TY - JOUR A1 - Dech, Stefan A1 - Holzwarth, Stefanie A1 - Asam, Sarah A1 - Andresen, Thorsten A1 - Bachmann, Martin A1 - Boettcher, Martin A1 - Dietz, Andreas A1 - Eisfelder, Christina A1 - Frey, Corinne A1 - Gesell, Gerhard A1 - Gessner, Ursula A1 - Hirner, Andreas A1 - Hofmann, Matthias A1 - Kirches, Grit A1 - Klein, Doris A1 - Klein, Igor A1 - Kraus, Tanja A1 - Krause, Detmar A1 - Plank, Simon A1 - Popp, Thomas A1 - Reinermann, Sophie A1 - Reiners, Philipp A1 - Roessler, Sebastian A1 - Ruppert, Thomas A1 - Scherbachenko, Alexander A1 - Vignesh, Ranjitha A1 - Wolfmueller, Meinhard A1 - Zwenzner, Hendrik A1 - Kuenzer, Claudia T1 - Potential and challenges of harmonizing 40 years of AVHRR data: the TIMELINE experience JF - Remote Sensing N2 - Earth Observation satellite data allows for the monitoring of the surface of our planet at predefined intervals covering large areas. However, there is only one medium resolution sensor family in orbit that enables an observation time span of 40 and more years at a daily repeat interval. This is the AVHRR sensor family. If we want to investigate the long-term impacts of climate change on our environment, we can only do so based on data that remains available for several decades. If we then want to investigate processes with respect to climate change, we need very high temporal resolution enabling the generation of long-term time series and the derivation of related statistical parameters such as mean, variability, anomalies, and trends. The challenges to generating a well calibrated and harmonized 40-year-long time series based on AVHRR sensor data flown on 14 different platforms are enormous. However, only extremely thorough pre-processing and harmonization ensures that trends found in the data are real trends and not sensor-related (or other) artefacts. The generation of European-wide time series as a basis for the derivation of a multitude of parameters is therefore an extremely challenging task, the details of which are presented in this paper. KW - AVHRR KW - Earth Observation KW - harmonization KW - time series analysis KW - climate related trends KW - automatic processing KW - Europe KW - TIMELINE Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-246134 SN - 2072-4292 VL - 13 IS - 18 ER - TY - THES A1 - Klein, Philipp T1 - Kephalometrische Verlaufsuntersuchungen bei Kindern vor und nach operativer Versorgung einer isolierten Sagittalnahtsynostose T1 - Long term cephalometric follow up of children with operated isolated sagittal synostosis N2 - In dieser retrospektiven Arbeit wurde das Schädelwachstum von Patienten mit einer operierten nonsyndromalen prämaturen Sagittalnahtsynostose untersucht. Hierzu wurden die prä- und postoperativ angefertigten Röntgenaufnahmen von 37 Kindern, die zwischen 1995 und 2008 im Cranio-Fazialen Zentrum der Universitätsklinik Würzburg operiert wurden, vermessen und ausgewertet. Die Patienten wurden nach der gewählten Operationstechnik in zwei Gruppen unterteilt. Die erste Gruppe wurde mittels einer medianen Kraniektomie therapiert. Bei der zweiten Gruppe wurde die mediane Kraniektomie durch ein Kippen des Stirnsegmentes erweitert. Nach der statistischen Auswertung ergaben sich im postoperativen Verlauf signifikante Unterschiede zwischen beiden Operationstechniken. Es konnte gezeigt werden, dass ein adjuvantes Kippen der Stirn gegenüber einer alleinigen breiten medianen Kraniektomie zu keiner Verbesserung der Schädelausformung führt. Darüber hinaus ist zu vermuten, dass das Kippen des Stirnsegmentes die Wahrscheinlichkeit eines Rezidivs erhöht. N2 - In this retrospective work, cranial growth of patients with operated nonsyndromal prematurary sagittal suture synostosis was investigated. For this purpose, the pre- and postoperative radiographs of 37 children operated between 1995 and 2008 in the Cranio-Facial Center of the University Hospital Würzburg were measured and evaluated. The patients were divided into two groups according to the surgical technique chosen. The first group was treated with a median craniectomy. In the second group, median craniectomy was augmented by tilting the forehead segment. Statistical evaluation showed significant differences between the two surgical techniques during the postoperative period. It could be shown that an adjuvant tipping of the forehead compared to a sole wide median craniectomy does not lead to an improvement of the skull formation. In addition, it is likely that tilting the forehead segment increases the likelihood of recurrence. KW - Sagittalnahtsynostose KW - isoliert KW - Kephalometrie Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-188079 ER - TY - JOUR A1 - Klein, Philipp A1 - Johe, Patrick A1 - Wagner, Annika A1 - Jung, Sascha A1 - Kühlborn, Jonas A1 - Barthels, Fabian A1 - Tenzer, Stefan A1 - Distler, Ute A1 - Waigel, Waldemar A1 - Engels, Bernd