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 - TY - JOUR A1 - Hedrich, Rainer A1 - Mueller, Thomas D. A1 - Becker, Dirk A1 - Marten, Irene T1 - Structure and Function of TPC1 Vacuole SV Channel Gains Shape JF - Molecular Plant N2 - Plants and animals in endosomes operate TPC1/SV-type cation channels. All plants harbor at least one TPC1 gene. Although the encoded SV channel was firstly discovered in the plant vacuole membrane two decades ago, its biological function has remained enigmatic. Recently, the structure of a plant TPC1/SV channel protein was determined. Insights into the 3D topology has now guided site-directed mutation approaches, enabling structure–function analyses of TPC1/SV channels to shed new light on earlier findings. Fou2 plants carrying a hyperactive mutant form of TPC1 develop wounding stress phenotypes. Recent studies with fou2 and mutants that lack functional TPC1 have revealed atypical features in local and long-distance stress signaling, providing new access to the previously mysterious biology of this vacuolar cation channel type in planta. KW - Ca2+ sensors KW - TPC1/SV channel KW - vacuole membrane voltage KW - voltage sensor Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228046 VL - 11 ER - TY - JOUR A1 - Verheijen, Bert M. A1 - Stevens, Jo A. A. A1 - Gentier, Romina J. G. A1 - van't Hekke, Christian D. A1 - van den Hove, Daniel L. A. A1 - Hermes, Denise J. H. P. A1 - Steinbusch, Harry W. M. A1 - Ruijter, Jan M. A1 - Grimm, Marcus O. W. A1 - Haupenthal, Viola J. A1 - Annaert, Wim A1 - Hartmann, Tobias A1 - van Leeuwen, Fred W. T1 - Paradoxical effects of mutant ubiquitin on Aβ plaque formation in an Alzheimer mouse model JF - Neurobiology of Aging N2 - Amyloid-β (Aβ) plaques are a prominent pathological hallmark of Alzheimer's disease (AD). They consist of aggregated Aβ peptides, which are generated through sequential proteolytic processing of the transmembrane protein amyloid precursor protein (APP) and several Aβ-associated factors. Efficient clearance of Aβ from the brain is thought to be important to prevent the development and progression of AD. The ubiquitin-proteasome system (UPS) is one of the major pathways for protein breakdown in cells and it has been suggested that impaired UPS-mediated removal of protein aggregates could play an important role in the pathogenesis of AD. To study the effects of an impaired UPS on Aβ pathology in vivo, transgenic APPSwe/PS1ΔE9 mice (APPPS1) were crossed with transgenic mice expressing mutant ubiquitin (UBB+1), a protein-based inhibitor of the UPS. Surprisingly, the APPPS1/UBB+1 crossbreed showed a remarkable decrease in Aβ plaque load during aging. Further analysis showed that UBB+1 expression transiently restored PS1-NTF expression and γ-secretase activity in APPPS1 mice. Concurrently, UBB+1 decreased levels of β-APP-CTF, which is a γ-secretase substrate. Although UBB+1 reduced Aβ pathology in APPPS1 mice, it did not improve the behavioral deficits in these animals. KW - mutant ubiquitin KW - ubiquitin-proteasome system KW - γ-secretase KW - amyloid-β KW - behavior KW - Alzheimer's disease Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233185 VL - 72 ER - TY - THES A1 - Liu, Yang T1 - Predictions for Composite Higgs Models Using Gauge/Gravity Duality T1 - Vorhersagen für zusammengesetzte Higgs-Modelle unter Verwendung der Eich-/Gravitationsdualität N2 - This thesis is dedicated to construct a non-abelian holographic dynamical minimal composite Higgs model. We first build a non-abelian bottom-up AdS/YM model that can explain the QCD meson spectrum well. The model is made non-abelian by considering non-abelian DBI action in the top-down model. We then change the dual theory from the QCD to the minimal composite Higgs model U (4)/Sp(4). By adding a second explicit U (4) → Sp(4) breaking through the NJL interaction at the boundary, we managed to construct a composite Higgs phase and a technicolor phase in this model. The transition between the two phases is also realized, which is controlled by the NJL coupling. This thesis is based on the works [1, 2]. N2 - Diese Arbeit konstruiert ein nicht-abelsches holographisches dynamisches minimales Composite-Higgs-Modell. Wir erstellen zunächst ein nicht-abelsches Bottom-up-AdS/YM-Modell, das das QCD-Mesonenspektrum gut erklären kann. Das Modell ist nicht-abelsch, da die nicht-abelsche DBI-Wirkung im Top-Down-Modell berücksichtigt wird. Anschließend ändern wir die duale Theorie von der QCD auf das minimale Composite-Higgs-Modell U (4)/Sp(4). Durch das Hinzufügen einer zweiten expliziten Brechung U (4) → Sp(4), das die NJL-Wechselwirkung an der Grenze durchbricht, konstruierten wir in diesem Modell eine Composite-Higgs-Phase und eine Technicolor-Phase. Auch der Übergang zwischen den beiden Phasen wird realisiert, welcher durch die NJL-Kopplung gesteuert wird. Diese Arbeit basiert auf den Arbeiten [1, 2]. KW - Composite Higgs KW - Gauge/gravity duality KW - holographic model KW - Higgs-Modell Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370833 ER - TY - THES A1 - Fuhl, Lucas T1 - Photolumineszenzmikroskopie und -spektroskopie endohedraler Farbstoffe in Bornitridnanoröhren T1 - Photoluminescence microscopy and spectroscopy of endohedral dyes in boron nitride nanotubes N2 - Im Rahmen der vorliegenden Dissertation wurde untersucht, wie die Einkapselung organischer Farbstoffmoleküle in Bornitridnanoröhren (BNNTs) die photophysikalischen Eigenschaften der Fluorophore beeinflusst. Als Farbstoffe wurden hierbei alpha-Quaterthiophen (4T), alpha-Sexithiophen (6T), alpha-Octithiophen (8T) sowie Nilrot (NR) ausgewählt. Die eingesetzten BNNTs besitzen einen nominellen Durchmesser von \(5 \pm 2\)nm. Für die Charakterisierung der reinen Farbstoffe und der hybriden Systeme aus Farbstoff und Nanoröhre kam ein Laboraufbau zum Einsatz, der neben Absorptions- und Photolumineszenz (PL)-Spektroskopie auch PL-Mikroskopie ermöglicht. Zusätzlich lässt sich damit auch eine zeitaufgelöste Untersuchung der PL (engl. time correlated single photon counting, TCSPC) im Ensemble und an einzelnen, separierten Nano-Objekten (mit Farbstoff gefüllte BNNTs) umsetzen. In Kapitel 5 wurden zunächst die freien Farbstoffe in Lösung charakterisiert. Es hat sich gezeigt, dass sowohl 4T als auch NR im verwendeten Lösemittel Dimethylformamid (DMF) löslich sind, wohingegen 6T und 8T hier eine geringere Löslichkeit zeigen. Die unterschiedlichen Verläufe der konzentrationsabhängigen PL-Spektren für 4T und 6T in DMF lassen sich vermutlich auf diesen Löslichkeitsunterschied zurückführen. Zudem wurden Extinktionskoeffizienten für 4T und NR mittels konzentrationsabhängiger Absorptionsspektren bestimmt und es zeigte sich eine gute Übereinstimmung mit der Literatur. Für 6T und 8T war eine Bestimmung aufgrund der geringen Löslichkeit nicht möglich, weshalb auf Literaturwerte zurückgegriffen wurde oder diese extrapoliert wurden (8T). In Kapitel 6 erfolgte die detaillierte Charakterisierung der mit Oligothiophenen gefüllten BNNTs. Die Befüllung wurde dabei im Wesentlichen nach einem von C. Allard publizierten Verfahren durchgeführt und auf die zusätzlichen Fluorophore 4T, 8T und NR übertragen. Für Messungen mittels UV-Vis-Spektroskopie in Lösung bzw. Dispersion hat sich beim Farbstoff 6T gezeigt, dass sich das Absorptionsmaximum von 407nm (freies 6T) hin zu 506nm (6T@BNNT) verschiebt. Ursache hierfür ist vermutlich die Bildung von J-Aggregaten im Inneren der Röhren. Die entsprechenden PL-Spektren von freiem 6T und dem Hybridsystem zeigen dabei keine signifikanten Unterschiede. Für konzentrationsabhängige PL-Spektren von 6T@BNNT ergibt sich (anders als bei freiem 6T in DMF) keine Änderung des Verlaufs der Kurven, was als ein Indiz für eine erfolgreiche Einkapselung gedeutet werden kann. Durch Kombination von Rasterkraft- und PL-Mikroskopie konnten die Außendurchmesser von einzelnen 6T@BNNT Objekten ermittelt und in direkten Zusammenhang mit deren photophysikalischen Eigenschaften gebracht werden. Bei einer Analyse der Polarisation des Emissionslichtes von 6T@BNNT in Abhängigkeit des Außendurchmessers ließ sich jedoch keine klare Korrelation zwischen Struktur und Emissionscharakteristiken erkennen. Diese Beobachtung lässt sich vermutlich dadurch erklären, dass mit Hilfe der Rasterkraftmikroskopie lediglich der Außendurchmesser der (teils mehrwandigen) BNNTs bestimmt werden kann. Die entscheidende Größe an dieser Stelle ist allerdings der innere Durchmesser der BNNTs, welcher die Ausrichtung und damit auch die Polarisation der Farbstoffmoleküle beeinflusst. Ein Vergleich des mittleren maximalen Polarisationsgrades der jeweiligen Hybridsysteme hat gezeigt, dass 4T@BNNT den geringsten und 6T@BNNT mit den höchsten Wert aufweist. Dies bestätigt die Annahme, dass mit zunehmender Moleküllänge die Polarisation, aufgrund des höheren Templat-Effektes der Röhre, zunimmt. 