@article{GhoshHoenscheidDueckersetal.2017, author = {Ghosh, Sujal and H{\"o}nscheid, Andrea and D{\"u}ckers, Gregor and Ginzel, Sebastian and Gohlke, Holger and Gombert, Michael and Kempkes, Bettina and Klapper, Wolfram and Kuhlen, Michaela and Laws, Hans-J{\"u}rgen and Linka, Ren{\´e} Martin and Meisel, Roland and Mielke, Christian and Niehues, Tim and Schindler, Detlev and Schneider, Dominik and Schuster, Friedhelm R. and Speckmann, Carsten and Borkhardt, Arndt}, title = {Human RAD52 - a novel player in DNA repair in cancer and immunodeficiency}, series = {Haematologica}, volume = {102}, journal = {Haematologica}, number = {2}, doi = {10.3324/haematol.2016.155838}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-180862}, pages = {e69-e72}, year = {2017}, abstract = {No abstract available.}, language = {en} } @article{HernandezJoseRamirezMinguillonetal.2018, author = {Hern{\´a}ndez, Gonzalo and Jos{\´e} Ram{\´i}rez, Mar{\´i}a and Minguill{\´o}n, Jordi and Quiles, Paco and Ruiz de Garibay, Gorka and Aza-Carmona, Miriam and Bogliolo, Massimo and Pujol, Roser and Prados-Carvajal, Rosario and Fern{\´a}ndez, Juana and Garc{\´i}a, Nadia and L{\´o}pez, Adri{\`a} and Guti{\´e}rrez-Enr{\´i}quez, Sara and Diez, Orland and Ben{\´i}tez, Javier and Salinas, M{\´o}nica and Teul{\´e}, Alex and Brunet, Joan and Radice, Paolo and Peterlongo, Paolo and Schindler, Detlev and Huertas, Pablo and Puente, Xose S. and L{\´a}zaro, Conxi and {\`A}ngel Pujana, Miquel and Surrall{\´e}s, Jordi}, title = {Decapping protein EDC4 regulates DNA repair and phenocopies BRCA1}, series = {Nature Communications}, volume = {9}, journal = {Nature Communications}, doi = {10.1038/s41467-018-03433-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-319929}, year = {2018}, abstract = {BRCA1 is a tumor suppressor that regulates DNA repair by homologous recombination. Germline mutations in BRCA1 are associated with increased risk of breast and ovarian cancer and BRCA1 deficient tumors are exquisitely sensitive to poly (ADP-ribose) polymerase (PARP) inhibitors. Therefore, uncovering additional components of this DNA repair pathway is of extreme importance for further understanding cancer development and therapeutic vulnerabilities. Here, we identify EDC4, a known component of processing-bodies and regulator of mRNA decapping, as a member of the BRCA1-BRIP1-TOPBP1 complex. EDC4 plays a key role in homologous recombination by stimulating end resection at double-strand breaks. EDC4 deficiency leads to genome instability and hypersensitivity to DNA interstrand cross-linking drugs and PARP inhibitors. Lack-of-function mutations in EDC4 were detected in BRCA1/2-mutation-negative breast cancer cases, suggesting a role in breast cancer susceptibility. Collectively, this study recognizes EDC4 with a dual role in decapping and DNA repair whose inactivation phenocopies BRCA1 deficiency.}, language = {en} } @phdthesis{Hahn2024, author = {Hahn, Sarah}, title = {Investigating non-canonical, 5' UTR-dependent translation of MYC and its impact on colorectal cancer development}, doi = {10.25972/OPUS-36420}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-364202}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Colorectal cancer (CRC) is the second most common tumour disease in Germany, with the sequential accumulation of certain mutations playing a decisive role in the transition from adenoma to carcinoma. In particular, deregulation of the Wnt signalling pathway and the associated deregulated expression of the MYC oncoprotein play a crucial role. Targeting MYC thus represents an important therapeutic approach in the treatment of tumours. Since direct inhibition of MYC is challenging, various approaches have been pursued to date to target MYC indirectly. The MYC 5' UTR contains an internal ribosomal entry site (IRES), which has a particular role in the initiation of MYC translation, especially in multiple myeloma. As basis for this work, it was hypothesised on the basis of previous data that translation of MYC potentially occurs via its IRES in CRC as well. Based on this, two IRES inhibitors were tested for their potential to regulate MYC expression in CRC cells. In addition, alternative, 5' UTR-dependent translation of MYC and interacting factors were investigated. EIF3D was identified as a MYC 5' UTR binding protein which has the potential to regulate MYC expression in CRC. The results of this work suggest that there is a link between eIF3D and MYC expression/translation, rendering eIF3D a potential therapeutic target for MYC-driven CRCs.}, subject = {Myc}, language = {en} } @phdthesis{Yu2024, author = {Yu, Yanying}, title = {Applied machine learning for the analysis of CRISPR-Cas systems}, doi = {10.25972/OPUS-32021}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-320219}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Among the defense strategies developed in microbes over millions of years, the innate adaptive CRISPR-Cas immune systems have spread across most of bacteria and archaea. The flexibility, simplicity, and specificity of CRISPR-Cas systems have laid the foundation for CRISPR-based genetic tools. Yet, the efficient administration of CRISPR-based tools demands rational designs to maximize the on-target efficiency and off-target specificity. Specifically, the selection of guide RNAs (gRNAs), which play a crucial role in the target recognition of CRISPR-Cas systems, is non-trivial. Despite the fact that the emerging machine learning techniques provide a solution to aid in gRNA design with prediction algorithms, design rules for many CRISPR-Cas systems are ill-defined, hindering their broader applications. CRISPR interference (CRISPRi), an alternative gene silencing technique using a catalytically dead Cas protein to interfere with transcription, is a leading technique in bacteria for functional interrogation, pathway manipulation, and genome-wide screens. Although the application is promising, it also is hindered by under-investigated design rules. Therefore, in this work, I develop a state-of-art predictive machine learning model for guide silencing efficiency in bacteria leveraging the advantages of feature engineering, data integration, interpretable AI, and automated machine learning. I first systematically investigate the influential factors that attribute to the extent of depletion in multiple CRISPRi genome-wide essentiality screens in Escherichia coli and demonstrate the surprising dominant contribution of gene-specific effects, such as gene expression level. These observations allowed me to segregate the confounding gene-specific effects using a mixed-effect random forest (MERF) model to provide a better estimate of guide efficiency, together with the improvement led by integrating multiple screens. The MERF model outperformed existing tools in an independent high-throughput saturating screen. I next interpret the predictive model to extract the design rules for robust gene silencing, such as the preference for cytosine and disfavoring for guanine and thymine within and around the protospacer adjacent motif (PAM) sequence. I further incorporated the MERF model in a web-based tool that is freely accessible at www.ciao.helmholtz-hiri.de. When comparing the MERF model with existing tools, the performance of the alternative gRNA design tool optimized for CRISPRi in eukaryotes when applied to bacteria was far from satisfying, questioning the robustness of prediction algorithms across organisms. In addition, the CRISPR-Cas systems exhibit diverse mechanisms albeit with some similarities. The captured predictive patterns from one dataset thereby are at risk of poor generalization when applied across organisms and CRISPR-Cas techniques. To fill the gap, the machine learning approach I present here for CRISPRi could serve as a blueprint for the effective development of prediction algorithms for specific organisms or CRISPR-Cas systems of interest. The explicit workflow includes three principle steps: 1) accommodating the feature set for the CRISPR-Cas system or technique; 2) optimizing a machine learning model using automated machine learning; 3) explaining the model using interpretable AI. To illustrate the applicability of the workflow and diversity of results when applied across different bacteria and CRISPR-Cas systems, I have applied this workflow to analyze three distinct CRISPR-Cas genome-wide screens. From the CRISPR base editor essentiality screen in E. coli, I have determined the PAM preference and sequence context in the editing window for efficient editing, such as A at the 2nd position of PAM, A/TT/TG downstream of PAM, and TC at the 4th to 5th position of gRNAs. From the CRISPR-Cas13a screen in E. coli, in addition to the strong correlation with the guide depletion, the target expression level is the strongest predictor in the model, supporting it as a main determinant of the activation of Cas13-induced immunity and better characterizing the CRISPR-Cas13 system. From the CRISPR-Cas12a screen in Klebsiella pneumoniae, I have extracted the design rules for robust antimicrobial activity across K. pneumoniae strains and provided a predictive algorithm for gRNA design, facilitating CRISPR-Cas12a as an alternative technique to tackle antibiotic resistance. Overall, this thesis presents an accurate prediction algorithm for CRISPRi guide efficiency in bacteria, providing insights into the determinants of efficient silencing and guide designs. The systematic exploration has led to a robust machine learning approach for effective model development in other bacteria and CRISPR-Cas systems. Applying the approach in the analysis of independent CRISPR-Cas screens not only sheds light on the design rules but also the mechanisms of the CRISPR-Cas systems. Together, I demonstrate that applied machine learning paves the way to a deeper understanding and a broader application of CRISPR-Cas systems.}, subject = {Maschinelles Lernen}, language = {en} } @article{Gohla2019, author = {Gohla, Antje}, title = {Do metabolic HAD phosphatases moonlight as protein phosphatases?}, series = {BBA - Molecular Cell Research}, volume = {1866}, journal = {BBA - Molecular Cell Research}, doi = {10.1016/j.bbamcr.2018.07.007}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233168}, pages = {153-166}, year = {2019}, abstract = {Mammalian haloacid dehalogenase (HAD)-type phosphatases have evolved to dephosphorylate a wide range of small metabolites, but can also target macromolecules such as serine/threonine, tyrosine-, and histidine-phosphorylated proteins. To accomplish these tasks, HAD phosphatases are equipped with cap domains that control access to the active site and provide substrate specificity determinants. A number of capped HAD phosphatases impact protein phosphorylation, although structural data are consistent with small metabolite substrates rather than protein substrates. This review discusses the structures, functions and disease implications of the three closely related, capped HAD phosphatases pyridoxal phosphatase (PDXP or chronophin), phosphoglycolate phosphatase (PGP, also termed AUM or glycerol phosphatase) and phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP or HDHD2B). Evidence in support of small metabolite and protein phosphatase activity is discussed in the context of the diversity of their biological functions.}, language = {en} } @article{CasarottoTurcoComanduccietal.2019, author = {Casarotto, Silvia and Turco, Francesco and Comanducci, Angela and Perretti, Alessio and Marotta, Giorgio and Pezzoli, Gianni and Rosanova, Mario and Isaias, Ioannis U.}, title = {Excitability of the supplementary motor area in Parkinson's disease depends on subcortical damage}, series = {Brain Stimulation}, volume = {12}, journal = {Brain Stimulation}, doi = {10.1016/j.brs.2018.10.011}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-222261}, pages = {152-160}, year = {2019}, abstract = {Background Cortical dysfunctioning significantly contributes to the pathogenesis of motor symptoms in Parkinson's disease (PD). Objective We aimed at testing whether an acute levodopa administration has measurable and specific cortical effects possibly related to striatal dopaminergic deficit. Methods In thirteen PD patients, we measured the electroencephalographic responses to transcranial magnetic stimulation (TMS/EEG) of the supplementary motor area and superior parietal lobule (n = 8) before and after an acute intake of levodopa. We also performed a single-photon emission computed tomography and [123I]N-ω-fluoropropyl-2β-carbomethoxy-3β-(4-iodophenyl)nortropane to identify the more affected and the less affected brain side in each patient, according to the dopaminergic innervation loss of the putamen. Cortical excitability changes before and after an acute intake of levodopa were computed and compared between the more and the less affected brain side at the single-patient as well as at the group level. Results We found that levodopa intake induces a significant increase (P < 0.01) of cortical excitability nearby the supplementary motor area in the more affected brain side, greater (P < 0.025) than in the less affected brain side. Notably, cortical excitability changes nearby the superior parietal lobule were not statistically significant. Conclusions These results strengthen the idea that dysfunction of specific cortico-subcortical circuits may contribute to pathophysiology of PD symptoms. Most important, they support the use of navigated TMS/EEG as a non-invasive tool to better understand the pathophysiology of PD.}, language = {en} } @article{EisenhoferPeitzschKadenetal.2019, author = {Eisenhofer, Graeme and Peitzsch, Mirko and Kaden, Denise and Langton, Katharina and Mangelis, Anastasios and Pamporaki, Christina and Masjkur, Jimmy and Geroula, Aikaterini and Kurlbaum, Max and Deutschbein, Timo and Beuschlein, Felix and Prejbisz, Aleksander and Bornstein, Stefan R. and Lenders, Jacques W. M.}, title = {Reference intervals for LC-MS/MS measurements of plasma free, urinary free and urinary acid-hydrolyzed deconjugated normetanephrine, metanephrine and methoxytyramine}, series = {Clinica Chimica Acta}, volume = {490}, journal = {Clinica Chimica Acta}, doi = {10.1016/j.cca.2018.12.019}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-226598}, pages = {46-54}, year = {2019}, abstract = {Background Plasma or urinary metanephrines are recommended for screening of pheochromocytomas and paragangliomas (PPGLs). Measurements of urinary free rather than deconjugated metanephrines and additional measurements of methoxytyramine represent other developments. For all measurements there is need for reference intervals. Methods Plasma free, urinary free and urinary deconjugated O-methylated catecholamine metabolites were measured by LC-MS/MS in specimens from 590 hypertensives and normotensives. Reference intervals were optimized using data from 2,056 patients tested for PPGLs. Results Multivariate analyses, correcting for age and body surface area, indicated higher plasma and urinary metanephrine in males than females and sex differences in urinary normetanephrine and free methoxytyramine that largely reflected body size variation. There were positive associations of age with plasma metabolites, but negative relationships with urinary free metanephrine and methoxytyramine. Plasma and urinary normetanephrine were higher in hypertensives than normotensives, but differences were small. Optimization of reference intervals using the data from patients tested for PPGLs indicated that age was the most important consideration for plasma normetanephrine and sex most practical for urinary metabolites. Conclusion This study clarifies impacts of demographic and anthropometric variables on catecholamine metabolites, verifies use of age-specific reference intervals for plasma normetanephrine and establishes sex-specific reference intervals for urinary metabolites.}, language = {en} } @article{LieseSchoenvanderLindenetal.2019, author = {Liese, J. G. and Schoen, C. and van der Linden, M. and Lehmann, L. and Goettler, D. and Keller, S. and Maier, A. and Segerer, F. and Rose, M. A. and Streng, A.}, title = {Changes in the incidence and bacterial aetiology of paediatric parapneumonic pleural effusions/empyema in Germany, 2010-2017: a nationwide surveillance study}, series = {Clinical Microbiology and Infection}, volume = {25}, journal = {Clinical Microbiology and Infection}, doi = {10.1016/j.cmi.2018.10.020}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236866}, pages = {857-864}, year = {2019}, abstract = {Objectives Parapneumonic pleural effusions/empyema (PPE/PE) are severe complications of community-acquired pneumonia. We investigated the bacterial aetiology and incidence of paediatric PPE/PE in Germany after the introduction of universal pneumococcal conjugate vaccine (PCV) immunization for infants. Methods Children <18 years of age hospitalized with pneumonia-associated PPE/PE necessitating pleural drainage or persisting >7 days were reported to the German Surveillance Unit for Rare Diseases in Childhood between October 2010 and June 2017. All bacteria detected in blood or pleural fluid (by culture/PCR) were included, with serotyping for Streptococcus pneumoniae. Results The median age of all 1447 PPE/PE patients was 5 years (interquartile range 3-10). In 488 of the 1447 children with PPE/PE (34\%), 541 bacteria (>40 species) were detected. Aerobic gram-positive cocci accounted for 469 of 541 bacteria detected (87\%); these were most frequently Streptococcus pneumoniae (41\%), Streptococcus pyogenes (19\%) and Staphylococcus aureus (6\%). Serotype 3 accounted for 45\% of 78 serotyped S. pneumoniae strains. Annual PPE/PE incidence varied between 14 (95\%CI 12-16) and 18 (95\%CI 16-21) PPE/PE per million children. Incidence of S. pneumoniae PPE/PE decreased from 3.5 (95\%CI 2.5-4.6) per million children in 2010/11 to 1.5 (95\%CI 0.9-2.4) in 2013/14 (p 0.002), followed by a re-increase to 2.2 (95\%CI 1.5-3.2) by 2016/17 (p 0.205). Conclusions In the era of widespread PCV immunization, cases of paediatric PPE/PE were still caused mainly by S. pneumoniae and, increasingly, by S. pyogenes. The re-increase in the incidence of PPE/PE overall and in S. pneumoniae-associated PPE/PE indicates ongoing changes in the bacterial aetiology and requires further surveillance.}, language = {en} } @article{SteinKantarjianGoekbugetetal.2019, author = {Stein, Anthony S. and Kantarjian, Hagop and G{\"o}kbuget, Nicola and Bargou, Ralf and Litzow, Mark R. and Rambaldi, Alessandro and Ribera, Josep-Maria and Zhang, Alicia and Zimmerman, Zachary and Zugmaier, Gerhard and Topp, Max S.}, title = {Blinatumomab for Acute Lymphoblastic Leukemia Relapse after Allogeneic Hematopoietic Stem Cell Transplantation}, series = {Biology of Blood and Marrow Transplantation}, volume = {25}, journal = {Biology of Blood and Marrow Transplantation}, doi = {10.1016/j.bbmt.2019.04.010}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239510}, pages = {1498-1504}, year = {2019}, abstract = {Patients with relapsed/refractory (R/R) acute lymphoblastic leukemia (ALL) following allogeneic hematopoietic stem cell transplantation (alloHSCT) have a poor prognosis, and alternative therapies are needed for this patient population. Blinatumomab, a bispecific T cell engager immunotherapy, was evaluated in an open-label, single-arm, phase II study of adults with R/R Philadelphia chromosome-negative B cell precursor ALL and resulted in a rate of complete remission (CR) or CR with partial hematologic recovery of peripheral blood counts (CRh) of 43\% within 2 treatment cycles. We conducted an exploratory analysis to determine the efficacy and safety of blinatumomab in 64 patients who had relapsed following alloHSCT before enrollment in the phase II study. Forty-five percent of the patients (29 of 64) achieved a CR/CRh within the first 2 cycles of treatment, 22 of whom had a minimal residual disease (MRD) response (including 19 with a complete MRD response). After 1 year and 3 years of follow-up, the median relapse-free survival was 7.4 months for patients who achieved CR/CRh in the first 2 cycles, and the median overall survival was 8.5 months; overall survival rate (Kaplan-Meier estimate) was 36\% at 1 year and 18\% at 3 years. Grade 3 and 4 adverse events were reported in 20 patients (31\%) and 28 patients (44\%), respectively, with grade 3 and 4 neurologic events in 8 and 2 patients, respectively, and grade 3 cytokine release syndrome in 2 patients. Eight patients had fatal adverse events, including 5 due to infections. Seven patients had grade ≤ 3 graft-versus-host disease during the study, none of which resulted in the discontinuation of blinatumomab or hospitalization. Our data suggest that blinatumomab is an effective salvage therapy in this patient population.}, language = {en} } @article{HochleitnerChenBlumetal.2018, author = {Hochleitner, Gernot and Chen, Fei and Blum, Carina and Dalton, Paul D. and Amsden, Brian and Groll, J{\"u}rgen}, title = {Melt electrowriting below the critical translation speed to fabricate crimped elastomer scaffolds with non-linear extension behaviour mimicking that of ligaments and tendons}, series = {Acta Biomaterialia}, volume = {72}, journal = {Acta Biomaterialia}, doi = {10.1016/j.actbio.2018.03.023}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-320846}, pages = {110-120}, year = {2018}, abstract = {Abstract Ligaments and tendons are comprised of aligned, crimped collagen fibrils that provide tissue-specific mechanical properties with non-linear extension behaviour, exhibiting low stress at initial strain (toe region behaviour). To approximate this behaviour, we report fibrous scaffolds with sinusoidal patterns by melt electrowriting (MEW) below the critical translation speed (CTS) by exploitation of the natural flow behaviour of the polymer melt. More specifically, we synthesised photopolymerizable poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) (p(LLA-co-ε-CL-co-AC)) and poly(ε-caprolactone-co-acryloyl carbonate) (p(ε-CL-co-AC)) by ring-opening polymerization (ROP). Single fibre (f{\O} = 26.8 ± 1.9 µm) tensile testing revealed a customisable toe region with Young's Moduli ranging from E = 29 ± 17 MPa for the most crimped structures to E = 314 ± 157 MPa for straight fibres. This toe region extended to scaffolds containing multiple fibres, while the sinusoidal pattern could be influenced by printing speed. The synthesized polymers were cytocompatible and exhibited a tensile strength of σ = 26 ± 7 MPa after 104 cycles of preloading at 10\% strain while retaining the distinct toe region commonly observed in native ligaments and tendon tissue. Statement of Significance Damaged tendons and ligaments are serious and frequently occurring injuries worldwide. Recent therapies, including autologous grafts, still have severe disadvantages leading to a demand for synthetic alternatives. Materials envisioned to induce tendon and ligament regeneration should be degradable, cytocompatible and mimic the ultrastructural and mechanical properties of the native tissue. Specifically, we utilised photo-cross-linkable polymers for additive manufacturing (AM) with MEW. In this way, we were able to direct-write cytocompatible fibres of a few micrometres thickness into crimp-structured elastomer scaffolds that mimic the non-linear biomechanical behaviour of tendon and ligament tissue.}, language = {en} } @article{TesfamariamJakobWoeckeletal.2019, author = {Tesfamariam, Y. and Jakob, T. and W{\"o}ckel, A. and Adams, A. and Weigl, A. and Monsef, I. and Kuhr, K. and Skoetz, N.}, title = {Adjuvant bisphosphonates or RANK-ligand inhibitors for patients with breast cancer and bone metastases: A systematic review and network meta-analysis}, series = {Critical Reviews in Oncology / Hematology}, volume = {137}, journal = {Critical Reviews in Oncology / Hematology}, doi = {10.1016/j.critrevonc.2019.02.004}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-240827}, pages = {1-8}, year = {2019}, abstract = {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.}, language = {en} } @article{BrinkerHeklerHauschildetal.2019, author = {Brinker, Titus J. and Hekler, Achim and Hauschild, Axel and Berking, Carola and Schilling, Bastian and Enk, Alexander H. and Haferkamp, Sebastian and Karoglan, Ante and von Kalle, Christof and Weichenthal, Michael and Sattler, Elke and Schadendorf, Dirk and Gaiser, Maria R. and Klode, Joachim and Utikal, Jochen S.}, title = {Comparing artificial intelligence algorithms to 157 German dermatologists: the melanoma classification benchmark}, series = {European Journal of Cancer}, volume = {111}, journal = {European Journal of Cancer}, doi = {10.1016/j.ejca.2018.12.016}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-220569}, pages = {30-37}, year = {2019}, abstract = {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.}, language = {en} } @article{BrinkerHeklerEnketal.2019, author = {Brinker, Titus J. and Hekler, Achim and Enk, Alexander H. and Berking, Carola and Haferkamp, Sebastian and Hauschild, Axel and Weichenthal, Michael and Klode, Joachim and Schadendorf, Dirk and Holland-Letz, Tim and von Kalle, Christof and Fr{\"o}hling, Stefan and Schilling, Bastian and Utikal, Jochen S.}, title = {Deep neural networks are superior to dermatologists in melanoma image classification}, series = {European Journal of Cancer}, volume = {119}, journal = {European Journal of Cancer}, doi = {10.1016/j.ejca.2019.05.023}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-220539}, pages = {11-17}, year = {2019}, abstract = {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).