title: Influenza virus morphogenesis and budding

text: Influenza viruses are enveloped, negative stranded, segmented RNA viruses belonging to Orthomyxoviridae family. Each virion consists of three major subviral components, namely (i) a viral envelope decorated with three transmembrane proteins hemagglutinin (HA), neuraminidase (NA) and M2, (ii) an intermediate layer of matrix protein (M1), and (iii) an innermost helical viral ribonucleocapsid [vRNP] core formed by nucleoprotein (NP) and negative strand viral RNA (vRNA). Since complete virus particles are not found inside the cell, the processes of assembly, morphogenesis, budding and release of progeny virus particles at the plasma membrane of the infected cells are critically important for the production of infectious virions and pathogenesis of influenza viruses as well. Morphogenesis and budding require that all virus components must be brought to the budding site which is the apical plasma membrane in polarized epithelial cells whether in vitro cultured cells or in vivo infected animals. HA and NA forming the outer spikes on the viral envelope possess apical sorting signals and use exocytic pathways and lipid rafts for cell surface transport and apical sorting. NP also has apical determinant(s) and is probably transported to the apical budding site similarly via lipid rafts and/or through cortical actin microfilaments. M1 binds the NP and the exposed RNAs of vRNPs, as well as to the cytoplasmic tails (CT) and transmembrane (TM) domains of HA, NA and M2, and is likely brought to the budding site on the piggy-back of vRNP and transmembrane proteins. Budding processes involve bud initiation, bud growth and bud release. Presence of lipid rafts and assembly of viral components at the budding site can cause asymmetry of lipid bilayers and outward membrane bending leading to bud initiation and bud growth. Bud release requires fusion of the apposing viral and cellular membranes and scission of the virus buds from the infected cellular membrane. The processes involved in bud initiation, bud growth and bud scission/release require involvement both viral and host components and can affect bud closing and virus release in both positive and negative ways. Among the viral components, M1, M2 and NA play important roles in bud release and M1, M2 and NA mutations all affect the morphology of buds and released viruses. Disassembly of host cortical actin microfilaments at the pinching-off site appears to facilitate bud fission and release. Bud scission is energy dependent and only a small fraction of virus buds present on the cell surface is released. Discontinuity of M1 layer underneath the lipid bilayer, absence of outer membrane spikes, absence of lipid rafts in the lipid bilayer, as well as possible presence of M2 and disassembly of cortical actin microfilaments at the pinching off site appear to facilitate bud fission and bud release. We provide our current understanding of these important processes leading to the production of infectious influenza virus particles.


title: A Highly Pathogenic Strain of Porcine Deltacoronavirus Caused Watery Diarrhea in Newborn Piglets

text: Porcine deltacoronavirus (PDCoV) is a newly identified virus that causes watery diarrhea in newborn piglets and results in significant economic losses to the pig industry. Since first reported in Hong Kong in 2012, PDCoV has been subsequently detected in USA, South Korea, Thailand, and mainland China. Here we isolated a strain of PDCoV, named CHN-GD-2016, from the intestinal content of a diseased newborn piglet with severe diarrhea in a pig farm in Guangdong, China. PDCoV CHN-GD-2016 could be identified by immunofluorescence with PDCoV specific rabbit antisera, and typical crown-shaped particles with spiky surface projections of this PDCoV were observed with electron microscopy. Genomic analysis showed that the PDCoV CHN-GD-2016 was closely related to other Chinese PDCoV strains, with the highest sequence similarity with the strain CHN/Tianjin/2016. Importantly, inoculation of newborn piglets with 1 × 10(5) TCID(50) of CHN-GD-2016 by oral feeding successfully reproduced clear clinical symptoms, including vomiting, dehydration, and severe diarrhea in piglets. In addition, the virus RNA in rectal swabs from 1 to 7 days post inoculation was detected, macroscopic and microscopic lesions in small intestine were observed, and viral antigen was also detected in the small intestines with immunohistochemical staining. Collectively, the data show in this study confirms that PDCoV is present in Guangdong, China and is highly pathogenic in newborn piglets.


¿Cumplimos correctamente los protocolos de prevención en oftalmología?: a propósito de la última epidemia por coronavirus


Comparative analysis of the SARS coronavirus genome: a good start to a long journey


title: Baculovirus Display: A Multifunctional Technology for Gene Delivery and Eukaryotic Library Development

text: Abstract For over a decade, phage display has proven to be of immense value, allowing selection of a large variety of genes with novel functions from diverse libraries. However, the folding and modification requirements of complex proteins place a severe constraint on the type of protein that can be successfully displayed using this strategy, a restriction that could be resolved by similarly engineering a eukaryotic virus for display purposes. The quite recently established eukaryotic molecular biology tool, the baculovirus display vector system (BDVS), allows combination of genotype with phenotype and thereby enables presentation of eukaryotic proteins on the viral envelope or capsid. Data have shown that the baculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV), is a versatile tool for eukaryotic virus display. Insertion of heterologous peptides and/or proteins into the viral surface by utilizing the major envelope glycoprotein gp64, or foreign membrane‐derived counterparts, allows incorporation of the sequence of interest onto the surface of infected cells and virus particles. A number of strategies are being investigated in order to further develop the display capabilities of AcMNPV and improve the complexity of a library that may be accommodated. Numerous expression vectors for various approaches of surface display have already been developed. Further improvement of both insertion and selection strategies toward development of a refined tool for use in the creation of useful eukaryotic libraries is, however, needed. Here, the status of baculovirus display with respect to alteration of virus tropism, antigen presentation, transgene expression in mammalian cells, and development of eukaryotic libraries will be reviewed.


title: Telehealth Availability in US Hospitals in the Face of the COVID‐19 Pandemic

text: BACKGROUND: Telehealth is likely to play a crucial role in treating COVID‐19 patients. However, not all US hospitals possess telehealth capabilities. This brief report was designed to explore US hospitals’ readiness with respect to telehealth availability. We hope to gain deeper insight into the factors affecting possession of these valuable capabilities, and how this varies between rural and urban areas. METHODS: Based on 2017 data from the American Hospital Association survey, Area Health Resource Files and Medicare cost reports, we used logistic regression models to identify predictors of telehealth and eICU capabilities in US hospitals. RESULTS: We found that larger hospitals (OR(telehealth) = 1.013; P < .01) and system members (OR(telehealth) = 1.55; P < .01) (OR(eICU) = 1.65; P < .01) had higher odds of possessing telehealth and eICU capabilities. We also found evidence suggesting that telehealth and eICU capabilities are concentrated in particular regions; the West North Central region was the most likely to possess capabilities, given that these hospitals had higher odds of possessing telehealth (OR = 1.49; P < .10) and eICU capabilities (OR = 2.15; P < .05). Rural hospitals had higher odds of possessing telehealth capabilities as compared to their urban counterparts, although this relationship was marginally significant (OR = 1.34, P < .10). CONCLUSIONS: US hospitals vary in their preparation to use telehealth to aid in the COVID‐19 battle, among other issues. Hospitals’ odds of possessing the capability to provide such services vary largely by region; overall, rural hospitals have more widespread telehealth capabilities than urban hospitals. There is still great potential to expand these capabilities further, especially in areas that have been hard hit by COVID‐19.


Permettre aux médecins et aux soignants d’exercer leur mission en accord avec leur devoir d’humanité


title: Platelet-bound antibodies detected by a flow cytometric assay in cats with thrombocytopenia

text: Summary In cats, primary or secondary immune-mediated thrombocytopenia have rarely been described or characterised. The objective of this study was to determine platelet-bound antibodies (PBA) by a flow cytometric assay in both healthy and thrombocytopenic cats. Direct PBA testing was performed in 42 thrombocytopenic cats (platelet counts 6–179×109/l, median 56×109/l). Of these 42 cats, 19 had positive PBA test results, 17 of which were considered to have secondary immune-mediated thrombocytopenia (sITP). Underlying diseases included fat necroses (four cases), feline infectious peritonitis (three), feline leukaemia virus (two) or feline immunodeficiency virus (two) infections, lymphoma (two), leukaemia (one), hepatitis (one), pyelonephritis (one), or hyperthyroidism (one). In two cats, no underlying disease was found suggesting a primary immune-mediated thrombocytopenia (pITP). The PBA test was negative in 23 cats diagnosed with varying underlying diseases and in 47 healthy control cats with platelet values within the reference range. Only seven of the 42 cats with thrombocytopenia (platelet count 10–57×109/l, median 34×109/l) had spontaneous bleeding. This study suggests that immune-mediated destruction of platelets might be an important pathological mechanism for feline thrombocytopenia caused by various underlying diseases. In cats, pITP appears to be rarely diagnosed.


title: Coronavirus (COVID‐19) in the United Kingdom: A personality‐based perspective on concerns and intention to self‐isolate

text: OBJECTIVES: Public behaviour change is necessary to contain the spread of coronavirus (COVID‐19). Based on the reinforcement sensitivity theory (RST) framework, this study presents an examination of individual differences in some relevant psychological factors. DESIGN: Cross‐sectional psychometric. METHODS: UK respondents (N = 202) completed a personality questionnaire (RST‐PQ), measures of illness attitudes, concerns about the impact of coronavirus on health services and socio‐economic infrastructures, personal safety, and likelihood of voluntary self‐isolation. RESULTS: Respondents most concerned were older, had negative illness attitudes, and scored higher on reward reactivity (RR), indicating the motivation to take positive approach action despite prevailing worry/anxiety. Personal safety concerns were highest in those with negative illness attitudes and higher fight–flight–freeze system (FFFS, reflecting fear/avoidance) scores. Results suggest people are experiencing psychological conflict: between the urge to stay safe (FFFF‐related) and the desire to maintain a normal, pleasurable (RR‐related) life. Ways of ameliorating conflict may include maladaptive behaviours (panic buying), reflecting reward‐related displacement activity. Intended self‐isolation related to FFFS, but also low behavioural inhibition system (related to anxiety) scores. Older people reported themselves less likely to self‐isolate. CONCLUSIONS: Interventions need to consider individual differences in psychological factors in behaviour change, and we discuss relevant literature to inform policy makers and communicators. STATEMENT OF CONTRIBUTION: What is already known on this subject? Reinforcement sensitivity theory (RST) personality systems can influence perception of persuasive health messages. However, there is limited evidence for their direct effects on health concerns and behaviours, and none relating to specific infectious diseases. What does this study add? Reward reactivity (RR) is associated with concern about impact of coronavirus on the NHS and other social infrastructures, indicating the motivation to take positive‐approach action despite worry/anxiety. Personal safety concerns are related to fight–flight–freeze system traits (FFFS, reflecting fear/avoidance). Intended self‐isolation related to FFFS, but also low behavioural inhibition system (related to anxiety) scores. Older people reported themselves less likely to self‐isolate. Results suggest psychological conflict: between the urge to stay safe (FFFF‐related) and the desire to maintain a normal, pleasurable life (RR‐related). Ways of ameliorating conflict may include maladaptive behaviours (panic buying), reflecting reward‐related displacement activity;


title: Sequence homology between human PARP14 and the SARS-CoV-2 ADP ribose 1’-phosphatase

text: • In support of the theory that coronaviral ADRP enzymes evolved to counteract the anti-viral immune signaling mediated by PARP14, we present sequence homology between the ADP-ribose-binding sites of the two proteins. • Interestingly this homology is even more pronounced in bat species. • We believe this model provides crucial information for the consideration of potential therapeutic interventions, based on the PARP axis in COVID-19.


