TY  - JOUR
A1  - Kraus, Amelie J.
A1  - Brink, Benedikt G.
A1  - Siegel, T. Nicolai
T1  - Efficient and specific oligo-based depletion of rRNA
JF  - Scientific Reports
N2  - In most organisms, ribosomal RNA (rRNA) contributes to >85% of total RNA. Thus, to obtain useful information from RNA-sequencing (RNA-seq) analyses at reasonable sequencing depth, typically, mature polyadenylated transcripts are enriched or rRNA molecules are depleted. Targeted depletion of rRNA is particularly useful when studying transcripts lacking a poly(A) tail, such as some non-coding RNAs (ncRNAs), most bacterial RNAs and partially degraded or immature transcripts. While several commercially available kits allow effective rRNA depletion, their efficiency relies on a high degree of sequence homology between oligonucleotide probes and the target RNA. This restricts the use of such kits to a limited number of organisms with conserved rRNA sequences. In this study we describe the use of biotinylated oligos and streptavidin-coated paramagnetic beads for the efficient and specific depletion of trypanosomal rRNA. Our approach reduces the levels of the most abundant rRNA transcripts to less than 5% with minimal off-target effects. By adjusting the sequence of the oligonucleotide probes, our approach can be used to deplete rRNAs or other abundant transcripts independent of species. Thus, our protocol provides a useful alternative for rRNA removal where enrichment of polyadenylated transcripts is not an option and commercial kits for rRNA are not available.
KW  - parasite biology
KW  - RNA sequencing
KW  - transcriptomics
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224829
VL  - 9
ER  - 
TY  - JOUR
A1  - Kotz, Frederik
A1  - Risch, Patrick
A1  - Arnold, Karl
A1  - Sevim, Semih
A1  - Puigmartí-Luis, Josep
A1  - Quick, Alexander
A1  - Thiel, Michael
A1  - Hrynevich, Andrei
A1  - Dalton, Paul D.
A1  - Helmer, Dorothea
A1  - Rapp, Bastian E.
T1  - Fabrication of arbitrary three-dimensional suspended hollow microstructures in transparent fused silica glass
JF  - Nature Communications
N2  - Fused silica glass is the preferred material for applications which require long-term chemical and mechanical stability as well as excellent optical properties. The manufacturing of complex hollow microstructures within transparent fused silica glass is of particular interest for, among others, the miniaturization of chemical synthesis towards more versatile, configurable and environmentally friendly flow-through chemistry as well as high-quality optical waveguides or capillaries. However, microstructuring of such complex three-dimensional structures in glass has proven evasive due to its high thermal and chemical stability as well as mechanical hardness. Here we present an approach for the generation of hollow microstructures in fused silica glass with high precision and freedom of three-dimensional designs. The process combines the concept of sacrificial template replication with a room-temperature molding process for fused silica glass. The fabricated glass chips are versatile tools for, among other, the advance of miniaturization in chemical synthesis on chip.
KW  - chemical engineering
KW  - fluidics
KW  - materials for optics
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224787
VL  - 10
ER  - 
TY  - JOUR
A1  - Hommers, L. G.
A1  - Richter, J.
A1  - Yang, Y.
A1  - Raab, A.
A1  - Baumann, C.
A1  - Lang, K.
A1  - Schiele, M. A.
A1  - Weber, H.
A1  - Wittmann, A.
A1  - Wolf, C.
A1  - Alpers, G. W.
A1  - Arolt, V.
A1  - Domschke, K.
A1  - Fehm, L.
A1  - Fydrich, T.
A1  - Gerlach, A.
A1  - Gloster, A. T.
A1  - Hamm, A. O.
A1  - Helbig-Lang, S.
A1  - Kircher, T.
A1  - Lang, T.
A1  - Pané-Farré, C. A.
A1  - Pauli, P.
A1  - Pfleiderer, B.
A1  - Reif, A.
A1  - Romanos, M.
A1  - Straube, B.
A1  - Ströhle, A.
A1  - Wittchen, H.-U.
A1  - Frantz, S.
A1  - Ertl, G.
A1  - Lohse, M. J.
A1  - Lueken, U.
A1  - Deckert, J.
T1  - A functional genetic variation of SLC6A2 repressor hsa-miR-579-3p upregulates sympathetic noradrenergic processes of fear and anxiety
JF  - Translational Psychiatry
N2  - Increased sympathetic noradrenergic signaling is crucially involved in fear and anxiety as defensive states. MicroRNAs regulate dynamic gene expression during synaptic plasticity and genetic variation of microRNAs modulating noradrenaline transporter gene (SLC6A2) expression may thus lead to altered central and peripheral processing of fear and anxiety. In silico prediction of microRNA regulation of SLC6A2 was confirmed by luciferase reporter assays and identified hsa-miR-579-3p as a regulating microRNA. The minor (T)-allele of rs2910931 (MAFcases = 0.431, MAFcontrols = 0.368) upstream of MIR579 was associated with panic disorder in patients (pallelic = 0.004, ncases = 506, ncontrols = 506) and with higher trait anxiety in healthy individuals (pASI = 0.029, pACQ = 0.047, n = 3112). Compared to the major (A)-allele, increased promoter activity was observed in luciferase reporter assays in vitro suggesting more effective MIR579 expression and SLC6A2 repression in vivo (p = 0.041). Healthy individuals carrying at least one (T)-allele showed a brain activation pattern suggesting increased defensive responding and sympathetic noradrenergic activation in midbrain and limbic areas during the extinction of conditioned fear. Panic disorder patients carrying two (T)-alleles showed elevated heart rates in an anxiety-provoking behavioral avoidance test (F(2, 270) = 5.47, p = 0.005). Fine-tuning of noradrenaline homeostasis by a MIR579 genetic variation modulated central and peripheral sympathetic noradrenergic activation during fear processing and anxiety. This study opens new perspectives on the role of microRNAs in the etiopathogenesis of anxiety disorders, particularly their cardiovascular symptoms and comorbidities.
KW  - clinical genetics
KW  - psychiatric disorders
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-322497
VL  - 8
ER  - 
TY  - JOUR
A1  - Bury, Susanne
A1  - Soundararajan, Manonmani
A1  - Bharti, Richa
A1  - von Bünau, Rudolf
A1  - Förstner, Konrad U.
A1  - Oelschlaeger, Tobias A.
T1  - The probiotic escherichia coli strain Nissle 1917 combats lambdoid bacteriophages stx and lambda
JF  - Frontiers in Microbiology
N2  - Shiga toxin (Stx) producing E. coli (STEC) such as Enterohemorrhagic E. coli (EHEC) are the major cause of foodborne illness in humans. In vitro studies showed the probiotic Escherichia coil strain Nissle 1917 (EcN) to efficiently inhibit the production of Stx. Life threatening EHEC strains as for example the serotype 0104:H4, responsible for the great outbreak in 2011 in Germany, evolutionary developed from certain E. coll strains which got infected by stx2-encoding lambdoid phages turning the E. coil into lysogenic and subsequently Stx producing strains. Since antibiotics induce stx genes and Stx production, EHEC infected persons are not recommended to be treated with antibiotics. Therefore, EcN might be an alternative medication. However, because even commensal E. coli strains might be converted into Stx-producers after becoming host to a stx encoding prophage, we tested EcN for stx-phage genome integration. Our experiments revealed the resistance of EcN toward not only stx-phages but also against lambda-phages. This resistance was not based on the lack of or by mutated phage receptors. Rather it involved the expression of a phage repressor (pr) gene of a defective prophage in EcN which was able to partially protect E. coli K-12 strain MG1655 against stx and lambda phage infection. Furthermore, we observed EcN to inactivate phages and thereby to protect E. coli K-12 strains against infection by stx- as well as lambda-phages. Inactivation of lambda-phages was due to binding of lambda-phages to LamB of EcN whereas inactivation of stx-phages was caused by a thermostable protein of EcN. These properties together with its ability to inhibit Stx production make EcN a good candidate for the prevention of illness caused by EHEC and probably for the treatment of already infected people.
KW  - probiotic
KW  - E. coli Nissle 1917
KW  - EHEC
KW  - Shiga toxin producing E. coli
KW  - stx-phages
KW  - lambda-phages
KW  - lambdoid prophage
KW  - LamB
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221960
VL  - 9
ER  - 
TY  - JOUR
A1  - Breitinger, Ulrike
A1  - Bahnassawy, Lamiaa M.
A1  - Janzen, Dieter
A1  - Römer, Vera
A1  - Becker, Cord-Michael
A1  - Villmann, Carmen
A1  - Breitinger, Hans-Georg
T1  - PKA and PKC modulators affect ion channel function and internalization of recombinant alpha1 and alpha1-beta glycine receptors
JF  - Frontiers in Molecular Neurosience
N2  - Glycine receptors (GlyRs) are important mediators of fast inhibitory neurotransmission in the mammalian central nervous system. Their function is controlled by multiple cellular mechanisms, including intracellular regulatory processes. Modulation of GlyR function by protein kinases has been reported for many cell types, involving different techniques, and often yielding contradictory results. Here, we studied the effects of protein kinase C (PKC) and cAMP-dependent protein kinase A (PKA) on glycine induced currents in HEK293 cells expressing human homomeric \(\alpha\)1 and heteromeric \(\alpha\)1-\(\beta\) GlyRs using whole-cell patch clamp techniques as well as internalization assays. In whole-cell patch-clamp measurements, modulators were applied in the intracellular buffer at concentrations between 0.1 \(\mu\)M and 0.5 \(\mu\)M. EC50 of glycine increased upon application of the protein kinase activators Forskolin and phorbol-12-myristate-13-acetate (PMA) but decreased in the presence of the PKC inhibitor Staurosporine aglycon and the PKA inhibitor H-89. Desensitization of recombinant \(\alpha\)1 receptors was significantly increased in the presence of Forskolin. Staurosporine aglycon, on the other hand decreased desensitization of heteromeric \(\alpha\)1-\(\beta\) GlyRs. The time course of receptor activation was determined for homomeric \(\alpha\)1 receptors and revealed two simultaneous effects: cells showed a decrease of EC50 after 3-6 min of establishing whole-cell configuration. This effect was independent of protein kinase modulators. All modulators of PKA and PKC, however, produced an additional shift of EC50, which overlay and eventually exceeded the cells intrinsic variation of EC50. The effect of kinase activators was abolished if the corresponding inhibitors were co-applied, consistent with PKA and PKC directly mediating the modulation of GlyR function. Direct effects of PKA-and PKC-modulators on receptor expression on transfected HEK cells were monitored within 15 min of drug application, showing a significant increase of receptor internalization with PKA and PKC activators, while the corresponding inhibitors had no significant effect on receptor surface expression or internalization. Our results confirm the observation that phosphorylation via PKA and PKC has a direct effect on the GlyR ion channel complex and plays an important role in the fine-tuning of glycinergic signaling.
KW  - glycine receptor
KW  - PKA
KW  - PKC
KW  - activators/inhibitors of phosphorylation
KW  - whole-cell currents
KW  - modulation kinetics
KW  - receptor internalization
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220401
VL  - 11
ER  - 
TY  - JOUR
A1  - Bohmann, Ferdinand O.
A1  - Kurka, Natalia
A1  - du Mesnil de Rochemont, Richard
A1  - Gruber, Katharina
A1  - Guenther, Joachim
A1  - Rostek, Peter
A1  - Rai, Heike
A1  - Zickler, Philipp
A1  - Ertl, Michael
A1  - Berlis, Ansgar
A1  - Poli, Sven
A1  - Mengel, Annerose
A1  - Ringleb, Peter
A1  - Nagel, Simon
A1  - Pfaff, Johannes
A1  - Wollenweber, Frank A.
A1  - Kellert, Lars
A1  - Herzberg, Moriz
A1  - Koehler, Luzie
A1  - Haeusler, Karl Georg
A1  - Alegiani, Anna
A1  - Schubert, Charlotte
A1  - Brekenfeld, Caspar
A1  - Doppler, Christopher E. J.
A1  - Onur, Oezguer A.
A1  - Kabbasch, Christoph
A1  - Manser, Tanja
A1  - Pfeilschifter, Waltraud
T1  - Simulation-based training of the rapid evaluation and management of acute stroke (STREAM) — a prospective single-arm multicenter trial
JF  - Frontiers in Neurology
N2  - Introduction: Acute stroke care delivered by interdisciplinary teams is time-sensitive. Simulation-based team training is a promising tool to improve team performance in medical operations. It has the potential to improve process times, team communication, patient safety, and staff satisfaction. We aim to assess whether a multi-level approach consisting of a stringent workflow revision based on peer-to-peer review and 2–3 one-day in situ simulation trainings can improve acute stroke care processing times in high volume neurocenters within a 6 months period.

Methods and Analysis: The trial is being carried out in a pre-test-post-test design at 7 tertiary care university hospital neurocenters in Germany. The intervention is directed at the interdisciplinary multiprofessional stroke teams. Before and after the intervention, process times of all direct-to-center stroke patients receiving IV thrombolysis (IVT) and/or endovascular therapy (EVT) will be recorded. The primary outcome measure will be the “door-to-needle” time of all consecutive stroke patients directly admitted to the neurocenters who receive IVT. Secondary outcome measures will be intervention-related process times of the fraction of patients undergoing EVT and effects on team communication, perceived patient safety, and staff satisfaction via a staff questionnaire.

Interventions: We are applying a multi-level intervention in cooperation with three “STREAM multipliers” from each center. First step is a central meeting of the multipliers at the sponsor's institution with the purposes of algorithm review in a peer-to-peer process that is recorded in a protocol and an introduction to the principles of simulation training and debriefing as well as crew resource management and team communication. Thereafter, the multipliers cooperate with the stroke team trainers from the sponsor's institution to plan and execute 2–3 one-day simulation courses in situ in the emergency department and CT room of the trial centers whereupon they receive teaching materials to perpetuate the trainings.

Clinical Trial Registration: STREAM is a registered trial at https://clinicaltrials.gov/ct2/show/NCT03228251.
KW  - CRM
KW  - thrombolysis (tPA)
KW  - stroke
KW  - emergency care
KW  - simulation training
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369239
SN  - 1664-2295
VL  - 10
ER  - 
TY  - JOUR
A1  - Bolzoni, Francesco
A1  - Esposti, Roberto
A1  - Marchese, Silvia M.
A1  - Pozzi, Nicoló G.
A1  - Ramirez-Pasos, Uri E.
A1  - Isaias, Ioannis U.
A1  - Cavallari, Paolo
T1  - Disrupt of intra-limb APA pattern in parkinsonian patients performing index-finger flexion
JF  - Frontiers in Physiology
N2  - Voluntary movements induce postural perturbations which are counteracted by anticipatory postural adjustments (APAs). These actions are known to build up long fixation chains toward available support points (inter-limb APAs), so as to grant whole body equilibrium. Moreover, recent studies highlighted that APAs also build-up short fixation chains, within the same limb where a distal segment is moved (intra-limb APAs), aimed at stabilizing the proximal segments. The neural structures generating intra-limb APAs still need investigations; the present study aims to compare focal movement kinematics and intra-limb APA latencies and pattern between healthy subjects and parkinsonian patients, assuming the latter as a model of basal ganglia dysfunction. Intra-limb APAs that stabilize the arm when the index-finger is briskly flexed were recorded in 13 parkinsonian patients and in 10 age-matched healthy subjects. Index-finger movement was smaller in parkinsonian patients vs. healthy subjects (p = 0.01) and more delayed with respect to the onset of the prime mover flexor digitorum superficialis (FDS, p < 0.0001). In agreement with the literature, in all healthy subjects the FDS activation was preceded by an inhibitory intra-limb APA in biceps brachii (BB) and anterior deltoid (AD), and almost simultaneous to an excitatory intra-limb APA in triceps brachii (TB). In parkinsonian patients, no significant differences were found for TB and AD intra-limb APA timings, however only four patients showed an inhibitory intra-limb APA in BB, while other four did not show any BB intra-limb APAs and five actually developed a BB excitation. The frequency of occurrence of normal sign, lacking, and inverted BB APAs was different in healthy vs. parkinsonian participants (p = 0.0016). The observed alterations in index-finger kinematics and intra-limb APA pattern in parkinsonian patients suggest that basal ganglia, in addition to shaping the focal movement, may also contribute to intra-limb APA control.
KW  - intra-limb anticipatory postural adjustments
KW  - Parkinson disease
KW  - basal ganglia
KW  - motor control
KW  - human
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369245
SN  - 1664-042X
VL  - 9
ER  - 
TY  - JOUR
A1  - Kessler, Almuth F.
A1  - Frömbling, Greta E.
A1  - Gross, Franziska
A1  - Hahn, Mirja
A1  - Dzokou, Wilfrid
A1  - Ernestus, Ralf-Ingo
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - Effects of tumor treating fields (TTFields) on glioblastoma cells are augmented by mitotic checkpoint inhibition
JF  - Cell Death Discovery
N2  - Tumor treating fields (TTFields) are approved for glioblastoma (GBM) therapy. TTFields disrupt cell division by inhibiting spindle fiber formation. Spindle assembly checkpoint (SAC) inhibition combined with antimitotic drugs synergistically decreases glioma cell growth in cell culture and mice. We hypothesized that SAC inhibition will increase TTFields efficacy. Human GBM cells (U-87 MG, GaMG) were treated with TTFields (200 kHz, 1.7 V/cm) and/or the SAC inhibitor MPS1-IN-3 (IN-3, 4 µM). Cells were counted after 24, 48, and 72 h of treatment and at 24 and 72 h after end of treatment (EOT). Flow cytometry, immunofluorescence microscopy, Annexin-V staining and TUNEL assay were used to detect alterations in cell cycle and apoptosis after 72 h of treatment. The TTFields/IN-3 combination decreased cell proliferation after 72 h compared to either treatment alone (−78.6% vs. TTFields, P = 0.0337; −52.6% vs. IN-3, P = 0.0205), and reduced the number of viable cells (62% less than seeded). There was a significant cell cycle shift from G1 to G2/M phase (P < 0.0001). The apoptotic rate increased to 44% (TTFields 14%, P = 0.0002; IN-3 4%, P < 0.0001). Cell growth recovered 24 h after EOT with TTFields and IN-3 alone, but the combination led to further decrease by 92% at 72 h EOT if IN-3 treatment was continued (P = 0.0288). The combination of TTFields and SAC inhibition led to earlier and prolonged effects that significantly augmented the efficacy of TTFields and highlights a potential new targeted multimodal treatment for GBM.
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325744
VL  - 4
ER  - 
TY  - JOUR
A1  - Kästner, Niklas
A1  - Richter, S. Helene
A1  - Urbanik, Sarah
A1  - Kunert, Joachim
A1  - Waider, Jonas
A1  - Lesch, Klaus-Peter
A1  - Kaiser, Sylvia
A1  - Sachser, Norbert
T1  - Brain serotonin deficiency affects female aggression
JF  - Scientific Reports
N2  - The neurotransmitter serotonin plays a key role in the control of aggressive behaviour. While so far most studies have investigated variation in serotonin levels, a recently created tryptophan hydroxylase 2 (Tph2) knockout mouse model allows studying effects of complete brain serotonin deficiency. First studies revealed increased aggressiveness in homozygous Tph2 knockout mice in the context of a resident-intruder paradigm. Focussing on females, this study aimed to elucidate effects of serotonin deficiency on aggressive and non-aggressive social behaviours not in a test situation but a natural setting. For this purpose, female Tph2 wildtype (n = 40) and homozygous knockout mice (n = 40) were housed with a same-sex conspecific of either the same or the other genotype in large terraria. The main findings were: knockout females displayed untypically high levels of aggressive behaviour even after several days of co-housing. Notably, in response to aggressive knockout partners, they showed increased levels of defensive behaviours. While most studies on aggression in rodents have focussed on males, this study suggests a significant involvement of serotonin also in the control of female aggression. Future research will show, whether the observed behavioural effects are directly caused by the lack of serotonin or by potential compensatory mechanisms.
KW  - animal behaviour
KW  - genetics of the nervous system
KW  - social behaviour
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325386
VL  - 9
ER  - 
TY  - JOUR
A1  - Hecht, Markus
A1  - Meier, Friedegund
A1  - Zimmer, Lisa
A1  - Polat, Bülent
A1  - Loquai, Carmen
A1  - Weishaupt, Carsten
A1  - Forschner, Andrea
A1  - Gutzmer, Ralf
A1  - Utikal, Jochen S.
A1  - Goldinger, Simone M.
A1  - Geier, Michael
A1  - Hassel, Jessica C.
A1  - Balermpas, Panagiotis
A1  - Kiecker, Felix
A1  - Rauschenberg, Ricarda
A1  - Dietrich, Ursula
A1  - Clemens, Patrick
A1  - Berking, Carola
A1  - Grabenbauer, Gerhard
A1  - Schadendorf, Dirk
A1  - Grabbe, Stephan
A1  - Schuler, Gerold
A1  - Fietkau, Rainer
A1  - Distel, Luitpold V.
A1  - Heinzerling, Lucie
T1  - Clinical outcome of concomitant vs interrupted BRAF inhibitor therapy during radiotherapy in melanoma patients
JF  - British Journal of Cancer
N2  - Background:
Concomitant radiation with BRAF inhibitor (BRAFi) therapy may increase radiation-induced side effects but also potentially improve tumour control in melanoma patients.

