title: Plant‐made vaccines for humans and animals

text: The concept of using plants to produce high‐value pharmaceuticals such as vaccines is 20 years old this year and is only now on the brink of realisation as an established technology. The original reliance on transgenic plants has largely given way to transient expression; proofs of concept for human and animal vaccines and of efficacy for animal vaccines have been established; several plant‐produced vaccines have been through Phase I clinical trials in humans and more are scheduled; regulatory requirements are more clear than ever, and more facilities exist for manufacture of clinic‐grade materials. The original concept of cheap edible vaccines has given way to a realisation that formulated products are required, which may well be injectable. The technology has proven its worth as a means of cheap, easily scalable production of materials: it now needs to find its niche in competition with established technologies. The realised achievements in the field as well as promising new developments will be reviewed, such as rapid‐response vaccines for emerging viruses with pandemic potential and bioterror agents.


title: Chapter 10 Advax Adjuvant A Potent and Safe Immunopotentiator Composed of Delta Inulin

text: Abstract Highly potent adjuvants commonly have reactogenicity and safety issues. Unlike other immunologically active polysaccharides, inulin has no immune activity in its native soluble form, but when crystallized into the stable delta inulin isoform, it acquires potent immune actions. Most notably, delta inulin enhances both humoral and cellular immune responses against vaccine antigens, translating into enhanced protection against a broad range of viral, bacterial, parasitic, and toxin-mediated diseases. Delta inulin is distinct compared with typical vaccine adjuvants as it does not require adsorption to antigen and does not appear to work through induction of inflammatory danger signals. Delta inulin manufactured under current Good Manufacturing Practices (referred to as Advax) has been proved to be safe and effective in human trials of influenza vaccines, hepatitis B vaccines, and insect sting allergy vaccines. Advax thereby represents a novel nonreactogenic immune modulator with potential utility across a broad range of human prophylactic and therapeutic vaccines.


title: Membrane topology of gp41 and amyloid precursor protein: Interfering transmembrane interactions as potential targets for HIV and Alzheimer treatment

text: Abstract The amyloid precursor protein (APP), that plays a critical role in the development of senile plaques in Alzheimer disease (AD), and the gp41 envelope protein of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome (AIDS), are single-spanning type-1 transmembrane (TM) glycoproteins with the ability to form homo-oligomers. In this review we describe similarities, both in structural terms and sequence determinants of their TM and juxtamembrane regions. The TM domains are essential not only for anchoring the proteins in membranes but also have functional roles. Both TM segments contain GxxxG motifs that drive TM associations within the lipid bilayer. They also each possess similar sequence motifs, positioned at the membrane interface preceding their TM domains. These domains are known as cholesterol recognition/interaction amino acid consensus (CRAC) motif in gp41 and CRAC-like motif in APP. Moreover, in the cytoplasmic domain of both proteins other α-helical membranotropic regions with functional implications have been identified. Recent drug developments targeting both diseases are reviewed and the potential use of TM interaction modulators as therapeutic targets is discussed.


Melatonina, coronavirus, enfermedad cardiovascular y emergencia geriátrica: ¡usemos todo lo que tenemos! Respuesta


title: Comparing the Binding Interactions in the Receptor Binding Domains of SARS-CoV-2 and SARS-CoV

text: [Image: see text] SARS-CoV-2, since emerging in Wuhan, China, has been a major concern because of its high infection rate and has left more than six million infected people around the world. Many studies endeavored to reveal the structure of the SARS-CoV-2 compared to the SARS-CoV, in order to find solutions to suppress this high infection rate. Some of these studies showed that the mutations in the SARS-CoV spike (S) protein might be responsible for its higher affinity to the ACE2 human cell receptor. In this work, we used molecular dynamics simulations and Monte Carlo sampling to compare the binding affinities of the S proteins of SARS-CoV and SARS-CoV-2 to the ACE2. Our results show that the protein surface of the ACE2 at the receptor binding domain (RBD) exhibits negative electrostatic potential, while a positive potential is observed for the S proteins of SARS-CoV/SARS-CoV-2. In addition, the binding energies at the interface are slightly higher for SARS-CoV-2 because of enhanced electrostatic interactions. The major contributions to the electrostatic binding energies result from the salt bridges forming between R426 and ACE-2-E329 in the case of SARS-CoV and K417 and ACE2-D30 in the SARS-CoV-2. In addition, our results indicate that the enhancement in the binding energy is not due to a single mutant but rather because of the sophisticated structural changes induced by all these mutations together. This finding suggests that it is implausible for the SARS-CoV-2 to be a lab-engineered virus.


title: Snapshot of the evolution and mutation patterns of SARS-CoV-2

text: The COVID-19 pandemic is the most important public health threat in recent history. Here we study how its causal agent, SARS-CoV-2, has diversified genetically since its first emergence in December 2019. We have created a pipeline combining both phylogenetic and structural analysis to identify possible human-adaptation related mutations in a data set consisting of 4,894 SARS-CoV-2 complete genome sequences. Although the phylogenetic diversity of SARS-CoV-2 is low, the whole genome phylogenetic tree can be divided into five clusters/clades based on the tree topology and clustering of specific mutations, but its branches exhibit low genetic distance and bootstrap support values. We also identified 11 residues that are high-frequency substitutions, with four of them currently showing some signal for potential positive selection. These fast-evolving sites are in the non-structural proteins nsp2, nsp5 (3CL-protease), nsp6, nsp12 (polymerase) and nsp13 (helicase), in accessory proteins (ORF3a, ORF8) and in the structural proteins N and S. Temporal and spatial analysis of these potentially adaptive mutations revealed that the incidence of some of these sites was declining after having reached an (often local) peak, whereas the frequency of other sites is continually increasing and now exhibit a worldwide distribution. Structural analysis revealed that the mutations are located on the surface of the proteins that modulate biochemical properties. We speculate that this improves binding to cellular proteins and hence represents fine-tuning of adaptation to human cells. Our study has implications for the design of biochemical and clinical experiments to assess whether important properties of SARS-CoV-2 have changed during the epidemic.


title: Characterization of enterotoxigenic E. coli (ETEC), Shiga-toxin producing E. coli (STEC) and necrotoxigenic E. coli (NTEC) isolated from diarrhoeic Mediterranean water buffalo calves (Bubalus bubalis)

text: Abstract Two hundred and twenty Escherichia coli isolates from 314 Mediterranean water buffalo calves less than 4weeks old affected by severe diarrhoea with a lethal outcome were characterized for the presence of the virulence factors LT, ST, Stx1, Stx2, haemolysins, intimin, CNF1, CNF2, CDT-I, CDT-II, CDT-III, CDT-IV, and F17-related fimbriae (F17a, F17b, F17c, F17d). The prevalence of ETEC, STEC and NTEC were 1.8%, 6.8% and 20.9%, respectively. The ETEC isolates were all LT-positive and ST-negative. The STEC isolates were all Stx and intimin-positive, with Stx1 (80%) more frequent than Stx2 (27%). The NTEC isolates were all CNF and Hly-positive, with CNF2 (83%) more frequent than CNF1 (22%). Susceptibility assays to 11 antimicrobials displayed high rates of resistance (>30%) to antimicrobials tested. These data show that the most prevalent strains in diarrhoeic water buffalo calves were NTEC, mostly CNF2 and HlyA-positive, with strong associations CNF2/CDT-III and CNF2/F17c.


title: Public preferences for interventions to prevent emerging infectious disease threats: a discrete choice experiment

text: OBJECTIVE: When faced with an emergent epidemic with high mortality and morbidity potential, policy makers must decide what public health interventions to deploy at different stages of the outbreak. However, almost nothing is known about how the public view these interventions or how they trade off risks (of disease) with inconvenience (of interventions). In this paper, we aim to understand public perceptions on pandemic interventions, as well as to identify if there are any distinct respondent preference classes. DESIGN: A discrete choice experiment. SETTING: This study was fielded in Singapore between November 2012 and February 2013. PARTICIPANTS: A random sample of 500 Singapore residents aged 21 and over, including 271 women and 229 men, was analysed. OUTCOME MEASURES: Demographic information was collected from each participant. Participants were also shown a series of pairs of alternatives, each combining interventions and morbidity, mortality and cost outcomes and declared a preference for one combination. A random utility model was developed to determine the individual’s preference for interventions and a hierarchical cluster analysis was performed to identify distinct respondent preference classes. RESULTS: On average, participants preferred more intense interventions, and preferred scenarios with fewer deaths and lower tax. The number of infections did not significantly influence respondents’ responses. We identified two broad classes of respondents: those who were mortality averse and those who were expenditure averse. Education was found to be a predictor of group membership. CONCLUSION: Overall, there was considerable support for government interventions to prevent or mitigate outbreaks of emerging infectious diseases, including those that greatly restricted individual liberties, as long as the restrictions showed a reasonable chance of reducing the adverse health effects of the outbreak.


title: Rat poison and food security in the People’s Republic of China: focus on tetramethylene disulfotetramine (tetramine)

text: The last several years have seen a large number of mass poisonings in mainland China, particularly those caused by illicit rodenticides. One rat poison, tetramine (tetramethylene disulfotetramine) is responsible for a great percentage of death and injury in the People’s Republic of China (PRC). Tetramine is an acutely toxic substance with human oral toxicity estimated as low as 0.1 mg/kg, and is widely available in open markets in mainland China—this despite being prohibited for manufacture or sale in the PRC. Being a GABA antagonist, and having an extremely potent effect on the brain stem, many victims can quickly fall into convulsions and die within hours following ingestion. With no known effective antidote at this time, clinical data from the PRC show that acute cases of tetramine poisoning are extremely difficult to treat. The widespread use of tetramine—including its reported sale at a Malaysian outdoor market in September 2002—could exacerbate its hazard to public health, particularly in those areas having large overseas Chinese populations.


