title: The role of epidermal growth factor receptor (EGFR) signaling in SARS coronavirus-induced pulmonary fibrosis

text: Abstract Many survivors of the 2003 outbreak of severe acute respiratory syndrome (SARS) developed residual pulmonary fibrosis with increased severity seen in older patients. Autopsies of patients that died from SARS also showed fibrosis to varying extents. Pulmonary fibrosis can be occasionally seen as a consequence to several respiratory viral infections but is much more common after a SARS coronavirus (SARS-CoV) infection. Given the threat of future outbreaks of severe coronavirus disease, including Middle East respiratory syndrome (MERS), it is important to understand the mechanisms responsible for pulmonary fibrosis, so as to support the development of therapeutic countermeasures and mitigate sequelae of infection. In this article, we summarize pulmonary fibrotic changes observed after a SARS-CoV infection, discuss the extent to which other respiratory viruses induce fibrosis, describe available animal models to study the development of SARS-CoV induced fibrosis and review evidence that pulmonary fibrosis is caused by a hyperactive host response to lung injury mediated by epidermal growth factor receptor (EGFR) signaling. We summarize work from our group and others indicating that inhibiting EGFR signaling may prevent an excessive fibrotic response to SARS-CoV and other respiratory viral infections and propose directions for future research.


title: Functional assessment and function‐based treatment delivered via telehealth: A brief summary

text: As the world navigates the COVID‐19 health crisis, behavior analysts are considering how best to support families while maintaining services and ensuring the health and safety of everyone involved. Telehealth is one service delivery option that provides families with access to care in their own communities and homes. In this article, we provide a brief summary of the telehealth literature in applied behavior analysis that provided coaching and training to families for individuals who displayed challenging behavior. These studies targeted functional assessment and function‐based treatment for challenging behavior. We briefly summarize what is known relative to the assessment and treatment of challenging behavior via telehealth, place these results within a descriptive context of the decisions made by the research team at the University of Iowa, and discuss what we, as behavior analysts, should consider next to advance our understanding and practice of telehealth.


DPP4-directed therapeutic strategies for MERS-CoV


title: The exposome in the community

text: The exposome is a complicated scientific research topic but an easily understandable concept for health. By integrating multiple external forces that act upon us, it is possible to take a holistic view of our health. The exposome model provides a way to organize and integrate the many new challenges we face in our day-to-day lives. This chapter provides examples of how the exposome can be incorporated into the lives of our families, communities, and countries, including educating the public across multiple sectors. The reader will find simple and scientifically sound recommendations for reducing chemical exposures and improving one’s exposome. Since one’s genome is relatively fixed, an improvement in one’s exposome will undoubtedly lead to an improvement in one’s health.


title: Loss of the Drosophila melanogaster DEAD box protein Ddx1 leads to reduced size and aberrant gametogenesis

text: Abstract Mammalian DDX1 has been implicated in RNA trafficking, DNA double-strand break repair and RNA processing; however, little is known about its role during animal development. Here, we report phenotypes associated with a null Ddx1 (Ddx1 AX ) mutation generated in Drosophila melanogaster. Ddx1 null flies are viable but significantly smaller than control and Ddx1 heterozygous flies. Female Ddx1 null flies have reduced fertility with egg chambers undergoing autophagy, whereas males are sterile due to disrupted spermatogenesis. Comparative RNA sequencing of control and Ddx1 null third instars identified several transcripts affected by Ddx1 inactivation. One of these, Sirup mRNA, was previously shown to be overexpressed under starvation conditions and implicated in mitochondrial function. We demonstrate that Sirup is a direct binding target of Ddx1 and that Sirup mRNA is differentially spliced in the presence or absence of Ddx1. Combining Ddx1 null mutation with Sirup dsRNA-mediated knock-down causes epistatic lethality not observed in either single mutant. Our data suggest a role for Drosophila Ddx1 in stress-induced regulation of splicing.


title: Bronchiectasis and cough: An old relationship in need of renewed attention

text: Abstract Bronchiectasis is an increasingly recognised respiratory condition with limited therapeutic options and a complex spectrum of clinical manifestations that invariably includes chronic cough. As the primary presentation of bronchiectasis in most cases, chronic cough and its mechanistic underpinnings are of central importance but remain poorly understood in this setting. Bronchiectasis is also increasingly identified as an underlying cause of chronic cough highlighting the interrelationship between the two conditions that share overlapping clinical features. Several therapeutic approaches have illustrated positive effects on bronchiectasis-associated cough, however, more focused treatment of heterogeneous cough subtypes may yield better outcomes for patients. A current challenge is the identification of bronchiectasis and cough endophenotypes that may allow improved patient stratification and more targeted therapeutic matching of the right treatment to the right patient. Here we discuss the complex disease phenotypes of bronchiectasis and their interrelationship with cough while considering current and emerging treatment options. We discuss some key cough promoters in bronchiectasis including infection, allergy and immune dysfunction.


title: F4-related mutation and expression analysis of the aminopeptidase N gene in pigs(1)

text: Intestinal infections with F4 enterotoxigenic Escherichia coli (ETEC) are worldwide an important cause of diarrhea in neonatal and recently weaned pigs. Adherence of F4 ETEC to the small intestine by binding to specific receptors is mediated by F4 fimbriae. Porcine aminopeptidase N (ANPEP) was recently identified as a new F4 receptor. In this study, 7 coding mutations and 1 mutation in the 3′ untranslated region (3' UTR)were identified in ANPEP by reverse transcriptase (RT–) PCR and sequencing using 3 F4 receptor-positive (F4R+) and 2 F4 receptor-negative (F4R–) pigs, which were F4 phenotyped based on the MUC4 TaqMan, oral immunization, and the in vitro villous adhesion assay. Three potential differential mutations (g.2615C > T, g.8214A > G, and g.16875C > G) identified by comparative analysis between the 3 F4R+ and 2 F4R– pigs were genotyped in 41 additional F4 phenotyped pigs. However, none of these 3 mutations could be associated with F4 ETEC susceptibility. In addition, the RT-PCR experiments did not reveal any differential expression or alternative splicing in the small intestine of F4R+ and F4R– pigs. In conclusion, we hypothesize that the difference in F4 binding to ANPEP is due to modifications in its carbohydrate moieties.


title: Porcine epidemic diarrhea virus infects and replicates in porcine alveolar macrophages

text: Abstract Porcine epidemic diarrhea virus (PEDV) is a causative agent of porcine epidemic diarrhea; consequently, the small intestine was believed to be its only target organ. In this study, we found that PEDV infected not only the small intestines, but also the respiratory tract. Infection and replication of PEDV in the respiratory tract from naturally PEDV-infected piglets were examined by reverse transcription polymerase chain reaction, immunohistochemistry, and virus re-isolation. Our observations were confirmed by experimental inoculation, and we found that PEDV infection in the respiratory tract was specifically associated with alveolar macrophages in the lung. Vero cell-adapted PEDV was able to replicate in both primary alveolar macrophages and continuous porcine alveolar macrophage cells. Sequencing analysis of the spike (S) glycoprotein revealed that mutations in S might be a potential determinant of auxiliary targets for PEDV. The discovery that PEDV infects and replicates in alveolar macrophages provides new insights into its pathogenesis.


title: Prevention of Respiratory Disease in Cow/Calf Operations

text: Respiratory disease in nursing calves is a common, yet sporadic herd event. In herds that experience respiratory disease, this can be a frustrating experience. Beef cow operations have an expectation of losses during the calving season. Losses because of respiratory disease in calves, that are at least 1 month up to weaning age, is not a normal expectation. Veterinarians providing services and advice to these operations must be able to present scientific and logical recommendations to manage these events. A strong working knowledge of the risk factors contributing to clinical disease is necessary to developing prevention strategies.


Coronavirus Disease 2019 (COVID-19) and men’s reproductive health


title: Future impacts and trends in treatment of hospital wastewater

text: The world’s population growth and economic development result in the increased requirement of land, water, and energy. This increased demand leads to the deforestation, loss in biodiversity, imbalance in agriculture and food supply, climate change, and increase in food and travel trade, which result in emergence and reemergence of infectious diseases. This chapter discussed various emerging infectious diseases and their causative agents (Buruli ulcer and Bunyvirus). Furthermore, this chapter further illustrates the emergence of superbugs and the associated threat due to the presence of pharmaceutical compounds in the environment. The prevalence of pharmaceuticals in the environment exerts ecotoxic effects on living organisms and causes thousands of death every year. The threats associated with the pharmaceutical presence in the environment were briefly discussed in this chapter. Finally, this chapter provides the alternative methods to avoid the use of antibiotics and to develop novel treatment technologies (such as Phage therapy) to degrade and remove the pharmaceutical compounds.


title: Current Evidence for Minimally Invasive Surgery During the COVID-19 Pandemic and Risk Mitigation Strategies: A Narrative Review

text: Our objective was to review the literature surrounding the risks of viral transmission during laparoscopic surgery and propose mitigation measures to address these risks. SUMMARY BACKGROUND DATA: The SARS-CoV-2 pandemic has caused surgeons the world over to re-evaluate their approach to surgical procedures given concerns over the risk of aerosolization of viral particles and exposure of operating room staff to infection. International society guidelines advise against the use of laparoscopy; however, the evidence on this topic is scant and recommendations are based on the perceived most cautious course of action. METHODS: We conducted a narrative review of the existing literature surrounding the risks of viral transmission during laparoscopic surgery and balance these risks against the benefits of minimally invasive approaches. We also propose mitigation measures to address these risks that we have adopted in our institution. RESULTS AND CONCLUSION: While it is currently assumed that open surgery minimizes operating room staff exposure to the virus, our findings reveal that this may not be the case. A well-informed, evidence-based opinion is critical when making decisions regarding which operative approach to pursue, for the safety and well-being of the patient, the operating room staff, and the healthcare system at large. Minimally invasive surgical approaches offer significant advantages with respect to both patient care, and the mitigation of the risk of viral transmission during surgery, provided the appropriate equipment and expertise are present.