Methods:
A total of 155 patients with BRAF-mutated melanoma from 17 European skin cancer centres were retrospectively analysed. Out of these, 87 patients received concomitant radiotherapy and BRAFi (59 vemurafenib, 28 dabrafenib), while in 68 patients BRAFi therapy was interrupted during radiation (51 vemurafenib, 17 dabrafenib). Overall survival was calculated from the first radiation (OSRT) and from start of BRAFi therapy (OSBRAFi).

Results:
The median duration of BRAFi treatment interruption prior to radiotherapy was 4 days and lasted for 17 days. Median OSRT and OSBRAFi in the entire cohort were 9.8 and 12.6 months in the interrupted group and 7.3 and 11.5 months in the concomitant group (P=0.075/P=0.217), respectively. Interrupted vemurafenib treatment with a median OSRT and OSBRAFi of 10.1 and 13.1 months, respectively, was superior to concomitant vemurafenib treatment with a median OSRT and OSBRAFi of 6.6 and 10.9 months (P=0.004/P=0.067). Interrupted dabrafenib treatment with a median OSRT and OSBRAFi of 7.7 and 9.8 months, respectively, did not differ from concomitant dabrafenib treatment with a median OSRT and OSBRAFi of 9.9 and 11.6 months (P=0.132/P=0.404). Median local control of the irradiated area did not differ in the interrupted and concomitant BRAFi treatment groups (P=0.619). Skin toxicity of grade ≥2 (CTCAE) was significantly increased in patients with concomitant vemurafenib compared to the group with treatment interruption (P=0.002).

Conclusions:
Interruption of vemurafenib treatment during radiation was associated with better survival and less toxicity compared to concomitant treatment. Due to lower number of patients, the relevance of treatment interruption in dabrafenib treated patients should be further investigated. The results of this analysis indicate that treatment with the BRAFi vemurafenib should be interrupted during radiotherapy. Prospective studies are desperately needed.
KW  - radiation
KW  - radiotherapy
KW  - BRAF
KW  - vemurafenib
KW  - dabrafenib
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227970
VL  - 118
ER  - 
TY  - JOUR
A1  - Knödler, Maren
A1  - Körfer, Justus
A1  - Kunzmann, Volker
A1  - Trojan, Jörg
A1  - Daum, Severin
A1  - Schenk, Michael
A1  - Kullmann, Frank
A1  - Schroll, Sebastian
A1  - Behringer, Dirk
A1  - Stahl, Michael
A1  - Al-Batran, Salah-Eddin
A1  - Hacker, Ulrich
A1  - Ibach, Stefan
A1  - Lindhofer, Horst
A1  - Lordick, Florian
T1  - Randomised phase II trial to investigate catumaxomab (anti-EpCAM × anti-CD3) for treatment of peritoneal carcinomatosis in patients with gastric cancer
JF  - British Journal of Cancer
N2  - Background
Peritoneal carcinomatosis (PC) represents an unfavourable prognostic factor for patients with gastric cancer (GC). Intraperitoneal treatment with the bispecific and trifunctional antibody catumaxomab (EpCAM, CD3), in addition to systemic chemotherapy, could improve elimination of PC.

Methods
This prospective, randomised, phase II study investigated the efficacy of catumaxomab followed by chemotherapy (arm A, 5-fluorouracil, leucovorin, oxaliplatin, docetaxel, FLOT) or FLOT alone (arm B) in patients with GC and PC. Primary endpoint was the rate of macroscopic complete remission (mCR) of PC at the time of second diagnostic laparoscopy/laparotomy prior to optional surgery.

Results
Median follow-up was 52 months. Out of 35 patients screened, 15 were allocated to arm A and 16 to arm B. mCR rate was 27% in arm A and 19% in arm B (p = 0.69). Severe side effects associated with catumaxomab were nausea, infection, abdominal pain, and elevated liver enzymes. Median progression-free (6.7 vs. 5.4 months, p = 0.71) and overall survival (13.2 vs. 13.0 months, p = 0.97) were not significantly different in both treatment arms.

Conclusions
Addition of catumaxomab to systemic chemotherapy was feasible and tolerable in advanced GC. Although the primary endpoint could not be demonstrated, results are promising for future investigations integrating intraperitoneal immunotherapy into a multimodal treatment strategy.
KW  - cancer immunotherapy
KW  - gastric cancer
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325938
VL  - 119
ER  - 
TY  - JOUR
A1  - Altieri, Barbara
A1  - Di Dato, Carla
A1  - Martini, Chiara
A1  - Sciammarella, Concetta
A1  - Di Sarno, Antonella
A1  - Colao, Annamaria
A1  - Faggiano, Antongiulio
T1  - Bone Metastases in Neuroendocrine Neoplasms: From Pathogenesis to Clinical Management
JF  - Cancers
N2  - Bone represents a common site of metastases for several solid tumors. However, the ability of neuroendocrine neoplasms (NENs) to localize to bone has always been considered a rare and late event. Thanks to the improvement of therapeutic options, which results in longer survival, and of imaging techniques, particularly after the introduction of positron emission tomography (PET) with gallium peptides, the diagnosis of bone metastases (BMs) in NENs is increasing. The onset of BMs can be associated with severe skeletal complications that impair the patient's quality of life. Moreover, BMs negatively affect the prognosis of NEN patients, bringing out the lack of curative treatment options for advanced NENs. The current knowledge on BMs in gastro-entero-pancreatic (GEP) and bronchopulmonary (BP) NENs is still scant and is derived from a few retrospective studies and case reports. This review aims to perform a critical analysis of the evidence regarding the role of BMs in GEP- and BP-NENs, focusing on the molecular mechanisms underlining the development of BMs, as well as clinical presentation, diagnosis, and treatment of BMs, in an attempt to provide suggestions that can be used in clinical practice.
KW  - neuroendocrine neoplasms
KW  - bone metastases
KW  - bone microenvironment
KW  - skeletal-related events
KW  - epithelial-to-mesenchymal transition
KW  - microRNA
KW  - prognosis
KW  - treatment
KW  - denosumab
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221079
VL  - 11
ER  - 
TY  - JOUR
A1  - El-Helou, Sabine M.
A1  - Biegner, Anika-Kerstin
A1  - Bode, Sebastian
A1  - Ehl, Stephan R.
A1  - Heeg, Maximilian
A1  - Maccari, Maria E.
A1  - Ritterbusch, Henrike
A1  - Speckmann, Carsten
A1  - Rusch, Stephan
A1  - Scheible, Raphael
A1  - Warnatz, Klaus
A1  - Atschekzei, Faranaz
A1  - Beider, Renata
A1  - Ernst, Diana
A1  - Gerschmann, Stev
A1  - Jablonka, Alexandra
A1  - Mielke, Gudrun
A1  - Schmidt, Reinhold E.
A1  - Schürmann, Gesine
A1  - Sogkas, Georgios
A1  - Baumann, Ulrich H.
A1  - Klemann, Christian
A1  - Viemann, Dorothee
A1  - Bernuth, Horst von
A1  - Krüger, Renate
A1  - Hanitsch, Leif G.
A1  - Scheibenbogen, Carmen M.
A1  - Wittke, Kirsten
A1  - Albert, Michael H.
A1  - Eichinger, Anna
A1  - Hauck, Fabian
A1  - Klein, Christoph
A1  - Rack-Hoch, Anita
A1  - Sollinger, Franz M.
A1  - Avila, Anne
A1  - Borte, Michael
A1  - Borte, Stephan
A1  - Fasshauer, Maria
A1  - Hauenherm, Anja
A1  - Kellner, Nils
A1  - Müller, Anna H.
A1  - Ülzen, Anett
A1  - Bader, Peter
A1  - Bakhtiar, Shahrzad
A1  - Lee, Jae-Yun
A1  - Heß, Ursula
A1  - Schubert, Ralf
A1  - Wölke, Sandra
A1  - Zielen, Stefan
A1  - Ghosh, Sujal
A1  - Laws, Hans-Juergen
A1  - Neubert, Jennifer
A1  - Oommen, Prasad T.
A1  - Hönig, Manfred
A1  - Schulz, Ansgar
A1  - Steinmann, Sandra
A1  - Klaus, Schwarz
A1  - Dückers, Gregor
A1  - Lamers, Beate
A1  - Langemeyer, Vanessa
A1  - Niehues, Tim
A1  - Shai, Sonu
A1  - Graf, Dagmar
A1  - Müglich, Carmen
A1  - Schmalzing, Marc T.
A1  - Schwaneck, Eva C.
A1  - Tony, Hans-Peter
A1  - Dirks, Johannes
A1  - Haase, Gabriele
A1  - Liese, Johannes G.
A1  - Morbach, Henner
A1  - Foell, Dirk
A1  - Hellige, Antje
A1  - Wittkowski, Helmut
A1  - Masjosthusmann, Katja
A1  - Mohr, Michael
A1  - Geberzahn, Linda
A1  - Hedrich, Christian M.
A1  - Müller, Christiane
A1  - Rösen-Wolff, Angela
A1  - Roesler, Joachim
A1  - Zimmermann, Antje
A1  - Behrends, Uta
A1  - Rieber, Nikolaus
A1  - Schauer, Uwe
A1  - Handgretinger, Rupert
A1  - Holzer, Ursula
A1  - Henes, Jörg
A1  - Kanz, Lothar
A1  - Boesecke, Christoph
A1  - Rockstroh, Jürgen K.
A1  - Schwarze-Zander, Carolynne
A1  - Wasmuth, Jan-Christian
A1  - Dilloo, Dagmar
A1  - Hülsmann, Brigitte
A1  - Schönberger, Stefan
A1  - Schreiber, Stefan
A1  - Zeuner, Rainald
A1  - Ankermann, Tobias
A1  - Bismarck, Philipp von
A1  - Huppertz, Hans-Iko
A1  - Kaiser-Labusch, Petra
A1  - Greil, Johann
A1  - Jakoby, Donate
A1  - Kulozik, Andreas E.
A1  - Metzler, Markus
A1  - Naumann-Bartsch, Nora
A1  - Sobik, Bettina
A1  - Graf, Norbert
A1  - Heine, Sabine
A1  - Kobbe, Robin
A1  - Lehmberg, Kai
A1  - Müller, Ingo
A1  - Herrmann, Friedrich
A1  - Horneff, Gerd
A1  - Klein, Ariane
A1  - Peitz, Joachim
A1  - Schmidt, Nadine
A1  - Bielack, Stefan
A1  - Groß-Wieltsch, Ute
A1  - Classen, Carl F.
A1  - Klasen, Jessica
A1  - Deutz, Peter
A1  - Kamitz, Dirk
A1  - Lassy, Lisa
A1  - Tenbrock, Klaus
A1  - Wagner, Norbert
A1  - Bernbeck, Benedikt
A1  - Brummel, Bastian
A1  - Lara-Villacanas, Eusebia
A1  - Münstermann, Esther
A1  - Schneider, Dominik T.
A1  - Tietsch, Nadine
A1  - Westkemper, Marco
A1  - Weiß, Michael
A1  - Kramm, Christof
A1  - Kühnle, Ingrid
A1  - Kullmann, Silke
A1  - Girschick, Hermann
A1  - Specker, Christof
A1  - Vinnemeier-Laubenthal, Elisabeth
A1  - Haenicke, Henriette
A1  - Schulz, Claudia
A1  - Schweigerer, Lothar
A1  - Müller, Thomas G.
A1  - Stiefel, Martina
A1  - Belohradsky, Bernd H.
A1  - Soetedjo, Veronika
A1  - Kindle, Gerhard
A1  - Grimbacher, Bodo
T1  - The German national registry of primary immunodeficiencies (2012-2017)
JF  - Frontiers in Immunology
N2  - Introduction: The German PID-NET registry was founded in 2009, serving as the first national registry of patients with primary immunodeficiencies (PID) in Germany. It is part of the European Society for Immunodeficiencies (ESID) registry. The primary purpose of the registry is to gather data on the epidemiology, diagnostic delay, diagnosis, and treatment of PIDs. 

Methods: Clinical and laboratory data was collected from 2,453 patients from 36 German PID centres in an online registry. Data was analysed with the software Stata® and Excel. 

Results: The minimum prevalence of PID in Germany is 2.72 per 100,000 inhabitants. Among patients aged 1-25, there was a clear predominance of males. The median age of living patients ranged between 7 and 40 years, depending on the respective PID. Predominantly antibody disorders were the most prevalent group with 57% of all 2,453 PID patients (including 728 CVID patients). A gene defect was identified in 36% of patients. Familial cases were observed in 21% of patients. The age of onset for presenting symptoms ranged from birth to late adulthood (range 0-88 years). Presenting symptoms comprised infections (74%) and immune dysregulation (22%). Ninety-three patients were diagnosed without prior clinical symptoms. Regarding the general and clinical diagnostic delay, no PID had undergone a slight decrease within the last decade. However, both, SCID and hyper IgE-syndrome showed a substantial improvement in shortening the time between onset of symptoms and genetic diagnosis. Regarding treatment, 49% of all patients received immunoglobulin G (IgG) substitution (70%-subcutaneous; 29%-intravenous; 1%-unknown). Three-hundred patients underwent at least one hematopoietic stem cell transplantation (HSCT). Five patients had gene therapy. 

