TY - JOUR A1 - Robinson, Thomas M. A1 - Hutmacher, Dietmar W. A1 - Dalton, Paul D. T1 - The next frontier in melt electrospinning: taming the jet JF - Advanced Functional Materials N2 - There is a specialized niche for the electrohydrodynamic jetting of melts, from biomedical products to filtration and soft matter applications. The next frontier includes optics, microfluidics, flexible electronic devices, and soft network composites in biomaterial science and soft robotics. The recent emphasis on reproducibly direct‐writing continual molten jets has enabled a spectrum of contemporary microscale 3D objects to be fabricated. One strong suit of melt processing is the capacity for the jet to solidify rapidly into a fiber, thus fixing a particular structure into position. The ability to direct‐write complex and multiscaled architectures and structures has greatly contributed to a large number of recent studies, explicitly, toward fiber–hydrogel composites and fugitive inks, and has expanded into several biomedical applications such as cartilage, skin, periosteum, and cardiovascular tissue engineering. Following the footsteps of a publication that summarized melt electrowriting literature up to 2015, the most recent literature from then until now is reviewed to provide a continuous and comprehensive timeline that demonstrates the latest advances as well as new perspectives for this emerging technology. KW - 3D printing KW - additive manufacturing KW - eletrhydrodynamic KW - melt electrospinning writing Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-204819 VL - 29 ER - TY - JOUR A1 - Stauss, Dennis A1 - Brunner, Cornelia A1 - Berberich-Siebelt, Friederike A1 - Höpken, Uta E. A1 - Lipp, Martin A1 - Müller, Gerd T1 - The transcriptional coactivator Bob1 promotes the development of follicular T helper cells via Bcl6 JF - Embo Journal N2 - Follicular T helper (Tfh) cells are key regulators of the germinal center reaction and long-term humoral immunity. Tfh cell differentiation requires the sustained expression of the transcriptional repressor Bcl6; however, its regulation in CD4\(^+\) T cells is incompletely understood. Here, we report that the transcriptional coactivator Bob1, encoded by the Pou2af1 gene, promotes Bcl6 expression and Tfh cell development. We found that Bob1 together with the octamer transcription factors Oct1/Oct2 can directly bind to and transactivate the Bcl6 and Btla promoters. Mixed bone marrow chimeras revealed that Bob1 is required for the expression of normal levels of Bcl6 and BTLA, thereby controlling the pool size and composition of the Tfh compartment in a T cell-intrinsic manner. Our data indicate that T cell-expressed Bob1 is directly involved in Tfh cell differentiation and required for mounting normal T cell-dependent B-cell responses. KW - follicular T helper cells KW - germinal center KW - humoral immunity KW - Pou2af1 KW - T cell differentiation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189506 VL - 35 IS - 8 ER - TY - JOUR A1 - Beck, Hanna A1 - Titze, Stephanie I. A1 - Hübner, Silvia A1 - Busch, Martin A1 - Schlieper, Georg A1 - Schultheiss, Ulla T. A1 - Wanner, Christoph A1 - Kronenberg, Florian A1 - Krane, Vera A1 - Eckardt, Kai-Uwe A1 - Köttgen, Anna T1 - Heart Failure in a Cohort of Patients with Chronic Kidney Disease: The GCKD Study JF - PLoS ONE N2 - Background and Aims Chronic kidney disease (CKD) is a risk factor for development and progression of heart failure (HF). CKD and HF share common risk factors, but few data exist on the prevalence, signs and symptoms as well as correlates of HF in populations with CKD of moderate severity. We therefore aimed to examine the prevalence and correlates of HF in the German Chronic Kidney Disease (GCKD) study, a large observational prospective study. Methods and Results We analyzed data from 5,015 GCKD patients aged 18-74 years with an estimated glomerular filtration rate (eGFR) of <60 ml/min/1.73m\(^{2}\) or with an eGFR >= 60 and overt proteinuria (>500 mg/d). We evaluated a definition of HF based on the Gothenburg score, a clinical HF score used in epidemiological studies (Gothenburg HF), and self-reported HF. Factors associated with HF were identified using multivariable adjusted logistic regression. The prevalence of Gothenburg HF was 43% (ranging from 24% in those with eGFR >90 to 59% in those with eGFR<30 ml/min/1.73m2). The corresponding estimate for self-reported HF was 18% (range 5%-24%). Lower eGFR was significantly and independently associated with the Gothenburg definition of HF (p-trend <0.001). Additional significantly associated correlates included older age, female gender, higher BMI, hypertension, diabetes mellitus, valvular heart disease, anemia, sleep apnea, and lower educational status. Conclusions The burden of self-reported and Gothenburg HF among patients with CKD is high. The proportion of patients who meet the criteria for Gothenburg HF in a European cohort of patients with moderate CKD is more than twice as high as the prevalence of self-reported HF. However, because of the shared signs, symptoms and medications of HF and CKD, the Gothenburg score cannot be used to reliably define HF in CKD patients. Our results emphasize the need for early screening for HF in patients with CKD. KW - global outcomes KW - cardiovascularm disease KW - consensus conference KW - men born KW - insufficiency KW - epidemiology KW - European Society KW - atherosclerosis risk KW - United States KW - glomerular filtration rate KW - KDIGO Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143315 VL - 10 IS - 4 ER - TY - JOUR A1 - Janssen, Jan P. A1 - Hoffmann, Jan V. A1 - Kanno, Takayuki A1 - Nose, Naoko A1 - Grunz, Jan-Peter A1 - Onoguchi, Masahisa A1 - Chen, Xinyu A1 - Lapa, Constantin A1 - Buck, Andreas K. A1 - Higuchi, Takahiro T1 - Capabilities of multi-pinhole SPECT with two stationary detectors for in vivo rat imaging JF - Scientific Reports N2 - We aimed to investigate the image quality of the U-SPECT5/CT E-Class a micro single-photon emission computed tomography (SPECT) system with two large stationary detectors for visualization of rat hearts and bones using clinically available \(^{99m}\)Tc-labelled tracers. Sensitivity, spatial resolution, uniformity and contrast-to-noise ratio (CNR) of the small-animal SPECT scanner were investigated in phantom studies using an ultra-high-resolution rat and mouse multi-pinhole collimator (UHR-RM). Point source, hot-rod, and uniform phantoms with \(^{99m}\)Tc-solution were scanned for high-count performance assessment and count levels equal to animal scans, respectively. Reconstruction was performed using the similarity-regulated ordered-subsets expectation maximization (SROSEM) algorithm with Gaussian smoothing. Rats were injected with similar to 100 MBq [\(^{99m}\)TcTc-MIBI or similar to 150 MBq [\(^{99m}\)Tc]Tc-HMDP and received multi-frame micro-SPECT imaging after tracer distribution. Animal scans were reconstructed for three different acquisition times and post-processed with different sized Gaussian filters. Following reconstruction, CNR was calculated and image quality evaluated by three independent readers on a five-point scale from 1="very poor" to 5="very good". Point source sensitivity was 567 cps/MBq and radioactive rods as small as 1.2 mm were resolved with the UHR-RM collimator. Collimator-dependent uniformity was 55.5%. Phantom CNR improved with increasing rod size, filter size and activity concentration. Left ventricle and bone structures were successfully visualized in rat experiments. Image quality was strongly affected by the extent of post-filtering, whereas scan time did not have substantial influence on visual assessment. Good image quality was achieved for resolution range greater than 1.8 mm in bone and 2.8 mm in heart. The recently introduced small animal SPECT system with two stationary detectors and UHR-RM collimator is capable to provide excellent image quality in heart and bone scans in a rat using standardized reconstruction parameters and appropriate post-filtering. However, there are still challenges in achieving maximum system resolution in the sub-millimeter range with in vivo settings under limited injection dose and acquisition time. KW - small animal SPECT KW - HMDP hydroxymethylene diphosphonate KW - skeletal KW - quality KW - scanner Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230616 VL - 10 ER - TY - JOUR A1 - Deeb, Wissam A1 - Giordano, James J. A1 - Rossi, Peter J. A1 - Mogilner, Alon Y. A1 - Gunduz, Aysegul A1 - Judy, Jack W. A1 - Klassen, Bryan T. A1 - Butson, Christopher R. A1 - Van Horne, Craig A1 - Deny, Damiaan A1 - Dougherty, Darin D. A1 - Rowell, David A1 - Gerhardt, Greg A. A1 - Smith, Gwenn S. A1 - Ponce, Francisco A. A1 - Walker, Harrison C. A1 - Bronte-Stewart, Helen M. A1 - Mayberg, Helen S. A1 - Chizeck, Howard J. A1 - Langevin, Jean-Philippe A1 - Volkmann, Jens A1 - Ostrem, Jill L. A1 - Shute, Jonathan B. A1 - Jimenez-Shahed, Joohi A1 - Foote, Kelly D. A1 - Wagle Shukla, Aparna A1 - Rossi, Marvin A. A1 - Oh, Michael A1 - Pourfar, Michael A1 - Rosenberg, Paul B. A1 - Silburn, Peter A. A1 - de Hemptine, Coralie A1 - Starr, Philip A. A1 - Denison, Timothy A1 - Akbar, Umer A1 - Grill, Warren M. A1 - Okun, Michael S. T1 - Proceedings of the Fourth Annual Deep Brain Stimulation Think Tank: A Review of Emerging Issues and Technologies JF - Frontiers in Integrative Neuroscience N2 - This paper provides an overview of current progress in the technological advances and the use of deep brain stimulation (DBS) to treat neurological and neuropsychiatric disorders, as presented by participants of the Fourth Annual DBS Think Tank, which was convened in March 2016 in conjunction with the Center for Movement Disorders and Neurorestoration at the University of Florida, Gainesveille FL, USA. The Think Tank discussions first focused on policy and advocacy in DBS research and clinical practice, formation of registries, and issues involving the use of DBS in the treatment of Tourette Syndrome. Next, advances in the use of neuroimaging and electrochemical markers to enhance DBS specificity were addressed. Updates on ongoing use and developments of DBS for the treatment of Parkinson's disease, essential tremor, Alzheimer's disease, depression, post-traumatic stress disorder, obesity, addiction were presented, and progress toward innovation(s) in closed-loop applications were discussed. Each section of these proceedings provides updates and highlights of new information as presented at this year's international Think Tank, with a view toward current and near future advancement of the field. KW - deep brain stimulation KW - Parkinson’s disease KW - Alzheimer’s disease KW - closed-loop KW - depression KW - post-traumatic stress disorder KW - Tourette syndrome KW - DARPA Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-168493 VL - 10 IS - 38 ER - TY - JOUR A1 - Dresen, Ellen A1 - Pimiento, Jose M. A1 - Patel, Jayshil J. A1 - Heyland, Daren K. A1 - Rice, Todd W. A1 - Stoppe, Christian T1 - Overview of oxidative stress and the role of micronutrients in critical illness JF - Journal of Parenteral and Enteral Nutrition N2 - Inflammation and oxidative stress represent physiological response mechanisms to different types of stimuli and injury during critical illness. Its proper regulation is fundamental to cellular and organismal survival and are paramount to outcomes and recovery from critical illness. A proper maintenance of the delicate balance between inflammation, oxidative stress, and immune response is crucial for resolution from critical illness with important implications for patient outcome. The extent of inflammation and oxidative stress under normal conditions is limited by the antioxidant defense system of the human body, whereas the antioxidant capacity is commonly significantly compromised, and serum levels of micronutrients and vitamins significantly depleted in patients who are critically ill. Hence, the provision of antioxidants and anti-inflammatory nutrients may help to reduce the extent of oxidative stress and therefore improve clinical outcomes in patients who are critically ill. As existing evidence of the beneficial effects of antioxidant supplementation in patients who are critically ill is still unclear, actual findings about the most promising anti-inflammatory and antioxidative candidates selenium, vitamin C, zinc, and vitamin D will be discussed in this narrative review. The existing evidence provided so far demonstrates that several factors need to be considered to determine the efficacy of an antioxidant supplementation strategy in patients who are critically ill and indicates the need for adequately designed multicenter prospective randomized control trials to evaluate the clinical significance of different types and doses of micronutrients and vitamins in selected groups of patients with different types of critical illness. KW - critical illness KW - vitamins KW - vitamin C KW - inflammation KW - medical nutrition therapy KW - oxidative stress KW - selenium KW - trace elements KW - micronutrients KW - vitamin D KW - zinc Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-318186 VL - 47 SP - S38 EP - S49 ER - TY - JOUR A1 - Kobsar, Anna A1 - Koehnlechner, Karina A1 - Klingler, Philipp A1 - Niklaus, Marius A1 - Zeller-Hahn, Julia A1 - Koessler, Angela A1 - Weber, Katja A1 - Boeck, Markus A1 - Koessler, Juergen T1 - The effect of short-term refrigeration on platelet responsiveness JF - Scientific Reports N2 - Storage of platelet concentrates (PC) at cold temperature (CT) is discussed as an alternative to the current standard of storage at room temperature (RT). Recently, we could show that cold-induced attenuation of inhibitory signaling is an important mechanism promoting platelet reactivity. For developing strategies in blood banking, it is required to elucidate the time-dependent onset of facilitated platelet activation. Thus, freshly prepared platelet-rich-plasma (PRP) was stored for 1 and 2 h at CT (2–6 °C) or at RT (20–24 °C), followed by subsequent comparative analysis. Compared to RT, basal and induced vasodilator-stimulated phosphoprotein (VASP) phosphorylation levels were decreased under CT within 1 h by approximately 20%, determined by Western blot analysis and flow cytometry. Concomitantly, ADP- and collagen-induced threshold aggregation values were enhanced by up to 30–40%. Furthermore, platelet-covered areas on collagen-coated slides and aggregate formation under flow conditions were increased after storage at CT, in addition to induced activation markers. In conclusion, a time period of 1–2 h for refrigeration is sufficient to induce an attenuation of inhibitory signaling, accompanied with an enhancement of platelet responsiveness. Short-term refrigeration may be considered as a rational approach to obtain PC with higher functional reactivity for the treatment of hemorrhage. KW - short‑term refrigeration KW - platelet responsiveness KW - cold temperature Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-301390 VL - 12 IS - 1 ER - TY - JOUR A1 - Page, Lukas A1 - Wallstabe, Julia A1 - Lother, Jasmin A1 - Bauser, Maximilian A1 - Kniemeyer, Olaf A1 - Strobel, Lea A1 - Voltersen, Vera A1 - Teutschbein, Janka A1 - Hortschansky, Peter A1 - Morton, Charles Oliver A1 - Brakhage, Axel A. A1 - Topp, Max A1 - Einsele, Hermann A1 - Wurster, Sebastian A1 - Loeffler, Juergen T1 - CcpA- and Shm2-Pulsed Myeloid Dendritic Cells Induce T-Cell Activation and Enhance the Neutrophilic Oxidative Burst Response to Aspergillus fumigatus JF - Frontiers in Immunology N2 - Aspergillus fumigatus causes life-threatening opportunistic infections in immunocompromised patients. As therapeutic outcomes of invasive aspergillosis (IA) are often unsatisfactory, the development of targeted immunotherapy remains an important goal. Linking the innate and adaptive immune system, dendritic cells are pivotal in anti-Aspergillus defense and have generated interest as a potential immunotherapeutic approach in IA. While monocyte-derived dendritic cells (moDCs) require ex vivo differentiation, antigen-pulsed primary myeloid dendritic cells (mDCs) may present a more immediate platform for immunotherapy. To that end, we compared the response patterns and cellular interactions of human primary mDCs and moDCs pulsed with an A. fumigatus lysate and two A. fumigatus proteins (CcpA and Shm2) in a serum-free, GMP-compliant medium. CcpA and Shm2 triggered significant upregulation of maturation markers in mDCs and, to a lesser extent, moDCs. Furthermore, both A. fumigatus proteins elicited the release of an array of key pro-inflammatory cytokines including TNF-α, IL-1β, IL-6, IL-8, and CCL3 from both DC populations. Compared to moDCs, CcpA- and Shm2-pulsed mDCs exhibited greater expression of MHC class II antigens and stimulated stronger proliferation and IFN-γ secretion from autologous CD4\(^+\) and CD8\(^+\) T-cells. Moreover, supernatants of CcpA- and Shm2-pulsed mDCs significantly enhanced the oxidative burst in allogeneic neutrophils co-cultured with A. fumigatus germ tubes. Taken together, our in vitro data suggest that ex vivo CcpA- and Shm2-pulsed primary mDCs have the potential to be developed into an immunotherapeutic approach to tackle IA. KW - antigens KW - dendritic cells KW - cytokines KW - host defense KW - immunotherapy KW - Aspergillus Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239493 SN - 1664-3224 VL - 12 ER - TY - JOUR A1 - Claus, Heike A1 - Hubert, Kerstin A1 - Becher, Dörte A1 - Otto, Andreas A1 - Pawlik, Marie-Christin A1 - Lappann, Ines A1 - Strobel, Lea A1 - Vogel, Ulrich A1 - Johswich, Kay T1 - A homopolymeric adenosine tract in the promoter region of nspA influences factor H-mediated serum resistance in Neisseria meningitidis JF - Scientific Reports N2 - Although usually asymptomatically colonizing the human nasopharynx, the Gram-negative bacterium Neisseria meningitidis (meningococcus) can spread to the blood stream and cause invasive disease. For survival in blood, N. meningitidis evades the complement system by expression of a polysaccharide capsule and surface proteins sequestering the complement regulator factor H (fH). Meningococcal strains belonging to the sequence type (ST-) 41/44 clonal complex (cc41/44) cause a major proportion of serogroup B meningococcal disease worldwide, but they are also common in asymptomatic carriers. Proteome analysis comparing cc41/44 isolates from invasive disease versus carriage revealed differential expression levels of the outer membrane protein NspA, which binds fH. Deletion of nspA reduced serum resistance and NspA expression correlated with fH sequestration. Expression levels of NspA depended on the length of a homopolymeric tract in the nspA promoter: A 5-adenosine tract dictated low NspA expression, whereas a 6-adenosine motif guided high NspA expression. Screening German cc41/44 strain collections revealed the 6-adenosine motif in 39% of disease isolates, but only in 3.4% of carriage isolates. Thus, high NspA expression is associated with disease, but not strictly required. The 6-adenosine nspA promoter is most common to the cc41/44, but is also found in other hypervirulent clonal complexes. KW - Meningitis KW - Pathogens Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-200956 VL - 9 ER - TY - JOUR A1 - Bankoglu, Ezgi Eyluel A1 - Arnold, Charlotte A1 - Hering, Ilona A1 - Hankir, Mohammed A1 - Seyfried, Florian A1 - Stopper, Helga T1 - Decreased chromosomal damage in lymphocytes of obese patients after bariatric surgery JF - Scientific Reports N2 - The number of bariatric surgeries being performed worldwide has markedly risen. While the improvement in obesity-associated comorbidities after bariatric surgery is well-established, very little is known about its impact on cancer risk. The peripheral lymphocyte micronucleus test is a widely used method for the monitoring of chromosomal damage levels in vivo, and micronucleus frequency positively correlates with cancer risk. Therefore, the aim of this study was to compare the micronucleus frequency before and after bariatric surgery in obese subjects. Peripheral blood mononuclear cells were collected from 45 obese subjects before and at two time-points after bariatric surgery (6 and 12 months) to assess spontaneous micronucleus frequency. Consistent with the increased cancer risk previously shown, bariatric surgery-induced weight loss led to a significant reduction in lymphocyte micronucleus frequency after 12 months. Interestingly, comorbidities such as type 2 diabetes mellitus and metabolic syndrome further seemed to have an impact on the lymphocyte micronucleus frequency. Our findings may indicate a successful reduction of cancer risk in patients following weight loss caused by bariatric surgery. KW - obesity KW - bariatric surgery KW - cancer risk Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-177090 VL - 8 IS - 11195 ER - TY - JOUR A1 - Friedrich, Maximilian A1 - Hartig, Johannes A1 - Prüss, Harald A1 - Ip, Wang Chi A1 - Volkmann, Jens T1 - Rapidly progressive dementia: Extending the spectrum of GFAP-astrocytopathies? JF - Annals of Clinical and Translational Neurology N2 - Autoimmune glial fibrillary acidic protein astrocytopathy (GFAP-A) is a steroid-responsive meningoencephalomyelitis, sometimes presenting with atypical clinical signs such as movement disorders or psychiatric and autonomic features. Beyond clinical presentation and imaging, diagnosis relies on detection of GFAP-antibodies (AB) in CSF. Using quantitative behavioral, serologic, and immunohistochemical analyses, we characterize two patients longitudinally over 18–24 months who presented with rapidly progressive neurocognitive deterioration in the context of GFAP-AB in CSF and unremarkable cranial MRI studies. Intensified immunotherapy was associated with clinical stabilization. The value of GFAP-AB screening in selected cases of rapidly progressive dementias is discussed. KW - GFAP-astrocytopathies KW - dementia Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-312957 VL - 9 IS - 3 ER - TY - JOUR A1 - Manukjan, Georgi A1 - Wiegering, Verena A1 - Reindl, Tobias A1 - Strauß, Gabriele A1 - Klopocki, Eva A1 - Schulze, Harald A1 - Andres, Oliver T1 - Novel variants in FERMT3 and RASGRP2 - Genetic linkage in Glanzmann-like bleeding disorders JF - Pediatric Blood & Cancer N2 - Defects of platelet intracellular signaling can result in severe platelet dysfunction. Several mutations in each of the linked genes FERMT3 and RASGRP2 on chromosome 11 causing a Glanzmann‐like bleeding phenotype have been identified so far. We report on novel variants in two unrelated pediatric patients with severe bleeding diathesis—one with leukocyte adhesion deficiency type III due to a homozygous frameshift in FERMT3 and the other with homozygous variants in both, FERMT3 and RASGRP2 . We focus on the challenging genetic and functional variant assessment and aim to accentuate the risk of obtaining misleading results due to the phenomenon of genetic linkage. KW - bleding disorders other than hemophilia KW - hematology KW - hemostasis and thrombosis KW - platelet disorders Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-208129 VL - 67 IS - 2 ER - TY - JOUR A1 - Tan, Aaron A1 - Babak, Maria V. A1 - Venkatesan, Gopalakrishnan A1 - Lim, Clarissa A1 - Klotz, Karl-Norbert A1 - Herr, Deron Raymond A1 - Cheong, Siew Lee A1 - Federico, Stephanie A1 - Spalluto, Giampiero A1 - Ong, Wei-Yi A1 - Chen, Yu Zong A1 - Loo, Jason Siau Ee A1 - Pastorin, Giorgia T1 - Design, Synthesis and Evaluation of New Indolylpyrimidylpiperazines for Gastrointestinal Cancer Therapy JF - Molecules N2 - Human A3 adenosine receptor hA3AR has been implicated in gastrointestinal cancer, where its cellular expression has been found increased, thus suggesting its potential as a molecular target for novel anticancer compounds. Observation made in our previous work indicated the importance of the carbonyl group of amide in the indolylpyrimidylpiperazine (IPP) for its human A2A adenosine receptor (hA2AAR) subtype binding selectivity over the other AR subtypes. Taking this observation into account, we structurally modified an indolylpyrimidylpiperazine (IPP) scaffold, 1 (a non-selective adenosine receptors’ ligand) into a modified IPP (mIPP) scaffold by switching the position of the carbonyl group, resulting in the formation of both ketone and tertiary amine groups in the new scaffold. Results showed that such modification diminished the A2A activity and instead conferred hA3AR agonistic activity. Among the new mIPP derivatives (3–6), compound 4 showed potential as a hA3AR partial agonist, with an Emax of 30% and EC50 of 2.89 ± 0.55 μM. In the cytotoxicity assays, compound 4 also exhibited higher cytotoxicity against both colorectal and liver cancer cells as compared to normal cells. Overall, this new series of compounds provide a promising starting point for further development of potent and selective hA3AR partial agonists for the treatment of gastrointestinal cancers. KW - gastrointestinal cancer KW - hA3AR KW - partial agonists KW - indolylpyrimidylpiperazines Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-193271 SN - 1420-3049 VL - 24 IS - 20 ER - TY - JOUR A1 - Kreß, Julia Katharina Charlotte A1 - Jessen, Christina A1 - Marquardt, André A1 - Hufnagel, Anita A1 - Meierjohann, Svenja T1 - NRF2 enables EGFR signaling in melanoma cells JF - International Journal of Molecular Sciences N2 - Receptor tyrosine kinases (RTK) are rarely mutated in cutaneous melanoma, but the expression and activation of several RTK family members are associated with a proinvasive phenotype and therapy resistance. Epidermal growth factor receptor (EGFR) is a member of the RTK family and is only expressed in a subgroup of melanomas with poor prognosis. The insight into regulators of EGFR expression and activation is important for the understanding of the development of this malignant melanoma phenotype. Here, we describe that the transcription factor NRF2, the master regulator of the oxidative and electrophilic stress response, mediates the expression and activation of EGFR in melanoma by elevating the levels of EGFR as well as its ligands EGF and TGFα. ChIP sequencing data show that NRF2 directly binds to the promoter of EGF, which contains a canonical antioxidant response element. Accordingly, EGF is induced by oxidative stress and is also increased in lung adenocarcinoma and head and neck carcinoma with mutationally activated NRF2. In contrast, regulation of EGFR and TGFA occurs by an indirect mechanism, which is enabled by the ability of NRF2 to block the activity of the melanocytic lineage factor MITF in melanoma. MITF effectively suppresses EGFR and TGFA expression and therefore serves as link between NRF2 and EGFR. As EGFR was previously described to stimulate NRF2 activity, the mutual activation of NRF2 and EGFR pathways was investigated. The presence of NRF2 was necessary for full EGFR pathway activation, as NRF2-knockout cells showed reduced AKT activation in response to EGF stimulation compared to controls. Conversely, EGF led to the nuclear localization and activation of NRF2, thereby demonstrating that NRF2 and EGFR are connected in a positive feedback loop in melanoma. In summary, our data show that the EGFR-positive melanoma phenotype is strongly supported by NRF2, thus revealing a novel maintenance mechanism for this clinically challenging melanoma subpopulation. KW - EGFR KW - NRF2 KW - NFE2L2 KW - KEAP1 KW - MITF-low KW - TGF-alpha KW - EGF KW - NSCLC KW - HNSC Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260222 SN - 1422-0067 VL - 22 IS - 8 ER - TY - JOUR A1 - Braun, Alexandra A1 - Evdokimov, Dimitar A1 - Frank, Johanna A1 - Pauli, Paul A1 - Wabel, Thomas A1 - Üçeyler, Nurcan A1 - Sommer, Claudia T1 - Relevance of Religiosity for Coping Strategies and Disability in Patients with Fibromyalgia Syndrome JF - Journal of Religion and Health N2 - Coping strategies are essential for the outcome of chronic pain. This study evaluated religiosity in a cohort of patients with fibromyalgia syndrome (FMS), its effect on pain and other symptoms, on coping and FMS-related disability. A total of 102 FMS patients were recruited who filled in questionnaires, a subgroup of 42 patients participated in a face-to-face interview, and data were evaluated by correlation and regression analyses. Few patients were traditionally religious, but the majority believed in a higher existence and described their spirituality as "transcendence conviction". The coping strategy "praying-hoping" and the ASP dimension "religious orientation" (r = 0.5, P < 0.05) showed a significant relationship independent of the grade of religiosity (P < 0.05). A high grade of belief in a higher existence was negatively associated with the choice of ignoring as coping strategy (r = - 0.4, P < 0.05). Mood and affect-related variables had the highest impact on disability (b = 0.5, P < 0.05). In this cohort, the grade of religiosity played a role in the choice of coping strategies, but had no effects on health and mood outcome. KW - Fibromyalgia syndrome KW - religiosity KW - coping KW - disability Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-269135 SN - 1573-6571 VL - 61 IS - 1 ER - TY - JOUR A1 - Hanke, Kathrin A1 - Rausch, Tanja K. A1 - Sosnowski, Runa A1 - Paul, Pia A1 - Spiegler, Juliane A1 - Müller, Mirja A1 - König, Inke R. A1 - Göpel, Wolfgang A1 - Herting, Egbert A1 - Härtel, Christoph T1 - Early skin-to-skin contact does not affect cerebral tissue oxygenation in preterm infants <32 weeks of gestation JF - Children N2 - Aim: It was the aim of our study to determine the regional cerebral tissue oxygenation saturation (rcSO\(_2\)) as an additional monitoring parameter during early skin-to-skin contact (SSC) in preterm infants with a gestational age of <32 gestational weeks. Methods: We conducted two observational convenience sample studies using additional monitoring with near-infrared spectroscopy (NIRS) in the first 120 h of life: (a) NIRS 1 (gestational age of 26 0/7 to 31 6/7 weeks) and (b) NIRS 2 (gestational age of 24 0/7 to 28 6/7 weeks). The rcSO\(_2\) values were compared between resting time in the incubator (period I), SSC (period II) and handling nursing care (period III). For the comparison, we separated the sequential effects by including a “wash-out phase” of 1 h between each period. Results: During the first 120 h of life 38/53 infants in NIRS 1 and 15/23 infants in NIRS 2 received SSC, respectively. We found no remarkable differences for rcSO\(_2\) values of NIRS 1 patients between SSC time and period I (95% confidence interval (CI) for the difference in %: SSC vs. period I [1; 3]). In NIRS 2, rcSO\(_2\) values during SSC were only 2% lower compared with period I [median [1. quartile; 3. quartile] in %; 78 [73; 82] vs. 80 [74; 85]] but were similar to period III [78 [72; 83]]. In a combined analysis, a small difference in rcSO\(_2\) values between SSC and resting times was found using a generalized linear mixed model that included gender and gestational age (OR 95% CI; 1.178 [1.103; 1.253], p < 0.0001). Episodes below the cut-off for “hypoxia”; e.g., <55%, were comparable during SSC and periods I and III (0.3–2.1%). No FiO\(_2\) adjustment was required in the vast majority of SSC episodes. Conclusions: Our observational data indicate that rcSO\(_2\) values of infants during SSC were comparable to rcSO\(_2\) values during incubator care and resting time. This additional monitoring supports a safe implementation of early SSC in extremely preterm infants in NICUs. KW - regional cerebral oxygenation saturation KW - near infrared spectroscopy KW - skin-to-skin contact KW - preterm infants Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-262290 SN - 2227-9067 VL - 9 IS - 2 ER - TY - JOUR A1 - Gerova, Milan A1 - Wicke, Laura A1 - Chihara, Kotaro A1 - Schneider, Cornelius A1 - Lavigne, Rob A1 - Vogel, Jörg T1 - A grad-seq view of RNA and protein complexes in Pseudomonas aeruginosa under standard and bacteriophage predation conditions JF - mbio N2 - The Gram-negative rod-shaped bacterium Pseudomonas aeruginosa is not only a major cause of nosocomial infections but also serves as a model species of bacterial RNA biology. While its transcriptome architecture and posttranscriptional regulation through the RNA-binding proteins Hfq, RsmA, and RsmN have been studied in detail, global information about stable RNA-protein complexes in this human pathogen is currently lacking. Here, we implement gradient profiling by sequencing (Grad-seq) in exponentially growing P. aeruginosa cells to comprehensively predict RNA and protein complexes, based on glycerol gradient sedimentation profiles of >73% of all transcripts and ∼40% of all proteins. As to benchmarking, our global profiles readily reported complexes of stable RNAs of P. aeruginosa, including 6S RNA with RNA polymerase and associated product RNAs (pRNAs). We observe specific clusters of noncoding RNAs, which correlate with Hfq and RsmA/N, and provide a first hint that P. aeruginosa expresses a ProQ-like FinO domain-containing RNA-binding protein. To understand how biological stress may perturb cellular RNA/protein complexes, we performed Grad-seq after infection by the bacteriophage ΦKZ. This model phage, which has a well-defined transcription profile during host takeover, displayed efficient translational utilization of phage mRNAs and tRNAs, as evident from their increased cosedimentation with ribosomal subunits. Additionally, Grad-seq experimentally determines previously overlooked phage-encoded noncoding RNAs. Taken together, the Pseudomonas protein and RNA complex data provided here will pave the way to a better understanding of RNA-protein interactions during viral predation of the bacterial cell. IMPORTANCE Stable complexes by cellular proteins and RNA molecules lie at the heart of gene regulation and physiology in any bacterium of interest. It is therefore crucial to globally determine these complexes in order to identify and characterize new molecular players and regulation mechanisms. Pseudomonads harbor some of the largest genomes known in bacteria, encoding ∼5,500 different proteins. Here, we provide a first glimpse on which proteins and cellular transcripts form stable complexes in the human pathogen Pseudomonas aeruginosa. We additionally performed this analysis with bacteria subjected to the important and frequently encountered biological stress of a bacteriophage infection. We identified several molecules with established roles in a variety of cellular pathways, which were affected by the phage and can now be explored for their role during phage infection. Most importantly, we observed strong colocalization of phage transcripts and host ribosomes, indicating the existence of specialized translation mechanisms during phage infection. All data are publicly available in an interactive and easy to use browser. KW - Grad-seq KW - Pseudomonas KW - UKZ KW - bacteriophage KW - infection KW - Pseudomonas aeruginosa KW - RNA-binding proteins KW - noncoding RNA KW - phage Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259054 VL - 12 IS - 1 ER - TY - JOUR A1 - Doryab, Ali A1 - Taskin, Mehmet Berat A1 - Stahlhut, Philipp A1 - Schröppel, Andreas A1 - Wagner, Darcy E. A1 - Groll, Jürgen A1 - Schmid, Otmar T1 - A Biomimetic, Copolymeric Membrane for Cell‐Stretch Experiments with Pulmonary Epithelial Cells at the Air‐Liquid Interface JF - Advanced Functional Materials N2 - Chronic respiratory diseases are among the leading causes of death worldwide, but only symptomatic therapies are available for terminal illness. This in part reflects a lack of biomimetic in vitro models that can imitate the complex environment and physiology of the lung. Here, a copolymeric membrane consisting of poly(ε‐)caprolactone and gelatin with tunable properties, resembling the main characteristics of the alveolar basement membrane is introduced. The thin bioinspired membrane (≤5 μm) is stretchable (up to 25% linear strain) with appropriate surface wettability and porosity for culturing lung epithelial cells under air–liquid interface conditions. The unique biphasic concept of this membrane provides optimum characteristics for initial cell growth (phase I) and then switch to biomimetic properties for cyclic cell‐stretch experiments (phase II). It is showed that physiologic cyclic mechanical stretch improves formation of F‐actin cytoskeleton filaments and tight junctions while non‐physiologic over‐stretch induces cell apoptosis, activates inflammatory response (IL‐8), and impairs epithelial barrier integrity. It is also demonstrated that cyclic physiologic stretch can enhance the cellular uptake of nanoparticles. Since this membrane offers considerable advantages over currently used membranes, it may lead the way to more biomimetic in vitro models of the lung for translation of in vitro response studies into clinical outcome. KW - alveolar‐capillary barrier KW - cyclic mechanical stretch KW - hybrid polymers KW - in vitro cell‐stretch model KW - tunable ultra‐thin biphasic membrane Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-225645 VL - 31 IS - 10 ER - TY - JOUR A1 - Gergs, Ulrich A1 - Jahn, Tina A1 - Schulz, Nico A1 - Großmann, Claudia A1 - Rueckschloss, Uwe A1 - Demus, Uta A1 - Buchwalow, Igor B. A1 - Neumann, Joachim T1 - Protein phosphatase 2A improves cardiac