title: Potential Role of Anti-interleukin (IL)-6 Drugs in the Treatment of COVID-19: Rationale, Clinical Evidence and Risks

text: The epidemic due to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has been spreading globally, raising increasing concerns. This public health emergency has triggered a race to find medications to improve the prognosis of this disease. There is currently great interest in drug repositioning to manage SARS-CoV-2 infection, that is, the evaluation of the potential benefits of a drug that has already been proven safe and effective in humans for other approved indications. As interleukin-6 (IL-6) acts as a key driver of the inflammation associated with coronavirus disease 2019 (COVID-19), IL-6 and IL-6 receptor (IL-6R) inhibition appear to be promising targets for the treatment of COVID-19 patients. It is important to critically analyze the available evidence concerning the use of the available anti-IL-6 (siltuximab) and anti-IL-6R (tocilizumab and sarilumab) agents in COVID-19 patients, in terms of both benefit and risk. In this review, the pathogenesis of the cytokine storm induced by COVID-19, the role of IL-6 in this cytokine storm, the rationale for the use of anti-IL-6 agents, and key information on potential benefits and safety monitoring of these biologicals in COVID-19 patients is discussed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s40259-020-00430-1) contains supplementary material, which is available to authorized users.


title: The SARS-coronavirus nsp7+nsp8 complex is a unique multimeric RNA polymerase capable of both de novo initiation and primer extension

text: Uniquely among RNA viruses, replication of the ∼30-kb SARS-coronavirus genome is believed to involve two RNA-dependent RNA polymerase (RdRp) activities. The first is primer-dependent and associated with the 106-kDa non-structural protein 12 (nsp12), whereas the second is catalysed by the 22-kDa nsp8. This latter enzyme is capable of de novo initiation and has been proposed to operate as a primase. Interestingly, this protein has only been crystallized together with the 10-kDa nsp7, forming a hexadecameric, dsRNA-encircling ring structure [i.e. nsp(7+8), consisting of 8 copies of both nsps]. To better understand the implications of these structural characteristics for nsp8-driven RNA synthesis, we studied the prerequisites for the formation of the nsp(7+8) complex and its polymerase activity. We found that in particular the exposure of nsp8's natural N-terminal residue was paramount for both the protein's ability to associate with nsp7 and for boosting its RdRp activity. Moreover, this ‘improved’ recombinant nsp8 was capable of extending primed RNA templates, a property that had gone unnoticed thus far. The latter activity is, however, ∼20-fold weaker than that of the primer-dependent nsp12-RdRp at equal monomer concentrations. Finally, site-directed mutagenesis of conserved D/ExD/E motifs was employed to identify residues crucial for nsp(7+8) RdRp activity.


title: The outlook for diagnostic purposes of the 2019‐novel coronavirus disease

text: At the end of December 2019, a novel acute respiratory syndrome coronavirus 2 (SARS‐CoV2) appeared as the third unheard of outbreak of human coronavirus infection in the 21st century. First, in Wuhan, China, the novel SARS‐CoV2 was named by the World Health Organization (WHO), as 2019‐nCOV (COVID‐19), and spread extremely all over the world. SARS‐CoV2 is transmitted to individuals by human‐to‐human transmission leading to severe viral pneumonia and respiratory system injury. SARS‐CoV2 elicits infections from the common cold to severe conditions accompanied by lung injury, acute respiratory distress syndrome, and other organ destruction. There is a possibility of virus transmission from asymptomatic cases as active carriers, in addition to symptomatic ones, which is a crucial crisis of COVID‐19 that should be considered. Hence, paying more attention to the accurate and immediate diagnosis of suspected and infected cases can be a great help in preventing the rapid spread of the virus, improving the disease prognosis, and controlling the pandemic. In this review, we provide a comprehensive and up‐to‐date overview of the different types of Clinical and Para‐clinical diagnostic methods and their practical features, which can help understand better the applications and capacities of various diagnostic approaches for COVID‐19 infected cases.


title: Repositioning of 8565 Existing Drugs for COVID-19

text: [Image: see text] The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected over 7.1 million people and led to over 0.4 million deaths. Currently, there is no specific anti-SARS-CoV-2 medication. New drug discovery typically takes more than 10 years. Drug repositioning becomes one of the most feasible approaches for combating COVID-19. This work curates the largest available experimental data set for SARS-CoV-2 or SARS-CoV 3CL (main) protease inhibitors. On the basis of this data set, we develop validated machine learning models with relatively low root-mean-square error to screen 1553 FDA-approved drugs as well as another 7012 investigational or off-market drugs in DrugBank. We found that many existing drugs might be potentially potent to SARS-CoV-2. The druggability of many potent SARS-CoV-2 3CL protease inhibitors is analyzed. This work offers a foundation for further experimental studies of COVID-19 drug repositioning.


title: Development of a Luminex assay for the detection of swine antibodies to non-structural proteins of foot-and-mouth disease virus

text: Abstract Foot-and mouth disease (FMD), swine vesicular disease (SVD), and vesicular stomatitis (VS) are highly contagious vesicular diseases of swine but are not easy to differentiate clinically. For the purpose of instant detecting of FMD and differentiating it from the other vesicular diseases, a Luminex assay was developed. Sera from 64 infected, 307 vaccinated, and 280 naïve pigs were tested by the Luminex assay. Diagnostic sensitivity of the assay was 100%. Diagnostic specificity of the assay was 98.7% in vaccinated pigs and 97.5% to 100% in naïve pigs. Agreement between the results from the Luminex assay and those from a 3ABC polypeptide blocking ELISA was 96.3% with kappa statistics of 0.92. The Luminex assay can detect the immune response to NSP-3ABC in swine as early as eight days post-infection. Moreover, all of the 15 vaccinated but unprotected pigs were all detected by the Luminex assay. The results indicated that the Luminex assay has potential with specificity in detecting antibodies to FMDV 3ABC NSP and in distinguishing FMDV-infected pigs from with either SVDV or VSV.


title: Equine Arteritis Virus Subgenomic RNA Transcription: UV Inactivation and Translation Inhibition Studies

text: Abstract The expression of the genetic information of equine arteritis virus (EAV), an arterivirus, involves the synthesis of six subgenomic (sg) mRNAs. These are 5′ and 3′ coterminal since they are composed of a leader and a body sequence, which are identical to the 5′ and 3′ ends of the genome, respectively. Previously, it has been suggested thatcis-splicing of a genome-length precursor RNA is involved in their synthesis. This was reevaluated in a comparative analysis of the sg RNA synthesis of EAV, the coronavirus mouse hepatitis virus (MHV), and the alphavirus Sindbis virus. UV transcription mapping showed that the majority of the EAV sg RNAs made at later stages of infection is not derived from a genome-length precursor. However, complete independence of sg RNA synthesis from that of genomic RNA was never observed during the course of infection. The possibility that this resulted from UV irradiation-induced effects on the synthesis of the viral replicase was investigated by inhibiting translation using cycloheximide. For EAV, ongoing protein synthesis was found to be more important for the synthesis of sg RNA than for that of genomic RNA. In general, MHV transcription was extremely sensitive to translation inhibition, whereas EAV genomic RNA synthesis became independent ofde novoprotein synthesis late in infection.


title: Association between single nucleotide polymorphisms in TLR4, TLR2, TLR9, VDR, NOS2 and CCL5 genes with acute viral bronchiolitis

text: Abstract Background Acute viral bronchiolitis is the leading cause of hospitalization among infants during the first year of life. Most infants hospitalized for bronchiolitis do not present risk factors and are otherwise healthy. Our objective was to determine the genetic features associated with the risk and a severe course of bronchiolitis. Methods We prospectively evaluated 181 infants with severe bronchiolitis admitted at three hospitals over a 2-year period, who required oxygen therapy. The control group consisted of 536 healthy adults. Patients were evaluated for the presence of comorbidities (premature birth, chronic respiratory disease, and congenital heart disease), underwent nasopharyngeal aspirate testing for virus detection by multiplex-PCR, and SNPs identification in immune response genes. Patient outcomes were assessed. Results We observed association between SNP rs2107538*CCL5 and bronchiolitis caused by respiratory syncytial virus(RSV) and RSV-subtype-A, and between rs1060826*NOS2 and bronchiolitis caused by rhinovirus. SNPs rs4986790*TLR4, rs1898830*TLR2, and rs2228570*VDR were associated with progression to death. SNP rs7656411*TLR2 was associated with length of oxygen use; SNPs rs352162*TLR9, rs187084*TLR9, and rs2280788*CCL5 were associated with requirement for intensive care unit admission; while SNPs rs1927911*TLR4, rs352162*TLR9, and rs2107538*CCL5 were associated with the need for mechanical ventilation. Conclusions Our findings provide some evidence that SNPs in CCL5 and NOS2 are associated with presence of bronchiolitis and SNPs in TLR4, TLR2, TLR9, VDR and CCL5 are associated with severity of bronchiolitis.


title: Synthesis of distal and proximal fleximer base analogues and evaluation in the nucleocapsid protein of HIV-1

text: Abstract Anti-HIV-1 drug design has been notably challenging due to the virus’ ability to mutate and develop immunity against commercially available drugs. The aims of this project were to develop a series of fleximer base analogues that not only possess inherent flexibility that can remain active when faced with binding site mutations, but also target a non-canonical, highly conserved target: the nucleocapsid protein of HIV (NC). The compounds were predicted by computational studies not to function via zinc ejection, which would endow them with significant advantages over non-specific and thus toxic zinc-ejectors. The target fleximer bases were synthesized using palladium-catalyzed cross-coupling techniques and subsequently tested against NC and HIV-1. The results of those studies are described herein.


title: A rapid scoping review of Middle East respiratory syndrome coronavirus in animal hosts

text: Middle East respiratory syndrome coronavirus (MERS‐CoV) is an emerging zoonotic pathogen discovered in 2012. The purpose of this scoping review was to summarize the empirical evidence for MERS‐CoV in animals in order to map knowledge gaps and to extract data for modelling disease transmission in dromedary camels. A review protocol was developed a priori, and a systematic search, data extraction and summary were conducted using the Arksey and O'Malley framework. Ninety‐nine publications were identified for full review out of 1,368 unique records. Of these publications, 71 were articles in scientific journals. Ninety of the studies were observational and the remaining nine were experimental. We summarize characteristics of animal studies including study design, study population and outcomes of interest for future transmission modelling in the reservoir population. The majority of field studies reported measures of prevalence, while experimental studies provided estimates of transmission parameters that pertain to the natural course of disease.