title: Surgery during the COVID-19 pandemic: A comprehensive overview and perioperative care

text: Abstract COVID-19 disease is a pandemic disease caused by a single-stranded RNA virus that belongs to the coronavirus family known as 2019-nCoV (SARS-CoV-2). The disease is highly contagious and transmitted mainly by droplets or close contact. As the global incidence of COVID-19 disease is increasing dramatically, health systems worldwide started to suffer in terms of its capability to manage affected people and the ability to provide standard treatment for critically ill patients in a safe environment. As Medical staff has extensive contact with patients, families as well as other health care providers, they are very likely to cause cross-infection. This paper aims to provide a comprehensive overview of COVID-19 disease as well as to highlight essential measures that healthcare providers and surgeons need to take into consideration during their management of the patient during the COVID-19 pandemic.


COVID-19 associated Multisystem Inflammatory Syndrome in Children (MIS-C) guidelines; a Western New York approach


title: COVID‐19 and liver transplantation: Lessons learned from three reported cases

text: During the COVID-19 pandemic, transplant recipients have been recognized as more susceptible to infection, to have greater severity of disease, and prolonged shedding of this highly transmissible virus.1 However, there is limited information on the impact of COVID-19 in liver transplant (LT) recipients. We reviewed three reported cases with detailed treatment information from China to better understand the features and associated therapeutic strategies used in transplant recipients with COVID-19.2-4.


title: Essential role of HCMV deubiquitinase in promoting oncogenesis by targeting anti-viral innate immune signaling pathways

text: Cancer is a multifactorial disease and virus-mediated carcinogenesis is one of the crucial factors, which is poorly understood. Human cytomegalovirus (HCMV) is a herpesvirus and its components have been evidenced to be associated with cancer of different tissue origin. However, its role in cancer remains unknown. Here, we identified a conserved herpesviral tegument protein known as pUL48 of HCMV, encoding deubiquitinase enzyme, as having a key role in carcinogenesis. We show using deubiquitinase sufficient- and deficient-HCMV that HCMV deubiquitinase is a key in inducing enhanced cellular metabolic activity through upregulation of several anti-apoptotic genes and downregulation of several pro-apoptotic genes expression. Furthermore, HCMV deubiquitinase acquires pro-tumor functions by inhibiting PRR-mediated type I interferon via deubiquitination of TRAF6, TRAF3, IRAK1, IRF7 and STING. Taken together, our results suggest that HCMV infection may promote oncogenesis by inhibiting innate immunity of the host.


title: Amplification of Emerging Viruses in a Bat Colony

text: Bats host noteworthy viral pathogens, including coronaviruses, astroviruses, and adenoviruses. Knowledge on the ecology of reservoir-borne viruses is critical for preventive approaches against zoonotic epidemics. We studied a maternity colony of Myotis myotis bats in the attic of a private house in a suburban neighborhood in Rhineland-Palatinate, Germany, during 2008, 2009, and 2010. One coronavirus, 6 astroviruses, and 1 novel adenovirus were identified and monitored quantitatively. Strong and specific amplification of RNA viruses, but not of DNA viruses, occurred during colony formation and after parturition. The breeding success of the colony was significantly better in 2010 than in 2008, in spite of stronger amplification of coronaviruses and astroviruses in 2010, suggesting that these viruses had little pathogenic influence on bats. However, the general correlation of virus and bat population dynamics suggests that bats control infections similar to other mammals and that they may well experience epidemics of viruses under certain circumstances.


title: Bioactive natural compounds against human coronaviruses: a review and perspective

text: Coronaviruses (CoVs), a family of enveloped positive-sense RNA viruses, are characterized by club-like spikes that project from their surface, unusually large RNA genome, and unique replication capability. CoVs are known to cause various potentially lethal human respiratory infectious diseases, such as severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and the very recent coronavirus disease 2019 (COVID-19) outbreak. Unfortunately, neither drug nor vaccine has yet been approved to date to prevent and treat these diseases caused by CoVs. Therefore, effective prevention and treatment medications against human coronavirus are in urgent need. In the past decades, many natural compounds have been reported to possess multiple biological activities, including antiviral properties. In this article, we provided a comprehensive review on the natural compounds that interfere with the life cycles of SARS and MERS, and discussed their potential use for the treatment of COVID-19.


title: From SARS to MERS: crystallographic studies on coronaviral proteases enable antiviral drug design

text: This review focuses on the important contributions that macromolecular crystallography has made over the past 12 years to elucidating structures and mechanisms of the essential proteases of coronaviruses, the main protease (M(pro)) and the papain‐like protease (PL(pro)). The role of X‐ray crystallography in structure‐assisted drug discovery against these targets is discussed. Aspects dealt with in this review include the emergence of the SARS coronavirus in 2002–2003 and of the MERS coronavirus 10 years later and the origins of these viruses. The crystal structure of the free SARS coronavirus M(pro) and its dependence on pH is discussed, as are efforts to design inhibitors on the basis of these structures. The mechanism of maturation of the enzyme from the viral polyprotein is still a matter of debate. The crystal structure of the SARS coronavirus PL(pro) and its complex with ubiquitin is also discussed, as is its orthologue from MERS coronavirus. Efforts at predictive structure‐based inhibitor development for bat coronavirus M(pro)s to increase the preparedness against zoonotic transmission to man are described as well. The paper closes with a brief discussion of structure‐based discovery of antivirals in an academic setting.


title: Discovery of the First Insect Nidovirus, a Missing Evolutionary Link in the Emergence of the Largest RNA Virus Genomes

text: Nidoviruses with large genomes (26.3–31.7 kb; ‘large nidoviruses’), including Coronaviridae and Roniviridae, are the most complex positive-sense single-stranded RNA (ssRNA+) viruses. Based on genome size, they are far separated from all other ssRNA+ viruses (below 19.6 kb), including the distantly related Arteriviridae (12.7–15.7 kb; ‘small nidoviruses’). Exceptionally for ssRNA+ viruses, large nidoviruses encode a 3′-5′exoribonuclease (ExoN) that was implicated in controlling RNA replication fidelity. Its acquisition may have given rise to the ancestor of large nidoviruses, a hypothesis for which we here provide evolutionary support using comparative genomics involving the newly discovered first insect-borne nidovirus. This Nam Dinh virus (NDiV), named after a Vietnamese province, was isolated from mosquitoes and is yet to be linked to any pathology. The genome of this enveloped 60–80 nm virus is 20,192 nt and has a nidovirus-like polycistronic organization including two large, partially overlapping open reading frames (ORF) 1a and 1b followed by several smaller 3′-proximal ORFs. Peptide sequencing assigned three virion proteins to ORFs 2a, 2b, and 3, which are expressed from two 3′-coterminal subgenomic RNAs. The NDiV ORF1a/ORF1b frameshifting signal and various replicative proteins were tentatively mapped to canonical positions in the nidovirus genome. They include six nidovirus-wide conserved replicase domains, as well as the ExoN and 2′-O-methyltransferase that are specific to large nidoviruses. NDiV ORF1b also encodes a putative N7-methyltransferase, identified in a subset of large nidoviruses, but not the uridylate-specific endonuclease that – in deviation from the current paradigm - is present exclusively in the currently known vertebrate nidoviruses. Rooted phylogenetic inference by Bayesian and Maximum Likelihood methods indicates that NDiV clusters with roniviruses and that its branch diverged from large nidoviruses early after they split from small nidoviruses. Together these characteristics identify NDiV as the prototype of a new nidovirus family and a missing link in the transition from small to large nidoviruses.


title: One health – an ecological and evolutionary framework for tackling Neglected Zoonotic Diseases

text: Understanding the complex population biology and transmission ecology of multihost parasites has been declared as one of the major challenges of biomedical sciences for the 21st century and the Neglected Zoonotic Diseases (NZDs) are perhaps the most neglected of all the Neglected Tropical Diseases (NTDs). Here we consider how multihost parasite transmission and evolutionary dynamics may affect the success of human and animal disease control programmes, particularly neglected diseases of the developing world. We review the different types of zoonotic interactions that occur, both ecological and evolutionary, their potential relevance for current human control activities, and make suggestions for the development of an empirical evidence base and theoretical framework to better understand and predict the outcome of such interactions. In particular, we consider whether preventive chemotherapy, the current mainstay of NTD control, can be successful without a One Health approach. Transmission within and between animal reservoirs and humans can have important ecological and evolutionary consequences, driving the evolution and establishment of drug resistance, as well as providing selective pressures for spill‐over, host switching, hybridizations and introgressions between animal and human parasites. Our aim here is to highlight the importance of both elucidating disease ecology, including identifying key hosts and tailoring control effort accordingly, and understanding parasite evolution, such as precisely how infectious agents may respond and adapt to anthropogenic change. Both elements are essential if we are to alleviate disease risks from NZDs in humans, domestic animals and wildlife.