functional response to ischemia and sepsis JF - International Journal of Molecular Sciences N2 - Reversible protein phosphorylation is a posttranslational modification of regulatory proteins involved in cardiac signaling pathways. Here, we focus on the role of protein phosphatase 2A (PP2A) for cardiac gene expression and stress response using a transgenic mouse model with cardiac myocyte-specific overexpression of the catalytic subunit of PP2A (PP2A-TG). Gene and protein expression were assessed under basal conditions by gene chip analysis and Western blotting. Some cardiac genes related to the cell metabolism and to protein phosphorylation such as kinases and phosphatases were altered in PP2A-TG compared to wild type mice (WT). As cardiac stressors, a lipopolysaccharide (LPS)-induced sepsis in vivo and a global cardiac ischemia in vitro (stop-flow isolated perfused heart model) were examined. Whereas the basal cardiac function was reduced in PP2A-TG as studied by echocardiography or as studied in the isolated work-performing heart, the acute LPS- or ischemia-induced cardiac dysfunction deteriorated less in PP2A-TG compared to WT. From the data, we conclude that increased PP2A activity may influence the acute stress tolerance of cardiac myocytes. KW - protein phosphorylation KW - PP2A KW - transgenic mice KW - heart KW - LPS KW - sepsis KW - ischemia Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284035 SN - 1422-0067 VL - 23 IS - 9 ER - TY - JOUR A1 - Holtfrerich, Sarah K. C. A1 - Schwarz, Katharina A. A1 - Sprenger, Christian A1 - Reimers, Luise A1 - Diekhof, Esther K. T1 - Endogenous Testosterone and Exogenous Oxytocin Modulate Attentional Processing of Infant Faces JF - PLoS ONE N2 - Evidence indicates that hormones modulate the intensity of maternal care. Oxytocin is known for its positive influence on maternal behavior and its important role for childbirth. In contrast, testosterone promotes egocentric choices and reduces empathy. Further, testosterone decreases during parenthood which could be an adaptation to increased parental investment. The present study investigated the interaction between testosterone and oxytocin in attentional control and their influence on attention to baby schema in women. Higher endogenous testosterone was expected to decrease selective attention to child portraits in a face-in-the-crowd-paradigm, while oxytocin was expected to counteract this effect. As predicted, women with higher salivary testosterone were slower in orienting attention to infant targets in the context of adult distractors. Interestingly, reaction times to infant and adult stimuli decreased after oxytocin administration, but only in women with high endogenous testosterone. These results suggest that oxytocin may counteract the adverse effects of testosterone on a central aspect of social behavior and maternal caretaking. KW - maternal behavior KW - oxytocin KW - testosterone KW - attention KW - infant faces Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-166783 VL - 11 IS - 11 ER - TY - JOUR A1 - Ryma, Matthias A1 - Tylek, Tina A1 - Liebscher, Julia A1 - Blum, Carina A1 - Fernandez, Robin A1 - Böhm, Christoph A1 - Kastenmüller, Wolfgang A1 - Gasteiger, Georg A1 - Groll, Jürgen T1 - Translation of collagen ultrastructure to biomaterial fabrication for material-independent but highly efficient topographic immunomodulation JF - Advanced materials N2 - Supplement-free induction of cellular differentiation and polarization solely through the topography of materials is an auspicious strategy but has so far significantly lagged behind the efficiency and intensity of media-supplementation-based protocols. Consistent with the idea that 3D structural motifs in the extracellular matrix possess immunomodulatory capacity as part of the natural healing process, it is found in this study that human-monocyte-derived macrophages show a strong M2a-like prohealing polarization when cultured on type I rat-tail collagen fibers but not on collagen I films. Therefore, it is hypothesized that highly aligned nanofibrils also of synthetic polymers, if packed into larger bundles in 3D topographical biomimetic similarity to native collagen I, would induce a localized macrophage polarization. For the automated fabrication of such bundles in a 3D printing manner, the strategy of “melt electrofibrillation” is pioneered by the integration of flow-directed polymer phase separation into melt electrowriting and subsequent selective dissolution of the matrix polymer postprocessing. This process yields nanofiber bundles with a remarkable structural similarity to native collagen I fibers, particularly for medical-grade poly(ε-caprolactone). These biomimetic fibrillar structures indeed induce a pronounced elongation of human-monocyte-derived macrophages and unprecedentedly trigger their M2-like polarization similar in efficacy as interleukin-4 treatment. KW - biofabrication KW - extracellular matrix KW - immunomodulation KW - macrophages KW - melt electrofibrillation KW - melt electrowriting Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-256381 VL - 33 IS - 33 ER - TY - JOUR A1 - Wasmus, Christina A1 - Dudek, Jan T1 - Metabolic Alterations Caused by Defective Cardiolipin Remodeling in Inherited Cardiomyopathies JF - Life N2 - The heart is the most energy-consuming organ in the human body. In heart failure, the homeostasis of energy supply and demand is endangered by an increase in cardiomyocyte workload, or by an insufficiency in energy-providing processes. Energy metabolism is directly associated with mitochondrial redox homeostasis. The production of toxic reactive oxygen species (ROS) may overwhelm mitochondrial and cellular ROS defense mechanisms in case of heart failure. Mitochondria are essential cell organelles and provide 95% of the required energy in the heart. Metabolic remodeling, changes in mitochondrial structure or function, and alterations in mitochondrial calcium signaling diminish mitochondrial energy provision in many forms of cardiomyopathy. The mitochondrial respiratory chain creates a proton gradient across the inner mitochondrial membrane, which couples respiration with oxidative phosphorylation and the preservation of energy in the chemical bonds of ATP. Akin to other mitochondrial enzymes, the respiratory chain is integrated into the inner mitochondrial membrane. The tight association with the mitochondrial phospholipid cardiolipin (CL) ensures its structural integrity and coordinates enzymatic activity. This review focuses on how changes in mitochondrial CL may be associated with heart failure. Dysfunctional CL has been found in diabetic cardiomyopathy, ischemia reperfusion injury and the aging heart. Barth syndrome (BTHS) is caused by an inherited defect in the biosynthesis of cardiolipin. Moreover, a dysfunctional CL pool causes other types of rare inherited cardiomyopathies, such as Sengers syndrome and Dilated Cardiomyopathy with Ataxia (DCMA). Here we review the impact of cardiolipin deficiency on mitochondrial functions in cellular and animal models. We describe the molecular mechanisms concerning mitochondrial dysfunction as an incitement of cardiomyopathy and discuss potential therapeutic strategies. KW - cardiolipin KW - mitochondria KW - Barth syndrome KW - Sengers syndrome KW - respiratory chain KW - Dilated Cardiomyopathy with Ataxia KW - cardiomyopathy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219286 SN - 2075-1729 VL - 10 IS - 11 ER - TY - JOUR A1 - Kade, Juliane C. A1 - Bakirci, Ezgi A1 - Tandon, Biranche A1 - Gorgol, Danila A1 - Mrlik, Miroslav A1 - Luxenhofer, Robert A1 - Dalton, Paul D. T1 - The Impact of Including Carbonyl Iron Particles on the Melt Electrowriting Process JF - Macromolecular Materials and Engineering N2 - Melt electrowriting, a high-resolution additive manufacturing technique, is used in this study to process a magnetic polymer-based blend for the first time. Carbonyl iron (CI) particles homogenously distribute into poly(vinylidene fluoride) (PVDF) melts to result in well-defined, highly porous structures or scaffolds comprised of fibers ranging from 30 to 50 µm in diameter. This study observes that CI particle incorporation is possible up to 30 wt% without nozzle clogging, albeit that the highest concentration results in heterogeneous fiber morphologies. In contrast, the direct writing of homogeneous PVDF fibers with up to 15 wt% CI is possible. The fibers can be readily displaced using magnets at concentrations of 1 wt% and above. Combined with good viability of L929 CC1 cells using Live/Dead imaging on scaffolds for all CI concentrations indicates that these formulations have potential for the usage in stimuli-responsive applications such as 4D printing. KW - additive manufacturing KW - melt electrospinning writing KW - magnetoactive materials KW - electroactive polymers Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-318482 SN - 1438-7492 VL - 307 IS - 12 ER - TY - JOUR A1 - Schmidt, Enno A1 - Sticherling, Michael A1 - Sárdy, Miklós A1 - Eming, Rüdiger A1 - Goebeler, Matthias A1 - Hertl, Michael A1 - Hofmann, Silke C. A1 - Hunzelmann, Nicolas A1 - Kern, Johannes S. A1 - Kramer, Harald A1 - Nast, Alexander A1 - Orzechowski, Hans‐Dieter A1 - Pfeiffer, Christiane A1 - Schuster, Volker A1 - Sitaru, Cassian A1 - Zidane, Miriam A1 - Zillikens, Detlef A1 - Worm, Margitta T1 - S2k guidelines for the treatment of pemphigus vulgaris/foliaceus and bullous pemphigoid: 2019 update JF - JDDG: Journal der Deutschen Dermatologischen Gesellschaft KW - pemphigus vulgaris KW - pemphigus foliaceus KW - S2k guidelines Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-217806 VL - 18 IS - 5 SP - 516 EP - 526 ER - TY - JOUR A1 - Moratin, Helena A1 - Thöle, Anna A1 - Lang, Josephine A1 - Ehret Kasemo, Totta A1 - Stöth, Manuel A1 - Hagen, Rudolf A1 - Scherzad, Agmal A1 - Hackenberg, Stephan T1 - Ag- but not ZnO-nanoparticles disturb the airway epithelial barrier at subtoxic concentrations JF - Pharmaceutics N2 - Inhalation is considered to be the most relevant source of human exposure to nanoparticles (NPs); however, only a few investigations have addressed the influence of exposing the respiratory mucosal barrier to subcytotoxic doses. In the nasal respiratory epithelium, cells of the mucosa represent one of the first contact points of the human organism with airborne NPs. Disruption of the epithelial barrier by harmful materials can lead to inflammation in addition to potential intrinsic toxicity of the particles. The aim of this study was to investigate whether subtoxic concentrations of zinc oxide (ZnO)- and silver (Ag)-NPs have an influence on upper airway barrier integrity. Nasal epithelial cells from 17 donors were cultured at the air–liquid interface and exposed to ZnO- and Ag-NPs. Barrier function, quantified by transepithelial electrical resistance (TEER), decreased after treatment with 10 µg/mL Ag-NPs, but FITC-dextran permeability remained stable and no change in mRNA levels of tight junction proteins and E-cadherin was detected by real-time quantitative PCR (RT-qPCR). The results indicate that subtoxic concentrations of Ag-NPs may already induce damage of the upper airway epithelial barrier in vitro. The lack of similar disruption by ZnO-NPs of similar size suggests a specific effect by Ag-NPs. KW - epithelial barrier KW - nanoparticles KW - tight junctions KW - zinc oxide KW - silver Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-357403 SN - 1999-4923 VL - 15 IS - 10 ER - TY - JOUR A1 - Israel, Ina A1 - Riehl, Gabriele A1 - Butt, Elke A1 - Buck, Andreas K. A1 - Samnick, Samuel T1 - Gallium-68-labeled KISS1-54 peptide for mapping KISS1 receptor via PET: initial evaluation in human tumor cell lines and in tumor-bearing mice JF - Pharmaceuticals N2 - Kisspeptins (KPs, KISS1) and their receptor (KISS1R) play a pivotal role as metastasis suppressor for many cancers. Low or lost KP expression is associated with higher tumor grade, increased metastatic potential, and poor prognosis. Therefore, KP expression has prognostic relevance and correlates with invasiveness in cancers. Furthermore, KISS1R represents a very promising target for molecular imaging and therapy for KISS1R-expressing tumors. The goal of this study was to evaluate the developed KISS1-54 derivative, [\(^{68}\)Ga]KISS1-54, as a PET-imaging probe for KISS1R-expressing tumors. The NODAGA-KISS1-54 peptide was labeled by Gallium-68, and the stability of the resulting [\(^{68}\)Ga]KISS1-54 evaluated in injection solution and human serum, followed by an examination in different KISS1R-expressing tumor cell lines, including HepG2, HeLa, MDA-MB-231, MCF7, LNCap, SK-BR-3, and HCT116. Finally, [\(^{68}\)Ga]KISS1-54 was tested in LNCap- and MDA-MB-231-bearing mice, using µ-PET, assessing its potential as an imaging probe for PET. [\(^{68}\)Ga]KISS1-54 was obtained in a 77 ± 7% radiochemical yield and at a >99% purity. The [\(^{68}\)Ga]KISS1-54 cell uptake amounted to 0.6–4.4% per 100,000 cells. Moreover, the accumulation of [\(^{68}\)Ga]KISS1-54 was effectively inhibited by nonradioactive KISS1-54. In [\(^{68}\)Ga]KISS1-54-PET, KISS1R-positive LNCap-tumors were clearly visualized as compared to MDA-MB-231-tumor implant with predominantly intracellular KISS1R expression. Our first results suggest that [\(^{68}\)Ga]KISS1-54 is a promising candidate for a radiotracer for targeting KISS1R-expressing tumors via PET. KW - [\(^{68}\)]KISS1-54 KW - KISS1 receptor KW - GPR54 KW - kisspeptin KW - human tumor cell lines KW - positron emission tomography KW - PET KW - KISS1-54 Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-355898 SN - 1424-8247 VL - 17 IS - 1 ER - TY - JOUR A1 - Odorfer, Thorsten M. A1 - Volkmann, Jens T1 - Deep brain stimulation for focal or segmental craniocervical dystonia in patients who have failed botulinum neurotoxin therapy - a narrative review of the literature JF - Toxins N2 - (1) Background: The first-line treatment for patients with focal or segmental dystonia with a craniocervical distribution is still the intramuscular injection of botulinum neurotoxin (BoNT). However, some patients experience primary or secondary treatment failure from this potential immunogenic therapy. Deep brain stimulation (DBS) may then be used as a backup strategy in this situation. (2) Methods: Here, we reviewed the current study literature to answer a specific question regarding the efficacy and safety of the use of DBS, particularly for cervical dystonia (CD) and Meige syndrome (MS) in patients with documented treatment failure under BoNT. (3) Results: There are only two studies with the highest level of evidence in this area. Despite this clear limitation, in the context of the narrowly defined research question of this paper, it is possible to report 161 patients with CD or MS who were included in studies that were able to show a statistically significant reduction in dystonic symptoms using DBS. Safety and tolerability data appeared adequate. However, much of the information is based on retrospective observations. (4) Conclusions: The evidence base in this area is in need of further scientific investigation. Most importantly, more randomized, controlled and double-blind trials are needed, possibly including a head-to-head comparison of DBS and BoNT. KW - cervical dystonia KW - Meige syndrome KW - deep brain stimulation KW - internal globus pallidus KW - subthalamic nucleus KW - botulinum neurotoxin KW - medication therapy failure KW - symptom control KW - safety and tolerability Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-357707 SN - 2072-6651 VL - 15 IS - 10 ER - TY - JOUR A1 - Watzling, Martin A1 - Klaus, Lorenz A1 - Weidemeier, Tamara A1 - Horder, Hannes A1 - Ebert, Regina A1 - Blunk, Torsten A1 - Bauer-Kreisel, Petra T1 - Three-dimensional breast cancer model to investigate CCL5/CCR1 expression mediated by direct contact between breast cancer cells and adipose-derived stromal cells or adipocytes JF - Cancers N2 - The tumor microenvironment (TME) in breast cancer is determined by the complex crosstalk of cancer cells with adipose tissue-inherent cells such as adipose-derived stromal cells (ASCs) and adipocytes resulting from the local invasion of tumor cells in the mammary fat pad. This leads to heterotypic cellular contacts between these cell types. To adequately mimic the specific cell-to-cell interaction in an in vivo-like 3D environment, we developed a direct co-culture spheroid model using ASCs or differentiated adipocytes in combination with MDA-MB-231 or MCF-7 breast carcinoma cells. Co-spheroids were generated in a well-defined and reproducible manner in a high-throughput process. We compared the expression of the tumor-promoting chemokine CCL5 and its cognate receptors in these co-spheroids to indirect and direct standard 2D co-cultures. A marked up-regulation of CCL5 and in particular the receptor CCR1 with strict dependence on cell–cell contacts and culture dimensionality was evident. Furthermore, the impact of direct contacts between ASCs and tumor cells and the involvement of CCR1 in promoting tumor cell migration were demonstrated. Overall, these results show the importance of direct 3D co-culture models to better represent the complex tumor–stroma interaction in a tissue-like context. The unveiling of tumor-specific markers that are up-regulated upon direct cell–cell contact with neighboring stromal cells, as demonstrated in the 3D co-culture spheroids, may represent a promising strategy to find new targets for the diagnosis and treatment of invasive breast cancer. KW - 3D breast cancer model KW - adipose-derived stromal cells KW - adipocytes KW - adipose tissue KW - spheroids KW - co-culture Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-362502 SN - 2072-6694 VL - 15 IS - 13 ER - TY - THES A1 - Stark, Irmgard Katharina T1 - Einfluss von Interferon auf das Infektionsverhalten von Herpes simplex Virus 1 und seiner DUB - Mutante C65A in der Zellkultur T1 - The influence of interferon on infection of Herpes simplex Virus 1 and its DUB – mutant C65A in cell culture N2 - Die Erforschung viraler Proteine ist wichtig, um virale Infektionen besser verstehen und damit therapieren zu können. Die Aufklärung der DUB-Funktion auf dem viralen Herpesprotein pUL36 ermöglicht ein besseres Verständnis des Infektionshergangs und könnte zur Entwicklung eines Enzyminhibitors führen, der nur an diesem Enzym ansetzt, nachdem es sich von den zellulären DUBs unterscheidet (Kattenhorn et al., 2005). In dieser Arbeit konnten die vorherigen Daten, die eine stärkere Hemmung der DUB- Mutante unter Interferoneinfluss zeigten, in unterschiedlichen Assay-Designs bestätigt werden. Auch Versuche mit einem anderen Herpes simplex Virus Strang, bestätigten die vorherigen Daten. Die Ergebnisse zeigen, dass die DUB-Funktion für HSV-1 wichtig ist für die virale Evasion der zellulären Immunantwort. Die genaue Funktion der DUB in der Infektion ist jedoch unklar. Aufgrund der vorbestehenden Datenlage erschien am wahrscheinlichsten, dass die DUB-Funktion vor Eindringen des Herpes Simplex Virus in den Zellkern zum Tragen kommt, womit es nach Abnahme des Interferons nicht zu einer viralen Reaktivierung käme. Deshalb wurden Untersuchungen unternommen, um eine mögliche Reaktivierung nach Abnahme des Interferons näher zu untersuchen. Hierfür wurden zwei verschiedene Experimente entwickelt. Einmal wurde das Interferon direkt nach Infektion und einmal 3 Tage nach Infektion (3dpi) abgenommen. Die Ergebnisse zeigten beide eine stärkere Hemmung der DUB-HSV-1-Mutante unter Interferoneinfluss. Bei Abnahme des Interferons direkt nach Infektion lag bei Wildtyp und Mutante ein leichter Anstieg der Plaquezahlen vor, wobei dieser Effekt von der Dosis des Interferons abhängig war. Eine hohe Interferondosis begünstigte bei beiden eine stärkere Hemmung, allerdings bei beiden auch eine leichte Erhöhung der Plaquezahl nach Abnahme. Bei einer niedrigen Dosis konnte nur eine stärkere Hemmung der DUB-Mutante, jedoch keine Reaktivierung bei Wildtyp und Mutante nach Abnahme des Interferons gezeigt werden. Bei Abnahme drei Tage nach Infektion zeigte sich sowohl bei dem Wildtyp-Virus als auch der DUB- Mutante kein Anstieg in den Plaquezahlen. Es sind, nachdem Deubiquitinierung nicht nur eine Rolle in der Verhinderung des proteosomalen Abbaus von in die Zelle eingedrungenem Virus spielt, sondern auch der Zellregulation, mehrere Szenarien denkbar, die diesen Phänotyp erklären könnten. Die DUB-Funktion könnte zwar den proteosomalen Abbau durch Deubiqutinierung und damit Verhinderung der Markierung des Virus zum zellulären Abbau verhindern. Allerdings könnten sich durch einen langsameren Transport aus der Zelle oder in den Nucleus auch weniger Plaques bei der Mutante als wie beim Wildtyp unter Interferoneinfluss bilden, nachdem das Virus dann leichter Ziel antiviraler Proteine werden könnte. Oder die DUB-Funktion spielt eine Rolle beim Eintritt in den Kern durch Modifikationen anderer Proteine. Virengenome könnten auch durch eine fehlende DUB-Funktion reprimiert werden oder die Zelle durch Apoptose absterben. Interessanterweise konnte keine Hemmung der DUB-Mutante in Interferon behandelten U-2 OS Zellen gezeigt werden, von denen ein Defekt im STING- vermittelten Signalweg bekannt ist. Vielleicht zeigt dies, dass das STING-Protein an dem gezeigten DUB-Phänotyp beteiligt ist. Nachgewiesen ist außerdem bereits eine Funktion des Enzyms bei der zweiten Umhüllung der Kapside bei Pseudorabiesvirus (Möhl, 2011). Weitere Untersuchungen unter Einsatz bspw. von Immunfluoreszenz, Proteasominhibitoren oder weiteren Zelllinien wie Saos-2, sind nötig, um die genaue Funktion zu klären. N2 - The study of viral proteins is important to better understand and thus treat viral infections. Elucidation of DUB function on the viral herpes protein pUL36 provides a better understanding of the infection process and could lead to the development of an enzyme inhibitor that targets only this enzyme after it is different from cellular DUBs (Kattenhorn et al., 2005). In this work, previous data showing greater inhibition of the DUB- mutant under interferon influence were confirmed in different assay designs. Also, experiments with a different herpes simplex virus strand, confirmed the previous data. The results indicate that DUB function for HSV-1 is important for viral evasion of the cellular immune response. However, the exact function of DUB in infection is unclear. Based on the preexisting data, it seemed most likely that DUB function would come into play before herpes simplex virus enters the nucleus, which would mean that viral reactivation would not occur after interferon depletion. Therefore, studies were undertaken to further investigate a possible reactivation after decrease of interferon. Two different experiments were developed for this purpose. Once the interferon was withdrawn immediately after infection and once 3 days after infection (3dpi). The results both showed a stronger inhibition of the DUB-HSV-1 mutant under interferon influence. When interferon was decreased immediately after infection, a slight increase in plaque counts was present in both wild type and mutant, although this effect was dependent on the dose of interferon. A high dose of interferon promoted greater inhibition in both, but also a slight increase in plaque numbers after decrease in both. A low dose showed only greater inhibition of the DUB mutant but no reactivation in wild type and mutant after decrease of interferon. When decreased three days after infection, there was no increase in plaque counts for either the wild-type virus or the DUB- mutant. Given that deubiquitination plays a role not only in preventing proteosomal degradation of virus that has entered the cell but also in cell regulation, several scenarios are conceivable that could explain this phenotype. To be sure, DUB function could prevent proteosomal degradation by deubiqutinating and thereby preventing the virus from being labeled for cellular degradation. However, slower transport out of the cell or into the nucleus could also result in fewer plaques forming in the mutant than in the wild type under interferon influence, after which the virus could more easily become a target of antiviral proteins. Alternatively, DUB function may play a role in entry into the nucleus through modifications of other proteins. Viral genomes could also be repressed by a lack of DUB function or the cell could die by apoptosis. Interestingly, no inhibition of the DUB mutant was shown in interferon-treated U-2 OS cells, which are known to have a defect in the STING-mediated signaling pathway. Perhaps this indicates that the STING protein is involved in the DUB phenotype shown. Furthermore, a function of the enzyme in the second envelope of capsids in pseudorabies virus has already been demonstrated (Möhl, 2011). Further studies using e.g. immunofluorescence, proteasome inhibitors or additional cell lines such as Saos-2, are necessary to clarify the exact function. KW - Herpes simplex Virus DUB C65A KW - DUB Mutante KW - Herpes simplex virus C65A KW - Interferon KW - Zellkultur Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-351950 ER - TY - JOUR A1 - Güder, Gülmisal A1 - Rein, Eva von A1 - Flohr, Thomas A1 - Weismann, Dirk A1 - Schmitt, Dominik A1 - Störk, Stefan A1 - Frantz, Stefan A1 - Kratzer, Vincent A1 - Kendi, Christian T1 - Motion detectors as additional monitoring devices in the intensive care unit — a proof-of-concept study JF - Applied Sciences N2 - Background: Monitoring the vital signs of delirious patients in an intensive care unit (ICU) is challenging, as they might (un-)intentionally remove devices attached to their bodies. In mock-up scenarios, we systematically assessed whether a motion detector (MD) attached to the bed may help in identifying emergencies. Methods: We recruited 15 employees of the ICU and equipped an ICU bed with an MD (IRON Software GmbH, Grünwald, Germany). Participants were asked to replay 22 mock-up scenes of one-minute duration each: 12 scenes with movements and 10 without movements, of which 5 were emergency scenes (“lying dead-still, with no or very shallow breathing”). Blinded recordings were presented to an evaluation panel consisting of an experienced ICU nurse and a physician, who was asked to assess and rate the presence of motions. Results: Fifteen participants (nine women; 173 ± 7.0 cm; 78 ± 19 kg) joined the study. In total, 286 out of 330 scenes (86.7%) were rated correctly. Ratings were false negative (FN: “no movements detected, but recorded”) in 7 out of 180 motion scenes (3.9%). Ratings were false positive (FP: “movements detected, but not recorded”) in 37 out of 150 scenes (24.7%), more often in men than women (26 out of 60 vs. 11 out of 90, respectively; p < 0.001). Of note, in 16 of these 37 FP-rated scenes, a vibrating mobile phone was identified as a potential confounder. The emergency scenes were correctly rated in 64 of the 75 runs (85.3%); 10 of the 11 FP-rated scenes occurred in male subjects. Conclusions: The MD allowed for identifying motions of test subjects with high sensitivity (96%) and acceptable specificity (75%). Accuracy might increase further if activities are recorded continuously under real-world conditions. KW - motion detector KW - noncontact monitoring KW - Internet of Things devices Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-362404 SN - 2076-3417 VL - 13 IS - 16 ER - TY - JOUR A1 - Hiew, Shawn A1 - Eibeck, Leila A1 - Nguemeni, Carine A1 - Zeller, Daniel T1 - The influence of age and physical activity on locomotor adaptation JF - Brain Sciences N2 - Background: Aging increases individual susceptibility to falls and injuries, suggesting poorer adaptation of balance responses to perturbation during locomotion, which can be measured with the locomotor adaptation task (LAT). However, it is unclear how aging and lifestyle factors affect these responses during walking. Hence, the present study investigates the relationship between balance and lifestyle factors during the LAT in healthy individuals across the adult lifespan using a correlational design. Methods: Thirty participants aged 20–78 years performed an LAT on a split-belt treadmill (SBT). We evaluated the magnitude and rate of adaptation and deadaptation during the LAT. Participants reported their lifelong physical and cognitive activity. Results: Age positively correlated with gait-line length asymmetry at the late post-adaptation phase (p = 0.007). These age-related effects were mediated by recent physical activity levels (p = 0.040). Conclusion: Our results confirm that locomotor adaptive responses are preserved in aging, but the ability to deadapt newly learnt balance responses is compromised with age. Physical activity mediates these age-related effects. Therefore, gait symmetry post-adaptation could effectively measure the risk of falling, and maintaining physical activity could protect against declines in balance. KW - locomotor adaptation KW - walking KW - physical activity KW - exercise KW - aging KW - balance Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-362478 SN - 2076-3425 VL - 13 IS - 9 ER - TY - THES A1 - Keicher, Franca T1 - Muskuläre Einflüsse der Rotatorenmanschette auf die Early Onset Arthrose der Schulter T1 - Muscular Influences of the Rotator Cuff on Early Onset Shoulder Osteoarthritis N2 - Introduction: The etiology of early onset shoulder arthritis (EOA) remains unclear. Due to the influence of the muscles of the rotator cuff (RC) on glenohumeral kinematics, muscular causes are being discussed. The aim of the study was to identify corresponding factors on EOA based on volume measurements of the RC and examinations of the adjacent bony structures in MRI imaging, as well as the collection of patient-specific characteristics. Methods: In a case-control study, shoulders of 15 patients (14 men, 1 woman) with shoulder arthritis before the age of 60 and 13 control subjects (13 men) were examined. Anthropometric body measurements and clinical characteristics were collected. The volumes of the RC were calculated using manual tracing of the individual muscle cross- sections on MRI scans. In addition, the angles between the coracoid or scapular spine and scapular body were measured. The position of the scapula to the thorax was determined by recording the angle of the scapula to the plane of the table. The glenoid retroversion, morphological types, and humeral head subluxation were also collected. The analysis was conducted using descriptive and comparative statistical methods, as well as logistic and linear regression analyses. Results: The volumes of the RC did not differ significantly between the patients with EOA and the control subjects, either in absolute terms or in relative proportions. However, significantly higher values of thorax circumference and diameter, body weight, and BMI were found in the diseased individuals compared to the controls. Furthermore, these individuals were significantly more likely to have occupations that expended more than 1400 kcal per day. The risk of EOA increased with the steeper angle of the scapula to the thorax. Patients with B2-glenoid had significantly larger angles between the coracoid and scapular body as well as higher values of transverse thorax diameter, than those with B1-glenoid. Regardless of whether EOA was present or not, engaging in overhead sports was associated with a higher subluxation index. Conclusion: While the RC did not show any abnormalities in EOA patients, male gender, BMI, thoracic shape and scapular position, as well as certain occupations and sports, were associated with EOA. Further studies are needed to investigate these risk factors in EOA more precisely and develop possible treatment concepts. Peripheral shoulder muscles (such as teres major or pectoralis major) should also be included in the investigations. N2 - Einleitung: Die Ätiologie der Early Onset Arthrose (EOA) der Schulter ist bislang ungeklärt. Aufgrund des Einflusses der Muskeln der Rotatorenmanschette (RM) auf die glenohumerale Kinematik werden muskuläre Ursachen diskutiert. Ziel der Studie war es, entsprechende Faktoren anhand von Volumenmessungen der RM und Untersuchungen der angrenzenden knöchernen Strukturen in der MRT-Bildgebung sowie der Erfassung patientenspezifischer Charakteristika zu identifizieren. Methoden: In einer Fall-Kontroll-Studie wurden Schultern von 15 PatientInnen (14 Männer, 1 Frau) mit einer Omarthrose vor dem 60. Lebensjahr und 13 Kontrollpersonen (ausschließlich Männer) untersucht. Dabei wurden anthropometrische Körpermessungen sowie klinische Charakteristika erhoben. Anhand von MRT-Bildern wurden die Volumina der RM mittels manueller Umrandung der einzelnen Muskelquerschnitte berechnet. Weiterhin wurden die Winkel zwischen Coracoid bzw. Spina scapulae und Scapulakörper gemessen. Die Stellung der Scapula zum Thorax wurde anhand des Winkels der Scapula zur Tischebene erhoben. Zudem wurden die Glenoidretroversion, -morphologietypen und Humeruskopfsubluxation ermittelt. Die Auswertung erfolgte anhand deskriptiver und vergleichender statistischer Verfahren sowie logistischer und linearer Regressionsanalysen. Ergebnisse: Die Volumina der RM von PatientInnen mit EOA unterschieden sich weder absolut noch im relativen Verhältnis von denen der Kontrollpersonen. Dagegen konnten im Vergleich signifikant höhere Werte des Thoraxumfangs und -durchmessers, des Körpergewichts sowie des BMI bei erkrankten Personen nachgewiesen werden. Weiterhin übten diese signifikant häufiger Berufe aus, die mehr als 1400 kcal pro Tag verbrauchten. Das Risiko einer EOA nahm zu, je steiler die Scapula zum Thorax stand. PatientInnen mit B2-Glenoid wiesen signifikant größere Winkel zwischen Coracoid und Scapulakörper sowie höhere Werte des transversalen Thoraxdurchmessers auf, als erkrankte Personen mit einem B1-Glenoid. Unabhängig von einer Erkrankung mit EOA zeigte sich, dass das Ausüben einer Überkopfsportart mit einem höheren Subluxationsindex assoziiert war. Fazit: Während die RM keine Auffälligkeiten bei PatientInnen mit EOA zeigte, waren ein männliches Geschlecht, der BMI, die Thoraxform und Scapulastellung, sowie die Ausübung bestimmter Berufe und Sportarten mit der Erkrankung assoziiert. Weitere Studien müssen folgen, um diese Risikofaktoren bei EOA genauer zu untersuchen und mögliche Therapiekonzepte zu entwickeln. Dabei sollte auch die periphere Schultermuskulatur (z. B. M. teres major oder M. pectoralis major) in die Untersuchungen miteinbezogen werden. KW - Arthrose KW - Schultergelenk KW - Early Onset Schulterarthrose KW - Rotatorenmanschette Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360557 ER - TY - THES A1 - Rehlinghaus, Christine T1 - Retrospektive Evaluation der intravenösen Dexamethason- bzw. Methylprednisolon-Pulstherapie bei ausgeprägter Alopecia areata T1 - Retrospective evaluation of intravenous dexamethasone or methylprednisolone pulse therapy for severe alopecia areata N2 - Hintergrund: Bei der Entscheidung für eine intravenöse Kortikosteroid-Pulstherapie bei schweren Formen der AA ist die Abwägung von Therapieaufwand, Nebenwirkungen und Risiken einerseits und der Erfolgsaussicht andererseits von zentraler Bedeutung. Ziel: Ziel dieser retrospektiven Analyse war es daher, die Wirksamkeit und Sicherheit der intravenösen Kortikosteroid-Pulstherapie bei Patient:innen mit ausgeprägter AA klinikintern als qualitätssichernde Maßnahme zu untersuchen, prognostisch bedeutsame Faktoren für den Therapieeffekt zu ermitteln und hierdurch die beste Indikation herauszuarbeiten. Methode: 126 Patient:innen (13 Kinder und Jugendliche) erhielten Dexamethason 100 mg (122 Patienten) oder Methylprednisolon 20-30 mg/kg/KG (max. 1000 mg, 4 Patienten) an drei aufeinanderfolgenden Tagen für ein bis drei Zyklen. Ergebnisse: Patienten mit einer AA partialis bzw. diffusa zeigten im Hinblick auf ein vollständiges oder kosmetisch akzeptables Wiederwachstum die besten Ansprechraten (44,3%, n=43). Unter den Ophiasis-Patienten und den Patienten mit AA totalis/universalis sprach nur etwa ein Viertel auf die Therapie an (Ophiasis 23,8%, n=5; AA totalis/universalis: 25%, n=2). Schwerwiegende unerwünschte Nebenwirkungen wurden nicht beobachtet. Schlussfolgerung: In der vorliegenden Untersuchung ließen sich eine längere Bestandsdauer der Erkrankung und Erkrankungsepisode (über 6 Monate), ein schwerer Ausprägungsgrad (Ophiasis, AA totalis/universalis) und krankheitstypische Nagelveränderungen als wichtige ungünstige prognostische Faktoren nachweisen. Dagegen wirkten sich die untersuchten Kriterien Alter, Geschlecht, atopisches Ekzem und andere Erkrankungen des atopischen Formenkreises, Schilddrüsen- und Autoimmunerkrankungen in der Eigenanamnese sowie AA in der Familienanamnese nicht negativ auf den Behandlungserfolg aus. Patienten mit AA partialis und einer Bestandsdauer der AA von maximal 6 Monaten haben die besten Erfolgsaussichten. N2 - Background: When deciding in favour of intravenous corticosteroid pulse therapy for severe forms of AA, it is of central importance to weigh up the therapeutic effort, side effects and risks on the one hand and the prospects of success on the other. Aim: The aim of this retrospective analysis was therefore to investigate the efficacy and safety of intravenous corticosteroid pulse therapy in patients with pronounced AA within the clinic as a quality assurance measure, to determine prognostically