title: Viral shedding dynamics in asymptomatic and mildly symptomatic patients infected with SARS-CoV-2

text: OBJECTIVES: Asymptomatic patients, together with those with mild symptoms of COVID-19, may play an important role in SARS-CoV-2 transmission. However, the dynamics of viral shedding during the various phases of the clinical course of COVID-19 remains unclear at this stage. METHODS: A total of 18 patients found positive for SARS-CoV-2 infection by real-time reverse transcription PCR (RT-PCR) assay, and admitted to Chongqing University Central Hospital between Jan 29 and Feb 5 were enrolled in this study. Medical data, pulmonary CT scan images, and RT-PCR results were periodically collected during the patients’ hospital stay. All participants were actively followed-up for 2 weeks after discharge. RESULTS: A total of nine (50%) asymptomatic patients and nine 9 (50%) mildly mild symptom COVID-19 patients were identified on admission. Six (66.7%) asymptomatic patients on admission developed subjective symptoms during hospitalization were recategorized as ‘pre-symptomatic’. The median duration of viral shedding was 11.5, 28 and 31 days for pre-symptomatic, asymptomatic and mild symptomatic patients, separately. Seven (38.9%) patients continued to shed virus after hospital discharge. During the convalescent phase, detectable antibodies to SARS-CoV-2 and RNA were simultaneously observed in five (27.8%) patients. CONCLUSIONS: Long-term viral shedding was presented in patients with mild symptoms and asymptomatic cases, and specific antibody production to SARS-CoV-2 may not guarantee viral clearance after discharge. These observations should be considered when making decisions regarding clinical and public health and strategies for prevention and control of SARS-CoV-2 infection.


Usefulness of lung ultrasound imaging in COVID‐19 pneumonia: The persisting need of safety and evidences


Intranasal application of polyethyleneimine suppresses influenza virus infection in mice


Pandemic … or syndemic? Re‐framing COVID‐19 disease burden and ‘underlying health conditions’


Dashboards to facilitate nephrology disaster planning in the COVID-19 era


title: Novel coronavirus (SARS‐CoV‐2) infection in a renal transplant recipient: Case report

text: An ongoing outbreak of pneumonia associated with the severe acute respiratory coronavirus 2 (SARS‐CoV‐2) started in Wuhan, China, with cases now confirmed in multiple countries. The clinical course of patients remains to be fully characterized, clinical presentation ranges from asymptomatic infection to acute respiratory distress syndrome and acute renal failure, and no pharmacological therapies of proven efficacy yet exist. We report a case of SARS‐CoV‐2 infection in a renal transplant recipient with excellent outcome. This case states the importance of close monitoring of the concentration of cyclosporine in patients treated with lopinavir/ritonavir; the routine treatment of corticosteroid can be continued. This is a rare report of SARS‐CoV‐2 infection in a renal transplant recipient. Further data are needed to achieve better understanding of the impact of immunosuppressive therapy on the clinical presentation, severity, and outcome of SARS‐CoV‐2 infections in solid organ transplant recipients.


title: Lower respiratory tract infections in cats Reaching beyond empirical therapy

text: Practical relevance Lower respiratory tract infections (LRTIs) in cats can be due to bacteria, parasites, fungi and viruses. This review details the practical investigation of these infections and highlights specific therapy where possible. The aim is to avoid the all-too-frequent temptation in practice to treat cats with lower respiratory tract signs empirically for feline bronchial disease (FBD)/asthma. This is potentially hazardous as immunosuppressive therapy for FBD/asthma could exacerbate disease due to a LRTI. Empirical treatment of suspected LRTI is also difficult to recommend given the wide range of potential pathogens. Clinical challenges Making a clinical ante-mortem diagnosis of LRTI in a cat can be challenging. Consistent historical, clinical, haematological and radiographic abnormalities are often lacking and findings may be non-specific. Astute clinical acumen, thorough investigation and high quality laboratory analysis are usually required for a diagnosis. Bronchoalveolar lavage, if feasible, and tests for lungworm should be routine in cats with lower respiratory tract signs. Lung fine needle aspiration may be useful in cases of diffuse or nodular pulmonary disease. Histopathology is rarely employed in ante-mortem investigations. Evidence base The authors have reviewed a substantial body of literature to provide information on many of the reported bacterial, parasitic, fungal and viral pathogens, including some that occur in Asia. Attention has been given to specific therapy for each pathogen, with evidence-based comments when there is a deviation from routine recommendations.


title: Chloroquine, a FDA-approved Drug, Prevents Zika Virus Infection and its Associated Congenital Microcephaly in Mice

text: Zika virus (ZIKV) has become a global public health emergency due to its rapidly expanding range and its ability to cause severe congenital defects such as microcephaly. However, there are no FDA-approved therapies or vaccines against ZIKV infection. Through our screening of viral entry inhibitors, we found that chloroquine (CQ), a commonly used antimalarial and a FDA-approved drug that has also been repurposed against other pathogens, could significantly inhibit ZIKV infection in vitro, by blocking virus internalization. We also demonstrated that CQ attenuates ZIKV-associated morbidity and mortality in mice. Finally, we proved that CQ protects fetal mice from microcephaly caused by ZIKV infection. Our methodology of focusing on previously identified antivirals in screens for effectiveness against ZIKV proved to be a rapid and efficient means of discovering new ZIKV therapeutics. Selecting drugs that were previously FDA-approved, such as CQ, also improves the likelihood that they may more quickly reach stages of clinical testing and use by the public.


Assessing the pandemic potential of emerging influenza


title: Evaluation of indirect immunofluorescence antibody test and enzyme-linked immunosorbent assay for the diagnosis of infection by Leishmania infantum in clinically normal and sick cats

text: Abstract Cats that live in areas where canine and human leishmaniosis due to Leishmania infantum is endemic may become infected and may develop anti-Leishmania antibodies. In this study 50 clinically normal and 50 cats with cutaneous and/or systemic signs that lived in an endemic area and had been previously examined for infection by L. infantum using PCR in four different tissues were serologically tested for the presence of anti-Leishmania IgG (IFAT and ELISA) and IgM (IFAT). The aim was to compare the results of IFAT, ELISA and PCR and to investigate the possible associations between seropositivity to Leishmania spp and signalment, living conditions, season of sampling, health status of the cats, and seropositivity to other infectious agents. Low concentrations of anti-Leishmania IgG were detected by IFAT in 10% of the cats and by ELISA in 1%, whereas anti-Leishmania IgM were detected by IFAT in 1%. There was disagreement between the results of IFAT and ELISA for anti-Leishmania IgG (P = 0.039) and between all serological tests and PCR (P < 0.001). The diagnostic sensitivity of all serological tests, using PCR as the gold standard, was very low, but ELISA and IFAT for anti-Leishmania IgM had 100% specificity. The diagnostic sensitivity of all serological tests could not be improved by changing the cut-off values. Seropositivity for Leishmania spp was not associated with signalment, living conditions, season of sampling and health status of the cats or with seropositivity to feline leukemia virus, feline immunodeficiency virus, feline coronavirus, Toxoplasma gondii and Bartonella henselae. In conclusion, because of their low sensitivity and very high specificity two of the evaluated serological tests (ELISA for anti-Leishmania IgG and IFAT for anti-Leishmania IgM) may be useless as population screening tests but valuable for diagnosing feline infection by L. infantum.


title: Who speaks for nursing? COVID‐19 highlighting gaps in leadership

text: In many areas of the world, the discipline and profession of nursing has been strengthened through the efforts of strong and effective leaders in education, research and clinical practice. The Seventy-second World Health Assembly designated 2020 as the International Year of the Nurse and the Midwife; not only to honour the 200th anniversary of the birth of Florence Nightingale, but also to recognize the contributions of nurses and midwives to the health and well-being of the world's population. Recently, on World Health Day, the World Health Organisation (WHO) released the State of the World's Nursing 2020 report providing data on nursing globally (WHO 2020). The report also provides a compelling case for development of the global nursing workforce, investment in nursing education, jobs, and leadership. However, operationalizing these recommendations will require investment and support at all levels of government and across all health jurisdictions.


title: Dengue Virus Reporter Replicon is a Valuable Tool for Antiviral Drug Discovery and Analysis of Virus Replication Mechanisms

text: Dengue, the most prevalent arthropod-borne viral disease, is caused by the dengue virus (DENV), a member of the Flaviviridae family, and is a considerable public health threat in over 100 countries, with 2.5 billion people living in high-risk areas. However, no specific antiviral drug or licensed vaccine currently targets DENV infection. The replicon system has all the factors needed for viral replication in cells. Since the development of replicon systems, transient and stable reporter replicons, as well as reporter viruses, have been used in the study of various virological aspects of DENV and in the identification of DENV inhibitors. In this review, we summarize the DENV reporter replicon system and its applications in high-throughput screening (HTS) for identification of anti-DENV inhibitors. We also describe the use of this system in elucidation of the mechanisms of virus replication and viral dynamics in vivo and in vitro.


title: Mycobacterium tuberculosis Sulfolipid-1 Activates Nociceptive Neurons and Induces Cough

text: Pulmonary tuberculosis, a disease caused by Mycobacterium tuberculosis (Mtb), manifests with a persistent cough as both a primary symptom and mechanism of transmission. The cough reflex can be triggered by nociceptive neurons innervating the lungs, and some bacteria produce neuron-targeting molecules. However, how pulmonary Mtb infection causes cough remains undefined, and whether Mtb produces a neuron-activating, cough-inducing molecule is unknown. Here, we show that an Mtb organic extract activates nociceptive neurons in vitro and identify the Mtb glycolipid sulfolipid-1 (SL-1) as the nociceptive molecule. Mtb organic extracts from mutants lacking SL-1 synthesis cannot activate neurons in vitro or induce cough in a guinea pig model. Finally, Mtb-infected guinea pigs cough in a manner dependent on SL-1 synthesis. Thus, we demonstrate a heretofore unknown molecular mechanism for cough induction by a virulent human pathogen via its production of a complex lipid.


