295 research outputs found
Secretome profiling of heterotypic spheroids suggests a role of fibroblasts in HIF-1 pathway modulation and colorectal cancer photodynamic resistance
No full textPrevious analyses of the tumor microenvironment (TME) have resulted in a concept that tumor progression may depend on interactions between cancer cells and its surrounding stroma. An important aspect of these interactions is the ability of cancer cells to modulate stroma behavior, and vice versa, through the action of a variety of soluble mediators. Here, we aimed to identify soluble factors present in the TME of colorectal cancer cells that may affect relevant pathways through secretome profiling. To partially recapitulate the TME and its architecture, we co-cultured colorectal cancer cells (SW480, TC) with stromal fibroblasts (MRC-5, F) as 3D-spheroids. Subsequent characterization of both homotypic (TC) and heterotypic (TC+F) spheroid secretomes was performed using label-free liquid chromatography-mass spectrometry (LC-MS). Results Through bioinformatic analysis using the NCI-Pathway Interaction Database (NCI-PID) we found that the HIF-1 signaling pathway was most highly enriched among the proteins whose secretion was enhanced in the heterotypic spheroids. Previously, we found that HIF-1 may be associated with resistance of colorectal cancer cells to photodynamic therapy (PDT), an antitumor therapy that combines photosensitizing agents, O2 and light to create a harmful photochemical reaction. Here, we found that the presence of fibroblasts considerably diminished the sensitivity of colorectal cancer cells to photodynamic activity. Although the biological significance of the HIF-1 pathway of secretomes was decreased after photosensitization, this decrease was partially reversed in heterotypic 3D-spheroids. HIF-1 pathway modulation by both PDT and stromal fibroblasts was confirmed through expression assessment of the HIF-target VEGF, as well as through HIF transcriptional activity assessment. Collectively, our results delineate a potential mechanism by which stromal fibroblasts may enhance colorectal cancer cell survival and photodynamic treatment resistance via HIF-1 pathway modulation.Fil: Lamberti, María Julia. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Biotecnologia Ambiental y Salud. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Biotecnologia Ambiental y Salud.; ArgentinaFil: Rettel, Mandy. European Molecular Biology Laboratory; AlemaniaFil: Krijgsveld, Jeroen. German Cancer Research Center; AlemaniaFil: Rivarola, Viviana Alicia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Rumie Vittar, Natalia Belen. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin
Binge drinking
No full textRuntime 1:30 minutesThis resource is provided for informational purposes only and may not reflect current scientific knowledge or medical recommendations.Welcome to Public Health Moment from the University of Minnesota. Binge drinking is common among active-duty military personnel, according to a new study released by the University of Minnesota and the Centers for Disease Control and Prevention (CDC). In the study, involving more than 16,000 military personnel, binge drinking was reported by 43 percent of military personnel during the past month. How does one define binge drinking? Lead author of the study, Mandy Stahre, a University of Minnesota Ph.D. student, explains. Stahre says the study shows that binge drinking is a significant public health problem. With another Public Health Moment, I’m John Finnegan.Finnegan, John; Mandy Stahre. (2009). Binge drinking. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/257616
The Human Leader: Leading from Weakness
No full textHow might human limitations empower rather than impede our leadership? Mandy Smith, author of The Vulnerable Pastor, will explore themes related to vulnerability and weakness in leadership and ministry. Interspersing teaching with guided roundtable discussion, this seminar will empower you to lead out of your deep humanity
Changes in science content knowledge and attitudes toward science teaching of educators attending a zoo-based neuroscience professional development
No full textInformal learning environments often host teachers for learning opportunities, but little is known about the impact of these experiences on teacher professional development (PD). This article describes a unique collaborative PD experience between zoological park personnel and university faculty, examining the impact on teacher content knowledge, attitudes, and classroom lessons. Our findings suggest that the PD improved science content, but made no impact on already high attitudes toward science. In light of the high level of self-reported satisfaction and high frequency of teacher lesson plan use, we propose that the PD had other positive outcomes such as pedagogical knowledge and authentic learning experiences.John L. Pecore, PhD, is an Assistant Professor in the School of Education, College of
Professional Studies at the University of West Florida, Pensacola, FL. Mandy L.
