TY - JOUR A1 - Bäumer, Nils A1 - Kartha, Kalathil K. A1 - Kumar Allampally, Naveen A1 - Yagai, Shiki A1 - Albuquerque, Rodrigo Q. A1 - Fernández, Gustavo T1 - Exploiting Coordination Isomerism for Controlled Self-Assembly JF - Angewandte Chemie International Edition N2 - We exploited the inherent geometrical isomerism of a PtII complex as a new tool to control supramolecular assembly processes. UV irradiation and careful selection of solvent, temperature, and concentration leads to tunable coordination isomerism, which in turn allows fully reversible switching between two distinct aggregate species (1D fibers↔2D lamellae) with different photoresponsive behavior. Our findings not only broaden the scope of coordination isomerism, but also open up exciting possibilities for the development of novel stimuli-responsive nanomaterials. KW - coordination isomerism KW - photoresponsive behavior KW - self-assembly KW - supramolecular polymers KW - p-conjugated systems Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221362 VL - 58 ER - TY - JOUR A1 - Sol, Jeroen A. H. P. A1 - Dehm, Volker A1 - Hecht, Reinhard A1 - Würthner, Frank A1 - Schenning, Albertus P. H. J. A1 - Debije, Michael G. T1 - Temperature-Responsive Luminescent Solar Concentrators: Tuning Energy Transfer in a Liquid Crystalline Matrix JF - Angewandte Chemie International Edition N2 - Temperature-responsive luminescent solar concentrators (LSCs) have been fabricated in which the Förster resonance energy transfer (FRET) between a donor–acceptor pair in a liquid crystalline solvent can be tuned. At room temperatures, the perylene bisimide (PBI) acceptor is aggregated and FRET is inactive; while after heating to a temperature above the isotropic phase of the liquid crystal solvent, the acceptor PBI completely dissolves and FRET is activated. This unusual temperature control over FRET was used to design a color-tunable LSC. The device has been shown to be highly stable towards consecutive heating and cooling cycles, making it an appealing device for harvesting otherwise unused solar energy. KW - energy transfer KW - fluorescence KW - liquid crystals KW - luminescent solar concentrators KW - perylene dyes Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238778 VL - 57 ER - TY - JOUR A1 - Counsell, John R. A1 - Karda, Rajvinder A1 - Diaz, Juan Antiano A1 - Carey, Louise A1 - Wiktorowicz, Tatiana A1 - Buckley, Suzanne M. K. A1 - Ameri, Shima A1 - Ng, Joanne A1 - Baruteau, Julien A1 - Almeida, Filipa A1 - de Silva, Rohan A1 - Simone, Roberto A1 - Lugarà, Eleonora A1 - Lignani, Gabriele A1 - Lindemann, Dirk A1 - Rethwilm, Axel A1 - Rahim, Ahad A. A1 - Waddington, Simon N. A1 - Howe, Steven J. T1 - Foamy Virus Vectors Transduce Visceral Organs and Hippocampal Structures following In Vivo Delivery to Neonatal Mice JF - Molecular Therapy: Nucleic Acids N2 - Viral vectors are rapidly being developed for a range of applications in research and gene therapy. Prototype foamy virus (PFV) vectors have been described for gene therapy, although their use has mainly been restricted to ex vivo stem cell modification. Here we report direct in vivo transgene delivery with PFV vectors carrying reporter gene constructs. In our investigations, systemic PFV vector delivery to neonatal mice gave transgene expression in the heart, xiphisternum, liver, pancreas, and gut, whereas intracranial administration produced brain expression until animals were euthanized 49 days post-transduction. Immunostaining and confocal microscopy analysis of injected brains showed that transgene expression was highly localized to hippocampal architecture despite vector delivery being administered to the lateral ventricle. This was compared with intracranial biodistribution of lentiviral vectors and adeno-associated virus vectors, which gave a broad, non-specific spread through the neonatal mouse brain without regional localization, even when administered at lower copy numbers. Our work demonstrates that PFV can be used for neonatal gene delivery with an intracranial expression profile that localizes to hippocampal neurons, potentially because of the mitotic status of the targeted cells, which could be of use for research applications and gene therapy of neurological disorders. KW - foamy virus KW - spumavirus KW - viral vector KW - gene therapy KW - vector tropism KW - bioimaging KW - hippocampus Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223379 VL - 12 ER - TY - JOUR A1 - Griemert, Eva-Verena A1 - Schwarzmaier, Susanne M. A1 - Hummel, Regina A1 - Gölz, Christina A1 - Yang, Dong A1 - Neuhaus, Winfried A1 - Burek, Malgorzata A1 - Förster, Carola Y. A1 - Petkovic, Ivan A1 - Trabold, Raimund A1 - Plesnila, Nikolaus A1 - Engelhard, Kristin A1 - Schäfer, Michael K. A1 - Thal, Serge C. T1 - Plasminogen activator inhibitor-1 augments damage by impairing fibrinolysis after traumatic brain injury JF - Annals of Neurology N2 - Objective Plasminogen activator inhibitor-1 (PAI-1) is the key endogenous inhibitor of fibrinolysis, and enhances clot formation after injury. In traumatic brain injury, dysregulation of fibrinolysis may lead to sustained microthrombosis and accelerated lesion expansion. In the present study, we hypothesized that PAI-1 mediates post-traumatic malfunction of coagulation, with inhibition or genetic depletion of PAI-1 attenuating clot formation and lesion expansion after brain trauma. Methods We evaluated PAI-1 as a possible new target in a mouse controlled cortical impact (CCI) model of traumatic brain injury. We performed the pharmacological inhibition of PAI-1 with PAI-039 and stimulation by tranexamic acid, and we confirmed our results in PAI-1–deficient animals. Results PAI-1 mRNA was time-dependently upregulated, with a 305-fold peak 12 hours after CCI, which effectively counteracted the 2- to 3-fold increase in cerebral tissue-type/urokinase plasminogen activator expression. PAI-039 reduced brain lesion volume by 26% at 24 hours and 43% at 5 days after insult. This treatment also attenuated neuronal apoptosis and improved neurofunctional outcome. Moreover, intravital microscopy demonstrated reduced post-traumatic thrombus formation in the pericontusional cortical microvasculature. In PAI-1–deficient mice, the therapeutic effect of PAI-039 was absent. These mice also displayed 13% reduced brain damage compared with wild type. In contrast, inhibition of fibrinolysis with tranexamic acid increased lesion volume by 25% compared with vehicle. Interpretation This study identifies impaired fibrinolysis as a critical process in post-traumatic secondary brain damage and suggests that PAI-1 may be a central endogenous inhibitor of the fibrinolytic pathway, promoting a procoagulatory state and clot formation in the cerebral microvasculature. Ann Neurol 2019;85:667–680 Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228682 VL - 85 ER - TY - JOUR A1 - Figel, Benedikt A1 - Brinkmann, Leonie A1 - Buff, Christine A1 - Heitmann, Carina Y. A1 - Hofmann, David A1 - Bruchmann, Maximilian A1 - Becker, Michael P. I. A1 - Herrmann, Martin J. A1 - Straube, Thomas T1 - Phasic amygdala and BNST activation during the anticipation of temporally unpredictable social observation in social anxiety disorder patients JF - NeuroImage: Clinical N2 - Anticipation of potentially threatening social situations is a key process in social anxiety disorder (SAD). In other anxiety disorders, recent research of neural correlates of anticipation of temporally unpredictable threat suggests a temporally dissociable involvement of amygdala and bed nucleus of the stria terminalis (BNST) with phasic amygdala responses and sustained BNST activation. However, the temporal profile of amygdala and BNST responses during temporal unpredictability of threat has not been investigated in patients suffering from SAD. We used functional magnetic resonance imaging (fMRI) to investigate neural activation in the central nucleus of the amygdala (CeA) and the BNST during anticipation of temporally unpredictable aversive (video camera observation) relative to neutral (no camera observation) events in SAD patients compared to healthy controls (HC). For the analysis of fMRI data, we applied two regressors (phasic/sustained) within the same model to detect temporally dissociable brain responses. The aversive condition induced increased anxiety in patients compared to HC. SAD patients compared to HC showed increased phasic activation in the CeA and the BNST for anticipation of aversive relative to neutral events. SAD patients as well as HC showed sustained activity alterations in the BNST for aversive relative to neutral anticipation. No differential activity during sustained threat anticipation in SAD patients compared to HC was found. Taken together, our study reveals both CeA and BNST involvement during threat anticipation in SAD patients. The present results point towards potentially SAD-specific threat processing marked by elevated phasic but not sustained CeA and BNST responses when compared to HC. KW - FMRI KW - threat anticipation KW - social anxiety disorder KW - bed nucleus of stria terminalis KW - amygdala Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228071 VL - 22 ER - TY - JOUR A1 - Fazzini, Federica A1 - Lamina, Claudia A1 - Fendt, Liane A1 - Schultheiss, Ulla T. A1 - Kotsis, Fruzsina A1 - Hicks, Andrew A. A1 - Meiselbach, Heike A1 - Weissensteiner, Hansi A1 - Forer, Lukas A1 - Krane, Vera A1 - Eckardt, Kai-Uwe A1 - Köttgen, Anna A1 - Kronenberg, Florian T1 - Mitochondrial DNA copy number is associated with mortality and infections in a large cohort of patients with chronic kidney disease JF - Kidney International N2 - Damage of mitochondrial DNA (mtDNA) with reduction in copy number has been proposed as a biomarker for mitochondrial dysfunction and oxidative stress. Chronic kidney disease (CKD) is associated with increased mortality and risk of cardiovascular disease, but the underlying mechanisms remain incompletely understood. Here we investigated the prognostic role of mtDNA copy number for cause-specific mortality in 4812 patients from the German Chronic Kidney Disease study, an ongoing prospective observational national cohort study of patients with CKD stage G3 and A1-3 or G1-2 with overt proteinuria (A3) at enrollment. MtDNA was quantified in whole blood using a plasmid-normalized PCR-based assay. At baseline, 1235 patients had prevalent cardiovascular disease. These patients had a significantly lower mtDNA copy number than patients without cardiovascular disease (fully-adjusted model: odds ratio 1.03, 95% confidence interval [CI] 1.01-1.05 per 10 mtDNA copies decrease). After four years of follow-up, we observed a significant inverse association between mtDNA copy number and all-cause mortality, adjusted for kidney function and cardiovascular disease risk factors (hazard ratio 1.37, 95% CI 1.09-1.73 for quartile 1 compared to quartiles 2-4). When grouped by causes of death, estimates pointed in the same direction for all causes but in a fully-adjusted model decreased copy numbers were significantly lower only in infection-related death (hazard ratio 1.82, 95% CI 1.08-3.08). A similar association was observed for hospitalizations due to infections in 644 patients (hazard ratio 1.19, 95% CI 1.00-1.42 in the fully-adjusted model). Thus, our data support a role of mitochondrial dysfunction in increased cardiovascular disease and mortality risks as well as susceptibility to infections in patients with CKD. KW - chronic kidney disease KW - infections KW - mitochondrial DNA copy number KW - mortality Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227662 VL - 96 ER - TY - JOUR A1 - Lumma, Anna-Lena A1 - Valk, Sofie L. A1 - Böckler, Anne A1 - Vrtička, Pascal A1 - Singer, Tania T1 - Change in emotional self-concept following socio-cognitive training relates to structural plasticity of the prefrontal cortex JF - Brain and Behavior N2 - Introduction Self-referential processing is a key component of the emotional self-concept. Previous studies have shown that emotional self-referential processing is related to structure and function of cortical midline areas such as medial prefrontal cortex (mPFC), and that it can be altered on a behavioral level by specific mental training practices. However, it remains unknown how behavioral training-related change in emotional self-concept content relates to structural plasticity. Methods To address this issue, we examined the relationship between training-induced change in participant's emotional self-concept measured through emotional word use in the Twenty Statement Test and change in cortical thickness in the context of a large-scale longitudinal mental training study called the ReSource Project. Results Based on prior behavioral findings showing increased emotional word use particularly after socio-cognitive training targeting perspective-taking capacities, this study extended these results by revealing that individual differences in the degree to which participants changed their emotional self-concept after training was positively related to cortical thickness change in right mPFC extending to dorsolateral PFC (dlPFC). Furthermore, increased self-related negative emotional word use after training was positively associated with cortical thickness change in left pars orbitalis and bilateral dlPFC. Conclusions Our findings reveal training-related structural brain change in regions known to be involved in self-referential processing and cognitive control, and could indicate a relationship between restructuring of the emotional self-concept content as well as reappraisal of negative aspects and cortical thickness change. As such, our findings can guide the development of psychological interventions targeted to alter specific facets of the self-concept. KW - cortical thickness KW - emotional word use KW - meditation KW - mental training KW - neuroplasticity KW - self-concept content KW - self-descriptions Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-237395 VL - 8 ER - TY - JOUR A1 - Kiser, Dominik P. A1 - Popp, Sandy A1 - Schmitt-Böhrer, Angelika G. A1 - Strekalova, Tatyana A1 - van den Hove, Daniel L. A1 - Lesch, Klaus-Peter A1 - Rivero, Olga T1 - Early-life stress impairs developmental programming in Cadherin 13 (CDH13)-deficient mice JF - Progress in Neuropsychopharmacology & Biological Psychiatry N2 - Objective Cadherin-13 (CDH13), a member of the calcium-dependent cell adhesion molecule family, has been linked to neurodevelopmental disorders, including autism spectrum (ASD) and attention-deficit/hyperactivity (ADHD) disorders, but also to depression. In the adult brain, CDH13 expression is restricted e.g. to the presynaptic compartment of inhibitory GABAergic synapses in the hippocampus and Cdh13 knockout mice show an increased inhibitory drive onto hippocampal CA1 pyramidal neurons, leading to a shift in excitatory/inhibitory balance. CDH13 is also moderating migration of serotonergic neurons in the dorsal raphe nucleus, establishing projections preferentially to the thalamus and cerebellum during brain development. Furthermore, CDH13 is upregulated by chronic stress as well as in depression, suggesting a role in early-life adaptation to stressful experience. Here, we therefore investigated the interaction between Cdh13 variation and neonatal maternal separation (MS) in mice. Methods Male and female wild-type (Cdh13+/+), heterozygous (Cdh13+/−) and homozygous (Cdh13−/−) knockout mice exposed to MS, or daily handling as control, were subjected to a battery of behavioural tests to assess motor activity, learning and memory as well as anxiety-like behaviour. A transcriptome analysis of the hippocampus was performed in an independent cohort of mice which was exposed to MS or handling, but remained naïve for behavioural testing. Results MS lead to increased anxiety-like behaviour in Cdh13−/− mice compared to the other two MS groups. Cdh13−/− mice showed a context-dependent effect on stress- and anxiety-related behaviour, impaired extinction learning following contextual fear conditioning and decreased impulsivity, as well as a mild decrease in errors in the Barnes maze and reduced risk-taking in the light-dark transition test after MS. We also show sex differences, with increased locomotor activity in female Cdh13−/− mice, but unaltered impulsivity and activity in male Cdh13−/− mice. Transcriptome analysis revealed several pathways associated with cell surface/adhesion molecules to be altered following Cdh13 deficiency, together with an influence on endoplasmic reticulum function. Conclusion MS resulted in increased stress resilience, increased exploration and an overall anxiolytic behavioural phenotype in male Cdh13+/+ and Cdh13+/− mice. Cdh13 deficiency, however, obliterated most of the effects caused by early-life stress, with Cdh13−/− mice exhibiting delayed habituation, no reduction of anxiety-like behaviour and decreased fear extinction. Our behavioural findings indicate a role of CDH13 in the programming of and adaptation to early-life stress. Finally, our transcriptomic data support the view of CDH13 as a neuroprotective factor as well as a mediator in cell-cell interactions, with an impact on synaptic plasticity. KW - Cadherin-13 (CDH13) KW - T-cadherin KW - neurodevelopment KW - autism KW - ADHD KW - depression KW - psychiatric disorders KW - early-life stress KW - mouse KW - RNA sequencing KW - endoplasmic reticulum stress KW - adhesion Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325859 VL - 89 ER - TY - RPRT A1 - Geßner, Daniel T1 - Rethinking renewable energy policies for hydrogen – How the intercept of electricity and hydrogen markets can be addressed N2 - A lot of countries have recently published updated hydrogen strategies, often including more ambitious targets for hydrogen production. In parallel, accompanying