64 research outputs found

    A streamlined mass spectrometry-based proteomics workflow for large scale FFPE tissue analysis

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    Formalin fixation and paraffin-embedding (FFPE) is the most common method to preserve human tissue for clinical diagnosis, and FFPE archives represent an invaluable resource for biomedical research. Proteins in FFPE material are stable over decades but their efficient extraction and streamlined analysis by mass spectrometry (MS)-based proteomics has so far proven challenging. Herein we describe a MS-based proteomic workflow for quantitative profiling of large FFPE tissue cohorts directly from histopathology glass slides. We demonstrate broad applicability of the workflow to clinical pathology specimens and variable sample amounts, including low-input cancer tissue isolated by laser microdissection. Using state-of-the-art data dependent acquisition (DDA) and data independent acquisition (DIA) MS workflows, we consistently quantify a large part of the proteome in 100 min single-run analyses. In an adenoma cohort comprising more than 100 samples, total workup took less than a day. We observed a moderate trend towards lower protein identification in long-term stored samples (>15 years), but clustering into distinct proteomic subtypes was independent of archival time. Our results underscore the great promise of FFPE tissues for patient phenotyping using unbiased proteomics and they prove the feasibility of analyzing large tissue cohorts in a robust, timely, and streamlined manner

    Control of human adenovirus type 5 gene expression by cellular Daxx/ATRX chromatin-associated complexes

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    Death domain–associated protein (Daxx) cooperates with X-linked α-thalassaemia retardation syndrome protein (ATRX), a putative member of the sucrose non-fermentable 2 family of ATP-dependent chromatin-remodelling proteins, acting as the core ATPase subunit in this complex, whereas Daxx is the targeting factor, leading to histone deacetylase recruitment, H3.3 deposition and transcriptional repression of cellular promoters. Despite recent findings on the fundamental importance of chromatin modification in host-cell gene regulation, it remains unclear whether adenovirus type 5 (Ad5) transcription is regulated by cellular chromatin remodelling to allow efficient virus gene expression. Here, we focus on the repressive role of the Daxx/ATRX complex during Ad5 replication, which depends on intact protein–protein interaction, as negative regulation could be relieved with a Daxx mutant that is unable to interact with ATRX. To ensure efficient viral replication, Ad5 E1B-55K protein inhibits Daxx and targets ATRX for proteasomal degradation in cooperation with early region 4 open reading frame protein 6 and cellular components of a cullin-dependent E3-ubiquitin ligase. Our studies illustrate the importance and diversity of viral factors antagonizing Daxx/ATRX-mediated repression of viral gene expression and shed new light on the modulation of cellular chromatin remodelling factors by Ad5. We show for the first time that cellular Daxx/ATRX chromatin remodelling complexes play essential roles in Ad gene expression and illustrate the importance of early viral proteins to counteract cellular chromatin remodelling

    ARDD 2020: from aging mechanisms to interventions

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    Aging is emerging as a druggable target with growing interest from academia, industry and investors. New technologies such as artificial intelligence and advanced screening techniques, as well as a strong influence from the industry sector may lead to novel discoveries to treat age-related diseases. The present review summarizes presentations from the 7th Annual Aging Research and Drug Discovery (ARDD) meeting, held online on the 1st to 4th of September 2020. The meeting covered topics related to new methodologies to study aging, knowledge about basic mechanisms of longevity, latest interventional strategies to target the aging process as well as discussions about the impact of aging research on society and economy. More than 2000 participants and 65 speakers joined the meeting and we already look forward to an even larger meeting next year. Please mark your calendars for the 8th ARDD meeting that is scheduled for the 31st of August to 3rd of September, 2021, at Columbia University, USA

    The Environmental Problem and Environmental Policy in Economically Open Political One- and Multilevel-Systems and Their Meaning for the EU

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    Die vorliegende Arbeit analysiert im Rahmen allgemeiner Gleichgewichtsmodelle offener Volkswirtschaften mögliche Wechselwirkungen zwischen zunehmender wirtschaftlicher Integration und national bzw. supranational bestimmter Umweltpolitik.Die Arbeit gliedert sich in 3 Teile. Der erste Teil der Arbeit erläutert die internationale Dimension des Umweltproblems, der zweite Teil trägt die modelltheoretischen Bausteine zusammen, mit denen Umweltpolitik in offenen Volkswirtschaftn unter Effizienz-und Verteilungsapekten analysiert werden kann, im dritten Teil dienen diese einer normativen und positiven Analyse der Umweltpolitik in der EU

    Analyzing urban sprawl using multi-temporal and multi-source geospatial data fusion

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    Rapid urban sprawl and unmonitored urbanization of the peri-urban environment are impor-tant challenges for land managers and urban planners in many Asian cities. Several studies show the potential of remote sensing techniques for monitoring the impact of urban expan-sion on physical and ecological sources over time (e.g. Haack et al. 1997, Jensen et al. 1999, Weng 2008). However, many methodological issues remain: one is the aspect of proper measurement and quantification of urban growth, with landscape metrics offering a possible geo-statistical solution. Another issue stems from the fact that convenient remote sensing data is only available for a rather short period of time, while historical land-cover changes derived from analogue maps could further enhance the assessment of urbanization patterns
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