title: Longitudinal surveillance of rotavirus A genotypes circulating in a high milk yield dairy cattle herd after the introduction of a rotavirus vaccine

text: Abstract Worldwide, neonatal diarrhea is one of the most important health issues affecting dairy calves, and rotavirus A (RVA) is one of its primary causes. Among the measures to mitigate the risk of diarrhea outbreaks, cow vaccination stands out as one of the most important. However, the immune pressure resulting from routine vaccination may be able to select specific G and P genotypes in RVA field strains. This study aimed to determine the frequency and intensity of neonatal diarrhea and the incidence of RVA and attempted to monitor the G and P genotypes present in the RVA strains circulating in a high milk yield cattle herd vaccinated with RVA G6P[5] strain. Fecal samples (n = 1220) from 122 Holstein heifer calves between 0–30 days old that were born from RVA-vaccinated cows were collected at 10 different time points, regardless of the presence or absence of diarrhea. The presence of RVA in fecal samples was determined by the polyacrylamide gel electrophoresis (PAGE) technique and confirmed by reverse transcription polymerase chain reaction (RT-PCR). G and P amplicons from 10 RVA-positive fecal samples from calves of different ages and collections were subjected to nucleotide sequencing. The proportion of the calves and fecal samples that were positive for RVA were 62.3% (76/122) and 8.1% (99/1220), respectively. Using sequence analysis, all 10 RVA field strains presented genotype G10P[11]. The protection of G6P[5] vaccination is clear, as this genotype was not detected in this study, and it is known that vaccination against RVA reduces the incidence of diarrhea independent of genotype involved. This result demonstrates the importance of epidemiological monitoring of RVA genotypes circulating in vaccinated dairy cattle herds to the early detection of new potential pathogenic RVA strains.


title: Acute peritoneal dialysis in the treatment of COVID-19-related acute kidney injury

text: The kidney is not typically the main target of severe acute respiratory syndrome coronavirus 2, but surprisingly, acute kidney injury (AKI) may occur in 4–23% of cases, whereas the dialysis management of AKI from coronavirus 2019 has not gained much attention. The severity of the pandemic has resulted in significant shortages in medical supplies, including respirators, ventilators and personal protective equipment. Peritoneal dialysis (PD) remains available and has been used in clinical practice for AKI for >70 years; however, it has been used on only a limited basis and therefore experience and knowledge of its use has gradually vanished, leaving a considerable gap. The turning point came in 2007, with a series of sequential publications providing solid evidence that PD is a viable option. As there was an availability constraint and a capacity limit of equipment/supplies in many countries, hemodialysis and convective therapies became alternatives. However, even these therapies are not available in many countries and their capacity is being pushed to the limit in many cities. Evidence-based PD experience lends support for the use of PD now.


title: Decrease and Delay in Hospitalization for Acute Coronary Syndromes during the 2020 SARS-CoV-2 Pandemic

text: The diffusion of SARS-CoV-2 forced the Italian population to restrictive measures that modified the patients’ response to non-SARS-CoV-2 medical conditions. We evaluated all ACS-patients admitted in three high-volume hospitals during the first month of SARS-CoV-2 Italian-outbreak and compared them to ACS-patients admitted during the same period 1-year before. Hospitalization for ACS decreased from 162 patients in 2019 vs 2020:84 patients in 2020. In 2020 both door-to-balloon and symptoms-to-PCI were longer and admission levels of hs-cTnI were higher. They had a lower discharged residual LV-function and an increased predicted late cardiovascular mortality based on their GRACE score.


title: Adult and pediatric pharyngitis: A review

text: Abstract Acute pharyngitis is frequently encountered in the ambulatory care setting. Although usually of viral etiology, streptococcal disease is the focus of diagnostic efforts, in light of significant suppurative and nonsuppurative sequelae. The traditional symptoms of fever, adenopathy, and pharyngeal exudate are suggestive, but not diagnostic of streptococcal pharyngitis. Thus, the importance of diagnostic testing, including Group A beta hemolytic strep antigen screen and culture, is emphasized. Recent innovations in therapy include modification of antibiotic dosing regimens and use of cephalosporins to improve patient compliance.


title: Quantitative determination of lymphocyte ACTH1–39

text: Abstract A method was developed for quantitative measurement of ACTH1–39, produced by human lymphocytes. It was shown that pH adjustment to 2 was essential for processing of the precursor molecule proopiomelanocortin (POMC). Linearity between time of incubation and ACTH production was shown. The existence of specific endopeptidases in lymphocytes for processing of the POMC molecule remains doubtful.


title: Recent progress on the diagnosis of 2019 Novel Coronavirus

text: Coronavirus disease 2019 (COVID‐19) caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has become a global pandemic. Therefore, convenient, timely and accurate detection of SARS‐CoV‐2 is urgently needed. Here, we review the types, characteristics and shortcomings of various detection methods, as well as perspectives for the SARS‐CoV‐2 diagnosis. Clinically, nucleic acid‐based methods are sensitive but prone to false‐positive. The antibody‐based method has slightly lower sensitivity but higher accuracy. Therefore, it is suggested to combine the two methods to improve the detection accuracy of COVID‐19.


Chinese People's Liberation Army on Action of Fighting against Ebola in Africa: Implications and Challenges


title: Seroprevalence of porcine respiratory coronavirus infection in Spanish breeding sows

text: Abstract The seroprevalence of porcine respiratory coronavirus (PRCV) among breeding sows was estimated in Castilla y León, the largest region in Spain, by a cross-sectional study. Serum samples (1247) were taken from sows from 58 different herds for this purpose throughout 1988 and were tested by a monoclonal antibody-capture ELISA (MACELISA), a test which detects antibodies to both transmissible gastroenteritis virus (TGEV) and PRCV. In order to discriminate positive sera to both coronaviruses, a blocking inhibition ELISA was used for all MACELISA-positive sera. By MACELISA, 31.4% of sera tested were positive and all of them were confirmed specifically as PRCV-positive by blocking inhibition ELISA, thus ruling out the presence of TGEV in the sampled area: 64% of the farms had at least one PRCV-seropositive sow, indicating that the infection was widespread Farms were classified into three categories, according to size and management practices. No significant differences were found in the prevalence rates among the three farm types; there was, however, a highly significant correlation between increasing farm size and increasing within-farm seropositivity.


title: Lung Ultrasound in a COVID Pandemic ‐ Choosing wisely

text: This is an opinion piece on the role of POCUS in COVID‐19, with a focus on lung ultrasound. It is not an instructional essay. Crisis management in medicine has often been likened to crisis management in the aviation industry. The important difference between pilots and clinicians being that the clinician's life was not in imminent danger, should one fail. The clinician did not have the same emotional urgency as the pilot. The COVID‐19 pandemic has changed this, clinicians are now faced with the need to make urgent decisions whilst exposed to some personal risk. Whether to embrace POCUS and lung ultrasound during this pandemic is an important decision. Whilst there are clear advantages, poorly considered overzealous uptake it is not without hazard, opportunity cost and potential risk to patient and clinician.


title: A missense polymorphism in porcine interferon-γ cDNA affects antiviral activity of the protein variant

text: Abstract We determined the interferon-γ (IFN-γ) cDNA sequence from three porcine breeds, Duroc, Landrance/Duroc hybrid, and Landrance breeds. Five single nucleotide polymorphisms (SNPs) of porcine IFN-γ (PoIFN-γ) were identified, respectively, at positions 269 (A/G), 376 (C/T), 426 (T/C), and 465 (T/C) of the coding sequence in Landrance/Duroc hybrid, and at position 251 (A/G) in Landrance breed. Among them, A269G and A251G polymorphisms resulted in Q67R and K61R replacements in the mature protein. PoIFN-γ cDNAs of Duroc breed (PoIFN-γ-W) and Landrance/Duroc hybrid (PoIFN-γ-M), which, respectively, encoded Q67 and R67, were introduced into a prokaryotic expression vector pET32 to express recombinant PoIFN-γ-W (rPoIFN-γ-W) and rPoIFN-γ-M protein variants in Escherichia coli. The identity of both protein variants was further confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We then compared bioactivities of these two recombinant proteins. Although both recombinant protein variants exhibited comparable activities in antiproliferation of PK-15 cells and in nitric oxide (NO) induction of porcine peripheral monocytes, antiviral activity of rPoIFN-γ-W protein was significantly higher (P <0.001) than that of rPoIFN-γ-M protein in a plaque inhibition assay using pseudorabies virus (PRV). IC50 values of rPoIFN-γ-W and rPoIFN-γ-M protein in anti-PRV assay were determined as 5.3±1.3 and 9.3±4.3nM, respectively. In conclusion, we have identified five novel SNPs in PoIFN-γ cDNA, including two missense polymorphisms that result in Q67R and K61R replacements. Our results further demonstrate that Q67R can markedly reduce antiviral activity of the PoIFN-γ protein. This is the first report that shows the functional SNP in the coding region of IFN-γ. In the future, it is imperative to determine whether Q67R replacement in IFN-γ may have disease association.


title: Chapter sixteen Peptide Nanomedicines for Treatment of Acute Lung Injury

text: Abstract Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) represent a heterogenous group of lung disease in critically ill patients. Despite the increased understanding of the molecular pathogenesis of ARDS, the mortality remains unacceptably high, ranging from 34% to 64%. Hence, ARDS represents an unmet medical need with an urgency to develop effective pharmacotherapies. Several promising targets that have been identified as potential therapies for ARDS have been limited because of difficulty with delivery. In particular, delivery of peptides and proteins to the lung is an ongoing challenge. Nanobiotechnology and nanoscience are the basis of innovative techniques to deliver drugs targeted to the site of inflamed organs, such as the lungs. Nanoscale drug delivery systems have the ability to improve the pharmacokinetics and pharmakodynamics of agents allowing an increase in the biodistribution of therapeutic agents to target organs, resulting in improved efficacy with reduction in drug toxicity. These systems are exploited for therapeutic purpose to carry the drug in the body in a controlled manner from the site of administration to the therapeutic target. Hence, it is an attractive strategy to test potential targets for ALI/ARDS using nanotechnology. To this end, we have identified several potential targets and proposed the delivery of these agents using nanomicelles to improve the drug delivery.