title: Immunity to PRRSV: Double-edged sword

text: Abstract The immune system is a double-edged sword for porcine reproductive and respiratory syndrome virus (PRRSV) infection. On one edge PRRSV has a predilection for immune cells and the disease manifestations can be linked directly to changes in the immune system. PRRSV appears to replicate extensively, if not exclusively, in cells of the immune lineage, notably macrophages; the direct replication of which may lead to immunosuppression, precipitate secondary infection and/or mediate disease. On the other edge, the virus stimulates immunity post-infection that protects an animal from re-infection. A vast array of structural and functionally distinct antibody specific to PRRSV are generated following infection or vaccination. Discrete populations of functional antibodies appear at different times and possibly reflect reactivity to different PRRSV polypeptides. Cell-mediated immune responses specific to PRRSV can be detected in various exposed pigs as well. Thus, the immune system appears to be intimately involved in both the disease process and protection from disease. It is unclear at this state of understanding what immune compartment provides protective immunity. Is it humoral (i.e. antibodies), selective functionally distinct populations of antibodies specific for selected PRRSV polypeptides or is cellular immunity essential for protection, or both. This review will attempt to summarize the current state of knowledge of the complex interaction of the immune system and PRRSV.


title: Alabama university to study emotions, behavior and COVID‐19

text: A psychology researcher at the University of Alabama was awarded a federal grant to study how people's emotions influence their response to the spread of the novel coronavirus. “Right now, there is a tremendous amount of uncertainty in day‐to‐day life around the globe,” Philip Gable, Ph.D., associate professor of psychology, said in a March 30 university news release. “We are interested in the role uncertainty plays in how people feel and behave, and we are especially interested to see how non‐conscious emotions toward avoiding the virus influence health behaviors.” Gable is leading the project awarded nearly $65,000 under the National Science Foundation's (NSF's) Rapid Response Research funding program, which is set up to review and fund projects with an urgent need to gather data during or right after emergency events and natural disasters. The research at UA is part of the NSF's effort to quickly start nonmedical, nonclinical care research that can be used immediately to explore how to model and understand the spread of COVID‐19, to inform and educate about the science of virus transmission and prevention and to encourage the development of processes and actions to address this global challenge. The spreading COVID‐19 in the United States creates a social psychological problem of a scale never encountered in modern times, the release stated. The findings will be published and the anonymous data from the app will be publicly available.


title: Telehealth in Uro-oncology Beyond the Pandemic: Toll or Lifesaver?

text: BACKGROUND: Telehealth services are rapidly embraced in uro-oncology due to the current coronavirus disease 2019 (COVID-19) pandemic. OBJECTIVE: To determine patients’ perspective on adoption of telehealth as a response to the pandemic and its sustainability in the future. DESIGN, SETTING, AND PARTICIPANTS: Following a COVID-19 outbreak, 101 patients with advanced genitourinary cancers are currently managed “virtually” for therapy administration at our tertiary care unit. They were surveyed about the current situation, and current and long-term employment of telehealth. INTERVENTION: Rapid implementation of virtual patient management. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Patients’ perception of anxiety of COVID-19 and cancer, perspective on telehealth measures as a reaction to the current COVID-19 pandemic, and long-term acceptance were used as outcomes. Wilcoxon matched-pair signed rank test, chi-square test, and Mann-Whitney U test were performed. RESULTS AND LIMITATIONS: Of 101 patients, 92 answered the questionnaire, with 71 (77.2%) responding virtually by e-mail or phone call. Anxiety of cancer (6/10, interquartile range [IQR] 3–8) superseded that of COVID-19 (four/10, IQR 2–5.25, p < 0.001), and patients oppose temporary treatment interruption. Of the patients, 66.0% perceive their susceptibility to COVID-19 as equal to or lower than the general population and 52.2% believe that COVID-19 will not affect their therapy. In future, patients (62.6%) prefer to maintain in-person appointments as opposed to complete remote care, but accept remote care during the pandemic (eight/10, IQR 5–9). Beyond the crisis, maintaining telehealth has low preference rates (four/10, IQR 2–7), with high acceptance for external laboratory controls (60.9%) and online visit management (48.9%), but lower acceptance for remote treatment planning including staging discussions (44.6%) and for referral to secondary care oncologists (17.4%). CONCLUSIONS: Despite the pandemic, cancer remains the key concern and patients are not willing to compromise on their treatment. Rapid implementation of telehealth is tolerated well during the need of social distancing, with a clear “red line” concerning changes in existing patient-physician relationships. Balancing future implementation of telehealth while considering patients’ demand for personal relationships will ensure human dignity in uro-oncology. PATIENT SUMMARY: We queried patients with genitourinary cancers treated in an almost virtual setting following a local coronavirus outbreak. Acceptance of telehealth during the current situation is high; however, long-term implementation of the adapted services is less favored. We deduce that patient-physician relationship is crucial for cancer patients and needs to be balanced against measures for social distancing to forge the future management.


title: Viral–bacterial co-infections in the respiratory tract

text: Preceding or concurrent viral respiratory tract infection can predispose to secondary bacterial co-infection throughout the airway. The mechanisms by which viruses promote these superinfections are diverse and replete. Whereas we understand much as to how viruses damage the airway and dysregulate both innate and acquired immune responses which, in turn, supports bacterial growth, adherence and invasion into normally sterile sites within the respiratory tract, new information regarding these co-infections is being gained from recent advances in microbiome research and our enhanced appreciation of the contribution of bacterial biofilms, among others. The advanced understanding obtained by continued research efforts in all aspects of viral–bacterial co-infections of the respiratory tract will allow us to devise novel approaches for disease prevention as well as to develop more effective therapeutics.


title: Implementing fit testing for N95 filtering facepiece respirators: Practical information from a large cohort of hospital workers

text: Qualitative fit testing for N95 respirators was conducted on 1271 health care workers. All male participants were fitted with a respirator. Six females, all under age 40 years, were not successfully fitted. The first-choice respirator provided a successful fit in 95.1% of the men and 85.4% of the women. Gender and age in women were significant factors associated with successful fitting. These differences should be considered when implementing a respiratory protection program.


title: Isolation and characterization of porcine epidemic diarrhea virus associated with the 2014 disease outbreak in Mexico: case report

text: BACKGROUND: Interest in porcine epidemic diarrhea has grown since the 2013 outbreak in the United States caused major losses, with mortality rates up to 100 % in suckling piglets. In Mexico, an outbreak of porcine epidemic diarrhea, characterized by 100 % mortality in piglets, began in March 2014 in the State of Mexico. METHODS: The aim of this study was to confirm and identify porcine epidemic diarrhea virus (PEDV) in samples from piglets with suggestive clinical signs using virological, histological, and molecular techniques. Necropsy was performed on 13 piglets from two litters with initial and advanced clinical signs. Suggestive lesions of acute infection with PEDV were detected in histological sections of the small and large bowels; specifically, multiple virus particles with visible crown-shaped projections were observed using electron microscopy and negative staining. Viral isolation was performed in Vero cells with trypsin. Infection was monitored by observation of cytopathic effect, and titration was determined by TCID(50)/ml. The presence of the PEDV in cultures and clinical samples was confirmed by RT-PCR amplification and sequencing of a 651-bp segment of the S glycoprotein gene, as well as a 681-bp matrix protein gene. RESULTS: The nucleotide sequence analysis of the Mexican isolates showed marked homology to viruses that circulated in 2013 in Colorado, USA. CONCLUSIONS: In this paper we confirm the isolation and characterization of PEDV from animals with early and advanced clinical signs.


title: Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70

text: BACKGROUND: Hepatitis B virus (HBV) infection is a major health concern with more than two billion individuals currently infected worldwide. Because of the limited effectiveness of existing vaccines and drugs, development of novel antiviral strategies is urgently needed. Heat stress cognate 70 (Hsc70) is an ATP-binding protein of the heat stress protein 70 family. Hsc70 has been found to be required for HBV DNA replication. Here we report, for the first time, that combined siRNAs targeting viral gene and siHsc70 are highly effective in suppressing ongoing HBV expression and replication. METHODS: We constructed two plasmids (S1 and S2) expressing short hairpin RNAs (shRNAs) targeting surface open reading frame of HBV(HBVS) and one plasmid expressing shRNA targeting Hsc70 (siHsc70), and we used the EGFP-specific siRNA plasmid (siEGFP) as we had previously described. First, we evaluated the gene-silencing efficacy of both shRNAs using an enhanced green fluorescent protein (EGFP) reporter system and flow cytometry in HEK293 and T98G cells. Then, the antiviral potencies of HBV-specific siRNA (siHBV) in combination with siHsc70 in HepG2.2.15 cells were investigated. Moreover, type I IFN and TNF-α induction were measured by quantitative real-time PCR and ELISA. RESULTS: Cotransfection of either S1 or S2 with an EGFP plasmid produced an 80%–90% reduction in EGFP signal relative to the control. This combinational RNAi effectively and specifically inhibited HBV protein, mRNA and HBV DNA, resulting in up to a 3.36 log(10) reduction in HBV load in the HepG2.2.15 cell culture supernatants. The combined siRNAs were more potent than siHBV or siHsc70 used separately, and this approach can enhance potency in suppressing ongoing viral gene expression and replication in HepG2.2.15 cells while forestalling escape by mutant HBV. The antiviral synergy of siHBV used in combination with siHsc70 produced no cytotoxicity and induced no production of IFN-α, IFN-β and TNF-α in transfected cells. CONCLUSIONS: Our combinational RNAi was sequence-specific, effective against wild-type and mutant drug-resistant HBV strains, without triggering interferon response or producing any side effects. These findings indicate that combinational RNAi has tremendous promise for developing innovative therapy against viral infection.


title: In silico/In vivo analysis of high-risk papillomavirus L1 and L2 conserved sequences for development of cross-subtype prophylactic vaccine

text: Human papillomavirus (HPV) is the most common sexually transmitted infection in the world and the main cause of cervical cancer. Nowadays, the virus-like particles (VLPs) based on L1 proteins have been considered as the best candidate for vaccine development against HPV infections. Two commercial HPV (Gardasil and Cervarix) are available. These HPV VLP vaccines induce genotype-limited protection. The major impediments such as economic barriers especially gaps in financing obstructed the optimal delivery of vaccines in developing countries. Thus, many efforts are underway to develop the next generation of vaccines against other types of high-risk HPV. In this study, we developed DNA constructs (based on L1 and L2 genes) that were potentially immunogenic and highly conserved among the high-risk HPV types. The framework of analysis include (1) B-cell epitope mapping, (2) T-cell epitope mapping (i.e., CD4(+) and CD8(+) T cells), (3) allergenicity assessment, (4) tap transport and proteasomal cleavage, (5) population coverage, (6) global and template-based docking, and (7) data collection, analysis, and design of the L1 and L2 DNA constructs. Our data indicated the 8-epitope candidates for helper T-cell and CTL in L1 and L2 sequences. For the L1 and L2 constructs, combination of these peptides in a single universal vaccine could involve all world population by the rate of 95.55% and 96.33%, respectively. In vitro studies showed high expression rates of multiepitope L1 (~57.86%) and L2 (~68.42%) DNA constructs in HEK-293T cells. Moreover, in vivo studies indicated that the combination of L1 and L2 DNA constructs without any adjuvant or delivery system induced effective immune responses, and protected mice against C3 tumor cells (the percentage of tumor-free mice: ~66.67%). Thus, the designed L1 and L2 DNA constructs would represent promising applications for HPV vaccine development.


