Untersuchungen zur Zellkernarchitektur

Verschiedene Arbeiten der letzten Jahre konnten zeigen, dass sich in vielen, verschiedenen Zelltypen genreiches, transkriptionell aktives und früh replizierendes Chromatin bevorzugt im Inneren der Zellkerne aufhält, während das genarme, transkriptionell inaktive und spät replizierende Chromatin vorrangig an der Zellkernperipherie zu finden ist. Dennoch ist bislang noch nicht wirklich verstanden, welche der Chromatineigenschaften, wie die lokale Gendichte, die Expression oder die Replikationszeit, tatsächlich einen ausschlaggebenden Einfluss auf die räumliche Anordnung im Zellkern haben und welche dieser Eigenschaften nur aufgrund ihrer Korrelation mit diesen „dominanten“ Merkmalen eine spezifische Verteilung im Interphasekern aufweisen. Um dieses Problem zu untersuchen, stellten wir Pools aus BAC Klonen von HSA 11, 12, 18 und 19 für R-und G-Banden-spezifische Regionen, genreiche bzw. genarme Segmente sowie für hoch bzw. niedrig exprimierte Gene zusammen. Mit Hilfe der multicolor 3D-FISH Technik, Bildverarbeitung und computergestützter, quantitativer Auswertungen wurde die Lage dieser BAC Pools im Zellkern sowie ihre Anordnung bezüglich ihrer Chromosomenterritorien analysiert. Sowohl in den humanen Lymphozyten wie in den humanen Fibroblasten fanden wir den R-Banden Pool, den genreichen Pool sowie den Pool, der die hoch exprimierten Gene enthielt, weiter im Zellkerninneren als ihre jeweils korrespondierenden Pools (G-Banden, genarmer Pool, bzw. niedrig exprimierte Gene). Für jeden BAC Pool wurde mittels sorgfältiger Datenbankrecherche die mittlere lokale Gendichte, der mittlere GC Gehalt, die Replikationszeit sowie das mittlere Expressionsniveau bestimmt. Anschließend wurde eine Korrelationsanalyse dieser Parameter mit der berechneten mittleren, relativen Position der Pools im Zellkern durchgeführt. Die höchste Korrelation ergab sich für die Gendichte, während wir zeigen konnten, dass das Expressionsniveau, die Zuordnung zu einer R- oder G.Bande, sowie das Replikationstiming offensichtlich so gut wie keinen Einfluss auf die radiale Anordnung des Chromatins im Zellkern hat. Diese radiale Positionierung der verschiedenen Pools spiegelte sich auch in ihrer Anordnung bezüglich der Chromosomenterritorien wieder. Diese zeigen eine polare Anordnung in Bezug auf den Zellkern: Genreiche Segmente waren zum Mittelpunkt des Zellkerns hin orientiert, während die genarmen Segmente in der Hälfte des CTs zu finden waren, die sich in Richtung der Peripherie erstreckte. Etwas weniger deutlich ausgeprägt wurde diese Anordnung auch für die R-/G-Banden Pools sowie für die von der transkriptionellen Aktivität abhängigen Pools beobachtet. Dies spricht für eine deutliche strukturelle Transformation bei der Umwandlung der Metaphasenchromosomen zu den CTs der Interphase, die Territorien haben eine hohe Plastizität. Wir konnten bestätigen, dass die extrem genreiche und hoch transkriptionell aktive Region 11p15.5 oft weit aus ihrem CT herausragt. Ein ähnliches Verhalten konnte jedoch nicht für die ebenfalls sehr genreichen und transkriptionell aktiven Segmente des Chromosoms 12 beobachtet werden, was gegen die Annahme spricht, das dieses Phänomen des „looping outs“ ein typische Anordnung für Chromatinabschnitte mit solchen extremen Eigenschaften ist. Wir konnten ebenfalls keine Unterschiede für die Verteilung der BAC Pools der Chromosomen 12, 18 und 19 bezüglich der Oberfläche der CTs finden. R- und G-Banden, genreiche und genarme Segmente sowie hoch und niedrig exprimierte Gene scheinen gleichmäßig im gesamten Territorium verteilt zu sein. Die äußere, das CT einschließende Oberfläche scheint entgegen der Erwartung offensichtlich kein wichtiger Reaktionsort für besonders genreiche bzw. hoch exprimierte Sequenzen zu sein

Inter- and intra-specific gene-density-correlated radial chromosome territory arrangements are conserved in Old World monkeys

Recently it has been shown that the gene-density correlated radial distribution of human 18 and 19 homologous chromosome territories (CTs) is conserved in higher primates in spite of chromosomal rearrangements that occurred during evolution. However, these observations were limited to apes and New World monkey species. In order to provide further evidence for the evolutionary conservation of gene-density-correlated CT arrangements, we extended our previous study to Old World monkeys. They comprise the remaining species group to be analyzed in order to obtain a comprehensive overview of the nuclear topology of human 18 and 19 homologous CTs in higher primates. In the present study we investigated four lymphoblastoid cell lines from three species of Old World monkeys by three-dimensional fluorescence in situ hybridization (3D-FISH): two individuals of Japanese macaque ( Macaca fuscata), crab-eating macaque ( Macaca fascicularis), and an interspecies hybrid individual between African green monkey (Cercopithecus aethiops) and Patas monkey ( Erythrocebus patas). Our data demonstrate that gene-poor human 18 homologous CTs are located preferentially close to the nuclear periphery, whereas gene-dense human 19 homologous CTs are oriented towards the nuclear center in all cell lines analyzed. The gene-density-correlated positioning of human 18 and 19 homologous CTs is evolutionarily conserved throughout all major higher primate lineages, despite chromosomal inversions, fusions, fissions or reciprocal translocations that occurred in the course of evolution in these species. This remarkable preservation of a gene-density-correlated chromatin arrangement gives further support for a functionally relevant higher-order chromatin architecture. Copyright (C) 2005 S. Karger AG, Basel

