title: MicroRNA-34/449 family and viral infections

text: Abstract MicroRNAs are short, endogenous, nonprotein-coding RNAs that are essential for regulation of cellular processes through gene silencing. The miR-34/449 family is conserved in mammalian organisms and generally comprises six homologous genes: miR-34a, miR-34b, miR-34c, miR-449a, miR-449b and miR-449c, at three genomic loci. Strong similarity in the sequence of these miRNAs, particularly at the seed region, predicts robust functional redundancy. A large proportion of the literature on the miR-34/449 family focuses on its role in regulating cell cycle arrest and apoptosis by modulating E2F- and p53-related signaling pathways. A growing subset of the literature reports that the miR-34/449 family is involved in the regulation of immune responses and viral infections, and data suggest the potential for miR-34/446 as a diagnostic and therapeutic target. In this review, we discuss our current understanding of the conservation and transcriptional regulation of the miR-34/449 family and review the literature on its functions in viral infections.


title: Feline leishmaniosis: Is the cat a small dog?

text: Abstract Leishmania infantum is a vector-borne zoonotic disease transmitted by phlebotomine sand flies and dogs are considered the main reservoir of the parasite. Feline leishmaniosis (FeL) caused by L. infantum is an emergent feline disease more and more frequently reported in endemic areas. This review summarizes current knowledge focusing similarities and differences with canine leishmaniosis (CanL). Cats are infected by the same Leishmania species than dogs but prevalence of the infection is lower and cases of disease are less frequently reported. Scarce information is available on adaptive immune response of cats naturally exposed to L. infantum infection and mechanisms responsible for susceptibility or resistance of feline hosts. However, about half of clinical cases of FeL are reported in cats with possible impaired immunocompetence. Coinfections or comorbidities are frequently detected in sick cats and they can contribute to a misrepresentation of clinical FeL albeit lesions associated with the presence of the parasite have been detected in skin, lymph nodes, spleen, bone marrow, liver, oral mucosa, stomach, large bowel, kidney, nasal exudate, lung, eye. As for dogs, skin or mucocutaneous lesions are the most common reason for veterinary consultation and finding on physical examination in cats with leishmaniosis. Molecular investigations of Leishmania DNA and anti- Leishmania antibody detection are largely used with the same methodologies for both CanL and FeL, however few information is available about their diagnostic performance in feline hosts. Treatment of cats with clinical FeL is still empirically based and off label by using the most common drugs prescribed to dogs. Life expectancy of cats with clinical FeL is usually good unless concurrent conditions or complications occur and prognosis does not seem significantly influenced by therapy or retroviral coinfection. According to current knowledge, cats can play a role as additional reservoir host of L. infantum and, in a « One Health » perspective, preventative measures should be taken. In conclusion, albeit feline infection and the associated cat disease caused by L. infantum is increasingly reported in endemic areas and have many similarities with CanL, consolidated evidence-based knowledge is not available and we cannot exclude that important differences between dogs and cats exist about transmission, immunopathogenesis and best practice for management and prevention.


Flattening the Disability Curve: Rehabilitation and Recovery after COVID-19 Infection


title: Coordinated regulation of IFITM1, 2 and 3 genes by an IFN-responsive enhancer through long-range chromatin interactions

text: Abstract Interferon-induced transmembrane protein (IFITM) 1, 2 and 3 genes encode a family of interferon (IFN)-induced transmembrane proteins that block entry of a broad spectrum of pathogens. However, the transcriptional regulation of these genes, especially whether there exist any enhancers and their roles during the IFN induction process remain elusive. Here, through public data mining, episomal luciferase reporter assay and in vivo CRISPR-Cas9 genome editing, we identified an IFN-responsive enhancer located 35kb upstream of IFITM3 gene promoter upregulating the IFN-induced expression of IFITM1, 2 and 3 genes. Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA) and luciferase reporter assay demonstrated that signal transducers and activators of transcription (STAT) 1 bound to the enhancer with the treatment of IFN and was indispensable for the enhancer activity. Furthermore, using chromosome conformation capture technique, we revealed that the IFITM1, 2 and 3 genes physically clustered together and constitutively looped to the distal enhancer through long-range interactions in both HEK293 and A549 cells, providing structural basis for coordinated regulation of IFITM1, 2 and 3 by the enhancer. Finally, we showed that in vivo truncation of the enhancer impaired IFN-induced resistance to influenza A virus (IAV) infection. These findings expand our understanding of the mechanisms underlying the transcriptional regulation of IFITM1, 2 and 3 expression and its ability to mediate IFN signaling.


title: Preparing medical students for a pandemic: a systematic review of student disaster training programmes

text: OBJECTIVE: To identify pandemic and disaster medicine-themed training programmes aimed at medical students and to assess whether these interventions had an effect on objective measures of disaster preparedness and clinical outcomes. To suggest a training approach that can be used to train medical students for the current COVID-19 pandemic. RESULTS: 23 studies met inclusion criteria assessing knowledge (n=18, 78.3%), attitude (n=14, 60.9%) or skill (n=10, 43.5%) following medical student disaster training. No studies assessed clinical improvement. The length of studies ranged from 1 day to 28 days, and the median length of training was 2 days (IQR=1–14). Overall, medical student disaster training programmes improved student disaster and pandemic preparedness and resulted in improved attitude, knowledge and skills. 18 studies used pretest and post-test measures which demonstrated an improvement in all outcomes from all studies. CONCLUSIONS: Implementing disaster training programmes for medical students improves preparedness, knowledge and skills that are important for medical students during times of pandemic. If medical students are recruited to assist in the COVID-19 pandemic, there needs to be a specific training programme for them. This review demonstrates that medical students undergoing appropriate training could play an essential role in pandemic management and suggests a course and assessment structure for medical student COVID-19 training. REGISTRATION: The search strategy was not registered on PROSPERO—the international prospective register of systematic reviews—to prevent unnecessary delay.


title: SARS-CoV-2 is sensitive to type I interferon pretreatment.

text: SARS-CoV-2, a novel coronavirus (CoV), has recently emerged causing an ongoing outbreak of viral pneumonia around the world. While genetically distinct from the original SARS-CoV, both group 2B CoVs share similar genome organization and origins to coronaviruses harbored in bats. Importantly, initial guidance has used insights from SARS-CoV infection to inform treatment and public health strategies. In this report, we evaluate type-I Interferon (IFN-I) sensitivity of SARS-CoV-2 relative to the original SARS-CoV. Our results indicate that while SARS-CoV-2 maintains similar viral replication kinetics to SARS-CoV in Vero cell, the novel CoV is much more sensitive to IFN-I pretreatment. Examining transcriptional factor activation and interferon stimulated gene (ISG) induction, SARS-CoV-2 in the context of type I IFN induces phosphorylation of STAT1 and increased ISG proteins. In contrast, the original SARS-CoV has no evidence for STAT1 phosphorylation or ISG protein increases even in the presence of type I IFN pretreatment. Next, we examined IFN competent Calu3 2B4 cells finding SARS-CoV-2 had reduced viral replication relative to SARS-CoV and induced STAT1 phosphorylation late during infection. Finally, we examined homology between SARS-CoV and SARS-CoV-2 in viral proteins shown to be interferon antagonist. The absence of open reading frame (ORF) 3b and significant changes to ORF6 suggest the two key IFN antagonists may not maintain equivalent function in SARS-CoV-2. Together, the results identify key differences in susceptibility to the IFN-I response between SARS-CoV and SARS-CoV-2. that could help inform disease progression, treatment options, and animal model development.


title: Krisenstrategien der Kliniken während der Pandemie

text: BACKGROUND: The SARS-CoV‑2 pandemic has led worldwide to substantial limitations in healthcare systems. This article describes the recent developments and measures from March through May 2020, which have contributed to the maintenance of ophthalmological care at in-patient departments of ophthalmology. METHODS: PubMed literature search, own data, interhospital survey. RESULTS: The rapid implementation of infection and hygiene control measures and adaptation of standard operating procedures (SOP) to minimize the risk of infection, along with prioritized urgent and emergency care combined with postponement of elective procedures enabled the continuous care of ophthalmological patients. CONCLUSION: Despite the challenge of a significant shift of medical resources during the SARS-CoV‑2 pandemic, medically urgently necessary ophthalmological treatments are continuously provided by maximum care clinics; however, based on currently available data, it cannot be ruled out whether treatment of emergency patients was delayed during the pandemic.


title: Water recycling with PV-powered UV-LED disinfection

text: Abstract Water is in short supply with countries vying for access to river heads. Hong Kong (HK) is no different. It relies on supply from Mainland China with other cities rivalling access. Yet there is still no great impetus for water conservation. This paper reports on progress in setting up a pilot biological wastewater treatment plant and a PV-powered UV-LED disinfection system plus a feasibility study carried out in conjunction with a local developer investigating the application of the treated water for irrigation. The objective is to incorporate the systems within a small-scale community for its contribution to water and energy conservation as well as establish the parameters for replication in other countries with potential up-scaling for urban application.


title: Emerging and re‐emerging viral infections in Europe

text: Emerging viral infections are becoming a serious problem in Europe in the recent years. This is particularly true for severe acute respiratory syndrome (SARS), West Nile virus (WNV) disease, Toscana virus (TOSV) disease, and potentially for avian influenza virus (H5N1). In contrast, emergence or re‐emergence of severe viral infections, including tick borne encephalitis virus, and viral haemorrhagic fever caused by Hantavirus and dengue virus have been frequently reported in several European countries. Laboratory diagnosis of these viral infections based on viral isolation or detection by immune electron microscopy, immunoassay and polymerase chain reaction (PCR) has dramatically improved in the recent years, and SARS represents a good example of a diagnostic approach to emerging viral infections. Finally, old and new promising agents are in the pipeline of pharmaceutical companies to treat emerging viral infections. However only prevention based on large epidemiological studies, and research and development of new vaccines may be able to control and eventually eradicate these deadly viral infections. Copyright © 2006 John Wiley & Sons, Ltd.


title: Efficient Structure Resonance Energy Transfer from Microwaves to Confined Acoustic Vibrations in Viruses

text: Virus is known to resonate in the confined-acoustic dipolar mode with microwave of the same frequency. However this effect was not considered in previous virus-microwave interaction studies and microwave-based virus epidemic prevention. Here we show that this structure-resonant energy transfer effect from microwaves to virus can be efficient enough so that airborne virus was inactivated with reasonable microwave power density safe for the open public. We demonstrate this effect by measuring the residual viral infectivity of influenza A virus after illuminating microwaves with different frequencies and powers. We also established a theoretical model to estimate the microwaves power threshold for virus inactivation and good agreement with experiments was obtained. Such structure-resonant energy transfer induced inactivation is mainly through physically fracturing the virus structure, which was confirmed by real-time reverse transcription polymerase chain reaction. These results provide a pathway toward establishing a new epidemic prevention strategy in open public for airborne virus.


