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Uit onderzoek blijkt dat samenredzaamheid, het gevoel dat gemeenschapsleden door gezamenlijke inspanning hun problemen kunnen oplossen, belangrijk is voor het welzijn van cliënten. De interventie Community Support focust op het versterken van de samenredzaamheid van cliënten door onder andere het sociale netwerk te betrekken en uit te breiden. Het huidige onderzoek richt zich op de ervaringen van jeugdigen, ouders/ verzorgers en personen uit het sociale netwerk met het betrekken van het sociale netwerk en de bijdrage aan netwerkversterking binnen de interventie. Hiervoor zijn duo, semigestructureerde interviews afgenomen. Uit het huidige onderzoek blijkt dat het merendeel van de participanten tevreden is over de betrokkenheid van het sociale netwerk bij de hulp. Belangrijke factoren die de betrokkenheid van het sociale netwerk bevorderen zijn: 1) het bieden van een luisterend oor, 2) het actief meedenken, 3) het betrekken van meerdere perspectieven en 4) netwerkbijeenkomsten. Belemmerende factoren zijn: 1) angst voor oordeel van de omgeving, 2) overbelasting van belangrijke steunfiguren, 3) schaamte voor de problematiek, 4) gebrek aan begrip en 5) communicatie. Daarnaast heeft het merendeel van de participanten een verandering in het sociale netwerk na de interventie opgemerkt. Echter niet alle participanten zijn tevreden met de ontvangen sociale steun na de hulp. Tot slot blijkt dat de participanten verdeeld zijn over de mate waarin zij de veranderingen in hun sociale netwerk toeschrijven aan de interventie. Belangrijke factoren die de bijdrage van de interventie aan netwerkversterking bevorderen zijn: 1) actief betrekken van het sociale netwerk en 2) sociaal contact leggen. De participanten die aangeven dat de interventie weinig heeft bijgedragen op het gebied van netwerversterking, benoemen dat de contacten met het sociale netwerk al bestonden vóór de hulp en zij zelfstandig het sociale netwerk betrekken bij de hulpverlening. Concluderend, uit het huidige onderzoek blijkt dat er geen eenduidige manier is om het sociale netwerk te betrekken bij de hulpverlening. Om deze reden is het van belang om met cliënten in gesprek te blijven gaan over de mogelijke drempels die zij ervaren en hoe deze verminderd kunnen worden. Vervolgonderzoek zou zich kunnen richten op de ervaringen van professionals met het betrekken van het sociale netwerk bij de hulpverlening, met als doel hen hierin beter te ondersteunen.

Glycomics aims to identify the structure and function of the glycome, the complete set of oligosaccharides (glycans), produced in a given cell or organism, as well as to identify genes and other factors that gover n glycosylation. This challenging endeavor requires highly robust, sensitive, and potentially automatable analytical technologies for the analysis of hundreds or thousands of glycomes in a timely manner (termed high-throughput glycomics). This review provides a historic overview as well as highlights recent developments and challenges of glycomic profi l i n g by the most prominent high-throughput glycomic approaches, with N-glycosylation analysis as the focal point. It describes the current state-of-the-art regarding levels of characterization and most widely used technologies, selected applications of high-throughput glycomics in deciphering glycosylation process in healthy and disease states, as well as future perspectives.

The ability to preserve metabolically active kidneys ex vivo for multiple days may permit reconditioning, repair and regeneration of deceased donor kidneys. However, the kidneys high metabolic demand limits its functional preservation. Current approaches focus on normothermic machine perfusion (NMP) at 37°C or hypothermic machine perfusion (HMP) at 4-8°C. At normothermia, kidneys are metabolically active butex vivopreservation is limited to hours. During hypothermia kidneys can be preserved up to 24 hours but are metabolically inactive and suffer cold-induced injury. Therefore, we revisited sub normothermic perfusion (at 25°C) as an alternative approach to preserve human kidneys in a metabolically active state for extended periods of time.In a custom-made platform that includes a cell-free perfusate enriched with TCA cycle fuels, urine recirculation, and continuous hemofiltration we perfused discarded human kidneys up to 8 days. Using spatially resolved single cell resolution isotope tracing we demonstrate active metabolism in all the different renal cell types over this period. However, beyond 4 days cell composition of nephron segments assessed with spatial lipidomics changed substantially and injury markers such as NGAL and LDH increased in the perfusate. Up to 4 days, perfused human discarded donor kidneys maintained metabolic fluxes, functional parameters and allow for reperfusion using a porcine auto transplantation model. These data underpin that extended multi-day metabolic preservation of human kidneys is achievable using a sub normothermic perfusion platform.