ramp-up mechanisms are increasingly coming into focus with the first ones already being released. However, these proposals usually translate mechanisms from renewable energy (RE) policy without considering the specific uncertainties, spillovers, and externalities of integrating hydrogen electrolysis into electricity grids. This article details how different aspects of a policy can address the specific issues, namely funding, risk-mitigation, and the complex relation with electricity markets. It shows that, compared to RE policy, subsidies need to emphasize the input side more strongly as price risks and intermittency from electricity markets are more prominent than from hydrogen markets. Also, it proposes a targeted mechanism to capture the positive externality of mitigating excess electricity in the grid while keeping investment security high. Economic policy should consider such approaches before massively scaling support and avoid the design shortcomings experienced with early RE policy. T3 - Würzburg Economic Papers (W. E. P.) - 111 KW - Wasserstoff KW - Öffentliche Förderung KW - Contract for Difference KW - Elektrizitätsmarkt KW - Erneuerbare Energien KW - Hydrogen Policy KW - Renewable Energy Policy KW - Electricity Markets KW - Support Mechanisms KW - Contracts for Difference Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370973 ER - TY - JOUR A1 - Taubenböck, H. A1 - Weigand, M. A1 - Esch, T. A1 - Staab, J. A1 - Wurm, M. A1 - Mast, J. A1 - Dech, S. T1 - A new ranking of the world's largest cities—Do administrative units obscure morphological realities? JF - Remote Sensing of Environment N2 - With 37 million inhabitants, Tokyo is the world's largest city in UN statistics. With this work we call this ranking into question. Usually, global city rankings are based on nationally collected population figures, which rely on administrative units. Sprawling urban growth, however, leads to morphological city extents that may surpass conventional administrative units. In order to detect spatial discrepancies between the physical and the administrative city, we present a methodology for delimiting Morphological Urban Areas (MUAs). We understand MUAs as a territorially contiguous settlement area that can be distinguished from low-density peripheral and rural hinterlands. We design a settlement index composed of three indicators (settlement area, settlement area proportion and density within the settlements) describing a gradient of built-up density from the urban center to the periphery applying a sectoral monocentric city model. We assume that the urban-rural transition can be defined along this gradient. With it, we re-territorialize the conventional administrative units. Our data basis are recent mapping products derived from multi-sensoral Earth observation (EO) data – namely the Global Urban Footprint (GUF) and the GUF Density (GUF-DenS) – providing globally consistent knowledge about settlement locations and densities. For the re-territorialized MUAs we calculate population numbers using WorldPop data. Overall, we cover the 1692 cities with >300,000 inhabitants on our planet. In our results we compare the consistently re-territorialized MUAs and the administrative units as well as their related population figures. We find the MUA in the Pearl River Delta the largest morphologically contiguous urban agglomeration in the world with a calculated population of 42.6 million. Tokyo, in this new list ranked number 2, loses its top position. In rank-size distributions we present the resulting deviations from previous city rankings. Although many MUAs outperform administrative units by area, we find that, contrary to what we assumed, in most cases MUAs are considerably smaller than administrative units. Only in Europe we find MUAs largely outweighing administrative units in extent. KW - city size KW - urban agglomeration KW - rank-size distribution KW - remote sensing KW - global urban footprint KW - urban morphology Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-240634 VL - 232 ER - TY - JOUR A1 - Germain, Dominique P. A1 - Elliott, Perry M. A1 - Falissard, Bruno A1 - Fomin, Victor V. A1 - Hilz, Max J. A1 - Jovanovic, Ana A1 - Kantola, Ilkka A1 - Linhart, Aleš A1 - Renzo, Mignani A1 - Namdar, Mehdi A1 - Nowak, Albina A1 - Oliveira, João-Paulo A1 - Pieroni, Maurizio A1 - Viana-Baptista, Miguel A1 - Wanner, Christoph A1 - Spada, Marco T1 - The effect of enzyme replacement therapy on clinical outcomes in male patients with Fabry disease: A systematic literature review by a European panel of experts JF - Molecular Genetics and Metabolism Reports N2 - Background Enzyme replacement therapy (ERT) with recombinant human α-galactosidase has been available for the treatment of Fabry disease since 2001 in Europe and 2003 in the USA. Treatment outcomes with ERT are dependent on baseline patient characteristics, and published data are derived from heterogeneous study populations. Methods We conducted a comprehensive systematic literature review of all original articles on ERT in the treatment of Fabry disease published up until January 2017. This article presents the findings in adult male patients. Results Clinical evidence for the efficacy of ERT in adult male patients was available from 166 publications including 36 clinical trial publications. ERT significantly decreases globotriaosylceramide levels in plasma, urine, and in different kidney, heart, and skin cell types, slows the decline in estimated glomerular filtration rate, and reduces/stabilizes left ventricular mass and cardiac wall thickness. ERT also improves nervous system, gastrointestinal, pain, and quality of life outcomes. Conclusions ERT is a disease-specific treatment for patients with Fabry disease that may provide clinical benefits on several outcomes and organ systems. Better outcomes may be observed when treatment is started at an early age prior to the development of organ damage such as chronic kidney disease or cardiac fibrosis. Consolidated evidence suggests a dose effect. Data described in male patients, together with female and paediatric data, informs clinical practice and therapeutic goals for individualized treatment. KW - Fabry disease KW - systematic literature review KW - agalsidase beta KW - agalsidase alfa KW - enzyme replacement therapy KW - adult male patients Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232987 VL - 19 ER - TY - JOUR A1 - Flunkert, Julia A1 - Maierhofer, Anna A1 - Dittrich, Marcus A1 - Müller, Tobias A1 - Horvath, Steve A1 - Nanda, Indrajit A1 - Haaf, Thomas T1 - Genetic and epigenetic changes in clonal descendants of irradiated human fibroblasts JF - Experimental Cell Research N2 - To study delayed genetic and epigenetic radiation effects, which may trigger radiation-induced carcinogenesis, we have established single-cell clones from irradiated and non-irradiated primary human fibroblasts. Stable clones were endowed with the same karyotype in all analyzed metaphases after 20 population doublings (PDs), whereas unstable clones displayed mosaics of normal and abnormal karyotypes. To account for variation in radiation sensitivity, all experiments were performed with two different fibroblast strains. After a single X-ray dose of 2 Gy more than half of the irradiated clones exhibited radiation-induced genome instability (RIGI). Irradiated clones displayed an increased rate of loss of chromosome Y (LOY) and copy number variations (CNVs), compared to controls. CNV breakpoints clustered in specific chromosome regions, in particular 3p14.2 and 7q11.21, coinciding with common fragile sites. CNVs affecting the FHIT gene in FRA3B were observed in independent unstable clones and may drive RIGI. Bisulfite pyrosequencing of control clones and the respective primary culture revealed global hypomethylation of ALU, LINE-1, and alpha-satellite repeats as well as rDNA hypermethylation during in vitro ageing. Irradiated clones showed further reduced ALU and alpha-satellite methylation and increased rDNA methylation, compared to controls. Methylation arrays identified several hundred differentially methylated genes and several enriched pathways associated with in vitro ageing. Methylation changes in 259 genes and the MAP kinase signaling pathway were associated with delayed radiation effects (after 20 PDs). Collectively, our results suggest that both genetic (LOY and CNVs) and epigenetic changes occur in the progeny of exposed cells that were not damaged directly by irradiation, likely contributing to radiation-induced carcinogenesis. We did not observe epigenetic differences between stable and unstable irradiated clones. The fact that the DNA methylation (DNAm) age of clones derived from the same primary culture varied greatly suggests that DNAm age of a single cell (represented by a clone) can be quite different from the DNAm age of a tissue. We propose that DNAm age reflects the emergent property of a large number of individual cells whose respective DNAm ages can be highly variable. KW - copy number variation (CNV) KW - delayed radiation effects KW - DNA methylation (DNAm) age KW - global DNA methylation KW - loss of chromosome Y (LOY); KW - methylation array analysis KW - radiation-induced genome instability (RIGI) Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228177 VL - 370 ER - TY - JOUR A1 - Germain, Dominique P. A1 - Arad, Michael A1 - Burlina, Alessandro A1 - Elliott, Perry M. A1 - Falissard, Bruno A1 - Feldt-Rasmussen, Ulla A1 - Hilz, Max J. A1 - Hughes, Derralynn A. A1 - Ortiz, Alberto A1 - Wanner, Christoph A1 - Weidemann, Frank A1 - Spada, Marco T1 - The effect of enzyme replacement therapy on clinical outcomes in female patients with Fabry disease – A systematic literature review by a European panel of experts JF - Molecular Genetics and Metabolism N2 - Background Heterozygous females with Fabry disease have a wide range of clinical phenotypes depending on the nature of their mutation and their X-chromosome inactivation pattern; it is therefore important to examine outcomes of enzyme replacement therapy (ERT) in the female patient population specifically. This paper presents the findings of a systematic literature review of treatment outcomes with ERT in adult female patients. Methods A comprehensive systematic literature review was conducted through January 2017 to retrieve published papers with original data on ERT in the treatment of Fabry disease. The review included all original articles that presented ERT outcomes data on patients with Fabry disease, irrespective of the study type. Results Clinical evidence for the efficacy of ERT in female patients was available from 67 publications including six clinical trial publications, and indicates significant reductions in plasma and urine globotriaosylceramide (GL-3) accumulation (in female patients with elevated pre-treatment levels) and improvements in cardiac parameters and quality of life (QoL). To date, data are insufficient to conclude on the effects of ERT on the nervous system, gastrointestinal manifestations, and pain in female patients with Fabry disease. Conclusions This review of available literature data demonstrates that ERT in adult female patients with Fabry disease has a beneficial effect on GL-3 levels and cardiac outcomes. The current evidence also suggests that ERT may improve QoL in this patient population, though further studies are needed to examine these results. KW - Fabry disease KW - agalsidase alfa KW - agalsidase beta KW - systematic literature review KW - enzyme replacement therapy KW - adult female patients Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232963 VL - 126 ER - TY - JOUR A1 - Omeñaca, Felix A1 - Vázquez, Liliana A1 - Garcia-Corbeira, Pilar A1 - Mesaros, Narcisa A1 - Hanssens, Linda A1 - Dolhain, Jan A1 - Puente Gómez, Ivonne A1 - Liese, Johannes A1 - Knuf, Markus T1 - Immunization of preterm infants with GSK’s hexavalent combined diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated poliovirus-Haemophilus influenzae type b conjugate vaccine: A review of safety and immunogenicity JF - Vaccine N2 - Background Infants with history of prematurity (<37 weeks gestation) and low birth weight (LBW, <2500 g) are at high risk of infection due to functional immaturity of normal physical and immunological defense mechanisms. Despite current recommendations that infants with history of prematurity/LBW should receive routine immunization according to the same schedule and chronological age as full-term infants, immunization is often delayed. Methods Here we summarize 10 clinical studies and 15 years of post-marketing safety surveillance of GSK’s hexavalent vaccine (DTPa-HBV-IPV/Hib), a combined diphtheria-tetanus-acellular-pertussis-hepatitis-B-inactivated-poliovirus-Haemophilus influenzae-type-b (Hib) conjugate vaccine, when administered alone, or co-administered with pneumococcal conjugate, rotavirus, and meningococcal vaccines and respiratory syncytial virus IgG to infants with history of prematurity/LBW in clinical trials. Results At least 92.5% of infants with history of prematurity/LBW as young as 24 weeks gestation in clinical studies were seropositive to all vaccine antigens after 3-dose primary vaccination with GSK’s hexavalent DTPa-HBV-IPV/Hib vaccine, with robust immune responses to booster vaccination. Seropositivity rates and antibody concentrations to hepatitis B and Hib appeared lower in infants with history of prematurity/LBW than term infants. Between 13–30% of medically stable infants with history of prematurity developed apnea after vaccination with GSK’s hexavalent DTPa-HBV-IPV/Hib vaccine; usually after dose 1. The occurrence of post-immunization cardiorespiratory events appears to be influenced by the severity of any underlying neonatal condition. Most cardiorespiratory events resolve spontaneously or require minimal intervention. GSK’s hexavalent DTPa-HBV-IPV/Hib vaccine was well tolerated in co-administration regimens. Conclusion GSK’s hexavalent DTPa-HBV-IPV/Hib vaccine alone or co-administered with other pediatric vaccines has a clinically acceptable safety and immunogenicity profile when used in infants with history of prematurity/LBW for primary and booster vaccination. Additional studies are needed in very premature and very LBW infants. However, currently available data support using GSK’s hexavalent DTPa-HBV-IPV/Hib vaccine to immunize infants with history of prematurity/LBW according to chronological age. KW - DTPa-HBV-IPV/Hib KW - hexavalent vaccine KW - primary vaccination KW - preterm KW - premature Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234450 VL - 36 ER - TY - JOUR A1 - Spada, Marco A1 - Baron, Ralf A1 - Elliott, Perry M. A1 - Falissard, Bruno A1 - Hilz, Max J. A1 - Monserrat, Lorenzo A1 - Tøndel, Camilla A1 - Tylki-Szymańska, Anna A1 - Wanner, Christoph A1 - Germain, Dominique P. T1 - The effect of enzyme replacement therapy on clinical outcomes in paediatric patients with Fabry disease – A systematic literature review by a European panel of experts JF - Molecular Genetics and Metabolism N2 - Background Fabry disease is caused by a deficiency of the lysosomal enzyme α-galactosidase, resulting in progressive accumulation of globotriaosylceramide (GL-3). The disease can manifest early during childhood and adolescence. Enzyme replacement therapy (ERT) with recombinant human α-galactosidase is the first specific treatment for Fabry disease and has been available in Europe since 2001. This paper presents the findings of a systematic literature review of clinical outcomes with ERT in paediatric patients with Fabry disease. Methods A comprehensive systematic review of published literature on ERT in Fabry disease was conducted in January 2017. The literature analysis included all original articles reporting outcomes of ERT in paediatric patients. Results Treatment-related outcomes in the paediatric population were reported in six publications derived from open-label clinical trials and in 10 publications derived from observational or registry-based studies. ERT was shown to significantly reduce plasma and urine GL-3 levels in paediatric patients with Fabry disease. The effect of ERT on GL-3 clearance from renal podocytes appeared to be agalsidase dose-dependent. ERT relieved pain and improved gastrointestinal symptoms and quality of life. Conclusions Based on the published literature, the use of ERT in paediatric patients can significantly clear GL-3 accumulation, ameliorate the early symptoms of Fabry disease, and improve quality of life. Treatment with ERT in paediatric patients with Fabry disease may be important to prevent further disease progression and overt organ damage. KW - Fabry disease KW - agalsidase alfa KW - agalsidase beta KW - systematic literature review KW - enzyme replacement therapy KW - paediatric patients Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239287 VL - 126 ER - TY - JOUR A1 - Argyrousi, Elentina K. A1 - de Nijs, Laurence A1 - Lagatta, Davi C. A1 - Schlütter, Anna A1 - Weidner, Magdalena T. A1 - Zöller, Johanna A1 - van Goethem, Nick P. A1 - Joca, Sâmia R. L. A1 - van den Hove, Daniel L. A. A1 - Prickaerts, Jos T1 - Effects of DNA methyltransferase inhibition on pattern separation performance in mice JF - Neurobiology of Learning and Memory N2 - Enhancement of synaptic plasticity through changes in neuronal gene expression is a prerequisite for improved cognitive performance. Moreover, several studies have shown that DNA methylation is able to affect the expression of (e.g. plasticity) genes that are important for several cognitive functions. In this study, the effect of the DNA methyltransferase (DNMT) inhibitor RG108 was assessed on object pattern separation (OPS) task in mice. In addition, its effect on the expression of target genes was monitored. Administration of RG108 before the test