title: Characterisation of the Chemical Composition and Structural Features of Novel Antimicrobial Nanoparticles

text: Three antimicrobial nanoparticle types (AMNP0, AMNP1, and AMNP2) produced using the Tesima(TM) thermal plasma technology were investigated and their compositions were determined using a combination of analytical methods. Scanning electron micrographs provided the morphology of these particles with observed sizes ranging from 10 to 50 nm, whilst FTIR spectra confirmed the absence of polar bonds and organic impurities, and strong Raman active vibrational bands at ca. 1604 and 1311 cm(−1) ascribed to C–C vibrational motions were observed. Carbon signals that resonated at δ(C) 126 ppm in the solid state NMR spectra confirmed that sp(2) hybridised carbons were present in high concentration in two of the nanoparticle types (AMNP1 and AMNP2). X-ray powder diffraction suggested that AMNP0 contains single phase Tungsten carbide (WC) in a high state of purity and multiple phases of WC/WC(1-x) were identified in both AMNP1 and AMNP2. Finally, X-ray photoelectron spectral (XPS) analyses revealed and quantified the elemental ratios in these composite formulations.


title: Adjuvant herbal therapy for targeting susceptibility genes to Kawasaki disease: An overview of epidemiology, pathogenesis, diagnosis and pharmacological treatment of Kawasaki disease

text: Abstract Background Kawasaki disease (KD) is a self-limiting acute systemic vasculitis occur mainly in infants and young children under 5 years old. Although the use of acetylsalicylic acid (AAS) in combination with intravenous immunoglobulin (IVIG) remains the standard therapy to KD, the etiology, genetic susceptibility genes and pathogenic factors of KD are still un-elucidated. Purpose Current obstacles in the treatment of KD include the lack of standard clinical and genetic markers for early diagnosis, possible severe side effect of AAS (Reye's syndrome), and the refractory KD cases with resistance to IVIG therapy, therefore, this review has focused on introducing the current advances in the identification of genetic susceptibility genes, environmental factors, diagnostic markers and adjuvant pharmacological intervention for KD. Results With an overall update in the development of KD from different aspects, our current bioinformatics data has suggested CASP3, CD40 and TLR4 as the possible pathogenic factors or diagnostic markers of KD. Besides, a list of herbal medicines which may work as the adjunct therapy for KD via targeting different proposed molecular targets of KD have also been summarized. Conclusion With the aid of modern pharmacological research and technology, it is anticipated that novel therapeutic remedies, especially active herbal chemicals targeting precise clinical markers of KD could be developed for accurate diagnosis and treatment of the disease.


title: Current awareness: Pharmacoepidemiology and drug safety

text: In order to keep subscribers up‐to‐date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of pharmacoepidemiology and drug safety. Each bibliography is divided into 20 sections: 1 Reviews; 2 General; 3 Anti‐infective Agents; 4 Cardiovascular System Agents; 5 CNS Depressive Agents; 6 Non‐steroidal Anti‐inflammatory Agents; 7 CNS Agents; 8 Anti‐neoplastic Agents; 9 Haematological Agents; 10 Neuroregulator‐Blocking Agents; 11 Dermatological Agents; 12 Immunosuppressive Agents; 13 Autonomic Agents; 14 Respiratory System Agents; 15 Neuromuscular Agents; 16 Reproductive System Agents; 17 Gastrointestinal System Agents; 18 Anti‐inflammatory Agents ‐ Steroidal; 19 Teratogens/fetal exposure; 20 Others. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted.


title: Can the Coronavirus Disease 2019 (COVID-19) Affect the Eyes? A Review of Coronaviruses and Ocular Implications in Humans and Animals

text: In December 2019, a novel coronavirus (CoV) epidemic, caused by the severe acute respiratory syndrome coronavirus – 2 (SARS-CoV-2) emerged from China. This virus causes the coronavirus disease 2019 (COVID-19). Since then, there have been anecdotal reports of ocular infection. The ocular implications of human CoV infections have not been widely studied. However, CoVs have been known to cause various ocular infections in animals. Clinical entities such as conjunctivitis, anterior uveitis, retinitis, and optic neuritis have been documented in feline and murine models. In this article, the current evidence suggesting possible human CoV infection of ocular tissue is reviewed. The review article will also highlight animal CoVs and their associated ocular infections. We hope that this article will serve as a start for further research into the ocular implications of human CoV infections.


title: The impact of the COVID‐19 pandemic on eating disorder risk and symptoms

text: The current COVID‐19 pandemic has created a global context likely to increase eating disorder (ED) risk and symptoms, decrease factors that protect against EDs, and exacerbate barriers to care. Three pathways exist by which this pandemic may exacerbate ED risk. One, the disruptions to daily routines and constraints to outdoor activities may increase weight and shape concerns, and negatively impact eating, exercise, and sleeping patterns, which may in turn increase ED risk and symptoms. Relatedly, the pandemic and accompanying social restrictions may deprive individuals of social support and adaptive coping strategies, thereby potentially elevating ED risk and symptoms by removing protective factors. Two, increased exposure to ED‐specific or anxiety‐provoking media, as well as increased reliance on video conferencing, may increase ED risk and symptoms. Three, fears of contagion may increase ED symptoms specifically related to health concerns, or by the pursuit of restrictive diets focused on increasing immunity. In addition, elevated rates of stress and negative affect due to the pandemic and social isolation may also contribute to increasing risk. Evaluating and assessing these factors are key to better understanding the impact of the pandemic on ED risk and recovery and to inform resource dissemination and targets.


title: Molecular characterization of Cryptosporidium isolates from diarrheal dairy calves in France

text: Abstract Cryptosporidium is an obligate intracellular protist parasite infecting a wide range of vertebrate hosts and causes significant intestinal disease in both animals and humans, as some species are zoonotic. Cattle and especially calves have been identified as one of the most common reservoirs of this protist. However, little is known about the genetics of Cryptosporidium in calves in some regions of France. The aim of this study was to detect and isolate Cryptosporidium spp. in faecal samples from naturally infected pre-weaned calves (≤45 days-old) in France. A total of 35 diarrhoeic pre-weaned calf faecal samples were collected from 26 dairy cattle farms in six departments (French administrative provinces). Cryptosporidium presence was established by microscopically screening samples for oocystes with an immunofluorescent (DFA) staining method. DFA-positive samples were then analysed by PCR-RFLP and 18S rRNA gene sequencing to determine species. Cryptosporidium parvum-positive samples were subtyped via nested PCR analysis of a partial fragment of the 60 kDa glycoprotein (gp60) gene product. Data were then integrated into phylogenetic tree analysis. DFA revealed the presence of Cryptosporidium oocysts in 31 out of 35 (88%) samples. Combined with 18S rRNA gene analysis results, C. parvum was detected in 30 samples. Subtyping analysis in 27/30 samples (90%) of the C. parvum isolates revealed two zoonotic subtype families, IIa (24/27) and IId (3/27). Four subtypes were recognised within the subtype family IIa, including the hypertransmissible IIaA15G2R1 subtype that is the most frequently reported worldwide (21/27), IIaA17G3R1 (1/27), IIaA17G1R1 (1/27), and IIaA19G1R1 (1/27). Two subtypes were recognised within the IId subtype family including IIdA22G1 (2/27) and IIdA27G1 (1/27). These findings illustrate the high occurrence of Cryptosporidium in calves in dairy herds and increase the diversity of molecularly characterised C. parvum isolates with the first description of IIaA17G3R1, IIaA19G1R1, and IId subtypes in France. The presence of zoonotic C. parvum subtype families (IIa, IId) in this study suggests that pre-weaned calves are likely to be a significant reservoir of zoonotic C. parvum, and highlights the importance of animal to human cryptosporidiosis transmission risk. Further molecular studies in calves and small ruminants from other French regions are required to better understand the epidemiology of cryptosporidiosis in France.


Infection control and MERS-CoV in health-care workers


title: CID‐19 pulmonary infection in erythrodermic psoriatic patient with oligodendroglioma: safety and compatibility of apremilast with critical intensive care management

text: Novel coronavirus 2019 (SARS-CoV2) pandemic has particularly affected Italy, with a profound impact on the therapeutic strategy for complex disorder such as psoriasis, whose extensive skin damage might expose to an increased infective risk compared to the general population. Psoriasis treatment relies on immunosuppression, and although most experts agree that the benefit-to risk-ratio is in favor of maintaining selective biologic therapies, and small molecules such as apremilast, they recommend dismission if severe COVID-19 symptoms occur.


title: 2448. Clinical Presentation and Outcomes of Long-Term Care Residents with Coronavirus Respiratory Infection: A Retrospective Cohort Study

text: BACKGROUND: Human coronaviruses (CoVs) are a major cause of respiratory infection and institutional outbreaks, yet the epidemiology and clinical outcomes of these viruses is poorly described among the elderly residing in long-term care facilities (LTCFs). METHODS: We performed a retrospective cohort study of LTCF residents with positive nasopharyngeal or mid-turbinate swabs for CoVs (OC43, 229E, NL63 and HKU1) between January 2013 and December 2018. Demographic and clinical data were obtained from resident charts including clinical presentation, treatment, outcome, and transmission to other residents. Variables were compared using univariate analysis. RESULTS: 3268 residents met inclusion criteria (median age 93 years, 90% male) comprising 7.5% (246/3268) of all positive respiratory virus specimens detected during the study period. 97(39%) of cases were associated with a respiratory outbreak while 149(61%) were sporadic cases that did not result in transmission. OC43 (52%) was the most commonly identified CoV and was more commonly associated with outbreak cases (76% vs. 37%; P < 0.001). In total, 87% of all cases had two or more of runny nose/congestion, cough, sore throat/hoarse voice or fever. The most common symptoms among residents were cough (85%), runny nose/congestion (79%), and sore throat/hoarse voice (59%) and only 17% of residents had a measured temperature of ≥ 37.8C. Only 6% of residents received antibiotic treatment for suspected secondary bacterial pneumonia. The 30-day mortality rate was 3.7% with 67% of deaths attributable to the CoV infection. There was no statistically significant difference in symptoms, treatment or outcomes associated with outbreaks or seasonality. CONCLUSION: CoVs make up an important proportion of respiratory viral infections among LTCF residents and may result in frequent outbreaks. Most residents remain afebrile and have self-limited illness while only a small minority develop secondary bacterial pneumonia and death. Given these findings the benefits of control measures should be weighed against the impact on resident quality of life. DISCLOSURES: All authors: No reported disclosures.