Kirchgessner is a doctoral candidate in Curriculum, Instruction and Technology in
Education at Temple University, Philadelphia, PA. Laura L. Carruth, PhD, is an
Associate Professor at the Neuroscience Institute, Georgia State University, Atlanta, GA.John L. Pecore , Mandy L. Kirchgessner & Laura L. Carruth (2013) Changes in Science Content Knowledge
and Attitudes toward Science Teaching of Educators Attending a Zoo-based Neuroscience Professional Development, The
Clearing House: A Journal of Educational Strategies, Issues and Ideas, 86:6, 238-245Journal Articl
Revealing the Arabidopsis AtGRP7 mRNA binding proteome by specific enhanced RNA interactome capture
No full textReichel M, Schmidt O, Rettel M, et al. Revealing the Arabidopsis AtGRP7 mRNA binding proteome by specific enhanced RNA interactome capture. BMC Plant Biology . 2024;24(1): 552.BACKGROUND: The interaction of proteins with RNA in the cell is crucial to orchestrate all steps of RNA processing. RNA interactome capture (RIC) techniques have been implemented to catalogue RNA- binding proteins in the cell. In RIC, RNA-protein complexes are stabilized by UV crosslinking in vivo. Polyadenylated RNAs and associated proteins are pulled down from cell lysates using oligo(dT) beads and the RNA-binding proteome is identified by quantitative mass spectrometry. However, insights into the RNA-binding proteome of a single RNA that would yield mechanistic information on how RNA expression patterns are orchestrated, are scarce.; RESULTS: Here, we explored RIC in Arabidopsis to identify proteins interacting with a single mRNA, using the circadian clock-regulated Arabidopsis thaliana GLYCINE-RICH RNA-BINDING PROTEIN 7 (AtGRP7) transcript, one of the most abundant transcripts in Arabidopsis, as a showcase. Seedlings were treated with UV light to covalently crosslink RNA and proteins. The AtGRP7 transcript was captured from cell lysates with antisense oligonucleotides directed against the 5'untranslated region (UTR). The efficiency of RNA capture was greatly improved by using locked nucleic acid (LNA)/DNA oligonucleotides, as done in the enhanced RIC protocol. Furthermore, performing a tandem capture with two rounds of pulldown with the 5'UTR oligonucleotide increased the yield. In total, we identified 356 proteins enriched relative to a pulldown from atgrp7 mutant plants. These were benchmarked against proteins pulled down from nuclear lysates by AtGRP7 in vitro transcripts immobilized on beads. Among the proteins validated by in vitro interaction we found the family of Acetylation Lowers Binding Affinity (ALBA) proteins. Interaction of ALBA4 with the AtGRP7 RNA was independently validated via individual-nucleotide resolution crosslinking and immunoprecipitation (iCLIP). The expression of the AtGRP7 transcript in an alba loss-of-function mutant was slightly changed compared to wild-type, demonstrating the functional relevance of the interaction.; CONCLUSION: We adapted specific RNA interactome capture with LNA/DNA oligonucleotides for use in plants using AtGRP7 as a showcase. We anticipate that with further optimization and up scaling the protocol should be applicable for less abundant transcripts. © 2024. The Author(s)
Revealing the Arabidopsis mRNA binding proteome by specific enhanced RNA interactome capture
No full textReichel M, Schmidt O, Rettel M, et al. Revealing the Arabidopsis mRNA binding proteome by specific enhanced RNA interactome capture. bioRxiv. 2024.**Abstract**
**Background**
The interaction of proteins with RNA in the cell is crucial to orchestrate all steps of RNA processing. RNA interactome capture (RIC) techniques have been implemented to catalogue RNA-binding proteins in the cell. In RIC, RNA-protein complexes are stabilized by UV crosslinkingin vivo. Polyadenylated RNAs and associated proteins are pulled down from cell lysates using oligo(dT) beads and the RNA-binding proteome is identified by quantitative mass spectrometry. However, insights into the RNA-binding proteome of a single RNA that would yield mechanistic information on how RNA expression patterns are orchestrated, are scarce.