significant factors for the therapeutic effect and thus to identify the best indication. Methods: 126 patients (13 children and adolescents) received dexamethasone 100 mg (122 patients) or methylprednisolone 20-30 mg/kg/KG (max. 1000 mg, 4 patients) on three consecutive days for one to three cycles. Results: Patients with AA partialis or diffusa showed the best response rates in terms of complete or cosmetically acceptable regrowth (44.3%, n=43). Among the ophiasis patients and the patients with AA totalis/universalis, only about a quarter responded to the therapy (ophiasis 23.8%, n=5; AA totalis/universalis: 25%, n=2). No serious adverse events were observed. Conclusion: In the present study, a longer duration of the disease and disease episode (more than 6 months), a severe degree of severity (ophiasis, AA totalis/universalis) and nail changes typical of the disease were found to be important unfavourable prognostic factors. In contrast, the investigated criteria of age, gender, atopic eczema and other atopic diseases, thyroid and autoimmune diseases in the patient's own medical history and AA in the family history did not have a negative effect on the success of treatment. Patients with AA partialis and a maximum duration of AA of 6 months have the best chances of success. KW - Alopecia areata Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360711 ER - TY - THES A1 - Fieber, Tabea T1 - Retrospektive unizentrische Analyse des Komplikationsmanagements bei Anastomoseninsuffizienz nach linksseitiger Kolon- und Rektumresektion T1 - Retrospective unicenter study of management of anastomotic leakage after left hemicolectomy and rectal resection N2 - Eine gefürchtete Komplikation nach Resektionen am Kolon mit Wiederherstellung der Kontinuität ist das Auftreten einer Anastomoseninsuffizienz (AI). Der Prozess der Diagnosestellung und das therapeutische Vorgehen sind zentrumsspezifisch und sehr heterogen. Ziel dieser Promotionsarbeit war die deskriptive Darstellung der Prävalenz, Diagnostik und Therapie von AI, um anhand dieser Daten ein bestimmtes zu favorisierendes Vorgehen zur AI-Behandlung herauszuarbeiten. Es wurde eine retrospektive unizentrische Analyse durchgeführt. Diese umfasste eine Kohorte von 744 Patienten, welche von 2009 bis 2013 am Universitätsklinikum Würzburg unter Kontinuitätserhalt kolorektal reseziert wurden. Es erfolgte eine deskriptive und statistische Auswertung mittels uni- und multivariater Analysen in Bezug auf Demographie, Risikofaktoren für die Entwicklung einer AI und den Erfolg der einzelnen Therapiekonzepte. Während der Nachbeobachtungsphase vom im Mittel 2,5 Jahren entwickelten 10,48% der Patienten eine AI. 60% der Insuffizienzen wurden während der ersten 7 postoperativen Tage detektiert. Als Risikofaktoren konnten indikationsunabhängig männliches Geschlecht, offener Zugangsweg und pulmonale Erkrankungen herausgearbeitet werden. Indikationsspezifisch zeigte sich eine Zunahme des AI-Risikos bei Divertikulitis-Patienten mit pulmonalen Erkrankungen (OR 4,5) und Cortisoneinnahme (OR 5,4). Auffällig wurden Patienten mit AI durch heterogene und teils unspezifische Symptome – am häufigsten durch Fieber (28,21%) und auffällige Laborwerte (48,72%). Eine folgende CT-Diagnostik bestätigte die Diagnose in 76,32% der Fälle und war in 24,48% falsch negativ. Patienten mit schlechtem AZ bei Diagnose der AI zeigten eine signifikant höhere Mortalität. Ein protektives Stoma konnte eine AI nicht verhindern, aber ihre Symptome und die Schwere des Verlaufs abmildern. Gemessen an der Überlebensrate und der Revisionspflichtigkeit unterschieden sich die durchgeführten Maßnahmen beim Versuch der kontinuitätserhaltenden Therapie nicht in Bezug auf den Erfolg der Therapie. Wie Insuffizienz- und Mortalitätsrate nach AI zeigen, ist diese unizentrische Analyse international vergleichbar. Die Ableitung einer generellen Empfehlung zur therapeutischen Vorgehensweise bei AI ist nicht möglich. Vielmehr sind alle dargestellten Maßnahmen zur Beherrschung der AI sinnvoll, während die Wahl der Vorgehensweise weiterhin eine Individualentscheidung bleibt. N2 - Anastomotic leakage (AL) is a feared complication following continuity-preserving colon resection. The process of making the diagnosis and the planning of a therapeutic strategy are center-specific and very heterogeneous. The aim of this doctoral thesis is to describe the prevalence, diagnosis and therapy of AL, in order to use this data to identify a specific strategy that would overall improve outcomes while treating AL. A retrospective unicentric analysis was performed. This included a cohort of 744 patients who underwent colorectal resection with continuity preservation at the Würzburg University Hospital from 2009 to 2013. A descriptive and statistical evaluation was carried out using univariate and multivariate analyzes regarding demographics, risk factors for the development of AL and the success of the individual therapy concepts. During the follow-up period of a mean of 2.5 years, 10.48% of patients developed AL. 60% of the insufficiencies were detected during the first 7 postoperative days. Male gender, open access and pulmonary diseases were identified as non-surgery-related risk factors, while diverticulitis patients with pulmonary diseases (OR 4.5) and patients taking cortisone (OR 5.4) were identified as surgery-related risk factor. Patients with AL were characterized by heterogeneous and sometimes non-specific symptoms - most commonly fever (28.21%) and abnormal laboratory values (48.72%). A follow-up CT scan confirmed the diagnosis in 76.32% of cases and was false negative in 24.48%. Patients with poor general health at the time of diagnosing AL showed a significantly higher mortality rate. A protective stoma did not prevent AL, however showed to alleviate its symptoms and course severity. Measured in terms of the survival rate and the need for revision surgery, the measures carried out did not differ in success of the therapy when attempting continuity-preserving therapy. The rates of insufficiency and mortality after AL demonstrate, that this unicentric analysis is internationally comparable. It is was, however, at the time of this study not possible to derive a general recommendation for the therapeutic strategy towards AL. Rather, all of the measures presented for mastering AL make sense, while the choice of the strategy remains an individual decision. KW - Darmanastomose KW - Anastomoseninsuffizienz KW - Komplikationsmanagement KW - Rektumresektion Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360730 ER - TY - THES A1 - Enzensberger, Eva T1 - Stellenwert der Dobutamin-Stress-Echokardiographie bei der Unterscheidung einer hochgradigen von einer pseudo-hochgradigen Aortenklappenstenose und Bestimmung deren echokardiographischer Prädiktoren T1 - Value of dobutamine stress echocardiography in differentiating between true-severe and pseudo-severe low-gradient aortic stenosis and determining their echocardiographic predictors N2 - Ziel dieser Studie war es, zu eruieren, ob die DSE zur Unterscheidung einer TS- von einer PSAS beitragen kann. Ebenfalls untersuchten wir, ob es bestimmte echokardiographische Prädiktoren für eine TS- und eine PSAS gibt und ob die LVEF bei Patienten mit einer LGAS eine entscheidende Rolle spielt. Methoden: Es wurde bei 130 Patienten mit einer asymptomatischen AS im Uniklinikum Würzburg zwischen Januar 2011 und Dezember 2016 sowohl eine TTE als auch eine DSE durchgeführt. Mittels TTE wurden verschiedene echokardiographische Daten erhoben und falls die Patienten eine AVAi  0,6 cm2/m2 und eine PGmean < 40 mmHg aufwiesen, wurden sie in die Studie eingeschlossen. Sie wurden in zwei Gruppen aufgeteilt, je nachdem ob sie eine LGAS mit einer LVEF  50% oder < 50% aufwiesen. Bei allen Patienten wurde in der DSE die AVAproj berechnet und sie wurden daraufhin in zwei Untergruppen aufgeteilt, Patienten mit einer AVAproj  1 cm2 wurden der Gruppe mit einer hochgradigen LGAS (TS-LGAS) und Patienten mit einer AVAproj > 1cm2 der Gruppe mit einer pseudo-hochgradigen LGAS (PS-LGAS) zugeteilt. Alle Patientendaten wurden manuell ausgewertet. Das klinische Follow Up fand frühestens ein Jahr nach der DSE statt und bestand aus einem Telefoninterview oder einer klinischen Untersuchung. Ergebnisse: Die DSE ist zur Diagnose einer TS-LGAS bei Patienten mit einer erhaltenen LVEF von großem Nutzen. Die in der TTE gemessene AVA ist ein unabhängiger Prädiktor für eine TS-LGAS bei Patienten mit erhaltener und reduzierter LVEF. Eine verringerte MAPSE und eine reduzierte TDI-s´ sprechen bei Patienten mit erhaltener LVEF für eine TS-LGAS. Bei Patienten mit reduzierter LVEF weisen ein erhöhter sPAP und eine verringerte AV Geschwindigkeits Ratio auf eine TS-LGAS hin. Bei Zweifeln können weitere bildgebende Verfahren zur Diagnosefindung hinzugezogen werden. N2 - The aim of this study was to determine whether DSE can help differentiate between TS- and PSAS. We also investigated whether there are specific echocardiographic predictors for TS- and PSAS, and whether LVEF plays a crucial role in patients with LGAS. Methods: Both TTE and DSE were performed on 130 patients with asymptomatic AS at the University Hospital Würzburg between January 2011 and December 2016. Various echocardiographic data were collected via TTE, and if the patients had an AVAi ≤ 0.6 cm²/m² and a PGmean < 40 mmHg, they were included in the study. They were divided into two groups depending on whether they had LGAS with an LVEF ≥ 50% or < 50%. In all patients, the AVAproj was calculated during the DSE, and they were then divided into two subgroups: patients with an AVAproj ≤ 1 cm² were assigned to the true-severe LGAS group (TS-LGAS), and patients with an AVAproj > 1 cm² were assigned to the pseudo-severe LGAS group (PS-LGAS). All patient data were manually evaluated. The clinical follow-up took place at least one year after the DSE and consisted of a telephone interview or a clinical examination. Results: DSE is very useful for diagnosing TS-LGAS in patients with preserved LVEF. The AVA measured in TTE is an independent predictor for TS-LGAS in patients with preserved and reduced LVEF. A decreased MAPSE and a reduced TDI-s' indicate TS-LGAS in patients with preserved LVEF. In patients with reduced LVEF, an increased sPAP and a decreased AV velocity ratio indicate TS-LGAS. Additional imaging techniques may be used for diagnosis in cases of doubt. KW - Aortenstenose KW - Dobutamin-Stress-Echokardiographie KW - true-severe aortic stenosis KW - pseudo-severe aortic stenosis Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360146 ER - TY - THES A1 - Buchta, Ulrike T1 - Hörergebnisse und Vestibularisfunktion nach transtemporaler Resektion von intrameatalen Vestibularisschwannomen mit Evaluation der postoperativen, konservativen Hörgeräteversorgung T1 - Hearing results and vestibular function after tumor removal of small vestibular schwannomas by the middle fossa approach and evaluation of the hearing rehabilitation with classical hearing devices postoperatively N2 - Das Vestibularisschwannom ist ein Tumor, dessen Entstehung noch nicht vollständig geklärt ist. Jeder kann von dieser seltenen Erkrankung betroffen sein. Darum ist es wichtig, die Ergebnisse der verschiedenen Therapieoptionen regelmäßig aufzuarbeiten und die Möglichkeiten einer Hörrehabilitation mit konventionellen oder implantierbaren Hörsystemen sowie Ansätze zur Schwindelrehabilitation oder einer möglichen Prähabilitation zu evaluieren. Mit der transtemporalen mikrochirurgischen Tumorexstirpation kann eine Heilung und bei einem großen Teil der Patienten auch ein Hörerhalt erzielt werden. Je jünger die Patienten sind, aber vor allem auch je besser sie vor der Operation hören, desto höher sind auch die Chancen auf einen Hörerhalt. Es zeigte sich jedoch auch, dass bei Patienten mit initial schlechterem Hören, welche dann operiert wurden, dieses zum Teil wieder verbessert werden konnte. Daher sind Ansätze, mehr Patienten eine transtemporale Tumorresektion anzubieten durchaus sinnvoll, auch um die Chancen auf eine Hörverbesserung zu ermöglichen. Des Weiteren sollte auch bei Patienten mit Schwindelbeschwerden eine Operation als Behandlungsoption erwogen werden. Die Prähabilitation mit Gentamicin-Injektionen in das Mittelohr scheint eine neue Möglichkeit zu sein, das zentrale Nervensystem schon auf den Ausfall des Vestibularorgans vorzubereiten. Durch die ototoxische Wirkung des Gentamicins und den potenziellen Hörverlust, wäre jedoch eine Kombination dieser Prähabilitation mit einem hörerhaltenden Eingriff über den transtemporalen Zugang risikobehaftet. Eine zentrale Kompensation der Schwindelbeschwerden sollte nach dem Eingriff mit einer gezielten Schwindelrehabilitation unterstützt werden. Während bei dieser vor allem physiotherapeutische Übungen angewandt werden, gibt es im Hinblick auf eine Hörrehabilitation schon verschiedene medizintechnische Optionen. Aktuell gibt es vor allem Studien zu den implantierbaren Hörhilfen und neue Daten zu den konservativen Möglichkeiten einer Hörrehabilitation sind eher die Ausnahme. Die Nutzung einer konservativen Hörhilfe sollte jedoch vor dem Entscheid zu einer operativen Lösung über implantierbare Systeme konsequent angewandt werden. Die in dieser Arbeit aufgezeigten positiven Momente bei der Versorgung von VS-Patienten prä- und posttherapeutisch mit herkömmlichen Hörgeräten sollten weiteren Eingang in die klinische Routine finden. N2 - The origin of the vestibular schwannoma is not fully understood yet, still anyone can be affected by it. Therefore it´s even more important to compare the different therapeutic options and evaluate the possible option of vestibular and auditory rehabilitation. The tumor removal by the middle fossa approach offers a good therapeutic option with possibility of preserving the natural hearing, especially in younger patients. Some patients even showed an improvement of hearing post-surgery. In patient suffering from vertigo surgery can be an option too. Preoperativly applicated Gentamicin Injections inside the middle ear could be an option to prepare the central nervous system for surgery, on the opposite it acts ototoxic and is not suitable for the goal of a hearing preservation during surgery. While for vestibular rehabilitation physiotherapeutic treatment can be used, there are different ways to accomplish hearing rehabilitation. On one side there are classical hearing aids which should be tried in every case, if these devices are not suiting the needs of the patient implantable hearing aids might be a better option. The results of this study showed a positive outcome for patients treated with both kinds of hearing aids pre- and postoperatively. KW - Akustikustumor KW - Vestibularisschwannom KW - Hörklassifikation KW - transtemporal Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360246 ER - TY - THES A1 - Kühnemundt, Johanna T1 - Defined microphysiologic 3D tumour models with aspects from the tumour microenvironment for the evaluation of cellular immunotherapies T1 - Definierte mikrophysiologische 3D-Tumormodelle mit Aspekten aus der Tumormikroumgebung zur Evaluierung von zellulären Immuntherapien N2 - Adoptive cellular immunotherapy with chimeric antigen receptor (CAR) T cells is highly effective in haematological malignancies. This success, however, has not been achieved in solid tumours so far. In contrast to hematologic malignancies, solid tumours include a hostile tumour microenvironment (TME), that poses additional challenges for curative effects and consistent therapeutic outcome. These challenges manifest in physical and immunological barriers that dampen efficacy of the CAR T cells. Preclinical testing of novel cellular immunotherapies is performed mainly in 2D cell culture and animal experiments. While 2D cell culture is an easy technique for efficacy analysis, animal studies reveal information about toxicity in vivo. However, 2D cell culture cannot fully reflect the complexity observed in vivo, because cells are cultured without anchorage to a matrix and only short-term periods are feasible. Animal studies provide a more complex tissue environment, but xenografts often lack human stroma and tumour inoculation occurs mostly ectopically. This emphasises the need for standardisable and scalable tumour models with incorporated TME-aspects, which enable preclinical testing with enhanced predictive value for the clinical outcome of immunotherapies. Therefore, microphysiologic 3D tumour models based on the biological SISmuc (Small Intestinal mucosa and Submucosa) matrix with preserved basement membrane were engaged and improved in this work to serve as a modular and versatile tumour model for efficacy testing of CAR T cells. In order to reflect a variety of cancer entities, TME-aspects, long-term stability and to enhance the read-out options they were further adapted to achieve scalable and standardisable defined microphysiologic 3D tumour models. In this work, novel culture modalities (semi-static, sandwich-culture) were characterised and established that led to an increased and organised tissue generation and long-term stability. Application of the SISmuc matrix was extended to sarcoma and melanoma models and serial bioluminescence intensity (BLI)-based in vivo imaging analysis was established in the microphysiologic 3D tumour models, which represents a time-efficient read-out method for quality evaluation of the models and treatment efficacy analysis, that is independent of the cell phenotype. Isolation of cancer-associated-fibroblasts (CAFs) from lung (tumour) tissue was demonstrated and CAF-implementation further led to stromal-enriched microphysiologic 3D tumour models with in vivo-comparable tissue-like architecture. Presence of CAFs was confirmed by CAF-associated markers (FAP, α-SMA, MMP-2/-9) and cytokines correlated with CAF phenotype, angiogenesis, invasion and immunomodulation. Additionally, an endothelial cell barrier was implemented for static and dynamic culture in a novel bioreactor set-up, which is of particular interest for the analysis of immune cell diapedesis. Studies in microphysiologic 3D Ewing’s sarcoma models indicated that sarcoma cells could be sensitised for GD2-targeting CAR T cells. After enhancing the scale of assessment of the microphysiologic 3D tumour models and improving them for CAR T cell testing, the tumour models were used to analyse their sensitivity towards differently designed receptor tyrosine kinase-like orphan receptor 1 (ROR1) CAR T cells and to study the effects of the incorporated TME-aspects on the CAR T cell treatment respectively. ROR1 has been described as a suitable target for several malignancies including triple negative breast cancer (TNBC), as well as lung cancer. Therefore, microphysiologic 3D TNBC and lung cancer models were established. Analysis of ROR1 CAR T cells that differed in costimulation, spacer length and targeting domain, revealed, that the microphysiologic 3D tumour models are highly sensitive and can distinguish optimal from sub-optimal CAR design. Here, higher affinity of the targeting domain induced stronger anti-tumour efficacy and anti-tumour function depended on spacer length, respectively. Long-term treatment for 14 days with ROR1 CAR T cells was demonstrated in dynamic microphysiologic 3D lung tumour models, which did not result in complete tumour cell removal, whereas direct injection of CAR T cells into TNBC and lung tumour models represented an alternative route of application in addition to administration via the medium flow, as it induced strong anti-tumour response. Influence of the incorporated TME-aspects on ROR1 CAR T cell therapy represented by CAF-incorporation and/or TGF-β supplementation was analysed. Presence of TGF-β revealed that the specific TGF-β receptor inhibitor SD-208 improves ROR1 CAR T cell function, because it effectively abrogated immunosuppressive effects of TGF-β in TNBC models. Implementation of CAFs should provide a physical and immunological barrier towards ROR1 CAR T cells, which, however, was not confirmed, as ROR1 CAR T cell function was retained in the presence of CAFs in stromal-enriched microphysiologic 3D lung tumour models. The absence of an effect of CAF enrichment on CAR T cell efficacy suggests a missing component for the development of an immunosuppressive TME, even though immunomodulatory cytokines were detected in co-culture models. Finally, improved gene-edited ROR1 CAR T cells lacking exhaustion-associated genes (PD-1, TGF-β-receptor or both) were challenged by the combination of CAF-enrichment and TGF-β in microphysiologic 3D TNBC models. Results indicated that the absence of PD-1 and TGF-β receptor leads to improved CAR T cells, that induce strong tumour cell lysis, and are protected against the hostile TME. Collectively, the microphysiologic 3D tumour models presented in this work reflect aspects of the hostile TME of solid tumours, engage BLI-based analysis and provide long-term tissue homeostasis. Therefore, they present a defined, scalable, reproducible, standardisable and exportable model for translational research with enhanced predictive value for efficacy testing and candidate selection of cellular immunotherapy, as exemplified by ROR1 CAR T cells. N2 - Die adoptive Immuntherapie mit chimären Antigenrezeptor (CAR) exprimierenden T-Zellen zeigt bei hämatologischen Krebsformen eine hohe Wirksamkeit. Bisher konnte dieser Erfolg für solide Tumore nicht erreicht werden. Im Gegensatz zu hämatologischen Krebsformen zeigen solide Tumore eine feindliche Tumormikroumgebung (TME), die zusätzliche Herausforderungen für die Erlangung kurativer Effekte und konsistenter Therapieergebnisse darstellen. Diese Herausforderungen äußern sich in physikalischen und immunologischen Barrieren, welche die Wirksamkeit der CAR-T-Zellen abschwächt. Zur präklinischen Testung neuartiger zellulärer Immuntherapien werden hauptsächlich 2D-Zellkulturen und Tierstudien durchgeführt. 2D-Zellkulturexperimente eignen sich vor allem für Wirksamkeitsanalysen, während Tierstudien Aufschluss über die Toxizität in-vivo geben können. Allerdings kann die 2D-Zellkultur die Komplexität der in-vivo Situation nicht vollständig widerspiegeln, da die Zellen ohne Verankerung an einer Matrix kultiviert werden und nur kurzfristige Zeiträume abgebildet werden können. Tierstudien bieten einen komplexeren Gewebekontext, wobei Xenografts aber oft das humane Stroma fehlt und die Tumorinokulation meist ektopisch erfolgt. Dies unterstreicht den Bedarf an standardisierbaren und skalierbaren Tumormodellen mit inkorporierten TME-Aspekten, die präklinische Testungen mit erhöhtem Vorhersagewert für den klinischen Erfolg von Immuntherapien ermöglichen. Daher wurden in dieser Arbeit mikrophysiologische 3D-Tumormodelle auf Basis der biologischen SISmuc (Small Intestinal mukosa und Submukosa)-Matrix mit erhaltener Basalmembran eingesetzt und verbessert, um als modulares und vielseitiges Tumormodell für die Wirksamkeitsprüfung von CAR T-Zellen zu dienen. Um eine Vielzahl von Krebsentitäten, TME-Aspekte und Langzeitstabilität abzubilden und um die Ausleseparamter zu verbessern, wurden die Tumormodelle weiter angepasst um skalierbare und standardisierbare definierte mikrophysiologische 3D Tumormodelle zu erhalten. In der vorliegenden Arbeit wurden neue Kulturmodalitäten (semistatische Kultur, Sandwich-Kultur) charakterisiert und etabliert, die zu einer vermehrten und erhöhten Gewebebildung sowie Langzeitstabilität der Modelle führen. Die Anwendung der SISmuc-Matrix wurde auf Sarkom- und Melanom-Modelle erweitert und in den mikrophysiologischen 3D-Tumormodellen wurde ein serielles Biolumineszenz-Intensitäts (BLI)-basiertes In-vivo-Analyse-Verfahren etabliert, welches eine zeiteffiziente Methode für die Qualitätsbewertung der Modelle sowie die Analyse der Therapiewirksamkeit darstellt, welche unabhängig vom Zell-Phänotyp ist. Die Isolation von Krebs-assoziierten Fibroblasten (CAFs) aus Lungen-(Tumor) Gewebe wurde demonstriert und die CAF-Implementierung führte des Weiteren zu stromal-angereicherten mikrophysiologischen 3D-Tumormodellen mit in-vivo vergleichbarer gewebeähnlicher Architektur. CAFs wurden mit Hilfe von CAF-assoziierten Markern (FAP, α-SMA, MMP-2/-9) und einer Zytokinanalyse in den Modellen identifiziert. Diese bestätigte ebenfalls Zytokine, welche mit Angiogenese, Invasion und Immunmodulation assoziiert sind. Zusätzlich wurde eine Endothelzellbarriere sowohl in statischer als auch in der dynamischen Kultur implementiert, wofür ein neuer Bioreaktoraufbau verwendet wurde, welcher insbesondere für die Analyse der Immunzelldiapedesis interessant ist. Studien in mikrophysiologischen 3D-Ewing-Sarkom-Modellen zeigten, dass diese für GD2-spezifische CAR-T-Zellen sensibilisiert werden können. Nach der Erweiterung des Untersuchungsumfangs der mikrophysiologischen 3D-Tumormodelle und deren Verbesserung für die CAR-T-Zell-Testung wurden die Tumormodelle verwendet, um ihre Sensitivität gegenüber unterschiedlich designten Rezeptor-Tyrosinkinase-like Orphan-Rezeptor 1 (ROR1) -spezifischen CAR-T-Zellen zu analysieren. Des Weiteren wurden die Auswirkungen der eingebauten TME-Aspekte auf die CAR-T-Therapie untersucht. ROR1 wurde als geeignetes Ziel für verschiedene maligne Erkrankungen beschrieben, darunter auch triple-negtive-breast-cancer (TNBC) und Lungenkrebs. Daher wurden mikrophysiologische 3D-TNBC- und Lungenkrebs-Modelle für die Testungen aufgebaut. Die Analyse von ROR1-CAR-T-Zellen, die sich in Kostimulation, Spacerlänge und der Ziel-Domäne unterschieden, zeigte, dass die mikrophysiologischen 3D-Tumormodelle eine hohe Sensitivität zur Unterscheidung von suboptimal und optimal designten CARs aufweisen. Dabei induzierte eine Ziel-Domäne mit höherer Affinität eine stärkere Anti-Tumor-Wirkung. Zusätzlich war die Anti-Tumor-Funktion abhängig von der Spacerlänge. In dynamischen mikrophysiologischen 3D-Lungentumormodellen wurde eine Langzeitbehandlung über 14 Tage mit ROR1-CAR-T-Zellen realisiert, die jedoch nicht zu einer vollständigen Entfernung der Tumorzellen führte. Die direkte Injektion von CAR-T-Zellen in TNBC- und Lungentumormodellen induzierte eine starke Anti-Tumorantwort und stellt somit neben der Zugabe über den Medienstrom einen alternativen Applikationsweg dar. Des Weiteren wurde der Einfluss der inkorporierten TME-Aspekte auf die ROR1 CAR T-Zelltherapie untersucht, welche sich durch CAF-Inkorporation und/oder TGF-β-Supplementierung darstellten. Die Zugabe von TGF-β zeigte, dass der spezifische TGF-β-Rezeptor-Inhibitor SD-208 die Funktion der ROR1 CAR T-Zellen verbesserte, da er die immunsuppressiven Effekte von TGF-β in TNBC-Modellen effektiv aufhob. Die Implementierung von CAFs sollte eine physikalische und immunologische Barriere gegenüber ROR1 CAR T-Zellen darstellen, was sich jedoch nicht bestätigte, da die Funktion der ROR1 CAR T-Zellen in Anwesenheit von CAFs in stromal-angereicherten mikrophysiologischen 3D-Lungentumormodellen erhalten blieb. Das Fehlen eines Effekts der CAF-Anreicherung auf die CAR T-Zell-Effektivität deutet auf eine fehlende Komponente für die Entwicklung eines immunsuppressiven TME hin, obwohl immunmodulatorische Zytokine in Co-Kultur-Modellen nachgewiesen wurden. Schließlich wurden verbesserte gen-editierte ROR1-CAR-T-Zellen, denen erschöpfungsassoziierte Gene (PD-1, TGF-β-Rezeptor oder beide) fehlten, durch die Kombination von CAF-Anreicherung und TGF-β in mikrophysiologischen 3D-TNBC-Modellen herausgefordert. Die Ergebnisse zeigten, dass ROR1 CAR T Zellen ohne PD-1 und TGF-β-Rezeptor überlegen sind, eine starke Tumorzell-Lyse induzieren und vor der feindlichen TME geschützt sind. Zusammenfassend spiegeln die in dieser Arbeit vorgestellten mikrophysiologischen 3D-Tumormodelle Aspekte der feindlichen TME solider Tumore wider, ermöglichen BLI-basierte Analysen und bieten eine langfristige Gewebehomöostase. Daher stellen sie ein definiertes, skalierbares, reproduzierbares, standardisierbares und exportierbares Modell für die translationale Forschung mit erhöhtem Vorhersagewert dar. Sie können für die Wirksamkeitsprüfung sowie Kandidatenauswahl von zellulären Immuntherapie verwendet werden, was vor allem am Beispiel der ROR1 CAR T-Zellen gezeigt wurde. KW - CAR T cell KW - immunotherapy KW - 3D tumour model KW - solid tumour KW - tumour microenvironment KW - TNBC KW - lung cancer KW - tumour stroma KW - microphysiologic 3D tumour model KW - Immuntherapie KW - Lungenkrebs KW - Stroma KW - Tumormikroumgebung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-276674 ER - TY - JOUR A1 - Gröbner, Susanne N. A1 - Worst, Barbara C. A1 - Weischenfeldt, Joachim A1 - Buchhalter, Ivo A1 - Kleinheinz, Kortine A1 - Rudneva, Vasilisa A. A1 - Johann, Pascal D. A1 - Balasubramanian, Gnana Prakash A1 - Segura-Wang, Maia A1 - Brabetz, Sebastian A1 - Bender, Sebastian A1 - Hutter, Barbara A1 - Sturm, Dominik A1 - Pfaff, Elke A1 - Hübschmann, Daniel A1 - Zipprich, Gideon A1 - Heinold, Michael A1 - Eils, Jürgen A1 - Lawerenz, Christian A1 - Erkek, Serap A1 - Lambo, Sander A1 - Waszak, Sebastian A1 - Blattmann, Claudia A1 - Borkhardt, Arndt A1 - Kuhlen, Michaela A1 - Eggert, Angelika A1 - Fulda, Simone A1 - Gessler, Manfred A1 - Wegert, Jenny A1 - Kappler, Roland A1 - Baumhoer, Daniel A1 - Stefan, Burdach A1 - Kirschner-Schwabe, Renate A1 - Kontny, Udo A1 - Kulozik, Andreas E. A1 - Lohmann, Dietmar A1 - Hettmer, Simone A1 - Eckert, Cornelia A1 - Bielack, Stefan A1 - Nathrath, Michaela A1 - Niemeyer, Charlotte A1 - Richter, Günther H. A1 - Schulte, Johannes A1 - Siebert, Reiner A1 - Westermann, Frank A1 - Molenaar, Jan J. A1 - Vassal, Gilles A1 - Witt, Hendrik A1 - Burkhardt, Birgit A1 - Kratz, Christian P. A1 - Witt, Olaf A1 - van Tilburg, Cornelis M. A1 - Kramm, Christof M. A1 - Fleischhack, Gudrun A1 - Dirksen, Uta A1 - Rutkowski, Stefan A1 - Frühwald, Michael A1 - Hoff, Katja von A1 - Wolf, Stephan A1 - Klingebeil, Thomas A1 - Koscielniak, Ewa A1 - Landgraf, Pablo A1 - Koster, Jan A1 - Resnick, Adam C. A1 - Zhang, Jinghui A1 - Liu, Yanling A1 - Zhou, Xin A1 - Waanders, Angela J. A1 - Zwijnenburg, Danny A. A1 - Raman, Pichai A1 - Brors, Benedikt A1 - Weber, Ursula D. A1 - Northcott, Paul A. A1 - Pajtler, Kristian W. A1 - Kool, Marcel A1 - Piro, Rosario M. A1 - Korbel, Jan O. A1 - Schlesner, Matthias A1 - Eils, Roland A1 - Jones, David T. W. A1 - Lichter, Peter A1 - Chavez, Lukas A1 - Zapatka, Marc A1 - Pfister, Stefan M. T1 - The landscape of genomic alterations across childhood cancers JF - Nature N2 - Pan-cancer analyses that examine commonalities and differences among various cancer types have emerged as a powerful way to obtain novel insights into cancer biology. Here we present a comprehensive analysis of genetic alterations in a pan-cancer cohort including 961 tumours from children, adolescents, and young adults, comprising 24 distinct molecular types of cancer. Using a standardized workflow, we identified marked differences in terms of mutation frequency and significantly mutated genes in comparison to previously analysed adult cancers. Genetic alterations in 149 putative cancer driver genes separate the tumours into two classes: small mutation and structural/copy-number variant (correlating with germline variants). Structural variants, hyperdiploidy, and chromothripsis are linked to TP53 mutation status and mutational signatures. Our data suggest that 7–8% of the children in this cohort carry an unambiguous predisposing germline variant and that nearly 50% of paediatric neoplasms harbour a potentially druggable event, which is highly relevant for the design of future clinical trials. KW - cancer genomics KW - oncogenesis KW - paediatric cancer KW - predictive markers KW - translational research Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229579 VL - 555 ER - TY - JOUR A1 - Harnoš, Jakub A1 - Cañizal, Maria Consuelo Alonso A1 - Jurásek, Miroslav A1 - Kumar, Jitender A1 - Holler, Cornelia A1 - Schambony, Alexandra A1 - Hanáková, Kateřina A1 - Bernatík, Ondřej A1 - Zdráhal, Zbynêk A1 - Gömöryová, Kristína A1 - Gybeľ, Tomáš A1 - Radaszkiewicz, Tomasz Witold A1 - Kravec, Marek A1 - Trantírek, Lukáš A1 - Ryneš, Jan A1 - Dave, Zankruti A1 - Fernández-Llamazares, Ana Iris A1 - Vácha, Robert A1 - Tripsianes, Konstantinos A1 - Hoffmann, Carsten A1 - Bryja, Vítězslav T1 - Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1 JF - Nature Communications N2 - Dishevelled (DVL) is the key component of the Wnt signaling pathway. Currently, DVL conformational dynamics under native conditions is unknown. To overcome this limitation, we develop the Fluorescein Arsenical Hairpin Binder- (FlAsH-) based FRET in vivo approach to study DVL conformation in living cells. Using this single-cell FRET approach, we demonstrate that (i) Wnt ligands induce open DVL conformation, (ii) DVL variants that are predominantly open, show more even subcellular localization and more efficient membrane recruitment by Frizzled (FZD) and (iii) Casein kinase 1 ɛ (CK1ɛ) has a key regulatory function in DVL conformational dynamics. In silico modeling and in vitro biophysical methods explain how CK1ɛ-specific phosphorylation events control DVL conformations via modulation of the PDZ domain and its interaction with DVL C-terminus. In summary, our study describes an experimental tool for DVL conformational sampling in living cells and elucidates the essential regulatory role of CK1ɛ in DVL conformational dynamics. KW - biological techniques KW - cell signalling KW - phosphorylation Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227837 VL - 10 ER - TY - JOUR A1 - Gottschalk, Michael G. A1 - Richter, Jan A1 - Ziegler, Christiane A1 - Schiele, Miriam A. A1 - Mann, Julia A1 - Geiger, Maximilian J. A1 - Schartner, Christoph A1 - Homola, György A. A1 - Alpers, Georg W. A1 - Büchel, Christian A1 - Fehm, Lydia A1 - Fydrich, Thomas A1 - Gerlach, Alexander L. A1 - Gloster, Andrew T. A1 - Helbig-Lang, Sylvia A1 - Kalisch, Raffael A1 - Kircher, Tilo A1 - Lang, Thomas A1 - Lonsdorf, Tina B. A1 - Pané-Farré, Christiane A. A1 - Ströhle, Andreas A1 - Weber, Heike A1 - Zwanzger, Peter A1 - Arolt, Volker A1 - Romanos, Marcel A1 - Wittchen, Hans-Ulrich A1 - Hamm, Alfons A1 - Pauli, Paul A1 - Reif, Andreas A1 - Deckert, Jürgen A1 - Neufang, Susanne A1 - Höfler, Michael A1 - Domschke, Katharina T1 - Orexin in the anxiety spectrum: association of a HCRTR1 polymorphism with panic disorder/agoraphobia, CBT treatment response and fear-related intermediate phenotypes JF - Translational Psychiatry N2 - Preclinical studies point to a pivotal role of the orexin 1 (OX1) receptor in arousal and fear learning and therefore suggest the HCRTR1 gene as a prime candidate in panic disorder (PD) with/without agoraphobia (AG), PD/AG treatment response, and PD/AG-related intermediate phenotypes. Here, a multilevel approach was applied to test the non-synonymous HCRTR1 C/T Ile408Val gene variant (rs2271933) for association with PD/AG in two independent case-control samples (total n = 613 cases, 1839 healthy subjects), as an outcome predictor of a six-weeks exposure-based cognitive behavioral therapy (CBT) in PD/AG patients (n = 189), as well as with respect to agoraphobic cognitions (ACQ) (n = 483 patients, n = 2382 healthy subjects), fMRI alerting network activation in healthy subjects (n = 94), and a behavioral avoidance task in PD/AG pre- and post-CBT (n = 271). The HCRTR1 rs2271933 T allele was associated with PD/AG in both samples independently, and in their meta-analysis (p = 4.2 × 10−7), particularly in the female subsample (p = 9.8 × 10−9). T allele carriers displayed a significantly poorer CBT outcome (e.g., Hamilton anxiety rating scale: p = 7.5 × 10−4). The T allele count was linked to higher ACQ sores in PD/AG and healthy subjects, decreased inferior frontal gyrus and increased locus coeruleus activation in the alerting network. Finally, the T allele count was associated with increased pre-CBT exposure avoidance and autonomic arousal as well as decreased post-CBT improvement. In sum, the present results provide converging evidence for an involvement of HCRTR1 gene variation in the etiology of PD/AG and PD/AG-related traits as well as treatment response to CBT, supporting future therapeutic approaches targeting the orexin-related arousal system. KW - human behaviour KW - molecular neuroscience KW - personalized medicine KW - predictive markers KW - psychiatric disorders Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227479 VL - 9 ER - TY - JOUR A1 - Giampaolo, Sabrina A1 - Wójcik, Gabriela A1 - Klein-Hessling, Stefan A1 - Serfling, Edgar A1 - Patra, Amiya K. T1 - B cell development is critically dependent on NFATc1 activity JF - Cellular & Molecular Immunology N2 - B cell development in bone marrow is a precisely regulated complex process. Through successive stages of differentiation, which are regulated by a multitude of signaling pathways and an array of lineage-specific transcription factors, the common lymphoid progenitors ultimately give rise to mature B cells. Similar to early thymocyte development in the thymus, early B cell development in bone marrow is critically dependent on IL-7 signaling. During this IL-7-dependent stage of differentiation, several transcription factors, such as E2A, EBF1, and Pax5, among others, play indispensable roles in B lineage specification and maintenance. Although recent studies have implicated several other transcription factors in B cell development, the