led to a short-lasting, dose-dependent increase in pattern separation memory that was not present anymore after 48 h. Furthermore, treatment with RG108 did not enhance long-term memory of the animals when tested after a 24 h inter-trial interval in the same task. At the transcriptomic level, acute treatment with RG108 was accompanied by increased expression of Bdnf1, while expression of Bdnf4, Bdnf9, Gria1 and Hdac2 was not altered within 1 h after treatment. Methylation analysis of 14 loci in the promoter region of Bdnf1 revealed a counterintuitive increase in the levels of DNA methylation at three CpG sites. Taken together, these results indicate that acute administration of RG108 has a short-lasting pro-cognitive effect on object pattern separation that could be explained by increased Bdnf1 expression. The observed increase in Bdnf1 methylation suggests a complex interplay between Bdnf methylation-demethylation that promotes Bdnf1 expression and associated cognitive performance. Considering that impaired pattern separation could constitute the underlying problem of a wide range of mental and cognitive disorders, pharmacological agents including DNA methylation inhibitors that improve pattern separation could be compelling targets for the treatment of these disorders. In that respect, future studies are needed in order to determine the effect of chronic administration of such agents. KW - object pattern separation KW - DNA methyltransferase inhibitors KW - BDNF KW - CpG islands KW - epigenetics KW - hippocampal plasticity Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221226 VL - 159 ER - TY - JOUR A1 - Feistauer, Daniela A1 - Richter, Tobias T1 - Validity of students’ evaluations of teaching: Biasing effects of likability and prior subject interest JF - Studies in Educational Evaluation N2 - This study examined the validity of students’ evaluations of teaching as an instrument for measuring teaching quality by examining the effects of likability and prior subject interest as potential biasing effects, measured at the beginning of the course and at the time of evaluation. University students (N = 260) evaluated psychology courses in one semester at a German university with a standardized questionnaire, yielding 517 data points. Cross-classified multilevel analyses revealed fixed effects of likability at both times of measurement and fixed effects of prior subject interest measured at the beginning of the course. Likability seems to exert a substantial bias on student evaluations of teaching, albeit one that is overestimated when measured at the time of evaluation. In contrast, prior subject interest seems to introduce a weak bias. Considering that likability bears no conceptual relationship to teaching quality, these findings point to a compromised validity of students’ evaluations of teaching. KW - cross-classified multilevel analysis KW - likability KW - prior subject interest KW - student evaluations of teaching KW - variance components Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228005 VL - 59 ER - TY - JOUR A1 - Grayston, Rebecca A1 - Czanner, Gabriela A1 - Elhadd, Kareim A1 - Goebel, Andreas A1 - Frank, Bernhard A1 - Üçeyler, Nurcan A1 - Malik, Rayaz A A1 - Alam, Uazman T1 - A systematic review and meta-analysis of the prevalence of small fiber pathology in fibromyalgia: Implications for a new paradigm in fibromyalgia etiopathogenesis JF - Seminars in Arthritis and Rheumatism N2 - Objectives Fibromyalgia is a condition which exhibits chronic widespread pain with neuropathic pain features and has a major impact on health-related quality of life. The pathophysiology remains unclear, however, there is increasing evidence for involvement of the peripheral nervous system with a high prevalence of small fiber pathology (SFP). The aim of this systematic literature review is to establish the prevalence of SFP in fibromyalgia. Methods An electronic literature search was performed using MEDLINE, EMBASE, PubMed, Web of Science, CINAHL and the Cochrane Library databases. Published full-text, English language articles that provide SFP prevalence data in studies of fibromyalgia of patients over 18years old were included. All articles were screened by two independent reviewers using a priori criteria. Methodological quality and risk of bias were evaluated using the critical appraisal tool by Munn et al. Overall and subgroup pooled prevalence were calculated by random-effects meta-analysis with 95% CI. Results Database searches found 935 studies; 45 articles were screened of which 8 full text articles satisfied the inclusion criteria, providing data from 222 participants. The meta-analysis demonstrated the pooled prevalence of SFP in fibromyalgia is 49% (95% CI: 38–60%) with a moderate degree of heterogeneity, (I2= 68%). The prevalence estimate attained by a skin biopsy was 45% (95% CI: 32–59%, I2= 70%) and for corneal confocal microscopy it was 59% (95% CI: 40–78%, I2= 51%). Conclusion There is a high prevalence of SFP in fibromyalgia. This study provides compelling evidence of a distinct phenotype involving SFP in fibromyalgia. Identifying SFP will aid in determining its relationship to pain and potentially facilitate the development of future interventions and pharmacotherapy. KW - small nerve fibres KW - pain KW - fibromyalgia KW - skin biopsy KW - corneal confocal microscopy Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227566 VL - 48 ER - TY - JOUR A1 - Schubert, Frank K. A1 - Hagedorn, Nicolas A1 - Yoshii, Taishi A1 - Helfrich-Förster, Charlotte A1 - Rieger, Dirk T1 - Neuroanatomical details of the lateral neurons of Drosophila melanogaster support their functional role in the circadian system JF - Journal of Comparative Neurology N2 - Drosophila melanogaster is a long-standing model organism in the circadian clock research. A major advantage is the relative small number of about 150 neurons, which built the circadian clock in Drosophila. In our recent work, we focused on the neuroanatomical properties of the lateral neurons of the clock network. By applying the multicolor-labeling technique Flybow we were able to identify the anatomical similarity of the previously described E2 subunit of the evening oscillator of the clock, which is built by the 5th small ventrolateral neuron (5th s-LNv) and one ITP positive dorsolateral neuron (LNd). These two clock neurons share the same spatial and functional properties. We found both neurons innervating the same brain areas with similar pre- and postsynaptic sites in the brain. Here the anatomical findings support their shared function as a main evening oscillator in the clock network like also found in previous studies. A second quite surprising finding addresses the large lateral ventral PDF-neurons (l-LNvs). We could show that the four hardly distinguishable l-LNvs consist of two subgroups with different innervation patterns. While three of the neurons reflect the well-known branching pattern reproduced by PDF immunohistochemistry, one neuron per brain hemisphere has a distinguished innervation profile and is restricted only to the proximal part of the medulla-surface. We named this neuron “extra” l-LNv (l-LNvx). We suggest the anatomical findings reflect different functional properties of the two l-LNv subgroups. KW - circadian clock neurons KW - Drosophila melanogaster KW - flybow KW - morphology KW - RRID: AB_760350 KW - RRID: AB_2315460 KW - RRID: AB_2314242 KW - RRID: AB_2315311 KW - RRID: AB_2314041 KW - RRID: AB_300798 Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234477 VL - 526 ER - TY - JOUR A1 - Chen, Dan A1 - Gehringer, Matthias A1 - Lorenz, Sonja T1 - Developing Small-Molecule Inhibitors of HECT-Type Ubiquitin Ligases for Therapeutic Applications: Challenges and Opportunities JF - ChemBioChem N2 - The ubiquitin system regulates countless physiological and disease-associated processes and has emerged as an attractive entryway for therapeutic efforts. With over 600 members in the human proteome, ubiquitin ligases are the most diverse class of ubiquitylation enzymes and pivotal in encoding specificity in ubiquitin signaling. Although considerable progress has been made in the identification of small molecules targeting RING ligases, relatively little is known about the “druggability” of HECT (homologous to E6AP C terminus) ligases, many of which are critically implicated in human pathologies. A major obstacle to optimizing the few available ligands is our incomplete understanding of their inhibitory mechanisms and the structural basis of catalysis in HECT ligases. Here, we survey recent approaches to manipulate the activities of HECT ligases with small molecules to showcase the particular challenges and opportunities these enzymes hold as therapeutic targets. KW - drug discovery KW - enzymes KW - inhibitors KW - reaction mechanisms KW - structure-activity relationships KW - ubiquitin Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222412 VL - 19 ER - TY - JOUR A1 - Wilson, Duncan A1 - Ambler, Gareth A1 - Lee, Keon-Joo A1 - Lim, Jae-Sung A1 - Shiozawa, Masayuki A1 - Koga, Masatoshi A1 - Li, Linxin A1 - Lovelock, Caroline A1 - Chabriat, Hugues A1 - Hennerici, Michael A1 - Wong, Yuen Kwun A1 - Mak, Henry Ka Fung A1 - Prats-Sánchez, Luis A1 - Martínez-Domeño, Alejandro A1 - Inamura, Shigeru A1 - Yoshifuji, Kazuhisa A1 - Arsava, Ethem Murat A1 - Horstmann, Solveig A1 - Purrucker, Jan A1 - Lam, Bonnie Yin Ka A1 - Wong, Adrian A1 - Kim, Young Dae A1 - Song, Tae-Jin A1 - Schrooten, Maarten A1 - Lemmens, Robin A1 - Eppinger, Sebastian A1 - Gattringer, Thomas A1 - Uysal, Ender A1 - Tanriverdi, Zeynep A1 - Bornstein, Natan M A1 - Ben Assayag, Einor A1 - Hallevi, Hen A1 - Tanaka, Jun A1 - Hara, Hideo A1 - Coutts, Shelagh B A1 - Hert, Lisa A1 - Polymeris, Alexandros A1 - Seiffge, David J A1 - Lyrer, Philippe A1 - Algra, Ale A1 - Kappelle, Jaap A1 - Salman, Rustam Al-Shahi A1 - Jäger, Hans R A1 - Lip, Gregory Y H A1 - Mattle, Heinrich P A1 - Panos, Leonidas D A1 - Mas, Jean-Louis A1 - Legrand, Laurence A1 - Karayiannis, Christopher A1 - Phan, Thanh A1 - Gunkel, Sarah A1 - Christ, Nicolas A1 - Abrigo, Jill A1 - Leung, Thomas A1 - Chu, Winnie A1 - Chappell, Francesca A1 - Makin, Stephen A1 - Hayden, Derek A1 - Williams, David J A1 - Kooi, M Eline A1 - van Dam-Nolen, Dianne H K A1 - Barbato, Carmen A1 - Browning, Simone A1 - Wiegertjes, Kim A1 - Tuladhar, Anil M A1 - Maaijwee, Noortje A1 - Guevarra, Christine A1 - Yatawara, Chathuri A1 - Mendyk, Anne-Marie A1 - Delmaire, Christine A1 - Köhler, Sebastian A1 - van Oostenbrugge, Robert A1 - Zhou, Ying A1 - Xu, Chao A1 - Hilal, Saima A1 - Gyanwali, Bibek A1 - Chen, Christopher A1 - Lou, Min A1 - Staals, Julie A1 - Bordet, Régis A1 - Kandiah, Nagaendran A1 - de Leeuw, Frank-Erik A1 - Simister, Robert A1 - van der Lugt, Aad A1 - Kelly, Peter J A1 - Wardlaw, Joanna M A1 - Soo, Yannie A1 - Fluri, Felix A1 - Srikanth, Velandai A1 - Calvet, David A1 - Jung, Simon A1 - Kwa, Vincent I H A1 - Engelter, Stefan T A1 - Peters, Nils A1 - Smith, Eric E A1 - Yakushiji, Yusuke A1 - Necioglu Orken, Dilek A1 - Fazekas, Franz A1 - Thijs, Vincent A1 - Heo, Ji Hoe A1 - Mok, Vincent A1 - Veltkamp, Roland A1 - Ay, Hakan A1 - Imaizumi, Toshio A1 - Gomez-Anson, Beatriz A1 - Lau, Kui Kai A1 - Jouvent, Eric A1 - Rothwell, Peter M A1 - Toyoda, Kazunori A1 - Bae, Hee-Yoon A1 - Marti-Fabregas, Joan A1 - Werring, David J T1 - Cerebral microbleeds and stroke risk after ischaemic stroke or transient ischaemic attack: a pooled analysis of individual patient data from cohort studies JF - The Lancet Neurology N2 - Background Cerebral microbleeds are a neuroimaging biomarker of stroke risk. A crucial clinical question is whether cerebral microbleeds indicate patients with recent ischaemic stroke or transient ischaemic attack in whom the rate of future intracranial haemorrhage is likely to exceed that of recurrent ischaemic stroke when treated with antithrombotic drugs. We therefore aimed to establish whether a large burden of cerebral microbleeds or particular anatomical patterns of cerebral microbleeds can identify ischaemic stroke or transient ischaemic attack patients at higher absolute risk of intracranial haemorrhage than ischaemic stroke. Methods We did a pooled analysis of individual patient data from cohort studies in adults with recent ischaemic stroke or transient ischaemic attack. Cohorts were eligible for inclusion if they prospectively recruited adult participants with ischaemic stroke or transient ischaemic attack; included at least 50 participants; collected data on stroke events over at least 3 months follow-up; used an appropriate MRI sequence that is sensitive to magnetic susceptibility; and documented the number and anatomical distribution of cerebral microbleeds reliably using consensus criteria and validated scales. Our prespecified primary outcomes were a composite of any symptomatic intracranial haemorrhage or ischaemic stroke, symptomatic intracranial haemorrhage, and symptomatic ischaemic stroke. We registered this study with the PROSPERO international prospective register of systematic reviews, number CRD42016036602. Findings Between Jan 1, 1996, and Dec 1, 2018, we identified 344 studies. After exclusions for ineligibility or declined requests for inclusion, 20 322 patients from 38 cohorts (over 35 225 patient-years of follow-up; median 1·34 years [IQR 0·19–2·44]) were included in our analyses. The adjusted hazard ratio [aHR] comparing patients with cerebral microbleeds to those without was 1·35 (95% CI 1·20–1·50) for the composite outcome of intracranial haemorrhage and ischaemic stroke; 2·45 (1·82–3·29) for intracranial haemorrhage and 1·23 (1·08–1·40) for ischaemic stroke. The aHR increased with increasing cerebral microbleed burden for intracranial haemorrhage but this effect was less marked for ischaemic stroke (for five or more cerebral microbleeds, aHR 4·55 [95% CI 3·08–6·72] for intracranial haemorrhage vs 1·47 [1·19–1·80] for ischaemic stroke; for ten or more cerebral microbleeds, aHR 5·52 [3·36–9·05] vs 1·43 [1·07–1·91]; and for ≥20 cerebral microbleeds, aHR 8·61 [4·69–15·81] vs 1·86 [1·23–2·82]). However, irrespective of cerebral microbleed anatomical distribution or burden, the rate of ischaemic stroke exceeded that of intracranial haemorrhage (for ten or more cerebral microbleeds, 64 ischaemic strokes [95% CI 48–84] per 1000 patient-years vs 27 intracranial haemorrhages [17–41] per 1000 patient-years; and for ≥20 cerebral microbleeds, 73 ischaemic strokes [46–108] per 1000 patient-years vs 39 intracranial haemorrhages [21–67] per 1000 patient-years). Interpretation In patients with recent ischaemic stroke or transient ischaemic attack, cerebral microbleeds are associated with a greater relative hazard (aHR) for subsequent intracranial haemorrhage than for ischaemic stroke, but the absolute risk of ischaemic stroke is higher than that of intracranial haemorrhage, regardless of cerebral microbleed presence, antomical distribution, or burden. Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233710 VL - 18 ER - TY - JOUR A1 - Waszak, Sebastian M A1 - Northcott, Paul A A1 - Buchhalter, Ivo A1 - Robinson, Giles W A1 - Sutter, Christian A1 - Groebner, Susanne A1 - Grund, Kerstin B A1 - Brugières, Laurence A1 - Jones, David T W A1 - Pajtler, Kristian W A1 - Morrissy, A Sorana A1 - Kool, Marcel A1 - Sturm, Dominik A1 - Chavez, Lukas A1 - Ernst, Aurelie A1 - Brabetz, Sebastian A1 - Hain, Michael A1 - Zichner, Thomas A1 - Segura-Wang, Maia A1 - Weischenfeldt, Joachim A1 - Rausch, Tobias A1 - Mardin, Balca R A1 - Zhou, Xin A1 - Baciu, Cristina A1 - Lawerenz, Christian A1 - Chan, Jennifer A A1 - Varlet, Pascale A1 - Guerrini-Rousseau, Lea A1 - Fults, Daniel W A1 - Grajkowska, Wiesława A1 - Hauser, Peter A1 - Jabado, Nada A1 - Ra, Young-Shin A1 - Zitterbart, Karel A1 - Shringarpure, Suyash S A1 - De La Vega, Francisco M A1 - Bustamante, Carlos D A1 - Ng, Ho-Keung A1 - Perry, Arie A1 - MacDonald, Tobey J A1 - Driever, Pablo Hernáiz A1 - Bendel, Anne E A1 - Bowers, Daniel C A1 - McCowage, Geoffrey A1 - Chintagumpala, Murali M A1 - Cohn, Richard A1 - Hassall, Timothy A1 - Fleischhack, Gudrun A1 - Eggen, Tone A1 - Wesenberg, Finn A1 - Feychting, Maria A1 - Lannering, Birgitta A1 - Schüz, Joachim A1 - Johansen, Christoffer A1 - Andersen, Tina V A1 - Röösli, Martin A1 - Kuehni, Claudia E A1 - Grotzer, Michael A1 - Kjaerheim, Kristina A1 - Monoranu, Camelia M A1 - Archer, Tenley C A1 - Duke, Elizabeth A1 - Pomeroy, Scott L A1 - Shelagh, Redmond A1 - Frank, Stephan A1 - Sumerauer, David A1 - Scheurlen, Wolfram A1 - Ryzhova, Marina V A1 - Milde, Till A1 - Kratz, Christian P A1 - Samuel, David A1 - Zhang, Jinghui A1 - Solomon, David A A1 - Marra, Marco A1 - Eils, Roland A1 - Bartram, Claus R A1 - von Hoff, Katja A1 - Rutkowksi, Stefan A1 - Ramaswamy, Vijay A1 - Gilbertson, Richard J A1 - Korshunov, Andrey A1 - Taylor, Michael D A1 - Lichter, Peter A1 - Malkin, David A1 - Gajjar, Amar A1 - Korbel, Jan O A1 - Pfister, Stefan M T1 - Spectrum and prevalence of genetic predisposition in medulloblastoma: a retrospective genetic study and prospective validation in a clinical trial cohort JF - The Lancet Oncology N2 - Background Medulloblastoma is associated with rare hereditary cancer predisposition syndromes; however, consensus medulloblastoma predisposition genes have not been defined and screening guidelines for genetic counselling and testing for paediatric patients are not available. We aimed to assess and define these genes to provide evidence for future screening guidelines. Methods In this international, multicentre study, we analysed patients with medulloblastoma from retrospective cohorts (International Cancer Genome Consortium [ICGC] PedBrain, Medulloblastoma Advanced Genomics International Consortium [MAGIC], and the CEFALO series) and from prospective cohorts from