title: SRSF7 maintains its homeostasis through the expression of Split-ORFs and nuclear body assembly

text: SRSF7 is an essential RNA-binding protein whose misexpression promotes cancer. Here, we describe how SRSF7 maintains its protein homeostasis in murine P19 cells using an intricate negative feedback mechanism. SRSF7 binding to its premessenger RNA promotes inclusion of a poison cassette exon and transcript degradation via nonsense-mediated decay (NMD). However, elevated SRSF7 levels inhibit NMD and promote translation of two protein halves, termed Split-ORFs, from the bicistronic SRSF7-PCE transcript. The first half acts as dominant-negative isoform suppressing poison cassette exon inclusion and instead promoting the retention of flanking introns containing repeated SRSF7 binding sites. Massive SRSF7 binding to these sites and its oligomerization promote the assembly of large nuclear bodies, which sequester SRSF7 transcripts at their transcription site, preventing their export and restoring normal SRSF7 protein levels. We further show that hundreds of human and mouse NMD targets, especially RNA-binding proteins, encode potential Split-ORFs, some of which are expressed under specific cellular conditions.


title: Hemophagocytic syndrome and COVID-19

text: The case of a woman hospitalized due to COVID-19 is presented. The patient developed a severe macrophage activation syndrome diagnosed by bone marrow biopsy, which did not respond to immunoglobulin therapy.


O que é urgente e necessário para subsidiar as políticas de enfrentamento da pandemia de COVID-19 no Brasil?/ What is urgent and necessary to inform policies to deal with the COVID-19 pandemic in Brazil?


title: Isolation and molecular characterization of nephropathic infectious bronchitis virus isolates of Gujarat state, India

text: Infectious bronchitis (IB) is a common, highly contagious, acute, and economically important viral disease of chickens caused by Infectious bronchitis virus (IBV, sp. Avian coronavirus). Five pooled tissue suspensions of 50 layer birds and one reference Massachusetts vaccine strain were inoculated into specific pathogen free (SPF) chicken egg for isolation of IBV. Reverse-transcription polymerase chain reaction (RT-PCR) was carried out using post inoculated allontoic fluid to amplify the spike (S) glycoprotein of S1 subunit of IBV. All the eggs inoculated with five pooled tissue samples and vaccine sample showed dwarfing and curling of SPF embryos indicative of IBV. All the five samples and the vaccine sample produced the expected amplicons of 466 bp by RT-PCR. The sequencing of five isolates revealed that all the five sequences were 99.09–100 % similar among themselves and showed 99.10–100 % nucleotide identity with the vaccine strain. On multiple sequence alignment it was found that our isolates were more similar at S1 subunit nucleotide level with the reference Ma5 and H120 vaccine strains than the reference Mass41 strain. The sequences of Anand isolates revealed further genetic changes in the circulating IBV in comparison to previous isolate of Gujarat as well as higher differences with the strains isolated in other states showing substantial changes at genetic level in Indian IBV isolates, which may partially explain the increasing incidences of IB in the country in spite of the vaccination. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s13337-015-0248-x) contains supplementary material, which is available to authorized users.


title: Attenuation of a virulent swine acute diarrhea syndrome coronavirus strain via cell culture passage

text: Abstract Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a newly identified enteric alphacoronavirus that causes fatal diarrhea in newborn piglets in China. Here, we propagated a virulent strain SADS-CoV/CN/GDWT/2017 in Vero cells for up to 83 passages. Four strains of SADS-CoV/GDWT-P7, -P18, -P48 and -P83 were isolated and characterized. Sequence alignments showed that these four novel strains exhibited 16 nucleotide mutations and resultant 10 amino acid substitutions in open reading frame 1a/1b, spike, NS3a, envelope, membrane and nucleocapsid proteins. Furthermore, a 58-bp deletion in NS7a/7b was found in P48 and P83 strains, which led to the loss of NS7b and 38 amino acid changes of NS7a. Pig infection studies showed that the P7 strain caused typical watery diarrhea, while the P83 strain induced none-to-mild, delayed and transient diarrhea. This is the first report on cell adaption of a virulent SADS-CoV strain, which gives information on the potential virulence determinants of SADS-CoV.


title: Middle East respiratory syndrome: A new global threat

text: The outbreak of Middle East respiratory syndrome (MERS) is reported from Saudi Arabia and the Republic of Korea. It is a respiratory disease caused by coronavirus. Camels are considered as a source for MERS transmission in humans, although the exact source is unknown. Human-to-human transmission is reported in the community with droplet and contact spread being the possible modes. Most patients without any underlying diseases remain asymptomatic or develop mild clinical disease, but some patients require critical care for mechanical ventilation, dialysis and other organ support. MERS is a disease with pandemic potential and awareness, and surveillance can prevent such further outbreaks.


Online Balint groups in healthcare workers caring for the COVID-19 patients


COVID-19 and burns: lessons learned?


title: Understanding Transmission Dynamics of COVID-19-Type Infections by Direct Numerical Simulations of Cough/Sneeze Flows

text: The transmission dynamics of highly contagious respiratory diseases like COVID-19 (through coughing/sneezing) is an open problem in the epidemiological studies of such diseases (Bourouiba, JAMA. https://doi.org/10.1001/jama.2020.4756. 2020). The problem is basically the fluid dynamics of a transient turbulent jet/puff with buoyancy, laden with evaporating droplets carrying the pathogen. A turbulent flow of this nature does not lend itself to reliable estimates through modeling approaches such as RANS (Reynolds-Averaged Navier–Stokes equations) or other droplet-based models. However, direct numerical simulations (DNS) of what may be called “cough/sneeze flows” can play an important role in understanding the spread of the contagion. The objective of this work is to develop a DNS code for studying cough/sneeze flows by a suitable combination of the DNS codes available with the authors (developed to study cumulus cloud flows including thermodynamics of phase change and the dynamics of small water droplets) and to generate useful data on these flows. Recent results from the cumulus cloud simulations are included to highlight the effect of turbulent entrainment (which is one of the key processes in determining the spread of the expiratory flows) on the distribution of liquid water content in a moist plume. Furthermore, preliminary results on the temperature distribution in a “dry cough” (i.e., without inclusion of liquid droplets) are reported to illustrate the large spatial extent and time duration over which the cough flow can persist after the coughing has stopped. We believe that simulations of this kind can help to devise more accurate guidelines for separation distances between neighbors in a group, design better masks, and minimize the spread of respiratory diseases of the COVID-19 type.


title: Predictive factors of Covid-19 in patients with negative RT-qPCR

text: Abstract Objective: To evaluate the factors associated with false negatives in RT-qPCR in patients with mild-moderate symptoms of COVID-19. Materials and methods: This was a cross-sectional study that used a random sample of non-hospitalized patients from the primary care management division of the Healthcare Area of Leon (58 RT-qPCR-positive cases and 52 RT-qPCR-negative cases). Information regarding symptoms was collected and all patients were simultaneously tested using two rapid diagnostic test - RDTs (Combined - cRDT and Differentiated - dRDT). The association between symptoms and SARS-CoV-2 infection was evaluated by non-conditional logistic regression, with estimation of Odds Ratio. Results: A total of 110 subjects were studied, 52% of whom were women (mean age: 48.2 ± 11.0 years). 42.3% of negative RT-qPCRs were positive in some RDTs. Fever over 38ºC (present in 35.5% of cases) and anosmia (present in 41.8%) were the symptoms most associated with SARS-CoV-2 infection, a relationship that remained statistically significant in patients with negative RT-qPCR and some positive RDT (aOR=6.64; 95%CI=1.33-33.13 and aOR=19.38; 95% CI=3.69-101.89 respectively Conclusions: RT-qPCR is the technique of choice in the diagnosis of SARS-CoV-2 infection, but it is not exempt from false negatives. Our results show that those patients who present mild or moderate symptoms with negative RT-qPCR, but with fever and/or anosmia, should be considered suspicious cases and should be evaluated with other diagnostic methods.


title: Identifying Hosts of Families of Viruses: A Machine Learning Approach

text: Identifying emerging viral pathogens and characterizing their transmission is essential to developing effective public health measures in response to an epidemic. Phylogenetics, though currently the most popular tool used to characterize the likely host of a virus, can be ambiguous when studying species very distant to known species and when there is very little reliable sequence information available in the early stages of the outbreak of disease. Motivated by an existing framework for representing biological sequence information, we learn sparse, tree-structured models, built from decision rules based on subsequences, to predict viral hosts from protein sequence data using popular discriminative machine learning tools. Furthermore, the predictive motifs robustly selected by the learning algorithm are found to show strong host-specificity and occur in highly conserved regions of the viral proteome.


title: Perceived versus proven SARS-CoV-2-specific immune responses in health-care professionals

text: There have been concerns about high rates of thus far undiagnosed SARS-CoV-2 infections in the health-care system. The COVID-19 Contact (CoCo) Study follows 217 frontline health-care professionals at a university hospital with weekly SARS-CoV-2-specific serology (IgA/IgG). Study participants estimated their personal likelihood of having had a SARS-CoV-2 infection with a mean of 21% [median 15%, interquartile range (IQR) 5–30%]. In contrast, anti-SARS-CoV-2 IgG prevalence was about 1–2% at baseline. Regular anti-SARS-CoV-2 IgG testing of health-care professionals may aid in directing resources for protective measures and care of COVID-19 patients in the long run.


title: Infecciones transmitidas por el donante

text: El tratamiento inmunosupresor que recibe el receptor de un trasplante de órgano sólido dificulta la respuesta defensiva frente a la infección. La transmisión de la misma desde un donante puede provocar la disfunción o pérdida del injerto e, incluso, la muerte del receptor si no se establecen las medidas preventivas oportunas. Este riesgo potencial debe ser evaluado minuciosamente con la intención de optimizar el uso de órganos, especialmente en aquellos casos procedentes de donantes infectados, sin aumentar la disfunción del injerto y la morbimortalidad en el receptor. Este artículo pretende revisar los conocimientos actuales sobre el cribado de las infecciones en los donantes potenciales y discutir la relaciónriesgo-beneficio para usar órganos de donantes infectados. The defence response to infectious agents is compromised in solid organ recipients because of their immunosuppressive treatment. Transmission of infection from a donor organ can result in dysfunction or loss of the allograft, and may lead to death of the recipient if adequate preventive measures are not taken. This potential risk should be thoroughly assessed, particularly in the case of organs from infected donors, in order to optimize organ use without increasing the incidence of graft dysfunction and recipient morbidity and mortality. This article reviews the current knowledge regarding screening for infection in potential donors and discusses risk-benefit considerations related to the use of organs from infected donors.