role of NFATc1 in early B cell developmental stages is not known. Here, using multiple gene-manipulated mouse models and applying various experimental methods, we show that NFATc1 activity is vital for early B cell differentiation. Lack of NFATc1 activity in pro-B cells suppresses EBF1 expression, impairs immunoglobulin gene rearrangement, and thereby preBCR formation, resulting in defective B cell development. Overall, deficiency in NFATc1 activity arrested the pro-B cell transition to the pre-B cell stage, leading to severe B cell lymphopenia. Our findings suggest that, along with other transcription factors, NFATc1 is a critical component of the signaling mechanism that facilitates early B cell differentiation. KW - differentiation KW - EBF1 KW - NFATc1 KW - pro-B KW - pre-B Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233006 VL - 16 ER - TY - JOUR A1 - Schwab, Andrea A1 - Meeuwsen, Annick A1 - Ehlicke, Franziska A1 - Hansmann, Jan A1 - Mulder, Lars A1 - Smits, Anthal A1 - Walles, Heike A1 - Kock, Linda T1 - Ex vivo culture platform for assessment of cartilage repair treatment strategies JF - ALTEX - Alternatives to animal experimentation N2 - There is a great need for valuable ex vivo models that allow for assessment of cartilage repair strategies to reduce the high number of animal experiments. In this paper we present three studies with our novel ex vivo osteochondral culture platform. It consists of two separated media compartments for cartilage and bone, which better represents the in vivo situation and enables supply of factors pecific to the different needs of bone and cartilage. We investigated whether separation of the cartilage and bone compartments and/or culture media results in the maintenance of viability, structural and functional properties of cartilage tissue. Next, we valuated for how long we can preserve cartilage matrix stability of osteochondral explants during long-term culture over 84 days. Finally, we determined the optimal defect size that does not show spontaneous self-healing in this culture system. It was demonstrated that separated compartments for cartilage and bone in combination with tissue-specific medium allow for long-term culture of osteochondral explants while maintaining cartilage viability, atrix tissue content, structure and mechanical properties for at least 56 days. Furthermore, we could create critical size cartilage defects of different sizes in the model. The osteochondral model represents a valuable preclinical ex vivo tool for studying clinically relevant cartilage therapies, such as cartilage biomaterials, for their regenerative potential, for evaluation of drug and cell therapies, or to study mechanisms of cartilage regeneration. It will undoubtedly reduce the number of animals needed for in vivotesting. KW - ex vivo model KW - osteochondral biopsy KW - cartilage repair KW - critical size defect KW - replacement Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-181665 VL - 34 IS - 2 ER - TY - THES A1 - Bröhl, Kathleen T1 - „Lanfranks ‚Chirurgia parva‘ in der Abschrift Konrad Schrecks von Aschaffenburg“ als Quelle zur spätmittelalterlich-frühneuzeitlichen Traumatologie T1 - "Lanfrank's 'Chirurgia parva' in the transcript by Konrad Schreck of Aschaffenburg" as a source on late medieval-early modern traumatology N2 - Ziel der vorliegenden Arbeit ist es, „Lanfranks ‚Chirurgia parva‘ in der Abschrift Konrad Schrecks von Aschaffenburg“1 anhand der von Ralf Vollmuth in seiner Habilitationsschrift „Traumatologie und Feldchirurgie an der Wende vom Mittelalter zur Neuzeit“ erarbeiteten Strukturvorgabe inhaltlich zu erschließen. Durch die Aufarbeitung verschiedener chirurgischer Fachbücher und Manuale unter Verwendung einer gemeinsamen Strukturvorlage soll ermöglicht werden, medizinhistorische Quellen kritisch-kontrastiv zu vergleichen. Das bedeutet, dass die Quellen zuerst ediert und anschließend gegebenenfalls übersetzt werden müssen. Im nächsten Schritt werden die verwendeten Arzneimittel – pflanzlicher, tierischer, mineralischer Herkunft – identifiziert und bestimmt. Im Anschluss werden Monographien mit den bestimmenden Inhaltsstoffen und Eigenschaften erstellt. Anhand dieser Pflanzen- und Arzneistoffmonographien, die im Sinne einer Datenbank aufeinander aufbauen, sollte es dann möglich sein, unter modernen pharmakologischen Gesichtspunkten die Wirksamkeit der verwendeten Arzneimittel zu erschließen. Eine ausreichende Zahl von Quellen, die nach einer gemeinsamen Strukturvorlage bearbeitet wurden, kann es schließlich ermöglichen, zu beurteilen, welche der beschriebenen Anwendungen repräsentativ waren, welche Außenseiterstellung einnahmen oder nur theoretische Ansätze bildeten, die praktisch keine Verwendung fanden. N2 - The aim of this dissertation is to analyse the content of "Lanfrank's 'Chirurgia parva' in the transcript by Konrad Schreck of Aschaffenburg " using the structural template developed by Ralf Vollmuth in his habilitation thesis "Traumatologie und Feldchirurgie an der Wende vom Mittelalter zur Neuzeit". By analysing various surgical textbooks and manuals using a common structural template, it should be possible to compare medical-historical sources critically and contrastively. This means that the sources must first be edited and then, if necessary, translated. In the next step, the medicines used - of plant, animal and mineral origin - are identified and determined. Monographs with the determining ingredients and properties are then compiled. On the basis of these plant and drug monographs, which build on each other in the sense of a database, it should then be possible to determine the efficacy of the drugs used from a modern pharmacological point of view. A sufficient number of sources, which have been processed according to a common structural template, can ultimately make it possible to assess which of the applications described were representative, which were outsiders or which were only theoretical approaches that were not used in practice. KW - Konrad Schreck von Aschaffenburg KW - Lanfrancus, Mediolanensis KW - Traumatologie KW - Spätmittelalter KW - Schreck, Konrad KW - Lanfrank, von Mailand Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359227 ER - TY - THES A1 - Kellner [geb. Friedel], Theresa T1 - Suizid durch Selbstverbrennung im Freien - Eine bildmorphologische Analyse der Intensität und Verteilung von Verbrennungen im Zusammenhang mit der Körperposition während des Brandgeschehens T1 - Suicide by self-immolation in the open air - A photo-based analysis of the intensity and distribution of burns in relation to body position during the burn event N2 - Ziel dieser Dissertation ist es, etwaige Gemeinsamkeiten und Unterschiede hinsichtlich der Distribution und Intensität von Brandverletzungen bei suizidaler Selbstverbrennung im Freien in Abhängigkeit von der jeweiligen Körperposition zum Auffindezeitpunkt anhand der Aktenlage herauszuarbeiten. Das Studienkollektiv umfasst 38 Fälle aus 9 deutschen rechtsmedizinischen Instituten, darunter 13 (34,2 %) weibliche und 25 (65,8 %) männliche Suizidenten/-innen im Alter von 18 – 77 Jahren. Neben einer deskriptiven visuellen Analyse erfolgt die Auswertung der Verteilung der Verbrennungen mittels der Software BurnCase 3D, die es ermöglicht, eine Sortierung der einzelnen Körperbereiche nach deren durchschnittlicher Verbrennungsintensität innerhalb verschiedener Cluster für die unterschiedlichen Auffindepositionen am Tatort (Rückenlage, Bauchlage, Seitenlage, Aufrecht, Sitzend) vorzunehmen. Am ehesten auf das in aufrechter Haltung beginnende Brandgeschehen zurückzuführen ist eine clusterübergreifend auftretende, intensive und nach kranial an Intensität abnehmende Verbrennung des Halses sowie der lateralen und perioralen Kopfbereiche. Geringe Verbrennungsfolgen weisen die distalen unteren Extremitäten sowie die Auflageflächen des Körpers auf dem Untergrund auf. Es zeigt sich eine Beeinflussung der lokalen Verbrennungstiefe durch ein hohes Fettgewebevorkommen. Ebenfalls clusterübergreifend können verstärkte Brandwirkungen an den Oberschenkelinnenseiten festgestellt werden. In Rücken- und Bauchlage liegt zudem eine höhere Verbrennungsintensität an den Flanken, den Arminnenseiten und im Unterbauchbereich vor. Bei in Seitenlage verbrannten Körpern ergeben sich Hinweise darauf, dass die nach oben gerichtete Körperseite vermehrt Verbrennungen aufweist. In aufrechter und sitzender Position konzentriert sich der Brandfokus überwiegend auf Torso, Hals und Kopf. Zusätzlich wurde eine Betrachtung des Entstehungsmusters kutaner Hitzerisse durchgeführt. Hier ergaben sich Übereinstimmungen u.a. mit dem Verlauf der Hautfaltlinien nach Pinkus. Ein Körperschema mit Abbildung der beobachteten Orientierungen der Risse wurde angefertigt. Die wichtigsten Limitationen ergeben sich aus einer geringen Fallzahl, einer fotografischen Dokumentation, die nicht alle Körperbereiche in ausreichender Qualität und Detailliertheit abdeckt, sowie dem subjektiven Bias hinsichtlich der Bewertung der Verbrennungsintensitäten. N2 - The aim of this dissertation is to identify any similarities and differences with regard to the distribution and intensity of burn injuries in suicidal self-immolation in the open air depending on the respective body position at the time of discovery on the basis of the records. The study collective comprises 38 cases from 9 German forensic medical institutes, including 13 (34.2 %) female and 25 (65.8 %) male suicides aged between 18 and 77 years. In addition to a descriptive visual analysis, the distribution of burns was evaluated using the BurnCase 3D software, which enables the individual body areas to be sorted according to their average burn intensity within different clusters for the different positions at the crime scene (supine, prone, lateral, upright, sitting). Burns to the neck and the lateral and perioral areas of the head are most likely to be due to the burns beginning in the upright position and occurring across all clusters. The distal lower extremities and the areas where the body rests on the ground show minor burn effects. The local burn depth is influenced by a high occurrence of fatty tissue. Increased burn effects on the inner thighs can also be observed across all clusters. In the supine and prone positions, there is also a higher intensity of burns on the flanks, inner sides of the arms and in the lower abdomen. In the case of bodies burned in the lateral position, there are indications that the upward-facing side of the body shows increased burns. In the upright and sitting position, the focus of the burn is predominantly on the torso, neck and head. In addition, the development pattern of cutaneous heat lacerations was examined. This revealed similarities with the course of the skin fold lines according to Pinkus, among others. A body diagram showing the observed orientations of the lacerations was drawn up. The most important limitations result from a small number of cases, photographic documentation that does not cover all areas of the body in sufficient quality and detail, and the subjective bias with regard to the assessment of burn intensities. KW - Selbstverbrennung KW - Hitzerisse KW - Suizid KW - Verbrennung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359193 ER - TY - THES A1 - Landmesser, Patricia Sophia T1 - Seroprävalenz von SARS-CoV-2 Antikörpern bei Medizinstudierenden im zweiten klinischen Semester von Juli 2020 bis Juni 2021 T1 - Seroprevalence of SARS-CoV-2 antibodies in medical students in the second clinical semester from July 2020 to June 2021 N2 - Im sechsten Semester des Medizinstudiums an der Julius-Maximilians-Universität Würzburg findet das verpflichtende Praktikum „Impfkurs“ statt. Im Rahmen dieses Kurses wurde vom Sommersemester 2020 bis zum Sommersemester 2021 ein standardisierter online Fragebogen erhoben, der unter anderem demographische Daten sowie Expositionsmöglichkeiten gegenüber SARS-CoV-2 im privaten, beruflichen und universitären Umfeld erfragte. Zusätzlich wurde im gleichen Zeitraum der SARS-CoV-2 Serostatus der Medizinstudierenden erhoben und ausgewertet und dieser mit den Daten des Fragebogens zusammengeführt. Dafür wurden Blutproben entnommen, welche im Labor des Instituts für Virologie der Universität Würzburg mittels Western Blot auf IgG/IgM/IgA Antikörper gegen SARS-CoV-2 untersucht wurden. N2 - In the sixth semester of medical studies at the Julius-Maximilians-Universität Würzburg, the compulsory internship “vaccination course” takes place. As part of this course, a standardized online questionnaire was collected from the summer semester 2020 to the summer semester 2021, which, among other things, collected demographic data and exposure to SARS-CoV-2 in the private, professional and university environment. In addition, the SARS-CoV-2 serostatus of the medical students was collected and evaluated during the same period and merged with the data from the questionnaire. For this purpose, blood samples were taken, which were tested for IgG/IgM/IgA antibodies against SARS-CoV-2 by Western blot. KW - SARS-CoV-2 KW - Medizinstudent Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359246 ER - TY - THES A1 - Adam, Pia Sophie T1 - Expression von PD-L1 und FGFR1-4 beim anaplastischen und gering differenzierten Schilddrüsenkarzinom - Evaluation als präklinische diagnostische Marker T1 - FGF-Receptors and PD-L1 in Anaplastic and Poorly Differentiated Thyroid Cancer: Evaluation of the Preclinical Rationale N2 - Background: Treatment options for poorly differentiated (PDTC) and anaplastic (ATC) thyroid carcinoma are unsatisfactory and prognosis is generally poor. Lenvatinib (LEN), a multi-tyrosine kinase inhibitor targeting fibroblast growth factor receptors (FGFR) 1-4 is approved for advanced radioiodine refractory thyroid carcinoma, but response to single agent is poor in ATC. Recent reports of combining LEN with PD-1 inhibitor pembrolizumab (PEM) are promising. Materials and methods: Primary ATC (n=93) and PDTC (n=47) tissue samples diagnosed 1997-2019 at five German tertiary care centers were assessed for PD-L1 expression by immunohistochemistry using Tumor Proportion Score (TPS). FGFR 1-4 mRNA was quantified in 31 ATC and 14 PDTC with RNAscope in-situ hybridization. Normal thyroid tissue (NT) and papillary thyroid carcinoma (PTC) served as controls. Disease specific survival (DSS) was the primary outcome variable. Results: PD-L1 TPS≥50% was observed in 42% of ATC and 26% of PDTC specimens. Mean PD-L1 expression was significantly higher in ATC (TPS 30%) than in PDTC (5%; p<0.01) and NT (0%, p<0.001). 53% of PDTC samples had PD-L1 expression ≤5%. FGFR mRNA expression was generally low in all samples but combined FGFR1-4 expression was significantly higher in PDTC and ATC compared to NT (each p<0.001). No impact of PD-L1 and FGFR 1-4 expression was observed on DSS. Conclusion: High tumoral expression of PD-L1 in a large proportion of ATCs and a subgroup of PDTCs provides a rationale for immune checkpoint inhibition. FGFR expression is low thyroid tumor cells. The clinically observed synergism of PEM with LEN may be caused by immune modulation. N2 - Hintergrund: Die therapeutischen Optionen für das gering differenzierte (PDTC) und anaplastische (ATC) Schilddrüsenkarzinom sind limitiert, weshalb diese Erkrankungen überwiegend mit einer schlechten Prognose einhergehen. Lenvatinib (LEN) ist ein Multityrosinkinase-Inhibitor, der unter anderem die Fibroblasten-Wachstumsfaktor-Rezeptoren (FGFR) 1-4 inhibiert und zur Therapie des fortgeschrittenen radiojodrefraktären Schilddrüsenkarzinoms zugelassen ist. Es zeigt sich nur ein geringes Ansprechen auf die Monotherapie bei ATCs, wobei neuere Studien eine therapeutische Überlegenheit der Kombination aus LEN und dem PD-1-Inhibitor Pembrolizumab (PEM) beschreiben. Material und Methoden: Die Expression von PD-L1 wurde in ATC (n=93)- und PDTC (n=47)-Primärtumorgewebe von 1997-2019 aus fünf deutschen (Universitäts-)Kliniken mittels Immunhistochemie analysiert und mit dem Tumor Proportion Score (TPS) quantifiziert. Der Nachweis von FGFR1-4-mRNA wurde bei 31 ATC- und 14 PDTC-Gewebeproben mittels RNAscope In-situ-Hybridisierung quantifiziert. Als Kontrollgruppe wurde normales Schilddrüsengewebe (NT) und Gewebe von papillären Schilddrüsenkarzinomen (PTC) verwendet. Der primäre Endpunkt war das krankheitsspezifische Überleben (DSS). Ergebnisse: Eine PD-L1-Expression mit einem TPS ≥50% konnte in 42% der ATC- und in 26% der PDTC-Proben nachgewiesen werden. Die mediane PD-L1-Expression war in ATC-(TPS 30%) signifikant höher im Vergleich zu PDTC-Proben (5%; p<0,01) und NT (0%; p<0,001). 53% der PDTC-Proben zeigten eine PD-L1-Expression ≤5%. Die Expression von FGFR-mRNA war in allen Proben sehr gering, wobei die kombinierte FGFR1-4-Expression in PDTC- und ATC-Gewebe im Vergleich zu normalem Schilddrüsengewebe signifikant höher war (jeweils p<0,001). Es ergab sich keine Assoziation zwischen der PD-L1- und FGFR1-4-Expression mit dem krankheitsspezifischen Überleben. Schlussfolgerung: Eine hohe PD-L1-Expression in einem großen Anteil der ATCs und einem Viertel der PDTCs, könnte auf eine Rationale zur Therapieentscheidung für Immuncheckpoint-Inhibioren hinweisen. Die FGFR-Expression war in allen Schilddrüsenkarzinomen sehr gering. Der klinisch beobachtete Synergismus von PEM und LEN könnte durch immunmodulatorische Effekte hervorgerufen werden. KW - Schilddrüsenkrebs KW - Immun-Checkpoint KW - FGFR KW - PD-L1 KW - Immuncheckpointinhibitor KW - Tyrosinkinaseinhibitor KW - Anaplastisches Schilddrüsenkarzinom KW - Gering differenziertes Schilddrüsenkarzinom KW - Protein-Tyrosin-Kinasen KW - Immuntherapie KW - Tyrosinkinase Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359391 ER - TY - JOUR A1 - Belic, Stanislav A1 - Page, Lukas A1 - Lazariotou, Maria A1 - Waaga-Gasser, Ana Maria A1 - Dragan, Mariola A1 - Springer, Jan A1 - Loeffler, Juergen A1 - Morton, Charles Oliver A1 - Einsele, Hermann A1 - Ullmann, Andrew J. A1 - Wurster, Sebastian T1 - Comparative Analysis of Inflammatory Cytokine Release and Alveolar Epithelial Barrier Invasion in a Transwell® Bilayer Model of Mucormycosis JF - Frontiers in Microbiology N2 - Understanding the mechanisms of early invasion and epithelial defense in opportunistic mold infections is crucial for the evaluation of diagnostic biomarkers and novel treatment strategies. Recent studies revealed unique characteristics of the immunopathology of mucormycoses. We therefore adapted an alveolar Transwell® A549/HPAEC bilayer model for the assessment of epithelial barrier integrity and cytokine response to Rhizopus arrhizus, Rhizomucor pusillus, and Cunninghamella bertholletiae. Hyphal penetration of the alveolar barrier was validated by 18S ribosomal DNA detection in the endothelial compartment. Addition of dendritic cells (moDCs) to the alveolar compartment led to reduced fungal invasion and strongly enhanced pro-inflammatory cytokine response, whereas epithelial CCL2 and CCL5 release was reduced. Despite their phenotypic heterogeneity, the studied Mucorales species elicited the release of similar cytokine patterns by epithelial and dendritic cells. There were significantly elevated lactate dehydrogenase concentrations in the alveolar compartment and epithelial barrier permeability for dextran blue of different molecular weights in Mucorales-infected samples compared to Aspergillus fumigatus infection. Addition of monocyte-derived dendritic cells further aggravated LDH release and epithelial barrier permeability, highlighting the influence of the inflammatory response in mucormycosis-associated tissue damage. An important focus of this study was the evaluation of the reproducibility of readout parameters in independent experimental runs. Our results revealed consistently low coefficients of variation for cytokine concentrations and transcriptional levels of cytokine genes and cell integrity markers. As additional means of model validation, we confirmed that our bilayer model captures key principles of Mucorales biology such as accelerated growth in a hyperglycemic or ketoacidotic environment or reduced epithelial barrier invasion upon epithelial growth factor receptor blockade by gefitinib. Our findings indicate that the Transwell® bilayer model provides a reliable and reproducible tool for assessing host response in mucormycosis. KW - mucormycosis KW - alveolar epithelium KW - in vitro model KW - cytokines KW - dendritic cells Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-252477 VL - 9 ER - TY - JOUR A1 - Balasubramanian, Srikkanth A1 - Skaf, Joseph A1 - Holzgrabe, Ulrike A1 - Bharti, Richa A1 - Förstner, Konrad U. A1 - Ziebuhr, Wilma A1 - Humeida, Ute H. A1 - Abdelmohsen, Usama R. A1 - Oelschlaeger, Tobias A. T1 - A new bioactive compound from the marine sponge-derived Streptomyces sp. SBT348 inhibits staphylococcal growth and biofilm formation JF - Frontiers in Microbiology N2 - Staphylococcus epidermidis, the common inhabitant of human skin and mucosal surfaces has emerged as an important pathogen in patients carrying surgical implants and medical devices. Entering the body via surgical sites and colonizing the medical devices through formation of multi-layered biofilms leads to refractory and persistent device-related infections (DRIs). Staphylococci organized in biofilms are more tolerant to antibiotics and immune responses, and thus are difficult-to-treat. The consequent morbidity and mortality, and economic losses in health care systems has strongly necessitated the need for development of new anti-bacterial and anti-biofilm-based therapeutics. In this study, we describe the biological activity of a marine sponge-derived Streptomyces sp. SBT348 extract in restraining staphylococcal growth and biofilm formation on polystyrene, glass, medically relevant titan metal, and silicone surfaces. A bioassay-guided fractionation was performed to isolate the active compound (SKC3) from the crude SBT348 extract. Our results demonstrated that SKC3 effectively inhibits the growth (MIC: 31.25 \(\mu\)g/ml) and biofilm formation (sub-MIC range: 1.95-<31.25 \(\mu\)g/ml) of S. epidermidis RP62A in vitro. Chemical characterization of SKC3 by heat and enzyme treatments, and mass spectrometry (HRMS) revealed its heat-stable and non-proteinaceous nature, and high molecular weight (1258.3 Da). Cytotoxicity profiling of SKC3 in vitro on mouse fibroblast (NIH/3T3) and macrophage (J774.1) cell lines, and in vivo on the greater wax moth larvae Galleria mellonella revealed its non-toxic nature at the effective dose. Transcriptome analysis of SKC3 treated S. epidermidis RP62A has further unmasked its negative effect on central metabolism such as carbon flux as well as, amino acid, lipid, and energy metabolism. Taken together, these findings suggest a potential of SKC3 as a putative drug to prevent staphylococcal DRIs. KW - marine sponges KW - Streptomyces KW - Staphylococci KW - device-related infections KW - bioassay-guided fractionation KW - transcriptome Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221408 VL - 9 ER - TY - JOUR A1 - Joos, J. P. A1 - Saadatmand, A. R. A1 - Schnabel, C. A1 - Viktorinová, I. A1 - Brand, T. A1 - Kramer, M. A1 - Nattel, S. A1 - Dobrev, D. A1 - Tomancak, P. A1 - Backs, J. A1 - Kleinbongard, P. A1 - Heusch, G. A1 - Lorenz, K. A1 - Koch, E. A1 - Weber, S. A1 - El-Armouche, A. T1 - Ectopic expression of S28A-mutated Histone H3 modulates longevity, stress resistance and cardiac function in Drosophila JF - Scientific Reports N2 - Histone H3 serine 28 (H3S28) phosphorylation and de-repression of polycomb repressive complex (PRC)-mediated gene regulation is linked to stress conditions in mitotic and post-mitotic cells. To better understand the role of H3S28 phosphorylation in vivo, we studied a Drosophila strain with ectopic expression of constitutively-activated H3S28A, which prevents PRC2 binding at H3S28, thus mimicking H3S28 phosphorylation. H3S28A mutants showed prolonged life span and improved resistance against starvation and paraquat-induced oxidative stress. Morphological and functional analysis of heart tubes revealed smaller luminal areas and thicker walls accompanied by moderately improved cardiac function after acute stress induction. Whole-exome deep gene-sequencing from isolated heart tubes revealed phenotype-corresponding changes in longevity-promoting and myotropic genes. We also found changes in genes controlling mitochondrial biogenesis and respiration. Analysis of mitochondrial respiration from whole flies revealed improved efficacy of ATP production with reduced electron transport-chain activity. Finally, we analyzed posttranslational modification of H3S28 in an experimental heart failure model and observed increased H3S28 phosphorylation levels in HF hearts. Our data establish a critical role of H3S28 phosphorylation in vivo for life span, stress resistance, cardiac and mitochondrial function in Drosophila. These findings may pave the way for H3S28 phosphorylation as a putative target to treat stress-related disorders such as heart failure. KW - cardiac hypertrophy KW - epigenetics KW - heart failure Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323637 VL - 8 ER - TY - JOUR A1 - Knop, Janin A1 - Spilgies, Lisanne M. A1 - Rufli, Stefanie A1 - Reinhart, Ramona A1 - Vasilikos, Lazaros A1 - Yabal, Monica A1 - Owsley, Erika A1 - Jost, Philipp J. A1 - Marsh, Rebecca A. A1 - Wajant, Harald A1 - Robinson, Mark D. A1 - Kaufmann, Thomas A1 - W. Wei-Lynn, Wong T1 - TNFR2 induced priming of the inflammasome leads to a RIPK1-dependent cell death in the absence of XIAP JF - Cell Death & Disease N2 - The pediatric immune deficiency X-linked proliferative disease-2 (XLP-2) is a unique disease, with patients presenting with either hemophagocytic lymphohistiocytosis (HLH) or intestinal bowel disease (IBD). Interestingly, XLP-2 patients display high levels of IL-18 in the serum even while in stable condition, presumably through spontaneous inflammasome activation. Recent data suggests that LPS stimulation can trigger inflammasome activation through a TNFR2/TNF/TNFR1 mediated loop in xiap−/− macrophages. Yet, the direct role TNFR2-specific activation plays in the absence of XIAP is unknown. We found TNFR2-specific activation leads to cell death in xiap−/− myeloid cells, particularly in the absence of the RING domain. RIPK1 kinase activity downstream of TNFR2 resulted in a TNF/TNFR1 cell death, independent of necroptosis. TNFR2-specific activation leads to a similar inflammatory NF-kB driven transcriptional profile as TNFR1 activation with the exception of upregulation of NLRP3 and caspase-11. Activation and upregulation of the canonical inflammasome upon loss of XIAP was mediated by RIPK1 kinase activity and ROS production. While both the inhibition of RIPK1 kinase activity and ROS production reduced cell death, as well as release of IL-1β, the release of IL-18 was not reduced to basal levels. This study supports targeting TNFR2 specifically to reduce IL-18 release in XLP-2 patients and to reduce priming of the inflammasome components. KW - cell death and immune response KW - inflammation Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325946 VL - 10 ER - TY - JOUR A1 - Kraus, Amelie J. A1 - Brink, Benedikt G. A1 - Siegel, T. Nicolai T1 - Efficient and specific oligo-based depletion of rRNA JF - Scientific Reports N2 - In most organisms, ribosomal RNA (rRNA) contributes to >85% of total RNA. Thus, to obtain useful information from RNA-sequencing (RNA-seq) analyses at reasonable sequencing depth, typically, mature polyadenylated transcripts are enriched or rRNA molecules are depleted. Targeted depletion of rRNA is particularly useful when studying transcripts lacking a poly(A) tail, such as some non-coding RNAs (ncRNAs), most bacterial RNAs and partially degraded or immature transcripts. While several commercially available kits allow effective rRNA depletion, their efficiency relies on a high degree of sequence homology between oligonucleotide probes and the target RNA. This restricts the use of such kits to a limited number of organisms with conserved rRNA sequences. In this study we describe the use of biotinylated oligos and streptavidin-coated paramagnetic beads for the efficient and specific depletion of trypanosomal rRNA. Our approach reduces the levels of the most abundant rRNA transcripts to less than 5% with minimal off-target effects. By adjusting the sequence of the oligonucleotide probes, our approach can be used to deplete rRNAs or other abundant transcripts independent of species. Thus, our protocol provides a useful alternative for rRNA removal where enrichment of polyadenylated transcripts is not an option and commercial kits for rRNA are not available. KW - parasite biology KW - RNA sequencing KW - transcriptomics Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224829 VL - 9 ER - TY - JOUR A1 - Kotz, Frederik A1 - Risch, Patrick A1 - Arnold, Karl A1 - Sevim, Semih A1 - Puigmartí-Luis, Josep A1 - Quick, Alexander A1 - Thiel, Michael A1 - Hrynevich, Andrei A1 - Dalton, Paul D. A1 - Helmer, Dorothea A1 - Rapp, Bastian E. T1 - Fabrication of arbitrary three-dimensional suspended hollow microstructures in transparent fused silica glass JF - Nature Communications N2 - Fused silica glass is the preferred material for applications which require long-term chemical and mechanical stability as well as excellent optical properties. The manufacturing of complex hollow microstructures within transparent fused silica glass is of particular interest for, among others, the miniaturization of chemical synthesis towards more versatile, configurable and environmentally friendly flow-through chemistry as well as high-quality optical waveguides or capillaries. However, microstructuring of such complex three-dimensional structures in glass has proven evasive due to its high thermal and chemical stability as well as mechanical hardness. Here we present an approach for the generation of hollow microstructures in fused silica glass with high precision and freedom of three-dimensional designs. The process combines the concept of sacrificial template replication with a room-temperature molding process for fused silica glass. The fabricated glass chips are versatile tools for, among other, the advance of miniaturization in chemical synthesis on chip. KW - chemical engineering KW - fluidics KW - materials for optics Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224787 VL - 10 ER - TY - JOUR A1 - Hommers, L. G. A1 - Richter, J. A1 - Yang, Y. A1 - Raab, A. A1 - Baumann, C. A1 - Lang, K. A1 - Schiele, M. A. A1 - Weber, H. A1 - Wittmann, A. A1 - Wolf, C. A1 - Alpers, G. W. A1 - Arolt, V. A1 - Domschke, K. A1 - Fehm, L. A1 - Fydrich, T. A1 - Gerlach, A. A1 - Gloster, A. T. A1 - Hamm, A. O. A1 - Helbig-Lang, S. A1 - Kircher, T. A1 - Lang, T. A1 - Pané-Farré, C. A. A1 - Pauli, P. A1 - Pfleiderer, B. A1 - Reif, A. A1 - Romanos, M. A1 - Straube, B. A1 - Ströhle, A. A1 - Wittchen, H.-U. A1 - Frantz, S. A1 - Ertl, G. A1 - Lohse, M. J. A1 - Lueken, U. A1 - Deckert, J. T1 - A functional genetic variation of SLC6A2 repressor hsa-miR-579-3p upregulates sympathetic noradrenergic processes of fear and anxiety JF - Translational Psychiatry N2 - Increased sympathetic noradrenergic signaling is crucially involved in fear and anxiety as defensive states. MicroRNAs regulate dynamic gene expression during synaptic plasticity and genetic variation of microRNAs modulating noradrenaline transporter gene (SLC6A2) expression may thus lead to altered central and peripheral processing of fear and anxiety. In silico prediction of microRNA regulation of SLC6A2 was confirmed by luciferase reporter assays and identified hsa-miR-579-3p as a regulating microRNA. The minor (T)-allele of rs2910931 (MAFcases = 0.431, MAFcontrols = 0.368) upstream of MIR579 was associated with panic disorder in patients (pallelic = 0.004, ncases = 506, ncontrols = 506) and with higher trait anxiety in healthy individuals (pASI = 0.029, pACQ = 0.047, n = 3112). Compared to the major (A)-allele, increased promoter activity was observed in luciferase reporter assays in vitro suggesting more effective MIR579 expression and SLC6A2 repression in vivo (p = 0.041). Healthy individuals carrying at least one (T)-allele showed a brain activation pattern suggesting increased defensive responding and sympathetic noradrenergic activation in midbrain and limbic areas during the extinction of conditioned fear. Panic disorder patients carrying two (T)-alleles showed elevated heart rates in an anxiety-provoking behavioral avoidance test (F(2, 270) = 5.47, p = 0.005). Fine-tuning of noradrenaline homeostasis by a MIR579 genetic variation modulated central and peripheral sympathetic noradrenergic activation during fear processing and anxiety. This study opens new perspectives on the role of microRNAs in the etiopathogenesis of anxiety disorders, particularly their cardiovascular symptoms and comorbidities. KW - clinical genetics KW - psychiatric disorders Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-322497 VL - 8 ER - TY - JOUR A1 - Bury, Susanne A1 - Soundararajan, Manonmani A1 - Bharti, Richa A1 - von Bünau, Rudolf A1 - Förstner, Konrad U. A1 - Oelschlaeger, Tobias A. T1 - The probiotic escherichia coli strain Nissle 1917 combats lambdoid bacteriophages stx and lambda JF - Frontiers in Microbiology N2 - Shiga toxin (Stx) producing E. coli (STEC) such as Enterohemorrhagic E. coli (EHEC) are the major cause of foodborne illness in humans. In vitro studies showed the probiotic Escherichia coil strain Nissle 1917 (EcN) to efficiently inhibit the production of Stx. Life threatening EHEC strains as for example the serotype 0104:H4, responsible for the great outbreak in 2011 in Germany, evolutionary developed from certain E. coll strains which got infected by stx2-encoding lambdoid phages turning the E. coil into lysogenic and subsequently Stx producing strains. Since antibiotics induce stx genes and Stx production, EHEC infected persons are not recommended to be treated with antibiotics. Therefore, EcN might be an alternative medication. However, because even commensal E. coli strains might be converted into Stx-producers after becoming host to a stx encoding prophage, we tested EcN for stx-phage genome integration. Our experiments revealed the resistance of EcN toward not only stx-phages but also against lambda-phages. This resistance was not based on the lack of or by mutated phage receptors. Rather it involved the expression of a phage repressor (pr) gene of a defective prophage in EcN which was able to partially protect E. coli K-12 strain MG1655 against stx and lambda phage infection. Furthermore, we observed EcN to inactivate phages and thereby to protect E. coli K-12 strains against infection by stx- as well as lambda-phages. Inactivation of lambda-phages was due to binding of lambda-phages to LamB of EcN whereas inactivation of stx-phages was caused by a thermostable protein of EcN. These properties together with its ability to inhibit Stx production make EcN a good candidate for the prevention of illness caused by EHEC and probably for the treatment of already infected people. KW - probiotic KW - E. coli Nissle 1917 KW - EHEC KW - Shiga toxin producing E. coli KW - stx-phages KW - lambda-phages KW - lambdoid prophage KW - LamB Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221960 VL - 9 ER - TY - JOUR A1 - Breitinger, Ulrike A1 - Bahnassawy, Lamiaa M. A1 - Janzen, Dieter A1 - Römer, Vera A1 - Becker, Cord-Michael A1 - Villmann, Carmen A1 - Breitinger, Hans-Georg T1 - PKA and PKC modulators affect ion channel function and internalization of recombinant alpha1 and alpha1-beta glycine receptors JF - Frontiers in Molecular Neurosience N2 - Glycine receptors (GlyRs) are important mediators of fast inhibitory neurotransmission in the mammalian central nervous system. Their function is controlled by multiple cellular mechanisms, including intracellular regulatory processes. Modulation of GlyR function by protein kinases has been reported for many cell types, involving different techniques, and often yielding contradictory results. Here, we studied the effects of protein kinase C (PKC) and cAMP-dependent protein kinase A (PKA) on glycine induced currents in HEK293 cells expressing human homomeric \(\alpha\)1 and heteromeric \(\alpha\)1-\(\beta\) GlyRs using whole-cell patch clamp techniques as well as internalization assays. In whole-cell patch-clamp measurements, modulators were applied in the intracellular buffer at concentrations between 0.1 \(\mu\)M and 0.5 \(\mu\)M. EC50 of glycine increased upon application of the protein kinase activators Forskolin and phorbol-12-myristate-13-acetate (PMA) but decreased in the presence of the PKC inhibitor Staurosporine aglycon and the PKA inhibitor H-89. Desensitization of recombinant \(\alpha\)1 receptors was significantly increased in the presence of Forskolin. Staurosporine aglycon, on the other hand decreased desensitization of heteromeric \(\alpha\)1-\(\beta\) GlyRs. The time course of receptor activation was determined for homomeric \(\alpha\)1 receptors and revealed two simultaneous effects: cells showed a decrease of EC50 after 3-6 min of establishing whole-cell configuration. This effect was independent of protein kinase modulators. All modulators of PKA and PKC, however, produced an additional shift of EC50, which overlay and eventually