1,720,993 research outputs found
An optimized protocol for the enrichment of small vesicles from murine and non-human primate heart tissue
No full textIn recent years, the interest in extracellular vesicles (EV) functions has been growing. However, methods
for isolating these small vesicles from tissue are still not trivial. Few protocols are available that allows
EV isolation from whole tissue samples, including the heart. Those protocols are based on organ
perfusion using the Langendorff method. In this work, aiming to analysing in vivo biology of small EVs,
we implemented a simple method to obtain enrichment of these vesicles from murine heart tissue. We
tested a titration of Liberase for tissue digestion, which was subjected to differential ultracentrifugation
combined with iodixanol cushion and presented the step-by-step procedure of this protocol. Validation
was done with Nanoparticle Tracking Analysis, transmission Electron Microscope and Western Blot
analysis of EV markers and organelles contaminants. Furthermore, we tested the suitability of the
protocol for isolating EVs from heart tissue obtained from a pre-clinical translational non-human primate
animal model. Therefore, this protocol may be suitable for isolating vesicle from human heart tissue.
Additionally, this method can potentially be applied beyond heart tissue
Human induced pluripotent stem cells for live cell cycle monitoring and endogenous gene activation
No full textOpen-Access-Publikationsfonds 202
Inhibition of Prolyl-Hydroxylase Domain Enzymes Protects From Reoxygenation Injury in Engineered Human Myocardium
No full text
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Physiologists as medical scientists: An early warning from the German academic system
No full textAbstract “Medical scientists” are postgraduate investigators who are engaged in biomedical research, and either hold a biomedical PhD or are qualified in medicine but do not participate in patient care. Medical scientists constitute ~40% of staff at medical faculties and >90% at nonuniversity medical research institutions in Germany. However, medical scientists in Germany face limited long‐term career prospects and a lack of dedicated training and support programmes. They also face time limits on their career progression arising from national academic employment legislation, and imminent reforms by the German government are likely to make this worse. Nevertheless, recent developments in the educational landscape including the introduction of increasingly focused MSc, pre‐PhD, and doctoral programmes to train medically aware basic scientists, as well as improved general recognition of the roles and relevance of medical scientists in health research, are encouraging. Physiologists have taken essential steps to improve the recognition of medical scientists in Germany by introducing a “specialist physiologist” qualification; this initiative could be applied to support medical scientists in other fields and countries. In this review, we describe the particular challenges facing medical scientists in Germany and make recommendations that may apply to other academic systems.Deutsche Forschungsgemeinschaft https://doi.org/10.13039/50110000165
Single-cell transcriptomics reveal extracellular vesicles secretion with a cardiomyocyte proteostasis signature during pathological remodeling
No full textAbstract Aberrant Wnt activation has been reported in failing cardiomyocytes. Here we present single cell transcriptome profiling of hearts with inducible cardiomyocyte-specific Wnt activation (β-cat Δex3 ) as well as with compensatory and failing hypertrophic remodeling. We show that functional enrichment analysis points to an involvement of extracellular vesicles (EVs) related processes in hearts of β-cat Δex3 mice. A proteomic analysis of in vivo cardiac derived EVs from β-cat Δex3 hearts has identified differentially enriched proteins involving 20 S proteasome constitutes, protein quality control (PQC), chaperones and associated cardiac proteins including α-Crystallin B (CRYAB) and sarcomeric components. The hypertrophic model confirms that cardiomyocytes reacted with an acute early transcriptional upregulation of exosome biogenesis processes and chaperones transcripts including CRYAB, which is ameliorated in advanced remodeling. Finally, human induced pluripotent stem cells (iPSC)-derived cardiomyocytes subjected to pharmacological Wnt activation recapitulated the increased expression of exosomal markers, CRYAB accumulation and increased PQC signaling. These findings reveal that secretion of EVs with a proteostasis signature contributes to early patho-physiological adaptation of cardiomyocytes, which may serve as a read-out of disease progression and can be used for monitoring cellular remodeling in vivo with a possible diagnostic and prognostic role in the future.Open-Access-Publikationsfonds 202
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