A school-based program to prevent depressive symptoms and strengthen well-being among pre-vocational students (Happy Lessons):protocol for a cluster randomized controlled trial and implementation study

BACKGROUND: Depression is one of the leading causes of illness and disability among young people. In the Netherlands, one in twelve Dutch adolescents has experienced depression in the last 12 months. Pre-vocational students are at higher risk for elevated depressive symptoms. Effective interventions, especially for this risk group, are therefore needed to prevent the onset of depression or mitigate the adverse long-term effects of depression. The aim of this study is to examine the effectiveness and implementation of a school-based program Happy Lessons (HL), that aims to prevent depression and promote well-being among pre-vocational students. METHODS: A cluster randomized controlled trial (RCT) with students randomized to HL or to care as usual will be conducted. Pre-vocational students in their first or second year (aged 12 to 14) will participate in the study. Subjects in both conditions will complete assessments at baseline and at 3- and 6-months follow-up. The primary outcome will be depressive symptoms using the Center for Epidemiologic Studies Depression Scale (CES-D) at 6-months follow-up. Secondary outcomes are well-being using the Warwick-Edinburgh Mental Well-Being Scale (WEMWBS) and life satisfaction (Cantril Ladder) measured at 6-months follow-up. Alongside the trial, an implementation study will be conducted to evaluate the implementation of HL, using both quantitative and qualitative methods (interviews, survey, and classroom observations). DISCUSSION: The results from both the RCT and implementation study will contribute to the limited evidence base on effective school-based interventions for the prevention of depression and promotion of well-being among pre-vocational adolescents. In addition, insights from the implementation study will aid identifying factors relevant for optimizing the future implementation and scale-up of HL to other schools and contexts. TRIAL REGISTRATION: This study was registered on 20 September 2021 in the Dutch Trial Register (NL9732). SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12889-021-12321-3

Het mentaal welbevinden van jongeren in het praktijkonderwijs en cluster 4-onderwijs: Cijfers uit het EXPLORE-onderzoek 2019

Het EXPLORE-onderzoek is een landelijk onderzoek naar de omvang van middelengebruik onder jongeren in het praktijkonderwijs, voortgezet speciaal onderwijs (cluster 3 en 4), residentiële jeugdzorg en justitiële jeugdinrichtingen. Het doel van het EXPLORE-onderzoek is gegevens te leveren ter onderbouwing en ontwikkeling van een effectieve preventieve aanpak van (problematisch) middelengebruik onder kwetsbare jongeren. Naast middelengebruik zijn er op het praktijkonderwijs en cluster 4- onderwijs een aantal vragen gesteld over het mentaal welbevinden

Het mentaal welbevinden van jongeren in het praktijkonderwijs en cluster 4-onderwijs: Cijfers uit het EXPLORE-onderzoek 2019

Het EXPLORE-onderzoek is een landelijk onderzoek naar de omvang van middelengebruik onder jongeren in het praktijkonderwijs, voortgezet speciaal onderwijs (cluster 3 en 4), residentiële jeugdzorg en justitiële jeugdinrichtingen. Het doel van het EXPLORE-onderzoek is gegevens te leveren ter onderbouwing en ontwikkeling van een effectieve preventieve aanpak van (problematisch) middelengebruik onder kwetsbare jongeren. Naast middelengebruik zijn er op het praktijkonderwijs en cluster 4- onderwijs een aantal vragen gesteld over het mentaal welbevinden

Human 3-D tissue models in radiation biology: current status and future perspectives

In this review, we discuss the use of a variety of 3‐D models (particularly 3‐D skin, lung, breast and endothelial) in radiobiological research and highlight the differences in responses compared to 2‐D culturing conditions (monolayers). We review the characteristics of existing 3‐D models and aim to point out the substantial advantages 3‐D cultures provide for modern radiobiology. In particular, they may facilitate the shift from the classical DNA damage and repair studies mainly carried out in monolayer cultures to the investigation of more generalized responses through pathway analysis and a system biology approach. 3‐D models are expected to be very informative for investigations on radiotherapy responses in addressing the low dose risk. However, the 3‐D model systems are not as easy to propagate and standardize as monolayer cultures. Therefore, we discuss the problems and limitations of 3‐D models and propose ways to overcome some of the problems

Adaptive antibody diversification through-linked glycosylation of the immunoglobulin variable region

A hallmark of B-cell immunity is the generation of a diverse repertoire of antibodies from a limited set of germline V(D)J genes. This repertoire is usually defined in terms of amino acid composition. However, variable domains may also acquire-linked glycans, a process conditional on the introduction of consensus amino acid motifs (-glycosylation sites) during somatic hypermutation. High levels of variable domain glycans have been associated with autoantibodies in rheumatoid arthritis, as well as certain follicular lymphomas. However, the role of these glycans in the humoral immune response remains poorly understood. Interestingly, studies have reported both positive and negative effects on antibody affinity. Our aim was to elucidate the role of variable domain glycans during antigen-specific antibody responses. By analyzing B-cell repertoires by next-generation sequencing, we demonstrate that-glycosylation sites are introduced at positions in which glycans can affect antigen binding as a result of a specific clustering of progenitor glycosylation sites in the germline sequences of variable domain genes. By analyzing multiple human monoclonal and polyclonal (auto)antibody responses, we subsequently show that this process is subject to selection during antigen-specific antibody responses, skewed toward IgG4, and positively contributes to antigen binding. Together, these results highlight a physiological role for variable domain glycosylation as an additional layer of antibody diversification that modulates antigen binding.status: publishe