exceeded the cells intrinsic variation of EC50. The effect of kinase activators was abolished if the corresponding inhibitors were co-applied, consistent with PKA and PKC directly mediating the modulation of GlyR function. Direct effects of PKA-and PKC-modulators on receptor expression on transfected HEK cells were monitored within 15 min of drug application, showing a significant increase of receptor internalization with PKA and PKC activators, while the corresponding inhibitors had no significant effect on receptor surface expression or internalization. Our results confirm the observation that phosphorylation via PKA and PKC has a direct effect on the GlyR ion channel complex and plays an important role in the fine-tuning of glycinergic signaling. KW - glycine receptor KW - PKA KW - PKC KW - activators/inhibitors of phosphorylation KW - whole-cell currents KW - modulation kinetics KW - receptor internalization Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220401 VL - 11 ER - TY - JOUR A1 - Bohmann, Ferdinand O. A1 - Kurka, Natalia A1 - du Mesnil de Rochemont, Richard A1 - Gruber, Katharina A1 - Guenther, Joachim A1 - Rostek, Peter A1 - Rai, Heike A1 - Zickler, Philipp A1 - Ertl, Michael A1 - Berlis, Ansgar A1 - Poli, Sven A1 - Mengel, Annerose A1 - Ringleb, Peter A1 - Nagel, Simon A1 - Pfaff, Johannes A1 - Wollenweber, Frank A. A1 - Kellert, Lars A1 - Herzberg, Moriz A1 - Koehler, Luzie A1 - Haeusler, Karl Georg A1 - Alegiani, Anna A1 - Schubert, Charlotte A1 - Brekenfeld, Caspar A1 - Doppler, Christopher E. J. A1 - Onur, Oezguer A. A1 - Kabbasch, Christoph A1 - Manser, Tanja A1 - Pfeilschifter, Waltraud T1 - Simulation-based training of the rapid evaluation and management of acute stroke (STREAM) — a prospective single-arm multicenter trial JF - Frontiers in Neurology N2 - Introduction: Acute stroke care delivered by interdisciplinary teams is time-sensitive. Simulation-based team training is a promising tool to improve team performance in medical operations. It has the potential to improve process times, team communication, patient safety, and staff satisfaction. We aim to assess whether a multi-level approach consisting of a stringent workflow revision based on peer-to-peer review and 2–3 one-day in situ simulation trainings can improve acute stroke care processing times in high volume neurocenters within a 6 months period. Methods and Analysis: The trial is being carried out in a pre-test-post-test design at 7 tertiary care university hospital neurocenters in Germany. The intervention is directed at the interdisciplinary multiprofessional stroke teams. Before and after the intervention, process times of all direct-to-center stroke patients receiving IV thrombolysis (IVT) and/or endovascular therapy (EVT) will be recorded. The primary outcome measure will be the “door-to-needle” time of all consecutive stroke patients directly admitted to the neurocenters who receive IVT. Secondary outcome measures will be intervention-related process times of the fraction of patients undergoing EVT and effects on team communication, perceived patient safety, and staff satisfaction via a staff questionnaire. Interventions: We are applying a multi-level intervention in cooperation with three “STREAM multipliers” from each center. First step is a central meeting of the multipliers at the sponsor's institution with the purposes of algorithm review in a peer-to-peer process that is recorded in a protocol and an introduction to the principles of simulation training and debriefing as well as crew resource management and team communication. Thereafter, the multipliers cooperate with the stroke team trainers from the sponsor's institution to plan and execute 2–3 one-day simulation courses in situ in the emergency department and CT room of the trial centers whereupon they receive teaching materials to perpetuate the trainings. Clinical Trial Registration: STREAM is a registered trial at https://clinicaltrials.gov/ct2/show/NCT03228251. KW - CRM KW - thrombolysis (tPA) KW - stroke KW - emergency care KW - simulation training Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369239 SN - 1664-2295 VL - 10 ER - TY - JOUR A1 - Bolzoni, Francesco A1 - Esposti, Roberto A1 - Marchese, Silvia M. A1 - Pozzi, Nicoló G. A1 - Ramirez-Pasos, Uri E. A1 - Isaias, Ioannis U. A1 - Cavallari, Paolo T1 - Disrupt of intra-limb APA pattern in parkinsonian patients performing index-finger flexion JF - Frontiers in Physiology N2 - Voluntary movements induce postural perturbations which are counteracted by anticipatory postural adjustments (APAs). These actions are known to build up long fixation chains toward available support points (inter-limb APAs), so as to grant whole body equilibrium. Moreover, recent studies highlighted that APAs also build-up short fixation chains, within the same limb where a distal segment is moved (intra-limb APAs), aimed at stabilizing the proximal segments. The neural structures generating intra-limb APAs still need investigations; the present study aims to compare focal movement kinematics and intra-limb APA latencies and pattern between healthy subjects and parkinsonian patients, assuming the latter as a model of basal ganglia dysfunction. Intra-limb APAs that stabilize the arm when the index-finger is briskly flexed were recorded in 13 parkinsonian patients and in 10 age-matched healthy subjects. Index-finger movement was smaller in parkinsonian patients vs. healthy subjects (p = 0.01) and more delayed with respect to the onset of the prime mover flexor digitorum superficialis (FDS, p < 0.0001). In agreement with the literature, in all healthy subjects the FDS activation was preceded by an inhibitory intra-limb APA in biceps brachii (BB) and anterior deltoid (AD), and almost simultaneous to an excitatory intra-limb APA in triceps brachii (TB). In parkinsonian patients, no significant differences were found for TB and AD intra-limb APA timings, however only four patients showed an inhibitory intra-limb APA in BB, while other four did not show any BB intra-limb APAs and five actually developed a BB excitation. The frequency of occurrence of normal sign, lacking, and inverted BB APAs was different in healthy vs. parkinsonian participants (p = 0.0016). The observed alterations in index-finger kinematics and intra-limb APA pattern in parkinsonian patients suggest that basal ganglia, in addition to shaping the focal movement, may also contribute to intra-limb APA control. KW - intra-limb anticipatory postural adjustments KW - Parkinson disease KW - basal ganglia KW - motor control KW - human Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369245 SN - 1664-042X VL - 9 ER - TY - JOUR A1 - Kessler, Almuth F. A1 - Frömbling, Greta E. A1 - Gross, Franziska A1 - Hahn, Mirja A1 - Dzokou, Wilfrid A1 - Ernestus, Ralf-Ingo A1 - Löhr, Mario A1 - Hagemann, Carsten T1 - Effects of tumor treating fields (TTFields) on glioblastoma cells are augmented by mitotic checkpoint inhibition JF - Cell Death Discovery N2 - Tumor treating fields (TTFields) are approved for glioblastoma (GBM) therapy. TTFields disrupt cell division by inhibiting spindle fiber formation. Spindle assembly checkpoint (SAC) inhibition combined with antimitotic drugs synergistically decreases glioma cell growth in cell culture and mice. We hypothesized that SAC inhibition will increase TTFields efficacy. Human GBM cells (U-87 MG, GaMG) were treated with TTFields (200 kHz, 1.7 V/cm) and/or the SAC inhibitor MPS1-IN-3 (IN-3, 4 µM). Cells were counted after 24, 48, and 72 h of treatment and at 24 and 72 h after end of treatment (EOT). Flow cytometry, immunofluorescence microscopy, Annexin-V staining and TUNEL assay were used to detect alterations in cell cycle and apoptosis after 72 h of treatment. The TTFields/IN-3 combination decreased cell proliferation after 72 h compared to either treatment alone (−78.6% vs. TTFields, P = 0.0337; −52.6% vs. IN-3, P = 0.0205), and reduced the number of viable cells (62% less than seeded). There was a significant cell cycle shift from G1 to G2/M phase (P < 0.0001). The apoptotic rate increased to 44% (TTFields 14%, P = 0.0002; IN-3 4%, P < 0.0001). Cell growth recovered 24 h after EOT with TTFields and IN-3 alone, but the combination led to further decrease by 92% at 72 h EOT if IN-3 treatment was continued (P = 0.0288). The combination of TTFields and SAC inhibition led to earlier and prolonged effects that significantly augmented the efficacy of TTFields and highlights a potential new targeted multimodal treatment for GBM. Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325744 VL - 4 ER - TY - JOUR A1 - Kästner, Niklas A1 - Richter, S. Helene A1 - Urbanik, Sarah A1 - Kunert, Joachim A1 - Waider, Jonas A1 - Lesch, Klaus-Peter A1 - Kaiser, Sylvia A1 - Sachser, Norbert T1 - Brain serotonin deficiency affects female aggression JF - Scientific Reports N2 - The neurotransmitter serotonin plays a key role in the control of aggressive behaviour. While so far most studies have investigated variation in serotonin levels, a recently created tryptophan hydroxylase 2 (Tph2) knockout mouse model allows studying effects of complete brain serotonin deficiency. First studies revealed increased aggressiveness in homozygous Tph2 knockout mice in the context of a resident-intruder paradigm. Focussing on females, this study aimed to elucidate effects of serotonin deficiency on aggressive and non-aggressive social behaviours not in a test situation but a natural setting. For this purpose, female Tph2 wildtype (n = 40) and homozygous knockout mice (n = 40) were housed with a same-sex conspecific of either the same or the other genotype in large terraria. The main findings were: knockout females displayed untypically high levels of aggressive behaviour even after several days of co-housing. Notably, in response to aggressive knockout partners, they showed increased levels of defensive behaviours. While most studies on aggression in rodents have focussed on males, this study suggests a significant involvement of serotonin also in the control of female aggression. Future research will show, whether the observed behavioural effects are directly caused by the lack of serotonin or by potential compensatory mechanisms. KW - animal behaviour KW - genetics of the nervous system KW - social behaviour Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325386 VL - 9 ER - TY - JOUR A1 - Hecht, Markus A1 - Meier, Friedegund A1 - Zimmer, Lisa A1 - Polat, Bülent A1 - Loquai, Carmen A1 - Weishaupt, Carsten A1 - Forschner, Andrea A1 - Gutzmer, Ralf A1 - Utikal, Jochen S. A1 - Goldinger, Simone M. A1 - Geier, Michael A1 - Hassel, Jessica C. A1 - Balermpas, Panagiotis A1 - Kiecker, Felix A1 - Rauschenberg, Ricarda A1 - Dietrich, Ursula A1 - Clemens, Patrick A1 - Berking, Carola A1 - Grabenbauer, Gerhard A1 - Schadendorf, Dirk A1 - Grabbe, Stephan A1 - Schuler, Gerold A1 - Fietkau, Rainer A1 - Distel, Luitpold V. A1 - Heinzerling, Lucie T1 - Clinical outcome of concomitant vs interrupted BRAF inhibitor therapy during radiotherapy in melanoma patients JF - British Journal of Cancer N2 - Background: Concomitant radiation with BRAF inhibitor (BRAFi) therapy may increase radiation-induced side effects but also potentially improve tumour control in melanoma patients. Methods: A total of 155 patients with BRAF-mutated melanoma from 17 European skin cancer centres were retrospectively analysed. Out of these, 87 patients received concomitant radiotherapy and BRAFi (59 vemurafenib, 28 dabrafenib), while in 68 patients BRAFi therapy was interrupted during radiation (51 vemurafenib, 17 dabrafenib). Overall survival was calculated from the first radiation (OSRT) and from start of BRAFi therapy (OSBRAFi). Results: The median duration of BRAFi treatment interruption prior to radiotherapy was 4 days and lasted for 17 days. Median OSRT and OSBRAFi in the entire cohort were 9.8 and 12.6 months in the interrupted group and 7.3 and 11.5 months in the concomitant group (P=0.075/P=0.217), respectively. Interrupted vemurafenib treatment with a median OSRT and OSBRAFi of 10.1 and 13.1 months, respectively, was superior to concomitant vemurafenib treatment with a median OSRT and OSBRAFi of 6.6 and 10.9 months (P=0.004/P=0.067). Interrupted dabrafenib treatment with a median OSRT and OSBRAFi of 7.7 and 9.8 months, respectively, did not differ from concomitant dabrafenib treatment with a median OSRT and OSBRAFi of 9.9 and 11.6 months (P=0.132/P=0.404). Median local control of the irradiated area did not differ in the interrupted and concomitant BRAFi treatment groups (P=0.619). Skin toxicity of grade ≥2 (CTCAE) was significantly increased in patients with concomitant vemurafenib compared to the group with treatment interruption (P=0.002). Conclusions: Interruption of vemurafenib treatment during radiation was associated with better survival and less toxicity compared to concomitant treatment. Due to lower number of patients, the relevance of treatment interruption in dabrafenib treated patients should be further investigated. The results of this analysis indicate that treatment with the BRAFi vemurafenib should be interrupted during radiotherapy. Prospective studies are desperately needed. KW - radiation KW - radiotherapy KW - BRAF KW - vemurafenib KW - dabrafenib Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227970 VL - 118 ER - TY - JOUR A1 - Knödler, Maren A1 - Körfer, Justus A1 - Kunzmann, Volker A1 - Trojan, Jörg A1 - Daum, Severin A1 - Schenk, Michael A1 - Kullmann, Frank A1 - Schroll, Sebastian A1 - Behringer, Dirk A1 - Stahl, Michael A1 - Al-Batran, Salah-Eddin A1 - Hacker, Ulrich A1 - Ibach, Stefan A1 - Lindhofer, Horst A1 - Lordick, Florian T1 - Randomised phase II trial to investigate catumaxomab (anti-EpCAM × anti-CD3) for treatment of peritoneal carcinomatosis in patients with gastric cancer JF - British Journal of Cancer N2 - Background Peritoneal carcinomatosis (PC) represents an unfavourable prognostic factor for patients with gastric cancer (GC). Intraperitoneal treatment with the bispecific and trifunctional antibody catumaxomab (EpCAM, CD3), in addition to systemic chemotherapy, could improve elimination of PC. Methods This prospective, randomised, phase II study investigated the efficacy of catumaxomab followed by chemotherapy (arm A, 5-fluorouracil, leucovorin, oxaliplatin, docetaxel, FLOT) or FLOT alone (arm B) in patients with GC and PC. Primary endpoint was the rate of macroscopic complete remission (mCR) of PC at the time of second diagnostic laparoscopy/laparotomy prior to optional surgery. Results Median follow-up was 52 months. Out of 35 patients screened, 15 were allocated to arm A and 16 to arm B. mCR rate was 27% in arm A and 19% in arm B (p = 0.69). Severe side effects associated with catumaxomab were nausea, infection, abdominal pain, and elevated liver enzymes. Median progression-free (6.7 vs. 5.4 months, p = 0.71) and overall survival (13.2 vs. 13.0 months, p = 0.97) were not significantly different in both treatment arms. Conclusions Addition of catumaxomab to systemic chemotherapy was feasible and tolerable in advanced GC. Although the primary endpoint could not be demonstrated, results are promising for future investigations integrating intraperitoneal immunotherapy into a multimodal treatment strategy. KW - cancer immunotherapy KW - gastric cancer Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325938 VL - 119 ER - TY - JOUR A1 - Altieri, Barbara A1 - Di Dato, Carla A1 - Martini, Chiara A1 - Sciammarella, Concetta A1 - Di Sarno, Antonella A1 - Colao, Annamaria A1 - Faggiano, Antongiulio T1 - Bone Metastases in Neuroendocrine Neoplasms: From Pathogenesis to Clinical Management JF - Cancers N2 - Bone represents a common site of metastases for several solid tumors. However, the ability of neuroendocrine neoplasms (NENs) to localize to bone has always been considered a rare and late event. Thanks to the improvement of therapeutic options, which results in longer survival, and of imaging techniques, particularly after the introduction of positron emission tomography (PET) with gallium peptides, the diagnosis of bone metastases (BMs) in NENs is increasing. The onset of BMs can be associated with severe skeletal complications that impair the patient's quality of life. Moreover, BMs negatively affect the prognosis of NEN patients, bringing out the lack of curative treatment options for advanced NENs. The current knowledge on BMs in gastro-entero-pancreatic (GEP) and bronchopulmonary (BP) NENs is still scant and is derived from a few retrospective studies and case reports. This review aims to perform a critical analysis of the evidence regarding the role of BMs in GEP- and BP-NENs, focusing on the molecular mechanisms underlining the development of BMs, as well as clinical presentation, diagnosis, and treatment of BMs, in an attempt to provide suggestions that can be used in clinical practice. KW - neuroendocrine neoplasms KW - bone metastases KW - bone microenvironment KW - skeletal-related events KW - epithelial-to-mesenchymal transition KW - microRNA KW - prognosis KW - treatment KW - denosumab Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221079 VL - 11 ER - TY - JOUR A1 - El-Helou, Sabine M. A1 - Biegner, Anika-Kerstin A1 - Bode, Sebastian A1 - Ehl, Stephan R. A1 - Heeg, Maximilian A1 - Maccari, Maria E. A1 - Ritterbusch, Henrike A1 - Speckmann, Carsten A1 - Rusch, Stephan A1 - Scheible, Raphael A1 - Warnatz, Klaus A1 - Atschekzei, Faranaz A1 - Beider, Renata A1 - Ernst, Diana A1 - Gerschmann, Stev A1 - Jablonka, Alexandra A1 - Mielke, Gudrun A1 - Schmidt, Reinhold E. A1 - Schürmann, Gesine A1 - Sogkas, Georgios A1 - Baumann, Ulrich H. A1 - Klemann, Christian A1 - Viemann, Dorothee A1 - Bernuth, Horst von A1 - Krüger, Renate A1 - Hanitsch, Leif G. A1 - Scheibenbogen, Carmen M. A1 - Wittke, Kirsten A1 - Albert, Michael H. A1 - Eichinger, Anna A1 - Hauck, Fabian A1 - Klein, Christoph A1 - Rack-Hoch, Anita A1 - Sollinger, Franz M. A1 - Avila, Anne A1 - Borte, Michael A1 - Borte, Stephan A1 - Fasshauer, Maria A1 - Hauenherm, Anja A1 - Kellner, Nils A1 - Müller, Anna H. A1 - Ülzen, Anett A1 - Bader, Peter A1 - Bakhtiar, Shahrzad A1 - Lee, Jae-Yun A1 - Heß, Ursula A1 - Schubert, Ralf A1 - Wölke, Sandra A1 - Zielen, Stefan A1 - Ghosh, Sujal A1 - Laws, Hans-Juergen A1 - Neubert, Jennifer A1 - Oommen, Prasad T. A1 - Hönig, Manfred A1 - Schulz, Ansgar A1 - Steinmann, Sandra A1 - Klaus, Schwarz A1 - Dückers, Gregor A1 - Lamers, Beate A1 - Langemeyer, Vanessa A1 - Niehues, Tim A1 - Shai, Sonu A1 - Graf, Dagmar A1 - Müglich, Carmen A1 - Schmalzing, Marc T. A1 - Schwaneck, Eva C. A1 - Tony, Hans-Peter A1 - Dirks, Johannes A1 - Haase, Gabriele A1 - Liese, Johannes G. A1 - Morbach, Henner A1 - Foell, Dirk A1 - Hellige, Antje A1 - Wittkowski, Helmut A1 - Masjosthusmann, Katja A1 - Mohr, Michael A1 - Geberzahn, Linda A1 - Hedrich, Christian M. A1 - Müller, Christiane A1 - Rösen-Wolff, Angela A1 - Roesler, Joachim A1 - Zimmermann, Antje A1 - Behrends, Uta A1 - Rieber, Nikolaus A1 - Schauer, Uwe A1 - Handgretinger, Rupert A1 - Holzer, Ursula A1 - Henes, Jörg A1 - Kanz, Lothar A1 - Boesecke, Christoph A1 - Rockstroh, Jürgen K. A1 - Schwarze-Zander, Carolynne A1 - Wasmuth, Jan-Christian A1 - Dilloo, Dagmar A1 - Hülsmann, Brigitte A1 - Schönberger, Stefan A1 - Schreiber, Stefan A1 - Zeuner, Rainald A1 - Ankermann, Tobias A1 - Bismarck, Philipp von A1 - Huppertz, Hans-Iko A1 - Kaiser-Labusch, Petra A1 - Greil, Johann A1 - Jakoby, Donate A1 - Kulozik, Andreas E. A1 - Metzler, Markus A1 - Naumann-Bartsch, Nora A1 - Sobik, Bettina A1 - Graf, Norbert A1 - Heine, Sabine A1 - Kobbe, Robin A1 - Lehmberg, Kai A1 - Müller, Ingo A1 - Herrmann, Friedrich A1 - Horneff, Gerd A1 - Klein, Ariane A1 - Peitz, Joachim A1 - Schmidt, Nadine A1 - Bielack, Stefan A1 - Groß-Wieltsch, Ute A1 - Classen, Carl F. A1 - Klasen, Jessica A1 - Deutz, Peter A1 - Kamitz, Dirk A1 - Lassy, Lisa A1 - Tenbrock, Klaus A1 - Wagner, Norbert A1 - Bernbeck, Benedikt A1 - Brummel, Bastian A1 - Lara-Villacanas, Eusebia A1 - Münstermann, Esther A1 - Schneider, Dominik T. A1 - Tietsch, Nadine A1 - Westkemper, Marco A1 - Weiß, Michael A1 - Kramm, Christof A1 - Kühnle, Ingrid A1 - Kullmann, Silke A1 - Girschick, Hermann A1 - Specker, Christof A1 - Vinnemeier-Laubenthal, Elisabeth A1 - Haenicke, Henriette A1 - Schulz, Claudia A1 - Schweigerer, Lothar A1 - Müller, Thomas G. A1 - Stiefel, Martina A1 - Belohradsky, Bernd H. A1 - Soetedjo, Veronika A1 - Kindle, Gerhard A1 - Grimbacher, Bodo T1 - The German national registry of primary immunodeficiencies (2012-2017) JF - Frontiers in Immunology N2 - Introduction: The German PID-NET registry was founded in 2009, serving as the first national registry of patients with primary immunodeficiencies (PID) in Germany. It is part of the European Society for Immunodeficiencies (ESID) registry. The primary purpose of the registry is to gather data on the epidemiology, diagnostic delay, diagnosis, and treatment of PIDs. Methods: Clinical and laboratory data was collected from 2,453 patients from 36 German PID centres in an online registry. Data was analysed with the software Stata® and Excel. Results: The minimum prevalence of PID in Germany is 2.72 per 100,000 inhabitants. Among patients aged 1-25, there was a clear predominance of males. The median age of living patients ranged between 7 and 40 years, depending on the respective PID. Predominantly antibody disorders were the most prevalent group with 57% of all 2,453 PID patients (including 728 CVID patients). A gene defect was identified in 36% of patients. Familial cases were observed in 21% of patients. The age of onset for presenting symptoms ranged from birth to late adulthood (range 0-88 years). Presenting symptoms comprised infections (74%) and immune dysregulation (22%). Ninety-three patients were diagnosed without prior clinical symptoms. Regarding the general and clinical diagnostic delay, no PID had undergone a slight decrease within the last decade. However, both, SCID and hyper IgE-syndrome showed a substantial improvement in shortening the time between onset of symptoms and genetic diagnosis. Regarding treatment, 49% of all patients received immunoglobulin G (IgG) substitution (70%-subcutaneous; 29%-intravenous; 1%-unknown). Three-hundred patients underwent at least one hematopoietic stem cell transplantation (HSCT). Five patients had gene therapy. Conclusion: The German PID-NET registry is a precious tool for physicians, researchers, the pharmaceutical industry, politicians, and ultimately the patients, for whom the outcomes will eventually lead to a more timely diagnosis and better treatment. KW - registry for primary immunodeficiency KW - primary immunodeficiency (PID) KW - German PID-NET registry KW - PID prevalence KW - European Society for Immunodeficiencies (ESID) KW - IgG substitution therapy KW - CVID Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226629 VL - 10 ER - TY - JOUR A1 - Butt, Elke A1 - Raman, Dayanidhi T1 - New frontiers for the cytoskeletal protein LASP1 JF - Frontiers in Oncology N2 - In the recent two decades, LIM and SH3 protein 1 (LASP1) has been developed from a simple actin-binding structural protein to a tumor biomarker and subsequently to a complex, nuclear transcriptional regulator. Starting with a brief historical perspective, this review will mainly compare and contrast LASP1 and LASP2 from the angle of the newest data and importantly, examine their role in transcriptional regulation. We will summarize the current knowledge through pictorial models and tables including the roles of different microRNAs in the differential regulation of LASP1 levels and patient outcome rather than specify in detail all tumor entities. Finally, the novel functional roles of LASP1 in secretion of vesicles, expression of matrix metalloproteinases and transcriptional regulation as well as the activation of survival and proliferation pathways in different cancer types are described. KW - LASP1 KW - LASP2 KW - transcriptional regulation KW - nuclear role KW - matrix metalloproteinases KW - AP1 Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221975 VL - 8 ER - TY - JOUR A1 - Doppler, Christopher E. J. A1 - Meyer, Linda A1 - Dovern, Anna A1 - Stühmer-Beckh, Jaro A1 - Weiss, Peter H. A1 - Fink, Gereon R. T1 - Differential impact of social and monetary reward on procedural learning and consolidation in aging and its structural correlates JF - Frontiers in Aging Neuroscience N2 - In young (n = 36, mean +/- SD: 24.8 +/- 4.5 years) and older (n = 34, mean +/- SD: 65.1 +/- 6.5 years) healthy participants, we employed a modified version of the Serial Reaction Time task to measure procedural learning (PL) and consolidation while providing monetary and social reward. Using voxel-based morphometry (VBM), we additionally determined the structural correlates of reward-related motor performance (RMP) and PL. Monetary reward had a beneficial effect on PL in the older subjects only. In contrast, social reward significantly enhanced PL in the older and consolidation in the young participants. VBM analyses revealed that motor performance related to monetary reward was associated with larger grey matter volume (GMV) of the left striatum in the young, and motor performance related to social reward with larger GMV of the medial orbitofrontal cortex in the older group. The differential effects of social reward in young (improved consolidation) and both social and monetary rewards in older (enhanced PL) healthy subjects point to the potential of rewards for interventions targeting aging-associated motor decline or stroke-induced motor deficits. KW - serial reaction time task KW - procedural learning KW - reinforcement learning KW - voxel-based morphometry KW - motor aging Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222394 VL - 11 ER - TY - JOUR A1 - Doppler, Kathrin A1 - Brockmann, Kathrin A1 - Sedghi, Annahita A1 - Wurster, Isabel A1 - Volkmann, Jens A1 - Oertel, Wolfgang H. A1 - Sommer, Claudia T1 - Dermal phospho-alpha-synuclein deposition in patients with Parkinson's disease and mutation of the glucocerebrosidase gene JF - Frontiers in Neurology N2 - Heterozygous mutations in the glucocerebrosidase gene (GBA1) represent the most common genetic risk factor for Parkinson's disease (PD) and are histopathologically associated with a widespread load of alpha-synuclein in the brain. Therefore, PD patients with GBA1 mutations are a cohort of high interest for clinical trials on disease-modifying therapies targeting alpha-synuclein. There is evidence that detection of phospho-alpha-synuclein (p-syn) in dermal nerve fibers might be a biomarker for the histopathological identification of PD patients even at premotor or very early stages of disease. It is so far unknown whether dermal p-syn deposition can also be found in PD patients with GBA1 mutations and may serve as a biomarker for PD in these patients. Skin biopsies of 10 PD patients with different GBA1 mutations (six N3705, three E326K, one L444P) were analyzed by double-immunofluorescence labeling with anti-p-syn and anti-protein gene product 9.5 (PGP9.5, axonal marker) to detect intraaxonal p-syn deposition. Four biopsy sites (distal, proximal leg, paravertebral Th10, and C7) per patient were studied. P-syn was found in six patients (three N370S, three E326K). P-syn deposition was mainly detected in autonomic nerve fibers, but also in somatosensory fibers and was not restricted to a certain GBA1 mutation. In summary, dermal p-syn in PD patients with GBA1 mutations seems to offer a similar distribution and frequency as observed in patients without a known mutation. Skin biopsy may be suitable to study p-syn deposition in these patients or even to identify premotor patients with GBA1 mutations. KW - Parkinson's disease KW - glucocerebrosidase mutation KW - alpha-synuclein KW - skin biopsy KW - biomarker Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222828 VL - 9 ER - TY - THES A1 - Kemmer, Luisa Diana T1 - Darstellung von Inflammation in Atherosklerose mit dem CXCR4-gerichteten PET-Tracer \(^{68}\)Ga-Pentixafor im Vergleich zur \(^{18}\)F-FDG-PET/CT T1 - Imaging inflammation in atherosclerosis with the CXCR4-targeted PET tracer \(^{68}\)Ga-Pentixafor compared to \(^{18}\)F-FDG-PET/CT N2 - Herz-Kreislauf-Erkrankungen zählen zu den häufigsten Todesursachen weltweit. Ein ihr zugrundeliegender pathologischer Prozess ist die Atherosklerose. Die Ruptur eines atheroskelrotischen Plaques kann lebensbedrohlich sein. Derzeit existieren weder ein evaluierter Biomarker noch eine Bildgebungstechnik, die das Risiko einer solchen Plaqueruptur und eines nachfolgenden akuten kardiovaskulären Ereignisses vorhersagen können. Um die bildgebenden Verfahren zur Detektion der Atherosklerose zu optimieren, wurde in dieser Dissertationsarbeit untersucht, ob der PET/CT-Tracer 68Ga-Pentixafor im Vergleich zu 18F-FDG bessere Ergebnisse in der Diagnostik der Atherosklerose erzielen kann. Insgesamt wurden 144 onkologische Patienten in die Studie einbezogen, bei denen die 18F-FDG-PET/CT sowie 68Ga-Pentifaxor-PET/CT aus klinischen Gründen indiziert waren. Befunde, bei denen eine gegenüber dem Hintergrund vermehrte Speicherung ohne physiologische Erklärung nachgewiesen werden konnte, wurden als positiv bewertet. Um Unterschiede zwischen den Patienten außer Acht lassen zu können, wurde die target-to-background-ratio (TBR) berechnet. Zur Beschreibung der Speicherintensität einer Läsion wurde der standardized uptake value (SUV) bestimmt. Nach Auswertung der Daten zeigte sich eine mäßige Korrelation der Anzahl von 68Ga-Pentixafor-positiven Läsionen mit der Anzahl der 18F-FDG positiven Läsionen. Die CXCR4-gerichtete Bildgebung mit 68Ga-Pentixafor identifizierte mehr Läsionen als die 18F-FDG-PET/CT. Bezüglich ihres Verteilungsmusters wiesen die beiden Tracer eine geringe Korrelation auf. Die Aufnahmeintensität beider Tracer korrelierte umgekehrt mit dem Ausmaß der Verkalkung. Stark verkalkte Plaques zeigten die niedrigste Traceraufnahme für beide PET-Tracer. Weitere Studien zur Aufklärung der zugrunde liegenden biologischen Mechanismen und Quellen der CXCR4-Positivität sind in hohem Maße gerechtfertigt. N2 - Cardiovascular diseases are among the most common causes of death worldwide. A pathological process underlying these diseases is atherosclerosis. The rupture of an atherosclerotic plaque can be life-threatening. Currently, there is neither an evaluated biomarker nor an imaging technique that can predict the risk of such a plaque rupture and subsequent acute cardiovascular event. To optimize imaging methods for the detection of atherosclerosis, this dissertation investigated whether the PET/CT tracer 68Ga-Pentixafor can achieve better diagnostic results for atherosclerosis compared to 18F-FDG. A total of 144 oncological patients were included in the study, for whom 18F-FDG-PET/CT and 68Ga-Pentixafor-PET/CT were clinically indicated. Lesions showing increased uptake compared to the background without physiological explanation were rated as positive. To disregard differences between patients, the target-to-background ratio (TBR) was calculated. To describe the uptake intensity of a lesion, the standardized uptake value (SUV) was determined. After evaluating the data, a moderate correlation was observed between the number of 68Ga-Pentixafor-positive lesions and the number of 18F-FDG-positive lesions. CXCR4-targeted imaging with 68Ga-Pentixafor identified more lesions than 18F-FDG-PET/CT. Regarding their distribution patterns, the two tracers showed a low correlation. The uptake intensity of both tracers inversely correlated with the extent of calcification. Highly calcified plaques exhibited the lowest tracer uptake for both PET tracers. Further studies to elucidate the underlying biological mechanisms and sources of CXCR4 positivity are highly warranted. KW - Arteriosklerose KW - Positronen-Emissions-Tomografie KW - Chemokinrezpetor KW - CXCR4 KW - \(^{68}\)Ga-Pentixafor KW - \(^{18}\)F-FDG Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360013 ER - TY - THES A1 - Zillig, Anna-Lena Christina T1 - Einfluss von Sicherheit auf die Schmerzverarbeitung T1 - The influence of safety on pain perception N2 - Im Rahmen des interdisziplinären Promotionsschwerpunkts Resilienzfaktoren der Schmerzverarbeitung des evangelischen Studienwerks in Zusammenarbeit mit der Julius-Maximilians-Universität Würzburg und der Otto-Friedrich-Universität Bamberg untersuche ich in diesem Promotionsprojekt den Einfluss von Sicherheit auf die Schmerzverarbeitung. Es ist bekannt, dass die Schmerzverarbeitung durch Emotionen moduliert werden kann. Man geht davon aus, dass negative Emotionen den Schmerz in der Regel verstärken, während positive Emotionen zu einer Schmerzreduktion führen. Frühere Studien fanden heraus, dass die Erwartung eines aversiven Ereignisses zu Bedrohung und stärkeren Schmerzen führt. Es stellt sich die Frage, ob das Gegenteil von Bedrohung, nämlich Sicherheit, zu einer Verringerung der Schmerzen führen kann. Um diese Hypothese zu untersuchen, habe ich drei Experimente an gesunden ProbandInnen durchgeführt. N2 - I am investigating the influence of safety on pain processing in the present dissertation project as part of the interdisciplinary doctoral program on resilience factors of pain processing of the Evangelisches Studienwerk in cooperation with the Julius-Maximilians-University of Würzburg and the Otto-Friedrich-University of Bamberg. It is known that pain processing is susceptible to an individual’s emotional state, such that negative emotions mostly increase pain while positive emotions lead to a pain decrease. Previous studies found that the anticipation of an aversive event induces threat and elevated pain. The question arises whether the exact opposite, namely safety, can lead to a reduction in pain. To investigate this hypothesis, I conducted three experiments in healthy volunteers. KW - Sicherheit KW - Schmerzverarbeitung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359282 ER - TY - JOUR A1 - Vural, Atay A1 - Doppler, Kathrin A1 - Meinl, Edgar T1 - Autoantibodies Against the Node of Ranvier in Seropositive Chronic Inflammatory Demyelinating Polyneuropathy: Diagnostic, Pathogenic, and Therapeutic Relevance JF - Frontiers in Immunology N2 - Discovery of disease-associated autoantibodies has transformed the clinical management of a variety of neurological disorders. Detection of autoantibodies aids diagnosis and allows patient stratification resulting in treatment optimization. In the last years, a set of autoantibodies against proteins located at the node of Ranvier has been identified in patients with chronic inflammatory demyelinating polyneuropathy (CIDP). These antibodies target neurofascin, contactin1, or contactin-associated protein 1, and we propose to name CIDP patients with these antibodies collectively as seropositive. They have unique clinical characteristics that differ from seronegative CIDP. Moreover, there is compelling evidence that autoantibodies are relevant for the pathogenesis. In this article, we review the current knowledge on the characteristics of autoantibodies against the node of Ranvier proteins and their clinical relevance in CIDP. We start with a description of the structure of the node of Ranvier followed by a summary of assays used to identify seropositive patients; and then, we describe clinical features and characteristics linked to seropositivity. We review knowledge on the role of these autoantibodies for the pathogenesis with relevance for the emerging concept of nodopathy/paranodopathy and summarize the treatment implications. KW - autoantibody KW - seropositive KW - chronic inflammatory demyelinating polyneuropathy KW - node of Ranvier KW - paranode KW - neurofascin KW - contactin KW - contactin-associated protein 1 Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233279 VL - 9 ER - TY - JOUR A1 - Siebert, Claudia A1 - Ciato, Denis A1 - Murakami, Masanori A1 - Frei-Stuber, Ludwig A1 - Perez-Rivas, Luis Gustavo A1 - Monteserin-Garcia, José Luis A1 - Nölting, Svenja A1 - Maurer, Julian A1 - Feuchtinger, Annette A1 - Walch, Axel K. A1 - Haak, Harm R. A1 - Bertherat, Jérôme A1 - Mannelli, Massimo A1 - Fassnacht, Martin A1 - Korpershoek, Esther A1 - Reincke, Martin A1 - Stalla, Günter K. A1 - Hantel, Constanze A1 - Beuschlein, Felix T1 - Heat Shock Protein 90 as a Prognostic Marker and Therapeutic Target for Adrenocortical Carcinoma JF - Frontiers in Endocrinology N2 - Background: Adrenocortical carcinoma (ACC) is a rare tumor entity with restricted therapeutic opportunities. HSP90 (Heat Shock Protein 90) chaperone activity is fundamental for cell survival and contributes to different oncogenic signaling pathways. Indeed, agents targeting HSP90 function have shown therapeutic efficacy in several cancer types. We have examined the expression of HSP90 in different adrenal tumors and evaluated the use of HSP90 inhibitors in vitro as possible therapy for ACC. Methods: Immunohistochemical expression of HSP90 isoforms was investigated in different adrenocortical tumors and associated with clinical features. Additionally, a panel of N-terminal (17-allylamino-17-demethoxygeldanamycin (17-AAG), luminespib, and ganetespib) and C-terminal (novobiocin and silibinin) HSP90 inhibitors were tested on various ACC cell lines. Results: Within adrenocortical tumors, ACC samples exhibited the highest expression of HSP90β. Within a