title: Intestinal and Peripheral Fibrinogen-Like Protein 2 Expression in Inflammatory Bowel Disease

text: BACKGROUND: Fibrinogen-like protein 2 (FGL2), a new member of the fibrinogen-like family, has recently been identified as a novel immunosuppressive molecule. AIM: The purpose of this work was to investigate intestinal and peripheral expression of FGL2 in patients with inflammatory bowel disease (IBD), mainly ulcerative colitis (UC) and Crohn’s disease (CD). METHODS: FGL2 expression in mucosal biopsies from three groups (UC group (n = 61), CD group (n = 54), and controls group (n = 35)) was detected by immunohistochemistry. Concentrations of FGL2 in plasma from 50 UC patients, 45 CD patients, and 30 controls were analyzed by enzyme-linked immunosorbent assay. Western blot of FGL2 protein and real-time fluorescent quantitative PCR of FGL2 mRNA expression by peripheral mononuclear cells was performed. Correlations of FGL2 expression with disease type, activity, and location, and with measured laboratory data, including C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), were examined. RESULTS: Intestinal and peripheral FGL2 protein data showed that FGL2 expression was significantly up-regulated in both UC and CD patients compared with controls (P < 0.001). Expression of FGL2 was higher in UC and CD patients with active disease than in those with inactive disease (P < 0.001). Moreover, FGL2 mRNA expression was significantly higher in patients with active disease than in those with inactive disease (P < 0.050). Expression of FGL2 protein was correlated with disease activity indices, CRP levels, and ESR levels. CONCLUSION: Expression of FGL2 was up-regulated in IBD patients with active disease. Measurement of FGL2 may be used as a helpful biomarker for understanding immunopathogenesis and for assessment of IBD.


title: Hepatitis C Virus Life Cycle and Lipid Metabolism

text: Hepatitis C Virus (HCV) infects over 150 million people worldwide. In most cases HCV infection becomes chronic, causing liver disease ranging from fibrosis to cirrhosis and hepatocellular carcinoma. HCV affects the cholesterol homeostasis and at the molecular level, every step of the virus life cycle is intimately connected to lipid metabolism. In this review, we present an update on the lipids and apolipoproteins that are involved in the HCV infectious cycle steps: entry, replication and assembly. Moreover, the result of the assembly process is a lipoviroparticle, which represents a peculiarity of hepatitis C virion. This review illustrates an example of an intricate virus-host interaction governed by lipid metabolism.


title: Current landscape and future prospects of antiviral drugs derived from microbial products

text: Viral infections are a major global health threat. Over the last 50 years, significant efforts have been devoted to the development of antiviral drugs and great success has been achieved for some viruses. However, other virus infections, such as epidemic influenza, still spread globally and new threats continue to arise from emerging and re-emerging viruses and drug-resistant viruses. In this review, the contributions of microbial products isolated in Institute of Microbial Chemistry for antiviral research are summarized. In addition, the current state of development of antiviral drugs that target influenza virus and hepatitis B virus, and the future prospects for antivirals from natural products are described and discussed.


High aerosol generating potential


title: Une approche intégrative pour la caractérisation initiale de l’épidémie due au virus MERS-CoV

text: Introduction L’apparition en 2012 du nouveau virus MERS-CoV au Moyen-Orient (MO) suivie de sa dissémination internationale suscite une inquiétude pour la santé publique globale. La caractérisation initiale de l’épidémie et de son potentiel de transmission a été difficile de par les incertitudes sur le réservoir et les modes de transmission. D’autre part, les données épidémiologiques étaient limitées, réduisant la puissance des analyses statistiques et de modélisation. Méthode Nous présentons ici une approche intégrative, basée sur l’analyse conjointe des cas groupés au MO et des cas importés dans la communauté européenne. Cette approche permet l’analyse la plus complète à ce stade, car elle intègre la transmission depuis un réservoir animal, la sous notification des cas et la mobilité internationale par le transport aérien. Résultats L’estimation basée sur les données collectées jusqu’au 31 août 2013 montre que l’épidémie reste sous-critique (R=0,50, 95 %IC 0,30–0,77) avec introduction d’environ 2 (95 %IC 0,8–6) nouveaux cas par transmission d’origine animale. Les infections dans la région sont pour l’heure largement dominée par cette origine zoonotique, avec une sous notification possiblement importante (ratio du nombre d’introductions estimées sur observées égal à 2,41, 95 %IC 1,03–7,32). Conclusion L’approche développée montre son intérêt pour guider la décision en santé publique, en permettant de quantifier le rôle relatif des routes de transmission, la taille vraisemblable de l’épidémie et son potentiel de dissémination international. La combinaison de différentes sources (grappes de cas, transport aérien) a permis de renforcer le potentiel explicatif de l’information disponible, et en fait un modèle utilisable pour d’autres épidémies émergentes.


title: Chiropteran types I and II interferon genes inferred from genome sequencing traces by a statistical gene-family assembler

text: BACKGROUND: The rate of emergence of human pathogens is steadily increasing; most of these novel agents originate in wildlife. Bats, remarkably, are the natural reservoirs of many of the most pathogenic viruses in humans. There are two bat genome projects currently underway, a circumstance that promises to speed the discovery host factors important in the coevolution of bats with their viruses. These genomes, however, are not yet assembled and one of them will provide only low coverage, making the inference of most genes of immunological interest error-prone. Many more wildlife genome projects are underway and intend to provide only shallow coverage. RESULTS: We have developed a statistical method for the assembly of gene families from partial genomes. The method takes full advantage of the quality scores generated by base-calling software, incorporating them into a complete probabilistic error model, to overcome the limitation inherent in the inference of gene family members from partial sequence information. We validated the method by inferring the human IFNA genes from the genome trace archives, and used it to infer 61 type-I interferon genes, and single type-II interferon genes in the bats Pteropus vampyrus and Myotis lucifugus. We confirmed our inferences by direct cloning and sequencing of IFNA, IFNB, IFND, and IFNK in P. vampyrus, and by demonstrating transcription of some of the inferred genes by known interferon-inducing stimuli. CONCLUSION: The statistical trace assembler described here provides a reliable method for extracting information from the many available and forthcoming partial or shallow genome sequencing projects, thereby facilitating the study of a wider variety of organisms with ecological and biomedical significance to humans than would otherwise be possible.


title: Measures Undertaken in China to Avoid COVID-19 Infection: Internet-Based, Cross-Sectional Survey Study

text: BACKGROUND: In early 2020, over 80,000 cases of coronavirus disease (COVID-19) were confirmed in China. Public prevention and control measures, along with efforts from all sectors of society, were undertaken to control and eliminate disease transmission. OBJECTIVE: This paper describes Chinese citizens’ response to the epidemic, the preventive measures they implemented to avoid being infected, and the public strategies that were carried out by the government, health workers, etc. We also discuss the efficacy of these measures in controlling the epidemic in China. METHODS: Information on the responses and behaviors of Chinese citizens were collected through a cross-sectional, internet-based survey using Dingxiang Doctor’s public account on WeChat. Information on public strategies implemented by all sectors of society to control the epidemic and data on new COVID-19 cases were collected from the internet, mainly from government websites. Standard descriptive statistics and multivariate logistic regression analyses were conducted to analyze the data. RESULTS: A total of 10,304 participants responded to the survey, with 10,198 valid responses; 74.1% (n=7557) were female and 25.9% (n=2641) were male. Overall, 98.2% (n=10,013) of participants paid high or very high attention to the epidemic, with WeChat being their main information source (n=9400, 92.2%). Over half the participants (n=5878, 57.7%) were confident that the epidemic could be curbed in China; 92.4% (n=9427) opened windows for ventilation more frequently than usual; 97.9% (n=9986) used masks in public; 95.7% (n=9759) avoided large crowds and stayed at home as much as possible; and 97.9% (n=9988) washed their hands more often than usual. Women were more likely to practice these behaviors than men (P<.001). With a series of strict public control measures, like nationwide health education campaigns, holiday extensions, the Examine and Approve Policy on the resumption of work, close management of working and living quarters, a health QR (Quick Response) code system, community screening, and social distancing policies, the number of new cases have decreased dramatically since February 12, 2020. CONCLUSIONS: The methods employed by Chinese citizens and authorities have effectively curtailed the spread of COVID-19, demonstrating that this pandemic can be brought under control as long as the right measures are taken.


title: Reply to “Clinical and histological characterization of vesicular COVID‐19 rashes: a prospective study in a tertiary care hospital”: Pseudoherpetic Grover disease seems to appear in patients with COVID‐19 infection

text: We have read with great interest the article by Fernandez‐Nieto et al. showing histopathological pictures of one of the two biopsied cases from their 24 patients diagnosed of vesicular lesions related to COVID‐19.(1) They additionally performed a real‐time PCR for SARS‐CoV‐2 that was negative in four tested cases.(1) Regarding their case histopathological pictures, we agree with their description as we can observe acantholytic cells, some of them with dyskeratotic features or grouped without a clear molding of nucleus but reminiscent of the multinucleated giant cells typically observed in herpetic lesions, a picture that looks to our eyes, most likely as an infrequent entity named pseudoherpetic Grover disease (GD),(2) first described as vesicular GD by Fernandez‐Figueras.(3)


title: Molecular conservation and differential mutation on ORF3a gene in Indian SARS-CoV2 genomes

text: Abstract A global emergency due to the COVID-19 pandemic demands various studies related to genes and genomes of the SARS-CoV2. Among other important proteins, the role of accessory proteins are of immense importance in replication, regulation of infections of the coronavirus in the hosts. The largest accessory protein in the SARS-CoV2 genome is ORF3a which modulates the host response to the virus infection and consequently it plays an important role in pathogenesis. In this study, an attempt is made to decipher the conservation of nucleotides, dimers, codons and amino acids in the ORF3a genes across thirty-two genomes of Indian patients. ORF3a gene possesses single and double point mutations in Indian SARS-CoV2 genomes suggesting the change of SARS-CoV2's virulence property in Indian patients. We find that the parental origin of the ORF3a gene over the genomes of SARS-CoV2 and Pangolin-CoV is same from the phylogenetic analysis based on conservation of nucleotides and so on. This study highlights the accumulation of mutation on ORF3a in Indian SARS-CoV2 genomes which may provide the designing therapeutic approach against SARS-CoV2.


title: Quantitative prediction of mouse class I MHC peptide binding affinity using support vector machine regression (SVR) models

text: BACKGROUND: The binding between peptide epitopes and major histocompatibility complex proteins (MHCs) is an important event in the cellular immune response. Accurate prediction of the binding between short peptides and the MHC molecules has long been a principal challenge for immunoinformatics. Recently, the modeling of MHC-peptide binding has come to emphasize quantitative predictions: instead of categorizing peptides as "binders" or "non-binders" or as "strong binders" and "weak binders", recent methods seek to make predictions about precise binding affinities. RESULTS: We developed a quantitative support vector machine regression (SVR) approach, called SVRMHC, to model peptide-MHC binding affinities. As a non-linear method, SVRMHC was able to generate models that out-performed existing linear models, such as the "additive method". By adopting a new "11-factor encoding" scheme, SVRMHC takes into account similarities in the physicochemical properties of the amino acids constituting the input peptides. When applied to MHC-peptide binding data for three mouse class I MHC alleles, the SVRMHC models produced more accurate predictions than those produced previously. Furthermore, comparisons based on Receiver Operating Characteristic (ROC) analysis indicated that SVRMHC was able to out-perform several prominent methods in identifying strongly binding peptides. CONCLUSION: As a method with demonstrated performance in the quantitative modeling of MHC-peptide binding and in identifying strong binders, SVRMHC is a promising immunoinformatics tool with not inconsiderable future potential.


