with preventative measures and sources of information

text: Background An infectious disease caused by a novel coronavirus (later called COVID-19) reached pandemic levels in 2020 and community pharmacists were involved in responding to this pandemic, also in Kosovo. Objectives To explore the experiences of community pharmacists in relation to provision of community pharmacy services during COVID-19 pandemic. Setting Community pharmacists in Kosovo. Methods This was a cross-sectional study where data was collected via a self-administered online questionnaire, from 264 pharmacists actively practicing in Kosovo during the pandemic. The questionnaire consisted of a combination of closed and open-ended questions, optional statements and statements on a five-point Likert scale, derived at least in part from the Transtheoretical Model. One-way analysis of variance was used to analyze differences in responses to Likert-type items whereas categorical variables were analyzed using Chi square testing. Main outcome measures Community pharmacists’ perceptions on COVID-19 related preventative measures. Results A response rate of 40.6% was achieved. Sufficient and adequate COVID-19-related preventative measures were being implemented by a majority of pharmacies (n = 232; 87.9%), and over two-thirds of respondents agreed/strongly agreed that their pharmacies were sufficiently prepared with protective equipment for their personnel. Implementation of preventative measures was associated with respondents’ perception that pharmacists and the pharmacy profession were valued more by patients during the pandemic and to a lesser degree, by other health professionals. Most commonly stated pros dealt with employee and patient safety, while key cons dealt with increased costs and running out of the necessary protective equipment. Key barriers to pharmacy activities were price increases by wholesalers, and patients’ panic and excessive buying, whereas drivers dealt with professional obligation to assist and opportunity to prove inseparable to other health professionals. The most popular means of accessing COVID-19 related information by pharmacists was via mobile devices and information from professional organizations was considered most useful by pharmacists. Conclusions Community pharmacies actively implemented various measures as precautions to mitigate the spread of COVID-19. Our findings highlight the value of continuous provision of information by professional organizations and use of mobile devices as key means to access information by pharmacists.


title: SARS-CoV-2 disease severity and diabetes: why the connection and what is to be done?

text: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel virus responsible for the current coronavirus disease 2019 (COVID-19) pandemic, has infected over 3.5 million people all over the world since the first case was reported from Wuhan, China 5 months ago. As more epidemiological data regarding COVID-19 patients is acquired, factors that increase the severity of the infection are being identified and reported. One of the most consistent co-morbidities associated with worse outcome in COVID-19 patients is diabetes, along with age and cardiovascular disease. Studies on the association of diabetes with other acute respiratory infections, namely SARS, MERS, and Influenza, outline what seems to be an underlying factor in diabetic patients that makes them more susceptible to complications. In this review we summarize what we think may be the factors driving this pattern between diabetes, aging and poor outcomes in respiratory infections. We also review therapeutic considerations and strategies for treatment of COVID-19 in diabetic patients, and how the additional challenge of this co-morbidity requires attention to glucose homeostasis so as to achieve the best outcomes possible for patients.


title: Development of CRISPR as an Antiviral Strategy to Combat SARS-CoV-2 and Influenza

text: Summary The coronavirus disease 2019 (COVID-19) pandemic, caused by the SARS-CoV-2 virus, has highlighted the need for antiviral approaches that can target emerging viruses with no effective vaccines or pharmaceuticals. Here, we demonstrate a CRISPR-Cas13-based strategy, PAC-MAN (prophylactic antiviral CRISPR in human cells), for viral inhibition that can effectively degrade RNA from SARS-CoV-2 sequences and live influenza A virus (IAV) in human lung epithelial cells. We designed and screened CRISPR RNAs (crRNAs) targeting conserved viral regions and identified functional crRNAs targeting SARS-CoV-2. This approach effectively reduced H1N1 IAV load in respiratory epithelial cells. Our bioinformatic analysis showed that a group of only six crRNAs can target more than 90% of all coronaviruses. With the development of a safe and effective system for respiratory tract delivery, PAC-MAN has the potential to become an important pan-coronavirus inhibition strategy.


title: Clinicopathological Features of Globoid Cell Leucodystrophy in Cats

text: Summary Clinical and pathological findings consistent with globoid cell leucodystrophy (GLD) were evaluated in two domestic shorthaired cats, aged 3 and 4 months. Both showed neurological signs mainly characterized by progressive pelvic limb ataxia, paraplegia with loss of deep pain perception in the pelvic limb, and intentional tremors of the thoracic limbs. Pathological changes affecting the central and peripheral nervous systems were characterised by diffuse, bilateral and symmetrical myelin loss, and marked astrogliosis. In the leucodystrophic areas there was perivascular accumulation of large PAS-positive, non-metachromatic macrophages (globoid cells), with intracytoplasmic accumulation of crystalloid tubular aggregates. Peripheral nerves showed demyelinating features with thin myelin sheaths, myelin splitting, and ballooning; the nerve fibres had bizarre shapes due to the presence of pale inclusions in the Schwann cells. GLD in cats shares clinical and pathological features with the disease described in other animals and human beings. The neurological signs differed from those of other feline inborn neurometabolic diseases and cerebellar hypoplasia.


title: Diversity in cell surface sialic acid presentations: implications for biology and disease

text: Sialic acids (Sias) are typically found as terminal monosaccharides attached to cell surface glycoconjugates. They play many important roles in many physiological and pathological processes, including microbe binding that leads to infections, regulation of the immune response, the progression and spread of human malignancies and in certain aspects of human evolution. This review will provide some examples of these diverse roles of Sias and briefly address immunohistochemical approaches to their detection.


title: Analysis of the intricate effects of polyunsaturated fatty acids and polyphenols on inflammatory pathways in health and disease

text: Prevention and treatment of non-communicable diseases (NCDs), including cardiovascular disease, diabetes, obesity, cancer, Alzheimer’s and Parkinson’s disease, arthritis, non-alcoholic fatty liver disease and various infectious diseases; lately most notably COVID-19 have been in the front line of research worldwide. Although targeting different organs, these pathologies have common biochemical impairments – redox disparity and, prominently, dysregulation of the inflammatory pathways. Research data have shown that diet components like polyphenols, poly-unsaturated fatty acids (PUFAs), fibres as well as lifestyle (fasting, physical exercise) are important factors influencing signalling pathways with a significant potential to improve metabolic homeostasis and immune cells’ functions. In the present manuscript we have reviewed scientific data from recent publications regarding the beneficial cellular and molecular effects induced by dietary plant products, mainly polyphenolic compounds and PUFAs, and summarize the clinical outcomes expected from these types of interventions, in a search for effective long-term approaches to improve the immune system response.


title: Effect of experimental infection of cattle with bovine herpesvirus-1 (BHV-1) on the ex vivo interaction of bovine leukocytes with Mannheimia (Pasteurella) haemolytica leukotoxin

text: Abstract Mannheimia (Pasteurella) haemolytica A1 produces an extracellular leukotoxin (LKT) that is reported to bind the β2-integrin CD11a/CD18 (LEA-1) on ruminant leukocytes. LKT binding induces activation, and subsequent cytolysis, of these cells. It is well known that active viral infection greatly increases the susceptibility of cattle to pasteurellosis. To better understand the mechanism by which this occurs, we investigated the effects of experimental in vivo infection of cattle with bovine herpes virus-1 (BHV-1) on the ex vivo interaction of bovine leukocytes with the M. haemolytica LKT. In this study, we demonstrated that active BHV-1 infection increased the expression of the β2-integrin CD11a/CD18 (as defined by the mAb BAT75) on bovine peripheral blood neutrophils, enhanced the binding of LKT to bronchoalveolar lavage (BAL) leukocytes and peripheral blood neutrophils, and increased the killing of BAL leukocytes and peripheral blood leukocytes by LKT. In addition, BHV-1 greatly increased the number of BAL, resulting in many more LKT-responsive cells being present in the lungs. These findings might explain in part the increased susceptibility of BHV-1 infected cattle to pneumonic pasteurellosis.


title: Inflammation-associated genes: risks and benefits to Foxp3(+) regulatory T-cell function

text: Foxp3(+) regulatory T (Treg) cells prevent the development of autoimmunity and immunopathology, as well as maintaining homeostasis and tolerance to commensal microorganisms. The suppressive activity of Treg cells is their defining characteristic, generating great interest in their therapeutic potential. However, suppressive and effector functions are not entirely exclusive. Considerable evidence points to the ability of supposedly anti-inflammatory Foxp3-expressing Treg cells to also express transcription factors that have been characterized as cardinal drivers of T effector cell function. We will consider the mounting evidence that Treg cells can function in non-suppressive capacities and review the impetus for this functional change, its relevance to developing immune and autoimmune responses and its significance to the development of Treg-based therapies.


title: Chapter 40 Integration of Specialized Research Services into Clinical Laboratory Operations

text: The clinical laboratory can play a useful role in a clinical research program at multiple levels including assistance in developing a laboratory test menu during protocol design, often focused on identifying testing approaches that address specific research questions. Laboratory staff also can provide important input in test performance and test result interpretation in the setting of the specific research questions. In addition, the laboratory can collaborate with investigators in identifying available test methods or developing new testing that will be useful answering specific questions raised in the protocol. Finally, the clinical laboratory can provide important support in translating research questions into new laboratory methods that may not only be useful for the protocol but that may mature into alternative diagnostic or therapeutic applications for patient care. As a model, we outline the approach taken in our laboratory in support of clinical research performed at the National Institutes of Health (NIH) Clinical Center.


