The Transformation of Enterovirus Replication Structures: a Three-Dimensional Study of Single- and Double-Membrane Compartments

All positive-strand RNA viruses induce membrane structures in their host cells which are thought to serve as suitable microenvironments for viral RNA synthesis. The structures induced by enteroviruses, which are members of the family Picornaviridae, have so far been described as either single- or double-membrane vesicles (DMVs). Aside from the number of delimiting membranes, their exact architecture has also remained elusive due to the limitations of conventional electron microscopy. In this study, we used electron tomography (ET) to solve the three-dimensional (3-D) ultrastructure of these compartments. At different time points postinfection, coxsackievirus B3-infected cells were high-pressure frozen and freeze-substituted for ET analysis. The tomograms showed that during the exponential phase of viral RNA synthesis, closed smooth single-membrane tubules constituted the predominant virus-induced membrane structure, with a minor proportion of DMVs that were either closed or connected to the cytosol in a vase-like configuration. As infection progressed, the DMV number steadily increased, while the tubular single-membrane structures gradually disappeared. Late in infection, complex multilamellar structures, previously unreported, became apparent in the cytoplasm. Serial tomography disclosed that their basic unit is a DMV, which is enwrapped by one or multiple cisternae. ET also revealed striking intermediate structures that strongly support the conversion of single-membrane tubules into double-membrane and multilamellar structures by a process of membrane apposition, enwrapping, and fusion. Collectively, our work unravels the sequential appearance of distinct enterovirus-induced replication structures, elucidates their detailed 3-D architecture, and provides the basis for a model for their transformation during the course of infection

Identification of regulatory variants associated with genetic susceptibility to meningococcal disease.

Non-coding genetic variants play an important role in driving susceptibility to complex diseases but their characterization remains challenging. Here, we employed a novel approach to interrogate the genetic risk of such polymorphisms in a more systematic way by targeting specific regulatory regions relevant for the phenotype studied. We applied this method to meningococcal disease susceptibility, using the DNA binding pattern of RELA - a NF-kB subunit, master regulator of the response to infection - under bacterial stimuli in nasopharyngeal epithelial cells. We designed a custom panel to cover these RELA binding sites and used it for targeted sequencing in cases and controls. Variant calling and association analysis were performed followed by validation of candidate polymorphisms by genotyping in three independent cohorts. We identified two new polymorphisms, rs4823231 and rs11913168, showing signs of association with meningococcal disease susceptibility. In addition, using our genomic data as well as publicly available resources, we found evidences for these SNPs to have potential regulatory effects on ATXN10 and LIF genes respectively. The variants and related candidate genes are relevant for infectious diseases and may have important contribution for meningococcal disease pathology. Finally, we described a novel genetic association approach that could be applied to other phenotypes

Plasma lipid profiles discriminate bacterial from viral infection in febrile children

Fever is the most common reason that children present to Emergency Departments. Clinical signs and symptoms suggestive of bacterial infection are often non-specific, and there is no definitive test for the accurate diagnosis of infection. The 'omics' approaches to identifying biomarkers from the host-response to bacterial infection are promising. In this study, lipidomic analysis was carried out with plasma samples obtained from febrile children with confirmed bacterial infection (n = 20) and confirmed viral infection (n = 20). We show for the first time that bacterial and viral infection produces distinct profile in the host lipidome. Some species of glycerophosphoinositol, sphingomyelin, lysophosphatidylcholine and cholesterol sulfate were higher in the confirmed virus infected group, while some species of fatty acids, glycerophosphocholine, glycerophosphoserine, lactosylceramide and bilirubin were lower in the confirmed virus infected group when compared with confirmed bacterial infected group. A combination of three lipids achieved an area under the receiver operating characteristic (ROC) curve of 0.911 (95% CI 0.81 to 0.98). This pilot study demonstrates the potential of metabolic biomarkers to assist clinicians in distinguishing bacterial from viral infection in febrile children, to facilitate effective clinical management and to the limit inappropriate use of antibiotics

Plasma lipid profiles discriminate bacterial from viral infection in febrile children