title: Full-length genome analysis of two genetically distinct variants of porcine epidemic diarrhea virus in Thailand

text: Abstract Porcine epidemic diarrhea virus (PEDV) has continued to cause sporadic outbreaks in Thailand since 2007 and a pandemic variant containing an insertion and deletion in the spike gene was responsible for outbreaks. In 2014, there were further outbreaks of the disease occurring within four months of each other. In this study, the full-length genome sequences of two genetically distinct PEDV isolates from the outbreaks were characterized. The two PEDV isolates, CBR1/2014 and EAS1/2014, were 28,039 and 28,033 nucleotides in length and showed 96.2% and 93.6% similarities at nucleotide and amino acid levels respectively. In total, we have observed 1048 nucleotide substitutions throughout the genome. Compared to EAS1/2014, CBR1/2014 has 2 insertions of 4 (56GENQ59) and 1 (140N) amino acid positions 56–59 and 140, and 2 deletions of 2 (160DG161) and 1 (1199Y) amino acid positions 160–161 and 1199. The phylogenetic analysis based on full-length genome of CBR1/2014 isolate has grouped the virus with the pandemic variants. In contrast, EAS1/2014 isolate was grouped with CV777, LZC and SM98, a classical variant. Our findings demonstrated the emergence of EAS1/2014, a classical variant which is novel to Thailand and genetically distinct from the currently circulating endemic variants. This study warrants further investigations into molecular epidemiology and genetic evolution of the PEDV in Thailand.


10 Adenovirus Vaccines


Treating hypoxemic COVID-19 “ARDS” patients with almitrine: The earlier the better?


title: Zinc: a new risk factor for pneumonia in the elderly?

text: Zinc may be a new risk factor for pneumonia in the elderly. In this special article, we reviewed the magnitude of the problem of pneumonia (its prevalence, morbidity and mortality) in the elderly, its etiology, and the dysregulation of the immune system associated with increasing age. In addition, we presented evidence from the literature, including work we did recently, that low zinc status (commonly reported in the elderly) impairs immune function, decreases resistance to pathogens, and is associated not only with increased incidence and duration of pneumonia, increased use and duration of antimicrobial treatment, but also with increased overall mortality in the elderly. Inadequate stores of zinc might therefore be a risk factor for pneumonia in the elderly. Randomized, double blind, controlled studies are needed to determine the efficacy of zinc supplementation as a potential low cost intervention to reduce morbidity and mortality due to pneumonia in this vulnerable population.


title: Timeliness of infectious disease reporting, the Netherlands, 2003 to 2017: law change reduced reporting delay, disease identification delay is next

text: BACKGROUND: Timely notification of infectious diseases is essential for effective disease control and needs regular evaluation. AIM: Our objective was to evaluate the effects that statutory adjustments in the Netherlands in 2008 and raising awareness during outbreaks had on notification timeliness. METHODS: In a retrospective analyses of routine surveillance data obtained between July 2003 and November 2017, delays between disease onset and laboratory confirmation (disease identification delay), between laboratory confirmation and notification to Municipal Health Services (notification delay) and between notification and reporting to the National Institute for Public Health and the Environment (reporting delay) were analysed for 28 notifiable diseases. Delays before (period 1) and after the law change (periods 2 and 3) were compared with legal timeframes. We studied the effect of outbreak awareness in 10 outbreaks and the effect of specific guidance messages on disease identification delay for two diseases. RESULTS: We included 144,066 notifications. Average notification delay decreased from 1.4 to 0.4 days across the three periods (six diseases; p < 0.05), reporting delay decreased mainly in period 2 (from 0.5 to 0.1 days, six diseases; p < 0.05). In 2016–2017, legal timeframes were met overall. Awareness resulted in decreased disease identification delay for three diseases: measles and rubella (outbreaks) and psittacosis (specific guidance messages). CONCLUSIONS: Legal adjustments decreased notification and reporting delays, increased awareness reduced identification delays. As disease identification delay dominates the notification chain, insight in patient, doctor and laboratory delay is necessary to further improve timeliness and monitor the impact of control measures during outbreaks.


title: Prospective and retrospective evaluation of the Cepheid Xpert® Flu/RSV XC assay for rapid detection of influenza A, influenza B, and respiratory syncytial virus

text: Abstract A total of 281 clinical specimens (nasal swabs and nasopharyngeal aspirates) were tested with the Xpert® Flu/RSV XC. The results were compared to those obtained with the real-time retro transcriptase-polymerase chain reaction assays routinely used in our laboratory. The Xpert® Flu/RSV XC showed sensitivity/specificity of 97.8%/100% and 97.9%/100% for flu and respiratory syncytial virus, respectively.


title: An investigation of district spatial variations of childhood diarrhoea and fever morbidity in Malawi

text: Although diarrhoea and malaria are among the leading causes of child mortality and morbidity in Sub-Saharan Africa, few detailed studies have examined the patterns and determinants of these ailments in the most affected communities. In this paper, we investigate the spatial distribution of observed diarrhoea and fever prevalence in Malawi using individual data for 10,185 children from the 2000 Malawi Demographic and Health survey. We highlight inequalities in child health by mapping the residual district spatial effects using a geo-additive probit model that simultaneously controls for spatial dependence in the data and potential nonlinear effects of covariates. The residual spatial effects were modelled via a Bayesian approach. For both ailments, we were able to identify a distinct district pattern of childhood morbidity. In particular, the results suggest that children living in the capital city are less affected by fever, although this is not true for diarrhoea, where some urban agglomerations are associated with a higher childhood morbidity risk. The spatial patterns emphasize the role of remoteness as well as climatic, environmental, and geographic factors on morbidity. The fixed effects show that for diarrhoea, the risk of child morbidity appears to be lower among infants who are exclusively breastfed than among those who are mixed-fed. However, exclusive breastfeeding was not found to have a protective effect on fever. An important socio-economic factor for both diarrhoea and fever morbidity was parental education, especially maternal educational attainment. Diarrhoea and fever were both observed to show an interesting association with child's age. We were able to discern the continuous worsening of the child morbidity up to 8–12 months of age. This deterioration set in right after birth and continues, more or less linearly until 8–12 months, before beginning to decline thereafter. Independent of other factors, a separate spatial process produces district inequalities in child's health.


title: An Observational Cohort Study of Hydroxychloroquine and Azithromycin for COVID-19: (Can’t Get No) Satisfaction

text: It can be said that SARS-CoV-2 caught the world by surprise. In large part due to globalization, the virus quickly evolved from a serious regional concern to a worldwide pandemic, the likes of which are unprecedented in the last century. In a matter of weeks, COVID-19 became a leading cause of death, with a potential staggering death toll in 2020. Due to a heavy burden of illness, and in the absence of proven therapies, several experimental treatments have been and continue to be prescribed outside of clinical trial settings. Of the potential therapeutic options that showed early promise, few have generated as much controversy, or been subject to such politicization, as hydroxychloroquine. In this issue of the International Journal of Infectious Diseases, Arshad et. al have added more fuel to the fire.


title: Comparison of two broadly multiplexed PCR systems for viral detection in clinical respiratory tract specimens from immunocompromised children

text: Abstract Background The detection of viral respiratory tract infections has evolved greatly with the development of PCR based commercial systems capable of simultaneously detecting a wide variety of pathogens. Objectives Evaluate the relative performance of two commercial broad range systems for the detection of viral agents in clinical respiratory tract specimens from immunocompromised children. Study design A total of 176 patient samples were included in the analysis, representing only the first sample collected for each patient, and excluding failed reactions. Samples were de-identified and assayed in parallel using two different, broadly multiplexed PCR systems: ResPlex™ II Panel v2.0 (ResPlex), Qiagen, Hilden, Germany and FilmArray® Respiratory Panel (FilmArray), Idaho Technology Inc., Salt Lake City, UT. Method comparison was based upon pair-wise concordance of results according to patient age, viral target and number of targets detected. Results The two systems showed an overall concordance, by patient, of 83.8% (p =0.0001). FilmArray detected at least one target in 68.8% of samples, while ResPlex detected at least one target in 56.8%. ResPlex failed to detect 20.7% of FilmArray positives, and FilmArray failed to detect 4% of ResPlex positives. The relative performance of each system (including which system detected a higher number of positive samples) varied when stratified by target viral pathogen. Conclusions Broadly multiplexed PCR is an effective means of detecting large numbers of clinically relevant respiratory viral pathogens.


title: Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina

text: Cryptosporidiosis of calves is caused by the enteroprotozoan Cryptosporidium spp. The disease results in intense diarrhea of calves associated with substantial economic losses in dairy farming worldwide. The aim of this study was to determine calf, herd, and within-herd Cryptosporidium prevalence and identify Cryptosporidium species and subtypes in calves with diarrhea in intensive dairy herds in central Argentina. A total of 1073 fecal samples were collected from 54 randomly selected dairy herds. Cryptosporidium-oocysts were isolated and concentrated from fecal samples using formol-ether and detected by light microscopy with the modified Ziehl-Neelsen technique. Overall prevalence of oocyst-excreting calves was found to be 25.5% (274/1073) (95% C.I. 22.9; 28.1%). Of the herds studied, 89% (48/54) included at least one infected calf, whereas within-herd prevalence ranged from the absence of infection to 57% (20/35). A highly significant association was found between the presence of diarrhea and C. parvum infection (χ(2) = 55.89, p < 0.001). For species determination, genomic DNA isolated from oocyst-positive fecal samples was subjected to PCR-RFLP of the 18S rRNA gene resulting exclusively in Cryptosporidium parvum identification. C. parvum isolates of calves displaying diarrhea and high rate of excretion of oocysts were subtyped by PCR amplification and direct sequencing of the 60 kDa glycoprotein (GP60) gene. Altogether five GP60 subtypes, designated IIaA18G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1, and IIaA24G1R1 were identified. Interestingly, IIaA18G1R1 and IIaA20G1R1 were predominant in calves with diarrhea and high infection intensity. Notably, IIaA24G1R1 represents a novel, previously unrecognized C. parvum subtype. The subtype IIaA18G1R1, frequently found in this study, is strongly implicated in zoonotic transmission. These results suggest that calves might be an important source for human cryptosporidiosis in Argentina. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00436-019-06366-y) contains supplementary material, which is available to authorized users.


