title: Onset and duration of protective immunity against clinical disease and renal carriage in dogs provided by a bi-valent inactivated leptospirosis vaccine

text: Abstract Protection against clinical disease and prevention of the renal carrier state remain the key objectives of vaccination against leptospirosis in the dog. In the present paper, groups of dogs were vaccinated twice with a commercial bacterin (EURICAN® L) containing Leptospira interrogans serovars icterohaemorrhagiae and canicola and challenged with heterologous representatives of both serovars at 2 weeks (onset of immunity) or 14 months (duration of immunity) after the second vaccination. Control dogs were not vaccinated against leptospirosis and kept with the vaccinated dogs. The challenges, irrespective of the serovar, reliably produced clinical signs consistent with Leptospira infection in the control pups with up to 60% mortality. As expected clinical disease in the adult controls was less severe, but we were able to induce morbidity and mortality as well. Under these extreme challenge conditions, clinical signs in the vaccinated dogs were rare, and when observed, mild and transient in nature. Following experimental infection, 100% of the control pups and 83% of the adult controls became renal carriers. Despite the heavy challenges, none of the 18 vaccinated puppies (onset of immunity studies) and only 2 out of the 16 vaccinated adult dogs (duration of immunity studies) developed a renal carrier state. These results show that a primary course of two doses of EURICAN® L provided quick onset and long-term protection against both clinical leptospirosis and the renal carrier stage. This vaccine should provide veterinarians with a powerful tool to prevent clinical disease in dogs and zoonotic transmission of leptospirosis to humans.


title: Characterization and application of monoclonal antibodies against N protein of SARS-coronavirus

text: Abstract Severe acute respiratory syndrome-coronavirus (SARS-CoV) causes an infectious disease through respiratory route. Diagnosing the disease effectively and accurately at early stage is essential for preventing the disease transmission and performing antiviral treatment. In this study, we raised monoclonal antibodies (mAbs) against the nucleocapsid (N) protein of SARS-CoV and mapped epitopes by using different truncated N protein fragments. The mapping of those epitopes was valuable for constructing pair-Abs used in serological diagnosis. The results showed that all of the six raised mAbs were divided into two groups recognizing the region of amino acids 249–317 (A group) or 317–395 (B group). This region spanning amino acids 249–395 contains predominant B cell epitopes located at the C-terminus of N protein. One pair-Abs, consisting of N protein-specific rabbit polyclonal antibody and SARS-CoV N protein-specific mAb, was selected to construct a sandwich ELISA-kit. The kit was able to specifically detect SARS-CoV N proteins in serum samples.


title: Association between secondary thrombocytosis and viral respiratory tract infections in children

text: Secondary thrombocytosis (ST) is frequently observed in children with a variety of clinical conditions. The leading cause of ST is respiratory tract infection (RTI) in children. Nasopharyngeal aspirate samples were collected and assessed for common respiratory viruses. The relationships between virus infections and secondary thrombocytosis were analyzed retrospectively. The blood platelet count and the presence of respiratory viruses were determined for 3156 RTI patients, and 817 (25.9%) cases with platelet ≥500 × 10(9)/L were considered as the thrombocytosis group. Compared with the normal group, the detection rates of respiratory syncytial virus (RSV) and human rhinovirus (HRV) were significantly higher in the thrombocytosis group (P = 0.017 and 0.042, respectively). HRV single infection was a risk factor associated with thrombocytosis [odds ratio (OR) = 1.560, 95% confidence interval (CI) = 1.108–2.197]. Furthermore, ST was more likely to occur in younger patients who had clinical manifestations of wheezing and dyspnea and who had been diagnosed with bronchiolitis. Furthermore, the course of disease lasted longer in these patients. ST is associated with viral respiratory tract infections, especially RSV and HRV infections. HRV single infection is a risk factor associated with thrombocytosis.


title: Frontiers in climate change–disease research

text: The notion that climate change will generally increase human and wildlife diseases has garnered considerable public attention, but remains controversial and seems inconsistent with the expectation that climate change will also cause parasite extinctions. In this review, we highlight the frontiers in climate change–infectious disease research by reviewing knowledge gaps that make this controversy difficult to resolve. We suggest that forecasts of climate-change impacts on disease can be improved by more interdisciplinary collaborations, better linking of data and models, addressing confounding variables and context dependencies, and applying metabolic theory to host–parasite systems with consideration of community-level interactions and functional traits. Finally, although we emphasize host–parasite interactions, we also highlight the applicability of these points to climate-change effects on species interactions in general.


title: Neonatal diarrhea in llamas and alpacas

text: Abstract Diarrhea is an important cause of morbidity in neonatal llamas and alpacas. Diarrhea may be multifactorial in etiology including management and nutritional factors as well as a variety of pathogens. Most of the pathogens involved affect other livestock species and some have host-adapted strains. However, the clinical signs, their expected severity and age of onset of disease varies between species in some cases. The most common pathogens causing diarrhea in neonatal camelids are coronavirus, Escherichia coli (E. coli), Cryptosporidium spp., Giardia spp. and coccidia. The purpose of this paper is to review the available literature on neonatal diarrhea in camelids and to present clinical data from 55 cases seen at The Ohio State University.


title: Preventive and therapeutic efficacy of halofuginone-lactate against Cryptosporidium parvum in spontaneously infected calves: A centralised, randomised, double-blind, placebo-controlled study

text: Abstract The preventive and therapeutic efficacy of halofuginone-lactate (HFL) against Cryptosporidium parvum was evaluated in a study conducted from November 2004 to March 2005 on a dairy farm in Central Bohemia, Czech Republic, using 260 spontaneously infected calves. HFL (0.1mg/kg/day) was administered orally for 7 days to 1-day-old and 8-day-old calves, respectively. In both treated groups the drug significantly, and in almost the same manner, decreased the intensity of diarrhoea (P <0.001) and faecal oocyst count (P <0.001) when compared to corresponding placebo groups. The only difference between both treated groups was the time of onset of symptoms of the infection. Over time, the clinical pattern of cryptosporidiosis in the animals treated at 8–14 days of age was similar to that seen in the groups receiving the placebo. In contrast, infection in the preventively treated group peaked about 10 days later but with the same intensity. The results of this study confirm the anticryptosporidial activity of HFL in calves, but show that the outcome of infection following preventive treatment is comparable to that observed in calves treated after the onset of symptoms.


title: Vaccination of influenza a virus decreases transmission rates in pigs

text: Limited information is available on the transmission and spread of influenza virus in pig populations with differing immune statuses. In this study we assessed differences in transmission patterns and quantified the spread of a triple reassortant H1N1 influenza virus in naïve and vaccinated pig populations by estimating the reproduction ratio (R) of infection (i.e. the number of secondary infections caused by an infectious individual) using a deterministic Susceptible-Infectious-Recovered (SIR) model, fitted on experimental data. One hundred and ten pigs were distributed in ten isolated rooms as follows: (i) non-vaccinated (NV), (ii) vaccinated with a heterologous vaccine (HE), and (iii) vaccinated with a homologous inactivated vaccine (HO). The study was run with multiple replicates and for each replicate, an infected non-vaccinated pig was placed with 10 contact pigs for two weeks and transmission of influenza evaluated daily by analyzing individual nasal swabs by RT-PCR. A statistically significant difference between R estimates was observed between vaccinated and non-vaccinated pigs (p < 0.05). A statistically significant reduction in transmission was observed in the vaccinated groups where R (95%CI) was 1 (0.39-2.09) and 0 for the HE and the HO groups respectively, compared to an R(o )value of 10.66 (6.57-16.46) in NV pigs (p < 0.05). Transmission in the HE group was delayed and variable when compared to the NV group and transmission could not be detected in the HO group. Results from this study indicate that influenza vaccines can be used to decrease susceptibility to influenza infection and decrease influenza transmission.


title: Relationship of α2‐Macroglobulin with Steroid‐Induced Femoral Head Necrosis: A Chinese Population‐Based Association Study in Southeast China

text: OBJECTIVE: The present study aimed to identify the relationship of α‐2‐macroglobulin and microvascular vessel pathology with steroid‐induced femoral head necrosis in the Southeast Chinese population. METHODS: This study enrolled 40 patients diagnosed with steroid‐induced necrosis of the femoral head. Patients had various stages of femoral head necrosis. The differential expression of serum proteins and mRNA from patients with steroid‐induced necrosis of the femoral head (SINFH) and healthy volunteers was analyzed by western blot and quantitative polymerase chain reaction (QT‐PCR). The pathological change in osteocyte necrosis was indicated by hematoxylin and eosin stain and immunohistochemistry. RESULTS: Hematoxylin and eosin stain showed histopathology changes in the necrotic area of patients with steroid‐induced INFH: bone trabeculae were fewer and thinner, became broken, fragmented and structurally disordered; intraosseous adipose cells became enlarged; the arrangement of the osteoblasts became irregular; and vacant bone lacunae increased. QT‐PCR showed significantly lower levels of α‐2‐macroglobulin in the serum of patients with SINFH than in controls (P < 0.05). Immunohistochemical staining and western blotting demonstrated that the expression of α‐2‐macroglobulin was significantly decreased in the necrotic area of SINFH patients (P < 0.05). CONCLUSION: The α‐2‐macroglobulin may be associated with the pathology of SINFH. The multiple pathological reactions occur in SINFH and α‐2‐macroglobulin may serve as a potential biomarker for the diagnosis of SINFH or a promising therapeutic target.