cohort of ACC patients, HSP90β expression levels were inversely correlated with recurrence-free and overall survival. In functional assays, among five different compounds tested luminespib and ganetespib induced a significant decrease in cell viability in single as well as in combined treatments with compounds of the clinically used EDP-M scheme (etoposide, doxorubicin, cisplatin, mitotane). Inhibition of cell viability correlated furthermore with a decrease in proliferation, in cell migration and an increase in apoptosis. Moreover, analysis of cancer pathways indicated a modulation of the ERK1/2—and AKT—pathways by luminespib and ganetespib treatment. Conclusions: Our findings emphasize HSP90 as a marker with prognostic impact and promising target with N-terminal HSP90 inhibitors as drugs with potential therapeutic efficacy toward ACC. KW - adrenal gland KW - cortisol KW - N-terminal HSP90 inhibitors KW - C-terminal HSP90 inhibitors KW - prognostic marker Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238029 VL - 10 ER - TY - JOUR A1 - Nguyen, Minh-Thu A1 - Saising, Jongkon A1 - Tribelli, Paula Maria A1 - Nega, Mulugeta A1 - Diene, Seydina M. A1 - François, Patrice A1 - Schrenzel, Jacques A1 - Spröer, Cathrin A1 - Bunk, Boyke A1 - Ebner, Patrick A1 - Hertlein, Tobias A1 - Kumari, Nimerta A1 - Härtner, Thomas A1 - Wistuba, Dorothee A1 - Voravuthikunchai, Supayang P. A1 - Mäder, Ulrike A1 - Ohlsen, Knut A1 - Götz, Friedrich T1 - Inactivation of farR Causes High Rhodomyrtone Resistance and Increased Pathogenicity in Staphylococcus aureus JF - Frontiers in Microbiology N2 - Rhodomyrtone (Rom) is an acylphloroglucinol antibiotic originally isolated from leaves of Rhodomyrtus tomentosa. Rom targets the bacterial membrane and is active against a wide range of Gram-positive bacteria but the exact mode of action remains obscure. Here we isolated and characterized a spontaneous Rom-resistant mutant from the model strain Staphylococcus aureus HG001 (RomR) to learn more about the resistance mechanism. We showed that Rom-resistance is based on a single point mutation in the coding region of farR [regulator of fatty acid (FA) resistance] that causes an amino acid change from Cys to Arg at position 116 in FarR, that affects FarR activity. Comparative transcriptome analysis revealed that mutated farR affects transcription of many genes in distinct pathways. FarR represses for example the expression of its own gene (farR), its flanking gene farE (effector of FA resistance), and other global regulators such as agr and sarA. All these genes were consequently upregulated in the RomR clone. Particularly the upregulation of agr and sarA leads to increased expression of virulence genes rendering the RomR clone more cytotoxic and more pathogenic in a mouse infection model. The Rom-resistance is largely due to the de-repression of farE. FarE is described as an efflux pump for linoleic and arachidonic acids. We observed an increased release of lipids in the RomR clone compared to its parental strain HG001. If farE is deleted in the RomR clone, or, if native farR is expressed in the RomR strain, the corresponding strains become hypersensitive to Rom. Overall, we show here that the high Rom-resistance is mediated by overexpression of farE in the RomR clone, that FarR is an important regulator, and that the point mutation in farR (RomR clone) makes the clone hyper-virulent. KW - antibiotic KW - Gram-positive bacteria KW - rhodomyrtone KW - Staphylococcus KW - membrane active Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224117 VL - 10 ER - TY - JOUR A1 - John, Cathy N. A1 - Abrantes, Pedro M. D. S. A1 - Prusty, Bhupesh K. A1 - Ablashi, Dharam V. A1 - Africa, Charlene W. J. T1 - K21 Compound, a Potent Antifungal Agent: Implications for the Treatment of Fluconazole-Resistant HIV-Associated Candida Species JF - Frontiers in Microbiology N2 - Background/Objectives: With mucocutaneous candidiasis being highly prevalent in HIV patients, the emergence of fluconazole-resistant Candida species forms a major challenge in treating and eradicating these infections. The objective of this study was to establish the antifungal activity of K21, a membrane-rupturing antimicrobial compound derived from a silica quaternary ammonium compound (SiQAC) with tetraethoxysilane (TEOS). Methods: The study sample included 81 Candida species of which 9 were type strains and 72 were clinical isolates. Minimum inhibitory concentrations, synergy, fractional inhibitory concentration index (FICI), and time kill assays were determined by broth microdilution. Electron microscopy (EM) was used to determine the qualitative changes brought about after treatment with K21. Results: K21 inhibited the growth of all fluconazole-resistant and susceptible Candida strains with only 2 h of exposure required to effectively kill 99.9% of the inoculum, and a definite synergistic effect was observed with a combination of K21 and fluconazole. EM demonstrated the presence of two forms of extracellular vesicles indicative of biofilm formation and cell lysis. Conclusion: The study established the efficacy of K21 as an antifungal agent and with fluconazole-resistant candidiasis on the increase, the development of K21 can provide a promising alternative to combat acquired drug resistance. KW - HIV-associated candidiasis KW - antimicrobial compounds KW - Candida KW - fluconazole KW - antifungal susceptibility KW - broth microdilution Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323505 VL - 10 ER - TY - THES A1 - Hofmann, Kristina Simone T1 - Untersuchung von mechanischen Eigenschaften, Bruchfestigkeit und Haftfestigkeit von Multilayer Zirkoniumdioxidkeramik zu dualhärtenden Befestigungskompositen T1 - Investigation of mechanical properties, fracture strength and shear bond strength of multilayer zirconia ceramics to dual-curing luting composites N2 - Ziel der Arbeit war es, den Einfluss der Platzierung der Restauration im Rohling auf die mechanischen Eigenschaften und die Scherfestigkeit zu dualhärtenden Befestigungskompositen zu untersuchen sowie Unterschiede in der Bruchfestigkeit zwischen zwei Multilayerkeramiken zu ermitteln. Material und Methodik: Es wurden 160 zylindrische Prüfkörper aus der Multilayerkeramik Katana Zirconia ML hergestellt, um mechanische Eigenschaften wie Dichte, Biegefestigkeit und Härte zu bestimmen. Eine Gruppe wurde künstlich gealtert. Die Bruchfestigkeit von 32 Kronen (Katana Zirconia ML, Ceramill Zolid FX Multilayer) wurde vor und nach thermischer sowie mechanischer Belastung untersucht. Zur Bestimmung der Scherfestigkeit wurden 512 quadratische Prüfkörper hergestellt, die verschiedenen thermischen Belastungen ausgesetzt wurden. Die Scherfestigkeit wurde mit einer Universalprüfmaschine bestimmt und die Brucharten (adhäsiv, kohäsiv, gemischt) wurden analysiert. Ergebnisse: Es gab keinen signifikanten Unterschied zwischen den Schichten oder der Alterung bei Dichte, Biegefestigkeit und Härte. Katana Zirconia ML zeigte höhere Bruchfestigkeit als Ceramill Zolid FX Multilayer. Die Scherfestigkeit von Panavia V5 zu Katana Zirconia ML war nicht durch thermische Belastung beeinflusst, jedoch bei Ceramill Zolid FX Multilayer. Der Haftverbund von Visalys CemCore war durch thermische Belastung beeinflusst, während Panavia V5 zu beiden Keramiken höhere Werte aufwies. Katana Zirconia ML hatte höhere Scherfestigkeitswerte als Ceramill Zolid FX Multilayer. Schlussfolgerung: Multilayerkeramik stellt eine arbeitsverringernde Alternative für den Seitenzahnbereich dar, ohne mechanische und ästhetische Einbußen. Die Platzierung im Rohling hat keinen Einfluss auf die Eigenschaften, jedoch ist der Haftverbund vom Befestigungskomposit abhängig. N2 - The aim of the work was to investigate the influence of the placement of the restoration in the blank on the mechanical properties and the shear bond strength of dual-curing luting composites and to determine differences in the fracture strength between two multilayer ceramics. Material and method: 160 cylindrical test specimens were made from the multilayer ceramic Katana Zirconia ML to determine mechanical properties such as density, flexural strength and hardness. One group was artificially aged. The breaking strength of 32 crowns (Katana Zirconia ML, Ceramill Zolid FX Multilayer) was examined before and after thermal and mechanical loading. To determine the shear bond strength, 512 square test specimens were produced and subjected to various thermal loads. The shear bond strength was determined using a universal testing machine and the fracture types (adhesive, cohesive, mixed) were analyzed. Results: There was no significant difference between layers or aging in density, flexural strength and hardness. Katana Zirconia ML showed higher breaking strength than Ceramill Zolid FX Multilayer. The shear bond strength of Panavia V5 to Katana Zirconia ML was not affected by thermal loading, but was affected in combination with Ceramill Zolid FX Multilayer. The adhesive bond of Visalys CemCore was influenced by thermal stress, while Panavia V5 had higher shear bond strength to both ceramics. Katana Zirconia ML had higher shear bond strength than Ceramill Zolid FX Multilayer. Conclusion: Multilayer ceramics represent a labor-reducing alternative for the posterior region, without mechanical and aesthetic losses. The placement in the blank has no influence on the properties, but the adhesive bond depends on the fixing composite. KW - Zirkoniumoxidkeramik KW - Multilayer-Zirkoniumdioxidkeramiken KW - Biaxiale Biegefestigkeit KW - Vickers-Härte KW - CAD/CAM restoration material KW - adhesive luting agent KW - shear bond strength KW - Biegefestigkeit KW - Härte KW - Mechanische Eigenschaft KW - Restaurative Zahnmedizin KW - Scherfestigkeit KW - Befestigungszement Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370308 ER - TY - JOUR A1 - Gronwald, Thomas A1 - Hoos, Olaf A1 - Hottenrott, Kuno T1 - Effects of Acute Normobaric Hypoxia on Non-linear Dynamics of Cardiac Autonomic Activity During Constant Workload Cycling Exercise JF - Frontiers in Physiology N2 - Aim: Measurements of Non-linear dynamics of heart rate variability (HRV) provide new possibilities to monitor cardiac autonomic activity during exercise under different environmental conditions. Using detrended fluctuation analysis (DFA) technique to assess correlation properties of heart rate (HR) dynamics, the present study examines the influence of normobaric hypoxic conditions (HC) in comparison to normoxic conditions (NC) during a constant workload exercise. Materials and Methods: Nine well trained cyclists performed a continuous workload exercise on a cycle ergometer with an intensity corresponding to the individual anaerobic threshold until voluntary exhaustion under both NC and HC (15% O2). The individual exercise duration was normalized to 10% sections (10–100%). During exercise HR and RR-intervals were continuously-recorded. Besides HRV time-domain measurements (meanRR, SDNN), fractal correlation properties using short-term scaling exponent alpha1 of DFA were calculated. Additionally, blood lactate (La), oxygen saturation of the blood (SpO2), and rating of perceived exertion (RPE) were recorded in regular time intervals. Results: We observed significant changes under NC and HC for all parameters from the beginning to the end of the exercise (10% vs. 100%) except for SpO2 and SDNN during NC: increases for HR, La, and RPE in both conditions; decreases for SpO2 and SDNN during HC, meanRR and DFA-alpha1 during both conditions. Under HC HR (40–70%), La (10–90%), and RPE (50–90%) were significantly-higher, SpO2 (10–100%), meanRR (40–70%), and DFA-alpha1 (20–60%) were significantly-lower than under NC. Conclusion: Under both conditions, prolonged exercise until voluntary exhaustion provokes a lower total variability combined with a reduction in the amplitude and correlation properties of RR fluctuations which may be attributed to increased organismic demands. Additionally, HC provoked higher demands and loss of correlation properties at an earlier stage during the exercise regime, implying an accelerated alteration of cardiac autonomic regulation. KW - autonomic nervous system KW - heart rate variability KW - detrended fluctuation analysis KW - endurance exercise KW - voluntary exhaustion KW - hypoxia Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369199 VL - 10 ER - TY - JOUR A1 - Fahmy-Garcia, Shorouk A1 - Farrell, Eric A1 - Witte-Bouma, Janneke A1 - Robbesom-van den Berge, Iris A1 - Suarez, Melva A1 - Mumcuoglu, Didem A1 - Walles, Heike A1 - Kluijtmans, Sebastiaan G. J. M. A1 - van der Eerden, Bram C. J. A1 - van Osch, Gerjo J. V. M. A1 - van Leeuwen, Johannes P. T. M. A1 - van Driel, Marjolein T1 - Follistatin Effects in Migration, Vascularization, and Osteogenesis in vitro and Bone Repair in vivo JF - Frontiers in Bioengineering and Biotechnology N2 - The use of biomaterials and signaling molecules to induce bone formation is a promising approach in the field of bone tissue engineering. Follistatin (FST) is a glycoprotein able to bind irreversibly to activin A, a protein that has been reported to inhibit bone formation. We investigated the effect of FST in critical processes for bone repair, such as cell recruitment, osteogenesis and vascularization, and ultimately its use for bone tissue engineering. In vitro, FST promoted mesenchymal stem cell (MSC) and endothelial cell (EC) migration as well as essential steps in the formation and expansion of the vasculature such as EC tube-formation and sprouting. FST did not enhance osteogenic differentiation of MSCs, but increased committed osteoblast mineralization. In vivo, FST was loaded in an in situ gelling formulation made by alginate and recombinant collagen-based peptide microspheres and implanted in a rat calvarial defect model. Two FST variants (FST288 and FST315) with major differences in their affinity to cell-surface proteoglycans, which may influence their effect upon in vivo bone repair, were tested. In vitro, most of the loaded FST315 was released over 4 weeks, contrary to FST288, which was mostly retained in the biomaterial. However, none of the FST variants improved in vivo bone healing compared to control. These results demonstrate that FST enhances crucial processes needed for bone repair. Further studies need to investigate the optimal FST carrier for bone regeneration. KW - follistatin 315 (FST315) KW - follistatin 288 (FST288) KW - migration KW - vascularization KW - osteogenesis KW - injectable in situ gelling slow release system KW - bone tissue engineering KW - regenerative medicine Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227617 VL - 7 ER - TY - JOUR A1 - Gál, Bernadett I. A1 - Kilencz, Tünde A1 - Albert, Anita A1 - Demeter, Ildikó A1 - Hegedűs, Klára Mária A1 - Janka, Zoltán A1 - Csifcsák, Gábor A1 - Álmos, Péter Z. T1 - Mild Effect of Nalmefene on Alcoholic Cue-Induced Response Invigoration in Alcohol Use Disorder Without Accompanying Changes in Electrophysiological Signatures of Early Visual Processing and Executive Control JF - Frontiers in Pharmacology N2 - Nalmefene is approved for as-needed pharmacological treatment in alcohol use disorder (AUD) by the European Medicines Agency. While the cellular effects of nalmefene have been thoroughly investigated, data are very limited on how this agent influences neural signals associated with inhibitory control and the visual analysis of environmental cues. This double-blind crossover study assessed the behavioral and neural effects of acute nalmefene administration in patients diagnosed with AUD. In experiment 1, we validated our experimental paradigm (electroencephalography combined with a modified Go/NoGo task using images of alcoholic and nonalcoholic drinks as prime stimuli) in 20 healthy adults to ensure that our protocol is suitable for assessing the behavioral and neural aspects of executive control. In experiment 2, we recruited 19 patients with AUD, and in a double-blind crossover design, we investigated the effects of nalmefene versus placebo on task performance (response accuracy, the sensitivity index, and reaction times), visual responses to appetitive cues (occipital P1, N1, and P2 components), and electrophysiological markers of conflict detection and response inhibition (frontal N2 and P3 waveforms). Under placebo, patients produced faster reaction times to alcohol-primed Go stimuli, an effect that was weak despite being statistically significant. However, the effect of alcoholic cues on the speed of response initiation disappeared after receiving nalmefene. We found no placebo versus nalmefene difference regarding our patients’ ability to accurately inhibit responses to NoGo stimuli or for occipital and frontal event-related potentials. Our results suggest that nalmefene might be potent in reducing the vigor to act upon alcoholic cues in AUD patients, but this effect is most probably mediated via subcortical (rather than cortical) neural circuits. KW - nalmefene KW - alcohol use disorder KW - response inhibition KW - incentive salience KW - event-related potentials KW - Go/NoGo task Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-369182 VL - 10 ER - TY - JOUR A1 - Franke, Maximilian A1 - Conzelmann, Annette A1 - Grünblatt, Edna A1 - Werling, Anna M. A1 - Spieles, Helen A1 - Wewetzer, Christoph A1 - Warnke, Andreas A1 - Romanos, Marcel A1 - Walitza, Susanne A1 - Renner, Tobias J. T1 - No Association of Variants of the NPY-System With Obsessive-Compulsive Disorder in Children and Adolescents JF - Frontiers in Molecular Neuroscience N2 - Obsessive-compulsive disorder (OCD) causes severe distress and is therefore counted by the World Health Organisation (WHO) as one of the 10 most impairing illnesses. There is evidence for a strong genetic underpinning especially in early onset OCD (eoOCD). Though several genes involved in neurotransmission have been reported as candidates, there is still a need to identify new pathways. In this study, we focussed on genetic variants of the Neuropeptide Y (NPY) system. NPY is one of the most abundant neuropeptides in the human brain with emerging evidence of capacity to modulate stress response, which is of high relevance in OCD. We focussed on tag-SNPs of NPY and its receptor gene NPY1R in a family-based approach. The sample comprised 86 patients (children and adolescents) with eoOCD with both their biological parents. However, this first study on genetic variants of the NPY-system could not confirm the association between the investigated SNPs and eoOCD. Based on the small sample size results have to be interpreted as preliminary and should be replicated in larger samples. However, also in an additional GWAS analysis in a large sample, we could not observe an associations between NPY and OCD. Overall, these preliminary results point to a minor role of NPY on the stress response of OCD. KW - NPY KW - obsessive-compulsive KW - children KW - anxiety KW - neuropeptide Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229051 VL - 12 ER - TY - JOUR A1 - Geran, Rohat A1 - Uecker, Florian C. A1 - Prüss, Harald A1 - Haeusler, Karl Georg A1 - Paul, Friedemann A1 - Ruprecht, Klemens A1 - Harms, Lutz A1 - Schmidt, Felix A. T1 - Olfactory and Gustatory Dysfunction in Patients With Autoimmune Encephalitis JF - Frontiers in Neurology N2 - Objective: To test the hypothesis that olfactory (OF) and gustatory function (GF) is disturbed in patients with autoimmune encephalitides (AE). Methods: The orthonasal OF was tested in 32 patients with AE and 32 age- and sex-matched healthy controls (HC) with the standardized Threshold Discrimination Identification (TDI) score. This validated olfactory testing method yields individual scores for olfactory threshold (T), odor discrimination (D), and identification (I), along with a composite TDI score. The GF was determined by the Taste Strip Test (TST). Results: Overall, 24/32 (75%) of patients with AE, but none of 32 HC (p < 0.001) had olfactory dysfunction in TDI testing. The results of the threshold, discrimination and identification subtests were significantly reduced in patients with AE compared to HC (all p < 0.001). Assessed by TST, 5/19 (26.3%) of patients with AE, but none of 19 HC presented a significant limitation in GF (p < 0.001). The TDI score was correlated with the subjective estimation of the olfactory capacity on a visual analog scale (VAS; rs = 0.475, p = 0.008). Neither age, sex, modified Rankin Scale nor disease duration were associated with the composite TDI score. Conclusions: This is the first study investigating OF and GF in AE patients. According to unblinded assessment, patients with AE have a reduced olfactory and gustatory capacity compared to HC, suggesting that olfactory and gustatory dysfunction are hitherto unrecognized symptoms in AE. Further studies with larger number of AE patients would be of interest to verify our results. KW - autoimmune encephalitis KW - olfactory dysfunction KW - gustatory dysfunction KW - olfactory testing KW - threshold discrimination identification test Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232921 VL - 10 ER - TY - JOUR A1 - Jarick, Katja J. A1 - Mokhtari, Zeinab A1 - Scheller, Lukas A1 - Hartweg, Julia A1 - Thusek, Sina A1 - Le, Duc-Dung A1 - Ranecky, Maria A1 - Shaikh, Haroon A1 - Qureischi, Musga A1 - Heinze, Katrin G. A1 - Beilhack, Andreas T1 - Photoconversion of Alloreactive T Cells in Murine Peyer’s Patches During Acute Graft-Versus-Host Disease: Tracking the Homing Route of Highly Proliferative Cells In Vivo JF - Frontiers in Immunology N2 - The regulation of immune cell migration throughout the body is essential to warrant immunosurveillance and to maintain immune homeostasis. Marking and tracking of these cells has proven important to study mechanisms of immune cell trafficking and cell interaction in vivo. Photoconversion is a well-suited technique for intravital application because it enables contactless time- and location-specific marking of cells in the tissue without surgically manipulating the microenvironment of the cells in question. However, in dividing cells the converted fluorescent protein may decline quickly. Here, we provide a detailed description of the photoconversion technique and its applicability to tracking highly proliferating T cells from the priming site of T cell activation to peripheral target organs of effector function in a preclinical model. Dendra2+ T cells were photoconverted in the Peyer’s patches during the initiation phase of acute graft-versus-host disease (GvHD) and tracked through the mesenteric lymph nodes and the peripheral blood to the small intestine with flow cytometry and intravital two-photon microscopy. Photoconverted alloreactive T cells preserved the full proliferative capacity, homing, and migration of alloreactive T cells in the intestinal lamina propria. We conclusively proved that photoconversion of highly proliferative alloreactive T cells in the Peyer’s patches is an effective tool to study trafficking of alloreactive T cells under physiologic conditions and to GvHD target tissues. This technique can also be applied to the study of immune cell tracking under inflammatory and non-inflammatory conditions. KW - T cell migration KW - acute graft-versus-host disease KW - mouse models KW - photoconversion KW - Dendra2 KW - Peyer's patch KW - in vivo cell tracking KW - lymphocyte homing Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323309 VL - 9 ER - TY - JOUR A1 - Herster, Franziska A1 - Bittner, Zsofia A1 - Codrea, Marius Cosmin A1 - Archer, Nathan K. A1 - Heister, Martin A1 - Löffler, Markus W. A1 - Heumos, Simon A1 - Wegner, Joanna A1 - Businger, Ramona A1 - Schindler, Michael A1 - Stegner, David A1 - Schäkel, Knut A1 - Grabbe, Stephan A1 - Ghoreschi, Kamran A1 - Miller, Lloyd S. A1 - Weber, Alexander N. R. T1 - Platelets Aggregate With Neutrophils and Promote Skin Pathology in Psoriasis JF - Frontiers in Immunology N2 - Psoriasis is a frequent systemic inflammatory autoimmune disease characterized primarily by skin lesions with massive infiltration of leukocytes, but frequently also presents with cardiovascular comorbidities. Especially polymorphonuclear neutrophils (PMNs) abundantly infiltrate psoriatic skin but the cues that prompt PMNs to home to the skin are not well-defined. To identify PMN surface receptors that may explain PMN skin homing in psoriasis patients, we screened 332 surface antigens on primary human blood PMNs from healthy donors and psoriasis patients. We identified platelet surface antigens as a defining feature of psoriasis PMNs, due to a significantly increased aggregation of neutrophils and platelets in the blood of psoriasis patients. Similarly, in the imiquimod-induced experimental in vivo mouse model of psoriasis, disease induction promoted PMN-platelet aggregate formation. In psoriasis patients, disease incidence directly correlated with blood platelet counts and platelets were detected in direct contact with PMNs in psoriatic but not healthy skin. Importantly, depletion of circulating platelets in mice in vivo ameliorated disease severity significantly, indicating that both PMNs and platelets may be relevant for psoriasis pathology and disease severity. KW - psoriasis KW - neutrophil KW - platelet KW - platelet-neutrophil complexes (PNCs) KW - imiquimod Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-320175 VL - 10 ER - TY - JOUR A1 - Kasaragod, Vikram B. A1 - Schindelin, Hermann T1 - Structure–Function Relationships of Glycine and GABAA Receptors and Their Interplay With the Scaffolding Protein Gephyrin JF - Frontiers in Molecular Neuroscience N2 - Glycine and γ-aminobutyric acid (GABA) are the major determinants of inhibition in the central nervous system (CNS). These neurotransmitters target glycine and GABAA receptors, respectively, which both belong to the Cys-loop superfamily of pentameric ligand-gated ion channels (pLGICs). Interactions of the neurotransmitters with the cognate receptors result in receptor opening and a subsequent influx of chloride ions, which, in turn, leads to hyperpolarization of the membrane potential, thus counteracting excitatory stimuli. The majority of glycine receptors and a significant fraction of GABAA receptors (GABAARs) are recruited and anchored to the post-synaptic membrane by the central scaffolding protein gephyrin. This ∼93 kDa moonlighting protein is structurally organized into an N-terminal G-domain (GephG) connected to a C-terminal E-domain (GephE) via a long unstructured linker. Both inhibitory neurotransmitter receptors interact via a short peptide motif located in the large cytoplasmic loop located in between transmembrane helices 3 and 4 (TM3-TM4) of the receptors with a universal receptor-binding epitope residing in GephE. Gephyrin engages in nearly identical interactions with the receptors at the N-terminal end of the peptide motif, and receptor-specific interaction toward the C-terminal region of the peptide. In addition to its receptor-anchoring function, gephyrin also interacts with a rather large collection of macromolecules including different cytoskeletal elements, thus acting as central scaffold at inhibitory post-synaptic specializations. Dysfunctions in receptor-mediated or gephyrin-mediated neurotransmission have been identified in various severe neurodevelopmental disorders. Although biochemical, cellular and electrophysiological studies have helped to understand the physiological and pharmacological roles of the receptors, recent high resolution structures of the receptors have strengthened our understanding of the receptors and their gating mechanisms. Besides that, multiple crystal structures of GephE in complex with receptor-derived peptides have shed light into receptor clustering by gephyrin at inhibitory post-synapses. This review will highlight recent biochemical and structural insights into gephyrin and the GlyRs as well as GABAA receptors, which provide a deeper understanding of the molecular machinery mediating inhibitory neurotransmission. KW - glycine receptors KW - GABAA receptors KW - gephyrin KW - moonlighting protein KW - inhibitory post-synaptic specialization KW - cytoskeletal proteins Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325607 VL - 11 ER - TY - JOUR A1 - Kervarrec, Thibault A1 - Samimi, Mahtab A1 - Guyétant, Serge A1 - Sarma, Bhavishya A1 - Chéret, Jérémy A1 - Blanchard, Emmanuelle A1 - Berthon, Patricia A1 - Schrama, David A1 - Houben, Roland A1 - Touzé, Antoine T1 - Histogenesis of Merkel Cell Carcinoma: A Comprehensive Review JF - Frontiers in Oncology N2 - Merkel cell carcinoma (MCC) is a primary neuroendocrine carcinoma of the skin. This neoplasia features aggressive behavior, resulting in a 5-year overall survival rate of 40%. In 2008, Feng et al. identified Merkel cell polyomavirus (MCPyV) integration into the host genome as the main event leading to MCC oncogenesis. However, despite identification of this crucial viral oncogenic trigger, the nature of the cell in which MCC oncogenesis occurs is actually unknown. In fact, several hypotheses have been proposed. Despite the large similarity in phenotype features between MCC tumor cells and physiological Merkel cells (MCs), a specialized subpopulation of the epidermis acting as mechanoreceptor of the skin, several points argue against the hypothesis that MCC derives directly from MCs. Alternatively, MCPyV integration could occur in another cell type and induce acquisition of an MC-like phenotype. Accordingly, an epithelial as well as a fibroblastic or B-cell origin of MCC has been proposed mainly based on phenotype similarities shared by MCC and these potential ancestries. The aim of this present review is to provide a comprehensive review of the current knowledge of the histogenesis of MCC. KW - merkel cell polyomavirus (MCPyV) KW - epithelial KW - fibroblast KW - B cell KW - Merkel cell carcinoma (MCC) KW - histogenesis KW - origin Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325733 VL - 9 ER - TY - JOUR A1 - Nagy, Magdolna A1 - van Geffen, Johanna P. A1 - Stegner, David A1 - Adams, David J. A1 - Braun, Attila A1 - de Witt, Susanne M. A1 - Elvers, Margitta A1 - Geer, Mitchell J. A1 - Kuijpers, Marijke J. E. A1 - Kunzelmann, Karl A1 - Mori, Jun A1 - Oury, Cécile A1 - Pircher, Joachim A1 - Pleines, Irina A1 - Poole, Alastair W. A1 - Senis, Yotis A. A1 - Verdoold, Remco A1 - Weber, Christian A1 - Nieswandt, Bernhard A1 - Heemskerk, Johan W. M. A1 - Baaten, Constance C. F. M. J. T1 - Comparative Analysis of Microfluidics Thrombus Formation in Multiple Genetically Modified Mice: Link to Thrombosis and Hemostasis JF - Frontiers in Cardiovascular Medicine N2 - Genetically modified mice are indispensable for establishing the roles of platelets in arterial thrombosis and hemostasis. Microfluidics assays using anticoagulated whole blood are commonly used as integrative proxy tests for platelet function in mice. In the present study, we quantified the changes in collagen-dependent thrombus formation for 38 different strains of (genetically) modified mice, all measured with the same microfluidics chamber. The mice included were deficient in platelet receptors, protein kinases or phosphatases, small GTPases or other signaling or scaffold proteins. By standardized re-analysis of high-resolution microscopic images, detailed information was obtained on altered platelet adhesion, aggregation and/or activation. For a subset of 11 mouse strains, these platelet functions were further evaluated in rhodocytin- and laminin-dependent thrombus formation, thus allowing a comparison of glycoprotein VI (GPVI), C-type lectin-like receptor 2 (CLEC2) and integrin α6β1 pathways. High homogeneity was found between wild-type mice datasets concerning adhesion and aggregation parameters. Quantitative comparison for the 38 modified mouse strains resulted in a matrix visualizing the impact of the respective (genetic) deficiency on thrombus formation with detailed insight into the type and extent of altered thrombus signatures. Network analysis revealed strong clusters of genes involved in GPVI signaling and Ca2+ homeostasis. The majority of mice demonstrating an antithrombotic phenotype in vivo displayed with a larger or smaller reduction in multi-parameter analysis of collagen-dependent thrombus formation in vitro. Remarkably, in only approximately half of the mouse strains that displayed reduced arterial thrombosis in vivo, this was accompanied by impaired hemostasis. This was also reflected by comparing in vitro thrombus formation (by microfluidics) with alterations in in vivo bleeding time. In conclusion, the presently developed multi-parameter analysis of thrombus formation using microfluidics can be used to: (i) determine the severity of platelet abnormalities; (ii) distinguish between altered platelet adhesion, aggregation and activation; and (iii) elucidate both collagen and non-collagen dependent alterations of thrombus formation. This approach may thereby aid in the better understanding and better assessment of genetic variation that affect in vivo arterial thrombosis and hemostasis. KW - arterial thrombus formation KW - bleeding KW - collagen KW - glycoprotein VI KW - platelets KW - microfluidics Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232194 VL - 6 ER - TY - THES A1 - Zhang, Tengyu T1 - Development of Modified polylysine based antibody conjugated nanoparticles with tumor-restricted, FcγR-independent stimulatory activity by targeting Fn14 T1 - Entwicklung modifizierter, mit Antikörpern konjugierter Nanopartikel auf Polylysinbasis mit tumorbeschränkter, FcγR- unabhängiger stimulierender Aktivität durch Ausrichtung auf Fn14 N2 - In this study, we developed an innovative nanoparticle formulation to facilitate the delivery of antitumor antibodies to tumor sites. The study commenced with the utilization of 13 bispecific antibody fusion proteins, which targeted the Fn14 receptor, thereby validating the pivotal role of crosslinking in Fn14 receptor activation. Subsequently, gold nanoparticles were activated using COOH-PEG-SH in combination with EDC/NHS, and subsequently conjugated with two Fn14-targeting antibodies, PDL192 and 5B6. Following this, a pH-sensitive shell was generated on the outer layer of the antibody-coupled gold nanoparticles through the application of chemically modified polylysine. The resultant complexes, termed MPL-antibody-AuNP, demonstrated a release profile reminiscent of the tumor microenvironment (TME). Notably, these complexes released antibody-AuNPs only in slightly acidic conditions while remaining intact in neutral or basic environments. Functionality analysis further affirmed the pH-sensitive property of MPL-antibody-AuNPs, demonstrating that the antibodies only initiated potent Fn14 activation in slightly acidic environments. This formulation holds potential for applicability to antibodies or ligands targeting the 80 TNFRSF family, given that gold nanoparticles successfully served as platforms for antibody crosslinking, thereby transforming these antibodies into potent agonists. Moreover, the TME disintegration profile of MPL mitigates the potential cytotoxic effects of antibodies, thereby circumventing associated adverse side effects. This study not only showcases the potential of nanoparticle formulations in targeted therapy, but also provides a solid foundation for further investigations on their clinical application in the context of targeting category II TNFRSF receptors with antibodies or ligands. N2 - In dieser Studie haben wir eine innovative Nanopartikel-Formulierung entwickelt, um die Auslieferung von Antitumor-Antikörpern an Tumorstellen zu erleichtern. Die Studie begann mit der Verwendung von 13 bispezifischen Antikörper-Fusionsproteinen, die auf den Fn14-Rezeptor abzielten, wodurch die entscheidende Rolle der Quervernetzung bei der Aktivierung des Fn14-Rezeptors bestätigt wurde. Anschließend wurden Goldnanopartikel unter Verwendung von COOH-PEG-SH in Kombination mit EDC/NHS aktiviert und danach mit zwei auf Fn14 abzielenden Antikörpern, PDL192 und 5B6, konjugiert. Daraufhin wurde eine pH-sensitive Schale auf der äußeren Schicht der mit Antikörpern gekoppelten Goldnanopartikel durch den Einsatz von chemisch modifiziertem Polylysin erzeugt. Die resultierenden Komplexe, bezeichnet als MPL-Antikörper-AuNP, zeigten ein Freisetzungsprofil, das an das Tumormikroumfeld (TME) erinnert. Bemerkenswert ist, dass diese Komplexe Antikörper-AuNP nur in leicht sauren Bedingungen freisetzten, während sie in neutralen oder basischen Umgebungen intakt blieben. Die Funktionalitätsanalyse bestätigte weiterhin die pH-empfindliche Eigenschaft der MPL-Antikörper-AuNPs, was zeigt, dass die Antikörper eine potente Fn14-Aktivierung nur in leicht sauren Bedingungen initiierten. Diese Formulierung hat Potenzial für die Anwendbarkeit auf Antikörper oder Liganden, die auf die Familie der TNFRSF abzielen, da die Goldnanopartikel erfolgreich als Plattformen für die Antikörpervernetzung dienten und diese Antikörper in potente Agonisten verwandelten. Darüber hinaus mildert das TME-Zerfallsprofil von MPL die potenziellen zytotoxischen Effekte der Antikörper, wodurch die damit verbundenen negativen Nebenwirkungen umgangen werden. Diese Studie zeigt nicht nur das Potenzial von Nanopartikel-Formulierungen in der gezielten Therapie auf, sondern bietet auch eine solide Grundlage für weitere Untersuchungen zu ihrer