title: A Study on the Contaminant Dispersion from Isolation Ward under Abnormal Operation of Facilities

text: Abstract Due to the recent outbreaks of infectious diseases, such as severe acute respiratory syndrome (SARS), Influenza and Ebola, isolation facilities have played an important role to prevent infectious diseases from spreading at initial stage. An isolation ward is a facility to isolate patients physically and to care them safely. One way to isolate a patient physically is to build a negative pressure isolation facility. However, unexpected failure or misuse of such facility makes it difficult to maintain negative pressure and eventually causes secondary infection, leaking the infectious pathogen to outside of the isolation ward. This study identifies the amount and velocity of leakage air from a patient ward by tracer gas experiment under abnormal operations of an isolation facility. In addition, computational fluid dynamics (CFD) allowed us to observe the outflow mechanism of pollutant. The results show that abnormal operations of a facility spreads pathogens to neighboring areas immediately and timely actions should be prepared against them.


title: Epidemics and food systems: what gets framed, gets done

text: This brief article aims to interrogate some widely used concepts in framing the interactions between disease epidemics, food systems and nutrition, with a particular focus on the COVID-19 crisis. How should we conceptualize vulnerability in such situations – both with regard to viral exposure and to subsequent nutrition-relevant impacts of epidemics and responses (including lockdowns)? Is it possible to simultaneously pursue strategies aimed at strengthening resilience and driving transformation (‘building back better’)? What type of framing and conceptualization can help illuminate entry points and options for responding effectively to interacting crises? In addressing these questions, it’s important to re-visit lessons from past attempts to address the impacts of epidemics on food and nutrition security.


title: LTB4 increases nasal neutrophil activity and conditions neutrophils to exert antiviral effects

text: Summary Background Leukotriene B4 (LTB4) recruits and activates neutrophils. Accordingly, this leukotriene is involved in innate defense actions. Objective To examine if nasal LTB4 can produce neutrophil activity and to explore whether or not LTB4 can condition neutrophils to exert virucidal effects in vitro and in vivo. Methods 1. Twenty-three healthy subjects received nasal LTB4 in a randomized and sham-controlled design. Symptoms were scored and nasal lavages carried out. Myeloperoxidase (MPO) and α-defensins were monitored as indices of neutrophil activity. IL-8, eosinophil cationic protein (ECP) and α2-macroglobulin were measured as indices of pro-inflammatory cytokine production, eosinophil activity, and plasma exudation. 2. Supernatants from neutrophils activated by LTB4 in vitro were assayed for virucidal activity against respiratory viruses. 3. In 38 healthy individuals, nasal inoculation with human rhinovirus-16 (HRV-16) was performed. In a preliminary study, intervention with LTB4 was given in a randomized and controlled design. Symptoms, virus replication, and antibody-titres were monitored. Results 1. LTB4 produced statistically significant increases in MPO and α-defensins, whereas IL-8, ECP, and α2-macroglobulin were unaffected. 2. The supernatants efficiently killed human coronavirus, respiratory syncytial virus, and influenza B virus. 3. HRV-16 replication was lower in subjects receiving LTB4, but this difference failed to reach statistical significance. Common cold symptoms and incidence of seroconversion were unaffected. Conclusion Nasal LTB4 induces a selective recruitment/activation of neutrophils. LTB4 can condition neutrophils to exert virucidal effects in vitro and may reduce virus replication in vivo. We suggest that the condition induced by LTB4 reflects an enhanced state of innate defense.


title: Uncertainty Shocks and Business Cycle Research

text: We review the literature on uncertainty shocks and business cycle research. First, we motivate the study of uncertainty shocks by documenting the presence of time-variation in the volatility of macroeconomic time series. Second, we enumerate the mechanisms that researchers have postulated to link uncertainty shocks and business cycles. Third, we outline how we can specify uncertainty shocks. Fourth, we postulate a real business cycle model augmented with financial frictions and uncertainty shocks. Fifth, we use the model to illustrate our previous discussions and to show how uncertainty shocks can be expansionary, a useful finding in several contexts.


title: Current Research on Non-Coding Ribonucleic Acid (RNA)

text: Non-coding ribonucleic acid (RNA) has without a doubt captured the interest of biomedical researchers. The ability to screen the entire human genome with high-throughput sequencing technology has greatly enhanced the identification, annotation and prediction of the functionality of non-coding RNAs. In this review, we discuss the current landscape of non-coding RNA research and quantitative analysis. Non-coding RNA will be categorized into two major groups by size: long non-coding RNAs and small RNAs. In long non-coding RNA, we discuss regular long non-coding RNA, pseudogenes and circular RNA. In small RNA, we discuss miRNA, transfer RNA, piwi-interacting RNA, small nucleolar RNA, small nuclear RNA, Y RNA, single recognition particle RNA, and 7SK RNA. We elaborate on the origin, detection method, and potential association with disease, putative functional mechanisms, and public resources for these non-coding RNAs. We aim to provide readers with a complete overview of non-coding RNAs and incite additional interest in non-coding RNA research.


title: SARS‐CoV‐2 and SARS‐CoV: Virtual screening of potential inhibitors targeting RNA‐dependent RNA polymerase activity (NSP12)

text: Since the outbreak of severe acute respiratory syndrome (SARS) in 2003, the harm caused by coronaviruses to the world cannot be underestimated. Recently, a novel coronavirus (severe acute respiratory syndrome coronavirus‐2 [SARS‐CoV‐2]) initially found to trigger human severe respiratory illness in Wuhan City of China in 2019, has infected more than six million people worldwide by 21 June 2020, and which has been recognized as a public health emergency of international concern as well. And the virus has spread to more than 200 countries around the world. However, the effective drug has not yet been officially licensed or approved to treat SARS‐Cov‐2 and SARS‐Cov infection. NSP12‐NSP7‐NSP8 complex of SARS‐CoV‐2 or SARS‐CoV, essential for viral replication and transcription, is generally regarded as a potential target to fight against the virus. According to the NSP12‐NSP7‐NSP8 complex (PDB ID: 7BW4) structure of SARS‐CoV‐2 and the NSP12‐NSP7‐NSP8 complex (PDB ID: 6NUR) structure of SARS‐CoV, NSP12‐NSP7 interface model, and NSP12‐NSP8 interface model were established for virtual screening in the present study. Eight compounds (Nilotinib, Saquinavir, Tipranavir, Lonafarnib, Tegobuvir, Olysio, Filibuvir, and Cepharanthine) were selected for binding free energy calculations based on virtual screening and docking scores. All eight compounds can combine well with NSP12‐NSP7‐NSP8 in the crystal structure, providing drug candidates for the treatment and prevention of coronavirus disease 2019 and SARS.


title: A method for the improved detection of aerosolized influenza viruses and the male-specific (F+) RNA coliphage MS2

text: Abstract The detection of aerosolized viruses can serve as an important surveillance and control tool in agriculture, human health, and environmental settings. Here, we adapted an anion exchange resin-based method, initially developed to concentrate negatively charged viruses from water, to liquid impingement-based bioaerosol sampling. In this method, aerosolized viruses are collected in a 20ml liquid sample contained within widely used impingers, BioSamplers (SKC Inc., Eighty Four, PA), and further concentrated via adsorption to an anion exchange resin that is suspended within this liquid. Viral nucleic acids are then extracted from the resin to facilitate molecular analyses through a reduction in the effective sample volume. For this study, various quantities of two negatively charged viruses, type A and type B influenza viruses (FluMist Quadrivalent vaccine) and the male-specific (F+) RNA coliphage MS2 (MS2), were nebulized into a custom-built bioaerosolization chamber, and sampled using BioSamplers with and without anion exchange resin. Compared to direct testing of the BioSampler liquid, detection was improved by 6.77× and 3.33× for type A and type B influenza viruses, respectively, by using the anion exchange resin. For MS2, the anion exchange resin method allowed for an average improvement in detection of 8.26×.


title: Convalescent plasma in Covid-19: Possible mechanisms of action

text: Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible of the coronavirus disease 2019 (COVID-19) pandemic. Therapeutic options including antimalarials, antivirals, antibiotics and vaccines are under study. Meanwhile the current pandemic has called attention over old therapeutic tools to treat infectious diseases. Convalescent plasma (CP) constitutes the first option in the current situation, since it has been successfully used in other coronaviruses outbreaks. Herein, we discuss the possible mechanisms of action of CP and their repercussion in COVID-19 pathogenesis, including direct neutralization of the virus, control of an overactive immune system (i.e., cytokine storm, Th1/Th17 ratio, complement activation) and immunomodulation of a hypercoagulable state. All these benefits of PC are expected to be better achieved if used in non-critically hospitalized patients, in the hope of reducing morbidity and mortality.


title: What did we learn from the previous coronavirus epidemics and what can we do better: a neuroinfectiological point of view

text: We are learning gradually about the neurologic manifestations during the ongoing coronavirus disease 2019 (COVID)‐19 pandemic, a respiratory disease related to severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2). As the clinical evidence is still limited, it makes a sense to critically analyze the spectrum of neurologic involvement caused by SARS‐CoV‐1 and Middle East respiratory syndrome coronavirus (MERS‐CoV), which were causal for epidemics in 2003 and 2012, respectively.


title: Evaluation of metal-conjugated compounds as inhibitors of 3CL protease of SARS-CoV

text: 3C-like (3CL) protease is essential for the life cycle of severe acute respiratory syndrome-coronavirus (SARS-CoV) and therefore represents a key anti-viral target. A compound library consisting of 960 commercially available drugs and biologically active substances was screened for inhibition of SARS-CoV 3CL protease. Potent inhibition was achieved using the mercury-containing compounds thimerosal and phenylmercuric acetate, as well as hexachlorophene. As well, 1–10 μM of each compound inhibited viral replication in Vero E6 cell culture. Detailed mechanism studies using a fluorescence-based protease assay demonstrated that the three compounds acted as competitive inhibitors (K(i)=0.7, 2.4, and 13.7 μM for phenylmercuric acetate, thimerosal, and hexachlorophene, respectively). A panel of metal ions including Zn(2+) and its conjugates were then evaluated for their anti-3CL protease activities. Inhibition was more pronounced using a zinc-conjugated compound (1-hydroxypyridine-2-thione zinc; Ki=0.17 μM) than using the ion alone (Ki=1.1 μM).