title: Five common skin manifestations of COVID‐19 identified

text: COVID‐19, also called coronavirus, is an infectious disease caused by SARS‐CoV‐2. First identified in December 2019, in March 2020 it was announced by the World Health Organisation that COVID‐19 had become a global pandemic. The disease can affect different organ systems, and early reports have suggested skin symptoms. The researchers, working with dermatologists across Spain, reviewed 375 patients with suspected, or confirmed, COVID‐19, with the aim of developing a detailed classification of the different types of skin symptoms associated with COVID‐19 with corresponding images. An initial 120 images were reviewed by four dermatologists, without knowing any other clinical information, and they identified patterns of symptoms associated with COVID‐19. These patterns were then applied to the rest of the images and further refined. Five common skin symptoms were identified this way. These symptoms are chilblain‐like lesions, outbreaks of small blisters, wheals, rashes comprised of small flat and raised red bumps, and livedo and necrosis. Livedo is a skin condition where circulation is impaired within the blood vessels of the skin, necrosis refers to the premature death of skin tissue. These symptoms are associated with different severity of COVID‐19 infection, from less severe disease in the chilblain‐like lesions, to most severe in patients with livedo. The researchers noted that some of the skin symptoms associated with COVID‐19 are common and can have many causes, particularly wheals and rashes. As such, they may not be particularly helpful as an aide to diagnosis.


title: How Has COVID-19 Affected Our Orthopedic Implant Industry Partners? Implications for the Surgeon-Industry Relationship in 2020 and Beyond

text: BACKGROUND: The COVID-19 pandemic has had far-reaching societal and financial consequences. The purpose of this study was to evaluate how COVID-19 has affected AAHKS industry partners and the surgeon-industry relationship, emphasizing education, resource allocation, and strategic direction for the 2nd half of 2020. METHODS: AAHKS industry partners were contacted to participate in a blinded survey and optional interview with the AAHKS Industry Relations Committee. Based on the results, a group of AAHKS member surgeons with disparate practice types were asked to postulate on how the COVID-19 pandemic has and will affect their practice and relationship with Industry. RESULTS: AAHKS industry partner responses indicated decreased resource allocation for regional, “other national,” and AAHKS annual meetings (67%, 55%, and 30%, respectively). Web-based educational content was expected to increase in 2020 and will likely remain a point of emphasis in 2021 (100% and 70% of responders). For Q3/Q4 2020, a significant emphasis was placed on site of service/outpatient TJA and COVID-19-related safety measures (70% and 90% of responders), as well as increased availability of instrumentation and implants (40% and 60%, respectively). CONCLUSION: The COVID-19 pandemic has altered the orthopedic landscape for the foreseeable future. Survey responses by AAHKS industry partners demonstrate a continued commitment to surgeon education with and increasing shift to a web-based platform. Increased resource allocation for outpatient TJA and COVID-19-related safety measures were significant. Articulating optimal mechanisms to aid industry in supporting surgeons with different practice models to meet demand during the second half of fiscal year 2020 will be critical.


title: Comparison of the Biofire FilmArray Respiratory Panel, Seegene AnyplexII RV16, and Argene for the detection of respiratory viruses

text: Abstract Background Respiratory infections are common reasons for hospital admission, and are associated with enormous economic burden due to significant morbidity and mortality. The wide spectrum of microbial agents underlying the pathology renders the diagnosis of respiratory infections challenging. Molecular diagnostics offer an advantage to the current serological and culture-based methods in terms of sensitivity, coverage, hands-on time, and time to results. Objectives This study aimed to compare the clinical performance of three commercial kits for respiratory viral detection. Study design The performance of FilmArray Respiratory Panel, AnyplexII RV16, and Argene was compared using clinical respiratory samples (n = 224, comprising 189 nasopharyngeal swabs in Universal Transport Medium (UTM) and 35 endotracheal aspirates), based on common overlapping targets across the platforms. Influenza A “equivocal” and “no-subtype” samples by FilmArray were further compared to a laboratory-developed Influenza A/B test. Results and Conclusions The overall performance of all three platforms appeared to be comparable with regards to sensitivities (95.8–97.9%) and specificities (96.1–98.0%), detection of coinfections, and distinguishment of influenza from non-influenza cases. “Equivocal” and “no-subtype” samples by FilmArray mostly represented weak Influenza A by laboratory-developed test. Lower respiratory tract samples had comparable final-run success-rates and discordant-rates as compared to UTM. Coronavirus HKU1, which was not targeted by AnyplexII RV16, were detected as OC43. The expected test volume would be the main determinant for the selection of platform. Among the platforms, the FilmArray is the most automated but is of the lowest-throughput and has the highest reagent cost.


title: Surface-enhanced Raman scattering and DFT computational studies of a benzotriazole derivative

text: Here, we report the surface-enhanced Raman scattering (SERS) spectrum of 2-(2′-hydroxy-5′-methylphenyl)benzotriazole (Tinuvin P), a benzotriazole derivative that is the most widely used commercially available UV absorber or stabilizer and is used representatively for the research of photostability mechanism. A full assignment of the Raman spectrum has been made based on the scaled-DFT analysis of the normal vibrational modes. Through the comparative studies on the ordinary Raman spectrum and the SERS spectrum of Tinuvin P, we propose that this molecule binds the Au atom, via the O atom of the hydroxyl or the N1 atom of the benzotriazole moiety, with a perpendicular geometry.


title: Detection of selected pathogens in Apennine wolf (Canis lupus italicus) by a non-invasive GPS-based telemetry sampling of two packs from Majella National Park, Italy

text: In this study, a multi-pathogens survey was conducted to verify the sanitary status of two Italian wolf packs of Majella National Park. Twenty fecal samples (10/pack) were collected using a sampling protocol, based on the combining data from radio-collared wolves with geographic information system (GIS) analysis, allowing to mark off the home range of packs and to recover group-specific and high-quality specimens. Virological screening against the most prevalent canine viruses (protoparvovirus, distemper virus, adenoviruses, and coronaviruses) was carried out by molecular methods, while parasites were detected by means of copromicroscopic and molecular analysis. Canine parvovirus type 2b (CPV-2b) is the most prevalent virus in both packs (7/20), followed by canine adenovirus type 2 (CAdV-2), while no sequences of canine distemper virus and coronaviruses were detected. The sequence analysis of the viruses demonstrated the domestic origin of the infection, highlighting the importance of vaccination of local dogs in order to reduce the risk of exposure of wildlife to these pathogens. Fourteen samples resulted positive for parasites. Capillaria aerophila (sin. Eucoleus aerophilus), Ancylostoma/Uncinaria, Trichuris vulpis eggs, Sarcocystis spp., Cystoisospora canis, and Angiostrongylus vasorum larvae were identified. Echinococcus granulosus sensu stricto (ovine genotype G1) and Giardia duodenalis(canid-specific Assemblage C) were also characterized, providing insights into the wolves’ diet and their effects on environmental contamination. The sampling protocol applied in this study, based on a multidisciplinary approach, represents an innovative tool for the survey of Apennine wolf, able to integrate sanitary data with the ecological and demographic features of this population. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10344-019-1326-y) contains supplementary material, which is available to authorized users.


title: Association of Respiratory Allergy, Asthma and Expression of the SARS-CoV-2 Receptor, ACE2

text: Underlying respiratory allergy and experimental allergen exposure reduce the expression of the SARS-CoV-2 receptor, ACE2, which could lead to reduced COVID-19 susceptibility.


title: Le « syndrome respiratoire du Moyen-Orient » (MERS) : qui est responsable, les chauves-souris ou le dromadaire ?

text: In 2012 a new viral emergent human disease appeared in the Middle East. This entity was named MERS for’ Middle East respiratory syndrome’. By January 9, 2014, the disease had already struck 178 persons of whom 75 died from respiratory failure and diarrhoea. As the new disease was very similar to the deadly SARS (2002–2003) and since it was provoked by a Betacoronavirus, chiroptera were first suspected to be at the origin of this infection. Morever, recent studies performed in Saudi Arabia showed that one individual of the bat Taphozous perforatus harbored a short nucleotide segment identical to the homologous segment present in the viral strain isolated from the index-case of the epidemic. In addition, many strains of Betacoronavirus more or less related to those responsible for the MERS disease in man have been isolated from bats in Africa, Asia and Europe. However, another hypothesis was simultaneously proposed incriminating dromedary (Camelus dromedarius L.) as a likely actor in the transmission to human beings of the disease.We then reviewed data relative to other viral zoonosis in which dromedary was possibly implicated. This led to the provisional conclusion that this large mammal might play a role in the dissemination of the MERS-COV, the etiologic agent of the disease. This is based on epidemiological data and results of several serological surveys in animals.


title: Regulation of hepatitis C virus replication by the core protein through its interaction with viral RNA polymerase

text: Abstract The hepatitis C virus (HCV) core protein is a structural component of the nucleocapsid and has been shown to modulate cellular signaling pathways by interaction with various cellular proteins. In the present study, we investigated the role of HCV core protein in viral RNA replication. Immunoprecipitation experiments demonstrated that the core protein binds to the amino-terminal region of RNA-dependent RNA polymerase (RdRp), which encompasses the finger and palm domains. Direct interaction between HCV RdRp and core protein led to inhibition of RdRp RNA synthesis activity of in vitro. Furthermore, over-expression of core protein, but not its derivatives lacking the RdRp-interacting domain, suppressed HCV replication in a hepatoma cell line harboring an HCV subgenomic replicon RNA. Collectively, our results suggest that the core protein, through binding to RdRp and inhibiting its RNA synthesis activity, is a viral regulator of HCV RNA replication.


