TY - JOUR A1 - Ziegler, Sabrina A1 - Weiss, Esther A1 - Schmitt, Anna-Lena A1 - Schlegel, Jan A1 - Burgert, Anne A1 - Terpitz, Ulrich A1 - Sauer, Markus A1 - Moretta, Lorenzo A1 - Sivori, Simona A1 - Leonhardt, Ines A1 - Kurzai, Oliver A1 - Einsele, Hermann A1 - Loeffler, Juergen T1 - CD56 Is a Pathogen Recognition Receptor on Human Natural Killer Cells JF - Scientific Reports N2 - Aspergillus (A.) fumigatus is an opportunistic fungal mold inducing invasive aspergillosis (IA) in immunocompromised patients. Although antifungal activity of human natural killer (NK) cells was shown in previous studies, the underlying cellular mechanisms and pathogen recognition receptors (PRRs) are still unknown. Using flow cytometry we were able to show that the fluorescence positivity of the surface receptor CD56 significantly decreased upon fungal contact. To visualize the interaction site of NK cells and A. fumigatus we used SEM, CLSM and dSTORM techniques, which clearly demonstrated that NK cells directly interact with A. fumigatus via CD56 and that CD56 is re-organized and accumulated at this interaction site time-dependently. The inhibition of the cytoskeleton showed that the receptor re-organization was an active process dependent on actin re-arrangements. Furthermore, we could show that CD56 plays a role in the fungus mediated NK cell activation, since blocking of CD56 surface receptor reduced fungal mediated NK cell activation and reduced cytokine secretion. These results confirmed the direct interaction of NK cells and A. fumigatus, leading to the conclusion that CD56 is a pathogen recognition receptor. These findings give new insights into the functional role of CD56 in the pathogen recognition during the innate immune response. KW - pattern recognition receptors KW - fungal infection KW - Aspergillus fumigatus KW - natural killer cells Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170637 VL - 7 IS - 6138 ER - TY - JOUR A1 - Frey, Anna A1 - Popp, Sandy A1 - Post, Antonia A1 - Langer, Simon A1 - Lehmann, Marc A1 - Hofmann, Ulrich A1 - Siren, Anna-Leena A1 - Hommers, Leif A1 - Schmitt, Angelika A1 - Strekalova, Tatyana A1 - Ertl, Georg A1 - Lesch, Klaus-Peter A1 - Frantz, Stefan T1 - Experimental heart failure causes depression-like behavior together with differential regulation of inflammatory and structural genes in the brain JF - Frontiers in Behavioral Neuroscience N2 - Background: Depression and anxiety are common and independent outcome predictors in patients with chronic heart failure (CHF). However, it is unclear whether CHF causes depression. Thus, we investigated whether mice develop anxiety- and depression-like behavior after induction of ischemic CHF by myocardial infarction (MI). Methods and Results: In order to assess depression-like behavior, anhedonia was investigated by repeatedly testing sucrose preference for 8 weeks after coronary artery ligation or sham operation. Mice with large MI and increased left ventricular dimensions on echocardiography (termed CHF mice) showed reduced preference for sucrose, indicating depression-like behavior. 6 weeks after MI, mice were tested for exploratory activity, anxiety-like behavior and cognitive function using the elevated plus maze (EPM), light-dark box (LDB), open field (OF), and object recognition (OR) tests. In the EPM and OF, CHF mice exhibited diminished exploratory behavior and motivation despite similar movement capability. In the OR, CHF mice had reduced preference for novelty and impaired short-term memory. On histology, CHF mice had unaltered overall cerebral morphology. However, analysis of gene expression by RNA-sequencing in prefrontal cortical, hippocampal, and left ventricular tissue revealed changes in genes related to inflammation and cofactors of neuronal signal transduction in CHF mice, with Nr4a1 being dysregulated both in prefrontal cortex and myocardium after MI. Conclusions: After induction of ischemic CHF, mice exhibited anhedonic behavior, decreased exploratory activity and interest in novelty, and cognitive impairment. Thus, ischemic CHF leads to distinct behavioral changes in mice analogous to symptoms observed in humans with CHF and comorbid depression. KW - chronic heart failure KW - myocardial infarction KW - anxiety KW - depression KW - mice Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-118234 SN - 1662-5153 VL - 8 ER - TY - JOUR A1 - Schul, Daniela A1 - Schmitt, Alexandra A1 - Regneri, Janine A1 - Schartl, Manfred A1 - Wagner, Toni Ulrich T1 - Bursted BMP Triggered Receptor Kinase Activity Drives Smad1 Mediated Long-Term Target Gene Oscillation in c2c12 Cells JF - PLoS ONE N2 - Bone Morphogenetic Proteins (BMPs) are important growth factors that regulate many cellular processes. During embryogenesis they act as morphogens and play a critical role during organ development. They influence cell fates via concentration-gradients in the embryos where cells transduce this extracellular information into gene expression profiles and cell fate decisions. How receiving cells decode and quantify BMP2/4 signals is hardly understood. There is little data on the quantitative relationships between signal input, transducing molecules, their states and location, and ultimately their ability to integrate graded systemic inputs and generate qualitative responses. Understanding this signaling network on a quantitative level should be considered a prerequisite for efficient pathway modulation, as the BMP pathway is a prime target for therapeutic invention. Hence, we quantified the spatial distribution of the main signal transducer of the BMP2/4 pathway in response to different types and levels of stimuli in c2c12 cells. We found that the subcellular localization of Smad1 is independent of ligand concentration. In contrast, Smad1 phosphorylation levels relate proportionally to BMP2 ligand concentrations and they are entirely located in the nucleus. Interestingly, we found that BMP2 stimulates target gene expression in non-linear, wave-like forms. Amplitudes showed a clear concentration-dependency, for sustained and transient stimulation. We found that even burst-stimulation triggers gene-expression wave-like modulations that are detectable for at least 30 h. Finally, we show here that target gene expression oscillations depend on receptor kinase activity, as the kinase drives further expression pulses without receptor reactivation and the target gene expression breaks off after inhibitor treatment in c2c12 cells. KW - gene expression KW - BMP signaling KW - SMAD signaling KW - genetic oscillators KW - cell fusion KW - DNA-binding proteins KW - luciferase KW - kinase inhibitors Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130131 VL - 8 IS - 4 ER - TY - JOUR A1 - Frey, Anna A1 - Gassenmaier, Tobias A1 - Hofmann, Ulrich A1 - Schmitt, Dominik A1 - Fette, Georg A1 - Marx, Almuth A1 - Heterich, Sabine A1 - Boivin-Jahns, Valérie A1 - Ertl, Georg A1 - Bley, Thorsten A1 - Frantz, Stefan A1 - Jahns, Roland A1 - Störk, Stefan T1 - Coagulation factor XIII activity predicts left ventricular remodelling after acute myocardial infarction JF - ESC Heart Failure N2 - Aims Acute myocardial infarction (MI) is the major cause of chronic heart failure. The activity of blood coagulation factor XIII (FXIIIa) plays an important role in rodents as a healing factor after MI, whereas its role in healing and remodelling processes in humans remains unclear. We prospectively evaluated the relevance of FXIIIa after acute MI as a potential early prognostic marker for adequate healing. Methods and results This monocentric prospective cohort study investigated cardiac remodelling in patients with ST-elevation MI and followed them up for 1 year. Serum FXIIIa was serially assessed during the first 9 days after MI and after 2, 6, and 12 months. Cardiac magnetic resonance imaging was performed within 4 days after MI (Scan 1), after 7 to 9 days (Scan 2), and after 12 months (Scan 3). The FXIII valine-to-leucine (V34L) single-nucleotide polymorphism rs5985 was genotyped. One hundred forty-six patients were investigated (mean age 58 ± 11 years, 13% women). Median FXIIIa was 118 % (quartiles, 102–132%) and dropped to a trough on the second day after MI: 109%(98–109%; P < 0.001). FXIIIa recovered slowly over time, reaching the baseline level after 2 to 6 months and surpassed baseline levels only after 12 months: 124 % (110–142%). The development of FXIIIa after MI was independent of the genotype. FXIIIa