}, language = {en} } @phdthesis{Zuber2024, author = {Zuber, Jonas Maximilian}, title = {Evaluation von Sedierungen und Allgemeinan{\"a}sthesien zur Durchf{\"u}hrung bildgebender Verfahren bei S{\"a}uglingen bis zum 6. Lebensmonat}, doi = {10.25972/OPUS-36111}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-361111}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Vorliegende Untersuchung am Universit{\"a}tsklinikum W{\"u}rzburg sowie die Befragung von An{\"a}sthesisten/An{\"a}sthesistinnen im Raum der 3 DACH-L{\"a}nder zeigen, dass bildgebende Verfahren bei S{\"a}uglingen mit einer niedrigen Rate an Komplikationen, zumeist in medikament{\"o}ser Sedierung mit Propofol, durchgef{\"u}hrt werden. Wie international {\"u}blich ist im S{\"a}uglingsalter die Magnetresonanztomographie das bildgebende Verfahren der Wahl und wird, mit {\"u}berzeugender H{\"a}ufigkeit, erfolgreich durchgef{\"u}hrt. Die Untersuchung am Universit{\"a}tsklinikum W{\"u}rzburg legt nahe, dass m{\"a}nnliche S{\"a}uglinge h{\"a}ufiger eine Bildgebung ben{\"o}tigen und h{\"a}ufiger h{\"o}heren ASA-Kategorie zugeschrieben werden. Dabei scheinen sie auch h{\"a}ufiger Komplikationen zu erleben und bed{\"u}rfen 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 Abl{\"a}ufe und Prozesszeiten strukturiert und optimiert werden k{\"o}nnen, da beispielsweise die Nach{\"u}berwachung entf{\"a}llt. Vorstellbar w{\"a}re beispielsweise, mehrere S{\"a}uglinge zum gleichen Zeitpunkt ins MRT zu bestellen, um gegebenenfalls den am fr{\"u}hesten eingeschlafenen S{\"a}ugling vorzuziehen. Diese Methode sollte zuk{\"u}nftig Einzug in die wissenschaftliche Untersuchung von bildgebenden Verfahren bei S{\"a}uglingen finden. Die Umfrage im deutschsprachigen Raum zeigt eine Leitlinien-gerechte Betreuung von S{\"a}uglingen f{\"u}r bildgebende Verfahren, die mit einer hohen Qualit{\"a}t, und zumeist erfolgreich von erfahrenen An{\"a}sthesisten/An{\"a}sthesistinnen durchgef{\"u}hrt wird. Eventuelle Verbesserungen k{\"o}nnen im Bereich der Ausbildung nachfolgender {\"A}rztinnen/{\"A}rzte und in der h{\"a}ufigeren Verwendung der „feed-and-sleep" Methode liegen, die vielen Kollegen/Kolleginnen bekannt ist, aber nur selten durchgef{\"u}hrt wird. Ziel ist eine qualitativ hochwertige, schnellstm{\"o}glich durchgef{\"u}hrte Bildgebung, die ohne oder mit der niedrigst m{\"o}glichen Dosierung eines sedierenden Medikamentes zu erreichen ist.}, subject = {Sedierung}, language = {de} } @phdthesis{Dalkmann2024, author = {Dalkmann, Theresa}, title = {Evaluierung prognostischer und pr{\"a}diktiver Biomarker beim neoadjuvant vorbehandelten Rektumkarzinom}, doi = {10.25972/OPUS-36336}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-363368}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Fragestellung. Osteopontin (OPN) kann im Blut nachgewiesen werden und wird bei vielen Tumorentit{\"a}ten 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 pr{\"a}diktive 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 {\"U}berleben (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 Pr{\"a}diktivit{\"a}t OPNs f{\"u}r 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 {\"U}berleben. F{\"u}r c-Met und HGF ergab sich keine Korrelation zu initialen klinischen Daten und Therapieansprechen. Die logistische Regression ergab keinen pr{\"a}diktiven Wert. Schlussfolgerung. Die Plasmakonzentration von OPN besitzt prognostische und pr{\"a}diktive Wertigkeit beim LARC. Die Konzentrationen von c-Met und HGF sind nicht prognostisch f{\"u}r das {\"U}berleben oder pr{\"a}diktiv f{\"u}r das Therapieansprechen.}, subject = {Biomarker}, language = {de} } @phdthesis{Junghanns2024, author = {Junghanns, Lara Madeleine}, title = {Resistenzmechanismen gegen Amphotericin B in humanpathogenen Hefepilzen}, doi = {10.25972/OPUS-36986}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369861}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die 2009 erstmals entdeckte Spezies C. auris erlangte binnen k{\"u}rzester Zeit zunehmend weltweite Aufmerksamkeit. Vor allem die Tendenz der Multiresistenzentwicklung und das rasche Ausl{\"o}sen 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 {\"u}ber den Wirkmechanismus von Amphotericin B in Hefepilzen zu erlangen. Anhand der zun{\"a}chst durchgef{\"u}hrten 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 St{\"a}mmen eine Mutation im ERG4 Gen an Position 576, welche nicht eindeutig als alleinige Ursache f{\"u}r die verminderte Amphotericin B-Empfindlichkeit festgelegt werden konnte. Dennoch wurde im Rahmen eines Survival Assays bei beiden Amphotericin B-resistenten Isolaten anf{\"a}nglich eine konzentrationsabh{\"a}ngige Aktivit{\"a}t gegen{\"u}ber Amphotericin B festgestellt, bevor ein Nachwachsen der Kulturen beobachtet wurde. Somit wurde die Vermutung aufgestellt, dass lediglich ein Teil der aufgebrachten Candida-Zellen abget{\"o}tet wird und dies in einer Vermehrung der {\"u}berlebenden 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 Wirkungsverst{\"a}rkung des Polyens hervorgerufen wird. Diese Sensitivit{\"a}tssteigerung ist allgemein bei allen C. auris Isolaten zu beobachten, f{\"a}llt bei resistenten St{\"a}mmen jedoch deutlich st{\"a}rker aus. Hierdurch kam die Annahme auf, dass Amphotericin B-Resistenzen auch in m{\"o}glichen Ver{\"a}nderungen des Sphingolipid-Haushaltes begr{\"u}ndet sein k{\"o}nnten. Dar{\"u}ber hinaus scheint Myriocin keinen Einfluss auf Fluconazol-resistente oder FKS-mutierte Echinocandin-resistente C. auris St{\"a}mme zu haben. Das ebenfalls untersuchte und von Myriocin abgeleitete Medikament Fingolimod hatte jedoch ebenfalls keinen wirkungsverst{\"a}rkenden Effekt. Allerdings reagierte ein Großteil der C. auris Isolate (57,6 \%) sensitiv gegen{\"u}ber 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 m{\"o}glich}, subject = {Candida}, language = {de} } @article{JeanclosAlbersenRamosetal.2019, author = {Jeanclos, Elisabeth and Albersen, Monique and Ramos, R{\´u}ben J. J. and Raab, Annette and Wilhelm, Christian and Hommers, Leif and Lesch, Klaus-Peter and Verhoeven-Duif, Nanda M. and Gohla, Antje}, title = {Improved cognition, mild anxiety-like behavior and decreased motor performance in pyridoxal phosphatase-deficient mice}, series = {BBA - Molecular Basis of Disease}, volume = {1865}, journal = {BBA - Molecular Basis of Disease}, doi = {10.1016/j.bbadis.2018.08.018}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-323396}, pages = {193-205}, year = {2019}, abstract = {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.}, language = {en} } @article{MuellerCosentinoFoerstneretal.2018, author = {M{\"u}ller, Laura S. M. and Cosentino, Ra{\´u}l O. and F{\"o}rstner, Konrad U. and Guizetti, Julien and Wedel, Carolin and Kaplan, Noam and Janzen, Christian J. and Arampatzi, Panagiota and Vogel, J{\"o}rg and Steinbiss, Sascha and Otto, Thomas D. and Saliba, Antoine-Emmanuel and Sebra, Robert P. and Siegel, T. Nicolai}, title = {Genome organization and DNA accessibility control antigenic variation in trypanosomes}, series = {Nature}, volume = {563}, journal = {Nature}, doi = {10.1038/s41586-018-0619-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224265}, pages = {121-125}, year = {2018}, abstract = {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.}, language = {en} } @article{MunzRichterLoosetal.2018, author = {Munz, Matthias and Richter, Gesa M. and Loos, Bruno G. and Jepsen, S{\o}ren and Divaris, Kimon and Offenbacher, Steven and Teumer, Alexander and Holtfreter, Birte and Kocher, Thomas and Bruckmann, Corinna and Jockel-Schneider, Yvonne and Graetz, Christian and Munoz, Loreto and Bhandari, Anita and Tennstedt, Stephanie and Staufenbiel, Ingmar and van der Velde, Nathalie and Uitterlinden, Andr{\´e} G. and de Groot, Lisette C. P. G. M. and Wellmann, J{\"u}rgen and Berger, Klaus and Krone, Bastian and Hoffmann, Per and Laudes, Matthias and Lieb, Wolfgang and Andre, Franke and Dommisch, Henrik and Erdmann, Jeanette and Schaefer, Arne S.}, title = {Genome-wide association meta-analysis of coronary artery disease and periodontitis reveals a novel shared risk locus}, series = {Scientific Reports}, volume = {8}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-31980-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-231647}, year = {2018}, abstract = {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.}, language = {en} } @phdthesis{PaetzelgebDitter2024, author = {P{\"a}tzel [geb. Ditter], Katharina Sabine}, title = {Molekulare Charakterisierung eines Mitgliedes der TNF-Rezeptor-Superfamilie des Fuchsbandwurmes \(Echinococcus\) \(multilocularis\)}, doi = {10.25972/OPUS-36939}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369397}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die alveol{\"a}re Echinokokkose (AE), die durch den Fuchsbandwurm Echinococcus multilocularis verursacht wird, ist eine seltene jedoch schwere und oft t{\"o}dlich verlaufende Erkrankung. Aufgrund der sp{\"a}ten Diagnosestellung sind kurative Behandlungsmethoden h{\"a}ufig nicht durchf{\"u}hrbar und als einzige Behandlungsm{\"o}glichkeit bleibt eine lebenslange und nebenwirkungsreiche Therapie mit Benzimidazolen. Verbesserte Therapieoptionen durch die Entwicklung neuer Medikamente sind dringend notwendig. Hierf{\"u}r 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 Entschl{\"u}sselung des Echinococcus-Genoms gab Hinweise darauf, dass ein Mitglied der Tumornekrosefaktor-Rezeptor-Superfamilie, jedoch kein endogener TNF α {\"a}hnlicher Ligand im Genom kodiert wird. Ein Mitglied der TNFR-Superfamilie des Fuchsbandwurmes (EmTNFR) wurde in dieser Arbeit als membranst{\"a}ndiger Rezeptor mit einer intrazellul{\"a}ren Todesdom{\"a}ne (DD) und hoher {\"A}hnlichkeit 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 intrazellul{\"a}re Todesdom{\"a}ne besitzt. emtnfr-v3 verwendet einen alternativen Transkriptionstart und wird von den letzten 3 Exons von emtnfr kodiert. emtnfr-v3, kodiert ein Protein ohne extrazellul{\"a}re Region, aber mit intrazellul{\"a}rer Todesdom{\"a}ne. Ein l{\"o}slicher 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 urspr{\"u}nglichen Form der TNF-Rezeptoren entspricht. Mitglieder eines intrazellul{\"a}ren 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 Prim{\"a}rzellen und im Metazestoden (MZ), dem pathogenen Stadium f{\"u}r 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. Zus{\"a}tzlich wurde in whole-mount in situ Hybridisierungs-Versuchen eine ubiquit{\"a}re Expression von emtnfr in der Germinalschicht des MZ sowie eine Spezifit{\"a}t von emtnfr f{\"u}r den MZ, welcher urs{\"a}chlich f{\"u}r die AE ist, nachgewiesen. Somit scheinen sowohl EmTNFR als auch TNF-α eine wichtige Funktion bei der Entwicklung und Etablierung des Fuchsbandwurmes w{\"a}hrend der fr{\"u}hen Phase der Infektion des Zwischenwirtes zu haben. TNF-α k{\"o}nnte ein weiterer Faktor f{\"u}r den ausgepr{\"a}gten 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 {\"u}ber die Bindung von Wirts-TNF-α bei der fr{\"u}hen Entwicklung des Echincoccus-Metazestoden eine Rolle spielt.}, subject = {Fuchsbandwurm}, language = {de} } @phdthesis{Reissland2024, author = {Reissland, Michaela}, title = {USP10 is a \(de\) \(novo\) tumour-specific regulator of β-Catenin and contributes to cancer stem cell maintenance and tumour progression}, doi = {10.25972/OPUS-31957}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-319579}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {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.}, subject = {Biomedizin}, language = {en} } @article{MorimotoShimadaSugimotoOtowaetal.2018, author = {Morimoto, Yoshiro and Shimada-Sugimoto, Mihoko and Otowa, Takeshi and Yoshida, Shintaro and Kinoshita, Akira and Mishima, Hiroyuki and Yamaguchi, Naohiro and Mori, Takatoshi and Imamura, Akira and Ozawa, Hiroki and Kurotaki, Naohiro and Ziegler, Christiane and Domschke, Katharina and Deckert, J{\"u}rgen and Umekage, Tadashi and Tochigi, Mamoru and Kaiya, Hisanobu and Okazaki, Yuji and Tokunaga, Katsushi and Sasaki, Tsukasa and Yoshiura, Koh-ichiro and Ono, Shinji}, title = {Whole-exome sequencing and gene-based rare variant association tests suggest that PLA2G4E might be a risk gene for panic disorder}, series = {Translational Psychiatry}, volume = {8}, journal = {Translational Psychiatry}, doi = {10.1038/s41398-017-0088-0}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224192}, year = {2018}, abstract = {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.}, language = {en} } @article{NerreterLetschertGoetzetal.2019, author = {Nerreter, Thomas and Letschert, Sebastian and G{\"o}tz, Ralph and Doose, S{\"o}ren and Danhof, Sophia and Einsele, Hermann and Sauer, Markus and Hudecek, Michael}, title = {Super-resolution microscopy reveals ultra-low CD19 expression on myeloma cells that triggers elimination by CD19 CAR-T}, series = {Nature Communications}, volume = {10}, journal = {Nature Communications}, doi = {10.1038/s41467-019-10948-w}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232258}, year = {2019}, abstract = {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.}, language = {en} } @article{OdinChaudhuriVolkmannetal.2018, author = {Odin, Per and Chaudhuri, K. Ray and Volkmann, Jens and Antonini, Angelo and Storch, Alexander and Dietrichs, Espen and Pirtošek, Zvezdan and Henriksen, Tove and Horne, Malcolm and Devos, David and Bergquist, Filip}, title = {Viewpoint and practical recommendations from a movement disorder specialist panel on objective measurement in the clinical management of Parkinson's disease}, series = {npj Parkinson's Disease}, volume = {4}, journal = {npj Parkinson's Disease}, doi = {10.1038/s41531-018-0051-7}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-234435}, year = {2018}, abstract = {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.}, language = {en} } @article{ScholzCosgareaSuesskindetal.2018, author = {Scholz, S. L. and Cosgarea, I. and S{\"u}ßkind, D. and Murali, R. and M{\"o}ller, I. and Reis, H. and Leonardelli, S. and Schilling, B. and Schimming, T. and Hadaschik, E. and Franklin, C. and Paschen, A. and Sucker, A. and Steuhl, K. P. and Schadendorf, D. and Westekemper, H. and Griewank, K. G.}, title = {NF1 mutations in conjunctival melanoma}, series = {British Journal of Cancer}, volume = {118}, journal = {British Journal of Cancer}, doi = {10.1038/s41416-018-0046-5}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233329}, pages = {1243-1247}, year = {2018}, abstract = {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}, language = {en} } @article{BruchhagenJarickMewisetal.2018, author = {Bruchhagen, Christin and Jarick, Marcel and Mewis, Carolin and Hertlein, Tobias and Niemann, Silke and Ohlsen, Knut and Peters, Georg and Planz, Oliver and Ludwig, Stephan and Ehrhardt, Christina}, title = {Metabolic conversion of CI-1040 turns a cellular MEK-inhibitor into an antibacterial compound}, series = {Scientific Reports}, volume = {8}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-27445-7}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-221648}, year = {2018}, abstract = {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.}, language = {en} } @article{CastilhoHochleitnerWilsonetal.2018, author = {Castilho, Miguel and Hochleitner, Gernot and Wilson, Wouter and van Rietbergen, Bert and Dalton, Paul D. and Groll, J{\"u}rgen and Malda, Jos and Ito, Keita}, title = {Mechanical behavior of a soft hydrogel reinforced with three-dimensional printed microfibre scaffolds}, series = {Scientific Reports}, volume = {8}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-19502-y}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-222280}, year = {2018}, abstract = {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.}, language = {en} } @article{AlZabenMedyukhinaDietrichetal.2019, author = {Al-Zaben, Naim and Medyukhina, Anna and Dietrich, Stefanie and Marolda, Alessandra and H{\"u}nniger, Kerstin and Kurzai, Oliver and Figge, Marc Thilo}, title = {Automated tracking of label-free cells with enhanced recognition of whole tracks}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-39725-x}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-221093}, year = {2019}, abstract = {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.}, language = {en} } @article{deJongDinizSalomaetal.2018, author = {de Jong, Simone and Diniz, Mateus Jose Abdalla and Saloma, Andiara and Gadelha, Ary and Santoro, Marcos L. and Ota, Vanessa K. and Noto, Cristiano and Curtis, Charles and Newhouse, Stephen J. and Patel, Hamel and Hall, Lynsey S. and O'Reilly, Paul F. and Belangero, Sintia I. and Bressan, Rodrigo A. and Breen, Gerome}, title = {Applying polygenic risk scoring for psychiatric disorders to a large family with bipolar disorder and major depressive disorder}, series = {Communications Biology}, volume = {1}, journal = {Communications Biology}, organization = {Major Depressive Disorder and Bipolar Disorder Working Groups of the Psychiatric Genomics Consortium}, doi = {10.1038/s42003-018-0155-y}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223622}, year = {2018}, abstract = {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.}, language = {en} } @article{DekkerDiekstraPulitetal.2019, author = {Dekker, Annelot M. and Diekstra, Frank P. and Pulit, Sara L. and Tazelaar, Gijs H. P. and van der Spek, Rick A. and van Rheenen, Wouter and van Eijk, Kristel R. and Calvo, Andrea and Brunetti, Maura and Van Damme, Philip and Robberecht, Wim and Hardiman, Orla and McLaughlin, Russell and Chi{\`o}, Adriano and Sendtner, Michael and Ludolph, Albert C. and Weishaupt, Jochen H. and Pardina, Jesus S. Mora and van den Berg, Leonard H. and Veldink, Jan H.}, title = {Exome array analysis of rare and low frequency variants in amyotrophic lateral sclerosis}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-42091-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223686}, year = {2019}, abstract = {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.}, language = {en} } @article{DiehlSchmidLicataGoldhardtetal.2019, author = {Diehl-Schmid, Janine and Licata, Abigail and Goldhardt, Oliver and F{\"o}rstl, Hans and Yakushew, Igor and Otto, Markus and Anderl-Straub, Sarah and Beer, Ambros and Ludolph, Albert Christian and Landwehrmeyer, Georg Bernhard and Levin, Johannes and Danek, Adrian and Fliessbach, Klaus and Spottke, Annika and Fassbender, Klaus and Lyros, Epameinondas and Prudlo, Johannes and Krause, Bernd Joachim and Volk, Alexander and Edbauer, Dieter and Schroeter, Matthias Leopold and Drzezga, Alexander and Kornhuber, Johannes and Lauer, Martin and Grimmer, Timo}, title = {FDG-PET underscores the key role of the thalamus in frontotemporal lobar degeneration caused by C9ORF72 mutations}, series = {Translational Psychiatry}, volume = {9}, journal = {Translational Psychiatry}, organization = {FTLDc Study Group}, doi = {10.1038/s41398-019-0381-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-225308}, year = {2019}, abstract = {C9ORF72 mutations are the most common cause of familial frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). MRI studies have investigated structural changes in C9ORF72-associated FTLD (C9FTLD) and provided first insights about a prominent involvement of the thalamus and the cerebellum. Our multicenter, 18F-fluorodeoxyglucose positron-emission tomography study of 22 mutation carriers with FTLD, 22 matched non-carriers with FTLD, and 23 cognitively healthy controls provided valuable insights into functional changes in C9FTLD: compared to non-carriers, mutation carriers showed a significant reduction of glucose metabolism in both thalami, underscoring the key role of the thalamus in C9FTLD. Thalamic metabolism did not correlate with disease severity, duration of disease, or the presence of psychotic symptoms. Against our expectations we could not demonstrate a cerebellar hypometabolism in carriers or non-carriers. Future imaging and neuropathological studies in large patient cohorts are required to further elucidate the central role of the thalamus in C9FTLD.}, language = {en} } @article{DietrichKrugKrastletal.2019, author = {Dietrich, Thomas and Krug, Ralf and Krastl, Gabriel and Tomson, Philip L.}, title = {Restoring the unrestorable! Developing coronal tooth tissue with a minimally invasive surgical extrusion technique}, series = {British Dental Journal}, volume = {226}, journal = {British Dental Journal}, doi = {10.1038/s41415-019-0268-9}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-225333}, pages = {789-793}, year = {2019}, abstract = {Surgical extrusion is a recognised treatment option for teeth that have insufficient coronal tooth structure remaining due to deep caries, resorption or traumatic injury. However, the technique has not been widely adopted, arguably because extraction of a severely compromised tooth may be difficult to achieve in a gentle and predictable way. In this paper, we present our novel approach to surgical extrusion and subsequent management of teeth using a vertical extraction system (Benex), which has become the method of choice in the authors' practice for many teeth that would otherwise be deemed unrestorable. We describe the clinical procedure in detail and discuss the advantages and disadvantages compared to alternative approaches, including surgical crown lengthening and orthodontic extrusion.}, language = {en} } @article{HauerPoppSchoelleretal.2018, author = {Hauer, Nadine N. and Popp, Bernt and Schoeller, Eva and Schuhmann, Sarah and Heath, Karen E. and Hisado-Oliva, Alfonso and Klinger, Patricia and Kraus, Cornelia and Trautmann, Udo and Zenker, Martin and Zweier, Christiane and Wiesener, Antje and Jamra, Rami Abou and Kunstmann, Erdmute and Wieczorek, Dagmar and Uebe, Steffen and Ferrazzi, Fulvia and B{\"u}ttner, Christian and Ekici, Arif B. and Rauch, Anita and Sticht, Heinrich and D{\"o}rr, Helmuth-G{\"u}nther and Reis, Andr{\´e} and Thiel, Christian T.}, title = {Clinical relevance of systematic phenotyping and exome sequencing in patients with short stature}, series = {Genetics in Medicine}, volume = {20}, journal = {Genetics in Medicine}, doi = {10.1038/gim.2017.159}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-227888}, pages = {630-638}, year = {2018}, abstract = {Purpose Short stature is a common condition of great concern to patients and their families. Mostly genetic in origin, the underlying cause often remains elusive due to clinical and genetic heterogeneity. Methods We systematically phenotyped 565 patients where common nongenetic causes of short stature were excluded, selected 200 representative patients for whole-exome sequencing, and analyzed the identified variants for pathogenicity and the affected genes regarding their functional relevance for growth. Results By standard targeted diagnostic and phenotype assessment, we identified a known disease cause in only 13.6\% of the 565 patients. Whole-exome sequencing in 200 patients identified additional mutations in known short-stature genes in 16.5\% of these patients who manifested only part of the symptomatology. In 15.5\% of the 200 patients our findings were of significant clinical relevance. Heterozygous carriers of recessive skeletal dysplasia alleles represented 3.5\% of the cases. Conclusion A combined approach of systematic phenotyping, targeted genetic testing, and whole-exome sequencing allows the identification of the underlying cause of short stature in at least 33\% of cases, enabling physicians to improve diagnosis, treatment, and genetic counseling. Exome sequencing significantly increases the diagnostic yield and consequently care in patients with short stature.}, language = {en} } @article{LudwigDelforgeFaconetal.2018, author = {Ludwig, Heinz and Delforge, Michel and Facon, Thierry and Einsele, Hermann and Gay, Francesca and Moreau, Philippe and Avet-Loiseau, Herv{\´e} and Boccadoro, Mario and Hajek, Roman and Mohty, Mohamad and Cavo, Michele and Dimopoulos, Meletios A and San-Miguel, Jes{\´u}s F and Terpos, Evangelos and Zweegman, Sonja and Garderet, Laurent and Mateos, Mar{\´i}a-Victoria and Cook, Gordon and Leleu, Xavier and Goldschmidt, Hartmut and Jackson, Graham and Kaiser, Martin and Weisel, Katja and van de Donk, Niels W. C. J. and Waage, Anders and Beksac, Meral and Mellqvist, Ulf H. and Engelhardt, Monika and Caers, Jo and Driessen, Christoph and Blad{\´e}, Joan and Sonneveld, Pieter}, title = {Prevention and management of adverse events of novel agents in multiple myeloma: a consensus of the European Myeloma Network}, series = {Leukemia}, volume = {32}, journal = {Leukemia}, doi = {10.1038/s41375-018-0040-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-237338}, pages = {1542-1560}, year = {2018}, abstract = {During the last few years, several new drugs have been introduced for treatment of patients with multiple myeloma, which have significantly improved the treatment outcome. All of these novel substances differ at least in part in their mode of action from similar drugs of the same drug class, or are representatives of new drug classes, and as such present with very specific side effect profiles. In this review, we summarize these adverse events, provide information on their prevention, and give practical guidance for monitoring of patients and for management of adverse events.}, language = {en} } @phdthesis{Machwart2024, author = {Machwart, Khaled}, title = {Modulatorischer Einfluss von Levosimendan bei dem Isch{\"a}mie-Reperfusionsschaden auf die myokardiale Mitochondrienfunktion}, doi = {10.25972/OPUS-36102}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-361021}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die vorliegende Studie untersuchte den Effekt von Levosimendan auf die mitochondriale Funktionen im Herzmuskel, insbesondere im Zusammenhang mit dem Isch{\"a}mie/Reperfusions-Schaden. Methoden: In der Studie wurde ein retrogrades Langendorff-Modell verwendet, um die Auswirkungen von Levosimendan, dem Isch{\"a}mie/Reperfusions-Schaden sowie deren Kombination auf die mitochondrialen Funktionen im Herzmuskel zu untersuchen. Dazu wurden vier verschiedene Gruppen von Rattenherzen entsprechend den experimentellen Bedingungen perfundiert, und ihre Funktionen wurden analysiert. Ergebnisse: Der Isch{\"a}mie/Reperfusions-Schaden beeintr{\"a}chtigte die myokardiale Ventrikelfunktion. Zus{\"a}tzlich wurde eine Hypopolarisation des mithochondrialen Membranpotentials in den mit Levosimendan oder Isch{\"a}mie behandelten Gruppen festgestellt. Die ATP-Synthese in den Gruppen mit Levosimendan und Isch{\"a}mie war reduziert. Schlussfolgerung: Levosimendan zeigt signifikante Einfl{\"u}sse auf die Atmungsfunktion der mitochondrialen Komplexe IV und V sowie auf das Membranpotential. Diese Ph{\"a}nomene k{\"o}nnten einem mito-K+ ATP-abh{\"a}ngigen Mechanismus zugrunde liegen. Obwohl Levosimendan w{\"a}hrend des Isch{\"a}mie/Reperfusionsschadens eine protektive Wirkung hinsichtlich einer Ca2+- {\"U}berlastung aufweist, bleibt der kumulative Einfluss der beeintr{\"a}chtigten ATP-Generierung auf die gesamte Myokardfunktion zu kl{\"a}ren.