Direct Visualization and Silver Enhancement of Ultra-Small Antibody-Bound Gold Particles on Immunolabeled Ultrathin Resin Sections

Ultra-small gold colloids bound to immunolabeled ultrathin resin sections were visualized using transmission, scanning, and scanning transmission electron microscopy (TEM, SEM, STEM). The best marker contrast is obtained in a field emission STEM (200 kV) equipped with a high-angle annular dark-field (HAADF) detector. HAADF STEM renders possible the simultaneous visualization of ultra-small gold and ultrastructural details in unstained resin sections, and an overall presentation of a labeled E. coli cell. For routine work, an enhancement step is a prerequisite for easy detection of bound marker molecules. Five different silver enhancing solutions were tested for their suitability for ultra-small gold intensification. Enhancers lacking the protective colloid gum arabic exhibit lower quality with regard to efficiency and homogeneity of enhancement. This problem can be overcome by adding gum arabic. Silver enhancement generally results in heterogeneously sized particles. This is most probably due to the heterogeneous original gold colloid probe. In general, an estimation of enhancement efficiency is associated with difficulties depending on experimental conditions and the electron microscopic imaging modes used. Only a low number of the ultra-small gold particles seems to remain unenhanced or poorly enhanced when treated with high-quality enhancers. On-section labeling of ultrathin resin sections with silver-enhanced ultra-small gold markers also offers the possibility of high-resolution immunolabeling experiments at the light microscopic level

Assessment of mRNA and microRNA Stabilization in Peripheral Human Blood for Multicenter Studies and Biobanks

In this study we evaluate the suitability of two methods of RNA conservation in blood samples, PAXgene and RNAlater, in combination with variable shipping conditions for their application in multicenter studies and biobanking. RNA yield, integrity, and purity as well as levels of selected mRNA and microRNA species were analyzed in peripheral human blood samples stabilized by PAXgene or RNAlater and shipped on dry ice or at ambient temperatures from the study centers to the central analysis laboratory. Both examined systems were clearly appropriate for RNA stabilization in human blood independently of the shipping conditions. The isolated RNA is characterized by good quantity and quality and well suited for downstream applications like quantitative RT-PCR analysis of mRNA and microRNA. Superior yield and integrity values were received using RNAlater. It would be reasonable to consider the production and approval of blood collection tubes prefilled with RNAlater to facilitate the use of this excellent RNA stabilization system in large studies

SKA2 regulated hyperactive secretory autophagy drives neuroinflammation-induced neurodegeneration

High levels of proinflammatory cytokines induce neurotoxicity and catalyze inflammation-driven neurodegeneration, but the specific release mechanisms from microglia remain elusive. Here we show that secretory autophagy (SA), a non-lytic modality of autophagy for secretion of vesicular cargo, regulates neuroinflammation-mediated neurodegeneration via SKA2 and FKBP5 signaling. SKA2 inhibits SA-dependent IL-1β release by counteracting FKBP5 function. Hippocampal Ska2 knockdown in male mice hyperactivates SA resulting in neuroinflammation, subsequent neurodegeneration and complete hippocampal atrophy within six weeks. The hyperactivation of SA increases IL-1β release, contributing to an inflammatory feed-forward vicious cycle including NLRP3-inflammasome activation and Gasdermin D-mediated neurotoxicity, which ultimately drives neurodegeneration. Results from protein expression and co-immunoprecipitation analyses of male and female postmortem human brains demonstrate that SA is hyperactivated in Alzheimer's disease. Overall, our findings suggest that SKA2-regulated, hyperactive SA facilitates neuroinflammation and is linked to Alzheimer's disease, providing mechanistic insight into the biology of neuroinflammation

Zentrale Archivierung und verteilte Kommunikation digitaler Bilddaten in der Pneumokoniosevorsorge

Pneumokoniose-Vorsorgeuntersuchungen erfordern die Befundung einer Röntgenthoraxaufnahme nach ILO-Staublungenklassifikation. Inzwischen werden die benötigten Aufnahmen bereits in großem Umfang digital angefertigt und kommuniziert. Hierdurch entstehen neue Anforderungen an verwendete Technik und Workflow-Mechanismen, um einen effizienten Ablauf von Untersuchung, Befundung und Dokumentation zu gewährleisten

Similarities and Differences of Mental Health in Women and Men: A Systematic Review of Findings in Three Large German Cohorts

In Germany, large, population-based cohort studies have been implemented in order to identify risk and protective factors for maintaining health across the life span. The purpose of this systematic review is to analyse findings from three large ongoing cohorts and to identify sex-specific prevalence rates, risk and protective factors for mental health. Published studies from the Cooperative Health Research in the Region Augsburg (KORA), the Study of Health in Pomerania (SHIP) and the Gutenberg Health Study (GHS)), representing the southern, north-eastern and middle parts of Germany, were identified through searches of the databases PubMed and Web of Science. A total of 52 articles was identified from the start of each cohort until June 2019. Articles reporting prevalence rates of mental health [N = 22], explanatory factors for mental health [N = 25], or both [N = 5] were identified. Consistent across cohorts, higher prevalence rates of internalizing disorders were found for women and more externalizing disorders for men. Risk and protective factors for mental health included social factors, lifestyle, physical health, body mass index (BMI), diabetes, genetic and biological factors. In all areas, differences and similarities were found between women and men. The most evident were the sex-specific risk profiles for depression with mostly external risk factors for men and internal risk factors for women. Gender was not assessed directly, therefore we examined whether socioeconomic and family-related factors reflecting gender roles or institutionalized gender could be used as a proxy for gender. Overall, this systematic review shows differences and similarities in prevalence rates and determinants of mental health indicators between women and men. They underline the importance of focussing on sex specific approaches in mental health research and in the development of prevention measures. Current research on mental health still lacks focus on gender aspects. Therefore, an increased focus on sex and gender in mental health research is of great importance

Imaging CO₂ reinjection into basalts at the CarbFix2 reinjection reservoir (Hellisheiði, Iceland) with body‐wave seismic interferometry

As part of the Synergetic Utilisation of CO₂ storage Coupled with geothermal EnErgy Deployment project, investigating CO₂ reinjection with different seismic methods, both passive and active seismic surveys have been conducted at the geothermal power plant at Hellisheiði, Iceland. During the 2021 survey, two geophone lines recorded noise for a week. We process the passive-source data with seismic interferometry to image the subsurface structure around the CarbFix2 reinjection reservoir. To improve image quality, we perform an illumination analysis to select only noise panels dominated by body-wave energy. The results show that most noise panels are dominated by air-wave energy arriving from the direction of the power plant. We use panels with a near-vertical incidence to create a zero-offset image and a larger selection of body-wave-dominated panels to create virtual common-shot gathers. We process the gathers with a simple reflection seismology processing workflow to obtain stacked images. The zero-offset images show a relatively lower signal-to-noise ratio and only horizontal reflectors. The stacked images show slightly dipping reflectors and possibly lateral amplitude variations around the expected injection region. This could indicate a region of interest for future research into the reinjection reservoir.ISSN:0016-8025ISSN:1365-247