8T@BNNT liegt zwischen den beiden anderen Werten, was dieser Annahme widerspricht. Der mittlere Verkippungswinkel der eingekapselten Farbstoffmoleküle gegenüber der Röhrenachse liegt für 4T@BNNT bei etwa 16° und ist damit etwas größer als derjenige von 6T@BNNT. Somit zeigt sich auch hier, dass kürzere Moleküle mehr sterische Freiheitsgerade im Innern der Röhren besitzen. Für 8T@BNNT liegt der Winkel bei ca. 28° und widerspricht abermals der Annahme. TCSPC-Messungen an freien Oligothiophen-Farbstoffen sowie an den hybriden Systemen zeigten, dass die Fluoreszenzlebensdauer \(\tau\) für 4T und 6T (jeweils in DMF) infolge der Einkapselung deutlich zunimmt wenn die Hybridsysteme ebenfalls in DMF dispergiert sind. Die ermittelten Werte für \(\tau\) der separierten Nanoobjekte lagen für 4T@BNNT und 6T@BNNT unterhalb der entsprechenden in DMF. Für 8T bzw. 8T@BNNT ergab sich eine deutlich kürzerer Lebensdauer der separierten Nanoobjekte im Vergleich zum freien Farbstoff in kolloidaler Suspension. Ein erster Ansatz, um den zugrundeliegende Mechanismus aufzuklären, bestand darin, die TCSPC-Spektren (für 6T in DMF und 6T@BNNT in DMF) hinsichtlich der einzelnen Zerfallskanäle zu analysieren. Die erhaltenen Ergebnisse deuteten darauf hin, dass bei freiem 6T in DMF andere Zerfallskanäle dominieren als beim Hybridsystem 6T@BNNT (in DMF). Eine Korrelation der Fluorezenslebensdauer von 6T@BNNT vom äußeren Durchmesser der Nanoröhren zeigte keinen eindeutigen Zusammenhang. Die Charakterisierung von Nilrot bzw. NR@BNNT (analog zu den Oligothiophenen) erfolgte in Kapitel 4. Auch hier zeigte sich eine Verschiebung des PL-Spektrums des Fluorophores durch die Einkapselung in die BNNTs. Allerdings ist das PL-Spektrum des Hybridsystems (NR@BNNT) um etwa 20nm hypsochrom verschoben. Nilrot ist in der Literatur zudem als Nanosonde zur Ermittlung der Permittivität des Lösemittels bzw. der Umgebung bekannt. Dies erlaubte eine Abschätzung der relativen Permittivät im Inneren der BNNTs. Der ermittelte Wert von ca. 4 für ein isoliertes NR@BNNT Objekt deutet auf eine relativ unpolare Umgebung im Röhreninneren hin. Zum Vergleich dazu, liegt der Wert von freiem NR in DMF bei 47, was die relativ hohe Polarität von DMF bestätigt. Der ermittelte Wert für die mittlere maximale Polarisation lag leicht über dem der hybriden Systeme aus Oligothiophenen und Nanoröhren. Für die Auslenkung der NR-Moleküle gegenüber der Röhrenachse ergab sich ein Winkel von etwa 16°, was im Bereich der Werte von 4T@BNNT und 6T@BNNT liegt. Die Messung der zeitaufgelösten Fluoreszenz von freiem und eingekapseltem Nilrot hat ergeben, dass auch in diesem Fall eine Verkürzung der Lebensdauer (von 4091 ps auf 812 ps) erfolgte. Eine solche Verkürzung der Lebensdauer von Chromophoren wird in der Literatur unter anderem mit der Bildung von J-Aggregaten in Zusammenhang gebracht. N2 - This dissertation investigated how the encapsulation of organic dye molecules in boron nitride nanotubes (BNNTs) influences the photophysical properties of the fluorophores. The dyes chosen were alpha-quaterthiophene (4T), alpha-sexithiophene (6T), alpha-octithiophene (8T) and Nile red (NR). The BNNTs used have a nominal diameter of \(5 \pm 2\)nm. To characterize the pure dyes and the hybrid systems consisting of dye and nanotube, a laboratory setup was used that enables PL microscopy in addition to absorption and photoluminescence (PL) spectroscopy. In addition, a time-resolved study of PL (time correlated single photon counting, TCSPC) can be implemented in the ensemble and on individual, separated nano-objects (BNNTs filled with dye). In Chapter 5, the free dyes in solution were first characterized. It has been shown that both 4T and NR are soluble in the solvent used, dimethylformamide (DMF), whereas 6T and 8T show lower solubility. The different profiles of the concentration-dependent PL spectra for 4T and 6T in DMF can probably be attributed to this difference in solubility. In addition, extinction coefficients for 4T and NR were determined using concentration-dependent absorption spectra and there was good agreement with the literature. For 6T and 8T, a determination was not possible due to the low solubility, which is why literature values ​​were used or extrapolated (8T). Chapter 6 detailed the characterization of the BNNTs filled with oligothiophenes. The filling was essentially carried out according to a method published by C. Allard and transferred to the additional fluorophores 4T, 8T and NR. For measurements using UV-Vis spectroscopy in solution or dispersion, it has been shown that the absorption maximum for the dye 6T shifts from 407nm (free 6T) to 506nm (6T@BNNT). The reason for this is probably the formation of J-aggregates inside the tubes. The corresponding PL spectra of free 6T and the hybrid system show no significant differences. For concentration-dependent PL spectra of 6T@BNNT (unlike free 6T in DMF), there is no change in the shape of the curves, which can be interpreted as an indication of successful encapsulation. By combining atomic force and PL microscopy, the outer diameters of individual 6T@BNNT objects could be determined and directly related to their photophysical properties. However, when analyzing the polarization of the emission light from 6T@BNNT depending on the outer diameter, no clear correlation between structure and emission characteristics could be seen. This observation can probably be explained by the fact that only the outer diameter of the (some multi-walled) BNNTs can be determined using atomic force microscopy. The crucial size at this point, however, is the inner diameter of the BNNTs, which influences the alignment and thus also the polarization of the dye molecules. A comparison of the average maximum degree of polarization of the respective hybrid systems showed that 4T@BNNT has the lowest value and 6T@BNNT has the highest value. This confirms the assumption that as the molecule length increases, the polarization increases due to the higher template effect of the tube. 8T@BNNT lies between the other two values, which contradicts this assumption. The average tilt angle of the encapsulated dye molecules relative to the tube axis is about 16° for 4T@BNNT and is therefore slightly larger than that of 6T@BNNT. This also shows that shorter molecules have more steric freedom inside the tubes. For 8T@BNNT the angle is approximately 28° and again contradicts the assumption. TCSPC measurements on free oligothiophene dyes and on the hybrid systems showed that the fluorescence lifetime \(\tau\) for 4T and 6T (each in DMF) increases significantly as a result of encapsulation when the hybrid systems are also dispersed in DMF. The determined values ​​for \(\tau\) of the separated nanoobjects for 4T@BNNT and 6T@BNNT were below the corresponding ones in DMF. For 8T or 8T@BNNT, the lifespan of the separated nanoobjects was significantly shorter compared to the free dye in colloidal suspension. A first approach to elucidate the underlying mechanism was to analyze the TCSPC spectra (for 6T in DMF and 6T@BNNT in DMF) with respect to the individual decay channels. The results obtained indicated that different decay channels dominate for free 6T in DMF than for the hybrid system 6T@BNNT (in DMF). Correlating the fluorescence lifetime of 6T@BNNT with the outer diameter of the nanotubes showed no clear relationship. The characterization of Nile red or NR@BNNT (analogous to the oligothiophenes) took place in Chapter 4. Here, too, there was a shift in the PL spectrum of the fluorophore due to the encapsulation in the BNNTs. However, the PL spectrum of the hybrid system (NR@BNNT) is hypsochromically shifted by about 20 nm. Nile red is also known in the literature as a nanoprobe for determining the permittivity of the solvent or the environment. This allowed an estimation of the relative permittivity inside the BNNTs. The determined value of approx. 