title: DC-SIGN and DC-SIGNR genetic diversity among different ethnic populations: Potential implications for pathogen recognition and disease susceptibility

text: Dendritic cell–specific intracellular adhesion molecule-3–grabbing nonintegrin (DC-SIGN) and DC-SIGNR are C-type lectins that serve both as cell adhesion and pathogen recognition receptors. Because of the essential role of the these molecules in the immune response, the implication of their alleles in human disease states, and the possible genetic variation at these loci among ethnically diverse populations, we undertook a study to analyze the full extent of DC-SIGN and DC-SIGNR polymorphisms in Caucasian Canadian and indigenous African populations. We report several novel nucleotide variants within regulatory 5′- and 3′-untranslated regions of the genes that could affect their transcription and translation. There were significant differences in the distribution of DC-SIGN and DC-SIGNR alleles among African and non-African populations. Finally, our study clearly demonstrates that Africans show greater genetic diversity at these two closely-related immune loci than observed in other major population groups. The differences may reflect evolutionary pressures generated by environmental factors, such as prevalent pathogens in these geographically distinct regions. Further studies will be needed to determine the net impact of DC-SIGN and DC-SIGNR genetic variants on the expression, translation, and function of the proteins and to understand how these functional polymorphisms may affect immune responses or immune escape.


title: NSAIDs in patients with viral infections, including Covid-19: victims or perpetrators?

text: Abstract Taking anti-inflammatory drugs, including non-steroidal (NSAIDs), during Covid-19 infection, how much is risky? The French Minister of Health, who has raised an alarm on a possible risk deriving from the use of ibuprofen for the control of fever and other symptoms during the disease, opened the debate a few days ago. In this paper we examine available evidence from preclinical and clinical studies that had analysed the role of COX in the inflammatory process and the effects of NSAIDs in patients with infections. Most of the published studies that suggested not protective effects of NSAIDs were mainly performed in vitro or on animals. Therefore, their meaning in humans is to be considered with great caution. Based also on data suggesting protective effects of NSAIDs, we concluded that currently there is no evidence suggesting a correlation between NSAIDs and a worsening of infections. Further studies will be certainly needed to better define the role of NSAIDs and particularly COX2 inhibitors in patients with infections. In the meantime, we must wait for results of the revision started by the PRAC on May 2019 on the association ibuprofen/ketoprofen and worsening of infections. Since nowadays no scientific evidence establishes a correlation between NSAIDS and worsening of COVID-19, patients should be advice against any NSAIDs self-medication when COVID-19 like symptoms are present.


title: Subgenomic messenger RNAs: Mastering regulation of (+)-strand RNA virus life cycle

text: Abstract Many (+)-strand RNA viruses use subgenomic (SG) RNAs as messengers for protein expression, or to regulate their viral life cycle. Three different mechanisms have been described for the synthesis of SG RNAs. The first mechanism involves internal initiation on a (−)-strand RNA template and requires an internal SGP promoter. The second mechanism makes a prematurely terminated (−)-strand RNA which is used as template to make the SG RNA. The third mechanism uses discontinuous RNA synthesis while making the (−)-strand RNA templates. Most SG RNAs are translated into structural proteins or proteins related to pathogenesis: however other SG RNAs regulate the transition between translation and replication, function as riboregulators of replication or translation, or support RNA–RNA recombination. In this review we discuss these functions of SG RNAs and how they influence viral replication, translation and recombination.


Let Us Aggressively Confront the Coronavirus Pandemic


title: Purification of Houttuynia cordata Thunb. Essential Oil Using Macroporous Resin Followed by Microemulsion Encapsulation to Improve Its Safety and Antiviral Activity

text: Essential oil extracted from Houttuynia cordata Thunb. (H. cordata) is widely used in traditional Chinese medicine due to its excellent biological activities. However, impurities and deficient preparations of the essential oil limit its safety and effectiveness. Herein, we proposed a strategy to prepare H. cordata essential oil (HEO) safely and effectively by combining the solvent extraction and the macroporous resin purification flexibly, and then encapsulating it using microemulsion. The extraction and purification process were optimized by orthogonal experimental design and adsorption-desorption tests, respectively. The average houttuynin content in pure HEO was then validated at 44.3% ± 2.01%, which presented a great potential for industrial application. Subsequently, pure HEO-loaded microemulsion was prepared by high-pressure homogenization and was then fully characterized. Results showed that the pure HEO-loaded microemulsion was successfully prepared with an average particle size of 179.1 nm and a high encapsulation rate of 94.7%. Furthermore, safety evaluation tests and in vitro antiviral testing indicated that the safety and activity of HEO were significantly improved after purification using D101 resin and were further improved by microemulsion encapsulation. These results demonstrated that the purification of HEO by macroporous resin followed by microemulsion encapsulation would be a promising approach for industrial application of HEO for the antiviral therapies.


title: A comparison of three rapid diagnostic methods for the detection of rotavirus infection in calves

text: Abstract Three techniques for the detection of rotavirus in faecal samples from calves with neonatal gastroenteritis were compared. A preliminary study indicated that reverse passive haemagglutination (RPHA) was at least as sensitive as the enzyme-linked immunosorbent assay (ELISA). These two immunoassays were compared with the detection of viral RNA by polyacrylamide gel electrophoresis (PAGE) on 209 field samples. Of the 77 samples in which at least one test gave a positive result, 69 were positive by both RPHA and PAGE, but only 49 were also positive by ELISA, indicating a lower sensitivity for the latter test. The overall agreement between RPHA and PAGE was 96%. The reasons for the discrepancies between the tests are discussed.


title: Expression of the SARS-CoV-2 cell receptor gene ACE2 in a wide variety of human tissues

text: BACKGROUND: Since its discovery in December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 2 180 000 people worldwide and has caused more than 150 000 deaths as of April 16, 2020. SARS-CoV-2, which is the virus causing coronavirus disease 2019 (COVID-19), uses the angiotensin-converting enzyme 2 (ACE2) as a cell receptor to invade human cells. Thus, ACE2 is the key to understanding the mechanism of SARS-CoV-2 infection. This study is to investigate the ACE2 expression in various human tissues in order to provide insights into the mechanism of SARS-CoV-2 infection. METHODS: We compared ACE2 expression levels across 31 normal human tissues between males and females and between younger (ages ≤ 49 years) and older (ages > 49 years) persons using two-sided Student’s t test. We also investigated the correlations between ACE2 expression and immune signatures in various tissues using Pearson’s correlation test. RESULTS: ACE2 expression levels were the highest in the small intestine, testis, kidneys, heart, thyroid, and adipose tissue, and were the lowest in the blood, spleen, bone marrow, brain, blood vessels, and muscle. ACE2 showed medium expression levels in the lungs, colon, liver, bladder, and adrenal gland. ACE2 was not differentially expressed between males and females or between younger and older persons in any tissue. In the skin, digestive system, brain, and blood vessels, ACE2 expression levels were positively associated with immune signatures in both males and females. In the thyroid and lungs, ACE2 expression levels were positively and negatively associated with immune signatures in males and females, respectively, and in the lungs they had a positive and a negative correlation in the older and younger groups, respectively. CONCLUSIONS: Our data indicate that SARS-CoV-2 may infect other tissues aside from the lungs and infect persons with different sexes, ages, and races equally. The different host immune responses to SARS-CoV-2 infection may partially explain why males and females, young and old persons infected with this virus have markedly distinct disease severity. This study provides new insights into the role of ACE2 in the SARS-CoV-2 pandemic.


Hygiène des mains et masques contre COVID-19 : oser le dégradé ?


Sharing patient-level real-time COVID-19 data


title: Coronavirus Disease 2019, School Closures, and Children’s Mental Health

text: Coronavirus disease 2019 (COVID-19), which was first identified in Wuhan, China, in late December of 2019 is rapidly spreading across the globe. The South Korean government has ordered the closure of all schools, as part of its attempts to use social distancing measures to prevent the spread of COVID-19. The effects of the school closures on reducing contagion are generally positive; however, the measure is controversial because of the socioeconomic ripple effect that accompanies it. The author briefly reviewed the existing literature on the mental health aspects of disasters and presents the issues related to school closures due to pandemics, from medical and socioeconomic perspectives and in terms of children’s mental health. The results of this review suggest that research on children’s mental health in relation to the adoption of school closures as a pandemic mitigation strategy is urgently needed.


title: Approach to Common Bacterial Infections Community-Acquired Pneumonia

text: Community-acquired pneumonia (CAP) occurs more often in early childhood than at almost any other age. Many microorganisms are associated with pneumonia, but individual pathogens are difficult to identify, which poses problems in antibiotic management. This article reviews the common as well as new, emerging pathogens, as well as the guidelines for management of pediatric CAP. Current guidelines for pediatric CAP continue to recommend the use of high-dose amoxicillin for bacterial CAP and azithromycin for suspected atypical CAP (usually caused by Mycoplasma pneumoniae) in children.


title: Passive protection effect of chicken egg yolk immunoglobulins on enterovirus 71 infected mice

text: The objective of this study is to evaluate the passive protective efficiency of immunoglobulin in yolk (IgY) specific against human enterovirus type 71 (EV71). The antibody was raised by intramuscular immunization to 10 White Leghorn hens, with inactivated human EV71 serving as the antigen. The titer and specificity of the antibody were analyzed from purified IgY in the egg yolks of immunized hens. Results indicate that the titer of IgY specific against EV71 increased from the third week after the first immunization. The content of total IgY was 190 ± 26 mg/yolk, with an average concentration of specific IgY of 6.34 ± 3.38 mg/yolk in the eggs from 3 to 18 wk after immunization. The results of the neutralization effect of specific IgY in EV71-challenged mice demonstrate that the EV71-specific IgY, either by intraperitoneal injection or oral administration, was able to significantly reduce the morbidity and mortality in EV71 infected mice pups.