on Day 2 was strongly and inversely associated with the relative size of MI in Scan 1 (Spearman’s ρ = –0.31; P = 0.01) and Scan 3 (ρ = –0.39; P < 0.01) and positively associated with left ventricular ejection fraction: ρ = 0.32 (P < 0.01) and ρ = 0.24 (P = 0.04), respectively. Conclusions FXIII activity after MI is highly dynamic, exhibiting a significant decline in the early healing period, with reconstitution 6 months later. Depressed FXIIIa early after MI predicted a greater size of MI and lower left ventricular ejection fraction after 1 year. The clinical relevance of these findings awaits to be tested in a randomized trial. KW - blood coagulation factor XIII KW - ST-elevation myocardial infarction KW - healing and remodelling processes KW - cardiac magnetic resonance imaging Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-236013 VL - 7 IS - 5 ER - TY - THES A1 - Schmitt, Ulrich T1 - Cyclodextrine als chirale Selektoren in der kapillarelektrophoretischen Enantiomerentrennung T1 - Cyclodextrins as chiral selectors in capillary electrophoretic separation of enaantiomers N2 - Die vielfältigen isomeren Substanzen, die in dieser Arbeit getrennt wurden, zeigen, dass die cyclodextrin-modifizierte Kapillarelektrophorese eine sehr leistungsfähige Technik für die Trennung von Enantiomeren darstellt. Als kritisch erweist sich jedoch das Auffinden des richtigen Cyclodextrins bzw. dessen optimale Konzentration. Die Vorhersagbarkeit dieser Parameter ist noch zu gering, um hier vor den ersten Versuchen alleine anhand der Struktur des Moleküls sichere Aussagen machen zu können. So sind zur Zeit noch Versuchsreihen mit verschiedenen Cyclodextrinen und Bedingungen nötig, wie sie in dieser Arbeit durchgeführt wurden. Für die Zukunft wäre es wünschenswert, durch das Sammeln von mehr Erfahrungen und Ergebnissen, wie sie hier gezeigt wurden, schon möglichst direkt aus den Strukturen der zu trennenden Zielmoleküle eine Aussage über die Beschaffenheit des Cyclodextrins oder der CE-Methode machen zu können. Von sechs verschiedenen schwach basischen Thiobarbituratracematen konnten fünf erfolgreich in ihr Enantiomeren getrennt werden. Unter den fünf erfolgreich getrennt Barbituraten waren zwei am Stickstoff alkyliert und ein weiteres sowohl am Stickstoff als auch am Schwefel. Die Thiobarbiturate, die am Stickstoffatom keinen Substituenten tragen, konnten mit -CD am besten getrennt werden (RS > 6). Hingegen konnte für die beiden nur am Stickstoff alkylierten Barbituratemit mit -CD und HDMS jeweils Auflösungen über RS = 6 erzielt werden. Für den in der Erdbeere vorhandene Schlüsselaromastoff 2,5-Dimethyl-4-hydroxy-3(2H)-furanon, Furaneol, konnte eine CE-Methode entwickelt werden, die es zum einen ermöglicht, die Enantiomere vollständig zu trennen, und zum anderen so beschaffen ist, dass es bei der Trennung selbst zu keiner Racemisierung kommt, wie es bei der bisher verwendeten Gaschromatographie der Fall war. Die CE-Methode machte es erstmalig möglich, die enantioselektive Biosynthese von Furaneol in Erdbeerproteinextrakten nachzuweisen. Zusätzlich wurde noch die Racemisierungsgeschwindigkeit von einem angereichertem Gemisch bei vier verschiedenen pH-Werte getestet, was für weitere Arbeiten mit Furaneol-Proben von Bedeutung ist. Die Racemisierungsgeschwindigkeit erwies sich für den pH-Bereich von 3.8 bis 5 am langsamsten. Im neutralen Bereich wurde die schnellste Racemisierung beobachtet. Bei den Experimenten mit Atropin und dem verwandten Homatropin sollte geprüft werden, ob der Einsatz sulfatierter Cyclodextrine von vier verschiedenen Herstellern zu identischen Ergebnissen führt. Es konnten anhand einer kritischen Trennung gezeigt werden, dass dies nicht der Fall ist. Die Trennleistung der Cyclodextrine in Bezug auf den Vergleich Atropin/Homatropin war hierbei sehr ähnlich. Ein bei diesen Arbeiten zusätzlich entstandener Peak konnte eindeutig einem Zerfallsprodukt des Atropins, der Tropasäure, zugeordnet werden. Mit Hilfe von Experimenten mit den Atropisomeren von 1,1′-Binaphthalin-2,2′-diyl-phosphat sollte erprobt werden, ob eine Charakterisierung von randomisiert substituierter CDs über den durchschnittlichen molekularen Substitutionsgrad (MS) ausreicht, um beim erneuten Einsatz