title: Characterization, pro-inflammatory response and cytotoxic profile of bioaerosols from urban and rural residential settings in Pune, India()

text: Microbiota associated with airborne particulate matter (PM) is an important indicator of indoor pollution as they can be pathogenic and cause serious health threats to the exposed occupants. Present study aimed to investigate the level of culturable microbes associated with PM and their toxicological characterization in urban and rural houses of Pune city. Highest concentration of bacterial aerosols observed to be associated with PM(10) size fraction in urban site (2136 ± 285 CFU/m(3)) whereas maximum fungal concentration has been measured in rural houses (1521 ± 302 CFU/m(3)). Predominantly found bacterial species were Bacillus sp., S. aureus, and Pseudomonas aeruginosa and fungal species were Aspergillus sp., Cladosporium sp., and Penicillium sp. in both urban and rural residential premises. Concentration of endotoxin measured using the kinetic Limulus Amebocyte Lysate assay exhibited that the level of endotoxin in both urban and rural sites are associated with household characteristics and the activities performed in indoor as well as outdoor. Cell free DTT assay confirmed the ability of these airborne microbes to induce the production of reactive oxygen species (ROS) varying along with the types of microorganisms. On exposure of A549 cells to airborne microbes, a significant decrease in cell viability was observed in terms of both necrosis and apoptosis pathway. Elevated production of nitric oxide (NO) and proinflammatory cytokines in epithelial cells and macrophages clearly suggest the inflammatory nature of these airborne microbes. Results derived from the present study demonstrated that the indoor air of urban and rural houses of Pune is contaminated in terms of microbial load. Therefore, attention should be paid to control the factors favoring the microbial growth in order to safeguard the health of exposed inhabitants.


title: Nasopharyngeal Swabs Are More Sensitive Than Oropharyngeal Swabs for COVID-19 Diagnosis and Monitoring the SARS-CoV-2 Load

text: Objective: Detection of SARS-CoV-2 by oropharyngeal swabs (OPS) and nasopharyngeal swabs (NPS) is an essential method for coronavirus disease 2019 (COVID-19) management. It is not clear how detection rate, sensitivity, and the risk of exposure for medical providers differ in two sampling methods. Methods: In this prospective study, 120 paired NPS and OPS specimens were collected from 120 inpatients with confirmed COVID-19. SARS-CoV-2 nucleic acid in swabs were detected by real-time RT-PCR. The SARS-CoV-2 detection rate, sensitivity, and viral load were analyzed with regards NPS and OPS. Sampling discomfort reported by patients was evaluated. Results: The SARS-CoV-2 detection rate was significantly higher for NPS [46.7% (56/120)] than OPS [10.0% (12/120)] (P < 0.001). The sensitivity of NPS was also significantly higher than that of OPS (P < 0.001). At the time of sampling, the time of detectable SARS-CoV-2 had a longer median duration (25.0 vs. 20.5 days, respectively) and a longer maximum duration (41 vs. 39 days, respectively) in NPS than OPS. The mean cycle threshold (Ct) value of NPS (37.8, 95% CI: 37.0–38.6) was significantly lower than that of OPS (39.4, 95% CI: 38.9–39.8) by 1.6 (95% CI 1.0–2.2, P < 0.001), indicating that the SARS-CoV-2 load was significantly higher in NPS specimens than OPS. Patient discomfort was low in both sampling methods. During NPS sampling, patients were significantly less likely to have nausea and vomit. Conclusions: NPS had significantly higher SARS-CoV-2 detection rate, sensitivity, and viral load than OPS. NPS could reduce droplets production during swabs. NPS should be recommended for diagnosing COVID-19 and monitoring SARS-CoV-2 load. Chinese Clinical Trial Registry, number: ChiCTR2000029883.


title: Considerations for development of pharmacy support models for COVID-19 alternate care sites

text: DISCLAIMER: In an effort to expedite the publication of articles related to the COVID-19 pandemic, AJHP is posting these manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. PURPOSE: Guidance on alternate care site planning based on the experience of a health-system pharmacy department in preparing for an expected surge in coronavirus disease 2019 (COVID-19) cases is provided. SUMMARY: In disaster response situations such as the COVID-19 pandemic, healthcare institutions may be compelled to transition to a contingency care model in which staffing and supply levels are no longer consistent with daily practice norms and, while usual patient care practices are maintained, establishment of alternate care sites (eg, a convention center) may be necessitated by high patient volumes. Available resources to assist hospitals and health systems in alternate care site planning include online guidance posted within the COVID-19 resources section of the US Army Corps of Engineers website, which provides recommended medication and supply lists; and the Federal Healthcare Resilience Task Force’s alternate care site toolkit, a comprehensive resource for all aspects of alternate care site planning, including pharmacy services. Important pharmacy planning issues include security and storage of drugs, state board of pharmacy and Drug Enforcement Administration licensing considerations, and staff credentialing, education, and training. Key medication management issues to be addressed in alternate site care planning include logistical challenges of supply chain maintenance, optimal workflow for compounded sterile preparations (eg, on-site preparation vs off-site preparation and delivery from a nearby hospital), and infusion pump availability and suitability to patient acuity levels. CONCLUSION: Planning for and operation of alternate care sites in disaster response situations should include involvement of pharmacists in key decision-making processes at the earliest planning stages.


title: Glucose and electrolyte supplementation of drinking water improve the immune responses of poults with inanition 1

text: Abstract Enteric disorders predispose poultry to malnutrition. The objectives of this paper were 1) to simulate the inanition of poult enteritis mortality syndrome by restricting feed intake and 2) to develop a drinking water supplement that supports the immune functions of poults with inanition. Poults were restricted to 14 g of feed/d for 7 d beginning at 14 d of age then fed ad libitum until 36 d (recovery). The control was fed ad libitum. During the feed-restriction period, duplicate groups of 6 poults received 1 of 5 drinking water treatments: 1) restricted feed, unsupplemented water; 2) restricted feed + electrolytes (RE); 3) RE + glucose + citric acid (REGC); 4) REGC + betaine (REGCB); or 5) REGCB + zinc-methionine (REGCBZ). Immunological functions were assessed by inoculating poults with SRBC and B. abortus (BA) antigen at 15, 22, and 29 d of age. Antibody (Ab) titers were determined 7 d later for primary, secondary, and recovery responses. The primary and secondary total Ab titers to SRBC for restricted feed were 4.71 and 6.16 log3, which where lower (P < 0.05) than for controls (8.00 and 9.66 log3) and the other treatments. The recovery Ab titer for controls was 10.7, significantly higher than restricted feed (8.71) and RE (8.10) groups but not different from other treatments. The primary total Ab responses to BA were significantly lower in the restricted feed and RE groups as compared with the control and other treatments. Although feed restriction of poults to maintenance reduces the humoral immune responses, these responses can be significantly improved by drinking water containing electrolytes and especially sources of energy such as glucose and citric acid.


title: Experience of Severe Acute Respiratory Syndrome in Singapore: Importation of Cases, and Defense Strategies at the Airport

text: BACKGROUND: The importation of SARS was responsible for the outbreaks in Singapore, Hong Kong, Vietnam and Canada at a time when this new disease had not been identified. We report the incidence and impact of cases of SARS imported to Singapore between 25 February and 31 May 2003, and describe national measures to prevent further importation. METHODS: Information on imported cases of SARS and measures taken at entry points to Singapore was retrieved from the Ministry of Health and the Civil Aviation Authority of Singapore. RESULTS: Of the 6 imported cases, which all occurred before screening measures were implemented at the airport, only the first resulted in extensive secondary transmission. Of 442,973 air passengers screened after measures were implemented, 136 were sent to a designated hospital for further SARS screening; none was diagnosed as having SARS. CONCLUSIONS: The SARS outbreak in Singapore can be traced to the first imported case. The absence of transmission from the other imported cases was probably a result of relatively prompt identification and isolation of cases, together with a low potential for transmission. New imported SARS cases therefore need not lead to major outbreaks if systems are in place to identify and isolate them early. Screening at entry points is costly, has a low yield and is not sufficient in itself, but may be justified in light of the major economic, social and international impact which even a single imported SARS case may have.


title: Effector CD8+ T cell-derived interleukin-10 enhances acute liver immunopathology

text: Background & Aims Besides secreting pro-inflammatory cytokines, chemokines and effector molecules, effector CD8+ T cells that arise upon acute infection with certain viruses have been shown to produce the regulatory cytokine interleukin (IL)-10 and, therefore, contain immunopathology. Whether the same occurs during acute hepatitis B virus (HBV) infection and role that IL-10 might play in liver disease is currently unknown. Methods Mouse models of acute HBV pathogenesis, as well as chimpanzees and patients acutely infected with HBV, were used to analyse the role of CD8+ T cell-derived IL-10 in liver immunopathology. Results Mouse HBV-specific effector CD8+ T cells produce significant amounts of IL-10 upon in vivo antigen encounter. This is corroborated by longitudinal data in a chimpanzee acutely infected with HBV, where serum IL-10 was readily detectable and correlated with intrahepatic CD8+ T cell infiltration and liver disease severity. Unexpectedly, mouse and human CD8+ T cell-derived IL-10 was found to act in an autocrine/paracrine fashion to enhance IL-2 responsiveness, thus preventing antigen-induced HBV-specific effector CD8+ T cell apoptosis. Accordingly, the use of mouse models of HBV pathogenesis revealed that the IL-10 produced by effector CD8+ T cells promoted their own intrahepatic survival and, thus supported, rather than suppressed liver immunopathology. Conclusion Effector CD8+ T cell-derived IL-10 enhances acute liver immunopathology. Altogether, these results extend our understanding of the cell- and tissue-specific role that IL-10 exerts in immune regulation. Lay summary: Interleukin-10 is mostly regarded as an immunosuppressive cytokine. We show here that HBV-specific CD8+ T cells produce IL-10 upon antigen recognition and that this cytokine enhances CD8+ T cell survival. As such, IL-10 paradoxically promotes rather than suppresses liver disease.