Killed in action: Microbiologists and clinicians as victims of their occupation Part 4: Tick-borne Relapsing Fever, Malta Fever, Glanders, SARS


title: Pandemic preparedness and responses: WHO to turn to in a crisis?

text: The PLOS Medicine editors discuss the role of the World Health Organization in pandemic responses.


title: Viral channel forming proteins — How to assemble and depolarize lipid membranes in silico()

text: Viral channel forming proteins (VCPs) have been discovered in the late 70s and are found in many viruses to date. Usually they are small and have to assemble to form channels which depolarize the lipid membrane of the host cells. Structural information is just about to emerge for just some of them. Thus, computational methods play a pivotal role in generating plausible structures which can be used in the drug development process. In this review the accumulation of structural data is introduced from a historical perspective. Computational performances and their predictive power are reported guided by biological questions such as the assembly, mechanism of function and drug–protein interaction of VCPs. An outlook of how coarse grained simulations can contribute to yet unexplored issues of these proteins is given. This article is part of a Special Issue entitled: Membrane Proteins edited by J.C. Gumbart and Sergei Noskov.


title: Rhinovirus species and clinical characteristics in the first wheezing episode in children

text: The clinical data on the first wheezing episodes induced by different rhinovirus (RV) species are still limited. We aimed to investigate the prevalence of RV genotypes, sensitization status and clinical characteristics of patients having a respiratory infection caused by either different RV species or other respiratory viruses. The study enrolled 111 patients (aged 3–23 months, 79% hospitalized, 76% with RV infection) with the first wheezing episode. RV-specific sequences were identified by partial sequencing of VP4/VP2 and 5′ non-coding regions with 80% success rate. The investigated clinical and laboratory variables included atopic characteristics and illness severity, parental atopic illnesses, and parental smoking. Of the study children, 56% percent had ≥1 atopic characteristic (atopy, eczema and/or blood eosinophil count ≥0.4×10(9)/L) and 23% were sensitised to allergens. RV-C was detected in 58% of RV positive samples, followed by RV-A (20%) and RV-B (1.2%). Children with RV-A and RV-C induced wheezing were older (p = 0.014) and had more atopic characteristics (p = 0.001) than those with non-RV. RV-A and RV-C illnesses had shorter duration of preadmission symptoms and required more bronchodilator use at the ward than non-RV illnesses (both p < 0.05, respectively) RV-C is the most common cause of severe early wheezing. Atopic and illness severity features are associated with children having RV-A or RV-C induced first wheezing episode rather than with children having a non-RV induced wheezing.


title: The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model

text: BACKGROUND: Oral immunization with vaccines may be an effective strategy for prevention of Clostridium difficile infection (CDI). However, application of previously developed vaccines for preventing CDI has been limited due to various reasons. Here, we developed a recombinant Lactococcus lactis oral vaccine and evaluated its effect on a C. difficile-infected animal model established in golden hamsters in attempt to provide an alternative strategy for CDI prevention. METHODS: Recombinant L. lactis vaccine was developed using the pTRKH2 plasmid, a high-copy-number Escherichia coli-L. shuttle vector: 1) L. lactis expressing secreted proteins was constructed with recombinant pTRKH2 (secreted-protein plasmid) carrying the Usp45 signal peptide (SPUsp45), nontoxic adjuvanted tetanus toxin fragment C (TETC), and 14 of the 38 C-terminal repeats (14CDTA) of nontoxic C. difficile toxin A (TcdA); and 2) L. lactis expressing secreted and membrane proteins was constructed with recombinant pTRKH2 (membrane-anchored plasmid) carrying SPUsp45, TETC, 14CDTA, and the cell wall-anchored sequence of protein M6 (cwaM6). Then, 32 male Syrian golden hamsters were randomly divided into 4 groups (n = 8 each) for gavage of normal saline (blank control) and L. lactis carrying the empty shuttle vector, secreted-protein plasmid, and membrane-anchored plasmid, respectively. After 1-week gavage of clindamycin, the animals were administered with C. difficile spore suspension. General symptoms and intestinal pathological changes of the animals were examined by naked eye and microscopy, respectively. Protein levels of anti-TcdA IgG/IgA antibodies in intestinal tissue and fluid were analyzed by enzyme-linked immunosorbent assay (ELISA). A cell culture cytotoxicity neutralization assay was done by TcdA treatment with or without anti-TcdA serum pre-incubation or treatment. Apoptosis of intestinal epithelial cells was examined by flow cytometry (FL) assay. Expression of mucosal inflammatory cytokines in the animals was detected by polymer chain reaction (PCR) assay. RESULTS: After the C. difficile challenge, the animals of control group had severe diarrhea symptoms on day 1 and all died on day 4, indicating that the CDI animal model was established in hamster. Of the 3 immunization groups, secreted-protein and membrane-anchored plasmid groups had significantly lower mortalities, body weight decreases, and pathological scores, with higher survival rate/time than the empty plasmid group (P < 0.05). The tilter of IgG antibody directed against TcdA was significantly higher in serum and intestinal fluid of secreted-protein and membrane-anchored plasmid groups than in the empty plasmid group (P < 0.05) while the corresponding titer of IgA antibody directed against TcdA had no substantial differences (P > 0.05). The anti-TcdA serum of membrane-anchored plasmid group neutralized the cytotoxicity of 200 ng/ml TcdA with the best protective effect achieved by anti-TcdA serum pre-incubation. The incidences of TcdA-induced death and apoptosis of intestinal epithelial cells were significantly reduced by cell pre-incubation or treatment with anti-TcdA serum of membrane-anchored plasmid group (P < 0.05). MCP-1, ICAM-1, IL-6, and Gro-1 mRNA expression levels were the lowest in cecum tissue of the membrane-anchored groups compared to the other groups. CONCLUSION: Recombinant L. lactis live vaccine is effective for preventing CDI in the hamster model, thus providing an alternative for immunization of C. difficile-associated diseases.


title: Pandemic H1N1 in Canada and the use of evidence in developing public health policies – A policy analysis

text: Abstract When responding to a novel infectious disease outbreak, policies are set under time constraints and uncertainty which can limit the ability to control the outbreak and result in unintended consequences including lack of public confidence. The H1N1 pandemic highlighted challenges in public health decision-making during a public health emergency. Understanding this process to identify barriers and modifiable influences is important to improve the response to future emergencies. The purpose of this study is to examine the H1N1 pandemic decision-making process in Canada with an emphasis on the use of evidence for public health decisions. Using semi-structured key informant interviews conducted after the pandemic (July–November 2010) and a document analysis, we examined four highly debated pandemic policies: use of adjuvanted vaccine by pregnant women, vaccine priority groups and sequencing, school closures and personal protective equipment. Data were analysed for thematic content guided by Lomas' policy decision-making framework as well as indicative coding using iterative methods. We interviewed 40 public health officials and scientific advisors across Canada and reviewed 76 pandemic policy documents. Our analysis revealed that pandemic pre-planning resulted in strong beliefs, which defined the decision-making process. Existing ideological perspectives of evidence strongly influenced how information was used such that the same evidentiary sources were interpreted differently according to the ideological perspective. Participants recognized that current models for public health decision-making failed to make explicit the roles of scientific evidence in relation to contextual factors. Conflict avoidance theory explained policy decisions that went against the prevailing evidence. Clarification of roles and responsibilities within the public health system would reduce duplication and maintain credibility. A more transparent and iterative approach to incorporating evidence into public health decision-making that reflects the realities of the external pressures present during a public health emergency is needed.


title: Disaster Competency Development and Integration in Nursing Education

text: Nurses, because of their nursing education and perspective practicing in multiple roles and settings, are uniquely qualified for mass casualty preparedness and response. Educating the current 2.7 million registered nurses and all future nursing graduates is a daunting task. Nursing education must ensure that graduates are prepared with the necessary knowledge and skills for mass casualty incidents. Four key entities are essential for education's successful implementation of disaster preparedness: education and professional organizations, accreditation and regulatory bodies, schools of nursing, and continuing education providers. This article examines the role each of these key entities plays in the development of a nursing workforce prepared for mass casualty response. In addition, the International Nursing Coalition for Mass Casualty Education (INCMCE) registered nurse (RN) competencies for mass casualty incidents and guidelines for integrating these competencies into the nursing education curricula are presented.


title: Evolution of the 5′-end of genomic rna of murine coronaviruses during passages in vitro

text: Abstract The 5′-ends of the genomic RNA and subgenomic mRNAs of murine coronavirus (MHV) have a stretch of approximately 70 nucleotides of leader sequences. The 3'-region of this leader sequence contains several repeats of a pentanucleotide (UCUAA), whose number varies among different MHV strains. It has been demonstrated that this UCUAA repeat plays crucial roles in the discontinuous transcription of MHV mRNAs. In the present study, we demonstrate that the number of UCUAA repeats in the leader sequence of MHV genome rapidly decreases during serial passages of viruses on susceptible cells. The downward evolution of the number of UCUAA repeats was not due to a higher growth rate of the viruses with fewer repeats, but seemed to be due to homologous interference between viruses with different numbers of UCUAA repeat. The ease with which these variant viruses arose suggests the high frequency of the occurrence of this deletion during RNA replication. This finding is in agreement with the proposed discontinuous and nonprocessive mode of coronavirus RNA synthesis. Analysis of the intracellular subgenomic mRNA species of viruses with different numbers of UCUAA repeats and of MHV recombinant viruses suggests that the number of this pentanucleotide repeat at the 3′-end of the leader sequence may regulate the synthesis of certain mRNA species, in agreement with the leader-primed transcription mechanism.


title: Development and evaluation of an immunochromatographic strip for rapid detection of porcine hemagglutinating encephalomyelitis virus

text: BACKGROUND: The incidence of PHE among pigs in many countries is on the rise, and it has caused great economic losses to the pig industry. Therefore, the development of a sensitive, specific, and easily-performed assay is crucial for the rapid detection and surveillance of PHE-CoV infection and transmission. RESULTS: An immunochromatographic strip was developed for the detection of PHE-CoV. The colloidal gold-labeled MAb 4D4 was used as the detection reagent, and the MAb 1E2 and goat anti-mouse IgG coated the strip's test and control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHE-CoV with a HA unit of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity or specificity. Using RT-PCR as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. There was an excellent agreement between the results obtained by RT-PCR and the immunochromatographic strips (kappa = 0.976). Additionally, there was a strong agreement between the sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatographic strips (Kappa = 0.976). When the immunochromatographic strips were used for diagnosing PHE-CoV infection in the Jilin Province, the PHE-CoV-positive rate ranged from 61.54% in the Jilin district to 17.95% in the Songyuan district. CONCLUSIONS: Based on its high specificity, sensitivity, and stability, the immunochromatographic strip would be suitable for on-site detection of PHE-CoV for surveillance and epidemiological purposes.