eines vergleichbaren Produktes reproduzierbare Ergebnisse zu bekommen. Die Trennleistung von unterschiedlich randomisiert substituierten acetylierten und methylierten Cyclodextrinen wurde hierzu getestet, und mit der von -CD und Heptakis-(2,3,6-tri-O-methyl)--cyclodextrin, bzw. deren Gemischen in Verhältnis von 1:9 bis 9:1 verglichen. Der Vergleich dieser Messergebnisse führte zu dem Schluss, dass eine einfache Charakterisierung randomisiert substituierter Cyclodextrine über den durchschnittlichen molekularen Substitutionsgrad (MS) nicht ausreicht, um reproduzierbare Ergebnisse beim Einsatz solcher Cyclodextrine zu erreichen. Die Aufnahme von Massenspektren ermöglichte es hingegen, randomisiert substituierte CDs aussagekräftig zu charakterisieren. Will man eine CE-Methode validieren, in der ein randomisiertes Cyclodextrin zu Einsatz kommt, so sollte man dieses mittels Massenspektroskopie charakterisieren und sich dessen bewusst sein, das die Validierung nur für diese eine Charge des Herstellers bzw. für Chargen mit vergleichbaren Massenspektren Gültigkeit besitzt. Für das im Arbeitskreis Bringmann racemisch synthetisierte 2-Hydroxymethyl-1-(2-hydroxy-4,6-dimethoxyphenyl)naphthalin sollte eine Trennmethode für die Atropisomere entwickelt werden, da für die Zukunft eine enantioselektive Synthese geplant ist. Unter Einsatz der chiralen Selektoren Heptakis-(6-sulfato)--CD (HS) und Heptakis-(2,3-di-O-acetyl-6-sulfato)--cyclodextrin (HDAS) konnten zwei Trennmethoden gefunden werden, die bei einer jeweiligen Cyclodextrinkonzentration von 15 mM zu einer guten Trennung der Enantiomeren führte. Dies ermöglicht eine Quantifizierung des Enantiomerenüberschuß (ee) für eine nicht-racemische Syntheseroute. N2 - The manifold of isomeric substances being separated within the frame of this thesis shows the power of cyclodextrin-modified capillary electrophoresis concerning to the separation of enantiomers. The major problem is still to find the appropriate cyclodextrin respectively the optimum concentration. The possibility to predict these parameters only on the basis of the molecule structure is still too small. Thus, it is still necessary, to perform experiments to find the suitable cyclodextrin. It would be desirable for the future, to make this possible by collecting more experiences and information. The enantiomers of five of six weakly basic thiobarbiturates could be successfully separated. Two of these five barbiturates were N-substituted, one N-substituted as well as S-substituted. The three N-substituted thiobarbiturates characterized by phenyl- and ethyl-substituent in five positions being the chiral centre. In the two other molecules the chiral centre is located in the side chain, bound at the five position. Furthermore the dependence of the cyclodextrin concentration on the resolution power of five commercial cyclodextrins could be shown. The concentration leading to the best resolution was varying from the smallest (1 mmol/l) to the highest (18 mmol/l) concentrations which were used. The thiobarbiturates without a substituent at the nitrogen atom could be separated best by using -CD (RS > 6). Whereas the two barbiturates carrying a N-alkyl substituent reach their best resolution with -CD and HDMS (both RS > 6). Beside the absolute maxima local maxima were observed. For the key flavouring substance of the strawberry, 2,5-dimethyl-4-hydroxy-3(2H)-furanon, furaneol, a CE-method was developed, which makes it possible to separate the enantiomers without racemisation, which happens when using gas chromatography. This makes it possible to verify enantioselective biosynthesis of furaneol in strawberry protein extract for the first time. Additional the rate of racemisation of enantiomeric enriched samples at four different pH-values were tested, which is important for further experiments with furaneol. The studies on atropine and the related homatropine were performed to show, whether four sulphated cyclodextrins of four different manufacturers lead to identical results. By means of a critical separation the differences of the resolution powers could be shown. It varies from no resolution over separation 1/3 of the peak to 2/3. The resolution power to the atropine enantiomeres