title: Murine cortical brain cells are autoantigenic from a distinct development stage onwards

text: Abstract The expression of autoantigens on murine cortical brain cells and their first appearance during development was studied. Autoreactivity was analyzed by weight increase and lymphocyte proliferation in the popliteal lymph node (PLN). Cortical brain cells or defined plasma membrane preparations were injected s.c. without adjuvant into syngeneic recipients. Weak, but significant T cell-dependent PLN enlargement was triggered with brain cells from adult mice. A stronger reaction could be elicited with one defined fraction of purified plasma membranes. The earliest appearance of the antigenic material in the plasma membrane fraction was observed on day 15 after birth. This time point correlates exactly with the completion of the blood-brain barrier in large parts of the central nervous system.


title: Respiratory viral coinfection and disease severity in children: A systematic review and meta-analysis

text: Abstract Background With advent of molecular diagnostic technologies, studies have reported detection of two or more respiratory viruses in about 30% of children with respiratory infections. However, prognostic role of coinfection remains unclear. Objective Evaluate relation between respiratory viral confection and illness severity in children. Study design MEDLINE (through PUBMED), EMBASE, EBSCO, LILACS databases were searched up to March 2015 by two independent reviewers. Studies assessing severity of viral coinfection in patients aged less than 18 years were included. Standardized forms were used for data extraction of population, study design, clinical syndromes, virus combinations compared and severity outcomes. Risk of bias and quality of evidence were assessed through EPHPP and GRADE. Subgroup analysis was performed according to age and viral combinations. Results Of 5218 records screened, 43 were included in analysis. Viral coinfection did not influence risks of all outcomes assessed: length of stay (mean difference in days in coinfection, −0.10 [95% confidence interval: −0.51 to 0.31]), length of supplemental oxygen (−0.42 [−1.05 to 0.20]), need of hospitalization (odds ratio of coinfection, 0.96 [95% confidence interval: 0.61–1.51]), supplemental oxygen (0.94 [0.66 to 1.34]), need of intensive care (0.99 [0.64 to 1.54]), mechanical ventilation (0.81 [0.33 to 2.01]) and death (2.22 [0.83 to 5.95]). Sub-analyses according to age and viral combinations have not shown influence of these factors in outcomes. Conclusions Respiratory viral coinfection did not increase severity in all outcomes assessed. Further studies are necessary to confirm this finding, especially regarding role of specific viral interactions.


title: Population-Level Effect of Cholera Vaccine on Displaced Populations, South Sudan, 2014

text: Following mass population displacements in South Sudan, preventive cholera vaccination campaigns were conducted in displaced persons camps before a 2014 cholera outbreak. We compare cholera transmission in vaccinated and unvaccinated areas and show vaccination likely halted transmission within vaccinated areas, illustrating the potential for oral cholera vaccine to stop cholera transmission in vulnerable populations.


title: LGBTQ+ Inclusive Palliative Care in the Context of COVID-19:Pragmatic Recommendations for Clinicians

text: As Coronavirus Disease 2019 (COVID-19) continues to impact the seriously ill and their families on a global scale, considerations given to marginalized groups amid the pandemic is essential to ensure the provision of high-quality, dignified care. Lesbian, gay, bisexual, transgender, gender-nonconforming, queer and/or questioning (LGBTQ+) identified persons are particularly vulnerable to health inequities across settings, including palliative care and at end-of-life. There is a crucial gap in the literature pertaining to palliative care during COVID-19. We aim to fill this gap by providing essential health inequity and social support background pertaining to LGBTQ+ persons and practical recommendations for immediate implementation that support inclusive and respectful care for these populations. Employing these recommendations is a pragmatic pathway to promote trust, transparency, patient and family engagement, and value concordant care amid the health system strain caused by COVID-19.


title: Tetrabutylammonium fluoride-assisted rapid N(9)-alkylation on purine ring: Application to combinatorial reactions in microtiter plates for the discovery of potent sulfotransferase inhibitors in situ

text: Tremendous efforts have been invested in the synthesis of purine libraries due to their importance in targeting various enzymes involved in different diseases and cellular processes. The synthesis of N(9)-alkylated purine scaffolds relied mostly on Mitsunobu conditions with a variety of alcohols or strong basic conditions with different organic halides. A more reliable and efficient way for the synthesis of N(9)-alkylated purine scaffolds is reported. This method uses tetrabutylammonium fluoride (TBAF) to assist such chemistry. In many cases, the reactions were completed within 10 min and gave the desired product in high yield and selectivity. Moreover, these mild reaction conditions permitted its use in combinatorial reactions in microtiter plates followed by in situ screening for the discovery of potent sulfotransferase inhibitors.


title: Acute cholecystitis during COVID-19 pandemic: a multisocietary position statement

text: Following the spread of the infection from the new SARS-CoV2 coronavirus in March 2020, several surgical societies have released their recommendations to manage the implications of the COVID-19 pandemic for the daily clinical practice. The recommendations on emergency surgery have fueled a debate among surgeons on an international level. We maintain that laparoscopic cholecystectomy remains the treatment of choice for acute cholecystitis, even in the COVID-19 era. Moreover, since laparoscopic cholecystectomy is not more likely to spread the COVID-19 infection than open cholecystectomy, it must be organized in such a way as to be carried out safely even in the present situation, to guarantee the patient with the best outcomes that minimally invasive surgery has shown to have.


title: Honey Bee Queen Production: Canadian Costing Case Study and Profitability Analysis

text: The decline in managed honey bee (Hymenoptera: Apidae) colony health worldwide has had a significant impact on the beekeeping industry. To mitigate colony losses, beekeepers in Canada and around the world introduce queens into replacement colonies; however, Canada’s short queen rearing season has historically limited the production of early season queens. As a result, Canadian beekeepers rely on the importation of foreign bees, particularly queens from warmer climates. Importing a large proportion of (often mal-adapted) queens each year creates a dependency on foreign bee sources, putting beekeeping, and pollination sectors at risk in the event of border closures, transportation issues, and other restrictions as is currently happening due to the 2020 Covid-19 pandemic. Although traditional Canadian queen production is unable to fully meet early season demand, increasing domestic queen production to meet mid- and later season demand would reduce Canada’s dependency. As well, on-going studies exploring the potential for overwintering queens in Canada may offer a strategy to have early season domestic queens available. Increasing the local supply of queens could provide Canadian beekeepers, farmers, and consumers with a greater level of agricultural stability and food security. Our study is the first rigorous analysis of the economic feasibility of queen production. We present the costs of queen production for three Canadian operations over two years. Our results show that it can be profitable for a beekeeping operation in Canada to produce queen cells and mated queens and could be one viable strategy to increase the sustainability of the beekeeping industry.


title: Gga-miR-30d regulates infectious bronchitis virus infection by targeting USP47 in HD11 cells

text: Abstract Avian infectious bronchitis virus (IBV) is a coronavirus which infects chickens and causes severe economic losses to the poultry industry worldwide. MicroRNAs (miRNAs) are important intracellular regulators and play a pivotal role in viral infections. In previous studies, we have revealed that IBV infection caused a significant down-regulation of gga-miR-30d expression in chicken kidneys. In present study, we investigated the role of gga-miR-30d in the process of IBV infection of HD11 cell line in vitro. By transfecting the mimics and inhibitor of gga-miR-30d, it was found that overexpressed gga-miR-30d inhibited IBV replication. Contrarily, low-expressed gga-miR-30d promoted IBV replication. In addition, dual-luciferase reporter assays revealed that ubiquitin-specific protease 47 (USP47), a deubiquitinase-encoding gene, was a target for gga-miR-30d. This is the first study demonstrating that miRNAs regulate IBV replication by regulating the deubiquitinating enzyme (DUBs).


title: The COVID-19 Infection in Italy: A Statistical Study of an Abnormally Severe Disease

text: We statistically investigate the Coronavirus Disease 19 (COVID-19) pandemic, which became particularly invasive in Italy in March 2020. We show that the high apparent lethality or case fatality ratio (CFR) observed in Italy, as compared with other countries, is likely biased by a strong underestimation of the number of infection cases. To give a more realistic estimate of the lethality of COVID-19, we use the actual (March 2020) estimates of the infection fatality ratio (IFR) of the pandemic based on the minimum observed CFR and analyze data obtained from the Diamond Princess cruise ship, a good representation of a “laboratory” case-study from an isolated system in which all the people have been tested. From such analyses, we derive more realistic estimates of the real extent of the infection as well as more accurate indicators of how fast the infection propagates. We then isolate the dominant factors causing the abnormal severity of the disease in Italy. Finally, we use the death count—the only data estimated to be reliable enough—to predict the total number of people infected and the interval of time when the infection in Italy could end.


title: Attenuated early inflammatory response in solid organ recipients with COVID‐19

text: Immunosuppression leaves transplanted patients at particular risk for severe acute respiratory syndrome 2 (SARS‐CoV‐2) infection. The specific features of coronavirus disease 2019 (COVID‐19) in immunosuppressed patients are largely unknown and therapeutic experience is lacking. Seven transplanted patients (two liver, three kidneys, one double lung, one heart) admitted to the Ludwig‐Maximilians‐University Munich because of COVID‐19 and tested positive for SARS‐CoV‐2 were included. The clinical course and the clinical findings were extracted from the medical record. The two liver transplant patients and the heart transplant patient had an uncomplicated course and were discharged after 14, 18 and 12 days, respectively. Two kidney transplant recipients were intubated within 48 hours. One kidney and the lung transplant recipients were required to intubate after ten and 15 days, respectively. Immunosuppression was adapted in five patients, but continued in all patients. Compared to non‐transplanted patients at the ICU (n=19) the inflammatory response was attenuated in transplanted patients, which was proven by decreased IL‐6 blood values. This analysis might provide evidence that continuous immunosuppression is safe and probably beneficial since there was no hyperinflammation evident. Although transplanted patients might be more susceptible to an infection with SARS‐CoV‐2, their clinical course seems to be similar to immunocompetent patients.