title: Olfactory testing does not predict β-amyloid, MRI measures of neurodegeneration or vascular pathology in the British 1946 birth cohort

text: OBJECTIVE: To explore the value of olfactory identification deficits as a predictor of cerebral β-amyloid status and other markers of brain health in cognitively normal adults aged ~ 70 years. METHODS: Cross-sectional observational cohort study. 389 largely healthy and cognitively normal older adults were recruited from the MRC National Survey of Health and Development (1946 British Birth cohort) and investigated for olfactory identification deficits, as measured by the University of Pennsylvania Smell Identification Test. Outcome measures were imaging markers of brain health derived from 3 T MRI scanning (cortical thickness, entorhinal cortex thickness, white matter hyperintensity volumes); (18)F florbetapir amyloid-PET scanning; and cognitive testing results. Participants were assessed at a single centre between March 2015 and January 2018. RESULTS: Mean (± SD) age was 70.6 (± 0.7) years, 50.8% were female. 64.5% had hyposmia and 2.6% anosmia. Olfaction showed no association with β-amyloid status, hippocampal volume, entorhinal cortex thickness, AD signature cortical thickness, white matter hyperintensity volume, or cognition. CONCLUSION AND RELEVANCE: In the early 70s, olfactory function is not a reliable predictor of a range of imaging and cognitive measures of preclinical AD. Olfactory identification deficits are not likely to be a useful means of identifying asymptomatic amyloidosis. Further studies are required to assess if change in olfaction may be a proximity marker for the development of cognitive impairment. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00415-020-10004-4) contains supplementary material, which is available to authorized users.


title: [Preliminary study of the relationship between novel coronavirus pneumonia and liver function damage: a multicenter study].

text: Objective: To analyze the clinical characteristics of cases of novel coronavirus pneumonia and a preliminary study to explore the relationship between different clinical classification and liver damage. Methods: Consecutively confirmed novel coronavirus infection cases admitted to seven designated hospitals during January 23, 2020 to February 8, 2020 were included. Clinical classification (mild, moderate, severe, and critical) was carried out according to the diagnosis and treatment program of novel coronavirus pneumonia (Trial Fifth Edition) issued by the National Health Commission. The research data were analyzed using SPSS19.0 statistical software. Quantitative data were expressed as median (interquartile range), and qualitative data were expressed as frequency and rate. Results: 32 confirmed cases that met the inclusion criteria were included. 28 cases were of mild or moderate type (87.50%), and four cases (12.50%) of severe or critical type. Four cases (12.5%) were combined with one underlying disease (bronchial asthma, coronary heart disease, malignant tumor, chronic kidney disease), and one case (3.13%) was simultaneously combined with high blood pressure and malignant tumor. The results of laboratory examination showed that the alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), and total bilirubin (TBil) for entire cohort were 26.98 (16.88 ~ 46.09) U/L and 24.75 (18.71 ~ 31.79) U/L, 39.00 (36.20 ~ 44.20) g/L and 16.40 (11.34- ~ 21.15) mmol/L, respectively. ALT, AST, ALB and TBil of the mild or moderate subgroups were 22.75 (16.31- ~ 37.25) U/L, 23.63 (18.71 ~ 26.50) U/L, 39.70 (36.50 ~ 46.10) g/L, and 15.95 (11.34 ~ 20.83) mmol/L, respectively. ALT, AST, ALB and TBil of the severe or critical subgroups were 60.25 (40.88 ~ 68.90) U/L, 37.00 (20.88 ~ 64.45) U/L, 35.75 (28.68 ~ 42.00) g/L, and 20.50 (11.28 ~ 25.00) mmol/L, respectively. Conclusion: The results of this multicenter retrospective study suggests that novel coronavirus pneumonia combined with liver damage is more likely to be caused by adverse drug reactions and systemic inflammation in severe patients receiving medical treatment. Therefore, liver function monitoring and evaluation should be strengthened during the treatment of such patients.


title: Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)

text: Abstract Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A540. The neutralizing antibody titers can be easily determined with either of the two new assays. In this report, we described the utility of these two new neutralization assays in measuring the neutralizing activities against SCV infection from rabbit sera immunized with various forms of spike protein of SCV.


title: In vitro and in vivo identification of a novel cytotoxic T lymphocyte epitope from Rv3425 of Mycobacterium tuberculosis

text: The identification of novel cytotoxic T lymphocyte (CTL) epitopes is important to analysis of the involvement of CD8(+) T cells in Mycobacterium tuberculosis infection as well as to the development of peptide vaccines. In this study, a novel CTL epitope from region of difference 11 encoded antigen Rv3425 was identified. Epitopes were predicted by the reversal immunology approach. Rv3425‐p118 (LIASNVAGV) was identified as having relatively strong binding affinity and stability towards the HLA‐A*0201 molecule. Peripheral blood mononuclear cells pulsed by this peptide were able to release interferon‐γ in healthy donors (HLA‐A*02(+) purified protein derivative(+)). In cytotoxicity assays in vitro and in vivo, Rv3425‐p118 induced CTLs to specifically lyse the target cells. Therefore, this epitope could provide a subunit component for designing vaccines against Mycobacterium tuberculosis.


title: Implications for Neuromodulation Therapy to Control Inflammation and Related Organ Dysfunction in COVID-19

text: COVID-19 is a syndrome that includes more than just isolated respiratory disease, as severe acute respiratory syndrome–coronavirus 2 (SARS-CoV2) also interacts with the cardiovascular, nervous, renal, and immune system at multiple levels, increasing morbidity in patients with underlying cardiometabolic conditions and inducing myocardial injury or dysfunction. Emerging evidence suggests that patients with the highest rate of morbidity and mortality following SARS-CoV2 infection have also developed a hyperinflammatory syndrome (also termed cytokine release syndrome). We lay out the potential contribution of a dysfunction in autonomic tone to the cytokine release syndrome and related multiorgan damage in COVID-19. We hypothesize that a cholinergic anti-inflammatory pathway could be targeted as a therapeutic avenue. [Figure: see text]


title: Severe Acute Respiratory Syndrome (SARS)

text: This article reviews the virology, history, pathology, epidemiology, clinical presentations, complications, radiology, laboratory testing, diagnosis, treatment, and prevention of severe respiratory distress syndrome, with reference to documented outbreaks of the disease.


title: Molecular viral epidemiology and clinical characterization of acute febrile respiratory infections in hospitalized children in Taiwan

text: Acute respiratory infection (ARI) is a leading cause of morbidity and hospitalization in children. To profile the viruses causing ARI in children admitted to a community‐based hospital in central Taiwan, a cross‐sectional study was conducted on children under 14 years of age that were hospitalized with febrile ARI. Viral etiology was determined using conventional cell culture and a commercial respiratory virus panel fast assay (xTAG RVP), capable of detecting 19 different respiratory viruses and subtype targets. Demographic, clinical, and laboratory data were recorded and analyzed. The RVP fast assay identified at least one respiratory virus in 130 of the 216 specimens examined (60.2%) and rose to 137 (63.4%) by combining the results of cell culture and RVP fast assay. In order of frequency, the etiological agents identified were, rhinovirus/enterovirus (24.6%), respiratory syncytial virus (13.8%), adenovirus (11.5%), parainfluenza virus (9.2%), influenza B (8.4%), influenza A (5.4%), human metapneumovirus (4.6%), human coronavirus (2%), and human bocavirus (2%). Co‐infection did not result in an increase in clinical severity. The RVP assay detected more positive specimens, but failed to detect 6 viruses identified by culture. The viral detection rate for the RVP assay was affected by how many days after admission the samples were taken (P = 0.03). In conclusion, Rhinovirus/enterovirus, respiratory syncytial virus, and adenovirus were prevalent in this study by adopting RVP assay. The viral detection rate is influenced by sampling time, especially if the tests are performed during the first three days of hospitalization. J. Med. Virol. 87:1860–1866, 2015. © 2015 Wiley Periodicals, Inc.


title: Guidance on breastfeeding during the Covid-19 pandemic

text: OBJECTIVE: These recommendations aim to provide guidance on breastfeeding for mothers with suspected or confirmed Covid-19 METHODS: We performed a review of the recent medical literature on breastfeeding mothers with suspected or confirmed Covid-19, focusing on the neonatal period RESULTS: We analyzed 20 recent publications on breastfeeding, Covid-19, and its transmission through breastmilk We presented possible options for breastfeeding and their consequences for the mother and the child CONCLUSION: All maternal decisions in relation to breastfeeding are justifiable since the infection by Covid-19 is still poorly known However, puerperal women and their families must be very well informed to make a conscious choice based on the information available in the literature so far


title: Saposhnikoviae divaricata: a phytochemical, pharmacological, and pharmacokinetic review

text: Abstract Saposhnikoviae divaricata (Turcz.) Schischk (SD) is a traditional Chinese herb commonly used to treat clinical conditions such as rheumatism and allergic rhinitis. This review article evaluates a collection of works on in vitro and biochemical studies of SD. The discourse on the diverse class of chromones and coumarins in SD offers an insight to the pharmacological effects of these bioactive constituents as anti-inflammatory, analgesic, immunoregulatory, antioxidative, and anti-proliferative agents. It is highlighted that there is a structural relationship between the constituents and bioactive activities, which in effect provides a valid reasoning and reaffirm the use of SD in the treatment of the pathologies in Chinese medicine.


title: Antibody and cytokine responses in kittens during the development of feline infectious peritonitis (FIP)

text: Abstract Two recombinant FIPV spike proteins were assessed for their immunogenic properties in 8-week-old kittens, which were then challenged intranasally with FIPV 79-1146. Humoral responses were assessed by ELISA and serum neutralisation test. Changes in PBMC cytokine mRNA levels were detected by a reverse transcription, semiquantitative polymerase chain reaction assay (RT-sqPCR), assessing IL-2, IL-4, IL-6, IL-10, IL-12 and IFNγ. All of the kittens developed clinical signs typical of FIP, which were confirmed on gross post mortem examination. The recombinant proteins induced little or no specific antibody response prior to challenge, and failed to alter the course of disease compared to controls. One week after virus challenge, the stimulated PBMCs showed small increases in the expression of IL-6 and IFNγ mRNA, which correlated with a transient pyrexia. After this time expression of IL-6 mRNA remained unaltered but, as FIP developed, mRNA levels of IL-2, IL-4, IL-10, IL-12 and IFNγ became markedly depressed.