title: Detection of transmissible gastroenteritis virus using cDNA probes

text: Five cDNA probes prepared from molecular clones representing genomic RNA sequences of the virulent Miller strain of transmissible gastroenteritis virus (TGEV) were used in a dot blot hybridization assay to detect TGEV in cell culture and fecal specimens. Two clones (pA2 and pB4) represent nucleotide base pairs at the 3′ terminus of the Miller TGEV genome. The other three clones represent various portions of the 5′ end of the E 2 gene, which codes for the major surface glycoprotein of TGEV. Each of the(32)P-labeled cDNA probes hybridized to the virulent Miller, attenuated Purdue and four field strains of TGEV. The probes detected 200 to 2000 pg of TGEV RNA extracted from density gradient purified virions and did not hybridize RNA from mock-infected cell cultures, porcine rotavirus or antigenically unrelated coronaviruses. The pB4 and Hpa-1600 probes detected TGEV RNA sequences in 79 and 88%, respectively of 34 field samples identified as TGEV positive by the immunofluorescence assay and electron microscopy (EM). The pD24 clone, which is able to differentiate TGEV from the antigenically related coronaviruses, also compared favorably with conventional methods of EM and immunofluorescence for the detection of TGEV in fecal specimens.


title: A Likelihood-based Index of Protein–Protein Binding Affinities with Application to Influenza HA Escape from Antibodies

text: In many biological systems, proteins interact with other organic molecules to produce indispensable functions, in which molecular recognition phenomena are essential. Proteins have kept or gained their functions during molecular evolution. Their functions seem to be flexible, and a few amino acid substitutions sometimes cause drastic changes in function. In order to monitor and predict such drastic changes in the early stages in target populations, we need to identify patterns of structural changes during molecular evolution causing decreases or increases in the binding affinity of protein complexes. In previous work, we developed a likelihood-based index to quantify the degree to which a sequence fits a given structure. This index was named the sequence-structure fitness (SSF) and is calculated empirically based on amino acid preferences and pairwise interactions in the structural environment present in template structures. In the present work, we used the SSF to develop an index to measure the binding affinity of protein–protein complexes defined as the log likelihood ratio, contrasting the fitness of the sequences to the structure of the complex and that of the uncomplexed proteins. We applied the developed index to the complexes formed between influenza A hemagglutinin (HA) and four antibodies. The antibody–antigen binding region of HA is under strong selection pressure by the host immune system. Hence, examination of the long-term adaptation of HA to the four antibodies could reveal the strategy of the molecular evolution of HA. Two antibodies cover the HA receptor-binding region, while the other two bind away from the receptor-binding region. By focusing on branches with a significant decline in binding ability, we could detect key amino acid replacements and investigate the mechanism via conditional probabilities. The contrast between the adaptations to the two types of antibodies suggests that the virus adapts to the immune system at the cost of structural change.


title: Replacement of fish meal in cobia (Rachycentron canadum) diets using an organically certified protein

text: Abstract A six-week feeding trial was conducted to evaluate the use of a yeast-based, certified organic protein source as a replacement for fish meal in diets for cobia. Five experimental diets were formulated to provide 40% crude protein and 11% dietary lipid (dry matter basis) with the yeast-based protein source replacing Special Select® menhaden fish meal at 25%, 50%, 75% and 100% of dietary protein. Ten juvenile cobia (initial weight 11.5 g/fish) were randomly stocked in triplicate 300 l circular fiberglass tanks (n =30 treatment−1) and hand-fed the diets based upon total tank biomass two times daily at 0900 and 1400 h. Fish were group weighed weekly to monitor performance and adjust feeding rations. Water temperature and salinity were maintained at 27 °C and 15‰, respectively. At the end of the feeding trial, weight gain, ranging from 86% to 512%, and feed conversion ratio values, ranging from 1.9 to 5.8, were significantly affected by the inclusion of the yeast-based protein source, with decreasing values as inclusion levels of the yeast-based protein source rose above 25% of dietary protein. Cobia fed the diet containing 25% of dietary protein from the yeast-based protein source had equal weight gain and feed conversion ratio values as fish fed the control diet composed of 100% fish meal (503 vs. 512 and 1.9 vs. 1.9, respectively). Biological indices including hepatosomatic index, visceral somatic index and muscle ratio were all similarly affected by inclusion of the yeast-based protein source, with significant impacts when inclusion levels rose above 25% of dietary protein. As with the weight gain and feed efficiency ratio values, fish fed the diet containing 25% of protein from the yeast-based source had similar values as those observed in the control animals. This study represents the first attempt to utilize an organically certified protein source as a replacement for fish meal in diets for juvenile cobia. Although levels of inclusion of the yeast-based protein source above 50% of dietary protein resulted in detrimental effects on production characteristics, the data clearly suggest that, at a minimum, 25% of dietary protein can be provided by this yeast-based protein in diets for cobia.


title: Decreased interleukin-18 response in asthmatic children with severe Mycoplasma pneumoniae pneumonia

text: Abstract Purpose Mycoplasma pneumoniae (M. pneumoniae) is a common causative agent of pneumonia in children. The aim of this study is to determine whether there is any difference in selected cytokine or chemokines response in asthmatic children compared to non-asthmatic children during acute M. pneumoniae pneumonia. Methods Seventy-five children, 6–12years of age, admitted with M. pneumoniae pneumonia were enrolled. Two patient groups were defined: the children with known asthma (N =40) and non-asthmatic children (N =35). Interleukin (IL)-18 and selected chemokines, IL-8, CXCL9, CXCL10, and regulation upon activation normal T-cell expressed and secreted (RANTES) were measured by means of ELISA in the plasma samples of the patients collected on admission. We investigated the values of these mediators in relation to the asthma status and symptom severity of the patients. Twenty age-matched, non-infected controls were also studied. Results Plasma levels of IL-18 and the chemokines increased significantly in the patients with M. pneumoniae pneumonia compared to non-infected, age-matched controls (P <0.01). However, the asthmatic patients showed significantly reduced IL-18 and CXCL10 responses (P <0.01, <0.05, respectively) and had more severe pneumonia symptoms (P <0.01) compared to non-asthmatic patients. IL-18 was significantly lower in severe pneumonia group than in non-severe group (P <0.05). Conclusions Our study suggests that IL-18 and the chemokines are importantly involved in the pathogenesis of M. pneumoniae pneumonia. It also indicates that some asthmatic children have deficient IL-18 response when affected by M. pneumoniae pneumonia, which might be associated with more severe pneumonia observed in this group of patients.


title: Strengthening global health security by embedding the International Health Regulations requirements into national health systems

text: The International Health Regulations (IHR) 2005, as the overarching instrument for global health security, are designed to prevent and cope with major international public health threats. But poor implementation in countries hampers their effectiveness. In the wake of a number of major international health crises, such as the 2014 Ebola and 2016 Zika outbreaks, and the findings of a number of high-level assessments of the global response to these crises, it has become clear that there is a need for more joined-up thinking between health system strengthening activities and health security efforts for prevention, alert and response. WHO is working directly with its Member States to promote this approach, more specifically around how to better embed the IHR (2005) core capacities into the main health system functions. This paper looks at how and where the intersections between the IHR and the health system can be best leveraged towards developing greater health system resilience. This merging of approaches is a key component in pursuit of Universal Health Coverage and strengthened global health security as two mutually reinforcing agendas.


title: The RNA of human coronavirus OC-43

text: Abstract A homogeneous RNA complex with a sedimentation coefficient of 70 S and an apparent molecular weight of approximately 6.1 × 106 was released from purified 32P-labeled, mouse-brain-derived OC-43 virus after treatment with 1% sodium dodecyl sulfate (SDS) for 15 min at 23°. The complex was highly susceptible to heat, releasing 4 S RNA fragments at 37° and breaking down to fragments of 4–70 S at 60°; it was also degraded by centrifugation through dimethyl sulfoxide gradients. Unlike tobacco mosaic virus or Rous sarcoma virus RNA, OC-43 RNA prepared by extraction with phenol-SDS or phenol-chloroform degraded into a range of fragments with coefficients of 15–55 S; 4 S RNA was also present as a minor component. This suggests that (a) extensive nicking of a large RNA molecule has occurred during viral growth, due to ribonucleases which are inactivated during phenol extractions; (b) heterogeneity for OC-43 RNA is not due to internal ribonuclease activity and fragments are held together by noncovalent linkages much weaker than those present in the 70 S retroviral RNA complex, or by small proteins; or, most probably, (c) a combination of extensive nicking and weak noncovalent linkages results in the heterogeneous denaturation products.


title: Diagnostic accuracy of the Rivalta test for feline infectious peritonitis

text: BACKGROUND: The Rivalta test has been used routinely in Europe to diagnose feline infectious peritonitis (FIP) in cats with effusions, but its diagnostic accuracy is uncertain. OBJECTIVES: The objectives of this study were to calculate sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values of the Rivalta test for FIP and to identify correlations between a positive Rivalta test and variables measured in effusion fluid and peripheral blood. METHODS: In this retrospective study, medical records of cats with effusions were reviewed, and cats with conclusive results for the Rivalta test were included. The prevalence of FIP in this population was determined, and sensitivity, specificity, and PPV and NPV of the Rivalta test were calculated. Variables measured in effusion fluid and peripheral blood were compared between cats that had positive or negative Rivalta tests using the Mann–Whitney U‐test and multivariate analysis. RESULTS: Of 851 cats with effusions, 782 had conclusively positive or negative results for the Rivalta test. A definitive final diagnosis was made in 497 of these cats. Prevalence of FIP in cats with effusion and a conclusive Rivalta test result was 34.6%. The Rivalta test had a sensitivity of 91.3%, specificity of 65.5%, PPV of 58.4%, and NPV of 93.4% for the diagnosis of FIP. These values increased when cats with lymphoma or bacterial infections were excluded, or when only cats ≤ 2 years were considered. Increased effusion cholesterol concentration and specific gravity as well as decreased serum albumin:globulin ratio and hyperbilirubinemia were positively correlated with positive Rivalta test results. CONCLUSIONS: Sensitivity, specificity, and PPV of the Rivalta test for the diagnosis of FIP were lower than previously reported except when used in young cats. The components in effusions that lead to a positive Rivalta test remain unknown, but the positivity is not simply related to high total protein concentration.