}, subject = {Isch{\"a}mie}, language = {de} } @article{PrustyGulveGovindetal.2018, author = {Prusty, Bhupesh K. and Gulve, Nitish and Govind, Sheila and Krueger, Gerhard R. F. and Feichtinger, Julia and Larcombe, Lee and Aspinall, Richard and Ablashi, Dharam V. and Toro, Carla T.}, title = {Active HHV-6 Infection of Cerebellar Purkinje Cells in Mood Disorders}, series = {Frontiers in Microbiology}, volume = {9}, journal = {Frontiers in Microbiology}, doi = {10.3389/fmicb.2018.01955}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369222}, year = {2018}, abstract = {Early-life infections and associated neuroinflammation is incriminated in the pathogenesis of various mood disorders. Infection with human roseoloviruses, HHV-6A and HHV-6B, allows viral latency in the central nervous system and other tissues, which can later be activated causing cognitive and behavioral disturbances. Hence, this study was designed to evaluate possible association of HHV-6A and HHV-6B activation with three different groups of psychiatric patients. DNA qPCR, immunofluorescence and FISH studies were carried out in post-mortem posterior cerebellum from 50 cases each of bipolar disorder (BPD), schizophrenia, 15 major depressive disorder (MDD) and 50 appropriate control samples obtained from two well-known brain collections (Stanley Medical Research Institute). HHV-6A and HHV-6B late proteins (indicating active infection) and viral DNA were detected more frequently (p < 0.001 for each virus) in human cerebellum in MDD and BPD relative to controls. These roseolovirus proteins and DNA were found less frequently in schizophrenia cases. Active HHV-6A and HHV-6B infection in cerebellar Purkinje cells were detected frequently in BPD and MDD cases. Furthermore, we found a significant association of HHV-6A infection with reduced Purkinje cell size, suggesting virus-mediated abnormal Purkinje cell function in these disorders. Finally, gene expression analysis of cerebellar tissue revealed changes in pathways reflecting an inflammatory response possibly to HHV-6A infection. Our results provide molecular evidence to support a role for active HHV-6A and HHV-6B infection in BPD and MDD.}, language = {en} } @article{TichaMoosWajantetal.2018, author = {Ticha, Olga and Moos, Lukas and Wajant, Harald and Bekeredjian-Ding, Isabelle}, title = {Expression of Tumor Necrosis Factor Receptor 2 Characterizes TLR9-Driven Formation of Interleukin-10-Producing B Cells}, series = {Frontiers in Immunology}, volume = {8}, journal = {Frontiers in Immunology}, doi = {10.3389/fimmu.2017.01951}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-241323}, year = {2018}, abstract = {B cell-derived interleukin-10 (IL-10) production has been described as a hallmark for regulatory function in B lymphocytes. However, there is an ongoing debate on the origin of IL-10-secreting B cells and lack of specific surface markers has turned into an important obstacle for studying human B regulatory cells. In this study, we propose that tumor necrosis factor receptor 2 (TNFR2) expression can be used for enrichment of IL-10-secreting B cells. Our data confirm that IL-10 production can be induced by TLR9 stimulation with CpG ODN and that IL-10 secretion accompanies differentiation of peripheral blood B cells into plasma blasts. We further show that CpG ODN stimulation induces TNFR2 expression, which correlates with IL-10 secretion and terminal differentiation. Indeed, flow cytometric sorting of TNFR2+ B cells revealed that TNFR2+ and TNFR2- fractions correspond to IL-10+ and IL-10- fractions, respectively. Furthermore, CpG-induced TNFR2+ B cells were predominantly found in the IgM+ CD27+ B cell subset and spontaneously released immunoglobulin. Finally, our data corroborate the functional impact of TNFR2 by demonstrating that stimulation with a TNFR2 agonist significantly augments IL-10 and IL-6 production in B cells. Altogether, our data highlight a new role for TNFR2 in IL-10-secreting human B lymphocytes along with the potential to exploit this finding for sorting and isolation of this currently ill-defined B cell subset.}, language = {en} } @article{KlotzHigginsSchaubmaretal.2019, author = {Klotz, Peter and Higgins, Paul G. and Schaubmar, Andreas R. and Failing, Klaus and Leidner, Ursula and Seifert, Harald and Scheufen, Sandra and Semmler, Torsten and Ewers, Christa}, title = {Seasonal Occurrence and Carbapenem Susceptibility of Bovine Acinetobacter baumannii in Germany}, series = {Frontiers in Microbiology}, volume = {10}, journal = {Frontiers in Microbiology}, doi = {10.3389/fmicb.2019.00272}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-325927}, year = {2019}, abstract = {Acinetobacter baumannii is one of the leading causes of nosocomial infections in humans. To investigate its prevalence, distribution of sequence types (STs), and antimicrobial resistance in cattle, we sampled 422 cattle, including 280 dairy cows, 59 beef cattle, and 83 calves over a 14-month period. Metadata, such as the previous use of antimicrobial agents and feeding, were collected to identify putative determining factors. Bacterial isolates were identified via MALDI-TOF/MS and PCR, antimicrobial susceptibility was evaluated via VITEK2 and antibiotic gradient tests, resistance genes were identified by PCR. Overall, 15.6\% of the cattle harbored A. baumannii, predominantly in the nose (60.3\% of the A. baumannii isolates). It was more frequent in dairy cows (21.1\%) than in beef cattle (6.8\%) and calves (2.4\%). A seasonal occurrence was shown with a peak between May and August. The rate of occurrence of A. baumannii was correlated with a history of use of 3rd generation cephalosporins in the last 6 months prior to sampling Multilocus sequence typing (Pasteur scheme) revealed 83 STs among 126 unique isolates. Nine of the bovine STs have previously been implicated in human infections. Besides known intrinsic resistance of the species, the isolates did not show additional resistance to the antimicrobial substances tested, including carbapenems. Our data suggest that cattle are not a reservoir for nosocomial A. baumannii but carry a highly diverse population of this species. Nevertheless, some STs seem to be able to colonize both cattle and humans.}, language = {en} } @article{SchroeterPawelkeBiseniusetal.2018, author = {Schroeter, Matthias L. and Pawelke, Sarah and Bisenius, Sandrine and Kynast, Jana and Schuemberg, Katharina and Polyakova, Maryna and Anderl-Straub, Sarah and Danek, Adrian and Fassbender, Klaus and Jahn, Holger and Jessen, Frank and Kornhuber, Johannes and Lauer, Martin and Prudlo, Johannes and Schneider, Anja and Uttner, Ingo and Th{\"o}ne-Otto, Angelika and Otto, Markus and Diehl-Schmid, Janine}, title = {A Modified Reading the Mind in the Eyes Test Predicts Behavioral Variant Frontotemporal Dementia Better Than Executive Function Tests}, series = {Frontiers in Aging Neuroscience}, volume = {10}, journal = {Frontiers in Aging Neuroscience}, organization = {FTLD Study Group Germany}, doi = {10.3389/fnagi.2018.00011}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-234254}, year = {2018}, abstract = {Behavioral variant frontotemporal dementia (bvFTD) is characterized by deep alterations in behavior and personality. Although revised diagnostic criteria agree for executive dysfunction as most characteristic, impairments in social cognition are also suggested. The study aimed at identifying those neuropsychological and behavioral parameters best discriminating between bvFTD and healthy controls. Eighty six patients were diagnosed with possible or probable bvFTD according to Rascovsky et al. (2011) and compared with 43 healthy age-matched controls. Neuropsychological performance was assessed with a modified Reading the Mind in the Eyes Test (RMET), Stroop task, Trail Making Test (TMT), Hamasch-Five-Point Test (H5PT), and semantic and phonemic verbal fluency tasks. Behavior was assessed with the Apathy Evaluation Scale, Frontal Systems Behavioral Scale, and Bayer Activities of Daily Living Scale. Each test's discriminatory power was investigated by Receiver Operating Characteristic curves calculating the area under the curve (AUC). bvFTD patients performed significantly worse than healthy controls in all neuropsychological tests. Discriminatory power (AUC) was highest in behavioral questionnaires, high in verbal fluency tasks and the RMET, and lower in executive function tests such as the Stroop task, TMT and H5PT. As fluency tasks depend on several cognitive functions, not only executive functions, results suggest that the RMET discriminated better between bvFTD and control subjects than other executive tests. Social cognition should be incorporated into diagnostic criteria for bvFTD in the future, such as in the International Classification of Diseases (ICD)-11, as already suggested in the Diagnostic and Statistical Manual for Mental Disorders (DSM)-5.}, language = {en} } @article{LangFuellsackWajant2018, author = {Lang, Isabell and F{\"u}llsack, Simone and Wajant, Harald}, title = {Lack of Evidence for a Direct Interaction of Progranulin and Tumor Necrosis Factor Receptor-1 and Tumor Necrosis Factor Receptor-2 From Cellular Binding Studies}, series = {Frontiers in Immunology}, volume = {9}, journal = {Frontiers in Immunology}, doi = {10.3389/fimmu.2018.00793}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236373}, year = {2018}, abstract = {Progranulin (PGRN) is a secreted anti-inflammatory protein which can be processed by neutrophil proteases to various granulins. It has been reported that at least a significant portion of the anti-inflammatory effects of PGRN is due to direct high affinity binding to tumor necrosis factor receptor-1 (TNFR1) and TNFR2 and inhibition of tumor necrosis factor (TNF)-induced TNFR1/2 signaling. Two studies failed to reproduce the interaction of TNFR1 and TNFR2 with PGRN, but follow up reports speculated that this was due to varying experimental circumstances and/or the use of PGRN from different sources. However, even under consideration of these speculations, there is still a striking discrepancy in the literature between the concentrations of PGRN needed to inhibit TNF signaling and the concentrations required to block TNF binding to TNFR1 and TNFR2. While signaling events induced by 0.2-2 nM of TNF have been efficiently inhibited by low, near to equimolar concentrations (0.5-2.5 nM) of PGRN in various studies, the reported inhibitory effects of PGRN on TNF-binding to TNFR1/2 required a huge excess of PGRN (100-1,000-fold). Therefore, we investigated the effect of PGRN on TNF binding to TNFR1 and TNFR2 in highly sensitive cellular binding studies. Unlabeled TNF inhibited >95\% of the specific binding of a Gaussia princeps luciferase (GpL) fusion protein of TNF to TNFR1 and TNFR2 and blocked binding of soluble GpL fusion proteins of TNFR1 and TNFR2 to membrane TNF expressing cells to >95\%, too. Purified PGRN, however, showed in both assays no effect on TNF-TNFR1/2 interaction even when applied in huge excess. To rule out that tags and purification- or storage-related effects compromise the potential ability of PGRN to bind TNF receptors, we directly co-expressed PGRN, and as control TNF, in TNFR1- and TNFR2-expressing cells and looked for binding of GpL-TNF. While expression of TNF strongly inhibited binding of GpL-TNF to TNFR1/2, co-expression of PGRN had not effect on the ability of the TNFR1/2-expressing cells to bind TNF.}, language = {en} } @article{WeissZieglerFliesseretal.2018, author = {Weiss, Esther and Ziegler, Sabrina and Fliesser, Mirjam and Schmitt, Anna-Lena and H{\"u}nniger, Kerstin and Kurzai, Oliver and Morton, Charles-Oliver and Einsele, Hermann and Loeffler, Juergen}, title = {First Insights in NK—DC Cross-Talk and the Importance of Soluble Factors During Infection With Aspergillus fumigatus}, series = {Frontiers in Cellular and Infection Microbiology}, volume = {8}, journal = {Frontiers in Cellular and Infection Microbiology}, doi = {10.3389/fcimb.2018.00288}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233565}, year = {2018}, abstract = {Invasive aspergillosis (IA) is an infectious disease caused by the fungal pathogen Aspergillus fumigatus that mainly affects immunocompromised hosts. To investigate immune cell cross-talk during infection with A. fumigatus, we co-cultured natural killer (NK) cells and dendritic cells (DC) after stimulation with whole fungal structures, components of the fungal cell wall, fungal lysate or ligands for distinct fungal receptors. Both cell types showed activation after stimulation with fungal components and were able to transfer activation signals to the counterpart not stimulated cell type. Interestingly, DCs recognized a broader spectrum of fungal components and thereby initiated NK cell activation when those did not recognize fungal structures. These experiments highlighted the supportive function of DCs in NK cell activation. Furthermore, we focused on soluble DC mediated NK cell activation and showed that DCs stimulated with the TLR2/Dectin-1 ligand zymosan could maximally stimulate the expression of CD69 on NK cells. Thus, we investigated the influence of both receptors for zymosan, Dectin-1 and TLR2, which are highly expressed on DCs but show only minimal expression on NK cells. Specific focus was laid on the question whether Dectin-1 or TLR2 signaling in DCs is important for the secretion of soluble factors leading to NK cell activation. Our results show that Dectin-1 and TLR2 are negligible for NK cell activation. We conclude that besides Dectin-1 and TLR2 other receptors on DCs are able to compensate for the missing signal.}, language = {en} } @article{SchurigHaeuslerGrittneretal.2019, author = {Schurig, Johannes and Haeusler, Karl Georg and Grittner, Ulrike and Nolte, Christian H. and Fiebach, Jochen B. and Audebert, Heinrich J. and Endres, Matthias and Rocco, Andrea}, title = {Frequency of Hemorrhage on Follow Up Imaging in Stroke Patients Treated With rt-PA Depending on Clinical Course}, series = {Frontiers in Neurology}, volume = {10}, journal = {Frontiers in Neurology}, doi = {10.3389/fneur.2019.00368}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-234947}, year = {2019}, abstract = {Background: According to current guidelines, stroke patients treated with rt-PA should undergo brain imaging to exclude intracerebral bleeding 24 h after thrombolysis, before the start of medical secondary prevention. However, the usefulness of routine follow-up imaging with regard to changes in therapeutic management in patients without neurological deterioration is unclear. We hypothesized that follow up brain imaging solely to exclude bleeding in patients who clinically improved after rt-PA application may not be necessary. Methods: Retrospective single-center analysis including stroke patients treated with rt-PA. Records were reviewed for hemorrhagic transformation one day after systemic thrombolysis and brain imaging-based changes in therapeutic management. Twenty-four hour after thrombolysis patients were divided into four groups: (1) increased NIHSS score; (2) unchanged NIHSS score; (3) improved NIHSS score and; (4) NIHSS score = 0. Results: Out of 188 patients (mean age 73 years, 100 female) receiving rt-PA, 32 (17\%) had imaging-proven hemorrhagic transformation including 11 (6\%) patients with parenchymal hemorrhage. Patients in group (1, 2) more often had hypertension (p = 0.015) and more often had parenchymal hemorrhage (9 vs. 4\%; p < 0.206) compared to group (3, 4) and imaging-based changes in therapeutic management were more frequent (19\% vs. 6\%; p = 0.007). Patients of group (3, 4) had no changes in therapeutic management in 94\% of the cases. Patients in group (4) had no hemorrhagic transformation in routine follow-up brain imaging. Conclusions: Frequency of hemorrhagic transformation in Routine follow-up brain imaging and consecutive changes in therapeutic management were different depending on clinical course measured by NHISS score.}, language = {en} } @phdthesis{CruzdeCasas2024, author = {Cruz de Casas, Paulina}, title = {Sphingolipids as modulators of T cell function}, doi = {10.25972/OPUS-35969}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-359698}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {The immune system is responsible for the preservation of homeostasis whenever a given organism is exposed to distinct kinds of perturbations. Given the complexity of certain organisms like mammals, and the diverse types of challenges that they encounter (e.g. infection or disease), the immune system evolved to harbor a great variety of distinct immune cell populations with specialized functions. For instance, the family of T cells is sub-divided into conventional (Tconv) and unconventional T cells (UTCs). Tconv form part of the adaptive arm of the immune system and are comprised of αβ CD4+ or CD8+ cells that differentiate from na{\"i}ve to effector and memory populations upon activation and are essential during infection and cancer. Furthermore, UTCs, which include γδ T cells, NKT and MAIT, are involved in innate and adaptive immune responses, due to their dual mode of activation, through cytokines (innate-like) or TCR (adaptive), and function. Despite our understanding of the basic functions of T cells in several contexts, a great number of open questions related to their basic biology remain. For instance, the mechanism behind the differentiation of na{\"i}ve CD4+ and CD8+ T cells into effector and memory populations is not fully understood. Moreover, the exact function and relevance of distinct UTC subpopulations in a physiological context have not been fully clarified. Here, we investigated the factors mediating na{\"i}ve CD8+ T cell differentiation into effector and memory cells. By using flow cytometry, mass spectrometry, enzymatic assays, and transgenic mouse models, we found that the membrane bound enzyme sphingomyelin-phosphodiesterase acid-like 3b (Smpdl3b) is crucial for the maintenance of memory CD8+ T cells. Our data show that the absence of Smpdl3b leads to diminished CD8+ T cell memory, and a loss of stem-like memory populations due to an aggravated contraction. Our scRNA-seq data suggest that Smpdl3b could be involved in clathrinmediated endocytosis through modulation of Huntingtin interacting protein 1 (Hip1) levels, likely regulating TCR-independent signaling events. Furthermore, in this study we explored the role of UTCs in lymph node-specific immune responses. By using transgenic mouse models for photolabeling, lymph node transplantation models, infection models and flow cytometry, we demonstrate that S1P regulates the migration of tissue-derived UTC from tissues to draining lymph nodes, resulting in heterogeneous immune responses mounted by lymph nodes draining different tissues. Moreover, our unbiased scRNAseq and single lineage-deficient mouse models analysis revealed that all UTC lineages (γδ T cells, NKT and MAIT) are organized in functional units, based on transcriptional homogeneity, shared microanatomical location and migratory behavior, and numerical and functional redundancy. Taken together, our studies describe additional cell intrinsic (Smpdl3b) and extrinsic (S1Pmediated migration) functions of sphingolipid metabolism modulating T cell biology. We propose the S1P/S1PR1/5 signaling axis as the potential survival pathway for Smpdl3b+ memory CD8+ T cells and UTCs, mainly in lymph nodes. Possibly, Smpdl3b regulates S1P/S1PR signaling by balancing ligandreceptor endocytosis, while UTCs migrate to lymph nodes during homeostasis to be exposed to specific levels of S1P that assure their maintenance. Our results are clinically relevant, since several drugs modulating the S1P/S1PR signaling axis or the levels of Smpdl3b are currently used to treat human diseases, such as multiple sclerosis and B cell-mediated diseases. We hope that our discoveries will inspire future studies focusing on sphingolipid metabolism in immune cell biology.}, subject = {T-Lymphozyt}, language = {en} } @phdthesis{Wussmann2024, author = {Wußmann, Maximiliane}, title = {Humane organotypische 3D Modelle des Malignen Melanoms als in vitro Testsystem f{\"u}r die Bewertung der Wirksamkeit von anti-Tumor Therapeutika}, doi = {10.25972/OPUS-36100}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-361005}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Das maligne Melanom, eine der seltensten, aber gleichzeitig auch die t{\"o}dlichste dermatologische Malignit{\"a}t, gekennzeichnet durch die Neigung zu einer fr{\"u}hen Metastasierung sowie die rasche Entwicklung von Therapieresistenzen, z{\"a}hlt zu den Tumorentit{\"a}ten mit dem h{\"o}chsten Anstieg der Inzidenz weltweit. Mausmodelle werden h{\"a}ufig verwendet, um die Melanomagenese zu erforschen und neue effektive therapeutische Strategien zu entwickeln, spiegeln die menschliche Physiologie allerdings nur unzureichend wider. In zweidimensionalen (2D) Zellkulturen mangelt es dagegen an wichtigen Komponenten der Mikroumgebung des Tumors und dem dreidimensionalen Gewebekontext. Um dieses Manko zu beheben und die Entwicklung von auf den Menschen {\"u}bertragbaren Tumormodellen in der onkologischen Forschung voranzutreiben, wurde als Alternative zu Zellkulturen und Tierversuchen humane organotypische dreidimensionale (3D) Melanom-Modelle als in vitro Testsystem f{\"u}r die Bewertung der Wirksamkeit von anti-Tumor Therapeutika entwickelt. Im Zuge dieser Arbeit konnte das in vitro Melanom-Modell entscheidend weiterentwickelt werden. So konnten Modelle unterschiedlichster Komplexit{\"a}t etabliert werden, wobei abh{\"a}ngig von der Fragestellung einfachere epidermale bis hin zu unterschiedlich komplexen Vollhautmodellen Anwendung finden. Durch Simulation der Tumor-Mikroumgebung eignen sich diese zur pr{\"a}klinischen Validierung neuer Tumor-Therapeutika, sowie der Erforschung pathologischer Vorg{\"a}nge, von der Tumor-Formierung bis zur Metastasierung. Zudem konnten erfolgreich unterschiedlichste humane Melanomzelllinien ins Modell integriert werden; dadurch, dass sich diese durch ihre Treibermutationen, die zur Krankheitsentstehung beitragen, unterscheiden, stellen sie unterschiedliche Anspr{\"u}che an potentielle therapeutische Angriffspunkte und erm{\"o}glichen das Widerspiegeln vieler Melanom-Subtypen im Modell. Ferner ist es m{\"o}glich, verschiedene Stadien der Tumor-Entwicklung {\"u}ber die Zugabe von Melanomzellen in Einzelsuspension bzw. von Melanom-Sph{\"a}roiden widerzuspiegeln. Es konnte f{\"u}r bestimmte Therapie-Ans{\"a}tze, wie zielgerichtete Therapien, z.B. die Gabe von sich in der Klinik im Einsatz befindlicher BRAF-/MEK-Inhibitoren, gezeigt werden, dass sich die etablierten Modelle hervorragend als pr{\"a}klinische Testsysteme zur Wirksamkeitsbewertung eignen. Zudem bieten sich einzigartige M{\"o}glichkeiten, um die Interaktion humaner Tumorzellen und gesunder Zellen in einem Gewebeverband zu untersuchen. Ferner konnten drei neue technische Analyse-Verfahren zur nicht-invasiven Detektion der Tumor- Pro- und Regression, Beurteilung der Wirksamkeit von potenziellen Anti-Tumor-Therapien sowie der Evaluierung des Tumor-Metabolismusses implementiert werden. Perspektivisch erm{\"o}glichen immun-kompetente Melanom-Modelle die Austestung neuer Immun- und Zelltherapien in einem voll humanen System; gleichzeitig leisten die etablierten Modelle einen signifikanten Beitrag zur Reduktion von Tierexperimenten.}, subject = {Melanom}, language = {de} } @phdthesis{Adhikari2024, author = {Adhikari, Bikash}, title = {Targeted degradation of Myc-interacting oncoproteins}, doi = {10.25972/OPUS-31732}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-317326}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {The hallmark oncoprotein Myc is a major driver of tumorigenesis in various human cancer entities. However, Myc's structural features make it challenging to develop small molecules against it. A promising strategy to indirectly inhibit the function of Myc is by targeting its interactors. Many Myc-interacting proteins have reported scaffolding functions which are difficult to target using conventional occupancy- driven inhibitors. Thus, in this thesis, the proteolysis targeting chimera (PROTAC) approach was used to target two oncoproteins interacting with Myc which promote the oncogenicity of Myc, Aurora-A and WDR5. PROTACs are bifunctional small molecules that bind to the target protein with one ligand and recruit a cellular E3- ligase with the other ligand to induce target degradation via the ubiquitin- proteasome system. So far, the most widely used E3-ligases for PROTAC development are Cereblon (CRBN) and von Hippel-Lindau tumor suppressor (VHL). Furthermore, there are cases of incompatibility between some E3-ligases and proteins to bring about degradation. Hence there is a need to explore new E3- ligases and a demand for a tool to predict degradative E3-ligases for the target protein in the PROTAC field. In the first part, a highly specific mitotic kinase Aurora-A degrader, JB170, was developed. This compound utilized Aurora-A inhibitor alisertib as the target ligand and thalidomide as the E3-ligase CRBN harness. The specificity of JB170 and the ternary complex formation was supported by the interactions between Aurora-A and CRBN. The PROTAC-mediated degradation of Aurora-A induced a distinct S- phase defect rather than mitotic arrest, shown by its catalytic inhibition. The finding demonstrates that Aurora-A has a non-catalytic role in the S-phase. Furthermore, the degradation of Aurora-A led to apoptosis in various cancer cell lines. In the second part, two different series of WDR5 PROTACs based on two protein- protein inhibitors of WDR5 were evaluated. The most efficient degraders from both series recruited VHL as a E3-ligase and showed partial degradation of WDR5. In addition, the degradation efficiency of the PROTACs was significantly affected by the linker nature and length, highlighting the importance of linker length and composition in PROTAC design. The degraders showed modest proliferation defects at best in cancer cell lines. However, overexpression of VHL increased the degradation efficiency and the antiproliferative effect of the PROTACs. In the last part, a rapamycin-based assay was developed to predict the degradative E3-ligase for a target. The assay was validated using the WDR5/VHL and Aurora- A/CRBN pairs. The result that WDR5 is degraded by VHL but not CRBN and Aurora-A is degraded by CRBN, matches observations made with PROTACs. This technique will be used in the future to find effective tissue-specific and essential E3-ligases for targeted degradation of oncoproteins using PROTACs. Collectively, the work presented here provides a strategy to improve PROTAC development and a starting point for developing Aurora-A and WDR5 PROTACs for cancer therapy.