title: Comparison of three diagnostic techniques for detection of rotavirus and coronavirus in calf faeces in Australia

text: Objective Compare real‐time reverse transcription polymerase chain reaction (qRT‐PCR), a commercially available enzyme‐linked immunosorbent assay (ELISA) and lateral flow immunochromatography assay (LAT) for the detection of rotavirus and coronavirus in faecal samples collected from diarrhoeic calves. Design Prospective survey. Method Samples were tested at two separate facilities using a commercial ELISA and an in‐house qRT‐PCR. Simple logistic regression was performed to examine the relationship between the two tests. A subset of samples was screened using qRT‐PCR, ELISA and a commercial LAT dipstick (132 faecal samples were tested for coronavirus and 122 samples for rotavirus). Results Of the 586 samples tested, 131 (22.39%) and 468 (79.86%) were positive for coronavirus and group A rotavirus, respectively, using qRT‐PCR. The number of samples positive on ELISA for coronavirus and rotavirus was 73 (12.46%) and 225 (38.40%), respectively. Using LAT, 30 (22.73%) and 43 (35.35%) samples were positive for coronavirus and rotavirus, respectively. Simple linear regression revealed a statistically significant (P < 0.05) but weak (r(2)=−0.07 and −0.40) correlation between the rotavirus/coronavirus qRT‐PCR and ELISA, respectively. There was also poor agreement between the LAT and qRT‐PCR assays. Conclusion The sensitivity and specificity of the commercial ELISA and LAT assays evaluated in this study were low compared with qRT‐PCR. The low positive and negative predictive values of the assays suggests that they were of limited diagnostic benefit in the population sampled.


title: Comprehensive Molecular Testing for Respiratory Pathogens in Community-Acquired Pneumonia

text: Background. The frequent lack of a microbiological diagnosis in community-acquired pneumonia (CAP) impairs pathogen-directed antimicrobial therapy. This study assessed the use of comprehensive multibacterial, multiviral molecular testing, including quantification, in adults hospitalized with CAP. Methods. Clinical and laboratory data were collected for 323 adults with radiologically-confirmed CAP admitted to 2 UK tertiary care hospitals. Sputum (96%) or endotracheal aspirate (4%) specimens were cultured as per routine practice and also tested with fast multiplex real-time polymerase-chain reaction (PCR) assays for 26 respiratory bacteria and viruses. Bacterial loads were also calculated for 8 bacterial pathogens. Appropriate pathogen-directed therapy was retrospectively assessed using national guidelines adapted for local antimicrobial susceptibility patterns. Results. Comprehensive molecular testing of single lower respiratory tract (LRT) specimens achieved pathogen detection in 87% of CAP patients compared with 39% with culture-based methods. Haemophilus influenzae and Streptococcus pneumoniae were the main agents detected, along with a wide variety of typical and atypical pathogens. Viruses were present in 30% of cases; 82% of these were codetections with bacteria. Most (85%) patients had received antimicrobials in the 72 hours before admission. Of these, 78% had a bacterial pathogen detected by PCR but only 32% were culture-positive (P < .0001). Molecular testing had the potential to enable de-escalation in number and/or spectrum of antimicrobials in 77% of patients. Conclusions. Comprehensive molecular testing significantly improves pathogen detection in CAP, particularly in antimicrobial-exposed patients, and requires only a single LRT specimen. It also has the potential to enable early de-escalation from broad-spectrum empirical antimicrobials to pathogen-directed therapy.


title: Financial volatility trading using a self-organising neural-fuzzy semantic network and option straddle-based approach

text: Abstract Financial volatility refers to the intensity of the fluctuations in the expected return on an investment or the pricing of a financial asset due to market uncertainties. Hence, volatility modeling and forecasting is imperative to financial market investors, as such projections allow the investors to adjust their trading strategies in anticipation of the impending financial market movements. Following this, financial volatility trading is the capitalization of the uncertainties of the financial markets to realize investment profits in times of rising, falling and side-way market conditions. In this paper, an intelligent straddle trading system (framework) that consists of a volatility projection module (VPM) and a trade decision module (TDM) is proposed for financial volatility trading via the buying and selling of option straddles to help a human trader capitalizes on the underlying uncertainties of the Hong Kong stock market. Three different measures, namely: (1) the historical volatility (HV), (2) implied volatility (IV) and (3) model-based volatility (MV) of the Hang Seng Index (HSI) are employed to quantify the implicit volatility of the Hong Kong stock market. The TDM of the proposed straddle trading system combines the respective volatility measures with the well-established moving-averages convergence/divergence (MACD) principle to recommend trading actions to a human trader dealing in HSI straddles. However, the inherent limitation of the MACD trading rule is that it generates time-delayed trading signals due to the use of moving averages, which are essentially lagging trend indicators. This drawback is intuitively addressed in the proposed straddle trading system by applying the VPM to compute future projections of the volatility measures of the HSI prior to the activation of the TDM. The VPM is realized by a self-organising neural-fuzzy semantic network named the evolving fuzzy semantic memory (eFSM) model. As compared to existing statistical and computational intelligence based modeling techniques currently employed for financial volatility modeling and forecasting, eFSM possesses several desirable attributes such as: (1) an evolvable knowledge base to continuously address the non-stationary characteristics of the Hong Kong stock market; (2) highly formalized human-like information computations; and (3) a transparent structure that can be interpreted via a set of linguistic IF–THEN semantic fuzzy rules. These qualities provide added credence to the computed HSI volatility projections. The volatility modeling and forecasting performances of the eFSM, when benchmarked to several established modeling techniques, as well as the observed trading returns of the proposed straddle trading system, are encouraging.


title: Peptide Mimicrying Between SARS Coronavirus Spike Protein and Human Proteins Reacts with SARS Patient Serum

text: Molecular mimicry, defined as similar structures shared by molecules from dissimilar genes or proteins, is a general strategy used by pathogens to infect host cells. Severe acute respiratory syndrome (SARS) is a new human respiratory infectious disease caused by SARS coronavirus (SARS-CoV). The spike (S) protein of SARS-CoV plays an important role in the virus entry into a cell. In this study, eleven synthetic peptides from the S protein were selected based on its sequence homology with human proteins. Two of the peptides D07 (residues 927–937) and D08 (residues 942–951) were recognized by the sera of SARS patients. Murine hyperimmune sera against these peptides bound to proteins of human lung epithelial cells A549. Another peptide D10 (residues 490–502) stimulated A549 to proliferate and secrete IL-8. The present results suggest that the selected S protein regions, which share sequence homology with human proteins, may play important roles in SARS-CoV infection.


title: EPIDEMIOLOGICAL AND CLINICAL CHARACTERISTICS OF HEART TRANSPLANT RECIPIENTS DURING THE 2019 CORONAVIRUS OUTBREAK IN WUHAN, CHINA: A DESCRIPTIVE SURVEY REPORT

text: ABSTRACT BACKGROUND The epidemiological and clinical characteristics of heart transplant (HTx) recipients during the SARS-CoV-2 epidemic remains unclear. We studied the characteristics of HTx recipients during Dec 20, 2019 and Feb 25, 2020 in an effort to understand their risk and outcomes. METHODS All accessible HTx recipients were included in this single-center, retrospective study. We collected information on the recipients using a web-based questionnaire as well as the hospital database. RESULTS We followed 87 HTx recipients (72.4% were men and average age was 51 years). 79 recipients resided in Hubei, and 57 recipients had a Wuhan related history of travel or contact. The majority took precautionary measures while in contact with suspicious crowds. 96.6% of the families and communities undertook prevention and quarantine procedures. 4 upper airway infection were reported, and 3 of them tested negative for SARS-CoV-2 (the 4th recovered and was not tested). All cases were mild and successfully recovered after proper treatment. Laboratory results of 47 HTx cases within the last 2 months were extracted. 21.3% recipients had pre-existing lymphopenia and 87.2% of recipients had therapeutic tacrolimus concentration (5-12ng/ml). 5 and 6 recipients had liver and kidney insufficiency respectively. CONCLUSION HTx recipients that practiced appropriate prevention measures had a low rate of infection with SARS-CoV-2 and transition to COVID-19. These early data will require confirmation as the pandemic establishes around the world.


A planetary vision for one health


title: No short-term effect of handling and capture stress on immune responses of bats assessed by bacterial killing assay

text: Abstract Ecoimmunology of wild animals becomes increasingly important. However, there are methodical limitations, especially when working on small mammals, e.g. small sample volume and acute stress associated with capture, handling and sampling that can influence immune parameters. The plasma bacterial killing assay measures innate humoral immune responses, mainly complement activity. It is a powerful tool with many methodical advantages. To avoid investigation of artefacts in future ecoimmunological studies the influence of acute stress on the bacterial killing activity was assessed. Bats (Nyctalus noctula, n =9) were repeatedly sampled in three time intervals up to 97min after capture. Bacterial killing activity against Escherichia coli was measured using a microplate absorbance reader. Bacterial killing activity was not influenced by capture, handling and sampling. Hence, released stress hormones did not affect circulating complement activity. To conclude, the plasma bacterial killing assay is reliable and efficient ecoimmunological tool in wildlife studies even of small mammals.


title: Veterinary vaccine development from an industrial perspective

text: Abstract Veterinary vaccines currently available in Europe and in other parts of the world are developed by the veterinary pharmaceutical industry. The development of a vaccine for veterinary use is an economic endeavour that takes many years. There are many obstacles along the path to the successful development and launch of a vaccine. The industrial development of a vaccine for veterinary use usually starts after the proof of concept that is based on robust academic research. A vaccine can only be made available to the veterinary community once marketing authorisation has been granted by the veterinary authorities. This review gives a brief description of the regulatory requirements which have to be fulfilled before a vaccine can be admitted to the market. Vaccines have to be produced in a quality controlled environment to guarantee delivery of a product of consistent quality with well defined animal and consumer safety and efficacy characteristics. The regulatory and manufacturing legislative framework in which the development takes place is described, as well as the trend in developments in production systems. Recent developments in bacterial, viral and parasite vaccine research and development are also addressed and the development of novel adjuvants that use the expanding knowledge of immunology and disease pathology are described.