Bleeding in COVID-19 severe pneumonia: the other side of abnormal coagulation pattern?


title: Avian reovirus sigma C enhances the mucosal and systemic immune responses elicited by antigen-conjugated lactic acid bacteria

text: Abstract Mucosal surfaces are common sites of pathogen colonization/entry. Effective mucosal immunity by vaccination should provide protection at this primary infection site. Our aim was to develop a new vaccination strategy that elicits a mucosal immune response. A new strain of Enterococcus faecium, a non pathogenic lactic acid bacteria (LAB) with strong cell adhesion ability, was identified and used as a vaccine vector to deliver two model antigens. Specifically, sigma (σ) C protein of avian reovirus (ARV), a functional homolog of mammalian reovirus σ1 protein and responsible for M-cell targeting, was administered together with a subfragment of the spike protein of infectious bronchitis virus (IBV). Next, the effect of immunization route on the immune response was assessed by delivering the antigens via the LAB strain. Intranasal (IN) immunization induced stronger humoral responses than intragastic (IG) immunization. IN immunization produced antigen specific IgA both systemically and in the lungs. A higher IgA titer was induced by the LAB with ARV σC protein attached. Moreover, the serum of mice immunized with LAB displaying divalent antigens had much stronger immune reactivity against ARV σC protein compared to IBV-S1. Our results indicate that ARV σC protein delivered by LAB via the IN route elicits strong mucosal immunity. A needle-free delivery approach is a convenient and cost effective method of vaccine administration, especially for respiratory infections in economic animals. Furthermore, ARV σC, a strong immunogen of ARV, may be able to serve as an immunoenhancer for other vaccines, especially avian vaccines.


title: Surveillance to improve evidence for community control decisions during the COVID-19 pandemic – Opening the animal epidemic toolbox for public health

text: Abstract During the first few months of 2020, the COVID-19 pandemic has reached Europe. Health systems all over the world are trying to control the outbreak in the shortest possible time. Exotic disease outbreaks are not uncommon in animal health and randomised surveillance is frequently used as support for decision-making. This editorial discusses the possibilities of practicing One Health, by using methods from animal health to enhance surveillance for COVID-19 to provide an evidence base fort decision-making in communities and countries.


title: S-palmitoylation of swine interferon-inducible transmembrane protein is essential for its anti-JEV activity

text: Japanese encephalitis virus (JEV) is an infectious pathogen spreading in a wide range of vertebrate species. Pigs are amplifying hosts of JEV and thought to be maintained in nature predominantly by avian-mosquito cycles. In the innate immune system, interferon-inducible transmembrane protein (IFITM) is a small transmembrane protein family and has been identified as the first line of defense against a broad range of RNA virus invasion. In this paper, we found that swine IFITM (sIFITM) could restrict the replication of both JEV vaccine strain and wild strain NJ-2008. The cysteine S-palmitoylation modification of sIFITM plays important roles in their anti-JEV effects and intracellular distributions. Our findings show the anti-JEV activities of swine interferon-inducible transmembrane proteins and broaden the antiviral spectrum of IFITM protein family. The preliminary exploration of S-palmitoylation modification of sIFITM may contribute to understanding of the antiviral molecular mechanism of sIFITM.


title: Middle East respiratory syndrome coronavirus (MERS-CoV) — Surveillance and testing in North England from 2012 to 2019

text: Abstract Background Middle East respiratory syndrome coronavirus (MERS-CoV) emerged in Saudi Arabia in 2012 and caused an epidemic in the Middle East. Public Health England (PHE) Manchester is one of the two PHE centres in the UK that perform testing for MERS-CoV. The results of the PHE Manchester MERS surveillance from 2012 to 2019 are presented in this report. Methods Retrospective data were collected for returning travellers from the Middle East fitting the PHE MERS case definition. Respiratory samples were tested for respiratory viruses and MERS-CoV using an in-house RT-PCR assay. Results Four hundred and twenty-six (426) samples from 264 patients were tested for MERS Co-V and respiratory viruses. No MERS-CoV infections were identified by PCR. Fifty-six percent of samples were PCR positive for viral or bacterial pathogen with Influenza A as the predominant virus (44%). Sixty-two percent of all patients had a pathogen identified with the highest positivity from sputum samples. Patients with multiple samples demonstrated a 100% diagnostic yield. Conclusions Although no cases of MERS were identified, the majority of patients had Influenza infection for which oseltamivir treatment was indicated and isolation warranted. Sputum samples were the most useful in diagnosing respiratory viruses with a 100% diagnostic yield from patients with multiple samples.


title: Evaluation of the Temporal Association between Kawasaki Disease and Viral Infections in South Korea

text: BACKGROUND AND OBJECTIVES: This study is aimed at elucidating potential temporal associations between the occurrence of Kawasaki disease (KD) and various viral infections. SUBJECTS AND METHODS: We obtained monthly patterns of KD from the seventh nationwide survey and viral detection data from the Korea Centers for Disease Control and Prevention from 2009 to 2011 and evaluated temporal correlations between them for each month. The respiratory viruses detected using a multiplex real-time-polymerase chain reaction kit were influenza virus (A/H1N1, A/H3N2, A/H5N1, and B), adenovirus, parainfluenza virus (type 1, 2, 3), respiratory syncytial virus (type A, B), human rhinovirus, human coronavirus (OC43/229E, NL63), human bocavirus, and enterovirus. RESULTS: We obtained data from a total of 13031 patients who were treated for acute KD from 87 hospitals with pediatric residence programs. During this survey, KD showed highest overall incidence in summer and winter seasons and lowest incidence in February and October. We received viral detection data for a total of 14267 patients. Viral detection was highest during winter and spring seasons. The most commonly detected virus was human rhinovirus (32.6%), followed by influenza virus (26.8%). The monthly incidence of KD showed significant correlation with the monthly overall viral detection (p=0.022, r=0.382). In particular, human bocavirus and enterovirus have significant correlations with monthly patterns of KD occurrence (p=0.032 and p=0.007, respectively) and influenza virus correlated with KD occurrence with borderline significance (p=0.063). CONCLUSION: The temporal association between monthly occurrence of KD and viral detection suggests the etiologic importance of precedent infection in the development of KD.


title: Pediatric Airway Management in Coronavirus Disease 2019 Patients: Consensus Guidelines From the Society for Pediatric Anesthesia’s Pediatric Difficult Intubation Collaborative and the Canadian Pediatric Anesthesia Society

text: The severe acute respiratory syndrome Coronavirus 2 (Coronavirus Disease 2019 [COVID-19]) pandemic has challenged medical systems and clinicians globally to unforeseen levels. Rapid spread of COVID-19 has forced clinicians to care for patients with a highly contagious disease without evidence-based guidelines. Using a virtual modified nominal group technique, the Pediatric Difficult Intubation Collaborative (PeDI-C), which currently includes 35 hospitals from 6 countries, generated consensus guidelines on airway management in pediatric anesthesia based on expert opinion and early data about the disease. PeDI-C identified overarching goals during care, including minimizing aerosolized respiratory secretions, minimizing the number of clinicians in contact with a patient, and recognizing that undiagnosed asymptomatic patients may shed the virus and infect health care workers. Recommendations include administering anxiolytic medications, intravenous anesthetic inductions, tracheal intubation using video laryngoscopes and cuffed tracheal tubes, use of in-line suction catheters, and modifying workflow to recover patients from anesthesia in the operating room. Importantly, PeDI-C recommends that anesthesiologists consider using appropriate personal protective equipment when performing aerosol-generating medical procedures in asymptomatic children, in addition to known or suspected children with COVID-19. Airway procedures should be done in negative pressure rooms when available. Adequate time should be allowed for operating room cleaning and air filtration between surgical cases. Research using rigorous study designs is urgently needed to inform safe practices during the COVID-19 pandemic. Until further information is available, PeDI-C advises that clinicians consider these guidelines to enhance the safety of health care workers during airway management when performing aerosol-generating medical procedures. These guidelines have been endorsed by the Society for Pediatric Anesthesia and the Canadian Pediatric Anesthesia Society.


title: Systemic infection of avian influenza A virus H5N1 subtype in humans

text: Summary The viral dissemination in a patient with avian influenza A subtype H5N1 infection was retrospectively studied by the immunohistochemical localization of viral nucleoprotein antigen. The pathology was marked by diffuse alveolar damage, lymphoid depletion, and reactive hemophagocytic syndrome. Besides the lung and the upper respiratory tract, viral antigen was detected in the small and large intestinal epithelial cells, hematopoietic cells in the bone marrow, glial cells and neurons of the brain, and lymphocytes. The results confirmed that H5N1 virus disseminated to multiple organs beyond the respiratory system. However, specific pathological changes were noted in the respiratory system only, and productive viral replication confirmed by culture was noted only in the lung. More postmortem studies are needed to elucidate the pathogenesis of this highly fatal zoonotic disease.


Comment on “Hearing loss and COVID-19: A note”


title: SEIR Modeling of the Italian Epidemic of SARS-CoV-2 Using Computational Swarm Intelligence

text: We applied a generalized SEIR epidemiological model to the recent SARS-CoV-2 outbreak in the world, with a focus on Italy and its Lombardy, Piedmont, and Veneto regions. We focused on the application of a stochastic approach in fitting the model parameters using a Particle Swarm Optimization (PSO) solver, to improve the reliability of predictions in the medium term (30 days). We analyzed the official data and the predicted evolution of the epidemic in the Italian regions, and we compared the results with the data and predictions of Spain and South Korea. We linked the model equations to the changes in people’s mobility, with reference to Google’s COVID-19 Community Mobility Reports. We discussed the effectiveness of policies taken by different regions and countries and how they have an impact on past and future infection scenarios.


title: Atypical erythema multiforme palmar plaques lesions due to Sars‐Cov‐2

text: Since CoronaVirus Disease 19 (COVID-19) was declared as a pandemic, all medical specialties were at the front, including dermatology. Daily, there are observations mentioning possible cutaneous manifestations of Sars-Cov-2. Among these manifestations, the most often was a rash which could be erythematous, morbilliform or urticarial mimicking viral exanthema, or chickenpox-like vesicles [1-3]. Anecdotally, dengue-like petechial eruption was reported [4].


title: Sex Differences in Proatherogenic Cytokine Levels

text: Background: It has been shown that sex affects immunity, including cytokine production. Given that atherosclerosis is an inflammatory disease promoted by specific cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, we aimed at evaluating whether sex could affect the levels of these proatherogenic cytokines in a group of healthy adults. In this analysis, we also included other cytokines and peptides that have been implicated in atherosclerosis development and progression. Methods: A total of 104 healthy adults were recruited; we measured circulating levels of IL-1β, IL-6, TNF-α, angiotensins and angiotensin-converting enzyme-2 (ACE2), as well as osteoprotegerin and receptor activator of nuclear factor κB ligand (RANKL). Results: IL-1β, IL-6, and TNF-α were significantly higher in men as compared to women. They were all associated with testosterone and the testosterone/estradiol ratio. They remained significantly associated with sex (but not with hormones) after being tested for potential confounders. Conclusions: Sex seems to influence the levels of proatherogenic cytokines. This is consistent not only with sex differences in vulnerability to infections but also with the higher cardiovascular risk exhibited by the male gender as compared to the female gender. Nevertheless, this association is only partly explained by hormone levels.