Supplementary Figure 1 from High-Throughput Characterization of 10 New Minor Histocompatibility Antigens by Whole Genome Association Scanning

Supplementary Tables 1-3 from High-Throughput Characterization of 10 New Minor Histocompatibility Antigens by Whole Genome Association Scanning

Patients with malignant diseases can be effectively treated with allogeneic hematopoietic stem cell transplantation (allo-SCT). Polymorphic peptides presented in HLA molecules, the so-called minor histocompatibility antigens (MiHA), play a crucial role in antitumor immunity as targets for alloreactive donor T cells. Identification of multiple MiHAs is essential to understand and manipulate the development of clinical responses after allo-SCT. In this study, CD8+ T-cell clones were isolated from leukemia patients who entered complete remission after allo-SCT, and MiHA-specific T-cell clones were efficiently selected for analysis of recognition of a panel of EBV-transformed B cells positive for the HLA restriction elements of the selected T-cell clones. One million single nucleotide polymorphisms (SNP) were determined in the panel cell lines and investigated for matching with the T-cell recognition data by whole genome association scanning (WGAs). Significant association with 12 genomic regions was found, and detailed analysis of genes located within these genomic regions revealed SNP disparities encoding polymorphic peptides in 10 cases. Differential recognition of patient-type, but not donor-type, peptides validated the identification of these MiHAs. Using tetramers, distinct populations of MiHA-specific CD8+ T cells were detected, demonstrating that our WGAs strategy allows high-throughput discovery of relevant targets in antitumor immunity after allo-SCT. Cancer Res; 70(22); 9073–83. ©2010 AACR.

Severe allergic reactions to certain types of meat following tick bites have been reported in geographic regions which are endemic with ticks. This immune response is directed to a carbohydrate antigen (galactose-alpha-1,3-galactose or alpha-Gal), which is present in glycoproteins of mammalian meats. At the moment, asparagine-linked complex carbohydrates (N-glycans) with alpha-Gal motifs in meat glycoproteins and in which cell types or tissue morphologies these alpha-Gal moieties are present in mammalian meats are still unclear. In this study, we analyzed alpha-Gal-containing N-glycans in beef, mutton, and pork tenderloin and provided for the first time the spatial distribution of these types of N-glycans in various meat samples. Terminal alpha-Gal-modified N-glycans were found to be highly abundant in all analyzed samples (55, 45, and 36% of N-glycome in beef, mutton, and pork, respectively). Visualizations of the N-glycans with alpha-Gal modification revealed that this motif was mainly present in the fibroconnective tissue. To conclude, this study contributes to a better understanding of the glycosylation biology of meat samples and provides guidance for processed meat products, in which only meat fibers are required as an ingredient (i.e., sausages or canned meat).

Analytical techniques with high sensitivity and selectivity are essential to the quantitative analysis of clinical samples. Liquid chromatography coupled to tandem mass spectrometry is the gold standard in clinical chemistry. However, tandem mass spectrometers come at high capital expenditure and maintenance costs. We recently showed that it is possible to generate very similar results using a much simpler single mass spectrometry detector by performing enhanced in-source fragmentation/annotation (EISA) combined with correlated ion monitoring. Here we provide a step-by-step protocol for optimizing the analytical conditions for EISA, so anyone properly trained in liquid chromatography-mass spectrometry can follow and apply this technique for any given analyte. We exemplify the approach by using 2-hydroxyglutarate (2-HG) which is a clinically relevant metabolite whose d-enantiomer is considered an 'oncometabolite', characteristic of cancers associated with mutated isocitrate dehydrogenases 1 or 2 (IDH1/2). We include procedures for determining quantitative robustness, and show results of these relating to the analysis of dl-2-hydroxyglutarate in cells, as well as in serum samples from patients with acute myeloid leukemia that contain the IDH1/2 mutation. This EISA-mass spectrometry protocol is a broadly applicable and low-cost approach for the quantification of small molecules that has been developed to work well for both single-quadrupole and time-of-flight mass analyzers.The tandem mass spectrometers used in clinical chemistry are expensive. This protocol describes how to generate similar results using a single mass spectrometry detector by optimizing in-source fragmentation and data analysis via correlated ion monitoring.