title: Effect of the COVID‐19 pandemic on anxiety among children with cystic fibrosis and their mothers

text: BACKGROUND: We aimed to evaluate anxiety among children with cystic fibrosis (CF) and their mothers related to the COVID‐19 pandemic. METHODS: A total of 45 patients with CF and their mothers were enrolled in the study together with 90 age‐matched healthy children and their mothers as a control group. The State and Trait Anxiety Inventory (STAI) was administered by teleconference with children aged 13–18 years old and their mothers. The STAI for children was administered with children aged 9–12 years. Results were compared with age‐matched healthy children and their mothers. The relationship between anxiety scores of children with CF and their mothers was evaluated by comparing with clinical data of children with CF. At the conclusion of the teleconference, mothers were asked whether their anxiety had changed as a result of the interview. RESULTS: It was found that healthy children aged 13–18 years had higher state anxiety scores than age‐matched children with CF. Mothers of children with CF had higher trait anxiety scores, especially those of children aged 0–12 years, than mothers of healthy children (p<0.05). For mothers of children with CF, state anxiety scores were higher among those whose children had chronic Pseudomonas infection (p<0.05). Most mothers of children with CF stated that their anxiety decreased following the interview. CONCLUSION: The COVID‐19 pandemic may increase anxiety among mothers of children with CF as well those with healthy children. However, COVID‐19 had no effect on the anxiety of children with CF. Informing parents of children with CF about COVID‐19 by teleconference may decrease anxiety. This article is protected by copyright. All rights reserved.


title: An evidence-based framework for priority clinical research questions for COVID-19

text: BACKGROUND: On 31 December, 2019, the World Health Organization China Country Office was informed of cases of pneumonia of unknown aetiology. Since then, there have been over 75 000 cases globally of the 2019 novel coronavirus (COVID-19), 2000 deaths, and over 14 000 cases recovered. Outbreaks of novel agents represent opportunities for clinical research to inform real-time public health action. In 2018, we conducted a systematic review to identify priority research questions for Severe Acute Respiratory Syndrome-related coronavirus (SARS-CoV) and Middle East Respiratory Syndrome-related coronavirus (MERS-CoV). Here, we review information available on COVID-19 and provide an evidenced-based framework for priority clinical research in the current outbreak. METHODS: Three bibliographic databases were searched to identify clinical studies published on SARS-CoV and MERS-CoV in the outbreak setting. Studies were grouped thematically according to clinical research questions addressed. In February 2020, available information on COVID19 was reviewed and compared to the results of the SARS-CoV and MERS-CoV systematic review. RESULTS: From the research objectives for SARS-CoV and MERS-CoV, ten themes in the literature were identified: Clinical characterisation, prognosis, diagnosis, clinical management, viral pathogenesis, epidemiological characterisation, infection prevention and control/transmission, susceptibility, psychosocial, and aetiology. For COVID19, some information on clinical presentation, diagnostic testing, and aetiology is available but many clinical research gaps have yet to be filled. CONCLUSIONS: Based on a systematic review of other severe coronaviruses, we summarise the state of clinical research for COVID-19, highlight the research gaps, and provide recommendations for the implementation of standardised protocols. Data based on internationally standardised protocols will inform clinical practice real-time.


title: How Pathogen-Derived Cysteine Proteases Modulate Host Immune Responses

text: In mammals, cysteine proteases are essential for the induction and development of both innate and adaptive immune responses. These proteases play a role in antigen-and pathogen-recognition and elimination, signal processing and cell homeostasis. Many pathogens also secrete cysteine proteases that often act on the same target proteins as the mammalian proteases and thereby can modulate host immunity from initial recognition to effector mechanisms. Pathogen-derived proteases range from nonspecific proteases that degrade multiple proteins involved in the immune response to enzymes that are very specific in their mode of action. Here, we overview current knowledge of pathogen-derived cysteine proteases that modulate immune responses by altering the normal function of key receptors or pathways in the mammalian immune system.


title: An update on COVID-19 for the otorhinolaryngologist – a Brazilian Association of Otolaryngology and Cervicofacial Surgery (ABORL-CCF) Position Statement

text: Abstract Introduction We are facing a pandemic with a great impact worldwide, as a result of the rapid spread of the novel coronavirus (COVID-19). The medical community is still getting to know behavior of this virus and the consequences from a population point of view. All this knowledge is extremely dynamic, so some behaviors are still not well established. Otorhinolaryngologists have a central role in the management of this situation, in which they must assess the patient, avoid contamination to and by health professionals and other patients. Thus, the recommendations of the Brazilian Association of Otorhinolaryngology and Cervical-Facial Surgery (ABORL-CCF) have the main objective of reducing the spread of the new coronavirus during otorhinolaryngological care and assisting in the management of these patients. Methods Review of the main recommendations of national and international scientific societies, decisions by government agencies and class councils. The topics will be related to the general aspects of COVID-19, personal protective equipment, care in patient assistance, endoscopic exam routines and the management of sinonasal, otological and pediatric evaluations related to COVID-19. Results The use of personal protective equipment is considered crucial in routine ENT care. We recommend postponing appointments, exams and elective surgeries to reduce the spread of COVID-19. Similarly, we recommend changing routines in several areas of otolaryngology. Additionally, guidance is provided on the use of telemedicine resources during the pandemic period. Conclusions We are still at the beginning of the COVID-19 pandemic and scientific evidence is still scarce and incomplete, so these ABORL-CCF recommendations for otorhinolaryngologists may be updated based on new knowledge and the pattern of the new coronavirus spread.


title: A plastic surgery service response to COVID-19 in one of the largest teaching hospitals in Europe

text: Summary COVID-19 is presenting a colossal challenge to frontline NHS staff. This paper highlights how plastic surgery teams can use their diverse skills and resources in times of crisis. Through effective strategy and leadership we present how we are adapting as a department to serve our plastic surgery patients, other hospital teams and the Trust.


title: A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys

text: Bats are source of coronaviruses closely related to the severe acute respiratory syndrome (SARS) virus. Numerous studies have been carried out to identify new bat viruses related to SARS-coronavirus (bat-SARS-like CoVs) using a reverse-transcribed-polymerase chain reaction assay. However, a qualitative PCR could underestimate the prevalence of infection, affecting the epidemiological evaluation of bats in viral ecology. In this work an SYBR Green-real time PCR assay was developed for diagnosing infection with SARS-related coronaviruses from bat guano and was applied as screening tool in a survey carried out on 45 greater horseshoe bats (Rhinolophus ferrumequinum) sampled in Italy in 2009. The assay showed high sensitivity and reproducibility. Its application on bats screening resulted in a prevalence of 42%. This method could be suitable as screening tool in epidemiological surveys about the presence of bat-SARS-like CoVs, consequently to obtain a more realistic scenario of the viral prevalence in the population.


Reply to Rapid on‐site evaluation and the COVID‐19 pandemic


title: Clinical features of hemodialysis patients with COVID-19: a single-center retrospective study on 32 patients

text: OBJECTIVE: We retrospectively analyzed the data of 32 hemodialysis patients with COVID-19 to clarify the epidemiological characteristics of this special population. METHOD: The data of 32 hemodialysis patients with COVID-19, including epidemiological, demographic, clinical, laboratory, and radiological, were collected from the Blood Purification Department of Wuhan Fourth Hospital from February 3 to 16, 2020. RESULTS: Of the 32 patients, 23 were male, and the median age was 58 years; the median dialysis vintage was 33 months. Two groups were divided according to the patient’s primary renal disease: group 1 (16 patients with diabetic nephropathy), group 2 (12 patients with primary glomerulonephritis, 2 with obstructive kidney disease, 1 with hypertensive renal damage, and 1 with polycystic kidney). No significant differences were observed between the two groups in epidemiological characteristics, blood cell counts, and radiological performance. Hemodialysis patients are susceptible to COVID-19 at all ages, and patients with diabetes may be a high-risk population (50%). Common symptoms included fever (15 cases), cough (21 cases), and fatigue (7 cases). The blood lymphocyte count decreased in 84.6% of the patients (median: 0.765 × 10(9)/L). Chest CT revealed ground-glass-like lesions in 18 cases, unilateral lung patchiness in 7 cases, bilateral lung patchiness in 7 cases, and pleural effusion in 2 cases. CONCLUSION: Only 46.875% of the hemodialysis patients with COVID-19 had fever in the early stage; and diabetics may be the most susceptible population. A decrease in blood lymphocyte count and ground-glass opacity on chest CT scan is beneficial in identifying the high-risk population.


title: Sovereign Dignity, Nationalism and the Health of a Nation: A Study of China's Response in Combat of Epidemics

text: This paper seeks to understand the role of nationalism in China's policy towards the combat of emerging infectious diseases. By locating nationalism as a factor which facilitates or impedes global governance and international collaboration, this paper explores how nationalism influences China's political decision‐making. Given her historical experience, China has in its national psyche an impulse never to become ‘the sick man of the East’ again. Today, China's willingness to co‐operate with international bodies emanates out of reputational concerns rather than technical‐medical considerations. This was clearly manifested in her handling of two epidemics in recent years: the Severe Acute Respiratory Syndrome (SARS) and HIV/AIDS episodes. This paper concludes that China's nationalism plays an inhibiting role in China's attempts to further incorporate herself into the architecture of global health governance in the long run.