Fatal COVID‐19 in a Patient with End‐Stage Liver Disease Wait‐Listed for Liver Transplantation: An Evidence‐Based Review of COVID‐19 Screening Modalities Prior to Transplant


title: An Application of Harrison's System Theory Model to Spark a Rapid Telehealth Expansion in the time of COVID‐19

text: INTRODUCTION: In response to the COVID‐19 pandemic, health systems had to quickly adopt a process for enabling targeted and patient‐centered care delivery. This case study describes the utilization of Harrison's open‐systems model to create an approach for rapid adoption of existing telehealth technologies in a large scale academic medical center. METHODS: An internal group of organizational developers, was enlisted to enable this effort. Local networks were employed and organized into focus groups to rapidly assess and address barriers to adoption and informal interviews with executive leadership were conducted to align organizational goals. Interventions include rapid deployment of focused and data driven provider, staff and patient support bolstered by effective communication and resource management. RESULTS: There was an increase in the number of patient portal activation codes by 75% during the month of March. The number of activation codes generated expectedly decreased in April as many patients now had activated patient portals. The video visit volume as a result of provider self‐scheduling increased went from a baseline of 0 to over 600 clinical visits. DISCUSSION: Experienced organizational development programs can facilitate adoption of change. The faculty practice of CUIMC has years of experience with supporting wide scale operational change centered on technology. In this case, providing engaged networks with tailored content that is focused on the process and available technology promoted rapid adoption and optimization. CONCLUSION: In the setting of profound external pressure, experience with the ability to focus on tailoring training and support to the culture of the organization helped to rapidly increase the availability and success of telehealth visits for a large scale academic medical center.


Every Peddler Praises His Own Needle: Have Clinical Rules in the Diagnosis of Subarachnoid Hemorrhage Supplanted Lumbar Punctures Yet?: Answers to the July 2013 Journal Club Questions


title: Immunization with SARS-CoV S DNA vaccine generates memory CD4(+) and CD8(+) T cell immune responses

text: An effective vaccine for severe acute respiratory syndrome (SARS) will probably require the generation and maintenance of both humoral and cellular immune responses. It has been reported that after natural infection in humans and immunization in animals with SARS-CoV vaccine, antibody is produced and persistent for a long period of time. In the present study, mice were immunized i.m. with SARS-CoV S DNA vaccine, and three different methods (ELISA, ELISPOT and FACS) were used to evaluate the immune responses when the cells were stimulated in vitro with a pool of peptides overlapping entire SARS spike protein. The results show that prime-immunization with SARS-CoV S DNA vaccine can induce both CD4(+) and CD8(+) T cell responses. Boosting with the same vaccine enhances CD4(+) and CD8(+) T cell responses in both lymphoid and nonlymphoid organs and were persistent over two months. The SARS-CoV S-specific CD4(+) and CD8(+) T cells were CD62L(−), a marker for memory cells, and −30 to 50% of the cells expressed IL-7Rα (CD127), a marker for the capacity of effector cells to develop into memory cells. In addition, immunization with the DNA vaccine elicited high levels of antibody production. Taken together, these data demonstrate that immunization with SARS-CoV S DNA vaccine can generate antigen-specific humoral and cellular immune responses that may contribute to long-term protection.


Restructuring the surgical service during the COVID‐19 pandemic: experience from a tertiary institution in Singapore


title: Mannose 6–Phosphate Receptors Are Sorted from Immature Secretory Granules via Adaptor Protein AP-1, Clathrin, and Syntaxin 6–positive Vesicles

text: The occurrence of clathrin-coated buds on immature granules (IGs) of the regulated secretory pathway suggests that specific transmembrane proteins are sorted into these buds through interaction with cytosolic adaptor proteins. By quantitative immunoelectron microscopy of rat endocrine pancreatic β cells and exocrine parotid and pancreatic cells, we show for the first time that the mannose 6–phosphate receptors (MPRs) for lysosomal enzyme sorting colocalize with the AP-1 adaptor in clathrin-coated buds on IGs. Furthermore, the concentrations of both MPR and AP-1 decline by ∼90% as the granules mature. Concomitantly, in exocrine secretory cells lysosomal proenzymes enter and then are sorted out of IGs, just as was previously observed in β cells (Kuliawat, R., J. Klumperman, T. Ludwig, and P. Arvan. 1997. J. Cell Biol. 137:595–608). The exit of MPRs in AP-1/clathrin-coated buds is selective, indicated by the fact that the membrane protein phogrin is not removed from maturing granules. We have also made the first observation of a soluble N-ethylmaleimide–sensitive factor attachment protein receptor, syntaxin 6, which has been implicated in clathrin-coated vesicle trafficking from the TGN to endosomes (Bock, J.B., J. Klumperman, S. Davanger, and R.H. Scheller. 1997. Mol. Biol. Cell. 8:1261–1271) that enters and then exits the regulated secretory pathway during granule maturation. Thus, we hypothesize that during secretory granule maturation, MPR–ligand complexes and syntaxin 6 are removed from IGs by AP-1/clathrin-coated vesicles, and then delivered to endosomes.


title: ‘VIOLET’: a fluorescence-based simulation exercise for training healthcare workers in the use of personal protective equipment

text: Summary Background Healthcare workers caring for patients with high-consequence infectious diseases (HCIDs) require protection from pathogen exposure, for example by wearing personal protective equipment (PPE). Protection is acquired through the inherent safety of the PPE components, but also their safe and correct use, supported by adequate training and user familiarity. However, the evidence base for HCID PPE ensembles and any associated training is lacking, with subsequent variation between healthcare providers. Aim To develop an evidence-based assessment and training tool for evaluating PPE ensembles and doffing protocols, in the assessment of patients with suspected HCIDs. Methods VIOLET (Visualising Infection with Optimised Light for Education and Training) comprises a healthcare mannequin adapted to deliver simulated bodily fluids containing UV-fluorescent tracers. On demand and remotely operated, the mannequin projectile vomits (blue), coughs (red), has diarrhoea (yellow) and is covered in sweat (orange). Wearing PPE, healthcare staff participate in an HCID risk assessment and examination of the ‘patient’, thereby becoming exposed to these bodily fluids. Contamination of PPE is visualized and body-mapped under UV light before and after removal. Observational findings and participant feedback, around its use as a training exercise, is also recorded. Findings Significant contamination from different exposure events was seen, enabling evaluation of PPE and doffing procedures used. Observational data and participant feedback demonstrated its strengths and success as a training technique. Conclusion Simulation exercises using VIOLET provide evidence-based assessment of PPE ensembles, and are a valuable resource for training of healthcare staff in wearing and safe doffing of PPE.


title: Aerosolized Particle Reduction: A Novel Cadaveric Model and a Negative Airway Pressure Respirator (NAPR) System to Protect Health Care Workers From COVID-19

text: OBJECTIVES: This study aimed to identify escape of small-particle aerosols from a variety of masks using simulated breathing conditions. This study also aimed to evaluate the efficacy of a negative-pressure environment around the face in preventing the escape of small aerosolized particles. STUDY DESIGN: This study is an evaluation study with specific methodology described below. SETTING: This study was performed in our institution’s fresh tissue laboratory. SUBJECTS AND METHODS: A fixed cadaver head was placed in a controlled environment with a black background, and small-particle aerosols were created using joss incense sticks (mass-median aerosol diameter of 0.28 µ). Smoke was passed through the cadaver head, and images were taken with a high-resolution camera in a standardized manner. Digital image processing was used to calculate relative amounts of small-particle escape from a variety of masks, including a standard surgical mask, a modified Ambu mask, and our negative airway pressure respirator (NAPR). RESULTS: Significant amounts of aerosolized particles escaped during the trials with no mask, a standard surgical mask, and the NAPR without suction. When suction was applied to the NAPR, creating a negative-pressure system, no particle escape was noted. CONCLUSION: We present a new and effective method for the study of small-particle aerosols as a step toward better understanding the spread of these particles and the transmission of coronavirus disease 2019. We also present the concept of an NAPR to better protect health care workers from aerosols generated from the upper and lower airways.


Strategies for the management of gastrointestinal surgery in the COVID-19 pandemic


Immune Effects of Intracerebral Infection with Mouse Hepatitis Virus


title: Clinical impact of rapid molecular detection of respiratory pathogens in patients with acute respiratory infection

text: Abstract Background Acute respiratory infections (ARI) are a leading cause of morbidity and mortality worldwide. There is a need to demonstrate the clinical impact of using the new, rapid and sensitive molecular assays in prospectively designed studies. Objectives To study the impact on medical management of a rapid molecular assay in patients with respiratory infections. Study design A prospective, randomized, non-blinded study was performed in patients presenting to the Emergency Department during two respiratory seasons (2016–2017). Diagnosis was performed by FilmArray Respiratory Panel (FilmArray-RP) or by immunofluorescence assay (IFA). Results A total of 432 patients (156 children and 276 adults) were analyzed. Diagnosis with FilmArray-RP was associated with significant changes in medical management including withholding antibiotic prescriptions (OR:15.52, 95%CI:1.99–120.83 in adults and OR:12.23, 95%CI:1.56–96.09 in children), and reduction in complementary studies in children (OR:9.64, 95%CI:2.13–43.63) compared to IFA. Decrease in oseltamivir prescriptions was significantly higher in adults in the FilmArray-RP group (p = 0.042; OR:1.19, 95%CI:0.51-2.79) compared to adults managed with IFA. Diagnostic yield was significantly higher by FilmArray-RP (81%) than by IFA (31%)(p < 0.001). The median time from sample collection to reporting was 1 h 52 min by FilmArray-RP and 26 h by IFA (p < 0.001). Conclusions The high respiratory viruses’ detection rate and availability of results within two hours when using FilmArray-RP were associated with decreases in antibiotic prescriptions and complementary studies and more accurate use of oseltamivir.


title: The 2–5 A system: Modulation of viral and cellular processes through acceleration of RNA degradation

text: Abstract The 2–5A system is an RNA degradation pathway that can be induced by the interferons (IFNs). Treatment of cells with IFN activates genes encoding several double-stranded RNA (dsRNA)-dependent synthetases. These enzymes generate 5′-triphosphorylated, 2′,5′-phosphodiester-linked oligoadenylates (2–5A) from ATP. The effects of 2–5A in cells are transient since 2–5A is unstable in cells due to the activities of phosphodiesterase and phosphatase. 2–5A activates the endoribonuclease 2–5A-dependent RNase L, causing degradation of single-stranded RNA with moderate specificity. The human 2–5A-dependent RNase is an 83.5 kDa polypeptide that has little, if any, RNase activity, unless 2–5A is present. 2–5A binding to RNase L switches the enzyme from its off-state to its on-state. At least three 2′,5′-linked oligoadenylates and a single 5′-phosphoryl group are required for maximal activation of the RNase. Even though the constitutive presence of 2–5A-dependent RNase is observed in nearly all mammalian cell types, cellular amounts of 2–5A-dependent mRNA and activity can increase after IFN treatment. One well-established role of the 2–5A system is as a host defense against some types of viruses. Since virus infection of cells results in the production and secretion of IFNs, and since dsRNA is both a frequent product of virus infection and an activator of 2–5A synthesis, the replication of encephalomyocarditis virus, which produces dsRNA during its life cycle, is greatly suppressed in IFN-treated cells as a direct result of RNA decay by the activated 2–5A-clependent RNase. This review covers the organic chemistry, enzymology, and molecular biology of 2–5A and its associated enzymes. Additional possible biological roles of the 2–5A system, such as in cell growth and differentiation, human immunodeficiency virus replication, heat shock, atherosclerotic plaque, pathogenesis of Type I diabetes, and apoptosis, are presented.