Social capital and sleep quality in those who self-isolate


Sentiment Analysis of Nationwide Lockdown due to COVID 19 Outbreak: Evidence from India


title: Experimental investigation of backpulse and backblow cleaning of nanofiber filter loaded with nano-aerosols

text: Abstract Nanofibrous filter have been proven effective to remove nano-aerosols with size less than 100nm. Cleaning is required after long-term use; however, very little has been published on the subject. An experimental investigation has been launched to determine backpulse, backblow and combined backpulse–backblow on cleaning of a loaded nanofiber filter. Nylon 6 nanofiber filters were loaded with polydispersed NaCl particles, 60%<100nm and 90%<160nm, generated from an aerosol generator. Air jets in form of backpulse, backblow and their combined mode were used to clean a loaded filter. During cleaning, the filter cake was removed first for which the pressure drop across the loaded filter decreased rapidly, followed by loosely attached aerosols in the filter being removed with finite pressure drop reduction at a reasonable rate, ending in the final stage for which much lesser aerosols were being removed. Ultimately, the filter reached a residual pressure drop which was higher than that of the initial clean filter indicating residual aerosols were trapped both in the cake heel and filter. Backpulse has been found to be more effective in removing the cake from the filter surface, whereas backblow provides an added advantage of removing by convection of the detached aerosols away from the filter preventing recapture. The synergistic combination of backpulse–backblow provides the best cleaning performance of a nanofibrous filter loaded with nano-aerosols.


title: Subtotal laparoscopic cholecystectomy for gangrenous gallbladder during recovery from COVID-19 pneumonia

text: INTRODUCTION: Management of acute abdomen during COVID-19 pandemic may be challenging. Presentation of case. A 42-year old man was hospitalized for Covid-19 pneumonia. Fever, respiratory symptoms and hypoxemia significantly improved over the next 2 weeks, but the patient developed abdominal pain, nausea, and low-grade fever. Computed tomography scan revealed absence of contrast enhancement of gallbladder wall and a micro-perforation of the fundus. At laparoscopy, gallbladder gangrene was confirmed and a subtotal cholecystectomy performed. Special precautions were adopted for patient transportation from the ward to a dedicated operating room, and two teams with adequate personal protective equipment took charge of the procedure. The patient was discharged home on postoperative day 7 under protective lockdown measures for 2 weeks. DISCUSSION: The pathogenesis of acute acalcolous gangrenous cholecystitis is multifactorial. It is unknown whether a prothrombotic state induced by COVID-19 contributes to wall ischemia and perforation. Percutaneous cholecystostomy should be avoided in patients with gallbladder gangrene. Contraindications to laparoscopy are not evidence-based since aerosolization is produced during both open and laparoscopic surgical procedures. However, personal protective equipment is key for prevention CONCLUSION: Early diagnosis and surgical therapy are critical in patients with gangrenous cholecystitis. Subtotal laparoscopic cholecystectomy for gangrenous gallbladder is safe and effective.


title: Immunomodulatory therapy for the management of severe COVID-19. Beyond the anti-viral therapy: A comprehensive review

text: Severe Acute Respiratory Syndrome related to Coronavirus-2 (SARS-CoV-2), coronavirus disease-2019 (COVID-19) may cause severe illness in 20% of patients. This may be in part due to an uncontrolled immune-response to SARS-CoV-2 infection triggering a systemic hyperinflammatory response, the so-called “cytokine storm”. The reduction of this inflammatory immune-response could be considered as a potential therapeutic target against severe COVID-19. The relationship between inflammation and clot activation must also be considered. Furthermore, we must keep in mind that currently, no specific antiviral treatment is available for SARS-CoV-2. While moderate-severe forms need in-hospital surveillance plus antivirals and/or hydroxychloroquine; in severe and life-threating subsets a high intensity anti-inflammatory and immunomodulatory therapy could be a therapeutic option. However, right data on the effectiveness of different immunomodulating drugs are scarce. Herein, we discuss the pathogenesis and the possible role played by drugs such as: antimalarials, anti-IL6, anti-IL-1, calcineurin and JAK inhibitors, corticosteroids, immunoglobulins, heparins, angiotensin-converting enzyme agonists and statins in severe COVID-19.


title: Comparison of the Panther Fusion and a laboratory-developed test targeting the envelope gene for detection of SARS-CoV-2

text: Abstract Background Numerous nucleic acid amplification assays have recently received emergency use authorization (EUA) for the diagnosis of SARS-CoV-2 infection, and there is a need to assess their test performance relative to one another. Objectives The aim of this study was to compare the test performance of the Hologic Panther Fusion SARS-CoV-2 assay targeting two regions of open reading frame 1ab (ORF1ab) to a high complexity molecular-based, laboratory-developed EUA from Stanford Health Care (SHC) targeting the SARS-CoV-2 envelope (E) gene. Study design We performed a diagnostic comparison study by testing nasopharyngeal samples on the two assays. Assay agreement was assessed by overall percent agreement and Cohen’s kappa coefficient. Results A total of 184 nasopharyngeal samples were tested using the two assays, of which 180 showed valid results and were included for the comparative analysis. Overall percent agreement between the assays was 98.3 % (95 % confidence interval (CI) 95.2–99.7) and kappa coefficient was 0.97 (95 % CI 0.93–1.0). One sample was detected on the SHC laboratory developed test (LDT) and not on the Panther Fusion, and had a Ct of 35.9. Conversely, 2 samples were detected on the Panther Fusion and not on the LDT, and had Ct values of 37.2 and 36.6. Conclusion The Panther Fusion SARS-CoV-2 assay and the SHC LDT perform similarly on clinical nasopharyngeal swab specimens. Other considerations, including reagent availability, turnaround time, labor requirements, cost and instrument throughput should guide the decision of which assay to perform.


title: Research and Development on Therapeutic Agents and Vaccines for COVID-19 and Related Human Coronavirus Diseases

text: Since the outbreak of the novel coronavirus disease COVID-19, caused by the SARS-CoV-2 virus, this disease has spread rapidly around the globe. Considering the potential threat of a pandemic, scientists and physicians have been racing to understand this new virus and the pathophysiology of this disease to uncover possible treatment regimens and discover effective therapeutic agents and vaccines. To support the current research and development, CAS has produced a special report to provide an overview of published scientific information with an emphasis on patents in the CAS content collection. It highlights antiviral strategies involving small molecules and biologics targeting complex molecular interactions involved in coronavirus infection and replication. The drug-repurposing effort documented herein focuses primarily on agents known to be effective against other RNA viruses including SARS-CoV and MERS-CoV. The patent analysis of coronavirusrelated biologics includes therapeutic antibodies, cytokines, and nucleic acid-based therapies targeting virus gene expression as well as various types of vaccines. More than 500 patents disclose methodologies of these four biologics with the potential for treating and preventing coronavirus infections, which may be applicable to COVID-19. The information included in this report provides a strong intellectual groundwork for the ongoing development of therapeutic agents and vaccines.


title: COVID-19 and molecular mimicry: the Columbus’ egg?

text: • Recently, this Journal published a report about Guillain-Barré syndrome associated with COVID-19 infection. • Guillain-Barré syndrome can be due to molecular mimicry phenomena. • Molecular mimicry had already been described in another SARS. • It could explain the autoimmune signs and symptoms that some patients affected by SARS-CoV-2 can experience.


Ecosystem approaches to the risk for schizophrenia


title: Radioactive and enzymatic cloned cDNA probes for bovine enteric coronavirus detection by molecular hybridization

text: Genomic RNA of F15 strain bovine enteric coronavirus (BECV) was cloned inE. coli. Three clones (174, 160, PG 78), selected in the cDNA library, including a large portion of the nucleocapsid (N), matrix (M) and peplomeric (S) protein genes, were used as probes for a slot blot hybridization assay. Two probe labelling techniques were compared, radiolabelling with(32)P and enzymatic labelling through covalent linkage to peroxidase and chemiluminescence detection. The radioactive probe 174 detected as little as 1 to 3 pg of viral RNA, while the less sensitive enzymatic probe could not reveal more than 100 pg of RNA. No significant detection amplification was achieved when a mixture of the three probes was used. Probe 174 allowed specific identification for BECV. No hybridization was noticed either with rotaviruses or even with other antigenically unrelated members of the familyCoronaviridae such as transmissible gastroenteritis virus. The test proved valid for detection of BECV in the supernatant of infected HRT-18 cells: genomic RNA could be detected after direct spotting of samples, but prior nucleic acid extraction after proteinase K treatment improved virus detection. BECV diagnosis in faecal samples using enzymatic probe was compared with conventional diagnostic methods.


Practical tips for using masks in the COVID‐19 pandemic


title: Bronchiolitis: Analysis of 10 consecutive epidemic seasons

text: Bronchiolitis is the leading cause of hospitalization in infants under 12 months. Our aims were to analyze epidemiological characteristics of infants with bronchiolitis over 10 consecutive seasons and to evaluate whether there are any clinical differences between infants hospitalized for bronchiolitis during epidemic peak months and infants in non‐peak months. We enrolled consecutive enrolled 723 previously healthy term infants hospitalized at the Paediatric Emergency Department, “Sapienza” University of Rome over the period 2004–2014. Fourteen respiratory viruses were detected from nasopharyngeal aspirates by molecular methods. Clinical and demographic data were extracted from clinical charts. Viruses were detected in 351 infants (48.5%): RSV in 234 (32.4%), RV in 44 (6.1%), hBoV in 11 (1.5%), hMPV in 12 (1.6%), co‐infections in 39 (5.4%), and other viruses in 11 (1.5%). Analyzing the 10 epidemic seasons, we found higher incidence for bronchiolitis every 4 years with a peak during the months December–January. Infants hospitalized during peak months had lower family history for asthma (P = 0.003), more smoking mothers during pregnancy (P = 0.036), were slightly higher breastfed (0.056), had lower number of blood eosinophils (P = 0.015) and had a higher clinical severity score (P = 0.017). RSV was detected mostly during peak months, while RV was equally distributed during the seasons. We found some variations in bronchiolitis incidence during epidemics, and discriminative characteristics in infants hospitalized for bronchiolitis during peak months and in non‐peak months, that might reflect two different populations of children. Pediatr Pulmonol. 2016;51:1330–1335. © 2016 Wiley Periodicals, Inc.


title: Perceptions of hand hygiene practices in China

text: Summary Hand hygiene is considered one of the most important infection control measures for preventing healthcare-associated infections. However, compliance rates with recommended hand hygiene practices in hospitals remain low. Previous literature on ways to improve hand hygiene practices has focused on the USA and Europe, whereas studies from developing countries are less common. In this study, we sought to identify common issues and potential strategies for improving hand hygiene practices in hospitals in China. We used a qualitative survey design based on in-depth interviews with 25 key hospital and public health staff in eight hospitals selected by the Chinese Ministry of Health. We found that hospital workers viewed hand hygiene as paramount to effective infection control and had adequate knowledge about proper hand hygiene practices. Despite these positive attitudes and adequate knowledge, critical challenges to improving rates of proper hand hygiene practices were identified. These included lack of needed resources, limited organisational authority of hospital infection control departments, and ineffective use of data monitoring and feedback to motivate improvements. Our study suggests that a pivotal issue for improving hand hygiene practice in China is providing infection control departments adequate attention, priority, and influence within the hospital, with a clear line of authority to senior management. Elevating the place of infection control on the hospital organisational chart and changing the paradigm of surveillance to continuous monitoring and effective data feedback are central to achieving improved hand hygiene practices and quality of care.