A1 - Hellmich, Ute A. A1 - Opatz, Till A1 - Schirmeister, Tanja T1 - New cysteine protease inhibitors: electrophilic (het)arenes and unexpected prodrug identification for the Trypanosoma protease rhodesain JF - Molecules N2 - Electrophilic (het)arenes can undergo reactions with nucleophiles yielding π- or Meisenheimer (σ-) complexes or the products of the S\(_N\)Ar addition/elimination reactions. Such building blocks have only rarely been employed for the design of enzyme inhibitors. Herein, we demonstrate the combination of a peptidic recognition sequence with such electrophilic (het)arenes to generate highly active inhibitors of disease-relevant proteases. We further elucidate an unexpected mode of action for the trypanosomal protease rhodesain using NMR spectroscopy and mass spectrometry, enzyme kinetics and various types of simulations. After hydrolysis of an ester function in the recognition sequence of a weakly active prodrug inhibitor, the liberated carboxylic acid represents a highly potent inhibitor of rhodesain (K\(_i\) = 4.0 nM). The simulations indicate that, after the cleavage of the ester, the carboxylic acid leaves the active site and re-binds to the enzyme in an orientation that allows the formation of a very stable π-complex between the catalytic dyad (Cys-25/His-162) of rhodesain and the electrophilic aromatic moiety. The reversible inhibition mode results because the S\(_N\)Ar reaction, which is found in an alkaline solvent containing a low molecular weight thiol, is hindered within the enzyme due to the presence of the positively charged imidazolium ring of His-162. Comparisons between measured and calculated NMR shifts support this interpretation KW - cysteine protease KW - rhodesain KW - electrophilic (het)arene KW - nucleophilic aromatic substitution KW - Meisenheimer complex KW - π-complex KW - prodrug Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-203380 SN - 1420-3049 VL - 25 IS - 6 ER - TY - JOUR A1 - Klein, Philipp A1 - Barthels, Fabian A1 - Johe, Patrick A1 - Wagner, Annika A1 - Tenzer, Stefan A1 - Distler, Ute A1 - Le, Thien Anh A1 - Schmid, Paul A1 - Engel, Volker A1 - Engels, Bernd A1 - Hellmich, Ute A. A1 - Opatz, Till A1 - Schirmeister, Tanja T1 - Naphthoquinones as covalent reversible inhibitors of cysteine proteases — studies on inhibition mechanism and kinetics JF - Molecules N2 - The facile synthesis and detailed investigation of a class of highly potent protease inhibitors based on 1,4-naphthoquinones with a dipeptidic recognition motif (HN-l-Phe-l-Leu-OR) in the 2-position and an electron-withdrawing group (EWG) in the 3-position is presented. One of the compound representatives, namely the acid with EWG = CN and with R = H proved to be a highly potent rhodesain inhibitor with nanomolar affinity. The respective benzyl ester (R = Bn) was found to be hydrolyzed by the target enzyme itself yielding the free acid. Detailed kinetic and mass spectrometry studies revealed a reversible covalent binding mode. Theoretical calculations with different density functionals (DFT) as well as wavefunction-based approaches were performed to elucidate the mode of action. KW - protease KW - rhodesain KW - covalent reversible inhibition KW - 1,4-naphthoquinone KW - nucleophilic addition KW - prodrug Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-203791 SN - 1420-3049 VL - 25 IS - 9 ER - TY - JOUR A1 - Summa, Michela A1 - Klein, Martin A1 - Schmidt, Philipp T1 - Introduction: Double Intentionality JF - Topoi N2 - No abstract available. KW - double intentionality KW - intentional directions KW - experiences Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-269865 SN - 1572-8749 VL - 41 IS - 1 ER - TY - JOUR A1 - Dietschreit, Johannes C. B. A1 - Wagner, Annika A1 - Le, T. Anh A1 - Klein, Philipp A1 - Schindelin, Hermann A1 - Opatz, Till A1 - Engels, Bernd A1 - Hellmich, Ute A. A1 - Ochsenfeld, Christian T1 - Predicting \(^{19}\)F NMR Chemical Shifts: A Combined Computational and Experimental Study of a Trypanosomal Oxidoreductase–Inhibitor Complex JF - Angewandte Chemie International Edition N2 - The absence of fluorine from most biomolecules renders it an excellent probe for NMR spectroscopy to monitor inhibitor–protein interactions. However, predicting the binding mode of a fluorinated ligand from a chemical shift (or vice versa) has been challenging due to the high electron density of the fluorine atom. Nonetheless, reliable \(^{19}\)F chemical‐shift predictions to deduce ligand‐binding modes hold great potential for in silico drug design. Herein, we present a systematic QM/MM study to predict the \(^{19}\)F NMR chemical shifts of a covalently bound fluorinated inhibitor to the essential oxidoreductase tryparedoxin (Tpx) from African trypanosomes, the causative agent of African sleeping sickness. We include many protein–inhibitor