}, subject = {Degradation}, language = {en} } @phdthesis{Bakirci2024, author = {Bakirci, Ezgi}, title = {Development of \(In\) \(vitro\) Models for Tissue Engineering Applications Using a High-Resolution 3D Printing Technology}, doi = {10.25972/OPUS-25164}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-251645}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {In vitro models mimic the tissue-specific anatomy and play essential roles in personalized medicine and disease treatments. As a sophisticated manufacturing technology, 3D printing overcomes the limitations of traditional technologies and provides an excellent potential for developing in vitro models to mimic native tissue. This thesis aims to investigate the potential of a high-resolution 3D printing technology, melt electrowriting (MEW), for fabricating in vitro models. MEW has a distinct capacity for depositing micron size fibers with a defined design. In this thesis, three approaches were used, including 1) extending the MEW polymer library for different biomedical applications, 2) developing in vitro models for evaluation of cell growth and migration toward the different matrices, and 3) studying the effect of scaffold designs and biochemical cues of microenvironments on cells. First, we introduce the MEW processability of (AB)n and (ABAC)n segmented copolymers, which have thermally reversible network formulation based on physical crosslinks. Bisurea segments are combined with hydrophobic poly(dimethylsiloxane) (PDMS) or hydrophilic poly(propylene oxide)-poly(ethylene oxide)-poly(propylene oxide) (PPO-PEG-PPO) segments to form the (AB)n segmented copolymers. (ABAC)n segmented copolymers contain all three segments: in addition to bisurea, both hydrophobic and hydrophilic segments are available in the same polymer chain, resulting in tunable mechanical and biological behaviors. MEW copolymers either support cells attachment or dissolve without cytotoxic side effects when in contact with the polymers at lower concentrations, indicating that this copolymer class has potential in biological applications. The unique biological and surface properties, transparency, adjustable hydrophilicity of these copolymers could be beneficial in several in vitro models. The second manuscript addresses the design and development of a melt electrowritten competitive 3D radial migration device. The approach differs from most of the previous literature, as MEW is not used here to produce cell invasive scaffolds but to fabricate an in vitro device. The device is utilized to systematically determine the matrix which promotes cell migration and growth of glioblastoma cells. The glioblastoma cell migration is tested on four different Matrigel concentrations using a melt electrowritten radial device. The glioblastoma U87 cell growth and migration increase at Matrigel concentrations 6 and 8 mg mL-1 In the development of this radial device, the accuracy, and precision of melt electrowritten circular shapes were investigated. The results show that the printing speed and design diameter are essential parameters for the accuracy of printed constructs. It is the first instance where MEW is used for the production of in vitro devices. The influence of biochemical cues and scaffold designs on astrocytes and glioblastoma is investigated in the last manuscript. A fiber comprising the box and triangle-shaped pores within MEW scaffolds are modified with biochemical cues, including RGD and IKVAV peptides using a reactive NCO-sP(EO-stat-PO) macromer. The results show that astrocytes and glioblastoma cells exhibit different phenotypes on scaffold designs and peptide-coated scaffolds.}, subject = {3D-Druck}, language = {en} } @article{SnaebjornssonSchulze2018, author = {Snaebjornsson, Marteinn T and Schulze, Almut}, title = {Non-canonical functions of enzymes facilitate cross-talk between cell metabolic and regulatory pathways}, series = {Experimental \& Molecular Medicine}, volume = {50}, journal = {Experimental \& Molecular Medicine}, doi = {10.1038/s12276-018-0065-6}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-238763}, pages = {1-16}, year = {2018}, abstract = {The metabolic rewiring that occurs during cell transformation is a hallmark of cancer. It is diverse in different cancers as it reflects different combinations of oncogenic drivers, tumor suppressors, and the microenvironment. Metabolic rewiring is essential to cancer as it enables uncontrolled proliferation and adaptation to the fluctuating availability of nutrients and oxygen caused by poor access to the vasculature due to tumor growth and a foreign microenvironment encountered during metastasis. Increasing evidence now indicates that the metabolic state in cancer cells also plays a causal role in tumor growth and metastasis, for example through the action of oncometabolites, which modulate cell signaling and epigenetic pathways to promote malignancy. In addition to altering the metabolic state in cancer cells, some multifunctional enzymes possess non-metabolic functions that also contribute to cell transformation. Some multifunctional enzymes that are highly expressed in cancer, such as pyruvate kinase M2 (PKM2), have non-canonical functions that are co-opted by oncogenic signaling to drive proliferation and inhibit apoptosis. Other multifunctional enzymes that are frequently downregulated in cancer, such as fructose-bisphosphatase 1 (FBP1), are tumor suppressors, directly opposing mitogenic signaling via their non-canonical functions. In some cases, the enzymatic and non-canonical roles of these enzymes are functionally linked, making the modulation of non-metabolic cellular processes dependent on the metabolic state of the cell.}, language = {en} } @article{SirtlKnollDieuThuyetal.2018, author = {Sirtl, Simon and Knoll, Gertrud and Dieu Thuy, Trinh and Lang, Isabell and Siegmund, Daniela and Gross, Stefanie and Schuler-Thurner, Beatrice and Neubert, Patrick and Jantsch, Jonathan and Wajant, Harald and Ehrenschwender, Martin}, title = {Hypertonicity-enforced BCL-2 addiction unleashes the cytotoxic potential of death receptors}, series = {Oncogene}, volume = {37}, journal = {Oncogene}, doi = {10.1038/s41388-018-0265-5}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-238327}, pages = {4122-4136}, year = {2018}, abstract = {Attempts to exploit the cytotoxic activity of death receptors (DR) for treating cancer have thus far been disappointing. DR activation in most malignant cells fails to trigger cell death and may even promote tumor growth by activating cell death-independent DR-associated signaling pathways. Overcoming apoptosis resistance is consequently a prerequisite for successful clinical exploitation of DR stimulation. Here we show that hyperosmotic stress in the tumor microenvironment unleashes the deadly potential of DRs by enforcing BCL-2 addiction of cancer cells. Hypertonicity robustly enhanced cytotoxicity of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and other DR ligands in various cancer entities. Initial events in TRAIL DR signaling remained unaffected, but hypertonic conditions unlocked activation of the mitochondrial death pathway and thus amplified the apoptotic signal. Mechanistically, we demonstrate that hyperosmotic stress imposed a BCL-2-addiction on cancer cells to safeguard the integrity of the outer mitochondrial membrane (OMM), essentially exhausting the protective capacity of BCL-2-like pro-survival proteins. Deprivation of these mitochondrial safeguards licensed DR-generated truncated BH3-interacting domain death agonist (tBID) to activate BCL-2-associated X protein (BAX) and initiated mitochondrial outer membrane permeabilization (MOMP). Our work highlights that hyperosmotic stress in the tumor environment primes mitochondria for death and lowers the threshold for DR-induced apoptosis. Beyond TRAIL-based therapies, our findings could help to strengthen the efficacy of other apoptosis-inducing cancer treatment regimens.}, language = {en} } @article{StegmannReichertsAndreattaetal.2019, author = {Stegmann, Yannik and Reicherts, Philipp and Andreatta, Marta and Pauli, Paul and Wieser, Matthias J.}, title = {The effect of trait anxiety on attentional mechanisms in combined context and cue conditioning and extinction learning}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-45239-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239394}, year = {2019}, abstract = {Sensory processing and attention allocation are shaped by threat, but the role of trait-anxiety in sensory processing as a function of threat predictability remains incompletely understood. Therefore, we measured steady-state visual evoked potentials (ssVEPs) as an index of sensory processing of predictable and unpredictable threat cues in 29 low (LA) and 29 high (HA) trait-anxious participants during a modified NPU-paradigm followed by an extinction phase. Three different contextual cues indicated safety (N), predictable (P) or unpredictable threat (U), while foreground cues signalled shocks in the P-condition only. All participants allocated increased attentional resources to the central P-threat cue, replicating previous findings. Importantly, LA individuals exhibited larger ssVEP amplitudes to contextual threat (U and P) than to contextual safety cues, while HA individuals did not differentiate among contextual cues in general. Further, HA exhibited higher aversive ratings of all contexts compared to LA. These results suggest that high trait-anxious individuals might be worse at discriminating contextual threat stimuli and accordingly overestimate the probability and aversiveness of unpredictable threat. These findings support the notion of aberrant sensory processing of unpredictable threat in anxiety disorders, as this processing pattern is already evident in individuals at risk of these disorders.}, language = {en} } @article{SolimandoBrandlMattenheimeretal.2018, author = {Solimando, A G and Brandl, A and Mattenheimer, K and Graf, C and Ritz, M and Ruckdeschel, A and St{\"u}hmer, T and Mokhtari, Z and Rudelius, M and Dotterweich, J and Bittrich, M and Desantis, V and Ebert, R and Trerotoli, P and Frassanito, M A and Rosenwald, A and Vacca, A and Einsele, H and Jakob, F and Beilhack, A}, title = {JAM-A as a prognostic factor and new therapeutic target in multiple myeloma}, series = {Leukemia}, volume = {32}, journal = {Leukemia}, doi = {10.1038/leu.2017.287}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239069}, pages = {736-743}, year = {2018}, abstract = {Cell adhesion in the multiple myeloma (MM) microenvironment has been recognized as a major mechanism of MM cell survival and the development of drug resistance. Here we addressed the hypothesis that the protein junctional adhesion molecule-A (JAM-A) may represent a novel target and a clinical biomarker in MM. We evaluated JAM-A expression in MM cell lines and in 147 MM patient bone marrow aspirates and biopsies at different disease stages. Elevated JAM-A levels in patient-derived plasma cells were correlated with poor prognosis. Moreover, circulating soluble JAM-A (sJAM-A) levels were significantly increased in MM patients as compared with controls. Notably, in vitro JAM-A inhibition impaired MM migration, colony formation, chemotaxis, proliferation and viability. In vivo treatment with an anti-JAM-A monoclonal antibody (αJAM-A moAb) impaired tumor progression in a murine xenograft MM model. These results demonstrate that therapeutic targeting of JAM-A has the potential to prevent MM progression, and lead us to propose JAM-A as a biomarker in MM, and sJAM-A as a serum-based marker for clinical stratification.}, language = {en} } @article{SiegmundEhrenschwenderWajant2018, author = {Siegmund, Daniela and Ehrenschwender, Martin and Wajant, Harald}, title = {TNFR2 unlocks a RIPK1 kinase activity-dependent mode of proinflammatory TNFR1 signaling}, series = {Cell Death \& Disease}, volume = {9}, journal = {Cell Death \& Disease}, doi = {10.1038/s41419-018-0973-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-238034}, year = {2018}, abstract = {TNF is not only a major effector molecule of PAMP/DAMP-activated macrophages, but also regulates macrophage function and viability. We recently demonstrated that TNFR2 triggers necroptosis in macrophages with compromised caspase activity by two cooperating mechanisms: induction of endogenous TNF with subsequent stimulation of TNFR1 and depletion of cytosolic TRAF2-cIAP complexes. Here we show that TNFR2 activation in caspase-inhibited macrophages results in the production of endogenous TNF and TNFR1 stimulation followed by upregulation of A20, TRAF1, IL-6, and IL-1β. Surprisingly, TNFR1-mediated induction of IL-6 and IL-1β was clearly evident in response to TNFR2 stimulation but occurred not or only weakly in macrophages selectively and directly stimulated via TNFR1. Moreover, TNFR2-induced TNFR1-mediated gene induction was largely inhibited by necrostatin-1, whereas upregulation of A20 and TRAF1 by direct and exclusive stimulation of TNFR1 remained unaffected by this compound. Thus, treatment with TNFR2/ZVAD enables TNFR1 in macrophages to stimulate gene induction via a pathway requiring RIPK1 kinase activity. TNFR2/ZVAD-induced production of IL-6 and IL-1β was largely blocked in necroptosis-resistant MLKL- and RIPK3-deficient macrophages, whereas induction of A20 and TRAF1 remained unaffected. In sum, our results show that in caspase-inhibited macrophages TNFR2 not only triggers TNF/TNFR1-mediated necroptosis but also TNF/TNFR1-mediated RIPK3/MLKL-dependent and -independent gene induction.}, language = {en} } @article{VaethWangEcksteinetal.2019, author = {Vaeth, Martin and Wang, Yin-Hu and Eckstein, Miriam and Yang, Jun and Silverman, Gregg J. and Lacruz, Rodrigo S. and Kannan, Kasthuri and Feske, Stefan}, title = {Tissue resident and follicular Treg cell differentiation is regulated by CRAC channels}, series = {Nature Communications}, volume = {10}, journal = {Nature Communications}, doi = {10.1038/s41467-019-08959-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232148}, year = {2019}, abstract = {T regulatory (Treg) cells maintain immunological tolerance and organ homeostasis. Activated Treg cells differentiate into effector Treg subsets that acquire tissue-specific functions. Ca2+ influx via Ca2+ release-activated Ca2+ (CRAC) channels formed by STIM and ORAI proteins is required for the thymic development of Treg cells, but its function in mature Treg cells remains unclear. Here we show that deletion of Stim1 and Stim2 genes in mature Treg cells abolishes Ca2+ signaling and prevents their differentiation into follicular Treg and tissue-resident Treg cells. Transcriptional profiling of STIM1/STIM2-deficient Treg cells reveals that Ca2+ signaling regulates transcription factors and signaling pathways that control the identity and effector differentiation of Treg cells. In the absence of STIM1/STIM2 in Treg cells, mice develop a broad spectrum of autoantibodies and fatal multiorgan inflammation. Our findings establish a critical role of CRAC channels in controlling lineage identity and effector functions of Treg cells.}, language = {en} } @article{SulzerCassidyHorgaetal.2018, author = {Sulzer, David and Cassidy, Clifford and Horga, Guillermo and Kang, Un Jung and Fahn, Stanley and Casella, Luigi and Pezzoli, Gianni and Langley, Jason and Hu, Xiaoping P. and Zucca, Fabio A. and Isaias, Ioannis U. and Zecca, Luigi}, title = {Neuromelanin detection by magnetic resonance imaging (MRI) and its promise as a biomarker for Parkinson's disease}, series = {npj Parkinson's Disease}, volume = {4}, journal = {npj Parkinson's Disease}, doi = {10.1038/s41531-018-0047-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-240207}, year = {2018}, abstract = {The diagnosis of Parkinson's disease (PD) occurs after pathogenesis is advanced and many substantia nigra (SN) dopamine neurons have already died. Now that therapies to block this neuronal loss are under development, it is imperative that the disease be diagnosed at earlier stages and that the response to therapies is monitored. Recent studies suggest this can be accomplished by magnetic resonance imaging (MRI) detection of neuromelanin (NM), the characteristic pigment of SN dopaminergic, and locus coeruleus (LC) noradrenergic neurons. NM is an autophagic product synthesized via oxidation of catecholamines and subsequent reactions, and in the SN and LC it increases linearly during normal aging. In PD, however, the pigment is lost when SN and LC neurons die. As shown nearly 25 years ago by Zecca and colleagues, NM's avid binding of iron provides a paramagnetic source to enable electron and nuclear magnetic resonance detection, and thus a means for safe and noninvasive measure in living human brain. Recent technical improvements now provide a means for MRI to differentiate between PD patients and age-matched healthy controls, and should be able to identify changes in SN NM with age in individuals. We discuss how MRI detects NM and how this approach might be improved. We suggest that MRI of NM can be used to confirm PD diagnosis and monitor disease progression. We recommend that for subjects at risk for PD, and perhaps generally for older people, that MRI sequences performed at regular intervals can provide a pre-clinical means to detect presymptomatic PD.}, language = {en} } @article{UllrichWeberPostetal.2018, author = {Ullrich, M and Weber, M and Post, A M and Popp, S and Grein, J and Zechner, M and Gonz{\´a}lez, H Guerrero and Kreis, A and Schmitt, A G and {\"U}ҫeyler, N and Lesch, K-P and Schuh, K}, title = {OCD-like behavior is caused by dysfunction of thalamo-amygdala circuits and upregulated TrkB/ERK-MAPK signaling as a result of SPRED2 deficiency}, series = {Molecular Psychiatry}, volume = {23}, journal = {Molecular Psychiatry}, doi = {10.1038/mp.2016.232}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232096}, pages = {444-458}, year = {2018}, abstract = {Obsessive-compulsive disorder (OCD) is a common neuropsychiatric disease affecting about 2\% of the general population. It is characterized by persistent intrusive thoughts and repetitive ritualized behaviors. While gene variations, malfunction of cortico-striato-thalamo-cortical (CSTC) circuits, and dysregulated synaptic transmission have been implicated in the pathogenesis of OCD, the underlying mechanisms remain largely unknown. Here we show that OCD-like behavior in mice is caused by deficiency of SPRED2, a protein expressed in various brain regions and a potent inhibitor of Ras/ERK-MAPK signaling. Excessive self-grooming, reflecting OCD-like behavior in rodents, resulted in facial skin lesions in SPRED2 knockout (KO) mice. This was alleviated by treatment with the selective serotonin reuptake inhibitor fluoxetine. In addition to the previously suggested involvement of cortico-striatal circuits, electrophysiological measurements revealed altered transmission at thalamo-amygdala synapses and morphological differences in lateral amygdala neurons of SPRED2 KO mice. Changes in synaptic function were accompanied by dysregulated expression of various pre- and postsynaptic proteins in the amygdala. This was a result of altered gene transcription and triggered upstream by upregulated tropomyosin receptor kinase B (TrkB)/ERK-MAPK signaling in the amygdala of SPRED2 KO mice. Pathway overactivation was mediated by increased activity of TrkB, Ras, and ERK as a specific result of SPRED2 deficiency and not elicited by elevated brain-derived neurotrophic factor levels. Using the MEK inhibitor selumetinib, we suppressed TrkB/ERK-MAPK pathway activity in vivo and reduced OCD-like grooming in SPRED2 KO mice. Altogether, this study identifies SPRED2 as a promising new regulator, TrkB/ERK-MAPK signaling as a novel mediating mechanism, and thalamo-amygdala synapses as critical circuitry involved in the pathogenesis of OCD.}, language = {en} } @article{TrautzFrankeBohnertetal.2019, author = {Trautz, Florian and Franke, Heike and Bohnert, Simone and Hammer, Niels and M{\"u}ller, Wolf and Stassart, Ruth and Tse, Rexson and Zwirner, Johann and Dreßler, Jan and Ondruschka, Benjamin}, title = {Survival-time dependent increase in neuronal IL-6 and astroglial GFAP expression in fatally injured human brain tissue}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-48145-w}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-229037}, year = {2019}, abstract = {Knowledge on trauma survival time prior to death following a lethal traumatic brain injury (TBI) may be essential for legal purposes. Immunohistochemistry studies might allow to narrow down this survival interval. The biomarkers interleukin-6 (IL-6) and glial fibrillary acidic protein (GFAP) are well known in the clinical setting for their usability in TBI prediction. Here, both proteins were chosen in forensics to determine whether neuronal or glial expression in various brain regions may be associated with the cause of death and the survival time prior to death following TBI. IL-6 positive neurons, glial cells and GFAP positive astrocytes all concordantly increase with longer trauma survival time, with statistically significant changes being evident from three days post-TBI (p < 0.05) in the pericontusional zone, irrespective of its definite cortical localization. IL-6 staining in neurons increases significantly in the cerebellum after trauma, whereas increasing GFAP positivity is also detected in the cortex contralateral to the focal lesion. These systematic chronological changes in biomarkers of pericontusional neurons and glial cells allow for an estimation of trauma survival time. Higher numbers of IL-6 and GFAP-stained cells above threshold values in the pericontusional zone substantiate the existence of fatal traumatic changes in the brain with reasonable certainty.}, language = {en} } @article{TylekSchillingSchlegelmilchetal.2019, author = {Tylek, Tina and Schilling, Tatjana and Schlegelmilch, Katrin and Ries, Maximilian and Rudert, Maximilian and Jakob, Franz and Groll, J{\"u}rgen}, title = {Platelet lysate outperforms FCS and human serum for co-culture of primary human macrophages and hMSCs}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-40190-9}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-229174}, year = {2019}, abstract = {In vitro co-cultures of different primary human cell types are pivotal for the testing and evaluation of biomaterials under conditions that are closer to the human in vivo situation. Especially co-cultures of macrophages and mesenchymal stem cells (MSCs) are of interest, as they are both present and involved in tissue regeneration and inflammatory reactions and play crucial roles in the immediate inflammatory reactions and the onset of regenerative processes, thus reflecting the decisive early phase of biomaterial contact with the host. A co-culture system of these cell types might thus allow for the assessment of the biocompatibility of biomaterials. The establishment of such a co-culture is challenging due to the different in vitro cell culture conditions. For human macrophages, medium is usually supplemented with human serum (hS), whereas hMSC culture is mostly performed using fetal calf serum (FCS), and these conditions are disadvantageous for the respective other cell type. We demonstrate that human platelet lysate (hPL) can replace hS in macrophage cultivation and appears to be the best option for co-cultivation of human macrophages with hMSCs. In contrast to FCS and hS, hPL maintained the phenotype of both cell types, comparable to that of their respective standard culture serum, as well as the percentage of each cell population. Moreover, the expression profile and phagocytosis activity of macrophages was similar to hS.}, language = {en} } @article{StraubFreudenbergSchleicheretal.2018, author = {Straub, Tobias and Freudenberg, Marina A. and Schleicher, Ulrike and Bogdan, Christian and Gasteiger, Georg and Pircher, Hanspeter}, title = {Bacterial coinfection restrains antiviral CD8 T-cell response via LPS-induced inhibitory NK cells}, series = {Nature Communications}, volume = {9}, journal = {Nature Communications}, doi = {10.1038/s41467-018-06609-z}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-240075}, year = {2018}, abstract = {Infection of specific pathogen-free mice with lymphocytic choriomeningitis virus (LCMV) is a widely used model to study antiviral T-cell immunity. Infections in the real world, however, are often accompanied by coinfections with unrelated pathogens. Here we show that in mice, systemic coinfection with E. coli suppresses the LCMV-specific cytotoxic T-lymphocyte (CTL) response and virus elimination in a NK cell- and TLR2/4-dependent manner. Soluble TLR4 ligand LPS also induces NK cell-mediated negative CTL regulation during LCMV infection. NK cells in LPS-treated mice suppress clonal expansion of LCMV-specific CTLs by a NKG2D- or NCR1-independent but perforin-dependent mechanism. These results suggest a TLR4-mediated immunoregulatory role of NK cells during viral-bacterial coinfections.