Editorial: Why is modeling COVID-19 so difficult?


title: Characterisation of rubella virus hemagglutinin rosettes

text: Abstract Purified rubella virus treated with Triton X-100 was subjected to centrifugation in a sucrose density gradient containing nonionic detergent β-D-octylglucoside. The result of this treatment was the formation of hemagglutinating rosettes containing viral glycoproteins VP2 (50,000 mol. wt.) and VP3 (63,000 mol. wt.). The rosettes have a 26 S sedimentation coefficient and a density of 1.2 g/cm3 in sucrose. Electron microscopy revealed 15 nm rosettes with a hollow center. The molecular weight of the rosettes was extrapolated at 850,00


title: Managing the supportive care needs of those affected by COVID-19

text: Globally, the number of people affected by coronavirus disease 2019 (COVID-19) is rapidly increasing. In most (>80%), the illness is relatively mild and can be self-managed out of hospital. However, in about 20% the illness causes respiratory compromise severe enough to require hospital admission [1]. Patients with severe and critical disease need full active treatment. This may include oxygen for hypoxaemia and ventilatory support, along with optimal management of complications, e.g. super-imposed bacterial infection, and any underlying co-morbidities, e.g. chronic obstructive pulmonary disease, congestive heart failure. To date, no antiviral agent has shown to be effective in treating the disease [2].


title: Viral bronchiolitis

text: Summary Viral bronchiolitis is a common clinical syndrome affecting infants and young children. Concern about its associated morbidity and cost has led to a large body of research that has been summarised in systematic reviews and integrated into clinical practice guidelines in several countries. The evidence and guideline recommendations consistently support a clinical diagnosis with the limited role for diagnostic testing for children presenting with the typical clinical syndrome of viral upper respiratory infection progressing to the lower respiratory tract. Management is largely supportive, focusing on maintaining oxygenation and hydration of the patient. Evidence suggests no benefit from bronchodilator or corticosteroid use in infants with a first episode of bronchiolitis. Evidence for other treatments such as hypertonic saline is evolving but not clearly defined yet. For infants with severe disease, the insufficient available data suggest a role for high-flow nasal cannula and continuous positive airway pressure use in a monitored setting to prevent respiratory failure.


title: A serological assay to detect SARS-CoV-2 seroconversion in humans

text: SARS-Cov-2 (severe acute respiratory disease coronavirus 2), which causes Coronavirus Disease 2019 (COVID19) was first detected in China in late 2019 and has since then caused a global pandemic. While molecular assays to directly detect the viral genetic material are available for the diagnosis of acute infection, we currently lack serological assays suitable to specifically detect SARS-CoV-2 antibodies. Here we describe serological enzyme-linked immunosorbent assays (ELISA) that we developed using recombinant antigens derived from the spike protein of SARS-CoV-2. Using negative control samples representing pre-COVID 19 background immunity in the general adult population as well as samples from COVID19 patients, we demonstrate that these assays are sensitive and specific, allowing for screening and identification of COVID19 seroconverters using human plasma/serum as early as two days post COVID19 symptoms onset. Importantly, these assays do not require handling of infectious virus, can be adjusted to detect different antibody types and are amendable to scaling. Such serological assays are of critical importance to determine seroprevalence in a given population, define previous exposure and identify highly reactive human donors for the generation of convalescent serum as therapeutic. Sensitive and specific identification of coronavirus SARS-Cov-2 antibody titers may, in the future, also support screening of health care workers to identify those who are already immune and can be deployed to care for infected patients minimizing the risk of viral spread to colleagues and other patients.


title: Residue analysis of a CTL epitope of SARS-CoV spike protein by IFN-gamma production and bioinformatics prediction

text: BACKGROUND: Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by the novel coronavirus SARS-CoV. The T cell epitopes of the SARS CoV spike protein are well known, but no systematic evaluation of the functional and structural roles of each residue has been reported for these antigenic epitopes. Analysis of the functional importance of side-chains by mutational study may exaggerate the effect by imposing a structural disturbance or an unusual steric, electrostatic or hydrophobic interaction. RESULTS: We demonstrated that N50 could induce significant IFN-gamma response from SARS-CoV S DNA immunized mice splenocytes by the means of ELISA, ELISPOT and FACS. Moreover, S366-374 was predicted to be an optimal epitope by bioinformatics tools: ANN, SMM, ARB and BIMAS, and confirmed by IFN-gamma response induced by a series of S358-374-derived peptides. Furthermore, each of S366-374 was replaced by alanine (A), lysine (K) or aspartic acid (D), respectively. ANN was used to estimate the binding affinity of single S366-374 mutants to H-2 Kd. Y367 and L374 were predicated to possess the most important role in peptide binding. Additionally, these one residue mutated peptides were synthesized, and IFN-gamma production induced by G368, V369, A371, T372 and K373 mutated S366-374 were decreased obviously. CONCLUSIONS: We demonstrated that S366-374 is an optimal H-2 Kd CTL epitope in the SARS CoV S protein. Moreover, Y367, S370, and L374 are anchors in the epitope, while C366, G368, V369, A371, T372, and K373 may directly interact with TCR on the surface of CD8-T cells.


title: The infant airway microbiome in health and disease impacts later asthma development

text: The nasopharynx (NP) is a reservoir for microbes associated with acute respiratory illnesses (ARI). The development of asthma is initiated during infancy, driven by airway inflammation associated with infections. Here, we report viral and bacterial community profiling of NP aspirates across a birth cohort, capturing all lower respiratory illnesses during their first year. Most infants were initially colonized with Staphylococcus or Corynebacterium before stable colonization with Alloiococcus or Moraxella, with transient incursions of Streptococcus, Moraxella or Haemophilus marking virus-associated ARIs. Our data identify the NP microbiome as a determinant for infection spread to the lower airways, severity of accompanying inflammatory symptoms, and risk for future asthma development. Early asymptomatic colonization with Streptococcus was a strong asthma predictor, and antibiotic usage disrupted asymptomatic colonization patterns.


title: Sequential serological surveys in the early stages of the coronavirus disease epidemic: limitations and perspectives

text: INTRODUCTION: Estimates of the number of individuals infected by severe acute respiratory syndrome coronavirus 2 are important for health planning and establishment of expectations regarding herd immunity. METHODS: Seven testing rounds of a serological survey were conducted at 1-week intervals between April 19 and May 31, 2020 in Teresina municipality. RESULTS: Over the 7 weeks, serological positivity increased from 0.56% (95% confidence interval [CI]: 0.18%-1.30%) to 8.33% (95% CI: 6.61%-10.33%), representing 33-53 persons infected for each reported case. CONCLUSIONS: Serological screening may be an important tool for understanding the immunity of a population and planning community interventions.


title: A serological survey of common feline pathogens in free-living European wildcats (Felis silvestris) in central Spain

text: Twenty-five serum samples of 22 free-living European wildcats (Felis silvestris) captured from 1991 to 1993 in central Spain were tested for evidence of exposure to seven feline pathogens. All the wildcats but one (95.4%) presented evidence of contact with at least one of the agents (mean = 2.2). Contact with feline leukemia virus (FeLV) was detected in 81% of the wildcats (antibodies, 77%; antigen p27, 15%). Antibodies to feline calicivirus (FCV, 80%), feline herpesvirus (FHV, 20%), feline parvovirus (FPV, 18%), and Chlamydophila sp. (27%) were also detected. Analyses were negative for feline immunodeficiency virus and feline coronavirus. The probability of having antibodies to FPV was inversely related with the concentration of serum cholesterol and with a morphometric index of body condition. Similarity in the composition of antibodies against disease agents (number and identity of detected and undetected antibodies) was significantly higher in pairs of female wildcats than in pairs of males or heterosexual pairs, suggesting that females had a more homogeneous exposure to pathogens. Seroprevalence for FHV was higher in males than in females. Antibodies to FHV and Chlamydophila sp. were more frequent in winter than in other seasons. In addition, the mean similarity of the pathogen community between pairs of serum samples was higher if both wildcats were caught during the same season than if they were not. Mean similarity was lowest when serum samples obtained in winter were compared with those from spring or summer. The results suggest that some agents probably had a reservoir in domestic cats and may cause some undetected morbidity/mortality in the studied wildcat population, whereas others, such as FeLV and FCV, may be enzootic.


title: Alisporivir inhibits MERS- and SARS-coronavirus replication in cell culture, but not SARS-coronavirus infection in a mouse model

text: Abstract Currently, there is no registered treatment for infections with emerging zoonotic coronaviruses like SARS- and MERS-coronavirus. We here report that in cultured cells low-micromolar concentrations of alisporivir, a non-immunosuppressive cyclosporin A-analog, inhibit the replication of four different coronaviruses, including MERS- and SARS-coronavirus. Ribavirin was found to further potentiate the antiviral effect of alisporivir in these cell culture-based infection models, but this combination treatment was unable to improve the outcome of SARS-CoV infection in a mouse model. Nevertheless, our data provide a basis to further explore the potential of Cyp inhibitors as host-directed, broad-spectrum inhibitors of coronavirus replication.


title: MF59 formulated with CpG ODN as a potent adjuvant of recombinant HSP65-MUC1 for inducing anti-MUC1+ tumor immunity in mice

text: Abstract MF59 is an oil-in-water emulsion adjuvant approved for influenza vaccines for human use in Europe. Due to its Th2 inducing properties, MF59 is seldom tested for cancer vaccines. In this study, MF59 formulated with a C-type CpG oligodeoxynucleotide (YW002) was tested for its Th1 adjuvant activity to induce immune responses to HSP65-MUC1, a recombinant fusion protein incorporating a mycobacterial heat shock protein (HSP65) and mucin 1, cell surface associated (MUC1) derived peptide. Combination of YW002 with MF59 (MF59-YW002) could confer a potent Th1 biasing property to the adjuvant, which enhanced the immunogenicity of HSP65-MUC1 to induce significantly higher levels of specific IgG2c, increased IFN-γ mRNA expression in splenocytes and the generation of antigen-specific cytotoxic T lymphocytes in mice. When prophylactically applied, MF59-YW002 adjuvant containing HSP65-MUC1 inhibited the growth of MUC1+ B16 melanoma and prolonged the survival of tumor-bearing mice. In contrast, adjuvant containing MF59 with HSP65-MUC1 in the absence of YW002, promoted the growth of MUC1+ B16 melanoma in mice. These results suggest that MF59 plus CpG oligodeoxynucleotide might be developed as an efficient adjuvant for tumor vaccines against melanoma, and possibly other tumors.