title: Effects of COVID-19 Infection during Pregnancy and Neonatal Prognosis: What Is the Evidence?

text: Background: This study’s aims are to assess the current evidence presented in the literature regarding the potential risks of COVID-19 infection among pregnant women and consequent fetal transmission. Methods: a systematic literature review assessing papers published in the most comprehensive databases in the field of health intended to answer the question, “What are the effects of COVID-19 infection during pregnancy, and what is the neonatal prognosis?” Results: 49 papers published in 2020 were eligible, presenting low levels of evidence. A total of 755 pregnant women and 598 infants were assessed; more than half of pregnant women had C-sections (379/65%). Only 493 (82%) infants were tested for SARS-CoV-2, nine (2%) of whom tested positive. There is, however, no evidence of vertical transmission based on what has been assessed so far, considering there are knowledge gaps concerning the care provided during and after delivery, as well as a lack of suitable biological samples for testing SARS-CoV-2. Conclusions: We cannot rule out potential worsening of the clinical conditions of pregnant women infected with SARS-CoV-2, whether the infection is associated with comorbidities or not, due to the occurrence of respiratory disorders, cardiac rhythm disturbances, and acid-base imbalance, among others. We recommend relentless monitoring of all pregnant women in addition to testing them before delivery or the first contact with newborns.


title: Ethyl alcohol threshold test: a fast, reliable and affordable olfactory Assessment tool for COVID-19 patients

text: OBJECTIVE: COVID-19 patients may present mild symptoms. The identification of paucisymptomatic patients is paramount in order to interrupt the transmission chain of the virus. Olfactory loss could be one of those early symptoms which might help in the diagnosis of COVID-19 patients. In this study, we aim to develop and validate a fast, inexpensive, reliable and easy-to-perform olfactory test for the screening of suspected COVID-19 patients. STUDY DESIGN: Phase I was a case–control study and Phase II a transversal descriptive study. SUBJECTS AND METHODS: Olfaction was assessed with the ethyl alcohol threshold test and symptoms with visual analogue scales. The study was designed in two phases: In Phase I, we compared confirmed COVID-19 patients and healthy controls. In Phase II, patients with suspected COVID-19 infection referred for testing were studied. RESULTS: 275 participants were included in Phase I, 135 in Phase II. The ROC curve showed an AUC of 0.749 in Phase I, 0.737 in Phase II. The cutoff value which offered the highest amount of correctly classified patients was ≥ 2 (10% alcohol) for all age intervals. The odds ratio was 8.19 in Phase I, 6.56 in Phase II with a 75% sensitivity. When cases report normal sense of smell (VAS < 4), it misdiagnoses 57.89% of patients detected by the alcohol threshold test. CONCLUSION: The olfactory loss assessed with the alcohol threshold test has shown high sensitivity and odds ratio in both patients with confirmed COVID-19 illness and participants with suspected SARS-CoV-2 infection.


title: Epidemiological study of hemotropic mycoplasmas (hemoplasmas) in cats from central Spain

text: BACKGROUND: Hemotropic mycoplasmas (hemoplasmas) have been found infecting cats worldwide. However, studies about feline hemoplasma infections in Spain are scarce. Therefore, the purpose of the research was to evaluate the prevalence of feline hemotropic mycoplasmas and to characterize risk factors and clinical findings associated with these infections in a cat population from the Madrid area, Spain. METHODS: Polymerase chain reaction (PCR) was employed to detect Mycoplasma haemofelis (Mhf), “Candidatus Mycoplasma haemominutum” (CMhm) and “Candidatus Mycoplasma turicensis” (CMt) in blood samples from 456 client-owned and 138 stray cats from Madrid. In order to assess associations between these hemoplasma infections and epidemiological parameters, data regarding signalment, environment, prophylaxis measures, retrovirus status, clinical signs and laboratory findings were compiled, whenever possible. RESULTS: DNA of feline hemoplasmas was detected from the blood of 63 out of 594 cats (10.6%), with a prevalence of 3.7% (22/594) for Mhf, 8.1% (48/594) for CMhm and 0.5% (3/594) for CMt. Stray cats had statistically higher prevalences of feline hemoplasmas (15.9%) and, specifically, of Mhf (8.7%) than client-owned cats (9 and 2.2%, respectively). A total of seven cats (1.17%) were co-infected with “Candidatus M. haemominutum” and M. haemofelis, two (0.33%) with “Candidatus M. haemominutum” and “Candidatus M. turicensis” and another one (0.17%) with M. haemofelis and Candidatus “M. turicensis”. Male gender, collection of blood during warm months and FeLV/FIV positivity status were associated with hemotropic mycoplasma infection in cats from Madrid. Additionally, within the group of client-owned cats, hemoplasma infection was associated with adult age, outdoor access, and the existence of low haematocrit, erythrocyte count and haemoglobin concentration values. CONCLUSIONS: To our knowledge, this is the first epidemiological survey of feline hemoplasmas performed in central Spain (Madrid). Our study confirms that “Ca. Mycoplasma haemominutum”, Mycoplasma haemofelis and “Ca. Mycoplasma turicensis” are infecting client-owned and stray cats in this region of Spain, “Ca. Mycoplasma haemominutum” being the most prevalent species. More studies are necessary to help understand the role of the natural infection by these species of hemoplasma in cats.


title: Graft-vs-tumor effect in patients with advanced nasopharyngeal cancer treated with nonmyeloablative allogeneic PBSC transplantation

text: While nonmyeloablative peripheral blood stem cell transplantation (NST) has shown efficacy against several solid tumors, it is untested in nasopharyngeal cancer (NPC). In a phase II clinical trial, 21 patients with pretreated metastatic NPC underwent NST with sibling PBSC allografts, using CY conditioning, thymic irradiation and in vivo T-cell depletion with thymoglobulin. Stable lymphohematopoietic chimerism was achieved in most patients and prophylactic CYA was tapered at a median of day +30. Seven patients (33%) showed partial response and three (14%) achieved stable disease. Four patients were alive at 2 years and three showed prolonged disease control of 344, 525 and 550 days. With a median follow-up of 209 (4–1147) days, the median PFS was 100 days (95% confidence interval (CI), 66–128 days), and median OS was 209 days (95% CI, 128–236 days). Patients with chronic GVHD had better survival—median OS 426 days (95% CI, 194–NE days) vs 143 days (95% CI, 114–226 days) (P=0.010). Thus, NST may induce meaningful clinical responses in patients with advanced NPC.


title: Genetic susceptibility to infectious diseases: big is beautiful, but will bigger be even better?

text: Genetic epidemiology, including twin studies, provides robust evidence that genetic variation in human populations contributes to susceptibility to infectious disease. One of the major limitations of studies that attempt to identify the genes and mechanisms that underlie this susceptibility has been lack of power caused by small sample size. With the development of novel technologies, burgeoning information on the human genome, the HapMap project, and human genetic diversity, we are at the beginning of a new era in the study of the genetics of complex diseases. This review looks afresh at the epidemiological evidence that supports a role for genetics in susceptibility to infectious disease, examines the somewhat limited achievements to date, and discusses current advances in methodology and technology that will potentially lead to translational data in the future.


title: Infectious Bronchitis Virus Regulates Cellular Stress Granule Signaling

text: Viruses must hijack cellular translation machinery to express viral genes. In many cases, this is impeded by cellular stress responses. These stress responses result in the global inhibition of translation and the storage of stalled mRNAs, into RNA-protein aggregates called stress granules. This results in the translational silencing of the majority of mRNAs excluding those beneficial for the cell to resolve the specific stress. For example, the expression of antiviral factors is maintained during viral infection. Here we investigated stress granule regulation by Gammacoronavirus infectious bronchitis virus (IBV), which causes the economically important poultry disease, infectious bronchitis. Interestingly, we found that IBV is able to inhibit multiple cellular stress granule signaling pathways, whilst at the same time, IBV replication also results in the induction of seemingly canonical stress granules in a proportion of infected cells. Moreover, IBV infection uncouples translational repression and stress granule formation and both processes are independent of eIF2α phosphorylation. These results provide novel insights into how IBV modulates cellular translation and antiviral stress signaling.


title: The Prevalence and Significance of HTLV-I/II Seroindeterminate Western Blot Patterns

text: Human T-lymphotropic virus type I (HTLV-I) infects an estimated 15–20 million persons worldwide. A number of diseases have been associated with the virus including adult T-cell leukemia (ATL), HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP), HTLV-I uveitis, and HTLV-I-associated infective dermatitis. Once it was shown that there is an increased risk for developing HAM/TSP associated with blood transfusion, screening for HTLV-1 among blood banks was implemented in Japan, United States, France, and the Netherlands. This process includes detection by an enzyme immunoassay (EIA) followed by a confirmatory Western blot (WB) in which recombinant proteins specific for HTLV-I Env glycoproteins are incorporated into WB strips. HTLV-I seropositive results are defined by the presence of antibodies against either gp46 or gp62/68 (both Env protein bands) and either p19, p24, or p53 (one of the gag bands). HTLV-II seropositivity is confirmed by the presence of rgp46-II. However, numerous cases have been documented in which serum samples are reactive by EIA, but an incomplete banding pattern is displayed by subsequent confirmatory WB. Although the significance of these HTLV-I/II seroindeterminates is unclear, it may suggest a much higher incidence of exposure to HTLV-I/II than previously estimated.


title: The Nucleoprotein and Phosphoprotein of Peste des Petits Ruminants Virus Inhibit Interferons Signaling by Blocking the JAK-STAT Pathway

text: Peste des petits ruminants virus (PPRV) is associated with global peste des petits ruminants resulting in severe economic loss. Peste des petits ruminants virus dampens host interferon-based signaling pathways through multiple mechanisms. Previous studies deciphered the role of V and C in abrogating IFN-β production. Moreover, V protein directly interacted with signal transducers and activators of transcription 1 (STAT1) and STAT2 resulting in the impairment of host IFN responses. In our present study, PPRV infection inhibited both IFN-β- and IFN-γ-induced activation of IFN-stimulated response element (ISRE) and IFN-γ-activated site (GAS) element, respectively. Both N and P proteins, functioning as novel IFN response antagonists, markedly suppressed IFN-β-induced ISRE and IFN-γ-induced GAS promoter activation to impair downstream upregulation of various interferon-stimulated genes (ISGs) and prevent STAT1 nuclear translocation. Specifically, P protein interacted with STAT1 and subsequently inhibited STAT1 phosphorylation, whereas N protein neither interacted with STAT1 nor inhibited STAT1 phosphorylation as well as dimerization, suggesting that the N and P protein antagonistic effects were different. Though they differed in their relationship to STAT1, both proteins blocked JAK-STAT signaling, severely negating the host antiviral immune response. Our study revealed a new mechanism employed by PPRV to evade host innate immune response, providing a platform to study the interaction of paramyxoviruses and host response.