four clinical studies (SJMB03, SJMB12, SJYC07, and I-HIT-MED). Whole-genome sequences and exome sequences from blood and tumour samples were analysed for rare damaging germline mutations in cancer predisposition genes. DNA methylation profiling was done to determine consensus molecular subgroups: WNT (MBWNT), SHH (MBSHH), group 3 (MBGroup3), and group 4 (MBGroup4). Medulloblastoma predisposition genes were predicted on the basis of rare variant burden tests against controls without a cancer diagnosis from the Exome Aggregation Consortium (ExAC). Previously defined somatic mutational signatures were used to further classify medulloblastoma genomes into two groups, a clock-like group (signatures 1 and 5) and a homologous recombination repair deficiency-like group (signatures 3 and 8), and chromothripsis was investigated using previously established criteria. Progression-free survival and overall survival were modelled for patients with a genetic predisposition to medulloblastoma. Findings We included a total of 1022 patients with medulloblastoma from the retrospective cohorts (n=673) and the four prospective studies (n=349), from whom blood samples (n=1022) and tumour samples (n=800) were analysed for germline mutations in 110 cancer predisposition genes. In our rare variant burden analysis, we compared these against 53 105 sequenced controls from ExAC and identified APC, BRCA2, PALB2, PTCH1, SUFU, and TP53 as consensus medulloblastoma predisposition genes according to our rare variant burden analysis and estimated that germline mutations accounted for 6% of medulloblastoma diagnoses in the retrospective cohort. The prevalence of genetic predispositions differed between molecular subgroups in the retrospective cohort and was highest for patients in the MBSHH subgroup (20% in the retrospective cohort). These estimates were replicated in the prospective clinical cohort (germline mutations accounted for 5% of medulloblastoma diagnoses, with the highest prevalence [14%] in the MBSHH subgroup). Patients with germline APC mutations developed MBWNT and accounted for most (five [71%] of seven) cases of MBWNT that had no somatic CTNNB1 exon 3 mutations. Patients with germline mutations in SUFU and PTCH1 mostly developed infant MBSHH. Germline TP53 mutations presented only in childhood patients in the MBSHH subgroup and explained more than half (eight [57%] of 14) of all chromothripsis events in this subgroup. Germline mutations in PALB2 and BRCA2 were observed across the MBSHH, MBGroup3, and MBGroup4 molecular subgroups and were associated with mutational signatures typical of homologous recombination repair deficiency. In patients with a genetic predisposition to medulloblastoma, 5-year progression-free survival was 52% (95% CI 40–69) and 5-year overall survival was 65% (95% CI 52–81); these survival estimates differed significantly across patients with germline mutations in different medulloblastoma predisposition genes. Interpretation Genetic counselling and testing should be used as a standard-of-care procedure in patients with MBWNT and MBSHH because these patients have the highest prevalence of damaging germline mutations in known cancer predisposition genes. We propose criteria for routine genetic screening for patients with medulloblastoma based on clinical and molecular tumour characteristics. Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233425 VL - 19 ER - TY - JOUR A1 - Schwarz, Christopher A1 - Scharf, Lennart T. A1 - Scherpf, Thorsten A1 - Weismann, Julia A1 - Gessner, Viktoria H. T1 - Isolation of the Metalated Ylides [Ph3P−C−CN]M (M=Li, Na, K): Influence of the Metal Ion on the Structure and Bonding Situation JF - Chemistry – A European Journal N2 - The isolation and structural characterization of the cyanido-substituted metalated ylides [Ph3P−C−CN]M (1-M; M=Li, Na, K) are reported with lithium, sodium, and potassium as metal cations. In the solid-state, most different aggregates could be determined depending on the metal and additional Lewis bases. The crown-ether complexes of sodium (1-Na) and potassium (1-K) exhibited different structures, with sodium preferring coordination to the nitrogen end, whereas potassium binds in an unusual η2-coordination mode to the two central carbon atoms. The formation of the yldiide was accompanied by structural changes leading to shorter C−C and longer C−N bonds. This could be attributed to the delocalization of the free electron pairs at the carbon atom into the antibonding orbitals of the CN moiety, which was confirmed by IR spectroscopy and computational studies. Detailed density functional theory calculations show that the changes in the structure and the bonding situation were most pronounced in the lithium compounds due to the higher covalency. KW - alkali metals KW - bond theory KW - lithium KW - structure elucidation KW - solid-state structures KW - ylides Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-235445 VL - 25 ER - TY - JOUR A1 - Faber, T. A1 - Hudec, M. A1 - Malinský, M. A1 - Meinzinger, P. A1 - Porod, W. A1 - Staub, F. T1 - A unified leptoquark model confronted with lepton non-universality in B-meson decays JF - Physics Letters B N2 - The anomalies in the B-meson sector, in particular R-K(*) and R-D(*), are often interpreted as hints for physics beyond the Standard Model. To this end, leptoquarks or a heavy Z' represent the most popular SM extensions which can explain the observations. However, adding these fields by hand is not very satisfactory as it does not address the big questions like a possible embedding into a unified gauge theory. On the other hand, light leptoquarks within a unified framework are challenging due to additional constraints such as lepton flavor violation. The existing accounts typically deal with this issue by providing estimates on the relevant couplings. In this letter we consider a complete model based on the SU(4)(C) circle times SU(2)(L) circle times U(1) R gauge symmetry, a subgroup of SO(10), featuring both scalar and vector leptoquarks. We demonstrate that this setup has, in principle, all the potential to accommodate R-K(*) and R-D(*) while respecting bounds from other sectors usually checked in this context. However, it turns out that K-L -> e(+/-)mu(-/+) severely constraints not only the vector but also the scalar leptoquarks and, consequently, also the room for any sizeable deviations of R-K(*) from 1. We briefly comment on the options for extending the model in order to conform this constraint. Moreover, we present a simple criterion for all-orders proton stability within this class of models. Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227419 VL - 787 ER - TY - JOUR A1 - Müntze, Jonas A1 - Gensler, Daniel A1 - Maniuc, Octavian A1 - Liu, Dan A1 - Cairns, Tereza A1 - Oder, Daniel A1 - Hu, Kai A1 - Lorenz, Kristina A1 - Frantz, Stefan A1 - Wanner, Christoph A1 - Nordbeck, Peter T1 - Oral Chaperone Therapy Migalastat for Treating Fabry Disease: Enzymatic Response and Serum Biomarker Changes After 1 Year JF - Clinical Pharmacology & Therapeutics N2 - Long-term effects of migalastat therapy in clinical practice are currently unknown. We evaluated migalastat efficacy and biomarker changes in a prospective, single-center study on 14 patients with Fabry disease (55 ± 14 years; 11 men). After 1 year of open-label migalastat therapy, patients showed significant changes in alpha-galactosidase-A activity (0.06–0.2 nmol/minute/mg protein; P = 0.001), left ventricular myocardial mass index (137–130 g/m2; P = 0.037), and serum creatinine (0.94–1.0 mg/dL; P = 0.021), accounting for deterioration in estimated glomerular filtration rate (87–78 mL/minute/1.73 m2; P = 0.012). The enzymatic increase correlated with myocardial mass reduction (r = −0.546; P = 0.044) but not with renal function (r = −0.086; P = 0.770). Plasma globotriaosylsphingosine was reduced in therapy-naive patients (10.9–6.0 ng/mL; P = 0.021) and stable (9.6–12.1 ng/mL; P = 0.607) in patients switched from prior enzyme-replacement therapy. These first real-world data show that migalastat substantially increases alpha-galactosidase-A activity, stabilizes related serum biomarkers, and improves cardiac integrity in male and female patients with amenable Fabry disease mutations. Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231626 VL - 105 ER - TY - JOUR A1 - Garain, Swadhin A1 - Shoyama, Kazutaka A1 - Ginder, Lea-Marleen A1 - Sárosi, Menyhárt A1 - Würthner, Frank T1 - The delayed box: biphenyl bisimide cyclophane, a supramolecular nano-environment for the efficient generation of delayed fluorescence JF - Journal of the American Chemical Society N2 - Activating delayed fluorescence emission in a dilute solution via a non-covalent approach is a formidable challenge. In this report, we propose a strategy for efficient delayed fluorescence generation in dilute solution using a non-covalent approach via supramolecularly engineered cyclophane-based nanoenvironments that provide sufficient binding strength to π-conjugated guests and that can stabilize triplet excitons by reducing vibrational dissipation and lowering the singlet–triplet energy gap for efficient delayed fluorescence emission. Toward this goal, a novel biphenyl bisimide-derived cyclophane is introduced as an electron-deficient and efficient triplet-generating host. Upon encapsulation of various carbazole-derived guests inside the nanocavity of this cyclophane, emissive charge transfer (CT) states close to the triplet energy level of the biphenyl bisimide are generated. The experimental results of host–guest studies manifest high association constants up to 10\(^4\) M\(^{–1}\) as the prerequisite for inclusion complex formation, the generation of emissive CT states, and triplet-state stabilization in a diluted solution state. By means of different carbazole guest molecules, we could realize tunable delayed fluorescence emission in this carbazole-encapsulated biphenyl bisimide cyclophane in methylcyclohexane/carbon tetrachloride solutions with a quantum yield (QY) of up to 15.6%. Crystal structure analyses and solid-state photophysical studies validate the conclusions from our solution studies and provide insights into the delayed fluorescence emission mechanism. KW - aromatic compounds KW - complexation KW - encapsulation KW - fluorescence KW - hydrocarbons Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370385 SN - 0002-7863 VL - 146 IS - 31 ER - TY - THES A1 - Dusel, Marco T1 - Exziton-Polariton-Kondensation in organischen Halbleiter-Mikrokavitäten mit hemisphärischen Potentiallandschaften T1 - Exciton-polariton condensation in organic semiconductor microcavities with hemispherical potential landscapes N2 - Exziton-Polaritonen sind hybride Quasiteilchen, die entstehen durch die starke Kopplung zwischen Halbleiter-Exzitonen und Mikrokavitätsphotonen in einem optischen Resonator. Aufgrund ihres bosonischen Charakters können die Polaritonen Kondensate ausbilden. In dieser Arbeit ist der emittierende organische Halbleiter das fluoreszierende Protein mCherry. Um einen räumlichen Einschluss zu generieren wurden hemisphärische Potentiale genutzt. Durch die Variation der Potentiallandschaft (Linse, Molekül, Kette, Su-Schrieffer-Heeger-Kette und Honigwaben-Gitter) konnten Eigenschaften wie beispielsweise topologisch nicht-triviale Defekte experimentell bei Umgebungstemperatur demonstriert werden. Zusammengefasst beschäftigt sich diese Arbeit mit der Exziton-Polartion Kondensation in unterschiedlichen Potentiallandschaften mit dem organischen Halbleiter mCherry. N2 - Exciton polaritons are hybrid quasiparticles that are created by the strong coupling between semiconductor excitons and microcavity photons in an optical cavity. Due to their bosonic character, the polaritons can form condensates. In this work, the emitting organic semiconductor is the fluorescent protein mCherry. Hemispherical potentials were used to create a spatial confinement. By varying the potential landscape (lens, molecule, chain, Su-Schrieffer-Heeger chain and honeycomb lattice), properties such as topological non-trivial defects were experimentally demonstrated at ambient temperature. In conclusion, this work deals with exciton-polarisation condensation in different potential landscapes with the organic semiconductor mCherry. KW - Exziton-Polariton KW - Kondensation KW - Organischer Halbleiter KW - Optischer Resonator KW - mCherry Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370554 ER - TY - THES A1 - Drakopoulos, Antonios T1 - Opioid receptor oligomerization study through fluorescent selective ligands T1 - Untersuchung der Opioid Rezeptor Oligomerisierung mittels fluoreszierender selektiver Liganden N2 - Opioid receptors (ORs) are among the most intensively studied members of the G protein-coupled receptor (GPCR) family due to their important role in pain management and their involvement in psychological and neurological disorders. However, currently available opioid drugs exhibit both serious drawbacks, such as addiction, and life-threatening side effects, such as respiratory depression. Contrary to the classic monomeric model, indirect evidence suggests that ORs might form dimers, which could be endowed with a distinct pharmacological profile, and, thus, be exploited to develop innovative drugs. However, direct evidence for the spontaneous formation of OR dimers in living cells under physiological condition are missing. The focus of this thesis was the design, synthesis and characterization of new, highly subtype-selective OR fluorescent ligands to be used as tools for state-of-the-art microscopy methods, such as single molecule microscopy (SMM), in heterologous cells and potentially in native tissue, in order to investigate OR organization and mobility on the surface of intact, living cells, at low/physiological expression levels. The μOR is the OR subtype which plays the most critical role in pain modulation, while mediating the effects of the most powerful analgesic drugs. Also, it is the OR subtype which is mostly responsible for the major adverse effects of the currently marketed opioid drugs. We aimed to develop a new μOR-selective fluorescent ligand with a potential irreversible binding mode. Although the approach was in principle successful, i.e. the labelled cells were visible and distinguishable; this initial attempt was not suitable for SMM due to the ligands’ poor selectivity and affinity as well as due to its high background noise. A second generation of the fluorescent ligand was designed; however the synthesis and characterization are part of another doctoral thesis. Lately, δOR has received attention as a promising drug target, due to its distinct pharmacological profile which features low abuse liability and lack of physical dependence. In addition, δOR expression has been associated with cancer regulation in the periphery, thus further highlighting the interest of imaging tools for this receptor. In this thesis, the development and characterization of two new δOR-selective fluorescent probes with excellent optical properties, based on the well-studied ligand naltrindole (NTI) is presented. Their application in SMM studies is currently underway at the group of Prof. Dr. Davide Calebiro at the University of Birmingham. The κOR is a subtype which has also emerged as a drug target due to its low abuse potential. Despite a growing interest in this receptor, κOR-selective fluorescent probes have been particularly scarce in literature. Herein, the design, synthesis and characterization of the first reported set of fluorescent κOR-selective probes with antagonistic properties, based on the established ligand 5’-guanidinonaltrindole (5’-GNTI) is presented. Two of these were employed for SMM experiments to investigate κOR homodimerization, localization and trafficking. Our findings do not support homodimerization of the κOR-bound probe complexes, while showing that the majority of them follow a normal Brownian diffusion on the cell surface. N2 - Opioid-Rezeptoren (OR) gehören aufgrund ihrer wesentlichen Rolle bei der Schmerztherapie und ihrer Beteiligung an physiologischen und neurologischen Störungen zu den am intensivsten untersuchten Mitgliedern der G-Protein-gekoppelten Rezeptor (GPCR) Familie. Jedoch haben aktuell erhältliche Opioid-Arzneimittel schwerwiegende Nachteile, wie Abhängigkeit, und lebensbedrohliche Nebenwirkungen, wie Atemdepression. Im Gegensatz zu dem klassischen Monomer-Modell legen indirekte Hinweise nahe, dass ORs Dimere formen können, welche mit einem spezifischen pharmakologischen Profil ausgestattet sein könnten und daher für die Entwicklung innovativer Arzneimittel verwendet werden könnten. Jedoch gibt es keinen direkten Beweis für die spontane Bildung von OR-Dimeren in lebenden Zellen unter physiologischen Bedingungen. Der Fokus dieser Doktorarbeit war daher das Design, die Synthese und Charakterisierung von neuen hoch subtyp-selektiven fluoreszierenden OR Liganden, welche als Hilfsmittel für hochmoderne Mikroskopie-Anwendungen Anwendung finden sollen, wie Einzelmolekül-Mikroskopie (EMM) in heterologen Zellen und potentiell in nativem Gewebe, um OR-Organisierung und Mobilität auf der Oberfläche von intakten lebenden Zellen bei niedrigen/physiologischen Expressions-Spiegeln zu untersuchen. Der μOR ist der OR Subtyp, der die entscheidenste Rolle bei der Schmerzmodulierung spielt, indem er die Wirkung der stärksten analgetischen Arzneien vermittelt. Des Weiteren ist dieser OR-Subtyp der Subtyp, der größtenteils für die wesentlichen unerwünschten Nebenwirkungen der aktuell vermarkteten Opioid-Arzneimittel verantwortlich ist. Das Ziel dieser Arbeit war daher, einen neuen μOR-selektiven fluoreszierenden Liganden mit einem potentiell irreversiblen Bindungsmodus zu entwickeln. Obwohl dieser Ansatz prinzipiell erfolgreich war, das heißt die