title: Epidemiology and genetic variability of respiratory syncytial virus in Portugal, 2014–2018

text: Abstract Introduction Respiratory syncytial virus (RSV) is associated with substantial morbidity and mortality since it is a predominant viral agent causing respiratory tract infections in infants, young children and the elderly. Considering the availability of the RSV vaccines in the coming years, molecular understanding in RSV is necessary. Objective The objective of the present study was to describe RSV epidemiology and genotype variability in Portugal during the 2014/15-2017/18 period. Material and methods Epidemiological data and RSV-positive samples from patients with a respiratory infection were collected through the non-sentinel and sentinel influenza surveillance system (ISS). RSV detection, subtyping in A and B, and sequencing of the second hypervariable region (HVR2) of G gene were performed by molecular methods. Phylogenetic trees were generated using the Neighbor-Joining method and p-distance model on MEGA 7.0. Results RSV prevalence varied between the sentinel (2.5%, 97/3891) and the non-sentinel ISS (20.7%, 3138/16779), being higher (P < 0.0001) among children aged <5 years. Bronchiolitis (62.9%, 183/291) and influenza-like illness (24.6%, 14/57) were associated (P < 0.0001) with RSV laboratory confirmation among children aged <6 months and adults ≥65 years, respectively. The HVR2 was sequenced for 562 samples. RSV-A (46.4%, 261/562) and RSV-B (53.6%, 301/562) strains clustered mainly to ON1 (89.2%, 233/261) and BA9 (92%, 277/301) genotypes, respectively, although NA1 and BA10 were also present until 2015/2016. Conclusion The sequence and phylogenetic analysis reflected the relatively high diversity of Portuguese RSV strains. BA9 and ON1 genotypes, which have been circulating in Portugal since 2010/2011 and 2011/2012 respectively, predominated during the whole study period.


title: Early gene expression events in ferrets in response to SARS coronavirus infection versus direct interferon-alpha2b stimulation

text: Abstract Type I interferons (IFNs) are essential to the clearance of viral diseases, however, a clear distinction between genes upregulated by direct virus–cell interactions and genes upregulated by secondary IFN production has not been made. Here, we investigated differential gene regulation in ferrets upon subcutaneous administration of IFN-α2b and during SARS-CoV infection. In vivo experiments revealed that IFN-α2b causes STAT1 phosphorylation and upregulation of abundant IFN response genes (IRGs), chemokine receptors, and other genes that participate in phagocytosis and leukocyte transendothelial migration. During infection with SARS-CoV not only a variety of IRGs were upregulated, but also a significantly broader range of genes involved in cell migration and inflammation. This work allowed dissection of several molecular signatures present during SARS-CoV which are part of a robust IFN antiviral response. These signatures can be useful markers to evaluate the status of IFN responses during a viral infection and specific features of different viruses.


title: Cross-species transmission potential between wild pigs, livestock, poultry, wildlife, and humans: implications for disease risk management in North America

text: Cross-species disease transmission between wildlife, domestic animals and humans is an increasing threat to public and veterinary health. Wild pigs are increasingly a potential veterinary and public health threat. Here we investigate 84 pathogens and the host species most at risk for transmission with wild pigs using a network approach. We assess the risk to agricultural and human health by evaluating the status of these pathogens and the co-occurrence of wild pigs, agriculture and humans. We identified 34 (87%) OIE listed swine pathogens that cause clinical disease in livestock, poultry, wildlife, and humans. On average 73% of bacterial, 39% of viral, and 63% of parasitic pathogens caused clinical disease in other species. Non-porcine livestock in the family Bovidae shared the most pathogens with swine (82%). Only 49% of currently listed OIE domestic swine diseases had published wild pig surveillance studies. The co-occurrence of wild pigs and farms increased annually at a rate of 1.2% with as much as 57% of all farms and 77% of all agricultural animals co-occurring with wild pigs. The increasing co-occurrence of wild pigs with livestock and humans along with the large number of pathogens shared is a growing risk for cross-species transmission.


SARS-cov-2 related deaths in French long-term care facilities: the “confinement disease” is probably more deleterious than the COVID-19 itself


title: Standardization of Laboratory Methods for the PERCH Study

text: The Pneumonia Etiology Research for Child Health study was conducted across 7 diverse research sites and relied on standardized clinical and laboratory methods for the accurate and meaningful interpretation of pneumonia etiology data. Blood, respiratory specimens, and urine were collected from children aged 1–59 months hospitalized with severe or very severe pneumonia and community controls of the same age without severe pneumonia and were tested with an extensive array of laboratory diagnostic tests. A standardized testing algorithm and standard operating procedures were applied across all study sites. Site laboratories received uniform training, equipment, and reagents for core testing methods. Standardization was further assured by routine teleconferences, in-person meetings, site monitoring visits, and internal and external quality assurance testing. Targeted confirmatory testing and testing by specialized assays were done at a central reference laboratory.


title: Results of magnetic resonance imaging in 14 cats with meningoencephalitis

text: Medical records and magnetic resonance (MR) images of 14 cats with inflammatory diseases affecting the central nervous system (CNS) were reviewed retrospectively. Cases included eight cats with feline infectious peritonitis and two cats with toxoplasmosis. Abnormalities affecting the CNS were observed in MR images in 10 (71%) cats. Intracranial lesions appeared as slightly hypointense foci in T1-weighted images in two (14%) cats, as hyperintense foci in T2-weighted images in seven (50%) cats and as hyperintense foci after intravenous administration of a gadolinium-based contrast medium in 10 (71%) cats. In six cats with lesions in T1- and/or T2-weighted images, additional lesions were visible in T1-weighted images obtained after gadolinium-based contrast medium administration. In three cats, lesions were visible only after contrast medium administration. In our study, MR imaging (MRI) did not appear to detect all cases of CNS inflammation in the population of cats with inflammatory cerebrospinal fluid (CSF); however, MRI adds information about the sites and morphology of intracranial lesions that should help to distinguish between neoplasia and inflammatory conditions and, possibly, between different inflammatory conditions.


title: The role of telemedicine in the delivery of healthcare in the COVID‐19 Pandemic

text: On 11 March 2020, the World Health Organization declared the coronavirus disease 2019 (COVID‐19) a pandemic. Recently, Hermans, et.al., outlined the challenges the COVID‐19 pandemic pose for the bleeding disorders community (1). The general response to the pandemic has included increased measures focused on personal hygiene, social distancing, symptom monitoring, early diagnosis, patient isolation, shelter in place, and public health quarantine. Accordingly, such measures have led to concerns over how to maintain access to haemophilia treatment products and to the specialized integrated care medical follow up within the haemophilia treatment centers (HTCs). As part of the medical response to disasters such as a world‐wide pandemic, telemedicine has emerged as one proposed solution to address this type of global challenge (2).


title: Treatment with convalescent plasma for COVID‐19 patients in Wuhan, China

text: The discovery of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and the outbreak of coronavirus disease 2019 (COVID‐19) are causing public health emergency. A handful of literatures have summarized its clinical and radiologic features, whereas therapies for COVID‐19 are rather limited. In order to evaluate the efficacy of convalescent plasma therapy in COVID‐19 patients, we did this timely descriptive study. 6 laboratory confirmed COVID‐19 patients were enrolled and received the transfusion of ABO‐compatible convalescent plasma. The efficacy of this intervention was determined by the alleviation of symptoms, changes in radiologic abnormalities and laboratory tests. No obvious adverse effect observed during the treatment. Transfusion of convalescent plasma led to a resolution of ground glass opacities (GGOs) and consolidation in patient #1, #2, #3, #4 and #6. In patient #1 and #5 who presented with SARS‐CoV‐2 in throat swab, convalescent plasma therapy elicited an elimination of virus. Serologic analysis indicated an immediate increase in anti‐SARS‐CoV‐2 antibody titers in patient #2 and #3, but not in patient #1. This study indicates that convalescent plasma therapy is effective and specific for COVID‐19. This intervention has a special significance for eliminating SARS‐CoV‐2 and is believed to be a promising state‐of‐art therapy during COVID‐19 pandemic crisis. This article is protected by copyright. All rights reserved.


title: Probable transmission chains of Middle East respiratory syndrome coronavirus and the multiple generations of secondary infection in South Korea

text: Summary Background In May 2015, South Korea reported its first case of Middle East respiratory syndrome coronavirus (MERS-CoV) infection in a 68-year-old man with a history of travel in the Middle East. In the presence of secondary infections, an understanding of the transmission dynamics of the virus is crucial. The aim of this study was to characterize the transmission chains of MERS-CoV infection in the current South Korean outbreak. Methods Individual-level data from multiple sources were collected and used for epidemiological analyses. Results As of July 14, 2015, 185 confirmed cases of MERS have been reported in the Korean outbreak. Three generations of secondary infection, with over half belonging to the second generation, could be delineated. Hospital infection was found to be the most important cause of virus transmission, affecting largely non-healthcare workers (154/184). Healthcare switching has probably accounted for the emergence of multiple generations of secondary infection. Fomite transmission may explain a significant proportion of the infections occurring in the absence of direct contact with infected cases. Conclusions Publicly available data from multiple sources, including the media, are useful to describe the epidemic history of an outbreak. The effective control of MERS-CoV hinges on the upholding of infection control standards and an understanding of health-seeking behaviours in the community.


title: Establishment of fetal bovine intestinal epithelial cell cultures susceptible to bovine rotavirus infection

text: Abstract Mucosal epithelial cells are infected by a wide variety of pathogens and determining their response to infection is critical for understanding disease pathogenesis. A protocol was developed for culturing primary epithelial cells from fetal bovine intestine and the cultured cells were evaluated for susceptibility to an enteric viral infection. Immunohistochemical staining for cytokeratin confirmed that 60–75% of cultured cells were epithelial cells. Furthermore, following infection with bovine rotavirus (BRV) over 80% of cells in the ileal and jejunal cultures contained viral protein at 16h post-infection. The intestinal epithelial cell cultures also contained fibroblasts so a jejunal fibroblast culture was established and infected with BRV. Viral protein was detected in jejunal fibroblasts but viral-induced cytopathology was delayed in fibroblast cultures when compared to epithelial cell cultures. This study describes an effective protocol for culturing bovine epithelial cells from fetal intestine and confirmed that the epithelial cells were susceptible to BRV infection. Ileal and jejunal cultures displayed limited growth following continuous passage but early passage epithelial cells provide competent target cells for studying host cell responses to an enteric viral pathogen.