title: Aspectos Epidemiológicos, Clínica Y Mecanismos De Control De La Pandemia Por Sars-Cov-2: Situación En España

text: Resumen En enero de 2020, las autoridades chinas confirmaron el agente causante de un brote de casos con neumonía de etiología desconocida aparecido en diciembre de 2019. La Organización Mundial de la Salud (OMS) denominó COVID-19 a este nuevo virus de la familia coronavirus. El día 12 de marzo la OMS declaró este brote como pandemia al tener una propagación mundial. Las epidemias humanas suelen seguir al inicio un crecimiento exponencial, esto significa que a partir de un pequeño número de casos en pocos días se observa un gran incremento en el número de personas infectadas. Las manifestaciones clínicas y el espectro de la enfermedad por SARS-CoV-2 abarcan desde presentaciones leves a críticas. En los casos con una progresión fatal las complicaciones más graves llegan a ocasionar fallo multiorgánico y la muerte del paciente. El SARS-CoV-2 tiene una elevada tasa de transmisibilidad, la vía de transmisión entre humanos se da a través de las secreciones de personas infectadas, las manos o los fómites contaminados. En general, las medidas de control de la epidemia utilizadas hasta el momento se han basado en intervenciones adoptadas de la epidemiología clásica, identificando y aislando los casos, siguiendo a los contactos y estableciendo restricciones, incluyendo la cuarentena, cierre de los centros educativos, evitando eventos en los que se produzca aglomeraciones de personas, limitando la movilidad. Estas medidas han sido efectivas en China, el foco originario del brote. Abstract In January 2020, the Chinese authorities confirmed the causative agent of an outbreak of cases with pneumonia of unknown etiology that appeared in December 2019. The World Health Organization (WHO) called COVID-19 is a new virus of the coronavirus family. On March 12, the WHO declared this outbreak as a pandemic as it spread worldwide. Human epidemics usually follow the start of exponential growth, this means that from a small number of cases in a few days, a large increase in the number of infected people is observed. The clinical manifestations and spectrum of symptomatic disease range from moderate to critical presentations. In cases with fatal progression, the most serious complications sometimes are cause of a multi-organ failure and patient death. SARS-CoV-2 has a high transmission rate, the route of transmission between humans is through the secretions of infected people, hands or contaminated objects. In general, the epidemic control measures used so far have been based on adoption of interventions from the classical epidemiology, identifying and isolating the cases, following the contacts and establishing restrictions, including quarantine, closure of educational centers, avoiding events in which crowds of people occur, limiting mobility. These measures have been effective in China, the original focus of the outbreak.


title: Viral Load Distribution in SARS Outbreak

text: An unprecedented community outbreak of severe acute respiratory syndrome (SARS) occurred in the Amoy Gardens, a high-rise residential complex in Hong Kong. Droplet, air, contaminated fomites, and rodent pests have been proposed to be mechanisms for transmitting SARS in a short period. We studied nasopharyngeal viral load of SARS patients on admission and their geographic distribution. Higher nasopharyngeal viral load was found in patients living in adjacent units of the same block inhabited by the index patient, while a lower but detectable nasopharyngeal viral load was found in patients living further away from the index patient. This pattern of nasopharyngeal viral load suggested that airborne transmission played an important part in this outbreak in Hong Kong. Contaminated fomites and rodent pests may have also played a role.


title: Fibrinolysis Shutdown Correlates to Thromboembolic Events in Severe COVID-19 Infection

text: BACKGROUND: Coronavirus disease 2019 (COVID-19) predisposes patients to a prothrombotic state with demonstrated microvascular involvement. The degree of hypercoagulability appears to correlate with outcomes, however optimal criteria to assess for the highest risk patients for thrombotic events remain unclear; we hypothesized that deranged thromboelastography (TEG) measurements of coagulation would correlate with thromboembolic events. METHODS: Patients admitted to an intensive care unit with COVID-19 diagnoses that had TEG analyses performed were studied. Conventional coagulation assays, D-dimer levels, and viscoelastic parameters were analyzed using a receiver operating characteristic curve to predict thromboembolic outcomes and new onset renal failure. RESULTS: Forty-four patients with COVID-19 were included in the analysis. Derangements in coagulation laboratory values including elevated D-Dimer, fibrinogen, PT, and PTT were confirmed; viscoelastic parameters showed an elevated maximum amplitude and low lysis at 30 minutes. A complete lack of lysis of clot at 30 minutes was seen in 57% of patients and predicted VTE with an AUROC of .742 (p=0.021). A D-Dimer cutoff of 2600 ng/ml predicted need for dialysis with an AUROC of .779 (p=0.005). Overall, patients with no lysis of clot at 30 minutes and a D-Dimer of greater than 2600 ng/ml had a rate of VTE of 50% compared to 0% for patients with neither risk factor (p=0.008) and had a hemodialysis rate of 80% compared to 14% (p=0.004). CONCLUSIONS: Fibrinolysis shutdown, as evidenced by elevated D-Dimer and complete failure of clot lysis at 30 minutes on thromboelastography, predicts thromboembolic events and need for hemodialysis in critically ill patients with COVID-19. Further clinical trials are required to ascertain the need for early therapeutic anticoagulation or fibrinolytic therapy to address this state of fibrinolysis shutdown.


title: Association between ambient PM2.5 and emergency department visits for psychiatric emergency diseases

text: Abstract Background Whether or not short-term exposure to particulate matter <2.5 μm in diameter (PM2.5) increases the risk of psychiatric emergency diseases is unclear. Methods The study was performed in a metropolis from January 2015 to December 2016. The exposure was PM2.5, and the confounders were weather (temperature and humidity) and other pollutants (PM10, SO2, CO, O3, and NO2). The outcomes were emergency department (ED) visits with psychiatric disease codes (F00-F99 in ICD10 codes). General additive models were used for the statistical analysis to calculate the adjusted relative risks (ARRs) and 95% confidence intervals (95% CIs) for the daily number of ED visits with a lag of 1 to 3 days following a 10 μg/m3 increase in PM2.5. Results During the study period, a total of 67,561 ED visits for psychiatric diseases were identified and tested for association with PM2.5. Daily ED visits for all psychiatric diseases were not associated with PM2.5 in the model that was not adjusted for other pollutants. The ARR (95% CI) in the model adjusted for SO2 was 1.011 (1.002–1.021) by 10 μg/m3 of PM2.5 on Lag 1 for all psychiatric diseases (F00-F99). The ARR (95% CI) in the model adjusted for O3 was 1.015 (1.003–1.029) by 10 μg/m3 of PM2.5 on Lag 1 for F40-F49 (Neurotic, stress-related and somatoform disorders). Conclusion An increase in PM2.5 showed a significant association with an increase in ED visits for all psychiatric diseases (F00-F99) and for neurotic, stress-related and somatoform disorders (F40-F49) on lag day 1.


title: “The life at the times of coronavirus” Microbes rule the world!

text: I used to begin my microbiology lecture series with a slogan “microbes rule the world”. And I continued by reminding the students that microbes, in fact, have been on earth billions of years before man and that they most probably stay here after mankind has been extinguished. The current SARS‐CoV‐2 pandemic makes this statement actual but also a bit creepy.


title: Broad Cross-Species Infection of Cultured Cells by Bat HKU2-Related Swine Acute Diarrhea Syndrome Coronavirus and Identification of Its Replication in Murine Dendritic Cells In Vivo Highlight Its Potential for Diverse Interspecies Transmission

text: Outbreaks of severe diarrhea in neonatal piglets in Guangdong, China, in 2017 resulted in the isolation and discovery of a novel swine enteric alphacoronavirus (SeACoV) derived from the species Rhinolophus bat coronavirus HKU2 (Y. Pan, X. Tian, P. Qin, B. Wang, et al., Vet Microbiol 211:15–21, 2017). SeACoV was later referred to as swine acute diarrhea syndrome CoV (SADS-CoV) by another group (P. Zhou, H. Fan, T. Lan, X.-L. Yang, et al., Nature 556:255–258, 2018). The present study was set up to investigate the potential species barriers of SADS-CoV in vitro and in vivo. We first demonstrated that SADS-CoV possesses a broad species tropism and is able to infect cell lines from diverse species, including bats, mice, rats, gerbils, hamsters, pigs, chickens, nonhuman primates, and humans. Trypsin contributes to but is not essential for SADS-CoV propagation in vitro. Furthermore, C57BL/6J mice were inoculated with the virus via oral or intraperitoneal routes. Although the mice exhibited only subclinical infection, they supported viral replication and prolonged infection in the spleen. SADS-CoV nonstructural proteins and double-stranded RNA were detected in splenocytes of the marginal zone on the edge of lymphatic follicles, indicating active replication of SADS-CoV in the mouse model. We identified that splenic dendritic cells (DCs) are the major targets of virus infection by immunofluorescence and flow cytometry approaches. Finally, we demonstrated that SADS-CoV does not utilize known CoV receptors for cellular entry. The ability of SADS-CoV to replicate in various cells lines from a broad range of species and the unexpected tropism for murine DCs provide important insights into the biology of this bat-origin CoV, highlighting its possible ability to cross interspecies barriers. IMPORTANCE Infections with bat-origin coronaviruses (CoVs) (severe acute respiratory syndrome CoV [SARS-CoV] and Middle East respiratory syndrome CoV [MERS-CoV]) have caused severe illness in humans after “host jump” events. Recently, a novel bat-HKU2-like CoV named swine acute diarrhea syndrome CoV (SADS-CoV) has emerged in southern China, causing lethal diarrhea in newborn piglets. It is important to assess the species barriers of SADS-CoV infection since the animal hosts (other than pigs and bats) and zoonotic potential are still unknown. An in vitro susceptibility study revealed a broad species tropism of SADS-CoV, including various rodent and human cell lines. We established a mouse model of SADS-CoV infection, identifying its active replication in splenic dendritic cells, which suggests that SADS-CoV has the potential to infect rodents. These findings highlight the potential cross-species transmissibility of SADS-CoV, although further surveillance in other animal populations is needed to fully understand the ecology of this bat-HKU2-origin CoV.


title: Accelerated viral dynamics in bat cell lines, with implications for zoonotic emergence

text: Bats host virulent zoonotic viruses without experiencing disease. A mechanistic understanding of the impact of bats’ virus hosting capacities, including uniquely constitutive immune pathways, on cellular-scale viral dynamics is needed to elucidate zoonotic emergence. We carried out virus infectivity assays on bat cell lines expressing induced and constitutive immune phenotypes, then developed a theoretical model of our in vitro system, which we fit to empirical data. Best fit models recapitulated expected immune phenotypes for representative cell lines, supporting robust antiviral defenses in bat cells that correlated with higher estimates for within-host viral propagation rates. In general, heightened immune responses limit pathogen-induced cellular morbidity, which can facilitate the establishment of rapidly-propagating persistent infections within-host. Rapidly-transmitting viruses that have evolved with bat immune systems will likely cause enhanced virulence following emergence into secondary hosts with immune systems that diverge from those unique to bats.