Inheritance of gene density–related higher order chromatin arrangements in normal and tumor cell nuclei

A gene density–related difference in the radial arrangement of chromosome territories (CTs) was previously described for human lymphocyte nuclei with gene-poor CT #18 located toward the nuclear periphery and gene-dense CT #19 in the nuclear interior (Croft, J.A., J.M. Bridger, S. Boyle, P. Perry, P. Teague, and W.A. Bickmore. 1999. J. Cell Biol. 145:1119–1131). Here, we analyzed the radial distribution of chromosome 18 and 19 chromatin in six normal cell types and in eight tumor cell lines, some of them with imbalances and rearrangements of the two chromosomes. Our findings demonstrate that a significant difference in the radial distribution of #18 and #19 chromatin is a common feature of higher order chromatin architecture in both normal and malignant cell types. However, in seven of eight tumor cell lines, the difference was less pronounced compared with normal cell nuclei due to a higher fraction of nuclei showing an inverted CT position, i.e., a CT #18 located more internally than a CT #19. This observation emphasizes a partial loss of radial chromatin order in tumor cell nuclei

Pedestrian Crowd Management Experiments: A Data Guidance Paper

Understanding pedestrian dynamics and the interaction of pedestrians with their environment is crucial to the safe and comfortable design of pedestrian facilities. Experiments offer the opportunity to explore the influence of individual factors. In the context of the project CroMa (Crowd Management in transport infrastructures), experiments were conducted with about 1000 participants to test various physical and social psychological hypotheses focusing on people's behaviour at railway stations and crowd management measures. The following experiments were performed: i) Train Platform Experiment, ii) Crowd Management Experiment, iii) Single-File Experiment, iv) Personal Space Experiment, v) Boarding and Alighting Experiment, vi) Bottleneck Experiment and vii) Tiny Box Experiment. This paper describes the basic planning and implementation steps, outlines all experiments with parameters, geometries, applied sensor technologies and pre- and post-processing steps. All data can be found in the pedestrian dynamics data archive.Comment: 58 pages, 19 figures, under review Collective Dynamic

Implications of site specific response analysis

Definition of design earthquake characteristics, more specifically uniform hazard acceleration response spectrum, on the ground surface is the primary component for performance based design of structures and assessment of seismic vulnerabilities in urban environments. The adopted approach for this purpose requires a probabilistic local seismic hazard assessment, definition of representative site profiles down to the engineering bedrock, and 1D or 2D quivalent or nonlinear, total or effective stress site response analyses depending on the complexity and importance of the structures to be built. Thus, a site-specific response analysis starts with the probabilistic estimation of regional seismicity and earthquake source characteristics, soil stratification, engineering properties of encountered soil layers in the soil profile. The local seismic hazard analysis would yield probabilistic uniform hazard acceleration response spectrum on the bedrock outcrop. Thus, site specific response analyses also need to produce a probabilistic uniform hazard acceleration response spectrum on the ground surface. A general review will be presented based on the previous studies conducted by the author and his co-workers in comparison to major observations and methodologies to demonstrate the implications of site-specific response analysis.WOS:000546471200002Scopus - Affiliation ID: 60105072Conference Proceedings Citation Index- ScienceProceedings PaperHaziran2018YÖK - 2017-1

Inter-hospital transfer for mechanical thrombectomy within the supraregional stroke network NEVAS

BACKGROUND Telemedicine stroke networks are mandatory to provide inter-hospital transfer for mechanical thrombectomy (MT). However, studies on patient selection in daily practice are sparse. METHODS Here, we analyzed consecutive patients from 01/2014 to 12/2018 within the supraregional stroke network \textquotedblNeurovascular Network of Southwest Bavaria\textquotedbl (NEVAS) in terms of diagnoses after consultation, inter-hospital transfer and predictors for performing MT. Degree of disability was rated by the modified Rankin Scale (mRS), good outcome was defined as mRS ≤ 2. Successful reperfusion was assumed when the modified thrombolysis in cerebral infarction (mTICI) was 2b-3. RESULTS Of 5722 telemedicine consultations, in 14.1% inter-hospital transfer was performed, mostly because of large vessel occlusion (LVO) stroke. A total of n = 350 patients with LVO were shipped via NEVAS to our center for MT. While n = 52 recanalized spontaneously, MT-treatment was performed in n = 178 patients. MT-treated patients had more severe strokes according to the median National institute of health stroke scale (NIHSS) (16 vs. 13, p < 0.001), were more often treated with intravenous thrombolysis (64.5% vs. 51.7%, p = 0.026) and arrived significantly earlier in our center (184.5 versus 228.0~min, p < 0.001). Good outcome (27.5% vs. 30.8%, p = 0.35) and mortality (32.6% versus 23.5%, p = 0.79) were comparable in MT-treated versus no-MT-treated patients. In patients with middle cerebral artery occlusion in the M1 segment or carotid artery occlusion good outcome was twice as often in the MT-group (21.8% vs. 12.8%, p = 0.184). Independent predictors for performing MT were higher NIHSS (OR 1.096), higher ASPECTS (OR 1.28), and earlier time window (OR 0.99). CONCLUSION Within a telemedicine network stroke care can successfully be organized as only a minority of patients has to be transferred. Our data provide clinical evidence that all MT-eligible patients should be shipped with the fastest transportation modality as possible