title: Using an online learning module to teach nursing students about food insecurity as a social determinant of health

text: Currently, food insecurity is a lack of access to enough food for a healthy life that impacts approximately 40 million Americans and 821 million people worldwide. These statistics are expected to rise as a result of the COVID-19 pandemic drastically. Studies show that healthcare and nursing curricula are currently deficient in providing education to help healthcare providers combat food security and provide patient education. The purpose of this article is to discuss a curricular framework that can be implemented in both nursing and healthcare curricula to offer appropriate training for food insecurity assessment, care, and patient education.


title: VIRsiRNApred: a web server for predicting inhibition efficacy of siRNAs targeting human viruses

text: BACKGROUND: Selection of effective viral siRNA is an indispensable step in the development of siRNA based antiviral therapeutics. Despite immense potential, a viral siRNA efficacy prediction algorithm is still not available. Moreover, performances of the existing general mammalian siRNA efficacy predictors are not satisfactory for viral siRNAs. Therefore, we have developed “VIRsiRNApred” a support vector machine (SVM) based method for predicting the efficacy of viral siRNA. METHODS: In the present study, we have employed a new dataset of 1725 viral siRNAs with experimentally verified quantitative efficacies tested under heterogeneous experimental conditions and targeting as many as 37 important human viruses including HIV, Influenza, HCV, HBV, SARS etc. These siRNAs were divided into training (T(1380)) and validation (V(345)) datasets. Important siRNA sequence features including mono to penta nucleotide frequencies, binary pattern, thermodynamic properties and secondary structure were employed for model development. RESULTS: During 10-fold cross validation on T(1380) using hybrid approach, we achieved a maximum Pearson Correlation Coefficient (PCC) of 0.55 between predicted and actual efficacy of viral siRNAs. On V(345) independent dataset, our best model achieved a maximum correlation of 0.50 while existing general siRNA prediction methods showed PCC from 0.05 to 0.18. However, using leave one out cross validation PCC was improved to 0.58 and 0.55 on training and validation datasets respectively. SVM performed better than other machine learning techniques used like ANN, KNN and REP Tree. CONCLUSION: VIRsiRNApred is the first algorithm for predicting inhibition efficacy of viral siRNAs which is developed using experimentally verified viral siRNAs. We hope this algorithm would be useful in predicting highly potent viral siRNA to aid siRNA based antiviral therapeutics development. The web server is freely available at http://crdd.osdd.net/servers/virsirnapred/.


Infección por coronavirus (COVID-19) en Anales de Pediatría


title: Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune Sources

text: Antibodies are critical reagents in many fundamental biochemical methods such as affinity chromatography. As our understanding of the proteome becomes more complex, demand is rising for rapidly generated antibodies of higher specificity than ever before. It is therefore surprising that few investigators have moved beyond the classical methods of antibody production in their search for new reagents. Despite their long-standing efficacy, recombinant antibody generation technologies such as phage display are still largely the tools of biotechnology companies or research groups with a direct interest in protein engineering. In this chapter, we discuss the inherent limitations of classical polyclonal and monoclonal antibody generation and highlight an attractive alternative: generating high specificity, high affinity recombinant antibodies from alternative immune sources such as chickens, via phage display.


title: Prokaryotic expression, refolding, and purification of fragment 450–650 of the spike protein of SARS-coronavirus

text: Abstract The spike (S) glycoprotein is one of the major structure proteins of SARS-associated coronavirus (CoV). Fragment 450–650 (S450-650) of the S protein contains receptor-binding domain and neutralizing epitopes. In this study, S450-650 was expressed with a histidine tag in Escherichia coli BL21. Bacterial inclusion bodies containing the recombinant S450-650 were solubilized with 8M urea and then applied onto a Ni–nitrilotriacetic acid column. On-column refolding and purification was performed. Reduced glutathione and oxidized glutathione were included in the refolding buffer. In the wash and elution buffers, glycerol and glucose were necessary additives to prevent protein aggregation during purification. This refolding and purification procedure allowed production of S450-650 at up to 500μg/ml in soluble form, which maintained appropriate antigenicity and immunogenicity. It was able to induce strong IgG responses in BALB/c mice. In Western blot assays, the recombinant S450-650 was recognized by monoclonal Ab against the His-tag and also sera from a convalescent SARS patient. S450-650-based ELISA system was able to detect anti-SARS-CoV IgG Abs in patient sera.


title: Characteristics of a coronavirus causing vomition and wasting in pigs

text: Some characteristics of a virus, isolated from the tonsils of 2 pigs with clinical signs of inappetence and vomition and designated VW572 were examined. It induced the formation of syncytia in primary pig kidney cell cultures, caused hemadsorption and hemagglutination using chicken, turkey, rat and mouse erythrocytes. In growth curve experiments, infectious virus was produced intracellularly starting at 6 hours after inoculation and was followed by rapid release of the virus from the infected cells. The virus contains ribonucleic acid, is ether sensitive and has a size between 100 and 220 nm. At 37° C, the infectivity titer decreased about 2 log(10)TCID(50) per 24 hours. The viral population was heterogeneous as indicated by the rate of inactivation by U.V. irradiation and acid pH. Some hemagglutinating activity was left after complete loss of infectivity by ether-, temperature- and U.V. treatment. The VW 572 isolate is antigenically related if not identical to isolates from Canada, U.S.A. and England which were classified as a porcine coronavirus. Oronasal and intracerebral inoculation of the VW572 isolate in colostrum deprived pigs resulted in clinical disease after an incubation period of 6 and 4 days, respectively. Signs were characterized by depression, vomition, loss of appetence with rapid weakness in young pigs and vomition with progressive wasting in older pigs. Virus was isolated from nasal and pharyngeal swabs, nasal mucosa, tonsils, lungs and hind brains but not from rectal swabs or other organs tested. Virus could not be isolated later than 8 days after inoculation. Hemagglutination inhibiting and neutralizing antibodies were detected in sera at 6 days and 9 days after inoculation, respectively.


title: Enteric Colonization by Staphylococcus delphini in Four Ferret Kits with Diarrhoea

text: Summary Four, 1-to 4-week-old ferret kits were submitted to the Diagnostic Center for Population and Animal Health at Michigan State University for post-mortem examination. Grossly, multiple bowel loops in all ferret kits were distended by mucoid faecal material. Microscopically, there was no evidence of inflammation or notable alteration to the normal mucosal morphology. Gram-positive coccoid bacteria colonized variable segments of the small intestine. These bacteria were identified as Staphylococcus delphini by phenotypic and molecular analyses. Enzyme-linked immunosorbent assay for detection of Staphylococcus enterotoxins was positive and polymerase chain reaction detected the gene for Staphylococcus enterotoxin E in the isolates. The hypersecretory diarrhoea in these ferret kits may have been associated with colonization of the small intestine by S. delphini, cultures of which were shown in vitro to be potentially capable of producing enterotoxin E. The condition described in these ferrets is similar to ‘sticky’ kit syndrome in mink.


Three Decades of Interferon-β in Multiple Sclerosis: Can We Repurpose This Information for the Management of SARS-CoV2 Infection?


title: Synthesis and biological activity evaluation of 5-pyrazoline substituted 4-thiazolidinones

text: A series of novel 5-pyrazoline substituted 4-thiazolidinones have been synthesized. Target compounds were evaluated for their anticancer activity in vitro within DTP NCI protocol. Among the tested compounds, the derivatives 4d and 4f were found to be the most active, which demonstrated certain sensitivity profile toward the leukemia subpanel cell lines with GI(50) value ranges of 2.12–4.58 μM (4d) and 1.64–3.20 μM (4f). The screening of antitrypanosomal and antiviral activities of 5-(3-naphthalen-2-yl-5-aryl-4,5-dihydropyrazol-1-yl)-thiazolidine-2,4-diones was carried out with the promising influence of the mentioned compounds on Trypanosoma brucei, but minimal effect on SARS coronavirus and influenza types A and B viruses.


title: “Remote monitoring of intranasal ketamine self-administration as maintenance therapy in treatment-resistant depression (TRD): a novel strategy for vulnerable and at-risk populations to COVID-19?”

text: Abstract This work describes a clinical case in which we used telemedicine to monitor at-home ketamine treatment in TRD. Ketamine has been shown to be a promising and safe treatment for TRD in elderly patients. However, the COVID-19 pandemic is clearly impacting the world and this disease is recognized as being particularly dangerous for the elderly and vulnerable patients. To limit the transmission of the disease, geriatric mental health specialists must be creative and ensure the safety of their patients by limiting human to human contact. This problem might be averted by massive deployment of outreach services and telemedicine strategies.


title: p53 Acts as a Safeguard of Translational Control by Regulating Fibrillarin and rRNA Methylation in Cancer

text: Ribosomes are specialized entities that participate in regulation of gene expression through their rRNAs carrying ribozyme activity. Ribosome biogenesis is overactivated in p53-inactivated cancer cells, although involvement of p53 on ribosome quality is unknown. Here, we show that p53 represses expression of the rRNA methyl-transferase fibrillarin (FBL) by binding directly to FBL. High levels of FBL are accompanied by modifications of the rRNA methylation pattern, impairment of translational fidelity, and an increase of internal ribosome entry site (IRES)-dependent translation initiation of key cancer genes. FBL overexpression contributes to tumorigenesis and is associated with poor survival in patients with breast cancer. Thus, p53 acts as a safeguard of protein synthesis by regulating FBL and the subsequent quality and intrinsic activity of ribosomes.


title: Respiratory syncytial virus: an important cause of acute respiratory illness among young adults undergoing military training

text: Background Military recruits receiving training are vulnerable to acute respiratory disease and a significant proportion of illness is caused by unidentified pathogens. While some countries use surveillance programmes to monitor such illness, few data exist for recruits of the British Armed Forces. Objectives Through active surveillance of approximately 1000 Royal Navy trainees during 2001, we sought to describe and determine the aetiology of acute respiratory illness. Methods Standard viral culture was used together with serology and a novel highly sensitive real‐time PCR and molecular beacon probe assay for respiratory syncytial virus (RSV) detection. Results Among 54 Royal Navy recruits with respiratory symptoms adenovirus was identified in 35%, influenza viruses in 19% and RSV in 14%. All recruits were absent from training for almost a week, most of whom were confined to the sickbay. Conclusions This study is the first to document adenovirus and RSV as important causes of acute respiratory illness among Royal Navy trainees. The study findings demonstrate the clinical significance and challenges of diagnosing RSV infection in young adults.