title: Optimizing Medication Treatment of Opioid Use Disorder During COVID-19 (SARS-CoV-2)

text: The COVID-19 health crisis joined, rather than supplanted, the opioid crisis as the most acutely pressing threats to US public health. In the setting of COVID-19, opioid use disorder treatment paradigms are being disrupted, including the fact that methadone clinics are scrambling to give “take-home” doses where they would typically not. The rapid transition away from in-person examination, dosing and group therapy in an era of social isolation calls for adjustments to clinical practice, including emphasizing patient-provider communication, favoring new inductees on buprenorphine and leveraging technology to optimize safety of medication treatment.


title: Analytical evaluation of the microarray-based FluChip-8G Influenza A+B Assay

text: Abstract Background Influenza causes a significant annual disease burden, with characterization of the infecting virus important in clinical and public health settings. Rapid immunoassays are fast but insensitive, whereas real-time RT-PCR is sensitive but susceptible to genetic mutations and often requires multiple serial assays. The FluChip-8G Influenza A+B Assay provides type and subtype/lineage identification of influenza A and B, including non-seasonal A viruses, in a single microarray-based assay with same day turnaround time. Objective To evaluate key analytical performance characteristics of the FluChip-8G Influenza A+B Assay. Study design Analytical sensitivity, cross-reactivity, and multi-site reproducibility were evaluated. Results The limit of detection (LOD) for the FluChip-8G influenza A+B Assay ranged from 5.8 × 102–1.5 × 105 genome copies/mL, with most samples ∼2 × 103 genome copies/mL (∼160 genome copies/reaction). Fifty two (52) additional strains were correctly identified near the LOD, demonstrating robust reactivity. Two variant viruses (H1N1v and H3N2v) resulted in dual identification as both “non-seasonal influenza A” and A/H1N1pdm09. No reproducible cross-reactivity was observed for the 34 organisms tested, however, challenges with internal control inhibition due to crude growth matrix were observed. Lastly, samples tested near the LOD showed high reproducibility (97.0% (95% CI 94.7–98.7)) regardless of operator, site, reagent lot, or testing day. Conclusion The FluChip-8G Influenza A+B Assay is an effective new method for detecting and identifying both seasonal and non-seasonal influenza viruses, as revealed by good sensitivity and robust reactivity to 52 unique strains of influenza virus. In addition, the lack of cross-reactivity to non-influenza pathogens and high lab-to-lab reproducibility highlight the analytical performance of the assay as an alternative to real-time RT-PCR and sequencing-based assays. Clinical validation of the technology in a multi-site clinical study is the subject of a separate investigation.


title: The multifaceted exosome: Biogenesis, role in normal and aberrant cellular function, and frontiers for pharmacological and biomarker opportunities

text: Abstract Exosomes are bioactive vesicles derived from the cell's endosomal membrane system and secreted into surrounding body fluids. Exosomes contain cell and cell-state specific cargos of protein, mRNA and miRNA. Exosome formation, cargo content, and delivery to surrounding cells is of immense biological interest considering the role that exosomes are believed to play in various pathological conditions. They aid antigen presentation by immune cells and can exhibit either anti-inflammatory or pro-inflammatory properties depending on the parent antigen-presenting cell's conditioning. Viruses can hijack a host cell's exosomal machinery to evade host defense systems aiding in the trans-infection of viruses. Tumor derived exosomes may help establish an oncogenic niche systemically via delivery of protein, mRNA, and miRNA that can aid angiogenesis, cell proliferation, and cell survival. Exosomes have also been implicated in the spread of neurodegenerative diseases. Studies have shown that exosomes are selectively taken up by cells distal from their release. They can reprogram the recipient cells due to their active molecular cargo. Cell-lineage and state-specific exosomes imply that they may therefore harbor body fluid-based biomarkers of unparalleled accuracy, particularly for tissues that are difficult or impossible to access. Exosome-specific membrane proteins provide markers enabling exosome identity and selection, while cell type and cell condition-specific protein, mRNA and miRNA cargo provide a rich potential source of biomarkers. This review serves to provide an overview of the current state of the science in the burgeoning field of exosome biology.


title: Respiratory macrophages regulate CD4 T memory responses to mucosal immunization with recombinant adenovirus-based vaccines

text: Abstract Respiratory immunization is an attractive way to generate systemic and mucosal protective memory responses that are required for preventing mucosally transmitted infections. However, the molecular and cellular mechanisms for controlling memory T cell responses remain incompletely understood. In this study, we investigated the role of respiratory macrophage (MΦ) in regulating CD4 T cell responses to recombinant adenovirus-based (rAd) vaccines. We demonstrated that rAd intranasal (i.n.) vaccination induced migration and accumulation of respiratory MΦ and circulatory monocytes in the mediastinal lymph nodes and lung parenchyma. Under the influence of respiratory MΦ CD4 T cells exhibited slow proliferation kinetics and an increased tendency of generating central memory, as opposed to effector memory, CD4 T cell responses in vitro and in vivo. Correspondingly, depletion of MΦ using clodronate-containing liposome prior to i.n. immunization significantly enhanced CD4 T cell proliferation and increased the frequency of CD4 memory T cells in the airway lumen, demonstrating that MΦ initially serve as a negative regulator in limiting generation of mucosal tissue-resident memory CD4 T cells. However, clodronate-containing liposome delivery following i.n. immunization markedly reduced the frequencies of memory CD4 T cells in the airway lumen and spleen, indicating that respiratory MΦ and potentially circulating monocytes are critically required for maintaining long-term memory CD4 T cells. Collectively, our data demonstrate that rAd-induced mucosal CD4 T memory responses are regulated by respiratory MΦ and/or monocytes at multiple stages.


title: Analysis of human P[4]G2 rotavirus strains isolated in Brazil reveals codon usage bias and strong compositional constraints

text: Abstract The Rotavirus genus belongs to the family Reoviridae and its genome consist of 11 segments of double-stranded RNA. Group A rotaviruses (RV-A) are the main etiological agent of acute viral gastroenteritis in infants and young children worldwide. Understanding the extent and causes of biases in codon usage is essential to the understanding of viral evolution. However, the factors shaping synonymous codon usage bias and nucleotide composition in human RV-A are currently unknown. In order to gain insight into these matters, we analyzed the codon usage and base composition constraints on the two genes that codify the two outer capsid proteins (VP4 [VP8*] and VP7) of 58 P[4]G2 RV-A strains isolated in Brazil and investigated the possible key evolutionary determinants of codon usage bias. The results of these studies revealed that the frequencies of codon usage in both RV-A proteins studied are significantly different than the ones used by human cells. In order to observe if similar trends of codon usage are found when RV-A complete genomes are considered, we compare these results with results found using a dataset of 10 reference strains for whom the complete codes of the 11 segments are known. Similar results were obtained using capsid proteins or complete genomes. The general correlations found between the position of each sequence on the first axis generated by correspondence analysis and the relative dinucleotide abundances indicate that codon usage in RV-A can also be strongly influenced by underlying biases in dinucleotide frequencies. CpG and GpC containing codons are markedly suppressed. Thus, the results of this study suggest that RV-A genomic biases are the result of the evolution of genome composition in relation to host adaptation and the ability to escape antiviral cell responses.


title: Why Aging T Cells Fail: Implications for Vaccination

text: Summary The decline in CD4+ T cell function with aging contributes to reduced vaccine efficacy. In this commentary, we discuss the factors leading to age-related changes in T cell function and propose how they may be overcome to enhance vaccine efficacy for the elderly.


title: Lower respiratory tract infection in the community: associations between viral aetiology and illness course

text: Abstract Objectives This study determined associations between respiratory viruses and subsequent illness course in primary care adult patients presenting with acute cough and/or suspected lower respiratory tract infection (LRTI). Methods A prospective European primary care study recruited adults with symptoms of lower respiratory tract infection between Nov-Apr 2007-2010. Real-time in-house polymerase chain reaction (PCR) was performed to test for six common respiratory viruses. In this secondary analysis, symptom severity (scored 1=no problem, 2=mild, 3=moderate, 4=severe) and symptom duration were compared between groups with different viral aetiologies using regression and Cox proportional hazard models, respectively. Additionally, associations between baseline viral load (cycle threshold (Ct) value) and illness course were assessed. Results The PCR tested positive for a common respiratory virus in 1,354 of the 2,957 (45.8%) included patients. The overall mean symptom score at presentation was 2.09 (95%CI 2.07-2.11) and the median duration until resolution of moderately bad or severe symptoms was 8.70 days (interquartile range 4.50-11.00). Patients with influenza virus, human metapneumovirus (hMPV), respiratory syncytial virus (RSV), coronavirus (CoV) or rhinovirus had a significantly higher symptom score than patients with no virus isolated (0.07-0.25 points or 2.3-8.3% higher symptom score). Time to symptom resolution was longer in RSV infections (adjusted hazard ratio (AHR) 0.80, 95%CI 0.65-0.96) and hMPV infections (AHR 0.77, 95%CI 0.62-0.94) than in infections with no virus isolated. Overall, baseline viral load was associated with symptom severity (difference 0.11, 95%CI 0.06-0.16 per 10 cycles decrease in Ct value), but not with symptom duration. Conclusions In healthy, working adults from the general community presenting at the general practitioner with acute cough and/or suspected LRTI respiratory viruses other than influenza impose an illness burden comparable to influenza. Hence, the public health focus for viral respiratory tract infections should be broadened.


title: A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents

text: Abstract Medical and veterinary diagnostic and public health laboratories world-wide are increasingly being called upon to introduce molecular diagnostic tests for both endemic and exotic diseases. This demand has accelerated following increasing terrorism fears. Ironically these same concerns have lead to tightening of both import and export controls preventing many laboratories, particularly those outside of the United States, from gaining access to positive control material. This in turn has prevented many laboratories from introducing much needed molecular diagnostic tests. We describe here a generic approach for preparing synthetic DNA or RNA control material for use in either TaqMan or conventional PCR assays. The production of synthetic controls using this approach does not require cloning or special equipment or facilities beyond that found in any laboratory performing molecular diagnostics. The approach significantly reduces the possibility of contamination or erroneously reporting false-positive reactions due to contamination from positive control material. Synthetic controls produced using this approach have been employed in all molecular diagnostic tests performed in our laboratory and are used irrespective of whether we possess the organism or not.


title: Venipuncture in bats

text: Though not as common as small rodents in laboratory settings, bats are being increasingly used in research studies. Knowledge of proper blood sampling techniques is essential for care and management of bats.


title: Clinical Trials and Tribulations in the COVID-19 Era

text: Advances in treating and preventing Alzheimer's disease and other neurocognitive disorders of aging arise from rigorous preclinical and clinical research, with randomized controlled treatment trials as the last and definitive test. The COVID-19 pandemic has greatly disrupted ongoing interventional studies and researchers are scrambling to find ways to safely continue this critical work amidst rapidly shifting guidelines from sponsors, institutions, and state and federal guidelines. Here we describe novel approaches and work-flow adaptations to study visits, drug delivery and interim and endpoint safety and outcomes assessments to avoid sacrificing years of preparation and substantial financial investments, to work in the best interest of participants and their caregivers, and to continue on the path towards discovering disease-modifying treatments for the millions of individuals impacted by major neurocognitive disorders.