was comparable to the one of homatropine. An additional peak, which was found in the tests, was assigned to a decomposition product of atropine being tropic acid. By means of a study with the atrop-isomers of 1,1′-binaphthalin-2,2′-diyl-phosphate it was checked, whether the characterisation of randomly substituted cyclodextrins via the average molecular degree of substitution (MS) is sufficient to obtain comparable results in CD-modified CE. The resolution power of different randomly substituted acetylated and methylated cyclodextrins were tested and compared to the one of -CD and heptakis-(2,3,6-tri-O-methyl)--cyclodextrin and their mixtures in a ratio of 1:9 to 9:1. The comparison of these results shows, that a simple classification of randomly substituted cyclodextrins by the average molecular degree of substitution (MS) is not sufficient to reproduce the results of measurements done with such kind of cyclodextrins. To validate a CE-method, in which a randomly substituted cyclodextrin is used, a mass spectrum of this cyclodextrin should be recorded and it should be clear, that the validation is effective only for this batch of the producer or for batches which have comparable mass spectra. For 2-hydroxymethyl-1-(2-hydroxy-4,6-dimethoxyphenyl)naphthalin, which was racemically synthesized in the group of Prof. Bringmann, a separation method for the atop-isomers was developed, because a enantioselective synthesis is planed in the future. Using either heptakis-(6-sulfato)--CD (HS) or heptakis-(2,3-di-O-acetyl-6-sulfato)--cyclodextrin (HDAS) as chiral selectors at a cyclodextrin concentration of 15 mM a good separation was obtained. This makes a quantification of the enantiomeric excess (ee) for enantioselective synthesis routes possible. KW - Enantiomerentrennung KW - Kapillarelektrophorese KW - Cyclodextrine KW - Kapillarelektrophorese KW - Cyclodextrin KW - Enantiomere KW - capillary electrophoresis KW - cyclodextrin KW - enantiomer Y1 - 2004 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-10043 ER - TY - JOUR A1 - Elias, Johannes A1 - Heuschmann, Peter U. A1 - Schmitt, Corinna A1 - Eckhardt, Frithjof A1 - Boehm, Hartmut A1 - Maier, Sebastian A1 - Kolb-Mäurer, Annette A1 - Riedmiller, Hubertus A1 - Müllges, Wolfgang A1 - Weisser, Christoph A1 - Wunder, Christian A1 - Frosch, Matthias A1 - Vogel, Ulrich T1 - Prevalence dependent calibration of a predictive model for nasal carriage of methicillin-resistant Staphylococcus aureus JF - BMC Infectious Diseases N2 - Background Published models predicting nasal colonization with Methicillin-resistant Staphylococcus aureus among hospital admissions predominantly focus on separation of carriers from non-carriers and are frequently evaluated using measures of discrimination. In contrast, accurate estimation of carriage probability, which may inform decisions regarding treatment and infection control, is rarely assessed. Furthermore, no published models adjust for MRSA prevalence. Methods Using logistic regression, a scoring system (values from 0 to 200) predicting nasal carriage of MRSA was created using a derivation cohort of 3091 individuals admitted to a European tertiary referral center between July 2007 and March 2008. The expected positive predictive value of a rapid diagnostic test (GeneOhm, Becton & Dickinson Co.) was modeled using non-linear regression according to score. Models were validated on a second cohort from the same hospital consisting of 2043 patients admitted between August 2008 and January 2012. Our suggested correction score for prevalence was proportional to the log-transformed odds ratio between cohorts. Calibration before and after correction, i.e. accurate classification into arbitrary strata, was assessed with the Hosmer-Lemeshow-Test. Results Treating culture as reference, the rapid diagnostic test had positive predictive values of 64.8% and 54.0% in derivation and internal validation corhorts with prevalences of 2.3% and 1.7%, respectively. In addition to low prevalence, low positive predictive values were due to high proportion (> 66%) of mecA-negative Staphylococcus aureus among false positive results. Age, nursing home residence, admission through the medical emergency department, and ICD-10-GM