title: Antiviral activity of iridoid glycosides extracted from Fructus Gardeniae against influenza A virus by PACT-dependent suppression of viral RNA replication

text: Epidemic and pandemic influenza A virus (IAV) poses a significant threat to human populations worldwide. Iridoid glycosides are principal bioactive components from the Gardenia jasminoides J. Ellis fruit that exhibit antiviral activity against several strains of IAV. In the present study, we evaluated the protective effect of Fructus Gardeniae iridoid glycoside extracts (IGEs) against IAV by cytopathogenic effect(CPE), MTT and a plaque formation assay in vitro and examined the reduction in the pulmonary index (PI), restoration of body weight, reduction in mortality and increases in survival time in vivo. As a host factor, PACT provides protection against the pathogenic influenza A virus by interacting with IAV polymerase and activating the IFN-I response. To verify the whether IGEs suppress IAV replication in a PACT-dependent manner, IAV RNA replication, expression of PACT and the phosphorylation of eIF2α in A549 cells were detected; the levels of IFNβ, PACT and PKR in mouse lung tissues were determined; and the activity of IAV polymerase was evaluated in PACT-compromised cells. The results indicated that IGEs sufficiently alleviated cell damage and suppressed IAV replication in vitro, protecting mice from IAV-induced injury and lethal IAV infection. These anti-IAV effects might be related to disrupted interplay between IVA polymerase and PACT and/or prevention of a PACT-dependent overactivated IFN-I antiviral response. Taken together, our findings reveal a new facet of the mechanisms by which IGEs fight the influenza A virus in a PACT-dependent manner.


title: Peptides derived from HIV-1, HIV-2, Ebola virus, SARS coronavirus and coronavirus 229E exhibit high affinity binding to the formyl peptide receptor

text: Abstract Peptides derived from the membrane proximal region of fusion proteins of human immunodeficiency viruses 1 and 2, Coronavirus 229 E, severe acute respiratory syndrome coronavirus and Ebola virus were all potent antagonists of the formyl peptide receptor expressed in Chinese hamster ovary cells. Binding of viral peptides was affected by the naturally occurring polymorphisms at residues 190 and 192, which are located at second extracellular loop–transmembrane helix 5 interface. Substitution of R190 with W190 enhanced the affinity for a severe acute respiratory syndrome coronavirus peptide 6 fold but reduced the affinity for N-formyl-Nle–Leu–Phe by 2.5 fold. A 12 mer peptide derived from coronavirus 229E (ETYIKPWWVWL) was the most potent antagonist of the formyl peptide receptor W190 with a K i of 230 nM. Fluorescently labeled ETYIKPWWVWL was effectively internalized by all three variants with EC50 of ∼25 nM. An HKU-1 coronavirus peptide, MYVKWPWYVWL, was a potent antagonist but N-formyl-MYVKWPWYVWL was a potent agonist. ETYIKPWWVWL did not stimulate GTPγS binding but inhibited the stimulation by formyl-NleLeuPhe. It also blocked β arrestin translocation and receptor downregulation induced by formyl-Nle–Leu–Phe. This indicates that formyl peptide receptor may be important in viral infections and that variations in its sequence among individuals may affect their likelihood of viral and bacterial infections.


title: Cryptosporidiosis in buffalo calves (Bubalus bubalis): Prevalence and potential risk factors

text: The objective of the present study was to describe the prevalence and risk factors associated with cryptosporidiosis in buffalo calves in Middle Egypt. During one year, 458 fecal samples were collected from buffalo calves less than 3 month age in 55 small scale herds and examined for the presence of Cryptosporidium oocysts. Data describing age, gender, season, and herd management practices were gathered to assess potential risk factors. Fecal examination showed that 14.19% of the examined calves were positive for Cryptosporidium spp. Calves at 1–15 days were at the highest risk (P < 0.001), and a significant relationship between season and infection (P < 0.05) was recorded. A significant association between infection and hygiene (P < 0.001), type of floor (P < 0.01) and source of water (P < 0.01) was also recorded. Statistical analysis concerning the clinical signs and fecal characteristics revealed a significant association with fecal consistency (P < 0.001), presence of blood (P < 0.01) and mucous (P < 0.01). Moreover, a significant association was found between infection and the desire for suckling (P < 0.05) and tenesmus (P < 0.05). The results of the present study demonstrated the strong relation between infections by Cryptosporidium spp. and diarrhea in buffalo calves.


title: Safety considerations of chloroquine and hydroxychloroquine in treatment of COVID-19

text: Chloroquine and hydroxychloroquine are both used to treat COVID-19. Safety data in this specific population is largely unknown. In particular, cardiologic, gastro-intestinal and neuropsychiatric side-effects of (hydroxychloroquine) needs special attention in COVID-19 patients.


title: Evaluation of quantitative and type-specific real-time RT-PCR assays for detection of respiratory syncytial virus in respiratory specimens from children

text: Abstract Background: Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract morbidity in young children and immunosuppressed patients. Objectives: To rapidly and accurately quantify and subtype RSV in respiratory samples, we developed and evaluated two real-time RT-PCR assays. Study design: A quantitative assay was designed using primers for a consensus region of the matrix protein gene and a subtype-specific assay for RSV-A and RSV-B detection was designed using primers for the polymerase gene. Quantitative RSV RT-PCR results of pediatric nasal wash samples submitted to the University of Washington Virology Laboratory from December 2002, through May 2003, were compared to those of an indirect fluorescent antibody RSV antigen detection assay (FA). Results: Specificity of the RT-PCR assay was high, with no amplification of eleven common respiratory viruses and eight herpes viruses. Among 751 samples, RSV was detected in 267 (35.6%) by FA and in 286 (38.1%) by RT-PCR. Median RSV copy number in nasal wash samples that were positive by both FA and RT-PCR was 2.5×107 copies/mL versus a median of 3.0×104 copies/mL for samples positive by RT-PCR only (P<0.001). The detection and quantity of RSV in respiratory specimens was associated with younger age, but not with gender or hospitalization. Among positive samples from this Seattle cohort, 52% were subtype A and 48% were subtype B. Both subtypes were detected with similar viral loads among all patient groups (stratified by age, gender, and hospitalization), and throughout the specimen collection period. Conclusions: These real-time RT-PCR assays provide a rapid, specific, and highly sensitive alternative for detecting, quantifying, and subtyping RSV in clinical specimens.


title: Differentiation of Bovine Coronavirus (Bcov) Genotypes by a Restriction Enzyme Assay

text: This article reports the use of the GsuI restriction enzyme to differentiate genotypes of Bovine Coronavirus (BCoV), based on an 18-nucleotide deletion of S1-coding region found in one of the two genotypes. It was concluded that this assay can be used as a rapid tool for BCoV genotypes differentiation.


title: Computed tomography findings in a case of coronavirus disease 2019

text: Abstract Coronavirus disease 2019 (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is mainly characterized by pulmonary inflammation. The preferred imaging modality is chest computed tomography (CT) which plays an important role in early diagnosis, quarantine, and treatment as well as in the evaluation of therapeutic efficacy. We report the imaging data from a confirmed case of COVID-19 admitted to our hospital. Our aims are to improve understanding of this disease and to facilitate early diagnosis and evaluation of therapeutic efficacy. A 70-year-old woman living in the epidemic area presented with a 2-day history of intermittent fever. Chest CT revealed multiple ground glass opacities in both lungs, mainly distributed subpleurally and in the middle and lateral lung fields, particularly in the right lung, where they had partially fused into patches. Local interlobular septal thickening was also observed. Throat swabs were positive for the SARS-CoV-2 nucleic acid, which confirmed the diagnosis of COVID-19. Chest CT plays a key role in the diagnosis of COVID-19, providing an accurate diagnosis and is a sensitive technique for evaluation of therapeutic efficacy. It has the advantages of promptness, convenience, and high efficiency.


title: Genetic analysis of heptad-repeat regions in the G2 fusion subunit of the Junín arenavirus envelope glycoprotein

text: Abstract The G2 fusion subunit of the Junín virus envelope glycoprotein GP-C contains two hydrophobic heptad-repeat regions that are postulated to form a six-helix bundle structure required for the membrane fusion activity of Class I viral fusion proteins. We have investigated the role of these heptad-repeat regions and, specifically, the importance of the putative interhelical a and d position sidechains by using alanine-scanning mutagenesis. All the mutant glycoproteins were expressed and transported to the cell surface. Proteolytic maturation at the subtilisin kexin isozyme-1/site-1-protease (SKI-1/S1P) cleavage site was observed in all but two of the mutants. Among the adequately cleaved mutant glycoproteins, four positions in the N-terminal region (I333, L336, L347 and L350) and two positions in the C-terminal region (R392 and W395) were shown to be important determinants of cell–cell fusion. Taken together, our results indicate that α-helical coiled-coil structures are likely critical in promoting arenavirus membrane fusion. These findings support the inclusion of the arenavirus GP-C among the Class I viral fusion proteins and suggest pharmacologic and immunologic strategies for targeting arenavirus infection and hemorrhagic fever.


title: Strategies for managing risk in a changing aviation environment

text: Abstract Given the increasing volatility in the economic performance of airlines, partially reflecting the dynamics of demand for air transport and the fixed costs associated with the industry, all stakeholders need to consider appropriate strategies for better managing the risks. Many risks were identified in the literature previously, some even decades ago, however most have yet to be satisfactorily addressed. Urgency is growing. Removal of the remaining barriers to competition at all levels, congestion management, open skies policies across continents, computer-centric air traffic management systems and increased research and development into the processes and technology needed to reduce environmental externalities remain among the top challenges for the next decade.


title: Transformation of cross-boundary governance in the Greater Pearl River Delta, China: Contested geopolitics and emerging conflicts

text: Drawing upon the perspective of state rescaling in city-region governance, this paper explores the transformation of cross-boundary governance in the Greater Pearl River Delta (GPRD), a cross-boundary region on the South China coast consisting of the PRD under the jurisdiction of Guangdong Province, and Hong Kong and Macao Special Administrative Regions (SARs). Based on updated field investigation and in-depth interviews between 2008 and 2012, the study argues that the Greater PRD has undergone dramatic restructuring of regional governance since mid 2000s, reflected by rescaling attempts initiated by Guangdong provincial government and bottom-up resistance from local residents in Hong Kong. The interaction of the rescaling dynamics has engendered the contested geopolitics of the cross-boundary governance as the regional integration intensified. Notably, the complexity of cross-boundary governance has been heightened by emerging conflicts, resulted from the increasing number of mainlanders as ‘individual visitors’ to Hong Kong since the implementation of the Closer Economic Partnership Arrangement (CEPA), the first bilateral free trade agreement between Hong Kong and China effective from 2004. By examining the Greater PRD as a salient mega city-region, the present study enriches the growing literature on state rescaling and cross-border governance in contemporary globalization.