Fever is the most common reason that children present to Emergency Departments. Clinical signs and symptoms suggestive of bacterial infection are often non-specific, and there is no definitive test for the accurate diagnosis of infection. The 'omics' approaches to identifying biomarkers from the host-response to bacterial infection are promising. In this study, lipidomic analysis was carried out with plasma samples obtained from febrile children with confirmed bacterial infection (n = 20) and confirmed viral infection (n = 20). We show for the first time that bacterial and viral infection produces distinct profile in the host lipidome. Some species of glycerophosphoinositol, sphingomyelin, lysophosphatidylcholine and cholesterol sulfate were higher in the confirmed virus infected group, while some species of fatty acids, glycerophosphocholine, glycerophosphoserine, lactosylceramide and bilirubin were lower in the confirmed virus infected group when compared with confirmed bacterial infected group. A combination of three lipids achieved an area under the receiver operating characteristic (ROC) curve of 0.911 (95% CI 0.81 to 0.98). This pilot study demonstrates the potential of metabolic biomarkers to assist clinicians in distinguishing bacterial from viral infection in febrile children, to facilitate effective clinical management and to the limit inappropriate use of antibiotics

Plasma lipid profiles discriminate bacterial from viral infection in febrile children

Fever is the most common reason that children present to Emergency Departments. Clinical signs and symptoms suggestive of bacterial infection ar

Richtlijnen

Multidisciplinaire richtlijnen zijn niet meer weg te denken uit de gezondheidszorg. Richtlijnen ondersteunen evidence-based werken, waarbij de beste beschikbare wetenschappelijke literatuur wordt gecombineerd met de expertise van de professional en de expertise en voorkeuren van patiënten. Richtlijnen staan boven in de piramide van evidence (nog boven systematische reviews), maar dit gaat alleen op als het een richtlijn van goede kwaliteit betreft. Om de methodologische kwaliteit van een richtlijn te controleren, wordt de internationale standaard AGREE-II gebruikt. Dit gevalideerde instrument beoordeelt de kwaliteit van een richtlijn met 23 items in zes domeinen. Werken volgens richtlijnen is een middel om goede patiëntenzorg te kunnen leveren. De verpleegkundige en verpleegkundig specialist moeten echter aandacht hebben voor de kwaliteit van de richtlijn om de kwaliteit van de aanbevelingen op waarde te kunnen schatten

Unlock the doors: aggressive behaviour and seclusion on close psychiatric wards

Seclusion of patients on psychiatric wards is a controversial intervention with serious safety issues for patients. Nurses have an important role in the decision to use seclusion when encountering aggressive behaviour. The aim of this thesis is to improve knowledge about the influence of nurses on the incidence of aggressive behaviour and the use of seclusion, in order to use that knowledge to prevent such events in the future. Patients and nurses share views on the factual cause of aggressive behaviour, but their perception of the severity differ. The attitude of nurses towards seclusion changed over the last decades from a therapeutic paradigm to a safety paradigm. In our study, teams with majority of male nurses were associated with less seclusion and aggressive behaviour. Teams with high scores on personality trait openness were associated with less seclusion and teams with high scores on personality trait extraversion were associated with patients’ aggression. There are no golden solutions to improve safety of psychiatric wards, but increased attention of development of nursing skills seems as an important step forward. By providing optimal nursing care, vulnerable patients on psychiatric wards receive the care they deserve

Verpleegkundige factoren bij het voorkomen van separatie: een prognostische observationele studie

INTRODUCTIE Separatie is een veel gebruikte, maar controversiële interventie, zonder bewezen therapeutisch effect. (1) Patiënten rapporteren negatieve ervaringen na separatie. (2) Verschillende studies laten staf - en afdelingskenmerken zien die bijdragen aan separatie. Vanwege niet eenduidige uitkomsten is er behoefte aan een prospectief onderzoek naar het verband tussen deze determinanten met separatie. DOELSTELLING Een prospectief onderzoek naar (verpleegkundige) staf kenmerken die verband houden met separatie bij patiënten in de acute psychiatrie