title: What influenza vaccination programmes are preferred by healthcare personnel? A discrete choice experiment

text: OBJECTIVES: This study examined the relative importance of factors relating to vaccine characteristics, social normative influence and convenience in access to vaccine for determining decision making for seasonal influenza vaccination (SIV) among healthcare personnel (HCP), aiming to optimize existing influenza vaccination programmes for HCP. METHODS: A discrete choice experiment (DCE) was conducted in HCP working in public hospitals in Hong Kong. The DCE was designed to examine the relative importance of vaccine characteristics (vaccine efficacy and safety), social normative influence reflected by the proportion of HCP colleagues intending to take SIV, and convenience in access to vaccine indicated by vaccination programme duration, vaccination location, vaccination arrangement procedure and service hours in determining influenza vaccination choice among HCP. Mixed logit regression modelling was conducted to examine the preference weight (β) of factors included in the DCE for determining vaccination choice. RESULTS: Vaccination probability increased with increase in vaccine efficacy (β = 0.02 for per 1% increase), vaccination location changing from “designated staff clinic” to “mobile station” (β = 0.37), vaccination arrangement procedure changing from “by appointment” to “by walk-in” (β = 0.99), but decreased with the increase in probability of mild reactions to vaccination (β = −0.05 for per 1% increase). CONCLUSION: Vaccine safety was judged to be more important than vaccine efficacy for determining vaccination choice. Arranging vaccination service by walk-in and implementing mobile vaccination station should be considered in future SIV programmes to compensate for the effect of perceived low vaccination efficacy and concerns about vaccine safety to promote SIV uptake among HCP.


title: Enrichment of coronavirus-induced interferon-producing blood leukocytes increases the interferon yield per cell: A study with pig leukocytes

text: Summary Porcine peripheral blood mononuclear cells, which secrete IFNα in response to a coronavirus, transmissible gastroenteritis virus, were detected by a filter immunoplaque assay (ELISPOT). IFNα-producing cells (IPC), which are present at a low frequency in the blood, could be enriched up to 100-fold by sequential depletion of plastic-adherent cells and cell fractionation on metrizamide density gradients. IPC were present in the non-adherent low-density cell subpopulation. Cell selection experiments using antibody (Ab)-coated immunomagnetic beads revealed that porcine IPC could be positively selected by anti-CD4 or -SLA-class-II Ab, but not by anti-CD2 or -CD8 Ab. The estimated IFN yield per IPC was found to increase when IPC were assayed at higher concentrations. These data suggest that IPC represent a unique and distinct cell population in the blood, which could secrete higher amounts of IFN following its accumulation at a site of viral infection.


title: Acute Transverse Myelitis Associated with SARS-CoV-2: A Case-Report

text: • The third case of acute transverse myelitis due to SARS-CoV-2 reported in the world; • Possible inflammatory complications affecting the myelin in spinal cord. • We must be vigilant of the critical neurological illnesses associated with COVID-19.


Commentary: Nobody loves a critic: Edmund A Parkes and John Snow’s cholera


title: Tratamiento rehabilitador en la miositis aguda benigna infantil

text: Resumen La miositis aguda benigna infantil es un proceso poco frecuente, transitorio y autolimitado que afecta a niños en edad escolar (predominantemente chicos) tras una infección respiratoria de vías altas. Se caracteriza por el inicio súbito de dolor intenso en las extremidades inferiores y dificultad o imposibilidad para caminar con elevación sérica de niveles de creatinquinasa. Presentamos el caso de una niña de 8 años afecta de miositis aguda benigna infantil. Describimos el cuadro clínico, las claves para el diagnóstico diferencial con otros procesos más graves que asocian incapacidad para caminar y evaluamos los beneficios del tratamiento rehabilitador en los casos atípicos o complicados para facilitar la resolución del proceso y evitar las complicaciones. Abstract Benign acute childhood myositis (BACM) is a rare, temporary and self-limited condition, affecting school-age children (generally boys) after upper respiratory infection. It is characterized by sudden onset of severe pain in the legs and difficulty or inability to walk, with increase in serum creatine kinase (CK) levels. We present the case of an 8-year-old girl with BACM and describe the clinical picture, keys for the differential diagnosis with other more serious disorders that associate inability to walk. We evaluate the benefits of rehabilitation treatment in the atypical or complicated cases in order to provide the solution to the condition and avoid complications.


title: MULTICENTRE DOUBLE-BLIND STUDY OF EFFECT OF INTRATHECALLY ADMINISTERED NATURAL HUMAN FIBROBLAST INTERFERON ON EXACERBATIONS OF MULTIPLE SCLEROSIS

text: Abstract In this randomised, double-blind, placebo-controlled, 2-year multicentre study intrathecally administered natural human fibroblast interferon (IFN-B) was effective in reducing exacerbations of multiple sclerosis (MS) in patients with exacerbating/remitting disease. The mean reduction in exacerbation rate of 34 patients who received IFN-B (recipients) was significantly greater during the study than that of 35 patients who received placebo (p <0.04). The prestudy exacerbation rates were comparable in recipients and controls, but the rate at the end of the study was significantly lower in recipients than in controls (p <0.001). IFN-B was given by nine or ten lumbar punctures over the first 6 months of the study, and patient observations continued for 2 years. IFN-B was well tolerated in 95% of the recipients, and the side-effects experienced were clearly acceptable for the benefits achieved. Low doses of indomethacin reduced the toxicity of IFN-B and played an important role in successful double-blinding.


title: Public Health Response Systems In-Action: Learning from Local Health Departments’ Experiences with Acute and Emergency Incidents

text: As part of their core mission, public health agencies attend to a wide range of disease and health threats, including those that require routine, acute, and emergency responses. While each incident is unique, the number and type of response activities are finite; therefore, through comparative analysis, we can learn about commonalities in the response patterns that could improve predictions and expectations regarding the resources and capabilities required to respond to future acute events. In this study, we interviewed representatives from more than 120 local health departments regarding their recent experiences with real-world acute public health incidents, such as infectious disease outbreaks, severe weather events, chemical spills, and bioterrorism threats. We collected highly structured data on key aspects of the incident and the public health response, particularly focusing on the public health activities initiated and community partners engaged in the response efforts. As a result, we are able to make comparisons across event types, create response profiles, and identify functional and structural response patterns that have import for future public health preparedness and response. Our study contributes to clarifying the complexity of public health response systems and our analysis reveals the ways in which these systems are adaptive to the character of the threat, resulting in differential activation of functions and partners based on the type of incident. Continued and rigorous examination of the experiences of health departments throughout the nation will refine our very understanding of what the public health response system is, will enable the identification of organizational and event inputs to performance, and will allow for the construction of rich, relevant, and practical models of response operations that can be employed to strengthen public health systems.


Lung Recruitability in COVID-19–associated Acute Respiratory Distress Syndrome: A Single-Center Observational Study


title: Interdisciplinary and Collaborative Approaches Needed to Determine Impact of COVID-19 on Older Adults and Aging: CAG/ACG and CJA/RCV Joint Statement

text: The COVID-19 pandemic and subsequent state of public emergency have significantly affected older adults in Canada and worldwide. It is imperative that the gerontological response be efficient and effective. In this statement, the board members of the Canadian Association on Gerontology/L’Association canadienne de gérontologie (CAG/ACG) and the Canadian Journal on Aging/La revue canadienne du vieillissement (CJA/RCV) acknowledge the contributions of CAG/ACG members and CJA/RCV readers. We also profile the complex ways that COVID-19 is affecting older adults, from individual to population levels, and advocate for the adoption of multidisciplinary collaborative teams to bring together different perspectives, areas of expertise, and methods of evaluation in the COVID-19 response.


title: Prospective evaluation of an internet-linked handheld computer critical care knowledge access system

text: INTRODUCTION: Critical care physicians may benefit from immediate access to medical reference material. We evaluated the feasibility and potential benefits of a handheld computer based knowledge access system linking a central academic intensive care unit (ICU) to multiple community-based ICUs. METHODS: Four community hospital ICUs with 17 physicians participated in this prospective interventional study. Following training in the use of an internet-linked, updateable handheld computer knowledge access system, the physicians used the handheld devices in their clinical environment for a 12-month intervention period. Feasibility of the system was evaluated by tracking use of the handheld computer and by conducting surveys and focus group discussions. Before and after the intervention period, participants underwent simulated patient care scenarios designed to evaluate the information sources they accessed, as well as the speed and quality of their decision making. Participants generated admission orders during each scenario, which were scored by blinded evaluators. RESULTS: Ten physicians (59%) used the system regularly, predominantly for nonmedical applications (median 32.8/month, interquartile range [IQR] 28.3–126.8), with medical software accessed less often (median 9/month, IQR 3.7–13.7). Eight out of 13 physicians (62%) who completed the final scenarios chose to use the handheld computer for information access. The median time to access information on the handheld handheld computer was 19 s (IQR 15–40 s). This group exhibited a significant improvement in admission order score as compared with those who used other resources (P = 0.018). Benefits and barriers to use of this technology were identified. CONCLUSION: An updateable handheld computer system is feasible as a means of point-of-care access to medical reference material and may improve clinical decision making. However, during the study, acceptance of the system was variable. Improved training and new technology may overcome some of the barriers we identified.


title: Different altered stage correlative expression of high abundance acute-phase proteins in sera of patients with epithelial ovarian carcinoma

text: BACKGROUND: The general enhanced expression of α(1)-antichymotrypsin (ACT), clusterin (CLU), α(1)-antitrypsin (AAT), haptoglobin β-chain (HAP), and leucine rich glycoprotein (LRG) in the sera of patients with epithelial ovarian carcinoma (EOCa) was recently reported. In the present study, we compared the expression of the serum acute-phase proteins (APPs) in the patients according to their stages of cancer. RESULTS: Different altered stage correlative expression of the high abundance serum APPs was demonstrated in sera of the patients studied. While the expression of ACT, HAP and AAT appeared to demonstrate positive correlation with the three initial stages of the cancer, inverse correlation was apparently detected in the expression of LRG and CLU. For patients who were diagnosed with stage IV of the cancer, expression of the serum APPs did not conform to the altered progression changes. CONCLUSION: Our results highlight the potential prognostic significance of selective high abundance serum APPs in patients with EOCa.


title: Stimulation and analysis of the immune response in calves from vaccinated pregnant cows

text: Abstract The effect of vaccinating pregnant cows with an inactivated vaccine against Mannheimia haemolytica, BRSV and PI3V infections on selected immune responses in their offspring was examined. Blood samples were collected weekly for 12 weeks from six newborn calves from each of vaccinated (experimental) and unvaccinated (control) dams. Specific antibodies to M. haemolytica, BRSV and PI3V and mean values of IgA, IgG concentrations were significantly higher in the experimental calves compared with the controls. However, specific antibody titres to adenovirus type 3, BHV1 and BVDV in the experimental calves had constant levels while the control group levels changed. The IgM, Hp and SAA concentrations generally increased until week 8 in the experimental group, but the control group titres became higher after week 9. This study demonstrates that specific immunisation of cows pre-partum significantly stimulated parameters associated with immunity and it also controlled the acute phase response intensity in their offspring. Therefore the vaccination of dams may provide additional antibody protection against infection to their offspring.