Let's deconfine carefully for not having to reconfine in a hurry


title: Tumoricidal activity of human dendritic cells

text: Dendritic cells (DCs) are a family of professional antigen-presenting cells (APCs) that are able to initiate innate and adaptive immune responses against pathogens and tumor cells. The DC family is heterogeneous and is classically divided into two main subsets, each with its unique phenotypic and functional characteristics: myeloid DCs (mDCs) and plasmacytoid DCs (pDCs). Recent results have provided intriguing evidence that both DC subsets can also function as direct cytotoxic effector cells; in particular, against cancer cells. In this review, we delve into this understudied function of human DCs and discuss why these so-called killer DCs might become important tools in future cancer immunotherapies.


title: Low-cost, easy-to-build non-invasive pressure support ventilator for under-resourced regions: open source hardware description, performance and feasibility testing

text: AIM: Current pricing of commercial mechanical ventilators in low/middle-income countries (LMICs) markedly restricts their availability, and consequently a considerable number of patients with acute/chronic respiratory failure cannot be adequately treated. Our aim was to design and test an affordable and easy-to-build non-invasive bilevel pressure ventilator to allow reducing the serious shortage of ventilators in LMICs. METHODS: The ventilator was built using off-the-shelf materials available via e-commerce and was based on a high-pressure blower, two pressure transducers and an Arduino Nano controller with a digital display (total retail cost <75 US$), with construction details open source provided for free replication. The ventilator was evaluated (and compared with a commercially available device (Lumis-150, Resmed): a) in the bench using an actively breathing patient simulator mimicking a range of obstructive/restrictive disease and b) in 12 healthy volunteers wearing a high airway resistance and thoracic/abdominal bands to mimic obstructive/restrictive patients. RESULTS: The designed ventilator provided inspiratory/expiratory pressures up to 20/10 cmH(2)O, respectively, with no faulty triggering or cycling both in the bench test and in volunteers. Breathing difficulty score rated (1–10 scale) by the loaded breathing subjects was significantly (p<0.005) decreased from 5.45±1.68 without support to 2.83±1.66 when using the prototype ventilator, which showed no difference with the commercial device (2.80±1.48; p=1.000). CONCLUSION: The low-cost, easy-to-build non-invasive ventilator performs similarly as a high-quality commercial device, with its open-source hardware description, will allow for free replication and use in LMICs, facilitating application of this life-saving therapy to patients who otherwise could not be treated.


title: Exploring the ways in which COVID‐19 and lockdown has affected the lives of adult patients with anorexia nervosa and their carers

text: OBJECTIVE: This qualitative study explores the ways in which the coronavirus disease 2019 (COVID‐19) pandemic and associated lockdown measures have affected the lives of adult patients with anorexia nervosa (AN) and their carers. METHOD: Semi‐structured interviews were conducted with patients with AN (n = 21) and carers (n = 28) from the start of UK Government imposed lockdown. Data related directly to the impact of lockdown and COVID‐19 were analysed using thematic analysis. RESULTS: Four broad themes were identified for patients and carers separately. Patients experienced: 1. reduced access to eating disorder (ED) services; 2. disruption to routine and activities in the community; 3. heightened psychological distress and ED symptoms; 4. increased attempts at self‐management in recovery. Carer themes included: 1. concern over provision of professional support for patients; 2. increased practical demands placed on carers in lockdown; 3. managing new challenges around patient wellbeing; 4. new opportunities. CONCLUSIONS: Reduced access to ED services, loss of routine and heightened anxieties and ED symptoms resulting from COVID‐19 and lockdown measures presented challenges for patients and carers. Increased remote support by ED services enabled the continuation of treatment and self‐management resources and strategies promoted self‐efficacy in both groups.


title: Rats and Mice

text: This chapter provides an overview of the natural history, anatomy, physiology, clinical examination, common diseases, and treatment of rats (Rattus norvegicus) and mice (Mus musculus) in the context of veterinary medicine. Guidelines for the care and feeding of rats and mice can provide owners with information to help prevent disease in their pets. Useful techniques for restraint, clinical examination, and diagnostic sample collection are provided to aid veterinarians in thorough evaluation of these small rodents. Common diseases and treatments are discussed separately for each species, organized by organ system. Zoonotic diseases are also discussed to provide guidance for rat and mouse pet owners.


title: Die Corona-Pandemie 2020 – über eine allumfassende Prävention hinaus

text: This paper is part of Forum COVID-19: Perspectives in the Humanities and Social Sciences. The Spanish Flu 1918–1920 caused between 50 and 100 million deaths. Despite this, West German officials ignored the pandemics of 1957/1958 and 1968–1970. Patient perseverance seems to be an appropriate label for the lack of any action. The appearance of new viruses had a massive impact on the discourse concerning pandemics: “patient perseverance” became “omnipresent prevention.” The actual measures against SARS-CoV‑2 exceed the “omnipresent prevention” used during the 2009 swine flu pandemic by far.


title: Development and validation of a real-time PCR assay for specific and sensitive detection of canid herpesvirus 1

text: Abstract A TaqMan-based real-time PCR assay targeting the glycoprotein B-encoding gene was developed for diagnosis of canid herpesvirus 1 (CHV-1) infection. The established assay was highly specific, since no cross-reactions were observed with other canine DNA viruses, including canine parvovirus type 2, canine minute virus, or canine adenovirus types 1 and 2. The detection limit was 101 and 1.20×101 DNA copies per 10μl−1 of template for standard DNA and a CHV-1-positive kidney sample, respectively: about 1-log higher than a gel-based PCR assay targeting the thymidine kinase gene. The assay was also reproducible, as shown by satisfactory low intra-assay and inter-assay coefficients of variation. CHV-1 isolates of different geographical origins were recognised by the TaqMan assay. Tissues and clinical samples collected from three pups which died of CHV-1 neonatal infection were also tested, displaying a wide distribution of CHV-l DNA in their organs. Unlike other CHV-1-specific diagnostic methods, this quantitative assay permits simultaneous detection and quantitation of CHV-1 DNA in a wide range of canine tissues and body fluids, thus providing a useful tool for confirmation of a clinical diagnosis, for the study of viral pathogenesis and for evaluation of the efficacy of vaccines and antiviral drugs.


title: Eating habits and lifestyle changes during COVID-19 lockdown: an Italian survey

text: BACKGROUND: On December 12th 2019, a new coronavirus (SARS-Cov2) emerged in Wuhan, China, sparking a pandemic of acute respiratory syndrome in humans (COVID-19). On the 24th of April 2020, the number of COVID-19 deaths in the world, according to the COVID-Case Tracker by Johns Hopkins University, was 195,313, and the number of COVID-19 confirmed cases was 2,783,512. The COVID-19 pandemic represents a massive impact on human health, causing sudden lifestyle changes, through social distancing and isolation at home, with social and economic consequences. Optimizing public health during this pandemic requires not only knowledge from the medical and biological sciences, but also of all human sciences related to lifestyle, social and behavioural studies, including dietary habits and lifestyle. METHODS: Our study aimed to investigate the immediate impact of the COVID-19 pandemic on eating habits and lifestyle changes among the Italian population aged ≥ 12 years. The study comprised a structured questionnaire packet that inquired demographic information (age, gender, place of residence, current employment); anthropometric data (reported weight and height); dietary habits information (adherence to the Mediterranean diet, daily intake of certain foods, food frequency, and number of meals/day); lifestyle habits information (grocery shopping, habit of smoking, sleep quality and physical activity). The survey was conducted from the 5th to the 24th of April 2020. RESULTS: A total of 3533 respondents have been included in the study, aged between 12 and 86 years (76.1% females). The perception of weight gain was observed in 48.6% of the population; 3.3% of smokers decided to quit smoking; a slight increased physical activity has been reported, especially for bodyweight training, in 38.3% of respondents; the population group aged 18–30 years resulted in having a higher adherence to the Mediterranean diet when compared to the younger and the elderly population (p < 0.001; p < 0.001, respectively); 15% of respondents turned to farmers or organic, purchasing fruits and vegetables, especially in the North and Center of Italy, where BMI values were lower. CONCLUSIONS: In this study, we have provided for the first time data on the Italian population lifestyle, eating habits and adherence to the Mediterranean Diet pattern during the COVID-19 lockdown. However, as the COVID-19 pandemic is ongoing, our data need to be confirmed and investigated in future more extensive population studies.


title: Age, source, and future risk of COVID-19 infections in two settings of Hong Kong and Singapore

text: OBJECTIVE: To explore and compare the age, source and future risk of COVID-19 infection in Hong Kong SAR China and Singapore as of March 5, 2020. RESULTS: We find significant difference in age patterns of confirmed cases in these 2 localities early in the pandemic. CONCLUSION: We highlight the potential importance of population age structure in confirmed cases, which should be considered in evaluation of the effectiveness of control effort in different localities.


title: Specific antibody secreting cells from chickens can be detected by three days and memory B cells by three weeks post-infection with the avian respiratory coronavirus

text: Infectious bronchitis virus (IBV), the first coronavirus described, has been a continuing problem in poultry for more than 70 years. IBV, causing a highly contagious respiratory disease in chickens, resembles the recently described severe acute respiratory syndrome virus in pathogenesis and genome organization. While previous studies demonstrated that effector and memory CD8(+) T lymphocytes are critical in controlling acute IBV infection and disease in chickens, here chicken anti-IBV antibody (IgG) secreting cells (ASC) in both peripheral blood mononuclear cells (PBMC) and spleens collected following IBV Gray infection were evaluated using an ELISPOT assay. The ASC in peripheral blood and spleens can be detected from 3 to 7 days post-infection (p.i.), which is 3–7 days earlier than anti-IBV IgG detected in the serum. The ASC frequency reached a maximum at 7–10 days p.i., and decreased more than 90% in the spleen and 70% in PBMC by 14 days p.i. The ASC levels in the PBMC then decreased gradually to 0.5 ASC/10(6) over the next 8 weeks. The higher concentration of about 20 ASC/10(6) cells in spleens may, at least partially, account for the presence of antibody in the serum although bone marrow ASC were not determined. In vitro stimulation of PBMC and splenocytes with IBV antigen demonstrated that memory B cells can be activated to secrete antibody by 3 weeks p.i. ELISPOT detection of primary B cells could be useful in the early detection of infection following infection with respiratory coronaviruses.


title: Chapter 22 Infrared Spectroscopic Technologies for the Quality Control of Herbal Medicines

text: Abstract The properties of herbal medicines are related to certain classes of compounds such as alkaloids, flavonoids, essential oils, and others. During the last four decades, near-infrared (NIR) spectroscopy (800–2500nm; 12.500–4.000/cm) has become one of the most attractive and widely used methods for analysis rather than traditionally applied separation methods (including liquid chromatography and electrophoresis) for the following reasons: NIR spectroscopy is a noninvasive analytical tool allowing a fast and simultaneous qualitative and quantitative characterization of herbal medicines and their constituents. Infrared imaging techniques are becoming more and more attractive to understand the function and the biochemical composition of plant tissue. In this contribution, the principle, technique, and methodology are described, followed by a discussion of quantitative and qualitative application possibilities. Finally, some hints to helpful regulatory issues are summarized.


