TY - JOUR A1 - Tesfamariam, Y. A1 - Jakob, T. A1 - Wöckel, A. A1 - Adams, A. A1 - Weigl, A. A1 - Monsef, I. A1 - Kuhr, K. A1 - Skoetz, N. T1 - Adjuvant bisphosphonates or RANK-ligand inhibitors for patients with breast cancer and bone metastases: A systematic review and network meta-analysis JF - Critical Reviews in Oncology / Hematology N2 - Bone-modifying agents like bisphosphonates and receptor activator of nuclear factor kappaβ ligand (RANK-L) inhibitors are used as supportive treatments in breast cancer patients with bone metastases to prevent skeletal-related events (SREs). Due to missing head-to-head comparisons, a network meta-analysis was performed to provide a hierarchy of these therapeutic options. Through a systematic literature search, 21 randomized controlled trials (RCTs) that fulfilled the inclusion criteria were identified. To prevent SREs, the ranking through P-scores showed denosumab (RR: 0.62; 95%CI: 0.50-0.76), zoledronic acid (RR: 0.72; 95%CI: 0.61-0.84) and pamidronate (RR: 0.76; 95%CI: 0.67-0.85) to be significantly superior to placebo. Due to insufficient or heterogeneous data, overall survival, quality of life, pain response and adverse events were not able to be analyzed within the network. Although data were sparse on adverse events, the risk of significant adverse events appeared low. The results of this review can therefore be used to formulate clinical studies more precisely in order to standardise and focus on patient-relevant outcomes. KW - bisphosphonates KW - RANK-L inhibitors KW - SREs KW - RCTs Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-240827 VL - 137 ER - TY - JOUR A1 - Brinker, Titus J. A1 - Hekler, Achim A1 - Hauschild, Axel A1 - Berking, Carola A1 - Schilling, Bastian A1 - Enk, Alexander H. A1 - Haferkamp, Sebastian A1 - Karoglan, Ante A1 - von Kalle, Christof A1 - Weichenthal, Michael A1 - Sattler, Elke A1 - Schadendorf, Dirk A1 - Gaiser, Maria R. A1 - Klode, Joachim A1 - Utikal, Jochen S. T1 - Comparing artificial intelligence algorithms to 157 German dermatologists: the melanoma classification benchmark JF - European Journal of Cancer N2 - Background Several recent publications have demonstrated the use of convolutional neural networks to classify images of melanoma at par with board-certified dermatologists. However, the non-availability of a public human benchmark restricts the comparability of the performance of these algorithms and thereby the technical progress in this field. Methods An electronic questionnaire was sent to dermatologists at 12 German university hospitals. Each questionnaire comprised 100 dermoscopic and 100 clinical images (80 nevi images and 20 biopsy-verified melanoma images, each), all open-source. The questionnaire recorded factors such as the years of experience in dermatology, performed skin checks, age, sex and the rank within the university hospital or the status as resident physician. For each image, the dermatologists were asked to provide a management decision (treat/biopsy lesion or reassure the patient). Main outcome measures were sensitivity, specificity and the receiver operating characteristics (ROC). Results Total 157 dermatologists assessed all 100 dermoscopic images with an overall sensitivity of 74.1%, specificity of 60.0% and an ROC of 0.67 (range = 0.538–0.769); 145 dermatologists assessed all 100 clinical images with an overall sensitivity of 89.4%, specificity of 64.4% and an ROC of 0.769 (range = 0.613–0.9). Results between test-sets were significantly different (P < 0.05) confirming the need for a standardised benchmark. Conclusions We present the first public melanoma classification benchmark for both non-dermoscopic and dermoscopic images for comparing artificial intelligence algorithms with diagnostic performance of 145 or 157 dermatologists. Melanoma Classification Benchmark should be considered as a reference standard for white-skinned Western populations in the field of binary algorithmic melanoma classification. KW - benchmark KW - artificial intelligence KW - deep learning KW - melanoma Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220569 VL - 111 ER - TY - JOUR A1 - Brinker, Titus J. A1 - Hekler, Achim A1 - Enk, Alexander H. A1 - Berking, Carola A1 - Haferkamp, Sebastian A1 - Hauschild, Axel A1 - Weichenthal, Michael A1 - Klode, Joachim A1 - Schadendorf, Dirk A1 - Holland-Letz, Tim A1 - von Kalle, Christof A1 - Fröhling, Stefan A1 - Schilling, Bastian A1 - Utikal, Jochen S. T1 - Deep neural networks are superior to dermatologists in melanoma image classification JF - European Journal of Cancer N2 - Background Melanoma is the most dangerous type of skin cancer but is curable if detected early. Recent publications demonstrated that artificial intelligence is capable in classifying images of benign nevi and melanoma with dermatologist-level precision. However, a statistically significant improvement compared with dermatologist classification has not been reported to date. Methods For this comparative study, 4204 biopsy-proven images of melanoma and nevi (1:1) were used for the training of a convolutional neural network (CNN). New techniques of deep learning were integrated. For the experiment, an additional 804 biopsy-proven dermoscopic images of melanoma and nevi (1:1) were randomly presented to dermatologists of nine German university hospitals, who evaluated the quality of each image and stated their recommended treatment (19,296 recommendations in total). Three McNemar's tests comparing the results of the CNN's test runs in terms of sensitivity, specificity and overall correctness were predefined as the main outcomes. Findings The respective sensitivity and specificity of lesion classification by the dermatologists were 67.2% (95% confidence interval [CI]: 62.6%–71.7%) and 62.2% (95% CI: 57.6%–66.9%). In comparison, the trained CNN achieved a higher sensitivity of 82.3% (95% CI: 78.3%–85.7%) and a higher specificity of 77.9% (95% CI: 73.8%–81.8%). The three McNemar's tests in 2 × 2 tables all reached a significance level of p < 0.001. This significance level was sustained for both subgroups. Interpretation For the first time, automated dermoscopic melanoma image classification was shown to be significantly superior to both junior and board-certified dermatologists (p < 0.001). KW - deep learning KW - melanoma KW - skin cancer KW - artificial intelligence Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220539 VL - 119 ER - TY - THES A1 - Zuber, Jonas Maximilian T1 - Evaluation von Sedierungen und Allgemeinanästhesien zur Durchführung bildgebender Verfahren bei Säuglingen bis zum 6. Lebensmonat T1 - Evaluation of sedation and general anesthesia for performing imaging procedures in infants up to 6 months of age N2 - Vorliegende Untersuchung am Universitätsklinikum Würzburg sowie die Befragung von Anästhesisten/Anästhesistinnen im Raum der 3 DACH-Länder zeigen, dass bildgebende Verfahren bei Säuglingen mit einer niedrigen Rate an Komplikationen, zumeist in medikamentöser Sedierung mit Propofol, durchgeführt werden. Wie international üblich ist im Säuglingsalter die Magnetresonanztomographie das bildgebende Verfahren der Wahl und wird, mit überzeugender Häufigkeit, erfolgreich durchgeführt. Die Untersuchung am Universitätsklinikum Würzburg legt nahe, dass männliche Säuglinge häufiger eine Bildgebung benötigen und häufiger höheren ASA-Kategorie