title: Report from the COVID-19 Virtual Summit, Disaster Experts Speak Out, March 31, 2020

text: This article captures the webinar narrative on March 31, 2020 of four expert panelists addressing three questions on the current coronavirus disease 2019 (COVID-19) pandemic. Each panelist was selected for their unique personal expertise, ranging from front-line emergency physicians from multiple countries, an international media personality, former director of the US Strategic National Stockpile, and one of the foremost international experts in disaster medicine and public policy. The forum was moderated by one of the most widely recognized disaster medical experts in the world. The four panelists were asked three questions regarding the current pandemic as follows: 1. What do you see as a particular issue of concern during the current pandemic? 2. What do you see as a particular strength during the current pandemic? 3. If you could change one thing about the way that the pandemic response is occurring, what would you change?


title: JNK and p38 mitogen-activated protein kinase pathways contribute to porcine epidemic diarrhea virus infection

text: Abstract The mitogen-activated protein kinase (MAPK) pathways, which are central building blocks in the intracellular signaling network, are often manipulated by viruses of diverse families to favor their replication. Among the MAPK family, the extracellular signal-regulated kinase (ERK) pathway is known to be modulated during the infection with porcine epidemic diarrhea virus (PEDV); however, involvement of stress-activated protein kinases (SAPKs) comprising p38 MAPK and c-Jun NH2-terminal kinase (JNK) remains to be determined. Therefore, in the present study, we investigated whether activation of p38 MAPK and JNK cascades is required for PEDV replication. Our results showed that PEDV activates p38 MAPK and JNK1/2 up to 24h post-infection, whereas, thereafter their phosphorylation levels recede to baseline levels or even fall below them. Notably, UV-irradiated inactivated PEDV, which can enter cells but cannot replicate inside them, failed to induce phosphorylation of p38 MAPK and JNK1/2 suggesting that viral biosynthesis is essential for activation of these kinases. Treatment of cells with selective p38 or JNK inhibitors markedly impaired PEDV replication in a dose-dependent manner and these antiviral effects were found to be maximal during the early times of the infection. Furthermore, direct pharmacological inhibition of p38 MAPK or JNK1/2 activation resulted in a significant reduction of viral RNA synthesis, viral protein expression, and progeny release. However, independent treatments with either SAPK inhibitor did not inhibit PEDV-induced apoptotic cell death mediated by activation of mitochondrial apoptosis-inducing factor (AIF) suggesting that SAPKs are irrelevant to the apoptosis pathway during PEDV infection. In summary, our data demonstrated critical roles of the p38 and JNK1/2 signaling pathways in facilitating successful viral infection during the post-entry steps of the PEDV life cycle.


title: Acute limb ischemia in patients with COVID-19 pneumonia

text: OBJECTIVES: The aim of our study is to describe the incidence and characteristics, as well as clinical outcomes of patients presenting and treated for acute limb ischemia (ALI) in patients with novel coronavirus (COVID-19) infection during the 2020 coronavirus pandemic. METHODS: This is a single center, observational cohort study. Data of all patients tested positive with COVID-19 and presented with ALI vascular diseases requiring urgent operative treatment were collected in a prospectively maintained database. For this series, successful revascularization of the treated arterial segment was defined as the absence of early (<30 days) re-occlusion, or major amputation or death ≤24 hours. Primary outcomes were successful revascularization, early (≤30 days) and late survival (≥30 days), postoperative (≤ 30 days) complications, and limb salvage. RESULTS: We evaluated 20 patients with ALI who were positive for COVID-19. Considering the period January-to-March, the incidence rate of patients presenting with ALI in 2020 was significantly higher than 2019 [23/141 (16.3%) vs. 3/163 (1.8%), P < 0.001)]. There were 18 (90%) male and 2 female patients. Mean age was 75 years ± 9 (range, 62-95). All patients had already been diagnosed with COVID-19 pneumonia. Operative treatment was performed in 17 (85%) patients. Revascularization was successful in 12/17 (70.6%) patients. Although successful revascularization was not significantly associated with the postoperative use of intravenous heparin (64.7% vs. 83.3%, P = 0.622), no patients receiving intravenous heparin required reintervention. There were 8 (40%) in-hospital mortalities. Age was significantly higher in those who died (years, 81 ± 10 vs. 71 ± 5, P = 0.008). The use of continued postoperative systemic heparin infusion was significantly associated with survival (0% vs. 57.1%, P = 0.042). CONCLUSIONS: In our preliminary experience, the incidence of ALI has significantly increased during COVID-19 pandemic in the Italian Lombardia region. Successful revascularization was lower than expected which we hypothesize is due to a virus-related hypercoagulable state. The use of prolonged systemic heparin may improve surgical treatment efficacy as well as improve limb salvage and overall mortality.


title: Binding of the 5′-untranslated region of coronavirus RNA to zinc finger CCHC-type and RNA-binding motif 1 enhances viral replication and transcription

text: Coronaviruses RNA synthesis occurs in the cytoplasm and is regulated by host cell proteins. In a screen based on a yeast three-hybrid system using the 5′-untranslated region (5′-UTR) of SARS coronavirus (SARS-CoV) RNA as bait against a human cDNA library derived from HeLa cells, we found a positive candidate cellular protein, zinc finger CCHC-type and RNA-binding motif 1 (MADP1), to be able to interact with this region of the SARS-CoV genome. This interaction was subsequently confirmed in coronavirus infectious bronchitis virus (IBV). The specificity of the interaction between MADP1 and the 5′-UTR of IBV was investigated and confirmed by using an RNA pull-down assay. The RNA-binding domain was mapped to the N-terminal region of MADP1 and the protein binding sequence to stem–loop I of IBV 5′-UTR. MADP1 was found to be translocated to the cytoplasm and partially co-localized with the viral replicase/transcriptase complexes (RTCs) in IBV-infected cells, deviating from its usual nuclear localization in a normal cell using indirect immunofluorescence. Using small interfering RNA (siRNA) against MADP1, defective viral RNA synthesis was observed in the knockdown cells, therefore indicating the importance of the protein in coronaviral RNA synthesis.


title: Is BMI higher in younger patients with COVID‐19? Association between BMI and COVID‐19 hospitalization by age

text: OBJECTIVE: Obesity has been found to be a risk factor for hospitalization with COVID‐19. We were interested in understanding whether patients hospitalized with COVID‐19 differed in BMI at older versus younger ages, and if trends were independent of diabetes and hypertension. METHODS: We performed a cross‐sectional analysis of patients hospitalized with moderate to severe COVID‐19 at Northwestern Memorial Hospital from March 19(th), 2020 until April 4(th), 2020. We compared patients hospitalized with COVID‐19 above and below the age of 50, and to those hospitalized without COVID‐19. RESULTS: We found patients younger than 50 years of age hospitalized with COVID‐19 without diabetes or hypertension had mean BMI greater than those older than 50 years of age, with BMI 43.1 (95%CI 34.5 – 51.7) kg/m(2) vs 30.1 (95%CI 27.7 – 32.5) kg/m(2) (p=0.02). Furthermore, BMI appears to inversely correlate with increasing age amongst patients hospitalized with COVID‐19. We did not detect the same difference or trend for patients hospitalized without COVID‐19. CONCLUSION: We found younger patients (age <50 years) with COVID‐19 had higher mean BMI than older patients with COVID‐19, with and without diabetes and hypertension. This trend did not exist in patients without COVID‐19 hospitalized during the same time‐period.


title: Does Viral Co-Infection Influence the Severity of Acute Respiratory Infection in Children?

text: BACKGROUND: Multiple viruses are often detected in children with respiratory infection but the significance of co-infection in pathogenesis, severity and outcome is unclear. OBJECTIVES: To correlate the presence of viral co-infection with clinical phenotype in children admitted with acute respiratory infections (ARI). METHODS: We collected detailed clinical information on severity for children admitted with ARI as part of a Spanish prospective multicenter study (GENDRES network) between 2011–2013. A nested polymerase chain reaction (PCR) approach was used to detect respiratory viruses in respiratory secretions. Findings were compared to an independent cohort collected in the UK. RESULTS: 204 children were recruited in the main cohort and 97 in the replication cohort. The number of detected viruses did not correlate with any markers of severity. However, bacterial superinfection was associated with increased severity (OR: 4.356; P-value = 0.005), PICU admission (OR: 3.342; P-value = 0.006), higher clinical score (1.988; P-value = 0.002) respiratory support requirement (OR: 7.484; P-value < 0.001) and longer hospital length of stay (OR: 1.468; P-value < 0.001). In addition, pneumococcal vaccination was found to be a protective factor in terms of degree of respiratory distress (OR: 2.917; P-value = 0.035), PICU admission (OR: 0.301; P-value = 0.011), lower clinical score (-1.499; P-value = 0.021) respiratory support requirement (OR: 0.324; P-value = 0.016) and oxygen necessity (OR: 0.328; P-value = 0.001). All these findings were replicated in the UK cohort. CONCLUSION: The presence of more than one virus in hospitalized children with ARI is very frequent but it does not seem to have a major clinical impact in terms of severity. However bacterial superinfection increases the severity of the disease course. On the contrary, pneumococcal vaccination plays a protective role.


title: Community-Wide, Contemporaneous Circulation of a Broad Spectrum of Human Rhinoviruses in Healthy Australian Preschool-Aged Children During a 12-Month Period

text: Human rhinovirus (HRV) replication triggers exacerbation of asthma and causes most acute respiratory illnesses (ARIs), which may manifest as influenza-like illness. The recent assignment of 60 previously unknown HRV types to a third HRV species, Human rhinovirus C, raised questions about the prevalence of these picornavirus types in the community, the extent of HRV diversity at a single site, and whether the HRVs have an equally diverse clinical impact on their hosts. We quantified HRV diversity, and there was no clinical impact attributable to HRV species and genotypes among a community population of preschool-aged children with ARI who provided respiratory samples during 2003. All HRV species were represented among 138 children with ARI, and 74 distinct HRV types were cocirculating. Fever accompanied 32.8% of HRV-positive ARI cases. HRVs were less likely than DNA viruses to be codetected with another virus, suggesting virus interference at the community level, demonstrated by the inverse correlation between influenza virus detection and HRV detection.


