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    Precision 3D‐printed cell scaffolds mimicking native tissue composition and mechanics
    (2020) Erben, Amelie; Hörning, Marcel; Hartmann, Bastian; Becke, Tanja; Eisler, Stephan A.; Southan, Alexander; Cranz, Séverine; Hayden, Oliver; Kneidinger, Nikolaus; Königshoff, Melanie; Lindner, Michael; Tovar, Günter E. M.; Burgstaller, Gerald; Clausen‐Schaumann, Hauke; Sudhop, Stefanie; Heymann, Michael
    Cellular dynamics are modeled by the 3D architecture and mechanics of the extracellular matrix (ECM) and vice versa. These bidirectional cell‐ECM interactions are the basis for all vital tissues, many of which have been investigated in 2D environments over the last decades. Experimental approaches to mimic in vivo cell niches in 3D with the highest biological conformity and resolution can enable new insights into these cell‐ECM interactions including proliferation, differentiation, migration, and invasion assays. Here, two‐photon stereolithography is adopted to print up to mm‐sized high‐precision 3D cell scaffolds at micrometer resolution with defined mechanical properties from protein‐based resins, such as bovine serum albumin or gelatin methacryloyl. By modifying the manufacturing process including two‐pass printing or post‐print crosslinking, high precision scaffolds with varying Young's moduli ranging from 7‐300 kPa are printed and quantified through atomic force microscopy. The impact of varying scaffold topographies on the dynamics of colonizing cells is observed using mouse myoblast cells and a 3D‐lung microtissue replica colonized with primary human lung fibroblast. This approach will allow for a systematic investigation of single‐cell and tissue dynamics in response to defined mechanical and bio‐molecular cues and is ultimately scalable to full organs.
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    Messungen im Bereich eines Windparks mit Fokus auf tief- und niederfrequente Schallemissionen und -immissionen
    (2022) Blumendeller, Esther; Gaßner, Laura; Müller, Florian; Wigger, Maayen; Berlinger, Philipp; Cheng, Po Wen
    Die Nutzung von Windenergie wird einen entscheidenden Anteil am erneuerbaren Energiemix der Zukunft haben. Während der Stromgewinnung geben Windenergieanlagen (WEA) Schall und Erschütterungen (seismische Wellen) in die Umgebung ab, vor allem im tieffrequenten Bereich. Im Zuge des interdisziplinären Verbundprojektes Inter-Wind werden akustische Messungen zur Unterstützung psychologischer Fragebögen, kombiniert mit seismischen und meteorologischen Messungen an Windparks auf der Schwäbischen Alb durchgeführt. Ziel des Projektes ist es, die Gründe für Belästigung der Anwohner in Zusammenhang mit den Immissionen der WEA zu verstehen. Hierbei liegt der Fokus auf dem tieffrequenten (20-200 Hz) und niederfrequenten (1-20 Hz) Bereich. Akustische und seismische Messungen wurden an einem Windpark auf der Schwäbischen Alb, mit drei WEA des Typs GE 2.75-120 durchgeführt. Parallel dazu konnten Anwohner Belästigungszeiträume über eine Geräuschmelde-App dokumentieren. In diesem Beitrag wird die Umsetzung einer interdisziplinären Messkampagne im Bereich des Tegelberg Windparks und eines Wohngebäudes in Tallage in ca. 1 km Entfernung zum Windpark beschrieben. Schließlich werden erste Ergebnisse der akustischen Messungen und interdisziplinären Untersuchung vorgestellt und diskutiert.
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    Magnetic Moment Tensor Potentials for collinear spin-polarized materials reproduce different magnetic states of bcc Fe
    (2022) Novikov, Ivan; Grabowski, Blazej; Körmann, Fritz; Shapeev, Alexander
    We present the magnetic Moment Tensor Potentials (mMTPs), a class of machine-learning interatomic potentials, accurately reproducing both vibrational and magnetic degrees of freedom as provided, e.g., from first-principles calculations. The accuracy is achieved by a two-step minimization scheme that coarse-grains the atomic and the spin space. The performance of the mMTPs is demonstrated for the prototype magnetic system bcc iron, with applications to phonon calculations for different magnetic states, and molecular-dynamics simulations with fluctuating magnetic moments.