title: Preparation, quality criteria, and properties of human blood platelet lysate supplements for ex vivo stem cell expansion

text: Most clinical applications of human multipotent mesenchymal stromal cells (MSCs) for cell therapy, tissue engineering, regenerative medicine, and treatment of immune and inflammatory diseases require a phase of isolation and ex vivo expansion allowing a clinically meaningful cell number to be reached. Conditions used for cell isolation and expansion should meet strict quality and safety requirements. This is particularly true for the growth medium used for MSC isolation and expansion. Basal growth media used for MSC expansion are supplemented with multiple nutrients and growth factors. Fetal bovine serum (FBS) has long been the gold standard medium supplement for laboratory-scale MSC culture. However, FBS has a poorly characterized composition and poses risk factors, as it may be a source of xenogenic antigens and zoonotic infections. FBS has therefore become undesirable as a growth medium supplement for isolating and expanding MSCs for human therapy protocols. In recent years, human blood materials, and most particularly lysates and releasates of platelet concentrates have emerged as efficient medium supplements for isolating and expanding MSCs from various origins. This review analyzes the advantages and limits of using human platelet materials as medium supplements for MSC isolation and expansion. We present the modes of production of allogeneic and autologous platelet concentrates, measures taken to ensure optimal pathogen safety profiles, and methods of preparing PLs for MSC expansion. We also discuss the supply of such blood preparations. Produced under optimal conditions of standardization and safety, human platelet materials can become the future ‘gold standard’ supplement for ex vivo production of MSCs for translational medicine and cell therapy applications.


title: Recombinant infectious bronchitis virus (IBV) H120 vaccine strain expressing the hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) protects chickens against IBV and NDV challenge

text: Infectious bronchitis (IB) and Newcastle disease (ND) are common viral diseases of chickens, which are caused by infectious bronchitis virus (IBV) and Newcastle disease virus (NDV), respectively. Vaccination with live attenuated strains of IBV-H120 and NDV-LaSota are important for the control of IB and ND. However, conventional live attenuated vaccines are expensive and result in the inability to differentiate between infected and vaccinated chickens. Therefore, there is an urgent need to develop new efficacious vaccines. In this study, using a previously established reverse genetics system, we generated a recombinant IBV virus based on the IBV H120 vaccine strain expressing the haemagglutinin-neuraminidase (HN) protein of NDV. The recombinant virus, R-H120-HN/5a, exhibited growth dynamics, pathogenicity and viral titers that were similar to those of the parental IBV H120, but it had acquired hemagglutination activity from NDV. Vaccination of SPF chickens with the R-H120-HN/5a virus induced a humoral response at a level comparable to that of the LaSota/H120 commercial bivalent vaccine and provided significant protection against challenge with virulent IBV and NDV. In summary, the results of this study indicate that the IBV H120 strain could serve as an effective tool for designing vaccines against IB and other infectious diseases, and the generation of IBV R-H120-HN/5a provides a solid foundation for the development of an effective bivalent vaccine against IBV and NDV.


title: A Dimerization-Dependent Mechanism Drives the Endoribonuclease Function of Porcine Reproductive and Respiratory Syndrome Virus nsp11

text: Porcine reproductive and respiratory syndrome virus (PRRSV) RNA endoribonuclease nsp11 belongs to the XendoU superfamily and plays a crucial role in arterivirus replication. Here, we report the first crystal structure of the arterivirus nsp11 protein from PRRSV, which exhibits a unique structure and assembles into an asymmetric dimer whose structure is completely different from the hexameric structure of coronavirus nsp15. However, the structures of the PRRSV nsp11 and coronavirus nsp15 catalytic domains were perfectly superimposed, especially in the “active site loop” (His129 to His144) and “supporting loop” (Val162 to Thr179) regions. Importantly, our biochemical data demonstrated that PRRSV nsp11 exists mainly as a dimer in solution. Mutations of the major dimerization site determinants (Ser74 and Phe76) in the dimerization interface destabilized the dimer in solution and severely diminished endoribonuclease activity, indicating that the dimer is the biologically functional unit. In the dimeric structure, the active site loop and supporting loop are packed against one another and stabilized by monomer-monomer interactions. These findings may help elucidate the mechanism underlying arterivirus replication and may represent great potential for the development of antiviral drugs. IMPORTANCE Porcine reproductive and respiratory syndrome virus (PRRSV) is a member of the family Arteriviridae, order Nidovirales. PRRSV is a major agent of respiratory diseases in pigs, causing tremendous economic losses to the swine industry worldwide. The PRRSV nsp11 endoribonuclease plays a vital role in arterivirus replication, but its precise roles and mechanisms of action are poorly understood. Here, we report the first dimeric structure of the arterivirus nsp11 from PRRSV at 2.75-Å resolution. Structural and biochemical experiments demonstrated that nsp11 exists mainly as a dimer in solution and that nsp11 may be fully active as a dimer. Mutagenesis and structural analysis revealed NendoU active site residues, which are conserved throughout the order Nidovirales (families Arteriviridae and Coronaviridae) and the major determinants of dimerization (Ser74 and Phe76) in Arteriviridae. Importantly, these findings may provide a new structural basis for antiviral drug development.


title: Stochastic dynamics of an epidemic with recurrent spillovers from an endemic reservoir

text: Abstract Most emerging human infectious diseases have an animal origin. While zoonotic diseases originate from a reservoir, most theoretical studies have principally focused on single-host processes, either exclusively humans or exclusively animals, without considering the importance of animal to human transmission (i.e. spillover transmission) for understanding the dynamics of emerging infectious diseases. Here we aim to investigate the importance of spillover transmission for explaining the number and the size of outbreaks. We propose a simple continuous time stochastic Susceptible-Infected-Recovered model with a recurrent infection of an incidental host from a reservoir (e.g. humans by a zoonotic species), considering two modes of transmission, (1) animal-to-human and (2) human-to-human. The model assumes that (i) epidemiological processes are faster than other processes such as demographics or pathogen evolution and that (ii) an epidemic occurs until there are no susceptible individuals left. The results show that during an epidemic, even when the pathogens are barely contagious, multiple outbreaks are observed due to spillover transmission. Overall, the findings demonstrate that the only consideration of direct transmission between individuals is not sufficient to explain the dynamics of zoonotic pathogens in an incidental host.


title: Tertiary lymphoid organs in infection and autoimmunity

text: The lymph nodes (LNs) and spleen have an optimal structure that allows the interaction between T cells, B cells and antigen-presenting dendritic cells (DCs) on a matrix made up by stromal cells. Such a highly organized structure can also be formed in tertiary lymphoid organs (TLOs) at sites of infection or chronic immune stimulation. This review focuses on the molecular mechanisms of TLO formation and maintenance, the controversies surrounding the nature of the inducing events, and the functions of these structures in infection, transplantation and autoimmunity.


Pandemic vitality: on living and being alive in lockdown


title: First Results of a National External Quality Assessment Scheme for the detection of SARS-CoV-2 genome sequences

text: BACKGROUND: Broad and decentralised testing of SARS-CoV-2 RNA genomes is a WHOrecommended strategy to contain the SARS-CoV-2 pandemic by identifying infected cases in order to minimize onward transmission. With the need to increase the test capacities in Austria, nation-wide numerous laboratories rapidly implemented assays for molecular detection of SARSCoV-2 based on real-time RT-PCR assays. The objective of this study was to monitor reliability of the laboratory results for SARS-CoV-2 RNA detection through an external quality assessment (EQA) scheme. METHODS: For this, the Center for Virology, Medical University of Vienna was tasked by the Federal Ministry of Social Affairs, Health, Care and Consumer Protection to perform the first Austrian EQA on SARS-CoV-2 which was organised in cooperation with the Austrian Association for Quality Assurance and Standardization of Medical and Diagnostic Tests (ÖQUASTA). Data were analysed on the basis of qualitative outcome of testing in relation to the nucleic acid (NA) extraction and detection methods used. RESULTS AND CONCLUSION: A total of 52 laboratories participated, contributing results from 67 test panels comprising 42 distinct combinations of NA extraction and PCR reagents. By testing 3 positive (CT values: S1, 28.4; S2, 33.6; S3, 38.5) and 1 negative sample, no false-positive results were obtained by any of the laboratories. Otherwise, 40/67 tests (60%) detected all positive samples correctly as positive, but 25/67 tests (37%) did not detect the weakest positive sample (S3), and 3% reported S2 and S3 as falsenegative. Improvement in test sensitivity by focusing on NA extraction and/or PCR-based detection is recommended.


title: Nitazoxanide: A first-in-class broad-spectrum antiviral agent

text: Abstract Originally developed and commercialized as an antiprotozoal agent, nitazoxanide was later identified as a first-in-class broad-spectrum antiviral drug and has been repurposed for the treatment of influenza. A Phase 2b/3 clinical trial recently published in The Lancet Infectious Diseases found that oral administration of nitazoxanide 600mg twice daily for five days reduced the duration of clinical symptoms and reduced viral shedding compared to placebo in persons with laboratory-confirmed influenza. The same study also suggested a potential benefit for subjects with influenza-like illness who did not have influenza or other documented respiratory viral infection. From a chemical perspective, nitazoxanide is the scaffold for a new class of drugs called thiazolides. These small-molecule drugs target host-regulated processes involved in viral replication. Nitazoxanide is orally bioavailable and safe with extensive post-marketing experience involving more than 75 million adults and children. A new dosage formulation of nitazoxanide is presently undergoing global Phase 3 clinical development for the treatment of influenza. Nitazoxanide inhibits a broad range of influenza A and B viruses including influenza A(pH1N1) and the avian A(H7N9) as well as viruses that are resistant to neuraminidase inhibitors. It is synergistic with neuraminidase inhibitors, and combination therapy with oseltamivir is being studied in humans as part of ongoing Phase 3 clinical development. Nitazoxanide also inhibits the replication of a broad range of other RNA and DNA viruses including respiratory syncytial virus, parainfluenza, coronavirus, rotavirus, norovirus, hepatitis B, hepatitis C, dengue, yellow fever, Japanese encephalitis virus and human immunodeficiency virus in cell culture assays. Clinical trials have indicated a potential role for thiazolides in treating rotavirus and norovirus gastroenteritis and chronic hepatitis B and chronic hepatitis C. Ongoing and future clinical development is focused on viral respiratory infections, viral gastroenteritis and emerging infections such as dengue fever.