4 for an isolated NR@BNNT object indicates a relatively non-polar environment inside the tube. For comparison, the value of free NR in DMF is 47, confirming the relatively high polarity of DMF. The value determined for the average maximum polarization was slightly higher than that of the hybrid systems made of oligothiophenes and nanotubes. The deflection of the NR molecules relative to the tube axis resulted in an angle of approximately 16°, which is in the range of the values ​​for 4T@BNNT and 6T@BNNT. The measurement of the time-resolved fluorescence of free and encapsulated Nile Red showed that in this case too there was a shortening of the lifespan (from 4091 ps to 812 ps). In the literature, such a shortening of the lifespan of chromophores is associated, among other things, with the formation of J-aggregates. KW - Fluoreszenzmikroskopie KW - Spektroskopie KW - Oligothiophene KW - Nanoröhre KW - Nanomaterialien KW - Endohedrale Farbstoffe KW - Nanomaterials KW - Endohedral dyes Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-371150 ER - TY - THES A1 - Schuhmair, Leah Sophia T1 - Etablierung eines in-situ-Immunfluoreszenzfärbeverfahrens zur dreidimensionalen Darstellung und Quantifizierung der Immunzellinfiltration in experimentellen Tumoren T1 - Establishment of an in-situ immunofluorescence staining method for three-dimensional imaging and quantification of immune cell infiltration in experimental tumours N2 - Brustkrebs ist die häufigste diagnostizierte Krebserkrankung weltweit. Trotz der vielfältigen Behandlungsmöglichkeiten endet die Diagnose Brustkrebs in vielen Fällen noch immer tödlich. Aus diesem Grund ist die Entwicklung neuer Therapieansätze wichtig. Ein Therapieansatz, der in den letzten zehn Jahren immer mehr an Bedeutung gewonnen hat, ist die Immuntherapie. Allerdings konnte sie bei Brustkrebs noch keine großen Erfolge erzielen. Ursache hierfür ist die geringe Immunzellinfiltration in Brusttumoren. Um Brustkrebs für Immuntherapie empfänglicher zu machen, müssten Immuntherapeutika in Kombination mit Medikamenten angewendet werden, die die Immunzellinfiltration steigern. Um die Wirksamkeit solcher Medikamente in präklinischen Studien zu testen, braucht es eine Methode, mit der man die T-Zellverteilung innerhalb des Tumors darstellen kann. Für umfassendes Verständnis ist dreidimensionale Darstellung der Zellen im Tumor notwendig, da es einen großen Unterschied macht, ob sich die T-Zellen im Tumorstroma oder in unmittelbarer Nähe zu den Tumorzellen befinden. Die starke Fibrotisierung der Extrazellulären Matrix, die typisch für Brusttumoren ist, erschwert nicht nur die Immunzellinfiltration, sondern auch die Diffusion der fluoreszierenden Antikörper ins Gewebe. Im Zuge dieser Arbeit wurde eine Methode entwickelt, um im dreidimensionalen CD4 und CD8-positive T-Zellen in Brusttumoren darzustellen. Dies gelang mittels Immunfluoreszenzfärbung und anschließender dreidimensionaler Aufnahme mithilfe optischer Sektionierung am Lichtblattmikroskop. Erreicht wurde dies durch deutliche Erhöhung der Inkubationszeiten, aggressive Permeabilisierung des Gewebes, Testen unterschiedlicher Antikörper bzw. Antikörperkombinationen und Entfärbung sowie Klärung des Tumorgewebes. Darüber hinaus konnten erste Schritte in der nachträglichen Bearbeitung der Aufnahmen inklusive Rekonstruktion der Zellen gemacht werden. Für die Anwendung des Verfahrens in Studien zur Medikamentenwirksamkeit ist noch weitere Optimierung notwendig. N2 - Breast cancer is the most frequently diagnosed cancer worldwide. Despite the wide range of treatment options available, the diagnosis of breast cancer is still fatal in many cases. For this reason, the development of new therapeutic approaches is important. One therapeutic approach that has become increasingly important in the last ten years is immunotherapy. However has not yet been very successful in breast cancer. The reason for this is the low level of immune cell infiltration in breast tumours. To make breast cancer more receptive to immunotherapy, immunotherapeutics could be used in combination with drugs that increase immune cell infiltration. In order to test the efficacy of such drugs in preclinical studies, a method is needed that can be used to visualise the T cell distribution within the tumour. For a comprehensive understanding, three-dimensional visualisation of the cells in the tumour is necessary, as it makes a big difference whether the T cells are located in the tumour stroma or in close proximity to the tumour cells. The strong fibrotisation of the extracellular matrix, which is typical of breast tumours, not only makes T cell infiltration, but also the diffusion of the fluorescent antibodies into the tissue difficult. In the course of this work, a method was developed to visualise CD4 and CD8-positive T cells in breast tumours in three dimensions. This was achieved by significantly increasing incubation times, aggressive permeabilisation of the tissue, testing different antibodies and antibody combinations and decolourisation as well as clarification of the tumour tissue. In addition, the first steps were taken in the subsequent processing of the images, including reconstruction of the cells. However further optimisation is still required before the method can be used in drug efficacy studies. KW - Immunfluoreszenz KW - Brustkrebs KW - Dreidimensionales Bild KW - T-Lymphozyt KW - Immuntherapie KW - 3D imaging KW - Tumormikroumgebung KW - immun escape mechanism Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370945 ER - TY - THES A1 - Adolf, Jonas Michael T1 - Die Zusammenarbeit zwischen der stationären beziehungsweise teilstationären psychotherapeutischen Behandlung und niedergelassenen Psychotherapeut:innen T1 - The Collaboration between inpatient and semi-inpatient psychotherapeutic treatment and outpatient psychotherapists N2 - Das Ziel der vorliegenden Arbeit war es die aktuelle Versorgungskontinuität in der psychotherapeutischen Versorgung hinsichtlich der Zusammenarbeit des (teil-)stationären und des ambulanten Sektors aus Sicht der niedergelassenen Psychotherapeut:innen zu untersuchen, diese in den wissenschaftlichen Kontext einzuordnen und – falls möglich – erste Möglichkeiten zur Verbesserung der derzeitigen Versorgungskontinuität aufzuzeigen. In Zusammenarbeit mit dem Arbeitsbereich für Medizinische Psychologie und Psychotherapie im Zentrum für psychische Gesundheit des Universitätsklinikums Würzburg wurde hierzu ein Fragebogen entwickelt und acht ausgewählten psychotherapeutischen Fachgesellschaften beziehungsweise Psychotherapeutenkammern mit der Bitte um Weiterleitung an deren Mitglieder zugesandt. In der vorliegenden Studie wurden – neben einer Globalbeurteilung – im Speziellen die Teil-aspekte des Austauschs, der entsprechenden Rahmenbedingungen und die Bereitstellung des poststationären ambulanten Psychotherapieplatzes betrachtet. Die Studienergebnisse bilden den derzeitigen Status Quo der psychotherapeutischen Versorgungslage aus Sicht der niedergelassenen Psychotherapeut:innen ab und weisen im Zuge dessen auf einige Defizite in den untersuchten Teilaspekten hin. Die aufgestellten Nebenfragestellungen zeigen gleichsam aber auch Ansatzunkte für Lösungen auf. Aufgrund der besonderen Relevanz der aufgezeigten Ergebnisse, gilt es – zur Ermöglichung einer adäquaten kontinuierlichen psychotherapeutischen Versorgung – eine weitergehende Betrach-tung der aufgezeigten Defizite vorzunehmen. Für ein umfassendes Bild sind zudem kongruente Folgearbeiten mit dem Augenmerk auf der Sichtweise der (teil-)stationären Behandlungseinrichtungen und der Patient:innen notwendig. Insbesondere vor dem Hintergrund der limitierten Möglichkeiten der vorliegenden Arbeit gilt es große repräsentative und nationale Studien anzustreben. Hierzu wäre die Etablierung zentral verwalteter Register zur Bündelung der bisherigen und zukünftigen Forschungsarbeiten im Bereich der Psychotherapie wünschenswert. Vor allem vor dem Hintergrund zahlreicher Modellprojekte erscheint dies sinnvoll und könnte einen wichtigen Beitrag zur Optimierung der derzeitigen psychotherapeutischen Forschungs- und Versorgungslage beitragen. N2 - The aim of the study was to identify the current continuity of care regarding psychotherapeutic care and the collaboration of the (semi-)inpatient and outpatient field from point of view of the outpatient psychotherapists. Furthermore, the study wants to integrate the findings in the recent scientific context and wants to reveal improvements regarding the continuity of care in the field of psychotherapy. An explorative survey was created in collaboration with the working group of medical psychology and psychotherapy at the centre for mental health of the university hospital of Würzburg. Eight selected psychotherapeutic organisations (psychotherapy chambers and societies) were invited to conduct the survey to their members. Apart from a general assessment the study took a closer look at the framework conditions, the field of exchange and the care of outpatient psychotherapy to patients after (semi-)inpatient care. The study results describe the current situation of the psychotherapy care in Germany from the point of view of the outpatient psychotherapists and demonstrate on the one hand deficits regarding the analysed aspects and – as part of the side questions – on the other hand possible approaches for improvements. To facilitate a better continuity of psychotherapeutic care, it is important to take a closer look at the demonstrated deficits reported by the study. Moreover further studies are necessary treating the point of view of (semi-)inpatient psychotherapists as well as to conduct large national-wide representative studies, especially against the backdrop of the limited possibilities of the present study. For this purpose, it is helpful to establish a central-managed register to collect all studies in the field of