Background The prevalence of depression has increased among adolescents in western countries. Prevention is needed to reduce the number of adolescents who experience depression and to avoid negative consequences, including suicide. Several preventive interventions are found to be promising, especially multi-modal approaches, for example combining screening and preventive intervention. However, an important bottleneck arises during the implementation of preventive intervention. Only a small percentage of adolescents who are eligible for participation actually participate in the intervention. To ensure that more adolescents can benefit from prevention, we need to close the gap between detection and preventive intervention. We investigated the barriers and facilitators from the perspective of public health professionals in screening for depressive and suicidal symptoms and depression prevention referral in a school-based setting. Methods We conducted 13 semi-structured interviews with public health professionals, who execute screening and depression prevention referral within the Strong Teens and Resilient Minds (STORM) approach. The interviews were recorded, transcribed verbatim, and coded in several cycles using ATLAS.ti Web. Results Three main themes of barriers and facilitators emerged from the interviews, namely “professional capabilities,” “organization and collaboration,” and “beliefs about depressive and suicidal symptoms and participation in prevention”. The interviews revealed that professionals do not always feel sufficiently equipped in terms of knowledge, skills and supporting networks. Consequently, they do not always feel well able to execute the process of screening and prevention referral. In addition, a lack of knowledge and support in schools and other cooperating organizationorganizations was seen to hinder the process. Last, the beliefs of public health professionals, school staff, adolescents, and parents —especially stigma and taboo—were found to make the screening and prevention referral process more challenging. Conclusions To further improve the process of screening and prevention referral in a school-based setting, enhancing professional competence and a holding work environment for professionals, a strong collaboration and a joint approach with schools and other cooperating organizations and society wide education about depressive and suicidal symptoms and preventive intervention are suggested. Future research should determine whether these recommendations actually lead to closing the gap between detection and prevention.

What explains the different additionality of R&D subsidies through public programmes of different government levels? Does multiple programme participation induce more additionality? We deal with these questions by examining the profile of 8,280 Spanish firms with R&D projects publicly supported through subsidy programmes for the period 2007–2016. We use standard Propensity Score Matching techniques to estimate treatment effects at the firm level, and then we explore the reasons for the heterogeneity in these individual effects through the estimation of an equation for their determinants. We find that, after controlling for a large set of firm characteristics, companies with multiple programme participation show higher additionality, although the effect is lower for firms with very high support intensities. Second, regardless of the government level of the support programme, the degree of additionality is related positively to firm innovativeness and negatively to dimensions that denote a more market-oriented R&D.

Objectives Changes in protein glycosylation are a hallmark of immune-mediated diseases. Glycans are master regulators of the inflammatory response being important molecules for the discrimination between "self"/"non-self". We here explored whether lupus nephritis (LN) exhibit an altered cellular glycosylation aiming to identify a unique glycosignature that characterizes LN pathogenesis. Methods A comprehensive tissue glycomics characterization was performed in a cohort of human biopsy-proven LN clinical samples from SLE patients. A combination of advanced tissue mass spectrometry imaging (MSI); in situ glyco-characterization and ex vivo glycophenotyping of human kidney biopsies were performed to structurally map the N-glycans repertoire in LN samples. Results LN revealed a unique glycan signature characterized by an increased abundance and spatial distribution of unusual mannose-enriched glycans that are typically found in lower microorganisms; this glycosignature was specific of LN as it was not observed in other kidney diseases. Exposure of mannosylated glycans in LN was found to occur at the cell surface of kidney cells promoting an increased recognition by specific glycans-recognizing receptors, expressed by immune cells. This abnormal glyco-signature of LN was demonstrated to be due to a deficient complex N-glycosylation and a proficient O-mannosylation pathway. Moreover, levels of mannosylation detected in kidney biopsies from LN patients at diagnosis were demonstrated to predict the development of chronic kidney disease (CKD) with 93% of specificity. Conclusions Cellular mannosylation is a marker of LN, predicting the development of CKD, and thus constituting a potential glycobiomarker to be included in the diagnostic and prognostic algorithm of LN.

Conventional single-cell metabolomics approaches such as matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) generate biochemical snapshots, neglecting the inherent dynamic nature of metabolism. Here we describe a platform based on isotope tracing and MALDI-MSI that allows in-situ dynamic measurements of cell type-specific metabolism at single-cell resolution, and thus unravel cell metabolism within its tissue architecture. We applied different 13C-isotope-labeled nutrients on vibratome slices of fresh mouse kidney. MALDI-MSI at single-cell resolution (i.e. pixel size of 5 × 5 μm2) was then applied to detect metabolites and lipids from the harvested tissues. Following MALDI-MSI analysis, post-MSI-analyzed sections were stained and subsequently imaged using multiplexed immunofluorescence (IF) microscopy for cell-type identification. This platform allows to achieve single-cell resolution in situ and hence interpret cell type-specific metabolic dynamics in the context of structure and metabolism of neighboring cells.