Conclusion: The German PID-NET registry is a precious tool for physicians, researchers, the pharmaceutical industry, politicians, and ultimately the patients, for whom the outcomes will eventually lead to a more timely diagnosis and better treatment.
KW  - registry for primary immunodeficiency
KW  - primary immunodeficiency (PID)
KW  - German PID-NET registry
KW  - PID prevalence
KW  - European Society for Immunodeficiencies (ESID)
KW  - IgG substitution therapy
KW  - CVID
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226629
VL  - 10
ER  - 
TY  - JOUR
A1  - Butt, Elke
A1  - Raman, Dayanidhi
T1  - New frontiers for the cytoskeletal protein LASP1
JF  - Frontiers in Oncology
N2  - In the recent two decades, LIM and SH3 protein 1 (LASP1) has been developed from a simple actin-binding structural protein to a tumor biomarker and subsequently to a complex, nuclear transcriptional regulator. Starting with a brief historical perspective, this review will mainly compare and contrast LASP1 and LASP2 from the angle of the newest data and importantly, examine their role in transcriptional regulation. We will summarize the current knowledge through pictorial models and tables including the roles of different microRNAs in the differential regulation of LASP1 levels and patient outcome rather than specify in detail all tumor entities. Finally, the novel functional roles of LASP1 in secretion of vesicles, expression of matrix metalloproteinases and transcriptional regulation as well as the activation of survival and proliferation pathways in different cancer types are described.
KW  - LASP1
KW  - LASP2
KW  - transcriptional regulation
KW  - nuclear role
KW  - matrix metalloproteinases
KW  - AP1
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221975
VL  - 8
ER  - 
TY  - JOUR
A1  - Doppler, Christopher E. J.
A1  - Meyer, Linda
A1  - Dovern, Anna
A1  - Stühmer-Beckh, Jaro
A1  - Weiss, Peter H.
A1  - Fink, Gereon R.
T1  - Differential impact of social and monetary reward on procedural learning and consolidation in aging and its structural correlates
JF  - Frontiers in Aging Neuroscience
N2  - In young (n = 36, mean +/- SD: 24.8 +/- 4.5 years) and older (n = 34, mean +/- SD: 65.1 +/- 6.5 years) healthy participants, we employed a modified version of the Serial Reaction Time task to measure procedural learning (PL) and consolidation while providing monetary and social reward. Using voxel-based morphometry (VBM), we additionally determined the structural correlates of reward-related motor performance (RMP) and PL. Monetary reward had a beneficial effect on PL in the older subjects only. In contrast, social reward significantly enhanced PL in the older and consolidation in the young participants. VBM analyses revealed that motor performance related to monetary reward was associated with larger grey matter volume (GMV) of the left striatum in the young, and motor performance related to social reward with larger GMV of the medial orbitofrontal cortex in the older group. The differential effects of social reward in young (improved consolidation) and both social and monetary rewards in older (enhanced PL) healthy subjects point to the potential of rewards for interventions targeting aging-associated motor decline or stroke-induced motor deficits.
KW  - serial reaction time task
KW  - procedural learning
KW  - reinforcement learning
KW  - voxel-based morphometry
KW  - motor aging
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222394
VL  - 11
ER  - 
TY  - JOUR
A1  - Doppler, Kathrin
A1  - Brockmann, Kathrin
A1  - Sedghi, Annahita
A1  - Wurster, Isabel
A1  - Volkmann, Jens
A1  - Oertel, Wolfgang H.
A1  - Sommer, Claudia
T1  - Dermal phospho-alpha-synuclein deposition in patients with Parkinson's disease and mutation of the glucocerebrosidase gene
JF  - Frontiers in Neurology
N2  - Heterozygous mutations in the glucocerebrosidase gene (GBA1) represent the most common genetic risk factor for Parkinson's disease (PD) and are histopathologically associated with a widespread load of alpha-synuclein in the brain. Therefore, PD patients with GBA1 mutations are a cohort of high interest for clinical trials on disease-modifying therapies targeting alpha-synuclein. There is evidence that detection of phospho-alpha-synuclein (p-syn) in dermal nerve fibers might be a biomarker for the histopathological identification of PD patients even at premotor or very early stages of disease. It is so far unknown whether dermal p-syn deposition can also be found in PD patients with GBA1 mutations and may serve as a biomarker for PD in these patients. Skin biopsies of 10 PD patients with different GBA1 mutations (six N3705, three E326K, one L444P) were analyzed by double-immunofluorescence labeling with anti-p-syn and anti-protein gene product 9.5 (PGP9.5, axonal marker) to detect intraaxonal p-syn deposition. Four biopsy sites (distal, proximal leg, paravertebral Th10, and C7) per patient were studied. P-syn was found in six patients (three N370S, three E326K). P-syn deposition was mainly detected in autonomic nerve fibers, but also in somatosensory fibers and was not restricted to a certain GBA1 mutation. In summary, dermal p-syn in PD patients with GBA1 mutations seems to offer a similar distribution and frequency as observed in patients without a known mutation. Skin biopsy may be suitable to study p-syn deposition in these patients or even to identify premotor patients with GBA1 mutations.
KW  - Parkinson's disease
KW  - glucocerebrosidase mutation
KW  - alpha-synuclein
KW  - skin biopsy
KW  - biomarker
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222828
VL  - 9
ER  - 
TY  - THES
A1  - Kemmer, Luisa Diana
T1  - Darstellung von Inflammation in Atherosklerose mit dem CXCR4-gerichteten PET-Tracer \(^{68}\)Ga-Pentixafor im Vergleich zur \(^{18}\)F-FDG-PET/CT
T1  - Imaging inflammation in atherosclerosis with the CXCR4-targeted PET tracer \(^{68}\)Ga-Pentixafor compared to \(^{18}\)F-FDG-PET/CT
N2  - Herz-Kreislauf-Erkrankungen zählen zu den häufigsten Todesursachen weltweit. Ein ihr zugrundeliegender pathologischer Prozess ist die Atherosklerose. Die Ruptur eines atheroskelrotischen Plaques kann lebensbedrohlich sein. Derzeit existieren weder ein evaluierter Biomarker noch eine Bildgebungstechnik, die das Risiko einer solchen Plaqueruptur und eines nachfolgenden akuten kardiovaskulären Ereignisses vorhersagen können. Um die bildgebenden Verfahren zur Detektion der Atherosklerose zu optimieren, wurde in dieser Dissertationsarbeit untersucht, ob der PET/CT-Tracer 68Ga-Pentixafor im Vergleich zu 18F-FDG bessere Ergebnisse in der Diagnostik der Atherosklerose erzielen kann. 
Insgesamt wurden 144 onkologische Patienten in die Studie einbezogen, bei denen die 18F-FDG-PET/CT sowie 68Ga-Pentifaxor-PET/CT aus klinischen Gründen indiziert waren. Befunde, bei denen eine gegenüber dem Hintergrund vermehrte Speicherung ohne physiologische Erklärung nachgewiesen werden konnte, wurden als positiv bewertet. Um Unterschiede zwischen den Patienten außer Acht lassen zu können, wurde die target-to-background-ratio (TBR) berechnet. Zur Beschreibung der Speicherintensität einer Läsion wurde der standardized uptake value (SUV) bestimmt. 
Nach Auswertung der Daten zeigte sich eine mäßige Korrelation der Anzahl von 68Ga-Pentixafor-positiven Läsionen mit der Anzahl der 18F-FDG positiven Läsionen. Die CXCR4-gerichtete Bildgebung mit 68Ga-Pentixafor identifizierte mehr Läsionen als die 18F-FDG-PET/CT. Bezüglich ihres Verteilungsmusters wiesen die beiden Tracer eine geringe Korrelation auf. Die Aufnahmeintensität beider Tracer korrelierte umgekehrt mit dem Ausmaß der Verkalkung. Stark verkalkte Plaques zeigten die niedrigste Traceraufnahme für beide PET-Tracer. 
Weitere Studien zur Aufklärung der zugrunde liegenden biologischen Mechanismen und Quellen der CXCR4-Positivität sind in hohem Maße gerechtfertigt.
N2  - Cardiovascular diseases are among the most common causes of death worldwide. A pathological process underlying these diseases is atherosclerosis. The rupture of an atherosclerotic plaque can be life-threatening. Currently, there is neither an evaluated biomarker nor an imaging technique that can predict the risk of such a plaque rupture and subsequent acute cardiovascular event. To optimize imaging methods for the detection of atherosclerosis, this dissertation investigated whether the PET/CT tracer 68Ga-Pentixafor can achieve better diagnostic results for atherosclerosis compared to
18F-FDG. A total of 144 oncological patients were included in the study, for whom 18F-FDG-PET/CT and 68Ga-Pentixafor-PET/CT were clinically indicated. Lesions showing increased uptake compared to the background without physiological explanation were rated as positive. To disregard differences between patients, the target-to-background ratio (TBR) was calculated. To describe the uptake intensity of a lesion, the standardized uptake value (SUV) was determined. After evaluating the data, a moderate correlation was observed between the number of 68Ga-Pentixafor-positive lesions and the number of 18F-FDG-positive lesions. CXCR4-targeted imaging with 68Ga-Pentixafor identified more lesions than 18F-FDG-PET/CT. Regarding their distribution patterns, the two tracers showed a low correlation. The uptake intensity of both tracers inversely correlated with the extent of calcification. Highly calcified plaques exhibited the lowest tracer uptake for both PET tracers. Further studies to elucidate the underlying biological mechanisms and sources of CXCR4 positivity are highly warranted.
KW  - Arteriosklerose
KW  - Positronen-Emissions-Tomografie
KW  - Chemokinrezpetor
KW  - CXCR4
KW  - \(^{68}\)Ga-Pentixafor
KW  - \(^{18}\)F-FDG
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360013
ER  - 
TY  - THES
A1  - Zillig, Anna-Lena Christina
T1  - Einfluss von Sicherheit auf die Schmerzverarbeitung
T1  - The influence of safety on pain perception
N2  - Im Rahmen des interdisziplinären Promotionsschwerpunkts Resilienzfaktoren der Schmerzverarbeitung des evangelischen Studienwerks in Zusammenarbeit mit der Julius-Maximilians-Universität Würzburg und der Otto-Friedrich-Universität Bamberg untersuche ich in diesem Promotionsprojekt den Einfluss von Sicherheit auf die Schmerzverarbeitung. Es ist bekannt, dass die Schmerzverarbeitung durch Emotionen moduliert werden kann. Man geht davon aus, dass negative Emotionen den Schmerz in der Regel verstärken, während positive Emotionen zu einer Schmerzreduktion führen. Frühere Studien fanden heraus, dass die Erwartung eines aversiven Ereignisses zu Bedrohung und stärkeren Schmerzen führt. Es stellt sich die Frage, ob das Gegenteil von Bedrohung, nämlich Sicherheit, zu einer Verringerung der Schmerzen führen kann. Um diese Hypothese zu untersuchen, habe ich drei Experimente an gesunden ProbandInnen durchgeführt.
N2  - I am investigating the influence of safety on pain processing in the present dissertation project as part of the interdisciplinary doctoral program on resilience factors of pain processing of the Evangelisches Studienwerk in cooperation with the Julius-Maximilians-University of Würzburg and the Otto-Friedrich-University of Bamberg. It is known that pain processing is susceptible to an individual’s emotional state, such that negative emotions mostly increase pain while positive emotions lead to a pain decrease. Previous studies found that the anticipation of an aversive event induces threat and elevated pain. The question arises whether the exact opposite, namely safety, can lead to a reduction in pain. To investigate this hypothesis, I conducted three experiments in healthy volunteers.
KW  - Sicherheit
KW  - Schmerzverarbeitung
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359282
ER  - 
TY  - JOUR
A1  - Vural, Atay
A1  - Doppler, Kathrin
A1  - Meinl, Edgar
T1  - Autoantibodies Against the Node of Ranvier in Seropositive Chronic Inflammatory Demyelinating Polyneuropathy: Diagnostic, Pathogenic, and Therapeutic Relevance
JF  - Frontiers in Immunology
N2  - Discovery of disease-associated autoantibodies has transformed the clinical management of a variety of neurological disorders. Detection of autoantibodies aids diagnosis and allows patient stratification resulting in treatment optimization. In the last years, a set of autoantibodies against proteins located at the node of Ranvier has been identified in patients with chronic inflammatory demyelinating polyneuropathy (CIDP). These antibodies target neurofascin, contactin1, or contactin-associated protein 1, and we propose to name CIDP patients with these antibodies collectively as seropositive. They have unique clinical characteristics that differ from seronegative CIDP. Moreover, there is compelling evidence that autoantibodies are relevant for the pathogenesis. In this article, we review the current knowledge on the characteristics of autoantibodies against the node of Ranvier proteins and their clinical relevance in CIDP. We start with a description of the structure of the node of Ranvier followed by a summary of assays used to identify seropositive patients; and then, we describe clinical features and characteristics linked to seropositivity. We review knowledge on the role of these autoantibodies for the pathogenesis with relevance for the emerging concept of nodopathy/paranodopathy and summarize the treatment implications.
KW  - autoantibody
KW  - seropositive
KW  - chronic inflammatory demyelinating polyneuropathy
KW  - node of Ranvier
KW  - paranode
KW  - neurofascin
KW  - contactin
KW  - contactin-associated protein 1
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233279
VL  - 9
ER  - 
TY  - JOUR
A1  - Siebert, Claudia
A1  - Ciato, Denis
A1  - Murakami, Masanori
A1  - Frei-Stuber, Ludwig
A1  - Perez-Rivas, Luis Gustavo
A1  - Monteserin-Garcia, José Luis
A1  - Nölting, Svenja
A1  - Maurer, Julian
A1  - Feuchtinger, Annette
A1  - Walch, Axel K.
A1  - Haak, Harm R.
A1  - Bertherat, Jérôme
A1  - Mannelli, Massimo
A1  - Fassnacht, Martin
A1  - Korpershoek, Esther
A1  - Reincke, Martin
A1  - Stalla, Günter K.
A1  - Hantel, Constanze
A1  - Beuschlein, Felix
T1  - Heat Shock Protein 90 as a Prognostic Marker and Therapeutic Target for Adrenocortical Carcinoma
JF  - Frontiers in Endocrinology
N2  - Background: Adrenocortical carcinoma (ACC) is a rare tumor entity with restricted therapeutic opportunities. HSP90 (Heat Shock Protein 90) chaperone activity is fundamental for cell survival and contributes to different oncogenic signaling pathways. Indeed, agents targeting HSP90 function have shown therapeutic efficacy in several cancer types. We have examined the expression of HSP90 in different adrenal tumors and evaluated the use of HSP90 inhibitors in vitro as possible therapy for ACC.

Methods: Immunohistochemical expression of HSP90 isoforms was investigated in different adrenocortical tumors and associated with clinical features. Additionally, a panel of N-terminal (17-allylamino-17-demethoxygeldanamycin (17-AAG), luminespib, and ganetespib) and C-terminal (novobiocin and silibinin) HSP90 inhibitors were tested on various ACC cell lines.

Results: Within adrenocortical tumors, ACC samples exhibited the highest expression of HSP90β. Within a cohort of ACC patients, HSP90β expression levels were inversely correlated with recurrence-free and overall survival. In functional assays, among five different compounds tested luminespib and ganetespib induced a significant decrease in cell viability in single as well as in combined treatments with compounds of the clinically used EDP-M scheme (etoposide, doxorubicin, cisplatin, mitotane). Inhibition of cell viability correlated furthermore with a decrease in proliferation, in cell migration and an increase in apoptosis. Moreover, analysis of cancer pathways indicated a modulation of the ERK1/2—and AKT—pathways by luminespib and ganetespib treatment.

Conclusions: Our findings emphasize HSP90 as a marker with prognostic impact and promising target with N-terminal HSP90 inhibitors as drugs with potential therapeutic efficacy toward ACC.
KW  - adrenal gland
KW  - cortisol
KW  - N-terminal HSP90 inhibitors
KW  - C-terminal HSP90 inhibitors
KW  - prognostic marker
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238029
VL  - 10
ER  - 
TY  - JOUR
A1  - Nguyen, Minh-Thu
A1  - Saising, Jongkon
A1  - Tribelli, Paula Maria
A1  - Nega, Mulugeta
A1  - Diene, Seydina M.
A1  - François, Patrice
A1  - Schrenzel, Jacques
A1  - Spröer, Cathrin
A1  - Bunk, Boyke
A1  - Ebner, Patrick
A1  - Hertlein, Tobias
A1  - Kumari, Nimerta
A1  - Härtner, Thomas
A1  - Wistuba, Dorothee
A1  - Voravuthikunchai, Supayang P.
A1  - Mäder, Ulrike
A1  - Ohlsen, Knut
A1  - Götz, Friedrich
T1  - Inactivation of farR Causes High Rhodomyrtone Resistance and Increased Pathogenicity in Staphylococcus aureus
JF  - Frontiers in Microbiology
N2  - Rhodomyrtone (Rom) is an acylphloroglucinol antibiotic originally isolated from leaves of Rhodomyrtus tomentosa. Rom targets the bacterial membrane and is active against a wide range of Gram-positive bacteria but the exact mode of action remains obscure. Here we isolated and characterized a spontaneous Rom-resistant mutant from the model strain Staphylococcus aureus HG001 (RomR) to learn more about the resistance mechanism. We showed that Rom-resistance is based on a single point mutation in the coding region of farR [regulator of fatty acid (FA) resistance] that causes an amino acid change from Cys to Arg at position 116 in FarR, that affects FarR activity. Comparative transcriptome analysis revealed that mutated farR affects transcription of many genes in distinct pathways. FarR represses for example the expression of its own gene (farR), its flanking gene farE (effector of FA resistance), and other global regulators such as agr and sarA. All these genes were consequently upregulated in the RomR clone. Particularly the upregulation of agr and sarA leads to increased expression of virulence genes rendering the RomR clone more cytotoxic and more pathogenic in a mouse infection model. The Rom-resistance is largely due to the de-repression of farE. FarE is described as an efflux pump for linoleic and arachidonic acids. We observed an increased release of lipids in the RomR clone compared to its parental strain HG001. If farE is deleted in the RomR clone, or, if native farR is expressed in the RomR strain, the corresponding strains become hypersensitive to Rom. Overall, we show here that the high Rom-resistance is mediated by overexpression of farE in the RomR clone, that FarR is an important regulator, and that the point mutation in farR (RomR clone) makes the clone hyper-virulent.
KW  - antibiotic
KW  - Gram-positive bacteria
KW  - rhodomyrtone
KW  - Staphylococcus
KW  - membrane active
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224117
VL  - 10
ER  - 
TY  - JOUR
A1  - John, Cathy N.
A1  - Abrantes, Pedro M. D. S.
A1  - Prusty, Bhupesh K.
A1  - Ablashi, Dharam V.
A1  - Africa, Charlene W. J.
T1  - K21 Compound, a Potent Antifungal Agent: Implications for the Treatment of Fluconazole-Resistant HIV-Associated Candida Species
JF  - Frontiers in Microbiology
N2  - Background/Objectives: With mucocutaneous candidiasis being highly prevalent in HIV patients, the emergence of fluconazole-resistant Candida species forms a major challenge in treating and eradicating these infections. The objective of this study was to establish the antifungal activity of K21, a membrane-rupturing antimicrobial compound derived from a silica quaternary ammonium compound (SiQAC) with tetraethoxysilane (TEOS).

Methods: The study sample included 81 Candida species of which 9 were type strains and 72 were clinical isolates. Minimum inhibitory concentrations, synergy, fractional inhibitory concentration index (FICI), and time kill assays were determined by broth microdilution. Electron microscopy (EM) was used to determine the qualitative changes brought about after treatment with K21.

Results: K21 inhibited the growth of all fluconazole-resistant and susceptible Candida strains with only 2 h of exposure required to effectively kill 99.9% of the inoculum, and a definite synergistic effect was observed with a combination of K21 and fluconazole. EM demonstrated the presence of two forms of extracellular vesicles indicative of biofilm formation and cell lysis.

Conclusion: The study established the efficacy of K21 as an antifungal agent and with fluconazole-resistant candidiasis on the increase, the development of K21 can provide a promising alternative to combat acquired drug resistance.
KW  - HIV-associated candidiasis
KW  - antimicrobial compounds
KW  - Candida
KW  - fluconazole
KW  - antifungal susceptibility
KW  - broth microdilution
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323505
VL  - 10
ER  - 
TY  - THES
A1  - Hofmann, Kristina Simone
T1  - Untersuchung von mechanischen Eigenschaften, Bruchfestigkeit und Haftfestigkeit von Multilayer Zirkoniumdioxidkeramik zu dualhärtenden Befestigungskompositen
T1  - Investigation of mechanical properties, fracture strength and shear bond strength of multilayer zirconia ceramics to dual-curing luting composites
N2  - Ziel der Arbeit war es, den Einfluss der Platzierung der Restauration im Rohling auf die mechanischen Eigenschaften und die Scherfestigkeit zu dualhärtenden Befestigungskompositen zu untersuchen sowie Unterschiede in der Bruchfestigkeit zwischen zwei Multilayerkeramiken zu ermitteln.

Material und Methodik: Es wurden 160 zylindrische Prüfkörper aus der Multilayerkeramik Katana Zirconia ML hergestellt, um mechanische Eigenschaften wie Dichte, Biegefestigkeit und Härte zu bestimmen. Eine Gruppe wurde künstlich gealtert. Die Bruchfestigkeit von 32 Kronen (Katana Zirconia ML, Ceramill Zolid FX Multilayer) wurde vor und nach thermischer sowie mechanischer Belastung untersucht. Zur Bestimmung der Scherfestigkeit wurden 512 quadratische Prüfkörper hergestellt, die verschiedenen thermischen Belastungen ausgesetzt wurden. Die Scherfestigkeit wurde mit einer Universalprüfmaschine bestimmt und die Brucharten (adhäsiv, kohäsiv, gemischt)  wurden analysiert. 

Ergebnisse: Es gab keinen signifikanten Unterschied zwischen den Schichten oder der Alterung bei Dichte, Biegefestigkeit und Härte. Katana Zirconia ML zeigte höhere Bruchfestigkeit als Ceramill Zolid FX Multilayer. Die Scherfestigkeit von Panavia V5 zu Katana Zirconia ML war nicht durch thermische Belastung beeinflusst, jedoch bei Ceramill Zolid FX Multilayer. Der Haftverbund von Visalys CemCore war durch thermische Belastung beeinflusst, während Panavia V5 zu beiden Keramiken höhere Werte aufwies. Katana Zirconia ML hatte höhere Scherfestigkeitswerte als Ceramill Zolid FX Multilayer.

Schlussfolgerung: Multilayerkeramik stellt eine arbeitsverringernde Alternative für den Seitenzahnbereich dar, ohne mechanische und ästhetische Einbußen. Die Platzierung im Rohling hat keinen Einfluss auf die Eigenschaften, jedoch ist der Haftverbund vom Befestigungskomposit abhängig.
N2  - The aim of the work was to investigate the influence of the placement of the restoration in the blank on the mechanical properties and the shear bond strength of dual-curing luting composites and to determine differences in the fracture strength between two multilayer ceramics.

Material and method: 160 cylindrical test specimens were made from the multilayer ceramic Katana Zirconia ML to determine mechanical properties such as density, flexural strength and hardness. One group was artificially aged. The breaking strength of 32 crowns (Katana Zirconia ML, Ceramill Zolid FX Multilayer) was examined before and after thermal and mechanical loading. To determine the shear bond strength, 512 square test specimens were produced and subjected to various thermal loads. The shear bond strength was determined using a universal testing machine and the fracture types (adhesive, cohesive, mixed) were analyzed. 

Results: There was no significant difference between layers or aging in density, flexural strength and hardness. Katana Zirconia ML showed higher breaking strength than Ceramill Zolid FX Multilayer. The shear bond strength of Panavia V5 to Katana Zirconia ML was not affected by thermal loading, but was affected in combination with Ceramill Zolid FX Multilayer. The adhesive bond of Visalys CemCore was influenced by thermal stress, while Panavia V5 had higher shear bond strength to both ceramics. Katana Zirconia ML had higher shear bond strength than Ceramill Zolid FX Multilayer.

Conclusion: Multilayer ceramics represent a labor-reducing alternative for the posterior region, without mechanical and aesthetic losses. The placement in the blank has no influence on the properties, but the adhesive bond depends on the fixing composite.
KW  - Zirkoniumoxidkeramik
KW  - Multilayer-Zirkoniumdioxidkeramiken
KW  - Biaxiale Biegefestigkeit
KW  - Vickers-Härte
KW  - CAD/CAM restoration material
KW  - adhesive luting agent
KW  - shear bond strength
KW  - Biegefestigkeit
KW  - Härte
KW  - Mechanische Eigenschaft
KW  - Restaurative Zahnmedizin
KW  - Scherfestigkeit
KW  - Befestigungszement
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370308
ER  - 
TY  - JOUR
A1  - Gronwald, Thomas
A1  - Hoos, Olaf
A1  - Hottenrott, Kuno
T1  - Effects of Acute Normobaric Hypoxia on Non-linear Dynamics of Cardiac Autonomic Activity During Constant Workload Cycling Exercise
JF  - Frontiers in Physiology
N2  - Aim: Measurements of Non-linear dynamics of heart rate variability (HRV) provide new possibilities to monitor cardiac autonomic activity during exercise under different environmental conditions. Using detrended fluctuation analysis (DFA) technique to assess correlation properties of heart rate (HR) dynamics, the present study examines the influence of normobaric hypoxic conditions (HC) in comparison to normoxic conditions (NC) during a constant workload exercise.

Materials and Methods: Nine well trained cyclists performed a continuous workload exercise on a cycle ergometer with an intensity corresponding to the individual anaerobic threshold until voluntary exhaustion under both NC and HC (15% O2). The individual exercise duration was normalized to 10% sections (10–100%). During exercise HR and RR-intervals were continuously-recorded. Besides HRV time-domain measurements (meanRR, SDNN), fractal correlation properties using short-term scaling exponent alpha1 of DFA were calculated. Additionally, blood lactate (La), oxygen saturation of the blood (SpO2), and rating of perceived exertion (RPE) were recorded in regular time intervals.

Results: We observed significant changes under NC and HC for all parameters from the beginning to the end of the exercise (10% vs. 100%) except for SpO2 and SDNN during NC: increases for HR, La, and RPE in both conditions; decreases for SpO2 and SDNN during HC, meanRR and DFA-alpha1 during both conditions. Under HC HR (40–70%), La (10–90%), and RPE (50–90%) were significantly-higher, SpO2 (10–100%), meanRR (40–70%), and DFA-alpha1 (20–60%) were significantly-lower than under NC.