markierten Zellen waren sicht- und unterscheidbar, war dieser erste Ansatz aufgrund der geringen Selektivität und Affinität des Liganden und aufgrund seines hohen Hintergrundrauschens nicht für EMM geeignet. Daher wurde eine zweite Generation fluoreszierender Liganden entworfen. Deren Synthese und Charakterisierung ist jedoch Teil einer anderen Doktorarbeit. Kürzlich erhielt der δOR aufgrund seines spezifischen pharmakologischen Profils, welches ein geringes Missbrauchsrisiko und das Fehlen körperlicher Abhängigkeit beinhaltet, vielseitige Beachtung als ein vielversprechendes Arznei-Target. Des Weiteren wurde δOR-Expression mit Krebsregulation in der Peripherie assoziiert, was das Interesse an einem bildgebenden Werkzeug für diesen Rezeptor zusätzlich unterstreicht. In dieser Doktorarbeit wird die Entwicklung und Charakterisierung von zwei neuen, auf dem gut untersuchten Liganden Naltrindol (NTI) basierenden, δOR-selektiven fluoreszierenden Sonden mit sehr guten optischen Eigenschaften gezeigt. Ihre Anwendung in EMM Untersuchungen läuft derzeit bei Kooperationspartnern im Arbeitskreis von Professor Davide Calebiro an der Universität Birmingham an. Der κOR ist der Subtyp, der auch als Arznei-Target aufgrund seines geringen Missbrauchspotentials in Erscheinung getreten ist. Obwohl steigendes Interesse an diesem Rezeptor besteht, sind κOR-selektive fluoreszierende Sonden in der Literatur bisher kaum beschrieben. In dieser Arbeit wird das Design, die Synthese und Charakterisierung des ersten beschriebenen Sets von fluoreszierenden κOR-selektiven Sonden mit antagonistischen Eigenschaften, basierend auf dem Liganden 5’-Guanidinonaltrindol (5’-GNTI) gezeigt. Zwei dieser Liganden wurden für EMM Experimente verwendet, um die κOR Homodimerisierung, Lokalisation und Transportwege zu untersuchen. Unsere Ergebnisse zeigen keine Homodimerisierung des κOR-gebundenen Sondenkomplexes und außerdem, dass die Mehrheit der Rezeptoren einer normalen Brown’schen Diffusion auf der Zelloberfläche folgt. KW - Opioidrezeptor KW - fluorescent ligands KW - opioid receptors KW - TIRF microscopy KW - GPCR oligomerization KW - Oligomerisation KW - Ligand KW - Fluoreszierende Liganden KW - GPCR Oligomerisierung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-207179 ER - TY - JOUR A1 - Barkhuizen, Melinda A1 - van Mechelen, Ralph A1 - Vermeer, Marijne A1 - Chedraui, Peter A1 - Paes, Dean A1 - van den Hove, Daniel L. A. A1 - Vaes, Bart A1 - Mays, Robert W. A1 - Steinbusch, Harry W. M. A1 - Robertson, Nicola J. A1 - Kramer, Boris W. A1 - Gavilanes, Antonio W. D. T1 - Systemic multipotent adult progenitor cells improve long-term neurodevelopmental outcomes after preterm hypoxic-ischemic encephalopathy JF - Behavioural Brain Research N2 - There is an urgent need for therapies that could reduce the disease burden of preterm hypoxic-ischemic encephalopathy. Here, we evaluate the long-term effects of multipotent adult progenitor cells (MAPC) on long-term behavioral outcomes in a preterm rat model of perinatal asphyxia. Rats of both sexes were treated with two doses of MAPCs within 24 h after the insult. Locomotor, cognitive and psychiatric impairments were evaluated starting at 1.5 (juvenile) and 6 months (adult). Hypoxia-ischemia affected locomotion, cognition, and anxiety in a sex-dependent manner, with higher vulnerability observed in males. The MAPC therapy partially attenuated deficits in object recognition memory in females of all tested ages, and in the adult males. The hypoxic insult caused delayed hyperactivity in adult males, which was corrected by MAPC therapy. These results suggest that MAPCs may have long-term benefits for neurodevelopmental outcome after preterm birth and global hypoxia-ischemia, which warrants further preclinical exploration. KW - hypoxic-ischemic encephalopathy KW - preterm brain KW - stem cell therapy KW - neurodevelopment Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221506 VL - 362 ER - TY - THES A1 - Korte, Pamela T1 - Die funktionelle Bedeutung des Lipidstoffwechsels für die Stomataöffnung bei Hitzestress in \(Arabidopsis\) \(thaliana\) T1 - The functional significance of lipid metabolism for stomatal opening during heat stress in \(Arabidopsis\) \(thaliana\) N2 - Pflanzen sind verschiedenen Umweltbedingungen ausgesetzt, die zu suboptimalen Wachstumsbedingungen führen können. Dies gilt für eine Vielzahl von biotischen und abiotischen Faktoren. In der hier vorgelegten Arbeit wird der Effekt von erhöhten Temperaturen und Hitze genauer analysiert. Hitze ist einer der wichtigsten abiotischen Stressfaktoren, der das Pflanzenwachstum und die Reproduktion beeinflusst. Viele wichtige Kulturpflanzen zeigen immense Ertragseinbußen, die durch Hitze hervorgerufen werden. Durch den fortschreitenden Klimawandel werden jedoch Hitzeperioden immer häufiger und somit die Folgen für die Nahrungsproduktion immer gravierender. Zur Züchtung von Pflanzen die hitzetolerant sind und weniger hohe Ertragseinbußen unter diesem Stress aufweisen, ist es essenziell die grundlegenden molekularen Mechanismen der Hitzetoleranz zu verstehen. Es müssen die verschiedenen physiologischen und biochemischen Prozesse identifiziert werden, die es Pflanzen ermöglichen, sich anzupassen. Es ist bekannt, dass die Anpassungsmechanismen von Pflanzen komplex sind und sowohl Veränderungen auf zellulärer wie auch auf organismischer Ebene beinhalten. Ziel dieser Arbeit war es, weitere Erkenntnisse zu gewinnen, wie diese Anpassung vonstattengeht und welche molekularen Prozesse an ihr beteiligt sind. Ein Hauptaugenmerk lag dabei auf dem Einfluss des Lipidmetabolismus und den daran beteiligten Enzymen. Es konnte bereits gezeigt werden, dass die Akkumulation von Triacylglycerolen bei hohen Temperaturen die basale Thermotoleranz bei Arabidopsis thaliana erhöht. Wie jedoch der genaue Mechanismus dieser durch Triacylglycerole vermittelten Thermotoleranz funktioniert, war bis dato nicht bekannt. Ich konnte zeigen, dass die angesammelten Triacylglycerole genutzt werden können, um die Stomata während des Hitzestress zu öffnen. Dies führt zu einer erhöhten Transpiration und somit einer Kühlung der Blätter. Der Abbau von Triacylglycerolen und Stärke am Morgen ist notwendig, um die Stomata zu öffnen. Zusätzlich dient der Abbau der Aufrechterhaltung des Citratzyklus und somit der Energieversorgung. In weiteren Experimenten konnte ich durch Fütterung mit stabil markierter Laurinsäure zeigen, dass die Triacylglycerole auch dem Aufbau neuer Aminosäuren unter Stressbedingungen dienen. Die hier vorgestellten Arbeiten bieten die Grundlage, um den Mechanismus der Thermotoleranz besser zu verstehen. Das Verständnis der in dieser Arbeit beschriebenen molekularen Signalwege und Enzyme kann langfristig dazu beitragen hitzeresistentere Nutzpflanzen zu züchten. N2 - Plants are exposed to various environmental conditions that can lead to suboptimal growth conditions. This applies to a variety of biotic and abiotic factors. In the work presented here, the effect of elevated temperatures and heat is analyzed in more detail. Heat is one of the most important abiotic stress factors affecting plant growth and reproduction. Many important crops show immense yield losses caused by heat. However, as climate change progresses, periods of heat are becoming more frequent and the consequences for food production are becoming increasingly serious. Understanding the basic molecular mechanisms of heat tolerance is essential to breed plants that are heat tolerant and show less yield loss under this stress. The various physiological and biochemical processes that enable plants to adapt need to be identified. It is known that the adaptation mechanisms of plants are complex and involve changes at both the cellular and organismal level. The aim of this work was to gain further insights into how this adaptation takes place and which molecular processes are involved. The main focus was on the influence of lipid metabolism and the enzymes involved. It has already been shown that the accumulation of triacylglycerols at high temperatures increases basal thermotolerance in Arabidopsis thaliana. However, the exact mechanism of this triacylglycerol mediated thermotolerance was not known until now. I was able to show that the accumulated triacylglycerols can be used to open the stomata during heat stress. This leads to increased transpiration and thus cooling of the leaves. The degradation of triacylglycerols and starch in the morning is necessary to open the stomata. In addition, the degradation serves to maintain the citrate cycle and thus the energy supply. In further experiments, I was able to show by feeding stably labeled lauric acid that the triacylglycerols also serve to build up new amino acids under stress conditions. The work presented here provides the basis for a better understanding of the mechanism of thermotolerance. Understanding the molecular signaling pathways and enzymes described in this work could - in the long term - contribute to breeding of more heat-resistant crops. KW - Hitzestress KW - Ackerschmalwand KW - Arabidopsis thaliana KW - Triacylglycerol Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370461 ER - TY - THES A1 - Brohm, Katharina Andrea T1 - (Differential-) Diagnostik bei primärem Hyperaldosteronismus: Ermittlung eines LC-MS/MS-spezifischen Aldosterongrenzwerts für den Kochsalzbelastungstest und Evaluation des Orthostasetests hinsichtlich der Differenzierung von Subgruppen T1 - (Differential) Diagnosis in Primary Aldosteronism: Determination of an LC-MS/MS-Specific Aldosterone Cut-Off Value for the Saline Infusion Test and Evaluation of the Postural Stimulation Test Regarding the Differentiation of Subtypes N2 - Der primäre Hyperaldosteronismus (PA) stellt aktuell den häufigsten Grund für das Vorliegen einer sekundären Hypertonie dar. Der in der Bestätigungsdiagnostik verwendete Kochsalzbelastungstest basiert dabei auf einem fehlenden Absinken der Aldosteronkonzentration im Testverlauf bei Patient:innen mit PA im Vergleich zu Patient:innen mit essentieller Hypertonie (EH). Die Konzentrationsbestimmung erfolgte bisher mittels Immunoassay. Mit der LC-MS/MS steht jedoch mittlerweile eine weitere wichtige analytische Methode in der quantitativen Bestimmung von Steroidhormonen zur Verfügung, welche in dieser Arbeit im Hinblick auf den Kochsalzbelastungstest untersucht wurde. Hohe Bedeutung kommt außerdem der Subtypdifferenzierung des PA zu, da die Ätiologie der Erkrankung wegweisend für die Art der Therapie ist. Das Ziel dieser Studie war einerseits die Ermittlung eines LC-MS/MS-spezifischen Aldosteron-Cut-off-Wertes im Kochsalzbelastungstest und die Evaluation des Nutzens der Bestimmung von Steroidprofilen in der Diagnostik des PA. Zum anderen wurde der diagnostische Nutzen des Orthostasetests zur Unterscheidung von unilateraler und bilateraler Genese bei vorliegendem PA untersucht. Im Rahmen dieser Studien wurden 187 bzw. 158 Patient:innen analysiert, die zwischen 2009 und 2019 bei Verdacht auf oder Vorliegen eines PA im Universitätsklinikum Würzburg vorstellig wurden. Die Diagnose wurde gemäß der aktuellen Leitlinie anhand der Ergebnisse des Kochsalzbelastungstests, NNVKs, Bildgebung und postoperativen Outcomes gestellt. Mithilfe der LC-MS/MS wurden erneut die Aldosteronkonzentrationen der aufbewahrten Serumproben des Kochsalzbelastungstests, sowie ein erweitertes Steroidpanel bestimmt. Unter Verwendung einer ROC-Analyse wurden die jeweils bestehenden Cut-off-Werte optimiert bzw. neu ermittelt. Die mittels Immunoassay bestimmten Aldosteronkonzentrationen lagen um 28 ng/L höher als die mittels LC-MS/MS bestimmten Konzentrationen. Trotzdem lag der neu ermittelte LC-MS/MS-spezifische Aldosteron-Cut-off-Wert für den Kochsalzbelastungstest bei 69 ng/L und damit höher als der für den Immunoassay geltende, optimierte Aldosteron-Cut-off von 54 ng/L. Unter Verwendung des LC-MS/MS- spezifischen Cut-off-Werts erreichte der Kochsalzbelastungstest eine Sensitivität von 78,6% bei einer Spezifität von 89,3%. Die Sensitivität des Immunoassay-spezifischen Cut-off-Werts betrug 95,2% bei einer Spezifität von 86,9%. Das Bestimmen des gesamten Steroidprofils führte zu keiner zusätzlichen diagnostischen Information bei Durchführung des Kochsalzbelastungstests. Bei Betrachtung der gesamten Patient:innenkohorte erreichte der Orthostasetest, basierend auf einem Absinken der Plasmaaldosteronkonzentration nach 4h in Orthostase um ≥ 28% eine Sensitivität von 36,7% bei einer Spezifität von 100%. Wurde das Vorliegen eines gültigen Tests (Cortisolabfall nach 4h ≥ 10%) oder das Vorliegen einer unilateralen Raumforderung in der Bildgebung vorausgesetzt, stieg die Sensitivität des Orthostasetests auf 51,4% bzw. 51,6% bei gleichbleibend hoher Spezifität von 100% an. Abschließend lässt sich sagen, dass der Orthostasetest keine Alternative zum NNVK darstellt, jedoch als einfache, nicht invasive Methode der zusätzlichen Orientierung zur Untersuchung der Ätiologie des PAs dienen kann. Eine prospektive Evaluation der jeweils neu ermittelten Cut-off-Werte wird notwendig sein, um deren Anwendbarkeit im klinischen Alltag zu überprüfen. Außerdem könnte die Bestimmung der Hybridsteroide 18-Oxocortisol und 18-Hydroxycortisol wegweisend für die Genese des PA sein. N2 - Primary aldosteronism (PA) is currently the most common cause of secondary hypertension. The saline infusion test used in confirmatory diagnostics is based on the lack of decrease in aldosterone concentration during the test in patients with PA compared to those with essential hypertension (EH). Until now, concentration determination has been performed using immunoassay. However, LC-MS/MS has now become an important analytical method for the quantitative determination of steroid hormones, which was investigated in this work in relation to the saline infusion test. Subtype differentiation of PA is also of great significance, as the subtype determines the therapy. The aim of this study was to determine an LC-MS/MS-specific aldosterone cut-off value in the saline infusion test and to evaluate the benefit of determining steroid profiles in the diagnosis of PA. Additionally, the diagnostic value of the postural stimulation test to differentiate between unilateral and bilateral disease in the presence of PA was investigated. In these studies, 187 and 158 patients, respectively, who presented with suspected or confirmed PA at the University Hospital Würzburg between 2009 and 2019 were analyzed. The diagnosis was made according to current guidelines based on the results of the saline infusion test, adrenal vein sampling, imaging, and postoperative outcomes. Using LC-MS/MS, aldosterone concentrations of the stored serum samples from the saline infusion test and an extended steroid panel were determined. ROC analysis was used to optimize or newly determine the existing cut-off values. Aldosterone concentrations determined by immunoassay were 28 ng/L higher than those determined by LC-MS/MS. Nevertheless, the newly determined LC-MS/MS-specific aldosterone cut-off value for the saline infusion test was 69 ng/L, which is higher than the optimized aldosterone cut-off of 54 ng/L for the immunoassay. Using the LC-MS/MS-specific cut-off value, the saline infusion test achieved a sensitivity of 78.6% with a specificity of 89.3%. The sensitivity of the immunoassay-specific cut-off value was 95.2% with a specificity of 86.9%. Determining the entire steroid profile did not provide any additional diagnostic information when performing the saline infusion test. Considering the entire patient cohort, the postural stimulation test, based on a decrease in plasma aldosterone concentration after 4 hours in an upright position by ≥ 28%, achieved a sensitivity of 36.7% with a specificity of 100%. When the test was considered valid (cortisol decrease after 4 hours ≥ 10%) or the presence of a unilateral mass on imaging was assumed, the sensitivity of the postural stimulation test increased to 51.4% and 51.6%, respectively, with a consistently high specificity of 100%. In conclusion, the postural stimulation test does not serve as an alternative to adrenal vein sampling but can provide additional information in investigating the subtype of PA as a simple, non-invasive method. A prospective evaluation of the newly determined cut-off values will be necessary to verify their applicability in clinical practice. Additionally, determining the hybrid steroids 18-oxocortisol and 18-hydroxycortisol could be crucial for understanding the subtype of PA. KW - Aldosteronismus KW - Aldosteron KW - primärer Hyperaldosteronismus KW - LC-MS/MS KW - Kochsalzbelastungstest KW - Orthostasetest Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369382 ER - TY - JOUR A1 - Czimmerer, Zsolt A1 - Daniel, Bence A1 - Horvath, Attila A1 - Rückerl, Dominik A1 - Nagy, Gergely A1 - Kiss, Mate A1 - Peloquin, Matthew A1 - Budai, Marietta M. A1 - Cuaranta-Monroy, Ixchelt A1 - Simandi, Zoltan A1 - Steiner, Laszlo A1 - Nagy Jr., Bela A1 - Poliska, Szilard A1 - Banko, Csaba A1 - Bacso, Zsolt A1 - Schulman, Ira G. A1 - Sauer, Sascha A1 - Deleuze, Jean-Francois A1 - Allen, Judith E. A1 - Benko, Szilvia A1 - Nagy, Laszlo T1 - The Transcription Factor STAT6 Mediates Direct Repression of Inflammatory Enhancers and Limits Activation of Alternatively Polarized Macrophages JF - Immunity N2 - The molecular basis of signal-dependent transcriptional activation has been extensively studied in macrophage polarization, but our understanding remains limited regarding the molecular determinants of repression. Here we show that IL-4-activated STAT6 transcription factor is required for the direct transcriptional repression of a large number of genes during in vitro and in vivo alternative macrophage polarization. Repression results in decreased lineage-determining transcription factor, p300, and