title: Incidence of asymptomatic deep vein thrombosis in patients with COVID-19 pneumonia and elevated D-dimer levels()

text: AIM: An increased risk of venous thromboembolism (VTE) in patients with COVID-19 pneumonia admitted to intensive care unit (ICU) has been reported. Whether COVID-19 increases the risk of VTE in non-ICU wards remains unknown. We aimed to evaluate the burden of asymptomatic deep vein thrombosis (DVT) in COVID-19 patients with elevated D-dimer levels. METHOD: In this prospective study consecutive patients hospitalized in non-intensive care units with diagnosis of COVID-19 pneumonia and D-dimer > 1000 ng/mL were screened for asymptomatic DVT with complete compression doppler ultrasound (CCUS). The study was approved by the Institutional Ethics Committee. RESULTS: The study comprised 156 patients (65.4% male). All but three patients received standard doses of thromboprophylaxis. Median days of hospitalization until CCUS was 9 (IQR 5–17). CCUS was positive for DVT in 23 patients (14.7%), of whom only one was proximal DVT. Seven patients (4.5%) had bilateral distal DVT. Patients with DVT had higher median D-dimer levels: 4527 (IQR 1925-9144) ng/ml vs 2050 (IQR 1428-3235) ng/ml; p < 0.001. D-dimer levels > 1570 ng/mL were associated with asymptomatic DVT (OR 9.1; CI 95% 1.1–70.1). D-dimer showed an acceptable discriminative capacity (area under the ROC curve 0.72, 95% CI 0.61–0.84). CONCLUSION: In patients admitted with COVID-19 pneumonia and elevated D-dimer levels, the incidence of asymptomatic DVT is similar to that described in other series. Higher cut-off levels for D-dimer might be necessary for the diagnosis of DVT in COVID-19 patients.


title: Cardiovascular Implications of Fatal Outcomes of Patients With Coronavirus Disease 2019 (COVID-19)

text: IMPORTANCE: Increasing numbers of confirmed cases and mortality rates of coronavirus disease 2019 (COVID-19) are occurring in several countries and continents. Information regarding the impact of cardiovascular complication on fatal outcome is scarce. OBJECTIVE: To evaluate the association of underlying cardiovascular disease (CVD) and myocardial injury with fatal outcomes in patients with COVID-19. DESIGN, SETTING, AND PARTICIPANTS: This retrospective single-center case series analyzed patients with COVID-19 at the Seventh Hospital of Wuhan City, China, from January 23, 2020, to February 23, 2020. Analysis began February 25, 2020. MAIN OUTCOMES AND MEASURES: Demographic data, laboratory findings, comorbidities, and treatments were collected and analyzed in patients with and without elevation of troponin T (TnT) levels. RESULT: Among 187 patients with confirmed COVID-19, 144 patients (77%) were discharged and 43 patients (23%) died. The mean (SD) age was 58.50 (14.66) years. Overall, 66 (35.3%) had underlying CVD including hypertension, coronary heart disease, and cardiomyopathy, and 52 (27.8%) exhibited myocardial injury as indicated by elevated TnT levels. The mortality during hospitalization was 7.62% (8 of 105) for patients without underlying CVD and normal TnT levels, 13.33% (4 of 30) for those with underlying CVD and normal TnT levels, 37.50% (6 of 16) for those without underlying CVD but elevated TnT levels, and 69.44% (25 of 36) for those with underlying CVD and elevated TnTs. Patients with underlying CVD were more likely to exhibit elevation of TnT levels compared with the patients without CVD (36 [54.5%] vs 16 [13.2%]). Plasma TnT levels demonstrated a high and significantly positive linear correlation with plasma high-sensitivity C-reactive protein levels (β = 0.530, P < .001) and N-terminal pro–brain natriuretic peptide (NT-proBNP) levels (β = 0.613, P < .001). Plasma TnT and NT-proBNP levels during hospitalization (median [interquartile range (IQR)], 0.307 [0.094-0.600]; 1902.00 [728.35-8100.00]) and impending death (median [IQR], 0.141 [0.058-0.860]; 5375 [1179.50-25695.25]) increased significantly compared with admission values (median [IQR], 0.0355 [0.015-0.102]; 796.90 [401.93-1742.25]) in patients who died (P = .001; P < .001), while no significant dynamic changes of TnT (median [IQR], 0.010 [0.007-0.019]; 0.013 [0.007-0.022]; 0.011 [0.007-0.016]) and NT-proBNP (median [IQR], 352.20 [174.70-636.70]; 433.80 [155.80-1272.60]; 145.40 [63.4-526.50]) was observed in survivors (P = .96; P = .16). During hospitalization, patients with elevated TnT levels had more frequent malignant arrhythmias, and the use of glucocorticoid therapy (37 [71.2%] vs 69 [51.1%]) and mechanical ventilation (41 [59.6%] vs 14 [10.4%]) were higher compared with patients with normal TnT levels. The mortality rates of patients with and without use of angiotensin-converting enzyme inhibitors/angiotensin receptor blockers was 36.8% (7 of 19) and 25.6% (43 of 168). CONCLUSIONS AND RELEVANCE: Myocardial injury is significantly associated with fatal outcome of COVID-19, while the prognosis of patients with underlying CVD but without myocardial injury is relatively favorable. Myocardial injury is associated with cardiac dysfunction and arrhythmias. Inflammation may be a potential mechanism for myocardial injury. Aggressive treatment may be considered for patients at high risk of myocardial injury.


title: Resource impact of managing suspected Middle East respiratory syndrome patients in a UK teaching hospital

text: Summary Clinical challenges exist in the management of hospitalized patients returning to the UK with potential Middle East respiratory syndrome coronavirus (MERS-CoV) infection, particularly with its clinical overlap with influenza, as demonstrated in this case-series and cost-analysis review of returning Hajj pilgrims. These patients were hospitalized with acute febrile respiratory illness, initially managed as potential MERS-CoV infections, but were eventually diagnosed with influenza. Additional costs were small, yet enhanced infection prevention measures created significant burdens on isolation rooms and staff time. Planning for predictable events such as Hajj is important for resource management. Here, in-house MERS-CoV diagnostic testing would have facilitated earlier diagnosis and discharge.


title: Modeling of the Toll-like receptor 3 and a putative Toll-like receptor 3 antagonist encoded by the African swine fever virus

text: African swine fever virus (ASFV) is a large double-stranded DNA virus responsible for a lethal pig disease, to which no vaccine has ever been obtained. Its genome encodes a number of proteins involved in virus survival and transmission in its hosts, in particular proteins that inhibit signaling pathways in infected macrophages and, thus, interfere with the host's innate immune response. A recently identified novel ASFV viral protein (pI329L) was found to inhibit the Toll-like receptor 3 (TLR3) signaling pathway, TLR3 being a crucial “danger detector.” pI329L has been predicted to be a transmembrane protein containing extracellular putative leucine-rich repeats similar to TLR3, suggesting that pI329L might act as a TLR3 decoy. To explore this idea, we used comparative modeling and other structure prediction protocols to propose (a) a model for the TLR3–Toll-interleukin-1 receptor homodimer and (b) a structural fold for pI329L, detailed at atomistic level for its cytoplasmic domain. As this later domain shares only remote sequence relationships with the available TLR3 templates, a more complex modeling strategy was employed that combines the iterative implementation of (multi)threading/assembly/refinement (I-TASSER) structural prediction with expertise-guided posterior refinement. The final pI329L model presents a plausible fold, good structural quality, is consistent with the available experimental data, and it corroborates our hypothesis of pI329L being a TLR3 antagonist.


title: Health Care–Associated Infections and the Radiology Department

text: Abstract Health care–associated infections (HCAIs) are a significant concern for both health care workers (HCWs) and patients. They are a major contributing factor of disease in industrialized countries, and are responsible for significant morbidity, mortality, and a direct annual financial loss of $6-7 billion in North America alone. They are an increasingly challenging health issue due to multidrug-resistant pathogens such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci among others, along with an increasing number of susceptible patients. Over the last three decades, the risk of HCAIs has increased in the radiology department (RD) in part because of an increased number of patients visiting the department and an increase in the utilization of imaging modalities. In this review, we will discuss how patients and staff can be exposed to HCAIs in the RD, including contaminated inanimate surfaces, radiology equipment, and associated medical devices. As the role of medical imaging has extended from primarily diagnosis to include more interventions, the implementation and development of standardized infection minimization protocols and infection control procedures are vital in the RD, particularly in interventional radiology. With globalisation and the rapid movement of people regionally, nationally, and globally, there is greater risk of exposure to contagious diseases such as Ebola, especially if infected patients are undiagnosed when they travel. For effective infection control, advanced training and education of HCWs in the RD is essential. The purpose of this article is to provide an overview of HCAIs as related to activities of the RD. We will discuss the following major topics including the variety of HCAIs commonly encountered, the role of the RD in HCAIs, transmission of infections to patients and HCWs in the RD, standard infection prevention measures, and the management of susceptible/infected patients in the RD. We shall also examine the role of, and the preparedness of, HCWs, including RD technologists and interventional radiologists, who may be exposed to undiagnosed, yet infected patients. We shall conclude with a brief discussion of the role of further research related to HCAIs. Learning Objectives After the completion of this review article, the readers will • Understand the exposure and role of radiology department in health care–associated infections, • Know the causes/modes/transmission of infections in radiology department, • Be conscious of standard disinfection protocols, • Be aware of current and future strategies required for the effective control of health care–associated infection in the radiology department. This is a CME article and provides the equivalent of 2 hours of continuing education that may be applied to your professional development credit system. A 10-question multiple-choice quiz follows this reading. Please note that no formalized credit (category A) is available from CAMRT.


title: Operating Room Guide for Confirmed or Suspected COVID-19 Pregnant Patients Requiring Cesarean Delivery

text: We sought to provide a clinical practice protocol for our labor and delivery (L&D) unit, to care for confirmed or suspected COVID-19 patients requiring cesarean delivery. A multidisciplinary team approach guidance was designed to simplify and streamline the flow and care of patient with confirmed or suspected COVID-19 requiring cesarean delivery. A protocol was designed to improve staff readiness, minimize risks, and streamline care processes. This is a suggested protocol which may not be applicable to all health care settings but can be adapted to local resources and limitations of individual L&D units. Guidance and information are changing rapidly; therefore, we recommend continuing to update the protocol as needed. Key Points: Cesarean delivery for confirmed or suspected novel coronavirus disease 2019 (COVID-19) patients. Team-based approach for streamline care. Labor and delivery protocols for COVID-19 positive patients.