title: Une journée pour honorer les tout-petits partis trop tôt

text: La perte d’un tout-petit pendant la grossesse ou autour de la naissance provoque un état de détresse incommensurable. Cette souffrance est accentuée par l’incompréhension de l’entourage, par la solitude et par le silence qui règnent autour de ce drame, par le manque de reconnaissance de ces enfants et, pour les couples, par la difficulté de vivre le statut ambigu de parents sans enfants. Depuis 2013, un collectif associatif organise chaque année la journée “Une fleur, une vie”. Née d’une envie commune de créer un événement artistique et public, cette initiative permet à des centaines de personnes, en France et depuis l’étranger, d’honorer leur tout-petit et d’avancer à leur rythme dans leur deuil. A day to honour still-borns and victims of early infant death Losing a child during pregnancy or around birth causes immeasurable distress. This suffering is accentuated by the incomprehension of those around them, by the solitude and silence that surrounds this tragedy, by the lack of recognition of these children and, for couples, by the difficulty of living the ambiguous status of childless parents. Since 2013, an associative group has been organising the annual “One flower, one life” day. Born of a common desire to create an artistic and public event, this initiative allows hundreds of people, in France and from abroad, to honour their little ones and to move forward at their own pace in their mourning.


title: Cellular RNA Helicase DDX1 Is Involved in Transmissible Gastroenteritis Virus nsp14-Induced Interferon-Beta Production

text: Transmissible gastroenteritis virus (TGEV), an enteropathogenic coronavirus (CoV) of porcine, causes lethal watery diarrhea and severe dehydration in piglets and leads to severe economic losses in the swine industry. Unlike most CoVs that antagonize type I interferon (IFN) production, previous studies showed that TGEV infection induces IFN-I production both in vivo and in vitro. However, the underlying mechanism(s) remain largely unknown. In this study, we found that TGEV infection significantly facilitated IFN-β production as well as activation of the transcription factors IFN regulatory factor 3 (IRF3) and nuclear factor-kappaB (NF-κB) in porcine kidney (PK-15) cells. Screening of TGEV-encoded proteins demonstrated that non-structural protein 14 (nsp14) was the most potent IFN-β inducer and induced IFN-β production mainly by activating NF-κB but not IRF3. Further analysis showed that nsp14 interacted with DDX1, a member of the DExD/H helicase family. Knockdown of DDX1 by specific small interfering RNA (siRNA) significantly decreased nsp14-induced IFN-β production and NF-κB activation. Furthermore, TGEV-induced IFN-β production and IFN-stimulated gene (ISG) expression were decreased in cells transfected with DDX1-specific siRNA, indicating the vital role of DDX1 to TGEV-induced IFN-β responses. In summary, our data revealed a potential coactivator role of host RNA helicase DDX1 to the induction of IFN-β response initiated by TGEV and demonstrated that nsp14 is an important IFN inducer among the TGEV-encoded proteins.


title: COVID-19 Clinical Trials: A Teachable Moment for Improving Our Research Infrastructure and Relevance

text: The authors posit the need for rapid evaluation of therapies for COVID-19 as an inflection point spurring a much-needed rethinking of our research enterprise.


title: COVID-19, immune system response, hyperinflammation and repurposing antirheumatic drugs

text: In the Wuhan Province of China, in December 2019, the novel coronavirus 2019 (COVID-19) has caused a severe involvement of the lower respiratory tract leading to an acute respiratory syndrome. Subsequently, coronavirus 2 (SARS-CoV-2) provoked a pandemic which is considered a life-threatening disease. The SARS-CoV-2, a family member of betacoronaviruses, possesses single-stranded positive-sense RNA with typical structural proteins, involving the envelope, membrane, nucleocapsid and spike proteins that are responsible for the viral infectivity, and nonstructural proteins. The effectual host immune response including innate and adaptive immunity against SARS-Cov-2 seems crucial to control and resolve the viral infection. However, the severity and outcome of the COVID-19 might be associated with the excessive production of proinflammatory cytokines “cytokine storm” leading to an acute respiratory distress syndrome. Regretfully, the exact pathophysiology and treatment, especially for the severe COVID-19, is still uncertain. The results of preliminary studies have shown that immune-modulatory or immune-suppressive treatments such as hydroxychloroquine, interleukin (IL)-6 and IL-1 antagonists, commonly used in rheumatology, might be considered as treatment choices for COVID-19, particularly in severe disease. In this review, to gain better information about appropriate anti-inflammatory treatments, mostly used in rheumatology for COVID-19, we have focused the attention on the structural features of SARS-CoV-2, the host immune response against SARS-CoV-2 and its association with the cytokine storm.


title: Structural Determination of the Broadly Reactive Anti-IGHV1-69 Anti-idiotypic Antibody G6 and Its Idiotope

text: The heavy chain IGHV1-69 germline gene exhibits a high level of polymorphism and shows biased use in protective antibody (Ab) responses to infections and vaccines. It is also highly expressed in several B cell malignancies and autoimmune diseases. G6 is an anti-idiotypic monoclonal Ab that selectively binds to IGHV1-69 heavy chain germline gene 51p1 alleles that have been implicated in these Ab responses and disease processes. Here, we determine the co-crystal structure of humanized G6 (hG6.3) in complex with anti-influenza hemagglutinin stem-directed broadly neutralizing Ab D80. The core of the hG6.3 idiotope is a continuous string of CDR-H2 residues starting with M53 and ending with N58. G6 binding studies demonstrate the remarkable breadth of binding to 51p1 IGHV1-69 Abs with diverse CDR-H3, light chain, and antigen binding specificities. These studies detail the broad expression of the G6 cross-reactive idiotype (CRI) that further define its potential role in precision medicine.


title: Potential Inhibitors for Novel Coronavirus Protease Identified by Virtual Screening of 606 Million Compounds

text: The rapid outbreak of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in China followed by its spread around the world poses a serious global concern for public health. To this date, no specific drugs or vaccines are available to treat SARS-CoV-2 despite its close relation to the SARS-CoV virus that caused a similar epidemic in 2003. Thus, there remains an urgent need for the identification and development of specific antiviral therapeutics against SARS-CoV-2. To conquer viral infections, the inhibition of proteases essential for proteolytic processing of viral polyproteins is a conventional therapeutic strategy. In order to find novel inhibitors, we computationally screened a compound library of over 606 million compounds for binding at the recently solved crystal structure of the main protease (M(pro)) of SARS-CoV-2. A screening of such a vast chemical space for SARS-CoV-2 M(pro) inhibitors has not been reported before. After shape screening, two docking protocols were applied followed by the determination of molecular descriptors relevant for pharmacokinetics to narrow down the number of initial hits. Next, molecular dynamics simulations were conducted to validate the stability of docked binding modes and comprehensively quantify ligand binding energies. After evaluation of potential off-target binding, we report a list of 12 purchasable compounds, with binding affinity to the target protease that is predicted to be more favorable than that of the cocrystallized peptidomimetic compound. In order to quickly advise ongoing therapeutic intervention for patients, we evaluated approved antiviral drugs and other protease inhibitors to provide a list of nine compounds for drug repurposing. Furthermore, we identified the natural compounds (−)-taxifolin and rhamnetin as potential inhibitors of M(pro). Rhamnetin is already commercially available in pharmacies.


title: Operational considerations for peritoneal dialysis management during the COVID-19 pandemic

text: BACKGROUND: Coronavirus disease 2019 (COVID-19) is an infectious respiratory disease caused by a novel coronavirus—severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It is spread from human to human and has resulted in a global pandemic, posing a disastrous public health risk worldwide. Patients with chronic kidney disease, especially those on dialysis, are considered to be at higher risk of developing severe COVID-19 due to their immunocompromised status and frail condition. The home treatment setting of peritoneal dialysis (PD) has advantages in terms of implementing self-care when routine hospital visits and social activities are restricted, thus greatly reducing exposure of PD patients to the virus. METHODS AND RESULTS: We outline general operational considerations in PD management during the COVID-19 pandemic, including precautionary measures for PD patients and healthcare staff. Precautionary measures for PD patients include education on prevention of, and screening for, COVID-19, preclinic screening, in-clinic management, meticulous remote patient management and special hospitalization arrangements. The diagnosis and treatment of PD patients with COVID-19 are discussed. Precautionary measures for PD staff include continuous education on, and training in, COVID-19, exposure history surveillance and self-monitoring for COVID-19 among healthcare personnel, appropriate personal protective equipment and hand hygiene, organization of medical activities and staffing, and adequate environment cleaning. CONCLUSIONS: This is a battle of the entire human society against the novel coronavirus. Integrated teamwork among healthcare providers, supported by society as a whole, is needed as part of the ongoing public health response to try to slow the spread of COVID-19.


title: Sequence space coverage, entropy of genomes and the potential to detect non-human DNA in human samples

text: BACKGROUND: Genomes store information for building and maintaining organisms. Complete sequencing of many genomes provides the opportunity to study and compare global information properties of those genomes. RESULTS: We have analyzed aspects of the information content of Homo sapiens, Mus musculus, Drosophila melanogaster, Caenorhabditis elegans, Arabidopsis thaliana, Saccharomyces cerevisiae, and Escherichia coli (K-12) genomes. Virtually all possible (> 98%) 12 bp oligomers appear in vertebrate genomes while < 2% of 19 bp oligomers are present. Other species showed different ranges of > 98% to < 2% of possible oligomers in D. melanogaster (12–17 bp), C. elegans (11–17 bp), A. thaliana (11–17 bp), S. cerevisiae (10–16 bp) and E. coli (9–15 bp). Frequencies of unique oligomers in the genomes follow similar patterns. We identified a set of 2.6 M 15-mers that are more than 1 nucleotide different from all 15-mers in the human genome and so could be used as probes to detect microbes in human samples. In a human sample, these probes would detect 100% of the 433 currently fully sequenced prokaryotes and 75% of the 3065 fully sequenced viruses. The human genome is significantly more compact in sequence space than a random genome. We identified the most frequent 5- to 20-mers in the human genome, which may prove useful as PCR primers. We also identified a bacterium, Anaeromyxobacter dehalogenans, which has an exceptionally low diversity of oligomers given the size of its genome and its GC content. The entropy of coding regions in the human genome is significantly higher than non-coding regions and chromosomes. However chromosomes 1, 2, 9, 12 and 14 have a relatively high proportion of coding DNA without high entropy, and chromosome 20 is the opposite with a low frequency of coding regions but relatively high entropy. CONCLUSION: Measures of the frequency of oligomers are useful for designing PCR assays and for identifying chromosomes and organisms with hidden structure that had not been previously recognized. This information may be used to detect novel microbes in human tissues.