}, language = {en} } @article{WentSudSpeedyetal.2018, author = {Went, Molly and Sud, Amit and Speedy, Helen and Sunter, Nicola J. and F{\"o}rsti, Asta and Law, Philip J. and Johnson, David C. and Mirabella, Fabio and Holroyd, Amy and Li, Ni and Orlando, Giulia and Weinhold, Niels and van Duin, Mark and Chen, Bowang and Mitchell, Jonathan S. and Mansouri, Larry and Juliusson, Gunnar and Smedby, Karin E and Jayne, Sandrine and Majid, Aneela and Dearden, Claire and Allsup, David J. and Bailey, James R. and Pratt, Guy and Pepper, Chris and Fegan, Chris and Rosenquist, Richard and Kuiper, Rowan and Stephens, Owen W. and Bertsch, Uta and Broderick, Peter and Einsele, Hermann and Gregory, Walter M. and Hillengass, Jens and Hoffmann, Per and Jackson, Graham H. and J{\"o}ckel, Karl-Heinz and Nickel, Jolanta and N{\"o}then, Markus M. and da Silva Filho, Miguel Inacio and Thomsen, Hauke and Walker, Brian A. and Broyl, Annemiek and Davies, Faith E. and Hansson, Markus and Goldschmidt, Hartmut and Dyer, Martin J. S. and Kaiser, Martin and Sonneveld, Pieter and Morgan, Gareth J. and Hemminki, Kari and Nilsson, Bj{\"o}rn and Catovsky, Daniel and Allan, James M. and Houlston, Richard S.}, title = {Genetic correlation between multiple myeloma and chronic lymphocytic leukaemia provides evidence for shared aetiology}, series = {Blood Cancer Journal}, volume = {9}, journal = {Blood Cancer Journal}, doi = {10.1038/s41408-018-0162-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233627}, year = {2018}, abstract = {The clustering of different types of B-cell malignancies in families raises the possibility of shared aetiology. To examine this, we performed cross-trait linkage disequilibrium (LD)-score regression of multiple myeloma (MM) and chronic lymphocytic leukaemia (CLL) genome-wide association study (GWAS) data sets, totalling 11,734 cases and 29,468 controls. A significant genetic correlation between these two B-cell malignancies was shown (Rg = 0.4, P = 0.0046). Furthermore, four of the 45 known CLL risk loci were shown to associate with MM risk and five of the 23 known MM risk loci associate with CLL risk. By integrating eQTL, Hi-C and ChIP-seq data, we show that these pleiotropic risk loci are enriched for B-cell regulatory elements and implicate B-cell developmental genes. These data identify shared biological pathways influencing the development of CLL and, MM and further our understanding of the aetiological basis of these B-cell malignancies.}, language = {en} } @article{WenFeilWoltersetal.2018, author = {Wen, Lai and Feil, Susanne and Wolters, Markus and Thunemann, Martin and Regler, Frank and Schmidt, Kjestine and Friebe, Andreas and Olbrich, Marcus and Langer, Harald and Gawaz, Meinrad and de Wit, Cor and Feil, Robert}, title = {A shear-dependent NO-cGMP-cGKI cascade in platelets acts as an auto-regulatory brake of thrombosis}, series = {Nature Communications}, volume = {9}, journal = {Nature Communications}, doi = {10.1038/s41467-018-06638-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233616}, year = {2018}, abstract = {Mechanisms that limit thrombosis are poorly defined. One of the few known endogenous platelet inhibitors is nitric oxide (NO). NO activates NO sensitive guanylyl cyclase (NO-GC) in platelets, resulting in an increase of cyclic guanosine monophosphate (cGMP). Here we show, using cGMP sensor mice to study spatiotemporal dynamics of platelet cGMP, that NO-induced cGMP production in pre-activated platelets is strongly shear-dependent. We delineate a new mode of platelet-inhibitory mechanotransduction via shear-activated NO-GC followed by cGMP synthesis, activation of cGMP-dependent protein kinase I (cGKI), and suppression of Ca2+ signaling. Correlative profiling of cGMP dynamics and thrombus formation in vivo indicates that high cGMP concentrations in shear-exposed platelets at the thrombus periphery limit thrombosis, primarily through facilitation of thrombus dissolution. We propose that an increase in shear stress during thrombus growth activates the NO-cGMP-cGKI pathway, which acts as an auto-regulatory brake to prevent vessel occlusion, while preserving wound closure under low shear.}, language = {en} } @article{WelzEickhoffAbdullahetal.2018, author = {Welz, M. and Eickhoff, S. and Abdullah, Z. and Trebicka, J. and Gartlan, K. H. and Spicer, J. A. and Demetris, A. J. and Akhlaghi, H. and Anton, M. and Manske, K. and Zehn, D. and Nieswandt, B. and Kurts, C. and Trapani, J. A. and Knolle, P. and Wohlleber, D. and Kastenm{\"u}ller, W.}, title = {Perforin inhibition protects from lethal endothelial damage during fulminant viral hepatitis}, series = {Nature Communications}, volume = {9}, journal = {Nature Communications}, doi = {10.1038/s41467-018-07213-x}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233593}, year = {2018}, abstract = {CD8 T cells protect the liver against viral infection, but can also cause severe liver damage that may even lead to organ failure. Given the lack of mechanistic insights and specific treatment options in patients with acute fulminant hepatitis, we develop a mouse model reflecting a severe acute virus-induced CD8 T cell-mediated hepatitis. Here we show that antigen-specific CD8 T cells induce liver damage in a perforin-dependent manner, yet liver failure is not caused by effector responses targeting virus-infected hepatocytes alone. Additionally, CD8 T cell mediated elimination of cross-presenting liver sinusoidal endothelial cells causes endothelial damage that leads to a dramatically impaired sinusoidal perfusion and indirectly to hepatocyte death. With the identification of perforin-mediated killing as a critical pathophysiologic mechanism of liver failure and the protective function of a new class of perforin inhibitor, our study opens new potential therapeutic angles for fulminant viral hepatitis.}, language = {en} } @article{WalkerMavrommatisWardelletal.2019, author = {Walker, Brian A. and Mavrommatis, Konstantinos and Wardell, Christopher P. and Ashby, T. Cody and Bauer, Michael and Davies, Faith and Rosenthal, Adam and Wang, Hongwei and Qu, Pingping and Hoering, Antje and Samur, Mehmet and Towfic, Fadi and Ortiz, Maria and Flynt, Erin and Yu, Zhinuan and Yang, Zhihong and Rozelle, Dan and Obenauer, John and Trotter, Matthew and Auclair, Daniel and Keats, Jonathan and Bolli, Niccolo and Fulciniti, Mariateresa and Szalat, Raphael and Moreau, Phillipe and Durie, Brian and Stewart, A. Keith and Goldschmidt, Hartmut and Raab, Marc S. and Einsele, Hermann and Sonneveld, Pieter and San Miguel, Jesus and Lonial, Sagar and Jackson, Graham H. and Anderson, Kenneth C. and Avet-Loiseau, Herve and Munshi, Nikhil and Thakurta, Anjan and Morgan, Gareth}, title = {A high-risk, Double-Hit, group of newly diagnosed myeloma identified by genomic analysis}, series = {Leukemia}, volume = {33}, journal = {Leukemia}, doi = {10.1038/s41375-018-0196-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233299}, pages = {159-170}, year = {2019}, abstract = {Patients with newly diagnosed multiple myeloma (NDMM) with high-risk disease are in need of new treatment strategies to improve the outcomes. Multiple clinical, cytogenetic, or gene expression features have been used to identify high-risk patients, each of which has significant weaknesses. Inclusion of molecular features into risk stratification could resolve the current challenges. In a genome-wide analysis of the largest set of molecular and clinical data established to date from NDMM, as part of the Myeloma Genome Project, we have defined DNA drivers of aggressive clinical behavior. Whole-genome and exome data from 1273 NDMM patients identified genetic factors that contribute significantly to progression free survival (PFS) and overall survival (OS) (cumulative R2 = 18.4\% and 25.2\%, respectively). Integrating DNA drivers and clinical data into a Cox model using 784 patients with ISS, age, PFS, OS, and genomic data, the model has a cumlative R2 of 34.3\% for PFS and 46.5\% for OS. A high-risk subgroup was defined by recursive partitioning using either a) bi-allelic TP53 inactivation or b) amplification (≥4 copies) of CKS1B (1q21) on the background of International Staging System III, comprising 6.1\% of the population (median PFS = 15.4 months; OS = 20.7 months) that was validated in an independent dataset. Double-Hit patients have a dire prognosis despite modern therapies and should be considered for novel therapeutic approaches.}, language = {en} } @article{WegertVokuhlCollordetal.2018, author = {Wegert, Jenny and Vokuhl, Christian and Collord, Grace and Del Castillo Velasco-Herrera, Martin and Farndon, Sarah J. and Guzzo, Charlotte and Jorgensen, Mette and Anderson, John and Slater, Olga and Duncan, Catriona and Bausenwein, Sabrina and Streitenberger, Heike and Ziegler, Barbara and Furtw{\"a}ngler, Rhoikos and Graf, Norbert and Stratton, Michael R. and Campbell, Peter J. and Jones, David TW and Koelsche, Christian and Pfister, Stefan M. and Mifsud, William and Sebire, Neil and Sparber-Sauer, Monika and Koscielniak, Ewa and Rosenwald, Andreas and Gessler, Manfred and Behjati, Sam}, title = {Recurrent intragenic rearrangements of EGFR and BRAF in soft tissue tumors of infants}, series = {Nature Communications}, volume = {9}, journal = {Nature Communications}, doi = {10.1038/s41467-018-04650-6}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233446}, year = {2018}, abstract = {Soft tissue tumors of infancy encompass an overlapping spectrum of diseases that pose unique diagnostic and clinical challenges. We studied genomes and transcriptomes of cryptogenic congenital mesoblastic nephroma (CMN), and extended our findings to five anatomically or histologically related soft tissue tumors: infantile fibrosarcoma (IFS), nephroblastomatosis, Wilms tumor, malignant rhabdoid tumor, and clear cell sarcoma of the kidney. A key finding is recurrent mutation of EGFR in CMN by internal tandem duplication of the kinase domain, thus delineating CMN from other childhood renal tumors. Furthermore, we identify BRAF intragenic rearrangements in CMN and IFS. Collectively these findings reveal novel diagnostic markers and therapeutic strategies and highlight a prominent role of isolated intragenic rearrangements as drivers of infant tumors.}, language = {en} } @article{AnanyKreckelFuellsacketal.2018, author = {Anany, Mohamed A. and Kreckel, Jennifer and F{\"u}llsack, Simone and Rosenthal, Alevtina and Otto, Christoph and Siegmund, Daniela and Wajant, Harald}, title = {Soluble TNF-like weak inducer of apoptosis (TWEAK) enhances poly(I:C)-induced RIPK1-mediated necroptosis}, series = {Cell Death \& Disease}, volume = {9}, journal = {Cell Death \& Disease}, doi = {10.1038/s41419-018-1137-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-221104}, year = {2018}, abstract = {TNF-like weak inducer of apoptosis (TWEAK) and inhibition of protein synthesis with cycloheximide (CHX) sensitize for poly(I:C)-induced cell death. Notably, although CHX preferentially enhanced poly(I:C)-induced apoptosis, TWEAK enhanced primarily poly(I:C)-induced necroptosis. Both sensitizers of poly(I:C)-induced cell death, however, showed no major effect on proinflammatory poly(I:C) signaling. Analysis of a panel of HeLa-RIPK3 variants lacking TRADD, RIPK1, FADD, or caspase-8 expression revealed furthermore similarities and differences in the way how poly(I:C)/TWEAK, TNF, and TRAIL utilize these molecules for signaling. RIPK1 turned out to be essential for poly(I:C)/TWEAK-induced caspase-8-mediated apoptosis but was dispensable for this response in TNF and TRAIL signaling. TRADD-RIPK1-double deficiency differentially affected poly(I:C)-triggered gene induction but abrogated gene induction by TNF completely. FADD deficiency abrogated TRAIL- but not TNF- and poly(I:C)-induced necroptosis, whereas TRADD elicited protective activity against all three death inducers. A general protective activity against poly(I:C)-, TRAIL-, and TNF-induced cell death was also observed in FLIPL and FLIPS transfectrants.}, language = {en} } @article{LanghauserCasasDaoetal.2018, author = {Langhauser, Friederike and Casas, Ana I. and Dao, Vu-Thao-Vi and Guney, Emre and Menche, J{\"o}rg and Geuss, Eva and Kleikers, Pamela W. M. and L{\´o}pez, Manuela G. and Barab{\´a}si, Albert-L. and Kleinschnitz, Christoph and Schmidt, Harald H. H. W.}, title = {A diseasome cluster-based drug repurposing of soluble guanylate cyclase activators from smooth muscle relaxation to direct neuroprotection}, series = {npj Systems Biology and Applications}, volume = {4}, journal = {npj Systems Biology and Applications}, doi = {10.1038/s41540-017-0039-7}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236381}, year = {2018}, abstract = {Network medicine utilizes common genetic origins, markers and co-morbidities to uncover mechanistic links between diseases. These links can be summarized in the diseasome, a comprehensive network of disease-disease relationships and clusters. The diseasome has been influential during the past decade, although most of its links are not followed up experimentally. Here, we investigate a high prevalence unmet medical need cluster of disease phenotypes linked to cyclic GMP. Hitherto, the central cGMP-forming enzyme, soluble guanylate cyclase (sGC), has been targeted pharmacologically exclusively for smooth muscle modulation in cardiology and pulmonology. Here, we examine the disease associations of sGC in a non-hypothesis based manner in order to identify possibly previously unrecognized clinical indications. Surprisingly, we find that sGC, is closest linked to neurological disorders, an application that has so far not been explored clinically. Indeed, when investigating the neurological indication of this cluster with the highest unmet medical need, ischemic stroke, pre-clinically we find that sGC activity is virtually absent post-stroke. Conversely, a heme-free form of sGC, apo-sGC, was now the predominant isoform suggesting it may be a mechanism-based target in stroke. Indeed, this repurposing hypothesis could be validated experimentally in vivo as specific activators of apo-sGC were directly neuroprotective, reduced infarct size and increased survival. Thus, common mechanism clusters of the diseasome allow direct drug repurposing across previously unrelated disease phenotypes redefining them in a mechanism-based manner. Specifically, our example of repurposing apo-sGC activators for ischemic stroke should be urgently validated clinically as a possible first-in-class neuroprotective therapy.}, language = {en} } @article{LiaoTtofaliSlotkowskietal.2019, author = {Liao, Chunyu and Ttofali, Fani and Slotkowski, Rebecca A. and Denny, Steven R. and Cecil, Taylor D. and Leenay, Ryan T. and Keung, Albert J. and Beisel, Chase L.}, title = {Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis}, series = {Nature Communications}, volume = {10}, journal = {Nature Communications}, doi = {10.1038/s41467-019-10747-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236843}, year = {2019}, abstract = {CRISPR-Cas systems inherently multiplex through CRISPR arrays—whether to defend against different invaders or mediate multi-target editing, regulation, imaging, or sensing. However, arrays remain difficult to generate due to their reoccurring repeat sequences. Here, we report a modular, one-pot scheme called CRATES to construct CRISPR arrays and array libraries. CRATES allows assembly of repeat-spacer subunits using defined assembly junctions within the trimmed portion of spacers. Using CRATES, we construct arrays for the single-effector nucleases Cas9, Cas12a, and Cas13a that mediated multiplexed DNA/RNA cleavage and gene regulation in cell-free systems, bacteria, and yeast. CRATES further allows the one-pot construction of array libraries and composite arrays utilized by multiple Cas nucleases. Finally, array characterization reveals processing of extraneous CRISPR RNAs from Cas12a terminal repeats and sequence- and context-dependent loss of RNA-directed nuclease activity via global RNA structure formation. CRATES thus can facilitate diverse multiplexing applications and help identify factors impacting crRNA biogenesis.}, language = {en} } @article{LevyBoulleEmeritetal.2019, author = {Levy, Marion J. F. and Boulle, Fabien and Emerit, Michel Boris and Poilbout, Corinne and Steinbusch, Harry W. M. and Van den Hove, Daniel L. A. and Kenis, Gunter and Lanfumey, Laurence}, title = {5-HTT independent effects of fluoxetine on neuroplasticity}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-42775-w}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236759}, year = {2019}, abstract = {Selective serotonin reuptake inhibitors are among the most prescribed antidepressants. Fluoxetine is the lead molecule which exerts its therapeutic effects, at least in part, by promoting neuroplasticity through increased brain-derived neurotrophic factor (BDNF)/tropomyosin-related receptor kinase B (TrkB) signalling. It is unclear however, to which extent the neuroplastic effects of fluoxetine are solely mediated by the inhibition of the serotonin transporter (5-HTT). To answer this question, the effects of fluoxetine on neuroplasticity were analysed in both wild type (WT) and 5-Htt knock-out (KO) mice. Using Western blotting and RT-qPCR approaches, we showed that fluoxetine 10 µM activated BDNF/TrkB signalling pathways in both CD1 and C57BL/6J mouse primary cortical neurons. Interestingly, effects on BDNF signalling were observed in primary cortical neurons from both 5-Htt WT and KO mice. In addition, a 3-week in vivo fluoxetine treatment (15 mg/kg/d; i.p.) increased the expression of plasticity genes in brains of both 5-Htt WT and KO mice, and tended to equally enhance hippocampal cell proliferation in both genotypes, without reaching significance. Our results further suggest that fluoxetine-induced neuroplasticity does not solely depend on 5-HTT blockade, but might rely, at least in part, on 5-HTT-independent direct activation of TrkB.}, language = {en} } @article{KurabiSchaererNoacketal.2018, author = {Kurabi, Arwa and Schaerer, Daniel and Noack, Volker and Bernhardt, Marlen and Pak, Kwang and Alexander, Thomas and Husseman, Jacob and Nguyen, Quyen and Harris, Jeffrey P. and Ryan, Allen F.}, title = {Active Transport of Peptides Across the Intact Human Tympanic Membrane}, series = {Scientific Reports}, volume = {8}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-30031-6}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-230929}, year = {2018}, abstract = {We previously identified peptides that are actively transported across the intact tympanic membrane (TM) of rats with infected middle ears. To assess the possibility that this transport would also occur across the human TM, we first developed and validated an assay to evaluate transport in vitro using fragments of the TM. Using this assay, we demonstrated the ability of phage bearing a TM-transiting peptide to cross freshly dissected TM fragments from infected rats or from uninfected rats, guinea pigs and rabbits. We then evaluated transport across fragments of the human TM that were discarded during otologic surgery. Human trans-TM transport was similar to that seen in the animal species. Finally, we found that free peptide, unconnected to phage, was transported across the TM at a rate comparable to that seen for peptide-bearing phage. These studies provide evidence supporting the concept of peptide-mediated drug delivery across the intact TM and into the middle ears of patients.}, language = {en} } @article{LopezKleinheinzAukemaetal.2019, author = {L{\´o}pez, Cristina and Kleinheinz, Kortine and Aukema, Sietse M. and Rohde, Marius and Bernhart, Stephan H. and H{\"u}bschmann, Daniel and Wagener, Rabea and Toprak, Umut H. and Raimondi, Francesco and Kreuz, Markus and Waszak, Sebastian M. and Huang, Zhiqin and Sieverling, Lina and Paramasivam, Nagarajan and Seufert, Julian and Sungalee, Stephanie and Russell, Robert B. and Bausinger, Julia and Kretzmer, Helene and Ammerpohl, Ole and Bergmann, Anke K. and Binder, Hans and Borkhardt, Arndt and Brors, Benedikt and Claviez, Alexander and Doose, Gero and Feuerbach, Lars and Haake, Andrea and Hansmann, Martin-Leo and Hoell, Jessica and Hummel, Michael and Korbel, Jan O. and Lawerenz, Chris and Lenze, Dido and Radlwimmer, Bernhard and Richter, Julia and Rosenstiel, Philip and Rosenwald, Andreas and Schilhabel, Markus B. and Stein, Harald and Stilgenbauer, Stephan and Stadler, Peter F. and Szczepanowski, Monika and Weniger, Marc A. and Zapatka, Marc and Eils, Roland and Lichter, Peter and Loeffler, Markus and M{\"o}ller, Peter and Tr{\"u}mper, Lorenz and Klapper, Wolfram and Hoffmann, Steve and K{\"u}ppers, Ralf and Burkhardt, Birgit and Schlesner, Matthias and Siebert, Reiner}, title = {Genomic and transcriptomic changes complement each other in the pathogenesis of sporadic Burkitt lymphoma}, series = {Nature Communications}, volume = {10}, journal = {Nature Communications}, organization = {ICGC MMML-Seq Consortium}, doi = {10.1038/s41467-019-08578-3}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-237281}, year = {2019}, abstract = {Burkitt lymphoma (BL) is the most common B-cell lymphoma in children. Within the International Cancer Genome Consortium (ICGC), we performed whole genome and transcriptome sequencing of 39 sporadic BL. Here, we unravel interaction of structural, mutational, and transcriptional changes, which contribute to MYC oncogene dysregulation together with the pathognomonic IG-MYC translocation. Moreover, by mapping IGH translocation breakpoints, we provide evidence that the precursor of at least a subset of BL is a B-cell poised to express IGHA. We describe the landscape of mutations, structural variants, and mutational processes, and identified a series of driver genes in the pathogenesis of BL, which can be targeted by various mechanisms, including IG-non MYC translocations, germline and somatic mutations, fusion transcripts, and alternative splicing.}, language = {en} } @article{LuBoswellBoswelletal.2019, author = {Lu, Yuan and Boswell, Wiliam and Boswell, Mikki and Klotz, Barbara and Kneitz, Susanne and Regneri, Janine and Savage, Markita and Mendoza, Cristina and Postlethwait, John and Warren, Wesley C. and Schartl, Manfred and Walter, Ronald B.}, title = {Application of the Transcriptional Disease Signature (TDSs) to Screen Melanoma-Effective Compounds in a Small Fish Model}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-36656-x}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-237322}, year = {2019}, abstract = {Cell culture and protein target-based compound screening strategies, though broadly utilized in selecting candidate compounds, often fail to eliminate candidate compounds with non-target effects and/or safety concerns until late in the drug developmental process. Phenotype screening using intact research animals is attractive because it can help identify small molecule candidate compounds that have a high probability of proceeding to clinical use. Most FDA approved, first-in-class small molecules were identified from phenotypic screening. However, phenotypic screening using rodent models is labor intensive, low-throughput, and very expensive. As a novel alternative for small molecule screening, we have been developing gene expression disease profiles, termed the Transcriptional Disease Signature (TDS), as readout of small molecule screens for therapeutic molecules. In this concept, compounds that can reverse, or otherwise affect known disease-associated gene expression patterns in whole animals may be rapidly identified for more detailed downstream direct testing of their efficacy and mode of action. To establish proof of concept for this screening strategy, we employed a transgenic strain of a small aquarium fish, medaka (Oryzias latipes), that overexpresses the malignant melanoma driver gene xmrk, a mutant egfr gene, that is driven by a pigment cell-specific mitf promoter. In this model, melanoma develops with 100\% penetrance. Using the transgenic medaka malignant melanoma model, we established a screening system that employs the NanoString nCounter platform to quantify gene expression within custom sets of TDS gene targets that we had previously shown to exhibit differential transcription among xmrk-transgenic and wild-type medaka. Compound-modulated gene expression was identified using an internet-accessible custom-built data processing pipeline. The effect of a given drug on the entire TDS profile was estimated by comparing compound-modulated genes in the TDS using an activation Z-score and Kolmogorov-Smirnov statistics. TDS gene probes were designed that target common signaling pathways that include proliferation, development, toxicity, immune function, metabolism and detoxification. These pathways may be utilized to evaluate candidate compounds for potential favorable, or unfavorable, effects on melanoma-associated gene expression. Here we present the logistics of using medaka to screen compounds, as well as, the development of a user-friendly NanoString data analysis pipeline to support feasibility of this novel TDS drug-screening strategy.}, language = {en} } @article{MeralProvasiPradaGraciaetal.2018, author = {Meral, Derya and Provasi, Davide and Prada-Gracia, Diego and M{\"o}ller, Jan and Marino, Kristen and Lohse, Martin J. and Filizola, Marta}, title = {Molecular details of dimerization kinetics reveal negligible populations of transient µ-opioid receptor homodimers at physiological concentrations}, series = {Scientific Reports}, volume = {8}, journal = {Scientific Reports}, doi = {10.1038/s41598-018-26070-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223995}, year = {2018}, abstract = {Various experimental and computational techniques have been employed over the past decade to provide structural and thermodynamic insights into G Protein-Coupled Receptor (GPCR) dimerization. Here, we use multiple microsecond-long, coarse-grained, biased and unbiased molecular dynamics simulations (a total of ~4 milliseconds) combined with multi-ensemble Markov state models to elucidate the kinetics of homodimerization of a prototypic GPCR, the µ-opioid receptor (MOR), embedded in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/cholesterol lipid bilayer. Analysis of these computations identifies kinetically distinct macrostates comprising several different short-lived dimeric configurations of either inactive or activated MOR. Calculated kinetic rates and fractions of dimers at different MOR concentrations suggest a negligible population of MOR homodimers at physiological concentrations, which is supported by acceptor photobleaching fluorescence resonance energy transfer (FRET) experiments. This study provides a rigorous, quantitative explanation for some conflicting experimental data on GPCR oligomerization.