Host phospholipid peroxidation fuels ExoU-dependent cell necrosis and supports Pseudomonas aeruginosaPseudomonas\ aeruginosa-driven pathology

International audienceRegulated cell necrosis supports immune and anti-infectious strategies of the body; however, dysregulation of these processes drives pathological organ damage. $Pseudomonas\ aeruginosa$ expresses a phospholipase, ExoU that triggers pathological host cell necrosis through a poorly characterized pathway. Here, we investigated the molecular and cellular mechanisms of ExoU-mediated necrosis. We show that cellular peroxidised phospholipids enhance ExoU phospholipase activity, which drives necrosis of immune and non-immune cells. Conversely, both the endogenous lipid peroxidation regulator GPX4 and the pharmacological inhibition of lipid peroxidation delay ExoU-dependent cell necrosis and improve bacterial elimination in vitro and in vivo . Our findings also pertain to the ExoU-related phospholipase from the bacterial pathogen $Burkholderia\ thailandensis$ , suggesting that exploitation of peroxidised phospholipids might be a conserved virulence mechanism among various microbial phospholipases. Overall, our results identify an original lipid peroxidation-based virulence mechanism as a strong contributor of microbial phospholipase-driven pathology

N-linked glycosylation of the immunoglobulin variable region

N-glycosylation sites are introduced at positions in which glycans can affect antigen binding as a result of a specific clustering of progenitor glycosylation sites in the germline sequences of variable domain genes. By analyzing multiple human monoclonal and polyclonal (auto)antibody responses, we subsequently show that this process is subject to selection during antigen-specific antibody responses, skewed toward IgG4, and positively contributes to antigen binding. Together, these results highlight a physiological role for variable domain glycosylation as an additional layer of antibody diversification that modulates antigen bindin

MmpL Genes Are Associated with Mycolic Acid Metabolism in Mycobacteria and Corynebacteria

Mycolic acids are vital components of the cell wall of the tubercle bacillus Mycobacterium tuberculosis and are required for viability and virulence. While mycolic acid biosynthesis is studied extensively, components involved in mycolate transport remain unidentified. We investigated the role of large membrane proteins encoded by mmpL genes in mycolic acid transport in mycobacteria and the related corynebacteria. MmpL3 was found to be essential in mycobacteria and conditional depletion of MmpL3 in Mycobacterium smegmatis resulted in loss of cell wall mycolylation, and of the cell wall-associated glycolipid, trehalose dimycolate. In parallel, an accumulation of trehalose monomycolate (TMM) was observed, suggesting that mycolic acids were transported as TMM. In contrast to mycobacteria, we found redundancy in the role of two mmpL genes, in Corynebacterium glutamicum; a complete loss of trehalose-associated and cell wall bound corynomycolates was observed in an NCgl0228-NCgl2769 double mutant, but not in individual single mutants. Our studies highlight the role of mmpL genes in mycolic acid metabolism and identify potential new targets for anti-TB drug development

Pseudomonas aeruginosa infection reveals a Caspase-1-dependent neutrophil pyroptosis pathway that restrains damaging Histone release

Abstract Neutrophils mediate essential immune and microbicidal processes. Consequently, to counteract neutrophil attack, pathogens have developed various virulence strategies. Here, we showed that Pseudomonas aeruginosa ( P. aeruginosa ) phospholipase ExoU drives pathological NETosis in neutrophils. Surprisingly, inhibition of ExoU activity uncovered a fully functional Caspase-1-driven pyroptosis pathway in neutrophils. Mechanistically, activated NLRC4 inflammasome promoted Caspase-1-dependent Gasdermin-D activation, IL-1β cytokine release and neutrophil pyroptosis. Whereas both pyroptotic and netotic neutrophils released alarmins, only NETosis liberated the destructive DAMPs Histones, which exacerbated Pseudomonas -induced mouse lethality. To the contrary, subcortical actin allowed pyroptotic neutrophils to physically limit poisonous inflammation by keeping Histones intracellularly. Finally, mouse models of infection highlighted that both NETosis and neutrophil Caspase-1 contributed to P. aeruginosa spreading. Overall, we established the host deleterious consequences of Pseudomonas -induced-NETosis but also uncovered an unsuspected ability of neutrophils to undergo Caspase-1-dependent pyroptosis, a process where neutrophils exhibit a self-regulatory function that limit Histone release. Graphical abstract P.aeruginosa ExoU (right) triggers phospholipid degradation and subsequent neutrophil lysis that associates to NETosis through F-Actin collapse/degradation and PAD4-dependent DNA decondensation. P.aeruginosa ExoU- (left) triggers NLRC4-dependent pyroptosis in neutrophils, which leads to PAD4- depndent DNA decondensation but not expulsion due to a still function subcortical F- Actin network. Created with Biorender.com