zugeschrieben werden. Dabei scheinen sie auch häufiger Komplikationen zu erleben und bedürfen daher besonderer Aufmerksamkeit. Eine eventuelle Alternative zur Sedierung kann dabei die „feed-and-sleep“ Methode darstellen. In unserer Umfrage konnten wir erheben, dass diese Methode bisher wenig verbreitet ist, obwohl in diesem Zusammenhang eventuell Abläufe und Prozesszeiten strukturiert und optimiert werden können, da beispielsweise die Nachüberwachung entfällt. Vorstellbar wäre beispielsweise, mehrere Säuglinge zum gleichen Zeitpunkt ins MRT zu bestellen, um gegebenenfalls den am frühesten eingeschlafenen Säugling vorzuziehen. Diese Methode sollte zukünftig Einzug in die wissenschaftliche Untersuchung von bildgebenden Verfahren bei Säuglingen finden. Die Umfrage im deutschsprachigen Raum zeigt eine Leitlinien-gerechte Betreuung von Säuglingen für bildgebende Verfahren, die mit einer hohen Qualität, und zumeist erfolgreich von erfahrenen Anästhesisten/Anästhesistinnen durchgeführt wird. Eventuelle Verbesserungen können im Bereich der Ausbildung nachfolgender Ärztinnen/Ärzte und in der häufigeren Verwendung der „feed-and-sleep“ Methode liegen, die vielen Kollegen/Kolleginnen bekannt ist, aber nur selten durchgeführt wird. Ziel ist eine qualitativ hochwertige, schnellstmöglich durchgeführte Bildgebung, die ohne oder mit der niedrigst möglichen Dosierung eines sedierenden Medikamentes zu erreichen ist. N2 - The present study at the University Hospital of Würzburg and the survey of anesthesiologists in the three DACH countries show that imaging procedures are carried out in infants with a low rate of complications, mostly under medical sedation with propofol. As is common practice internationally, magnetic resonance imaging is the imaging method of choice in infancy and is mostly carried out successfully. The study at the University Hospital of Würzburg suggests that male infants require imaging procedures more frequently and are assigned to higher ASA categories in comparison with female infants.They also seem to experience complications more often and therefore require special attention. The “feed and sleep” method can be a potential alternative to sedation. In our survey, we found that this method is not yet used widely, although in this context processes and process times can potentially be restructured and optimized since, for example, follow-up monitoring is no longer necessary. It would be conceivable, for example, to order several infants for an MRI at the same time in order to prioritize the infant who fell asleep earliest. In the future, this method should find its way into the scientific study of imaging procedures in infants. The survey in German-speaking countries shows guideline-compliant care of infants for imaging procedures, which is carried out with high quality and mostly successfully by experienced anesthesiologists. Possible improvements could lie in the education of doctors in training and in the more frequent use of the “feed-and-sleep” method, which is known to many colleagues but is rarely carried out. The goal is a high-quality imaging that is performed as quickly as possible and can be achieved without or with the lowest possible dosage of a sedative medication. KW - Sedierung KW - Säugling KW - Narkose KW - Kernspintomografie KW - feed-and-sleep KW - Umfrage KW - Anästhesie KW - Monitoring Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-361111 ER - TY - THES A1 - Dalkmann, Theresa T1 - Evaluierung prognostischer und prädiktiver Biomarker beim neoadjuvant vorbehandelten Rektumkarzinom T1 - Evaluation of prognostic and predictive biomarkers for neoadjuvant chemoradiotherapy in locally advanced rectal cancer (LARC) N2 - Fragestellung. Osteopontin (OPN) kann im Blut nachgewiesen werden und wird bei vielen Tumorentitäten exprimiert, wie auch der Tyrosinkinaserezeptor c-Met und sein Ligand, das Zytokin Hepatocyte Growth Factor (HGF). In der vorliegenden Arbeit untersuchten wir die prognostische und prädiktive Wertigkeit der Plasmakonzentrationen von OPN, c-Met und HGF bei Patienten mit lokal fortgeschrittenem Rektumkarzinom (LARC). Methodik. Das Plasma von 63 Patienten mit LARC wurde untersucht. Die Blutentnahmen (EDTA-Plasma) erfolgten vor Therapiebeginn sowie im Verlauf. Die Plasmaspiegel von OPN, c-Met und HGF wurden mittels Enzyme-Linked Immunosorbent Assay analysiert. Die Konzentrationen wurden auf eine Korrelation mit den klinischen Parametern untersucht. Ergebnisse. 68 Patienten wurden neoadjuvant mit einer Radiochemotherapie behandelt, 63 Blutproben wurden untersucht. Initial befanden sich nach UICC 14 Patienten in Stadium II, 47 in Stadium III und 7 in Stadium IV. Das mediane Follow-Up betrug 29,87 Monate. 20 der 68 Patienten (29,4 %) verstarben, 19 entwickelten Fernmetastasen. OPN korrelierte signifikant mit dem Überleben (p=0,001). OPN-Werte korrelierten mit dem pT-Stadium (R:0,445 p=0,018) und dem pUICC-Stadium (R:0,412 p=0,018), sowie mit dem Auftreten von Fernmetastasen (R:0,271 p=0,031). Eine Korrelation zwischen OPN und dem Therapieansprechen konnte gezeigt werden: pathologisch komplette Remission (pCR) (R:0,379 p=0,001), NAR-Score (R:0,373 p=0,015), TRG (R:0,380 p=0,020). Die logistische Regressionsanalyse ergab eine Prädiktivität OPNs für pCR (OR:0,990 p=0,009), NAR-Score (OR:1,008 p=0,007), TRG (OR:0,459 p=0,008). C-Met und HGF korrelierten nicht mit dem Überleben. Für c-Met und HGF ergab sich keine Korrelation zu initialen klinischen Daten und Therapieansprechen. Die logistische Regression ergab keinen prädiktiven Wert. Schlussfolgerung. Die Plasmakonzentration von OPN besitzt prognostische und prädiktive Wertigkeit beim LARC. Die Konzentrationen von c-Met und HGF sind nicht prognostisch für das Überleben oder prädiktiv für das Therapieansprechen. N2 - Purpose. The glycoprotein Osteopontin (OPN), tyrosine kinase receptor c-Met and it´s ligand Hepatocyte Growth Factor (HGF) can be detectet in blood and are known to be overexpressed in many kinds of human cancer. Here we examine their prognostic and predictive value in patients with locally advanced rectal cancer (LARC). Patients and methods. In a monocentric prospective study EDTA-plasma was drawn from patients who received neoadjuvant chemoradiotherapy (CRT) in LARC. Blood samples were taken before CRT and at different timepoints during the follow-up. We used an Enzyme-linked Immunosorbent Assay (ELISA) to analyse the plasma concentrations of OPN, c-Met and HGF. Results. 68 patients (48 males, 20 females) were included. Blood samples were drawn from 63 patients. Initially, 14 patients had UICC stage II, 47 had UICC stage III and 7 had UICC stage IV. Median follow-up was 29,9 months. 