}, language = {en} } @article{MedlerNelkeWeisenbergeretal.2019, author = {Medler, Juliane and Nelke, Johannes and Weisenberger, Daniela and Steinfatt, Tim and Rothaug, Moritz and Berr, Susanne and H{\"u}nig, Thomas and Beilhack, Andreas and Wajant, Harald}, title = {TNFRSF receptor-specific antibody fusion proteins with targeting controlled FcγR-independent agonistic activity}, series = {Cell Death \& Disease}, volume = {10}, journal = {Cell Death \& Disease}, doi = {10.1038/s41419-019-1456-x}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223948}, year = {2019}, abstract = {Antibodies specific for TNFRSF receptors that bind soluble ligands without getting properly activated generally act as strong agonists upon FcγR binding. Systematic analyses revealed that the FcγR dependency of such antibodies to act as potent agonists is largely independent from isotype, FcγR type, and of the epitope recognized. This suggests that the sole cellular attachment, achieved by Fc domain-FcγR interaction, dominantly determines the agonistic activity of antibodies recognizing TNFRSF receptors poorly responsive to soluble ligands. In accordance with this hypothesis, we demonstrated that antibody fusion proteins harboring domains allowing FcγR-independent cell surface anchoring also act as strong agonist provided they have access to their target. This finding defines a general possibility to generate anti-TNFRSF receptor antibodies with FcγR-independent agonism. Moreover, anti-TNFRSF receptor antibody fusion proteins with an anchoring domain promise superior applicability to conventional systemically active agonists when an anchoring target with localized disease associated expression can be addressed.}, language = {en} } @phdthesis{Jorgacevic2024, author = {Jorgacevic, Ivana}, title = {Elucidating the interconnection of GvHD and Western diet-induced atherosclerosis}, doi = {10.25972/OPUS-32579}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-325792}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Allogeneic hematopoietic cell transplantation (Allo-HCT) is the main and only treatment for many malignant and non-malignant haematological disorders. Even though the treatment has improved through the years and patient life expectancy has increased, graft versus host disease (GvHD) is still considered the main obstacle and one of the main reasons for increased mortality. Furthermore, improved patient's survival and life expectancy brought into question the late post-HCT complications. The leading cause of late death after allo-HCT is the relapse of primary disease as well as chronic GvHD (cGvHD). However, a clear relationship was also described with pulmonary complications, endocrine dysfunction and infertility, and cataracts in post-HCT patients. In the last years big concern regarding a cumulative cardiovascular incidence in long-term survivors has been raised. Severe cardiovascular disease (CVD) is caused by atherosclerosis which is considered a chronic inflammatory disease of blood vessels. As such, it takes a long time from endothelial damage, as the onset event, and followed plaque formation to a manifestation of severe consequences, such as stroke, coronary heart disease, or peripheral arterial disease. Endothelial damage is well documented in patients post-HCT. In the context of allo-HCT, the endothelial damage is induced by the conditioning regimen with or without total body irradiation (TBI). Furthermore, endothelial cells (ECs) have been documented as a target of GvHD and increased concentration of circulating endothelial cells (CEC) coinciding with an increase in the number of circulating alloreactive T cells. According to 2021 ESC Guidelines on CVD prevention, the main atherosclerotic CVD (ASCVD) risk factors are blood apolipoprotein B (ApoB)-containing lipoproteins (of which low-density lipoprotein (LDL) is the most abundant), high blood pressure, cigarette smoking and diabetes mellitus (DM). GvHD is considered a high-risk factor for the onset of dyslipidaemia, hypertension, and DM. Overall, the risk of premature cardiovascular death is 2.7 fold increased in comparison to the general population, while the cumulative incidence of cardiovascular complications was shown to be up to 47\% at ten years after reduced intensity conditioning (RIC), post-HCT. However, up to date, there are no available studies elucidating the interconnection between GvHD and atherosclerosis. The goal of this study was, therefore, to investigate the involvement of GvHD in the progression of atherosclerosis as well as to elucidate whether cytotoxic, CD8+ T cells that were shown to play a significant role in endothelial damage during the course of skin GvHD on one hand, and inducers of formation of unstable plaque on the other, are involved in this interconnection. For that purpose we established a novel minor histocompatibility anti gens (miHAg) allo-HCT Western diet (WD)-induced atherosclerosis mouse model. We were able to show that GvHD has a significant impact on atherosclerosis development in B6.Ldlr-/- recipient mice even in the absence of overt clinical disease activity. It seems that the impact is at least partly induced by CD8+ T cells, that showed significantly increased infiltration of aortic lesions in mice facing subclinical GvHD. As studies have shown in regular atherosclerotic mouse models as well as in humans, these CD8+ T cells exhibited not only increased expression of genes involved in activation, survival and differentiation to cytotoxic phenotype, but also some genes pointing out their exhaustion, that were absent in CD4+ T cell cluster. When anti-CD8β antibody was applied once per week along with WD feeding for eight weeks, the plaque formation was significantly reduced in aorta and aortic root pointing out the importance of these cells in an alloreactivity induced lesion formation. Furthermore, anti-CD8β treatment led to significantly decreased necrotic core formation followed by overall increase in plaque stability. Strikingly, bone marrow plus T cells (BMT) recipients fed WD showed significantly increased serum cholesterol levels in comparison to bone marrow (BM) (a group lacking alloreactive T cells that induce GvHD). This effect was reversed when anti-CD8β treatment was applied, suggesting, at least partly, an impact of alloreactive CD8+ T cells on cholesterol levels. Expression of genes responsible for lipid metabolism pointed out the tendency of the liver to regulate the increased cholesterol levels, however, the mechanism behind this phenotype still remains to be revealed. On the other hand, the impact of obesity, induced by chronic high-fat diet (HFD) feeding, has been shown to be an independent risk factor for gastrointestinal GvHD. Similarly, in major histocompatibility complex (MHC) disparate allo-HCT mouse model, we have noticed that even short-term WD intake leads to a significant decrease in survival of mice post-HCT. When the concentration of transplanted alloreactive T cells was reduced, the survival was improved, pointing out the involvement of these cells in the pathogenesis. Additionally, bioluminescence imaging (BLI) during initiation and effector phase of acute GvHD (aGvHD) revealed increased infiltration of alloreactive T cells in mice fed WD. Studies in an obesity model, we could confirm the involvement of specifically CD4+ T cells in WD induced impact, as the relative number of these cells was significantly increased in small intestine on day six post-HCT in mice fed WD. This increased intestinal infiltration was preceded by increase in the number of alloreactive T cells expressing intestine homing receptor (α4β7 integrin) in peripheral lymph nodes (LNs). Even though the number of T cells was not changed in the spleen of WD fed mice, the subset of CD4+ and CD8+ T cells that were highly secreting TNFα was increased as well as the expression of genes regulating pro-inflammatory cytokines such as IL-6 and interferon (IFN)γ pointing out significant WD-induced inflammation. Moreover, slight tendency towards increased intestinal permeability and load of translocated luminal bacteria, that we observed, could induce severe endotoxemia and dysregulated systemic immune response that could lead to detrimental induction of cell death. Justifying our speculations, we noted increased levels of transaminases and an increase in lactate dehydrogenase (LDH) levels (pointing out significant tissue damages). However, the exact mechanism behind this detrimental WD impact still remains to be elucidated.}, subject = {Periphere Stammzellentransplantation}, language = {en} } @phdthesis{Papay2024, author = {Papay, Marion}, title = {Notwendigkeit der pr{\"a}operativen Reposition von distalen, nach dorsal dislozierten Radiusfrakturen bei bestehender Operationsindikation im Hinblick auf das Schmerzniveau sowie postoperative Ergebnisse}, doi = {10.25972/OPUS-36388}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-363882}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die distale Radiusfraktur geh{\"o}rt zu den h{\"a}ufigsten Frakturen in Deutschland mit einem Inzidenzanstieg im Alter unter Betonung des weiblichen Geschlechts. Dabei zeigt sich ein zunehmender Trend in Richtung operative Versorgung, allen voran die Versorgung mittels winkelstabiler Plattensysteme. Instabile, distale Radiusfrakturen werden dabei vor geplanter operativer Versorgung im Rahmen der Initialbehandlung {\"u}blicherweise geschlossen reponiert und im Gipsverband retiniert. Ziel der vorliegenden monozentrischen, prospektiv randomisierten Studie mit zwei Studiengruppen war es herauszufinden, ob sich das Unterlassen der Reposition vor geplanter Operation nachteilig auf das Schmerzniveau in der pr{\"a}operativen Phase auswirkt und ob sich durch die Dislokation Nachteile in Bezug auf den Nervus medianus im Sinne eines Traktionsschadens sowie bez{\"u}glich des klinisch-radiologischen Ausheilungsergebnisses zeigen. Die Studie zeigte, dass das Schmerzempfinden w{\"a}hrend der pr{\"a}operativen Gipsbehandlung unabh{\"a}ngig von einer vorherigen Reposition war. F{\"u}r den prim{\"a}ren Endpunkt an Tag 1 nach der Akutbehandlung konnte statistisch signifikante Nichtunterlegenheit der Gruppe ohne Reposition gegen{\"u}ber der Gruppe mit Reposition nachgewiesen werden. Gleiches galt f{\"u}r Tag 2, sowohl f{\"u}r die absoluten Schmerzniveaus als auch f{\"u}r die Schmerzlinderung. Das Unterlassen der Reposition hatte zudem keine nachteiligen Effekte auf den Nervus medianus. Gleiches zeigte sich f{\"u}r das klinische und radiologische Ausheilungsergebnis. F{\"u}r die funktionellen DASH- und Krimmer-Scores konnte ein Jahr postoperativ ebenfalls statistisch signifikante Nichtunterlegenheit der Gruppe ohne Reposition nachgewiesen werden. Diese Erkenntnisse best{\"a}tigen die in der Literatur vorhandenen Ergebnisse verschiedener Studien dahingehend, dass das Unterlassen der Reposition keine nachteiligen Effekte auf das postoperative Outcome hat. Einige Studien verdeutlichen zudem, dass es nach Reposition, insbesondere bei Vorliegen gewisser Risiko- und Instabilit{\"a}tsfaktoren, ohnehin zur sekund{\"a}ren Dislokation kommt, sodass die generelle Notwendigkeit der Reposition vor Gipsanlage sowohl vor einer operativen als auch vor einer konservativen Weiterbehandlung angezweifelt werden muss.}, subject = {distale Radiusfraktur}, language = {de} } @phdthesis{vonderHeide2024, author = {von der Heide, Julia Magdalena}, title = {Ist eine Berechnung der Geometrie der Halswirbelk{\"o}rper anhand ihrer Morphologie im Kindesalter und somit eine Individualisierung der CVM-Methode m{\"o}glich?}, doi = {10.25972/OPUS-36075}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-360753}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Ziel der vorliegenden Studie war es zu untersuchen, ob anhand geometrischer Merkmale der HWK im Kindesalter eine sichere individuelle Vorhersage der Morphologie der HWK zum Ende der Entwicklung m{\"o}glich ist. Hierdurch k{\"o}nnte eine Individualisierung der CVM-Methode und somit eine Einsch{\"a}tzung des bereits verstrichenen Wachstums erfolgen. Zu diesem Zweck wurden insgesamt 1377 FRS-Aufnahmen von 267 Patienten - 110 weibliche und 157 m{\"a}nnliche - aus dem Archiv der Poliklinik f{\"u}r Kieferorthop{\"a}die des Universit{\"a}tsklinikums W{\"u}rzburg digitalisiert und untersucht. Die HWK wurden im Programm OnyxCeph (Herst.: Image Instruments GmbH) quantifiziert und die berechneten Werte mit der Software SPSS statistisch ausgewertet. Mittels linearer Regressionen wurde versucht, anhand der Morphologie der Wirbelk{\"o}rper vor dem puberalen Wachstumsschub auf die Geometrie der HWK im Erwachsenenalter zu schließen. Zur Illustrierung wurden Streudiagramm und die dazugeh{\"o}rigen Abfolgen von R{\"o}ntgenbildern dargestellt. Eine Sch{\"a}tzung der Geometrie der HWK im Erwachsenenalter w{\"u}rde bei den separat betrachteten Parametern und bei einer gemeinsamen Betrachtung der Parameter kaum zu korrekten Einsch{\"a}tzungen f{\"u}hren. Die Streudiagramme mit den Bilderabfolgen st{\"u}tzen diese These ebenfalls und illustrieren die m{\"o}gliche Fehleinsch{\"a}tzung der Geometrie. Die Ergebnisse der Studie zeigen erneut, dass die Geometrie der HWK im Erwachsenenalter sehr variabel ist, wie komplex die Entwicklung der HWK ist und dass anhand ihrer Geometrie im Kindesalter keine sichere Einsch{\"a}tzung der skelettalen Reife m{\"o}glich ist. Eine Individualisierung der CVM-Methode ist anhand der in dieser Studie untersuchten Parameter nicht m{\"o}glich. Somit l{\"a}sst sich schlussfolgern, dass die CVM-Methode nicht als alleinige Methode zur pr{\"a}zisen skelettalen Alterseinsch{\"a}tzung verl{\"a}sslich genutzt werden kann, sondern f{\"u}r eine sichere Beurteilung weitere Reifeindikatoren hinzugezogen werden sollten. Allerdings sollten hierzu zus{\"a}tzliche radiologische Untersuchungen, wie beispielsweise die Handr{\"o}ntgenaufnahme, nur dann durchgef{\"u}hrt werden, wenn diese dem ALARA-Prinzip entsprechen.}, subject = {Skelett}, language = {de} } @phdthesis{Krings2024, author = {Krings, Moritz}, title = {Universit{\"a}re Psychiatrie um 1900 : Die Anfangsjahre der psychiatrischen Klinik in W{\"u}rzburg}, doi = {10.25972/OPUS-36140}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-361407}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Ende des 19. Jahrhunderts standen sich in Deutschland zwei verschiedene Arten psychiatrischer Institutionen gegen{\"u}ber, die Anstaltspsychiatrien auf der einen, die universit{\"a}ren psychiatrischen Kliniken auf der anderen Seite. Die psychiatriehistorische Forschung widmete sich {\"u}berwiegend psychiatrischen Anstalten w{\"a}hrend Kliniken hier unterrepr{\"a}sentiert sind. Die vorliegende Arbeit m{\"o}chte zur historischen Kenntnis universit{\"a}rer psychiatrischer Einrichtungen beitragen. Hierzu werden die Charakteristika einer psychiatrischen Klinik um 1900 anhand des Beispiels der psychiatrischen Klinik der Universit{\"a}t W{\"u}rzburg betrachtet. Der Fokus liegt hierbei neben Lage und Aufbau der Klinik sowie deren Personal auf den drei Bereichen Patient*innen, Forschung und Lehre.}, subject = {Julius-Maximilians-Universit{\"a}t W{\"u}rzburg}, language = {de} } @phdthesis{Gaballa2024, author = {Gaballa, Abdallah Hatem Hassan Hosny Ahmed}, title = {PAF1c drives MYC-mediated immune evasion in pancreatic ductal adenocarcinoma}, doi = {10.25972/OPUS-36045}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-360459}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {The expression of the MYC proto-oncogene is elevated in a large proportion of patients with pancreatic ductal adenocarcinoma (PDAC). Previous findings in PDAC have shown that this increased MYC expression mediates immune evasion and promotes S-phase progression. How these functions are mediated and whether a downstream factor of MYC mediates these functions has remained elusive. Recent studies identifying the MYC interactome revealed a complex network of interaction partners, highlighting the need to identify the oncogenic pathway of MYC in an unbiased manner. In this work, we have shown that MYC ensures genomic stability during S-phase and prevents transcription-replication conflicts. Depletion of MYC and inhibition of ATR kinase showed a synergistic effect to induce DNA damage. A targeted siRNA screen targeting downstream factors of MYC revealed that PAF1c is required for DNA repair and S-phase progression. Recruitment of PAF1c to RNAPII was shown to be MYC dependent. PAF1c was shown to be largely dispensable for cell proliferation and regulation of MYC target genes. Depletion of CTR9, a subunit of PAF1c, caused strong tumor regression in a pancreatic ductal adenocarcinoma model, with long-term survival in a subset of mice. This effect was not due to induction of DNA damage, but to restoration of tumor immune surveillance. Depletion of PAF1c resulted in the release of RNAPII with transcription elongation factors, including SPT6, from the bodies of long genes, promoting full-length transcription of short genes. This resulted in the downregulation of long DNA repair genes and the concomitant upregulation of short genes, including MHC class I genes. These data demonstrate that a balance between long and short gene transcription is essential for tumor progression and that interference with PAF1c levels shifts this balance toward a tumor-suppressive transcriptional program. It also directly links MYC-mediated S-phase progression to immune evasion. Unlike MYC, PAF1c has a stable, known folded structure; therefore, the development of a small molecule targeting PAF1c may disrupt the immune evasive function of MYC while sparing its physiological functions in cellular growth.}, subject = {Myc}, language = {en} } @phdthesis{Polzin2024, author = {Polzin, Charlotte}, title = {Entwicklung eines Screeningverfahrens f{\"u}r Linezolid-resistente Enterokokken und Aufnahme der Pr{\"a}valenz}, doi = {10.25972/OPUS-37066}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-370665}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Enterokokken geh{\"o}ren zu den bedeutendsten nosokomialen Keimen. Die Verbreitung von Multiresistenzen bei diesen Keimen stellt das deutsche Gesundheitssystem aufgrund von wenigen verbleibenden Therapieoptionen von Infektionen vor große Probleme. Die KRINKO des Robert-Koch-Instituts empfiehlt als m{\"o}gliche Pr{\"a}ventionsmaßnahme ein regelm{\"a}ßiges Screening auf Enterokokken mit Vancomycin- bzw. Linezolid-Resistenzen. Ziel dieser Arbeit war es, ein kulturelles Screeningverfahren f{\"u}r Linezolid-resistente Enterokokken (LRE) zu entwickeln und dieses anschließend im Routinescreening des Universit{\"a}tsklinikums W{\"u}rzburg zu etablieren. Es wurde ein Verfahren entwickelt, welches sich aus einem Anreicherungsschritt mit 3 mg/l Linezolid versetzter selektiver Enterococcosel-Bouillon und einer anschließenden Subkultivierung auf Linezolid-Enterococcosel-Agar mit 4 mg/l Linezolid zusammensetzt. In einer Simulation von klinischen Bedingungen zeigte sich eine gute Sensitivit{\"a}t und Spezifit{\"a}t. Das entwickelte Screeningverfahren wurde mit einem geringen Sensitivit{\"a}tsverlust und ohne zus{\"a}tzliche Belastung f{\"u}r die Patienten in das bestehende Routinescreening f{\"u}r Vancomycin-resistente Enterokokken des Universit{\"a}tsklinikums W{\"u}rzburg eingegliedert. Die nachgewiesen LRE zeigten unterschiedliche Resistenzmechanismen, wobei bei dem Großteil der Isolate Resistenzgene nachgewiesen werden konnten. Des Weiteren zeigte sich ein breit gestreuter genetischer Hintergrund. Viele der Isolate geh{\"o}rten genetischen Gruppen an, welche bisher kaum in hospitalisierten Patienten nachgewiesen wurden. Durch die labortechnische Weiterentwicklung von Screeningverfahren f{\"u}r LRE k{\"o}nnen diese m{\"o}glicherweise bald routinem{\"a}ßig in vielen Kliniken etabliert werden.}, subject = {Enterococcus}, language = {de} } @article{ElliotGermainHilzetal.2019, author = {Elliot, Perry M. and Germain, Dominique P. and Hilz, Max J. and Spada, Marco and Wanner, Christoph and Falissard, Bruno}, title = {Why systematic literature reviews in Fabry disease should include all published evidence}, series = {European Journal of Medical Genetics}, volume = {62}, journal = {European Journal of Medical Genetics}, doi = {10.1016/j.ejmg.2019.103702}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-226654}, year = {2019}, abstract = {Fabry disease is an X-linked inherited, progressive disorder of lipid metabolism resulting from the deficient activity of the enzyme α-galactosidase. Enzyme replacement therapy (ERT) with recombinant agalsidase, with intravenous infusions of either agalsidase beta or agalsidase alfa, is available and clinical experience now exceeds 15 years. There are very few randomised, placebo-controlled clinical trials evaluating the outcomes of ERT. Data are often derived from observational, registry-based studies and case reports. Pooled analysis of data from different sources may be limited by the heterogeneity of the patient populations, outcomes and treatment. Therefore, comprehensive systematic literature reviews of unpooled data are needed to determine the effects of ERT on disease outcomes. A systematic literature search was conducted in the Embase and PubMed (MEDLINE) databases to retrieve original articles that evaluated outcomes of ERT in patients with Fabry disease; the outcome data were analysed unpooled. The literature analysis included the full range of published literature including observational studies and case series/case reports. Considerable heterogeneity was found among the studies, with differences in sample size, statistical methods, ERT regimens and patient demographic and clinical characteristics. We have demonstrated the value of performing an unpooled systematic literature review of all published evidence of ERT outcomes in Fabry disease, highlighting that in a rare genetic disorder like Fabry disease, which is phenotypically diverse, different patient populations can require different disease management and therapeutic goals depending on age, genotype, and disease severity/level of organ involvement. In addition, these findings are valuable to guide the design and reporting of new clinical studies.}, language = {en} } @article{SondermannUtikalEnketal.2019, author = {Sondermann, Wiebke and Utikal, Jochen Sven and Enk, Alexander H. and Schadendorf, Dirk and Klode, Joachim and Hauschild, Axel and Weichenthal, Michael and French, Lars E. and Berking, Carola and Schilling, Bastian and Haferkamp, Sebastian and Fr{\"o}hling, Stefan and von Kalle, Christof and Brinker, Titus J.}, title = {Prediction of melanoma evolution in melanocytic nevi via artificial intelligence: A call for prospective data}, series = {European Journal of Cancer}, volume = {119}, journal = {European Journal of Cancer}, doi = {10.1016/j.ejca.2019.07.009}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239263}, pages = {30-34}, year = {2019}, abstract = {Recent research revealed the superiority of artificial intelligence over dermatologists to diagnose melanoma from images. However, 30-50\% of all melanomas and more than half of those in young patients evolve from initially benign lesions. Despite its high relevance for melanoma screening, neither clinicians nor computers are yet able to reliably predict a nevus' oncologic transformation. The cause of this lies in the static nature of lesion presentation in the current standard of care, both for clinicians and algorithms. The status quo makes it difficult to train algorithms (and clinicians) to precisely assess the likelihood of a benign skin lesion to transform into melanoma. In addition, it inhibits the precision of current algorithms since 'evolution' image features may not be part of their decision. The current literature reveals certain types of melanocytic nevi (i.e. 'spitzoid' or 'dysplastic' nevi) and criteria (i.e. visible vasculature) that, in general, appear to have a higher chance to transform into melanoma. However, owing to the cumulative nature of oncogenic mutations in melanoma, a more fine-grained early morphologic footprint is likely to be detectable by an algorithm. In this perspective article, the concept of melanoma prediction is further explored by the discussion of the evolution of melanoma, the concept for training of such a nevi classifier and the implications of early melanoma prediction for clinical practice. In conclusion, the authors believe that artificial intelligence trained on prospective image data could be transformative for skin cancer diagnostics by (a) predicting melanoma before it occurs (i.e. pre-in situ) and (b) further enhancing the accuracy of current melanoma classifiers. Necessary prospective images for this research are obtained via free mole-monitoring mobile apps.