Estimation of COVID-19 burden in Egypt – Authors' reply


title: A Functional Henipavirus Envelope Glycoprotein Pseudotyped Lentivirus Assay System

text: BACKGROUND: Hendra virus (HeV) and Nipah virus (NiV) are newly emerged zoonotic paramyxoviruses discovered during outbreaks in Queensland, Australia in 1994 and peninsular Malaysia in 1998/9 respectively and classified within the new Henipavirus genus. Both viruses can infect a broad range of mammalian species causing severe and often-lethal disease in humans and animals, and repeated outbreaks continue to occur. Extensive laboratory studies on the host cell infection stage of HeV and NiV and the roles of their envelope glycoproteins have been hampered by their highly pathogenic nature and restriction to biosafety level-4 (BSL-4) containment. To circumvent this problem, we have developed a henipavirus envelope glycoprotein pseudotyped lentivirus assay system using either a luciferase gene or green fluorescent protein (GFP) gene encoding human immunodeficiency virus type-1 (HIV-1) genome in conjunction with the HeV and NiV fusion (F) and attachment (G) glycoproteins. RESULTS: Functional retrovirus particles pseudotyped with henipavirus F and G glycoproteins displayed proper target cell tropism and entry and infection was dependent on the presence of the HeV and NiV receptors ephrinB2 or B3 on target cells. The functional specificity of the assay was confirmed by the lack of reporter-gene signals when particles bearing either only the F or only G glycoprotein were prepared and assayed. Virus entry could be specifically blocked when infection was carried out in the presence of a fusion inhibiting C-terminal heptad (HR-2) peptide, a well-characterized, cross-reactive, neutralizing human mAb specific for the henipavirus G glycoprotein, and soluble ephrinB2 and B3 receptors. In addition, the utility of the assay was also demonstrated by an examination of the influence of the cytoplasmic tail of F in its fusion activity and incorporation into pseudotyped virus particles by generating and testing a panel of truncation mutants of NiV and HeV F. CONCLUSIONS: Together, these results demonstrate that a specific henipavirus entry assay has been developed using NiV or HeV F and G glycoprotein pseudotyped reporter-gene encoding retrovirus particles. This assay can be conducted safely under BSL-2 conditions and will be a useful tool for measuring henipavirus entry and studying F and G glycoprotein function in the context of virus entry, as well as in assaying and characterizing neutralizing antibodies and virus entry inhibitors.


title: Porcine deltacoronavirus infection: Etiology, cell culture for virus isolation and propagation, molecular epidemiology and pathogenesis

text: Abstract Porcine deltacoronavirus (PDCoV) (family Coronaviridae, genus Deltacoronavirus) is a novel swine enteropathogenic coronavirus that causes acute diarrhea/vomiting, dehydration and mortality in seronegative neonatal piglets. PDCoV diarrhea was first reported in the US in early 2014, concurrently with co-circulation of porcine epidemic diarrhea virus (PEDV) (family Coronaviridae, genus Alphacoronavirus). The origin of PDCoV in pigs and also its sudden emergence or route of introduction into the US still remains unclear. In the US, since 2013–2014, the newly emerged PDCoV and PEDV have spread nationwide, causing a high number of pig deaths and significant economic impacts. The current US PDCoV strains are enteropathogenic and infect villous epithelial cells of the entire small and large intestines although the jejunum and ileum are the primary sites of infection. Similar to PEDV infections, PDCoV infections also cause acute, severe atrophic enteritis accompanied by transient viremia (viral RNA) that leads to severe diarrhea and/or vomiting, followed by dehydration as the potential cause of death in nursing piglets. At present, differential diagnosis of PDCoV, PEDV, and transmissible gastroenteritis virus (TGEV) is essential to control viral diarrheas in US swine. Cell culture-adapted US PDCoV (TC-PDCoV) strains have been isolated and propagated by us and in several other laboratories. TC-PDCoV strains will be useful to develop serologic assays and to evaluate if serial cell-culture passage attenuates TC-PDCoV as a potential vaccine candidate strain. A comprehensive understanding of the pathogenesis and epidemiology of epidemic PDCoV strains is currently needed to prevent and control the disease in affected regions and to develop an effective vaccine. This review focuses on the etiology, cell culture isolation and propagation, molecular epidemiology, disease mechanisms and pathogenesis of PDCoV infection.


title: Viral and bacterial coinfection among hospitalized children with respiratory tract infections

text: BACKGROUND: The epidemiology of Mycoplasma pneumoniae (MP) and local dominant etiologies of pathogens that cause respiratory tract infections (RTIs) among central China children (≤14 years old) hospitalized are poorly understood. METHODS: A total of 10,429 specimens were analyzed, and IgM antibodies against 9 respiratory pathogens including MP were detected using indirect immunofluorescence assay from serum. RESULTS: It showed that 59.3% of the enrolled children were positive for at least 1 pathogen; highest detection rates included those between 3 and <6 years of age (70.4%), female (63.2%), and who were hospitalized in 2014 (80.9%). The most predominant pathogen was MP (45.6%), followed by Parainfluenza viruses (PIVs) (22.6%) and influenza B viruses (IFVB) (14.7%). Coinfection was observed in 2,907 specimens (27.9%); the coinfection combination containing MP and PIVs had the highest detection rate of 15%, followed by MP and IFVB as well as IFVB and PIVs. CONCLUSIONS: MP was the most commonly detected bacteria among hospitalized children, which should be included in the differential diagnosis for hospitalized children with RTI. These findings will contribute to the effective prevention and therapeutic approaches of pathogens among local children suffering from RTI.


title: Gene therapy in The Netherlands: highlights from the Low Countries

text: Gene therapy is an active research area in The Netherlands and Dutch scientists involved in fundamental and clinical gene therapy research significantly contribute to the progresses made in this field. This ranges from the establishment of the 293, 911 and PER.C6 cell lines, which are used worldwide for the production of replication‐defective adenoviral vectors, to the development of targeted viral vectors and T lymphocytes as well as of non‐viral vectors. Several milestones have been achieved in Dutch clinical gene therapy trials, including the first treatment worldwide of patients with adenosine deaminase deficiency with genetically corrected hematopoietic stem cells in collaboration with French and British scientists. Until now, about 230 patients with various diseases have been treated with viral and non‐viral gene therapy in this country. Ongoing and upcoming Dutch clinical trials focus on the translation of new developments in gene therapy research, including the restoration of genetic defects other than SCID, and the use of oncolytic adenoviruses and targeted T cells for the treatment of cancer. The growing commercial interest in Dutch clinical gene therapy is reflected by the involvement of two Dutch companies in ongoing trials as well as the participation of Dutch clinical centres in large phase III international multicenter immuno‐gene therapy trials on prostate cancer sponsored by an American company. Translational gene therapy research in The Netherlands is boosted at a governmental level by the Dutch Ministry of Health via a dedicated funding programme. This paper presents an overview on milestones in Dutch basic gene therapy research as well as on past, present and future clinical gene therapy trials in The Netherlands. Copyright © 2007 John Wiley & Sons, Ltd.


title: Digital epidemiology and global health security; an interdisciplinary conversation

text: Contemporary infectious disease surveillance systems aim to employ the speed and scope of big data in an attempt to provide global health security. Both shifts - the perception of health problems through the framework of global health security and the corresponding technological approaches – imply epistemological changes, methodological ambivalences as well as manifold societal effects. Bringing current findings from social sciences and public health praxis into a dialogue, this conversation style contribution points out several broader implications of changing disease surveillance. The conversation covers epidemiological issues such as the shift from expert knowledge to algorithmic knowledge, the securitization of global health, and the construction of new kinds of threats. Those developments are detailed and discussed in their impacts for health provision in a broader sense.


title: Statistical Analysis of Forecasting COVID-19 for Upcoming Month in Pakistan

text: In this paper, we have conducted analysis based on data obtained from National Institute of Health (NIH) - Islamabad and produced a forecast of COVID-19 confirmed cases as well as the number of deaths and recoveries in Pakistan using the Auto-Regressive Integrated Moving Average Model (ARIMA). The fitted forecasting models revealed high exponential growth in the number of confirmed cases, deaths and recoveries in Pakistan. Based on our model prediction the number of confirmed cases will be increased by 2.7 times, 95% prediction interval for the number of cases at the end of May 2020 = (5681 to 33079). There could be up to 500 deaths, 95% prediction interval = (168 to 885) and there could be eightfold increase in the number of recoveries, 95% prediction interval = (2391 to 16126). The forecasting results of COVID-19 are alarming for May in Pakistan. The health officials and government should adopt new strategies to control the pandemic from further spread until a proper treatment or vaccine is developed.


title: The association between Ostertagia ostertagi antibodies in bulk tank milk samples and parameters linked to cattle reproduction and mortality

text: Abstract In Western Europe, gastrointestinal nematodes are widespread in dairy cattle. This study was carried out to evaluate the relationship between optical density ratio (ODR) measured on bulk tank milk with an indirect Ostertagia ostertagi ELISA and reproduction/mortality parameters. Data were collected between 2008 and 2010 from monitoring carried out on 1643 dairy herds (Normandy, Western France). ODR values of 3 samples from each farm taken from November 2008 to 2010 were averaged and then transformed into a categorical variable. Reproductive and mortality data were obtained from 1444 herds using cow records from government databases. Statistical analysis was carried out using ordinary logistic regression (OLR). The outcome variables were the case–control status of a herd for reproductive factors, age at first calving and inter-calving intervals, and mortality ratios of various age classes. The effect of the categorical ODR variable was studied and several potential confounder herd factors were used to improve the model fit. A significant relationship was found between high Ostertagia ODR levels and a late age at the first calving (>34.5 months) (odds ratio (OR)=1.94, p <0.001). No significant relationship was observed with OLR for inter-calving intervals although bivariate analysis showed that herds with high ODR levels had longer inter-calving intervals than herds with low ODR level (first inter-calving interval in herds with low vs. high ODR levels=412 days vs. 422 days, p <0.001; other inter-calving intervals=408 days vs. 413 days, p <0.01). A high ODR level was also associated with high mortality of calves between 0 and 30 days of life (mortality ratio>6%) (OR=1.43, p <0.05) and between 91 and 365 days (ratio>3%) (OR=1.72, p <0.01). No significant relationship was observed with multivariate approach for mortalities in other classes by age, but bivariate analysis showed that herds with high ODR level had higher mortalities than herds with low ODR levels (mortality between 31 and 90 days in herds with low vs. high ODR levels=1.89% vs. 2.91%, p <0.001; mortality after 365 days=1.67% vs. 2.93%, p <0.001). In conclusion, our results confirm the usefulness of ELISA as an indicator for production losses in dairy herds. This inexpensive tool could be advantageous, used to aid farmers and veterinarians to carry out appropriate control measures.