admission diagnoses starting with “A” or “J” were associated with MRSA carriage and were thus included in the scoring system, which showed good calibration in predicting probability of carriage and the rapid diagnostic test’s expected positive predictive value. Calibration for both probability of carriage and expected positive predictive value in the internal validation cohort was improved by applying the correction score. Conclusions Given a set of patient parameters, the presented models accurately predict a) probability of nasal carriage of MRSA and b) a rapid diagnostic test’s expected positive predictive value. While the former can inform decisions regarding empiric antibiotic treatment and infection control, the latter can influence choice of screening method. KW - Methicillin-resistant staphylococcus aureus KW - Infection control KW - Clinical prediction rule KW - Predictive value of tests KW - False positive reactions KW - Calibration Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-96091 UR - http://www.biomedcentral.com/1471-2334/13/111 ER - TY - JOUR A1 - Dischinger, Ulrich A1 - Heckel, Tobias A1 - Bischler, Thorsten A1 - Hasinger, Julia A1 - Königsrainer, Malina A1 - Schmitt-Böhrer, Angelika A1 - Otto, Christoph A1 - Fassnacht, Martin A1 - Seyfried, Florian A1 - Hankir, Mohammed Khair T1 - Roux-en-Y gastric bypass and caloric restriction but not gut hormone-based treatments profoundly impact the hypothalamic transcriptome in obese rats JF - Nutrients N2 - Background: The hypothalamus is an important brain region for the regulation of energy balance. Roux-en-Y gastric bypass (RYGB) surgery and gut hormone-based treatments are known to reduce body weight, but their effects on hypothalamic gene expression and signaling pathways are poorly studied. Methods: Diet-induced obese male Wistar rats were randomized into the following groups: RYGB, sham operation, sham + body weight-matched (BWM) to the RYGB group, osmotic minipump delivering PYY3-36 (0.1 mg/kg/day), liraglutide s.c. (0.4 mg/kg/day), PYY3-36 + liraglutide, and saline. All groups (except BWM) were kept on a free choice of high- and low-fat diets. Four weeks after interventions, hypothalami were collected for RNA sequencing. Results: While rats in the RYGB, BWM, and PYY3-36 + liraglutide groups had comparable reductions in body weight, only RYGB and BWM treatment had a major impact on hypothalamic gene expression. In these groups, hypothalamic leptin receptor expression as well as the JAK–STAT, PI3K-Akt, and AMPK signaling pathways were upregulated. No significant changes could be detected in PYY3-36 + liraglutide-, liraglutide-, and PYY-treated groups. Conclusions: Despite causing similar body weight changes compared to RYGB and BWM, PYY3-36 + liraglutide treatment does not impact hypothalamic gene expression. Whether this striking difference is favorable or unfavorable to metabolic health in the long term requires further investigation. KW - obesity KW - Roux-en-Y gastric bypass surgery KW - liraglutide KW - PYY3-36 KW - hypothalamic gene expression Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-252392 SN - 2072-6643 VL - 14 IS - 1 ER - TY - JOUR A1 - Popp, Sandy A1 - Schmitt-Böhrer, Angelika A1 - Langer, Simon A1 - Hofmann, Ulrich A1 - Hommers, Leif A1 - Schuh, Kai A1 - Frantz, Stefan A1 - Lesch, Klaus-Peter A1 - Frey, Anna T1 - 5-HTT Deficiency in Male Mice Affects Healing and Behavior after Myocardial Infarction JF - Journal of Clinical Medicine N2 - Anxiety disorders and depression are common comorbidities in cardiac patients. Mice lacking the serotonin transporter (5-HTT) exhibit increased anxiety-like behavior. However, the role of 5-HTT deficiency on cardiac aging, and on healing and remodeling processes after myocardial infarction (MI), remains unclear. Cardiological evaluation of experimentally naïve male mice revealed a mild cardiac dysfunction in ≥4-month-old 5-HTT knockout (−/−) animals. Following induction of chronic cardiac dysfunction (CCD) by MI vs. sham operation 5-HTT−/− mice with infarct sizes >30% experienced 100% mortality, while 50% of 5-HTT+/− and 37% of 5-HTT+/+ animals with large MI survived the 8-week observation period. Surviving (sham and MI < 30%) 5-HTT−/− mutants