}, language = {en} } @phdthesis{Danner2024, author = {Danner, Elisabeth}, title = {Systolische Ejektionszeit - Referenzwerte und Einfluss kardiovaskul{\"a}rer Risikofaktoren in einer populationsbasierten Kohortenstudie}, doi = {10.25972/OPUS-37048}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-370480}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die systolische Ejektionszeit (SET) und die pr{\"a}-Ejektionszeit (pET) sind Zeitintervalle, die sich zuverl{\"a}ssig mittels transthorakaler Echokardiographie erheben lassen und wichtige Aspekte in der kardialen Leistungsdiagnostik abbilden. Gleichwohl gibt es bislang f{\"u}r diese systolischen Zeitintervalle keine Normwerte. Die vorliegende Arbeit analysierte echokardiographische Daten von 4965 ProbandInnen der STAAB-Studie, einer umf{\"a}nglich ph{\"a}notypisierten populationsbasierten Kohortenstudie mit dem Ziel der Erstellung von Normwerten f{\"u}r die systolischen Zeitintervalle sowie der Identifizierung und Quantifizierung ihrer Determinanten mittels multivariable Regressionsanalysen. Aufgrund der starken Abh{\"a}ngigkeit der SET von der Herzfrequenz wurde die herzfrequenzkorrigierte SETc eingef{\"u}hrt, die in Anlehnung an die QTc-Zeit nach Fridericia berechnet wurde ("SET" /∛("RR-Intervall" )). Die Normwerte wurden anhand der gesunden Untergruppe (definiert durch Abwesenheit von kardiovaskul{\"a}ren Risikofaktoren oder Erkrankungen; N=966) aus STAAB generiert. Dem starken Einfluss von Geschlecht und Alter wurde dabei Rechnung getragen, indem f{\"u}r SET, SETc und pET alters- und geschlechtsspezifische Referenzwerte berechnet wurden. SETc war stark korreliert mit linksventrikul{\"a}rer Ejektionsfraktion sowie enddiastolischem Volumen und Schlagvolumen. Zudem zeigte sich ein deutlicher Zusammenhang von SETc mit spezifischen Markern der Nachlast (Ea und Ees). Aus der großen Liste der klinischen Marker, die in STAAB erhoben wurden, ergab sich eine gr{\"o}ßere Anzahl von Einflussfaktoren. Bedeutsame Determinanten der systolischen Zeitintervalle waren insbesondere das metabolische Syndrom (sowie dessen Komponenten), das Rauchverhalten und die Einnahme von β Blockern. Die Ergebnisse zeigen, dass die systolischen Zeitintervalle, insbesondere SETc, reliable Informationen {\"u}ber das kardiale Kontraktionsverhalten liefern k{\"o}nnen. Derzeit werden neue Substanzklassen untersucht, die unter anderem {\"u}ber die Modifizierung von SET wirken sollen. Hier zu nennen sind insbesondere Myosin-Aktivatoren bei Herzinsuffizienz mit reduzierter Pumpfunktion wie zum Beispiel Omecamtiv Mecarbil oder Myosin-Inhibitoren bei Hypertropher Obstruktiver Kardiomyopathie. Die hierzu laufenden Studien betrachten auch die Effekte dieser Medikamente auf die systolischen Zeitintervalle als bedeutsame Surrogate der klinischen Effekte und letztlich der Prognose. Die nun vorliegenden Normwerte dieser Zeitintervalle erlauben es, die pathologisch ver{\"a}nderten Werte bei diesen spezifischen Krankheitsbildern und klinischen Studien besser einzuordnen und zu verstehen.}, subject = {Referenzwert}, language = {de} } @phdthesis{Engelbrecht2024, author = {Engelbrecht, Elisabeth}, title = {Retrospektive Auswertung des Therapieerfolges einer PTCD-Anlage bei Insuffizienz der Pankreatojejunostomie nach Pankreaskopfresektionen}, doi = {10.25972/OPUS-37039}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-370395}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Trotz stetiger Weiterentwicklung und Verbesserungen in chirurgischen Anastomosetechniken treten postoperative Pankreasfisteln (POPF) in 20 - 25 \% der Patienten und Patientinnen als Komplikation nach partieller Pankreatikoduodenektomie (PPD) auf. Kommt es zu einer kombinierten Leckage aus Gallen- und Pankreassekret, wurde in dieser Arbeit die Definition einer komplizierten POPF (CPPF) eingef{\"u}hrt, welche eine seltene, aber schwerwiegende und gef{\"a}hrliche postoperative Komplikation darstellt. Neben einer Relaparotomie kann eine Restpankreatektomie als ultima ratio zur Beherrschung dieser schweren Komplikation notwendig werden, welche mit einer Mortalit{\"a}t von 50 \% verbunden ist. Die Internationale Studiengruppe der Pankreaschirurgie (ISGPS) entwickelte ein Klassifikationssystem f{\"u}r POPF, welches auf Abweichungen der {\"u}blichen postoperativen Behandlungsstrategie beruht. Jedoch wurden keine spezifischen Behandlungsalgorithmen bzw. Therapiekonzepte, insbesondere im Falle einer CPPF, vorgeschlagen. In dieser Arbeit soll die therapeutische Effektivit{\"a}t einer perkutanen transhepatischen Cholangiodrainage (PTCD) bei Patienten und Patientinnen mit einer CPPF evaluiert werden. Dazu wurde eine retrospektive Analyse an Patienten und Patientinnen durchgef{\"u}hrt, welche eine CPPF nach PPD entwickelten. Die Patienten und Patientinnen wurden hinsichtlich der gew{\"a}hlten Behandlungsstrategie, des Outcomes, postoperativer Komplikationen nach Clavien-Dindo-Klassifikation, des CCI (Comprehensive Complication Index), der 30- und 90-Tage-Mortalit{\"a}t sowie Restpankreatektomie, postoperativer Arrosionsblutungen und der Hospitalisierungsdauer nach Behandlung einer CPPF analysiert. Zwischen 2007 und 2018 entwickelten 56 (19,1 \%) von insgesamt 293 Patienten und Patientinnen eine relevante POPF (ISGPS Grad B/C) nach einer Pankreaskopfresektion. Darunter wurden 17 Patienten und Patientinnen mit einer komplizierten POPF (CPPF) identifiziert. 11 Patienten und Patientinnen erhielten als Behandlung eine PTCD und sechs Patienten und Patientinnen erhielten eine chirurgisch eingebrachte transhepatische Cholangiodrainage (CTCD) im Rahmen eines Revisionseingriffes. Es wurde keine Restpankreatektomie oder Reoperation nach einer initialen PTCD Therapie notwendig. In 4 von 17 F{\"a}llen kam es zu postoperativen Blutungen nach Einbringen einer transhepatischen Cholangiodrainage, der mediane CCI lag bei 44 ± 17,3, die mediane Hospitalisierungsdauer betrug 36 ± 19,2 Tage, die 30-Tage-Mortalit{\"a}t war 0 \% und die 90-Tage-Mortalit{\"a}t 17,7 \%. Es wurde kein Sterbefall in Verbindung mit einer PTCD beobachtet. Mit Hilfe dieser Studie kann gezeigt werden, dass eine PTCD eine praktikable, sichere und effektive Behandlungsoption f{\"u}r Patienten und Patientinnen mit einer CPPF bietet. Die Separierung von Galle und Pankreassaft stellt eine neuartige Behandlungsoption in ausgew{\"a}hlten Patienten und Patientinnen mit ausreichend drainierter CPPF nach PPD dar.}, subject = {Bauchspeicheldr{\"u}senkrebs}, language = {de} } @article{FrankeMicheliniAshersonetal.2018, author = {Franke, Barbara and Michelini, Giorgia and Asherson, Philip and Banaschewski, Tobias and Bilbow, Andrea and Buitelaar, Jan K. and Cormand, Bru and Faraone, Stephen V. and Ginsberg, Ylva and Haavik, Jan and Kuntsi, Jonna and Larsson, Henrik and Lesch, Klaus-Peter and Ramos-Quiroga, J. Antoni and R{\´e}thelyi, J{\´a}nos M. and Ribases, Marta and Reif, Andreas}, title = {Live fast, die young? A review on the developmental trajectories of ADHD across the lifespan}, series = {European Neuropsychopharmacology}, volume = {28}, journal = {European Neuropsychopharmacology}, doi = {10.1016/j.euroneuro.2018.08.001}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-228407}, pages = {1059-1088}, year = {2018}, abstract = {Attention-deficit/hyperactivity disorder (ADHD) is highly heritable and the most common neurodevelopmental disorder in childhood. In recent decades, it has been appreciated that in a substantial number of cases the disorder does not remit in puberty, but persists into adulthood. Both in childhood and adulthood, ADHD is characterised by substantial comorbidity including substance use, depression, anxiety, and accidents. However, course and symptoms of the disorder and the comorbidities may fluctuate and change over time, and even age of onset in childhood has recently been questioned. Available evidence to date is poor and largely inconsistent with regard to the predictors of persistence versus remittance. Likewise, the development of comorbid disorders cannot be foreseen early on, hampering preventive measures. These facts call for a lifespan perspective on ADHD from childhood to old age. In this selective review, we summarise current knowledge of the long-term course of ADHD, with an emphasis on clinical symptom and cognitive trajectories, treatment effects over the lifespan, and the development of comorbidities. Also, we summarise current knowledge and important unresolved issues on biological factors underlying different ADHD trajectories. We conclude that a severe lack of knowledge on lifespan aspects in ADHD still exists for nearly every aspect reviewed. We encourage large-scale research efforts to overcome those knowledge gaps through appropriately granular longitudinal studies.}, language = {en} } @article{EstesAnsteeAriasLosteetal.2018, author = {Estes, Chris and Anstee, Quentin M. and Arias-Loste, Maria Teresa and Bantel, Heike and Bellentani, Stefano and Caballeria, Joan and Colombo, Massimo and Craxi, Antonio and Crespo, Javier and Day, Christopher P. and Eguchi, Yuichiro and Geier, Andreas and Kondili, Loreta A. and Kroy, Daniela C. and Lazarus, Jeffrey V. and Loomba, Rohit and Manns, Michael P. and Marchesini, Giulio and Nakajima, Atsushi and Negro, Francesco and Petta, Salvatore and Ratziu, Vlad and Romero-Gomez, Manuel and Sanyal, Arun and Schattenberg, J{\"o}rn M. and Tacke, Frank and Tanaka, Junko and Trautwein, Christian and Wei, Lai and Zeuzem, Stefan and Ravazi, Homie}, title = {Modeling NAFLD disease burden in China, France, Germany, Italy, Japan, Spain, United Kingdom, and United States for the period 2016-2030}, series = {Journal of Hepatology}, volume = {69}, journal = {Journal of Hepatology}, doi = {10.1016/j.jhep.2018.05.036}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-227286}, pages = {896-904}, year = {2018}, abstract = {Background \& Aims Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are increasingly a cause of cirrhosis and hepatocellular carcinoma globally. This burden is expected to increase as epidemics of obesity, diabetes and metabolic syndrome continue to grow. The goal of this analysis was to use a Markov model to forecast NAFLD disease burden using currently available data. Methods A model was used to estimate NAFLD and NASH disease progression in eight countries based on data for adult prevalence of obesity and type 2 diabetes mellitus (DM). Published estimates and expert consensus were used to build and validate the model projections. Results If obesity and DM level off in the future, we project a modest growth in total NAFLD cases (0-30\%), between 2016-2030, with the highest growth in China as a result of urbanization and the lowest growth in Japan as a result of a shrinking population. However, at the same time, NASH prevalence will increase 15-56\%, while liver mortality and advanced liver disease will more than double as a result of an aging/increasing population. Conclusions NAFLD and NASH represent a large and growing public health problem and efforts to understand this epidemic and to mitigate the disease burden are needed. If obesity and DM continue to increase at current and historical rates, both NAFLD and NASH prevalence are expected to increase. Since both are reversible, public health campaigns to increase awareness and diagnosis, and to promote diet and exercise can help manage the growth in future disease burden. Lay summary Non-alcoholic fatty liver disease and non-alcoholic steatohepatitis can lead to advanced liver disease. Both conditions are becoming increasingly prevalent as the epidemics of obesity and diabetes continue to increase. A mathematical model was built to understand how the disease burden associated with non-alcoholic fatty liver disease and non-alcoholic steatohepatitis will change over time. Results suggest increasing cases of advanced liver disease and liver-related mortality in the coming years.}, language = {en} } @phdthesis{Bredemeyer2024, author = {Bredemeyer, Cynthia Natascha}, title = {Akademisierung und Professionalisierung der Zahnheilkunde, insbesondere der Zahnchirurgie, in W{\"u}rzburg und Unterfranken im 19. Jahrhundert}, doi = {10.25972/OPUS-36387}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-363878}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die Arbeit befasst sich mit der Akademisierung und Professionalisierung der Zahnheilkunde, insbesondere der Zahnchirurgie, in W{\"u}rzburg und Unterfranken im 19. Jahrhundert. Dies wurde insbesondere anhand des zahnchirurgischen Teils der Lehrchirurgischen Instrumentensammlung der Universit{\"a}t W{\"u}rzburg bzw. des Juliusspitals erforscht. Der zahnchirurgische Teil der Instrumentensammlung war bisher noch nicht erforscht worden und besteht aktuell aus 34+1 Instrumenten, die f{\"u}r diese Arbeit komplett katalogisiert wurden. F{\"u}r die Entwicklung der Instrumente im Verlauf des 19. Jahrhunderts wurde die Provenienz der Teilsammlung ergr{\"u}ndet und diese in den Kontext der Akademisierungsbewegung des 19. Jahrhunderts eingeordnet. Die Forschung wurde anhand der tats{\"a}chlich in der Praxis t{\"a}tigen und nach und nach akademisch ausgebildeten Personen nachvollzogen. Hierzu wurden neben den Instrumenten als Quelle die Adressb{\"u}cher der Stadt W{\"u}rzburg und die Matrikel-, Personal- und Vorlesungsverzeichnisse der Universit{\"a}t W{\"u}rzburg des gesamten 19. Jahrhunderts systematisch durchgearbeitet. Außerdem wurden Lehrb{\"u}cher aus dem nichtakademischen zahnchirurigischen Bereich (Bader) mit denen aus dem sich beginnenden akademischen Bereich analysiert. Anhand dieser Forschungsarbeit konnte dargelegt werden, dass die Zahnchirurgie sich analog zur Chiurgie aus dem handwerklichen Bereich abgekoppelt und nach und nach auf verschiedenen Stufen akademisiert hat. Die Zahnchirurgie hat sich "von unten nach oben" durch das Bestreben nichtakademisch ausgebildeter Menschen akademisiert.}, subject = {Zahnchirurgie}, language = {de} } @phdthesis{Morabbian2024, author = {Morabbian, Jasamin}, title = {Etablierung von Stammzell-Sph{\"a}roiden mit inkorporierten Biokeramik-Partikeln zur F{\"o}rderung der osteogenen Differenzierung}, doi = {10.25972/OPUS-36925}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369256}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {In der vorliegenden Dissertationsarbeit wurden Sph{\"a}roide aus mesenchymalen Stammzellen aus dem Fettgewebe oder dem Knochenmark mittels der Micromold-Methode hergestellt. Den Sph{\"a}roiden wurden entweder Calciumphosphat- oder Calcium-Magnesium-Phosphat-Partikel hinzugef{\"u}gt. Zum einen sollte {\"u}berpr{\"u}ft werden, ob die Zugabe von Partikeln die osteogene Differenzierung der Sph{\"a}roide f{\"o}rdert und somit zur weiteren Entwicklung von k{\"o}rpereigenem Knochenersatzmaterial in der regenerativen Medizin beitr{\"a}gt. Zum anderen sollte festgestellt werden, ob eine der beiden Biokeramiken hinsichtlich der osteogenen Differenzierung {\"u}berlegen ist.}, subject = {Stammzelle}, language = {de} } @article{BugaiQuaresmaFriedeletal.2019, author = {Bugai, Andrii and Quaresma, Alexandre J. C. and Friedel, Caroline C. and Lenasi, Tina and D{\"u}ster, Robert and Sibley, Christopher R. and Fujinaga, Koh and Kukanja, Petra and Hennig, Thomas and Blasius, Melanie and Geyer, Matthias and Ule, Jernej and D{\"o}lken, Lars and Barborič, Matjaž}, title = {P-TEFb Activation by RBM7 Shapes a Pro-survival Transcriptional Response to Genotoxic Stress}, series = {Molecular Cell}, volume = {74}, journal = {Molecular Cell}, doi = {10.1016/j.molcel.2019.01.033}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-221726}, pages = {254-267}, year = {2019}, abstract = {DNA damage response (DDR) involves dramatic transcriptional alterations, the mechanisms of which remain ill defined. Here, we show that following genotoxic stress, the RNA-binding motif protein 7 (RBM7) stimulates RNA polymerase II (Pol II) transcription and promotes cell viability by activating the positive transcription elongation factor b (P-TEFb) via its release from the inhibitory 7SK small nuclear ribonucleoprotein (7SK snRNP). This is mediated by activation of p38MAPK, which triggers enhanced binding of RBM7 with core subunits of 7SK snRNP. In turn, P-TEFb relocates to chromatin to induce transcription of short units, including key DDR genes and multiple classes of non-coding RNAs. Critically, interfering with the axis of RBM7 and P-TEFb provokes cellular hypersensitivity to DNA-damage-inducing agents due to activation of apoptosis. Our work uncovers the importance of stress-dependent stimulation of Pol II pause release, which enables a pro-survival transcriptional response that is crucial for cell fate upon genotoxic insult.}, language = {en} } @phdthesis{Laqua2024, author = {Laqua, Caroline}, title = {Association of myocardial tissue characteristics and functional outcome in biopsy-verified myocarditis assessed by cardiac magnetic resonance imaging}, doi = {10.25972/OPUS-36390}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-363903}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {The relation between LV function and cardiac MRI tissue characteristics in separate myocardial segments and their change over time has yet to be explored in myocarditis. Thus, our research aimed to investigate possible associations between global and regional myocardial T1 and T2 times and peak strain in patients with suspected myocarditis. From 2012 to 2015, 129 patients with clinically suspected myocarditis of the prospective, observational MyoRacer-Trial underwent systematic biventricular EMB at baseline and cardiac MRI at baseline and after three months as a follow-up. We divided the LV myocardium into 17 segments and estimated the segmental myocardial strain using FT. We registered T1 and T2 maps to the cine sequences and transferred the segmentations used for FT to ensure conformity of the myocardial segments. Multi-level multivariable linear mixed effects regression was applied to investigate the relation of segmental myocardial strain to relaxation times and their respective change from baseline to follow-up. We found a significant improvement in myocardial peak strain from baseline to follow-up (p < 0.001; all p-values given for likelihood ratio tests) and significant associations between higher T1 and T2 times and lower segmental myocardial peak strain (p ranging from < 0.001 to 0.049). E.g., regression coefficient (Reg. coef.) for segmental radial peak strain in short axis view (SRPS_SAX) and T1 time: -1.9, 95\% CI (-2.6;-1.2) \%/100 ms, p < 0.001. A decrease in T1 and T2 times from baseline to follow-up was also significantly related to a recovery of segmental peak strains (p ranging from < 0.001 to 0.050). E.g., Reg. coef. for SRPS_SAX per ΔT1: -1.8, 95\% CI (-2.5;-1.0) \%/100 ms, p < 0.001. Moreover, the higher the baseline T1 time, the more substantial the functional recovery from baseline to follow-up (p ranging from 0.004 to 0.042, e.g., for SRPS_SAX: Reg. coef. 1.3, 95\% CI (0.4;2.1) \%/100 ms, p 0.004). We did not find an effect modification by the presence of myocarditis in the EMB (p > 0.1). Our cross-sectional and longitudinal analyses provide evidence of dose-dependent correlations between T1 and T2 relaxation times and myocardial peak strain in patients with clinical presentation of myocarditis, regardless of the EMB result. Thus, assessing strain values and mapping relaxation times helps estimate the functional prognosis in patients with clinically suspected myocarditis.}, subject = {Myokarditis}, language = {en} } @article{BaluapuriHofstetterDudvarskiStankovicetal.2019, author = {Baluapuri, Apoorva and Hofstetter, Julia and Dudvarski Stankovic, Nevenka and Endres, Theresa and Bhandare, Pranjali and Vos, Seychelle Monique and Adhikari, Bikash and Schwarz, Jessica Denise and Narain, Ashwin and Vogt, Markus and Wang, Shuang-Yan and D{\"u}ster, Robert and Jung, Lisa Anna and Vanselow, Jens Thorsten and Wiegering, Armin and Geyer, Matthias and Maric, Hans Michael and Gallant, Peter and Walz, Susanne and Schlosser, Andreas and Cramer, Patrick and Eilers, Martin and Wolf, Elmar}, title = {MYC Recruits SPT5 to RNA Polymerase II to Promote Processive Transcription Elongation}, series = {Molecular Cell}, volume = {74}, journal = {Molecular Cell}, doi = {10.1016/j.molcel.2019.02.031}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-221438}, pages = {674-687}, year = {2019}, abstract = {The MYC oncoprotein binds to promoter-proximal regions of virtually all transcribed genes and enhances RNA polymerase II (Pol II) function, but its precise mode of action is poorly understood. Using mass spectrometry of both MYC and Pol II complexes, we show here that MYC controls the assembly of Pol II with a small set of transcription elongation factors that includes SPT5, a subunit of the elongation factor DSIF. MYC directly binds SPT5, recruits SPT5 to promoters, and enables the CDK7-dependent transfer of SPT5 onto Pol II. Consistent with known functions of SPT5, MYC is required for fast and processive transcription elongation. Intriguingly, the high levels of MYC that are expressed in tumors sequester SPT5 into non-functional complexes, thereby decreasing the expression of growth-suppressive genes. Altogether, these results argue that MYC controls the productive assembly of processive Pol II elongation complexes and provide insight into how oncogenic levels of MYC permit uncontrolled cellular growth.}, language = {en} } @article{McCollGrollJungstetal.2018, author = {McColl, Erin and Groll, J{\"u}rgen and Jungst, Tomasz and Dalton, Paul D.}, title = {Design and fabrication of melt electrowritten tubes using intuitive software}, series = {Materials and Design}, volume = {155}, journal = {Materials and Design}, doi = {10.1016/j.matdes.2018.05.036}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223891}, pages = {46-58}, year = {2018}, abstract = {This study approaches the accurate continuous direct-writing onto a cylindrical collector from a mathematical perspective, taking into account the winding angle, cylinder diameter and length required for the final 3D printed tube. Using an additive manufacturing process termed melt electrowriting (MEW), porous tubes intended for tissue engineering applications are fabricated from medical-grade poly(ε-caprolactone) (PCL), validating the mathematically-derived method. For the fabricated tubes in this study, the pore size, winding angle and printed length can all be planned in advance and manufactured as designed. The physical dimensions of the tubes matched theoretical predictions and mechanical testing performed demonstrated that variations in the tubular morphology have a direct impact on their strength. MEWTubes, the web-based application developed and described here, is a particularly useful tool for planning the complex continuous direct writing path required for MEW onto a rotating, cylindrical build surface.}, language = {en} } @article{ColungaHayworthKressetal.2019, author = {Colunga, Thomas and Hayworth, Miranda and Kreß, Sebastian and Reynolds, David M. and Chen, Luoman and Nazor, Kristopher L. and Baur, Johannes and Singh, Amar M. and Loring, Jeanne F. and Metzger, Marco and Dalton, Stephen}, title = {Human Pluripotent Stem Cell-Derived Multipotent Vascular Progenitors of the Mesothelium Lineage Have Utility in Tissue Engineering and Repair}, series = {Cell Reports}, volume = {26}, journal = {Cell Reports}, doi = {10.1016/j.celrep.2019.02.016}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223217}, pages = {2566-2579}, year = {2019}, abstract = {In this report we describe a human pluripotent stem cell-derived vascular progenitor (MesoT) cell of the mesothelium lineage. MesoT cells are multipotent and generate smooth muscle cells, endothelial cells, and pericytes and self-assemble into vessel-like networks in vitro. MesoT cells transplanted into mechanically damaged neonatal mouse heart migrate into the injured tissue and contribute to nascent coronary vessels in the repair zone. When seeded onto decellularized vascular scaffolds, MesoT cells differentiate into the major vascular lineages and self-assemble into vasculature capable of supporting peripheral blood flow following transplantation. These findings demonstrate in vivo functionality and the potential utility of MesoT cells in vascular engineering applications.}, language = {en} } @article{ChhatbarDetjeGrabskietal.2018, author = {Chhatbar, Chintan and Detje, Claudia N. and Grabski, Elena and Borst, Katharina and Spanier, Julia and Ghita, Luca and Elliott, David A. and Jord{\~a}o, Marta Joana Costa and Mueller, Nora and Sutton, James and Prajeeth, Chittappen K. and Gudi, Viktoria and Klein, Michael A. and Prinz, Marco and Bradke, Frank and Stangel, Martin and Kalinke, Ulrich}, title = {Type I Interferon Receptor Signaling of Neurons and Astrocytes Regulates Microglia Activation during Viral Encephalitis}, series = {Cell Reports}, volume = {25}, journal = {Cell Reports}, doi = {10.1016/j.celrep.2018.09.003}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-222456}, pages = {118-129}, year = {2018}, abstract = {In sterile neuroinflammation, a pathological role is proposed for microglia, whereas in viral encephalitis, their function is not entirely clear. Many viruses exploit the odorant system and enter the CNS via the olfactory bulb (OB). Upon intranasal vesicular stomatitis virus instillation, we show an accumulation of activated microglia and monocytes in the OB. Depletion of microglia during encephalitis results in enhanced virus spread and increased lethality. Activation, proliferation, and accumulation of microglia are regulated by type I IFN receptor signaling of neurons and astrocytes, but not of microglia. Morphological analysis of myeloid cells shows that type I IFN receptor signaling of neurons has a stronger impact on the activation of myeloid cells than of astrocytes. Thus, in the infected CNS, the cross talk among neurons, astrocytes, and microglia is critical for full microglia activation and protection from lethal encephalitis.