klinischen Anwendung im Kontext der Zielrichtung auf Kategorie-II-TNFRSF-Rezeptoren mit Antikörpern oder Liganden. KW - Immuntherapie KW - Immunotherapy Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358650 ER - TY - THES A1 - Hadi, Naji Said Aboud T1 - In vitro Studies on the Genotoxicity of Selected Pyrrolizidine Alkaloids T1 - In-vitro-Studien zur Genotoxizität ausgewählter Pyrrolizidinalkaloide N2 - Cancer is one of the leading causes of death worldwide. Toxic contaminants in human food or medicinal products, such as substances like pyrrolizidine alkaloids (PAs), have been thought to contribute to cancer incidence. PAs are found in many plant species as secondary metabolites, and they may affect humans through contaminated food sources, herbal medicines, and dietary supplements. Hundreds of compounds belonging to PAs have been identified, differing in their chemical structures, either in their necine base moiety or esterification at their necic acid moiety. PAs undergo hepatic metabolism, and after this process, they can induce hepatotoxicity, genotoxicity, and carcinogenicity. However, the mechanism of inducing genotoxicity and carcinogenicity is still unclear and warrants further investigation. Therefore, the present study aims to investigate the mechanism of genotoxicity induced by selected PAs with different chemical structures in in vitro systems. Primarily, human hepatoma HepG2 cells were utilized, and in co-culture, metabolically active HepG2 cells were combined with non-metabolically active human cervical HeLa H2B-GFP cells. First, the genotoxicity of the PAs europine, lycopsamine, retrorsine, riddelliine, seneciphylline, echimidine, and lasiocarpine was investigated in the cytokinesis-block micronucleus (CBMN) assay. All seven selected PAs caused the formation of micronuclei in a dose-dependent manner, with the maximal increase of micronucleus formation ranging from 1.64 to 2.0 fold. The lowest concentrations at which significant induction of micronuclei was found were 3.2 µM for lasiocarpine and riddelliine, 32 µM for retrorsine and echimidine, and 100 µM for seneciphylline, europine, and lycopsamine. These results confirmed previously published potency rankings in the micronucleus assay. The same PAs, with the exception of seneciphylline, were also investigated in a crosslink-modified comet assay, and reduced tail formation after hydrogen peroxide treatment was found in all diester-type PAs. Meanwhile, an equimolar concentration of the monoesters europine and lycopsamine did not significantly reduce DNA migration. Thus, the crosslinking activity was related to the ester type. Next, the role of metabolic enzymes and membrane transporters in PA-induced genotoxicity was assessed. Ketoconazole (CYP 450-3A4 inhibitor) prevented lasiocarpine-induced micronucleus formation completely, while furafylline (CYP 450-1A2 inhibitor) reduced lasiocarpine-induced micronucleus formation, but did not abolish it completely. This implies that the CYP 450 enzymes play an important role in PA-induced genotoxicity. Carboxylesterase 2 enzyme (CES 2) is commonly known to be involved in the detoxification of xenobiotics. Loperamide (CES 2 inhibitor) yielded an increased formation of lasiocarpine-induced micronuclei, revealing a possible role of CES-mediated detoxification in the genotoxicity of lasiocarpine. Also, intracellular glutathione (GSH) plays an important role in the detoxification of xenobiotics or toxins in the cells. Cells which had been pretreated with L-buthionine sulfoximine (BSO) to reduce GSH content were significantly more sensitive for the induction of micronucleus formation by lasiocarpine revealing the importance of GSH in PA-induced genotoxicity. Quinidine (Q) and nelfinavir (NFR) are OCT1 and OATP1B1 influx transporter inhibitors, respectively, which reduced micronucleus induction by lasiocarpine (only quinidine significantly), but not completely, pointing to a relevance of OCT1 for PA uptake in HepG2 cells. Verapamil (V) and benzbromarone (Bz) are MDR1 and MRP2 efflux transporter inhibitors, respectively, and they caused a slightly increased micronucleus induction by lasiocarpine (significant only for benzbromarone) thus, revealing the role of efflux transporters in PA-induced genotoxicity. The mechanistic approach to PA-induced genotoxicity was further studied based on oxidative stress via the formation of reactive oxygen species (ROS) in HepG2 cells. Overproduction of ROS can cross-link cellular macromolecules such as DNA, leading to genomic damage. An equimolar concentration of 10 µM of lasiocarpine (open-diester PA), riddelliine (cyclic-diester PA), and europine (monoester) significantly induced ROS production, with the highest ROS generation observed after lasiocarpine treatment, followed by riddelliine and then europine. No significant increase in ROS production was found with lycopsamine (10 µM; monoester PA), even at a higher concentration (320 µM). The generation of ROS by these PAs was further analyzed for confirmation by using 5 mM of the thiol radical scavenger antioxidant N-acetyl cysteine (NAC) combined with lasiocarpine, riddelliine, or europine. This analysis yielded a significant decrease in ROS after combining NAC with lasiocarpine, riddelliine, and europine. In addition, lasiocarpine, riddelliine, and europine induced a loss of mitochondrial membrane potential, pointing to mitochondria as the source of ROS generation. In vivo, hepatic sinusoidal epithelial cells (HSECs) are known to be damaged first by PAs after hepatic metabolization, but HSECs themselves do not express the required metabolic enzymes for activation of PAs. To mimic this situation, HepG2 cells were used to metabolically activate PA in a co-culture with HeLa H2B-GFP cells as non-metabolically active neighbours. Due to the green fluorescent GFP label the HeLa cells could be identified easily based in the co-culture. The PAs europine, riddelliine and lasiocarpine induced micronucleus formation in HepG2 cells, and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Metabolic inhibition of CYP 450 enzymes with ketoconazole abrogated micronucleus formation induced by the same PAs tested in the co-culture. The efflux transporter inhibitors verapamil and benzbromarone reduced the micronucleus formation in the co-culture. Furthermore, mitotic disturbances as an additional genotoxic mechanism of action were observed in HepG2 cells and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Overall, we were able to show that PAs were activated by HepG2 cells and the metabolites induced genomic damage in co-cultured non-metabolically active green HeLa cells. Finally, in HepG2 cells as well as the co-culture, combinations of PAs lasiocarpine and riddelliine favoured an additive effect rather than synergism. Thus, this study therefore provides support that the assumption of dose-addition can be applied in the characterization of the genotoxicity risk of PAs present in a mixture. N2 - Krebs ist eine der häufigsten Todesursachen weltweit. Toxische Verunreinigungen in Lebensmitteln oder pflanzlichen Arzneimitteln, wie Pyrrolizidinalkaloide (PAs), können zur Krebsinzidenz beitragen. PAs kommen in vielen Pflanzenarten als Sekundärmetabolite vor. Menschen können diese über kontaminierte Nahrungsquellen, pflanzliche Arzneimittel und Nahrungsergänzungsmittel aufnehmen. Eine Vielzahl von Verbindungen, die zu pyrrolizidinalkaloidhaltigen Substanzen (PAs) gehören, wurden identifiziert. Diese unterscheiden sich in ihrer chemischen Struktur entweder durch ihre Necinbaseneinheit oder ihre Veresterung an der Necicsäureeinheit. Nach metabolischer Aktivierung in der Leber können PAs Hepatotoxizität, Genotoxizität und Karzinogenität induzieren. Jedoch ist der Genotoxizitätsmechanismus nicht vollständig aufgeklärt und erfordert weitere Untersuchungen. Das Ziel dieser Studie liegt in der Untersuchung des Mechanismus der Genotoxizität, die in vitro durch bestimmte PAs mit unterschiedlicher chemischer Struktur induziert wird. Hierbei wurden primär humane Hepatom-HepG2-Zellen verwendet sowie in Co-Kultur metabolisch aktive HepG2-Zellen und nicht-metabolisch aktive humane zervikale HeLa H2B-GFP-Zellen. Zunächst wurde die Genotoxizität der PAs Europin, Lycopsamin, Retrorsin, Riddelliin, Seneciphyllin, Echimidin und Lasiocarpin im Zytokinese-Block-Mikronukleus-Assay (CBMN) untersucht. Die sieben (7) ausgewählten PAs führten dosisabhängig zur Bildung von Mikrokernen. Der maximale Anstieg der Mikronukleusbildung lag für alle PAs im Bereich des 1,64- bis 2,0-fachen des Ausgangswertes. Die niedrigsten Konzentrationen, bei denen eine signifikante Induktion von Mikrokernen gefunden wurde, waren 3,2 μM für Lasiocarpin und Riddelliin, 32 μM für Retrorsin und Echimidin sowie 100 μM für Seneciphyllin, Europin und Lycopsamin. Diese Ergebnisse bestätigen zuvor veröffentlichte Potenz-Rankings im Mikronukleus-Assay. Die Genotoxizität der gleichen PAs, mit Ausnahme von Seneciphyllin, wurde zusätzlich mittels eines Crosslink-modifizierten Comet-Assay untersucht. Es wurde eine reduzierte Schweifbildung nach der Behandlung mit Wasserstoffperoxid in allen PAs des Diestertyps gefunden, während eine äquimolare Konzentration der Monoester Europin und Lycopsamin die DNA-Migration nicht signifikant reduzierte. Dies deutet darauf hin, dass die Vernetzungsaktivität von PAs auf der Ester-Einheit beruht. Als nächstes wurde die Rolle von Stoffwechselenzymen und Membrantransportern in der PA-induzierten Genotoxizität untersucht. Ketoconazol (CYP 450-3A4-Inhibitor) verhinderte die Lasiocarpin-induzierte Mikronukleusbildung vollständig, während Furafyllin (CYP 450-1A2-Inhibitor) die Lasiocarpin-induzierte Mikronukleusbildung reduzierte, aber nicht vollständig beseitigte. Dies deutet darauf hin, dass CYP 450-Enzyme eine wichtige Rolle bei der PA-induzierten Genotoxizität spielen. Es ist allgemein bekannt, dass das Enzym Carboxylesterase 2 (CES-2) an der Entgiftung von Xenobiotika beteiligt ist. Loperamid (CES-2-Inhibitor) führte zu einer erhöhten Bildung von Lasiocarpin-induzierten Mikrokernen, was auf eine mögliche Rolle der CES-vermittelten Entgiftung bei der Genotoxizität von Lasiocarpin hindeutet. Auch intrazelluläres Glutathion (GSH) spielt eine wichtige Rolle bei der Entgiftung von Xenobiotika oder Toxinen. Zellen, die mit L-Buthioninsulfoximin (BSO) vorbehandelt worden waren, um den GSH-Gehalt zu reduzieren, waren signifikant empfindlicher für die Induktion der Mikronukleusbildung durch Lasiocarpin, was die Bedeutung von GSH für die PA-induzierte Genotoxizität zeigt. Chinidin (Q) und Nelfinavir (NFR) sind OCT1- bzw. OATP1B1-Influx-Transporter-Inhibitoren, die die Mikronukleus-Induktion durch Lasiocarpin reduzierten (nur Chinidin signifikant), aber nicht vollständig, was auf eine Relevanz von OCT1 für die PA-Aufnahme in HepG2-Zellen hindeutet.Verapamil (V) und Benzbromaron (Bz) sind MDR1- bzw. MRP2-Efflux-Transporter-Inhibitoren und verursachten eine leicht erhöhte Mikronukleus-Induktion durch Lasiocarpin (signifikant nur für Benzbromaron), was die Rolle von Efflux-Transportern bei der PA-induzierten Genotoxizität aufzeigt. Der Mechanismus der PA-induzierten Genotoxizität wurde auf der Grundlage von oxidativem Stress durch die Bildung von reaktiven Sauerstoffspezies (ROS) in HepG2-Zellen weiter untersucht. Eine Überproduktion von ROS kann zelluläre Makromoleküle wie DNA vernetzen, was zu genomischen Schäden führt. Eine äquimolare Konzentration von 10 μM von Lasiocarpin (Open-Diester PA), Riddelliin (Cyclic-Diester PA) und Europin (Monoester) induzierte signifikant die ROS-Produktion, wobei die höchste ROS-Erzeugung nach Lasiocarpin-Behandlung beobachtet wurde, gefolgt von Riddelliin und Europin. Mit Lycopsamin (10 μM; Monoester PA) wurde auch bei höherer Konzentration (320 μM) keine signifikante Steigerung der ROS-Produktion gefunden. Um die Beteiligung von ROS am Mechanismus der Genotoxizität einzelner PAs genauer zu betrachten und die bisherigen Ergebnisse zu bestätigen, wurden weitere Untersuchungen in Anwesenheit des Sauerstoffradikalfängers N-Acetylcysteine (NAC) in Kombination mit Lasiocarpin, Riddelliin oder Europin durchgeführt. Diese Analyse ergab eine signifikante Abnahme der ROS-Produktion nach der Kombination von NAC mit Lasiocarpin, Riddelliin und Europin. Darüber hinaus induzierten Lasiocarpin, Riddelliin und Europin Veränderungen im mitochondrialen Membranpotenzial. Dies deutet darauf hin, dass ROS vermehrt in den Mitochondrien der Zellen gebildet werden. Aus in vivo Daten ist bekannt, dass hepatische sinusoidale Epithelzellen (HSECs) die Zelltypen innerhalb der Leber sind, die nach der metabolischen Aktivierung von PAs zuerst geschädigt werden. Jedoch exprimieren HSECs nicht die erforderlichen Stoffwechselenzyme für die Aktivierung von PAs. Um diese Situation nachzuahmen, wurden HepG2-Zellen verwendet, um PAs in einer Kokultur mit HeLa H2B-GFP-Zellen als nicht-metabolisch aktive Nachbarn metabolisch zu aktivieren. Durch die grün fluoreszierende GFP-Markierung konnten die HeLa-Zellen in der Co-Kultur leicht identifiziert werden. Die PAs Europine, Riddelliin und Lasiocarpin induzierten die Bildung von Mikrokernen in HepG2-Zellen und in HeLa H2B-GFP-Zellen, die mit HepG2-Zellen kokultiviert wurden, jedoch nicht in HeLa H2B-GFP-Zellen, die allein kultiviert wurden. Die metabolische Hemmung von CYP 450-Enzymen mit Ketoconazol hob die Mikronukleusbildung, welche durch die zuvor getesteten PAs induziert wurde, auf. Die Efflux-Transporter-Inhibitoren Verapamil und Benzbromaron reduzierten die Mikronukleusbildung in der Kokultur. Darüber hinaus wurden mitotische Störungen als zusätzlicher genotoxischer Wirkmechanismus in der Co-Kultur aus HepG2-Zellen und in HeLa H2B-GFP-Zellen beobachtet, jedoch nicht in HeLa H2B-GFP-Zellen, die allein kultiviert wurden. Zusammengefasst deuten diese Ergebnisse darauf hin, dass PAs durch HepG2-Zellen bioaktiviert werden können und aus PAs gebildete Metabolite genomische Schäden in kokultivierten, nicht-metabolisch aktiven HeLa-Zellen induzierten. Abschließend zeigen Kombinationen der PAs Lasiocarpin und Riddelliin sowohl in HepG2-Zellen als auch in der Co-Kultur eher einen additiven Effekt als einen Synergismus. Diese Studie liefert daher Unterstützung für die Annahme, dass die Dosisaddition zur Charakterisierung des genotoxischen Risikos von in einem Gemisch vorhandenen PAs angewendet werden kann. KW - Pyrrolizidine alkaloids KW - HeLa H2B-GFP-Zellen KW - Pyrrolizidinalkaloide KW - Kleinkern KW - Mutagenität KW - Genotoxizität KW - DNA-Vernetzung KW - mitotische Störung KW - Co-culture KW - metabolische Aktivierung KW - Membrantransporter KW - metabolische Enzyme KW - HepG2-Zellen KW - Genotoxicity KW - Micronuclei KW - DNA crosslink KW - Mitotic disturbance KW - Metabolic activation KW - Membrane transporters KW - Metabolic enzymes KW - HepG2 cells KW - HeLa H2B-GFP cells KW - micronucleus Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-370376 ER - TY - THES A1 - Choi, Jihyoung T1 - Development of an Add-On Electrode for Non-Invasive Monitoring in Bioreactor Cultures and Medical Devices T1 - Entwicklung einer Zusatzelektrode für das nicht-invasive Monitoring von Bioreaktorkulturen und Medizinprodukten N2 - Electrochemical impedance spectroscopy (EIS) is a valuable technique analyzing electrochemical behavior of biological systems such as electrical characterization of cells and biomolecules, drug screening, and biomaterials in biomedical field. In EIS, an alternating current (AC) power signal is applied to the biological system, and the impedance of the system is measured over a range of frequencies. In vitro culture models of endothelial or epithelial barrier tissue can be achieved by culturing barrier tissue on scaffolds made with synthetic or biological materials that provide separate compartments (apical and basal sides), allowing for further studies on drug transport. EIS is a great candidate for non-invasive and real-time monitoring of the electrical properties that correlate with barrier integrity during the tissue modeling. Although commercially available transendothelial/transepithelial electrical resistance (TEER) measurement devices are widely used, their use is particularly common in static transwell culture. EIS is considered more suitable than TEER measurement devices in bioreactor cultures that involve dynamic fluid flow to obtain accurate and reliable measurements. Furthermore, while TEER measurement devices can only assess resistance at a single frequency, EIS measurements can capture both resistance and capacitance properties of cells, providing additional information about the cellular barrier's characteristics across various frequencies. Incorporating EIS into a bioreactor system requires the careful optimization of electrode integration within the bioreactor setup and measurement parameters to ensure accurate EIS measurements. Since bioreactors vary in size and design depending on the purpose of the study, most studies have reported using an electrode system specifically designed for a particular bioreactor. The aim of this work was to produce multi-applicable electrodes and established methods for automated non-invasive and real-time monitoring using the EIS technique in bioreactor cultures. Key to the electrode material, titanium nitride (TiN) coating was fabricated on different substrates (materials and shape) using physical vapor deposition (PVD) and housed in a polydimethylsiloxane (PDMS) structure to allow the electrodes to function as independent units. Various electrode designs were evaluated for double-layer capacitance and morphology using EIS and scanning electron microscopy (SEM), respectively. The TiN-coated tube electrode was identified as the optimal choice. Furthermore, EIS measurements were performed to examine the impact of influential parameters related to culture conditions on the TiN-coated electrode system. In order to demonstrate the versatility of the electrodes, these electrodes were then integrated into in different types of perfusion bioreactors for monitoring barrier cells. Blood-brain barrier (BBB) cells were cultured in the newly developed dynamic flow bioreactor, while human umblical vascular endothelial cells (HUVECs) and Caco-2 cells were cultured in the miniature hollow fiber bioreactor (HFBR). As a result, the TiN-coated tube electrode system enabled investigation of BBB barrier integrity in long-term bioreactor culture. While EIS measurement could not detect HUVECs electrical properties in miniature HFBR culture, there was the possibility of measuring the barrier integrity of Caco-2 cells, indicating potential usefulness for evaluating their barrier function. Following the bioreactor cultures, the application of the TiN-coated tube electrode was expanded to hemofiltration, based on the hypothesis that the EIS system may be used to monitor clotting or clogging phenomena in hemofiltration. The findings suggest that the EIS monitoring system can track changes in ion concentration of blood before and after hemofiltration in real-time, which may serve as an indicator of clogging of filter membranes. Overall, our research demonstrates the potential of TiN-coated tube electrodes for sensitive and versatile non-invasive monitoring in bioreactor cultures and medical devices. N2 - Die elektrochemische Impedanzspektroskopie (EIS) ist eine nützliche Methode, um das elektrochemische Verhalten von biologischen Systemen zu analysieren, wie z.B. die elektrische Charakterisierung von Zellen und Biomolekülen, Drug Screening und Biomaterialien im biomedizinischen Bereich. Für die EIS wird ein Wechselstrom an das biologische System angeschlossen und die Impedanz des Systems über einen Frequenzbereich gemessen. In vitro-Modelle von Gewebekulturen epithelialer Barrieren können mithilfe künstlicher oder biologischer Materialien, die über unterschiedliche Kompartimente (apikale und basolaterale Seite) verfügen, hergestellt werden und ermöglichen weitere Untersuchungen zum Transport von Arzneistoffen. Die EIS bietet dabei eine hervorragende Methode für das nicht-invasive Echtzeit-Monitoring der elektrischen Eigenschaften, die mit der Barriere-Integrität während der Gewebeentwicklung korreliert. Obwohl kommerziell erhältliche Geräte zur Messung des transendothelialen/transepithelialen elektrischen Widerstands (TEER) umfangreich verwendet werden, ist ihre Verwendung besonders bei statischen Transwell-Kulturen verbreitet. Durch die EIS kann im Gegensatz zur TEER-Messung für Bioreaktor-Kulturen, die einen dynamischen Medienfluss aufweisen, genauere und verlässliche Messungen erhalten werden. Zudem können EIS-Messungen anders als die TEER-Messung, die nur den Widerstand einer einzelnen Frequenz misst, gleichzeitig den elektrischen Widerstand und die Kapazität von Zellen erfassen und damit zusätzliche Informationen über die zellulären Barriereeigenschaften über verschiedene Frequenzen hinweg liefern. Der EIS-Einbau in ein Bioreaktor-System bedarf einer sorgfältigen Optimierung der Elektrodenintegration in das Bioreaktor-Setup und der Messparameter, um akkurate EIS-Messungen durchführen zu können. Da Bioreaktoren abhängig vom Untersuchungszweck in ihrer Größe und ihrem Design variieren, verwenden die meisten Studien speziell entwickelte Elektrodensysteme für einzelne Bioreaktoren. Das Ziel dieser Arbeit war die Herstellung von vielseitig anwendbaren Elektroden und etablierten Methoden für das automatisierte nicht-invasive Echtzeit-Monitoring von Bioreaktor-Kulturen mithilfe der EIS. Entscheidend für das Elektrodenmaterial war die Titannitrid (TiN)-Beschichtung, die auf verschiedenen Substraten (Materialien und Formen) durch Physical Vapor Deposition (PVD) hergestellt und in einer Polydimethylsiloxan (PDMS)-Struktur untergebracht wurde, damit die Elektroden unabhängig voneinander arbeiten können. Verschiedene Elektrodendesigns wurden auf Doppelschicht-Kapazität mithilfe der EIS bzw. auf die Morphologie mit Rasterelektronenmikroskopie untersucht. Die TiN-beschichteten Elektroden in Röhrenform erwiesen sich als optimal. Weiterhin wurden EIS-Messungen durchgeführt, um die Auswirkung von beeinflussenden Parametern auf die Kulturbedingungen durch das TiN-beschichtete Elektrodensystem zu untersuchen. Um die Vielseitigkeit der Elektroden aufzuzeigen, wurden diese anschließend zum Monitoring von Barriere-bildenden Zellen in unterschiedliche Perfusionsbioreaktoren integriert. Zellen der Blut-Hirn-Schranke (BHS) wurden im neu entwickelten dynamischen Flussreaktor kultiviert, wohingegen humane umbilikale vaskuläre Endothelzellen (HUVEC) und Caco-2-Zellen in Hohlfaserbioreaktoren (HFBR) in Miniaturform kultiviert wurden. Das TiN-beschichtete Röhrenelektrodensystem ermöglichte die Untersuchung der BHS-Barrieren-Integrität in einer Langzeit-Bioreaktorkultur. Während die EIS-Messung in der Miniaturform-HFBR-Kultur keine elektrischen Eigenschaften der HUVECs detektieren konnte, war es möglich, eine Barriere-Integrität der Caco-2-Zellen zu messen, die den potentiellen Nutzen für die Evaluierung deren Barrierefunktion aufzeigt. Nach den Bioreaktorkulturen wurde die Anwendung der TiN-beschichteten Röhrenelektrode auf die Hämofiltration erweitert, auf Grundlage der Hypothese, dass das EIS-System ein Gerinnen oder Verstopfen während der Hämofiltration überwachen könnte. Die Ergebnisse zeigen, dass das EIS-Monitoring-System Veränderungen in der Ionenkonzentration des Blutes vor und nach Hämofiltration in Echtzeit verfolgen kann, welches eventuell als Messgröße für ein Verstopfen der Filtermembranen genutzt werden kann. Insgesamt weisen TiN-beschichtete Röhrenelektroden unseren Forschungen zufolge ein großes Potential für ein empfindliches und vielfältiges nicht-invasives Monitoring von Bioreaktorkulturen und Medizingeräte auf. KW - Monitoring KW - Tissue Engineering KW - Electrode KW - Perfusion Bioreactor KW - Hemofiltration KW - Medizinprodukt KW - Electrochemical Impedance Spectroscopy Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358232 ER - TY - THES A1 - Kutschka, Ilona T1 - Activation of the integrated stress response induces remodeling of cardiac metabolism in Barth Syndrome T1 - Aktivierung der "Integrated Stress Response" führt zur Umstellung des kardialen Metabolismus im Barth Syndrom N2 - Barth Syndrome (BTHS) is an inherited X-chromosomal linked disorder, characterized by early development of cardiomyopathy, immune system defects, skeletal muscle myopathy and growth retardation. The disease displays a wide variety of symptoms including heart failure, exercise intolerance and fatigue due to the muscle weakness. The cause of the disease are mutations in the gene encoding for the mitochondrial transacylase Tafazzin (TAZ), which is important for remodeling of the phospholipid cardiolipin (CL). All mutations result in a pronounced decrease of the functional enzyme leading to an increase of monolysocardiolipin (MLCL), the precursor of mature CL, and a decrease in mature CL itself. CL is a hallmark phospholipid of mitochondrial membranes, highly enriched in the inner mitochondrial membrane (IMM). It is not only important for the formation of the cristae structures, but also for the function of different protein complexes associated with the mitochondrial membrane. Reduced levels of mature CL cause remodeling of the respiratory chain supercomplexes, impaired respiration, defects in the Krebs cycle and a loss of mitochondrial calcium uniporter (MCU) protein. The defective Ca2+ handling causes impaired redox homeostasis and energy metabolism resulting in cellular arrhythmias and defective electrical conduction. In an uncompensated situation, blunting mitochondrial Ca2+ uptake provokes increased mitochondrial emission of H2O2 during workload transitions, related to oxidation of NADPH, which is required to regenerate anti-oxidative enzymes. However, in the hearts and cardiac myocytes of mice with a global knock-down of the Taz gene (Taz-KD), no increase in mitochondrial ROS was observed, suggesting that other metabolic pathways may have compensated for reduced Krebs cycle activation. The healthy heart produces most of its energy by consuming fatty acids. In this study, the fatty acid uptake into mitochondria and their further degradation was investigated, which showed a switch of the metabolism in general in the Taz-KD mouse model. In vivo studies revealed an increase of glucose uptake into the heart and decreased fatty acid uptake and oxidation. Disturbed energy conversion resulted in activation of retrograde signaling pathways, implicating overall changes in the cell metabolism. Upregulated integrated stress response (ISR) was confirmed by increased levels of the downstream target, i.e., the activating transcription factor 4 (ATF4). A Tafazzin knockout mouse embryonal fibroblast cell model (TazKO) was used to inhibit the ISR using siRNA transfection or pharmaceutical inhibition. This verified the central role of II the ISR in regulating the metabolism in BTHS. Moreover, an increased metabolic flux into glutathione biosynthesis was observed, which supports redox homeostasis. In vivo PET-CT scans depicted elevated activity of the xCT system in the BTHS mouse heart, which transports essential amino acids for the biosynthesis of glutathione precursors. Furthermore, the stress induced signaling pathway also affected the glutamate metabolism, which fuels into the Krebs cycle via -ketoglutarate and therefore supports energy converting pathways. In summary, this thesis provides novel insights into the energy metabolism and redox homeostasis in Barth syndrome cardiomyopathy and its regulation by the integrated stress response, which plays a central role in the metabolic alterations. The aim of the thesis was to improve the understanding of these metabolic changes and to identify novel targets, which can provide new possibilities for therapeutic intervention in Barth syndrome. N2 - Barth Syndrome (BTHS) ist eine X-chromosomal vererbbare Erkrankung, welche sich in der frühen Entstehung von Kardiomyopathie, Störungen des Immunsystems, Skelettmuskelschwäche und Wachstumsverzögerungen manifestiert. Das Krankheitsbild ist sehr variabel mit milden Symptomen bis hin zu sehr schwerwiegenden Fällen, bei denen die schnelle Verschlechterung der Kardiomyopathie bereits in jungen Jahren eine Herztransplantation erfordern kann. Betroffenen Patienten zeigen eine deutliche Intoleranz gegenüber körperlicher Anstrengung, welche mit schneller Müdigkeit einhergeht. Die Krankheit wird durch verschiedene Mutationen auf dem Gen für die mitochondriale Transacylase Tafazzin (TAZ) ausgelöst. Die Mutationen führen zu einem Funktionsverlust des Enzyms, welches in der Biosynthese des Phospholipids Cardiolipin (CL) eine entscheidende Rolle spielt. Die Vorstufe des Lipids, das sogenannte Monolysocardiolipin (MLCL), reichert sich dadurch an, wohingegen die Menge an reifem CL entscheidend verringert ist. CL ist ein bedeutendes Phospholipid in den Mitochondrien, wo es vor allem in der inneren Mitochondrien Membran vorkommt. CL ist einerseits wichtig für die Ausbildung der Cristae Strukturen der inneren Mitochondrien Membran. Darüber hinaus ist es notwendig für die Struktur und Funktion verschiedenster Proteinkomplexe in der Membran, welche dadurch erst ihre volle Funktionsfähigkeit erhalten. Es wurde bereits gezeigt, dass der Verlust von reifem CL in BTHS zu einer Dissoziation der Superkomplexe der Atmungskette führt, welche dadurch in ihrer Funktion beeinträchtigt ist. Zusätzlich sind Störungen im Krebs Zyklus und der Kalziumaufnahme durch den mitochondriellen Kalzium (Ca2+) -Uniporter (MCU) Komplex bekannt. Die beeinträchtigte mitochondriale Ca2+ Aufnahme beeinflusst sowohl die Redox Homöostase als auch den Energie Metabolismus, was zu Arrhythmien und einer Störung der elektrischen Weiterleitung im Herzen führt. Im gesunden Herzen gewinnen die Herzmuskelzellen den Hauptanteil ihrer Energie aus dem Abbau von Fettsäuren. In dieser Studie wurde durch die Untersuchung des Fettsäurestoffwechsels im Taz knockdown Mausmodell (Taz-KD) gezeigt, dass eine deutliche Reduktion in Proteinen vorliegt, welche für die Aufnahme und die Verstoffwechselung der Fettsäuren in den Mitochondrien verantwortlich sind. Diese Veränderungen führten in vivo zu einer verringerten Aufnahme und Verstoffwechselung von Fettsäuren und einer Erhöhten Aufnahme von Glucose. Dysfunktionale Mitochondrien aktivieren retrograde Signalwege, welche eine generelle IV Umstellung des Metabolismus zur Folge haben. Eine erhöhte Menge des Transkriptionsfaktors ATF4, welcher sowohl Fettsäure- als auch Aminosäuremetabolismus beeinflusst, zeigte die Aktivierung der sogenannten „Integrated stress response“ (ISR). Ein Zellmodel embryonaler Fibroblasten aus der Maus mit einem Taz knockout (TazKO) wurde verwendet um die ISR durch siRNA Transfektion oder einem pharmakologischen Inhibitor zu blockieren. Dadurch konnte die zentrale Rolle der ISR in der Umstellung des Metabolismus bestätigt werden. Zusätzlich konnte eine erhöhte metabolische Aktivität in Richtung der Glutathion Biosynthese beobachtet werden, welche für die Redox Homöostase in den Mitochondrien von Bedeutung ist. In vivo PET-CT Untersuchungen zeigten eine erhöhte Aktivität des xCT Systems im Herzen des BTHS Mausmodells auf. Dies dient der Aufnahme von Aminosäuren, welche für die Glutathion Biosynthese benötigt werden. Hinzu kommt, dass die Aktivierung des Stresssignalweges den Glutamat Stoffwechsel in der Zelle beeinflusste. Über -Ketoglutarat trägt Glutamat so vermehrt zur Energiegewinnung bei. Das Ziel dieser Doktorarbeit war es, die metabolischen Veränderungen in BTHS zu untersuchen, um die veränderten Vorgänge besser zu verstehen und so neue mögliche Angriffspunkte für Therapiemöglichkeiten zu identifizieren. KW - Herzmuskelkrankheit KW - Mitochondrium KW - Stoffwechsel KW - Barth Syndrome Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358186 ER - TY - THES A1 - Wilhelmi, Kai Alexander T1 - Untersuchung von Veränderungen der myelinisierten Nervenfasern durch Entmarkung in Haut- und Nervenbiopsien von Patienten mit Polyneuropathie T1 - Examination of changes in myelinated nerve fibers due to demyelination in skin and nerve biopsies of patients with polyneuropathy N2 - In dieser Arbeit wurde durch das immunhistochemische Anfärben von nodalen (Natriumkanäle, NF), paranodalen (Caspr, NF) und internodalen (MBP) Proteinen der in Fingerhautbiopsien vorhanden Nervenfasern untersucht, ob eine Veränderung der typischen Verteilungsmuster dieser Proteine, eine demyelinisierende Polyneuropathie anzeigen kann. Dazu wurden am Universitätsklinikum Würzburg prospektiv 93 Polyneuropathie-Patienten und 25 Kontrollpersonen rekrutiert. Bei allen Patienten wurden Hautstanzbiospien am Zeigefinger durchgeführt. Bei 35 Patienten mit schweren oder unklaren Verläufen, wurden konsiliarisch Nervus suralis Biopsien durchgeführt. Aus einem Abschnitt von 27 dieser Biopsien, konnten im Rahmen dieser Arbeit Zupfnervenpräparate angefertigt und analog zu den Hautbiopsien ausgewertet werden. Aus der Routinediagnostik der Klinik flossen weiterhin die Ergebnisse der elektrophysiologischen Routinediagnostik und der Histologiebefund der Nervus suralis Biopsien in die Auswertung ein. Zusammenfassend kamen veränderte Natriumkanalbanden in Fingerhautbiopsien signifikant häufiger bei Patienten mit elektrophysiologisch als demyelinisierend befundeten Polyneuropathien, als bei Patienten mit elektrophysiologisch als axonal befundeten Polyneuropathien vor. Vielfach fanden sich veränderte Natriumkanalbanden inmitten para- und internodal unauffälliger Schnürringe und umgekehrt. Diese Beobachtung stützt die bereits in Vorarbeiten vorgeschlagene und in der aktuellen Leitlinie zur Diagnostik für Polyneuropathien aufgegriffene Entität der Paranodopathien (Uncini, Susuki, & Yuki, 2013). Möglich wäre, dass eine veränderte Verteilung der Natriumkanäle die schnelle Leitfähigkeit beeinträchtigen und somit trotz intakter Bemarkung, elektrophysiologisch das Bild einer demyelinisierenden Neuropathie vermittelt. Ein direkter Zusammenhang zwischen dem Auftreten von doppelten und verlängerten Natriumkanalbanden und einzelnen Messwerten (z.B. Amplituden und Latenzzeiten) fand sich nicht. Auch in den Zupfnervenpräparaten der Nervus suralis Biopsien, konnten o.g. Verteilungsmuster untersucht werden. Deren Vorkommen zeigte sich als unabhängig vom elektrophysiologischen und histologischen Befund, von der Ätiologie der PNP und von den gefundenen Veränderungen in den Hautbiopsien des betreffenden Patienten. N2 - Myelinated nerve fibers in finger skin biopsies and sural nerve biopsies were examined using immunohistochemical staining to detect changes in the typical distribution patterns of nodal (voltage-gated sodium channels, neurofascin 186), paranodal (Caspr, neurofascin 155), and internodal (myelin basic protein) proteins, aiming to identify indicators for demyelinating polyneuropathies. A total of 93 polyneuropathy patients and 25 control subjects were prospectively recruited from the University Hospital Würzburg. Skin punch biopsies were conducted on all patients and control subjects. Additionally, sural nerve biopsies were performed on a consultative basis for 35 patients. Teased nerve fiber preparations were made from a section of 27 of these biopsies and evaluated similarly to the skin biopsies. In summary, altered sodium channel bands in myelinated nerve fibers from finger skin biopsies were significantly more prevalent in patients electrophysiologically diagnosed with demyelinating polyneuropathies. However, there was no significant difference in the means of individual electrophysiological measurements between patients with and without changes in the immunohistochemical stainings. Each of the investigated changes was significantly more common in the polyneuropathy group than in the control group. Further correlations, particularly in the comparison of results from skin and sural nerve biopsies, were not found KW - Polyneuropathie KW - Ranvier-Schnürring KW - Entmarkung KW - Elektrophysiologie KW - Caspr KW - Neurofascin KW - MBP KW - spannungsgesteuerter Natriumkanal Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360046 ER - TY - JOUR A1 - Gerlich, C. A1 - Andreica, I. A1 - Küffner, R. A1 - Krause, D. A1 - Lakomek, H. J. A1 - Reusch, A. A1 - Braun, J. T1 - Evaluation einer Basisschulung für Patienten mit rheumatoider Arthritis T1 - Evaluation of a basic educational program for patients with rheumatoid arthritis JF - Zeitschrift für Rheumatologie N2 - Hintergrund Ein neues Rahmenkonzept hat die flexible Ableitung und Nutzung von rheumatologischen Schulungsprogrammen für unterschiedliche Versorgungsbereiche ermöglicht. Auf dieser Grundlage wurde eine 5‑stündige Basisschulung für Patienten mit rheumatoider Arthritis (RA) entwickelt, es wurden rheumatologische Fachärzte und Psychologen trainiert, und dann wurde die Wirksamkeit nach dem Wirkmodell der Patientenschulung evaluiert. Methoden Mit dem Studiendesign einer extern randomisierten Wartekontrollgruppenstudie mit 3 Messzeitpunkten wurde geprüft, wie sich die 5‑stündige Basisschulung auf das Erkrankungs- und Behandlungswissen sowie auf die Gesundheitskompetenz von RA-Patienten (n = 249) auswirkt. Weitere Fragen betrafen Einstellungsparameter, Kommunikationskompetenz, Erkrankungsauswirkungen und die Zufriedenheit mit der Schulung. Die Auswertungen erfolgten auf Intention-to-treat-Basis mit Kovarianzanalysen für die