La chirurgie gynécologique à l’épreuve de l’infection à COVID-19. Quelles recommandations formuler?


title: Enhanced inflammation in New Zealand white rabbits when MERS-CoV reinfection occurs in the absence of neutralizing antibody

text: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic betacoronavirus that was first detected in humans in 2012 as a cause of severe acute respiratory disease. As of July 28, 2017, there have been 2,040 confirmed cases with 712 reported deaths. While many infections have been fatal, there have also been a large number of mild or asymptomatic cases discovered through monitoring and contact tracing. New Zealand white rabbits are a possible model for asymptomatic infection with MERS-CoV. In order to discover more about non-lethal infections and to learn whether a single infection with MERS-CoV would protect against reinfection, we inoculated rabbits with MERS-CoV and monitored the antibody and inflammatory response. Following intranasal infection, rabbits developed a transient dose-dependent pulmonary infection with moderately high levels of viral RNA, viral antigen, and perivascular inflammation in multiple lung lobes that was not associated with clinical signs. The rabbits developed antibodies against viral proteins that lacked neutralizing activity and the animals were not protected from reinfection. In fact, reinfection resulted in enhanced pulmonary inflammation, without an associated increase in viral RNA titers. Interestingly, passive transfer of serum from previously infected rabbits to naïve rabbits was associated with enhanced inflammation upon infection. We further found this inflammation was accompanied by increased recruitment of complement proteins compared to primary infection. However, reinfection elicited neutralizing antibodies that protected rabbits from subsequent viral challenge. Our data from the rabbit model suggests that people exposed to MERS-CoV who fail to develop a neutralizing antibody response, or persons whose neutralizing antibody titers have waned, may be at risk for severe lung disease on re-exposure to MERS-CoV.


title: Therapeutic Applications of Rose Hips from Different Rosa Species

text: Rosa species, rose hips, are widespread wild plants that have been traditionally used as medicinal compounds for the treatment of a wide variety of diseases. The therapeutic potential of these plants is based on its antioxidant effects caused by or associated with its phytochemical composition, which includes ascorbic acid, phenolic compounds and healthy fatty acids among others. Over the last few years, medicinal interest in rose hips has increased as a consequence of recent research that has studied its potential application as a treatment for several diseases including skin disorders, hepatotoxicity, renal disturbances, diarrhoea, inflammatory disorders, arthritis, diabetes, hyperlipidaemia, obesity and cancer. In this review, the role of different species of Rosa in the prevention of treatment of various disorders related to oxidative stress, is examined, focusing on new therapeutic approaches from a molecular point of view.


title: Vomiting and wasting disease associated with hemagglutinating encephalomyelitis viruses infection in piglets in jilin, china

text: One coronavirus strain was isolated from brain tissues of ten piglets with evident clinical manifestations of vomiting, diarrhea and dyskinesia in Jilin province in China. Antigenic and genomic characterizations of the virus (isolate PHEV-JLsp09) were based on multiplex PCR and negative staining electron microscopy and sequence analysis of the Hemagglutinin-esterase (HE) gene. These piglets were diagnosed with Porcine hemagglutinating encephalomyelitis virus (PHEV). Necropsy was performed on the piglets. Major pathological changes included meningeal hyperemia, meningeal hemorrhage and cortical hemorrhage. Minor changes were also observed in other organs. Histopathological changes included satellitosis and neuronophagia in the cerebral cortex. Mice were infected with the isolated virus. Their histopathological changes were similar to those symptoms observed in the piglets, exhibiting typical changes for non-suppurative encephalitis. Thus, Porcine hemagglutinating encephalomyelitis virus mainly causes damage to the nervous system but also impacts other organs. This viral strain (isolate PHEV-JLsp09) found in the Siping area of Jilin Province in China is evolutionally closest to the HEV-67N stain (North American strain), indicating that this viral strain evolved from the PHEV from North America.


title: A New Bug on the Block

text: In December 2019, clusters of pneumonia cases started showing up in the Hubei region of China, especially in its largest city Wuhan. As these piled up, researchers at the Chinese Academy of Sciences’ laboratories and the Chinese Centre for Disease Control started looking for the cause. This turned out to be a new virus, whose genetic sequence matched very closely with two coronaviruses isolated from bats in China in 2018.


title: Rapid peptide-based screening on the substrate specificity of severe acute respiratory syndrome (SARS) coronavirus 3C-like protease by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) 3C-like protease (3CL(pro)) mediates extensive proteolytic processing of replicase polyproteins, and is considered a promising target for anti-SARS drug development. Here we present a rapid and high-throughput screening method to study the substrate specificity of SARS-CoV 3CL(pro). Six target amino acid positions flanking the SARS-CoV 3CL(pro) cleavage site were investigated. Each batch of mixed peptide substrates with defined amino acid substitutions at the target amino acid position was synthesized via the “cartridge replacement” approach and was subjected to enzymatic cleavage by recombinant SARS-CoV 3CL(pro). Susceptibility of each peptide substrate to SARS-CoV 3CL(pro) cleavage was monitored simultaneously by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The hydrophobic pocket in the P2 position at the protease cleavage site is crucial to SARS-CoV 3CL(pro)-specific binding, which is limited to substitution by hydrophobic residue. The binding interface of SARS-CoV 3CL(pro) that is facing the P1′ position is suggested to be occupied by acidic amino acids, thus the P1′ position is intolerant to acidic residue substitution, owing to electrostatic repulsion. Steric hindrance caused by some bulky or β-branching amino acids in P3 and P2′ positions may also hinder the binding of SARS-CoV 3CL(pro). This study generates a comprehensive overview of SARS-CoV 3CL(pro) substrate specificity, which serves as the design basis of synthetic peptide-based SARS-CoV 3CL(pro) inhibitors. Our experimental approach is believed to be widely applicable for investigating the substrate specificity of other proteases in a rapid and high-throughput manner that is compatible for future automated analysis.


title: Towards a conceptual framework to support one-health research for policy on emerging zoonoses

text: Summary In the past two decades there has been a growing realisation that the livestock sector was in a process of change, resulting from an expansion of intensive animal production systems and trade to meet a globalised world's increasing demand for livestock products. One unintended consequence has been the emergence and spread of transboundary animal diseases and, more specifically, the resurgence and emergence of zoonotic diseases. Concurrent with changes in the livestock sector, contact with wildlife has increased. This development has increased the risk of transmission of infections from wildlife to human beings and livestock. Two overarching questions arise with respect to the real and perceived threat from emerging infectious diseases: why are these problems arising with increasing frequency, and how should we manage and control them? A clear conceptual research framework can provide a guide to ensure a research strategy that coherently links to the overarching goals of policy makers. We propose such a new framework in support of a research and policy-generation strategy to help to address the challenges posed by emerging zoonoses.


title: A Serological Survey for Avian Infectious Bronchitis Virus and Newcastle Disease Virus Antibodies in Backyard (Free-range) Village Chickens in Mexico

text: The commercial flocks in Yucatan, Mexico are free of Newcastle disease virus (NDV) in its velogenic viscerotropic form, but little is known about the disease status of backyard poultry. A seroprevalence survey in 30 villages using haemagglutination inhibition (HI) tests for infectious bronchitis virus (IBV) and NDV antibodies was carried out from December 1997 to June 1998. The seroprevalences were 56.5% (95% CI 50–63%) for IBV and 2.2% (95% CI 0.5–3.8%) for NDV. All the villages had chickens that were positive for antibodies to IBV and nine of the villages had chickens that were positive for antibodies to NDV. This suggests that IBV may be responsible for a large proportion of the respiratory disease observed in backyard chickens in Yucatan. The implications of these findings are discussed, including the highly susceptible status of the backyard chickens in Yucatan to NDV and the possibility of this virus being one cause of the syndrome known as mortandad by the local people.


COVID‐19, Boko Haram and the Pursuit of Survival: A Battle of Lives against Livelihoods


title: Strategies to develop antivirals against enterovirus 71

text: Enterovirus 71 (EV71) is an important human pathogen which may cause severe neurological complications and death in children. The virus caused several outbreaks in the Asia-Pacific region during the past two decades and has been considered a significant public health problem in the post-poliovirus eradication era. Unlike poliovirus, there is no effective vaccine or approved antivirals against EV71. To explore anti-EV71 agents therefore is of vital importance. Several strategies have been employed to develop antivirals based on the molecular characteristics of the virus. Among these, some small molecules that were developed against human rhinoviruses and poliovirus are under evaluation. In this review, we discuss the recent development of such small molecules against EV71, known drug resistance and possible solutions to it, and animal models for evaluating the efficacy of these antivirals. Although further investigation is required for clinical applications of the existing candidates, the molecular mechanisms revealed for the inhibition of EV71 replication can be used for designing new molecules against this virus in the future.


title: Xenosurveillance reflects traditional sampling techniques for the identification of human pathogens: A comparative study in West Africa

text: BACKGROUND: Novel surveillance strategies are needed to detect the rapid and continuous emergence of infectious disease agents. Ideally, new sampling strategies should be simple to implement, technologically uncomplicated, and applicable to areas where emergence events are known to occur. To this end, xenosurveillance is a technique that makes use of blood collected by hematophagous arthropods to monitor and identify vertebrate pathogens. Mosquitoes are largely ubiquitous animals that often exist in sizable populations. As well, many domestic or peridomestic species of mosquitoes will preferentially take blood-meals from humans, making them a unique and largely untapped reservoir to collect human blood. METHODOLOGY/PRINCIPAL FINDINGS: We sought to take advantage of this phenomenon by systematically collecting blood-fed mosquitoes during a field trail in Northern Liberia to determine whether pathogen sequences from blood engorged mosquitoes accurately mirror those obtained directly from humans. Specifically, blood was collected from humans via finger-stick and by aspirating bloodfed mosquitoes from the inside of houses. Shotgun metagenomic sequencing of RNA and DNA derived from these specimens was performed to detect pathogen sequences. Samples obtained from xenosurveillance and from finger-stick blood collection produced a similar number and quality of reads aligning to two human viruses, GB virus C and hepatitis B virus. CONCLUSIONS/SIGNIFICANCE: This study represents the first systematic comparison between xenosurveillance and more traditional sampling methodologies, while also demonstrating the viability of xenosurveillance as a tool to sample human blood for circulating pathogens.