20 out of 68 patients died during follow-up (29,4 %), 19 developed metastasis (27,9 %). Patients with OPN median (91,8 % vs. 58,6 %, p = 0,001). Higher OPN-concentrations were correlated with pT (R = 0,445, p = 0,018), postoperative UICC (R = 0,412, p = 0,018) and metastasis (R = 0,271, p = 0,031). Pretherapeutical OPN levels were significantly different depending on the response to CRT: 415,9 ± 62,6 ng/ml in patients with pathological complete response (pCR) vs. 703,3 ± 285,9 ng/ml in patients with pathological incomplete response (pIR), p < 0,001). Logistic regression showed a predictive value of OPN for pCR (OR = 0,990, p = 0,009). Pretherapeutic c-Met and HGF concentrations were not associated with survival rates. There was no correlation between initial c-Met or HGF and clinical characteristics. There was von predictive value for c-Met or HGF. Conclusion. Osteopontin plasma levels might have prognostic and predictive value in LARC. We could not find a prognostic or predictive significance for c-Met or HGF. KW - Biomarker KW - Neoadjuvant vorbehandeltes Rektumkarzinom KW - prognostische und prädiktive Biomarker KW - prognostic and predictive biomarker KW - locally advanced rectal cancer KW - neoadjuvant radiochemotherapy KW - Mastdarmkrebs KW - Rektumkarzinom KW - Neodadjuvant KW - prognostische KW - prädiktive Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-363368 ER - TY - THES A1 - Junghanns, Lara Madeleine T1 - Resistenzmechanismen gegen Amphotericin B in humanpathogenen Hefepilzen T1 - Resistance mechanism to amphotericin B in human pathogenic yeasts N2 - Die 2009 erstmals entdeckte Spezies C. auris erlangte binnen kürzester Zeit zunehmend weltweite Aufmerksamkeit. Vor allem die Tendenz der Multiresistenzentwicklung und das rasche Auslösen von nosokomialen Infektionen erschweren den Umgang und die Therapie von C. auris Infektionen im Vergleich zu anderen Candida Spezien. Diese Dissertationsarbeit umfasst eine systematische Resistenzanalyse der im NRZMyk vorhandenen Stammsammlung aus C. auris und C. parapsilosis Isolaten, um Aufschluss über den Wirkmechanismus von Amphotericin B in Hefepilzen zu erlangen. Anhand der zunächst durchgeführten Amphotericin B-Resistenztestungen kristallisierten sich CAU37 und CAU43 mit MHK-Werten bis zu 12 µg/ml als stark Amphotericin B-resistente Isolate heraus. Die Analyse der Sequenzierungsergebnisse zeigte bei beiden Stämmen eine Mutation im ERG4 Gen an Position 576, welche nicht eindeutig als alleinige Ursache für die verminderte Amphotericin B-Empfindlichkeit festgelegt werden konnte. Dennoch wurde im Rahmen eines Survival Assays bei beiden Amphotericin B-resistenten Isolaten anfänglich eine konzentrationsabhängige Aktivität gegenüber Amphotericin B festgestellt, bevor ein Nachwachsen der Kulturen beobachtet wurde. Somit wurde die Vermutung aufgestellt, dass lediglich ein Teil der aufgebrachten Candida-Zellen abgetötet wird und dies in einer Vermehrung der überlebenden Zellen resultiert. Des Weiteren konnte im Rahmen von Resistenztestungen mit dem Sphingolipidinhibitor Myriocin nachgewiesen werden, dass vor allem in Amphotericin B-resistenten Isolaten eine deutliche Wirkungsverstärkung des Polyens hervorgerufen wird. Diese Sensitivitätssteigerung ist allgemein bei allen C. auris Isolaten zu beobachten, fällt bei resistenten Stämmen jedoch deutlich stärker aus. Hierdurch kam die Annahme auf, dass Amphotericin B-Resistenzen auch in möglichen Veränderungen des Sphingolipid-Haushaltes begründet sein könnten. Darüber hinaus scheint Myriocin keinen Einfluss auf Fluconazol-resistente oder FKS-mutierte Echinocandin-resistente C. auris Stämme zu haben. Das ebenfalls untersuchte und von Myriocin abgeleitete Medikament Fingolimod hatte jedoch ebenfalls keinen wirkungsverstärkenden Effekt. Allerdings reagierte ein Großteil der C. auris Isolate (57,6 %) sensitiv gegenüber dem neusten medizinisch bekannten Triazol Isavuconazol und es konnte erstmalig ein ECV-Wert von 0,03125 µg/ml festgelegt werden. Ein valider Vergleich von C. auris zu C. parapsilosis war aufgrund der mangelnden Anzahl an C. parapsilosis Isolaten jedoch nicht möglich N2 - The species C. auris, which was first discovered in 2009, quickly attracted worldwide attention. In particular, the development of multidrug resistance and the rapid onset of nosocomial infections complicate the management and treatment of C. auris infections compared to other Candida species. This dissertation comprises a systematic resistance analysis of the strain collection available at the NRZMyk from C. auris and C. parapsilosis isolates in order to shed light on the mechanism of action of amphotericin B in yeast fungi. CAU37 and CAU43 ermerged as highly amphotericin B-resistant isolates in the initially performed amphotericin B resistance tests, with MIC values up to 12 µg/ml. Sequencing results showed a mutation in the ERG4 gene at position 576 in both strains, which can`t be clearly identified as the main cause of the reduced susceptibility to amphotericin B. Nevertheless both amphotericin B-resistant isolates initially showed a concentration dependent activity against amphotericin B, followed by a regrowth of the cultures. The hypothesis is, that only some of the applied Candida cells are killed, resulting in a proliferation of the surviving cells. Furthermore the resistance tests with the sphingolipid inhibitor Myriocin in combination with amphotericin B showed that sublethal myriocin concentrations increased the C. auris susceptibility to amphotericin B. This increase in sensitivity is generally observed in all C. auris isolates, but is significantly stronger in resistant strains. This leeds to the assumption that amphotericin B resistance can also be due to possible changes in the sphingolipid balance. Furthermore, myriocin does not appear to have any influence on fluconazole-resistant or FKS-mutated echinocandin-resistant C. auris strains. Fingolimod, a drug also investigated and derived from Myriocin, doesn`t have any enhancing effect either. However the majority of C. auris isolates (57.6 %) reacted sensitively to the latest medically known triazole isavuconazole and for the first time an ECV value of of 0.03125 µg/ml could be determined. A valid comparison of C. auris to C. parapsilosis was not possible due to the lack of C. parapsilosis isolates. KW - Candida KW - antifungal susceptibility KW - Multidrug-Resistenz KW - Amphotericin B KW - Sphingolipide KW - Fingolimod KW - Candida auris KW - Multiresistenz KW - Myriocin KW - Isavuconazol KW - Antimykotikaresistenz KW - C. auris KW - Empdindlichkeitsprüfung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369861 ER - TY - JOUR A1 - Jeanclos, Elisabeth A1 - Albersen, Monique A1 - Ramos, Rúben J. J. A1 - Raab, Annette A1 - Wilhelm, Christian A1 - Hommers, Leif A1 - Lesch, Klaus-Peter A1 - Verhoeven-Duif, Nanda M. A1 - Gohla, Antje T1 - Improved cognition, mild anxiety-like behavior and decreased motor performance in pyridoxal phosphatase-deficient mice JF - BBA - Molecular Basis of Disease N2 - Pyridoxal 5′-phosphate (PLP) is an essential cofactor in the catalysis of ~140 different enzymatic reactions. A pharmacological elevation of cellular PLP concentrations is of interest in neuropsychiatric diseases, but whole-body consequences of higher intracellular PLP levels are unknown. To address this question, we have generated mice allowing a conditional ablation of the PLP phosphatase PDXP. Ubiquitous PDXP deletion increased PLP levels in brain, skeletal muscle and red blood cells up to 3-fold compared to control mice, demonstrating that PDXP acts as a major regulator of cellular PLP concentrations in vivo. Neurotransmitter analysis revealed that the concentrations of dopamine, serotonin, epinephrine and glutamate were unchanged in the brains of PDXP knockout mice. However, the levels of γ-aminobutyric acid (GABA) increased by ~20%, demonstrating that elevated PLP levels can drive additional GABA production. Behavioral phenotyping of PDXP knockout mice revealed improved spatial learning and memory, and a mild anxiety-like