title: Computational modeling of the bat HKU4 coronavirus 3CL(pro) inhibitors as a tool for the development of antivirals against the emerging Middle East respiratory syndrome (MERS) coronavirus

text: The Middle East respiratory syndrome coronavirus (MERS‐CoV) is an emerging virus that poses a major challenge to clinical management. The 3C‐like protease (3CL(pro)) is essential for viral replication and thus represents a potential target for antiviral drug development. Presently, very few data are available on MERS‐CoV 3CL(pro) inhibition by small molecules. We conducted extensive exploration of the pharmacophoric space of a recently identified set of peptidomimetic inhibitors of the bat HKU4‐CoV 3CL(pro). HKU4‐CoV 3CL(pro) shares high sequence identity (81%) with the MERS‐CoV enzyme and thus represents a potential surrogate model for anti‐MERS drug discovery. We used 2 well‐established methods: Quantitative structure‐activity relationship (QSAR)‐guided modeling and docking‐based comparative intermolecular contacts analysis. The established pharmacophore models highlight structural features needed for ligand recognition and revealed important binding‐pocket regions involved in 3CL(pro)‐ligand interactions. The best models were used as 3D queries to screen the National Cancer Institute database for novel nonpeptidomimetic 3CL(pro) inhibitors. The identified hits were tested for HKU4‐CoV and MERS‐CoV 3CL(pro) inhibition. Two hits, which share the phenylsulfonamide fragment, showed moderate inhibitory activity against the MERS‐CoV 3CL(pro) and represent a potential starting point for the development of novel anti‐MERS agents. To the best of our knowledge, this is the first pharmacophore modeling study supported by in vitro validation on the MERS‐CoV 3CL(pro). HIGHLIGHTS: MERS‐CoV is an emerging virus that is closely related to the bat HKU4‐CoV. 3CL(pro) is a potential drug target for coronavirus infection. HKU4‐CoV 3CL(pro) is a useful surrogate model for the identification of MERS‐CoV 3CL(pro) enzyme inhibitors. dbCICA is a very robust modeling method for hit identification. The phenylsulfonamide scaffold represents a potential starting point for MERS coronavirus 3CL(pro) inhibitors development.


title: Severe SARS‐CoV‐2 pneumonia in a 58‐year‐old patient with HIV: a clinical case report from the Republic of Cyprus

text: HIV and severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) co‐infection is a major challenge for the clinicians as it urged the importance of developing an optimal pharmaceutical scheme and patient's management. The reports that have been recently published regarding the course of SARS‐CoV‐2 in patients with HIV are sparse. In this brief report we describe, our first single‐centre experience from a 58‐year‐old Caucasian male patient with HIV who developed a severe SARS‐CoV‐2 infection, including clinical characteristics, treatment, and outcomes. This article is protected by copyright. All rights reserved.


title: The cognitive consequences of the COVID-19 epidemic: collateral damage?

text: Recovery from coronavirus disease 2019 (COVID-19) will be principally defined in terms of remission from respiratory symptoms, however both clinical and animal studies have shown that coronaviruses may spread to the nervous system. A systematic search on previous viral epidemics revealed that while there has been relatively little research in this area, clinical studies have commonly reported neurological disorders and cognitive difficulties. Little is known with regard to their incidence, duration or underlying neural basis. The hippocampus appears to be particularly vulnerable to coronavirus infections, thus increasing the probability of post-infection memory impairment, and acceleration of neurodegenerative disorders such as Alzheimer’s disease. Future knowledge of the impact of COVID-19, from epidemiological studies and clinical practice, will be needed to develop future screening and treatment programmes to minimize the long-term cognitive consequences of COVID-19.


title: On the Emergence of Cryptococcus gattii in the Pacific Northwest: Ballast Tanks, Tsunamis, and Black Swans

text: The appearance of Cryptococcus gattii in the North American Pacific Northwest (PNW) in 1999 was an unexpected and is still an unexplained event. Recent phylogenomic analyses strongly suggest that this pathogenic fungus arrived in the PNW approximately 7 to 9 decades ago. In this paper, we theorize that the ancestors of the PNW C. gattii clones arrived in the area by shipborne transport, possibly in contaminated ballast, and established themselves in coastal waters early in the 20th century. In 1964, a tsunami flooded local coastal regions, transporting C. gattii to land. The occurrence of cryptococcosis in animals and humans 3 decades later suggests that adaptation to local environs took time, possibly requiring an increase in virulence and further dispersal. Tsunamis as a mechanism for the seeding of land with pathogenic waterborne microbes may have important implications for our understanding of how infectious diseases emerge in certain regions. This hypothesis suggests experimental work for its validation or refutation.


title: Oral immunization with LacVax® OmpA induces protective immune response against Shigella flexneri 2a ATCC 12022 in a murine model

text: Abstract Shigellosis is an acute invasive disease of the lower intestine, which afflicts millions of people worldwide with an estimated one million fatalities per annum. Despite of extensive research during the last two decades, a vaccine against multi-drug resistant Shigella is not yet available in the market. To provide a safe, effective and broad-spectrum vaccine against Shigella, we explored food grade bacteria Lactococcus lactis (L. lactis) for the delivery of conserved antigenic protein; Outer membrane protein A (OmpA) to the mucosal sites for effective elicitation of systemic and mucosal immunity. We have previously confirmed the immunogenic potential of recombinant L. lactis expressing OmpA (LacVax® OmpA) in BALB/c mice. In the present study, we have characterized the humoral and cellular immune profile of LacVax® OmpA and assessed its protective efficacy using a newly developed human like murine shigellosis model. The significant increase in OmpA specific serum IgG, fecal sIgA and a Th1 dominant immune response (indicated by high INF-γ/IL-4 ratio) in LacVax® OmpA immunized mice revealed successful activation of humoral and cellular immunity. The LacVax® OmpA immunized animals were also protected from human-like shigellosis when challenged with S. flexneri 2a ATCC 12022. The antigen specific serum IgG, fecal sIgA, INF-γ and IL-10 levels were found to be the significant correlates of protection. Collectively these results suggest that the LacVax® OmpA is a promising prophylactic candidate against shigellosis. However, the protective efficacy of LacVax® OmpA in the higher animals would further strengthen its future application in humans.


title: Vaginal Lactobacillusgasseri CMUL57 can inhibit herpes simplex type 2 but not Coxsackievirus B4E2

text: This study aimed at demonstrating the antiviral activity of Lactobacillus gasseri CMUL57 (L. gasseri CMUL57), L. acidophilus CMUL67 and L. plantarum CMUL140 against herpes simplex type 2 (HSV-2) and Coxsackievirus B4E2 (CVB4E2), which are enveloped and naked viruses, respectively. These lactobacilli were non-cytotoxic and were able to reduce the cytopathic effect induced by HSV-2 in Vero cell monolayers. However, lactobacilli were not active against CVB4E2. Tested lactobacilli displayed anti-HSV-2 activity when they were co-incubated with the virus prior to inoculating the mixture to Vero cell monolayers. The detection of HSV-2 DNA by PCR in pellets of bacteria/virus mixtures let us to hypothesize that anti-HSV-2 activity of lactobacilli resulted from the viruses’ entrapment. This study showed the capabilities of vaginal lactobacilli to inhibit enveloped viruses such as HSV-2.


title: Urocystitis and Ureteritis in Holstein Calves with Septicaemia Caused by Salmonella enterica Serotype Dublin

text: Summary Salmonellosis is an enteric or multisystemic disease of global distribution that affects numerous animal species. Although Salmonella enterica has been associated with urinary tract lesions in man, information on urocystitis/ureteritis in cattle caused by salmonellae is lacking. This communication describes lesions of the inferior urinary tract in four Holstein calves with septicaemia caused by S. enterica subsp. enterica serotype Dublin. Examination of the urinary bladder revealed either diffuse irregular thickening (three cases) or petechiation (one case) of the mucosa. On histopathological examination, urocystitis with submucosal histiocytic, lymphocytic and plasmacytic infiltration and neutrophil transmigration through the urothelium was noted in all cases. In one case, a fibrinosuppurative ureteritis was detected. Salmonella Dublin was identified by culture, 16S rDNA sequencing and serotyping and Salmonella antigen was detected intralesionally by immunohistochemistry. Other lesions, indicative of septicaemia included hepatitis, enteritis, pericarditis, splenitis, lymphadenitis and pneumonia. We conclude that S. Dublin can be uropathogenic in cattle with septicaemia.


title: Protecting prone positioned patients from facial pressure ulcers using prophylactic dressings: A timely biomechanical analysis in the context of the COVID‐19 pandemic

text: Prone positioning is used for surgical access and recently in exponentially growing numbers of coronavirus disease 2019 patients who are ventilated prone. To reduce their facial pressure ulcer risk, prophylactic dressings can be used; however, the biomechanical efficacy of this intervention has not been studied yet. We, therefore, evaluated facial soft tissue exposures to sustained mechanical loads in a prone position, with versus without multi‐layered silicone foam dressings applied as tissue protectors at the forehead and chin. We used an anatomically realistic validated finite element model of an adult male head to determine the contribution of the dressings to the alleviation of the sustained tissue loads. The application of the dressings considerably relieved the tissue exposures to loading. Specifically, with respect to the forehead, the application of a dressing resulted in 52% and 71% reductions in soft tissue exposures to effective stresses and strain energy densities, respectively. Likewise, a chin dressing lowered the soft tissue exposures to stresses and strain energy densities by 78% and 92%, respectively. While the surgical context is clear and there is a solid, relevant need for biomechanical information regarding prophylaxis for the prone positions, the projected consequences of the coronavirus pandemic make the present work more relevant than ever before.