Full spectrum of COVID-19 severity still being depicted – Authors' reply


title: Gene expression patterns in peripheral blood leukocytes in patients with recurrent ciguatera fish poisoning: Preliminary studies

text: Abstract Ciguatera fish poisoning (ciguatera) is a common clinical syndrome in areas where there is dependence on tropical reef fish for food. A subset of patients develops recurrent and, in some instances, chronic symptoms, which may result in substantial disability. To identify possible biomarkers for recurrent/chronic disease, and to explore correlations with immune gene expression, peripheral blood leukocyte gene expression in 10 ciguatera patients (7 recurrent and 3 acute) from the U.S. Virgin Islands, and 5 unexposed Florida controls were evaluated. Significant differences in gene expression were noted when comparing ciguatera patients and controls; however, it was not possible to differentiate between patients with acute and recurrent disease, possibly due to the small sample sizes involved.


title: Discovery of a subgenotype of human coronavirus NL63 associated with severe lower respiratory tract infection in China, 2018

text: Human coronavirus NL63 (HCoV-NL63) is primarily associated with common cold in children, elderly and immunocompromised individuals. Outbreaks caused by HCoV-NL63 are rare. Here we report a cluster of HCoV-NL63 cases with severe lower respiratory tract infection that arose in Guangzhou, China, in 2018. Twenty-three hospitalized children were confirmed to be HCoV-NL63 positive, and most of whom were hospitalized with severe pneumonia or acute bronchitis. Whole genomes of HCoV-NL63 were obtained using next-generation sequencing. Phylogenetic and single amino acid polymorphism analyses showed that this outbreak was associated with two subgenotypes (C3 and B) of HCoV-NL63. Half of patients were identified to be related to a new subgenotype C3. One unique amino acid mutation at I507 L in spike protein receptor binding domain (RBD) was detected, which segregated this subgenotype C3 from other known subgenotypes. Pseudotyped virus bearing the I507 L mutation in RBD showed enhanced entry into host cells as compared to the prototype virus. This study proved that HCoV-NL63 was undergoing continuous mutation and has the potential to cause severe lower respiratory disease in humans.


title: Roosting ecology and variation in adaptive and innate immune system function in the Brazilian free-tailed bat (Tadarida brasiliensis)

text: Bats have recently been implicated as reservoirs of important emerging diseases. However, few studies have examined immune responses in bats, and even fewer have evaluated these responses in an ecological context. We examined aspects of both innate and adaptive immune response in adult female Brazilian free-tailed bats (Tadarida brasiliensis) at four maternity roosts (two natural caves and two human-made bridges) in south-central Texas. Immune measurements included in vitro bactericidal ability of whole blood and in vivo T cell mediated response to mitogenic challenge. Bactericidal activity in T. brasiliensis varied with roosting ecology, but appears to be sensitive to colony-level effects. Blood from females living at one cave had significantly lower bactericidal ability than blood from females at three other sites. T cell mediated response in this species was associated with variation in roost ecology, with females from two caves having greater responses than females from two bridges. T cell mediated response and bactericidal activity were negatively correlated with one another within individuals that were tested for both. Variation in immunological response of T. brasiliensis is important for understanding the influence of the environment on the frequency and distribution of immunologically competent individuals and for understanding disease-host dynamics in this and other colonial species.


title: Effects of information-dependent vaccination behavior on coronavirus outbreak: insights from a SIRI model

text: A mathematical model is proposed to assess the effects of a vaccine on the time evolution of a coronavirus outbreak. The model has the basic structure of SIRI compartments (susceptible–infectious–recovered–infectious) and is implemented by taking into account of the behavioral changes of individuals in response to the available information on the status of the disease in the community. We found that the cumulative incidence may be significantly reduced when the information coverage is high enough and/or the information delay is short, especially when the reinfection rate is high enough to sustain the presence of the disease in the community. This analysis is inspired by the ongoing outbreak of a respiratory illness caused by the novel coronavirus COVID-19.


title: Considerations for interactions of drugs used for the treatment of COVID-19 with anti-Cancer treatments

text: SARS-CoV2 infection is an emerging issue worldwide. Cancer patient are at increased risk of infection compared to general population. On the other hand, these patients are at major risk of drug interactions caused by renal and hepatic impairment background. Because of the long-term use of chemotherapy drugs, drug interactions are important in these patients especially with SARS-CoV2 treatments now. This paper is review of reported drug interactions of current treatments for COVID-19 and anticancer agents.


title: Korean MERS: A new cross continent emerging infectious disease

text: Abstract Middle East respiratory syndrome is a new emerging infectious disease that was firstly detected in the Middle East. This disease can cause severe respiratory illness and was proved to be a coronavirus infection. The infection was firstly confined within the Middle East. However, the new emergence of this infection is in Korea and Korean Middle East respiratory syndrome becomes the new concern in public health. In this brief article, the authors discuss on this new cross continent emerging infectious disease.


title: Novel Respiratory Virus Infections in Children, Brazil

text: Recently discovered respiratory viruses were detected in 19 (9.2%) of 205 nasal swab specimens from children in Brazil with respiratory illnesses. Five each were positive for human metapneumovirus (HMPV) alone and human bocavirus (HBoV) alone, 3 for human coronaviruses (HCoV-HKU1 or -NL63) alone, and 6 for more than 1 recently discovered virus.


title: Evidence for zoonotic origins of Middle East respiratory syndrome coronavirus

text: Middle East respiratory syndrome (MERS) is an emerging infectious disease, caused by Middle East respiratory syndrome coronavirus (MERS-CoV) and is considered to be a zoonosis. However, the natural reservoirs of MERS-CoV remain obscure, with bats and camels as the most suspected sources. In this article, we review the evidence supporting a bat/camel origin of human MERS-CoV infection and current knowledge on the modes of camel-to-human transmission of MERS-CoV.


title: Chapter 4 Type I Interferon Modulates the Battle of Host Immune System Against Viruses

text: Abstract Type I interferon (IFN), as its name implies, ‘interferes’ with virus replication by activating numerous genes. Further, virus-induced type I IFN regulates the magnitude and functions of cells directing the host immune system. Importantly, recent exploration into how type I IFN operates following virus infection has advanced our understanding of its role with respect to modulation of host innate and adaptive immune responses. Such activities include the activation of antigen-presenting dendritic cells and the localization, expansion or differentiation of virus-specific T lymphocytes and antibody-producing B lymphocytes. However, type I IFN not only benefits the host but can also induce unnecessary or extremely pathogenic immune responses. This review focuses on such interactions and the manner in which type I IFN induces dynamic changes in the host immune network, particularly adaptive immune responses to viral invasion. Manipulating the type I IFN-mediated host immune response during virus infections could provide new immunotherapeutic interventions to remedy viral diseases and implement more effective and sustainable type I IFN therapy.


title: Development and validation of an LC-MS/MS method for detection and quantification of in vivo derived metabolites of [Pyr(1)]apelin-13 in humans

text: [Pyr(1)]apelin-13 is the predominant apelin peptide isoform in the human cardiovascular system and plasma. To date, few studies have investigated [Pyr(1)]apelin-13 metabolism in vivo in rats with no studies examining its stability in humans. We therefore aimed to develop an LC-MS/MS method for detection and quantification of intact [Pyr(1)]apelin-13 and have used this method to identify the metabolites generated in vivo in humans. [Pyr(1)]apelin-13 (135 nmol/min) was infused into six healthy human volunteers for 120 minutes and blood collected at time 0 and 120 minutes after infusion. Plasma was extracted in the presence of guanidine hydrochloride and analysed by LC-MS/MS. Here we report a highly sensitive, robust and reproducible method for quantification of intact [Pyr(1)]apelin-13 and its metabolites in human plasma. Using this method, we showed that the circulating concentration of intact peptide was 58.3 ± 10.5 ng/ml after 120 minutes infusion. We demonstrated for the first time that in humans, [Pyr(1)]apelin-13 was cleaved from both termini but the C-terminal was more susceptible to cleavage. Consequently, of the metabolites identified, [Pyr(1)]apelin-13((1–12)), [Pyr(1)]apelin-13((1–10)) and [Pyr(1)]apelin-13((1–6)) were the most abundant. These data suggest that apelin peptides designed for use as cardiovascular therapeutics, should include modifications that minimise C-terminal cleavage.


title: Dianke virus: A new mesonivirus species isolated from mosquitoes in Eastern Senegal

text: Abstract An increasing number of insect-specific viruses are found around the world. Very recently, a new group of insect-specific viruses, the Mesoniviridae family, was discovered in Africa, Asia, North America and Australia. Here we report the first detection and isolation of a new virus belonging to Mesonivirus genus in Senegal, West Africa. The so-called Dianke virus was detected in 21 species of arthropods trapped in the eastern part of the country. Male individuals were also infected, supporting vertical transmission assertion of insect specific viruses. As described for other mesoniviruses, no viral replication was observed after inoculation of mammalian cells. Viral replication in mosquito cells was blocked at a temperature of 37 °C, highlighting the importance of thermal conditions in Mesonivirus host restriction. Similar to our study, where a diverse range of arthropod vectors were found infected by the new virus, several studies have detected mesonivirus infection in mosquitoes with concerns for human health. It has been shown that dual infections in mosquito can alter viral infectivity. Due to their extensive geographic distribution and host range, as well as their use as potential disease control agents in vector populations, more studies should be done for a better knowledge of arthropod-restricted viruses prevalence and diversity.


title: Increased viral titers and enhanced reactivity of antibodies to the spike glycoprotein of murine coronavirus produced by infection at pH 6

text: Abstract Infection of cell monolayers by murine coronavirus A59 at pH 6 rather than 7 yielded a ten-fold increase in the infectious titer and a remarkable enhancement of the reactivities of monoclonal and polyclonal antibodies against the spike glycoprotein in immunoblotting, immuno-precipitation and enzyme-linked immunosorbent assays. These observations are very useful for detecting antibodies against the S glycoprotein of coronaviruses and enhancing infectious titers.