displayed reduced exploratory activity and increased anxiety-like behavior in different approach-avoidance tasks. However, CCD failed to provoke a depressive-like behavioral response in either 5-Htt genotype. Mechanistic analyses were performed on mice 3 days post-MI. Electrocardiography, histology and FACS of inflammatory cells revealed no abnormalities. However, gene expression of inflammation-related cytokines (TGF-β, TNF-α, IL-6) and MMP-2, a protein involved in the breakdown of extracellular matrix, was significantly increased in 5-HTT−/− mice after MI. This study shows that 5-HTT deficiency leads to age-dependent cardiac dysfunction and disrupted early healing after MI probably due to alterations of inflammatory processes in mice. KW - chronic heart failure KW - myocardial infarction KW - serotonin transporter deficient mice KW - anxiety KW - depression KW - behavior KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242739 SN - 2077-0383 VL - 10 IS - 14 ER - TY - JOUR A1 - Haeusner, Sebastian A1 - Herbst, Laura A1 - Bittorf, Patrick A1 - Schwarz, Thomas A1 - Henze, Chris A1 - Mauermann, Marc A1 - Ochs, Jelena A1 - Schmitt, Robert A1 - Blache, Ulrich A1 - Wixmerten, Anke A1 - Miot, Sylvie A1 - Martin, Ivan A1 - Pullig, Oliver T1 - From Single Batch to Mass Production–Automated Platform Design Concept for a Phase II Clinical Trial Tissue Engineered Cartilage Product JF - Frontiers in Medicine N2 - Advanced Therapy Medicinal Products (ATMP) provide promising treatment options particularly for unmet clinical needs, such as progressive and chronic diseases where currently no satisfying treatment exists. Especially from the ATMP subclass of Tissue Engineered Products (TEPs), only a few have yet been translated from an academic setting to clinic and beyond. A reason for low numbers of TEPs in current clinical trials and one main key hurdle for TEPs is the cost and labor-intensive manufacturing process. Manual production steps require experienced personnel, are challenging to standardize and to scale up. Automated manufacturing has the potential to overcome these challenges, toward an increasing cost-effectiveness. One major obstacle for automation is the control and risk prevention of cross contaminations, especially when handling parallel production lines of different patient material. These critical steps necessitate validated effective and efficient cleaning procedures in an automated system. In this perspective, possible technologies, concepts and solutions to existing ATMP manufacturing hurdles are discussed on the example of a late clinical phase II trial TEP. In compliance to Good Manufacturing Practice (GMP) guidelines, we propose a dual arm robot based isolator approach. Our novel concept enables complete process automation for adherent cell culture, and the translation of all manual process steps with standard laboratory equipment. Moreover, we discuss novel solutions for automated cleaning, without the need for human intervention. Consequently, our automation concept offers the unique chance to scale up production while becoming more cost-effective, which will ultimately increase TEP availability to a broader number of patients. KW - ATMP KW - tissue engineering KW - GMP KW - manufacturing KW - autologous KW - cartilage regeneration KW - automation & robotics KW - automation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244631 SN - 2296-858X VL - 8 ER - TY - JOUR A1 - Traub, Jan A1 - Grondey, Katja A1 - Gassenmaier, Tobias A1 - Schmitt, Dominik A1 - Fette, Georg A1 - Frantz, Stefan A1 - Boivin-Jahns, Valérie A1 - Jahns, Roland A1 - Störk, Stefan A1 - Stoll, Guido A1 - Reiter, Theresa A1 - Hofmann, Ulrich A1 - Weber, Martin S. A1 - Frey, Anna T1 - Sustained increase in serum glial fibrillary acidic protein after first ST-elevation myocardial infarction JF - International Journal of Molecular Sciences N2 - Acute ischemic cardiac injury predisposes one to cognitive impairment, dementia, and depression. Pathophysiologically, recent positron emission tomography data suggest astroglial activation after experimental myocardial infarction (MI). We analyzed peripheral surrogate markers of glial (and neuronal) damage serially within 12 months after the