Impact of Nutritional Status and Anemia on COVID-19- Is it a public health concern? Evidence from National Family Health Survey-4 (2015-2016), India


title: Full genome sequence analysis of a newly emerged QX-like infectious bronchitis virus from Sudan reveals distinct spots of recombination

text: Abstract Infectious bronchitis virus (IBV) infection continues to cause economically important diseases in poultry while different geno- and serotypes continue to circulate globally. Two infectious bronchitis viruses (IBV) were isolated from chickens with respiratory disease in Sudan. Sequence analysis of the hypervariable regions of the S1 gene revealed a close relation to the QX-like genotype which has not been detected in Sudan before. Whole genome analysis of IBV/Ck/Sudan/AR251–15/2014 isolate by next generation sequencing revealed a genome size of 27,646 nucleotides harbouring 13 open reading frames: 5′-1a-1b-S-3a-3b-E-M-4b-4c-5a-5b-N-6b-3′. Highest nucleotide sequence identity of 93% for the whole genome was found with the Chinese IBV strain Ck/CH/LHLJ/140906, the Italian IBV isolate ITA/90254/2005 and the 4/91 vaccine strain. Phylogenetic analysis of the S1 gene revealed that the IBV/Ck/Sudan/AR251–15/2014 isolate clustered together with viruses of the GI-19 lineage. Recombination analysis gave evidence for distinct patterns of origin of RNA in the Sudanese isolate in multiple genes. Several sites of recombination were scattered throughout the genome suggesting that the Sudan-QX-like strain emerged as a unique recombinant from multiple recombination events of parental viruses from 4/91, H120 and ITA/90254/2005 genotypes. The Sudanese QX-like isolate is plausibly genetically different from IBV strains previously reported in Africa and elsewhere.


title: Use of Molecular Epidemiology in Veterinary Practice

text: Molecular epidemiology is a relatively new branch of epidemiology that uses molecular biology methods to study health and disease in populations. This article gives an introduction to molecular epidemiologic terminology and methodology and its usefulness in large animal medicine and veterinary public health. Applications in source tracing and vaccine studies and insights into transmission dynamics, host specificity, and niche adaptation of infectious organisms are presented. Examples are drawn from a variety of diseases, organisms, and host species and range from the global level to the individual-animal level.


title: Immune-mediated diseases of the dog and cat III. Immune-mediated diseases of the integumentary, urogenital, endocrine and vascular systems

text: Abstract The first two articles in this series have covered the basis of the immune mediated diseases, systemic immune-mediated diseases and those that involve the haemolymphatic and musculoskeletal system. The purpose of this article is to cover the immune-mediated diseases that involve the integumentary, urogenital, endocrine and vascular systems.


title: Structural basis for the differential classification of HLA-A*6802 and HLA-A*6801 into the A2 and A3 supertypes

text: Abstract High polymorphism is one of the most important features of human leukocyte antigen (HLA) alleles, which were initially classified by serotyping but have recently been re-grouped into supertypes according to their peptide presentation properties. Two relatively prevalent HLA alleles HLA-A*6801 and HLA-A*6802, are classified into the same serotype HLA-A68. However, based on their distinct peptide-binding characteristics, HLA-A*6801 is grouped into A3 supertype, whereas HLA-A*6802 belongs to A2 supertype, similar to HLA-A*0201. Thusfar, the structural basis of the different supertype definitions of these serotyping-identical HLA alleles remains largely unknown. Herein, we determined the structures of HLA-A*6801 and HLA-A*6802 presenting three typical A3 and A2 supertype-restricted peptides, respectively. The binding capabilities of these peptides to HLA-A*6801, HLA-A*6802, and HLA-A*0201 were analyzed. These data indicate that the similar conformations of the residues within the F pocket contribute to close-related peptide binding features of HLA-A*6802 and HLA-A*0201. However, the overall structure and the peptide conformation of HLA-A*6802 are more similar to HLA-A*6801 rather than HLA-A*0201 which illuminates the similar serotype grouping of HLA-A*6802 and HLA-A*6801. Our findings are helpful for understanding the divergent peptide presentation and virus-specific CTL responses impacted by MHC micropolymorphisms and also elucidate the molecular basis of HLA supertype definitions.


title: Changing health care worker behavior in relation to respiratory disease transmission with a novel training approach that uses biosimulation

text: BACKGROUND: This pilot study was conducted to determine whether supplementing standard classroom training methods regarding respiratory disease transmission with a visual demonstration could improve the use of personal protective equipment among emergency department nurses. METHODS: Participants included 20 emergency department registered nurses randomized into 2 groups: control and intervention. The intervention group received supplemental training using the visual demonstration of respiratory particle dispersion. Both groups were then observed throughout their work shifts as they provided care during January-March 2005. RESULTS: Participants who received supplemental visual training correctly utilized personal protective equipment statistically more often than did participants who received only the standard classroom training. CONCLUSION: Supplementing the standard training methods with a visual demonstration can improve the use of personal protective equipment during care of patients exhibiting respiratory symptoms.


title: Don’t be the “Fifth Guy”: Risk, Responsibility, and the Rhetoric of Handwashing Campaigns

text: In recent years, outbreaks such as H1N1 have prompted heightened efforts to manage the risk of infection. These efforts often involve the endorsement of personal responsibility for infection risk, thus reinforcing an individualistic model of public health. Some scholars—for example, Peterson and Lupton (1996)—term this model the “new public health.” In this essay, I describe how the focus on personal responsibility for infection risk shapes the promotion of hand hygiene and other forms of illness etiquette. My analysis underscores the use of constitutive and stigmatizing rhetoric to depict individual bodies, rather than environments, as prime sources of infection. Common among workplaces, this rhetoric provides the impetus for encouraging individual behavior change as a hedge against infection risk. I argue, though, that the mandating of personal responsibility for infection risk galvanizes a culture of stigma and blame that may work against the aims of public health.


title: Detection of Respiratory Tract Pathogens with Molecular Biology Methods

text: This paper describes the use in routine diagnosis of virological kit, which was designed to identify the 15 most common respiratory viruses in clinical specimens of nasopharyngeal aspirates, swabs, and bronchoalveolar lavage. It is a one-step multiplex RT-PCR system for the detection of influenza virus type A and type B, human respiratory syncytial virus type A, B; human adenovirus, human metapneumovirus, human coronaviruses 229E/NL63 and OC43, human parainfluenza type 1, 2, 3, human rhinovirus type A, B, human enterovirus, and bocavirus 1, 2, 3, 4. The article presents research conducted on the basis of swabs collected from patients who came to the Ear, Nose, and Throat Emergency Care Unit at the Department of Otolaryngology, Military Medical Institute in Warsaw, in February 2013. Due to the nature of work in an laryngological emergency ward, the material was collected only from those patients who reported problems associated with rhinitis or any dysfunction of the upper respiratory tract. The study shows that patients who came to seek laryngological assistance were usually infected with viruses having affinity for the airway epithelium.


title: Impact of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection and disease -2019 (COVID-19) on the quality of life of rheumatoid arthritis patients in Benin

text: Abstract Aim of the work: To study the impact on the quality of life (QoL) of rheumatoid arthritis (RA) patients following up in a university hospital in Benin during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) disease 2019 (COVID-19) pandemic. Patients and methods The QoL of 68 RA patients was assessed using Medical Outcome Study Short Form 36 (MOS-SF36) scale across the physical (PCS) and mental (MCS) components scores. The disease activity score (DAS28) was considered. Patients accepting to participate and having a mean of communication via social network, in particular WhatsApp were included. Results The mean age of the patients was 49.9±12.1 years (11-83 years) and were 65 females and 3 males. The mean DAS-28 at the start of the restriction measures was 3.4±1.5, the mean PCS and MCS were 71.1±20.3 and 67.1±16.02 respectively and became 4.7±2.04, 38.1±4.96 and 36.8±3.8 respectively. After 2 months, none of the RA patients presented with classic symptoms of COVID-19 infection. 8 patients were screened by Rapid Diagnostic Test and Reverse Transcription‐Polymerase Chain Reaction. Only 1 case was positive but asymptomatic. All patients had altered QoL according to MOS-SF36. Factors associated with this deterioration were stress related to SARS-CoV2 isolation (p=0.001), stress of having an activity flare-up (p=0.001), fear of being stigmatized (p=0.009). The economic factors were signficantly associated with temporary unemployment, decrease in monthly income, and drug discontinuation (p=0.001, p=0.002 and p=0.046 respectively). Conclusion The impact of the SARS-CoV2 pandemic is negative on the QoL of RA patients and many factors were contributing.


title: Homegrown Ultraviolet Germicidal Irradiation for Hospital-Based N95 Decontamination during the COVID-19 Pandemic

text: Coronavirus disease (COVID-19), the disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, is responsible for the 2020 global pandemic and characterized by high transmissibility and morbidity. Healthcare workers (HCWs) are at risk of contracting COVID-19, and this risk is mitigated through the use of personal protective equipment such as N95 Filtering Facepiece Respirators (FFRs). The high demand for FFRs is not currently met by global supply chains, potentially placing HCWs at increased exposure risk. Effective FFR decontamination modalities exist, which could maintain respiratory protection for HCWs in the midst of the current pandemic, through the decontamination and re-use of FFRs. Here, we present a locally-implemented ultraviolet-C germicidal irradiation (UVGI)-based FFR decontamination pathway, utilizing a home-built UVGI array assembled entirely with previously existing components available at our institution. We provide recommendations on the construction of similar systems, as well as guidance and strategies towards successful institutional implementation of FFR decontamination.


title: Microheterogeneity of S-glycoprotein of mouse hepatitis virus temperature-sensitive mutants

text: Abstract Mouse hepatitis virus (MHV) strain JHM (MHV-JHM) is a neurotropic coronavirus that causes acute fatal encephalomyelitis in 75–99% of infected mice. The surviving animals may subsequently develop demyelinating disease. We compared the S peplomer protein of the wild type (wt) and five temperature-sensitive (ts) mutants of MHV-JHM. In contrast with the wt, none of these five cause fatal disease (mortality less than 10%). Three of these ts mutants did not induce any demyelinating disease, a fourth caused demyelinating disease in 5% of the animals and a fifth, designated ts8, exhibited strong demyelinating properties and caused demyelination in 99% of the animals. SDS-PAGE analysis revealed no differences in the molecular weight of S peplomer protein of wt or ts MHV-JHM mutants. However, isoelectric focusing of the S protein of these five ts mutants and the wt MHV-JHM, followed by transfer to nitrocellulose sheets and immunoblotting with anti-S specific antibody revealed significant differences in the microheterogeneity of the S protein.