Presumed Asymptomatic Carrier Transmission of COVID-19.


title: Pulsed electromagnetic fields stimulation prevents steroid-induced osteonecrosis in rats

text: BACKGROUND: Pulsed electromagnetic fields (PEMF) stimulation has been used successfully to treat nonunion fractures and femoral head osteonecrosis, but relatively little is known about its effects on preventing steroid-induced osteonecrosis. The purpose of the study was to investigate the effects of PEMF stimulation on the prevention of steroid-induced osteonecrosis in rats and explore the underlying mechanisms. METHODS: Seventy-two male adult Wistar rats were divided into three groups and treated as follows. (1) PEMF stimulation group (PEMF group, n = 24): intravenously injected with lipopolysaccharide (LPS, 10 μg/kg) on day 0 and intramuscularly injected with methylprednisolone acetate (MPSL, 20 mg/kg) on days 1, 2 and 3, then subjected to PEMF stimulation 4 h per day for 1 to 8 weeks. (2) Methylprednisolone-treated group (MPSL group, n = 24): injected the same dose of LPS and MPSL as the PEMF group but without exposure to PEMF. (3) Control group (PS group, n = 24): injected 0.9% saline in the same mode at the same time points. The incidence of osteonecrosis, serum lipid levels and the mRNA and protein expression of transforming growth factor β1 (TGF-β1) in the proximal femur were measured 1, 2, 4 and 8 weeks after the last MPSL (or saline) injection. RESULTS: The incidence of osteonecrosis in the PEMF group (29%) was significantly lower than that observed in the MPSL group (75%), while no osteonecrosis was observed in the PS group. The serum lipid levels were significantly lower in the PEMF and PS groups than in the MPSL group. Compared with the MPSL and PS groups, the mRNA expression of TGF-β1 increased, reaching a peak 1 week after PEMF treatment, and remained high for 4 weeks, then declined at 8 weeks, whereas the protein expression of TGF-β1 increased, reaching a peak at 2 weeks after PEMF treatment, and remained high for 8 weeks. CONCLUSIONS: PEMF stimulation can prevent steroid-induced osteonecrosis in rats, and the underlying mechanisms involve decreased serum lipid levels and increased expression of TGF-β1.


title: Diagnosis of Small Intestinal Disorders in Dogs and Cats

text: Laboratory tests are an important part of the workup of small intestinal diseases in dogs and cats. Especially in chronic cases, when extragastrointestinal causes need to be ruled out, it is important to adhere to a systematic workup. This article details the newest available data on tests to aid this diagnostic process. Once the diagnosis of a chronic enteropathy is made, there are many laboratory tests that can help in monitoring the disease and providing prognostic information. Several new tests being evaluated for clinical usefulness are discussed.


title: MERS-CoV diagnosis: An update

text: Summary Diagnosis of MERS-Cov still a major concern in most of daignostic laboratories. To date the Real-time Polymerase Chain reaction (RT-PCR) is the mainstay for diagnosis of MERS-CoV. RT-PCR has limitations, including a long turnaround time and lack of common measurements and correlations with Viral Load (VL). It is recommended to screen for MERS-CoV using RT-PCR of the upstream of envelope gene (upE) followed by confirmation of the presence of one of the following genes; open reading frame 1A, 1B genes or nucleocapsid (N) gene. Scientists are looking to implement viral sequencing on all negative samples by RT-PCR and they beleive that can be exposed to another level of testing using sequencing of the RNA-dependent RNA polymerase (RdRp) gene or N gene and in this case a positive result is diagnostic. It is also very important to maintain a contineous and random sequencing for MERS-Cov samples to be able to pick early viral mutations. Serological assays still not widely or routinely performed, and a lot of studies looking to implement such method in routine patient's testings.


1092. Adenovirus-Based Vaccine Prevents Pneumonia in Ferrets Challenged with the SARS Coronavirus


Challenges and Opportunities for Lung Ultrasound in Novel Coronavirus Disease (COVID-19).


Liberté, égalité, fraternité à l’épreuve du COVID-19


title: Adenovirus-based vaccine prevents pneumonia in ferrets challenged with the SARS coronavirus and stimulates robust immune responses in macaques

text: A ferret model of severe acute respiratory syndrome (SARS)-CoV infection was used to evaluate the efficacy of an adenovirus vaccine. Animals were subjected to heterologous prime-boost using vectors from human serotype 5 and chimpanzee derived adenoviruses (human AdHu5 and chimpanzee AdC7) expressing spike protein followed by intranasal challenge with SARS-CoV. Vaccination led to a substantial reduction in viral load and prevented the severe pneumonia seen in unvaccinated animals. The same prime-boost strategy was effective in rhesus macaques in eliciting SARS-CoV specific immune responses. These data indicate that a heterologous adenovirus-based prime-boost vaccine strategy could safely stimulate strong immunity that may be needed for complete protection against SARS-CoV infection.


title: Comparative studies of the capsid precursor polypeptide P1 and the capsid protein VP1 cDNA vectors for DNA vaccination against foot‐and‐mouth disease virus

text: BACKGROUND: Foot‐and‐mouth disease virus (FMDV) causes a severe livestock disease, and the virus is an interesting target for virology and vaccine studies. MATERIALS AND METHODS: Here we evaluated comparatively three different viral antigen‐encoding DNA sequences, delivered via two physical means (i.e., gene gun delivery into skin and electroporation delivery into muscle), for naked DNA‐mediated vaccination in a mouse system. RESULTS: Both methods gave similar results, demonstrating commonality of the observed DNA vaccine effects. Immunization with a cDNA vector expressing the major viral antigen (VP1) alone routinely failed to induce the production of anti‐VP1 or neutralizing antibodies in test mice. As a second approach, the plasmid L‐VP1 that produces a transgenic membrane‐anchored VP1 protein elicited a strong antibody response, but all test mice failed in the FMDV challenge experiment. In contrast, for mice immunized with the viral capsid precursor protein (P1) cDNA expression vector, both neutralizing antibodies and 80–100% protection in test mice were detected. CONCLUSIONS: This strategy of using the whole capsid precursor protein P1 cDNA for vaccination, intentionally without the use of virus‐specific protease or other encoding genes for safety reasons, may thus be employed as a relevant experimental system for induction or upgrading of effective neutralizing antibody response, and as a convenient surrogate test system for DNA vaccination studies of FMDV and presumably other viral diseases. Copyright © 2005 John Wiley & Sons, Ltd.


title: Immunomodulatory Activity and Protective Effects of Polysaccharide from Eupatorium adenophorum Leaf Extract on Highly Pathogenic H5N1 Influenza Infection

text: The development of novel broad-spectrum, antiviral agents against H5N1 infection is urgently needed. In this study, we evaluated the immunomodulatory activities and protective effect of Eupatorium adenophorum polysaccharide (EAP) against the highly pathogenic H5N1 subtype influenza virus. EAP treatment significantly increased the production of IL-6, TNF-α, and IFN-γ both in vivo and in vitro as measured by qPCR and ELISA. In a mouse infection model, intranasal administration of EAP at a dose of 25 mg/kg body weight prior to H5N1 viral challenge efficiently inhibited viral replication, decreased lung lesions, and increased survival rate. We further evaluated the innate immune recognition of EAP, as this process is regulated primarily Dectin-1 and mannose receptor (MR). These results indicate that EAP may have immunomodulatory properties and a potential prophylactic effect against H5N1 influenza infection. Our investigation suggests an alternative strategy for the development of novel antiinfluenza agents and benefits of E. adenophorum products.


title: Identification of critical airports for controlling global infectious disease outbreaks: Stress-tests focusing in Europe

text: Abstract As the global population increases and transportation connectivity improves in quality and prices, the demand for mobility increases, especially in long-haul services. According to the 2017 report of the European Commission in Mobility and Transport, the performance of all modes for passenger transport (roadways and airways) are reaching record highs. Although the benefits of the increased demand for mobility are substantial and welcome, an effort should be paid such as to ameliorate possible threatening side-effects that may also arise. As World Health Organization (WHO) denotes and as has been evident from the global COVID-19 epidemic outbreak, infectious diseases can be spread directly or indirectly from one person to another under common exposure circumstances such as air transportation (especially long-haul airline connections) that may act as the medium for transmitting and spreading infectious diseases. In this paper, analytical and realistic models have been integrated, for providing evidence on the spread dynamics of infectious diseases that may face Europe through the airlines system. In particular, a detailed epidemiological model has been integrated with the airlines’ and land transport network, able to simulate the epidemic spread of infectious diseases originated from distant locations. Additionally, a wide set of experiments and simulations have been conducted, providing results from detailed stress-tests covering both mild as well as aggressive cases of epidemic spreading scenarios. The results provide convincing evidence on the effectiveness that the European airports' system offer in controlling the emergence of epidemics, but also on the time and extent that controlling measures should be taken in order to break the chain of infections in realistic cases.


title: Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties

text: Abstract Twenty 5-fluorouracil-induced temperature-sensitive (ts) mutants of mouse hepatitis virus strain A59 were isolated from 1284 virus clones. Mutants were preselected on the basis of their inability to induce syncytia in infected cells at the restrictive temperature (40°) vs the permissive temperature (31°). Of these mutants, only those with a relative plating efficiency 40° 31° of 3 × 10−3 or smaller were kept. Virus yields at 40° compared to 37° and 31° (leakiness) were determined. Most mutants (16) were RNA−, i.e., unable to synthesize virus-specific RNA at the restrictive temperature. The other four were RNA+. No qualitative differences were detected in the virus-specific RNAs in cells infected with RNA+ ts-mutants, both at 31° and 40°. Virus-specific proteins present in cells infected with is-171 (RNA−) and the RNA+-mutants (ts-43, is-201, is-209, and is-279) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitates. No qualitative differences in the pattern of virus-specific cellular proteins were detected among the mutants except for an additional polypeptide of about 46,000 daltons in ts-209-infected cells. Finally, the neuropathogenic properties of eight of the mutants were investigated. Whereas 102 PFU of wild-type virus injected intracerebrally killed 50 to 100% of 4-week-old Balc/c mice within 1 week, the mutants were highly attenuated. A dose of 105 PFU lead to no or transient disease. However, 4 weeks after infection with ts 342, is-43, or is-201 obvious histological changes were observed in brain and spinal cord of clinically healthy mice.