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    Eclectic characterisation of chemically modified cell-derived matrices obtained by metabolic glycoengineering and re-assessment of commonly used methods
    (2020) Keller, Silke; Liedek, Anke; Shendi, Dalia; Bach, Monika; Tovar, Günter E. M.; Kluger, Petra J.; Southan, Alexander
    Azide-bearing cell-derived extracellular matrices (“clickECMs”) have emerged as a highly exciting new class of biomaterials. They conserve substantial characteristics of the natural extracellular matrix (ECM) and offer simultaneously small abiotic functional groups that enable bioorthogonal bioconjugation reactions. Despite their attractiveness, investigation of their biomolecular composition is very challenging due to the insoluble and highly complex nature of cell-derived matrices (CDMs). Yet, thorough qualitative and quantitative analysis of the overall material composition, organisation, localisation, and distribution of typical ECM-specific biomolecules is essential for consistent advancement of CDMs and the understanding of the prospective functions of the developed biomaterial. In this study, we evaluated frequently used methods for the analysis of complex CDMs. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and (immune)histochemical staining methods in combination with several microscopic techniques were found to be highly eligible. Commercially available colorimetric protein assays turned out to deliver inaccurate information on CDMs. In contrast, we determined the nitrogen content of CDMs by elementary analysis and converted it into total protein content using conversion factors which were calculated from matching amino acid compositions. The amount of insoluble collagens was assessed based on the hydroxyproline content. The Sircol™ assay was identified as a suitable method to quantify soluble collagens while the Blyscan™ assay was found to be well-suited for the quantification of sulphated glycosaminoglycans (sGAGs). Eventually, we propose a series of suitable methods to reliably characterise the biomolecular composition of fibroblast-derived clickECM.
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    Comparative sensitivity analysis of muscle activation dynamics
    (2015) Rockenfeller, Robert; Günther, Michael; Schmitt, Syn; Götz, Thomas
    We mathematically compared two models of mammalian striated muscle activation dynamics proposed by Hatze and Zajac. Both models are representative for a broad variety of biomechanical models formulated as ordinary differential equations (ODEs). These models incorporate parameters that directly represent known physiological properties. Other parameters have been introduced to reproduce empirical observations. We used sensitivity analysis to investigate the influence of model parameters on the ODE solutions. In addition, we expanded an existing approach to treating initial conditions as parameters and to calculating second- order sensitivities. Furthermore, we used a global sensitivity analysis approach to include finite ranges of parameter values. Hence, a theoretician striving for model reduction could use the method for identifying particularly low sensitivities to detect superfluous parameters. An experimenter could use it for identifying particularly high sensitivities to improve parameter estimation. Hatze’s nonlinear model incorporates some parameters to which activation dynamics is clearly more sensitive than to any parameter in Zajac’s linear model. Other than Zajac’s model, Hatze’s model can, however, reproduce measured shifts in optimal muscle length with varied muscle activity. Accordingly we extracted a specific parameter set for Hatze’s model that combines best with a particular muscle force-length relation.
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    Differentiation of physical and chemical cross-linking in gelatin methacryloyl hydrogels
    (2021) Rebers, Lisa; Reichsöllner, Raffael; Regett, Sophia; Tovar, Günter E. M.; Borchers, Kirsten; Baudis, Stefan; Southan, Alexander
    Gelatin methacryloyl (GM) hydrogels have been investigated for almost 20 years, especially for biomedical applications. Recently, strengthening effects of a sequential cross-linking procedure, whereby GM hydrogel precursor solutions are cooled before chemical cross-linking, were reported. It was hypothesized that physical and enhanced chemical cross-linking of the GM hydrogels contribute to the observed strengthening effects. However, a detailed investigation is missing so far. In this contribution, we aimed to reveal the impact of physical and chemical cross-linking on strengthening of sequentially cross-linked GM and gelatin methacryloyl acetyl (GMA) hydrogels. We investigated physical and chemical cross-linking of three different GM(A) derivatives (GM10, GM2A8 and GM2), which provided systematically varied ratios of side-group modifications. GM10 contained the highest methacryloylation degree (DM), reducing its ability to cross-link physically. GM2 had the lowest DM and showed physical cross-linking. The total modification degree, determining the physical cross-linking ability, of GM2A8 was comparable to that of GM10, but the chemical cross-linking ability was comparable to GM2. At first, we measured the double bond conversion (DBC) kinetics during chemical GM(A) cross-linking quantitatively in real-time via near infrared spectroscopy-photorheology and showed that the DBC decreased due to sequential cross-linking. Furthermore, results of circular dichroism spectroscopy and differential scanning calorimetry indicated gelation and conformation changes, which increased storage moduli of all GM(A) hydrogels due to sequential cross-linking. The data suggested that the total cross-link density determines hydrogel stiffness, regardless of the physical or chemical nature of the cross-links.
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    Operational scenarios optimization for resupply of crew and cargo of an International gateway Station located near the Earth-Moon-Lagrangian point-2
    (2016) Lizy-Destrez, Stéphanie; Messerschmid, Ernst (Prof. Dr.)