behavior. Consistent with elevated GABA levels in the brain, PDXP loss in neural cells decreased performance in motor tests, whereas PDXP-deficiency in skeletal muscle increased grip strength. Our findings suggest that PDXP is involved in the fine-tuning of GABA biosynthesis. Pharmacological inhibition of PDXP might correct the excitatory/inhibitory imbalance in some neuropsychiatric diseases. KW - pyridoxal phosphatase KW - vitamin B6 KW - γ-Aminobutyric acid (GABA) KW - motor performance KW - neuropsychiatric diseases KW - neurotransmitter biosynthesis Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323396 VL - 1865 ER - TY - JOUR A1 - Müller, Laura S. M. A1 - Cosentino, Raúl O. A1 - Förstner, Konrad U. A1 - Guizetti, Julien A1 - Wedel, Carolin A1 - Kaplan, Noam A1 - Janzen, Christian J. A1 - Arampatzi, Panagiota A1 - Vogel, Jörg A1 - Steinbiss, Sascha A1 - Otto, Thomas D. A1 - Saliba, Antoine-Emmanuel A1 - Sebra, Robert P. A1 - Siegel, T. Nicolai T1 - Genome organization and DNA accessibility control antigenic variation in trypanosomes JF - Nature N2 - Many evolutionarily distant pathogenic organisms have evolved similar survival strategies to evade the immune responses of their hosts. These include antigenic variation, through which an infecting organism prevents clearance by periodically altering the identity of proteins that are visible to the immune system of the host1. Antigenic variation requires large reservoirs of immunologically diverse antigen genes, which are often generated through homologous recombination, as well as mechanisms to ensure the expression of one or very few antigens at any given time. Both homologous recombination and gene expression are affected by three-dimensional genome architecture and local DNA accessibility2,3. Factors that link three-dimensional genome architecture, local chromatin conformation and antigenic variation have, to our knowledge, not yet been identified in any organism. One of the major obstacles to studying the role of genome architecture in antigenic variation has been the highly repetitive nature and heterozygosity of antigen-gene arrays, which has precluded complete genome assembly in many pathogens. Here we report the de novo haplotype-specific assembly and scaffolding of the long antigen-gene arrays of the model protozoan parasite Trypanosoma brucei, using long-read sequencing technology and conserved features of chromosome folding4. Genome-wide chromosome conformation capture (Hi-C) reveals a distinct partitioning of the genome, with antigen-encoding subtelomeric regions that are folded into distinct, highly compact compartments. In addition, we performed a range of analyses—Hi-C, fluorescence in situ hybridization, assays for transposase-accessible chromatin using sequencing and single-cell RNA sequencing—that showed that deletion of the histone variants H3.V and H4.V increases antigen-gene clustering, DNA accessibility across sites of antigen expression and switching of the expressed antigen isoform, via homologous recombination. Our analyses identify histone variants as a molecular link between global genome architecture, local chromatin conformation and antigenic variation. KW - histone variants KW - genome architecture KW - single molecule real time (SMRT) KW - brucei genome KW - distance-dependent decay Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224265 VL - 563 ER - TY - JOUR A1 - Munz, Matthias A1 - Richter, Gesa M. A1 - Loos, Bruno G. A1 - Jepsen, Søren A1 - Divaris, Kimon A1 - Offenbacher, Steven A1 - Teumer, Alexander A1 - Holtfreter, Birte A1 - Kocher, Thomas A1 - Bruckmann, Corinna A1 - Jockel-Schneider, Yvonne A1 - Graetz, Christian A1 - Munoz, Loreto A1 - Bhandari, Anita A1 - Tennstedt, Stephanie A1 - Staufenbiel, Ingmar A1 - van der Velde, Nathalie A1 - Uitterlinden, André G. A1 - de Groot, Lisette C. P. G. M. A1 - Wellmann, Jürgen A1 - Berger, Klaus A1 - Krone, Bastian A1 - Hoffmann, Per A1 - Laudes, Matthias A1 - Lieb, Wolfgang A1 - Andre, Franke A1 - Dommisch, Henrik A1 - Erdmann, Jeanette A1 - Schaefer, Arne S. T1 - Genome-wide association meta-analysis of coronary artery disease and periodontitis reveals a novel shared risk locus JF - Scientific Reports N2 - Evidence for a shared genetic basis of association between coronary artery disease (CAD) and periodontitis (PD) exists. To explore the joint genetic basis, we performed a GWAS meta-analysis. In the discovery stage, we used a German aggressive periodontitis sample (AgP-Ger; 680 cases vs 3,973 controls) and the CARDIoGRAMplusC4D CAD meta-analysis dataset (60,801 cases vs 123,504 controls). Two SNPs at the known CAD risk loci ADAMTS7 (rs11634042) and VAMP8 (rs1561198) passed the pre-assigned selection criteria (PAgP-Ger < 0.05; PCAD < 5 × 10−8; concordant effect direction) and were replicated in an independent GWAS meta-analysis dataset of PD (4,415 cases vs 5,935 controls). SNP rs1561198 showed significant association (PD[Replication]: P = 0.008 OR = 1.09, 95% CI = [1.02–1.16]; PD [Discovery + Replication]: P = 0.0002, OR = 1.11, 95% CI = [1.05–1.17]). For the associated haplotype block, allele specific cis-effects on VAMP8 expression were reported. Our data adds to the shared genetic basis of CAD and PD and indicate that the observed association of the two disease conditions cannot be solely explained by shared environmental risk factors. We conclude that the molecular pathway shared by CAD and PD involves VAMP8 function, which has a role in membrane vesicular trafficking, and is manipulated by pathogens to corrupt host immune defense. KW - vesicle-associated membrane protein 8 (VAMP8) KW - ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS7) KW - shared genetic basis KW - genome-wide association studies (GWAS) KW - GWAS meta-analysis Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231647 VL - 8 ER - TY - THES A1 - Pätzel [geb. Ditter], Katharina Sabine T1 - Molekulare Charakterisierung eines Mitgliedes der TNF-Rezeptor-Superfamilie des Fuchsbandwurmes \(Echinococcus\) \(multilocularis\) T1 - Molecular characterization of a TNF-receptor-superfamily member of \(Echinococcus\) \(multilocularis\) N2 - Die alveoläre Echinokokkose (AE), die durch den Fuchsbandwurm Echinococcus multilocularis verursacht wird, ist eine seltene jedoch schwere und oft tödlich verlaufende Erkrankung. Aufgrund der späten Diagnosestellung sind kurative Behandlungsmethoden häufig nicht durchführbar und als einzige Behandlungsmöglichkeit bleibt eine lebenslange und nebenwirkungsreiche Therapie mit Benzimidazolen. Verbesserte Therapieoptionen durch die Entwicklung neuer Medikamente sind dringend notwendig. Hierfür kann es hilfreich sein die Biologie des Fuchsbandwurmes und die Kommunikationswege zwischen Parasit und Wirt zu verstehen. Bereits in vorherigen Arbeiten als auch in dieser Arbeit erwiesen sich evolutionsgeschichtlich konservierte Signalwege als Kommunikationsweg zwischen dem Fuchsbandwurm und seinem Wirt von zentraler Rolle. Die Entschlüsselung des Echinococcus-Genoms gab Hinweise darauf, dass ein Mitglied der Tumornekrosefaktor-Rezeptor-Superfamilie, jedoch kein endogener TNF α ähnlicher Ligand im Genom kodiert wird. Ein Mitglied der TNFR-Superfamilie des Fuchsbandwurmes (EmTNFR) wurde in dieser Arbeit als membranständiger Rezeptor mit einer intrazellulären Todesdomäne (DD) und hoher Ähnlichkeit zum humanen Typ 16 der TNF-Rezeptor-Superfamilie, auch 