Hauptzielgrößen unter Berücksichtigung des Ausgangswertes. Ergebnisse Die Analysen zeigen, dass die Basisschulung RA wirksam ist. Noch 3 Monate nach der Schulung verfügten die Schulungsteilnehmer über mehr Wissen und Gesundheitskompetenz als die Wartekontrollgruppe mit kleinem bis mittelgroßem Effekt (d = 0,37 bzw. 0,38). In den Nebenzielgrößen zeigten sich mit Ausnahme der Krankheitskommunikation keine weiteren Schulungseffekte. Diskussion Die Basisschulung bietet eine gute Grundlage, auf der weitere Interventionen zur Verbesserung von Einstellungs- und Erkrankungsparametern aufbauen können. Sie eignet sich damit als zentraler Baustein für die rheumatologische Versorgung auf verschiedenen Ebenen. N2 - Background A new conceptual framework has enabled the flexible development of rheumatological patient educational programs for different healthcare settings. On this basis, a 5‑h basic training program for patients with rheumatoid arthritis (RA) was developed to be used in specialized centers. Rheumatologists and psychologists were first trained and then the efficacy of the patient training program was evaluated based on the causal model of patient education. Methods The externally randomized waiting control group study with 249 RA patients included 3 measurement points. The impact of the 5‑h basic training on disease and treatment-related knowledge as well as health competence of RA patients was examined. Secondary questions included attitudinal parameters, communication competence, effects on the disease and satisfaction with the educational program. Data were analyzed on an intention to treat basis by means of covariance analyses for the main target variables, adjusted for baseline values. Results The analyses showed that the training program was effective. Even 3 months after training, participants reported more knowledge and health competence than the waiting control group, with small to medium-sized effects (d = 0.37 and 0.38, respectively). With the exception of disease communication, no other effects of training were observed in the secondary objectives. Conclusion The basic training program provides a good foundation to develop further interventions to improve attitudinal and disease parameters. It can serve as a central component for rheumatological healthcare for patients with RA at various levels. KW - Rheumatoide Arthritis KW - Patientenschulung KW - Evaluation KW - Rheumatoid arthritis KW - Patient education KW - Evaluation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-280359 VL - 79 ER - TY - JOUR A1 - Hauer, Nadine N. A1 - Popp, Bernt A1 - Taher, Leila A1 - Vogl, Carina A1 - Dhandapany, Perundurai S. A1 - Büttner, Christian A1 - Uebe, Steffen A1 - Sticht, Heinrich A1 - Ferrazzi, Fulvia A1 - Ekici, Arif B. A1 - De Luca, Alessandro A1 - Klinger, Patrizia A1 - Kraus, Cornelia A1 - Zweier, Christiane A1 - Wiesener, Antje A1 - Abou Jamra, Rami A1 - Kunstmann, Erdmute A1 - Rauch, Anita A1 - Wieczorek, Dagmar A1 - Jung, Anna-Marie A1 - Rohrer, Tilman R. A1 - Zenker, Martin A1 - Doerr, Helmuth-Guenther A1 - Reis, André A1 - Thiel, Christian T. T1 - Evolutionary conserved networks of human height identify multiple Mendelian causes of short stature JF - European Journal of Human Genetics N2 - Height is a heritable and highly heterogeneous trait. Short stature affects 3% of the population and in most cases is genetic in origin. After excluding known causes, 67% of affected individuals remain without diagnosis. To identify novel candidate genes for short stature, we performed exome sequencing in 254 unrelated families with short stature of unknown cause and identified variants in 63 candidate genes in 92 (36%) independent families. Based on systematic characterization of variants and functional analysis including expression in chondrocytes, we classified 13 genes as strong candidates. Whereas variants in at least two families were detected for all 13 candidates, two genes had variants in 6 (UBR4) and 8 (LAMA5) families, respectively. To facilitate their characterization, we established a clustered network of 1025 known growth and short stature genes, which yielded 29 significantly enriched clusters, including skeletal system development, appendage development, metabolic processes, and ciliopathy. Eleven of the candidate genes mapped to 21 of these clusters, including CPZ, EDEM3, FBRS, IFT81, KCND1, PLXNA3, RASA3, SLC7A8, UBR4, USP45, and ZFHX3. Fifty additional growth-related candidates we identified await confirmation in other affected families. Our study identifies Mendelian forms of growth retardation as an important component of idiopathic short stature. KW - disease genetics KW - DNA sequencing KW - genetic counselling Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227899 VL - 27 ER - TY - JOUR A1 - Gilder, Stuart A. A1 - Wack, Michael A1 - Kaub, Leon A1 - Roud, Sophie C. A1 - Petersen, Nikolai A1 - Heinsen, Helmut A1 - Hillenbrand, Peter A1 - Milz, Stefan A1 - Schmitz, Chistoph T1 - Distribution of magnetic remanence carriers in the human brain JF - Scientific Reports N2 - That the human brain contains magnetite is well established; however, its spatial distribution in the brain has remained unknown. We present room temperature, remanent magnetization measurements on 822 specimens from seven dissected whole human brains in order to systematically map concentrations of magnetic remanence carriers. Median saturation remanent magnetizations from the cerebellum were approximately twice as high as those from the cerebral cortex in all seven cases (statistically significantly distinct, p = 0.016). Brain stems were over two times higher in magnetization on average than the cerebral cortex. The ventral (lowermost) horizontal layer of the cerebral cortex was consistently more magnetic than the average cerebral cortex in each of the seven studied cases. Although exceptions existed, the reproducible magnetization patterns lead us to conclude that magnetite is preferentially partitioned in the human brain, specifically in the cerebellum and brain stem. KW - brain KW - neurophysiology Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233035 VL - 8 ER - TY - JOUR A1 - Gotru, Sanjeev Kiran A1 - van Geffen, Johanna P. A1 - Nagy, Magdolna A1 - Mammadova-Bach, Elmina A1 - Eilenberger, Julia A1 - Volz, Julia A1 - Manukjan, Georgi A1 - Schulze, Harald A1 - Wagner, Leonard A1 - Eber, Stefan A1 - Schambeck, Christian A1 - Deppermann, Carsten A1 - Brouns, Sanne A1 - Nurden, Paquita A1 - Greinacher, Andreas A1 - Sachs, Ulrich A1 - Nieswandt, Bernhard A1 - Hermanns, Heike M. A1 - Heemskerk, Johan W. M. A1 - Braun, Attila T1 - Defective Zn2+ homeostasis in mouse and human platelets with α- and δ-storage pool diseases JF - Scientific Reports N2 - Zinc (Zn2+) can modulate platelet and coagulation activation pathways, including fibrin formation. Here, we studied the (patho)physiological consequences of abnormal platelet Zn2+ storage and release. To visualize Zn2+ storage in human and mouse platelets, the Zn2+ specific fluorescent dye FluoZin3 was used. In resting platelets, the dye transiently accumulated into distinct cytosolic puncta, which were lost upon platelet activation. Platelets isolated from Unc13d−/− mice, characterized by combined defects of α/δ granular release, showed a markedly impaired Zn2+ release upon activation. Platelets from Nbeal2−/− mice mimicking Gray platelet syndrome (GPS), characterized by primarily loss of the α-granule content, had strongly reduced Zn2+ levels, which was also confirmed in primary megakaryocytes. In human platelets isolated from patients with GPS, Hermansky-Pudlak Syndrome (HPS) and Storage Pool Disease (SPD) altered Zn2+ homeostasis was detected. In turbidity and flow based assays, platelet-dependent fibrin formation was impaired in both Nbeal2−/− and Unc13d−/− mice, and the impairment could be partially restored by extracellular Zn2+. Altogether, we conclude that the release of ionic Zn2+ store from secretory granules upon platelet activation contributes to the procoagulant role of Zn2+ in platelet-dependent fibrin formation. KW - coagulation system KW - metals Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227455 VL - 9 ER - TY - JOUR A1 - Gore, Lia A1 - Locatelli, Franco A1 - Zugmaier, Gerhard A1 - Handgretinger, Rupert A1 - O'Brien, Maureen M. A1 - Bader, Peter A1 - Bhojwani, Deepa A1 - Schlegel, Paul-Gerhardt A1 - Tuglus, Catherine A. A1 - Stackelberg, Arend von T1 - Survival after blinatumomab treatment in pediatric patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia JF - Blood Cancer Journal N2 - no abstract available KW - acute lymphocytic leukaemia KW - immunotherapy Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230726 VL - 8 ER - TY - JOUR A1 - Franchini, Paolo A1 - Jones, Julia C. A1 - Xiong, Peiwen A1 - Kneitz, Susanne A1 - Gompert, Zachariah A1 - Warren, Wesley C. A1 - Walter, Ronald B. A1 - Meyer, Axel A1 - Schartl, Manfred T1 - Long-term experimental hybridisation results in the evolution of a new sex chromosome in swordtail fish JF - Nature Communications N2 - The remarkable diversity of sex determination mechanisms known in fish may be fuelled by exceptionally high rates of sex chromosome turnovers or transitions. However, the evolutionary causes and genomic mechanisms underlying this variation and instability are yet to be understood. Here we report on an over 30-year evolutionary experiment in which we tested the genomic consequences of hybridisation and selection between two Xiphophorus fish species with different sex chromosome systems. We find that introgression and imposing selection for pigmentation phenotypes results in the retention of an unexpectedly large maternally derived genomic region. During the hybridisation process, the sex-determining region of the X chromosome from one parental species was translocated to an autosome in the hybrids leading to the evolution of a new sex chromosome. Our results highlight the complexity of factors contributing to patterns observed in hybrid genomes, and we experimentally demonstrate that hybridisation can catalyze rapid evolution of a new sex chromosome. KW - evolutionary genetics KW - experimental evolution KW - genome evolution Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228396 VL - 9 ER - TY - THES A1 - Stürzebecher, Paulina Elena T1 - Die Rolle von LASP1 in der Pathogenese der Atherosklerose im murinen Modell T1 - The role of LASP1 in the pathogenesis of atherosclerosis in mice N2 - Das regulatorische Gerüst-Protein LASP1, welches aus der Krebsforschung bekannt ist, wurde 2012 in humanen Makrophagen, den Protagonisten der Atherosklerose nachgewiesen. LASP1 ist durch seine Lokalisation an dynamischen Aktinskelettkonstruktionen (vgl. Invadopodien, Podosomen), nachweislich an Zellmigration, Proliferation und Invasionsfähigkeit bestimmter Tumorzellen beteiligt. Aufgrund einer großen Schnittmenge der Entstehungsmechanismen und zugrundeliegenden Signalwegen von Krebserkrankungen und Atherosklerose wurde LASP1 im Zusammenhang der Atherosklerose untersucht. In einem 16 Wochen Hochfettdiätversuch zeigten LASP1.Ldlr-/--Mäuse mehr atherosklerotische Läsionen in der Gesamtaorta als Ldlr-/--Tiere, was eine athero-protektive Rolle von LASP1 nahelegt. Passend hierzu führte Stimulation mit oxLDL in Makrophagen zu einer Hochregulation von LASP1. Zusätzlich internalisierten LASP1-/--Makrophagen signifikant mehr oxLDL im Vergleich zu LASP1-exprimierenden Zellen. Analog zu den Daten aus der Krebsforschung konnte eine reduzierte endotheliale Adhäsion sowie chemotaktische Migration von Ldlr.LASP1-/--Monozyten im Vergleich zu Ldlr-/-- Monozyten festgestellt werden. Dies ließe isoliert betrachtet eine pro-atherogene Rolle von LASP1 vermuten. Ein Nachweis von LASP1 im Zellkern von BMDMs konnte, zusätzlich zum fehlenden Shuttelproteinpartner ZO-2, nicht erbracht werden. Die Interaktion von LASP1 mit Transkriptionsfaktoren scheint daher unwahrscheinlich. Kongruent mit diesen Ergebnissen zeigte sich keine Veränderung der Transkription, der Proteinexpression sowie Sekretion von TNF! und ADAM17 durch den LASP1-KO. Insgesamt kommt LASP1 eine zweifellos komplexe Rolle in der Atherogenese zu. Die Ergebnisse der HFD-Versuche legen nahe, dass die primär anti-atherosklerotischen Einflüsse von LASP1 in vivo gegenüber den eher pro-atherosklerotischen Effekten des Proteins in vitro überwiegen. N2 - As of today, the regulatory scaffold protein LASP1 is mainly known from cancer research. Through its localization at dynamic actin skeletal constructs (cf. invadopodia, podosomes), LASP1 has been shown to be involved in cell migration, proliferation, and invasiveness of certain tumor cells. In 2012, LASP1 was detected in human macrophages, the protagonists of atherosclerosis. Because of a large intersection of mechanisms and signaling pathways of cancer and atherosclerosis, we further explored the role of LASP1 in the context of atherosclerosis. In a 16-week high-fat diet experiment, LASP1.Ldlr-/- mice showed more atherosclerotic lesions in the total aorta than Ldlr-/- animals, suggesting an athero-protective role of LASP1. In vitro, LASP1-/- macrophages internalized significantly more oxLDL than LASP1 positive cells. LASP1 did not shuttle in the nucleus of bone marrow derived macrophages. Thus, the interaction of LASP1 with transcription factors seems unlikely. Congruent with these results, LASP1-KO had no effects on the transcription, protein expression, or secretion of TNFα and ADAM17. In accordance with the data from cancer research, reduced endothelial adhesion as well as chemotactic migration of Ldlr.LASP1-/- monocytes was detected compared to Ldlr-/-- monocytes. These findings on the other hand would suggest a pro-atherogenic role of LASP1. Overall, LASP1 undoubtedly has a complex role in atherogenesis. The results of the HFD experiments suggest that the primarily anti-atherosclerotic influences of LASP1 in vivo predominate over the more pro-atherosclerotic effects of the protein in vitro. KW - Arteriosklerose KW - Schaumzelle KW - Maus KW - Monozyt KW - lasp Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239353 ER - TY - THES A1 - Friedrich, Anna-Lena T1 - FoxO3-mediated, inhibitory effects of CNP on the profibrotic activation of lung fibroblasts T1 - FoxO3-vermittelte, hemmende Wirkungen von CNP auf die profibrotische Aktivierung von Lungenfibroblasten N2 - Idiopathic Pulmonary Fibrosis (IPF) is a progressive parenchymal lung disease with limited therapeutic treatments. Pathologically altered lung fibroblasts, called myofibroblasts, exhibit increased proliferation, migration, and collagen production, and drive IPF development and progression. Fibrogenic factors such as Platelet derived growth factor-BB (PDGF-BB) contribute to these pathological alterations. Endogenous counter-regulating factors are barely known. Published studies have described a protective role of exogenously administered C-type Natriuretic Peptide (CNP) in pathological tissue remodeling, for example in heart and liver fibrosis. CNP and its cyclic GMP producing guanylyl cyclase B (GC-B) receptor are expressed in the lungs, but it is unknown whether CNP can attenuate lung fibrosis by this pathway. To address this question, we performed studies in primary cultured lung fibroblasts. To examine the effects of the CNP/GC-B pathway on PDGF-BB-induced collagen production, proliferation, and migration in vitro, lung fibroblasts were cultured from wildtype control and GC-B knockout mice. Human lung fibroblasts from patients with IPF and healthy controls were obtained from the UGMLC Biobank. In RIA experiments, CNP, at 10nM and 100nM, markedly and similarly increased cGMP levels in both the murine and human lung fibroblasts, demonstrating GC-B/cGMP signaling. CNP reduced PDGF-BB induced proliferation and migration of lung fibroblasts in BrdU incorporation and gap closure assays, respectively. CNP strongly decreased PDGF-BB-induced collagen 1/3 expression as measured by immunocytochemistry and immunoblotting. Importantly, the protective actions of CNP were preserved in IPF fibroblasts. It is known that the profibrotic actions of PDGF-BB are partly mediated by phosphorylation and nuclear export of Forkhead Box O3 (FoxO3), a transcription factor downregulated in IPF. CNP prevented PDGF-BB elicited FoxO3 phosphorylation and nuclear exclusion in both murine and human control and IPF fibroblasts. CNP signaling and functions were abolished in GC-B-deficient lung fibroblasts. Taken together, the results show that CNP moderates the PDGF-BB-induced activation and differentiation of human and murine lung fibroblasts to myofibroblasts. This effect is mediated CNP-dependent by GC-B/cGMP signaling and FoxO3 regulation. To follow up the patho-physiological relevance of these results, we are generating mice with fibroblast-restricted GC-B deletion for studies in the model of bleomycin-induced pulmonary fibrosis. N2 - Idiopathische pulmonale Fibrose (IPF) ist eine progressiv fortschreitende, parenchymale Lungenerkrankung mit beschränkten therapeutischen Behandlungsmöglichkeiten. Pathologisch veränderte Lungenfibroblasten, sogenannte Myofibroblasten, zeigen eine verstärkte Proliferation, Migration und Kollagenproduktion, die zu einem permanenten Fortschreiten der Erkrankung führen. Während fibrogene Faktoren wie Platelet derived growth factor-BB (PDGF-BB) zu dieser pathologischen Veränderung beitragen, sind endogene Faktoren, die diesem Wandel entgegenwirken, kaum bekannt. Allerdings konnten Studien bereits eine protektive Wirkung von exogen verabreichten C-typ natriuretischen Peptid (CNP) auf krankhaft verändertes Gewebe beschreiben, wie beispielsweise bei Herz- und Leberfibrose. Es ist bekannt, dass CNP und sein cGMP produzierender Rezeptor Guanylyl-Cyclase-B Rezeptor (GC-B) in der Lunge exprimiert werden. Allerdings konnte noch nicht nachgewiesen werden, ob CNP durch diesen Signalweg den Fortschritt einer Lungenfibrose verzögern kann. Um dies herauszufinden, wurden Experimente mit kultivierten, primären Lungenfibroblasten durchgeführt. Um die Effekte des CNP/ GC-B Signalwegs auf die PDGF-BB induzierte Kollagenproduktion, Proliferation und Migration in vitro zu überprüfen, wurden Lungenfibroblasten von Kontroll- und GC-B-Knock-Out Mäusen kultiviert. Weiterhin erhielten wir von der UGMLC Biobank menschliche Fibroblasten von IPF-erkrankten Patienten, sowie gesunde Kontrollfibroblasten. Zur Bestätigung der Funktionalität des GC-B/cGMP Weges, wurde in murinen und humanen Fibroblasten mithilfe eines RIAs ein durch CNP (10nM und 100nM) deutlich erhöhtes cGMP-Level gemessen. CNP reduzierte die durch PDGF-BB induzierte Beschleunigung von Proliferation und Migration der Lungenfibroblasten, was mit Hilfe von BrdU incorporation und Gap closure assay nachgewiesen wurde. Ebenfalls zeigten Immunocytochemistrie und -blotting, dass CNP den PDGF-BB induzierten Anstieg an Kollagenexpression verhindert. Somit wurde festgestellt, dass der schützende Effekt von CNP auch in IPF Fibroblasten erhalten bleibt. Weiterhin ist bekannt, dass die PDGF-BB-induzierten, profibrotischen Veränderungen durch Phosphorylierung und Export des Transkriptionsfaktors Forkhead Box O3 (FoxO3) aus dem Zellkern vermittelt werden, welcher in IPF Fibroblasten vermindert exprimiert ist. CNP verhindert diese PDGF-BB aktivierte Phosphorylierung und Translokation in murinen und humanen Kontroll- und IPF-Fibroblasten. In Lungenfibroblasten mit Deletion des GC-B- Rezeptors war das CNP-Signal und auch dessen Effekt ausgelöscht. Zusammengefasst zeigen die Ergebnisse, dass CNP die PDGF-BB induzierte Aktivierung und Differenzierung von menschlichen und murinen Lungenfibroblasten zu Myofibroblasten beeinflusst. Dieser Effekt wird CNP-abhängig durch den GC-B/cGMP Signalweg und durch die Regulierung von FoxO3 vermittelt. Um abschließend die pathophysiologische Relevanz dieser Erkenntnisse zu zeigen, werden zukünftig Mäuse mit einer fibroblasten-spezifischen Deletion des GC-B Rezeptors für Studien in Bleomycin-induzierter Lungenfibrose genutzt. KW - Idiopathische pulmonale Fibrose KW - Transkriptionsfaktor KW - Natriuretisches Hormon KW - C-type natriuretic peptide KW - FoxO3 Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359845 ER - TY - THES A1 - Menger, Kristina Rebekka T1 - Bauchlagerung für nicht-intubierte ARDS- und COVID-19 Patient/innen: eine systematische Übersichtsarbeit T1 - Prone Positioning for non-intubated ARDS and COVID-19 patients: a systematic review N2 - Die Bauchlagerung von intubierten ARDS-Patient/innen mit einer schlechten Oxygenierung wird laut Leitlinie seit mehreren Jahren als supportive Therapiemaßnahme empfohlen. Im Rahmen der COVID-19 Pandemie wurde nun erstmalig die Bauchlagerung auch bei hypoxämischen, nicht-intubierten Patient/innen untersucht. Diese Fragestellung wurde in der vorliegenden Arbeit mittels einer systematischen Übersichtsarbeit betrachtet. Aufgrund der aktuellen Pandemiesituation wurden neben ARDS-Patient/innen im Allgemeinen insbesondere COVID-19 Patient/innen mit einem akuten Lungenversagen als Subgruppe untersucht. Am 21.11.2020 wurde eine systematische Suche nach Studien in den Datenbanken MEDLINE, Cochrane COVID-19 Study Register und Living Overview of the Evidence platform durchgeführt. Die Ergebnisse wurden, wo möglich, in Form einer Meta-Analyse zusammengefasst, in Tabellen darstellt oder deskriptiv beschrieben. Das Risiko für Bias wurde jeweils für die eingeschlossenen kontrollierten Studien mittels ROBINS-I beurteilt. Die Vertrauenswürdigkeit der Evidenz der gesamten Arbeit wurde mit Hilfe des GRADE-Ansatzes untersucht. Insgesamt wurden 30 Studien eingeschlossen, davon 4 kontrollierte Studien, keine RCTs. In 3 der kontrollierten Studien wurde die Bauchlagerung bei COVID-19 Patient/innen untersucht, in einer bei Patient/innen mit einem anderweitig verursachten ARDS. Es ist unklar, ob die Bauchlagerung die Intubationsrate (RR = 0,92; 95% KI: 0,59 - 1,44; I² = 65%; sehr niedrige Vertrauenswürdigkeit der Evidenz), die Mortalität (RR = 0,55; 95% KI: 0,23 - 1,30; I² = 60%; sehr niedrige Vertrauenswürdigkeit der Evidenz) und die Wahrscheinlichkeit für eine Aufnahme auf die Intensivstation (RR = 0,94; 95% KI: 0,54 - 1,63; I2 = 71%; sehr niedrige Vertrauenswürdigkeit der Evidenz) verringern kann. Auch für die anderen betrachteten Endpunkte konnte kein signifikanter Effekt der Bauchlagerung nachgewiesen werden Im Vergleich der Subgruppen „Nicht-COVID-19“ (8 Studien) und „COVID-19“ (22 Studien) konnten in Bezug auf alle betrachteten Endpunkte keine relevanten Unterschiede festgestellt werden. Insgesamt ist die Evidenz nicht ausreichend, um Vor- und Nachteile der Bauchlagerung für nicht-intubierte ARDS Patient/innen gegenüber der üblichen Rückenlagerung aufzuzeigen und diese für die Praxis zu empfehlen. N2 - Prone positioning of intubated ARDS patients with poor oxygenation has been recommended as a supportive therapy for several years. In the context of the COVID-19 pandemic, prone positioning has now been investigated for the first time in hypoxemic, non-intubated patients. This issue was examined by our systematic review. Because of the current pandemic situation, we assessed the effects of prone positioning in COVID-19 patients with acute respiratory failure in particular as a subgroup in addition to ARDS patients in general. A systematic search for studies was performed in MEDLINE, Cochrane COVID-19 Study Register, and Living Overview of the Evidence platform databases on 21 November 2020. Where possible, results were analyzed in the form of meta-analysis, presented in tables, or described descriptively. Risk of bias was assessed for each of the included controlled studies using ROBINS-I. The quality of evidence for the entire systematic review was assessed using the GRADE approach. A total of 30 studies were included, 4 of which were controlled trials, but no RCTs. 3 of the controlled trials investigated prone positioning in COVID-19 patients, one trial in patients with ARDS caused by other means. It is unclear whether prone positioning can reduce intubation rate (RR =0.92; 95% CI: 0.59-1.44; I²=65%; very low quality evidence), mortality (RR =0.55; 95% CI: 0.23-1.30; I²=60%; very low quality evidence), and the likelihood of ICU admission (RR =0.94; 95% CI: 0.54-1.63; I2 =71%; very low quality evidence). No significant effect of prone positioning was demonstrated for the other outcomes considered either. When comparing the "non-COVID-19" (8 studies) and "COVID-19" (22 studies) subgroups, no relevant differences were found for all outcomes assessed. Overall, the evidence is insufficient to demonstrate benefits and harms of prone positioning for non-intubated ARDS patients compared with usual care and to translate the results into practice. KW - Bauchlage KW - ARDS KW - COVID-19 KW - Nicht-invasive Beatmung Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359865 ER - TY - THES A1 - Hajduk, Maurice Martin T1 - Darf es etwas mehr sein? Neuroenhancement im Studium – eine Befragung an Würzburger Hochschulen T1 - Can it be a little more? Neuroenhancement during studies - a survey at Würzburg universities N2 - Neuroenhancement (NE) bezeichnet die Einnahme psychotroper Substanzen mit dem Ziel der geistigen Leistungssteigerung oder Beruhigung. NE wird durch gesunde Perso- nen genutzt. Es besteht somit keine Indikation zur Einnahme psychotroper Wirkstoffe. Zum NE genutzte Substanzen sind z.B. Koffeintabletten, verschreibungspflichtige Medi- kamente oder illegale Substanzen. Die bisherige Forschung findet Hinweise auf einen Zusammenhang zwischen NE und ADHS-Symptomen, einigen Aspekten psychischer Gesundheit, sowie Substanzkonsum. Bisher gibt es keine Forschung zu NE am Hoch- schulstandort Würzburg. Es wurde eine anonyme online Querschnittsbefragung im ersten Quartal 2021 durchge- führt. Eingeladen waren 5600 Studierende der Julius-Maximilians-Universität Würzburg und der Hochschule für angewandte Wissenschaften Würzburg Schweinfurt. Der Frage- bogen bestand aus 53 Items und enthielt u. a. die folgenden validierten Messinstrumente: ASRS, PSS-10, PHQ-4 und AUDIT-C. Die Response Rate lag bei 18% (n = 1011). Das Wissen über NE war weit unter den Stu- dierenden verbreitet. Die Prävalenz für Neuroenhancement im Studium lag bei 12.7%. Die drei meistgenannten Substanzen waren Koffeintabletten (6.6%), Cannabis (4.5%) und Methylphenidat (4.3%). Häufigster Anlass für NE war die Prüfungsvorbereitung. Es zeigten sich deutliche Unterschiede zwischen den Fachbereichen, u.a. hinsichtlich der Prävalenz von NE. ADHS-Symptomen, Stress, Ängstlichkeit, und Depressivität waren positiv mit NE assoziiert. Ein stärkerer Effekt ergab sich für den Zusammenhang zwi- schen NE und riskanten Alkoholkonsum bzw. Tabakkonsum. Diese Ergebnisse wurden durch eine binomial logistische Regression bestätigt. Die konsumierten Substanzen, das Wissen über NE, die Prävalenz von NE und die Gründe für dessen Nutzung fügen sich nahtlos in die bisherige Forschung ein. Auch die Assoziation zwischen ADHS-Symptomen, Stress, Ängstlichkeit, Depressivität, riskan- tem Alkoholkonsum und Tabakkonsum bestätigt bisherige Forschungsergebnisse. Es konnte gezeigt werden, dass rund ein Zehntel der Studierenden NE bereits genutzt haben. In Anbetracht der gesundheitlichen Gefahren, die mit NE einhergehen ist die Etab- lierung bzw. der Ausbau von Aufklärung-, Beratungs- und Hilfsangeboten für Studie- rende anzustreben sowie weitere Forschung zum Thema indiziert. N2 - Neuroenhancement (NE) refers to the use of psychotropic substances with the aim of increasing mental performance or calming down. NE is used by healthy people. There is therefore no indication to take psychotropic substances. Substances used for NE include caffeine tablets, prescription drugs or illegal substances. Research has found evidence of a link between NE and ADHD symptoms, some aspects of mental health, and substance use. Up to now there has been no research on NE at the Würzburg university site. An anonymous online cross-sectional survey was conducted in the first quarter of 2021. 5,600 students from Julius-Maximilians-Universität Würzburg and the University of Applied Sciences Würzburg Schweinfurt were invited to participate. The questionnaire consisted of 53 items and included the following validated screening instruments: ASRS, PSS-10, PHQ-4 and AUDIT-C. The response rate was 18% (n = 1011). Knowledge about NE was widespread among the students. The prevalence of neuroenhancement during studies was 12.7%. The three most frequently mentioned substances were caffeine tablets (6.6%), cannabis (4.5%) and methylphenidate (4.3%). The most common reason for NE was exam preparation. There were clear differences between the subject areas, especially with regard to the prevalence of NE. ADHD symptoms, stress, anxiety and depression were positively associated with NE. A stronger effect was found for the relationship between NE and risky alcohol consumption or tobacco use. These results were confirmed by binomial logistic regression. The substances consumed, the knowledge about NE, the prevalence of NE and the reasons for its use fit in seamlessly with previous research. The association between ADHD symptoms, stress, anxiety, depression, risky alcohol consumption and tobacco consumption also confirms previous research findings. It was shown that around a tenth of students have already used NE. In view of the health risks associated with NE, the establishment or expansion of educational programs, counseling and support services for students is desirable and further research on the topic is indicated. KW - Psychische Gesundheit KW - Neuroenhancement KW - Cognitive Enhancement KW - Brain doping Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-359812 ER - TY - THES A1 - Hammel, Clara T1 - Einfluss longitudinaler Veränderungen der linksventrikulären Ejektionsfraktion auf das Langzeitüberleben bei Herzinsuffizienzpatienten mit leicht reduzierter Ejektionsfraktion oder reduzierter Ejektionsfraktion T1 - Impact of longitudinal changes in left ventricular ejection fraction on outcomes of patients with mid-range ejection fraction and reduced ejection N2 - Diese retrospektive Studie an der Universitätsklinik Würzburg diente der Beurteilung der longitudinalen Funktion in Bezug auf die Gesamtmortalität bei Patienten mit HFmrEF und HFrEF. Die Gruppierung erfolgte anhand der jeweiligen Baseline LVEF. Eine weitere Unterteilung erfolgte in eine ischämische oder nicht-ischämische Genese der HF. Die Subgruppen wurden anhand der Baseline klinischen Charakteristika sowie der echokardiographischen Parameter verglichen. Hier ließ sich ein relativ ähnliches Patientenklientel mit vergleichbarem Alter, Geschlecht, BMI sowie kardialen Risikofaktoren zeigen. Signifikante Unterschiede ergab der Vergleich des NYHA-Stadiums, der Nierenfunktion sowie des Auftretens von Myokardinfarkten. Die Veränderung der LVEF über die Zeit hat einen zentralen Stellenwert zur Evaluation des Outcomes von Patienten mit HFmrEF und HFrEF. Eine Verbesserung der LVEF fand sich signifikant häufiger bei HFrEF Patienten als bei HFmrEF Patienten, welche über die Zeit signifikant häufiger eine stabile LVEF aufwiesen. Außerdem war nach Auswertung der Überlebenskurven nach Kaplan-Meier in HFmrEF Patienten eine verbesserte oder unveränderte LVEF über die Zeit mit einem besseren Überleben verbunden, vor allem bei Patienten mit ischämischer Ätiologie. In der HFrEF Gruppe konnte gezeigt werden, dass sowohl Patienten mit ischämischer als auch mit nicht-ischämischer Ätiologie bei Vorliegen einer verbesserten oder unveränderten LVEF über die Zeit ein besseres Outcome aufwiesen. Eine erniedrigte MAPSE bedeutete vor allem bei HFmrEF Patienten mit nicht-ischämischer Ätiologie ein schlechteres Outcome. Die Ergebnisse dienten unter anderem der weiteren Charakterisierung der HFmrEF und HFrEF Gruppe sowie der Identifikation von Faktoren zur Beurteilung der Veränderung der LVEF über die Zeit und der Prognose des Langzeitüberlebens beider Gruppen. Ziel für die Zukunft sollte sein, auch für HFmrEF Patienten evidenzbasierte Herzinsuffizienz Therapien zu etablieren. N2 - This retrospective study at the University Hospital of Wuerzburg was designed to assess longitudinal function in relation to all-cause mortality in patients with HFmrEF and HFrEF. The grouping was based on the respective baseline LVEF. A further subdivision was made using coronary angiographic data into ischemic or non-ischemic genesis of HF. The subgroups were compared on the basis of baseline clinical characteristics and echocardiographic parameters. This revealed a relatively similar patient cohort with comparable age, gender, BMI and cardiac risk factors. Significant differences were found in the comparison of NYHA stage, renal function and the occurrence of myocardial infarction. The change in LVEF over time is of central importance for evaluating the outcome of patients with HFmrEF and HFrEF. An improvement in LVEF was significantly more common in HFrEF patients than in HFmrEF patients, who were significantly more likely to have a stable LVEF over time. Furthermore, according to the Kaplan-Meier survival curves in HFmrEF patients, improved or unchanged LVEF over time was associated with better survival, especially in patients with ischemic etiology. In the HFrEF group, it was shown that both patients with ischemic and non-ischemic etiology had a better outcome with improved or unchanged LVEF over time. A lower MAPSE was associated with a worse outcome, especially in HFmrEF patients with non-ischemic etiology. The results were used, among other things, to further characterize the HFmrEF and HFrEF groups and to identify factors for evaluating the change in LVEF over time and the prognosis of long-term survival in both groups. The aim for the future should be to establish evidence-based heart failure therapies for HFmrEF patients as well. KW - Transthorakale Echokardiographie KW - Herzinsuffizienz KW - MAPSE KW - HFmrEF KW - HFrEF KW - Langzeitüberleben KW - leicht reduzierte Herzinsuffizienz KW - reduzierte Herzinsuffizienz KW - longitudinale Funktion Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-360025 ER - TY - JOUR A1 - Allert, Stefanie A1 - Förster, Toni M. A1 - Svensson, Carl-Magnus A1 - Richardson, Jonathan P. A1 - Pawlik, Tony A1 - Hebecker, Betty A1 - Rudolphi, Sven A1 - Juraschitz, Marc A1 - Schaller, Martin A1 - Blagojevic, Mariana A1 - Morschhäuser, Joachim A1 - Figge, Marc Thilo A1 - Jacobsen, Ilse D. A1 - Naglik, Julian R. A1 - Kasper, Lydia A1 - Mogavero, Selene A1 - Hube, Bernhard T1 - \(Candida\) \(albicans\)-Induced Epithelial Damage Mediates Translocation through Intestinal Barriers JF - mBio N2 - Life-threatening systemic infections often occur due to the translocation of pathogens across the gut barrier and into the bloodstream. While the microbial and host mechanisms permitting bacterial gut translocation are well characterized, these mechanisms are still unclear for fungal pathogens such as Candida albicans, a leading cause of nosocomial fungal bloodstream infections. In this study, we dissected the cellular mechanisms of translocation of C. albicans across intestinal epithelia in vitro and identified fungal genes associated with this process. We show that fungal translocation is a dynamic process initiated by invasion and followed by cellular damage and loss of epithelial integrity. A screen of >2,000 C. albicans deletion mutants identified genes required for cellular damage of and translocation across enterocytes. Correlation analysis suggests that hypha formation, barrier damage above a minimum threshold level, and a decreased epithelial integrity are required for efficient fungal translocation. Translocation occurs predominantly via a transcellular route, which is associated with fungus-induced necrotic epithelial damage, but not apoptotic cell death. The cytolytic peptide toxin of C. albicans, candidalysin, was found to be essential for damage of enterocytes and was a key factor in subsequent fungal translocation, suggesting that transcellular translocation of C. albicans through intestinal layers is mediated by candidalysin. However, fungal invasion and low-level translocation can also occur via non-transcellular routes in a candidalysin-independent manner. This is the first study showing translocation of a human-pathogenic fungus across the intestinal barrier being mediated by a peptide toxin. IMPORTANCE Candida albicans, usually a harmless fungus colonizing human mucosae, can cause lethal bloodstream infections when it manages to translocate across the intestinal epithelium. This can result from antibiotic treatment, immune dysfunction, or intestinal damage (e.g., during surgery). However, fungal processes may also contribute. In this study, we investigated the translocation process of C. albicans using in vitro cell culture models. Translocation occurs as a stepwise process starting with invasion, followed by epithelial damage and loss of epithelial integrity. The ability to secrete candidalysin, a peptide toxin deriving from the hyphal protein Ece1, is key: C. albicans hyphae, secreting candidalysin, take advantage of a necrotic weakened epithelium to translocate through the intestinal layer. KW - Candida albicans KW - candidalysin KW - host cell damage KW - host cell invasion KW - intestinal barrier KW - necrosis KW - translocation Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-221084 VL - 9 IS - 3 ER -