title: Labour issues and COVID‐19

text: The COVID‐19 pandemic has prompted Canada and several other countries to impose an economic shutdown to prevent a deadly public health crisis from becoming much deadlier. In the agriculture and food sector, several hundred thousand restaurant workers have lost their jobs. The rise in unemployment, the closing of restaurants and schools, and social distancing have triggered demand reductions for certain commodities and foods and demand increases for others, bringing along changes in demand for inputs including labour. Canadian employers of temporary foreign workers (TFWs) are facing delays and additional constraints in recruiting, but so have US and European employers of TFWs. Rising food security concerns are making protectionist trade policies popular. Domestic and foreign firms may export less and do more foreign direct investment, inducing trade in jobs. This article is protected by copyright. All rights reserved


Coronavirus disease 2019 (COVID‐19): angiotensin‐converting enzyme inhibitors, angiotensin II receptor blockers and cardiovascular disease


title: Human bocavirus in the nasopharynx of otitis-prone children

text: Abstract Objectives Human bocavirus (HBoV) is frequently identified in children with respiratory tract infections, and its role in acute otitis media (AOM) has been suggested. The disease associations for the closely related bocaviruses HBoV2-4 remain unknown. Increasing evidence shows that probiotics may reduce the risk of AOM of viral origin. Objectives of the study was to examine the prevalence and persistence of bocaviruses in consecutive nasopharyngeal samples (NPS) of otitis-prone children, and whether an association exists between HBoV and the child's characteristics, respiratory symptoms, and AOM pathogens, and whether probiotics reduce the occurrence of HBoV. Methods In a double-blind, placebo-controlled, randomized, 6-month intervention study, 269 otitis-prone children (aged 9 months to 5.6 years), consumed daily either one capsule of probiotics (Lactobacillus rhamnosus GG, L. rhamnosus Lc705, Bifidobacterium breve 99 and Propionibacterium freudenreichii JS) or placebo. After a clinical examination and NPS collected at three-time points, the presence and persistence of HBoV1-4 DNA in NPS was determined by RT-qPCR at the baseline, after 3, and 6 months. Results A high load (>10,000copies/ml) of HBoV DNA was detected in 26 (17.1%) of 152 children, and 16 (10.5%) showed a prolonged presence of HBoV for at least 3 months. None had DNA of HBoV2-4. Higher number of siblings associated with increased HBoV prevalence (p =0.029). Prevalence or persistence of HBoV was not significantly associated with other characteristics, respiratory symptoms, or AOM pathogens. Probiotic intervention significantly reduced the number of HBoV DNA-positive samples (probiotic vs. placebo: 6.4% vs. 19.0%, OR=0.25, CI 95%=0.07–0.94, p =0.039). Conclusions HBoV, but not HBoV2-4, DNA occurs often in the nasopharynx of otitis-prone children, and may persist for 3–6 months. Probiotic treatment possibly reduced the presence of HBoV.


title: Correction to: COVID-19 pandemic: the 3R’s (reduce, refine, and replace) of personal protective equipment (PPE) sustainability

text: This article was updated to fix a typo in the title introduced during the production process.


title: Lung Ultrasound in COVID-19 Pneumonia: Correlations with Chest CT on Hospital admission

text: BACKGROUND: Lung ultrasound (LUS) is an accurate, safe, and cheap tool assisting in the diagnosis of several acute respiratory diseases. The diagnostic value of LUS in the workup of coronavirus disease-19 (COVID-19) in the hospital setting is still uncertain. OBJECTIVES: The aim of this observational study was to explore correlations of the LUS appearance of COVID-19-related pneumonia with CT findings. METHODS: Twenty-six patients (14 males, age 64 ± 16 years) urgently hospitalized for COVID-19 pneumonia, who underwent chest CT and bedside LUS on the day of admission, were enrolled in this observational study. CT images were reviewed by expert chest radiologists, who calculated a visual CT score based on extension and distribution of ground-glass opacities and consolidations. LUS was performed by clinicians with certified competency in thoracic ultrasonography, blind to CT findings, following a systematic approach recommended by ultrasound guidelines. LUS score was calculated according to presence, distribution, and severity of abnormalities. RESULTS: All participants had CT findings suggestive of bilateral COVID-19 pneumonia, with an average visual scoring of 43 ± 24%. LUS identified 4 different possible abnormalities, with bilateral distribution (average LUS score 15 ± 5): focal areas of nonconfluent B lines, diffuse confluent B lines, small subpleural microconsolidations with pleural line irregularities, and large parenchymal consolidations with air bronchograms. LUS score was significantly correlated with CT visual scoring (r = 0.65, p < 0.001) and oxygen saturation in room air (r = −0.66, p < 0.001). CONCLUSION: When integrated with clinical data, LUS could represent a valid diagnostic aid in patients with suspect COVID-19 pneumonia, which reflects CT findings.


title: Giardia and Cryptosporidium in mammalian wildlife – current status and future needs

text: Environmental pollution with human and domestic-animal fecal material is recognized as a potential pathogen pathway for wildlife infections with zooanthropomorphic protozoan parasites such as Giardia and Cryptosporidium. In this article, we review current knowledge about the diversity of free-living and captive terrestrial and marine mammalian wildlife species infected with Giardia and Cryptosporidium. The combination of prevalence studies with modern molecular-genotyping techniques is providing valuable insights into the host specificity and possible transmission routes of these two important parasites.


Clinical Characteristics and Outcomes of COVID-19 Among Patients with Pre-Existing Liver Disease in United States: A Multi-Center Research Network Study


title: Mechanisms and Pathophysiological Roles of the ATG8 Conjugation Machinery

text: Since their initial discovery around two decades ago, the yeast autophagy-related (Atg)8 protein and its mammalian homologues of the light chain 3 (LC3) and γ-aminobutyric acid receptor associated proteins (GABARAP) families have been key for the tremendous expansion of our knowledge about autophagy, a process in which cytoplasmic material become targeted for lysosomal degradation. These proteins are ubiquitin-like proteins that become directly conjugated to a lipid in the autophagy membrane upon induction of autophagy, thus providing a marker of the pathway, allowing studies of autophagosome biogenesis and maturation. Moreover, the ATG8 proteins function to recruit components of the core autophagy machinery as well as cargo for selective degradation. Importantly, comprehensive structural and biochemical in vitro studies of the machinery required for ATG8 protein lipidation, as well as their genetic manipulation in various model organisms, have provided novel insight into the molecular mechanisms and pathophysiological roles of the mATG8 proteins. Recently, it has become evident that the ATG8 proteins and their conjugation machinery are also involved in intracellular pathways and processes not related to autophagy. This review focuses on the molecular functions of ATG8 proteins and their conjugation machinery in autophagy and other pathways, as well as their links to disease.


title: Why obesity, hypertension, diabetes, and ethnicities are common risk factors for COVID‐19 and H1N1 influenza infections

text: Obesity, hypertension, diabetes, and specific ethnicities (Black and Hispanic) have been reported to be common comorbidities and possible risk factors for the severity of both COVID-19 and H1N1 influenza infections. Thus, it is important to understand why these four risk factors are common to both COVID-19 and H1N1 influenza infections, and whether a common mechanism exists. Respiratory failure is the most important pathology that contributes to the severity of both COVID-19 and H1N1 influenza infections. Additionally, obesity has been reported to be a risk factor for the development of acute respiratory distress syndrome (ARDS), which is a serious clinical manifestation of both COVID-19 and H1N1 infections. Obesity is a risk factor for hypertension. Most of the previous studies showing hypertension as a risk factor for the severity of COVID-19 and H1N1 infections were either not based on multiple logistic regression analyses or did not include obesity or BMI as an explanatory variable in their multiple logistic regression models. Moreover, similar attention is needed when specifying patients with diabetes or of specific ethnicities (Black and Hispanic) as potentially more vulnerable to either infection, because obesity also correlates with diabetes, and is more prevalent in these ethnicities. Notably, a retrospective cohort study has shown that obesity or high BMI are predictive risk factors for severe COVID-19 outcomes, independent of age, diabetes, and hypertension. Associations between hypertension, diabetes, ethnicities and severity of COVID-19 and H1N1 infections may be confounded by obesity to a considerable extent. This article is protected by copyright. All rights reserved.


title: Development of an indirect ELISA for detecting porcine deltacoronavirus IgA antibodies

text: Porcine deltacoronavirus (PDCoV) is a novel coronavirus that can cause vomiting and watery diarrhea in pigs and death in piglets. Since PDCoV was first detected in 2009 in Hong Kong, the prevalence of PDCoV has increased in recent years, resulting in serious economic losses to the swine industry. The coronavirus spike (S) protein is an antigen that has been demonstrated to contain epitopes that induce neutralizing antibodies. The presence of serum and milk IgA antibodies against pathogens that replicate primarily on mucosal surfaces is important for mucosal immunity. Here, an indirect anti-PDCoV IgA antibody enzyme-linked immunosorbent assay (PDCoV S1 IgA ELISA) using the purified S1 portion of S protein as the coating antigen was developed to detect PDCoV IgA antibodies in serum and sow’s milk. A receiver operating characteristic (ROC) curve analysis showed high specificity and sensitivity of the PDCoV-S1-IgA-ELISA based on samples confirmed by IFA. Anti-PDCoV IgA antibodies in 152 serum samples and 65 milk samples collected from six farms that had experienced diarrhea outbreaks within previous last two years were detected by this assay, and 62.5% of the serum samples and 100% of the milk samples were positive for PDCoV. The indirect ELISA method established in this study will provide a convenient tool for measurement of serum and milk IgA levels against PDCoV in pig herds, rapid detection of PDCoV infection in pigs, and evaluation of the immunogenicity of vaccines. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00705-020-04541-6) contains supplementary material, which is available to authorized users.


title: Repositioning PARP inhibitors for SARS‐CoV‐2 infection (COVID‐19); a new multi‐pronged therapy for ARDS?