〖p75〗^NTR genannt, charakterisiert. Sowohl in bioinformatischen als auch in Sequenzanalysen wurden drei alternative Splicing-Formen von emtnfr (emtnfr, emtnfr-v2 und emtnfr-v3) nachgewiesen. emtnfr-v2 entsteht durch Alternatives Splicing und kodiert ein Protein, das keine intrazelluläre Todesdomäne besitzt. emtnfr-v3 verwendet einen alternativen Transkriptionstart und wird von den letzten 3 Exons von emtnfr kodiert. emtnfr-v3, kodiert ein Protein ohne extrazelluläre Region, aber mit intrazellulärer Todesdomäne. Ein löslicher TNF-Rezeptor konnte auf Proteinebene nicht nachgewiesen werden. Aufgrund von phylogenetischen Analysen und der Rezeptor-Struktur ist zu vermuten, dass EmTNFR ein p75NTR Homolog ist und damit der ursprünglichen Form der TNF-Rezeptoren entspricht. Mitglieder eines intrazellulären TNF-Signalweges wurden in bioinformatischen Analysen beim Fuchsbandwurm E. multilocularis identifiziert. Expressionsuntersuchungen zeigten sowohl in Trankriptomdaten als auch auf Proteinebene eine starke Expression von EmTNFR in Primärzellen und im Metazestoden (MZ), dem pathogenen Stadium für den Zwischenwirt. Echinococcus-Stammzellkulturen zeigten nach RNA-Interferenz-basiertem Knockdown des EmTNFR-kodierenden Gens deutliche Entwicklungsdefekte. Des Weiteren zeigten Echinococcus-Stammzellkulturen nach einer Behandlung mit TNF-α, einem potentiellen Liganden des TNF-Rezeptors und einem zentralen Zytokin in der Immunabwehr des Zwischenwirtes, Entwicklungsfortschritte, wie eine verbesserte Bildung von MZ aus Stammzellen. Zusätzlich wurde in whole-mount in situ Hybridisierungs-Versuchen eine ubiquitäre Expression von emtnfr in der Germinalschicht des MZ sowie eine Spezifität von emtnfr für den MZ, welcher ursächlich für die AE ist, nachgewiesen. Somit scheinen sowohl EmTNFR als auch TNF-α eine wichtige Funktion bei der Entwicklung und Etablierung des Fuchsbandwurmes während der frühen Phase der Infektion des Zwischenwirtes zu haben. TNF-α könnte ein weiterer Faktor für den ausgeprägten Organtropismus des Parasiten zur Leber sein, denn dort bestehen durch Kupfferzellen produzierte hohe lokale Konzentration von TNF-α. Zusammenfassend deuten die hier erarbeiteten Daten darauf hin, dass EmTNFR über die Bindung von Wirts-TNF-α bei der frühen Entwicklung des Echincoccus-Metazestoden eine Rolle spielt. N2 - Alveolar echinococcosis (AE), which is caused by the metacestode larval stage of the fox tapeworm Echinococcus multilocularis, is a rare but severe, often fatal disease. Due to late diagnosis and advanced spread of the infection curative therapy is often not possible and the only treatment option is benzimidazole chemotherapy, which often must be taken lifelong and has adverse side effects. Improvement of therapeutic options is thus urgently needed. To this end, a closer understanding of parasite biology and communication mechanisms between parasite and host are helpful. In this work, focus was laid on the possibility of host-parasite cross-communication involving an evolutionarily conserved signalling pathway. By mining the Echinococcus genome sequence, a gene encoding a member of the tumor necrosis-factor-receptor family (TNF-R), was identified. In this work, EmTNFR, a member of the TNF-R superfamily, of the fox tapeworm was identified as a membrane bound receptor with intracellular death domain and highest similarity to human TNFRSF 16, also called p75NTR. In in silico analysis and cDNA sequencing, 3 alternative splice forms of emtnfr (emtnfr-v1, -v2 and -v3) were found. emtnfr-v2 is the result of alternative splicing and encodes a protein lacking the intracellular death domain. emtnfr-v3 employs an alternative transcription start and is encoded by the last 3 exons of emtnfr. emtnfr-v3 encodes a protein without extracellular domain, but containing an intracellular death domain. A soluble TNF-receptor could not be found in proteomic analysis. Based on phylogenetic analysis and receptor structure, EmTNFR is thought to be a homolog of p75NTR, corresponding to the ancient form of TNF receptors. Members of an intracellular TNF signaling pathway were identified in bioinformatic analyses in the fox tapeworm E. multilocularis, indicating the presence of a full TNFR signalling pathway. Expression studies showed in transcriptome data and at protein level a strong expression of EmTNFR in primary cells and in the metacestode (MZ), the pathogenic stage for the intermediate host. Echinococcus stem cell cultures showed marked developmental defects after RNAi based knockdown of the EmTNFR-encoding gene. Furthermore, Echinococcus stem cell culture displayed accelerated developmental progress such as enhanced formation of MZ from stem cells after treatment with TNF-α, a potential ligand of the TNF receptor, and a central cytokine in the immune defense of the intermediate host. In addition, whole-mount in situ hybridization experiments demonstrated ubiquitous expression of emtnfr in the germinal layer of MZ and specificity of emtnfr for MZ, the causative agent of AE. Thus, both EmTNFR and TNF-α appear to have an important function in development and establishment of the fox tapeworm during the early phase of infection of the intermediate host. TNF-α could be an additional factor for the pronounced organ tropism of the parasite to the liver, caused by a high local concentration of TNF-α produced by Kupffer cells. In summary, the data generated in this work suggest that EmTNFR plays a role in the early development of Echinococcus metacestode via binding of host TNF-α. KW - Fuchsbandwurm KW - Wirt-Parasit-Beziehung KW - Parasit KW - Tumor-Nekrose-Faktor KW - Echinococcus multilocularis KW - TNF-Rezeptor KW - Wirt-Parasiten-Interaktion KW - Molekulare Charakterisierung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369397 ER - TY - THES A1 - Reissland, Michaela T1 - USP10 is a \(de\) \(novo\) tumour-specific regulator of β-Catenin and contributes to cancer stem cell maintenance and tumour progression T1 - USP10 ist ein \(de\) \(novo\) tumorspezifischer Regulator von ß-Catenin und trägt zur Erhaltung von Krebsstammzellen und zur Tumorprogression bei N2 - Colorectal Cancer (CRC) is the third most common cancer in the US. The majority of CRC cases are due to deregulated WNT-signalling pathway. These alterations are mainly caused by mutations in the tumour suppressor gene APC or in CTNNB1, encoding the key effector protein of this pathway, β-Catenin. In canonical WNT-signalling, β-Catenin activates the transcription of several target genes, encoding for proteins involved in proliferation, such as MYC, JUN and NOTCH. Being such a critical regulator of these proto-oncogenes, the stability of β-Catenin is tightly regulated by the Ubiquitin-Proteasome System. Several E3 ligases that ubiquitylate and degrade β-Catenin have been described in the past, but the antagonists, the deubiquitylases, are still unknown. By performing an unbiased siRNA screen, the deubiquitylase USP10 was identified as a de novo positive regulator of β-Catenin stability in CRC derived cells. USP10 has previously been shown in the literature to regulate both mutant and wild type TP53 stability, to deubiquitylate NOTCH1 in endothelial cells and to be involved in the regulation of AMPKα signalling. Overall, however, its role in colorectal tumorigenesis remains controversial. By analysing publicly available protein and gene expression