RNA polymerase II binding followed by reduced enhancer RNA expression, H3K27 acetylation, and chromatin accessibility. The repressor function of STAT6 is HDAC3 dependent on a subset of IL-4-repressed genes. In addition, STAT6-repressed enhancers show extensive overlap with the NF-κB p65 cistrome and exhibit decreased responsiveness to lipopolysaccharide after IL-4 stimulus on a subset of genes. As a consequence, macrophages exhibit diminished inflammasome activation, decreased IL-1β production, and pyroptosis. Thus, the IL-4-STAT6 signaling pathway establishes an alternative polarization-specific epigenenomic signature resulting in dampened macrophage responsiveness to inflammatory stimuli. KW - IL-4 KW - STAT6 KW - alternative macrophage polarization KW - transcription KW - repression KW - inflammation KW - inflammasome activation KW - pyroptosis KW - IL-1β KW - macrophage epigenomics Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223380 VL - 48 ER - TY - JOUR A1 - Trübe, Patricia A1 - Hertlein, Tobias A1 - Mrochen, Daniel M. A1 - Schulz, Daniel A1 - Jorde, Ilka A1 - Krause, Bettina A1 - Zeun, Julia A1 - Fischer, Stefan A1 - Wolf, Silver A. A1 - Walther, Birgit A1 - Semmler, Torsten A1 - Bröker, Barbara M. A1 - Ulrich, Rainer G. A1 - Ohlsen, Knut A1 - Holtfreter, Silva T1 - Bringing together what belongs together: Optimizing murine infection models by using mouse-adapted Staphylococcus aureus strains JF - International Journal of Medical Microbiology N2 - Staphylococcus (S.) aureus is a leading cause of bacterial infection world-wide, and currently no vaccine is available for humans. Vaccine development relies heavily on clinically relevant infection models. However, the suitability of mice for S. aureus infection models has often been questioned, because experimental infection of mice with human-adapted S. aureus requires very high infection doses. Moreover, mice were not considered to be natural hosts of S. aureus. The latter has been disproven by our recent findings, showing that both laboratory mice, as well as wild small mammals including mice, voles, and shrews, are naturally colonized with S. aureus. Here, we investigated whether mouse-and vole-derived S. aureus strains show an enhanced virulence in mice as compared to the human-adapted strain Newman. Using a step-wise approach based on the bacterial genotype and in vitro assays for host adaptation, we selected the most promising candidates for murine infection models out of a total of 254 S. aureus isolates from laboratory mice as well as wild rodents and shrews. Four strains representing the clonal complexes (CC) 8, 49, and 88 (n = 2) were selected and compared to the human-adapted S. aureus strain Newman (CC8) in murine pneumonia and bacteremia models. Notably, a bank vole-derived CC49 strain, named DIP, was highly virulent in BALB/c mice in pneumonia and bacteremia models, whereas the other murine and vole strains showed virulence similar to or lower than that of Newman. At one tenth of the standard infection dose DIP induced disease severity, bacterial load and host cytokine and chemokine responses in the murine bacteremia model similar to that of Newman. In the pneumonia model, DIP was also more virulent than Newman but the effect was less pronounced. Whole genome sequencing data analysis identified a pore-forming toxin gene, lukF-PV(P83)/lukM, in DIP but not in the other tested S. aureus isolates. To conclude, the mouse-adapted S. aureus strain DIP allows a significant reduction of the inoculation dose in mice and is hence a promising tool to develop clinically more relevant infection models. KW - Staphylococcus aureus KW - host-adapted KW - infection model KW - mouse KW - vole KW - CC49 Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229081 VL - 309 ER - TY - JOUR A1 - Tappenbeck, Nils A1 - Schröder, Hannes M. A1 - Niebergall-Roth, Elke A1 - Hassinger, Fathema A1 - Dehio, Ulf A1 - Dieter, Kathrin A1 - Kraft, Korinna A1 - Kerstan, Andreas A1 - Esterlechner, Jasmina A1 - Frank, Natasha Y. A1 - Scharffetter-Kochanek, Karin A1 - Murphy, George F. A1 - Orgill, Dennis P. A1 - Beck, Joachim A1 - Frank, Markus H. A1 - Ganss, Christoph A1 - Kluth, Mark A. T1 - In vivo safety profile and biodistribution of GMP-manufactured human skin-derived ABCB5-positive mesenchymal stromal cells for use in clinical trials JF - Cytotherapy N2 - Background aims Human dermal ABCB5-expressing mesenchymal stromal cells (ABCB5+ MSCs) represent a promising candidate for stem cell–based therapy of various currently uncurable diseases in several fields of regenerative medicine. We have developed and validated a method to isolate, from human skin samples, and expand ABCB5+ MSCs that meet the guideline criteria of the International Society for Cellular Therapy. We are able to process these cells into a Good Manufacturing Practice–conforming, MSC-based advanced-therapy medicinal product. Methods To support the development of ABCB5+ MSCs for potential therapeutic topical, intramuscular and intravenous administration, we have tested our product in a series of Good Laboratory Practice–compliant nonclinical in-vivo studies addressing all relevant aspects of biosafety, including potential long-term persistence and proliferation, distribution to nontarget tissues, differentiation into undesired cell types, ectopic tissue formation, tumor formation and local tissue reaction. Results (i) Subcutaneous application of 1 × 107 ABCB5+ MSCs/animal and intravenous application of 2 × 106 ABCB5+ MSCs/animal, respectively, to immunocompromised mice did not result in safety-relevant biodistribution, persistence or proliferation of the cells; (ii) three monthly subcutaneous injections of ABCB5+ MSCs at doses ranging from 1 × 105 to 1 × 107 cells/animal and three biweekly intravenous injections of 2 × 106 ABCB5+ MSCs/animal, respectively, to immunocompromised mice were nontoxic and revealed no tumorigenic potential; and (iii) intramuscular injection of 5 × 106 ABCB5+ MSCs/animal to immunocompromised mice was locally well tolerated. Discussion The present preclinical in vivo data demonstrate the local and systemic safety and tolerability of a novel advanced-therapy medicinal product based on human skin-derived ABCB5+ MSCs. KW - stromal cells KW - stem cells KW - MSC KW - biodistribution KW - safety KW - ABCB5 KW - GMP KW - tumorigenicity KW - toxicity KW - persistence Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-240456 VL - 21 ER - TY - JOUR A1 - Göttlich, Claudia A1 - Kunz, Meik A1 - Zapp, Cornelia A1 - Nietzer, Sarah L. A1 - Walles, Heike A1 - Dandekar, Thomas A1 - Dandekar, Gudrun T1 - A combined tissue-engineered/in silico signature tool patient stratification in lung cancer JF - Molecular Oncology N2 - Patient-tailored therapy based on tumor drivers is promising for lung cancer treatment. For this, we combined in vitro tissue models with in silico analyses. Using individual cell lines with specific mutations, we demonstrate a generic and rapid stratification pipeline for targeted tumor therapy. We improve in vitro models of tissue conditions by a biological matrix-based three-dimensional (3D) tissue culture that allows in vitro drug testing: It correctly shows a strong drug response upon gefitinib (Gef) treatment in a cell line harboring an EGFR-activating mutation (HCC827), but no clear drug response upon treatment with the HSP90 inhibitor 17AAG in two cell lines with KRAS mutations (H441, A549). In contrast, 2D testing implies wrongly KRAS as a biomarker for HSP90 inhibitor treatment, although this fails in clinical studies. Signaling analysis by phospho-arrays showed similar effects of EGFR inhibition by Gef in HCC827 cells, under both 2D and 3D conditions. Western blot analysis confirmed that for 3D conditions, HSP90 inhibitor treatment implies different p53 regulation and decreased MET inhibition in HCC827 and H441 cells. Using in vitro data (western, phospho-kinase array, proliferation, and apoptosis), we generated cell line-specific in silico topologies and condition-specific (2D, 3D) simulations of signaling correctly mirroring in vitro treatment responses. Networks predict drug targets considering key interactions and individual cell line mutations using the Human Protein Reference Database and the COSMIC database. A signature of potential biomarkers and matching drugs improve stratification and treatment in KRAS-mutated tumors. In silico screening and dynamic simulation of drug actions resulted in individual therapeutic suggestions, that is, targeting HIF1A in H441 and LKB1 in A549 cells. In conclusion, our in vitro tumor tissue model combined with an in silico tool improves drug effect prediction and patient stratification. Our tool is used in our comprehensive cancer center and is made now publicly available for targeted therapy decisions. KW - 3D lung tumor model KW - Boolean signaling network KW - chemoresistance KW - HSP90 inhibitor KW - insilico drug screening too KW - KRAS mutation signature Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233137 VL - 12 ER - TY - JOUR A1 - Stromecki, Margaret A1 - Tatari, Nazanin A1 - Coudière Morrison, Ludivine A1 - Kaur, Ravinder A1 - Zagozewski, Jamie A1 - Palidwor, Gareth A1 - Ramaswamy, Vijay A1 - Skowron, Patryk A1 - Wölfl, Matthias A1 - Milde, Till A1 - Del Bigio, Marc R. A1 - Taylor, Michael D. A1 - Werbowetski-Ogilvie, Tamra E. T1 - Characterization of a novel OTX2-driven stem cell program in Group 3 and Group 4 medulloblastoma JF - Molecular Oncology N2 - Medulloblastoma (MB) is the most common malignant primary pediatric brain cancer. Among the most aggressive subtypes, Group 3 and Group 4 originate from stem/progenitor cells, frequently metastasize, and often display the worst prognosis, yet we know the least about the molecular mechanisms driving their progression. Here, we show that the transcription factor orthodenticle homeobox 2 (OTX2) promotes self-renewal while inhibiting differentiation in vitro and increases tumor initiation from MB stem/progenitor cells in vivo. To determine how OTX2 contributes to these processes, we employed complementary bioinformatic approaches to characterize the OTX2 regulatory network and identified novel relationships between OTX2 and genes associated with neuronal differentiation and axon guidance signaling in Group 3 and Group 4 MB stem/progenitor cells. In particular, OTX2 levels were negatively correlated with semaphorin (SEMA) signaling, as expression of 9 SEMA pathway genes is upregulated following OTX2 knockdown with some being potential direct OTX2 targets. Importantly, this negative correlation was also observed in patient samples, with lower expression of SEMA4D associated with poor outcome specifically in Group 4 tumors. Functional proof-of-principle studies demonstrated that increased levels of select SEMA pathway genes are associated with decreased self-renewal and growth in vitro and in vivo and that RHO signaling, known to mediate the effects of SEMA genes, is contributing to the OTX2 KD phenotype. Our study provides mechanistic insight into the networks controlled by OTX2 in MB stem/progenitor cells and reveals novel roles for axon guidance genes and their downstream effectors as putative tumor suppressors in MB. KW - axon guidance genes KW - medulloblastoma KW - orthodenticle homeobox 2 KW - RHO KW - semaphorin KW - stem cells Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-240089 VL - 12 ER - TY - JOUR A1 - Hoenigl, Martin A1 - Orasch, Thomas A1 - Faserl, Klaus A1 - Prattes, Juergen A1 - Loeffler, Juergen A1 - Springer, Jan A1 - Gsaller, Fabio A1 - Reischies, Frederike A1 - Duettmann, Wiebke A1 - Raggam, Reinhard B. A1 - Lindner, Herbert A1 - Haas, Hubertus T1 - Triacetylfusarinine C: A urine biomarker for diagnosis of invasive aspergillosis JF - Journal of Infection N2 - Objectives Early diagnosis of invasive aspergillosis (IA) remains challenging, with available diagnostics being limited by inadequate sensitivities and specificities. Triacetylfusarinine C, a fungal siderophore that has been shown to accumulate in urine in animal models, is a potential new biomarker for diagnosis of IA. Methods We developed a method allowing absolute and matrix-independent mass spectrometric quantification of TAFC. Urine TAFC, normalized to creatinine, was determined in 44 samples from 24 patients with underlying hematologic malignancies and probable, possible or no IA according to current EORTC/MSG criteria and compared to other established biomarkers measured in urine and same-day blood samples. Results TAFC/creatinine sensitivity, specificity, positive and negative likelihood ratio for probable versus no IA (cut-off ≥ 3) were 0.86, 0.88, 6.86, 0.16 per patient. Conclusion For the first time, we provide proof for the occurrence of TAFC in human urine. TAFC/creatinine index determination in urine showed promising results for diagnosis of IA offering the advantages of non-invasive sampling. Sensitivity and specificity were similar as reported for GM determination in serum and bronchoalveolar lavage, the gold standard mycological criterion for IA diagnosis. KW - aspergillosis KW - biomarker KW - diagnosis KW - siderophore KW - urine Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-320939 VL - 78 ER - TY - JOUR A1 - Heimann, Sebastian M. A1 - Penack, Olaf A1 - Heinz, Werner J. A1 - Rachow, Tobias A1 - Egerer, Gerlinde A1 - Kessel, Johanna A1 - Claßen, Annika Y. A1 - Vehreschild, Jörg Janne T1 - Intravenous and tablet formulation of posaconazole in antifungal therapy and prophylaxis: A retrospective, non-interventional, multicenter analysis of hematological patients treated in tertiary-care hospitals JF - International Journal of Infectious Diseases N2 - Objectives Novel formulations (gastro-resistant tablet and intravenous solution) of posaconazole (POS) have been approved in prophylaxis and therapy of invasive fungal diseases (IFDs). Study aim was to analyze treatment strategies and clinical effectiveness. Methods We set up a web-based registry on www.ClinicalSurveys.net for documentation of comprehensive data of patients who received novel POS formulations. Data analysis was split into two groups of patients who received novel POS formulations for antifungal prophylaxis (posaconazole prophylaxis group) and antifungal therapy (posaconazole therapy group), respectively. Results Overall, 180 patients (151 in the posaconazole prophylaxis group and 29 in the posaconazole therapy group) from six German tertiary care centers and hospitalized between 05/2014 – 03/2016 were observed. Median age was 58 years (range: 19 – 77 years) and the most common risk factor for IFD was chemotherapy (n = 136; 76%). In the posaconazole prophylaxis group and posaconazole therapy group, median POS serum levels at steady-state were 1,068 μg/L (IQR 573–1,498 μg/L) and 904 μg/L (IQR 728–1,550 μg/L), respectively (P = 0.776). During antifungal prophylaxis with POS, nine (6%) probable/proven fungal breakthroughs were reported and overall survival rate of hospitalization was 86%. The median overall duration of POS therapy was 18 days (IQR: 7 – 23 days). Fourteen patients (48%) had progressive IFD under POS therapy, of these five patients (36%) died related to or likely related to IFD. Conclusions Our study demonstrates clinical effectiveness of antifungal prophylaxis with novel POS formulations. In patients treated for possible/probable/proven IFD, we observed considerable mortality in patients receiving salvage treatment and with infections due to rare fungal species. KW - invasive fungal infection KW - neutropenia KW - posaconazole serum level KW - clinical effectiveness KW - high-risk patient Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319567 VL - 83 ER - TY - JOUR A1 - Storey, Benjamin C. A1 - Staplin, Natalie A1 - Haynes, Richard A1 - Reith, Christina A1 - Emberson, Jonathan A1 - Herrington, William G. A1 - Wheeler, David C. A1 - Walker, Robert A1 - Fellström, Bengt A1 - Wanner, Christoph A1 - Landray, Martin J. A1 - Baigent, Colin T1 - Lowering LDL cholesterol reduces cardiovascular risk independently of presence of inflammation JF - Kidney International N2 - Markers of inflammation, including plasma C-reactive protein (CRP), are associated with an increased risk of cardiovascular disease, and it has been suggested that this association is causal. However, the relationship between inflammation and cardiovascular disease has not been extensively studied in patients with chronic kidney disease. To evaluate this, we used data from the Study of Heart and Renal Protection (SHARP) to assess associations between circulating CRP and LDL cholesterol levels and the risk of vascular and non-vascular outcomes. Major vascular events were defined as nonfatal myocardial infarction, cardiac death, stroke or arterial revascularization, with an expanded outcome of vascular events of any type. Higher baseline CRP was associated with an increased risk of major vascular events (hazard ratio per 3x increase 1.28; 95% confidence interval 1.19-1.38). Higher baseline LDL cholesterol was also associated with an increased risk of major vascular events (hazard ratio per 0.6 mmol/L higher LDL cholesterol; 1.14, 1.06-1.22). Higher baseline CRP was associated with an increased risk of a range of non-vascular events (1.16, 1.12-1.21), but there was a weak inverse association between baseline LDL cholesterol and non-vascular events (0.96, 0.92-0.99). The efficacy of lowering LDL cholesterol with simvastatin/ezetimibe on major vascular events, in the randomized comparison, was similar irrespective of CRP concentration at baseline. Thus, decisions to offer statin-based therapy to patients with chronic kidney disease should continue to be guided by their absolute risk of atherosclerotic events. Estimation of such risk may include plasma biomarkers of inflammation, but there is