title: Longitudinal surveillance of SARS-like coronaviruses in bats by quantitative real-time PCR

text: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available for this article at 10.1007/s12250-015-3703-3 and is accessible for authorized users.


title: Role of cytokines in AIDS wasting

text: Abstract There is now a large literature implicating cytokines in the development of wasting and cachexia commonly observed in a variety of pathophysiologic conditions. In the acquired immunodeficiency syndrome (AIDS), cytokines elicited by primary and secondary infections seem to exert subtle and sustained effects on behavioral, hormonal, and metabolic axes, and their combined effects on appetite and metabolism have been postulated to drive wasting. However, correlations of increased blood levels of a particular cytokine with wasting in AIDS have not been consistent observations, perhaps because cytokines act principally as paracrine and autocrine hormones, as well as indirectly by activating other systems. A better understanding of the mechanisms underlying the catabolic effects of cytokines is clearly needed if more efficacious strategies are to be developed for the prevention and treatment of wasting in AIDS. In this review we first examine the interacting factors contributing to the AIDS wasting syndrome. We then analyze the complex and overlapping role of cytokines in the pathophysiology of this condition, and put forward a number of hypotheses to explain some of the most important features of this syndrome.


title: Lung pathology of severe acute respiratory syndrome (SARS): a study of 8 autopsy cases from Singapore 1 1 This is a US government work. There are no restrictions on its use.

text: Abstract Severe acute respiratory syndrome (SARS) is an infectious condition caused by the SARS-associated coronavirus (SARS-CoV), a new member in the family Coronaviridae. To evaluate the lung pathology in this life-threatening respiratory illness, we studied postmortem lung sections from 8 patients who died from SARS during the spring 2003 Singapore outbreak. The predominant pattern of lung injury in all 8 cases was diffuse alveolar damage. The histology varied according to duration of illness. Cases that were 10 or fewer days in duration demonstrated acute phase diffuse alveolar damage (DAD), airspace edema, and bronchiolar fibrin. Cases with a time course greater than 10 days showed organizing phase DAD, type II pneumocyte hyperplasia, squamous metaplasia, multinucleated giant cells, and acute bronchopneumonia. In acute DAD, pancytokeratin staining was positive in hyaline membranes along alveolar walls and highlighted the absence of pneumocytes. Multinucleated cells were shown to be both type II pneumocytes and macrophages by pancytokeratin, TTF-1, and CD68 staining. SARS-CoV RNA was identified by RT-PCR in 7 of 8 cases in fresh autopsy tissue and in 8 of 8 cases in formalin-fixed, paraffin-embedded lung tissue including the one negative case in fresh tissue. Understanding the pathology of DAD in SARS patients may provide the basis for therapeutic strategies. Further studies of the pathogenesis of SARS may reveal new insights into mechanisms of DAD.


Physicochemical Properties of Cells and Their Effects on Intrinsically Disordered Proteins (IDPs)


The use of in vitro human airway epithelia for the development of novel antivirals


Challenges in interpreting the diagnostic performance of symptoms to predict COVID‐19 status: the case of anosmia


title: Endothelial cells are central orchestrators of cytokine amplification during influenza virus infection

text: Cytokine storm during viral infection is a prospective predictor of morbidity and mortality, yet the cellular sources remain undefined. Here, using genetic and chemical tools to probe functions of the S1P(1) receptor, we elucidate cellular and signaling mechanisms important in initiating cytokine storm. While S1P(1) receptor is expressed on endothelial cells and lymphocytes within lung tissue, S1P(1) agonism suppresses cytokines and innate immune cell recruitment in wild-type and lymphocyte deficient mice, identifying endothelial cells as central regulators of cytokine storm. Furthermore, our data reveal immune cell infiltration and cytokine production as distinct events both orchestrated by endothelial cells. Moreover, we demonstrate that suppression of early innate immune responses through S1P(1) signaling results in reduced mortality during infection with a human pathogenic strain of influenza virus. Modulation of endothelium with a specific agonist suggests that diseases where amplification of cytokine storm is a significant pathological component could be chemically tractable.


The fear of COVID‐19 infection is the main cause of the new diagnoses of hand eczema: report from the frontline in Milan


title: Mathematical prediction in infection

text: Abstract It is now increasingly common for infectious disease epidemics to be analysed with mathematical models. Modelling is possible because epidemics involve relatively simple processes occurring within large populations of individuals. Modelling aims to explain and predict trends in disease incidence, prevalence, morbidity or mortality. Models give important insight into the development of epidemics. Following disease establishment, epidemic growth is approximately exponential. The rate of growth in this phase is primarily determined by the basic reproduction number (R0), the number of secondary cases per primary case when the population is susceptible. R0 also determines the ease with which control policies can control epidemics. Once a significant proportion of the population has been infected, not all contacts of an infected individual will be with susceptible people. Infection can now continue only because new births replenish the susceptible population. Eventually, an endemic equilibrium is reached whereby every infected person infects one other individual on average. Heterogeneity in host susceptibility, infectiousness, human contact patterns and the genetic composition of pathogen populations introduces substantial additional complexity into the models required to model real diseases realistically. The contribution concludes with a brief review of the recent application of mathematical models to emerging human and animal epidemics, notably the spread of HIV in Africa, the variant Creutzfeldt-Jakob disease epidemic in the UK and its relationship to bovine spongiform encephalitis in cattle, the 2001 foot and mouth epidemic in UK livestock, bioterrorism threats such as smallpox, and the SARS epidemics in 2003.


title: Autopsy registry can facilitate COVID‐19 research

text: The autopsy is an important instrument to understand the pathogenesis of diseases, including infectious diseases and novel pathogens like Ebola, SARS or SARS‐CoV‐2 (Mari Saez et al, 2015; Nicholls et al, 2003). The first autopsy studies already suggested important disease mechanisms in fatal COVID‐19 cases with potential therapeutic implications. These include increased thromboembolism and vascular dysfunction (Lax et al, 2020; Menter et al, 2020; Wichmann et al, 2020), infection of endothelial cells (Varga et al, 2020), viral spread in different organs (Puelles et al, 2020), or the pathological mechanisms of lung injury (Ackermann et al, 2020; Schaller et al, 2020). Despite this, no ongoing registry gathered autopsy data, and no specific autopsy registry existed until our recent initiative.


title: Infection rate and clinical management of cancer patients during the COVID-19 pandemic: experience from a tertiary care hospital in northern Italy

text: BACKGROUND: Optimal management of patients with cancer during COVID-19 pandemic is still pending. METHODS: Our patients were advised to maintain their scheduled appointments, and planned cancer treatment was continued without unnecessary delays in an outpatient setting. Additional strict preventive infection measures were rapidly implemented at our outpatient department. When COVID-19 test became widely available, universal testing of healthcare workers and vigorous screening of all patients coming to our facility for COVID-19 infection were performed by SARS-CoV-2 real-time reverse transcription PCR on rhinopharyngeal swab. RESULTS: As of the data cut-off on 9 April 2020, a total of 156 oncology patients with a median age of 67 (range 26–86) years and 63 haematology patients (median age 69 years, range 23–89) were screened for COVID-19 during active cancer treatment. Prevalence (1.8%; 4/219) of COVID-19 in patients with cancer was significantly higher compared with a respective control group of asymptomatic counterparts (p=0.018). Outcomes of COVID-19 positive patients were good, with only one observed death due to progression of advanced metastatic disease. CONCLUSION: Our data indicate that continuation of anticancer treatment in epidemic areas during the COVID-19 pandemic seems to be safe and feasible, if adequate and strict preventive measures are vigorously and successfully carried out.


title: Military-civilian cooperative emergency response to infectious disease prevention and control in China

text: In recent years, the incidence of severe infectious diseases has increased, and the number of emerging infectious diseases continues to increase. The Chinese government and military forces have paid a great deal of attention to infectious disease prevention and control, and using military-civilian cooperation, they have successfully prevented numerous severe epidemic situations, such as severe acute respiratory syndrome (SARS), influenza A (H1N1), avian influenza H5N1 and H7N9, and Ebola hemorrhagic fever, while actively maintained public health, economic development, and national construction. This paper focuses on the mechanisms of the military-cooperative emergency response to infectious diseases--the joint working mechanism, the information-sharing mechanism, the research collaboration mechanism, and the joint disposal mechanism--and presents a sorted summary of the practices and experiences of cooperative emergency responses to infectious diseases. In the future, the Chinese military and the civilian sector will further strengthen the cooperative joint command system and emergency rescue force and will reinforce their collaborative information-sharing platform and technical equipment system to further improve military-civilian collaborative emergency infectious diseases disposal, advance the level of infectious disease prevention and control, and maintain public health.


title: Development and application of a reverse transcription loop-mediated isothermal amplification method for rapid detection of Duck hepatitis A virus type 1

text: We developed and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting Duck hepatitis A virus type 1 (DHAV-1). The amplification could be finished in 1 h under isothermal conditions at 63 °C by employing a set of four primers targeting the 2C gene of DHAV-1. The RT-LAMP assay showed higher sensitivity than the RT-PCR with a detection limit of 0.1 ELD(50) 0.1 ml(−1) of DHAV-1. The RT-LAMP assay was highly specific; no cross-reactivity was observed from the samples of other related viruses, bacteria, allantoic fluid of normal chicken embryos, or the livers of uninfected ducks. Thirty clinical samples were subjected to detection by RT-LAMP, RT-PCR, and virus isolation, which obtained completely consistent, positive results. As a simple, rapid, and accurate detection method, this RT-LAMP assay has important potential applications in the clinical diagnosis of DHAV-1.