data from colorectal cancer patients, we have shown that USP10 is strongly upregulated or amplified upon transformation and that its expression correlates positively with CTNNB1 expression. In contrast, basal USP10 levels were found in non-transformed tissues, but surprisingly USP10 is upregulated in intestinal stem cells. Endogenous interaction studies in CRC-derived cell lines, with different extend of APCtruncation, revealed an APC-dependent mode of action for both proteins. Furthermore, by utilising CRISPR/Cas9, shRNA-mediated knock-down and overexpression of USP10, we could demonstrate a regulation of β-Catenin stability by USP10 in CRC cell lines. It is widely excepted that 2D cell culture systems do not reflect complexity, architecture and heterogeneity and are therefore not suitable to answer complex biological questions. To overcome this, we established the isolation, cultivation and genetically modification of murine intestinal organoids and utilised this system to study Usp10s role ex vivo. By performing RNA sequencing, dependent on different Usp10 levels, we were able to recapitulate the previous findings and demonstrated Usp10 as important regulator of β-dependent regulation of stem cell homeostasis. Since genetic depletion of USP10 resulted in down-regulation of β-Catenin-dependent transcription, therapeutic intervention of USP10 in colorectal cancer was also investigated. Commercial and newly developed inhibitors were tested for their efficacy against USP10, but failed to significantly inhibit USP10 activity in colorectal cancer cells. To validate the findings from this work also in vivo, development of a novel mouse model for colorectal cancer has begun. By combining CRISPR/Cas9 and classical genetic engineering with viral injection strategies, WT and genetically modified mice could be transformed and, at least in some animals, intestinal lesions were detectable at the microscopic level. The inhibition of USP10, which we could describe as a de novo tumour-specific regulator of β-Catenin, could become a new therapeutic strategy for colorectal cancer patients. N2 - Darmkrebs ist die dritthäufigste Krebsart in den USA. Die Mehrheit der Darmkrebsfälle sind auf einen deregulierten WNT-Signalweg zurückzuführen. Diese Veränderungen wer- den hauptsächlich durch Mutationen im Tumorsuppressor-Gen APC oder in CTNNB1 verursacht, welches für das zentrale Protein dieses Signalwegs, β-Catenin, kodiert. Beim kanonischen WNT-Signalweg aktiviert β-Catenin die Transkription mehrerer Gene, die für, an der Proliferation beteiligte Proteine wie MYC, JUN und NOTCH, kodieren. Da β-Catenin ein kritischer Regulator dieser proto-Onkogene ist, wird die Stabilität von β-Catenin durch das Ubiquitin-Proteasom-System streng reguliert. In der Vergangen- heit wurden mehrere E3-Ligasen beschrieben, die β-Catenin ubiquitylieren und abbauen, aber die Deubiquitylasen, sind grö𐀀tenteils noch unbekannt. Mit Hilfe eines unvoreingenommenen siRNA-Screens wurde die Deubiquitylase USP10 als de novo Regulator der β-Catenin-Stabilität in Darmkrebs-Zellen identifiziert. In der Literatur wurde bereits gezeigt, dass USP10 sowohl die Stabilität von mutiertem als auch von wild typ TP53 reguliert, NOTCH1 in Endothelzellen deubiquityliert und an der Regulation des AMPKα Signalwegs beteiligt ist. Insgesamt bleibt seine Rolle in der kolorektalen Tumorgenese aber bisher umstritten. Anhand der Analyse öffentlich zugänglicher Protein- und Genexpressionsdaten haben wir gezeigt, dass USP10 bei der Transformation stark hochreguliert oder amplifiziert wird und dass seine Expression positiv mit der von CTNNB1 korreliert. Im Gegensatz dazu wurden in nicht transformiertem Gewebe basale USP10-Spiegel gefunden, aber überraschenderweise ist USP10 in intestinalen Stammzellen hochreguliert. Endogene Interaktionsstudien in Darmkrebs-Zelllinien mit unterschiedlichem Ausma𐀀 an APC-Trunkierung zeigten eine APC-abhängige Interaktion für beide Proteine. Darüber hinaus konnten wir mit Hilfe von CRISPR/Cas9, shRNA-vermitteltem Knock-down und Überexpression von USP10 eine Regulation der β-Catenin-Stabilität durch USP10 in Darmkrebs-Zelllinien nachweisen. Es ist allgemein bekannt, dass 2D-Zellkultursysteme die Komplexität, Architektur und Heterogenität nicht widerspiegeln und daher nicht geeignet sind, um komplexe biologische Fragen zu beantworten. Um dies zu überwinden, haben wir die Isolierung, Kultivierung und genetische Veränderung von murinen Dar- morganoiden etabliert und dieses System genutzt, um die Rolle von Usp10 ex vivo zu untersuchen. Durch die Durchführung von RNA-Sequenzierungen in Abhängigkeit von unterschiedlichen Usp10-Spiegeln konnten wir die bisherigen Ergebnisse rekapitulieren und Usp10 als wichtigen Regulator der β-Catenin-abhängigen Regulation der Stammzell- homöostase nachweisen. Da die genetische Depletion von USP10 zu einer Herunterregulierung der β-Catenin- abhängigen Transkription führte, wurde auch die therapeutische Intervention von USP10 in Darmkrebs untersucht. Kommerzielle und neu entwickelte Inhibitoren wurden auf ihre Wirksamkeit gegen USP10 getestet, konnten jedoch die Aktivität von USP10 in Darmkrebs- Zellen nicht hemmen. Um die Erkenntnisse aus dieser Arbeit auch in vivo zu validieren, wurde mit der Entwicklung eines neuartigen Mausmodells für Darmkrebs begonnen. Durch die Kombination von CRISPR/Cas9 und klassischer Gentechnik mit viralen Injektionsstrategien konnten WT- und gentechnisch veränderte Mäuse trans- formiert werden und zumindest bei einigen Tieren waren Darmläsionen auf mikroskopis- cher Ebene nachweisbar. Die Inhibtierung von USP10, als de novo tumorspezifischer Regulator von β-Catenin, könnte eine neue therapeutische Strategie für Darmkrebs-Patienten werden. KW - Biomedizin KW - Biomedicine Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319579 ER - TY - JOUR A1 - Morimoto, Yoshiro A1 - Shimada-Sugimoto, Mihoko A1 - Otowa, Takeshi A1 - Yoshida, Shintaro A1 - Kinoshita, Akira A1 - Mishima, Hiroyuki A1 - Yamaguchi, Naohiro A1 - Mori, Takatoshi A1 - Imamura, Akira A1 - Ozawa, Hiroki A1 - Kurotaki, Naohiro A1 - Ziegler, Christiane A1 - Domschke, Katharina A1 - Deckert, Jürgen A1 - Umekage, Tadashi A1 - Tochigi, Mamoru A1 - Kaiya, Hisanobu A1 - Okazaki, Yuji A1 - Tokunaga, Katsushi A1 - Sasaki, Tsukasa A1 - Yoshiura, Koh-ichiro A1 - Ono, Shinji T1 - Whole-exome sequencing and gene-based rare variant association tests suggest that PLA2G4E might be a risk gene for panic disorder JF - Translational Psychiatry N2 - Panic disorder (PD) is characterized by recurrent and unexpected panic attacks, subsequent anticipatory anxiety, and phobic avoidance. Recent epidemiological and genetic studies have revealed that genetic factors contribute to the pathogenesis of PD. We performed whole-exome sequencing on one Japanese family, including multiple patients with panic disorder, which identified seven rare protein-altering variants. We then screened these genes in a Japanese PD case–control group (384 sporadic PD patients and 571 controls), resulting in the detection of three novel single nucleotide variants as potential candidates for PD (chr15: 42631993, T>C in GANC; chr15: 42342861, G>T in PLA2G4E; chr20: 3641457, G>C in GFRA4). Statistical analyses of these three genes showed that PLA2G4E yielded the lowest p value in gene-based rare variant association tests by Efficient and Parallelizable