title: Clinical Characteristics, Management and Prognostic Factors in Patients with Probable Severe Acute Respiratory Syndrome (SARS) in a SARS Center in Taiwan

text: Background Severe acute respiratory syndrome (SARS) is an emerging viral infectious disease. We report our experience in treating SARS patients. Methods From April 27 to May 24, 2003, a total of 36 patients with probable SARS were admitted and treated in a hospital rearranged as a special center for the management of SARS patients. Medical records for the patients were retrospectively reviewed. Univariate and multivariate analyses were performed to determine factors associated with respiratory failure and intubation. Results Of the 36 patients with probable SARS (median age, 37 years; range, 22-66 years), 9 were male and 27 were female. Thirty-two patients (88.9%) were infected in the hospital setting. All patients presented with fever, and 33 eventually developed lymphopenia during hospitalization. Chest radiography showed no unique pattern, but pleural effusion was not seen. All patients initially received empiric antibacterial therapy against common causative pathogens of atypical pneumonia. Ribavirin was given to all except 1 patient. Twenty-two patients received immunoglobulin therapy, and 32 were given corticosteroids. A total of 20 patients (55.6%) required supplemental oxygen, and 8 (22.2%) were intubated with mechanical ventilatory support. Two of these patients died. A higher body temperature at presentation (median 39.5 vs 38.6°C), and higher peak values of lactate dehydrogenase (410 vs 282 U/L) and C-reactive protein (10.2 vs 2.5 mg/dL), were associated with subsequent respiratory failure. Multivariate analysis showed that peak level of C-reactive protein was the only independent predictor of respiratory failure and intubation (odds ratio for every increment of 1 mg/dL = 1.45; 95% confidence interval = 1.003, 2.097; p = 0.048). Conclusion All patients with probable SARS who were admitted to hospital presented with fever and lymphopenia. While the efficacy of different treatments could not be evaluated from this retrospective study, a higher value of C-reactive protein was associated with the development of respiratory failure and subsequent intubation.


title: Antibiotic Prescribing and Respiratory Viral Testing for Acute Upper Respiratory Infections Among Adult Patients at an Ambulatory Cancer Center

text: BACKGROUND: Outpatient antibiotic prescribing for acute upper respiratory infections (URIs) is a high-priority target for antimicrobial stewardship that has not been described for cancer patients. METHODS: We conducted a retrospective cohort study of adult patients at an ambulatory cancer center with URI diagnoses from 1 October 2015 to 30 September 2016. We obtained antimicrobial prescribing, respiratory viral testing, and other clinical data at first encounter for the URI through day 14. We used generalized estimating equations to test associations of baseline factors with antibiotic prescribing. RESULTS: Of 341 charts reviewed, 251 (74%) patients were eligible for analysis. Nearly one-third (32%) of patients were prescribed antibiotics for URIs. Respiratory viruses were detected among 85 (75%) of 113 patients tested. Antibiotic prescribing (P = .001) and viral testing (P < .001) varied by clinical service. Sputum production or chest congestion was associated with higher risk of antibiotic prescribing (relative risk [RR], 2.3; 95% confidence interval [CI], 1.4–3.8; P < .001). Viral testing on day 0 was associated with lower risk of antibiotic prescribing (RR, 0.4; 95% CI 0.2–0.8; P = .01), though collinearity between viral testing and clinical service limited our ability to separate these effects on prescribing. CONCLUSIONS: Nearly one-third of hematology–oncology outpatients were prescribed antibiotics for URIs, despite viral etiologies identified among 75% of those tested. Antibiotic prescribing was significantly lower among patients who received an initial respiratory viral test. The role of viral testing in antibiotic prescribing for URIs in outpatient oncology settings merits further study.


title: Reconstitution of bluetongue virus polymerase activity from isolated domains based on a three‐dimensional structural model

text: Bluetongue virus (BTV) is a double‐stranded RNA virus of the Reoviridae family. The VP1 protein of BTV is the viral RNA‐dependent RNA polymerase (RdRp), which is responsible for the replication of the viral genome. Currently there is no structural information available for VP1. By manual alignment of BTV, Reovirus and other viral RdRps we have generated a model for the structure of VP1, the RdRp of BTV. The structure can be divided into three domains: an N‐terminal domain, a C‐terminal domain, and a central polymerase domain. Mutation of the putative catalytic site in the central polymerase domain by site‐directed mutagenesis abrogated in vitro replicase activity. Each of the domains was expressed individually and subsequently partially purified to obtain direct evidence for the location of polymerase activity and the nucleoside triphosphate binding site. The nucleoside triphosphate binding site was located by showing that CTP only bound to the full‐length protein or to the polymerase domain and not to either of the other two domains. None of the domains had catalytic activity when tested individually or in tandem but when all three domains were mixed together the RdRp activity was reconstituted. This is the first report of the reconstitution of a functional viral RdRp in vitro from individual domains. © 2007 Wiley Periodicals, Inc. Biopolymers 86: 83–94, 2007. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com


title: Seroprevalence of porcine respiratory coronavirus in selected Korean pigs

text: Abstract A total of 446 serum samples from 88 herds in Korea were examined for antibody to porcine respiratory coronavirus (PRCV) using blocking enzyme-linked immunosorbent assay (ELISA). All serum samples were collected from 24- to 26-week-old finishing pigs between December 1998 and June 1999. By ELISA, 237 out of 446 sera tested (53.1%) and 54 out of 88 sampled herds (61.3%) were positive against PRCV. Of 446 sera from 88 herd tested, 185 (41.5%) serum samples from 22 (25%) herds were seronegative against PRCV and transmissible gastroenteritis virus infection. Our data suggested that seropositive herds for PRCV are distributed diffusely throughout South Korea.


title: Lessons learned and unlearned in periodontal microbiology

text: Periodontal diseases are initiated by bacterial species living in polymicrobial biofilms at or below the gingival margin and progress largely as a result of the inflammation initiated by specific subgingival species. In the past few decades, efforts to understand the microbiota of periodontal diseases have led to an exponential increase in information about biofilms associated with periodontal health and disease. In fact, the oral microbiota is one of the best characterized microbiomes that colonize the human body. Despite this increased knowledge, one has to ask if our fundamental concepts of the etiology and pathogenesis of periodontal diseases have really changed. In this chapter we will review how our comprehension of the structure and function of the subgingival microbiota evolved over the years in search of lessons learned and unlearned in periodontal microbiology. More specifically, this review focuses on: 1) how the data obtained through molecular techniques has impacted our knowledge of the etiology of periodontal infections; 2) the potential role of viruses in the etiopathogenesis of periodontal diseases; 3) how concepts of microbial ecology have expanded our understanding of host microbial interactions that might lead to periodontal diseases; 4) the role of inflammation in the pathogenesis of periodontal diseases; and 5) the impact of these evolving concepts on treatment and preventive approaches to periodontal infections. We will conclude by reviewing how novel systems biology approaches promise to unravel new details of the pathogenesis of periodontal diseases and, hopefully, lead to a better understanding of periodontal disease mechanisms.


title: Does weather affect the growth rate of COVID-19, a study to comprehend transmission dynamics on human health

text: Abstract The undefendable outbreak of novel coronavirus (SARS-COV-2) lead to a global health emergency due to its higher transmission rate and longer symptomatic duration, created a health surge in a short time. Since Nov 2019 the outbreak in China, the virus is spreading exponentially everywhere. The current study focuses on the relationship between environmental parameters and the growth rate of COVID-19. The statistical analysis suggests that the temperature changes retarded the growth rate and found that -6.28°C and +14.51°C temperature is the favorable range for COVID-19 growth. Gutenberg- Richter's relationship is used to estimate the mean daily rate of exceedance of confirmed cases concerning the change in temperature. Indeed, temperature is the most influential parameter that reduces the growth at the rate of 13-17 cases/day with a 1°C rise in temperature.


title: Typical pollutants in bottom ashes from a typical medical waste incinerator

text: Abstract Incineration of medical waste (MW) is an important alternative way for disposal of this type of hazardous waste, especially in China because of the outbreak of severe acute respiratory syndromes (SARs) in 2003. Thus, far, fly ash has received much attention but less attention has been paid to bottom ash. In this study, bottom ash samples were collected from a typical MW incinerator, and typical pollutants including heavy metals and polycyclic aromatic hydrocarbons (PAHs) in the ash were examined. X-ray fluorescence spectroscopy results indicated that CaO, SiO2 and Al2O3 were the main components of the bottom ash. Inductively coupled plasma-optical emission spectroscopy showed that the ash contained large amounts of heavy metals, including Zn, Ti, Ba, Cu, Pb, Mn, Cr, Ni and Sn. Most of the heavy metals (e.g., Ba, Cr, Ni, and Sn) presented in the residual fraction; whereas Mn, Pb and Zn presented in Fe–Mn oxides fraction, and Cu in organic-matter fraction. Toxicity characteristic leaching procedure tests indicated that the leached amounts of heavy metals were well below the limits. The sum of 16 US EPA priority PAHs (ΣPAHs) varied from 10.30 to 38.14mgkg−1, and the total amounts of carcinogenic PAHs ranged between 4.09 and 16.95mgkg−1, exceeding the limits regulated by several countries. This research provides basic information for the evaluation of the environmental risk of MW incinerator bottom ash.


title: ATP1B3: a virus-induced host factor against EV71 replication by up-regulating the production of type-I interferons

text: Abstract Enterovirus 71 (EV71) infection can cause severe diseases, and is becoming increasingly common in children. In the current study, we carried out yeast two-hybrid assays to screen human proteins that could interact with 3A protein of EV71. Human β3 subunit of Na+/K+-ATPase (ATP1B3) protein was demonstrated to interact with the 3A protein of EV71. Although 3A protein had no effect on the expression of ATP1B3, EV71 infection resulted in elevated expression of ATP1B3 in RD cell line, both on messenger RNA (mRNA) and protein levels. Interestingly, knockdown of ATP1B3 could significantly increase the replication of EV71, whereas overexpression of ATP1B3 significantly suppressed the replication of EV71 in RD cells. Furthermore, we demonstrated that the expression of ATP1B3 could induce the production of type-I interferons. Our study demonstrated that ATP1B3 inhibit EV71 replication by enhancing the production of type-I interferons, which could act as a potential therapeutic target in EV71 infection.