title: Experimental Models and Emerging Hypotheses for Acute Lung Injury

text: Acute Lung Injury (ALI) is a complex illness involving the activation of multiple pathways leading to lung injury, resolution and repair. Since the first description of ALI, a great deal of effort has been devoted to exploring the roles of individual pathways in humans and animal models. These have led to a much greater understanding of the complexity of ALI and the links between ALI and systemic multiorgan failure. Despite progress in many areas, we still do not have an integrated understanding of the initiating factors, the key steps in the progression of ALI, and the central processes involved in repair. Although animal models have suggested a range of new hypotheses for study, the major need is for a better understanding of how pathways interact to explain the complexity of the human ALI syndrome and how complementary treatments can be used simultaneously or sequentially to modify the onset, severity and outcome of ALI in humans.


title: Rapid translation of clinical guidelines into executable knowledge: a case study of COVID‐19 and on‐line demonstration

text: The Polyphony programme is a rapidly established collaboration whose aim is to build and maintain a collection of current healthcare knowledge about detection, diagnosis and treatment of COVID‐19 infections, and use Artificial Intelligence (knowledge engineering) techniques to apply the results in patient care. The initial goal is to assess whether the platform is adequate for rapidly building executable models of clinical expertise, while the longer term goal is to use the resulting COVID‐19 knowledge model as a reference and resource for medical training, research and, with partners, develop products and services for better patient care. In this Polyphony progress‐report we describe the first prototype of a care pathway and decision support system that is accessible on OpenClinical.net, a knowledge sharing repository. Pathfinder 1 demonstrates services including situation assessment and inference, decision making, outcome prediction and workflow management. Pathfinder 1 represents encouraging evidence that it is possible to rapidly develop and deploy practical clinical services for patient care and we hope to validate an advanced version in a collaborative internet trial. Finally, we discuss wider implications of the Polyphony framework for developing rapid learning systems in healthcare, and how we may prepare for using AI in future public health emergencies. This article is protected by copyright. All rights reserved.


title: Iron metabolism and lymphocyte characterisation during Covid-19 infection in ICU patients: an observational cohort study

text: BACKGROUND: Iron metabolism and immune response to SARS-CoV-2 have not been described yet in intensive care patients, although they are likely involved in Covid-19 pathogenesis. METHODS: We performed an observational study during the peak of pandemic in our intensive care unit, dosing D-dimer, C-reactive protein, troponin T, lactate dehydrogenase, ferritin, serum iron, transferrin, transferrin saturation, transferrin soluble receptor, lymphocyte count and NK, CD3, CD4, CD8 and B subgroups of 31 patients during the first 2 weeks of their ICU stay. Correlation with mortality and severity at the time of admission was tested with the Spearman coefficient and Mann–Whitney test. Trends over time were tested with the Kruskal–Wallis analysis. RESULTS: Lymphopenia is severe and constant, with a nadir on day 2 of ICU stay (median 0.555 10(9)/L; interquartile range (IQR) 0.450 10(9)/L); all lymphocytic subgroups are dramatically reduced in critically ill patients, while CD4/CD8 ratio remains normal. Neither ferritin nor lymphocyte count follows significant trends in ICU patients. Transferrin saturation is extremely reduced at ICU admission (median 9%; IQR 7%), then significantly increases at days 3 to 6 (median 33%, IQR 26.5%, p value 0.026). The same trend is observed with serum iron levels (median 25.5 μg/L, IQR 69 μg/L at admission; median 73 μg/L, IQR 56 μg/L on days 3 to 6) without reaching statistical significance. Hyperferritinemia is constant during intensive care stay: however, its dosage might be helpful in individuating patients developing haemophagocytic lymphohistiocytosis. D-dimer is elevated and progressively increases from admission (median 1319 μg/L; IQR 1285 μg/L) to days 3 to 6 (median 6820 μg/L; IQR 6619 μg/L), despite not reaching significant results. We describe trends of all the abovementioned parameters during ICU stay. CONCLUSIONS: The description of iron metabolism and lymphocyte count in Covid-19 patients admitted to the intensive care unit provided with this paper might allow a wider understanding of SARS-CoV-2 pathophysiology.


A Joint Exercise against Intentional Biothreats


title: Increased Moraxella and Streptococcus species abundance after severe bronchiolitis is associated with recurrent wheezing

text: Background The role of the airway microbiome in the development of recurrent wheezing and asthma remains uncertain, particularly in the high-risk group of infants hospitalized for bronchiolitis. Objective We sought to examine the relation of the nasal microbiota at bronchiolitis-related hospitalization and 3 later points to the risk of recurrent wheezing by age 3 years. Methods In 17 US centers researchers collected clinical data and nasal swabs from infants hospitalized for bronchiolitis. Trained parents collected nasal swabs 3 weeks after hospitalization and, when healthy, during the summer and 1 year after hospitalization. We applied 16S rRNA gene sequencing to all nasal swabs. We used joint modeling to examine the relation of longitudinal nasal microbiota abundances to the risk of recurrent wheezing. Results Among 842 infants hospitalized for bronchiolitis, there was 88% follow-up at 3 years, and 31% had recurrent wheezing. The median age at enrollment was 3.2 months (interquartile range, 1.7-5.8 months). In joint modeling analyses adjusting for 16 covariates, including viral cause, a 10% increase in relative abundance of Moraxella or Streptococcus species 3 weeks after day 1 of hospitalization was associated with an increased risk of recurrent wheezing (hazard ratio [HR] of 1.38 and 95% high-density interval [HDI] of 1.11-1.85 and HR of 1.76 and 95% HDI of 1.13-3.19, respectively). Increased Streptococcus species abundance the summer after hospitalization was also associated with a greater risk of recurrent wheezing (HR, 1.76; 95% HDI, 1.15-3.27). Conclusions Enrichment of Moraxella or Streptococcus species after bronchiolitis hospitalization was associated with recurrent wheezing by age 3 years, possibly providing new avenues to ameliorate the long-term respiratory outcomes of infants with severe bronchiolitis.


title: Potential of selected Lamiaceae plants in anti(retro)viral therapy

text: Abstract Constant search for new drugs with antiviral properties often extends to products of natural origin. Lamiaceae is one of the most important herbal families, well known for various biological and medicinal effects of a variety of aromatic spices, including thyme, mint, oregano, basil, sage, savory, rosemary, self-heal, hyssop, lemon balm and many others. The paper provides a review of antiviral potential of previously mentioned plants which has been demonstrated so far, with special emphasis on anti-HIV properties. Relevant articles were compiled by searching plant names combined with keywords describing antiviral activity. The antiviral effect is direct, with prominent activity against enveloped viral species. Initial stages of the viral life cycle are the most affected, as these plants appear to be targeting mainly viral structures responsible for attachment to target cells. In case of HIV, there is some activity against key enzymes in the viral life cycle. Even in the case of drug resistance, there is an equal susceptibility to applied herbal preparations. Some in vivo experiments suggest that use of Lamiaceae representatives could help in prevention and treatment of some viral diseases. A possible reduction of side effects of diseases and conventional drug therapy are also some aspects worth further investigations.


title: All-Cause Mortality in Tianjin, China, 1999-2004

text: INTRODUCTION: We analyzed trends of major causes of death in Tianjin, China, from 1999 through 2004 to better inform disease prevention and control programs and policies. METHODS: To report all-cause deaths among Tianjin residents from 1999 through 2004, we standardized mortality rates to the world population in 2000. We analyzed age, sex, and geographic distribution of deaths from different causes and the leading causes of death in Tianjin. RESULTS: The 5 leading causes of death in Tianjin were cardiovascular disease, cerebrovascular disease, malignant neoplasm, chronic lower respiratory disease, and injuries and poisoning. Mortality in Tianjin declined from 0.60% in 1999 to 0.48% in 2004. Noncommunicable diseases accounted for more than 80% of all deaths. Infant and maternal mortality in Tianjin were low. Life expectancy of Tianjin residents increased every year but was consistently longer in women. When deaths from the main chronic diseases are not considered, life expectancy lengthens substantially. CONCLUSION: Chronic diseases are the leading cause of death in Tianjin, China. China should commit additional resources to supporting chronic disease prevention and control programs, including proven special health promotion projects.


title: Enteropathy associated T cell lymphoma in celiac disease: A large retrospective study

text: Abstract Introduction Prognosis of enteropathy-associated T cell lymphoma is poor but predictors of survival remain ill-defined. How clinical presentation, pathological features and therapies influence outcome was evaluated in 37 thoroughly characterized patients with celiac disease and T-cell lymphoma. Patients and methods Medical files were studied retrospectively. Lymphoma and intestinal mucosa were analysed by histopathology, multiplex PCR and intestinal intraepithelial lymphocytes phenotyping. Survival and prognostic factors were analysed using Kaplan–Meier curves with Logrank test and Cox Model. Results Lymphoma complicated non clonal enteropathy, celiac disease (n =15) and type I refractory celiac disease (n =2) in 17 patients and clonal type II refractory celiac disease in 20 patients. Twenty-five patients underwent surgery with resection of the main tumour mass in 22 cases. In univariate analysis, non clonal celiac disease, serum albumin level>21.6g/L at diagnosis, chemotherapy and surgical resection predicted good survival (p =0.0007, p <0.0001, p <0.0001, p <0.0001, respectively). In multivariate analysis, serum albumin level>21.6g/L, chemotherapy and reductive surgery were all significantly associated with increased survival (p <0.002, p <0.03, p <0.03, respectively). Conclusions Our study underlines the prognostic value of celiac disease type in patients with T-cell lymphoma, and suggests that a combination of nutritional, chemotherapy and reductive surgery may improve survival.