text: Clinically approved PARP inhibitors (PARPi) have a mild adverse effect profile and are well‐tolerated as continuous daily oral therapy. We review the evidence that justifies the repurposing of PARPi to block the proliferation of SARS‐CoV‐2 and combat the life‐threatening sequelae of COVID‐19 by several mechanisms. PARPi’s can effectively decrease IL‐6, IL‐1 and TNFα levels (key interleukins in SARS‐CoV‐2‐induced cytokine storm) and can alleviate subsequent lung fibrosis, as demonstrated in murine experiments and clinical trials. PARPi can tune macrophages towards a tolerogenic phenotype. PARPi’s may also counteract SARS‐CoV‐2‐induced and inflammation‐induced cell death and support cell survival. PARPi’s had beneficial effects in animal models of acute respiratory distress syndrome (ARDS), asthma and ventilator‐induced lung injury. PARPi’s may potentiate the effectiveness of Tocilizumab, Anakinra, Sarilumab, Adalimumab, Canakinumab or Siltuximab therapy. In summary, the evidence suggests that PARPi therapy would benefit COVID‐19 patients and trials of these drugs should be undertaken.


title: In Silico Perspectives on the Prediction of the PLP’s Epitopes involved in Multiple Sclerosis

text: BACKGROUND: Multiple sclerosis (MS) is the most common autoimmune disease of the central nervous system (CNS). The main cause of the MS is yet to be revealed, but the most probable theory is based on the molecular mimicry that concludes some infections in the activation of T cells against brain auto-antigens that initiate the disease cascade. OBJECTIVES: The Purpose of this research is the prediction of the auto-antigen potency of the myelin proteolipid protein (PLP) in multiple sclerosis. MATERIALS AND METHODS: As there wasn’t any tertiary structure of PLP available in the Protein Data Bank (PDB) and in order to characterize the structural properties of the protein, we modeled this protein using prediction servers. Meta prediction method, as a new perspective in silico, was performed to fi nd PLPs epitopes. For this purpose, several T cell epitope prediction web servers were used to predict PLPs epitopes against Human Leukocyte Antigens (HLA). The overlap regions, as were predicted by most web servers were selected as immunogenic epitopes and were subjected to the BLASTP against microorganisms. RESULTS: Three common regions, AA(58-74), AA(161-177), and AA(238-254) were detected as immunodominant regions through meta-prediction. Investigating peptides with more than 50% similarity to that of candidate epitope AA(58-74) in bacteria showed a similar peptide in bacteria (mainly consistent with that of clostridium and mycobacterium) and spike protein of Alphacoronavirus 1, Canine coronavirus, and Feline coronavirus. These results suggest that cross reaction of the immune system to PLP may have originated from a bacteria or viral infection, and therefore molecular mimicry might have an important role in the progression of MS. CONCLUSIONS: Through reliable and accurate prediction of the consensus epitopes, it is not necessary to synthesize all PLP fragments and examine their immunogenicity experimentally (in vitro). In this study, the best encephalitogenic antigens were predicted based on bioinformatics tools that may provide reliable results for researches in a shorter time and at a lower cost.


title: Nitric oxide and peroxynitrite have different antiviral effects against hantavirus replication and free mature virions

text: Reactive nitrogen intermediates (RNI), like nitric oxide (NO) and peroxynitrite, have antiviral effects against certain viruses. Hantaviruses, like other members of the Bunyaviridae family, have previously not been shown to be sensitive to RNI. In this study, we compared the effects of NO and peroxynitrite on hantavirus replication and free mature virions in vitro, and of inducible nitric oxide synthase (iNOS) in hantavirus‐infected suckling mice. The NO‐generating compound S‐nitroso‐N‐acetylpenicillamine (SNAP), as well as cytokine‐induced NO, strongly inhibited hantavirus replication in Vero E6 cells, while pretreatment of free virions with SNAP only had a limited effect on their viability. In contrast, 3‐morpholinosydnonimine hydrochloride (SIN‐1), a peroxynitrite donor, inhibited virus replication only to a very low extent in vitro, but pretreatment of virus with SIN‐1 led to a considerably lowered viability of the virions. Infections of various human cell types per se did not induce NO production. The viral titers in iNOS(–/–) mice were higher compared to the controls, suggesting that NO inhibits hantavirus replication in vivo. In conclusion, we show that NO has strong antiviral effects on hantavirus replication, and peryoxynitrite on mature free virions, suggesting that different RNI can have different effects on various parts of the replication cycle for the same virus.


title: Tocilizumab for treatment of mechanically ventilated patients with COVID-19

text: BACKGROUND: Severe COVID-19 can manifest in rapid decompensation and respiratory failure with elevated inflammatory markers. This presentation is consistent with cytokine release syndrome in chimeric antigen receptor T cell therapy, for which IL-6 blockade is approved treatment. METHODS: We assessed effectiveness and safety of IL-6 blockade with tocilizumab in a single-center cohort of patients with COVID-19 requiring mechanical ventilation. The primary endpoint was survival probability post-intubation; secondary analyses included an ordinal illness severity scale integrating superinfections. Outcomes in patients who received tocilizumab compared to tocilizumab-untreated controls were evaluated using multivariable Cox regression with propensity score inverse probability weighting (IPTW). FINDINGS: 154 patients were included, of whom 78 received tocilizumab and 76 did not. Median follow-up was 47 days (range 28-67). Baseline characteristics were similar between groups, although tocilizumab-treated patients were younger (mean 55 vs. 60 years), less likely to have chronic pulmonary disease (10% vs. 28%), and had lower D-dimer values at time of intubation (median 2.4 vs. 6.5 mg/dL). In IPTW-adjusted models, tocilizumab was associated with a 45% reduction in hazard of death [hazard ratio 0.55 (95% CI 0.33, 0.90)] and improved status on the ordinal outcome scale [odds ratio per 1-level increase: 0.59 (0.36, 0.95)]. Though tocilizumab was associated with an increased proportion of patients with superinfections (54% vs. 26%; p<0.001), there was no difference in 28-day case fatality rate among tocilizumab-treated patients with versus without superinfection [22% vs. 15%; p=0.42]. INTERPRETATION: In this cohort of mechanically ventilated COVID-19 patients, tocilizumab was associated with a decreased likelihood of death despite higher superinfection occurrence. Randomized controlled trials are urgently needed to confirm these findings.


title: Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein

text: Abstract S1D (residues 636–789) is a neutralizing epitope region on the spike protein (S) of porcine epidemic diarrhea virus (PEDV). To accurately identify epitopes on S1D, the S1-phage library containing the gene encoding the S1D region of PEDV S protein was micropanned by six specific monoclonal antibodies (McAbs) against the S1D region. These micropanned epitope regions (MER) were focused on 696–779 amino acids of the S protein. To further map epitopes of the MER, seven overlapping mini-fragments covering MER nucleotides were separately synthesized and expressed in Escherichia coli BL21 with a GST tag. These mini-GST fusion proteins were scanned by ELISA and Western blotting with the six McAbs, and the result showed that S1D5 (residues 744–759) and S1D6 (residues 756–771) are two linear epitopes of the PEDV S protein. The antisera of the epitopes S1D5 and S1D6 could react with the native S protein of PEDV. Furthermore, Pepscan of the two linear epitopes demonstrated that SS2 (748YSNIGVCK755) and SS6 (764LQDGQVKI771) are two core epitopes on S1D5 and S1D6, respectively, located on the S protein of PEDV.


title: Microbial transmission in an outpatient clinic and impact of an intervention with an ethanol-based disinfectant

text: BACKGROUND: Halting the spread of harmful microbes requires an understanding of their transmission via hands and fomites. Previous studies explored acute and long-term care environments but not outpatient clinics. Objectives of this study were to track microbial movement throughout an outpatient clinic and evaluate the impact of a disinfectant spray intervention targeting high-touch point surfaces. METHODS: At the start of the clinic day, a harmless viral tracer was placed onto 2 fomites: a patient room door handle and front desk pen. Patient care, cleaning, and hand hygiene practices continued as usual. Facility fomites (n = 19), staff hands (n = 4), and patient hands (n = 3-4) were sampled after 2, 3.5, and 6 hours. Tracer concentrations at baseline (before intervention) were evaluated 6 hours after seeding. For the intervention trials, high-touch surfaces were cleaned 4 hours after seeding with an ethanol-based disinfectant and sampled 2 hours after cleaning. RESULTS: At 2, 3.5, and 6 hours after seeding, virus was detected on all surfaces and hands sampled, with examination room door handles and nurses’ station chair arms yielding the highest concentrations. Virus concentrations decreased by 94.1% after the disinfectant spray intervention (P = .001). CONCLUSIONS: Microbes spread quickly in an outpatient clinic, reaching maximum contamination levels 2 hours after inoculation, with the highest contamination on examination room door handles and nurses’ station chairs. This study emphasizes the importance of targeted disinfection of high-touch surfaces.


title: Molecular pathogenesis of severe acute respiratory syndrome

text: Abstract The global outbreak in 2002–2003 of severe acute respiratory syndrome (SARS) posed a serious threat to public health and had a significant impact on socioeconomic stability. Although the global outbreak of SARS has been contained, there are serious concerns over its re-emergence and bioterrorism potential, and up to date, no specific treatment exists for this disease. Here we review the progress of studies on the pathogenesis of the disease, in particular, studies on the molecular level.


SARS Outbreak in Taiwan


title: The Influenza A Virus Endoribonuclease PA-X Usurps Host mRNA Processing Machinery to Limit Host Gene Expression

text: Many viruses shut off host gene expression to inhibit antiviral responses. Viral proteins and host proteins required for viral replication are typically spared in this process, but the mechanisms of target selectivity during host shutoff remain poorly understood. Using transcriptome-wide and targeted reporter experiments, we demonstrate that the influenza A virus endoribonuclease PA-X usurps RNA splicing to selectively target host RNAs for destruction. Proximity-labeling proteomics reveals that PA-X interacts with cellular RNA processing proteins, some of which are partially required for host shutoff. Thus, PA-X taps into host nuclear pre-mRNA processing mechanisms to destroy nascent mRNAs shortly after their synthesis. This mechanism sets PA-X apart from other viral host shutoff proteins that target actively translating mRNAs in the cytoplasm. Our study reveals a unique mechanism of host shutoff that helps us understand how influenza viruses suppress host gene expression.


title: Enzyme-linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine sera

text: Abstract An enzyme-linked immunosorbent assay (ELISA) for detecting serum antibodies to the porcine epidemic diarrhea coronavirus (PEDV) was established by using cell culture-grown PEDV as antigen for coating. Ultracentrifugation through 20 and 45% (w/w) sucrose cushion proved to be the best antigen purification method. Examination of 1024 swine sera showed a high specificity and a greater sensitivity of the ELISA, when compared with indirect immunofluorescence. Reference sera with high antibody titers to PEDV originated from two pigs experimentally infected with PEDV. Three different antigen purification methods and the advantages of the ELISA compared with an immunofluorescence test are discussed.