Reply to: “Skin damage among health care workers managing coronavirus disease-2019”


title: Confronting the avian influenza threat: vaccine development for a potential pandemic

text: Summary Sporadic human infection with avian influenza viruses has raised concern that reassortment between human and avian subtypes could generate viruses of pandemic potential. Vaccination is the principal means to combat the impact of influenza. During an influenza pandemic the immune status of the population would differ from that which exists during interpandemic periods. An emerging pandemic virus will create a surge in worldwide vaccine demand and new approaches in immunisation strategies may be needed to ensure optimum protection of unprimed individuals when vaccine antigen may be limited. The manufacture of vaccines from pathogenic avian influenza viruses by traditional methods is not feasible for safety reasons as well as technical issues. Strategies adopted to overcome these issues include the use of reverse genetic systems to generate reassortant strains, the use of baculovirusexpressed haemagglutinin or related non-pathogenic avian influenza strains, and the use of adjuvants to enhance immunogenicity. In clinical trials, conventional surfaceantigen influenza virus vaccines produced from avian viruses have proved poorly immunogenic in immunologically naive populations. Adjuvanted or whole-virus preparations may improve immunogenicity and allow sparing of antigen.


title: Human Bocavirus in Iranian children with acute gastroenteritis

text: BACKGROUND: Human Bocavirus (HBoV) infection is of worldwide distribution. There is increasing evidencethat HBoV is pathogenic for the human gastroenteric tract. However, less data are available on the role of HBoVin gastroenteritis. The present study was aimed to determine the prevalence of HBoV in children with gastroenteritis. METHODS: Real-time PCR TaqMan was used to screen 200 stool specimens that had been referred to the virologylaboratory for HBoV evaluation. All of samples were collected on viral transport media. Results:Of the 200 stool samples analyzed, 16 (8%) were positive for HBoV. Human Bocavirus positive samplesfrom patients aged between 1 to 5 years with acute gastroenteritis infection suggest a minor role of HBoVin gastroenteritis (p=0.0001). CONCLUSION: The study showed a high prevalence of human Bocavirus in young children with acute gastroenteritisdiseases in Iran, suggesting that HBoV play a role in the pathogenesis of gastroenteritis.


Highlights in this issue.


title: Increased pulmonary secretion of tumor necrosis factor-α in calves experimentally infected with bovine respiratory syncytial virus

text: Abstract Bovine respiratory syncytial virus (BRSV) is an important cause of respiratory disease among calves in the Danish cattle industry. An experimental BRSV infection model was used to study the pathogenesis of the disease in calves. Broncho alveolar lung lavage (BAL) was performed on 28 Jersey calves, of which 23 were experimentally infected with BRSV and five were given a mock inoculum. The presence of the cytokine tumor necrosis factor α (TNF-α) in the BAL fluids was detected and quantified by a capture ELISA. TNF-α was detected in 21 of the infected animals. The amount of TNF-α in the BAL fluid of calves killed post inoculation day (PID) 2 and 4 was at the same very low level as in the uninfected control animals. Large amounts of TNF-α were detected on PID 6, maximum levels of TNF-α were reached on PID 7, and smaller amounts of TNF-α were seen on PID 8. The high levels of TNF-α appeared on the days where severe lung lesions and clinical signs were obvious and the amounts of BRSV-antigen were at their greatest. Although Pasteurellaceae were isolated from some of the BRSV-infected calves, calves treated with antibiotics before and through the whole period of the infection, as well as BRSV-infected calves free of bacteria reached the same level of TNF-α as animals from which bacteria were isolated from the lungs. It is concluded that significant quantities of TNF-α are produced in the lungs of the calves on PID 6–7 of BRSV infection. The involvement of TNF-α in the pathogenesis of, as well as the anti-viral immune response against, BRSV infection is discussed.


title: Management of hospitalized drug sensitive pulmonary tuberculosis patients during the Hajj mass gathering: A cross sectional study

text: Abstract Background To document the management of drug-sensitive TB patients during the Hajj and assess compliance with the Saudi TB management guidelines. Method The study was conducted in hospitals in Makkah during the 2016 and 2017 Hajj seasons. Structured questionnaire was used to collect data on relevant indices on TB management and a scoring system was developed to assess compliance with guidelines. Results Data was collected from 31 TB cases, 65.4% (17/26) were Saudi residents. Sputum culture was the only diagnostic test applied in 67.7% (21/31) of patients. Most (96.8%, 30/31) confirmed TB cases were isolated, but only 12.9% (4/28) were tested for HIV and merely 37% (10/27) received the recommended four 1st-line anti-TB drugs. Guideline compliance scores were highest for infection prevention and control and surveillance (9.6/10) and identifying TB suspects (7.2/10). The least scores were obtained for treating TB (5.0/10) and diagnosing TB (3.0/10). Conclusions Healthcare providers training and supervision are paramount to improve their knowledge and skill and ensure their compliance with existing TB management guidelines. However, there may be a need for the introduction of an international policy/guideline for TB control and management during mass gatherings such as the Hajj to guide providers’ choices and facilitate monitoring.


title: Is Increased Sleep Responsible for Reductions in Myocardial Infarction During the COVID-19 Pandemic?

text: The COVID-19 pandemic caused by the highly contagious SARS-CoV-2 virus has had devastating consequences across the globe. However, multiple clinics and hospitals have experienced a decrease in rates of acute myocardial infarction and corresponding cardiac catheterization lab activations, raising the question: Has the risk of myocardial infarction decreased during COVID? Sleep deprivation is known to be an independent risk factor for myocardial infarction, and sleep has been importantly impacted during the pandemic, possibly due to the changes in work-home life leading to a lack of structure. We conducted a social media-based survey to assess potential mechanisms underlying the observed improvement in risk of myocardial infarction. We used validated questionnaires to assess sleep patterns, tobacco consumption and other important health outcomes to test the hypothesis that increases in sleep duration may be occurring which have a beneficial impact on health. We found that the COVID-19 pandemic led to shifts in circadian rhythm, with subjects waking up 105 minutes later during the pandemic (p < 0.0001). Subjects also reported going to sleep 41 minutes later during the pandemic (p < 0.0001). These shifts led to longer duration of sleep during the COVID-19 pandemic. Before the pandemic, subjects reported sleeping 6.8 hours per night, which rose to 7.5 hours during the pandemic, a 44 minute or 11% increase (p < 0.0001). We acknowledge the major negative health impact of the global pandemic but would advocate for using this crisis to improve the work and sleep habits of the general population, which may lead to overall health benefits for our society.


title: Clinical Testing For Covid-19

text: Abstract As the novel coronavirus SARS-CoV-2 caused COVID-19 cases in the United States the initial test was developed and performed at the Center for Disease Control (CDC). As the number of cases increased the demand for tests multiplied, leading the CDC to utilize the Emergency Utilization Authorization to allow clinical and commercial laboratories to develop tests to detect the presence of the virus. Many nucleic acid tests based on reverse transcriptase-polymerase chain reaction (RT-PCR) were developed, each with different techniques, specifications and turnaround time. As the illnesses turned into a pandemic, testing became more crucial. The test supply became inadequate to meet the need that it had to be prioritized according to guidance. For surveillance, the need for serologic tests emerged. Here we review the timeline of test development, the turn-around times, the various approved tests and compare them as regards the genes they detect. We concentrate on the point-of-care tests and discuss the basis for new serologic tests. We discuss the testing guidance for prioritization and their application in a hospital setting. As SARS-CoV-2 virus arrived in the USA causing the COVID-19 illness, one of the most talked about issues in the management of the disease and the resulting pandemic has been clinical testing. A unique situation arose of a communicable and highly contagious disease necessitating the rapid diagnosis of patients and the identification of non-symptomatic infected persons. Unfortunately, the USA did not have a Food and Drug Administration (FDA) approved laboratory test for the illness. The FDA ultimately utilized its Emergency Use Authorizations (EUA) on February 4, 2020 to allow for more rapid and widespread development and implementation of in-vitro testing.1 Indeed, companies and organizations utilized the EUA to file applications for new tests based on different methodologies, amounting to 48 applications in the span of 3 months from the beginning of February to the end of April 2020. In addition, multiple other tests were put in place under a separate authorization by a Presidential memorandum in early March allowing laboratories that carry Clinical Laboratory Improvement Amendment (CLIA) certification to put tests in place without an EUA from the FDA. This created an unprecedented situation where the medical community and the public may not be familiar with the various new tests for COVID-19 that are offered to patients and hospitals. The purpose of this review is to provide information, up-to-date as of the date of submission of the manuscript to the journal, on the various tests that have been developed, their scientific basis and their interpretation. We give a real-world example demonstrating the time lag in the return of test results and review testing prioritization guidance since the supply of tests remains below the perceived need.


title: Adenovirus and miRNAs

text: Abstract Adenovirus infection has a tremendous impact on the cellular silencing machinery. Adenoviruses express high amounts of non-coding virus associated (VA) RNAs able to saturate key factors of the RNA interference (RNAi) processing pathway, such as Exportin 5 and Dicer. Furthermore, a proportion of VA RNAs is cleaved by Dicer into viral microRNAs (mivaRNAs) that can saturate Argonaute, an essential protein for miRNA function. Thus, processing and function of cellular miRNAs is blocked in adenoviral-infected cells. However, viral miRNAs actively target the expression of cellular genes involved in relevant functions such as cell proliferation, DNA repair or RNA regulation. Interestingly, the cellular silencing machinery is active at early times post-infection and can be used to control the adenovirus cell cycle. This is relevant for therapeutic purposes against adenoviral infections or when recombinant adenoviruses are used as vectors for gene therapy. Manipulation of the viral genome allows the use of adenoviral vectors to express therapeutic miRNAs or to be silenced by the RNAi machinery leading to safer vectors with a specific tropism. This article is part of a "Special Issue entitled:MicroRNAs in viral gene regulation".


title: Self-reactive CD4(+) T cells activated during viral-induced demyelination do not prevent clinical recovery

text: BACKGROUND: Microbial infections have been implicated in initiating and enhancing severity of autoimmune diseases including the demyelinating disease multiple sclerosis (MS). Nevertheless, the incidence of both acute and persisting viral infections without evidence of autoimmune sequelae suggests that this process is well controlled. The conditions promoting or stemming self-reactive (SR) T cells following viral-induced tissue damage thus need to be better defined. Using a non-fatal viral mouse model of encephalomyelitis associated with demyelination and disability, yet ultimate clinical improvement, this study set out to monitor uptake and presentation of endogenous myelin antigens, as well as induction and fate of SR T cells. METHODS: Activation and central nervous system (CNS) recruitment of myelin-specific CD4 T cells was analyzed by flow cytometry during encephalomyelitis induced by a glia tropic murine coronavirus. Potential antigen-presenting cells (APC) ingesting myelin were characterized by flow cytometry and their ability to activate SR T cells tested by co-culture with carboxyfluorescein succinimidyl ester (CFSE)-labeled myelin-specific CD4 T cells. Endogenous SR T cell kinetics was analyzed within both cervical lymph nodes and CNS by Enzyme-Linked ImmunoSpot (ELISPOT) following viral infection. RESULTS: The data demonstrate the presence of APC capable of activating SR T cells in both draining lymph nodes and the CNS temporally correlating with overt demyelination. While both the CNS-infiltrating myeloid population and microglia ingested myelin, only CNS-infiltrating APC were capable of presenting endogenous myelin antigen to SR T cells ex vivo. Finally, SR T cell activation from the endogenous T cell repertoire was most notable when infectious virus was controlled and paralleled myelin damage. Although SR T cell accumulation peaked in the persistently infected CNS during maximal demyelination, they were not preferentially retained. Their gradual decline, despite ongoing demyelination, suggested minimal re-stimulation and pathogenic function in vivo consistent with the lack of autoimmune symptoms. CONCLUSIONS: The results demonstrate the potential for CNS tissue destruction to induce and recruit SR T cells to the injury site and support a host suppressive mechanism limiting development of autoimmunity.


title: Viral Pneumonia in Patients with Hematopoietic Cell Transplantation and Hematologic Malignancies

text: Viral pneumonia is a common complication for patients with hematologic malignancies and after hematopoietic cell transplantation causing significant morbidity, and often mortality. Infections are predominantly caused by herpes viruses, either by reactivation of latent infection, or less commonly primary infection, or community respiratory viruses. High-resolution CT scan is useful for diagnosis but is nonspecific; generally, bronchoalveolar lavage is required. Prevention strategies are not pathogen-specific but include vaccination, chemoprophylaxis, preemptive treatment, and effective infection-prevention strategies during community outbreaks. Directed antiviral treatment is available for some pathogens. Toxicities and viral resistance are perennial challenges.