title: SARS-CoV-2-Pandemie und ihre Auswirkungen auf Orthopädie und Unfallchirurgie: „Booster“ für die Telemedizin

text: With the outbreak of the COVID-19 pandemic, interest in telemedical care options has grown. Today’s telemedicine options relate not only to the area of diagnostics, but also to the monitoring of therapy courses and rehabilitation measures. Due to the current resource constraints, many orthopedic surgeons and trauma surgeons have been forced to set up video consultations, although standards for orthopedic video consultations have so far been insufficiently developed. In order to maximize the efficiency of virtual orthopedics, patients should be prepared for the virtual visit (checklist with specific instructions on camera positioning, body positioning, adjustment and clothing, testing of audiovisual skills). Classic diagnostic measures such as anamnesis, inspection and assessment of radiological findings are possible in the video consultation. One major disadvantage is, that there is no possibility for a functional examination (stability of the knee joint, shoulder tests). Although initial controlled scientific studies have shown that telemedical diagnostics are not inferior to conventional diagnostics, there are still no validated examination protocols and methods. Postoperative monitoring of rehabilitation measures can, for example, be made easier by the use of sensors. With modern sensors, inexpensive detection of joint mobility and joint position is now possible and is already used in the area of rehabilitation after reconstruction of the anterior cruciate ligament. Here too, further research is certainly necessary to validate these methods. We believe that the current pandemic offers opportunities to further expand the possibilities of telemedicine for orthopedics and trauma surgery in order to continue to use it in the future (e.g. in the care of patients from abroad or in sparsely populated areas, care of professional athletes).


title: Interactive influence of infectious disease and genetic diversity in natural populations

text: Abstract The importance of infectious disease in the survival and adaptation of animal populations is rapidly becoming apparent. Throughout evolution, animal species have been continually afflicted with devastating disease outbreaks which have influenced the demographic and genetic status of the populations. Some general population consequences of such epidemics include selection for disease resistance, the occasional alteration of host gene frequencies by a genetic ‘founder effect’ after an outbreak, and genetic adaptation of parasites to abrogate host defense mechanisms. A wide variety of host cellular genes which are polymorphic within species and which confer a regulatory effect on the outcome of infectious diseases has recently been discovered. The critical importance of maintaining genetic diversity with respect to disease defense genes in natural populations is indicated by certain populations which have reduced genetic variability and apparent increased vulnerability to infectious disease.


title: The heptad repeat region is a major selection target in MERS-CoV and related coronaviruses

text: Middle East respiratory syndrome coronavirus (MERS-CoV) originated in bats and spread to humans via zoonotic transmission from camels. We analyzed the evolution of the spike (S) gene in betacoronaviruses (betaCoVs) isolated from different mammals, in bat coronavirus populations, as well as in MERS-CoV strains from the current outbreak. Results indicated several positively selected sites located in the region comprising the two heptad repeats (HR1 and HR2) and their linker. Two sites (R652 and V1060) were positively selected in the betaCoVs phylogeny and correspond to mutations associated with expanded host range in other coronaviruses. During the most recent evolution of MERS-CoV, adaptive mutations in the HR1 (Q/R/H1020) arose in camels or in a previous host and spread to humans. We determined that different residues at position 1020 establish distinct inter- and intra-helical interactions and affect the stability of the six-helix bundle formed by the HRs. A similar effect on stability was observed for a nearby mutation (T1015N) that increases MERS-CoV infection efficiency in vitro. Data herein indicate that the heptad repeat region was a major target of adaptive evolution in MERS-CoV-related viruses; these results are relevant for the design of fusion inhibitor peptides with antiviral function.


title: Ageing and life-long maintenance of T-cell subsets in the face of latent persistent infections

text: A diverse and well-balanced repertoire of T cells is believed to be crucial for the efficacious defence against infection with new or re-emerging pathogens throughout life. In the last third of the mammalian lifespan, the maintenance of a balanced T-cell repertoire becomes highly challenging due to changes in T-cell production and consumption. In this Review, I ask whether latent persistent pathogens might be key factors that drive this imbalance and whether they determine the extent of age-associated immune deficiency


title: DOVE: An Infectious Disease Outbreak Statistics Visualization System

text: Humans are susceptible to various infectious diseases. However, humanity still has limited responses to emergent and recurrent infectious diseases. Recent developments in medical technology have led to various vaccines being developed, but these vaccines typically require a considerable amount of time to counter infectious diseases. Therefore, one of the best methods to prevent infectious diseases is to continuously update our knowledge with useful information from infectious disease information systems and taking active steps to safeguard ourselves against infectious diseases. Some existing infectious disease information systems simply present infectious disease information in the form of text or transmit it via e-mail. Other systems provide data in the form of files or maps. Most existing systems display text-centric information regarding infectious disease outbreaks. Therefore, understanding infectious disease outbreak information at a glance is difficult for users. In this paper, we propose the infectious disease outbreak statistics visualization system, called to DOVE, which collects infectious disease outbreak statistics from the Korea Centers for Disease Control & Prevention and provides statistical charts with district, time, infectious disease, gender, and age data. Users can easily identify infectious disease outbreak statistics at a glance by simply entering the district, time, and name of an infectious disease into our system. Additionally, each statistical chart allows users to recognize the characteristics of an infectious disease and predict outbreaks by investigating the outbreak trends of that disease. We believe that our system provides effective information to help prevent infectious disease outbreaks. Our system is currently available on the web at http://www.epidemic.co.kr/statistics.


title: Transfusions for anemia in adult and pediatric patients with malignancies

text: Abstract Anemia is present in over two-thirds of patients with malignant hematological disorders. The etiology of anemia predominates from ineffective erythropoiesis from marrow infiltration, cytokine related suppression, erythropoietin suppression, and vitamin deficiency; ineffective erythropoiesis is further exacerbated by accelerated clearance due to antibody mediated hemolysis and thrombotic microangiopathy. As the anemia is chronic in nature, symptoms are generally well tolerated and often non-specific. Transfusion of red blood cells (RBCs) is a balance between providing benefit for patients while avoiding risks of transfusion. Conservative/restrictive RBC transfusion practices have shown equivalent patient outcomes compared to liberal transfusion practices, and meta-analysis has shown improved in-hospital mortality, reduced cardiac events, re-bleeding, and bacterial infections. The implications for a lower threshold for transfusion in patients with malignancies are therefore increasingly being scrutinized. Alternative management strategies for anemia with IV iron and erythropoietin stimulating agents (ESAs) should be considered in the appropriate settings.


title: H1N1 Influenza Pandemic in Italy Revisited: Has the Willingness to Get Vaccinated Suffered in the Long Run?

text: BACKGROUND: The aim of the study is to assess the long-term secondary effects of personal experience with the H1N1 pandemic of 2009/2010 and the perception of the institutional reaction to it on Italians’ willingness to get vaccinated in case of a novel influenza pandemic. DESIGN AND METHODS: We conducted 140 face-to-face interviews in the Registry Office of the Municipality of Milan, Italy, from October to December 2012. RESULTS: Willingness to get vaccinated during a novel influenza pandemic was best predicted by having been vaccinated against the seasonal flu in the past (OR=5.18; 95%CI: 1.40 to 19.13) and fear of losing one’s life in case of an infection with H1N1 (OR=4.09; 95%CI: 1.68 to 9.97). It was unaffected by the assessment of institutional performance. CONCLUSIONS: The findings of this study do not point to long-term secondary effects of the institutional handling of the H1N1 pandemic. The results highlight the fact that behavioural intention is not the same as behaviour, and that the former cannot simply be taken as an indicator of the latter.


title: Highlights of traditional Chinese medicine frontline expert advice in the China national guideline for COVID-19

text: Abstract Introduction The World Health Organization has declared the coronavirus disease (COVID-19) as a pandemic on 11 March 2020, after the number of confirmed cases outside China increased 13-fold. As the epicentre of the initial outbreak, China has been updating the National COVID-19 Diagnostic and Treatment Guideline with up-to-date information about the disease. To facilitate the implementation of integrative Chinese–Western Medicine in COVID-19 management, Traditional Chinese medicine (TCM) has been recommended in recent editions of the national guideline. Methods The national guideline summarised the opinions and frontline experience of medical experts across the country to provide by far the best management for COVID-19. We extracted the case definition and clinical classifications of COVID-19 in China along with relevant TCM treatments cited in the seventh edition of the guideline, with an intend to disseminate practical information to TCM practitioners and researchers around the world. Results We presented the most recent case definition, clinical classifications, and relevant TCM treatments of COVID-19 in accordance to recommendations of the Chinese guideline. TCM treatments are stratified into two groups based on patients’ disease status. Four types of Chinese patent medicines are recommended for suspected COVID-19 cases. Several herbal formulae are recommended for confirmed COVID-19 cases according to their clinical classification and TCM pattern diagnoses. Two herbal formulae are also recommended for rehabilitation of recovering cases. Conclusion To control the waves of COVID-19 outbreak, countries must ensure the adherence of their citizens to local public health measures. Medical professionals should diagnose and treat patients according to up-to-date guidelines. Future evaluation of the outcomes of implementing TCM recommendations will strengthen the evidence base for COVID-19 management for the sake of public health and the internationalisation of TCM.