title: Preparation and characterization of polyoxometalate-modified poly(vinyl alcohol)/polyethyleneimine hybrids as a chemical and biological self-detoxifying material

text: Abstract Nanohybrid membranes based on the Keggin-type polyoxometalate (POM) H5PV2Mo10O40 and a poly(vinyl alcohol)/polyethyleneimine (PVA/PEI) blend were prepared as a chemical and biological protective material. The objective of the study was to develop and evaluate permeable membranes (PVA/PEI) impregnated with reactive nanoparticulates (POM) that can protect against simulants of chemical and biological warfare agents. The physical properties of the PVA/PEI–POM hybrids were examined using SEM, TEM, TGA, and UV–Vis spectroscopy, the results of which indicated that the POM was incorporated in the PVA/PEI matrix after impregnation. The redox properties against 2-chloroethyl-ethyl sulfide (CEES) were investigated based on significant color changes and UV absorption in the POM upon reduction by CEES. The antibacterial effects of the PVA/PEI–POM hybrids were assessed by the zone of inhibition, minimum inhibitory concentration (MIC), and plate-counting methods. The results of this study showed that PVA/PEI–POM hybrids that act against simulants of chemical and biological weapons while retaining their ability to transmit moisture vapor could be obtained.


title: COVID-19: From bench to bed side

text: Abstract Background and aims The last two decades have experienced the outbreaks of three different coronaviruses in the different parts of the world namely; Severe acute respiratory syndrome cornonavirus-1 (SARS-CoV-1), Middle East respiratory syndrome (MERS-CoV) and Severe acute respiratory syndrome cornonavirus-2 (SARS-CoV-2). We aimed to delineate the differences in viral dynamics and clinical features between them and tried to focus on every basic details of SARS-COV-2 (COVID-19) that every health care provider must know. Methods We systematically searched the PubMed database up till April 2, 2020 and retrieved all the articles published on SARS-CoV-2, SARS-CoV-1, MERS-CoV that dealt with viral dynamics. Results Ample data is available to suggest the differences in etiology, transmission cycle, diagnosis, genetics, hosts, reproductive rates, clinical features, laboratory diagnosis and radiological features between SARS-CoV-1, MERS-CoV and SARS-CoV-2. Conclusion Although SARS-CoV-2 (COVID-19) is more infectious than SARS-CoV-1 and MERS-CoV, most infections are generally mild and self-limiting. However, case-fatality rates are very high in patients with COVID-19 with comorbidities, compared to SARS-CoV-1 and MERS-CoV.


title: Vanadium. Its Role for Humans

text: Vanadium is the 21st most abundant element in the Earth’s crust and the 2nd-to-most abundant transition metal in sea water. The element is ubiquitous also in freshwater and nutrients. The average body load of a human individual amounts to 1 mg. The omnipresence of vanadium hampers checks directed towards its essentiality. However, since vanadate can be considered a close blueprint of phosphate with respect to its built-up, vanadate likely takes over a regulatory function in metabolic processes depending on phosphate. At common concentrations, vanadium is non-toxic. The main source for potentially toxic effects caused by vanadium is exposure to high loads of vanadium oxides in the breathing air of vanadium processing industrial enterprises. Vanadium can enter the body via the lungs or, more commonly, the stomach. Most of the dietary vanadium is excreted. The amount of vanadium resorbed in the gastrointestinal tract is a function of its oxidation state (V(V) or V(IV)) and the coordination environment. Vanadium compounds that enter the blood stream are subjected to speciation. The predominant vanadium species in blood are vanadate and vanadyl bound to transferrin. From the blood stream, vanadium becomes distributed to the body tissues and bones. Bones act as storage pool for vanadate. The aqueous chemistry of vanadium(V) at concentration <10 μM is dominated by vanadate. At higher concentrations, oligovanadates come in, decavanadate in particular, which is thermodynamically stable in the pH range 2.3–6.3, and can further be stabilized at higher pH by interaction with proteins. The similarity between vanadate and phosphate accounts for the interplay between vanadate and phosphate-dependent enzymes: phosphatases can be inhibited, kinases activated. As far as medicinal applications of vanadium compounds are concerned, vanadium’s mode of action appears to be related to the phosphate-vanadate antagonism, to the direct interaction of vanadium compounds or fragments thereof with DNA, and to vanadium’s contribution to a balanced tissue level of reactive oxygen species. So far vanadium compounds have not yet found approval for medicinal applications. The antidiabetic (insulin-enhancing) effect, however, of a singular vanadium complex, bis(ethylmaltolato)oxidovanadium(IV) (BEOV), has revealed encouraging results in phase IIa clinical tests. In addition, in vitro studies with cell cultures and parasites, as well as in vivo studies with animals, have revealed a broad potential spectrum for the application of vanadium coordination compounds in the treatment of cardiac and neuronal disorders, malignant tumors, viral and bacterial infections (such as influenza, HIV, and tuberculosis), and tropical diseases caused by parasites, e.g., Chagas’ disease, leishmaniasis, and amoebiasis.


title: Multiplex PCR and Microarray for Detection of Swine Respiratory Pathogens

text: Porcine respiratory disease complex (PRDC) is one of the most important health concerns for pig producers and can involve multiple viral and bacterial pathogens. No simple, single‐reaction diagnostic test currently exists for the simultaneous detection of major pathogens commonly associated with PRDC. Furthermore, the detection of most of the bacterial pathogens implicated in PRDC currently requires time‐consuming culture‐based methods that can take several days to obtain results. In this study, a novel prototype automated microarray that integrates and automates all steps of post‐PCR microarray processing for the simultaneous detection and typing of eight bacteria and viruses commonly associated with PRDC is described along with associated multiplex reverse transcriptase PCR. The user‐friendly assay detected and differentiated between four viruses [porcine reproductive and respiratory syndrome virus (PRRSV), influenza A virus, porcine circovirus type 2, porcine respiratory corona virus], four bacteria (Mycoplasma hyopneumoniae, Pasteurella multocida, Salmonella enterica serovar Choleraesuis, Streptococcus suis), and further differentiated between type 1 and type 2 PRRSV as well as toxigenic and non‐toxigenic P. multocida. The assay accurately identified and typed a panel of 34 strains representing the eight targeted pathogens and was negative when tested with 34 relevant and/or closely related non‐target bacterial and viral species. All targets were also identified singly or in combination in a panel of clinical lung samples and/or experimentally inoculated biological material.


title: Deep Sequencing for the Detection of Virus-Like Sequences in the Brains of Patients with Multiple Sclerosis: Detection of GBV-C in Human Brain

text: Multiple sclerosis (MS) is a demyelinating disease of unknown origin that affects the central nervous system of an estimated 400,000 Americans. GBV-C or hepatitis G is a flavivirus that is found in the serum of 1–2% of blood donors. It was originally associated with hepatitis, but is now believed to be a relatively non-pathogenic lymphotropic virus. Fifty frozen specimens from the brains of deceased persons affected by MS were obtained along with 15 normal control brain specimens. RNA was extracted and ribosomal RNAs were depleted before sequencing on the Illumina GAII. These 36 bp reads were compared with a non-redundant database derived from the 600,000+ viral sequences in GenBank organized into 4080 taxa. An individual read successfully aligned to the viral database was considered to be a “hit”. Normalized MS specimen hit rates for each viral taxon were compared to the distribution of hits in the normal controls. Seventeen MS and 11 control brain extracts were sequenced, yielding 4–10 million sequences (“reads”) each. Over-representation of sequence from at least one of 12 viral taxa was observed in 7 of the 17 MS samples. Sequences resembling other viruses previously implicated in the pathogenesis of MS were not significantly enriched in any of the diseased brain specimens. Sequences from GB virus C (GBV-C), a flavivirus not previously isolated from brain, were enriched in one of the MS samples. GBV-C in this brain specimen was confirmed by specific amplification in this single MS brain specimen, but not in the 30 other MS brain samples available. The entire 9.4 kb sequence of this GBV-C isolate is reported here. This study shows the feasibility of deep sequencing for the detection of occult viral infections in the brains of deceased persons with MS. The first isolation of GBV-C from human brain is reported here.


title: 5.42 RNA Interference Technology

text: This chapter replaces Z. Liang, P. Zhang, 5.15 - RNA Interference (RNAi) Technology, Editor: Murray Moo-Young, Comprehensive Biotechnology (Second Edition), Academic Press, 2011, Pages 179-187.


title: CORRELATION BETWEEN CHEST COMPUTED TOMOGRAPHY AND LUNG ULTRASONOGRAPHY IN PATIENTS WITH CORONAVIRUS DISEASE 2019 (COVID-19)

text: There is growing evidence regarding the imaging findings of Coronavirus Disease 2019 (COVID-19), in chest X-ray and Computed Tomography scan (CT). At this moment, the role of Lung Ultrasonography (LUS) has yet to be explored. The main purpose of this study is to evaluate the correlation between LUS findings and chest CT in confirmed (positive RT-PCR) or clinically highly suspicious (dyspnea, fever, myasthenia, gastrointestinal symptoms, dry cough, ageusia or anosmia) of COVID-19 patients. This prospective study was carried out in the emergency department (ED), confirmed or clinically highly suspicious COVID-19 patients were recruited and were subjected to a chest CT and concurrent LUS exam. An experienced ED physician performed the LUS exam blind to the clinical history and results of the CT scan, which were reviewed by two radiologists in consensus for signs compatible with COVID-19 (bilateral ground-glass opacities in peripheral distribution). Compatible LUS exam was considered a bilateral pattern of B-lines, irregular pleural line and subpleural consolidations. Between March and April 2020, fifty-one patients were consecutively enrolled. The indication for CT was a negative or indeterminate RT-PCR test (49.0%) followed by suspicion of pulmonary embolism (41.2%). Radiological signs compatible with COVID-19 were present in thirty-seven patients (72.5%) on CT scan and forty patients (78.4%) on LUS exam. The presence of LUS findings was correlated with a positive CT scan suggestive of COVID-19 (OR: 13.3, 95%CI: 4.5-39.6, p<0.001) with a sensitivity of 100.0% and a specificity of 78.6%, positive predictive value of 92.5% and negative of 100.0%. There was no missed diagnosis of COVID-19 with LUS compared to CT in our cohort. The LUS Score had a good correlation with CT total severity score (ICC 0.803, 95% CI 0.60-0.90, p<0.001). LUS presents similar accuracy compared to chest CT to detect lung abnormalities in COVID-19 patients.