title: Respiratory Medicine in China Progress, Challenges, and Opportunities

text: The past century witnessed a rapid development of respiratory medicine in China. The major burden of respiratory disease has shifted from infectious diseases to chronic noninfectious diseases. Great achievements have been made in improving the national standard of clinical management of various respiratory diseases and in smoking control. The specialty of respiratory medicine is expanding into pulmonary and critical care medicine. Nevertheless, respiratory diseases remain a major public health problem, with new challenges such as air pollution and nosocomial infections. This review describes the history, accomplishments, new challenges, and opportunities in respiratory medicine in China.


title: Quarantine for SARS, Taiwan

text: During the 2003 outbreak of severe acute respiratory syndrome (SARS) in Taiwan, >150,000 persons were quarantined, 24 of whom were later found to have laboratory-confirmed SARS-coronavirus (SARS-CoV) infection. Since no evidence exists that SARS-CoV is infective before the onset of symptoms and the quarantined persons were exposed but not symptomatic, we thought the quarantine's effectiveness should be investigated. Using the Taiwan quarantine data, we found that the onset-to-diagnosis time of previously quarantined confirmed case-patients was significantly shortened compared to that for those who had not been quarantined. Thus, quarantine for SARS in Taiwan screened potentially infective persons for swift diagnosis and hospitalization after onset, thereby indirectly reducing infections. Full-scale quarantine measures implemented on April 28 led to a significant improvement in onset-to-diagnosis time of all SARS patients, regardless of previous quarantine status. We discuss the temporal effects of quarantine measures and other interventions on detection and isolation as well as the potential usefulness of quarantine in faster identification of persons with SARS and in improving isolation measures.


title: Glycyrrhizic‐Acid‐Based Carbon Dots with High Antiviral Activity by Multisite Inhibition Mechanisms

text: With the gradual usage of carbon dots (CDs) in the area of antiviral research, attempts have been stepped up to develop new antiviral CDs with high biocompatibility and antiviral effects. In this study, a kind of highly biocompatible CDs (Gly‐CDs) is synthesized from active ingredient (glycyrrhizic acid) of Chinese herbal medicine by a hydrothermal method. Using the porcine reproductive and respiratory syndrome virus (PRRSV) as a model, it is found that the Gly‐CDs inhibit PRRSV proliferation by up to 5 orders of viral titers. Detailed investigations reveal that Gly‐CDs can inhibit PRRSV invasion and replication, stimulate antiviral innate immune responses, and inhibit the accumulation of intracellular reactive oxygen species (ROS) caused by PRRSV infection. Proteomics analysis demonstrates that Gly‐CDs can stimulate cells to regulate the expression of some host restriction factors, including DDX53 and NOS3, which are directly related to PRRSV proliferation. Moreover, it is found that Gly‐CDs also remarkably suppress the propagation of other viruses, such as pseudorabies virus (PRV) and porcine epidemic diarrhea virus (PEDV), suggesting the broad antiviral activity of Gly‐CDs. The integrated results demonstrate that Gly‐CDs possess extraordinary antiviral activity with multisite inhibition mechanisms, providing a promising candidate for alternative therapy for PRRSV infection.


title: COVID-19 Disease: invasive ventilation

text: Abstract This article focuses on the critical care nurse’s role in the management of patients with COVID-19 who require invasive ventilation in order to improve outcomes and prevent complications. The nature of COVID-19 is such that many patients deteriorate rapidly and for members of this group requiring intubation and invasive ventilation, different approaches to airway management and ventilatory support are required. In order to reduce the risk of complications and an overview of invasive ventilation, including commonly used modes, potential complications, nursing care, weaning and extubation are all described. COVID-19 presents several challenges as the disease progresses, hypoxemia may worsen, and the patient can develop Acute Respiratory Distress Syndrome. Therefore, additional treatment strategies including the use of the prone position and the use of nitric oxide and prostacyclin nebulisers have been included. The strategies presented in this article are relevant to both critical care nurses and those re-deployed to intensive care units where nurses will inevitably be involved in the management of patients requiring invasive ventilation. Weaning these patients off invasive ventilation is multi-factorial and may be short or long term. A multi-disciplinary weaning plan, the principles, stages/phases, and speed of weaning with expected parameters prior extubation are explained. Planned and unplanned extubation with the serious complications of the latter as the patient may not be ready and may require emergency re-intubation resulting in setbacks should be avoided.


title: Antiviral Drugs for Viruses Other Than Human Immunodeficiency Virus

text: Most viral diseases, with the exception of those caused by human immunodeficiency virus, are self-limited illnesses that do not require specific antiviral therapy. The currently available antiviral drugs target 3 main groups of viruses: herpes, hepatitis, and influenza viruses. With the exception of the antisense molecule fomivirsen, all antiherpes drugs inhibit viral replication by serving as competitive substrates for viral DNA polymerase. Drugs for the treatment of influenza inhibit the ion channel M(2) protein or the enzyme neuraminidase. Combination therapy with Interferon-α and ribavirin remains the backbone treatment for chronic hepatitis C; the addition of serine protease inhibitors improves the treatment outcome of patients infected with hepatitis C virus genotype 1. Chronic hepatitis B can be treated with interferon or a combination of nucleos(t)ide analogues. Notably, almost all the nucleos(t) ide analogues for the treatment of chronic hepatitis B possess anti–human immunodeficiency virus properties, and they inhibit replication of hepatitis B virus by serving as competitive substrates for its DNA polymerase. Some antiviral drugs possess multiple potential clinical applications, such as ribavirin for the treatment of chronic hepatitis C and respiratory syncytial virus and cidofovir for the treatment of cytomegalovirus and other DNA viruses. Drug resistance is an emerging threat to the clinical utility of antiviral drugs. The major mechanisms for drug resistance are mutations in the viral DNA polymerase gene or in genes that encode for the viral kinases required for the activation of certain drugs such as acyclovir and ganciclovir. Widespread antiviral resistance has limited the clinical utility of M(2) inhibitors for the prevention and treatment of influenza infections. This article provides an overview of clinically available antiviral drugs for the primary care physician, with a special focus on pharmacology, clinical uses, and adverse effects.


title: Knowledge-based repositioning of the anti-HCV direct antiviral agent Sofosbuvir as SARS-CoV-2 treatment

text: The new human coronavirus named SARS-CoV-2 is a positive-sense RNA virus for which no specific drugs are currently available. A knowledge-based analysis strongly suggests a possible repositioning of the anti-HCV direct antiviral agent (DAA) Sofosbuvir as treatment for SARS-CoV-2. Indeed, the RNA-dependent RNA-polymerases (RdRp) of the two viruses show high sequence and structural homology, supporting the likelihood of binding the Sofosbuvir molecule with similar efficiency. Such a repositioning would allow the containment of the SARS-CoV-2 pandemic and limit the progression of disease to potentially deadly COVID19.


title: Socioeconomic, geographic and climatic risk factors for canine parvovirus infection and euthanasia in Australia

text: Abstract Infection of canids with canine parvovirus (CPV) can result in severe, often fatal disease. This study aimed to examine climatic, socioeconomic and geographic risk factors for CPV infection and CPV-associated euthanasia in Australia. Australian veterinary hospital responses (534; 23.5 %) to a national veterinary survey of CPV case occurrences and euthanasias in 2016 were used. Severe caseloads (>40 cases per annum) were reported by 26 (11 %) hospitals (median 60 cases; IQR 50–110). Case reporting, case numbers, and without-treatment euthanasia were significantly associated with disadvantage across all Socio-Economic Index for Areas quintiles (p < 0.0001) – the greater the disadvantage, the more reports. Strong negative correlations were found between case numbers and the Index of Relative Socioeconomic Disadvantage (rSP = –0.3357, p < 0.0001) and also between euthanasia and the Index of Education and Occupation (rSP = –0.3762, p < 0.0001). Hospitals in more remote areas were also more likely to report cases and to euthanize without treatment (p < 0.0001). Of the climate variables, temperature of the hottest month was most strongly positively correlated with case numbers (rSP = 0.421, p < 0.0001), and lower annual rainfall was associated with more case-reporting hospitals (p < 0.0001). These results confirm that socioeconomic disadvantage is a significant risk-factor for CPV infection and outcome, and high temperature may also contribute to risk.


title: RNA sensors of the innate immune system and their detection of pathogens

text: The innate immune system plays a critical role in pathogen recognition and initiation of protective immune response through the recognition of pathogen associated molecular patterns (PAMPs) by its pattern recognition receptors (PRRs). Nucleic acids including RNA and DNA have been recognized as very important PAMPs of pathogens especially for viruses. RNA are the major PAMPs of RNA viruses, to which most severe disease causing viruses belong thus posing a tougher challenge to human and animal health. Therefore, the understanding of the immune biology of RNA PRRs is critical for control of pathogen infections especially for RNA virus infections. RNA PRRs are comprised of TLR3, TLR7, TLR8, RIG‐I, MDA5, NLRP3, NOD2, and some other minorities. This review introduces these RNA PRRs by describing the cellular localizations, ligand recognitions, activation mechanisms, cell signaling pathways, and recognition of pathogens; the cross‐talks between various RNA PRRs are also reviewed. The deep insights of these RNA PRRs can be utilized to improve anti‐viral immune response. © 2017 IUBMB Life, 69(5):297–304, 2017


title: The First Case of Feline Infectious Peritonitis-like Pyogranuloma in a Ferret Infected by Coronavirus in Japan

text: A male ferret, which was purchased from abroad at 9 months of age, had shown significant weight loss starting at 13 months of age. The ferret subsequently showed decreasing motor activity and recumbency and was euthanized at 14 months of age. At necropsy, a white, quail egg-sized mass was found in the mesentery. Histopathologically, multifocal granulomas consisting of necrotic foci, macrophages, fibroblasts and plentiful fibrous connective tissues were observed in the mesenteric mass. Surrounding the granulomas, inflammatory cell infiltration consisting of neutrophils, lymphocytes and plasmacytes was observed diffusely and significantly. Immunohistochemistry revealed small numbers of macrophages around necrotic foci that were positively stained for anti-mouse feline coronavirus. Electron microscopically, the cytoplasm of the macrophages contained viral particles, which were identified as coronavirus. The histopathological features in this ferret were similar to those in cats with feline infectious peritonitis (FIP). This was the first case in ferrets in Japan.