psychotherapy. That could contribute to improve the current situation of research and care in the field of psychotherapy, in particular against the backdrop of the numerous pilot projects. KW - Psychotherapie KW - Medizinische Versorgung KW - Versorgungskontinuität KW - Sektoren KW - Versorgung KW - Psychiatrie Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-371098 ER - TY - THES A1 - Weiß, Eva Maria T1 - Einfluss von Makrophagen auf autophagische Vorgänge in Schwann´schen Zellen unter den Bedingungen von Nervenläsion und genetisch bedingter Neuropathie T1 - Influence of macrophages on Schwann cell autophagy under the conditions of nerve lesion and genetic neuropathy N2 - Charcot-Marie-Tooth (CMT) Neuropathien stellen als häufigste erblich bedingte neurologische Erkrankungen eine Gruppe genetisch heterogener, chronisch progredienter peripherer Polyneuropathien dar. Die Lebensqualität der Patienten ist bei fehlender kurativer Therapieoption vor allem durch motorische und sensorische Defizite deutlich eingeschränkt. In verschiedenen Studien konnte die pathophysiologische Relevanz einer sekundären Entzündungsreaktion, insbesondere durch Makrophagen und Lymphozyten vermittelt, in Mausmodellen dreier CMT1 Subtypen (CMT1A, CMT1B, CMT1X) aufgezeigt werden. Auch in Folge einer Läsion peripherer Nerven ist eine akute Entzündungsreaktion von entscheidender Bedeutung, wobei sich bereits Gemeinsamkeiten zwischen der postläsionalen Waller´schen Degeneration (WD) und CMT1 Neuropathien identifizieren ließen. Während die aktive Beteiligung der Autophagie Schwann´scher Zellen (hier kurz SZ Autophagie genannt) an der Myelindegradation im Falle einer WD jedoch vielfach beschrieben wurde, ist Ähnliches in CMT1 Neuropathien bisher nur unzureichend untersucht. Da in einer Studie in Cx32def Mausmodellen der CMT1X Erkrankung auch nach Reduktion endoneuraler Makrophagen anhaltende Demyelinisierung beobachtet werden konnte, sollte das Vorkommen von SZ Autophagie sowie deren mögliche Beeinflussung durch Makrophagen in diesen Myelinmutanten untersucht werden. In der vorliegenden Arbeit wurden sowohl Wildtyp (Wt) Mäuse in ex vivo und in vivo Modellen einer WD als auch Cx32def Myelinmutanten zweier Altersstufen (4 und 12 Monate) mit einem niedermolekularen CSF1-Rezeptor-Inhibitor (CSF1RI) zur Reduktion endoneuraler Makrophagen behandelt, wobei sich vergleichende histochemische bzw. immunhistochemische Analysen peripherer Nerven behandelter und unbehandelter Tiere anschlossen. Im Rahmen der Etablierung immunhistochemischer Methodik zeigte sich hierbei unter den kontrollierten Bedingungen einer ex vivo Ischiasnervenkultur eine vermehrte Aktivierung der SZ Autophagie in behandelten Wt Mäusen. Auch 4 Monate alte behandelte Cx32def Tiere wiesen, verglichen mit unbehandelten Myelinmutanten bzw. Wt Mäusen derselben Altersstufe, eine vermehrte autophagische Aktivität in SZ auf. Diese scheint sich jedoch im weiteren Verlauf der Erkrankung zu reduzieren, da im Falle der 12 Monate alten Cx32def Modelltiere weniger autophagisch aktive SZ Profile bzw. kaum Unterschiede zwischen behandelten und unbehandelten Tieren beobachtet werden konnten. Die Ergebnisse lassen somit eine mögliche aktive Beteiligung von SZ Autophagie insbesondere in der Pathophysiologie der frühen Phase einer CMT1X Erkrankung sowie deren Beeinflussung durch endoneurale Makrophagen vermuten. Dies sollte vornehmlich in der Entwicklung von Therapiestrategien der CMT1X bedacht werden, da sich eine frühe Reduktion pathophysiologisch relevanter endoneuraler Makrophagen somit auch nachteilig auf die Myelinintegrität auswirken könnte. N2 - Charcot-Marie-Tooth (CMT) neuropathies are the most common hereditary neurological diseases and represent a group of genetically heterogeneous, chronically progressive peripheral polyneuropathies. In the absence of curative treatment options, patients' quality of life is significantly impaired, primarily due to motor and sensory deficits. Various studies have demonstrated the pathophysiological relevance of a secondary inflammatory reaction, in particular mediated by macrophages and lymphocytes, in mouse models of three CMT1 subtypes (CMT1A, CMT1B, CMT1X). An acute inflammatory reaction is also of crucial importance following a lesion of peripheral nerves, whereby similarities between postlesional Wallerian degeneration (WD) and CMT1 neuropathies have already been identified. However, while the active involvement of Schwann cell autophagy (here referred to as SC autophagy) in myelin degradation in WD has been widely described, a similar involvement in CMT1 neuropathies has been insufficiently studied. Since in a study in Cx32def mouse models of CMT1X disease persistent demyelination could be observed even after reduction of endoneural macrophages, the occurrence of SC autophagy and its possible influence by macrophages in these myelin mutants should be investigated. In the present study, both wild-type (Wt) mice in ex vivo and in vivo models of WD and Cx32def myelin mutants of two ages (4 and 12 months) were treated with a small molecule CSF1 receptor inhibitor (CSF1RI) to reduce endoneural macrophages, followed by comparative histochemical and immunohistochemical analyses of peripheral nerves of treated and untreated animals, respectively. During the establishment of immunohistochemical methods, an increased activation of SC autophagy was shown in treated Wt mice under the controlled conditions of ex vivo sciatic nerve culture. Even 4-month-old treated Cx32def animals showed increased autophagic activity in SC compared to untreated myelin mutants or Wt mice of the same age. However, this appears to be reduced as the disease progresses, since in the case of the 12-month-old Cx32def model animals fewer autophagically active SC profiles or hardly any differences between treated and untreated animals could be observed. The results thus suggest a possible active involvement of SC autophagy, particularly in the pathophysiology of the early phase of CMT1X disease and its influence by endoneural macrophages. This should primarily be considered in the development of therapeutic strategies for CMT1X, as an early reduction of pathophysiologically relevant endoneural macrophages could therefore also have a detrimental effect on myelin integrity. KW - Schwann-Zelle KW - M-CSF KW - Autophagie KW - Charcot-Marie-Syndrom KW - Makrophage KW - Schwann´sche Zelle KW - Autophagie KW - hereditäre sensomotorische Neuropathie KW - Makrophagen KW - CSF-1 KW - Immunsystem Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369674 ER - TY - THES A1 - Ruppert [geb. Rapp], Elisabeth Marlene T1 - Einfluss von sozialem Stress und 5-Htt-Genotyp: Quantitative Untersuchung der Morphologie von Neuronen der lateralen Amygdala und der CA3-Region des Hippocampus von Mäusen der Serotonintransporter-Knockout-Linie T1 - Influence of social stress and 5-Htt genotype: Quantitative investigation of the morphology of neurons of the lateral amygdala and the CA3 region of the hippocampus of mice of the serotonin transporter knockout line N2 - In dieser Arbeit wurde der Einfluss sozialer Stresserfahrung sowie des 5-Htt-Genotyps auf die neuronale Morphologie bestimmter Hirnregionen anhand eines Mausmodells untersucht. Es wurde in mit Golgi-Cox gefärbten Gehirnen der 5-HTT-KO-Linie in der lateralen Amygdala (LA) die Apikal- und Basaldendriten pyramidenzellähnlicher Neurone und die Apikaldendriten der Pyramidenzellen der Cornu ammonis (CA)3-Region des Hippocampus mithilfe des Neurolucidasystems rekonstruiert und die so gewonnenen Daten anschließend statistisch ausgewertet. Die erzielten Ergebnisse belegen, dass vor allem die Erfahrung von sozialem Verteidigungsstress aber auch der 5-Htt-Genotyp (WT, HET, KO) im Mausmodell signifikanten Einfluss auf die Morphologie der Neurone der LA und der CA3-Region besitzen. Um die in dieser Arbeit mit allen drei 5-Htt-Genotypen erzielten Ergebnisse der LA-Neurone besser mit den Ergebnissen von Nietzer und Bonn (nur WT, KO) vergleichen zu können (Nietzer et al., 2011), wurden die von mir erhobenen Daten nicht nur in einem 3er-Vergleich, sondern auch einem 2er-Vergleich (WT vs. KO) statistisch analysiert. Untersuchungen der LA-Neurone aller drei 5-Htt-Genotypen zeigen, dass sozialer Stress zu einer Zunahme der Komplexität der Dendritenbäume durch längere und auch stärker verzweigte Dendriten vor allem in der Gruppe der WT-Mäuse führt. HET- und KO-Mäuse zeigten keinen entsprechenden Stress-Effekt. Darüber hinaus zeigten sich deutliche Genotypeffekte. Unabhängig vom Stresserleben besitzen HET-Mäuse längere Dendriten als WT-Mäuse sowie eine höhere Spinedichte als WT- und KO-Mäuse. Die Hypothese, die in der Arbeit von Nietzer et al. aufgestellt wurde, dass eine vollständige 5-HTT-Defizienz zu mehr Spines führt, ließ sich hier weder durch den 3er- noch durch den 2er-Vergleich replizieren. Die Pyramidenzellen der CA3-Region, die