no evidence that the relative beneficial effects of reducing LDL cholesterol depends on plasma CRP concentration. KW - C-reactive protein KW - inflammation KW - LDL cholesterol KW - randomized trials KW - vascular disease Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-240067 VL - 93 ER - TY - JOUR A1 - Zhang, Zishuai A1 - Ye, Siyu A1 - Gbureck, Uwe A1 - Barralet, Jake E. A1 - Merle, Géraldine T1 - Cavitation Mediated 3D Microstructured Architectures from Nanocarbon JF - Advanced Functional Materials N2 - Here, the formation of high surface area microscale assemblies of nanocarbon through phosphate and ultrasound cavitation treatment is reported. Despite high conductivity and large surface area, potential health and safety concerns limit the use of nanocarbon and add challenges to handling. Previously, it is shown that phosphate ultrasonic bonding is ineffective for organic materials but in this study, it is found that by a preliminary oxidizing treatment, several carbons can be readily assembled from xerogels. Assembling nanocarbon into microparticles can usually require a binder or surfactants, which can reduce surface area or conductivity and generate a low microsphere yield. Carbon nanotube microspheres are nitrogen-doped and flower-like nanostructured Pt deposited on their surface, and finally showcased as efficient cathode electrocatalysts for the oxygen reduction reaction (half-wave potential 0.78 V vs reversible hydrogen electrode) and methanol oxidation (417 mA mg−1). In particular, no significant degradation of the catalysts is detected after 12 000 cycles (26.6 h). These results indicate the potential of this multimaterial assembly method and open a new way to improve handling of nanoscale materials. KW - carbon nanotube microspheres KW - cavitation KW - oxygen reduction reaction KW - platinum nanostructures Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233926 VL - 28 ER - TY - JOUR A1 - Mueller, Dolores A1 - Jung, Kathrin A1 - Winter, Manuel A1 - Rogoll, Dorothee A1 - Melcher, Ralph A1 - Kulozik, Ulrich A1 - Schwarz, Karin A1 - Richling, Elke T1 - Encapsulation of anthocyanins from bilberries – Effects on bioavailability and intestinal accessibility in humans JF - Food Chemistry N2 - Anthocyanins are flavonoids that have been suggested to provide beneficial health effects. The biological activity of anthocyanins is influenced by their pharmacokinetic properties, but anthocyanins are associated with limited bioavailability in humans. In the presented study, we investigated how the encapsulation of bilberry extract (BE), a source of anthocyanins, with either whey protein or citrus pectin influences the bioavailability and intestinal accessibility of anthocyanins in humans. We performed an intervention study that analyzed anthocyanins and their degradation products in the urine, plasma, and ileal effluent of healthy volunteers and ileostomists (subjects without an intact colon). We were able to show, that whey protein encapsulation modulated short-term bioavailability and that citrus pectin encapsulation increased intestinal accessibility during passage through the small intestine and modulated the formation of the degradation product phloroglucinol aldehyde (PGAL) in human plasma. KW - anthocyanins KW - encapsulation KW - human intervention KW - bioavailability KW - phloroglucinol aldehyde Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224247 VL - 248 ER - TY - JOUR A1 - McMaster, Rebecca A1 - Hoefner, Christiane A1 - Hrynevich, Andrei A1 - Blum, Carina A1 - Wiesner, Miriam A1 - Wittmann, Katharina A1 - Dargaville, Tim R. A1 - Bauer-Kreisel, Petra A1 - Groll, Jürgen A1 - Dalton, Paul D. A1 - Blunk, Torsten T1 - Tailored Melt Electrowritten Scaffolds for the Generation of Sheet-Like Tissue Constructs from Multicellular Spheroids JF - Advanced Healthcare Materials N2 - Melt electrowriting (MEW) is an additive manufacturing technology that is recently used to fabricate voluminous scaffolds for biomedical applications. In this study, MEW is adapted for the seeding of multicellular spheroids, which permits the easy handling as a single sheet-like tissue-scaffold construct. Spheroids are made from adipose-derived stromal cells (ASCs). Poly(ε-caprolactone) is processed via MEW into scaffolds with box-structured pores, readily tailorable to spheroid size, using 13–15 µm diameter fibers. Two 7–8 µm diameter “catching fibers” near the bottom of the scaffold are threaded through each pore (360 and 380 µm) to prevent loss of spheroids during seeding. Cell viability remains high during the two week culture period, while the differentiation of ASCs into the adipogenic lineage is induced. Subsequent sectioning and staining of the spheroid-scaffold construct can be readily performed and accumulated lipid droplets are observed, while upregulation of molecular markers associated with successful differentiation is demonstrated. Tailoring MEW scaffolds with pores allows the simultaneous seeding of high numbers of spheroids at a time into a construct that can be handled in culture and may be readily transferred to other sites for use as implants or tissue models. KW - 3D printing KW - additive manufacturing KW - adipose tissue engineering Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223921 VL - 8 ER - TY - JOUR A1 - Dietz, Maximilian A1 - Johnson, Alice A1 - Martínez-Martínez, Antonio A1 - Weller, Andrew S. T1 - The [Rh(Xantphos)]+ catalyzed hydroboration of diphenylacetylene using trimethylamine-borane JF - Inorganica Chimica Acta N2 - The rhodium(I) complex [Rh(κ3-P,O,P-Xantphos)(η2-PhC≡CPh)][BArF4] (ArF = 3,5-(CF3)2C6H4) is an effective catalyst for the cis-selective hydroboration of the alkyne diphenylacetylene using the amine-borane H3B·NMe3. Detailed mechanistic studies, that include initial rate measurements, full simulation of temporal profiles for a variety of catalyst and substrate concentrations, and speciation experiments, suggest a mechanism that involves initial coordination of alkyne and a saturation kinetics regime for amine-borane binding. The solid-state molecular structure of a model complex that probes the proposed resting state is also reported, [Rh(κ3-P,O,P-Xantphos)(NCMe)(η2-PhC≡CPh)][BArF4]. KW - rhodium KW - hydroboration KW - amine borane KW - mechanism Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-225352 VL - 491 ER - TY - JOUR A1 - Selcuk, Nalan Alan A1 - Toklu, Turkay A1 - Beykan, Seval A1 - Karaaslan, Serife Ipek T1 - Evaluation of the dosimetry approaches in ablation treatment of thyroid cancer JF - Journal of Applied Clinical Medical Physics N2 - In this study, we aimed to evaluate dosimetric approaches in ablation treatment of Differentiated Thyroid Carcinoma (DTC) without interrupting the clinical routine. Prior to therapy, 10.7 MBq 131I in average was orally given to 24 patients suffering from DTC. MIRD formalism was used for dosimetric calculations. For blood and bone marrow dosimetry, blood samples and whole-body counts were collected at 2, 24, 72, and 120 h after I-131 administration. For remnant tissue dosimetry, uptake measurements were performed at the same time intervals. To estimate the remnant volume, anterior and lateral planar gamma camera images were acquired with a reference source within the field of view at 24 h after I-131 administration. Ultrasound imaging was also performed. Treatment activities determined with the fixed activity method were administered to the patients. Secondary cancer risk relative to applied therapy was evaluated for dosimetric approaches. The average dose to blood and bone marrow were determined as 0.15 ± 0.04 and 0.11 ± 0.04 Gy/GBq, respectively. The average remnant tissue dose was 0.58 ± 0.52 Gy/MBq and the corresponding required activity to ablate the remnant was approximately 1.3 GBq of 131I. A strong correlation between 24th-hour uptake and time-integrated activity coefficient values was obtained. Compared to fixed activity method, approximately five times higher secondary cancer risk was determined in bone marrow dosimetry, while the risk was about three times lower in lesion-based dosimetry. KW - bone marrow dosimetry KW - remnant tissue dosimetry KW - thyroid ablation treatment Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-235882 VL - 19 ER - TY - JOUR A1 - Grebinyk, Anna A1 - Grebinyk, Sergii A1 - Prylutska, Svitlana A1 - Ritter, Uwe A1 - Matyshevska, Olga A1 - Dandekar, Thomas A1 - Frohme, Marcus T1 - C60 fullerene accumulation in human leukemic cells and perspectives of LED-mediated photodynamic therapy JF - Free Radical Biology and Medicine N2 - Recent progress in nanobiotechnology has attracted interest to a biomedical application of the carbon nanostructure C60 fullerene since it possesses a unique structure and versatile biological activity. C60 fullerene potential application in the frame of cancer photodynamic therapy (PDT) relies on rapid development of new light sources as well as on better understanding of the fullerene interaction with cells. The aim of this study was to analyze C60 fullerene effects on human leukemic cells (CCRF-CEM) in combination with high power single chip light-emitting diodes (LEDs) light irradiation of different wavelengths: ultraviolet (UV, 365 nm), violet (405 nm), green (515 nm) and red (632 nm). The time-dependent accumulation of fullerene C60 in CCRF-CEM cells up to 250 ng/106 cells at 24 h with predominant localization within mitochondria was demonstrated with immunocytochemical staining and liquid chromatography mass spectrometry. In a cell viability assay we studied photoexcitation of the accumulated C60 nanostructures with ultraviolet or violet LEDs and could prove that significant phototoxic effects did arise. A less pronounced C60 fullerene phototoxic effect was observed after irradiation with green, and no effect was detected with red light. A C60 fullerene photoactivation with violet light induced substantial ROS generation and apoptotic cell death, confirmed by caspase3/7 activation and plasma membrane phosphatidylserine externalization. Our work proved C60 fullerene ability to induce apoptosis of leukemic cells after photoexcitation with high power single chip 405 nm LED as a light source. This underlined the potential for application of C60 nanostructure as a photosensitizer for anticancer therapy. KW - C-60 fullerene KW - photodanamic therapy KW - LEDs KW - leukemic cells KW - immunocytochemistry KW - HPLC-ESI-MS KW - apoptosis Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228245 VL - 124 ER - TY - THES A1 - Aljasem, Anwar T1 - Der Einfluss des Hepatocyte growth factors auf die PD-L1-Expression in Kopf-Hals-Karzinomen: Die Bedeutung des MAPK-, AKT- und STAT3-Signalwegs T1 - The impact of Hepatocyte growth factor (HGF) on PD-L1 expression in HNSCC: The meaning of MAPK, AKT, and STAT3 signaling pathways N2 - Die zielgerichtete Therapie und die Immuncheckpoint-Inhibitoren haben die Tumortherapie revolutioniert. Während erstere die Tumorzellen gezielt angreift, verhindern letztere die Hemmung des Immunsystems durch Immuncheckpoints, um eine robuste Immunantwort zu erreichen. Zusätzlich ist das Nebenwirkungsprofil bei direktem Vergleich mit der konventionellen Chemotherapie günstiger. Beim HNSCC werden beide Ansätze angewendet. Cetuximab ist ein monoklonaler Antikörper, der sich gegen EGFR, welcher bei HNSCC überexprimiert ist, richtet. Nivolumab und Pembrolizumab richten sich gegen das Immuncheckpoint-Protein PD-1. Nach wie vor sind die Resistenzen, sowohl die initialen als auch die erworbenen, die größte zu überwindende Herausforderung. Aufbauend auf dem Ergebnis vorangegangener Arbeiten, die zeigen konnten, dass HGF über c-MET die Expression des Immuncheckpointliganden PD-L1 steigert, setzt sich diese Arbeit weiter mit den intrazellulären nachgeschalteten Signalwegen nach c-MET Aktivierung auseinander. Dies ist von besonderem Interesse, weil diese Signalwege ebenfalls für die Resistenzentwicklung verantwortlich sein können, zeitgleich können diese im Rahmen der zielgerichteten Therapie gezielt inhibiert werden. Um den HGF-Einfluss auf die intrazellulären Signalwege zu prüfen, wurden vier etablierte HNSCC-Zelllinien herangezogen. Im ersten Teil der Arbeit wurden die 4 HNSCC-abgeleitete Zelllinien mit HGF stimuliert und mittels Western Blot der PD-L1-Anstieg und die Phosphorylierungsänderung der Schlüsselproteine der einzelnen Signalwege nachgewiesen. Daraus ergab sich, dass HGF die MAPK- und PIK3/AKT-Signalwege aktiviert. Während eine kombinierte Blockade des MAPK-Signalwegs den PD-L1-Anstieg vollständig verhindern konnte, hemmte die PIK3/AKT-Blockade den PD-L1-Anstieg nur partiell. Im zweiten Teil wurde mit siRNA der hauptsächlich für den PD-L1-Anstieg zuständige MAPK-Signalweg unterbunden, was mittels quantitativer PCR auf der mRNA-Ebene nachgewiesen werden konnte. Mittels Western Blot konnte entsprechend gezeigt werden, dass der PD-L1-Anstieg trotz HGF-Stimulation bei nicht funktionsfähigem MAPK-Signalweg eingeschränkt war. Weiter wurde der Effekt mit dem Medikament Trametinib, das im Rahmen der zielgerichteten Therapie bei malignem Melanom und NSCLC für die MAPK-Signalweg-Hemmung zugelassen ist, evaluiert. Sowohl im Western Blot als auch in der Durchflusszytometrie konnte bestätigt werden, dass Trametinib den HGF-induzierten Anstieg von PD-L1 signifikant blockiert. Darüber hinaus konnte im Rahmen der Western Blot-Versuche gezeigt werden, dass die Signalwege und die PD-L1-Expression in den Zelllinien unterschiedlich aktiv bzw. hoch waren. Unter den vier Zelllinien zeigte die FaDu-Zelllinie eine erhöhte PI3K/AKT-Aktivität, Detroit562 und SCC9 eine erhöhte MAPK-Aktivität. Die PD-L1- Expression war in der SCC9-Zelllinie am höchsten. Die Arbeit zeigt eine einheitliche Reaktion der HNSCC-Zelllinien auf den Wachstumsfaktor HGF, welcher im Tumormilieu von HNSCC oft in hoher Konzentration vorhanden ist. Neben dem EGFR-Antikörper (Cetuximab) kann eine kombinierte Hemmung entweder von c-MET oder von den nachgeschalteten Signalwegen MAPK und PI3K/AKT bei Resistenzen, Progression oder Unverträglichkeiten eine Möglichkeit für eine wirksamere Therapie von HNSCC darstellen. Ein Screening der Signalwege und deren Aktivierungsmechanismen könnte bei Resistenzen oder bei einem Rezidiv/Progress dazu beitragen, gezielt die alternative Aktivierung zu hemmen und möglicherweise die Wirksamkeit einer Immuncheckpointblockade zu verbessern. N2 - Targeted therapy and immune checkpoint inhibitors have revolutionized tumor therapy. While the former specifically targets tumor cells, the latter prevents inhibitory immune responses via immune checkpoints to achieve a robust immune response. Additionally, the side effect profile is more favorable when directly compared to conventional chemotherapy. Both approaches are approved for the treatment of Head and Neck Squamous Cell Carcinoma (HNSCC). Cetuximab is a monoclonal antibody that targets EGFR, which is overexpressed in HNSCC. Nivolumab and Pembrolizumab target the immune checkpoint protein PD-1. Resistances, both initial and acquired, however, remain significant challenges to overcome. Building on a previous report that HGF upregulates the immune checkpoint ligand PD-L1 expression via the c-MET pathway, this study investigates the intracellular downstream signaling pathways activated by HGF. This is of particular interest because these signaling pathways contribute to resistance development, while at the same time, they can be specifically inhibited in the context of targeted therapy. To examine the influence of HGF on intracellular signaling pathways, four established HNSCC cell lines were utilized. In the first part of the study, the four HNSCC-derived cell lines were stimulated with HGF, and the increase in PD-L1 expression and changes in phosphorylation levels of key proteins in the individual signaling pathways were analyzed using Western Blots. It was found that HGF activates the MAPK and PI3K/AKT pathways. While MAPK inhibition completely blocked the PD-L1 increase, PI3K/AKT inhibition only partially did so after HGF stimulation. In the second part, the MAPK pathway, mainly responsible for the increase in PD-L1, was inhibited using siRNA. Quantitative PCR validated corresponding mRNA levels. Western Blots further showed that the increase in PD-L1 was reduced despite HGF stimulation when the MAPK pathway was non-functional. In line with previous results, inhibiting MAPKs with the drug Trametinib, which is approved as a targeted therapy for malignant melanoma and NSCLC, significantly blocks HGF-induced PD-L1 expression based on Western Blot and flow cytometry analysis. Notably, the activity of the different signaling pathways and the expression levels of PD-L1 vary among the different cell lines, as Western Blot analyses reveal. For instance, while the FaDu cell line manifested high activity in the PI3K/AKT pathway, Detroit562 and SCC9 showed increased MAPK activity. Nonetheless, the SCC9 cell line showed the highest PD-L1 expression level. This study demonstrated that HNSCC cell lines similarly respond to the growth factor HGF, which is frequently present in high concentrations in the tumor microenvironment of HNSCC. Additionally, the inhibition of c-MET and its downstream MAPK or PI3K/AKT pathways in combination with EGFR blockade by Cetuximab may offer a more effective treatment strategy for HNSCC patients in cases of therapeutic resistance, disease progression, or drug intolerance. Screening of the signaling pathways and their activation mechanisms could, in cases of resistance, recurrence, or progression, contribute to the specific inhibition of alternative activation and potentially improve the efficacy of immune checkpoint blockade. KW - Hepatozyten-Wachstumsfaktor KW - MAP-Kinase KW - Plattenepithelcarcinom KW - PD-L1 Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370358 