Conclusion: Under both conditions, prolonged exercise until voluntary exhaustion provokes a lower total variability combined with a reduction in the amplitude and correlation properties of RR fluctuations which may be attributed to increased organismic demands. Additionally, HC provoked higher demands and loss of correlation properties at an earlier stage during the exercise regime, implying an accelerated alteration of cardiac autonomic regulation.
KW  - autonomic nervous system
KW  - heart rate variability
KW  - detrended fluctuation analysis
KW  - endurance exercise
KW  - voluntary exhaustion
KW  - hypoxia
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369199
VL  - 10
ER  - 
TY  - JOUR
A1  - Fahmy-Garcia, Shorouk
A1  - Farrell, Eric
A1  - Witte-Bouma, Janneke
A1  - Robbesom-van den Berge, Iris
A1  - Suarez, Melva
A1  - Mumcuoglu, Didem
A1  - Walles, Heike
A1  - Kluijtmans, Sebastiaan G. J. M.
A1  - van der Eerden, Bram C. J.
A1  - van Osch, Gerjo J. V. M.
A1  - van Leeuwen, Johannes P. T. M.
A1  - van Driel, Marjolein
T1  - Follistatin Effects in Migration, Vascularization, and Osteogenesis in vitro and Bone Repair in vivo
JF  - Frontiers in Bioengineering and Biotechnology
N2  - The use of biomaterials and signaling molecules to induce bone formation is a promising approach in the field of bone tissue engineering. Follistatin (FST) is a glycoprotein able to bind irreversibly to activin A, a protein that has been reported to inhibit bone formation. We investigated the effect of FST in critical processes for bone repair, such as cell recruitment, osteogenesis and vascularization, and ultimately its use for bone tissue engineering. In vitro, FST promoted mesenchymal stem cell (MSC) and endothelial cell (EC) migration as well as essential steps in the formation and expansion of the vasculature such as EC tube-formation and sprouting. FST did not enhance osteogenic differentiation of MSCs, but increased committed osteoblast mineralization. In vivo, FST was loaded in an in situ gelling formulation made by alginate and recombinant collagen-based peptide microspheres and implanted in a rat calvarial defect model. Two FST variants (FST288 and FST315) with major differences in their affinity to cell-surface proteoglycans, which may influence their effect upon in vivo bone repair, were tested. In vitro, most of the loaded FST315 was released over 4 weeks, contrary to FST288, which was mostly retained in the biomaterial. However, none of the FST variants improved in vivo bone healing compared to control. These results demonstrate that FST enhances crucial processes needed for bone repair. Further studies need to investigate the optimal FST carrier for bone regeneration.
KW  - follistatin 315 (FST315)
KW  - follistatin 288 (FST288)
KW  - migration
KW  - vascularization
KW  - osteogenesis
KW  - injectable in situ gelling slow release system
KW  - bone tissue engineering
KW  - regenerative medicine
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227617
VL  - 7
ER  - 
TY  - JOUR
A1  - Gál, Bernadett I.
A1  - Kilencz, Tünde
A1  - Albert, Anita
A1  - Demeter, Ildikó
A1  - Hegedűs, Klára Mária
A1  - Janka, Zoltán
A1  - Csifcsák, Gábor
A1  - Álmos, Péter Z.
T1  - Mild Effect of Nalmefene on Alcoholic Cue-Induced Response Invigoration in Alcohol Use Disorder Without Accompanying Changes in Electrophysiological Signatures of Early Visual Processing and Executive Control
JF  - Frontiers in Pharmacology
N2  - Nalmefene is approved for as-needed pharmacological treatment in alcohol use disorder (AUD) by the European Medicines Agency. While the cellular effects of nalmefene have been thoroughly investigated, data are very limited on how this agent influences neural signals associated with inhibitory control and the visual analysis of environmental cues. This double-blind crossover study assessed the behavioral and neural effects of acute nalmefene administration in patients diagnosed with AUD. In experiment 1, we validated our experimental paradigm (electroencephalography combined with a modified Go/NoGo task using images of alcoholic and nonalcoholic drinks as prime stimuli) in 20 healthy adults to ensure that our protocol is suitable for assessing the behavioral and neural aspects of executive control. In experiment 2, we recruited 19 patients with AUD, and in a double-blind crossover design, we investigated the effects of nalmefene versus placebo on task performance (response accuracy, the sensitivity index, and reaction times), visual responses to appetitive cues (occipital P1, N1, and P2 components), and electrophysiological markers of conflict detection and response inhibition (frontal N2 and P3 waveforms). Under placebo, patients produced faster reaction times to alcohol-primed Go stimuli, an effect that was weak despite being statistically significant. However, the effect of alcoholic cues on the speed of response initiation disappeared after receiving nalmefene. We found no placebo versus nalmefene difference regarding our patients’ ability to accurately inhibit responses to NoGo stimuli or for occipital and frontal event-related potentials. Our results suggest that nalmefene might be potent in reducing the vigor to act upon alcoholic cues in AUD patients, but this effect is most probably mediated via subcortical (rather than cortical) neural circuits.
KW  - nalmefene
KW  - alcohol use disorder
KW  - response inhibition
KW  - incentive salience
KW  - event-related potentials
KW  - Go/NoGo task
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369182
VL  - 10
ER  - 
TY  - JOUR
A1  - Franke, Maximilian
A1  - Conzelmann, Annette
A1  - Grünblatt, Edna
A1  - Werling, Anna M.
A1  - Spieles, Helen
A1  - Wewetzer, Christoph
A1  - Warnke, Andreas
A1  - Romanos, Marcel
A1  - Walitza, Susanne
A1  - Renner, Tobias J.
T1  - No Association of Variants of the NPY-System With Obsessive-Compulsive Disorder in Children and Adolescents
JF  - Frontiers in Molecular Neuroscience
N2  - Obsessive-compulsive disorder (OCD) causes severe distress and is therefore counted by the World Health Organisation (WHO) as one of the 10 most impairing illnesses. There is evidence for a strong genetic underpinning especially in early onset OCD (eoOCD). Though several genes involved in neurotransmission have been reported as candidates, there is still a need to identify new pathways. In this study, we focussed on genetic variants of the Neuropeptide Y (NPY) system. NPY is one of the most abundant neuropeptides in the human brain with emerging evidence of capacity to modulate stress response, which is of high relevance in OCD. We focussed on tag-SNPs of NPY and its receptor gene NPY1R in a family-based approach. The sample comprised 86 patients (children and adolescents) with eoOCD with both their biological parents. However, this first study on genetic variants of the NPY-system could not confirm the association between the investigated SNPs and eoOCD. Based on the small sample size results have to be interpreted as preliminary and should be replicated in larger samples. However, also in an additional GWAS analysis in a large sample, we could not observe an associations between NPY and OCD. Overall, these preliminary results point to a minor role of NPY on the stress response of OCD.
KW  - NPY
KW  - obsessive-compulsive
KW  - children
KW  - anxiety
KW  - neuropeptide
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229051
VL  - 12
ER  - 
TY  - JOUR
A1  - Geran, Rohat
A1  - Uecker, Florian C.
A1  - Prüss, Harald
A1  - Haeusler, Karl Georg
A1  - Paul, Friedemann
A1  - Ruprecht, Klemens
A1  - Harms, Lutz
A1  - Schmidt, Felix A.
T1  - Olfactory and Gustatory Dysfunction in Patients With Autoimmune Encephalitis
JF  - Frontiers in Neurology
N2  - Objective: To test the hypothesis that olfactory (OF) and gustatory function (GF) is disturbed in patients with autoimmune encephalitides (AE).

Methods: The orthonasal OF was tested in 32 patients with AE and 32 age- and sex-matched healthy controls (HC) with the standardized Threshold Discrimination Identification (TDI) score. This validated olfactory testing method yields individual scores for olfactory threshold (T), odor discrimination (D), and identification (I), along with a composite TDI score. The GF was determined by the Taste Strip Test (TST).

Results: Overall, 24/32 (75%) of patients with AE, but none of 32 HC (p < 0.001) had olfactory dysfunction in TDI testing. The results of the threshold, discrimination and identification subtests were significantly reduced in patients with AE compared to HC (all p < 0.001). Assessed by TST, 5/19 (26.3%) of patients with AE, but none of 19 HC presented a significant limitation in GF (p < 0.001). The TDI score was correlated with the subjective estimation of the olfactory capacity on a visual analog scale (VAS; rs = 0.475, p = 0.008). Neither age, sex, modified Rankin Scale nor disease duration were associated with the composite TDI score.

Conclusions: This is the first study investigating OF and GF in AE patients. According to unblinded assessment, patients with AE have a reduced olfactory and gustatory capacity compared to HC, suggesting that olfactory and gustatory dysfunction are hitherto unrecognized symptoms in AE. Further studies with larger number of AE patients would be of interest to verify our results.
KW  - autoimmune encephalitis
KW  - olfactory dysfunction
KW  - gustatory dysfunction
KW  - olfactory testing
KW  - threshold discrimination identification test
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232921
VL  - 10
ER  - 
TY  - JOUR
A1  - Jarick, Katja J.
A1  - Mokhtari, Zeinab
A1  - Scheller, Lukas
A1  - Hartweg, Julia
A1  - Thusek, Sina
A1  - Le, Duc-Dung
A1  - Ranecky, Maria
A1  - Shaikh, Haroon
A1  - Qureischi, Musga
A1  - Heinze, Katrin G.
A1  - Beilhack, Andreas
T1  - Photoconversion of Alloreactive T Cells in Murine Peyer’s Patches During Acute Graft-Versus-Host Disease: Tracking the Homing Route of Highly Proliferative Cells In Vivo
JF  - Frontiers in Immunology
N2  - The regulation of immune cell migration throughout the body is essential to warrant immunosurveillance and to maintain immune homeostasis. Marking and tracking of these cells has proven important to study mechanisms of immune cell trafficking and cell interaction in vivo. Photoconversion is a well-suited technique for intravital application because it enables contactless time- and location-specific marking of cells in the tissue without surgically manipulating the microenvironment of the cells in question. However, in dividing cells the converted fluorescent protein may decline quickly. Here, we provide a detailed description of the photoconversion technique and its applicability to tracking highly proliferating T cells from the priming site of T cell activation to peripheral target organs of effector function in a preclinical model. Dendra2+ T cells were photoconverted in the Peyer’s patches during the initiation phase of acute graft-versus-host disease (GvHD) and tracked through the mesenteric lymph nodes and the peripheral blood to the small intestine with flow cytometry and intravital two-photon microscopy. Photoconverted alloreactive T cells preserved the full proliferative capacity, homing, and migration of alloreactive T cells in the intestinal lamina propria. We conclusively proved that photoconversion of highly proliferative alloreactive T cells in the Peyer’s patches is an effective tool to study trafficking of alloreactive T cells under physiologic conditions and to GvHD target tissues. This technique can also be applied to the study of immune cell tracking under inflammatory and non-inflammatory conditions.
KW  - T cell migration
KW  - acute graft-versus-host disease
KW  - mouse models
KW  - photoconversion
KW  - Dendra2
KW  - Peyer's patch
KW  - in vivo cell tracking
KW  - lymphocyte homing
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323309
VL  - 9
ER  - 
TY  - JOUR
A1  - Herster, Franziska
A1  - Bittner, Zsofia
A1  - Codrea, Marius Cosmin
A1  - Archer, Nathan K.
A1  - Heister, Martin
A1  - Löffler, Markus W.
A1  - Heumos, Simon
A1  - Wegner, Joanna
A1  - Businger, Ramona
A1  - Schindler, Michael
A1  - Stegner, David
A1  - Schäkel, Knut
A1  - Grabbe, Stephan
A1  - Ghoreschi, Kamran
A1  - Miller, Lloyd S.
A1  - Weber, Alexander N. R.
T1  - Platelets Aggregate With Neutrophils and Promote Skin Pathology in Psoriasis
JF  - Frontiers in Immunology
N2  - Psoriasis is a frequent systemic inflammatory autoimmune disease characterized primarily by skin lesions with massive infiltration of leukocytes, but frequently also presents with cardiovascular comorbidities. Especially polymorphonuclear neutrophils (PMNs) abundantly infiltrate psoriatic skin but the cues that prompt PMNs to home to the skin are not well-defined. To identify PMN surface receptors that may explain PMN skin homing in psoriasis patients, we screened 332 surface antigens on primary human blood PMNs from healthy donors and psoriasis patients. We identified platelet surface antigens as a defining feature of psoriasis PMNs, due to a significantly increased aggregation of neutrophils and platelets in the blood of psoriasis patients. Similarly, in the imiquimod-induced experimental in vivo mouse model of psoriasis, disease induction promoted PMN-platelet aggregate formation. In psoriasis patients, disease incidence directly correlated with blood platelet counts and platelets were detected in direct contact with PMNs in psoriatic but not healthy skin. Importantly, depletion of circulating platelets in mice in vivo ameliorated disease severity significantly, indicating that both PMNs and platelets may be relevant for psoriasis pathology and disease severity.
KW  - psoriasis
KW  - neutrophil
KW  - platelet
KW  - platelet-neutrophil complexes (PNCs)
KW  - imiquimod
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-320175
VL  - 10
ER  - 
TY  - JOUR
A1  - Kasaragod, Vikram B.
A1  - Schindelin, Hermann
T1  - Structure–Function Relationships of Glycine and GABAA Receptors and Their Interplay With the Scaffolding Protein Gephyrin
JF  - Frontiers in Molecular Neuroscience
N2  - Glycine and γ-aminobutyric acid (GABA) are the major determinants of inhibition in the central nervous system (CNS). These neurotransmitters target glycine and GABAA receptors, respectively, which both belong to the Cys-loop superfamily of pentameric ligand-gated ion channels (pLGICs). Interactions of the neurotransmitters with the cognate receptors result in receptor opening and a subsequent influx of chloride ions, which, in turn, leads to hyperpolarization of the membrane potential, thus counteracting excitatory stimuli. The majority of glycine receptors and a significant fraction of GABAA receptors (GABAARs) are recruited and anchored to the post-synaptic membrane by the central scaffolding protein gephyrin. This ∼93 kDa moonlighting protein is structurally organized into an N-terminal G-domain (GephG) connected to a C-terminal E-domain (GephE) via a long unstructured linker. Both inhibitory neurotransmitter receptors interact via a short peptide motif located in the large cytoplasmic loop located in between transmembrane helices 3 and 4 (TM3-TM4) of the receptors with a universal receptor-binding epitope residing in GephE. Gephyrin engages in nearly identical interactions with the receptors at the N-terminal end of the peptide motif, and receptor-specific interaction toward the C-terminal region of the peptide. In addition to its receptor-anchoring function, gephyrin also interacts with a rather large collection of macromolecules including different cytoskeletal elements, thus acting as central scaffold at inhibitory post-synaptic specializations. Dysfunctions in receptor-mediated or gephyrin-mediated neurotransmission have been identified in various severe neurodevelopmental disorders. Although biochemical, cellular and electrophysiological studies have helped to understand the physiological and pharmacological roles of the receptors, recent high resolution structures of the receptors have strengthened our understanding of the receptors and their gating mechanisms. Besides that, multiple crystal structures of GephE in complex with receptor-derived peptides have shed light into receptor clustering by gephyrin at inhibitory post-synapses. This review will highlight recent biochemical and structural insights into gephyrin and the GlyRs as well as GABAA receptors, which provide a deeper understanding of the molecular machinery mediating inhibitory neurotransmission.
KW  - glycine receptors
KW  - GABAA receptors
KW  - gephyrin
KW  - moonlighting protein
KW  - inhibitory post-synaptic specialization
KW  - cytoskeletal proteins
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325607
VL  - 11
ER  - 
TY  - JOUR
A1  - Kervarrec, Thibault
A1  - Samimi, Mahtab
A1  - Guyétant, Serge
A1  - Sarma, Bhavishya
A1  - Chéret, Jérémy
A1  - Blanchard, Emmanuelle
A1  - Berthon, Patricia
A1  - Schrama, David
A1  - Houben, Roland
A1  - Touzé, Antoine
T1  - Histogenesis of Merkel Cell Carcinoma: A Comprehensive Review
JF  - Frontiers in Oncology
N2  - Merkel cell carcinoma (MCC) is a primary neuroendocrine carcinoma of the skin. This neoplasia features aggressive behavior, resulting in a 5-year overall survival rate of 40%. In 2008, Feng et al. identified Merkel cell polyomavirus (MCPyV) integration into the host genome as the main event leading to MCC oncogenesis. However, despite identification of this crucial viral oncogenic trigger, the nature of the cell in which MCC oncogenesis occurs is actually unknown. In fact, several hypotheses have been proposed. Despite the large similarity in phenotype features between MCC tumor cells and physiological Merkel cells (MCs), a specialized subpopulation of the epidermis acting as mechanoreceptor of the skin, several points argue against the hypothesis that MCC derives directly from MCs. Alternatively, MCPyV integration could occur in another cell type and induce acquisition of an MC-like phenotype. Accordingly, an epithelial as well as a fibroblastic or B-cell origin of MCC has been proposed mainly based on phenotype similarities shared by MCC and these potential ancestries. The aim of this present review is to provide a comprehensive review of the current knowledge of the histogenesis of MCC.
KW  - merkel cell polyomavirus (MCPyV)
KW  - epithelial
KW  - fibroblast
KW  - B cell
KW  - Merkel cell carcinoma (MCC)
KW  - histogenesis
KW  - origin
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325733
VL  - 9
ER  - 
TY  - JOUR
A1  - Nagy, Magdolna
A1  - van Geffen, Johanna P.
A1  - Stegner, David
A1  - Adams, David J.
A1  - Braun, Attila
A1  - de Witt, Susanne M.
A1  - Elvers, Margitta
A1  - Geer, Mitchell J.
A1  - Kuijpers, Marijke J. E.
A1  - Kunzelmann, Karl
A1  - Mori, Jun
A1  - Oury, Cécile
A1  - Pircher, Joachim
A1  - Pleines, Irina
A1  - Poole, Alastair W.
A1  - Senis, Yotis A.
A1  - Verdoold, Remco
A1  - Weber, Christian
A1  - Nieswandt, Bernhard
A1  - Heemskerk, Johan W. M.
A1  - Baaten, Constance C. F. M. J.
T1  - Comparative Analysis of Microfluidics Thrombus Formation in Multiple Genetically Modified Mice: Link to Thrombosis and Hemostasis
JF  - Frontiers in Cardiovascular Medicine
N2  - Genetically modified mice are indispensable for establishing the roles of platelets in arterial thrombosis and hemostasis. Microfluidics assays using anticoagulated whole blood are commonly used as integrative proxy tests for platelet function in mice. In the present study, we quantified the changes in collagen-dependent thrombus formation for 38 different strains of (genetically) modified mice, all measured with the same microfluidics chamber. The mice included were deficient in platelet receptors, protein kinases or phosphatases, small GTPases or other signaling or scaffold proteins. By standardized re-analysis of high-resolution microscopic images, detailed information was obtained on altered platelet adhesion, aggregation and/or activation. For a subset of 11 mouse strains, these platelet functions were further evaluated in rhodocytin- and laminin-dependent thrombus formation, thus allowing a comparison of glycoprotein VI (GPVI), C-type lectin-like receptor 2 (CLEC2) and integrin α6β1 pathways. High homogeneity was found between wild-type mice datasets concerning adhesion and aggregation parameters. Quantitative comparison for the 38 modified mouse strains resulted in a matrix visualizing the impact of the respective (genetic) deficiency on thrombus formation with detailed insight into the type and extent of altered thrombus signatures. Network analysis revealed strong clusters of genes involved in GPVI signaling and Ca2+ homeostasis. The majority of mice demonstrating an antithrombotic phenotype in vivo displayed with a larger or smaller reduction in multi-parameter analysis of collagen-dependent thrombus formation in vitro. Remarkably, in only approximately half of the mouse strains that displayed reduced arterial thrombosis in vivo, this was accompanied by impaired hemostasis. This was also reflected by comparing in vitro thrombus formation (by microfluidics) with alterations in in vivo bleeding time. In conclusion, the presently developed multi-parameter analysis of thrombus formation using microfluidics can be used to: (i) determine the severity of platelet abnormalities; (ii) distinguish between altered platelet adhesion, aggregation and activation; and (iii) elucidate both collagen and non-collagen dependent alterations of thrombus formation. This approach may thereby aid in the better understanding and better assessment of genetic variation that affect in vivo arterial thrombosis and hemostasis.
KW  - arterial thrombus formation
KW  - bleeding
KW  - collagen
KW  - glycoprotein VI
KW  - platelets
KW  - microfluidics
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232194
VL  - 6
ER  - 
TY  - THES
A1  - Zhang, Tengyu
T1  - Development of Modified polylysine based antibody conjugated nanoparticles with tumor-restricted, FcγR-independent stimulatory activity by targeting Fn14
T1  - Entwicklung modifizierter, mit Antikörpern konjugierter
Nanopartikel auf Polylysinbasis mit tumorbeschränkter, FcγR-
unabhängiger stimulierender Aktivität durch Ausrichtung auf Fn14
N2  - In this study, we developed an innovative nanoparticle formulation to facilitate the delivery of antitumor antibodies to tumor sites. The study commenced with the utilization of 13 bispecific antibody fusion proteins, which targeted the Fn14 receptor, thereby validating the pivotal role of crosslinking in Fn14 receptor activation. Subsequently, gold nanoparticles were activated using COOH-PEG-SH in combination with EDC/NHS, and subsequently conjugated with two Fn14-targeting antibodies, PDL192 and 5B6. Following this, a pH-sensitive shell was generated on the outer layer of the antibody-coupled gold nanoparticles through the application of chemically modified polylysine. The resultant complexes, termed MPL-antibody-AuNP, demonstrated a release profile reminiscent of the tumor microenvironment (TME). Notably, these complexes released antibody-AuNPs only in slightly acidic conditions while remaining intact in neutral or basic environments. Functionality analysis further affirmed the pH-sensitive property of MPL-antibody-AuNPs, demonstrating that the antibodies only initiated potent Fn14 activation in slightly acidic environments. This formulation holds potential for applicability to antibodies or ligands targeting the 80 TNFRSF family, given that gold nanoparticles successfully served as platforms for antibody crosslinking, thereby transforming these antibodies into potent agonists. Moreover, the TME disintegration profile of MPL mitigates the potential cytotoxic effects of antibodies, thereby circumventing associated adverse side effects. This study not only showcases the potential of nanoparticle formulations in targeted therapy, but also provides a solid foundation for further investigations on their clinical application in the context of targeting category II TNFRSF receptors with antibodies or ligands.
N2  - In dieser Studie haben wir eine innovative Nanopartikel-Formulierung entwickelt, um die Auslieferung von Antitumor-Antikörpern an Tumorstellen zu erleichtern. Die Studie begann mit der Verwendung von 13 bispezifischen Antikörper-Fusionsproteinen, die auf den Fn14-Rezeptor abzielten, wodurch die entscheidende Rolle der Quervernetzung bei der Aktivierung des Fn14-Rezeptors bestätigt wurde. Anschließend wurden Goldnanopartikel unter Verwendung von COOH-PEG-SH in Kombination mit EDC/NHS aktiviert und danach mit zwei auf Fn14 abzielenden Antikörpern, PDL192 und 5B6, konjugiert. Daraufhin wurde eine pH-sensitive Schale auf der äußeren Schicht der mit Antikörpern gekoppelten Goldnanopartikel durch den Einsatz von chemisch modifiziertem Polylysin erzeugt. Die resultierenden Komplexe, bezeichnet als MPL-Antikörper-AuNP, zeigten ein Freisetzungsprofil, das an das Tumormikroumfeld (TME) erinnert. Bemerkenswert ist, dass diese Komplexe Antikörper-AuNP nur in leicht sauren Bedingungen freisetzten, während sie in neutralen oder basischen Umgebungen intakt blieben. Die Funktionalitätsanalyse bestätigte weiterhin die pH-empfindliche Eigenschaft der MPL-Antikörper-AuNPs, was zeigt, dass die Antikörper eine potente Fn14-Aktivierung nur in leicht sauren Bedingungen initiierten. Diese Formulierung hat Potenzial für die Anwendbarkeit auf Antikörper oder Liganden, die auf die Familie der TNFRSF abzielen, da die Goldnanopartikel erfolgreich als Plattformen für die Antikörpervernetzung dienten und diese Antikörper in potente Agonisten verwandelten. Darüber hinaus mildert das TME-Zerfallsprofil von MPL die potenziellen zytotoxischen Effekte der Antikörper, wodurch die damit verbundenen negativen Nebenwirkungen umgangen werden. Diese Studie zeigt nicht nur das Potenzial von Nanopartikel-Formulierungen in der gezielten Therapie auf, sondern bietet auch eine solide Grundlage für weitere Untersuchungen zu ihrer klinischen Anwendung im Kontext der Zielrichtung auf Kategorie-II-TNFRSF-Rezeptoren mit Antikörpern oder Liganden.
KW  - Immuntherapie
KW  - Immunotherapy
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358650
ER  - 
TY  - THES
A1  - Hadi, Naji Said Aboud
T1  - In vitro Studies on the Genotoxicity of Selected Pyrrolizidine Alkaloids
T1  - In-vitro-Studien zur Genotoxizität ausgewählter Pyrrolizidinalkaloide
N2  - Cancer is one of the leading causes of death worldwide. Toxic contaminants in human food or medicinal products, such as substances like pyrrolizidine alkaloids (PAs), have been thought to contribute to cancer incidence. PAs are found in many plant species as secondary metabolites, and they may affect humans through contaminated food sources, herbal medicines, and dietary supplements. Hundreds of compounds belonging to PAs have been identified, differing in their chemical structures, either in their necine base moiety or esterification at their necic acid moiety. PAs undergo hepatic metabolism, and after this process, they can induce hepatotoxicity, genotoxicity, and carcinogenicity. However, the mechanism of inducing genotoxicity and carcinogenicity is still unclear and warrants further investigation.
Therefore, the present study aims to investigate the mechanism of genotoxicity induced by selected PAs with different chemical structures in in vitro systems. Primarily, human hepatoma HepG2 cells were utilized, and in co-culture, metabolically active HepG2 cells were combined with non-metabolically active human cervical HeLa H2B-GFP cells.