in dieser Studie zum ersten Mal analysiert wurden, zeigen in Bezug auf die durch den Stress ausgelösten Veränderungen ein im Vergleich zu den LA-Neuronen entgegengesetzten Effekt. Der soziale Stress führt hier zu einer Dendritenatrophie in der WT-Gruppe mit kürzeren und weniger komplexen Dendriten. Außerdem führte er zu einer geringeren Spinedichte bei den HET-Mäusen. Es zeigten sich klare Genotypeffekte, unabhängig von der Stresserfahrung, mit einer reduzierten Spinedichte der KO-Mäuse gegenüber den WT-Mäusen und einer nur in den Kontrollen detektierten, reduzierten Spinedichte der KO-Mäuse im Vergleich zu den WT- und HET-Mäusen. Sowohl in der LA als auch in der CA3-Region lassen sich Kompensationsmechanismen des 5-HTT-Defizits der HET-Tiere vermuten, über die die KO-Tiere nicht verfügen. Die in LA und CA3 gezeigten gegensätzlichen Auswirkungen des sozialen Stresses weisen auf die unterschiedlichen Funktionen dieser beiden Regionen im Furchtkreislauf und/oder bei der Verarbeitung von Stress hin. Darüber hinaus deutet diese Arbeit darauf hin, dass Arbeiten mit ähnlichen Untersuchungsmethoden und sogar gleichem Untersuchungsmaterial unterschiedliche Ergebnisse liefern können. N2 - In this study, the influence of social stress experience and the 5-Htt genotype on the neuronal morphology of certain brain regions was investigated using a mouse model. The apical and basal dendrites of pyramidal cell-like neurons and the apical dendrites of the pyramidal cells of the cornu ammonis (CA)3 region of the hippocampus were reconstructed in Golgi-Cox-stained brains of the 5-HTT-KO line in the lateral amygdala (LA) using the neurolucida system and the data obtained was then statistically analyzed. The results obtained show that especially the experience of social defense stress but also the 5-Htt genotype (WT, HET, KO) have a significant influence on the morphology of the neurons of the LA and the CA3 region in the mouse model. In order to better compare the results of the LA neurons obtained in this study with all three 5-Htt genotypes with the results of Nietzer and Bonn (WT, KO only) (Nietzer et al., 2011), the data collected by me were statistically analyzed not only in a 3-way comparison, but also in a 2-way comparison (WT vs. KO). Investigations of the LA neurons of all three 5-Htt genotypes show that social stress leads to an increase in the complexity of the dendrite trees due to longer and also more branched dendrites, especially in the group of WT mice. HET and KO mice showed no corresponding stress effect. In addition, there were clear genotype effects. Regardless of the stress experience, HET mice have longer dendrites than WT mice and a higher spin density than WT and KO mice. The hypothesis put forward in the work of Nietzer et al. that complete 5-HTT deficiency leads to more spines could not be replicated here by either the 3-way or 2-way comparison. The pyramidal cells of the CA3 region, which were analyzed for the first time in this study, show an opposite effect compared to the LA neurons with regard to the changes triggered by stress. Here, social stress leads to dendrite atrophy in the WT group with shorter and less complex dendrites. It also led to a lower spin density in the HET mice. There were clear genotype effects, independent of the stress experience, with a reduced spin density in the KO mice compared to the WT mice and a reduced spin density in the KO mice compared to the WT and HET mice, which was only detected in the controls. Compensatory mechanisms for the 5-HTT deficit in the HET animals, which the KO animals do not have, can be assumed in both the LA and the CA3 region. The contrasting effects of social stress shown in LA and CA3 indicate the different functions of these two regions in the fear circuit and/or in the processing of stress. Furthermore, this work suggests that studies using similar research methods and even the same research material may yield different results. KW - Serotoninstoffwechsel KW - Hippocampus KW - Stress KW - Corpus amygdaloideum KW - Ammonshorn KW - CA3-Region KW - laterale Amygdala KW - sozialer Stress KW - Serotonintransporter-Knockout-Linie Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369488 ER - TY - THES A1 - Klüpfel, Marina Anna T1 - Lagedarstellung und -Bewertung durch den Einsatz des Windmühlenmodells - Einführung und Nutzung im Rahmen der SARS-CoV-2 Pandemie T1 - Situational report and assessment using the windmill model – implementation and application during the SARS-CoV-2 pandemic N2 - Bei Großschadensereignissen oder Katastrophen arbeiten die Einsatzkräfte verschiedener Organisationen und Krankenhäuser zusammen, um die Schadenslage zu bewältigen. Für die Koordinierung dieser Einsätze benötigen die Führungskräfte ein möglichst genaues Bild der aktuellen Lage. Auch im Rahmen der SARS-CoV-2- Pandemie war eine Übersicht über die Versorgungslage der Krankenhäuser erforderlich, um mögliche lokale Ressourcenengpässe frühzeitig zu erkennen und durch geeignete Maßnahmen zu beheben. Zu diesem Zweck wurde in Bayern im November 2021 das Windmühlen-Modell eingeführt. Basierend auf einer Online-Plattform meldeten die zuständigen Bezirkskoordinierenden der bayerischen Regierungsbezirke täglich die Versorgungslage ihrer Kliniken anhand der Komponenten Personal, Material und Raum. Außerdem gab es die Möglichkeit zur Dokumentation von Patientenverlegungen. Die über die Windmühlen-Onlineplattform gesammelten Lagemeldungen und dokumentierten Verlegungen des Zeitraums von 21. November 2021 bis 20. Februar 2022 wurden in der vorliegenden Arbeit detailliert aufbereitet. Zusätzlich wurden die erfassten Daten statistisch ausgewertet und mit den örtlichen 7-Tage-Inzidenzwerten des SARS-CoV-2-Virus verglichen. Durch das Windmühlen-Modell konnten Unterschiede in der Versorgungslage zwischen den Regierungsbezirken sehr effektiv sichtbar gemacht werden. Insgesamt waren Intensivstationen deutlich stärker belastet als Normalstationen. Die Versorgungsqualität war in Covid-Bereichen stärker beeinträchtigt als auf Stationen ohne Covid-Patienten. Es konnte nachgewiesen werden, dass die Windmühlen-Lagemeldungen nicht allein die regionalen Inzidenzwerte, sondern die tatsächliche Versorgungssituation vor Ort abbilden. Die dokumentierten Interhospitaltransfers erfolgten von Regionen mit hohen Inzidenzwerten und schlechter Ressourcenverfügbarkeit in Bezirke mit weniger kritischer Versorgungslage. Damit konnten aus den Windmühlen-Lagemeldungen auch konkrete Handlungskonsequenzen, wie strategische Patientenverlegungen, abgeleitet werden. Lagemeldungen sind wichtig für die abgestimmte Zusammenarbeit verschiedener Stellen bei der Bewältigung einer Krise. Die etablierten Systeme zur Lageerfassung sind meist quantitativ ausgelegt und nur wenig skalierbar. Die Anwendung in einem neuen Kontext erfordert oft zeitaufwändige Anpassungen. Im Gegensatz dazu bietet das Windmühlen-Modell eine skalierbare, eher qualitativ ausgerichtete Lagedarstellung und ist aufgrund seines unkomplizierten Aufbaus innerhalb kürzester Zeit für eine Nutzung in verschiedensten Schadenslagen adaptierbar. N2 - During large-scale emergencies and disasters, relief units and hospital staff work together to manage the critical situation. Command and control structures need a detailed situational assessment to coordinate relief efforts. During the SARS-CoV-2 pandemic an overview of hospital supplies and resources was vital to detect local shortages early and find appropriate measures to resolve them. For this purpose, the windmill model was implemented in Bavaria, Germany, in November 2021. Based on an online platform, the seven Bavarian districts gave daily updates on their hospitals’ situation regarding staff, supplies and space. Additionally, there was a tool to record patient transfers to different hospitals. In the dissertation at hand the data collected by the windmill-online platform from 21. November 2021 to 20. February 2022 was evaluated, statistically analyzed and compared to the local 7-day-incidence rates of SARS-CoV-2 virus. The windmill model was able to very effectively showcase differences in strain on the hospital care capacities amongst the districts. Overall, the intensive care units were burdened more heavily than standard care units. The quality of hospital care on Covid-wards was impaired more strongly than on non-Covid-wards. This thesis provides evidence for the windmill situation reports not only depicting the local incidence rates but portraying the actual current hospital care capacities in the districts. The documented patient transfers took place from regions with high incidence rates and poor resource availability to districts with a less critical situation. Thus, specific consequences, like strategic patient transfers, were deduced from situational reports in the windmill model. Situational reports are crucial in collaboration for crisis