Can a disease be conquered by extensive publications, reading guidelines and interminable meetings?


Management of advanced melanoma in the COVID‐19 era


title: Lessons learned from COVID‐19 outbreak in a skilled nursing facility, Washington State

text: Long‐term care facilities have been identified as a local epicenter of disease among populations vulnerable to coronavirus disease 2019 (COVID‐19). A skilled nursing facility in Washington State was the first major site of COVID‐19 infections in the United States. Many lessons were learned during the events surrounding this outbreak, including how to develop, and the importance of, a coordinated response between emergency medical services and local area hospitals. As these events came early in the U.S. pandemic, unfortunately, disease spread and mortality was high. However, these events also resulted in rapid mobilization of the regional response to the COVID‐19 pandemic. Understanding the events surrounding this outbreak demonstrate some of the challenges involved in responding to acute infectious illnesses within these unique environments and associated vulnerable populations.


title: Microarray Analysis of Gene Expression of Cancer to Guide the Use of Chemotherapeutics

text: Summary The beauty of microarray analysis of gene expression (MAGE) is that it can be used to discover some genes that were previously thought to be unrelated to a physiologic or pathologic event. During the period from 1999 to 2007, applications of MAGE in cancer investigation have shifted from molecular profiling, identifying previously undiscovered cancer types, predicting outcomes of cancer patients, revealing metastasis signatures of solid tumors, to guiding the use of therapeutics. The roles of cancer genomic signatures have evolved through three phases. In the first phase, genomic signatures were described in stored cancer specimens and dubbed as molecular portraits of cancer. When gene expression profiles were carefully correlated with sufficient clinical information of cancer patients, new subgroups of cancers with distinct outcomes were revealed. In studies of the second phase, validation of cancer signatures was emphasized and commonly performed with independent groups of cancer specimens or independent data set. In the third phase, cancer genomic signatures have been further expanded beyond depicting the molecular portrait of cancer to predicting patient outcomes and guiding the use of cancer therapeutics. Cancer genomic signatures have become an essential part of a new generation of cancer clinical trials. It is advocated that, in future clinical trials of cancer therapy, the cancer specimens of each participant should be tested for currently available predictor genomic signatures, so that the most effective treatment with the least adverse effects for each patient can be identified. Then, participants can be triaged to an appropriate study group.


title: Epidemiological and Clinical Characteristics of COVID-19 in Adolescents and Young Adults

text: Summary Background Adolescents and young adults might play a key role in the worldwide spread of Coronavirus Disease 2019 (COVID-19) because they are more likely to be involved in overseas study, business, work, and travel. However, the epidemiological and clinical characteristics remain unknown. Methods We collected demographic, epidemiological, and clinical data from 46 confirmed COVID-19 patients aged 10 to 35 years from the Chongqing Three Gorges Central Hospital. Several key epidemiological parameters, asymptomatic cases, transmission to family members, and clinical characteristics at admission and during treatment were summarized. Results Of 46 confirmed patients, 14 patients (30.4%) were aged between 10 and 24 years, and 24 (52.2%) patients were male. The estimated mean incubation period was 6.6 days (95% confidence interval [CI] 4.4–9.6). The median serial interval was 1.9 days (95% CI 0.4–6.2). Three of the asymptomatic cases showed transmission to their family members. Only one patient was identified as a severe case at admission. The common symptoms at admission were dry cough (34, 81.0%) and fever (29, 69.1%). Nearly 60% of the patients showed ground-glass opacity on chest computed tomography. Three patients developed acute kidney injury during treatment. Most of the patients (78.3%) recovered and were discharged by the end of the follow-up. Conclusions This single-center study with a relatively small sample size showed that adolescent and young adult patients with COVID-19 had a long incubation period and a short serial interval. The transmission occurred from asymptomatic cases to family members. Fewer patients developed complications during treatment.


Tests Covid-19 : applications collectives et individuelles


title: The dynamics of risk perceptions and precautionary behavior in response to 2009 (H1N1) pandemic influenza

text: BACKGROUND: The trajectory of an infectious disease outbreak is affected by the behavior of individuals, and the behavior is often related to individuals' risk perception. We assessed temporal changes and geographical differences in risk perceptions and precautionary behaviors in response to H1N1 influenza. METHODS: 1,290 US adults completed an online survey on risk perceptions, interests in pharmaceutical interventions (preventive intervention and curative intervention), and engagement in precautionary activities (information seeking activities and taking quarantine measures) in response to H1N1 influenza between April 28 and May 27 2009. Associations of risk perceptions and precautionary behaviors with respondents' sex, age, and household size were analyzed. Linear and quadratic time trends were assessed by regression analyses. Geographic differences in risk perception and precautionary behaviors were evaluated. Predictors of willingness to take pharmaceutical intervention were analyzed. RESULTS: Respondents from larger households reported stronger interest in taking medications and engaged in more precautionary activities, as would be normatively predicted. Perceived risk increased over time, whereas interest in pharmaceutical preventive interventions and the engagement in some precautionary activities decreased over time. Respondents who live in states with higher H1N1 incidence per population perceived a higher likelihood of influenza infection, but did not express greater interests in pharmaceutical interventions, nor did they engage in a higher degree of precautionary activities. Perceived likelihood of influenza infection, willingness to take medications and engagement in information seeking activities were higher for women than men. CONCLUSIONS: Perceived risk of infection and precautionary behavior can be dynamic in time, and differ by demographic characteristics and geographical locations. These patterns will likely influence the effectiveness of disease control measures.


title: Selective Packaging in Murine Coronavirus Promotes Virulence by Limiting Type I Interferon Responses

text: Selective packaging is a mechanism used by multiple virus families to specifically incorporate genomic RNA (gRNA) into virions and exclude other types of RNA. Lineage A betacoronaviruses incorporate a 95-bp stem-loop structure, the packaging signal (PS), into the nsp15 locus of ORF1b that is both necessary and sufficient for the packaging of RNAs. However, unlike other viral PSs, where mutations generally resulted in viral replication defects, mutation of the coronavirus (CoV) PS results in large increases in subgenomic RNA packaging with minimal effects on gRNA packaging in vitro and on viral titers. Here, we show that selective packaging is also required for viral evasion of the innate immune response and optimal pathogenicity. We engineered two distinct PS mutants in two different strains of murine hepatitis virus (MHV) that packaged increased levels of subgenomic RNAs, negative-sense genomic RNA, and even cellular RNAs. All PS mutant viruses replicated normally in vitro but caused dramatically reduced lethality and weight loss in vivo. PS mutant virus infection of bone marrow-derived macrophages resulted in increased interferon (IFN) production, indicating that the innate immune system limited the replication and/or pathogenesis of PS mutant viruses in vivo. PS mutant viruses remained attenuated in MAVS(−/−) and Toll-like receptor 7-knockout (TLR7(−/−)) mice, two well-known RNA sensors for CoVs, but virulence was restored in interferon alpha/beta receptor-knockout (IFNAR(−/−)) mice or in MAVS(−/−) mice treated with IFNAR-blocking antibodies. Together, these data indicate that coronaviruses promote virulence by utilizing selective packaging to avoid innate immune detection.


title: Recombinase Polymerase Amplification for Diagnostic Applications

text: BACKGROUND: First introduced in 2006, recombinase polymerase amplification (RPA) has stirred great interest, as evidenced by 75 publications as of October 2015, with 56 of them just in the last 2 years. The widespread adoption of this isothermal molecular tool in many diagnostic fields represents an affordable (approximately 4.3 USD per test), simple (few and easy hands-on steps), fast (results within 5–20 min), and sensitive (single target copy number detected) method for the identification of pathogens and the detection of single nucleotide polymorphisms in human cancers and genetically modified organisms. CONTENT: This review summarizes the current knowledge on RPA. The molecular diagnostics of various RNA/DNA pathogens is discussed while highlighting recent applications in clinical settings with focus on point-of-care (POC) bioassays and on automated fluidic platforms. The strengths and limitations of this isothermal method are also addressed. SUMMARY: RPA is becoming a molecular tool of choice for the rapid, specific, and cost-effective identification of pathogens. Owing to minimal sample-preparation requirements, low operation temperature (25–42 °C), and commercial availability of freeze-dried reagents, this method has been applied outside laboratory settings, in remote areas, and interestingly, onboard automated sample-to-answer microfluidic devices. RPA is undoubtedly a promising isothermal molecular technique for clinical microbiology laboratories and emergence response in clinical settings.


title: Pathology, Molecular Biology, and Pathogenesis of Avian Influenza A (H5N1) Infection in Humans

text: H5N1 avian influenza is a highly fatal infectious disease that could cause a potentially devastating pandemic if the H5N1 virus mutates into a form that spreads efficiently among humans. Recent findings have led to a basic understanding of cell and organ histopathology caused by the H5N1 virus. Here we review the pathology of H5N1 avian influenza reported in postmortem and clinical studies and discuss the key pathogenetic mechanisms. Specifically, the virus infects isolated pulmonary epithelial cells and causes diffuse alveolar damage and hemorrhage in the lungs of infected patients. In addition, the virus may infect other organs, including the trachea, the intestines, and the brain, and it may penetrate the placental barrier and infect the fetus. Dysregulation of cytokines and chemokines is likely to be one of the key mechanisms in the pathogenesis of H5N1 influenza. We also review the various molecular determinants of increased pathogenicity that have been identified in recent years and the role of avian and human influenza virus receptors in relation to the transmissibility of the H5N1 virus. A comprehensive appreciation of H5N1 influenza pathogenetic mechanisms should aid in the design of effective strategies for prevention, diagnosis, and treatment of this emerging disease.