ER - TY - THES A1 - Müller, Nicole T1 - Modellierung klonaler Evolution beim Multiplen Myelom T1 - Modeling Clonal Evolution in Multiple Myeloma N2 - In dieser Arbeit wurde ein modulares Zelllinienmodell zur Visualisierung klonaler Evolutionsmechanismen etabliert. Hierfür wurden unterschiedlich fluoreszierende Proteine (LSSmKate2, EGFP, mTagBFP2) durch Anwendung eines Sleeping Beauty basierten Vektorsystems in unterschiedliche Sublinien der Myelom Zelllinie L363 eingebracht. Diese vier Sublinien beinhalten jeweils eine von drei aus primären Patientenproben gewonnenen Mutationen in IKZF1 (A152T, E170D, R439H) oder den IKZF1 WT. Die Anwendung von immunmodulatorischen Medikamenten (IMiDs) führt zu einer Ubiquitinierung des Transkriptionsfaktors IKZF1 durch die E3-Ubiquitin-Protein-Ligase (CRBN-CUL4). Durch Mutationen in IKZF1 kommt es zu Störungen in diesem Prozess und damit zu einer Überexpression von IKZF1. Dies wirkt sich wachstumsfördert auf die Myelomzellen aus. Die Auswirkungen der einzelnen Mutationen in IKZF1 ist aufgrund dessen ein klinisch relevantes Forschungsthema. In dieser Arbeit wurden jeweils zwei Sublinien mit Zellen des IKZF1 WT und Zellen mit einer IKZF1 Mutation mit jeweils unterschiedlich fluoreszierenden Proteinen markiert. Diese wurden gemeinsam unter Behandlung mit verschiedenen Konzentrationen von Lenalidomid inkubiert. Somit konnte das Selektionsverhalten mittels Durchflusszytometrie-Auswertungen visualisiert werden. Es konnte gezeigt werden, dass die IKZF1 Mutation A152T einen deutlichen Selektionsvorteil für die Myelomzellen darstellt. Bei den IKZF1 Mutationen E170D und R439H konnte kein Selektionsvorteil gegenüber dem IKZF1 WT beobachtet werden. N2 - In this work, a modular cell line model was established to visualize clonal evolutionary mechanisms. Different fluorescent proteins (LSSmKate2, EGFP, mTagBFP2) were introduced into various sublines of the myeloma cell line L363 using a Sleeping Beauty-based vector system. These four sublines each contain one of three mutations in IKZF1 (A152T, E170D, R439H) derived from primary patient samples or the IKZF1 wild type (WT). The application of immunomodulatory drugs (IMiDs) leads to the ubiquitination of the transcription factor IKZF1 by the E3 ubiquitin-protein ligase (CRBN-CUL4). Mutations in IKZF1 disrupt this process, resulting in the overexpression of IKZF1, which promotes the growth of myeloma cells. The effects of individual mutations in IKZF1 are therefore a clinically relevant research topic. In this study, two sublines each with IKZF1 WT cells and cells with an IKZF1 mutation were labeled with different fluorescent proteins. These were incubated together under treatment with various concentrations of lenalidomide. Thus, the selection behavior could be visualized using flow cytometry analyses. It was shown that the IKZF1 mutation A152T provides a clear selective advantage for the myeloma cells. No selective advantage was observed for the IKZF1 E170D and R439H mutations compared to IKZF1 WT. KW - Lenalidomid KW - Plasmozytom KW - IKZF1 KW - klonale Evolution KW - Vektormodell KW - Multiples Myelom Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370818 ER - TY - THES A1 - Lechermeier, Carina T1 - Neuroanatomical and functional evaluation of ADHD candidate genes in the model organism zebrafish (\(Danio\) \(rerio\)) T1 - Neuroanatomische und funktionelle Auswertung von ADHS Kandidatengenen im Modellorganismus Zebrafisch (\(Danio\) \(rerio\)) N2 - Attention deficit hyperactivity disorder (ADHD) is one of the most prevalent developmental disorders, affecting 5.9% children and adolescents and 2.5% adults worldwide. The core characteristics are age-inappropriate levels of hyperactivity, impulsivity and inattention, often accompanied by co-morbidities such as mood and conduct disorders as wells as learning deficits. In the majority of cases, ADHD is caused by an interplay of accumulated genetic and environmental risk factors. Twin studies report a very high heritability of 70–80%, however, common genetic variants in the population only explain a third of the heritability. The rest of the genetic predisposition is composed of rare copy number variations (CNVs) and gene x environment interactions including epigenetic alterations. Through genome wide association (GWAS) and linkage studies a number of likely candidate genes were identified. A handful of them play a role in dopamine or noradrenaline neurotransmitter systems, simultaneously those systems are the main targets of common drug treatment approaches. However, for the majority of candidates the biological function in relation to ADHD is unknown. It is crucial to identify those functions in order to gain a deeper understanding of the pathomechanism and genetic networks potentially responsible for the disorder. This work focuses on the three candidate genes GFOD1, SLC2A3 and LBX1 and their role in the healthy organism as well as in case of ADHD. The neuroanatomy was regarded through expression analysis and various behavioural assays of activity were performed to link alterations on the transcript level to phenotypes associated with the neurodevelopmental disorder. Zebrafish orthologues of the human risk genes were identified and extensive temporal and spacial expression characterisation performed via RNA in situ hybridisation. Through morpholino derived knock-down and mRNA overexpression zebrafish models with subsequent behavioural analysis, both hyper- and hypoactive phenotypes were discovered. Additional expression analysis through double in situ hybridisation revealed a co-localisation during zebrafish neurodevelopment of each gfod1 and slc2a3a together with gad1b, a marker for GABAergic neurons. Interestingly, both risk genes have previously been associated with glucose homeostasis and energy metabolism, which when disrupted could lead to alterations in signal transduction and neuron survival. Likewise, Lbx1 plays a pivotal role in GABAergic versus glutamatergic neuron specification during spinal cord and hindbrain development in mice and chicken. Preliminary results of this work suggest a similar role in zebrafish. Taken together, those findings on the one hand represent a sturdy basis to con- tinue studies of the function of the genes and on the other hand open up the opportunity to investigate novel aspects of ADHD research by exploring the role of the GABAergic neurotransmitter system or the connection between energy metabolism and psychiatric disorders. N2 - Die Aufmerksamkeitsdefizit-/Hyperaktivitätsstörung (ADHS) ist eine der am weitesten verbreiteten Entwicklungsstörungen, davon sind 5,9% Kinder und Jugendliche und 2,5% Erwachsene weltweit betroffen. Die Kernsymptome sind altersunangemessene Hyperaktivität, Impulsivität und Unaufmerksamkeit, oft begleitet von Begleiterkrankungen wie emotionale Dysregulation oder Verhal- tensauffälligkeiten sowie Lerndefiziten. In den meisten Fällen wird ADHS durch ein Zusammenspiel von angehäuften genetischen und umweltbedingten Risikofaktoren verursacht. Durch Zwillingsstudien gelang man zu einer errechneten Erblichkeit von 70–80%, jedoch erklären häufig auftretende genetische Varianten in der Bevölkerung nur ein Drittel der Erblichkeit. Der Rest der genetischen Veranlagung setzt sich aus seltenen Kopienzahlvariationen (CNV) und Interaktionen von Gen x Umwelt, einschließlich epigenetischer Veränderungen, zusammen. Durch genomweite Assoziationsstudien (GWAS) und Kopplungsanalysen wurden eine Reihe von wahrscheinlichen Kandidatengenen identifiziert. Eine Handvoll von ihnen spielen eine Rolle in den Dopamin oder Noradrenalin Neurotransmittersystemen. Diese Systeme sind gleichzeitig die Hauptangriffspunkte der gängigsten Medikamente, die zur Behandlung von ADHS eingesetzt werden. Allerdings ist für die Mehrheit der Kandidatengene die biologische Funktion in Bezug auf ADHS unbekannt. Es ist essentiell diese Funktionen zu identifizieren um ein tieferes Verständnis der Ätiopathogenese und der genetische Netzwerke, die möglicherweise für die Störung verantwortlich sind, zu erlangen. Diese Arbeit konzentriert sich auf die drei Kandidatengene GFOD1, SLC2A3 und LBX1 und ihre Rolle im gesunden Organismus sowie während ADHS. Die Neuroanatomie wurde durch Expressionsanalyse betrachtet und verschiedene aktivitätsbasierte Verhaltensessays wurden durchgeführt, um Veränderungen auf Transkriptebene mit den zugehörigen Phänotypen der neurologischen Entwick- lungsstörung in Verbindung zu bringen. Zebrafischorthologe der menschlichen Kandidatengene wurden identifiziert und umfangreiche zeitliche und räumli- che Expressionsanalysen via RNA in situ Hybridisierung durchgeführt. Durch Morpholino-Knockdown und mRNA-Überexpressions Zebrafischmodelle mit anschließender Verhaltensanalyse wurden sowohl hyper- als auch hypoaktive Phänotypen entdeckt. Eine zusätzliche Expressionsanalyse durch doppelte in situ Hybridisierung ergab eine Kolokalisierung während der Zebrafischneuroentwicklung von jeweils gfod1 und slc2a3a zusammen mit gad1b, einem Marker für GABAerge Neuronen. Interessanterweise wurden beide Risikogene zuvor mit der Glukosehomöostase und dem Energiestoffwechsel in Verbindung gebracht, die, wenn sie gestört werden, zu Veränderungen der Signalübertragung und der Lebensdauer von Neuronen führen können. Desgleichen spielt Lbx1 eine entscheidende Rolle bei der Spezifikation von GABAergen versus glutamatergenen Neuronen während der Entwicklung des Rückenmarks in der Wirbelsäule und im Hinterhirn von Mäusen und Hühnern. Vorläufige Ergebnisse dieser Arbeit deuten auf eine ähnliche Rolle beim Zebrafisch hin. Zusammengenommen stellen diese Erkenntnisse einerseits eine solide Grundlage für weitere Untersuchungen zur Funktion der Gene dar, andererseits eröffnet sich daraus die Möglichkeit neue Aspekte der ADHS-Forschung zu untersuchen, bei denen der Fokus auf der Rolle des GABAergen Neurotransmittersystems oder der Beziehung zwischen Energiestoffwechsel und psychiatrischen Erkrankungen liegt. KW - Aufmerksamkeitsdefizit-Syndrom KW - Zebrabärbling KW - ADHD KW - zebrafish KW - genes KW - behaviour KW - ADHS Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-371084 ER - TY - THES A1 - Rode, Stefan T1 - Automated resummation of electroweak Sudakov logarithms in diboson production T1 - Automatisierte Resummierung elektroschwacher Sudakov-Logarithmen in Vektorboson-Paarproduktion N2 - The present thesis is concerned with the automated computation of integrated and differential cross sections of diboson production in proton–proton and electron–positron collisions at very high energies, including a resummation of electroweak Sudakov logarithms to all orders in the fine-structure constant using soft–collinear effective theory. The search for new physics at future colliders such as the FCC–hh or the CLIC requires precise predictions for scattering cross sections from the theoretical high-energy physics com- munity. Electroweak Sudakov logarithms, which currently limit the accuracy of predictions in the high-energy tails of differential distributions for LHC-like energies, are known to destroy the convergence behaviour of the fixed-order perturbative series, once sufficiently high energies are considered. To resum these large corrections, soft–collinear effective theory has been applied to simple processes, which permits analytic calculations. Within this work, we present an automated computation within a Monte Carlo integration framework, thus facilitating the computation of fully differential cross section to complicated processes. This requires the use of the Catani– Seymour subtraction algorithm to treat the occurring infrared divergences. The machinery is applied to all diboson processes with intermediate weak gauge bosons, including the photon- induced W+ W− -production channel. To this end we carefully study the validity of the necessary assumptions such as the double- pole approximation and estimate the order of magnitude of neglected effects. Especially the non-doubly-resonant contributions turn out to be sizeable in several interesting phase-space regions. For lepton collisions at 3 TeV we obtain the integrated cross sections of W-pair and Z-pair production to be shifted by more than 20% with respect to the Born value, owing to the resum- mation of the leading-logarithmic corrections These effects are partly cancelled by subleading effects. For proton–proton collisions at √ s = 100 TeV we observe sizeable resummation effects in the high-energy tails, while the integrated cross sections are dominated by interactions, for which soft–collinear effective theory is not applicable. N2 - Das Thema ist der vorliegenen Arbeit ist die automatisierte Berechnung differenzieller und integrierter Wirkungsquerschnitte der Paarerzeugung schwerer Eichbosonen bei sehr hohen Streuenergien mit Resummierung der auftretenden elektroschwachen Sudakov-Logarithmen zu allen Ordnungen in der Feinstrukturkonstanten mittels Soft-Collinear Effective Theory. Die Suche nach Physik jenseits des Standardmodells an zukunftigen Teilchenbeschleunigern wie dem FCC oder dem CLIC erfordert hochpräzise Voraussagen fur Streuquerschnitte seitens der theoretischen Physik. Es ist seit langem bekannt, dass elektroschwache Sudakov-Logarithmen, die bereits gegenwärtig die Genauigkeit der Voraussagen in den Hochenergieschwänzen von Verteilungen limitieren, die Konvergenz der konventionellen Störungsreihen vollkommen zunichte machen, wenn hinreichend hohe Energien erreicht werden. Mittels Soft-Collinear Effective Theory wurden diese Logarithmen bereits in der Vergangenheit in einfachen Prozessen, die eine analytische Behandlung erlauben, resummiert. Im Rahmen dieser Arbeit wurden diese Methoden in ein Monte-Carlo-Integrationsprogramm implementiert, um somit vollständig differenzielle Vorhersagen präsentieren zu können. Dies erfordert die Behandlung von Infrarotdivergenzen mit Hilfe des Catani-Seymour-Algorithmus. Mit diesen Werkzeugen wurden resummierte Streuquerschnitte fur verschiedene Vektorboson-Paarproduktionsprozesse berechnet, u.a. fur den Photon-Photon-induzierten Produktionskanal zur W-Boson-Paarproduktion. Auf dem Weg dorthin sind verschiedene vereinfachende Annahmen notwendig, deren Gultigkeit im Rahmen dieser Arbeit ebenfalls getestet wurde, so z.B. die Qualität der Doppelpolnäherung. Des weiteren wurden Größenordnungen vernachlässigter Effekte abgeschätzt. Dabei haben sich vor allem nicht doppelt resonante Beiträge in bestimmten Phasenraumregionen als beträchtlich herausgestellt. Der Resummationseffekt der fuhrend logarithmischen Korrekturen verschiebt die integrierten Paarproduktionsstreuquerschnitte um mehr als 20% bezogen auf den Bornstreuquerschnitt im Falle von Leptonkollisionen bei einer Schwerpunktsenergie von 3 TeV. Diese Effekte werden allerdings teilweise von nicht-führenden Beiträgen kompensiert. Fur Proton-Proton-Kollisionen bei √ s = 100 TeV finden wir deutliche Resummationseffekte in allen Hochenergieschwänzen, während die integrierten Wirkungsquerschnitte von Phasenraumregionen dominiert werden, in denen Soft-Collinear Effective Theory nicht anwendbar ist. KW - High-energy physics Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-371060 ER - TY - JOUR A1 - Gorlova, Anna A1 - Pavlov, Dmitrii A1 - Anthony, Daniel C. A1 - Ponomarev, Eugene D. A1 - Sambon, Margaux A1 - Proshin, Andrey A1 - Shafarevich, Igor A1 - Babaevskaya, Diana A1 - Lesch, Klaus-Peter A1 - Bettendorff, Lucien A1 - Strekalova, Tatyana T1 - Thiamine and benfotiamine counteract ultrasound-induced aggression, normalize AMPA receptor expression and plasticity markers, and reduce oxidative stress in mice JF - Neuropharmacology N2 - The negative societal impacts associated with the increasing prevalence of violence and aggression is increasing, and, with this rise, is the need to understand the molecular and cellular changes that underpin ultrasound-induced aggressive behavior. In mice, stress-induced aggression is known to alter AMPA receptor subunit expression, plasticity markers, and oxidative stress within the brain. Here, we induced aggression in BALB/c mice using chronic ultrasound exposure and examined the impact of the psychoactive anti-oxidant compounds thiamine (vitamin B1), and its derivative benfotiamine, on AMPA receptor subunit expression, established plasticity markers, and oxidative stress. The administration of thiamine or benfotiamine (200 mg/kg/day) in drinking water decreased aggressive behavior following 3-weeks of ultrasound exposure and benfotiamine, reduced floating behavior in the swim test. The vehicle-treated ultrasound-exposed mice exhibited increases in protein carbonyl and total glutathione, altered AMPA receptor subunits expression, and decreased expression of plasticity markers. These ultrasound-induced effects were ameliorated by thiamine and benfotiamine treatment; in particular both antioxidants were able to reverse ultrasound-induced changes in GluA1 and GluA2 subunit expression, and, within the prefrontal cortex, significantly reversed the changes in protein carbonyl and polysialylated form of neural cell adhesion molecule (PSA-NCAM) expression levels. Benfotiamine was usually more efficacious than thiamine. Thus, the thiamine compounds were able to counteract ultrasound-induced aggression, which was accompanied by the normalization of markers that have been showed to be associated with ultrasound-induced aggression. These commonly used, orally-active compounds may have considerable potential for use in the control of aggression within the community. This article is part of the Special Issue entitled ‘Current status of the neurobiology of aggression and impulsivity’. KW - aggression KW - emotional stress KW - brain oxidative stress KW - plasticity KW - thiamine KW - mice Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227439 VL - 156 ER -