title: Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes

text: Rapid, multiplexed, sensitive and specific molecular detection is of great demand in gene profiling, drug screening, clinical diagnostics and environmental analysis(1,2,3). One of the major challenges in multiplexed analysis is to identify each specific reaction with a distinct label or 'code'(4). Two encoding strategies are currently used: positional encoding, in which every potential reaction is preassigned a particular position on a solid-phase support such as a DNA microarray(5,6,7,8), and reaction encoding, where every possible reaction is uniquely tagged with a code that is most often optical or particle based(4,9,10,11,12,13). The micrometer size, polydispersity, complex fabrication process and nonbiocompatibility of current codes limit their usability(1,4,12). Here we demonstrate the synthesis of dendrimer-like DNA-based, fluorescence-intensity-coded nanobarcodes, which contain a built-in code and a probe for molecular recognition. Their application to multiplexed detection of the DNA of several pathogens is first shown using fluorescence microscopy and dot blotting, and further demonstrated using flow cytometry that resulted in detection that was sensitive (attomole) and rapid. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nbt1106) contains supplementary material, which is available to authorized users.


title: Virus characterization and discovery in formalin-fixed paraffin-embedded tissues

text: Abstract Detection and characterization of novel viruses is hampered frequently by the lack of properly stored materials. Especially for the retrospective identification of viruses responsible for past disease outbreaks, often only formalin-fixed paraffin-embedded (FFPE) tissue samples are available. Although FFPE tissues can be used to detect known viral sequences, the application of FFPE tissues for detection of novel viruses is currently unclear. In the present study it was shown that sequence-independent amplification in combination with next-generation sequencing can be used to detect sequences of known and unknown viruses, although with relatively low sensitivity. These findings indicate that this technique could be useful for detecting novel viral sequences in FFPE tissues collected from humans and animals with disease of unknown origin, when other samples are not available. In addition, application of this method to FFPE tissues allows to correlate with the presence of histopathological changes in the corresponding tissue sections.


title: Recommendations on the clinical management of the COVID-19 infection by the «new coronavirus» SARS-CoV2. Spanish Paediatric Association working group

text: Abstract On 31 December 2019, the Wuhan Municipal Committee of Health and Healthcare (Hubei Province, China) reported that there were 27 cases of pneumonia of unknown origin with symptoms starting on the 8 December. There were 7 serious cases with common exposure in market with shellfish, fish, and live animals, in the city of Wuhan. On 7 January 2020, the Chinese authorities identified that the agent causing the outbreak was a new type of virus of the Coronaviridae family, temporarily called «new coronavirus», 2019-nCoV. On January 30th, 2020, the World Health Organisation (WHO) declared the outbreak an International Emergency. On 11 February 2020 the WHO assigned it the name of SARS-CoV2 and COVID-19 (SARS-CoV2 and COVID-19). The Ministry of Health summoned the Specialties Societies to prepare a clinical protocol for the management of COVID-19. The Spanish Paediatric Association appointed a Working Group of the Societies of Paediatric Infectious Diseases and Paediatric Intensive Care to prepare the present recommendations with the evidence available at the time of preparing them.


title: Chapter Eight Hosts and Sources of Endemic Human Coronaviruses

text: Abstract The four endemic human coronaviruses HCoV-229E, -NL63, -OC43, and -HKU1 contribute a considerable share of upper and lower respiratory tract infections in adults and children. While their clinical representation resembles that of many other agents of the common cold, their evolutionary histories, and host associations could provide important insights into the natural history of past human pandemics. For two of these viruses, we have strong evidence suggesting an origin in major livestock species while primordial associations for all four viruses may have existed with bats and rodents. HCoV-NL63 and -229E may originate from bat reservoirs as assumed for many other coronaviruses, but HCoV-OC43 and -HKU1 seem more likely to have speciated from rodent-associated viruses. HCoV-OC43 is thought to have emerged from ancestors in domestic animals such as cattle or swine. The bovine coronavirus has been suggested to be a possible ancestor, from which HCoV-OC43 may have emerged in the context of a pandemic recorded historically at the end of the 19th century. New data suggest that HCoV-229E may actually be transferred from dromedary camels similar to Middle East respiratory syndrome (MERS) coronavirus. This scenario provides important ecological parallels to the present prepandemic pattern of host associations of the MERS coronavirus.


title: Fractal signature of coronaviruses related to severe acute respiratory syndrome

text: PURPOSE: The aim of this work was to investigate the genome of SARS-CoV, MERS-CoV, and SARS-CoV-2 by the paradigm of chaos theory and fractal geometry. Coronavirus is the agent that causes the severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) and emergent infections such as the present pandemic of COVID-19. Understanding its genome pattern is important for developing new and faster ways of testing for identifying the genome of the virus and also for better understanding of its origin and evolution. METHODS: For that, it was calculated the alpha coefficient by detrended fluctuation analysis (DFA) for the sequences of these genomes converted to binary numbers in order to determine if it is a chaotic or a random series of data. Also, it applied the random walking for obtaining a fractal map of the whole genome and calculated the fractal dimension (FD) by box-counting of this map by two different software. RESULTS: With this, it was found that the alpha coefficient of the first SARS viruses was > 0.5, indicating that the series is chaotic or fractal, and has a persistent long-range memory or self-similarity along its sequence. This is not the case for MERS virus, which showed to have a completely random sequence (α < 0.5). For the fractal dimension, SARS viruses presented a FD around 1.5, and for MERS the fractal dimension decreases (FD < 1.5). CONCLUSION: The images generated by random walking of the entire RNA genome are by itself a fractal signature of the virus, which may be applied for studying its origin and for faster diagnostic of COVID-19.


title: A highly conserved WDYPKCDRA epitope in the RNA directed RNA polymerase of human coronaviruses can be used as epitope-based universal vaccine design

text: BACKGROUND: Coronaviruses are the diverse group of RNA virus. From 1960, six strains of human coronaviruses have emerged that includes SARS-CoV and the recent infection by deadly MERS-CoV which is now going to cause another outbreak. Prevention of these viruses is urgent and a universal vaccine for all strain could be a promising solution in this circumstance. In this study we aimed to design an epitope based vaccine against all strain of human coronavirus. RESULTS: Multiple sequence alignment (MSA) approach was employed among spike (S), membrane (M), enveloped (E) and nucleocapsid (N) protein and replicase polyprotein 1ab to identify which one is highly conserve in all coronaviruses strains. Next, we use various in silico tools to predict consensus immunogenic and conserved peptide. We found that conserved region is present only in the RNA directed RNA polymerase protein. In this protein we identified one epitope WDYPKCDRA is highly immunogenic and 100% conserved among all available human coronavirus strains. CONCLUSIONS: Here we suggest in vivo study of our identified novel peptide antigen in RNA directed RNA polymerase protein for universal vaccine – which may be the way to prevent all human coronavirus disease.


title: Synthesis and anti-influenza virus activity of 4-oxo- or thioxo-4,5-dihydrofuro[3,4-c]pyridin-3(1H)-ones

text: Abstract A target-free approach was applied to discover anti-influenza viral compounds, where influenza infected Madin–Darby canine kidney cells were treated 7500 different small organic chemicals individually and reduction of virus-induced cytopathic effect was measured. One of the hit compounds was (Z)-1-((5-fluoro-1H-indol-3-yl)methylene)-6-methyl-4-thioxo-4,5-dihydrofuro[3,4- c ]pyridin-3(1H)-one (15a) with half-maximal effective concentrations of 17.4–21.1μM against influenza A/H1N1, A/H3N2 and B viruses without any cellular toxicity at 900μM. To investigate the structure–activity relationships, two dozens of the hit analogs were synthesized. Among them, 15g, 15j, 15q, 15s, 15t and 15x had anti-influenza viral activity comparable or superior to that of the initial hit. The anti-influenza viral compounds efficiently suppressed not only viral protein level of the infected cells but also production of viral progeny in the culture supernatants in a dose-dependent manner. Based on a mode-of-action study, they did not affect virus entry or RNA replication. Instead, they suppressed viral neuraminidase activity. This study is the first to demonstrate that dihydrofuropyridinones could serve as lead compounds for the discovery of alternative influenza virus inhibitors.


title: Comparison of three rapid influenza diagnostic tests with digital readout systems and one conventional rapid influenza diagnostic test

text: BACKGROUND: Rapid influenza diagnostic tests (RIDTs) show variable sensitivities in clinical settings. We aimed to compare three digital RIDTs and one conventional RIDT. METHODS: We assessed 218 nasopharyngeal swabs from patients between neonates and 90 years old in 2016. Three digital RIDTs were BUDDI, Sofia Influenza A+B Fluorescence Immunoassay, Veritor System Flu A+B assay. One conventional test was the SD Bioline Influenza Ag A/B/A(H1N1/2009). All test results were compared with those from the Anyplex Flu A/B Typing Real‐time Detection real‐time PCR. The four RIDTs were tested with diluted solutions from the National Institute for Biological Standards and Control (NIBSC) to compare lower detection limit. Cross‐reactivity of four RIDTs within other respiratory viruses was identified. RESULTS: For influenza A, BUDDI, Sofia, Veritor, and Bioline showed 87.7%, 94.5%, 87.7%, and 72.6% sensitivity, and 100%, 97.7%, 96.5%, and 100% specificity. For influenza B, BUDDI, Sofia, Veritor, and Bioline showed 81.7%, 91.7%, 81.7%, and 78.3% sensitivity, and 100%, 95.3%, 100%, and 100% specificity, respectively. Each RIDT could detect diluted NIBSC solution, according to the level of dilution and specific influenza subtypes. Cross‐reactivity of four RIDTs with other respiratory viruses was not noted. CONCLUSIONS: Sofia showed the highest sensitivity for influenza A and B detection. BUDDI and Veritor showed higher detection sensitivity than a conventional RIDT for influenza A detection, but similar results for influenza B detection. Further study is needed to compare the test performance of RIDTs according to specific, prevalent influenza subtypes.