management. Established systems for situational assessment are often based on quantitative analysis and lack scalability. Using those in a different scenario would require time-consuming adaptations. In contrast the windmill model provides a scalable more qualitatively based situational assessment and is, due to its straightforward format, quickly adaptable to use in any future disaster. KW - Katastrophenmedizin KW - Massenanfall von Verletzten oder Erkrankten KW - Notfallmedizin KW - Einsatzleitung KW - Lagedarstellung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369595 ER - TY - THES A1 - Nair, Radhika Karal T1 - Structural and biochemical characterization of USP28 inhibition by small molecule inhibitors T1 - Strukturelle und biochemische Charakterisierung der Hemmung von USP28 durch niedermolekulare Inhibitoren N2 - Ubiquitination is an important post-translational modification that maintains cellular homeostasis by regulating various biological processes. Deubiquitinases (DUBs) are enzymes that reverse the ubiquitination process by catalyzing the removal of ubiquitin from a substrate. Abnormal expression or function of DUBs is often associated with the onset and progression of various diseases, including cancer. Ubiquitin specific proteases (USPs), which constitute the largest family of DUBs in humans, have become the center of interest as potential targets in cancer therapy as many of them display increased activity or are overexpressed in a range of malignant tumors or the tumor microenvironment. Two related members of the USP family, USP28 and USP25, share high sequence identities but play diverse biological roles. USP28 regulates cell proliferation, oncogenesis, DNA damage repair and apoptosis, whereas USP25 is involved in the anti-viral response, innate immunity and ER-associated degradation in addition to carcinogenesis. USP28 and USP25 also exhibit different oligomeric states – while USP28 is a constitutively active dimer, USP25 assumes an auto-inhibited tetrameric structure. The catalytic domains of both USP28 and USP25 comprise the canonical, globular USP-domain but contain an additional, extended insertion site called USP25/28 catalytic domain inserted domain (UCID) that mediates oligomerization of the proteins. Disruption of the USP25 tetramer leads to the formation of an activated dimeric protein. However, it is still not clear what triggers its activation. Due to their role in maintaining and stabilizing numerous oncoproteins, USP28 and USP25 have emerged as interesting candidates for anti-cancer therapy. Recent advances in small-molecular inhibitor development have led to the discovery of relatively potent inhibitors of USP28 and USP25. This thesis focuses on the structural elucidation of USP28 and the biochemical characterization of USP28/USP25, both in complex with representatives of three out of the eight compound classes reported as USP28/USP25-specific inhibitors. The crystal structures of USP28 in complex with the AZ compounds, Vismodegib and FT206 reveal that all three inhibitor classes bind into the same allosteric pocket distant from the catalytic center, located between the palm and the thumb subdomains (the S1-site). Intriguingly, this binding pocket is identical to the UCID-tip binding interface in the USP25 tetramer, rendering the protein in a locked, inactive conformation. Formation of the binding pocket in USP28 requires a shift in the helix α5, which induces conformational changes and local distortion of the binding channel that typically accommodates the C-terminal tail of Ubiquitin, thus preventing catalysis and abrogating USP28 activity. The key residues of the USP28-inhibitor binding pocket are highly conserved in USP25. Mutagenesis studies of these residues accompanied by biochemical and biophysical assays confirm the proposed mechanism of inhibition and similar binding to USP25. This work provides valuable insights into the inhibition mechanism of the small molecule compounds specifically for the DUBs USP28 and USP25. The USP28-inhibitor complex structures offer a framework to develop more specific and potent inhibitors. N2 - Ubiquitinierung ist eine wichtige posttranslationale Modifikation, die die zelluläre Homöostase aufrechterhält, indem sie verschiedene biologische Prozesse reguliert. Deubiquitinasen (DUBs) sind Enzyme, die den Ubiquitinierungsprozess umkehren, indem sie die Entfernung von Ubiquitin von einem Substrat katalysieren. Eine abnorme Expression oder Funktion von DUBs wird häufig mit dem Auftreten und Fortschreiten verschiedener Krankheiten, einschließlich Krebs, in Verbindung gebracht. Ubiquitin-spezifische Proteasen (USPs), die im Menschen die größte Familie der DUBs bilden, sind als potenzielle Ziele in der Krebstherapie von besonderem Interesse, da viele von ihnen in bösartigen Tumoren oder deren Mikroumgebung abnormal aktiv oder überexprimiert sind. Die zwei eng verwandten Mitglieder der USP-Familie, USP28 und USP25, weisen eine hohe Sequenzidentität auf, sind aber an unterschiedlichen biologischen Prozessen beteiligt. USP28 reguliert die Zellproliferation, die Onkogenese, die Reparatur von DNA-Schäden und die Apoptose, während USP25 eine Rolle bei der antiviralen Reaktion, der angeborenen Immunität, dem ER-assoziierten Abbau und der Carcinogenese spielt. USP28 und USP25 weisen auch unterschiedliche oligomere Zustände auf. Während USP28 ein konstitutiv aktives Dimer bildet, tritt USP25 als auto-inhibiertes Tetramer auf. Strukturell bestehen die katalytischen Domänen sowohl von USP28 als auch von USP25 aus der kanonischen globulären USP-Domäne enthalten jedoch eine zusätzliche Insertion, die als „USP25/28 catalytic domain inserted domain (UCID)“ bezeichnet wird und die Oligomerisierung der Proteine vermittelt. Die Dissoziation des USP25 Tetramers in Dimere führt zu einem aktivierten USP25-Protein. Es ist jedoch immer noch nicht klar, was seine Aktivierung auslöst. Aufgrund ihrer Rolle bei der Aufrechterhaltung und Stabilisierung zahlreicher Onkoproteine haben sich USP28 und USP25 als interessante Kandidaten für die Entwicklung von Medikamenten in der Krebstherapie erwiesen. Jüngste Fortschritte in der Entwicklung von niedermolekularen Inhibitoren haben zur Entdeckung von relativ potenten Inhibitoren von USP28 und USP25 geführt. Diese Arbeit konzentriert sich auf die Strukturaufklärung von USP28 und die biochemische Charakterisierung von USP28/USP25, beide im Komplex mit Vertretern von drei der acht Verbindungsklassen, die als USP28/USP25-spezifische Inhibitoren bekannt sind. Die Kristallstrukturen von USP28 im Komplex mit den AZ-Verbindungen, Vismodegib und FT206 zeigen, dass alle Inhibitoren in einer ähnlichen Region an USP28 binden - einer allosterischen Tasche, die in der Nähe des katalytischen Zentrums liegt und sich zwischen der Handflächen- und der Daumen-Subdomäne befindet. Diese Bindungstasche ist identisch mit der Position, an der der „UCID-tip“ im USP25-Tetramer bindet und das Protein in eine verschränkte, inaktive Konformation versetzt. Die Bildung der Bindungstasche in USP28 erfordert eine Verschiebung der α5-Helix, die zu Konformationsänderungen und einer lokalen Verzerrung des Bindungskanalsführt, der normalerweise den C-terminus des Ubiquitin-Moleküls bindet und so die Katalyse verhindert und die Aktivität von USP28 hemmt. Die Schlüsselreste der USP28-Inhibitor-Bindungstasche sind in USP25 hoch konserviert. Mutagenese-Studien dieser Aminosäuren, begleitet von biochemischen und biophysikalischen Analysen, bestätigen den vorgeschlagenen Mechanismus der Hemmung und eine ähnliche Bindung der Inhibitoren an USP25. Diese Arbeit liefert wertvolle Einblicke in den Hemmungsmechanismus der Kleinmolekülverbindungen, die spezifisch für die DUBs USP28 und USP25 entwickelt worden sind. Die Strukturen der USP28-Inhibitor-Komplexe bieten eine Grundlage für die zukünftige Entwicklung spezifischerer und wirksamerer Inhibitoren. KW - USP KW - Inhibition KW - enzyme KW - crystallography KW - Unique Selling Proposition KW - Inhibition KW - Enzym KW - Kristallographie Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-281742 ER - TY - JOUR A1 - Kampf, Thomas A1 - Bauer, Wolfgang Rudolf A1 - Reiter, Theresa T1 - Improved post-processing strategy for MOLLI based tissue characterization allows application in patients with dyspnoe and impaired left ventricular function JF - Zeitschrift für Medizinische Physik N2 - Contrast and non-contrast MRI based characterization of myocardium by T1-mapping will be of paramount importance to obtain biomarkers, e.g. fibrosis, which determines the risk of heart failure patients. T1-mapping by the standard post-processing of the modified look-locker inversion recovery (MOLLI) lacks of accuracy when trying to reduce its duration, which on the other hand, is highly desirable in patients with heart failure. The recently suggested inversion group fitting (IGF) technique, which