title: Critically ill SARS-CoV-2 patients display lupus-like hallmarks of extrafollicular B cell activation

text: Wide heterogeneity of disease course ranging from asymptomatic spread to respiratory failure and death has become a hallmark of the SARS-CoV-2 pandemic. While this clinical spectrum is well documented, its immunologic underpinnings are less clear. We have therefore, initiated studies of the B cell responses as they would participate in both early effector responses and in the initiation of memory formation. In terms of effector responses, we were particularly interested in the engagement and clinical correlates of the extra-follicular pathway (EF), we recently described in flaring SLE. In this systemic autoimmune disease, the EF pathway is initiated by newly activated naive B cell (aN) leading to large expansion of autoantibody-producing antibody-secreting cells through the generation of an epigenetically primed B cell precursor which are double negative (DN) for naive (IgD) and memory markers (CD27) and lacking expression of CXCR5 and CD21 (DN2). These highly activated D2 cells are also distinguished by high expression of CD11c and T-bet and are TLR7-driven. Both, TLR7-stimulation which is triggered by ssRNA and the central role played by their murine counterparts (typically characterized as Age-Associated B cells), in viral clearance, strongly supported the hypothesis that DN2 cells and the global EF pathway could be prominently engaged in COVID-19 patients. Also of note, EF B cell activation is particularly prominent in SLE patients of African-American ancestry, a population disproportionately represented in severe COVID-19. In this study we find that critically-ill patients with COVID-19 robustly upregulate constituents of the extrafollicular pathway, produce enormous numbers of antibody secreting cells, and lose unique transitional B cell populations that correlate with positive prognosis. This patient cluster associates tightly with biomarkers of poor outcomes and exhibits high rates of mortality. Thus, this B cell phenotype might serve as an immunological marker of severe COVID infection at early stages and could therefore identify a patient subset likely to benefit from targeted immunomodulatory therapy aimed at alleviating disease burden.


title: Analysis of Cathepsin and Furin Proteolytic Enzymes Involved in Viral Fusion Protein Activation in Cells of the Bat Reservoir Host

text: Bats of different species play a major role in the emergence and transmission of highly pathogenic viruses including Ebola virus, SARS-like coronavirus and the henipaviruses. These viruses require proteolytic activation of surface envelope glycoproteins needed for entry, and cellular cathepsins have been shown to be involved in proteolysis of glycoproteins from these distinct virus families. Very little is currently known about the available proteases in bats. To determine whether the utilization of cathepsins by bat-borne viruses is related to the nature of proteases in their natural hosts, we examined proteolytic processing of several viral fusion proteins in cells derived from two fruit bat species, Pteropus alecto and Rousettus aegyptiacus. Our work shows that fruit bat cells have homologs of cathepsin and furin proteases capable of cleaving and activating both the cathepsin-dependent Hendra virus F and the furin-dependent parainfluenza virus 5 F proteins. Sequence analysis comparing Pteropus alecto furin and cathepsin L to proteases from other mammalian species showed a high degree of conservation; however significant amino acid variation occurs at the C-terminus of Pteropus alecto furin. Further analysis of furin-like proteases from fruit bats revealed that these proteases are catalytically active and resemble other mammalian furins in their response to a potent furin inhibitor. However, kinetic analysis suggests that differences may exist in the cellular localization of furin between different species. Collectively, these results indicate that the unusual role of cathepsin proteases in the life cycle of bat-borne viruses is not due to the lack of active furin-like proteases in these natural reservoir species; however, differences may exist between furin proteases present in fruit bats compared to furins in other mammalian species, and these differences may impact protease usage for viral glycoprotein processing.


title: Rapid surveillance of COVID-19 in the United States using a prospective space-time scan statistic: Detecting and evaluating emerging clusters

text: Abstract Coronavirus disease 2019 (COVID-19) was first identified in Wuhan, China in December 2019, and is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 is a pandemic with an estimated death rate between 1% and 5%; and an estimated R 0 between 2.2 and 6.7 according to various sources. As of March 28th, 2020, there were over 649,000 confirmed cases and 30,249 total deaths, globally. In the United States, there were over 115,500 cases and 1891 deaths and this number is likely to increase rapidly. It is critical to detect clusters of COVID-19 to better allocate resources and improve decision-making as the outbreaks continue to grow. Using daily case data at the county level provided by Johns Hopkins University, we conducted a prospective spatial-temporal analysis with SaTScan. We detect statistically significant space-time clusters of COVID-19 at the county level in the U.S. between January 22nd-March 9th, 2020, and January 22nd-March 27th, 2020. The space-time prospective scan statistic detected “active” and emerging clusters that are present at the end of our study periods – notably, 18 more clusters were detected when adding the updated case data. These timely results can inform public health officials and decision makers about where to improve the allocation of resources, testing sites; also, where to implement stricter quarantines and travel bans. As more data becomes available, the statistic can be rerun to support timely surveillance of COVID-19, demonstrated here. Our research is the first geographic study that utilizes space-time statistics to monitor COVID-19 in the U.S.


title: Institutional Collective Action During Covid‐19: Lessons in local economic development

text: At this point, little is known about local government responses to the economic crisis caused by COVID‐19. This crisis is happening on Main Streets around the nation. This article examines how some local governments are taking collective action in partnership with others as well as organizations at the local and regional levels. What is unique is that collective action is rare as it relates to traditional economic development practices, yet it is occurring and leading to the offerings of multi‐institutional grants and low interest loans. However, some newer supply‐ and demand‐side actions are the result of a lack of resources and need for expediency. Practitioners can learn about these collaborative economic development actions other governments are taking, and how these partnerships can stabilize their local economies. This article is protected by copyright. All rights reserved.


title: Is the ferret a suitable species for studying perinatal brain injury?

text: Complications of prematurity often disrupt normal brain development and/or cause direct damage to the developing brain, resulting in poor neurodevelopmental outcomes. Physiologically relevant animal models of perinatal brain injury can advance our understanding of these influences and thereby provide opportunities to develop therapies and improve long-term outcomes. While there are advantages to currently available small animal models, there are also significant drawbacks that have limited translation of research findings to humans. Large animal models such as newborn pig, sheep and nonhuman primates have complex brain development more similar to humans, but these animals are expensive, and developmental testing of sheep and piglets is limited. Ferrets (Mustela putorius furo) are born lissencephalic and undergo postnatal cortical folding to form complex gyrencephalic brains. This review examines whether ferrets might provide a novel intermediate animal model of neonatal brain disease that has the benefit of a gyrified, altricial brain in a small animal. It summarizes attributes of ferret brain growth and development that make it an appealing animal in which to model perinatal brain injury. We postulate that because of their innate characteristics, ferrets have great potential in neonatal neurodevelopmental studies.


Issues for conducting meta-analyses in COVID-19. Commentary on “Prevalence and severity of corona virus disease 2019 (COVID-19): A systematic review and meta-analysis’’


title: Host Determinants of MERS-CoV Transmission and Pathogenesis

text: Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic pathogen that causes respiratory infection in humans, ranging from asymptomatic to severe pneumonia. In dromedary camels, the virus only causes a mild infection but it spreads efficiently between animals. Differences in the behavior of the virus observed between individuals, as well as between humans and dromedary camels, highlight the role of host factors in MERS-CoV pathogenesis and transmission. One of these host factors, the MERS-CoV receptor dipeptidyl peptidase-4 (DPP4), may be a critical determinant because it is variably expressed in MERS-CoV-susceptible species as well as in humans. This could partially explain inter- and intraspecies differences in the tropism, pathogenesis, and transmissibility of MERS-CoV. In this review, we explore the role of DPP4 and other host factors in MERS-CoV transmission and pathogenesis—such as sialic acids, host proteases, and interferons. Further characterization of these host determinants may potentially offer novel insights to develop intervention strategies to tackle ongoing outbreaks.


title: Assessment of Proton-Coupled Conformational Dynamics of SARS and MERS Coronavirus Papain-like Proteases: Implication for Designing Broad-Spectrum Antiviral Inhibitors

text: Broad-spectrum antiviral drugs are urgently needed to stop the COVID-19 pandemic and prevent future ones. The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is related to SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV), which have caused the previous outbreaks. The papain-like protease (PLpro) is an attractive drug target due to its essential roles in the viral life cycle. As a cysteine protease, PLpro is rich in cysteines and histidines and their protonation/deprotonation modulates catalysis and conformational plasticity. Here we report the pK(a) calculations and assessment of the proton-coupled conformational dynamics of SARS-CoV-2 in comparison to SARS-CoV and MERS-CoV PLpros using a newly developed GPU-accelerated implicit-solvent continuous constant pH molecular dynamics method with an asynchronous replica-exchange scheme. The calculated pK(a)’s support the catalytic roles of the Cys-His-Asp triad. We also found that several residues can switch protonation states at physiological pH, among which is C270/271 located on the flexible blocking loop 2 (BL2) of SARS-CoV-2/CoV PLpro. Simulations revealed that the BL2 conformational dynamics is coupled to the titration of C271/270, in agreement with the crystal structures of SARS-CoV-2 PLpro. Simulations also revealed that BL2 in MERS-CoV PLpro is very flexible, sampling both open and closed states despite the lack of an analogous cysteine. Our work provides a starting point for more detailed mechanistic studies to assist structure-based design of broad-spectrum inhibitors against CoV PLpros.


title: Epidemiology of diarrhoea caused by rotavirus and Escherichia coli in lambs in Kashmir valley, India

text: Abstract One hundred and twenty-nine faecal samples, collected over a period of 1 year, from 96 diarrhoeic and 33 non-diarrhoeic lambs aged between 0 and 3 months were examined for presence of rotavirus and Escherichia coli (E. coli). Group A rotavirus was detected in 24 (25%) of diarrhoeic lambs using sandwich enzyme linked immunosorbent assay (ELISA) and ribonucleic acid-polyacrylamide gel electrophoresis (RNA-PAGE). Statistically no significant relation was found between rotavirus infection and age of the lambs. The prevalence of group A rotavirus was more related to meterological changes than age of the lambs as the number of diarrhoeic lambs with rotavirus infection was found to increase in spring months during which temperature and humidity ranged between 7.34 and 28.9°C and 34.28 and 82.58%, respectively. The migration pattern of ovine rotavirus RNA in PAGE was typical of mammalian group A rotaviruses. O25, O26, O30, O43, O75, O76, O102, O113, O132, O153 and O157 E. coli serogroups were isolated from diarrhoeic lambs positive for rotavirus infection while as O8, O20, O21, O26, O39, O43, O45, O69, O75, O82, O104, O107, O113, O120, O127, O139, O141, O143, O153 and O157 serogroups of E. coli were isolated from diarrhoeic lambs without rotavirus infection. O8, O21, O43, O82, O104, O113, O120, O127, O132 and O139 serogroups, recovered from diarrhoeic faecal samples with or without rotavirus, were positive for congo red dye binding activity. O88, O113, O157 and O168 serogroups were isolated from non-diarrhoeic faecal samples out of which O88 and O168 were congo red positive. None of the lambs without diarrhoea carried rotavirus infection. Group B rotavirus infection commonly reported in diarrhoeic lambs outside India was not detected in any of the faecal samples screened.