title: Development of rhinovirus study model using organ culture of turbinate mucosa

text: Abstract To better understand the pathophysiology of rhinovirus (RV) infection, a development of a study model using organ culture of turbinate mucosa was sought. Inferior turbinate mucosal tissues were cultured using air–liquid interface methods, on a support of gelfoam soaked in culture media. RV-16 was applied to the mucosal surface and washed off, and histological changes were evaluated. The success of RV infection was assayed by semi-nested RT-PCR of the mucosal surface fluid taken 48h after incubation. Intracellular RVs were visualized by in situ hybridization (ISH). Secretion of the cytokines, IL-6 and IL-8, into the culture media was quantitated by ELISA. After 7 days of culture, the turbinate mucosae did not show significant damage. A PCR product indicating successful RV infection was detected in 5 out of 10 mucosal tissues. ISH showed a very small number of positively stained cells focally located in the epithelial layer. In the beginning 24h after infection, secretion of IL-6 and IL-8 into the culture media of infected mucosae was significantly greater than into the media of control mucosae. Our results indicate that the air–liquid interface organ culture of turbinate mucosa could serve as an acceptable in vitro model for studying RV infection.


title: Production of anti-Trichophyton rubrum egg yolk immunoglobulin and its therapeutic potential for treating dermatophytosis

text: Abstract The aim of this study was to estimate the therapeutic potential of specific egg yolk immunoglobulin (IgY) on dermatophytosis caused by Trichophyton rubrum. The IgY was produced by immunizing hens with cell wall proteins of T. rubrum, extracted from eggs by PEG precipitation and then purified by ammonium sulfate precipitation. The cross-reactivity (CR) with other fungi, growth inhibition on T. rubrum in vitro and therapeutic effect on T. rubrum infection in BALB/C mice of the specific IgY were then evaluated. Anti- T. rubrum cell wall proteins IgY (anti-trCWP IgY) presented a certain degree of cross-reactivity with different fungi. In the in vitro and in vivo activity researches, Anti-trCWP IgY showed a significant dose-dependent growth inhibitory effect on T. rubrum in vitro and a significant dose-dependent therapeutic effect on T. rubrum infection in BALB/C mice.


title: Dynamics of an epidemic model with spatial diffusion

text: Abstract Mathematical models are very useful in analyzing the spread and control of infectious diseases which can be used to predict the developing tendency of the infectious disease, determine the key factors and to seek the optimum strategies of disease control. As a result, we investigated the pattern dynamics of a spatial epidemic model with logistic growth. By using amplitude equation, we found that there were different types of stationary patterns including spotted, mixed, and stripe patterns, which mean that spatial motion of individuals can form high density of diseases. The obtained results can be extended in other related fields, such as vegetation patterns in ecosystems.


The nation‐state, class, digital divides and social anthropology


title: Preparing for the unseen threat of infectious disease

text: The spread of COVID‐19, the contagious respiratory disease that's brought the country to a halt, isn't the first, and won't be the last, time institutions of higher learning have to respond to the danger of rapidly spreading infection. Keeping students, faculty, and staff protected from this invisible threat is typically the job of an interdisciplinary team that includes campus public safety, health services, and, often, local agency partners.


title: Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins

text: Abstract A simple, cell-based, membrane fusion assay system that uses split green fluorescent proteins (spGFPs) as an indicator was developed. The attachment of the pleckstrin homology (PH) domain to the N-termini of each spGFP not only localized the reporter signal to the plasma membrane but also helped the stable expression of the smaller spGFP of seventeen amino acid residues. It was shown that this system allowed real-time monitoring of membrane fusion by HIV-1 envelope protein (Env) without the addition of external substrates. This method can be adapted to the analyses of other viral membrane fusion.


Convalescent Plasma for COVID‐19: back to the future


title: Chapter 95 Management of Infection in Patients With Kidney Transplant

text: Abstract With the pervasive nature of chronic kidney disease, kidney transplantation is likely to continue to increase in the coming years. There are many infectious risks related to kidney transplant, including reactivation of latent infections, surgical complications, infectious risks related to immunosuppression, and nosocomial and community-acquired infections. These are described classically via timeline with early infections (first month), middle (1 to 6 months), and late (after 6 months). Kidney transplant patients may suffer from infections secondary to a vast array of organisms, including bacteria, fungi, and viruses. Certain infections, particularly viral infections such as cytomegalovirus, Epstein-Barr virus, and BK virus, may portend acute and chronic implications of the infection and its subsequent impact on graft function. Critical care physicians and nephrologists caring for patients with a renal transplant must understand the broad array of possible infections, atypical presentations, and nuanced implications for appropriate evaluation and subsequent therapy, combined with the need for possible prophylaxis and/or suppression. Multidisciplinary teams, including transplant physicians and infectious diseases physicians, are encouraged strongly.


title: Influenza B, C and D Viruses

text: This article summarizes current knowledge on the related influenza B and C viruses and considers the few studies on the recently identified influenza D virus. We focus on the particular viral genome organizations, the viral propagation cycles, as well as structural and functional insight into the encoded viral gene products. This is complemented with comprehensive sections that address the evolutionary strategies and the epidemiological significance of these influenza virus types, as well as the current state of interventions available for their control.


title: Crystal structure of the SARS-CoV-2 non-structural protein 9, Nsp9

text: Summary Many of the SARS-CoV-2 proteins have related counterparts across the Severe Acute Respiratory Syndrome (SARS-CoV) family. One such protein is non-structural protein 9 (Nsp9), which is thought to mediate viral replication, overall virulence and viral genomic RNA reproduction. We sought to better characterise the SARS-CoV-2 Nsp9 and subsequently solved its X-ray crystal structure, in an apo-form and, unexpectedly, in a peptide-bound form with a sequence originating from a rhinoviral 3C protease sequence (LEVL). The SARS-CoV-2 Nsp9 structure revealed the high level of structural conservation within the Nsp9 family. The exogenous peptide binding site is close to the dimer interface and impacted the relative juxtapositioning of the monomers within the homodimer. We have established a protocol for the production of SARS-CoV-2 Nsp9, determined its structure and identified a peptide-binding site that warrants further study to understanding Nsp9 function.


title: Surveillance Sans Frontières: Internet-Based Emerging Infectious Disease Intelligence and the HealthMap Project

text: John Brownstein and colleagues discuss HealthMap, an automated real-time system that monitors and disseminates online information about emerging infectious diseases.


title: Private space, shared space and private housing prices in Hong Kong: An exploratory study

text: This exploratory study examines the relationship between internal space, shared space and private housing prices. Housing floor area is an ambiguous concept in Hong Kong because it covers a possible exaggeration of the amount of ‘private space’ exclusively enjoyed by the owner and an unidentifiable portion of ‘public space’ shared with other owners within the development. Using hedonic pricing models, this study has found that different distributions between private and shared space command different values from the housing buyers. Shared communal space generally exerts a downward pressure on housing prices. The buyers are willing to pay more for the private space and some desirable forms of communal space. A higher willingness-to-pay for the desirable attributes such as clubhouse indicates that the Hong Kong people are increasingly concerned about the quality of living space in the built environment. This study suggests a need of further research into the exact measurement and the different forms of housing space rather than simply taking the stated floor space figures for granted.


title: Enhanced induction of dendritic cell maturation and HLA-A*0201-restricted CEA-specific CD8(+) CTL response by exosomes derived from IL-18 gene-modified CEA-positive tumor cells

text: Dendritic cells (DC)-derived or tumor-derived exosomes are a population of nanometer sized membrane vesicles that can induce specific anti-tumor immunity. However, the immunogenic potential and efficiency of exosomes-based tumor vaccine are not satisfactory enough to achieve a curative effect in clinical trials. In this article we investigated whether IL-18 genetic modification of tumor cells can increase the efficacy of exosomes derived from IL-18 gene-modified tumor cells. We transfected carcinoembryonic antigen (CEA)-expressing tumor cells with a recombinant adenovirus encoding human IL-18 (AdhIL-18) and prepared the exosomes, Exo/IL-18, from IL-18 gene-modified tumor cells. We found that Exo/IL-18 naturally contain CEA and bioactive IL-18. Moreover, Exo-IL-18 are potent in chemoattracting DC and T cells, enhancing the proliferation and Th1 cytokine release of PBMC, and promoting the phenotypic and functional maturation of DC. Furthermore, Exo/IL-18-pulsed DC are quite potent to induce HLA-A*0201-restricted, CEA-specific CD8(+) CTL from the PBMC of HLA-A*0201 CEA(+) cancer patients in vitro. In almost all of these experiments, Exo/IL-18 show more potent functions than the conventionally prepared exosomes derived from parent tumor cells without IL-18 gene modification. Our findings suggest that Exo/IL-18 has more potent capability to induce specific anti-tumor immunity, and our strategy of IL-18 modification of exosomes is a feasible approach to develop exosomes-based tumor vaccines.


title: l-Arginine stimulates intestinal intraepithelial lymphocyte functions and immune response in chickens orally immunized with live intermediate plus strain of infectious bursal disease vaccine

text: Infectious bursal disease (IBD) continues to pose potential threat to poultry industry all over the world. The disease can spell disaster not only through its infection but also by break of immunity in chickens vaccinated for other diseases. l-Arginine, a ubiquitous, semi-essential amino acid has emerged as an imunostimulant from variety of human and animal studies. In the present study, we demonstrate the stimulatory effects of l-arginine on intestinal intraepithelial lymphocyte (iIELs) functions as well as on systemic immune response in chickens orally vaccinated with live intermediate plus (IP) strain of IBD vaccine. Challenge studies with virulent IBDV revealed complete (100%) protection in IP + l-arginine group compared with 80% protection recorded in IP strain vaccinated chickens. Functional activities of iIELs evaluated by cytotoxicity assay demonstrated significantly high percentage cytotoxicity in IP + l-arginine groups compared with IP group (P < 0.05). Proliferative response of iIELs against IBDV antigen and Con-A was also significantly higher in IP + l-arginine group. Similar results were obtained with peripheral blood mononuclear cell blastogenic response to IBDV and Con-A analyzed as an indicator of systemic cell-mediated immune response. Orally administered IP strain vaccine elicited good antibody titres in both the groups, IP and IP + l-arginine, however, the antibody titres were significantly higher in IP + l-arginine group compared with IP vaccinated group (P < 0.05). These results clearly demonstrate that l-arginine stimulates intestinal and systemic immune response against IBDV.