considers more parameters for fitting, has a superior accuracy for long T1 times despite a shorter duration. However, for short T1 values, the standard method has a superior precision. A conditional fitting routine is proposed which ideally takes advantage of both algorithms. Materials and methods All measurements were performed on a 1.5 T clinical scanner (ACHIEVA, Philips Healthcare, The Netherlands) using a MOLLI 5(n)3(n)3 prototype with n(heart beats) being a variable waiting time between inversion experiments. Phantom experiments covered a broad range of T1 times, waiting times and heart rates. A saturation recovery experiment served as a gold standard for T1 measurement. All data were analyzed with the standard MOLLI, the IGF fit and the conditional fitting routine and the obtained T1 values were compared with the gold standard. In vivo measurements were performed in a healthy volunteer and a total of 34 patients with normal findings, dilative cardiomyopathy and amyloidosis. Results Theoretical analysis and phantom experiments provided a threshold value for an apparent IGF determining processing with IGF post processing for values above, or switching to the standard technique for values below. This was validated in phantoms and patients measurements. A reduction of the waiting time to 1 instead of 3 heart beats between the inversion experiments showed reliable results. The acquisition time was reduced from 17 to 13 heart beats. The in vivo measurements showed ECV values between 25% (18–33%; SD 0.03) in the healthy, 30% (22–40%; SD 0.04) in patients with DCM and 45% (30–60%; SD 0.9) in patients with amyloidosis. Conclusion The adopted post-processing algorithm determines long T1 values with high accuracy and short T1 values while maintaining a high precision. Based on reduction of waiting time, and independence of heart rate, it shortens breath hold duration and allows fast T1-mapping, which is frequently a prerequisite in patients with cardiac diseases. KW - T1-mapping KW - ECV KW - MOLLI KW - post-processing Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325481 VL - 28 ER - TY - JOUR A1 - Nanadikar, Maithily S. A1 - Vergel Leon, Ana M. A1 - Borowik, Sergej A1 - Hillemann, Annette A1 - Zieseniss, Anke A1 - Belousov, Vsevolod V. A1 - Bogeski, Ivan A1 - Rehling, Peter A1 - Dudek, Jan A1 - Katschinski, Dörthe M. T1 - O2 affects mitochondrial functionality ex vivo JF - Redox Biology N2 - Mitochondria have originated in eukaryotic cells by endosymbiosis of a specialized prokaryote approximately 2 billion years ago. They are essential for normal cell function by providing energy through their role in oxidizing carbon substrates. Glutathione (GSH) is a major thiol-disulfide redox buffer of the cell including the mitochondrial matrix and intermembrane space. We have generated cardiomyocyte-specific Grx1-roGFP2 GSH redox potential (EGSH) biosensor mice in the past, in which the sensor is targeted to the mitochondrial matrix. Using this mouse model a distinct EGSH of the mitochondrial matrix (−278.9 ± 0.4 mV) in isolated cardiomyocytes is observed. When analyzing the EGSH in isolated mitochondria from the transgenic hearts, however, the EGSH in the mitochondrial matrix is significantly oxidized (−247.7 ± 8.7 mV). This is prevented by adding N-Ethylmaleimide during the mitochondria isolation procedure, which precludes disulfide bond formation. A similar reducing effect is observed by isolating mitochondria in hypoxic (0.1–3% O2) conditions that mimics mitochondrial pO2 levels in cellulo. The reduced EGSH is accompanied by lower ROS production, reduced complex III activity but increased ATP levels produced at baseline and after stimulation with succinate/ADP. Altogether, we demonstrate that oxygenation is an essential factor that needs to be considered when analyzing mitochondrial function ex vivo. KW - glutathione redox potential KW - hypoxia KW - mitochondrial matrix KW - Grx1-roGFP Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232217 VL - 22 ER - TY - JOUR A1 - Üçeyler, Nurcan A1 - Urlaub, Daniela A1 - Mayer, Christine A1 - Uehlein, Sabrina A1 - Held, Melissa A1 - Sommer, Claudia T1 - Tumor necrosis factor-α links heat and inflammation with Fabry pain JF - Molecular Genetics and Metabolism N2 - Fabry disease (FD) is an X-linked lysosomal storage disorder associated with pain triggered by heat or febrile infections. We modelled this condition by measuring the cytokine expression of peripheral blood mononuclear cells (PBMC) from FD patients in vitro upon stimulation with heat and lipopolysaccharide (LPS). We enrolled 67 FD patients and 37 healthy controls. We isolated PBMC, assessed their gene expression of selected pro- and anti-inflammatory cytokines, incubated them with heat, LPS, globotriaosylceramide (Gb3), and tumor necrosis factor-α (TNF), and measured TNF secretion in the supernatant and intracellular Gb3 accumulation, respectively. We found increased TNF, interleukin (IL-)1β, and toll-like receptor 4 (TLR4) gene expression in FD men (p < .05 to p < .01). TNF and IL-10 were higher, and IL-4 was lower in the subgroup of FD men with pain compared to controls (p < .05 to p < .01). Hereby, TNF was only increased in FD men with pain and classical mutations (p < .05) compared to those without pain. PBMC from FD patients secreted more TNF upon stimulation with LPS (p < .01) than control PBMC. Incubation with Gb3 and an additional α-galactosidase A inhibitor did not further increase TNF secretion, but incubation with TNF greatly increased the Gb3 load in FD PBMC compared to controls (p < .01). Also, LPS incubation and heat challenge (40 °C) increased Gb3 accumulation in PBMC of patients compared to baseline (p < .05 each), while no alterations were observed in control PBMC. Our data show that TNF holds a crucial role in the pathophysiology of FD associated pain, which may open a novel perspective for analgesic treatment in FD pain. KW - Fabry disease KW - Fabry pain KW - tumor necrosis factor-α KW - peripheral blood mononuclear cells Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229190 VL - 127 ER - TY - JOUR A1 - Kampf, Thomas A1 - Reiter, Theresa A1 - Bauer, Wolfgang Rudolf T1 - An analytical model which determines the apparent T1 for Modified Look-Locker Inversion Recovery – Analysis of the longitudinal relaxation under the influence of discontinuous balanced (classical MOLLI) and spoiled gradient echo readouts JF - Zeitschrift für Medizinische Physik N2 - Quantitative nuclear magnetic resonance imaging (MRI) shifts more and more into the focus of clinical research. Especially determination of relaxation times without/and with contrast agents becomes the foundation of tissue characterization, e.g. in cardiac MRI for myocardial fibrosis. Techniques which assess longitudinal relaxation times rely on repetitive application of readout modules, which are interrupted by free relaxation periods, e.g. the Modified Look-Locker Inversion Recovery = MOLLI sequence. These discontinuous sequences reveal an apparent relaxation time, and, by techniques extrapolated from continuous readout sequences, a putative real T1 is determined. What is missing is a rigorous analysis of the dependence of the apparent relaxation time on its real partner, readout sequence parameters and biological parameters as heart rate. This is provided in this paper for the discontinuous balanced steady state free precession (bSSFP) and spoiled gradient echo readouts. It turns out that the apparent longitudinal relaxation rate is the time average of the relaxation rates during the readout module, and free relaxation period. Knowing the heart rate our results vice versa allow to determine the real T1 from its measured apparent partner. T2 - Ein analytisches Modell, das die apparente T1 Zeit für Modfied Look-Locker Inversion Recovery bestimmt-Analyse der longitudinalen Relaxation unter dem Einfluss diskontinuierlicher balanced (klassische MOLLI) und spoiled gradient echo readouts KW - longitudinal relaxation KW - T1 KW - T2 KW - Lock Locker KW - MOLLI KW - balanced steady state free precession KW - spoiled gradient echo Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325498 VL - 28 ER - TY - JOUR A1 - Minden, Vanessa A1 - Schnetger, Bernhard A1 - Pufal, Gesine A1 - Leonhardt, Sara D. T1 - Antibiotic-induced effects on scaling relationships and on plant element contents in herbs and grasses JF - Ecology and Evolution N2 - Plant performance is correlated with element concentrations in plant tissue, which may be impacted by adverse chemical soil conditions. Antibiotics of veterinary origin can adversely affect plant performance. They are released to agricultural fields via grazing animals or manure, taken up by plants and may be stored, transformed or sequestered by plant metabolic processes. We studied the potential effects of three antibiotics (penicillin, sulfadiazine, and