**Results**
Here, we explored RIC in Arabidopsis to identify proteins interacting with a single mRNA, using the circadian clock-regulatedArabidopsis thalianaGLYCINE-RICH RNA-BINDING PROTEIN 7 (AtGRP7) transcript, one of the most abundant transcripts in Arabidopsis, as a showcase. Seedlings were treated with UV light to covalently crosslink RNA and proteins. TheAtGRP7transcript was captured from cell lysates with antisense oligonucleotides directed against the 5’untranslated region (UTR). The efficiency of RNA capture was greatly enhanced by using locked nucleic acid (LNA)/DNA oligonucleotides, as done in the enhanced RIC protocol. Furthermore, performing a tandem capture with two rounds of pulldown with the 5’UTR oligonucleotide increased the yield. In total, we identified 356 proteins enriched relative to a pulldown fromatgrp7mutant plants. These were benchmarked against proteins pulled down from nuclear lysates byAtGRP7 in vitrotranscripts immobilized on beads. Among the proteins validated byin vitrointeraction we found the family of Acetylation Lowers Binding Affinity (ALBA) proteins. Interaction of ALBA4 with theAtGRP7RNA was independently validated via individual-nucleotide resolution crosslinking and immunoprecipitation (iCLIP). The expression of theAtGRP7transcript in analbaloss-of-function mutant was slightly changed compared to wild-type, demonstrating the functional relevance of the interaction.
**Conclusion**
We adapted specific RNA interactome capture with LNA/DNA oligonucleotides for use in plants usingAtGRP7as a showcase. We anticipate that with further optimization and up-scaling the protocol should be applicable for less abundant transcripts.
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Multi-source monitoring data and numerical analyses for the assessment of settlements affecting built-up areas in variable soil conditions
No full textThis paper presents an integrated analysis based on the use of multi-source wide-area datasets consisting of hydro-mechanical properties of geomaterials, in-situ investigations/measurements (e.g. groundwater levels in wells) and innovative space-borne data (i.e. DInSAR techniques) to support numerical analyses aimed at assessing and predicting the settlements affecting built-up areas in variable soil conditions. To this aim, an expeditious procedure was developed and tested with reference to a district in Rotterdam City (The Netherlands) affected by subsidence phenomena due to the presence of heterogeneous settling strata mainly composed by peat and organic soils. The results obtained allowed investigating the role of predisposing factors of the settlement occurrence and assessing the induced damage on buildings. Considering the widespread diffusion of such geohazards, the followed procedure could help the in-charge authorities to carry out activities at urban scale aimed at identifying the areas most affected by subsidence risk and to select the most suitable and sustainable mitigation strategies.Applied MechanicsGeo-engineerin
On unregulated markets and the freedom of media the transition of the East German press after 1989
No full textThis thesis fills a gap in research, literature and our understanding of transitioning media in post-socialist countries. It tells the fascinating and complicated story of a press moving from state control to a Western free press model. The focus lies on the German Democratic Republic (GDR) between the fall of the Berlin Wall in November 1989 until after German unification in October 1990. It is a story untold in English-language literature, and it is a largely ignored part in contemporary German media history. Being written for an international readership while engaging largely with a unique moment in German and international media history, this thesis bridges a gap between national, continental and academic disciplines. Its primary question is in how far the democratic potential that existed in the moment of revolutionary change in 1989/1990 found its institutional and/or political manifestation in the post-socialist East German press. Contrary to current research, it answers this question by approaching it from the perspective of an expanding Western democratic, and market economic order. It is, thus, not concerned with case studies or one press-related sector but looks at structural change on various levels; its focus lies on the simultaneous battles fought over a free press. Core concern is the intersection between the normative role the press holds in a democratic society and that of a newly developing, or rather established expanding Western market economy.