Presumptive asymptomatic COVID-19 carriers’ estimation and expected person-to-person spreading among repatriated passengers returning from China


title: Composite Monte Carlo decision making under high uncertainty of novel coronavirus epidemic using hybridized deep learning and fuzzy rule induction

text: Abstract In the advent of the novel coronavirus epidemic since December 2019, governments and authorities have been struggling to make critical decisions under high uncertainty at their best efforts. In computer science, this represents a typical problem of machine learning over incomplete or limited data in early epidemic Composite Monte-Carlo (CMC) simulation is a forecasting method which extrapolates available data which are broken down from multiple correlated/casual micro-data sources into many possible future outcomes by drawing random samples from some probability distributions. For instance, the overall trend and propagation of the infested cases in China are influenced by the temporal-spatial data of the nearby cities around the Wuhan city (where the virus is originated from), in terms of the population density, travel mobility, medical resources such as hospital beds and the timeliness of quarantine control in each city etc. Hence a CMC is reliable only up to the closeness of the underlying statistical distribution of a CMC, that is supposed to represent the behaviour of the future events, and the correctness of the composite data relationships. In this paper, a case study of using CMC that is enhanced by deep learning network and fuzzy rule induction for gaining better stochastic insights about the epidemic development is experimented. Instead of applying simplistic and uniform assumptions for a MC which is a common practice, a deep learning-based CMC is used in conjunction of fuzzy rule induction techniques. As a result, decision makers are benefited from a better fitted MC outputs complemented by min–max rules that foretell about the extreme ranges of future possibilities with respect to the epidemic.


title: The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy

text: Abstract Zika virus (ZIKV) is a human-pathogenic flavivirus that has recently emerged as a global public health threat. ZIKV infection may be associated with congenital malformations in infected fetuses and severe neurological and systemic complications in infected adults. There are currently limited treatment options for ZIKV infection. AR-12 (OSU-03012) is a celecoxib derivative cellular kinase inhibitor that has broad-spectrum antiviral activities. In this study, we investigated the antiviral activity and mechanism of AR-12 against ZIKV. We evaluated the in vitro anti-ZIKV activity of AR-12, using cell protection and virus yield reduction assays, in multiple clinically relevant cell lines, and the in vivo treatment effects of AR-12 in a lethal mouse model using type I interferon receptor-deficient A129 mice. AR-12 inhibited ZIKV strains belonging to both the African and Asian/American lineages in Huh-7 and/or neuronal cells. AR12's IC50 against ZIKV was consistently <2 μM in these cells. ZIKV-infected A129 mice treated with intraperitoneally or orally administered AR-12 had significantly higher survival rate (50.0%–83.3% vs 0%, P < 0.05), less body weight loss, and lower blood and tissue ZIKV RNA loads than untreated control A129 mice. These anti-ZIKV effects were likely the results of down-regulation of the PI3K/Akt pathway by AR-12. Clinical trials using the clinically available and broad-spectrum AR-12 as an empirical treatment should be considered especially for patients residing in or returning from areas endemic of ZIKV and other arboviral infections who present with an acute undifferentiated febrile illness.


title: Analysis of the clinical characteristics, drug treatments and prognoses of 136 patients with coronavirus disease 2019

text: WHAT IS KNOWN AND OBJECTIVE: Since the December 2019 discovery of several cases of coronavirus disease 2019 (COVID‐19) in Wuhan, China, the infection has spread worldwide. Our aim is to report on the clinical characteristics, treatments and prognoses of COVID‐19. METHODS: This was a retrospective, single‐centre, case series of 136 patients who were diagnosed with COVID‐19 at Wuhan Third Hospital in Wuhan, China, between 28 January 2020 and 12 February 2020. The clinical characteristics, laboratory tests, treatment features and prognoses were summarized. RESULTS AND DISCUSSION: The 136 patients were divided into a moderate (M) group (n = 103, 75.7%) and a severe and critical (SC) group (n = 33, 24.3%). There were significant differences in the incidences of concomitant chronic medical illnesses (eg, hypertension, diabetes and cardiovascular disease), fever, dry cough and dyspnoea among the two groups (P < .05). Compared with those in the M group, lymphocyte count (LYM) decreased significantly in the SC group, while the serum levels of C‐reactive protein (CRP), procalcitonin (PCT), creatinine (Cre), D‐dimer, lactic dehydrogenase (LDH), myoglobin (MB) and troponin I (cTnl) increased significantly in the SC group (P < .05). The main therapeutic drugs were antivirals, antibiotics, glucocorticoids, immunomodulators, traditional Chinese medicine preparations and symptomatic support drugs. There were significant differences in the incidences of shock, myocardial injury, acute respiratory distress syndrome (ARDS) and renal injury among the two groups (P < .05). Among the 136 patients, 99 (72.7%) were cured, 14 (10.3%) were transferred to other hospital and 23 (16.9%) died. WHAT IS NEW AND CONCLUSION: Elderly patients with chronic diseases are more likely to develop severe or critical COVID‐19 with multiple organ damage or systemic injuries. The improvement of LYM and CRP may be associated with the prognoses of COVID‐19. The combined use of three or more antiviral drugs is to be avoided. The combination of broad‐spectrum antibacterial drugs is not recommended and the risk of drug‐induced liver injury should be monitored. Throughout a patient's hospitalization, their treatment plan should be evaluated and adjusted according to their vital signs, clinical symptoms, laboratory tests and imaging changes. Patients should receive effective psychological counselling.


SARS-CoV-2 infection in conjunctival tissue – Authors' reply


title: Real-time Fluorescent PCR Techniques to Study Microbial–Host Interactions

text: Publisher Summary This chapter describes how real-time polymerase chain reaction (PCR) performs and how it may be used to detect microbial pathogens and the relationship they form with their host. Research and diagnostic microbiology laboratories contain a mix of traditional and leading-edge, in-house and commercial assays for the detection of microbes and the effects they impart upon target tissues, organs, and systems. The PCR has undergone significant change over the last decade, to the extent that only a small proportion of scientists have been able or willing to keep abreast of the latest offerings. The chapter reviews these changes. It discusses the second-generation of PCR technology—kinetic or real-time PCR, a tool gaining widespread acceptance in many scientific disciplines but especially in the microbiology laboratory.


title: Histopathology of Middle East respiratory syndrome coronovirus (MERS‐CoV) infection – clinicopathological and ultrastructural study

text: AIMS: The pathogenesis, viral localization and histopathological features of Middle East respiratory syndrome – coronavirus (MERS‐CoV) in humans are not described sufficiently. The aims of this study were to explore and define the spectrum of histological and ultrastructural pathological changes affecting various organs in a patient with MERS‐CoV infection and represent a base of MERS‐CoV histopathology. METHODS AND RESULTS: We analysed the post‐mortem histopathological findings and investigated localisation of viral particles in the pulmonary and extrapulmonary tissue by transmission electron microscopic examination in a 33‐year‐old male patient of T cell lymphoma, who acquired MERS‐CoV infection. Tissue needle biopsies were obtained from brain, heart, lung, liver, kidney and skeletal muscle. All samples were collected within 45 min from death to reduce tissue decomposition and artefact. Histopathological examination showed necrotising pneumonia, pulmonary diffuse alveolar damage, acute kidney injury, portal and lobular hepatitis and myositis with muscle atrophic changes. The brain and heart were histologically unremarkable. Ultrastructurally, viral particles were localised in the pneumocytes, pulmonary macrophages, renal proximal tubular epithelial cells and macrophages infiltrating the skeletal muscles. CONCLUSION: The results highlight the pulmonary and extrapulmonary pathological changes of MERS‐CoV infection and provide the first evidence of the viral presence in human renal tissue, which suggests tissue trophism for MERS‐CoV in kidney.


An alternative Continuous Positive Airway Pressure device for COVID-19 patients


title: Oligonucleotide therapy: An emerging focus area for drug delivery in chronic inflammatory respiratory diseases

text: Abstract Oligonucleotide-based therapies are advanced novel interventions used in the management of various respiratory diseases such as asthma and Chronic Obstructive Pulmonary Disease (COPD). These agents primarily act by gene silencing or RNA interference. Better methodologies and techniques are the need of the hour that can deliver these agents to tissues and cells in a target specific manner by which their maximum potential can be reached in the management of chronic inflammatory diseases. Nanoparticles play an important role in the target-specific delivery of drugs. In addition, oligonucleotides also are extensively used for gene transfer in the form of polymeric, liposomal and inorganic carrier materials. Therefore, the current review focuses on various novel dosage forms like nanoparticles, liposomes that can be used efficiently for the delivery of various oligonucleotides such as siRNA and miRNA. We also discuss the future perspectives and targets for oligonucleotides in the management of respiratory diseases.


title: Emerging perspectives in management and prevention of infections of the respiratory tract in infants and children

text: Abstract New products, new procedures, new information, and new legislation will have a significant impact on management and prevention of respiratory Infections in children. Current areas of investigation include the changing epidemiology (increased number of children In day care), concern about morbidity of common infections (hearing impairment and effect on development of speech and language due to otitis media), and new modes of microbiologic diagnosis (antigen detection). New antimicrobial agents have wider spectrums of activity, Increased concentrations in body fluids, and lesser toxicity than available drugs. New uses of old drugs are identified (value of erythromycin for Legionella pneumophila, Chlamydia trachomatis, and Mycoplasma pneumoniae). Increased usage of chemoprophylaxis for prevention of recurrences of acute otitis media follows publication of impressive results of recent studies. New conjugate polysaccharide vaccines are Immunogenic in young infants. Finally, and of major importance to children, physicians, and manufacturers, Is vaccine liability legislation, now in congressional committee.


title: High Risk of Influenza Virus Infection Among Swine Workers: Examining a Dynamic Cohort in China

text: BACKGROUND: China is thought to be a hotspot for zoonotic influenza virus emergence, yet there have been few prospective studies examining the occupational risks of such infections. METHODS: We present the first 2 years of data collected from a 5-year, prospective, cohort study of swine-exposed and -unexposed participants at 6 swine farms in China. We conducted serological and virological surveillance to examine evidence for swine influenza A virus infection in humans. RESULTS: Of the 658 participants (521 swine-exposed and 137 swine-unexposed), 207 (31.5%) seroconverted against at least 1 swine influenza virus subtype (swine H1N1 or H3N2). Swine-exposed participants’ microneutralization titers, especially those enrolled at confined animal feeding operations (CAFOs), were higher against the swine H1N1 virus than were other participants at 12 and 24 months. Despite elevated titers, among the 187 study subjects for whom we had complete follow-up, participants working at swine CAFOs had significantly greater odds of seroconverting against both the swine H1N1 (odds ratio [OR] 19.16, 95% confidence interval [CI] 3.55–358.65) and swine H3N2 (OR 2.97, 95% CI 1.16–8.01) viruses, compared to unexposed and non-CAFO swine workers with less intense swine exposure. CONCLUSIONS: While some of the observed increased risk against swine viruses may have been explained by exposure to human influenza strains, study data suggest that even with elevated preexisting antibodies, swine-exposed workers were at high risk of infection with enzootic swine influenza A viruses.