tetracycline) on plant element contents (macro- and microelements). Plant species included two herb species (Brassica napus and Capsella bursa-pastoris) and two grass species (Triticum aestivum and Apera spica-venti), representing two crop species and two noncrop species commonly found in field margins, respectively. Antibiotic concentrations were chosen as to reflect in vivo situations, that is, relatively low concentrations similar to those detected in soils. In a greenhouse experiment, plants were raised in soil spiked with antibiotics. After harvest, macro- and microelements in plant leaves, stems, and roots were determined (mg/g). Results indicate that antibiotics can affect element contents in plants. Penicillin exerted the greatest effect both on element contents and on scaling relationships of elements between plant organs. Roots responded strongest to antibiotics compared to stems and leaves. We conclude that antibiotics in the soil, even in low concentrations, lead to low-element homeostasis, altering the scaling relationships between roots and other plant organs, which may affect metabolic processes and ultimately the performance of a plant. KW - antibiotics KW - homeostasis KW - scaling relationships KW - standardized major axis regression KW - tissue nutrient contents Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224094 VL - 8 ER - TY - JOUR A1 - Duell, Johannes A1 - Lammers, Philip E. A1 - Djuretic, Ivana A1 - Chunyk, Allison G. A1 - Alekar, Shilpa A1 - Jacobs, Ira A1 - Gill, Saar T1 - Bispecific Antibodies in the Treatment of Hematologic Malignancies JF - Clinical Pharmacology & Therapeutics N2 - Monoclonal antibody therapies are an important approach for the treatment of hematologic malignancies, but typically show low single-agent activity. Bispecific antibodies, however, redirect immune cells to the tumor for subsequent lysis, and preclinical and accruing clinical data support single-agent efficacy of these agents in hematologic malignancies, presaging an exciting era in the development of novel bispecific formats. This review discusses recent developments in this area, highlighting the challenges in delivering effective immunotherapies for patients. Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226392 VL - 106 ER - TY - JOUR A1 - Stein, Katharina A1 - Stenchly, Kathrin A1 - Coulibaly, Drissa A1 - Pauly, Alain A1 - Dimobe, Kangbeni A1 - Steffan-Dewenter, Ingolf A1 - Konaté, Souleymane A1 - Goetze, Dethardt A1 - Porembski, Stefan A1 - Linsenmair, K. Eduard T1 - Impact of human disturbance on bee pollinator communities in savanna and agricultural sites in Burkina Faso, West Africa JF - Ecology and Evolution N2 - All over the world, pollinators are threatened by land-use change involving degradation of seminatural habitats or conversion into agricultural land. Such disturbance often leads to lowered pollinator abundance and/or diversity, which might reduce crop yield in adjacent agricultural areas. For West Africa, changes in bee communities across disturbance gradients from savanna to agricultural land are mainly unknown. In this study, we monitored for the impact of human disturbance on bee communities in savanna and crop fields. We chose three savanna areas of varying disturbance intensity (low, medium, and high) in the South Sudanian zone of Burkina Faso, based on land-use/land cover data via Landsat images, and selected nearby cotton and sesame fields. During 21 months covering two rainy and two dry seasons in 2014 and 2015, we captured bees using pan traps. Spatial and temporal patterns of bee species abundance, richness, evenness and community structure were assessed. In total, 35,469 bee specimens were caught on 12 savanna sites and 22 fields, comprising 97 species of 32 genera. Bee abundance was highest at intermediate disturbance in the rainy season. Species richness and evenness did not differ significantly. Bee communities at medium and highly disturbed savanna sites comprised only subsets of those at low disturbed sites. An across-habitat spillover of bees (mostly abundant social bee species) from savanna into crop fields was observed during the rainy season when crops are mass-flowering, whereas most savanna plants are not in bloom. Despite disturbance intensification, our findings suggest that wild bee communities can persist in anthropogenic landscapes and that some species even benefitted disproportionally. West African areas of crop production such as for cotton and sesame may serve as important food resources for bee species in times when resources in the savanna are scarce and receive at the same time considerable pollination service. KW - bee communities KW - cotton KW - sesame KW - species spillover KW - sub-Saharan Africa Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239999 VL - 8 ER - TY - JOUR A1 - Li, Ru-Jin A1 - Han, Muxin A1 - Tessarolo, Jacopo A1 - Holstein, Julian J. A1 - Lübben, Jens A1 - Dittrich, Birger A1 - Volkmann, Christian A1 - Finze, Maik A1 - Jenne, Carsten A1 - Clever, Guido H. T1 - Successive Photoswitching and Derivatization Effects in Photochromic Dithienylethene-Based Coordination Cages JF - ChemPhotoChem N2 - A new series of [Pd2(L)4] cages based on photochromic dithienylethene (DTE) ligands allowed us to gain insight into the successive photoswitching of multiple DTE moieties in a confined metallo-supramolecular assembly. Three new X-ray structures of [Pd2(o-L4)4], [Pd2(o-L1)2(c-L1)2] and [Pd2(c-L1)4] (o-L and c-L = open and closed forms of DTE ligands, respectively) were obtained. The structures deliver snapshots of three different combinations of DTE photoisomeric states within the cage, facilitating a comparison of the all-open with the all-closed, and most notably, an intermediate form where open and closed switches co-exist in the same cage. Moreover, a series of spherical anionic borate clusters was introduced in order to study their roles in the light-controllable host–guest chemistry. The binding guests show higher affinities with the flexible open cage [Pd2(o-L1)4] than with the rigid closed cage [Pd2(c-L1)4]. For the [B12F12]2− guest, thermodynamic data obtained from NMR experiments was compared to results from isothermal titration calorimetry (ITC). KW - coordination cages KW - dithienylethene KW - host-guest systems KW - photoswitches KW - supramolecular chemistry Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-236815 VL - 3 ER - TY - JOUR A1 - Angay, Oguzhan A1 - Friedrich, Mike A1 - Pinnecker, Jürgen A1 - Hintzsche, Henning A1 - Stopper, Helga A1 - Hempel, Klaus A1 - Heinze, Katrin G. T1 - Image-based modeling and scoring of Howell–Jolly Bodies in human erythrocytes JF - Cytometry Part A N2 - The spleen selectively removes cells with intracellular inclusions, for example, detached nuclear fragments in circulating erythrocytes, called Howell–Jolly Bodies (HJBs). With absent or deficient splenic function HJBs appear in the peripheral blood and can be used as a simple and non-invasive risk-indicator for fulminant potentially life-threatening infection after spleenectomy. However, it is still under debate whether counting of the rare HJBs is a reliable measure of splenic function. Investigating HJBs in premature erythrocytes from patients during radioiodine therapy gives about 10 thousand times higher HJB counts than in blood smears. However, we show that there is still the risk of false-positive results by unspecific nuclear remnants in the prepared samples that do not originate from HJBs, but from cell debris residing above or below the cell. Therefore, we present a method to improve accuracy of image-based tests that can be performed even in non-specialized medical institutions. We show how to selectively label HJB-like clusters in human blood samples and how to only count those that are undoubtedly inside the cell. We found a “critical distance” dcrit referring to a relative HJB-Cell distance that true HJBs do not exceed. To rule out false-positive counts we present a simple inside-outside-rule based on dcrit—a robust threshold that can be easily assessed by combining conventional 2D imaging and straight-forward image analysis. Besides data based on fluorescence imaging, simulations of randomly distributed HJB-like objects on realistically modelled cell objects demonstrate the risk and impact of biased counting in conventional analysis. © 2017 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC. KW - fluorescence imaging KW - splenic function KW - Jolly bodies KW - image analysis Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221140 VL - 93 ER - TY - JOUR A1 - Müntze, Jonas A1 - Nordbeck, Peter T1 - Response to “Oral Chaperone Therapy Migalastat for the Treatment of Fabry Disease: Potentials and Pitfalls of Real-World Data” JF - Clinical Pharmacology & Therapeutics N2 - No abstract available Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231600 VL - 106 ER -