First, the genotoxicity of the PAs europine, lycopsamine, retrorsine, riddelliine, seneciphylline, echimidine, and lasiocarpine was investigated in the cytokinesis-block micronucleus (CBMN) assay. All seven selected PAs caused the formation of micronuclei in a dose-dependent manner, with the maximal increase of micronucleus formation ranging from 1.64 to 2.0 fold. The lowest concentrations at which significant induction of micronuclei was found were 3.2 µM for lasiocarpine and riddelliine, 32 µM for retrorsine and echimidine, and 100 µM for seneciphylline, europine, and lycopsamine. These results confirmed previously published potency rankings in the micronucleus assay.
The same PAs, with the exception of seneciphylline, were also investigated in a crosslink-modified comet assay, and reduced tail formation after hydrogen peroxide treatment was found in all diester-type PAs. Meanwhile, an equimolar concentration of the monoesters europine and lycopsamine did not significantly reduce DNA migration. Thus, the crosslinking activity was related to the ester type.

Next, the role of metabolic enzymes and membrane transporters in PA-induced genotoxicity was assessed. Ketoconazole (CYP 450-3A4 inhibitor) prevented lasiocarpine-induced micronucleus formation completely, while furafylline (CYP 450-1A2 inhibitor) reduced lasiocarpine-induced micronucleus formation, but did not abolish it completely. This implies that the CYP 450 enzymes play an important role in PA-induced genotoxicity. 
Carboxylesterase 2 enzyme (CES 2) is commonly known to be involved in the detoxification of xenobiotics. Loperamide (CES 2 inhibitor) yielded an increased formation of lasiocarpine-induced micronuclei, revealing a possible role of CES-mediated detoxification in the genotoxicity of lasiocarpine. Also, intracellular glutathione (GSH) plays an important role in the detoxification of xenobiotics or toxins in the cells. Cells which had been pretreated with L-buthionine sulfoximine (BSO) to reduce GSH content were significantly more sensitive for the induction of micronucleus formation by lasiocarpine revealing the importance of GSH in PA-induced genotoxicity.
 
Quinidine (Q) and nelfinavir (NFR) are OCT1 and OATP1B1 influx transporter inhibitors, respectively, which reduced micronucleus induction by lasiocarpine (only quinidine significantly), but not completely, pointing to a relevance of OCT1 for PA uptake in HepG2 cells. Verapamil (V) and benzbromarone (Bz) are MDR1 and MRP2 efflux transporter inhibitors, respectively, and they caused a slightly increased micronucleus induction by lasiocarpine (significant only for benzbromarone) thus, revealing the role of efflux transporters in PA-induced genotoxicity. 

The mechanistic approach to PA-induced genotoxicity was further studied based on oxidative stress via the formation of reactive oxygen species (ROS) in HepG2 cells. Overproduction of ROS can cross-link cellular macromolecules such as DNA, leading to genomic damage. An equimolar concentration of 10 µM of lasiocarpine (open-diester PA), riddelliine (cyclic-diester PA), and europine (monoester) significantly induced ROS production, with the highest ROS generation observed after lasiocarpine treatment, followed by riddelliine and then europine. No significant increase in ROS production was found with lycopsamine (10 µM; monoester PA), even at a higher concentration (320 µM). The generation of ROS by these PAs was further analyzed for confirmation by using 5 mM of the thiol radical scavenger antioxidant N-acetyl cysteine (NAC) combined with lasiocarpine, riddelliine, or europine. This analysis yielded a significant decrease in ROS after combining NAC with lasiocarpine, riddelliine, and europine. In addition, lasiocarpine, riddelliine, and europine induced a loss of mitochondrial membrane potential, pointing to mitochondria as the source of ROS generation.

In vivo, hepatic sinusoidal epithelial cells (HSECs) are known to be damaged first by PAs after hepatic metabolization, but HSECs themselves do not express the required metabolic enzymes for activation of PAs. To mimic this situation, HepG2 cells were used to metabolically activate PA in a co-culture with HeLa H2B-GFP cells as non-metabolically active neighbours. Due to the green fluorescent GFP label the HeLa cells could be identified easily based in the co-culture. The PAs europine, riddelliine and lasiocarpine induced micronucleus formation in HepG2 cells, and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Metabolic inhibition of CYP 450 enzymes with ketoconazole abrogated micronucleus formation induced by the same PAs tested in the co-culture. The efflux transporter inhibitors verapamil and benzbromarone reduced the micronucleus formation in the co-culture. Furthermore, mitotic disturbances as an additional genotoxic mechanism of action were observed in HepG2 cells and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Overall, we were able to show that PAs were activated by HepG2 cells and the metabolites induced genomic damage in co-cultured non-metabolically active green HeLa cells.

Finally, in HepG2 cells as well as the co-culture, combinations of PAs lasiocarpine and riddelliine favoured an additive effect rather than synergism. Thus, this study therefore provides support that the assumption of dose-addition can be applied in the characterization of the genotoxicity risk of PAs present in a mixture.
N2  - Krebs ist eine der häufigsten Todesursachen weltweit. Toxische Verunreinigungen in Lebensmitteln oder pflanzlichen Arzneimitteln, wie Pyrrolizidinalkaloide (PAs), können zur Krebsinzidenz beitragen. PAs kommen in vielen Pflanzenarten als Sekundärmetabolite vor. Menschen können diese über kontaminierte Nahrungsquellen, pflanzliche Arzneimittel und Nahrungsergänzungsmittel aufnehmen. Eine Vielzahl von Verbindungen, die zu pyrrolizidinalkaloidhaltigen Substanzen (PAs) gehören, wurden identifiziert. Diese unterscheiden sich in ihrer chemischen Struktur entweder durch ihre Necinbaseneinheit oder ihre Veresterung an der Necicsäureeinheit. Nach metabolischer Aktivierung in der Leber können PAs Hepatotoxizität, Genotoxizität und Karzinogenität induzieren. Jedoch ist der Genotoxizitätsmechanismus nicht vollständig aufgeklärt und erfordert weitere Untersuchungen.
Das Ziel dieser Studie liegt in der Untersuchung des Mechanismus der Genotoxizität, die in vitro durch bestimmte PAs mit unterschiedlicher chemischer Struktur induziert wird. Hierbei wurden primär humane Hepatom-HepG2-Zellen verwendet sowie in Co-Kultur metabolisch aktive HepG2-Zellen und nicht-metabolisch aktive humane zervikale HeLa H2B-GFP-Zellen.
Zunächst wurde die Genotoxizität der PAs Europin, Lycopsamin, Retrorsin, Riddelliin, Seneciphyllin, Echimidin und Lasiocarpin im Zytokinese-Block-Mikronukleus-Assay (CBMN) untersucht. Die sieben (7) ausgewählten PAs führten dosisabhängig zur Bildung von Mikrokernen. Der maximale Anstieg der Mikronukleusbildung lag für alle PAs im Bereich des 1,64- bis 2,0-fachen des Ausgangswertes. Die niedrigsten Konzentrationen, bei denen eine signifikante Induktion von Mikrokernen gefunden wurde, waren 3,2 μM für Lasiocarpin und Riddelliin, 32 μM für Retrorsin und Echimidin sowie 100 μM für Seneciphyllin, Europin und Lycopsamin. Diese Ergebnisse bestätigen zuvor veröffentlichte Potenz-Rankings im Mikronukleus-Assay.

Die Genotoxizität der gleichen PAs, mit Ausnahme von Seneciphyllin, wurde zusätzlich mittels eines Crosslink-modifizierten Comet-Assay untersucht. Es wurde eine reduzierte Schweifbildung nach der Behandlung mit Wasserstoffperoxid in allen PAs des Diestertyps gefunden, während eine äquimolare Konzentration der Monoester Europin und Lycopsamin die DNA-Migration nicht signifikant reduzierte. Dies deutet darauf hin, dass die Vernetzungsaktivität von PAs auf der Ester-Einheit beruht.
Als nächstes wurde die Rolle von Stoffwechselenzymen und Membrantransportern in der PA-induzierten Genotoxizität untersucht. Ketoconazol (CYP 450-3A4-Inhibitor) verhinderte die Lasiocarpin-induzierte Mikronukleusbildung vollständig, während Furafyllin (CYP 450-1A2-Inhibitor) die Lasiocarpin-induzierte Mikronukleusbildung reduzierte, aber nicht vollständig beseitigte. Dies deutet darauf hin, dass CYP 450-Enzyme eine wichtige Rolle bei der PA-induzierten Genotoxizität spielen.

Es ist allgemein bekannt, dass das Enzym Carboxylesterase 2 (CES-2) an der Entgiftung von Xenobiotika beteiligt ist. Loperamid (CES-2-Inhibitor) führte zu einer erhöhten Bildung von Lasiocarpin-induzierten Mikrokernen, was auf eine mögliche Rolle der CES-vermittelten Entgiftung bei der Genotoxizität von Lasiocarpin hindeutet. Auch intrazelluläres Glutathion (GSH) spielt eine wichtige Rolle bei der Entgiftung von Xenobiotika oder Toxinen. Zellen, die mit L-Buthioninsulfoximin (BSO) vorbehandelt worden waren, um den GSH-Gehalt zu reduzieren, waren signifikant empfindlicher für die Induktion der Mikronukleusbildung durch Lasiocarpin, was die Bedeutung von GSH für die PA-induzierte Genotoxizität zeigt.
Chinidin (Q) und Nelfinavir (NFR) sind OCT1- bzw. OATP1B1-Influx-Transporter-Inhibitoren, die die Mikronukleus-Induktion durch Lasiocarpin reduzierten (nur Chinidin signifikant), aber nicht vollständig, was auf eine Relevanz von OCT1 für die PA-Aufnahme in HepG2-Zellen hindeutet.Verapamil (V) und Benzbromaron (Bz) sind MDR1- bzw. MRP2-Efflux-Transporter-Inhibitoren und verursachten eine leicht erhöhte Mikronukleus-Induktion durch Lasiocarpin (signifikant nur für Benzbromaron), was die Rolle von Efflux-Transportern bei der PA-induzierten Genotoxizität aufzeigt.
Der Mechanismus der PA-induzierten Genotoxizität wurde auf der Grundlage von oxidativem Stress durch die Bildung von reaktiven Sauerstoffspezies (ROS) in HepG2-Zellen weiter untersucht. Eine Überproduktion von ROS kann zelluläre Makromoleküle wie DNA vernetzen, was zu genomischen Schäden führt. Eine äquimolare Konzentration von 10 μM von Lasiocarpin (Open-Diester PA), Riddelliin (Cyclic-Diester PA) und Europin (Monoester) induzierte signifikant die ROS-Produktion, wobei die höchste ROS-Erzeugung nach Lasiocarpin-Behandlung beobachtet wurde, gefolgt von Riddelliin und Europin. Mit Lycopsamin (10 μM; Monoester PA) wurde auch bei höherer Konzentration (320 μM) keine signifikante Steigerung der ROS-Produktion gefunden.
Um die Beteiligung von ROS am Mechanismus der Genotoxizität einzelner PAs genauer zu betrachten und die bisherigen Ergebnisse zu bestätigen, wurden weitere Untersuchungen in Anwesenheit des Sauerstoffradikalfängers N-Acetylcysteine (NAC) in Kombination mit Lasiocarpin, Riddelliin oder Europin durchgeführt. Diese Analyse ergab eine signifikante Abnahme der ROS-Produktion nach der Kombination von NAC mit Lasiocarpin, Riddelliin und Europin. Darüber hinaus induzierten Lasiocarpin, Riddelliin und Europin Veränderungen im mitochondrialen Membranpotenzial. Dies deutet darauf hin, dass ROS vermehrt in den Mitochondrien der Zellen gebildet werden.
Aus in vivo Daten ist bekannt, dass hepatische sinusoidale Epithelzellen (HSECs) die Zelltypen innerhalb der Leber sind, die nach der metabolischen Aktivierung von PAs zuerst geschädigt werden. Jedoch exprimieren HSECs nicht die erforderlichen Stoffwechselenzyme für die Aktivierung von PAs. Um diese Situation nachzuahmen, wurden HepG2-Zellen verwendet, um PAs in einer Kokultur mit HeLa H2B-GFP-Zellen als nicht-metabolisch aktive Nachbarn metabolisch zu aktivieren. Durch die grün fluoreszierende GFP-Markierung konnten die HeLa-Zellen in der Co-Kultur leicht identifiziert werden. Die PAs Europine, Riddelliin und Lasiocarpin induzierten die Bildung von Mikrokernen in HepG2-Zellen und in HeLa H2B-GFP-Zellen, die mit HepG2-Zellen kokultiviert wurden, jedoch nicht in HeLa H2B-GFP-Zellen, die allein kultiviert wurden. Die metabolische Hemmung von CYP 450-Enzymen mit Ketoconazol hob die Mikronukleusbildung, welche durch die zuvor getesteten PAs induziert wurde, auf. Die Efflux-Transporter-Inhibitoren Verapamil und Benzbromaron reduzierten die Mikronukleusbildung in der Kokultur. Darüber hinaus wurden mitotische Störungen als zusätzlicher genotoxischer Wirkmechanismus in der Co-Kultur aus HepG2-Zellen und in HeLa H2B-GFP-Zellen beobachtet, jedoch nicht in HeLa H2B-GFP-Zellen, die allein kultiviert wurden. Zusammengefasst deuten diese Ergebnisse darauf hin, dass PAs durch HepG2-Zellen bioaktiviert werden können und aus PAs gebildete Metabolite genomische Schäden in kokultivierten, nicht-metabolisch aktiven HeLa-Zellen induzierten.