Getting Ahead of the Curve: How Ochsner Became a Leader in SARS-CoV-2 Diagnostic Testing


title: Fever screening of seasonal influenza patients using a cost-effective thermopile array with small pixels for close-range thermometry

text: Summary Objective Infrared thermography systems have been used for fever screening at many airports since the outbreak of severe acute respiratory syndrome (SARS) in 2003. However, many of these systems are expensive and non-portable. Therefore, we developed a cost-effective and compact (2.9 × 5.8 × 2.0cm) thermopile array for fever screening of patients with infectious diseases in the clinical setting. Methods The array was created with small pixels (48 × 47 = 2256 pixels) fabricated on a silicon wafer using microelectromechanical systems technology. We tested this array on 155 febrile and afebrile patients (35.4°C ≤ axillary temperature ≤ 39.3°C) with seasonal influenza at the Japan Self-Defense Forces Central Hospital. Results The maximum facial temperature, measured by the array at 0.3 m from the subject, exhibited a positive correlation with axillary temperature measured using a contact-type thermometer (r = 0.71, p < 0.01). The sensitivity and specificity of the thermopile array in identifying the febrile subjects were 80.5% and 93.3%, respectively, setting the threshold cut-off of maximum facial temperature at an appropriate value. Conclusions Our cost-effective thermopile array appears promising for future close-range fever screening of patients with infectious diseases at primary care doctor clinics, health care centers, and quarantine stations in developing and developed countries.


title: Metabolic Engineering of Glycyrrhizin Pathway by Over-Expression of Beta-amyrin 11-Oxidase in Transgenic Roots of Glycyrrhiza glabra

text: Glycyrrhiza glabra is one of the most important and well-known medicinal plants which produces various triterpene saponins such as glycyrrhizin. Beta-amyrin 11-oxidase (CYP88D6) plays a key role in engineering pathway of glycyrrhizin production and converts an intermediated beta-amyrin compound to glycyrrhizin. In this study, pBI121(GUS-9):CYP88D6 construct was transferred to G. glabra using Agrobacterium rhizogene ATCC 15834. The quantitation of transgene was measured in putative transgenic hairy roots using qRT-PCR. The amount of glycyrrhizin production was measured by HPLC in transgenic hairy root lines. Gene expression analysis demonstrated that CYP88D6 was over-expressed only in one of transgenic hairy root lines and was reduced in two others. Beta-amyrin 24-hydroxylase (CYP93E6) was significantly expressed in one of the control hairy root lines. The amount of glycyrrhizin metabolite in over-expressed line was more than or similar to that of control hairy root lines. According to the obtained results, it would be recommended that multi-genes of glycyrrhizin biosynthetic pathway be transferred simultaneously to the hairy root in order to increase glycyrrhizin content.


title: High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension

text: BACKGROUND: Dicer is a 219-kDa protein that plays key roles in gene regulation, particularly as the ribonuclease III enzyme responsible for cleaving precursor miRNA substrates. Its enzymatic activity is highly regulated by protein factors, and this regulation can impact on the levels of miRNAs and modulate the behavior of a cell. To better understand the underlying mechanisms of regulation, detailed enzymatic and structural characterization of Dicer are needed. However, these types of studies generally require several milligrams of recombinant protein, and efficient preparation of such quantities of pure human Dicer remains a challenge. To prepare large quantities of human Dicer, we have optimized transfection in HEK293-6E cells grown in suspension and streamlined a purification procedure. RESULTS: Transfection conditions were first optimized to achieve expression levels between 10 and 18 mg of recombinant Dicer per liter of culture. A three-step purification protocol was then developed that yields 4–9 mg of purified Dicer per liter of culture in a single day. From SEC-MALS/RI analysis and negative stain TEM, we confirmed that the purified protein is monomerically pure ( ≥ 98%) and folds with the characteristic L-shape geometry. Using an electrophoretic mobility shift assay, a dissociation constant (K(d)) of 5 nM was measured for Dicer binding to pre-let-7a-1, in agreement with previous reports. However, when probing the cleavage activity of Dicer for pre-let-7a-1, we measured k(cat) (7.2 ± 0.5 min(− 1)) and K(M) (1.2 ± 0.3 μM) values that are much higher than previously reported due to experimental conditions that better respect the steady-state assumption. CONCLUSIONS: The expression and purification protocols described here provide high yields of monomerically pure and active human Dicer. Cleavage studies of a pre-let-7 substrate with this purified Dicer reveal higher k(cat) and K(M) values than previously reported and support the current view that conformational changes are associated with substrate binding. Large quantities of highly pure Dicer will be valuable for future biochemical, biophysical and structural investigations of this key protein of the miRNA pathway. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12896-018-0485-3) contains supplementary material, which is available to authorized users.


title: Essential Oils as Antiviral Agents, Potential of Essential Oils to Treat SARS-CoV-2 Infection: An In-Silico Investigation

text: Essential oils have shown promise as antiviral agents against several pathogenic viruses. In this work we hypothesized that essential oil components may interact with key protein targets of the 2019 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A molecular docking analysis was carried out using 171 essential oil components with SARS-CoV-2 main protease (SARS-CoV-2 M(pro)), SARS-CoV-2 endoribonucleoase (SARS-CoV-2 Nsp15/NendoU), SARS-CoV-2 ADP-ribose-1″-phosphatase (SARS-CoV-2 ADRP), SARS-CoV-2 RNA-dependent RNA polymerase (SARS-CoV-2 RdRp), the binding domain of the SARS-CoV-2 spike protein (SARS-CoV-2 rS), and human angiotensin−converting enzyme (hACE2). The compound with the best normalized docking score to SARS-CoV-2 M(pro) was the sesquiterpene hydrocarbon (E)-β-farnesene. The best docking ligands for SARS−CoV Nsp15/NendoU were (E,E)-α-farnesene, (E)-β-farnesene, and (E,E)−farnesol. (E,E)−Farnesol showed the most exothermic docking to SARS-CoV-2 ADRP. Unfortunately, the docking energies of (E,E)−α-farnesene, (E)-β-farnesene, and (E,E)−farnesol with SARS-CoV-2 targets were relatively weak compared to docking energies with other proteins and are, therefore, unlikely to interact with the virus targets. However, essential oil components may act synergistically, essential oils may potentiate other antiviral agents, or they may provide some relief of COVID-19 symptoms.


title: Inflammatory and Coagulative Considerations for the Management of Orthopaedic Trauma Patients With COVID-19: A Review of the Current Evidence and Our Surgical Experience

text: Mounting evidence suggests that the pathogenesis of coronavirus disease 2019 (COVID-19) involves a hyperinflammatory response predisposing patients to thromboembolic disease and acute respiratory distress. In the setting of severe blunt trauma, damaged tissues induce a local and systemic inflammatory response through similar pathways to COVID-19. As such, patients with COVID-19 sustaining orthopaedic trauma injuries may have an amplified response to the traumatic insult because of their baseline hyperinflammatory and hypercoagulable states. These patients may have compromised physiological reserve to withstand the insult of surgical intervention before reaching clinical instability. In this article, we review the current evidence regarding pathogenesis of COVID-19 and its implications on the management of orthopaedic trauma patients by discussing a case and the most recent literature.


title: Diagnostic utility of a direct immunofluorescence test to detect feline coronavirus antigen in macrophages in effusive feline infectious peritonitis

text: Abstract The antemortem diagnosis of feline infectious peritonitis (FIP) remains challenging in clinical practice, since current testing methods have suboptimal diagnostic accuracy. Immunohistochemical testing of biopsy specimens and postmortem examination are the standard diagnostic methods, although direct immunofluorescence (DIF) testing to detect feline coronavirus in macrophages in effusion specimens has been reported to have 100% specificity and has been recommended as an antemortem confirmatory test. The aim of this study was to compare the results of DIF testing in antemortem feline effusions with postmortem results using field samples. Effusion specimens were collected antemortem from 17 cats and tested by DIF, followed by postmortem examination. Histopathological examination of specimens collected at postmortem confirmed FIP in 10/17 cases and ruled out FIP out in 7/17 cases. Antemortem DIF testing was positive in all 10 cases confirmed as FIP at postmortem examination. In the seven cats where FIP was ruled out at postmortem examination, DIF was negative in five cases and positive in the remaining two cases. The calculated sensitivity of DIF testing was 100% and the specificity was 71.4%. Duplicate effusion specimens from eight cats that were initially DIF positive were stored refrigerated (4°C) or at room temperature (22–25°C) and subjected to serial DIF testing to determine the duration of positive results. DIF-positive specimens stored at both temperatures retained their positive status for at least 2days.


title: The Nucleocapsid Protein of Coronavirus Infectious Bronchitis Virus: Crystal Structure of Its N-Terminal Domain and Multimerization Properties

text: Summary The coronavirus nucleocapsid (N) protein packages viral genomic RNA into a ribonucleoprotein complex. Interactions between N proteins and RNA are thus crucial for the assembly of infectious virus particles. The 45 kDa recombinant nucleocapsid N protein of coronavirus infectious bronchitis virus (IBV) is highly sensitive to proteolysis. We obtained a stable fragment of 14.7 kDa spanning its N-terminal residues 29–160 (IBV-N29-160). Like the N-terminal RNA binding domain (SARS-N45-181) of the severe acute respiratory syndrome virus (SARS-CoV) N protein, the crystal structure of the IBV-N29-160 fragment at 1.85 Å resolution reveals a protein core composed of a five-stranded antiparallel β sheet with a positively charged β hairpin extension and a hydrophobic platform that are probably involved in RNA binding. Crosslinking studies demonstrate the formation of dimers, tetramers, and higher multimers of IBV-N. A model for coronavirus shell formation is proposed in which dimerization of the C-terminal domain of IBV-N leads to oligomerization of the IBV-nucleocapsid protein and viral RNA condensation.


title: Comparison of Hospitalized Patients with pneumonia caused by COVID-19 and influenza A in children under 5 years

text: Abstract Background Since the outbreak of Coronavirus Disease 2019 (COVID-19) in Wuhan, considerable attention has been paid on its epidemiology and clinical characteristics in children patients. However, it is also crucial for clinicians to differentiate COVID-19 from other respiratory infectious diseases, such as influenza viruses. Methods This was a retrospective study. Two group of COVID-19 patients (n=57) and influenza A patients (n=59) were enrolled. We analyzed and compared their clinical manifestations, imaging characteristics and treatments. Results The proportions of cough (70.2%), fever (54.4%) and gastrointestinal symptoms (14.1%) in COVID-19 patients were lower than those of influenza A patients (98.3%, P<0.001; 84.7%, P<0.001; and 35.6%, P=0.007; respectively). In addition, COVID-19 patients showed significantly lower levels of leukocytes (7.87 vs. 9.89×109/L, P=0.027), neutrophils (2.43 vs. 5.16×109/L, P<0.001), C-reactive protein (CRP; 3.7 vs. 15.1mg/L, P=0.001) and procalcitonin (PCT; 0.09 vs. 0.68mm/h, P<0.001), while lymphocyte levels (4.58 vs. 3.56×109/L; P=0.006) were significantly higher compared with influenza A patients. In terms of CT imaging, ground-glass opacification in chest CT was more common in COVID-19 patients than in influenza A patients (42.1% vs. 15%, P=0.032). In contrast, consolidation was more common in influenza A patients (25%) than that in COVID-19 patients (5.2%, P=0.025). Conclusion The clinical manifestations and laboratory tests of COVID-19 children are milder than those of influenza A children under 5 years. Additionally, imaging results more commonly presented as ground-glass opacities in COVID-19 patients.