title: Glycine 29 Is Critical for Conformational Changes of the Spike Glycoprotein of Mouse Hepatitis Virus A59 Triggered by either Receptor Binding or High pH

text: Mouse hepatitis virus (MHV) uses its N-terminal domain (NTD) of the viral spike (S) protein to bind the host receptor mouse carcinoembryonic antigen-related cell adhesion molecule 1a (mCEACAM1a) and mediate virus entry. Our previous crystal structure study of the MHV NTD/mCEACAM1a complex (G. Peng, D. Sun, K. R. Rajashankar, Z. Qian, et al., Proc Natl Acad Sci U S A 108:10696–10701, 2011, https://doi.org/10.1073/pnas.1104306108) reveals that there are 14 residues in the NTD interacting with the receptor. However, their contribution to receptor binding and virus entry has not been fully investigated. Here we analyzed 13 out of 14 contact residues by mutagenesis and identified I22 as being essential for receptor binding and virus entry. Unexpectedly, we found that G29 was critical for the conformational changes of the S protein triggered by either receptor binding or high pH. Replacement of G29 with A, D, F, K, M, and T, to different extents, caused spontaneous dissociation of S1 from the S protein, resulting in an enhancement of high-pH-triggered receptor-independent syncytium (RIS) formation in HEK293T cells, compared to the wild type (WT). In contrast, replacement of G29 with P, a turn-prone residue with a strict conformation, hindered virus entry and conformational changes of the S protein triggered by either receptor binding or pH 8.0, suggesting that the structural turn around G29 and its flexibility are critical. Finally, stabilization of the NTD by G29P had almost no effect on pH-independent RIS induced by the Y320A mutation in the C-terminal domain (CTD) of the S1 subunit, indicating that there might be an absence of cross talk between the NTD and CTD during conformational changes of the S protein. Our study will aid in better understanding the mechanism of how conformational changes of the S protein are triggered. IMPORTANCE Binding of the MHV S protein to the receptor mCEACAM1a triggers conformational changes of S proteins, leading to the formation of a six-helix bundle and viral and cellular membrane fusion. However, the mechanism by which the conformational change of the S protein is initiated after receptor binding has not been determined. In this study, we showed that while replacement of G29, a residue at the edge of the receptor binding interface and the center of the structural turn after the β1-sheet of the S protein, with D or T triggered spontaneous conformational changes of the S protein and pH-independent RIS, the G29P mutation significantly impeded the conformational changes of S proteins triggered by either receptor binding or pH 8.0. We reason that this structural turn might be critical for conformational changes of the S protein and that altering this structural turn could initiate conformational changes of the S protein, leading to membrane fusion.


title: High host density favors greater virulence: a model of parasite–host dynamics based on multi-type branching processes

text: We use a multitype continuous time Markov branching process model to describe the dynamics of the spread of parasites of two types that can mutate into each other in a common host population. While most mathematical models for the virulence of infectious diseases focus on the interplay between the dynamics of host populations and the optimal characteristics for the success of the pathogen, our model focuses on how pathogen characteristics may change at the start of an epidemic, before the density of susceptible hosts decline. We envisage animal husbandry situations where hosts are at very high density and epidemics are curtailed before host densities are much reduced. The use of three pathogen characteristics: lethality, transmissibility and mutability allows us to investigate the interplay of these in relation to host density. We provide some numerical illustrations and discuss the effects of the size of the enclosure containing the host population on the encounter rate in our model that plays the key role in determining what pathogen type will eventually prevail. We also present a multistage extension of the model to situations where there are several populations and parasites can be transmitted from one of them to another. We conclude that animal husbandry situations with high stock densities will lead to very rapid increases in virulence, where virulent strains are either more transmissible or favoured by mutation. Further the process is affected by the nature of the farm enclosures.


title: A review of the causes of poor fertility in high milk producing dairy cows

text: Abstract Fertility in dairy cows has declined over the past five decades as milk production per cow has increased. Many hypotheses have been proposed to explain this including issues of genetics, physiology, nutrition and management, and these factors have been investigated at the animal, organ and cellular level at critical time points of the productive life of dairy cows. This paper reviews the physiological events and their causes and consequences affecting fertility in dairy cows and summarises these in a downloadable poster. We consider the following points to have the greatest negative impact on fertility and that they need to be prioritised in efforts to ameliorate the problem (others have been included in the review). Firstly, minimise negative energy balance and resolve any infection of the post partum uterus. Secondly, expression and detection of oestrus followed by insemination with high quality semen (day 0). Thirdly, ovulation and fertilisation of a high quality oocyte (day 1). Fourthly, an early increase in progesterone secretion from the corpus luteum (days 3–7). Fifthly, the uterine endometrium must produce an early and appropriate environment to stimulate embryo development (days 6–13). This leads to sixthly, a large embryo producing adequate quantities of interferon tau (days 14–18) that alters uterine prostaglandin secretion and signals maternal recognition of pregnancy (days 16–18). Future strategies to improve dairy cow fertility are needed for the benefit of the dairy industry and for cow welfare and should be based upon an integrative approach of these events.


title: Viral and Bacterial Interactions in the Upper Respiratory Tract

text: Respiratory infectious diseases are mainly caused by viruses or bacteria that often interact with one another. Although their presence is a prerequisite for subsequent infections, viruses and bacteria may be present in the nasopharynx without causing any respiratory symptoms. The upper respiratory tract hosts a vast range of commensals and potential pathogenic bacteria, which form a complex microbial community. This community is assumed to be constantly subject to synergistic and competitive interspecies interactions. Disturbances in the equilibrium, for instance due to the acquisition of new bacteria or viruses, may lead to overgrowth and invasion. A better understanding of the dynamics between commensals and pathogens in the upper respiratory tract may provide better insight into the pathogenesis of respiratory diseases. Here we review the current knowledge regarding specific bacterial–bacterial and viral–bacterial interactions that occur in the upper respiratory niche, and discuss mechanisms by which these interactions might be mediated. Finally, we propose a theoretical model to summarize and illustrate these mechanisms.


Bacterial and viral pathogens in saliva: disease relationship and infectious risk


title: Electrochemical nucleic acid detection based on parallel structural dsDNA/recombinant azurin hybrid

text: Abstract Several challenges remained to fabricate a molecular-level nucleic acid biosensor such as surface immobilization control, single mismatch detection and low current response. To overcome those issues, for the first time, authors presented a novel parallel structural dsDNA/recombinant azurin (PSD/rAzu) hybrid structure for the general nucleic acid detection. The PSD was designed and introduced by the optimized 8 Ag+ ions to have greater conductivity than the canonical dsDNA, and conjugated with rAzu to develop a general platform for electrochemical detection of miRNAs and viral DNAs with high reproducibility and ultra-sensitivity towards single base pair mutation. Thanks to the bifunctional rAzu as the selective spacer and electrochemical signal mediator, in the presence of the target strand, the imperfect PSD switched rapidly to the upright position where the Ag+ ions intercalated between C-C mismatches of dsDNAs at the top of each structure brought further from the electrode surface resulting in a significant electrochemical signal drop of the Ag+ ions. The charge transfer (CT) mechanism across the hybrid structure was simply clarified on the basis of the redox potential location of the species. The electrical conductivity of DNAs were measured using scanning tunneling spectroscopy (STS) at the molecular scale and cyclic voltammetry (CV) technique based on the reduction of Ag+ ion. The proposed PSD/rAzu hybrid structure with a great capability of single mutation recognition and miRNA expression level profiling in cancer cells holds a very promising platform to be studied for further development of various kinds of nanoscale biosensors, bioelectronic devices.


title: Cost-effectiveness analysis of N95 respirators and medical masks to protect healthcare workers in China from respiratory infections

text: BACKGROUND: There are substantial differences between the costs of medical masks and N95 respirators. Cost-effectiveness analysis is required to assist decision-makers evaluating alternative healthcare worker (HCW) mask/respirator strategies. This study aims to compare the cost-effectiveness of N95 respirators and medical masks for protecting HCWs in Beijing, China. METHODS: We developed a cost-effectiveness analysis model utilising efficacy and resource use data from two cluster randomised clinical trials assessing various mask/respirator strategies conducted in HCWs in Level 2 and 3 Beijing hospitals for the 2008–09 and 2009–10 influenza seasons. The main outcome measure was the incremental cost-effectiveness ratio (ICER) per clinical respiratory illness (CRI) case prevented. We used a societal perspective which included intervention costs, the healthcare costs of CRI in HCWs and absenteeism costs. RESULTS: The incremental cost to prevent a CRI case with continuous use of N95 respirators when compared to medical masks ranged from US $490–$1230 (approx. 3000-7600 RMB). One-way sensitivity analysis indicated that the CRI attack rate and intervention effectiveness had the greatest impact on cost-effectiveness. CONCLUSIONS: The determination of cost-effectiveness for mask/respirator strategies will depend on the willingness to pay to prevent a CRI case in a HCW, which will vary between countries. In the case of a highly pathogenic pandemic, respirator use in HCWs would likely be a cost-effective intervention.


title: COVID-19-associated multiple organ damage

text: A 56 years old man presented a particularly severe and multisystemic case of COVID-19. In addition to the common lung and quite common pulmonary embolism and kidney injuries, he presented ocular and intestinal injuries that, to our knowledge, have not been described in COVID-19 patients. Although it is difficult to make pathophysiological hypotheses about a single case, the multiplicity of injured organs argues for a systemic response to pulmonary infection. A better understanding of physiopathology should feed the discussion about therapeutic options in this type of multifocal damage related to SARS-CoV-2.