Abschließend zeigen Kombinationen der PAs Lasiocarpin und Riddelliin sowohl in HepG2-Zellen als auch in der Co-Kultur eher einen additiven Effekt als einen Synergismus. Diese Studie liefert daher Unterstützung für die Annahme, dass die Dosisaddition zur Charakterisierung des genotoxischen Risikos von in einem Gemisch vorhandenen PAs angewendet werden kann.
KW  - Pyrrolizidine alkaloids
KW  - HeLa H2B-GFP-Zellen
KW  - Pyrrolizidinalkaloide
KW  - Kleinkern
KW  - Mutagenität
KW  - Genotoxizität
KW  - DNA-Vernetzung
KW  - mitotische Störung
KW  - Co-culture
KW  - metabolische Aktivierung
KW  - Membrantransporter
KW  - metabolische Enzyme
KW  - HepG2-Zellen
KW  - Genotoxicity
KW  - Micronuclei
KW  - DNA crosslink
KW  - Mitotic disturbance
KW  - Metabolic activation
KW  - Membrane transporters
KW  - Metabolic enzymes
KW  - HepG2 cells
KW  - HeLa H2B-GFP cells
KW  - micronucleus
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370376
ER  - 
TY  - THES
A1  - Choi, Jihyoung
T1  - Development of an Add-On Electrode for Non-Invasive Monitoring in Bioreactor Cultures and Medical Devices
T1  - Entwicklung einer Zusatzelektrode für das nicht-invasive Monitoring von Bioreaktorkulturen und Medizinprodukten
N2  - Electrochemical impedance spectroscopy (EIS) is a valuable technique analyzing electrochemical behavior of biological systems such as electrical characterization of cells and biomolecules, drug screening, and biomaterials in biomedical field. In EIS, an alternating current (AC) power signal is applied to the biological system, and the impedance of the system is measured over a range of frequencies. 
In vitro culture models of endothelial or epithelial barrier tissue can be achieved by culturing barrier tissue on scaffolds made with synthetic or biological materials that provide separate compartments (apical and basal sides), allowing for further studies on drug transport. EIS is a great candidate for non-invasive and real-time monitoring of the electrical properties that correlate with barrier integrity during the tissue modeling. Although commercially available transendothelial/transepithelial electrical resistance (TEER) measurement devices are widely used, their use is particularly common in static transwell culture. EIS is considered more suitable than TEER measurement devices in bioreactor cultures that involve dynamic fluid flow to obtain accurate and reliable measurements. Furthermore, while TEER measurement devices can only assess resistance at a single frequency, EIS measurements can capture both resistance and capacitance properties of cells, providing additional information about the cellular barrier's characteristics across various frequencies. Incorporating EIS into a bioreactor system requires the careful optimization of electrode integration within the bioreactor setup and measurement parameters to ensure accurate EIS measurements. Since bioreactors vary in size and design depending on the purpose of the study, most studies have reported using an electrode system specifically designed for a particular bioreactor. The aim of this work was to produce multi-applicable electrodes and established methods for automated non-invasive and real-time monitoring using the EIS technique in bioreactor cultures. Key to the electrode material, titanium nitride (TiN) coating was fabricated on different substrates (materials and shape) using physical vapor deposition (PVD) and housed in a polydimethylsiloxane (PDMS) structure to allow the electrodes to function as independent units. Various electrode designs were evaluated for double-layer capacitance and morphology using EIS and scanning electron microscopy (SEM), respectively. The TiN-coated tube electrode was identified as the optimal choice. Furthermore, EIS measurements were performed to examine the impact of influential parameters related to culture conditions on the TiN-coated electrode system. In order to demonstrate the versatility of the electrodes, these electrodes were then integrated into in different types of perfusion bioreactors for monitoring barrier cells.  Blood-brain barrier (BBB) cells were cultured in the newly developed dynamic flow bioreactor, while human umblical vascular endothelial cells (HUVECs) and Caco-2 cells were cultured in the miniature hollow fiber bioreactor (HFBR). As a result, the TiN-coated tube electrode system enabled investigation of BBB barrier integrity in long-term bioreactor culture. While EIS measurement could not detect HUVECs electrical properties in miniature HFBR culture, there was the possibility of measuring the barrier integrity of Caco-2 cells, indicating potential usefulness for evaluating their barrier function. Following the bioreactor cultures, the application of the TiN-coated tube electrode was expanded to hemofiltration, based on the hypothesis that the EIS system may be used to monitor clotting or clogging phenomena in hemofiltration. The findings suggest that the EIS monitoring system can track changes in ion concentration of blood before and after hemofiltration in real-time, which may serve as an indicator of clogging of filter membranes. Overall, our research demonstrates the potential of TiN-coated tube electrodes for sensitive and versatile non-invasive monitoring in bioreactor cultures and medical devices.
N2  - Die elektrochemische Impedanzspektroskopie (EIS) ist eine nützliche Methode, um das elektrochemische Verhalten von biologischen Systemen zu analysieren, wie z.B. die elektrische Charakterisierung von Zellen und Biomolekülen, Drug Screening und Biomaterialien im biomedizinischen Bereich. Für die EIS wird ein Wechselstrom an das biologische System angeschlossen und die Impedanz des Systems über einen Frequenzbereich gemessen. 
In vitro-Modelle von Gewebekulturen epithelialer Barrieren können mithilfe künstlicher oder biologischer Materialien, die über unterschiedliche Kompartimente (apikale und basolaterale Seite) verfügen, hergestellt werden und ermöglichen weitere Untersuchungen zum Transport von Arzneistoffen. Die EIS bietet dabei eine hervorragende Methode für das nicht-invasive Echtzeit-Monitoring der elektrischen Eigenschaften, die mit der Barriere-Integrität während der Gewebeentwicklung korreliert. Obwohl kommerziell erhältliche Geräte zur Messung des transendothelialen/transepithelialen elektrischen Widerstands (TEER) umfangreich verwendet werden, ist ihre Verwendung besonders bei statischen Transwell-Kulturen verbreitet. Durch die EIS kann im Gegensatz zur TEER-Messung für Bioreaktor-Kulturen, die einen dynamischen Medienfluss aufweisen, genauere und verlässliche Messungen erhalten werden. Zudem können EIS-Messungen anders als die TEER-Messung, die nur den Widerstand einer einzelnen Frequenz misst, gleichzeitig den elektrischen Widerstand und die Kapazität von Zellen erfassen und damit zusätzliche Informationen über die zellulären Barriereeigenschaften über verschiedene Frequenzen hinweg liefern. Der EIS-Einbau in ein Bioreaktor-System bedarf einer sorgfältigen Optimierung der Elektrodenintegration in das Bioreaktor-Setup und der Messparameter, um akkurate EIS-Messungen durchführen zu können. Da Bioreaktoren abhängig vom Untersuchungszweck in ihrer Größe und ihrem Design variieren, verwenden die meisten Studien speziell entwickelte Elektrodensysteme für einzelne Bioreaktoren. Das Ziel dieser Arbeit war die Herstellung von vielseitig anwendbaren Elektroden und etablierten Methoden für das automatisierte nicht-invasive Echtzeit-Monitoring von Bioreaktor-Kulturen mithilfe der EIS. Entscheidend für das Elektrodenmaterial war die Titannitrid (TiN)-Beschichtung, die auf verschiedenen Substraten (Materialien und Formen) durch Physical Vapor Deposition (PVD) hergestellt und in einer Polydimethylsiloxan (PDMS)-Struktur untergebracht wurde, damit die Elektroden unabhängig voneinander arbeiten können. Verschiedene Elektrodendesigns wurden auf Doppelschicht-Kapazität mithilfe der EIS bzw. auf die Morphologie mit Rasterelektronenmikroskopie untersucht. Die TiN-beschichteten Elektroden in Röhrenform erwiesen sich als optimal. Weiterhin wurden EIS-Messungen durchgeführt, um die Auswirkung von beeinflussenden Parametern auf die Kulturbedingungen durch das TiN-beschichtete Elektrodensystem zu untersuchen. Um die Vielseitigkeit der Elektroden aufzuzeigen, wurden diese anschließend zum Monitoring von Barriere-bildenden Zellen in unterschiedliche Perfusionsbioreaktoren integriert. Zellen der Blut-Hirn-Schranke (BHS) wurden im neu entwickelten dynamischen Flussreaktor kultiviert, wohingegen humane umbilikale vaskuläre Endothelzellen (HUVEC) und Caco-2-Zellen in Hohlfaserbioreaktoren (HFBR) in Miniaturform kultiviert wurden. Das TiN-beschichtete Röhrenelektrodensystem ermöglichte die Untersuchung der BHS-Barrieren-Integrität in einer Langzeit-Bioreaktorkultur. Während die EIS-Messung in der Miniaturform-HFBR-Kultur keine elektrischen Eigenschaften der HUVECs detektieren konnte, war es möglich, eine Barriere-Integrität der Caco-2-Zellen zu messen, die den potentiellen Nutzen für die Evaluierung deren Barrierefunktion aufzeigt. Nach den Bioreaktorkulturen wurde die Anwendung der TiN-beschichteten Röhrenelektrode auf die Hämofiltration erweitert, auf Grundlage der Hypothese, dass das EIS-System ein Gerinnen oder Verstopfen während der Hämofiltration überwachen könnte. Die Ergebnisse zeigen, dass das EIS-Monitoring-System Veränderungen in der Ionenkonzentration des Blutes vor und nach Hämofiltration in Echtzeit verfolgen kann, welches eventuell als Messgröße für ein Verstopfen der Filtermembranen genutzt werden kann. Insgesamt weisen TiN-beschichtete Röhrenelektroden unseren Forschungen zufolge ein großes Potential für ein empfindliches und vielfältiges nicht-invasives Monitoring von Bioreaktorkulturen und Medizingeräte auf.
KW  - Monitoring
KW  - Tissue Engineering
KW  - Electrode
KW  - Perfusion Bioreactor
KW  - Hemofiltration
KW  - Medizinprodukt
KW  - Electrochemical Impedance Spectroscopy
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358232
ER  - 
TY  - THES
A1  - Kutschka, Ilona
T1  - Activation of the integrated stress response induces remodeling of cardiac metabolism in Barth Syndrome
T1  - Aktivierung der "Integrated Stress Response" führt zur Umstellung des kardialen Metabolismus im Barth Syndrom
N2  - Barth Syndrome (BTHS) is an inherited X-chromosomal linked disorder, characterized by early development of cardiomyopathy, immune system defects, skeletal muscle myopathy and growth retardation. The disease displays a wide variety of symptoms including heart failure, exercise intolerance and fatigue due to the muscle weakness. The cause of the disease are mutations in the gene encoding for the mitochondrial transacylase Tafazzin (TAZ), which is important for remodeling of the phospholipid cardiolipin (CL). All mutations result in a pronounced decrease of the functional enzyme leading to an increase of monolysocardiolipin (MLCL), the precursor of mature CL, and a decrease in mature CL itself. CL is a hallmark phospholipid of mitochondrial membranes, highly enriched in the inner mitochondrial membrane (IMM). It is not only important for the formation of the cristae structures, but also for the function of different protein complexes associated with the mitochondrial membrane. Reduced levels of mature CL cause remodeling of the respiratory chain supercomplexes, impaired respiration, defects in the Krebs cycle and a loss of mitochondrial calcium uniporter (MCU) protein. The defective Ca2+ handling causes impaired redox homeostasis and energy metabolism resulting in cellular arrhythmias and defective electrical conduction. In an uncompensated situation, blunting mitochondrial Ca2+ uptake provokes increased mitochondrial emission of H2O2 during workload transitions, related to oxidation of NADPH, which is required to regenerate anti-oxidative enzymes. However, in the hearts and cardiac myocytes of mice with a global knock-down of the Taz gene (Taz-KD), no increase in mitochondrial ROS was observed, suggesting that other metabolic pathways may have compensated for reduced Krebs cycle activation.
The healthy heart produces most of its energy by consuming fatty acids. In this study, the fatty acid uptake into mitochondria and their further degradation was investigated, which showed a switch of the metabolism in general in the Taz-KD mouse model. In vivo studies revealed an increase of glucose uptake into the heart and decreased fatty acid uptake and oxidation. Disturbed energy conversion resulted in activation of retrograde signaling pathways, implicating overall changes in the cell metabolism. Upregulated integrated stress response (ISR) was confirmed by increased levels of the downstream target, i.e., the activating transcription factor 4 (ATF4). A Tafazzin knockout mouse embryonal fibroblast cell model (TazKO) was used to inhibit the ISR using siRNA transfection or pharmaceutical inhibition. This verified the central role of
II
the ISR in regulating the metabolism in BTHS. Moreover, an increased metabolic flux into glutathione biosynthesis was observed, which supports redox homeostasis. In vivo PET-CT scans depicted elevated activity of the xCT system in the BTHS mouse heart, which transports essential amino acids for the biosynthesis of glutathione precursors. Furthermore, the stress induced signaling pathway also affected the glutamate metabolism, which fuels into the Krebs cycle via -ketoglutarate and therefore supports energy converting pathways. In summary, this thesis provides novel insights into the energy metabolism and redox homeostasis in Barth syndrome cardiomyopathy and its regulation by the integrated stress response, which plays a central role in the metabolic alterations. The aim of the thesis was to improve the understanding of these metabolic changes and to identify novel targets, which can provide new possibilities for therapeutic intervention in Barth syndrome.
N2  - Barth Syndrome (BTHS) ist eine X-chromosomal vererbbare Erkrankung, welche sich in der frühen Entstehung von Kardiomyopathie, Störungen des Immunsystems, Skelettmuskelschwäche und Wachstumsverzögerungen manifestiert. Das Krankheitsbild ist sehr variabel mit milden Symptomen bis hin zu sehr schwerwiegenden Fällen, bei denen die schnelle Verschlechterung der Kardiomyopathie bereits in jungen Jahren eine Herztransplantation erfordern kann. Betroffenen Patienten zeigen eine deutliche Intoleranz gegenüber körperlicher Anstrengung, welche mit schneller Müdigkeit einhergeht. Die Krankheit wird durch verschiedene Mutationen auf dem Gen für die mitochondriale Transacylase Tafazzin (TAZ) ausgelöst. Die Mutationen führen zu einem Funktionsverlust des Enzyms, welches in der Biosynthese des Phospholipids Cardiolipin (CL) eine entscheidende Rolle spielt. Die Vorstufe des Lipids, das sogenannte Monolysocardiolipin (MLCL), reichert sich dadurch an, wohingegen die Menge an reifem CL entscheidend verringert ist. CL ist ein bedeutendes Phospholipid in den Mitochondrien, wo es vor allem in der inneren Mitochondrien Membran vorkommt. CL ist einerseits wichtig für die Ausbildung der Cristae Strukturen der inneren Mitochondrien Membran. Darüber hinaus ist es notwendig für die Struktur und Funktion verschiedenster Proteinkomplexe in der Membran, welche dadurch erst ihre volle Funktionsfähigkeit erhalten. Es wurde bereits gezeigt, dass der Verlust von reifem CL in BTHS zu einer Dissoziation der Superkomplexe der Atmungskette führt, welche dadurch in ihrer Funktion beeinträchtigt ist. Zusätzlich sind Störungen im Krebs Zyklus und der Kalziumaufnahme durch den mitochondriellen Kalzium (Ca2+) -Uniporter (MCU) Komplex bekannt. Die beeinträchtigte mitochondriale Ca2+ Aufnahme beeinflusst sowohl die Redox Homöostase als auch den Energie Metabolismus, was zu Arrhythmien und einer Störung der elektrischen Weiterleitung im Herzen führt. Im gesunden Herzen gewinnen die Herzmuskelzellen den Hauptanteil ihrer Energie aus dem Abbau von Fettsäuren. In dieser Studie wurde durch die Untersuchung des Fettsäurestoffwechsels im Taz knockdown Mausmodell (Taz-KD) gezeigt, dass eine deutliche Reduktion in Proteinen vorliegt, welche für die Aufnahme und die Verstoffwechselung der Fettsäuren in den Mitochondrien verantwortlich sind. Diese Veränderungen führten in vivo zu einer verringerten Aufnahme und Verstoffwechselung von Fettsäuren und einer Erhöhten Aufnahme von Glucose. Dysfunktionale Mitochondrien aktivieren retrograde Signalwege, welche eine generelle
IV
Umstellung des Metabolismus zur Folge haben. Eine erhöhte Menge des Transkriptionsfaktors ATF4, welcher sowohl Fettsäure- als auch Aminosäuremetabolismus beeinflusst, zeigte die Aktivierung der sogenannten „Integrated stress response“ (ISR). Ein Zellmodel embryonaler Fibroblasten aus der Maus mit einem Taz knockout (TazKO) wurde verwendet um die ISR durch siRNA Transfektion oder einem pharmakologischen Inhibitor zu blockieren. Dadurch konnte die zentrale Rolle der ISR in der Umstellung des Metabolismus bestätigt werden. Zusätzlich konnte eine erhöhte metabolische Aktivität in Richtung der Glutathion Biosynthese beobachtet werden, welche für die Redox Homöostase in den Mitochondrien von Bedeutung ist. In vivo PET-CT Untersuchungen zeigten eine erhöhte Aktivität des xCT Systems im Herzen des BTHS Mausmodells auf. Dies dient der Aufnahme von Aminosäuren, welche für die Glutathion Biosynthese benötigt werden. Hinzu kommt, dass die Aktivierung des Stresssignalweges den Glutamat Stoffwechsel in der Zelle beeinflusste. Über -Ketoglutarat trägt Glutamat so vermehrt zur Energiegewinnung bei. Das Ziel dieser Doktorarbeit war es, die metabolischen Veränderungen in BTHS zu untersuchen, um die veränderten Vorgänge besser zu verstehen und so neue mögliche Angriffspunkte für Therapiemöglichkeiten zu identifizieren.
KW  - Herzmuskelkrankheit
KW  - Mitochondrium
KW  - Stoffwechsel
KW  - Barth Syndrome
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358186
ER  - 
TY  - THES
A1  - Wilhelmi, Kai Alexander
T1  - Untersuchung von Veränderungen der myelinisierten Nervenfasern durch Entmarkung in Haut- und Nervenbiopsien von Patienten mit Polyneuropathie
T1  - Examination of changes in myelinated nerve fibers due to demyelination in skin and nerve biopsies of patients with polyneuropathy
N2  - In dieser Arbeit wurde durch das immunhistochemische Anfärben von nodalen (Natriumkanäle, NF), paranodalen (Caspr, NF) und internodalen (MBP) Proteinen der in Fingerhautbiopsien vorhanden Nervenfasern untersucht, ob eine Veränderung der typischen Verteilungsmuster dieser Proteine, eine demyelinisierende Polyneuropathie anzeigen kann. Dazu wurden am Universitätsklinikum Würzburg prospektiv 93 Polyneuropathie-Patienten und 25 Kontrollpersonen rekrutiert. Bei allen Patienten wurden Hautstanzbiospien am Zeigefinger durchgeführt. Bei 35 Patienten mit schweren oder unklaren Verläufen, wurden konsiliarisch Nervus suralis Biopsien durchgeführt. Aus einem Abschnitt von 27 dieser Biopsien, konnten im Rahmen dieser Arbeit Zupfnervenpräparate angefertigt und analog zu den Hautbiopsien ausgewertet werden. Aus der Routinediagnostik der Klinik flossen weiterhin die Ergebnisse der elektrophysiologischen Routinediagnostik und der Histologiebefund der Nervus suralis Biopsien in die Auswertung ein. 
Zusammenfassend kamen veränderte Natriumkanalbanden in Fingerhautbiopsien signifikant häufiger bei Patienten mit elektrophysiologisch als demyelinisierend befundeten Polyneuropathien, als bei Patienten mit elektrophysiologisch als axonal befundeten Polyneuropathien vor. Vielfach fanden sich veränderte Natriumkanalbanden inmitten para- und internodal unauffälliger Schnürringe und umgekehrt. Diese Beobachtung stützt die bereits in Vorarbeiten vorgeschlagene und in der aktuellen Leitlinie zur Diagnostik für Polyneuropathien aufgegriffene Entität der Paranodopathien (Uncini, Susuki, & Yuki, 2013). Möglich wäre, dass eine veränderte Verteilung der Natriumkanäle die schnelle Leitfähigkeit beeinträchtigen und somit trotz intakter Bemarkung, elektrophysiologisch das Bild einer demyelinisierenden Neuropathie vermittelt. Ein direkter Zusammenhang zwischen dem Auftreten von doppelten und verlängerten Natriumkanalbanden und einzelnen Messwerten (z.B. Amplituden und Latenzzeiten) fand sich nicht. Auch in den Zupfnervenpräparaten der Nervus suralis Biopsien, konnten o.g. Verteilungsmuster untersucht werden. Deren Vorkommen zeigte sich als unabhängig vom elektrophysiologischen und histologischen Befund, von der Ätiologie der PNP und von den gefundenen Veränderungen in den Hautbiopsien des betreffenden Patienten.
N2  - Myelinated nerve fibers in finger skin biopsies and sural nerve biopsies were examined using immunohistochemical staining to detect changes in the typical distribution patterns of nodal (voltage-gated sodium channels, neurofascin 186), paranodal (Caspr, neurofascin 155), and internodal (myelin basic protein) proteins, aiming to identify indicators for demyelinating polyneuropathies. A total of 93 polyneuropathy patients and 25 control subjects were prospectively recruited from the University Hospital Würzburg. Skin punch biopsies were conducted on all patients and control subjects. Additionally, sural nerve biopsies were performed on a consultative basis for 35 patients. Teased nerve fiber preparations were made from a section of 27 of these biopsies and evaluated similarly to the skin biopsies. In summary, altered sodium channel bands in myelinated nerve fibers from finger skin biopsies were significantly more prevalent in patients electrophysiologically diagnosed with demyelinating polyneuropathies. However, there was no significant difference in the means of individual electrophysiological measurements between patients with and without changes in the immunohistochemical stainings. Each of the investigated changes was significantly more common in the polyneuropathy group than in the control group. Further correlations, particularly in the comparison of results from skin and sural nerve biopsies, were not found
KW  - Polyneuropathie
KW  - Ranvier-Schnürring
KW  - Entmarkung
KW  - Elektrophysiologie
KW  - Caspr
KW  - Neurofascin
KW  - MBP
KW  - spannungsgesteuerter Natriumkanal
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360046
ER  - 
TY  - JOUR
A1  - Gerlich, C.
A1  - Andreica, I.
A1  - Küffner, R.
A1  - Krause, D.
A1  - Lakomek, H. J.
A1  - Reusch, A.
A1  - Braun, J.
T1  - Evaluation einer Basisschulung für Patienten mit rheumatoider Arthritis
T1  - Evaluation of a basic educational program for patients with rheumatoid arthritis
JF  - Zeitschrift für Rheumatologie
N2  - Hintergrund
Ein neues Rahmenkonzept hat die flexible Ableitung und Nutzung von rheumatologischen Schulungsprogrammen für unterschiedliche Versorgungsbereiche ermöglicht. Auf dieser Grundlage wurde eine 5‑stündige Basisschulung für Patienten mit rheumatoider Arthritis (RA) entwickelt, es wurden rheumatologische Fachärzte und Psychologen trainiert, und dann wurde die Wirksamkeit nach dem Wirkmodell der Patientenschulung evaluiert.