title: Fast-track ventilation strategy to cater for pandemic patient isolation surges

text: Summary Background The shortage of isolation facilities in hospitals was highlighted during the severe acute respiratory syndrome (SARS) pandemic in 2003. Yet, as the nature and scale of future pandemics cannot be adequately estimated, it is difficult to justify construction of sufficient isolation facilities. A fast-track and cost-effective ventilation strategy for the retrofitting of existing general wards could help hospitals deal with patient surges. Aim This article reviews the effectiveness of a fast-track, makeshift isolation approach employed during the SARS outbreak which involved installing simple window-mounted exhaust fans to create negative-pressure airflow in hospital general wards. Methods Computational fluid dynamics (CFD) was used to assess by simulation whether the approach adopted meets US Centers for Disease and Control and Prevention requirements for properly constructed isolation wards. Findings CFD simulation revealed that this makeshift approach could match the ventilation standards of isolation rooms. The approach was certainly effective as no secondary infections were reported in hospitals that used it during SARS. Conclusions When there is a shortfall in isolation facilities to accommodate a surge in patients, the proposed ventilation set-up could be quickly and widely implemented by existing general wards.


title: Characterization of an efficient coronavirus ribosomal frameshifting signal: Requirement for an RNA pseudoknot

text: Abstract The genomic RNA of the coronavirus IBV contains an efficient ribosomal frameshifting signal at the junction of two overlapping open reading frames. We have defined by deletion analysis an 86 nucleotide sequence encompassing the overlap region which is sufficient to allow frameshifting in a heterologous context. The upstream boundary of the signal consists of the sequence UUUAAAC, which is the likely site of ribosomal slippage. We show by creation of complementary nucleotide changes that the RNA downstream of this “slippery” sequence folds into a tertiary structure termed a pseudoknot, the formation of which is essential for efficient frameshifting.


title: Comparison of the 5′ Leader Sequences of North American Isolates of Reference and Field Strains of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)

text: The 5′ leader is documented to be an important regulatory element in many (+) ssRNA virus genome. To understand the significance of the 5′ leader RNA of PRRSV, we determined the complete leader sequences of fifteen different North American strains of PRRSV and predicted their secondary structures. Viruses analysed included three reference strains and nine field strains originating from different geographic locations. To further examine the leader region, one of the field strains was adapted to grow in tissue culture, and three clones were isolated. We also predicted the secondary structures of two European strains based on their published sequences. The predicted RNA secondary structures of the leader sequences suggested the existence of three conserved domains formed by the 5′ region of the leader among the North American strains, two of which were conserved in the European strains. A variable structural domain was predicted from the 3′ region of the leader sequences of the North American strains, where all tissue culture-adapted isolates were characterized by a stem-loop while field isolates were characterized by an internal bulge within the stem-loop.


title: Evaluation of the NucliSens® Basic Kit assay for detection of Norwalk virus RNA in stool specimens

text: Abstract Norwalk-like viruses (NLVs) are a genetically diverse group of human caliciviruses that are the most common cause of epidemic gastroenteritis and are detected typically in stool by reverse transcription (RT)-PCR or electron microscopy (EM). The application of a rapid nucleic acid sequence-based amplification (NASBA) assay for the detection of NLV RNA in stool is described using the NucliSens® Basic Kit. Primers and probes for the NLV Basic Kit assay were based on the RNA polymerase region of the prototype NLV, Norwalk virus (NV) genome and could consistently detect 104 RT-PCR detectable units of NV RNA in a stool filtrate. When compared directly with RT-PCR on a dilution series of NV stool filtrate, the NucliSens® Basic Kit assay was equally sensitive. Cross-reactivity studies with a representative panel of other enteric pathogens were negative. When applied to 15 stool specimens from NV-challenged volunteers, the NASBA Basic Kit application for NV detection yielded 100% sensitivity, 50% specificity, and 67% concordance, using RT-PCR as the ‘gold standard’. Despite the specificity of the NASBA primer/probe sequences for NV, other representatives from both NLV genogroups I and II could be detected by the Basic Kit assay in outbreak stool specimens, although the results were inconsistent. Our results suggest that the NucliSens® Basic Kit assay provides a rapid and sensitive alternative to RT-PCR for detecting NV RNA in stool specimens. However, improvements in test specificity and primer design will be needed before the assay can be used routinely in the clinical setting.


Portal vein thrombosis in a patient with COVID-19


title: Challenges in the Management of patients with systemic light chain (AL) amyloidosis during the COVID‐19 pandemic

text: The SARS‐CoV‐2‐associated disease (COVID‐19) is primarily manifested as a respiratory tract infection but may affect and cause complications from multiple organ systems (cardiovascular, gastrointestinal, kidneys, hematopoietic and immune systems) while no proven specific therapy exists. The challenges associated with COVID‐19 are even greater for patients with light chain (AL) amyloidosis, a rare multisystemic disease affecting the heart, kidneys, liver, gastrointestinal and nervous system. Patients with AL amyloidosis may need to receive chemotherapy, which probably increases infection risk. Management of COVID‐19 may be particularly challenging in patients with AL amyloidosis who often present with cardiac dysfunction, nephrotic syndrome, neuropathy, low blood pressure and gastrointestinal symptoms. In addition, AL patients may be more susceptible to toxicities of drugs used to manage COVID‐19. Access to health care may be difficult or limited, diagnosis of AL amyloidosis may be delayed with detrimental consequences, treatment administration may need modification. Both patients and treating physicians need to adapt in a new reality.


Covid-19 associated autoimmune thrombotic thrombocytopenic purpura: Report of a case


title: Feasibility of parental collected nasal swabs for virus detection in young children with cystic fibrosis

text: Abstract Background The detrimental role of viruses has been well described in CF, although the pattern of virus infections has not been investigated in a longitudinal study. The primary aim was to determine the feasibility of fortnightly parent collected swabs in young children with CF. Methods Children under three years with CF were recruited. Nasal swabs were collected by parents every fortnight and during periods of symptoms over 12months. Nasal swabs were posted and virus detected using real-time PCR. Results Only 27% of the patients completed the study to 10months, although 98% of the swabs returned were adequate for analysis. Mould was observed growing on 23% of the returned swabs. There was no evidence to demonstrate relationships with symptoms and viruses, prolonged symptoms, prolonged shedding or patterns of virus infections. Conclusions This study highlights the need to further investigate the role of viruses in children with CF using a robust method of frequent collection in children for a longitudinal study, with appropriate storage and shipping techniques to avoid mould growth or other potential contaminants.


title: Reorganization of the endoscopic activity of Cancer Institutes during phase II of the Covid-19 emergency

text: After the lockdown during the emergency phase of the Covid-19 pandemic, we have to deal with phase 2, a period of uncertain duration, with a controlled and progressive return to normalization, in which we need to reconcile our work and our movements with the presence of the virus on our territory. Digestive endoscopic activity is a high-risk transmission procedure for Covid-19. The measures put in place to protect healthcare personnel and patients are stressful and “time-consuming" and lead to a reduction in the number of endoscopic procedures that can be performed. In this scenario, the Oncological Institutes are forced to make a rigorous selection of patients to undergo endoscopic examinations and treatments, according to lists of exceptional priorities, in order to guarantee cancer patients and subjects at high risk of developing digestive tumors, a preferential diagnostic and therapeutic process, protected from contagion risks. For this purpose, cuts and postponing times of endoscopic performances are here proposed, which go beyond the guidelines of scientific societies and have little evidences in the literature. These changes should be applied limited to this exceptional period and in proportion to the capacity of each operating unit in order to meet the demands of the patients.


title: Canine parvovirus—A review of epidemiological and diagnostic aspects, with emphasis on type 2c

text: Abstract Canine parvovirus type 2 (CPV-2) emerged in late 1970s causing severe epizootics in kennels and dog shelters worldwide. Soon after its emergence, CPV-2 underwent genetic evolution giving rise consecutively to two antigenic variants, CPV-2a and CPV-2b that replaced progressively the original type. In 2000, a new antigenic variant, CPV-2c, was detected in Italy and rapidly spread to several countries. In comparison to the original type CPV-2, the antigenic variants display increased pathogenicity in dogs and extended host range, being able to infect and cause disease in cats. Epidemiological survey indicate that the newest type CPV-2c is becoming prevalent in different geographic regions and is often associated to severe disease in adult dogs and also in dogs that have completed the vaccination protocols. However, the primary cause of failure of CPV vaccination is interference by maternally derived immunity. Diagnosis of CPV infection by traditional methods has been shown to be poorly sensitive, especially in the late stages of infections. New diagnostic approaches based on molecular methods have been developed for sensitive detection of CPV in clinical samples and rapid characterisation of the viral type. Continuous surveillance will help assess whether there is a real need to update currently available vaccines and diagnostic tests.


title: Regional outcomes of severe acute respiratory syndrome coronavirus 2 infection in hospitalised patients with haematological malignancy

text: OBJECTIVES: We sought to characterise the outcomes of patients with haematological malignancy and SARS‐CoV‐2 infection in hospital in our regional network of 7 hospitals. METHODS: Consecutive hospitalised patients with haematological malignancy and SARS‐CoV‐2 infection were identified from 01/03/2020 to 06/05/2020. Outcomes were categorised as death, resolved, or ongoing. The primary outcome was preliminary case fatality rate (pCFR), defined as the number of cases resulting in death as a proportion of all diagnosed cases. Analysis was primarily descriptive. RESULTS: 66 Patients were included, overall pCFR was 51.5%. Patients ≥ 70 years accounted for the majority of hospitalised cases (42, 63%) and fatalities (25, 74%). Mortality was similar between females (52%) and males (51%). Immunosuppressive or cytotoxic treatment within 3 months of the diagnosis of SARS‐CoV‐2 infection was associated with a significantly higher preliminary Case Fatality Rate (pCFR) of 70%, compared with 28% in those not on active treatment (P = 0.0013, 2 proportions z‐test). CONCLUSIONS: Mortality rates in patients with haematological malignancy and SARS‐CoV‐2 infection in hospital are high supporting measures to minimise the risk of infection in this population.