Testing recommendation for COVID-19 (SARS-CoV-2) in patients planned for surgery - continuing the service and ‘suppressing’ the pandemic


title: CHAPTER 17 GUINEA PIGS

text: Publisher Summary This chapter deals with the health and medical care issues of guinea pigs. Guinea pigs have wide bodies with short limbs. They have a short, flat nose, laterally placed eyes, and hairless external pinnae. The dentition of the guinea pig is described as aradicular hypsodont. Guinea pigs are best housed in well-ventilated, wire-sided cages with solid bottoms. If housed indoors, guinea pig enclosures do not require a cover, as these animals do not typically jump or climb. Heavy food containers are recommended to make dumping of the receptacle more difficult. All food containers should be easy to disinfect and should be cleaned regularly, because guinea pigs have a habit of soiling their food bowls. These animals, native to the Andes Mountains, are very susceptible to hyperthermia and should never be housed in temperatures greater than 80°F. High humidity can also exacerbate a guinea pig's sensitivity to elevated temperatures by increasing the heat index. Guinea pigs often do not exhibit clinical signs early in a disease process. Therefore, a thorough physical examination can be extremely useful in determining the overall health status of the animal.


title: Coronavirus Immunoreactivity in Individuals With a Recent Onset of Psychotic Symptoms

text: Prenatal influenza exposure increases the risk for schizophrenia and brings to question how other respiratory viruses may contribute to neuropsychiatric disease etiopathology. Human coronaviruses cause respiratory infections that range in seriousness from common colds to severe acute respiratory syndrome. Like influenza, coronaviruses can be neurotropic. To test for associations between coronaviruses and serious mental disorders, we utilized a recently developed assay and measured immunoglobulin G (IgG) response against 4 human coronavirus strains (229E, HKU1, NL63, and OC43) in 106 patients with a recent onset of psychotic symptoms and 196 nonpsychiatric controls. We expressed results quantitatively as antibody levels and qualitatively as seroprevalence relative to a defined seropositivity cutoff value. Patient IgG levels were higher than controls for HKU1, NL63, and OC43, with HKU1 and NL63 both showing highly significant patient-to-control differences (HKU1, P ≤ .002; NL63, P ≤ .00001). All 4 coronaviruses were more seroprevalent in patients vs controls, with greatest intergroup differences observed for HKU1 (93% vs 77%, P ≤ .0001). HKU1 and NL63 associations with the patient group were further supported by multivariate analyses that controlled for age, gender, race, socioeconomic status, and smoking status (HKU1, odds ratio [OR] = 1.32, 95% confidence interval [CI] = 1.03–1.67, P ≤ .027; NL63, OR = 2.42, 95% CI = 1.25–4.66, P ≤ .008). Among patients, NL63 was associated with schizophrenia-spectrum (OR = 3.10, 95% CI = 1.27–7.58, P ≤ .013) but not mood disorders. HKU1 and NL63 coronavirus exposures may represent comorbid risk factors in neuropsychiatric disease. Future studies should explore links between the timing of coronavirus infections and subsequent development of schizophrenia and other disorders with psychotic symptoms.


title: Characteristics of respiratory virus infection during the outbreak of 2019 novel coronavirus in Beijing

text: Abstract Background Coronavirus disease 2019 (COVID-19) is spreading. Here, we summarized the composition of pathogens in fever clinic patients and analyzed characteristics of different respiratory virus infection. Methods Retrospectively collected patients with definite etiological results using nasal and pharyngeal swabs in fever clinic. Results Totally, 1860 patients were screened and 136 patients were enrolled. 72 (52.94%) of them were diagnosed as influenza (Flu) A virus infection. 32 (23.53%) of them were diagnosed as Flu B virus infection. 18 (13.24%) and 14 (10.29%) of them were diagnosed as COVID-19 and respiratory syncytial virus (RSV) infection respectively. COVID-19 group had a higher rate of contact with epidemic area within 14 days and clustering onset than other groups. Fever was the most common symptom in these patients. The ratio of fever and the highest temperature were higher in Flu A virus infection patients than in COVID-19 patients. COVID-19 patients had lower white blood cell count and neutrophil count than Flu A virus and RSV infection group, but higher lymphocyte count than Flu A and B virus infection groups. COVID-19 group (83.33%) had higher rate of pneumonia in chest CT scan than Flu A and B virus infection groups. Conclusions Influenza viruses accounted for a large proportion of respiratory virus infection even during the epidemic of COVID-19 in Beijing. No single symptom or laboratory finding was suggestive of specific respiratory virus, however, epidemic history was important for screening of COVID-19.


title: Timing of antiviral treatment initiation is critical to reduce SARS‐CoV‐2 viral load

text: We modeled the viral dynamics of 13 untreated patients infected with SARS‐CoV‐2 to infer viral growth parameters and predict the effects of antiviral treatments. In order to reduce peak viral load by more than 2 logs, drug efficacy needs to be greater than 90% if treatment is administered after symptom onset; an efficacy of 60% could be sufficient if treatment is initiated before symptom onset. Given their pharmacokinetic/pharmacodynamic properties, current investigated drugs may be in a range of 6‐87% efficacy. They may help control virus if administered very early, but may not have a major effect in severe patients.


title: Sequence analysis of nucleocapsid gene and leader RNA of human coronavirus OC43

text: Abstract The nucleotide sequence of the 3'-end of the genomic RNA of human coronavirus OC43 (HCV-OC43) was determined from the cDNA clones of the intracellular virus-specific mRNAs. The nucleotide sequence and the predicted amino acid sequence of the main open reading frame (ORF), which represents the nucleocapsid (N) protein, were highly homologous to those of bovine coronavirus (BCV) Mebus strain. This ORF predicts a protein of 448 amino acids. Additional smaller ORFs are also present in a different reading frame. We have also determined the leader sequence present at the 5'-end of HCV-OC43 mRNAs by a primer extension study. This sequence is highly homologous to that of mouse hepatitis virus, particularly in the 3'-end of the leader sequence, which is postulated to be involved in the unique mechanism of leader-primed transcription. These data suggest that HCV-OC43 and BCV might have diverged from each other fairly recently and that the 3'-end of the leader sequence has significant functional roles.


title: Severe Acute Respiratory Syndrome (SARS) and boarder protection: A report of the Sydney airport experience

text: Abstract In early 2003 Severe Acute Respiratory Syndrome (SARS) became an international globe-travelling virus; it began this frequent flyer journey from mainland China, and travelled to 30 countries across the world. By the end of its worldwide journey it had killed over 800 people and infected over 8000. As part of Australia's response to contain the virus, screening of all arriving international passengers was introduced throughout all of Australia's international airports. This article is a reflection on the experiences that five infection control practitioners (ICPs) had with implementing and conducting this service at Sydney's Mascot International Airport.


Critical care of Ebola patients: a crisis situation


title: Cellular 5′-3′ mRNA Exonuclease Xrn1 Controls Double-stranded RNA Accumulation and Anti-Viral Responses

text: By accelerating global mRNA decay, many viruses impair host protein synthesis, limiting host defenses and stimulating virus mRNA translation. Vaccinia virus (VacV) encodes two decapping enzymes (D9, D10) that remove protective 5′ caps on mRNAs, presumably generating substrates for degradation by the host exonuclease Xrn1. Surprisingly, we find VacV infection of Xrn1-depleted cells inhibits protein synthesis, compromising virus growth. These effects are aggravated by D9-deficiency and dependent upon a virus transcription factor required for intermediate and late mRNA biogenesis. Considerable double-stranded RNA (dsRNA) accumulation in Xrn1-depleted cells is accompanied by activation of host dsRNA-responsive defenses controlled by PKR and 2′-5′-oligoadenylate synthetase (OAS), which respectively inactivate the translation initiation factor eIF2 and stimulate RNA cleavage by RNase L. This proceeds despite VacV-encoded PKR and RNase L antagonists being present. Moreover, Xrn1-depletion sensitizes uninfected cells to dsRNA treatment. Thus, Xrn1 is a cellular factor regulating dsRNA accumulation and dsRNA-responsive innate immune effectors.


title: Virology 50th Anniversary Special Issue: A Publisher's Note

text: Abstract A Publishers Note introduces this special issue, commemorating Virology's 50th birthday, with a look back at its prestigious history, and the scientific progress that has been made in the field since the launch of the journal in 1954. It provides a list of the top 100 most-cited papers published in the journal to date.


title: Sample preparation for serum/plasma profiling and biomarker identification by mass spectrometry

text: Abstract In this article, we present an overview of the different strategies for sample preparation for identification by mass spectrometry (MS) of biomarkers from serum and/or plasma. We consider the effects of the variables involved in sample collection, handling and storage, and describe different approaches for removal of high abundance proteins and serum/plasma fractionation. We review the advantages and disadvantages of such techniques as centrifugal ultrafiltration, different formats for solid phase extraction, organic solvent extraction, gel and capillary electrophoresis, and liquid chromatography. We also discuss a variety of current proteomic methods and their main applications for biomarker-related studies.


The Cost of Koi: Evidence-Based Design in Emergency Medical Facilities