Association Container Toolbox algorithms; however, the p value did not reach the significance threshold in the Japanese. Likewise, in a German case–control study (96 sporadic PD patients and 96 controls), PLA2G4E showed the lowest p value but again did not reach the significance threshold. In conclusion, we failed to find any significant variants or genes responsible for the development of PD. Nonetheless, our results still leave open the possibility that rare protein-altering variants in PLA2G4E contribute to the risk of PD, considering the function of this gene. KW - clinical genetics KW - medical genetics Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224192 VL - 8 ER - TY - JOUR A1 - Nerreter, Thomas A1 - Letschert, Sebastian A1 - Götz, Ralph A1 - Doose, Sören A1 - Danhof, Sophia A1 - Einsele, Hermann A1 - Sauer, Markus A1 - Hudecek, Michael T1 - Super-resolution microscopy reveals ultra-low CD19 expression on myeloma cells that triggers elimination by CD19 CAR-T JF - Nature Communications N2 - Immunotherapy with chimeric antigen receptor-engineered T-cells (CAR-T) is under investigation in multiple myeloma. There are reports of myeloma remission after CD19 CAR-T therapy, although CD19 is hardly detectable on myeloma cells by flow cytometry (FC). We apply single molecule-sensitive direct stochastic optical reconstruction microscopy (dSTORM), and demonstrate CD19 expression on a fraction of myeloma cells (10.3–80%) in 10 out of 14 patients (density: 13–5,000 molecules per cell). In contrast, FC detects CD19 in only 2 of these 10 patients, on a smaller fraction of cells. Treatment with CD19 CAR-T in vitro results in elimination of CD19-positive myeloma cells, including those with <100 CD19 molecules per cell. Similar data are obtained by dSTORM analyses of CD20 expression on myeloma cells and CD20 CAR-T. These data establish a sensitivity threshold for CAR-T and illustrate how super-resolution microscopy can guide patient selection in immunotherapy to exploit ultra-low density antigens. KW - cancer imaging KW - cancer immunotherapy KW - imaging Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232258 VL - 10 ER - TY - JOUR A1 - Odin, Per A1 - Chaudhuri, K. Ray A1 - Volkmann, Jens A1 - Antonini, Angelo A1 - Storch, Alexander A1 - Dietrichs, Espen A1 - Pirtošek, Zvezdan A1 - Henriksen, Tove A1 - Horne, Malcolm A1 - Devos, David A1 - Bergquist, Filip T1 - Viewpoint and practical recommendations from a movement disorder specialist panel on objective measurement in the clinical management of Parkinson’s disease JF - npj Parkinson's Disease N2 - Motor aspects of Parkinson’s disease, such as fluctuations and dyskinesia, can be reliably evaluated using a variety of “wearable” technologies, but practical guidance on objective measurement (OM) and the optimum use of these devices is lacking. Therefore, as a first step, a panel of movement disorder specialists met to provide guidance on how OM could be assessed and incorporated into clinical guidelines. A key aspect of the incorporation of OM into the management of Parkinson’s disease (PD) is defining cutoff values that separate “controlled” from “uncontrolled” symptoms that can be modified by therapy and that relate to an outcome that is relevant to the person with PD (such as quality of life). Defining cutoffs by consensus, which can be subsequently tested and refined, is the first step to optimizing OM in the management of PD. OM should be used by all clinicians that treat people with PD but the least experienced may find the most value, but this requires guidance from experts to allow non-experts to apply guidelines. While evidence is gained for devices that produce OM, expert opinion is needed to supplement the evidence base. KW - Parkinson's disease Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234435 VL - 4 ER - TY - JOUR A1 - Scholz, S. L. A1 - Cosgarea, I. A1 - Süßkind, D. A1 - Murali, R. A1 - Möller, I. A1 - Reis, H. A1 - Leonardelli, S. A1 - Schilling, B. A1 - Schimming, T. A1 - Hadaschik, E. A1 - Franklin, C. A1 - Paschen, A. A1 - Sucker, A. A1 - Steuhl, K. P. A1 - Schadendorf, D. A1 - Westekemper, H. A1 - Griewank, K. G. T1 - NF1 mutations in conjunctival melanoma JF - British Journal of Cancer N2 - Background Conjunctival melanoma is a potentially deadly eye tumour. Despite effective local therapies, tumour recurrence and metastasis remain frequent. The genetics of conjunctival melanomas remain incompletely understood. Methods A large cohort of 63 conjunctival melanomas was screened for gene mutations known to be important in other melanoma subtypes by targeted next-generation sequencing. Mutation status was correlated with patient prognosis. Results Frequent mutations in genes activating the MAP kinase pathway were identified. NF1 mutations were most frequent (n = 21, 33%). Recurrent activating mutations were also identified in BRAF (n = 16, 25%) and RAS genes (n = 12, 19%; 11 NRAS and 1 KRAS). Conclusions Similar to cutaneous melanomas, conjunctival melanomas can be grouped genetically into four groups: BRAF-mutated, RAS-mutated, NF1-mutated and triple wild-type melanomas. This genetic classification may be useful for assessment of therapeutic options for patients with metastatic conjunctival melanoma KW - cancer genetics KW - eye cancer Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233329 VL - 118 ER - TY - JOUR A1 - Bruchhagen, Christin A1 - Jarick, Marcel A1 - Mewis, Carolin A1 - Hertlein, Tobias A1 - Niemann, Silke A1 - Ohlsen, Knut A1 - Peters, Georg A1 - Planz, Oliver A1 - Ludwig, Stephan A1 - Ehrhardt, Christina T1 - Metabolic conversion of CI-1040 turns a cellular MEK-inhibitor into an antibacterial compound JF - Scientific Reports N2 - Influenza virus (IV) infections cause severe respiratory illnesses that can be complicated by bacterial super-infections. Previously, we identified the cellular Raf-MEK-ERK cascade as a promising antiviral target. Inhibitors of MEK, such as CI-1040, showed potent antiviral activity. However, it remained unclear if this inhibitor and its active form, ATR-002, might sensitize host cells to either IV or secondary bacterial infections. To address these questions, we studied the anti-pathogen activity of ATR-002 in comparison to CI-1040, particularly, its impact on Staphylococcus aureus (S. aureus), which is a major cause of IV super-infections. We analysed IV and S. aureus titres in vitro during super-infection in the presence and absence of the drugs and characterized the direct impact of ATR-002 on bacterial growth and phenotypic changes. Importantly, neither CI-1040 nor ATR-002 treatment led to increased bacterial titres during super-infection, indicating that the drug does not sensitize cells for bacterial infection. In contrast, we rather observed reduced bacterial titres in presence of ATR-002. Surprisingly, ATR-002 also led to reduced bacterial growth in suspension cultures, reduced stress- and antibiotic tolerance without resistance induction. Our data identified for the first time that a particular MEK-inhibitor metabolite exhibits direct antibacterial activity, which is likely due to interference with the bacterial PknB kinase/Stp phosphatase signalling system. KW - antimicrobials KW - pathogens Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221648 VL - 8 ER - TY - JOUR A1 - Castilho, Miguel A1 - Hochleitner, Gernot A1 - Wilson, Wouter