title: Clinical trial on type of calving pen and the risk of disease in Holstein calves during the first 90d of life

text: Abstract The objective of this study was to evaluate the efficacy of single cow calving pens that are cleaned between calvings vs. multiple cow calving pens for the prevention of calf diarrhea (scours), respiratory disease (pneumonia) and morbidity attributable to any cause. Every other pregnant cow or heifer was moved to either the single cow calving pen (treatment) or the multiple cow calving pen (control) within 48–72h prior to actual calving. The calves born in the single cow calving pens were assigned to the treatment group while the calves born in the multiple calving pens were assigned to the control group. Fecal materials, placental remains, and any other conspicuous dirt were removed from the single cow calving pens between each calving prior to the introduction of the next pregnant cow. The calves were then separated from their dams within 2h of birth. Multiple cow calving pens were managed as usual at the producers’ discretion. Upon birth, the calf managers monitored each enrolled calf for signs of diarrhea, pneumonia plus other morbidity up to 90d of age. The effects of single cow calving pens (vs. multiple cow calving pens) that are cleaned between calvings on the risk of neonatal calf diseases were evaluated using multivariable logistic regression models. Risk of diarrhea (OR=0.93, P =0.75), pneumonia (OR=1.23, P =0.64), and morbidity due to any cause (OR=0.93, P =0.74) were not significantly different between calves born in single cow vs. multiple cow calving pens. The current study found that, given the management situation evaluated, calves born in single cow calving pens were no different than calves born in multiple cow calving pens with respect to calf diseases risk. Long-term follow-up of the calves enrolled in the present study is ongoing to determine the efficacy of single cow calving pen use for the possible prevention of transmission of Mycobacterium avium subsp. paratuberculosis in Holstein calves.


title: First, second and potential third generation spreads of the COVID-19 epidemic in mainland China: an early exploratory study incorporating location-based service data of mobile devices

text: Abstract Objectives The outbreak of atypical pneumonia caused by the novel coronavirus (COVID-19) has currently become a global concern. The generations of the epidemic spread are not well known, yet these are critical parameters to facilitate an understanding of the epidemic. A seafood wholesale market and Wuhan city, China, were recognized as the primary and secondary epidemic sources. Human movements nationwide from the two epidemic sources revealed the characteristics of the first-generation and second-generation spreads of the COVID-19 epidemic, as well as the potential third-generation spread. Methods We used spatiotemporal data of COVID-19 cases in mainland China and two categories of location-based service (LBS) data of mobile devices from the primary and secondary epidemic sources to calculate Pearson correlation coefficient,r, and spatial stratified heterogeneity, q, statistics. Results Two categories of device trajectories had generally significant correlations and determinant powers of the epidemic spread. Bothr and q statistics decreased with distance from the epidemic sources and their associations changed with time. At the beginning of the epidemic, the mixed first-generation and second-generation spreads appeared in most cities with confirmed cases. They strongly interacted to enhance the epidemic in Hubei province and the trend was also significant in the provinces adjacent to Hubei. The third-generation spread started in Wuhan from January 17 to 20, 2020, and in Hubei from January 23 to 24. No obvious third-generation spread was detected outside Hubei. Conclusions The findings provide important foundations to quantify the effect of human movement on epidemic spread and inform ongoing control strategies. The spatiotemporal association between the epidemic spread and human movements from the primary and secondary epidemic sources indicates a transfer from second to third generations of the infection. Urgent control measures include preventing the potential third-generation spread in mainland China, eliminating it in Hubei, and reducing the interaction influence of first-generation and second-generation spreads.


title: Impact on mental health and perceptions of psychological care among medical and nursing staff in Wuhan during the 2019 novel coronavirus disease outbreak: A cross-sectional study

text: Abstract The severe 2019 outbreak of novel coronavirus disease (COVID-19), which was first reported in Wuhan, would be expected to impact the mental health of local medical and nursing staff and thus lead them to seek help. However, those outcomes have yet to be established using epidemiological data. To explore the mental health status of medical and nursing staff and the efficacy, or lack thereof, of critically connecting psychological needs to receiving psychological care, we conducted a quantitative study. This is the first paper on the mental health of medical and nursing staff in Wuhan. Notably, among 994 medical and nursing staff working in Wuhan, 36.9% had subthreshold mental health disturbances (mean PHQ-9: 2.4), 34.4% had mild disturbances (mean PHQ-9: 5.4), 22.4% had moderate disturbances (mean PHQ-9: 9.0), and 6.2% had severe disturbance (mean PHQ-9: 15.1) in the immediate wake of the viral epidemic. The noted burden fell particularly heavily on young women. Of all participants, 36.3% had accessed psychological materials (such as books on mental health), 50.4% had accessed psychological resources available through media (such as online push messages on mental health self-help coping methods), and 17.5% had participated in counseling or psychotherapy. Trends in levels of psychological distress and factors such as exposure to infected people and psychological assistance were identified. Although staff accessed limited mental healthcare services, distressed staff nonetheless saw these services as important resources to alleviate acute mental health disturbances and improve their physical health perceptions. These findings emphasize the importance of being prepared to support frontline workers through mental health interventions at times of widespread crisis.


OPTIMAL MANAGEMENT OF ELECTRODIAGNOSTIC STUDIES DURING COVID‐19 OUTBREAK


title: Antibody manufacture in transgenic animals and comparisons with other systems

text: Abstract Various forms of recombinant monoclonal antibodies are being used increasingly, mainly for therapeutic purposes. The isolation and engineering of the corresponding genes is becoming less of a bottleneck in the process; however, the production of recombinant antibodies is itself a limiting factor and a shortage is expected in the coming years. Milk from transgenic animals appears to be one of the most attractive sources of recombinant antibodies. None of the production systems presently implemented (CHO cells, insect cells infected by baculovirus, or transgenic animals and plants) has yet been optimized. This review describes the advantages of using milk for antibody production in comparison with the other systems.


title: Global Disease Detection—Achievements in Applied Public Health Research, Capacity Building, and Public Health Diplomacy, 2001–2016

text: The Centers for Disease Control and Prevention has established 10 Global Disease Detection (GDD) Program regional centers around the world that serve as centers of excellence for public health research on emerging and reemerging infectious diseases. The core activities of the GDD Program focus on applied public health research, surveillance, laboratory, public health informatics, and technical capacity building. During 2015–2016, program staff conducted 205 discrete projects on a range of topics, including acute respiratory illnesses, health systems strengthening, infectious diseases at the human–animal interface, and emerging infectious diseases. Projects incorporated multiple core activities, with technical capacity building being most prevalent. Collaborating with host countries to implement such projects promotes public health diplomacy. The GDD Program continues to work with countries to strengthen core capacities so that emerging diseases can be detected and stopped faster and closer to the source, thereby enhancing global health security.


Rehabilitation of Arthroplasty Patient During Covid-19


title: Early Use of Tocilizumab in Respiratory Failure Associated with Acute COVID -19 Pneumonia in Recipients with Solid Organ Transplantation

text: As of May 14, 2020, the World Health Organization has reported approximately 4.3 million cases of the novel Coronavirus Disease (COVID-19) with approximately 294,046 deaths worldwide.(1) Solid organ transplant recipients who are on chronic immunosuppressants fall within a special population of COVID-19 patients since they are more susceptible to complications secondary to COVID-19. Currently, we do not have data on treating COVID-19 patients with solid organ transplants with tocilizumab, an interleukin-6 (IL-6) inhibitor. We report a case of COVID-19 in a patient with a kidney and liver transplant and discuss the early use of tocilizumab to prevent the cytokine storm and attempt to reduce the likelihood of progression to Acute Respiratory Distress Syndrome (ARDS). In addition, we present other COVID-19 related transplant cases reported in the literature outlining the presenting clinical signs and outcomes.


A tentative guide for thoracic surgeons during COVID-19 pandemic


title: SARS Coronavirus Papain-Like Protease Inhibits the TLR7 Signaling Pathway through Removing Lys63-Linked Polyubiquitination of TRAF3 and TRAF6

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) papain-like protease (PLPro) reportedly inhibits the production of type I interferons (IFNs) and pro-inflammatory cytokines in Toll-like receptor 3 (TLR3) and retinoic acid-inducible gene 1 (RIG-I) pathways. The study investigated the inhibitory effect and its antagonistic mechanism of SARS-CoV PLPro on TLR7-mediated cytokine production. TLR7 agonist (imiquimod (IMQ)) concentration-dependently induced activation of ISRE-, NF-κB- and AP-1-luciferase reporters, as well as the production of IFN-α, IFN-β, TNF-α, IL-6 and IL-8 in human promonocyte cells. However, SARS-CoV PLPro significantly inhibited IMQ-induced cytokine production through suppressing the activation of transcription factors IRF-3, NF-κB and AP-1. Western blot analysis with anti-Lys48 and anti-Lys63 ubiquitin antibodies indicated the SARS-CoV PLPro removed Lys63-linked ubiquitin chains of TRAF3 and TRAF6, but not Lys48-linked ubiquitin chains in un-treated and treated cells. The decrease in the activated state of TRAF3 and TRAF6 correlated with the inactivation of TBK1 in response to IMQ by PLPro. The results revealed that the antagonism of SARS-CoV PLPro on TLR7-mediated innate immunity was associated with the negative regulation of TRAF3/6-TBK1-IRF3/NF-κB/AP1 signals.


title: Systemic Toxoplasmosis and Concurrent Porcine Circovirus-2 Infection in a Pig

text: Summary Systemic toxoplasmosis and concurrent infection with porcine circovirus-2 (PCV-2) was diagnosed in a fattening pig. Clinical examination of the herd showed that up to 30% of the pigs of this weight group suffered from severe respiratory signs including sneezing and coughing, with a mortality rate of up to 5%. Gross necropsy examination revealed severe interstitial pneumonia and generalized lymphadenopathy. On microscopical examination there was necrotizing inflammation of the lung, adrenal glands and lymph nodes, associated with lymphoid depletion, cytoplasmic basophilic botryoid inclusion bodies and protozoal microorganisms. Infection with Toxoplasma gondii was confirmed by immunohistochemistry (IHC). Polymerase chain reaction analysis, in-situ hybridization and IHC confirmed systemic PCV-2 infection. These findings, associated with the respiratory signs and lesions in lymphoid tissues, are characteristic for post-weaning multisystemic wasting syndrome (PMWS). In this case, immunosuppression by PCV-2 may have triggered systemic toxoplasmosis, or immune stimulation caused by coinfection with T. gondii may have caused extensive replication of PCV-2.


title: Porcine epidemic diarrhea virus (CV 777) and feline infectious peritonitis virus (FIPV) are antigenically related

text: Using gut sections from pigs infected with porcine epidemic diarrhea virus (strain CV 777) and ascitic fluid from cats which had succumbed to feline infectious peritonitis (FIP), a weak cross reaction was found by immunofluorescence. Its specificity was confirmed when detergent-treated purified CV 777 showed a prominent reaction with FIPV antibodies in ELISA; no reaction was obtained with intact virions, which indicated common determinants on an internal component of the particle. Antigenic cross-reactions at the nucleocapsid level were found in Western blot ELISA performed both ways (CV 777/FIPV antibodies; FIPV/CV 777 antibodies). In immunoprecipitation using [(35)S]methionine labelled FIPV, anti-CV 777 sera recognized exclusively the nucleocapsid protein. The significance of these findings for the classification of coronaviruses is discussed.