title: COHORT OF 4404 PERSONS UNDER INVESTIGATION FOR COVID-19 IN A NY HOSPITAL AND PREDICTORS OF ICU CARE AND VENTILATION

text: ABSTRACT Study objective Most COVID-19 reports have focused on SARS-CoV-2 positive patients. However, at the time of initial presentation, most patients’ viral status is unknown. Determination of factors that predict initial and subsequent need for intensive care (ICU) and invasive mechanical ventilation (IMV) are critical for resource planning and allocation. We describe our experience with 4,404 persons under investigation (PUI) and explore predictors of ICU care and IMV at a NY COVID-19 epicenter. Methods We conducted a retrospective COHORT of all persons under investigation (PUI) presenting to a large academic medical center emergency department (ED) in NYS with symptoms suggestive of COVID-19. The association between patient predictor variables and SARS-CoV-2 status, intensive care unit (ICU) admission, invasive mechanical ventilation (IMV), and mortality were explored with univariate and multivariate analyses. Results Between March 12-April 14, 2020 we saw 4,404 PUI patients of whom 68% were discharged home, 29% were admitted to a regular floor and 3% to an ICU. 1,651 of 3,369 patients tested were SARS-CoV-2 positive to date. Of regular floor admits, 13% were subsequently upgraded to the ICU after a median (IQR) of 62 (28-106) hrs. 50 patients required IMV in the ED, 4 required prehospital IMV, and another 167 subsequently required IMV in a median (IQR) of 60 (26-99) hours after admission. Testing positive for SARS-CoV-2 and lower oxygen saturations were associated with need for ICU, IMV and death. High respiratory rates were associated with the need for ICU care. Conclusions PUI for COVID-19 contribute significantly to the healthcare burden beyond those ruling in for SARS-CoV-2. For every 100 admitted PUI, 9 will require ICU and/or IMV upon arrival and another 12 within 2-3 days of hospital admission, especially PUIs with lower oxygen saturations and positive SARS-CoV-2 swabs. This information should help hospitals stay ahead of the pandemic curve.


title: Discovery and development of safe-in-man broad-spectrum antiviral agents

text: Abstract Viral diseases are one of the leading causes of morbidity and mortality in the world. Virus-specific vaccines and antiviral drugs are the most powerful tools to combat viral diseases. However, broad-spectrum antiviral agents (BSAAs, i.e. compounds targeting viruses belonging to two or more viral families) could provide additional protection of the general population from emerging and re-emerging viral diseases, reinforcing the arsenal of available antiviral options. Here, we review discovery and development of BSAAs and summarize the information on 120 safe-in-man agents in a freely accessible database (https://drugvirus.info/). Future and ongoing pre-clinical and clinical studies will increase the number of BSAAs, expand the spectrum of their indications, and identify drug combinations for treatment of emerging and re-emerging viral infections as well as co-infections.


title: Two known therapies could be useful as adjuvant therapy in critical patients infected by COVID-19

text: Abstract Pneumonia caused by coronavirus, which originated in Wuhan, China, in late 2019, has been spread around the world already becoming a pandemic. Unfortunately, there is not yet a specific vaccine or effective antiviral drug for treating COVID-19. Many of these patients deteriorate rapidly and require intubation and are mechanically ventilated, which is causing the collapse of the health system in many countries due to lack of ventilators and intensive care beds. In this document we review two simple adjuvant therapies to administer, without side effects, and low cost that could be useful for the treatment of acute severe coronavirus infection associated with acute respiratory syndrome (SARS-CoV-2). VitaminC, a potent antioxidant, has emerged as a relevant therapy due to its potential benefits when administered intravenous. The potential effect of vitaminC in reducing inflammation in the lungs could play a key role in lung injury caused by coronavirus infection. Another potential effective therapy is ozone: it has been extensively studied and used for many years and its effectiveness has been demonstrated so far in multiples studies. Nevertheless, our goal is not to make an exhaustive review of these therapies but spread the beneficial effects themselves. Obviously clinical trials are necessaries, but due to the potential benefit of these two therapies we highly recommended to add to the therapeutic arsenal.


title: Qualitative study of a quarantine/isolation model with multiple disease stages

text: Abstract Recent studies suggest that, for disease transmission models with latent and infectious periods, the use of gamma distribution assumption seems to provide a better fit for the associated epidemiological data in comparison to the use of exponential distribution assumption. The objective of this study is to carry out a rigorous mathematical analysis of a communicable disease transmission model with quarantine (of latent cases) and isolation (of symptomatic cases), in which the waiting periods in the infected classes are assumed to have gamma distributions. Rigorous analysis of the model reveals that it has a globally-asymptotically stable disease-free equilibrium whenever its associated reproduction number is less than unity. The model has a unique endemic equilibrium when the threshold quantity exceeds unity. The endemic equilibrium is shown to be locally and globally-asymptotically stable for special cases. Numerical simulations, using data related to the 2003 SARS outbreaks, show that the cumulative number of disease-related mortality increases with increasing number of disease stages. Furthermore, the cumulative number of new cases is higher if the asymptomatic period is distributed such that most of the period is spent in the early stages of the asymptomatic compartments in comparison to the cases where the average time period is equally distributed among the associated stages or if most of the time period is spent in the later (final) stages of the asymptomatic compartments. Finally, it is shown that distributing the average sojourn time in the infectious (asymptomatic) classes equally or unequally does not effect the cumulative number of new cases.


title: Diagnostic performance of two highly multiplexed respiratory virus assays in a pediatric cohort

text: Abstract Background Rapid detection of respiratory viruses is important for management and infection control in hospitalized patients. Multiplex nucleic acid tests (NATs) have begun to replace conventional methods as gold standards for respiratory virus detection. Objective To compare the performance of two large multiplex NATS, ResPlex II (RPII) and Respiratory Virus Surveillance kit with electrospray ionization mass spectrometry (RVS/MS) using nasopharyngeal aspirates (NPAs) from hospitalized children who had been tested previously with conventional methods. Study design Stored residual NPAs (N =306) were tested concomitantly by RPII and RVS/MS. Alternate NATs were used to adjudicate discordant results. Results More viruses were detected with multiplex NATs (RPII, 110; RVS/MS, 109) than conventional assays (86); diagnostic gain was primarily for fastidious viruses (coronaviruses and enteroviruses [EVs]/human rhinoviruses [HRVs]). Total positive and negative agreement between the multiplex NATs for all viruses detected was quite high (86% positive agreement, 99% negative agreement). Most individual viruses were detected with fairly equivalent accuracy by the multiplex NATs, except for adenoviruses (RPII sensitivity 40%) and human metapneumovirus (RVS/MS sensitivity 42%). RPII had the advantage of detecting EVs and HRVs, however, it demonstrated considerable EV/HRV cross-reactivity (29 HRV-positive specimens by real-time PCR were positive for EV by RPII and 21 specimens positive for HRV only by RT-PCR were dual positive for EV/HRV by RPII). RPII also had reduced sensitivity for HRV detection (in 36 specimens, HRV was detected by RT-PCR but not by RPII). Conclusions Both multiplex NATs were promising, but had notable limitations.


title: Cross host transmission in the emergence of MERS coronavirus

text: Coronaviruses (CoVs) able to infect humans emerge through cross-host transmission from animals. There is substantial evidence that the recent Middle East respiratory syndrome (MERS)-CoV outbreak is fueled by zoonotic transmission from dromedary camels. This is largely based on the fact that closely related viruses have been isolated from this but not any other animal species. Given the widespread geographical distribution of dromedaries found seropositive for MERS-CoV, continued transmission may likely occur in the future. Therefore, a further understanding of the cross host transmission of MERS-CoV is needed to limit the risks this virus poses to man.


title: Connecting viral with cellular interactomes

text: Genome-scale screens for intraviral and virus–host protein interactions and the analysis of literature-curated datasets are able to provide a novel, comprehensive perspective of viruses, and virus-infected cells. Until now, large-scale interaction screens were predominantly performed with the yeast-two-hybrid (Y2H) system; however, alternative high-throughput technologies detecting binary protein interactions or protein complexes have been developed. Although many of the previous studies suffer from a rather poor validation of the results and few biological implications, these technologies potentially lead to a plethora of novel hypotheses. Here, we will give an overview of current approaches and their technical limitations, present recent examples and novel developments.


Introduction: emerging importance of the rhinovirus


US health agencies take stock after federal shutdown


title: Risk factors for feline coronavirus seropositivity in cats relinquished to a UK rescue charity

text: Abstract Two thousand, two hundred and seven cats from 14 shelters of a major UK cat charity were blood tested for feline coronavirus (FCoV) antibodies. Data was collated on breed, sex, age, number of cats at original location, outdoor access, health status, and time spent in the shelter prior to sampling (range 0 to 4 years). Some cats were also tested for feline leukaemia virus antigen, feline immunodeficiency virus, and Toxoplasma gondii antibodies. The effect of these variables on FCoV seropositivity was explored by multivariable logistic regression. FCoV seropositivity in cats that had spent 5 days or less in a shelter at sampling was significantly associated with a multi-cat origin, cats aged 3 years or less, and Persian breed. Whether pet, stray or feral, health status, indoor/outdoor access, and sex had no significant effect. Overall FCoV seropositivity was associated with time spent in a shelter but this association was not linear. Cats that had spent more than 60 days in a shelter were over five times as likely to be seropositive. This may be the result of a change in husbandry from solitary to communal housing for cats remaining in shelters long term. Rescue of cats for less than 60 days was not associated with a significant increasing risk of seropositivity. Significant variation existed in seropositivity between individual shelters overall and in cats rescued for less than 5 days. These findings may reflect inter-shelter variation in cat husbandry and variation in seropositivity of shelter intake respectively.


title: Field Disease Diagnostic Investigation of Neonatal Calf Diarrhea

text: Diarrhea is a leading cause of sickness and death of beef and dairy calves in their first month of life. Field investigations of outbreaks of neonatal calf diarrhea should be conducted for the purposes of (1) reducing the losses associated with existing cases and (2) preventing new cases from occurring. The appropriate corrective actions are determined by taking a population approach to diagnostics and prevention. It can be difficult to provide solutions to disease outbreaks, but success is more likely if an organized, epidemiologic approach to outbreak investigation is followed.