Methoden
Mit dem Studiendesign einer extern randomisierten Wartekontrollgruppenstudie mit 3 Messzeitpunkten wurde geprüft, wie sich die 5‑stündige Basisschulung auf das Erkrankungs- und Behandlungswissen sowie auf die Gesundheitskompetenz von RA-Patienten (n = 249) auswirkt. Weitere Fragen betrafen Einstellungsparameter, Kommunikationskompetenz, Erkrankungsauswirkungen und die Zufriedenheit mit der Schulung. Die Auswertungen erfolgten auf Intention-to-treat-Basis mit Kovarianzanalysen für die Hauptzielgrößen unter Berücksichtigung des Ausgangswertes.

Ergebnisse
Die Analysen zeigen, dass die Basisschulung RA wirksam ist. Noch 3 Monate nach der Schulung verfügten die Schulungsteilnehmer über mehr Wissen und Gesundheitskompetenz als die Wartekontrollgruppe mit kleinem bis mittelgroßem Effekt (d = 0,37 bzw. 0,38). In den Nebenzielgrößen zeigten sich mit Ausnahme der Krankheitskommunikation keine weiteren Schulungseffekte.

Diskussion
Die Basisschulung bietet eine gute Grundlage, auf der weitere Interventionen zur Verbesserung von Einstellungs- und Erkrankungsparametern aufbauen können. Sie eignet sich damit als zentraler Baustein für die rheumatologische Versorgung auf verschiedenen Ebenen.
N2  - Background
A new conceptual framework has enabled the flexible development of rheumatological patient educational programs for different healthcare settings. On this basis, a 5‑h basic training program for patients with rheumatoid arthritis (RA) was developed to be used in specialized centers. Rheumatologists and psychologists were first trained and then the efficacy of the patient training program was evaluated based on the causal model of patient education.

Methods
The externally randomized waiting control group study with 249 RA patients included 3 measurement points. The impact of the 5‑h basic training on disease and treatment-related knowledge as well as health competence of RA patients was examined. Secondary questions included attitudinal parameters, communication competence, effects on the disease and satisfaction with the educational program. Data were analyzed on an intention to treat basis by means of covariance analyses for the main target variables, adjusted for baseline values.

Results
The analyses showed that the training program was effective. Even 3 months after training, participants reported more knowledge and health competence than the waiting control group, with small to medium-sized effects (d = 0.37 and 0.38, respectively). With the exception of disease communication, no other effects of training were observed in the secondary objectives.

Conclusion
The basic training program provides a good foundation to develop further interventions to improve attitudinal and disease parameters. It can serve as a central component for rheumatological healthcare for patients with RA at various levels.
KW  - Rheumatoide Arthritis
KW  - Patientenschulung
KW  - Evaluation
KW  - Rheumatoid arthritis
KW  - Patient education
KW  - Evaluation
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-280359
VL  - 79
ER  - 
TY  - JOUR
A1  - Hauer, Nadine N.
A1  - Popp, Bernt
A1  - Taher, Leila
A1  - Vogl, Carina
A1  - Dhandapany, Perundurai S.
A1  - Büttner, Christian
A1  - Uebe, Steffen
A1  - Sticht, Heinrich
A1  - Ferrazzi, Fulvia
A1  - Ekici, Arif B.
A1  - De Luca, Alessandro
A1  - Klinger, Patrizia
A1  - Kraus, Cornelia
A1  - Zweier, Christiane
A1  - Wiesener, Antje
A1  - Abou Jamra, Rami
A1  - Kunstmann, Erdmute
A1  - Rauch, Anita
A1  - Wieczorek, Dagmar
A1  - Jung, Anna-Marie
A1  - Rohrer, Tilman R.
A1  - Zenker, Martin
A1  - Doerr, Helmuth-Guenther
A1  - Reis, André
A1  - Thiel, Christian T.
T1  - Evolutionary conserved networks of human height identify multiple Mendelian causes of short stature
JF  - European Journal of Human Genetics
N2  - Height is a heritable and highly heterogeneous trait. Short stature affects 3% of the population and in most cases is genetic in origin. After excluding known causes, 67% of affected individuals remain without diagnosis. To identify novel candidate genes for short stature, we performed exome sequencing in 254 unrelated families with short stature of unknown cause and identified variants in 63 candidate genes in 92 (36%) independent families. Based on systematic characterization of variants and functional analysis including expression in chondrocytes, we classified 13 genes as strong candidates. Whereas variants in at least two families were detected for all 13 candidates, two genes had variants in 6 (UBR4) and 8 (LAMA5) families, respectively. To facilitate their characterization, we established a clustered network of 1025 known growth and short stature genes, which yielded 29 significantly enriched clusters, including skeletal system development, appendage development, metabolic processes, and ciliopathy. Eleven of the candidate genes mapped to 21 of these clusters, including CPZ, EDEM3, FBRS, IFT81, KCND1, PLXNA3, RASA3, SLC7A8, UBR4, USP45, and ZFHX3. Fifty additional growth-related candidates we identified await confirmation in other affected families. Our study identifies Mendelian forms of growth retardation as an important component of idiopathic short stature.
KW  - disease genetics
KW  - DNA sequencing
KW  - genetic counselling
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227899
VL  - 27
ER  - 
TY  - JOUR
A1  - Gilder, Stuart A.
A1  - Wack, Michael
A1  - Kaub, Leon
A1  - Roud, Sophie C.
A1  - Petersen, Nikolai
A1  - Heinsen, Helmut
A1  - Hillenbrand, Peter
A1  - Milz, Stefan
A1  - Schmitz, Chistoph
T1  - Distribution of magnetic remanence carriers in the human brain
JF  - Scientific Reports
N2  - That the human brain contains magnetite is well established; however, its spatial distribution in the brain has remained unknown. We present room temperature, remanent magnetization measurements on 822 specimens from seven dissected whole human brains in order to systematically map concentrations of magnetic remanence carriers. Median saturation remanent magnetizations from the cerebellum were approximately twice as high as those from the cerebral cortex in all seven cases (statistically significantly distinct, p = 0.016). Brain stems were over two times higher in magnetization on average than the cerebral cortex. The ventral (lowermost) horizontal layer of the cerebral cortex was consistently more magnetic than the average cerebral cortex in each of the seven studied cases. Although exceptions existed, the reproducible magnetization patterns lead us to conclude that magnetite is preferentially partitioned in the human brain, specifically in the cerebellum and brain stem.
KW  - brain
KW  - neurophysiology
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233035
VL  - 8
ER  - 
TY  - JOUR
A1  - Gotru, Sanjeev Kiran
A1  - van Geffen, Johanna P.
A1  - Nagy, Magdolna
A1  - Mammadova-Bach, Elmina
A1  - Eilenberger, Julia
A1  - Volz, Julia
A1  - Manukjan, Georgi
A1  - Schulze, Harald
A1  - Wagner, Leonard
A1  - Eber, Stefan
A1  - Schambeck, Christian
A1  - Deppermann, Carsten
A1  - Brouns, Sanne
A1  - Nurden, Paquita
A1  - Greinacher, Andreas
A1  - Sachs, Ulrich
A1  - Nieswandt, Bernhard
A1  - Hermanns, Heike M.
A1  - Heemskerk, Johan W. M.
A1  - Braun, Attila
T1  - Defective Zn2+ homeostasis in mouse and human platelets with α- and δ-storage pool diseases
JF  - Scientific Reports
N2  - Zinc (Zn2+) can modulate platelet and coagulation activation pathways, including fibrin formation. Here, we studied the (patho)physiological consequences of abnormal platelet Zn2+ storage and release. To visualize Zn2+ storage in human and mouse platelets, the Zn2+ specific fluorescent dye FluoZin3 was used. In resting platelets, the dye transiently accumulated into distinct cytosolic puncta, which were lost upon platelet activation. Platelets isolated from Unc13d−/− mice, characterized by combined defects of α/δ granular release, showed a markedly impaired Zn2+ release upon activation. Platelets from Nbeal2−/− mice mimicking Gray platelet syndrome (GPS), characterized by primarily loss of the α-granule content, had strongly reduced Zn2+ levels, which was also confirmed in primary megakaryocytes. In human platelets isolated from patients with GPS, Hermansky-Pudlak Syndrome (HPS) and Storage Pool Disease (SPD) altered Zn2+ homeostasis was detected. In turbidity and flow based assays, platelet-dependent fibrin formation was impaired in both Nbeal2−/− and Unc13d−/− mice, and the impairment could be partially restored by extracellular Zn2+. Altogether, we conclude that the release of ionic Zn2+ store from secretory granules upon platelet activation contributes to the procoagulant role of Zn2+ in platelet-dependent fibrin formation.
KW  - coagulation system
KW  - metals
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227455
VL  - 9
ER  - 
TY  - JOUR
A1  - Gore, Lia
A1  - Locatelli, Franco
A1  - Zugmaier, Gerhard
A1  - Handgretinger, Rupert
A1  - O'Brien, Maureen M.
A1  - Bader, Peter
A1  - Bhojwani, Deepa
A1  - Schlegel, Paul-Gerhardt
A1  - Tuglus, Catherine A.
A1  - Stackelberg, Arend von
T1  - Survival after blinatumomab treatment in pediatric patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia
JF  - Blood Cancer Journal
N2  - no abstract available
KW  - acute lymphocytic leukaemia
KW  - immunotherapy
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230726
VL  - 8
ER  - 
TY  - JOUR
A1  - Franchini, Paolo
A1  - Jones, Julia C.
A1  - Xiong, Peiwen
A1  - Kneitz, Susanne
A1  - Gompert, Zachariah
A1  - Warren, Wesley C.
A1  - Walter, Ronald B.
A1  - Meyer, Axel
A1  - Schartl, Manfred
T1  - Long-term experimental hybridisation results in the evolution of a new sex chromosome in swordtail fish
JF  - Nature Communications
N2  - The remarkable diversity of sex determination mechanisms known in fish may be fuelled by exceptionally high rates of sex chromosome turnovers or transitions. However, the evolutionary causes and genomic mechanisms underlying this variation and instability are yet to be understood. Here we report on an over 30-year evolutionary experiment in which we tested the genomic consequences of hybridisation and selection between two Xiphophorus fish species with different sex chromosome systems. We find that introgression and imposing selection for pigmentation phenotypes results in the retention of an unexpectedly large maternally derived genomic region. During the hybridisation process, the sex-determining region of the X chromosome from one parental species was translocated to an autosome in the hybrids leading to the evolution of a new sex chromosome. Our results highlight the complexity of factors contributing to patterns observed in hybrid genomes, and we experimentally demonstrate that hybridisation can catalyze rapid evolution of a new sex chromosome.
KW  - evolutionary genetics
KW  - experimental evolution
KW  - genome evolution
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228396
VL  - 9
ER  - 
TY  - THES
A1  - Stürzebecher, Paulina Elena
T1  - Die Rolle von LASP1 in der Pathogenese der Atherosklerose im murinen Modell
T1  - The role of LASP1 in the pathogenesis of atherosclerosis in mice
N2  - Das regulatorische Gerüst-Protein LASP1, welches aus der Krebsforschung bekannt ist, wurde 2012 in humanen Makrophagen, den Protagonisten der Atherosklerose nachgewiesen. LASP1 ist durch seine Lokalisation an dynamischen Aktinskelettkonstruktionen (vgl. Invadopodien, Podosomen), nachweislich an Zellmigration, Proliferation und Invasionsfähigkeit bestimmter Tumorzellen beteiligt. Aufgrund einer großen Schnittmenge der Entstehungsmechanismen und zugrundeliegenden Signalwegen von Krebserkrankungen und Atherosklerose wurde LASP1 im Zusammenhang der Atherosklerose untersucht. In einem 16 Wochen Hochfettdiätversuch zeigten LASP1.Ldlr-/--Mäuse mehr atherosklerotische Läsionen in der Gesamtaorta als Ldlr-/--Tiere, was eine athero-protektive Rolle von LASP1 nahelegt. Passend hierzu führte Stimulation mit oxLDL in Makrophagen zu einer Hochregulation von LASP1. Zusätzlich internalisierten LASP1-/--Makrophagen signifikant mehr oxLDL im Vergleich zu LASP1-exprimierenden Zellen. Analog zu den Daten aus der Krebsforschung konnte eine reduzierte endotheliale Adhäsion sowie chemotaktische Migration von Ldlr.LASP1-/--Monozyten im Vergleich zu Ldlr-/-- Monozyten festgestellt werden. Dies ließe isoliert betrachtet eine pro-atherogene Rolle von LASP1 vermuten. Ein Nachweis von LASP1 im Zellkern von BMDMs konnte, zusätzlich zum fehlenden Shuttelproteinpartner ZO-2, nicht erbracht werden. Die Interaktion von LASP1 mit Transkriptionsfaktoren scheint daher unwahrscheinlich. Kongruent mit diesen Ergebnissen zeigte sich keine Veränderung der Transkription, der Proteinexpression sowie Sekretion von TNF! und ADAM17 durch den LASP1-KO. Insgesamt kommt LASP1 eine zweifellos komplexe Rolle in der Atherogenese zu. Die Ergebnisse der HFD-Versuche legen nahe, dass die primär anti-atherosklerotischen Einflüsse von LASP1 in vivo gegenüber den eher pro-atherosklerotischen Effekten des Proteins in vitro überwiegen.
N2  - As of today, the regulatory scaffold protein LASP1 is mainly known from cancer research. Through its localization at dynamic actin skeletal constructs (cf. invadopodia, podosomes), LASP1 has been shown to be involved in cell migration, proliferation, and invasiveness of certain tumor cells. In 2012, LASP1 was detected in human macrophages, the protagonists of atherosclerosis. Because of a large intersection of mechanisms and signaling pathways of cancer and atherosclerosis, we further explored the role of LASP1 in the context of atherosclerosis. In a 16-week high-fat diet experiment, LASP1.Ldlr-/- mice showed more atherosclerotic lesions in the total aorta than Ldlr-/- animals, suggesting an athero-protective role of LASP1. In vitro, LASP1-/- macrophages internalized significantly more oxLDL than LASP1 positive cells. LASP1 did not shuttle in the nucleus of bone marrow derived macrophages. Thus, the interaction of LASP1 with transcription factors seems unlikely. Congruent with these results, LASP1-KO had no effects on the transcription, protein expression, or secretion of TNFα and ADAM17.

In accordance with the data from cancer research, reduced endothelial adhesion as well as chemotactic migration of Ldlr.LASP1-/- monocytes was detected compared to Ldlr-/-- monocytes. These findings on the other hand would suggest a pro-atherogenic role of LASP1.