conformations as well as monomeric and dimeric inhibitor–protein complexes, thus rendering it the largest computational study on chemical shifts of \(^{19}\)F nuclei in a biological context to date. Our predicted shifts agree well with those obtained experimentally and pave the way for future work in this area. KW - African sleeping sickness KW - covalent inhibitors KW - NMR spectroscopy KW - quantum chemistry KW - structural biology Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-214879 VL - 59 IS - 31 SP - 12669 EP - 12673 ER - TY - JOUR A1 - Krajka, Victor A1 - Naujock, Maximilian A1 - Pauly, Martje G. A1 - Stengel, Felix A1 - Meier, Britta A1 - Stanslowsky, Nancy A1 - Klein, Christine A1 - Seibler, Philip A1 - Wegner, Florian A1 - Capetian, Philipp T1 - Ventral Telencephalic Patterning Protocols for Induced Pluripotent Stem Cells JF - Frontiers in Cell and Developmental Biology N2 - The differentiation of human induced pluripotent stem cells (hiPSCs) into specific cell types for disease modeling and restorative therapies is a key research agenda and offers the possibility to obtain patient-specific cells of interest for a wide range of diseases. Basal forebrain cholinergic neurons (BFCNs) play a particular role in the pathophysiology of Alzheimer’s dementia and isolated dystonias. In this work, various directed differentiation protocols based on monolayer neural induction were tested for their effectiveness in promoting a ventral telencephalic phenotype and generating BFCN. Ventralizing factors [i.e., purmorphamine and Sonic hedgehog (SHH)] were applied at different time points, time intervals, and concentrations. In addition, caudal identity was prevented by the use of a small molecule XAV-939 that inhibits the Wnt-pathway. After patterning, gene expression profiles were analyzed by quantitative PCR (qPCR). Rostro-ventral patterning is most effective when initiated simultaneously with neural induction. The most promising combination of patterning factors was 0.5 μM of purmorphamine and 1 μM of XAV-939, which induces the highest expression of transcription factors specific for the medial ganglionic eminence, the source of GABAergic inter- and cholinergic neurons in the telencephalon. Upon maturation of cells, the immune phenotype, as well as electrophysiological properties were investigated showing the presence of marker proteins specific for BFCN (choline acetyltransferase, ISL1, p75, and NKX2.1) and GABAergic neurons. Moreover, a considerable fraction of measured cells displayed mature electrophysiological properties. Synaptic boutons containing the vesicular acetylcholine transporter (VACHT) could be observed in the vicinity of the cells. This work will help to generate basal forebrain interneurons from hiPSCs, providing a promising platform for modeling neurological diseases, such as Alzheimer’s disease or Dystonia. KW - induced pluripotent stem cells KW - medial ganglionic eminence KW - Sonic hedgehog KW - XAV-939 KW - purmorphamine KW - basal forebrain cholinergic neurons KW - GABAergic neurons KW - electrophysiology Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244607 SN - 2296-634X VL - 9 ER - TY - JOUR A1 - Stengel, Felix A1 - Vulinovic, Franca A1 - Meier, Britta A1 - Grütz, Karen A1 - Klein, Christine A1 - Capetian, Philipp T1 - Impaired differentiation of human induced neural stem cells by TOR1A overexpression JF - Molecular Biology Reports N2 - DYT-TOR1A is the most common inherited dystonia caused by a three nucleotide (GAG) deletion (dE) in the TOR1A gene. Death early after birth and cortical anomalies of the full knockout in rodents underscore its developmental importance. We therefore explored the timed effects of TOR1A-wt and TOR1A-dE during differentiation in a human neural in vitro model. We used lentiviral tet-ON expression of TOR1A-wt and -dE in induced neural stem cells derived from healthy donors. Overexpression was induced during proliferation of neural precursors, during differentiation and after differentiation into mature neurons. Overexpression of both wildtype and mutated protein had no effect on the viability and cell number of neural precursors as well as mature neurons when initiated before or after differentiation. However, if induced during differentiation, overexpression of TOR1A-wt and -dE led to a pronounced reduction of mature neurons in a dose dependent manner. Our data underscores the importance of physiological expression levels of TOR1A as crucial for proper neuronal differentiation. We did not find evidence for a specific impact of the mutated TOR1A on neuronal maturation. KW - dystonia KW - DYT1 KW - torsinA KW - TOR1A KW - neuronal stem cells KW - neuronal differentiation KW - inducible expression Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241177 UR - https://doi.org/10.1007/s11033-020-05390-x VL - 47 ER -