A1 - van Rietbergen, Bert A1 - Dalton, Paul D. A1 - Groll, Jürgen A1 - Malda, Jos A1 - Ito, Keita T1 - Mechanical behavior of a soft hydrogel reinforced with three-dimensional printed microfibre scaffolds JF - Scientific Reports N2 - Reinforcing hydrogels with micro-fibre scaffolds obtained by a Melt-Electrospinning Writing (MEW) process has demonstrated great promise for developing tissue engineered (TE) constructs with mechanical properties compatible to native tissues. However, the mechanical performance and reinforcement mechanism of the micro-fibre reinforced hydrogels is not yet fully understood. In this study, FE models, implementing material properties measured experimentally, were used to explore the reinforcement mechanism of fibre-hydrogel composites. First, a continuum FE model based on idealized scaffold geometry was used to capture reinforcement effects related to the suppression of lateral gel expansion by the scaffold, while a second micro-FE model based on micro-CT images of the real construct geometry during compaction captured the effects of load transfer through the scaffold interconnections. Results demonstrate that the reinforcement mechanism at higher scaffold volume fractions was dominated by the load carrying-ability of the fibre scaffold interconnections, which was much higher than expected based on testing scaffolds alone because the hydrogel provides resistance against buckling of the scaffold. We propose that the theoretical understanding presented in this work will assist the design of more effective composite constructs with potential applications in a wide range of TE conditions. KW - biomedical engineering KW - biomedical materials KW - gels and hydrogels Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222280 VL - 8 ER - TY - JOUR A1 - Al-Zaben, Naim A1 - Medyukhina, Anna A1 - Dietrich, Stefanie A1 - Marolda, Alessandra A1 - Hünniger, Kerstin A1 - Kurzai, Oliver A1 - Figge, Marc Thilo T1 - Automated tracking of label-free cells with enhanced recognition of whole tracks JF - Scientific Reports N2 - Migration and interactions of immune cells are routinely studied by time-lapse microscopy of in vitro migration and confrontation assays. To objectively quantify the dynamic behavior of cells, software tools for automated cell tracking can be applied. However, many existing tracking algorithms recognize only rather short fragments of a whole cell track and rely on cell staining to enhance cell segmentation. While our previously developed segmentation approach enables tracking of label-free cells, it still suffers from frequently recognizing only short track fragments. In this study, we identify sources of track fragmentation and provide solutions to obtain longer cell tracks. This is achieved by improving the detection of low-contrast cells and by optimizing the value of the gap size parameter, which defines the number of missing cell positions between track fragments that is accepted for still connecting them into one track. We find that the enhanced track recognition increases the average length of cell tracks up to 2.2-fold. Recognizing cell tracks as a whole will enable studying and quantifying more complex patterns of cell behavior, e.g. switches in migration mode or dependence of the phagocytosis efficiency on the number and type of preceding interactions. Such quantitative analyses will improve our understanding of how immune cells interact and function in health and disease. KW - image processing KW - software Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221093 VL - 9 ER - TY - JOUR A1 - de Jong, Simone A1 - Diniz, Mateus Jose Abdalla A1 - Saloma, Andiara A1 - Gadelha, Ary A1 - Santoro, Marcos L. A1 - Ota, Vanessa K. A1 - Noto, Cristiano A1 - Curtis, Charles A1 - Newhouse, Stephen J. A1 - Patel, Hamel A1 - Hall, Lynsey S. A1 - O'Reilly, Paul F. A1 - Belangero, Sintia I. A1 - Bressan, Rodrigo A. A1 - Breen, Gerome T1 - Applying polygenic risk scoring for psychiatric disorders to a large family with bipolar disorder and major depressive disorder JF - Communications Biology N2 - Psychiatric disorders are thought to have a complex genetic pathology consisting of interplay of common and rare variation. Traditionally, pedigrees are used to shed light on the latter only, while here we discuss the application of polygenic risk scores to also highlight patterns of common genetic risk. We analyze polygenic risk scores for psychiatric disorders in a large pedigree (n ~ 260) in which 30% of family members suffer from major depressive disorder or bipolar disorder. Studying patterns of assortative mating and anticipation, it appears increased polygenic risk is contributed by affected individuals who married into the family, resulting in an increasing genetic risk over generations. This may explain the observation of anticipation in mood disorders, whereby onset is earlier and the severity increases over the generations of a family. Joint analyses of rare and common variation may be a powerful way to understand the familial genetics of psychiatric disorders. KW - bipolar disorder KW - depression KW - genetic association study KW - genetic linkage study Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223622 VL - 1 ER - TY - JOUR A1 - Dekker, Annelot M. A1 - Diekstra, Frank P. A1 - Pulit, Sara L. A1 - Tazelaar, Gijs H. P. A1 - van der Spek, Rick A. A1 - van Rheenen, Wouter A1 - van Eijk, Kristel R. A1 - Calvo, Andrea A1 - Brunetti, Maura A1 - Van Damme, Philip A1 - Robberecht, Wim A1 - Hardiman, Orla A1 - McLaughlin, Russell A1 - Chiò, Adriano A1 - Sendtner, Michael A1 - Ludolph, Albert C. A1 - Weishaupt, Jochen H. A1 - Pardina, Jesus S. Mora A1 - van den Berg, Leonard H. A1 - Veldink, Jan H. T1 - Exome array analysis of rare and low frequency variants in amyotrophic lateral sclerosis JF - Scientific Reports N2 - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that affects 1 in ~350 individuals. Genetic association studies have established ALS as a multifactorial disease with heritability estimated at ~61%, and recent studies show a prominent role for rare variation in its genetic architecture. To identify rare variants associated with disease onset we performed exome array genotyping in 4,244 cases and 3,106 controls from European cohorts. In this largest exome-wide study of rare variants in ALS to date, we performed single-variant association testing, gene-based burden, and exome-wide individual set-unique burden (ISUB) testing to identify single or aggregated rare variation that modifies disease risk. In single-variant testing no variants reached exome-wide significance, likely due to limited statistical power. Gene-based burden testing of rare non-synonymous and loss-of-function variants showed NEK1 as the top associated gene. ISUB analysis did not show an increased exome-wide burden of deleterious variants in patients, possibly suggesting a more region-specific role for rare variation. Complete summary statistics are released publicly. This study did not implicate new risk loci, emphasizing the immediate need for future large-scale collaborations in ALS that will expand available sample sizes, increase genome coverage, and improve our ability to detect rare variants associated to ALS. KW - amyotrophic lateral sclerosis KW - genome-wide association studies Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223686 VL - 9 ER -