The Impact of COVID‐19 on IT Services Industry ‐ Expected Transformations


title: Baseline characteristics and clinical symptoms related to respiratory viruses identified among patients presenting with influenza-like illness in primary care

text: Abstract Objectives We aimed to identify patients' clinical characteristics associated with respiratory viruses identified among patients presenting with influenza-like illness (ILI). Methods A sample of patients of all ages presenting with ILI was included by physicians of the French Sentinelles network during two seasons (2015/16 and 2016/17). Nasopharyngeal samples were tested for the presence of influenza virus (IV), respiratory syncytial virus (RSV), human rhinovirus (HRV) and human metapneumovirus (HMPV). Patients' characteristics associated with each of the four virus classes were studied using multivariate logistic regressions. Results A total of 5859 individuals were included in the study: 48.0% tested positive for IV, 7.9% for HRV, 7.5% for RSV and 4.1% for HMPV. Cough was associated with IV (OR 2.14, 95% CI 1.81–2.52) RSV (OR 2.52, 95% CI 1.75–3.74) and HMPV detection (OR 2.15, 95% CI 1.40–3.45). Rhinorrhoea was associated mainly with HRV detection (OR 1.75, 95% CI 1.34–2.32). Headache was associated with IV detection (OR 1.75, 95% CI 1.34–2.32), whereas absence of headache was associated with RSV and HMPV detection. Dyspnoea was associated with RSV detection (OR 2.33, 95% CI 1.73–3.12) and absence of dyspnoea with IV detection. Conjunctivitis was associated with IV detection (OR 1.27, 95% CI 1.08–1.50). Some associations were observed only in children: dyspnoea and cough with RSV detection (age <5 years), conjunctivitis with IV detection (age <15 years). Period of onset of symptoms differed among aetiological diagnoses. Seasonal influenza vaccination decreased the risk of IV detection (OR, 0.67, 95% CI 0.51–0.86). Conclusions This study allowed the identification of symptoms associated with several viral aetiologies in patients with ILI. A proper knowledge and understanding of these clinical signs may improve the medical management of patients.


title: Emerging Infectious Diseases in 2012: 20 Years after the Institute of Medicine Report

text: Twenty years ago (1992), a landmark Institute of Medicine report entitled “Emerging Infections: Microbial Threats to Health in the United States” underscored the important but often underappreciated concept of emerging infectious diseases (EIDs). A review of the progress made and setbacks experienced over the past 2 decades suggests that even though many new diseases have emerged, such as SARS (severe acute respiratory syndrome) and the 2009 pandemic influenza, significant advances have occurred in EID control, prevention, and treatment. Among many elements of the increase in the capacity to control EIDs are genomics-associated advances in microbial detection and treatment, improved disease surveillance, and greater awareness of EIDs and the complicated variables that underlie emergence. In looking back over the past 20 years, it is apparent that we are in a time of great change in which both the challenge of EIDs and our responses to them are being transformed. Recent advances support guarded optimism that further breakthroughs lie ahead.


Critical Need for Implementation of Clinical Pharmacology Principles in Developing Drugs for the Treatment or Prevention of COVID‐19


title: Planning for the Next Global Pandemic

text: Abstract In order to mitigate human and financial losses as a result of future global pandemics, we must plan now. As the Ebola virus pandemic declines, we must reflect on how we have mismanaged this recent international crisis and how we can better prepare for the next global pandemic. Of great concern is the increasing frequency of pandemics occurring over the last few decades. Clearly, the window of opportunity to act is closing. This editorial discusses many issues including priority emerging and re-emerging infectious diseases; the challenges of meeting international health regulations; the strengthening of global health systems; global pandemic funding; and the One Health approach to future pandemic planning. We recommend that the global health community unites to urgently address these issues in order to avoid the next humanitarian crisis.


title: Childhood Rheumatic Diseases and COVID-19 Pandemic: An Intriguing Linkage and a New Horizon

text: As it is known, we are all in a pandemic situation due to a novel coronavirus, officially named “Severe Acute Respiratory Syndrome Coronavirus 2” and the disease caused by the virus named “Coronavirus disease-2019”. The virus seems to has propensity to infect older male individuals with underlying disease. The clinical features were on a large scale that varies from being an asymptomatic carrier to acute respiratory distress syndrome and multiorgan dysfunction. Fever, dry cough and fatigue are the most common symptoms. Not only, the disease seems to be rare and have a milder course in pediatric age but also respiratory failure, multiorgan dysfunction, and death are extremely rare. Although several comorbidities such as hypertension, diabetes and cardiovascular diseases are defined as a risk factor for developing the acute respiratory syndrome and need for intensive care; immune-compromised situations such as rheumatic disease which require immunosuppressive treatment strikingly are not found to be a risk factor for more severe disease course. However, there is a lack of data regarding the effects of “Coronavirus disease-2019” on pediatric patients with rheumatic diseases. Additionally, there are three controversial circumstances that patients with rheumatic diseases are believed to be more likely to have viral infections like “Severe Acute Respiratory Syndrome Coronavirus 2”, on the other hand, antirheumatic drugs may have a protective and therapeutic role in Coronavirus disease-2019 and children are more unlikely to have serious disease course. Therefore, we aimed to have a contributor role for explaining this conundrum and present a bird’s eye view regarding this equivocal issue in this review.


Old Medication for a Novel Disease?


Trainee Led Collaboratives, Clinical Trials And New Opportunities In The Covid‐19 Era


title: The WHO programme for prevention and control of viral, chlamydial, and rickettsial diseases

text: Through the advancement of biological and medical sciences and the application of modern technology, the disease burden imposed by viral, chlamydial and rickettsial disease has steadily decreased. Smallpox has been eradicated, poliomyelitis is under control in many countries, and measles, mumps and rubella viruses may eventually be eliminated in many developed countries. New and improved vaccines have also recently become available for rabies and hepatitis. These are major advancements. Not to be overshadowed however, are the developments which may lead to the prevention or control of other infectious diseases. For many agents, recently acquired knowledge relating to virology, replication, structural and genetic characteristics, and host responses to infection pave the way for disease intervention in numerous ways. For other agents, recent advances in molecular biology make possible new classes of effective vaccines. It is crucial that these advances be incorporated as soon as possible into effective public health programmes for developing as well as developed nations. Much work yet remains, particularly in the prevention and control of respiratory diseases, diarrhoeal diseases, vector-borne diseases and hepatitis. The WHO Viral Diseases Programme has a major role in supporting laboratory and field research on new technologies and intervention strategies, in disseminating technological advances through teaching and training, and in translating the newer knowledge into action programmes for the prevention and control of viral, chlamydial and rickettsial diseases.


title: Health information professionals: delivering core services and value in extraordinary times

text: The 2020 virtual issue of the Health Information and Libraries Journal (HILJ) is published to link to the CILIP Health Libraries Group Conference which was to take place in Scotland 22–25th July. Whilst the conference was postponed in light of the coronavirus (COVID‐19) pandemic, its themes of (i) Working in Partnership; (ii) Resilience and Well‐being; (iii) Public and Patient involvement; (iv) Quality Impact and Metrics; and (v) Improvement and Innovation have nevertheless provided the basis on which to compile this virtual issue. Overarching these themes is a core value of the HIL profession, to provide relevant, timely and sustainable information services, and the articles selected from HILJ (2018 through to March 2020) contribute to the aim of meeting and going beyond these goals under the conference banner of ‘not your average day in the office’. The virtual issue mirrors the format of a regular issue of HILJ, a review article, four original articles and three from our regular features: ‘Dissertations into Practice’, ‘International Perspectives and Initiatives’ and ‘Teaching and Learning in Action’. The authors come from Canada, China, Croatia, Sweden and the UK. All articles included in this issue are available online.


title: A clinicopathological study of three cases of severe acute respiratory syndrome (SARS)

text: Summary Aims The severe acute respiratory syndrome (SARS) caused a large outbreak of atypical pneumonia in Beijing, China from early March 2003. We report the pathological features from three patients who died of SARS. Methods Autopsies were performed on three patients who died 9–15 days after the onset of the illness, and the clinical and laboratory features reviewed. Tissue sections were stained with haematoxylin and eosin (H&E), and in situ reverse transcriptase polymerase chain reaction (RT–PCR) on lung sections was performed using SARS coronavirus-specific primers. Results The typical gross pathological change in the lungs was diffuse haemorrhage on the lung surface. Histopathological examination revealed serous, fibrinous and haemorrhagic inflammation in most pulmonary alveoli, with capillary engorgement and some capillary microthrombosis. The pulmonary alveoli were thickened with interstitial mononuclear inflammatory infiltrates, diffuse alveolar damage, desquamation of pneumocytes and hyaline-membrane formation; fibrinoid material and erythrocytes were present in alveolar spaces. There were thromboemboli in some bronchial arterioles. Haemorrhagic necrosis and reduced numbers of lymphocytes were observed in lymph nodes and spleen. In situ RT–PCR detected SARS coronavirus RNA in type II alveolar cells, interstitial cells and bronchiolar epithelial cells from all three patients Conclusions Severe immunological damage in lung tissue is responsible for the clinical features of SARS.