title: Porcine Torovirus (PToV)—A Brief Review of Etiology, Diagnostic Assays and Current Epidemiology

text: Porcine torovirus (PToV) is a potential enteric swine pathogen, found at especially high rates in piglets with diarrhea. It was first reported in the Netherlands in 1998 and has emerged in many countries around the world. Infections are generally asymptomatic and have not directly caused large economic losses, though co-infections with other swine pathogens and intertype recombination may lead to unpredictable outcomes. This review introduces progress in PToV research regarding its discovery, relationship with other Toroviruses, virion morphological characteristics, genetic structure and variation, recent epidemiology, diagnostic methods, and possibilities for future research.


title: Polymorphisms of human leucocyte antigen genes in Maonan people in China

text: We examined human leucocyte antigen (HLA) gene polymorphisms in the Maonan people from southern China. HLA‐A, ‐B and ‐DRB1 alleles were determined in 108 healthy unrelated Maonan individuals by the polymerase chain reaction‐Luminex method, and haplotype frequencies for HLA‐A, ‐B and ‐DRB1 loci were estimated. The most frequent HLA‐A alleles were A*1101 (35.2%), A*0203 (17.6%), A*0207 (13.4%) and A*2402 (13.4%); HLA‐B alleles were B*1301(19.9%), B*1502 (14.8%), B*4601 (13.4%) and B*4001 (13.4%); HLA‐DRB1 alleles were DRB1*1202 (17.1%), DRB1*1602 (13.0%) and DRB1*1401 (10.7%). The most common haplotypes were A*0207‐B*4601 (10.6%), A*1101‐B*1301 (10.0%), A*1101‐B*4001 (8.4%), B*1502‐DRB1*1202 (12.0%), B*4601‐DRB1*1401 (5.8%), A*1101‐B*1502‐DRB1*1202 (7.1%) and A*0207‐B*4601‐DRB1*1401 (5.3%), profiles that are also found in populations from the southern region of East Asia. Phylogenetic and principal component analyses revealed that the Maonan people belong to the southeastern Asian group and are most closely related to the Buyi people.


title: A novel dynamic flow immunochromatographic test (DFICT) using gold nanoparticles for the serological detection of Toxoplasma gondii infection in dogs and cats

text: A novel dynamic flow immunochromatographic test (DFICT) is proposed for rapid assay utilizing Toxoplasma gondii as a model. The test is based on a proprietary technology that combines the principles of immunochromatography and fluid dynamics. Gold nanoparticles conjugated to staphylococcal protein A (SPA) were prepared in liquid form and used as signal vehicles. T. gondii-specific recombinant antigens and SPA were sprayed onto a nitrocellulose membrane in strips at positions designated as T and C, respectively. The DFICT is performed by applying a 100 µL aliquot of liquid gold-SPA conjugate to the reagent hole and a 5 μL aliquot of serum sample to the sample hole. The results were observable within 5 min by the naked eye. The lowest detectable limit of the assay was determined as the highest dilution (1:320) of positive serum. No cross-reaction of the antibodies with other related canine or feline pathogens was observed. The DFICT can be stored for 12 months at 4 °C or 6 months with no loss of sensitivity or specificity. A high degree of consistency was observed between the DFICT and the standard ELISA kit, supporting the reliability of the novel test strip. The introduction of a liquid gold nanoparticle conjugate reagent provides this method with several attractive characteristics, such as ease of manufacture, low sample volume requirements, high selectivity and high efficiency. This method opens a novel pathway for rapid diagnostic screening and field analysis.


title: Caveolin-1 and -2 in airway epithelium: expression and in situ association as detected by FRET-CLSM

text: BACKGROUND: Caveolae are involved in diverse cellular functions such as signal transduction, cholesterol homeostasis, endo- and transcytosis, and also may serve as entry sites for microorganisms. Hence, their occurrence in epithelium of the airways might be expected but, nonetheless, has not yet been examined. METHODS: Western blotting, real-time quantitative PCR analysis of abraded tracheal epithelium and laser-assisted microdissection combined with subsequent mRNA analysis were used to examine the expression of cav-1 and cav-2, two major caveolar coat proteins, in rat tracheal epithelium. Fluorescence immunohistochemistry was performed to locate caveolae and cav-1 and -2 in the airway epithelium of rats, mice and humans. Electron-microscopic analysis was used for the identification of caveolae. CLSM-FRET analysis determined the interaction of cav-1α and cav-2 in situ. RESULTS: Western blotting and laser-assisted microdissection identified protein and transcripts, respectively, of cav-1 and cav-2 in airway epithelium. Real-time quantitative RT-PCR analysis of abraded tracheal epithelium revealed a higher expression of cav-2 than of cav-1. Immunoreactivities for cav-1 and for cav-2 were co-localized in the cell membrane of the basal cells and basolaterally in the ciliated epithelial cells of large airways of rat and human. However, no labeling for cav-1 or cav-2 was observed in the epithelial cells of small bronchi. Using conventional double-labeling indirect immunofluorescence combined with CLSM-FRET analysis, we detected an association of cav-1α and -2 in epithelial cells. The presence of caveolae was confirmed by electron microscopy. In contrast to human and rat, cav-1-immunoreactivity and caveolae were confined to basal cells in mice. Epithelial caveolae were absent in cav-1-deficient mice, implicating a requirement of this caveolar protein in epithelial caveolae formation. CONCLUSION: These results show that caveolae and caveolins are integral membrane components in basal and ciliated epithelial cells, indicating a crucial role in these cell types. In addition to their physiological role, they may be involved in airway infection.


title: Heparan Sulfate-Dependent Enhancement of Henipavirus Infection

text: Nipah virus and Hendra virus are emerging, highly pathogenic, zoonotic paramyxoviruses that belong to the genus Henipavirus. They infect humans as well as numerous mammalian species. Both viruses use ephrin-B2 and -B3 as cell entry receptors, and following initial entry into an organism, they are capable of rapid spread throughout the host. We have previously reported that Nipah virus can use another attachment receptor, different from its entry receptors, to bind to nonpermissive circulating leukocytes, thereby promoting viral dissemination within the host. Here, this attachment molecule was identified as heparan sulfate for both Nipah virus and Hendra virus. Cells devoid of heparan sulfate were not able to mediate henipavirus trans-infection and showed reduced permissivity to infection. Virus pseudotyped with Nipah virus glycoproteins bound heparan sulfate and heparin but no other glycosaminoglycans in a surface plasmon resonance assay. Furthermore, heparin was able to inhibit the interaction of the viruses with the heparan sulfate and to block cell-mediated trans-infection of henipaviruses. Moreover, heparin was shown to bind to ephrin-B3 and to restrain infection of permissive cells in vitro. Consequently, treatment with heparin devoid of anticoagulant activity improved the survival of Nipah virus-infected hamsters. Altogether, these results reveal heparan sulfate as a new attachment receptor for henipaviruses and as a potential therapeutic target for the development of novel approaches against these highly lethal infections.


title: Ring Vaccination and Smallpox Control

text: We present a stochastic model for the spread of smallpox after a small number of index cases are introduced into a susceptible population. The model describes a branching process for the spread of the infection and the effects of intervention measures. We discuss scenarios in which ring vaccination of direct contacts of infected persons is sufficient to contain an epidemic. Ring vaccination can be successful if infectious cases are rapidly diagnosed. However, because of the inherent stochastic nature of epidemic outbreaks, both the size and duration of contained outbreaks are highly variable. Intervention requirements depend on the basic reproduction number R(0), for which different estimates exist. When faced with the decision of whether to rely on ring vaccination, the public health community should be aware that an epidemic might take time to subside even for an eventually successful intervention strategy.


Multidisciplinary guidance to manage comatose patients with severe COVID‐19


Perceived Challenges of COVID-19 Infection Prevention and Control Preparedness: A Multinational Survey


title: Localization of the virus neutralizing and hemagglutinin epitopes of E1 glycoprotein of rubella virus

text: Abstract Current serological assays using whole rubella virus (RV) as a target antigen for detecting RV-specific antibodies fail to define specific RV proteins and antigenic determinants such as hemagglutinin (HA) and virus-neutralizing (VN) epitopes of rubella virus. A panel of El deletion mutants and a subset of E1-specific monoclonal antibodies (MAb) were used for the initial analysis of HA and VN epitopes of E1 glycoprotein. A peptide region (E1193 to E1269.) was found to contain HA and VN epitopes. Using both overlapping synthetic peptides and truncated fusion proteins within this region, the HA epitope defined by MAb 3D9F mapped to amino acid residues El214 to El240, while two VN epitopes defined by MAb 211391-1 and MAb 16A1 OE mapped to amino acid residues El 214 to E1233 and E1219 to E1233, respectively. The epitopes defined in this study are recognized by antibody whether or not the epitopes are glycosylated.


Mental health and the medical profession during the COVID-19 pandemic


The transmembrane metalloprotease aminopeptidase‐N (APN)/CD13 is overexpressed in various solid and hematological malignancies in humans, including acute myeloid leukemia (AML) and is thought to influence tumor progression. Here, we investigated the contribution of APN/CD13 to the regulation of growth and survival processes in AML cells in vitro. Anti‐CD13 monoclonal antibodies MY7 and SJ1D1 (which do not inhibit APN activity) and WM15 (an APN‐blocking antibody) inhibited the growth of the AML cell line U937 and induced apoptosis, as evidenced by cell accumulation in the sub‐G(1) phase, DNA fragmentation, and phosphatidylserine externalization. Isotype‐matched IgG1 and the APN/CD13 enzymatic inhibitors bestatin and 2' ,3‐dinitroflavone‐8‐acetic acid, were ineffective. Internalization of CD13‐MY7 complex into cells was followed by mitochondrial membrane depolarization, Bcl‐2 and Mcl‐1 down‐regulation, Bax up‐regulation, caspase‐9, caspase‐8, and caspase‐3 activation, and cleavage of the caspase substrate PARP‐1. The broad‐spectrum caspase inhibitor Z‐VAD‐fmk and the caspase‐9‐ and caspase‐8‐specific inhibitors significantly attenuated apoptosis. CD13 ligation also induced apoptosis and PARP‐1 cleavage in primary AML blasts, whereas normal blood cells were not affected. Overall, these data provide new evidence that CD13 can serve as a target for inducing caspase‐dependent apoptosis in AML (independently of its APN activity). These findings may have implications for tumor biology and treatment.—Piedfer, M., Dauzonne, D., Tang, R., N'Guyen, J., Billard, C., Bauvois, B. Aminopeptidase‐N/CD13 is a potential proapoptotic target in human myeloid tumor cells. FASEB J. 25, 2831‐2842 (2011). http://www.fasebj.org