This thesis analyzes three press-related sectors within a transitioning political setting: first, the opening of the GDR to (and sale of) West German print media; second, the reform and building of distribution infrastructures, and, third, emerging East-West joint ventures and subsequent changing newspaper ownerships. While closely interlinked, reforming distribution became the point of conflict over which issues of a “free press” were being debated. This thesis shows that nothing that happened in the GDR before, during and after the transition happened in isolation nor was it an exclusively East German problem. Instead, East Germany became the battle ground for various interests groups, East and West, but with clear and all-overshadowing interests of West German political and economic groups. These groups generally expanded and continued their long-established interests and disputes onto the newly opened political arena and economic market. Aiming for power at different levels, all had an interest in influencing media and its policies to their own advantage or, by simply circumventing them, created situations on the ground that, once put into place, were hard to change.Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2020-08-01The student, Mandy Troger, accepted the attached license on 2018-07-06 at 20:31.The student, Mandy Troger, submitted this Dissertation for approval on 2018-07-06 at 20:43.This Dissertation was approved for publication on 2018-07-10 at 13:34.DSpace SAF Submission Ingestion Package generated from Vireo submission #12765 on 2018-09-27 at 11:16:44Made available in DSpace on 2018-09-27T16:30:20Z (GMT). No. of bitstreams: 2
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TRRAP is essential for regulating the accumulation of mutant and wild-type p53 in lymphoma
No full textTumors accumulate high levels of mutant p53 (mutp53), which contributes to mutp53 gain-of-function (GOF) properties. The mechanisms that underlie such excessive accumulation are not fully understood. To discover regulators of mutp53 protein accumulation, we performed a large-scale RNA interference (RNAi) screen in a Burkitt's lymphoma (BL) cell line model. We identified TRRAP, a constituent of several histone acetyltransferase (HAT) complexes, as a critical positive regulator of both mutp53 and wild-type p53 (wtp53) levels. TRRAP silencing attenuated p53 accumulation in lymphoma and colon cancer models, while TRRAP overexpression increased mutp53 levels, suggesting a role for TRRAP across cancer entities and p53 mutations. Through CRISPR-Cas9 screening, we identified a 109 amino acid region in the N-terminal HEAT repeat region of TRRAP which was crucial for mutp53 stabilization and cell proliferation. Mass spectrometric analysis of the mutp53 interactome indicated that TRRAP silencing caused degradation of mutp53 via the MDM2-proteasome axis. This suggests that TRRAP is vital for maintaining mutp53 levels by shielding it against the natural p53 degradation machinery. To identify drugs that alleviated p53 accumulation similarly to TRRAP silencing, we performed a small molecule drug screen and found that inhibition of histone deacetylases (HDACs), specifically HDACs1/2/3, decreased p53 levels to a comparable extent. In summary, here we identify TRRAP as a key regulator of p53 levels and link acetylation-modifying complexes to p53 protein stability. Our findings may provide clues for therapeutic targeting of mutp53 in lymphoma and other cancers
Empirical fragility curves for settlement-affected buildings: Analysis of different intensity parameters for seven hundred masonry buildings in The Netherlands
No full textThe analysis and prediction of damage to buildings resting on highly compressible fine-grained ‘‘soft soils” containing (organic) clay
and peat are key issues to be addressed for a proper management of subsidence-affected urban areas. Among the probabilistic approaches suggested in literature, those oriented to the generation of empirical fragility curves are particularly promising provided that a comprehensive dataset for both the subsidence-related intensity (SRI) parameters and the corresponding damage severity to buildings is available. Following this line of thought, in the present paper, a rich sample of more than seven hundred monitored (by remote sensing) and surveyed masonry buildings – mainly resting with their (shallow or piled) foundations on soft soils – is analysed in four urban areas of The Netherlands. Probabilistic functions in the form of fragility curves for building damage are retrieved for three different SRI parameters (i.e., differential settlement, rotation and deflection ratio) derived from the processing of Synthetic Aperture Radar (SAR) images by way of a differential interferometric (DInSAR) technique in combination with the severity levels of the damage recorded from the visual inspection of over 700 masonry buildings. As a novelty with respect to earlier similar studies, the work points out the methodological steps to be followed in order to identify the most appropriate SRI parameter among the selected ones. Thus, the objective of the paper is to improve the existing geotechnical forecasting tools for subsidence-affected urban areas, in order to target areas that require more detailed investigations/analyses and/or to select/prioritize foundation repairing/replacing measures
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