title: The role of adjuvant immunomodulatory agents for treatment of severe influenza

text: Abstract A severe inflammatory immune response with hypercytokinemia occurs in patients hospitalized with severe influenza, such as avian influenza A(H5N1), A(H7N9), and seasonal A(H1N1)pdm09 virus infections. The role of immunomodulatory therapy is unclear as there have been limited published data based on randomized controlled trials (RCTs). Passive immunotherapy such as convalescent plasma and hyperimmune globulin have some studies demonstrating benefit when administered as an adjunctive therapy for severe influenza. Triple combination of oseltamivir, clarithromycin, and naproxen for severe influenza has one study supporting its use, and confirmatory studies would be of great interest. Likewise, confirmatory studies of sirolimus without concomitant corticosteroid therapy should be explored as a research priority. Other agents with potential immunomodulating effects, including non-immune intravenous immunoglobulin, N-acetylcysteine, acute use of statins, macrolides, pamidronate, nitazoxanide, chloroquine, antiC5a antibody, interferons, human mesenchymal stromal cells, mycophenolic acid, peroxisome proliferator-activated receptors agonists, non-steroidal anti-inflammatory agents, mesalazine, herbal medicine, and the role of plasmapheresis and hemoperfusion as rescue therapy have supportive preclinical or observational clinical data, and deserve more investigation preferably by RCTs. Systemic corticosteroids administered in high dose may increase the risk of mortality and morbidity in patients with severe influenza and should not be used, while the clinical utility of low dose systemic corticosteroids requires further investigation.


title: Emerging trends in plasma-free manufacturing of recombinant protein therapeutics expressed in mammalian cells

text: Mammalian cells are the expression system of choice for therapeutic proteins, especially those requiring complex post-translational modifications. Traditionally, these cells are grown in medium supplemented with serum and other animal-or human-derived components to support viability and productivity. Such proteins are also typically added as excipients and stabilizers in the final drug formulation. However, the transmission of hepatitis B in the 1970s and of hepatitis C and HIV in the 1980s through plasma-derived factor VIII concentrates had catastrophic consequences for hemophilia patients. Thus, due to regulatory concerns about the inherent potential for transmission of infectious agents as well as the heterogeneity and lack of reliability of the serum supply, a trend has emerged to eliminate the use of plasma-derived additives in the production and formulation of recombinant protein therapeutics. This practice began with products used in the treatment of hemophilia and is progressively expanding throughout the entire industry. The plasma-free method of producing recombinant therapeutics is accomplished by the use of both cell culture media and final product formulations that do not contain animal-or human-derived additives. A number of recombinant therapeutic proteins for the treatment of several different diseases have been produced by plasma-free processes, with the objective of improving safety by eliminating blood-borne pathogens or by reducing immunogenicity. This review describes the factors that drove the development of plasma-free protein therapeutics and provides examples of advances in manufacturing that have made possible the removal of human and animal-derived products from all steps of recombinant protein production.


Enhancement of respiratory virus isolation from specimens using centrifugation and interferon inhibitors


DISNEA COMO SÍNTOMA DE ALARMA. IMPORTANCIA DE LA ATENCIÓN PRIMARIA EN UN NUEVO MODELO DE ATENCIÓN A PACIENTES COVID-19


title: Spondweni virus causes fetal harm in Ifnar1(-/-) mice and is transmitted by Aedes aegypti mosquitoes

text: Spondweni virus (SPONV) is the most closely related known flavivirus to Zika virus (ZIKV). Its pathogenic potential and vector specificity have not been well defined. SPONV has been found predominantly in Africa, but was recently detected in a pool of Culex quinquefasciatus mosquitoes in Haiti. Here we show that SPONV can cause significant fetal harm, including demise, comparable to ZIKV, in a mouse model of vertical transmission. Following maternal inoculation, we detected infectious SPONV in placentas and fetuses, along with significant fetal and placental histopathology, together suggesting vertical transmission. To test vector competence, we exposed Aedes aegypti and Culex quinquefasciatus mosquitoes to SPONV-infected bloodmeals. Aedes aegypti could efficiently transmit SPONV, whereas Culex quinquefasciatus could not. Our results suggest that SPONV has the same features that made ZIKV a public health risk.


title: Genomic characterization and infectivity of a novel SARS-like coronavirus in Chinese bats

text: SARS coronavirus (SARS-CoV), the causative agent of the large SARS outbreak in 2003, originated in bats. Many SARS-like coronaviruses (SL-CoVs) have been detected in bats, particularly those that reside in China, Europe, and Africa. To further understand the evolutionary relationship between SARS-CoV and its reservoirs, 334 bats were collected from Zhoushan city, Zhejiang province, China, between 2015 and 2017. PCR amplification of the conserved coronaviral protein RdRp detected coronaviruses in 26.65% of bats belonging to this region, and this number was influenced by seasonal changes. Full genomic analyses of the two new SL-CoVs from Zhoushan (ZXC21 and ZC45) showed that their genomes were 29,732 nucleotides (nt) and 29,802 nt in length, respectively, with 13 open reading frames (ORFs). These results revealed 81% shared nucleotide identity with human/civet SARS CoVs, which was more distant than that observed previously for bat SL-CoVs in China. Importantly, using pathogenic tests, we found that the virus can reproduce and cause disease in suckling rats, and further studies showed that the virus-like particles can be observed in the brains of suckling rats by electron microscopy. Thus, this study increased our understanding of the genetic diversity of the SL-CoVs carried by bats and also provided a new perspective to study the possibility of cross-species transmission of SL-CoVs using suckling rats as an animal model.


title: Small non-coding RNAs-based bone regulation and targeting therapeutic strategies

text: Abstract Small non-coding RNAs, which are 20–25 nucleotide ribonucleic acids, have emerged as an important transformation in the biological evolution over almost three decades. microRNAs (miRNAs) and short interfering RNAs (siRNAs) are two significant categories of the small RNAs that exert important effects on bone endocrinology and skeletology. Therefore, clarifying the expression and function of these important molecules in bone endocrine physiology and pathology is of great significance for improving their potential therapeutic value for metabolism-associated bone diseases. In the present review, we highlight the recent advances made in understanding the function and molecular mechanism of these small non-coding RNAs in bone metabolism, especially their potentially therapeutic values in bone-related diseases.


title: Antikörpertests bei COVID-19 - Was uns die Ergebnisse sagen

text: INTRODUCTION: In the context of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the detection of virus-specific antibodies (AB) will play an increasing role. The presence or absence of such antibodies can potentially lead to considerations regarding immunity and infection. ISSUE: How reliable are inferences from positive or negative test results regarding the actual presence of SARS-CoV-2 specific antibodies? METHODS: Calculation of the probability that, depending on the pretest probability (prevalence of SARS-CoV-2 infection) and test properties, antibodies are present or absent in the case of positive or negative test results. RESULTS: Sensitivity and specificity of different SARS-CoV-2 AB test systems vary between 53 % and 94 % and between 91 % and 99.5 %, respectively. When using a test with high test quality, the positive predictive value (PPV) is 42 % and 7 9%, respectively, with a pre-test probability of 1 % to 5 %, as can currently be assumed for the general population in Austria or Germany. For persons with an increased pre-test probability of 20 %, e. g. persons from high-risk professions, the PPW is 95 %, with a pre-test probability of 80 % the PPW is almost 100 %. The negative predictive value (NPV) is at least 99.7 % for persons with a low pre-test probability of up to 5 % and 79.1 % for persons with a pre-test probability of 80 %. When using test systems with lower sensitivity and specificity, the reliability of the results decreases considerably. The PPV is 5.9 % with a pre-test probability of 1 %. CONCLUSIONS: A sufficiently high sensitivity and specificity are prerequisites for the application of antibody test systems. Positive test results are often false if the pre-test probability is low. Depending on the assumed prevalence of a SARS-CoV-2 infection, there are substantial differences in the significance of a concrete test result for the respective affected persons.


Relation among right ventricular dysfunction, lung damage and mortality in patients with COVID-19


title: The Role Of Mitochondria In The Mammalian Antiviral Defense System

text: Innate immunity is a crucial defense system against viral and bacterial pathogens, providing a rapid response to mitigate the effects of microbial attack. While more readily associated with respiration and metabolism, recent research has surprisingly identified a number of mitochondrial factors in the mammalian innate immune system. This review summarizes the novel mitochondrial proteins, such as MAVS and NLRX1, involved in this process and attempts to reconcile this new mitochondrial function with our previous knowledge of the organelle.


title: Research Frontiers of Health Emergency and Disaster Risk Management: What Do We Know So Far?

text: Health-Emergency Disaster Risk Management (Health-EDRM) emerged as the latest knowledge, research and policy paradigm shift from response to preparedness and health risk management in non-emergency times [...].


title: Evolutionary Medicine: A Powerful Tool for Improving Human Health

text: Modern evolutionary research has much to contribute to medical research and health care practices. Conversely, evolutionary biologists are tapping into the rapidly expanding databases of medical genomic information to further their research. These two fields, which have historically functioned in almost complete isolation, are finding mutual benefit in the exchange of information. The long-term benefits of this synthesis of two major areas of research include improved health care. Recently, efforts to catalyze this relationship have brought together evolutionary biologists, medical practitioners, anthropologists, and ethicists to lay the groundwork for further collaboration and exploration. The range of overlap is surprisingly broad and potentially invaluable.


title: Simple and rapid detection of human enterovirus 71 by reverse-transcription and loop-mediated isothermal amplification: cryopreservation affected the detection ability

text: Abstract Human enterovirus 71 (EV71) is the primary pathogen of hand, foot, and mouth disease (HFMD). EV71 infection may lead to neurologic damage, with higher incidence of fatality compared with other HFMD pathogens. An effective drug or vaccine against EV71 infection is currently unavailable. It is desirable to determine the pathogen of HFMD accurately and quickly for early treatment. In the current study, reverse-transcription and loop-mediated isothermal amplification (RT-LAMP) technology were developed to detect EV71. The efficacy of detecting EV71 was compared with regular nested reverse-transcription polymerase chain reaction (RT-PCR). After detecting 108 clinical specimens, results showed that RT-LAMP can specifically detect EV71, but not Coxsackie virus A16, and exhibited a specificity of 100% and a sensitivity of 97.1%, which was higher than regular RT-PCR. The findings indicate that RT-LAMP is a practical method for EV71 diagnostic applications, particularly in small county institutes of medical service. The detection ability of RT-LAMP was significantly affected by cryopreservation as the clinical specimens were repeatedly subject to freezing and thawing treatments.


Leveraging Antisense Technology in the Lungs: RNA-Targeting Therapies for Respiratory Diseases