Evacuation of quarantine-qualified nationals from Wuhan for COVID-19 outbreak – Taiwan experience


title: Comparison of the diagnostic methods on the canine adenovirus type 2 infection

text: Background and aims: Canine adenovirus type 2 (CAV‐2) infection is typically diagnosed histopathologically since intranuclear inclusion bodies (IN/IBs) are demonstrable in the infected lung. However, it is sometimes difficult to identify IN/IBs, particularly in autolyzed tissues or samples from both early and late stages of infection, and other methods were presently developed. Methods: Stray dog samples were evaluated by histopathology, polymerase chain reaction (PCR), and immunohistochemistry (IHC) to investigate the status of the CAV‐2 infection on the stray dogs in Korea. Histologic tests were performed, and dogs with pneumonic lungs were further evaluated by IHC and PCR. Results: Pathognomonic IN/IBs were identified in 3 of 213 lungs; CAV‐2 PCR was positive for 27 of 213 pneumonic lungs. A total of 7 of 27 CAV‐2 PCR‐positive lungs were IHC‐positive. No PCR‐negative lung was IHC‐positive. Positive results were primarily detected in the IN/IBs of the bronchial epithelial cells, macrophages, and very rarely in the cytoplasm of bronchial epithelial cells. Conclusions: IHC was a more reliable diagnostic method than conventional pathologic methods in the present study, and suggests that IHC should be routinely used in the diagnosis of CAV‐2 infection. Further, PCR alone may not be adequate for CAV‐2 diagnosis.


title: Respiratory viral infections are underdiagnosed in patients with suspected sepsis

text: The study aim was to investigate the prevalence and clinical relevance of viral findings by multiplex PCR from the nasopharynx of clinically septic patients during a winter season. During 11 weeks of the influenza epidemic period in January–March 2012, consecutive adult patients suspected to be septic (n = 432) were analyzed with cultures from blood and nasopharynx plus multiplex PCR for respiratory viruses on the nasopharyngeal specimen. The results were compared with those from microbiology analyses ordered as part of standard care. During the winter season, viral respiratory pathogens, mainly influenza A virus, human metapneumovirus, coronavirus, and respiratory syncytial virus were clinically underdiagnosed in 70% of patients positive by the multiplex PCR assay. During the first four weeks of the influenza epidemic, few tests for influenza were ordered by clinicians, indicating low awareness that the epidemic had started. Nasopharyngeal findings of Streptococcus pneumoniae and Haemophilus influenzae by culture correlated to pneumonia diagnosis, and in those patients laboratory signs of viral co-infections were common but rarely suspected by clinicians. The role of respiratory viral infections in patients presenting with a clinical picture of sepsis is underestimated. Specific antiviral treatment might be beneficial in some cases and may reduce spread in a hospital setting. Diagnosing viral infections may promote reduction of unnecessary antibiotic use. It can also be a tool for decisions concerning patient logistics, in order to minimize exposure of susceptible patients and personnel. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10096-017-2990-z) contains supplementary material, which is available to authorized users.


title: N-linked glycosylation and its impact on the electrophoretic mobility and function of the human Proton-Coupled Folate Transporter (HsPCFT)

text: The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. On Western blot, HsPCFT migrates as a broad band (∼55 kDa), higher than predicted (∼50 kDa) in cell lines. Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50°C to avoid aggregation of the protein. Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ∼35 kDa species. Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ∼ 47 kDa protein; substitution of both sites gave a smaller (∼35 kDa) protein. Single mutants retained full transport activity; the double mutant retained a majority of activity. Transport function and molecular size were unchanged when the double-mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains.


title: Co-Circulation of Canine Coronavirus I and IIa/b with High Prevalence and Genetic Diversity in Heilongjiang Province, Northeast China

text: To trace the evolution of canine coronavirus (CCoV), 201 stool samples from diarrheic dogs in northeast China were subjected to reverse transcription-polymerase chain reactions (RT-PCRs) targeting the partial M and S genes of CCoV, followed by an epidemiological analysis. M gene RT-PCRs showed that 28.36% (57/201) of the samples were positive for CCoV; of the 57 positive samples, CCoV-I and CCoV-II accounted for 15.79% (9/57) and 84.21% (48/57), respectively. A sequence comparison of the partial M gene revealed nucleotide homologies of 88.4%–100% among the 57 CCoV strains, and 88.7%–96.2% identity between the 57 CCoV strains and the Chinese reference strain HF3. The CCoV-I and CCoV-II strains exhibited genetic diversity when compared with reference strains from China and other countries. The 57 CCoV strains exhibited high co-infection rates with canine kobuvirus (CaKV) (33.33%) and canine parvovirus-2 (CPV-2) (31.58%). The CCoV prevalence in diarrheic dogs differed significantly with immunization status, regions, seasons, and ages. Moreover, 28 S genes were amplified from the 57 CCoV-positive samples, including 26 CCoV-IIa strains, one CCoV-IIb strain, and one CCoV-I strain. A sequence comparison of the partial S gene revealed 86.3%–100% nucleotide identity among the 26 CCoV-IIa strains, and 89.6%–92.2% identity between the 26 CCoV-IIa strains and the Chinese reference strain V1. The 26 CCoV-IIa strains showed genetic diversity when compared with reference strains from China and other countries. Our data provide evidence that CCoV-I, CCoV-IIa, and CCoV-IIb strains co-circulate in the diarrhoetic dogs in northeast China, high co-infection rates with CaKV and CPV-2 were observed, and the CCoV-II strains exhibited high prevalence and genetic diversity.


title: Impact of confinement housing on study end-points in the calf model of cryptosporidiosis

text: BACKGROUND: Diarrhea is the second leading cause of death in children < 5 years globally and the parasite genus Cryptosporidium is a leading cause of that diarrhea. The global disease burden attributable to cryptosporidiosis is substantial and the only approved chemotherapeutic, nitazoxanide, has poor efficacy in HIV positive children. Chemotherapeutic development is dependent on the calf model of cryptosporidiosis, which is the best approximation of human disease. However, the model is not consistently applied across research studies. Data collection commonly occurs using two different methods: Complete Fecal Collection (CFC), which requires use of confinement housing, and Interval Collection (IC), which permits use of box stalls. CFC mimics human challenge model methodology but it is unknown if confinement housing impacts study end-points and if data gathered via this method is suitable for generalization to human populations. METHODS: Using a modified crossover study design we compared CFC and IC and evaluated the impact of housing on study end-points. At birth, calves were randomly assigned to confinement (n = 14) or box stall housing (n = 9), or were challenged with 5 x 10(7) C. parvum oocysts, and followed for 10 days. Study end-points included fecal oocyst shedding, severity of diarrhea, degree of dehydration, and plasma cortisol. FINDINGS: Calves in confinement had no significant differences in mean log oocysts enumerated per gram of fecal dry matter between CFC and IC samples (P = 0.6), nor were there diurnal variations in oocyst shedding (P = 0.1). Confinement housed calves shed significantly more oocysts (P = 0.05), had higher plasma cortisol (P = 0.001), and required more supportive care (P = 0.0009) than calves in box stalls. CONCLUSION: Housing method confounds study end-points in the calf model of cryptosporidiosis. Due to increased stress data collected from calves in confinement housing may not accurately estimate the efficacy of chemotherapeutics targeting C. parvum.


title: Application of Heteronuclear NMR Spectroscopy to Bioinorganic and Medicinal Chemistry☆

text: Abstract The aim of this article is to provide a comprehensive overview on the use of heteronuclear (in particular, metal) NMR spectroscopy to investigate the behavior of metal ions in biological systems, focusing on some key results, and successful applications of NMR spectroscopy involving direct detection of metals. In this regard, although there is growing interest in the applications of solid-state NMR to biological systems, this subject lies outside the scope of this work, which will be therefore limited to NMR studies carried out in solution. Starting from some review papers published in the last few years, we are here covering relevant results in the field, including the most recent findings reported in the literature to date.


Aux masques citoyens !


Cover Illustration: “Lockdown” mural, Bath Street, Glasgow, by the.rebel.bearUnshackling Plastic Surgery from COVID-19


title: Efficacy of glutathione therapy in relieving dyspnea associated with COVID-19 pneumonia: A report of 2 cases

text: PURPOSE: Infection with COVID-19 potentially can result in severe outcomes and death from “cytokine storm syndrome”, resulting in novel coronavirus pneumonia (NCP) with severe dyspnea, acute respiratory distress syndrome (ARDS), fulminant myocarditis and multiorgan dysfunction with or without disseminated intravascular coagulation. No published treatment to date has been shown to adequately control the inflammation and respiratory symptoms associated with COVID-19, apart from oxygen therapy and assisted ventilation. We evaluated the effects of using high dose oral and/or IV glutathione in the treatment of 2 patients with dyspnea secondary to COVID-19 pneumonia. METHODS: Two patients living in New York City (NYC) with a history of Lyme and tick-borne co-infections experienced a cough and dyspnea and demonstrated radiological findings consistent with novel coronavirus pneumonia (NCP). A trial of 2 g of PO or IV glutathione was used in both patients and improved their dyspnea within 1 h of use. Repeated use of both 2000 mg of PO and IV glutathione was effective in further relieving respiratory symptoms. CONCLUSION: Oral and IV glutathione, glutathione precursors (N-acetyl-cysteine) and alpha lipoic acid may represent a novel treatment approach for blocking NF-κB and addressing “cytokine storm syndrome” and respiratory distress in patients with COVID-19 pneumonia.