first ST-elevation MI (STEMI). Serum levels of glial fibrillary acidic protein (GFAP) and neurofilament light chain (NfL) were quantified using ultra-sensitive molecular immunoassays. Sufficient biomaterial was available from 45 STEMI patients (aged 28 to 78 years, median 56 years, 11% female). The median (quartiles) of GFAP was 63.8 (47.0, 89.9) pg/mL and of NfL 10.6 (7.2, 14.8) pg/mL at study entry 0–4 days after STEMI. GFAP after STEMI increased in the first 3 months, with a median change of +7.8 (0.4, 19.4) pg/mL (p = 0.007). It remained elevated without further relevant increases after 6 months (+11.7 (0.6, 23.5) pg/mL; p = 0.015), and 12 months (+10.3 (1.5, 22.7) pg/mL; p = 0.010) compared to the baseline. Larger relative infarction size was associated with a higher increase in GFAP (ρ = 0.41; p = 0.009). In contrast, NfL remained unaltered in the course of one year. Our findings support the idea of central nervous system involvement after MI, with GFAP as a potential peripheral biomarker of chronic glial damage as one pathophysiologic pathway. KW - myocardial infarction KW - STEMI KW - glial fibrillary acidic protein KW - GFAP KW - neurofilament light chain KW - NfL KW - glial damage KW - cardiac magnetic resonance imaging KW - MRI KW - infarction size Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-288261 SN - 1422-0067 VL - 23 IS - 18 ER - TY - JOUR A1 - Weiss, Esther A1 - Schlegel, Jan A1 - Terpitz, Ulrich A1 - Weber, Michael A1 - Linde, Jörg A1 - Schmitt, Anna-Lena A1 - Hünniger, Kerstin A1 - Marischen, Lothar A1 - Gamon, Florian A1 - Bauer, Joachim A1 - Löffler, Claudia A1 - Kurzai, Oliver A1 - Morton, Charles Oliver A1 - Sauer, Markus A1 - Einsele, Hermann A1 - Loeffler, Juergen T1 - Reconstituting NK Cells After Allogeneic Stem Cell Transplantation Show Impaired Response to the Fungal Pathogen Aspergillus fumigatus JF - Frontiers in Immunology N2 - Delayed natural killer (NK) cell reconstitution after allogeneic stem cell transplantation (alloSCT) is associated with a higher risk of developing invasive aspergillosis. The interaction of NK cells with the human pathogen Aspergillus (A.) fumigatus is mediated by the fungal recognition receptor CD56, which is relocated to the fungal interface after contact. Blocking of CD56 signaling inhibits the fungal mediated chemokine secretion of MIP-1α, MIP-1β, and RANTES and reduces cell activation, indicating a functional role of CD56 in fungal recognition. We collected peripheral blood from recipients of an allograft at defined time points after alloSCT (day 60, 90, 120, 180). NK cells were isolated, directly challenged with live A. fumigatus germ tubes, and cell function was analyzed and compared to healthy age and gender-matched individuals. After alloSCT, NK cells displayed a higher percentage of CD56\(^{bright}\)CD16\(^{dim}\) cells throughout the time of blood collection. However, CD56 binding and relocalization to the fungal contact side were decreased. We were able to correlate this deficiency to the administration of corticosteroid therapy that further negatively influenced the secretion of MIP-1α, MIP-1β, and RANTES. As a consequence, the treatment of healthy NK cells ex vivo with corticosteroids abrogated chemokine secretion measured by multiplex immunoassay. Furthermore, we analyzed NK cells regarding their actin cytoskeleton by Structured Illumination Microscopy (SIM) and flow cytometry and demonstrate an actin dysfunction of NK cells shown by reduced F-actin content after fungal co-cultivation early after alloSCT. This dysfunction remains until 180 days post-alloSCT, concluding that further actin-dependent cellular processes may be negatively influenced after alloSCT. To investigate the molecular pathomechansism, we compared CD56 receptor mobility on the plasma membrane of healthy and alloSCT primary NK cells by single-molecule tracking. The results were very robust and reproducible between tested conditions which point to a different molecular mechanism and emphasize the importance of proper CD56 mobility. KW - natural killer cell KW - stem cell transplantation KW - corticosteroids KW - CCL3 KW - CCL4 KW - CCL5 Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212581 SN - 1664-3224 VL - 11 ER -