Overall, LASP1 undoubtedly has a complex role in atherogenesis. The results of the HFD experiments suggest that the primarily anti-atherosclerotic influences of LASP1 in vivo predominate over the more pro-atherosclerotic effects of the protein in vitro.
KW  - Arteriosklerose
KW  - Schaumzelle
KW  - Maus
KW  - Monozyt
KW  - lasp
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239353
ER  - 
TY  - THES
A1  - Friedrich, Anna-Lena
T1  - FoxO3-mediated, inhibitory effects of CNP on the profibrotic activation of lung fibroblasts
T1  - FoxO3-vermittelte, hemmende Wirkungen von CNP auf die profibrotische Aktivierung von Lungenfibroblasten
N2  - Idiopathic Pulmonary Fibrosis (IPF) is a progressive parenchymal lung disease with limited therapeutic treatments. Pathologically altered lung fibroblasts, called myofibroblasts, exhibit increased proliferation, migration, and collagen production, and drive IPF development and progression. Fibrogenic factors such as Platelet derived growth factor-BB (PDGF-BB) contribute to these pathological alterations. Endogenous counter-regulating factors are barely known. Published studies have described a protective role of exogenously administered C-type Natriuretic Peptide (CNP) in pathological tissue remodeling, for example in heart and liver fibrosis. CNP and its cyclic GMP producing guanylyl cyclase B (GC-B) receptor are expressed in the lungs, but it is unknown whether CNP can attenuate lung fibrosis by this pathway. To address this question, we performed studies in primary cultured lung fibroblasts.
To examine the effects of the CNP/GC-B pathway on PDGF-BB-induced collagen 
production, proliferation, and migration in vitro, lung fibroblasts were cultured from wildtype control and GC-B knockout mice. Human lung fibroblasts from patients with IPF and healthy controls were obtained from the UGMLC Biobank. In RIA experiments, CNP, at 10nM and 100nM, markedly and similarly increased cGMP levels in both the murine and human lung fibroblasts, demonstrating GC-B/cGMP signaling. CNP reduced PDGF-BB induced proliferation and migration of lung fibroblasts in BrdU incorporation and gap closure assays, respectively. CNP strongly decreased PDGF-BB-induced collagen 1/3 expression as measured by immunocytochemistry and immunoblotting. Importantly, the protective actions of CNP were preserved in IPF fibroblasts. It is known that the profibrotic actions of PDGF-BB are partly mediated by phosphorylation and nuclear export of Forkhead Box O3 (FoxO3), a transcription factor downregulated in IPF. CNP prevented PDGF-BB elicited FoxO3 phosphorylation and nuclear exclusion in both murine and human control and IPF fibroblasts. CNP signaling and functions were abolished in GC-B-deficient lung fibroblasts. 
Taken together, the results show that CNP moderates the PDGF-BB-induced activation and differentiation of human and murine lung fibroblasts to myofibroblasts. This effect is mediated CNP-dependent by GC-B/cGMP signaling and FoxO3 regulation. To follow up the patho-physiological relevance of these results, we are generating mice with fibroblast-restricted GC-B deletion for studies in the model of bleomycin-induced pulmonary fibrosis.
N2  - Idiopathische pulmonale Fibrose (IPF) ist eine progressiv fortschreitende, parenchymale Lungenerkrankung mit beschränkten therapeutischen Behandlungsmöglichkeiten. 
Pathologisch veränderte Lungenfibroblasten, sogenannte Myofibroblasten, zeigen eine verstärkte Proliferation, Migration und Kollagenproduktion, die zu einem permanenten Fortschreiten der Erkrankung führen. Während fibrogene Faktoren wie Platelet derived growth factor-BB (PDGF-BB) zu dieser pathologischen Veränderung beitragen, sind endogene Faktoren, die diesem Wandel entgegenwirken, kaum bekannt. Allerdings konnten Studien bereits eine protektive Wirkung von exogen verabreichten C-typ natriuretischen Peptid (CNP) auf krankhaft verändertes Gewebe beschreiben, wie beispielsweise bei Herz- und Leberfibrose. Es ist bekannt, dass CNP und sein cGMP produzierender Rezeptor Guanylyl-Cyclase-B Rezeptor (GC-B) in der Lunge exprimiert werden. Allerdings konnte noch nicht nachgewiesen werden, ob CNP durch diesen Signalweg den Fortschritt einer Lungenfibrose verzögern kann. Um dies herauszufinden, wurden Experimente mit kultivierten, primären Lungenfibroblasten durchgeführt. 
Um die Effekte des CNP/ GC-B Signalwegs auf die PDGF-BB induzierte Kollagenproduktion, Proliferation und Migration in vitro zu überprüfen, wurden 
Lungenfibroblasten von Kontroll- und GC-B-Knock-Out Mäusen kultiviert. Weiterhin 
erhielten wir von der UGMLC Biobank menschliche Fibroblasten von IPF-erkrankten
Patienten, sowie gesunde Kontrollfibroblasten. Zur Bestätigung der Funktionalität des GC-B/cGMP Weges, wurde in murinen und humanen Fibroblasten mithilfe eines RIAs 
ein durch CNP (10nM und 100nM) deutlich erhöhtes cGMP-Level gemessen. CNP reduzierte die durch PDGF-BB induzierte Beschleunigung von Proliferation und 
Migration der Lungenfibroblasten, was mit Hilfe von BrdU incorporation und Gap closure assay nachgewiesen wurde. Ebenfalls zeigten Immunocytochemistrie und -blotting, dass CNP den PDGF-BB induzierten Anstieg an Kollagenexpression verhindert. Somit wurde festgestellt, dass der schützende Effekt von CNP auch in IPF Fibroblasten erhalten bleibt. Weiterhin ist bekannt, dass die PDGF-BB-induzierten, profibrotischen Veränderungen durch Phosphorylierung und Export des Transkriptionsfaktors Forkhead Box O3 (FoxO3) aus dem Zellkern vermittelt werden, welcher in IPF Fibroblasten vermindert exprimiert ist. CNP verhindert diese PDGF-BB aktivierte Phosphorylierung und Translokation in murinen und humanen Kontroll- und IPF-Fibroblasten. In Lungenfibroblasten mit Deletion des GC-B- Rezeptors war das CNP-Signal und auch dessen Effekt ausgelöscht. 
Zusammengefasst zeigen die Ergebnisse, dass CNP die PDGF-BB induzierte Aktivierung und Differenzierung von menschlichen und murinen Lungenfibroblasten zu Myofibroblasten beeinflusst. Dieser Effekt wird CNP-abhängig durch den GC-B/cGMP 
Signalweg und durch die Regulierung von FoxO3 vermittelt. Um abschließend die 
pathophysiologische Relevanz dieser Erkenntnisse zu zeigen, werden zukünftig Mäuse 
mit einer fibroblasten-spezifischen Deletion des GC-B Rezeptors für Studien in 
Bleomycin-induzierter Lungenfibrose genutzt.
KW  - Idiopathische pulmonale Fibrose
KW  - Transkriptionsfaktor
KW  - Natriuretisches Hormon
KW  - C-type natriuretic peptide
KW  - FoxO3
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359845
ER  - 
TY  - THES
A1  - Menger, Kristina Rebekka
T1  - Bauchlagerung für nicht-intubierte ARDS- und COVID-19 Patient/innen: eine systematische Übersichtsarbeit
T1  - Prone Positioning for non-intubated ARDS and COVID-19 patients: a systematic review
N2  - Die Bauchlagerung von intubierten ARDS-Patient/innen mit einer schlechten Oxygenierung wird laut Leitlinie seit mehreren Jahren als supportive Therapiemaßnahme empfohlen. Im Rahmen der COVID-19 Pandemie wurde nun erstmalig die Bauchlagerung auch bei hypoxämischen, nicht-intubierten Patient/innen untersucht. Diese Fragestellung wurde in der vorliegenden Arbeit mittels einer systematischen Übersichtsarbeit betrachtet. Aufgrund der aktuellen Pandemiesituation wurden neben ARDS-Patient/innen im Allgemeinen insbesondere COVID-19 Patient/innen mit einem akuten Lungenversagen als Subgruppe untersucht. 
Am 21.11.2020 wurde eine systematische Suche nach Studien in den Datenbanken MEDLINE, Cochrane COVID-19 Study Register und Living Overview of the Evidence  platform durchgeführt. Die Ergebnisse wurden, wo möglich, in Form einer Meta-Analyse zusammengefasst, in Tabellen darstellt oder deskriptiv beschrieben. Das Risiko für Bias wurde jeweils für die eingeschlossenen kontrollierten Studien mittels ROBINS-I beurteilt. Die Vertrauenswürdigkeit der Evidenz der gesamten Arbeit wurde mit Hilfe des GRADE-Ansatzes untersucht. 
Insgesamt wurden 30 Studien eingeschlossen, davon 4 kontrollierte Studien,  keine RCTs. In 3 der kontrollierten Studien wurde die Bauchlagerung bei COVID-19 Patient/innen untersucht, in einer bei Patient/innen mit einem anderweitig verursachten ARDS. Es ist unklar, ob die Bauchlagerung die Intubationsrate (RR = 0,92; 95% KI: 0,59 - 1,44; I² = 65%; sehr niedrige Vertrauenswürdigkeit der Evidenz), die Mortalität (RR = 0,55; 95% KI: 0,23 - 1,30; I² = 60%; sehr niedrige Vertrauenswürdigkeit der Evidenz) und die Wahrscheinlichkeit für eine Aufnahme auf die Intensivstation (RR = 0,94; 95% KI: 0,54 - 1,63; I2  = 71%; sehr niedrige Vertrauenswürdigkeit der Evidenz) verringern kann. Auch für die anderen betrachteten Endpunkte konnte kein signifikanter Effekt der Bauchlagerung nachgewiesen werden  Im Vergleich der Subgruppen „Nicht-COVID-19“ (8 Studien) und „COVID-19“ (22 Studien) konnten in Bezug auf alle betrachteten Endpunkte keine relevanten Unterschiede festgestellt werden. 
Insgesamt ist die Evidenz nicht ausreichend, um Vor- und Nachteile der Bauchlagerung für nicht-intubierte ARDS Patient/innen gegenüber der üblichen Rückenlagerung aufzuzeigen und diese für die Praxis zu empfehlen.
N2  - Prone positioning of intubated ARDS patients with poor oxygenation has been recommended as a supportive therapy for several years. In the context of the COVID-19 pandemic, prone positioning has now been investigated for the first time in hypoxemic, non-intubated patients. This issue was examined by our systematic review. Because of the current pandemic situation, we assessed the effects of prone positioning in COVID-19 patients with acute respiratory failure in particular as a subgroup in addition to ARDS patients in general. 
A systematic search for studies was performed in MEDLINE, Cochrane COVID-19 Study Register, and Living Overview of the Evidence platform databases on 21 November 2020. Where possible, results were analyzed in the form of meta-analysis, presented in tables, or described descriptively. Risk of bias was assessed for each of the included controlled studies using ROBINS-I. The quality of evidence for the entire systematic review was assessed using the GRADE approach. 
A total of 30 studies were included, 4 of which were controlled trials, but no RCTs. 3 of the controlled trials investigated prone positioning in COVID-19 patients, one trial in patients with ARDS caused by other means. It is unclear whether prone positioning can reduce intubation rate (RR =0.92; 95% CI: 0.59-1.44; I²=65%; very low quality evidence), mortality (RR =0.55; 95% CI: 0.23-1.30; I²=60%; very low quality evidence), and the likelihood of ICU admission (RR =0.94; 95% CI: 0.54-1.63; I2 =71%; very low quality evidence). No significant effect of prone positioning was demonstrated for the other outcomes considered either. When comparing the "non-COVID-19" (8 studies) and "COVID-19" (22 studies) subgroups, no relevant differences were found for all outcomes assessed. 
Overall, the evidence is insufficient to demonstrate benefits and harms of prone positioning for non-intubated ARDS patients compared with usual care and to translate the results into practice.
KW  - Bauchlage
KW  - ARDS
KW  - COVID-19
KW  - Nicht-invasive Beatmung
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359865
ER  - 
TY  - THES
A1  - Hajduk, Maurice Martin
T1  - Darf es etwas mehr sein? Neuroenhancement im Studium – eine Befragung an Würzburger Hochschulen
T1  - Can it be a little more? Neuroenhancement during studies - a survey at Würzburg universities
N2  - Neuroenhancement (NE) bezeichnet die Einnahme psychotroper Substanzen mit dem Ziel der geistigen Leistungssteigerung oder Beruhigung. NE wird durch gesunde Perso- nen genutzt. Es besteht somit keine Indikation zur Einnahme psychotroper Wirkstoffe. Zum NE genutzte Substanzen sind z.B. Koffeintabletten, verschreibungspflichtige Medi- kamente oder illegale Substanzen. Die bisherige Forschung findet Hinweise auf einen Zusammenhang zwischen NE und ADHS-Symptomen, einigen Aspekten psychischer Gesundheit, sowie Substanzkonsum. Bisher gibt es keine Forschung zu NE am Hoch- schulstandort Würzburg.
Es wurde eine anonyme online Querschnittsbefragung im ersten Quartal 2021 durchge- führt. Eingeladen waren 5600 Studierende der Julius-Maximilians-Universität Würzburg und der Hochschule für angewandte Wissenschaften Würzburg Schweinfurt. Der Frage- bogen bestand aus 53 Items und enthielt u. a. die folgenden validierten Messinstrumente: ASRS, PSS-10, PHQ-4 und AUDIT-C.
Die Response Rate lag bei 18% (n = 1011). Das Wissen über NE war weit unter den Stu- dierenden verbreitet. Die Prävalenz für Neuroenhancement im Studium lag bei 12.7%. Die drei meistgenannten Substanzen waren Koffeintabletten (6.6%), Cannabis (4.5%) und Methylphenidat (4.3%). Häufigster Anlass für NE war die Prüfungsvorbereitung. Es zeigten sich deutliche Unterschiede zwischen den Fachbereichen, u.a. hinsichtlich der Prävalenz von NE. ADHS-Symptomen, Stress, Ängstlichkeit, und Depressivität waren positiv mit NE assoziiert. Ein stärkerer Effekt ergab sich für den Zusammenhang zwi- schen NE und riskanten Alkoholkonsum bzw. Tabakkonsum. Diese Ergebnisse wurden durch eine binomial logistische Regression bestätigt.
Die konsumierten Substanzen, das Wissen über NE, die Prävalenz von NE und die Gründe für dessen Nutzung fügen sich nahtlos in die bisherige Forschung ein. Auch die Assoziation zwischen ADHS-Symptomen, Stress, Ängstlichkeit, Depressivität, riskan- tem Alkoholkonsum und Tabakkonsum bestätigt bisherige Forschungsergebnisse.
Es konnte gezeigt werden, dass rund ein Zehntel der Studierenden NE bereits genutzt haben. In Anbetracht der gesundheitlichen Gefahren, die mit NE einhergehen ist die Etab- lierung bzw. der Ausbau von Aufklärung-, Beratungs- und Hilfsangeboten für Studie- rende anzustreben sowie weitere Forschung zum Thema indiziert.
N2  - Neuroenhancement (NE) refers to the use of psychotropic substances with the aim of increasing mental performance or calming down. NE is used by healthy people. There is therefore no indication to take psychotropic substances. Substances used for NE include caffeine tablets, prescription drugs or illegal substances. Research has found evidence of a link between NE and ADHD symptoms, some aspects of mental health, and substance use. Up to now there has been no research on NE at the Würzburg university site. An anonymous online cross-sectional survey was conducted in the first quarter of 2021. 5,600 students from Julius-Maximilians-Universität Würzburg and the University of Applied Sciences Würzburg Schweinfurt were invited to participate. The questionnaire consisted of 53 items and included the following validated screening instruments: ASRS, PSS-10, PHQ-4 and AUDIT-C. The response rate was 18% (n = 1011). Knowledge about NE was widespread among the students. The prevalence of neuroenhancement during studies was 12.7%. The three most frequently mentioned substances were caffeine tablets (6.6%), cannabis (4.5%) and methylphenidate (4.3%). The most common reason for NE was exam preparation. There were clear differences between the subject areas, especially with regard to the prevalence of NE. ADHD symptoms, stress, anxiety and depression were positively associated with NE. A stronger effect was found for the relationship between NE and risky alcohol consumption or tobacco use. These results were confirmed by binomial logistic regression. The substances consumed, the knowledge about NE, the prevalence of NE and the reasons for its use fit in seamlessly with previous research. The association between ADHD symptoms, stress, anxiety, depression, risky alcohol consumption and tobacco consumption also confirms previous research findings. It was shown that around a tenth of students have already used NE. In view of the health risks associated with NE, the establishment or expansion of educational programs, counseling and support services for students is desirable and further research on the topic is indicated.
KW  - Psychische Gesundheit
KW  - Neuroenhancement
KW  - Cognitive Enhancement
KW  - Brain doping
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359812
ER  - 
TY  - THES
A1  - Hammel, Clara
T1  - Einfluss longitudinaler Veränderungen der linksventrikulären Ejektionsfraktion auf das Langzeitüberleben bei Herzinsuffizienzpatienten mit leicht reduzierter Ejektionsfraktion oder reduzierter Ejektionsfraktion
T1  - Impact of longitudinal changes in left ventricular ejection fraction on outcomes of patients with mid-range ejection fraction and reduced ejection
N2  - Diese retrospektive Studie an der Universitätsklinik Würzburg diente der Beurteilung der longitudinalen Funktion in Bezug auf die Gesamtmortalität bei Patienten mit HFmrEF und HFrEF. Die Gruppierung erfolgte anhand der jeweiligen Baseline LVEF. Eine weitere Unterteilung erfolgte in eine ischämische oder nicht-ischämische Genese der HF. Die Subgruppen wurden anhand der Baseline klinischen Charakteristika sowie der echokardiographischen Parameter verglichen. Hier ließ sich ein relativ ähnliches Patientenklientel mit vergleichbarem Alter, Geschlecht, BMI sowie kardialen Risikofaktoren zeigen. Signifikante Unterschiede ergab der Vergleich des NYHA-Stadiums, der Nierenfunktion sowie des Auftretens von Myokardinfarkten. 
Die Veränderung der LVEF über die Zeit hat einen zentralen Stellenwert zur Evaluation des Outcomes von Patienten mit HFmrEF und HFrEF. Eine Verbesserung der LVEF fand sich signifikant häufiger bei HFrEF Patienten als bei HFmrEF Patienten, welche über die Zeit signifikant häufiger eine stabile LVEF aufwiesen. 
Außerdem war nach Auswertung der Überlebenskurven nach Kaplan-Meier in HFmrEF Patienten eine verbesserte oder unveränderte LVEF über die Zeit mit einem besseren Überleben verbunden, vor allem bei Patienten mit ischämischer Ätiologie. In der HFrEF Gruppe konnte gezeigt werden, dass sowohl Patienten mit ischämischer als auch mit nicht-ischämischer Ätiologie bei Vorliegen einer verbesserten oder unveränderten LVEF über die Zeit ein besseres Outcome aufwiesen. 
Eine erniedrigte MAPSE bedeutete vor allem bei HFmrEF Patienten mit nicht-ischämischer Ätiologie ein schlechteres Outcome.
Die Ergebnisse dienten unter anderem der weiteren Charakterisierung der HFmrEF und HFrEF Gruppe sowie der Identifikation von Faktoren zur Beurteilung der Veränderung der LVEF über die Zeit und der Prognose des Langzeitüberlebens beider Gruppen. Ziel für die Zukunft sollte sein, auch für HFmrEF Patienten evidenzbasierte Herzinsuffizienz Therapien zu etablieren.
N2  - This retrospective study at the University Hospital of Wuerzburg was designed to assess longitudinal function in relation to all-cause mortality in patients with HFmrEF and HFrEF. The grouping was based on the respective baseline LVEF. A further subdivision was made using coronary angiographic data into ischemic or non-ischemic genesis of HF. The subgroups were compared on the basis of baseline clinical characteristics and echocardiographic parameters. This revealed a relatively similar patient cohort with comparable age, gender, BMI and cardiac risk factors. Significant differences were found in the comparison of NYHA stage, renal function and the occurrence of myocardial infarction. 
The change in LVEF over time is of central importance for evaluating the outcome of patients with HFmrEF and HFrEF. An improvement in LVEF was significantly more common in HFrEF patients than in HFmrEF patients, who were significantly more likely to have a stable LVEF over time.
Furthermore, according to the Kaplan-Meier survival curves in HFmrEF patients, improved or unchanged LVEF over time was associated with better survival, especially in patients with ischemic etiology. In the HFrEF group, it was shown that both patients with ischemic and non-ischemic etiology had a better outcome with improved or unchanged LVEF over time.
A lower MAPSE was associated with a worse outcome, especially in HFmrEF patients with non-ischemic etiology.
The results were used, among other things, to further characterize the HFmrEF and HFrEF groups and to identify factors for evaluating the change in LVEF over time and the prognosis of long-term survival in both groups. The aim for the future should be to establish evidence-based heart failure therapies for HFmrEF patients as well.
KW  - Transthorakale Echokardiographie
KW  - Herzinsuffizienz
KW  - MAPSE
KW  - HFmrEF
KW  - HFrEF
KW  - Langzeitüberleben
KW  - leicht reduzierte Herzinsuffizienz
KW  - reduzierte Herzinsuffizienz
KW  - longitudinale Funktion
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360025
ER  - 
TY  - JOUR
A1  - Allert, Stefanie
A1  - Förster, Toni M.
A1  - Svensson, Carl-Magnus
A1  - Richardson, Jonathan P.
A1  - Pawlik, Tony
A1  - Hebecker, Betty
A1  - Rudolphi, Sven
A1  - Juraschitz, Marc
A1  - Schaller, Martin
A1  - Blagojevic, Mariana
A1  - Morschhäuser, Joachim
A1  - Figge, Marc Thilo
A1  - Jacobsen, Ilse D.
A1  - Naglik, Julian R.
A1  - Kasper, Lydia
A1  - Mogavero, Selene
A1  - Hube, Bernhard
T1  - \(Candida\) \(albicans\)-Induced Epithelial Damage Mediates Translocation through Intestinal Barriers
JF  - mBio
N2  - Life-threatening systemic infections often occur due to the translocation of pathogens across the gut barrier and into the bloodstream. While the microbial and host mechanisms permitting bacterial gut translocation are well characterized, these mechanisms are still unclear for fungal pathogens such as Candida albicans, a leading cause of nosocomial fungal bloodstream infections. In this study, we dissected the cellular mechanisms of translocation of C. albicans across intestinal epithelia in vitro and identified fungal genes associated with this process. We show that fungal translocation is a dynamic process initiated by invasion and followed by cellular damage and loss of epithelial integrity. A screen of >2,000 C. albicans deletion mutants identified genes required for cellular damage of and translocation across enterocytes. Correlation analysis suggests that hypha formation, barrier damage above a minimum threshold level, and a decreased epithelial integrity are required for efficient fungal translocation. Translocation occurs predominantly via a transcellular route, which is associated with fungus-induced necrotic epithelial damage, but not apoptotic cell death. The cytolytic peptide toxin of C. albicans, candidalysin, was found to be essential for damage of enterocytes and was a key factor in subsequent fungal translocation, suggesting that transcellular translocation of C. albicans through intestinal layers is mediated by candidalysin. However, fungal invasion and low-level translocation can also occur via non-transcellular routes in a candidalysin-independent manner. This is the first study showing translocation of a human-pathogenic fungus across the intestinal barrier being mediated by a peptide toxin. IMPORTANCE Candida albicans, usually a harmless fungus colonizing human mucosae, can cause lethal bloodstream infections when it manages to translocate across the intestinal epithelium. This can result from antibiotic treatment, immune dysfunction, or intestinal damage (e.g., during surgery). However, fungal processes may also contribute. In this study, we investigated the translocation process of C. albicans using in vitro cell culture models. Translocation occurs as a stepwise process starting with invasion, followed by epithelial damage and loss of epithelial integrity. The ability to secrete candidalysin, a peptide toxin deriving from the hyphal protein Ece1, is key: C. albicans hyphae, secreting candidalysin, take advantage of a necrotic weakened epithelium to translocate through the intestinal layer.
KW  - Candida albicans
KW  - candidalysin
KW  - host cell damage
KW  - host cell invasion
KW  - intestinal barrier
KW  - necrosis
KW  - translocation
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221084
VL  - 9
IS  - 3
ER  -