title: CD147 overexpression on synoviocytes in rheumatoid arthritis enhances matrix metalloproteinase production and invasiveness of synoviocytes

text: Macrophage-like synoviocytes and fibroblast-like synoviocytes (FLS) are known as the most active cells of rheumatoid arthritis (RA) and are close to the articular cartilage in a position enabling them to invade the cartilage. Macrophage-like synoviocytes and FLS expression of matrix metalloproteinases (MMPs) and their interaction has aroused great interest. The present article studied the expression of CD147, also called extracellular matrix metalloproteinase inducer, on monocytes/macrophages and FLS from RA patients and its potential role in enhancing MMPs and the invasiveness of synoviocytes. Expression of CD147 on FLS derived from RA patients and from osteoarthritis patients, and expression of CD147 on monocytes/macrophages from rheumatic synovial fluid and healthy peripheral blood were analyzed by flow cytometry. The levels of CD147, MMP-2 and MMP-9 mRNA in FLS were detected by RT-PCR. The role of CD147 in MMP production and the cells' invasiveness in vitro were studied by the co-culture of FLS with the human THP-1 cell line or monocytes/macrophages, by gel zymography and by invasion assay. The results showed that the expression of CD147 was higher on RA FLS than on osteoarthritis FLS and was higher on monocytes/macrophages from rheumatic synovial fluid than on monocytes/macrophages from healthy peripheral blood. RT-PCR showed that the expressions of CD147, MMP-2 and MMP-9 mRNA was higher in RA FLS than in osteoarthritis FLS. A significantly elevated secretion and activation of MMP-2 and MMP-9 were observed in RA FLS co-cultured with differentiated THP-1 cells or RA synovial monocytes/macrophages, compared with those co-cultured with undifferentiated THP-1 cells or healthy control peripheral blood monocytes. Invasion assays showed an increased number of invading cells in the co-cultured RA FLS with differentiated THP-1 cells or RA synovial monocytes/macrophages. CD147 antagonistic peptide inhibited the MMP production and the invasive potential. Our studies demonstrated that the CD147 overexpression on monocytes/macrophages and FLS in RA patients may be responsible for the enhanced MMP secretion and activation and for the invasiveness of synoviocytes. These findings suggest that CD147 may be one of the important factors in progressive joint destruction of RA and that CD147 may be a potential therapeutic target in RA treatment.


title: Synthesis of thiocarbohydrazide and carbohydrazide derivatives as possible biologically active agents

text: ABSTRACT: A series of new β-isatin aldehyde-N,N′-thiocarbohydrazone, bis-β-isatin thiocarbohydrazones, bis-β-isatin carbohydrazones was synthesized by condensation of 5-substituted isatin with thiocarbohydrazide or carbohydrazide. The chemical structures of the newly synthesized compounds were confirmed by FT-IR, (1)H NMR, and mass spectral analysis. The synthesized compounds were evaluated for in vitro antiviral activity against various strains of DNA and RNA viruses, but exhibited moderate antiviral activity compared with the reference compounds. Among all the compounds 6c exhibited the highest chemoprevention activity in a two-stage mouse-skin carcinogenesis test. GRAPHICAL ABSTRACT: [Image: see text]


title: Non-lytic antibiotic treatment in community-acquired pneumococcal pneumonia does not attenuate inflammation: the PRISTINE trial

text: BACKGROUND: The inflammatory response in pneumococcal infection is primarily driven by immunoreactive bacterial cell wall components [lipoteichoic acid (LTA)]. An acute release of these components occurs when pneumococcal infection is treated with β-lactam antibiotics. OBJECTIVES: We hypothesized that non-lytic rifampicin compared with lytic β-lactam antibiotic treatment would attenuate the inflammatory response in patients with pneumococcal pneumonia. METHODS: In the PRISTINE (Pneumonia treated with RIfampicin aTtenuates INflammation) trial, a randomized, therapeutic controlled, exploratory study in patients with community-acquired pneumococcal pneumonia, we looked at LTA release and inflammatory and clinical response during treatment with both rifampicin and β-lactam compared with treatment with β-lactam antibiotics only. The trial is registered in the Dutch trial registry, number NTR3751 (European Clinical Trials Database number 2012-003067-22). RESULTS: Forty-one patients with community-acquired pneumonia were included; 17 of them had pneumococcal pneumonia. LTA release, LTA-mediated inflammatory responses, clinical outcomes, inflammatory biomarkers and transcription profiles were not different between treatment groups. CONCLUSIONS: The PRISTINE study demonstrated the feasibility of adding rifampicin to β-lactam antibiotics in the treatment of community-acquired pneumococcal pneumonia, but, despite solid in vitro and experimental animal research evidence, failed to demonstrate a difference in plasma LTA concentrations and subsequent inflammatory and clinical responses. Most likely, an inhibitory effect of human plasma contributes to the low immune response in these patients. In addition, LTA plasma concentration could be too low to mount a response via Toll-like receptor 2 in vitro, but may nonetheless have an effect in vivo.


title: Diagnostic Features of Clinical Neurologic Feline Infectious Peritonitis

text: Feline infectious peritonitis (FIP) is a fatal Arthus‐type immune response of cats to infection with FIP virus, a mutant of the ubiquitous feline enteric coronavirus (FECV). The disease may occur systemically or in any single organ system, and primary neurologic disease is a common subset of such manifestations. We examined 16 domestic cats with clinical neurologic FIP and 8 control cats with nonneurologic FIP, with the intention of identifying the ante‐and postmortem diagnostic tests that most contribute to accurate diagnosis. Of the 16 cats with neurologic FIP, 15 were less than 2 years of age and all 16 originated from large multiplecat households. The most useful antemortem indicators of disease were positive anti‐coronavirus IgG titer in cerebrospinal fluid, high serum total protein concentration, and findings on magnetic resonance imaging suggesting periventricular contrast enhancement, ventricular dilatation, and hydrocephalus. Postmortem diagnosis was facilitated by FIP monoclonal antibody staining of affected tissue and coronavirus‐specific polymerase chain reaction. Most cats with neurologic and ocular forms of FIP had patchy, focal lesions, suggesting that recently developed technologies described in this report may be useful for evaluation of cats with suspected FIP.


title: Managing care for patients with sinonasal and anterior skull base cancers during the COVID‐19 pandemic

text: The SARS‐CoV‐2 pandemic has rapidly transformed health care delivery around the globe. Because of the heavy impact of COVID‐19 spread, cancer treatments have necessarily been de‐prioritized, thus exposing patients to increased risk of morbidity and mortality due to delayed care. In this scenario, cancer specialists need to assess critical oncology patients case by case to carefully balance risk vs benefit in treating tumors and preventing SARS‐CoV‐2 infection. Here, we report early insights into how the management of patients with sinonasal and anterior skull base cancer might be affected by the COVID‐19 pandemic. We provide recommendations for preoperative tests, indications for immediate care vs possible delayed treatment, and warnings relating to dural resection and intracranial dissection, given the potential neurotropism of SARS‐CoV2 and practical suggestions for managing cancer care in a period of limited resources. We also postulate some thoughts on the promising role of telemedicine in multidisciplinary case discussions and posttreatment surveillance.


title: Enhanced enterovirus 71 virus‐like particle yield from a new baculovirus design

text: Enterovirus 71 (EV71) is responsible for the outbreaks of hand‐foot‐and‐mouth disease in the Asia‐Pacific region. To produce the virus‐like particle (VLP) vaccine, we previously constructed recombinant baculoviruses to co‐express EV71 P1 polypeptide and 3CD protease using the Bac‐to‐Bac(®) vector system. The recombinant baculoviruses resulted in P1 cleavage by 3CD and subsequent VLP assembly in infected insect cells, but caused either low VLP yield or excessive VLP degradation. To tackle the problems, here we explored various expression cassette designs and flashBAC GOLD™ vector system which was deficient in v‐cath and chiA genes. We found that the recombinant baculovirus constructed using the flashBAC GOLD™ system was insufficient to improve the EV71 VLP yield. Nonetheless, BacF‐P1‐C3CD, a recombinant baculovirus constructed using the flashBAC GOLD(TM) system to express P1 under the polh promoter and 3CD under the CMV promoter, dramatically improved the VLP yield while alleviating the VLP degradation. Infection of High Five(TM) cells with BacF‐P1‐C3CD enhanced the total and extracellular VLP yield to ≈268 and ≈171 mg/L, respectively, which enabled the release of abundant VLP into the supernatant and simplified the downstream purification. Intramuscular immunization of mice with 5 μg purified VLP induced cross‐protective humoral responses and conferred protection against lethal virus challenge. Given the significantly improved extracellular VLP yield (≈171 mg/L) and the potent immunogenicity conferred by 5 μg VLP, one liter High Five(TM) culture produced ≈12,000 doses of purified vaccine, thus rendering the EV71 VLP vaccine economically viable and able to compete with inactivated virus vaccines. Biotechnol. Bioeng. 2015;112: 2005–2015. © 2015 Wiley Periodicals, Inc.


title: Development and implementation of influenza a virus subtyping and detection of genotypic resistance to neuraminidase inhibitors

text: Influenza virus hemagglutinin and neuraminidase, surface glycoproteins with an essential role in viral pathogenesis, are important antigen determinants and essential markers for epidemiological surveillance. Neuraminidase is also a suitable target for designing antiviral drugs. The introduction into clinical practice of neuraminidase inhibitors and the development of random point mutations have increased the emergence of drug‐resistant viruses. A universal RT nested PCR‐based system has been developed for subtyping H1, H3, N1 and N2, in influenza A viruses of human or animal origin. The subsequent sequencing and analysis of the hemagglutinin and neuraminidase templates reveal antigenic and receptor binding changes in the HA1 subunit and mutations of clinical relevance concerning resistance to neuraminidase inhibitors. The specificity and sensitivity of the method were evaluated using 113 influenza A isolates, 105 influenza A positive respiratory samples obtained from patients and 29 prototype strains of both human and animal origin. The resulting analytical sensitivity of the subtyping techniques is one to at least 100 molecules of cloned DNA product in a final reaction volume of 50 µl. In the course of implementing the method, two H1N1 isolates with the H274Y mutation in the neuraminidase segment have been detected and their molecular features analyzed. The emergence of influenza virus resistance makes the neuraminidase genetic characterization and surveillance activities to detect antiviral resistance necessary. J. Med. Virol. 82: 843–853, 2010. © 2010 Wiley‐Liss, Inc.