}, language = {en} } @article{KuschBornscheinLorethetal.2018, author = {Kusch, Valentin and Bornschein, Grit and Loreth, Desiree and Bank, Julia and Jordan, Johannes and Baur, David and Watanabe, Masahiko and Kulik, Akos and Heckmann, Manfred and Eilers, Jens and Schmidt, Hartmut}, title = {Munc13-3 Is Required for the Developmental Localization of Ca2+ Channels to Active Zones and the Nanopositioning of Cav2.1 Near Release Sensors}, series = {Cell Reports}, volume = {22}, journal = {Cell Reports}, doi = {10.1016/j.celrep.2018.02.010}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-233468}, pages = {1965-1973}, year = {2018}, abstract = {Spatial relationships between Cav channels and release sensors at active zones (AZs) are a major determinant of synaptic fidelity. They are regulated developmentally, but the underlying molecular mechanisms are largely unclear. Here, we show that Munc13-3 regulates the density of Cav2.1 and Cav2.2 channels, alters the localization of Cav2.1, and is required for the development of tight, nanodomain coupling at parallel-fiber AZs. We combined EGTA application and Ca2+-channel pharmacology in electrophysiological and two-photon Ca2+ imaging experiments with quantitative freeze-fracture immunoelectron microscopy and mathematical modeling. We found that a normally occurring developmental shift from release being dominated by Ca2+ influx through Cav2.1 and Cav2.2 channels with domain overlap and loose coupling (microdomains) to a nanodomain Cav2.1 to sensor coupling is impaired in Munc13-3-deficient synapses. Thus, at AZs lacking Munc13-3, release remained triggered by Cav2.1 and Cav2.2 microdomains, suggesting a critical role of Munc13-3 in the formation of release sites with calcium channel nanodomains.}, language = {en} } @article{GerberKoenigFendtetal.2019, author = {Gerber, Bertram and K{\"o}nig, Christian and Fendt, Markus and Andreatta, Marta and Romanos, Marcel and Pauli, Paul and Yarali, Ayse}, title = {Timing-dependent valence reversal: a principle of reinforcement processing and its possible implications}, series = {Current Opinion in Behavioral Sciences}, volume = {26}, journal = {Current Opinion in Behavioral Sciences}, doi = {10.1016/j.cobeha.2018.12.001}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232933}, pages = {114-120}, year = {2019}, abstract = {Punishment feels bad, but relief upon its termination feels good. As a consequence of such timing-dependent valence reversal, memories of opposite valence can result from associating stimulus A with, for example, the occurrence of punishment (A-) versus punishment termination (-A): A- training results in aversive memory, but -A training in appetitive memory (corresponding effects exist for reward occurrence and termination). Whereas learning through the occurrence of punishment is well studied, much less is known about learning through its termination. Current research investigates how dopaminergic system function contributes to these processes in Drosophila, rats and humans. We argue that dopamine-related psychopathology may entail distortions in learning through punishment termination, and that this may contribute, for example, to non-suicidal self-injury or post-traumatic stress disorder.}, language = {en} } @phdthesis{Fleissner2024, author = {Fleißner, Janik Frank Hans-Werner}, title = {Die Bedeutung von Oncostatin M f{\"u}r die Lipidhom{\"o}ostase Apoe- und Ldlr-deletierter M{\"a}use}, doi = {10.25972/OPUS-28059}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-280592}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {OSM, ein Vertreter der IL-6-Typ-Zytokine, ist nicht nur f{\"u}r entz{\"u}ndliche, sondern auch f{\"u}r metabolische Prozesse von Bedeutung. Vorarbeiten der Arbeitsgruppe GEIER/HERMANNS und Studien von KOMORI et al. legen protektive Eigenschaften des Zytokins nahe, da M{\"a}use, denen OSMR fehlte, Charakteristika des metabolischen Syndroms aufwiesen. Zur eingehenderen Untersuchung der von OSM vermittelten Wirkung auf den murinen Lipidstoffwechsel wurden zwei f{\"u}r die NAFLD und Atherosklerose anf{\"a}llige Modelle herangezogen und jeweils in Gegenwart und Abwesenheit des Osmr studiert: Weibliche Apoe-/-(Osmr-/-) und Ldlr-/-(Osmr-/-) M{\"a}use wurden {\"u}ber einen Zeitraum von zw{\"o}lf Wochen mit westlicher Di{\"a}t gef{\"u}ttert, w{\"o}chentlich gewogen, am Ende der Di{\"a}t geopfert und geerntet. Wildtypische C57Bl/6-M{\"a}use erfuhren die gleiche Behandlung und dienten als Referenzgruppe. Im Rahmen des Promotionsprojektes wurden Leberfettgehalt, Serumlipidspiegel, Lipoproteinfraktionen und Stuhllipide von Apoe-deletierten M{\"a}usen bestimmt und mit bereits vorhandenen Daten der Ldlr-/-(Osmr-/-) und wildtypischen M{\"a}use in Beziehung gesetzt. Expressionsanalysen von am Lipidstoffwechsel beteiligten Genen in Darm-, Leber- und Fettgewebe trugen dazu bei, OSM-abh{\"a}ngige Regulationen aufzudecken. Ldlr-/- Tiere nahmen unter der Di{\"a}t exzessiv zu, hatten hohe Serumspiegel an Leptin, Gluco-se und Lipiden, eine Lebersteatose und, begleitet von einer Induktion des Vldlr, erh{\"o}hte inflammatorische Marker im visceralen Fettgewebe. Der zus{\"a}tzliche Knockout des Osmr ging mit einer geringeren Vldlr-Expression im Fettgewebe und einer hepatozyt{\"a}ren Induktion von Cyp7a1 einher und resultierte in einem metabolisch g{\"u}nstigeren Ph{\"a}notyp. Apoe-defiziente Tiere unterschieden sich hinsichtlich ihrer Gewichtszunahme nicht von Ldlr-/-Osmr-/- und C57Bl/6-M{\"a}usen. {\"U}berraschenderweise zeigten sich im Serum von Apoe-/-Osmr-/- jedoch gegen{\"u}ber Apoe-/- M{\"a}usen erh{\"o}hte Konzentrationen des Gesamt- und VLDL-Cholesterins, der Triglyceride und freien Fetts{\"a}uren. Obwohl Lebern der Apoe-/-Osmr-/- M{\"a}use geringere Ldlr- und Lrp1-mRNA-Spiegel als die der Apoe-/- M{\"a}use aufwiesen, hatten sie einen h{\"o}heren hepatischen Cholesteringehalt. Bei gesteigerter Cpt1a-Expression fiel der hepatische Tri-glyceridgehalt Apoe-deletierter M{\"a}use geringer aus als in Ldlr-/-(Osmr-/-) und wildtypischen Tieren. Unter Umgehung einer Fettgewebsentz{\"u}ndung pr{\"a}sentierten Apoe-defiziente M{\"a}use Hinweise einer inflammatorischen Lebersch{\"a}digung, die pathogenetisch am ehesten mit einer gest{\"o}rten Cholesterinhom{\"o}ostase in Verbindung zu bringen war. Abh{\"a}ngig vom genetischen Hintergrund des Mausmodells hatte OSM sch{\"u}tzende oder sch{\"a}dliche Effekte auf den Lipidmetabolismus. Die Ergebnisse der vorliegenden Arbeit betonen die entscheidende Bedeutung entz{\"u}ndlicher, von OSM modulierter Prozesse f{\"u}r den Fettstoffwechsel in Leber- und Fettgewebe. Weiterf{\"u}hrende Experimente sind n{\"o}tig, um die den Beobachtungen zugrunde liegenden molekularen Mechanismen zu entschl{\"u}sseln.}, subject = {Apolipoprotein E}, language = {de} } @phdthesis{DasgebNitschke2024, author = {Das [geb. Nitschke], Felix Marcel}, title = {DNA-Methylierung und Genexpression von FKPB5 als Teil des Stresshormonsystems bei von Depressionen und Herzinsuffizienz Betroffenen sowie gesunden Kontrollen}, doi = {10.25972/OPUS-36973}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369730}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {FKBP5 stellt im Stresssystem der HPA-Achse ein zentrales Gen bei der Regulation der Sensitivit{\"a}t des Glukokortikoidrezeptors und somit der Reaktion auf Stress dar. Zur Adaptation an Umwelteinfl{\"u}sse ist es selbst in ein komplexes System von Regulationsmechanismen eingebettet, die unter anderem epigenetische Modifikationen in Form von DNA-Methylierung umfassen. Bisherige Studien legen eine starke Assoziation von FKBP5 zu stressinduzierten psychischen Erkrankungen nahe und weisen auf eine Dysregulation der HPA-Achse als m{\"o}glichen Pathomechanismus hin. F{\"u}r die enge klinische Interaktion von Depression und Herzinsuffizienz sowie eine ebenfalls vermutete Rolle der HPA-Achse in der Pathogenese letzterer, k{\"o}nnte FKBP5 daher ein entscheidendes Bindeglied darstellen. Gleichzeitig bietet die Identifikation einer {\"u}ber FKBP5 ausgedr{\"u}ckten Dysregulation der HPA-Achse einen biologischen Befund, der als Marker f{\"u}r das Ansprechen einer antidepressiven Therapie herangezogen werden k{\"o}nnte. Ziel dieser Arbeit war daher die Untersuchung eines m{\"o}glichen Einflusses regulatorischer Parameter von FKBP5 auf die Herzinsuffizienz sowie eine Pr{\"u}fung dieser als m{\"o}gliche Biomarker f{\"u}r einen Erfolg der antidepressiven Therapie. Dazu wurden Blutproben von ProbandInnen der GEParD- bzw. DaCFail-Studie mit Depression, Herzinsuffizienz sowie gesunde Kontrollen untersucht. Durch Pyrosequenzierung bisulfitkonvertierter DNA erfolgte die Bestimmung der Methylierung regulatorischer CpGs. Die Messung der relativen mRNA-Expression erfolgte durch den Einsatz einer qPCR. In der Auswertung fand sich keine differentielle mRNA-Expression oder Methylierung zwischen den vier Untersuchungsgruppen. Allerdings reagierten depressive PatientInnen verglichen mit der Kontrollgruppe mit einer geringeren Zunahme der mRNA-Expression als Reaktion auf den mDST. Das Therapieansprechen in der Depressionsgruppe wiederum war mit einer niedrigeren Methylierung auf CpG7 sowie einer h{\"o}heren mRNA-Expression zu Therapiebeginn assoziiert. Im Behandlungsverlauf f{\"u}hrte eine Abnahme der mRNA-Expression bei den Respondern zu einer Ann{\"a}herung beider Gruppen. Diese Arbeit konnte keine Hinweise f{\"u}r eine Rolle von FKBP5 in der Pathogenese der Herzinsuffizienz finden. Allerdings zeigten die Befunde zur Regulation des Gens bei Glukokortikoidstimulation eine hohe Konstanz zu vorherigen Ergebnissen. In diesen Kontext reihen sich auch die Ergebnisse f{\"u}r das Therapieansprechen ein, die aufgrund einer Herabregulation der HPA-Achse im Therapieverlauf die Idee einer urs{\"a}chlichen HPA-Dysregulation in der Gruppe der Responder bekr{\"a}ftigen. F{\"u}r sich allein genommen lassen sich mRNA-Expression und Methylierung aufgrund mangelnder Sensitivit{\"a}t und Spezifit{\"a}t nicht als Biomarker f{\"u}r das Therapieansprechen einsetzen. Die bisherigen Befunde best{\"a}rken aber eine m{\"o}gliche Rolle in einer Batterie unterschiedlicher Biomarker auf verschiedenen Ebenen, wie Klinik, Psychometrie und Physiologie.}, subject = {Gen FKBP5}, language = {de} } @phdthesis{Wucherpfennig2024, author = {Wucherpfennig, Sophia}, title = {HTS (high-throughput drug screening) zur Untersuchung der Blut-Hirn-Schranken-Permeabilit{\"a}t in vitro beim zerebral metastasierten Mammakarzinom}, doi = {10.25972/OPUS-36996}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-369964}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die Blut-Hirn-Schranke (BHS) stellt eine selektiv durchl{\"a}ssige Barriere dar, die den Austausch von Stoffen zwischen Blut und ZNS kontrolliert und so neuroprotektiv wirkt. Sie verhindert allerdings nicht nur die Passage toxischer Metaboliten, sondern verwehrt auch vielen therapeutischen Wirkstoffen den Zugang zum Gehirn. Die Forschung an Methoden zum Erreichen h{\"o}herer Arzneimittelkonzentrationen im Gehirn ist deshalb essenziell f{\"u}r die Behandlung zerebraler Erkrankungen wie dem zerebral metastasierten Mammakarzinom. Ziel dieser Arbeit war es deshalb, Wirkstoffe zu identifizieren, die die Permeabilit{\"a}t der BHS erh{\"o}hen. Die Substanzdatenbank LO1208 von Sigma-Aldrich wurde im Rahmen eines HTS auf ihre permeabilit{\"a}tsbeeinflussenden Eigenschaften untersucht. Hierbei konnten 31 Substanzen identifiziert werden, welche die Permeabilit{\"a}t von BLECs um mindestens 50 \% erh{\"o}hen. Aus diesen wurden 4-Amino-1,8-naphthalimid (PARP-Inhibitor) und GW2974 (TKI) f{\"u}r eine genauere Analyse ausgew{\"a}hlt. Als dritter Wirkstoff wurde Ibuilast (Inhibitor der PDE4, des MIF sowie des Toll-like-Rezeptor-4) untersucht, wobei dieser keine signifikante Ver{\"a}nderung der Permeabilit{\"a}t bewirkt. Die Messung des TEERs und der Permeabilit{\"a}t f{\"u}r Fluorescein best{\"a}tigten die Ergebnisse aus dem HTS, welches demnach zuk{\"u}nftig f{\"u}r Permeabilit{\"a}tstests eingesetzt werden kann. Die Zellviabilit{\"a}t wird durch 4 Amino-1,8-naphthalmid nicht beeinflusst. GW2974 und Ibudilast zeigen bei 500 µM einen toxischen Einfluss auf MCF-7-Zellen. BLECs werden durch 100 µM GW2974 gehemmt. Es konnte gezeigt werden, dass die erh{\"o}hte Permeabilit{\"a}t mit einer Ver{\"a}nderung der TJ-Proteinexpression einhergeht. 4-Amino-1,8-naphthalimid senkt die Expression von Occludin auf mRNA- und Proteinebene. GW2974 vermindert zus{\"a}tzlich die Expression von VE-Cadherin, Claudin-5 und ZO-1. Dar{\"u}ber hinaus wurde die Wirkung auf Effluxpumpen untersucht. Die Ergebnisse der mRNA- und Protein-expression weichen voneinander ab, weshalb eine genauere Untersuchung der Translationsvorg{\"a}nge sinnvoll erscheint. Glut-1 wird in GW2974 behandelten Zellen {\"u}berexprimiert, was auf eine erh{\"o}hte Aktivit{\"a}t der BLECs hinweist. GW2974 und 4-Amino-1,8-naphthalimid k{\"o}nnten durch ihre permeabilit{\"a}tssteigernde Wirkung die Ansprechrate einer systemischen Behandlung von PatientInnen mit einem zerebral metastasierten Mammakarzinom erh{\"o}hen und somit ihre Prognose verbessern. Detaillierte Studien zu Kombinationstherapien, den notwendigen Wirkstoff-konzentrationen und eventuellen negativen neurologischen Wirkungen sollten erwogen werden.}, subject = {Blut-Hirn-Schranke}, language = {de} } @phdthesis{Banaschewski2024, author = {Banaschewski, Nora Malaika Marcia Cath{\´e}rine}, title = {Erleichterungslernen bei Jugendlichen mit nicht-suizidalem selbstverletzendem Verhalten}, doi = {10.25972/OPUS-32367}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-323673}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Die Erleichterung von einem k{\"o}rperlichen Schmerzreiz besitzt appetitiven Charakter (Leknes et al., 2008; 2011; Seymour et al., 2005), aktiviert belohnungsassoziierte Hirnstrukturen (Leknes et al., 2011; Leknes \& Brock, 2014; Leknes \& Tracey, 2008; Navratilova \& Porreca, 2014) und f{\"o}rdert durch ihre Konditionierbarkeit als Erleichterungslernen bezeichnete appetitive Lern- und Konditionierungsprozesse (Andreatta et al., 2010, 2012; 2013; 2017; Gerber et al., 2014; Tanimoto et al., 2004; Yarali et al., 2008). Die vorliegende Arbeit best{\"a}tigt das angewandte Versuchsparadigma als valides Modell f{\"u}r Erleichterungslernen im Menschen und zeigt erstmals, dass der appetitive Charakter von Schmerzerleichterung auch in Jugendlichen konditionierbar ist. Erfolgreiches Erleichterungslernen zeigte sich dabei in der untersuchten Stichprobe lediglich auf impliziter, nicht aber auf expliziter, kognitiver Ebene. Dies st{\"u}tzt Thesen und vorherige Forschungsbefunde einer Dualit{\"a}t assoziativen Lernens in ein implizites Lernen, welches vornehmlich subkortikale Strukturen erfordert und ein explizites Lernen, das vorrangig kortikale Strukturen wie den pr{\"a}frontalen Cortex involviert (Andreatta et al., 2010; Strack \& Deutsch, 2004; Williams et al., 2001). Die Beobachtungen einer differenten Furcht- versus Erleichterungs-Extinktion best{\"a}rken die Thesen eines diversen neuronalen Hintergrunds dieser beiden Lernformen (Diegelmann et al., 2013; Gerber et al., 2014; Yarali et al., 2009; Yarali \& Gerber, 2010). Gleichzeitig werfen die Studienergebnisse die Frage auf, ob und inwiefern im Erleichterungslernen von Jugendlichen Unterschiede zu jenem in Erwachsenen bestehen. Die Hypothese einer verst{\"a}rkten Akquisition von Erleichterungslernen bei Jugendlichen mit NSSV im Vergleich zu gesunden Jugendlichen ließ sich in der vorliegenden Studie nicht best{\"a}tigen. Somit liefern die Ergebnisse keinen direkten Hinweis darauf, dass ein verst{\"a}rktes Lernen durch Schmerzerleichterung an der {\"A}tiopathogenese von NSSV beteiligt sein k{\"o}nnte. Die vorliegende Arbeit zeigte vielmehr die Tendenz eines abgeschw{\"a}chten impliziten Erleichterungslernens bei den Jugendlichen mit NSSV. Die tendenziellen Gruppenunterschiede ließen sich nicht hinreichend durch eine differente aktuelle Stimmungslage oder durch eine unterschiedlich starke Auspr{\"a}gung aversiver emotionaler Anspannungen oder momentaner Angstaffekte erkl{\"a}ren. Innerhalb der Gruppe Jugendlicher mit NSSV zeigte sich auch kein Hinweis darauf, dass der Erfolg von Erleichterungslernen vom Schweregrad des NSSV oder von der aktuellen Einnahme von Antidepressiva abh{\"a}ngig sein k{\"o}nnte. Explorative Analysen ergaben, dass Gruppeneffekte in der vorliegenden Studie wom{\"o}glich aufgrund einer statistischen Untersch{\"a}tzung, bedingt durch einen zu geringen Stichprobenumfang, nicht das Signifikanzniveau erreichten und dass Unterschiede im Erleichterungslernen von Jugendlichen mit und ohne NSSV tats{\"a}chlich sogar noch st{\"a}rker ausgepr{\"a}gt sein k{\"o}nnten. Somit sollte die vorliegende Arbeit als Pilotstudie f{\"u}r zuk{\"u}nftige gr{\"o}ßer angelegte Studien zu Erleichterungslernen bei NSSV betrachtet werden. Zuk{\"u}nftige Studien erscheinen insbesondere sinnvoll mit Blick auf die hohe klinische sowie gesellschaftliche Relevanz von NSSV f{\"u}r welches, trotz der hohen Pr{\"a}valenzen und des deutlich erh{\"o}hten Morbidit{\"a}ts- und Mortalit{\"a}tsrisikos, zum aktuellen Zeitpunkt noch keine hinreichenden Erkl{\"a}rungsmodelle bestehen. Die Studie best{\"a}tigte das Vorliegen eines erh{\"o}hten Grades aversiver emotionaler Anspannung in Jugendlichen mit NSSV, welcher zuvor nur an Erwachsenen mit einer BPD untersucht und festgestellt worden war (Niedtfeld et al., 2010; Stiglmayr et al., 2005). Die Abnahme negativer Affekte bei den Jugendlichen mit NSSV im Studienverlauf repliziert die Ergebnisse vorheriger Studien, in denen eine Reduktion selbst-berichteter negativer Affekte durch die Beendigung eines Schmerzreizes beobachtet wurde (Bresin et al., 2010; Bresin \& Gordon, 2013). Damit best{\"a}rken die Studienergebnisse bestehende Erkl{\"a}rungsmodelle f{\"u}r NSSV, welche eine entscheidende Beteiligung der k{\"o}rperlichen Schmerzen und der Schmerzerleichterung bei der Selbstverletzung an der Affektregulation vermuten. Weiterhin wirft die vorliegende Arbeit die Frage auf, welche Rolle eine ver{\"a}nderte Wahrnehmung von Schmerz und Schmerzerleichterung in der {\"A}tiopathogenese von NSSV einnimmt und wie diese sich auf Lernprozesse auswirkt. Insgesamt erbr{\"a}chten weitere Erkenntnisse {\"u}ber den potenziellen Zusammenhang von NSSV und abweichendem Erleichterungslernen ein besseres Verst{\"a}ndnis f{\"u}r Mechanismen der Entstehung und Aufrechterhaltung von NSSV und b{\"o}ten zudem m{\"o}glicherweise Ans{\"a}tze f{\"u}r neue Therapiem{\"o}glichkeiten des St{\"o}rungsbildes.}, subject = {Selbstbesch{\"a}digung}, language = {de} } @article{MuehlemannZdziebloFriedrichetal.2018, author = {M{\"u}hlemann, Markus and Zdzieblo, Daniela and Friedrich, Alexandra and Berger, Constantin and Otto, Christoph and Walles, Heike and Koepsell, Hermann and Metzger, Marco}, title = {Altered pancreatic islet morphology and function in SGLT1 knockout mice on a glucose-deficient, fat-enriched diet}, series = {Molecular Metabolism}, volume = {13}, journal = {Molecular Metabolism}, doi = {10.1016/j.molmet.2018.05.011}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224230}, pages = {67-76}, year = {2018}, abstract = {Objectives Glycemic control by medical treatment represents one therapeutic strategy for diabetic patients. The Na+-d-glucose cotransporter 1 (SGLT1) is currently of high interest in this context. SGLT1 is known to mediate glucose absorption and incretin secretion in the small intestine. Recently, inhibition of SGLT1 function was shown to improve postprandial hyperglycemia. In view of the lately demonstrated SGLT1 expression in pancreatic islets, we investigated if loss of SGLT1 affects islet morphology and function. Methods Effects associated with the loss of SGLT1 on pancreatic islet (cyto) morphology and function were investigated by analyzing islets of a SGLT1 knockout mouse model, that were fed a glucose-deficient, fat-enriched diet (SGLT1-/--GDFE) to circumvent the glucose-galactose malabsorption syndrome. To distinguish diet- and Sglt1-/--dependent effects, wildtype mice on either standard chow (WT-SC) or the glucose-free, fat-enriched diet (WT-GDFE) were used as controls. Feeding a glucose-deficient, fat-enriched diet further required the analysis of intestinal SGLT1 expression and function under diet-conditions. Results Consistent with literature, our data provide evidence that small intestinal SGLT1 mRNA expression and function is regulated by nutrition. In contrast, pancreatic SGLT1 mRNA levels were not affected by the applied diet, suggesting different regulatory mechanisms for SGLT1 in diverse tissues. Morphological changes such as increased islet sizes and cell numbers associated with changes in proliferation and apoptosis and alterations of the β- and α-cell population are specifically observed for pancreatic islets of SGLT1-/--GDFE mice. Glucose stimulation revealed no insulin response in SGLT1-/--GDFE mice while WT-GDFE mice displayed only a minor increase of blood insulin. Irregular glucagon responses were observed for both, SGLT1-/--GDFE and WT-GDFE mice. Further, both animal groups showed a sustained release of GLP-1 compared to WT-SC controls. Conclusion Loss or impairment of SGLT1 results in abnormal pancreatic islet (cyto)morphology and disturbed islet function regarding the insulin or glucagon release capacity from β- or α-cells, respectively. Consequently, our findings propose a new, additional role for SGLT1 maintaining proper islet structure and function.}, language = {en} } @article{HaukeHorvathGrossetal.2018, author = {Hauke, Jan and Horvath, Judit and Groß, Eva and Gehrig, Andrea and Honisch, Ellen and Hackmann, Karl and Schmidt, Gunnar and Arnold, Norbert and Faust, Ulrike and Sutter, Christian and Hentschel, Julia and Wang-Gohrke, Shan and Smogavec, Mateja and Weber, Bernhard H. F. and Weber-Lassalle, Nana and Weber-Lassalle, Konstantin and Borde, Julika and Ernst, Corinna and Altm{\"u}ller, Janine and Volk, Alexander E. and Thiele, Holger and H{\"u}bbel, Verena and N{\"u}rnberg, Peter and Keupp, Katharina and Versmold, Beatrix and Pohl, Esther and Kubisch, Christian and Grill, Sabine and Paul, Victoria and Herold, Natalie and Lichey, Nadine and Rhiem, Kerstin and Ditsch, Nina and Ruckert, Christian and Wappenschmidt, Barbara and Auber, Bernd and Rump, Andreas and Niederacher, Dieter and Haaf, Thomas and Ramser, Juliane and Dworniczak, Bernd and Engel, Christoph and Meindl, Alfons and Schmutzler, Rita K. and Hahnen, Eric}, title = {Gene panel testing of 5589 BRCA1/2-negative index patients with breast cancer in a routine diagnostic setting: results of the German Consortium for Hereditary Breast and Ovarian Cancer}, series = {Cancer Medicine}, journal = {Cancer Medicine}, doi = {10.1002/cam4.1376}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-227902}, pages = {1349-1358}, year = {2018}, abstract = {The prevalence of germ line mutations in non-BRCA1/2 genes associated with hereditary breast cancer (BC) is low, and the role of some of these genes in BC predisposition and pathogenesis is conflicting. In this study, 5589 consecutive BC index patients negative for pathogenic BRCA1/2 mutations and 2189 female controls were screened for germ line mutations in eight cancer predisposition genes (ATM, CDH1, CHEK2, NBN, PALB2, RAD51C, RAD51D, and TP53). All patients met the inclusion criteria of the German Consortium for Hereditary Breast and Ovarian Cancer for germ line testing. The highest mutation prevalence was observed in the CHEK2 gene (2.5\%), followed by ATM (1.5\%) and PALB2 (1.2\%). The mutation prevalence in each of the remaining genes was 0.3\% or lower. Using Exome Aggregation Consortium control data, we confirm significant associations of heterozygous germ line mutations with BC for ATM (OR: 3.63, 95\%CI: 2.67-4.94), CDH1 (OR: 17.04, 95\%CI: 3.54-82), CHEK2 (OR: 2.93, 95\%CI: 2.29-3.75), PALB2 (OR: 9.53, 95\%CI: 6.25-14.51), and TP53 (OR: 7.30, 95\%CI: 1.22-43.68). NBN germ line mutations were not significantly associated with BC risk (OR:1.39, 95\%CI: 0.73-2.64). Due to their low mutation prevalence, the RAD51C and RAD51D genes require further investigation. Compared with control datasets, predicted damaging rare missense variants were significantly more prevalent in CHEK2 and TP53 in BC index patients. Compared with the overall sample, only TP53 mutation carriers show a significantly younger age at first BC diagnosis. We demonstrate a significant association of deleterious variants in the CHEK2, PALB2, and TP53 genes with bilateral BC. Both, ATM and CHEK2, were negatively associated with triple-negative breast cancer (TNBC) and estrogen receptor (ER)-negative tumor phenotypes. A particularly high CHEK2 mutation prevalence (5.2\%) was observed in patients with human epidermal growth factor receptor 2 (HER2)-positive tumors.}, language = {en} } @article{CharbonnierBaradaranSatoetal.2019, author = {Charbonnier, Baptiste and Baradaran, Aslan and Sato, Daisuke and Alghamdi, Osama and Zhang, Zishuai and Zhang, Yu-Ling and Gbureck, Uwe and Gilardino, Mirko and Harvey, Edward and Makhoul, Nicholas and Barralet, Jake}, title = {Material-Induced Venosome-Supported Bone Tubes}, series = {Advanced Science}, volume = {6}, journal = {Advanced Science}, doi = {10.1002/advs.201900844}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-222318}, year = {2019}, abstract = {The development of alternatives to vascular bone grafts, the current clinical standard for the surgical repair of large segmental bone defects still today represents an unmet medical need. The subcutaneous formation of transplantable bone has been successfully achieved in scaffolds axially perfused by an arteriovenous loop (AVL) and seeded with bone marrow stromal cells or loaded with inductive proteins. Although demonstrating clinical potential, AVL-based approaches involve complex microsurgical techniques and thus are not in widespread use. In this study, 3D-printed microporous bioceramics, loaded with autologous total bone marrow obtained by needle aspiration, are placed around and next to an unoperated femoral vein for 8 weeks to assess the effect of a central flow-through vein on bone formation from marrow in a subcutaneous site. A greater volume of new bone tissue is observed in scaffolds perfused by a central vein compared with the nonperfused negative control. These analyses are confirmed and supplemented by calcified and decalcified histology. This is highly significant as it indicates that transplantable vascularized bone can be grown using dispensable vein and marrow tissue only. This is the first report illustrating the capacity of an intrinsic vascularization by a single vein to support ectopic bone formation from untreated marrow.}, language = {en} }