    In the context of future human space exploration missions in the solar system (with an horizon of 2025) and according to the roadmap proposed by ISECG (International Space Exploration Coordination Group) [1], a new step could be to maintain as an outpost, at one of the libration points of the Earth-Moon system, a space station. This would ease access to far destinations as Moon, Mars and asteroids and would allow testing some innovative technologies, before employing them for far distant human missions. One of the main challenges will be to maintain permanently, and ensure on board crew health thanks to an autonomous space medical center docked to the proposed space station, as a Space haven. Then the main problem to solve is to manage the station servitude, during deployment (modules integration) and operational phase. Challenges lie, on a global point of view, in the design of the operational scenarios and, on a local point of view, in trajectories selection, so as to minimize velocity increments (energy consumption) and transportation duration (crew safety). Which recommendations could be found out as far as trajectories optimization is concerned, that would fulfill energy consumption, transportation duration and safety criterion? What would technological hurdles be to rise for the building of such Space haven? What would be performances to aim at for critical sub-systems? Expected results of this study could point out research and development perspectives for human spaceflight missions and above all, in transportation field for long lasting missions. Thus, the thesis project, presented here, aims starting from global system life-cycle decomposition, to identify by phase operational scenario and optimize resupply vehicle mission.
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    The PedS2/PedR2 two-component system is crucial for the rare earth element switch in Pseudomonas putida KT2440
    (2018) Wehrmann, Matthias; Berthelot, Charlotte; Billard, Patrick; Klebensberger, Janosch
    ABSTRACT In Pseudomonas putida KT2440, two pyrroloquinoline quinone-dependent ethanol dehydrogenases (PQQ-EDHs) are responsible for the periplasmic oxidation of a broad variety of volatile organic compounds (VOCs). Depending on the availability of rare earth elements (REEs) of the lanthanide series (Ln3+), we have recently reported that the transcription of the genes encoding the Ca2+-utilizing enzyme PedE and the Ln3+-utilizing enzyme PedH are inversely regulated. With adaptive evolution experiments, site-specific mutations, transcriptional reporter fusions, and complementation approaches, we now demonstrate that the PedS2/PedR2 (PP_2671/PP_2672) two-component system (TCS) plays a central role in the observed REE-mediated switch of PQQ-EDHs in P. putida. We provide evidence that in the absence of lanthanum (La3+), the sensor histidine kinase PedS2 phosphorylates its cognate LuxR-type response regulator PedR2, which in turn not only activates pedE gene transcription but is also involved in repression of pedH. Our data further suggest that the presence of La3+ lowers kinase activity of PedS2, either by the direct binding of the metal ions to the periplasmic region of PedS2 or by an uncharacterized indirect interaction, leading to reduced levels of phosphorylated PedR2. Consequently, the decreasing pedE expression and concomitant alleviation of pedH repression causes - in conjunction with the transcriptional activation of the pedH gene by a yet unknown regulatory module - the Ln3+-dependent transition from PedE- to PedH-catalyzed oxidation of alcoholic VOCs. IMPORTANCE The function of lanthanides for methanotrophic and methylotrophic bacteria is gaining increasing attention, while knowledge about the role of rare earth elements (REEs) in nonmethylotrophic bacteria is still limited. The present study investigates the recently described differential expression of the two PQQ-EDHs of P. putida in response to lanthanides. We demonstrate that a specific TCS is crucial for their inverse regulation and provide evidence for a dual regulatory function of the LuxR-type response regulator involved. Thus, our study represents the first detailed characterization of the molecular mechanism underlying the REE switch of PQQ-EDHs in a nonmethylotrophic bacterium and stimulates subsequent investigations for the identification of additional genes or phenotypic traits that might be coregulated during REE-dependent niche adaptation.
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    Crop water productivity mapping and benchmarking using remote sensing and Google Earth Engine cloud computing
    (2022) Ghorbanpour, Ali Karbalaye; Kisekka, Isaya; Afshar, Abbas; Hessels, Tim; Taraghi, Mahdi; Hessari, Behzad; Tourian, Mohammad J.; Duan, Zheng
    Scarce water resources present a major hindrance to ensuring food security. Crop water productivity (WP), embraced as one of the Sustainable Development Goals (SDGs), is playing an integral role in the performance-based evaluation of agricultural systems and securing sustainable food production. This study aims at developing a cloud-based model within the Google Earth Engine (GEE) based on Landsat -7 and -8 satellite imagery to facilitate WP mapping at regional scales (30-m resolution) and analyzing the state of the water use efficiency and productivity of the agricultural sector as a means of benchmarking its WP and defining local gaps and targets at spatiotemporal scales. The model was tested in three major agricultural districts in the Lake Urmia Basin (LUB) with respect to five crop types, including irrigated wheat, rainfed wheat, apples, grapes, alfalfa, and sugar beets as the major grown crops. The actual evapotranspiration (ET) was estimated using geeSEBAL based on the Surface Energy Balance Algorithm for Land (SEBAL) methodology, while for crop yield estimations Monteith’s Light Use Efficiency model (LUE) was employed. The results indicate that the WP in the LUB is below its optimum targets, revealing that there is a significant degree of work necessary to ameliorate the WP in the LUB. The WP varies between 0.49-0.55 (kg/m3) for irrigated wheat, 0.27-0.34 for rainfed wheat, 1.7-2.2 for apples, 1.2-1.7 for grapes, 5.5-6.2 for sugar beets, and 0.67-1.08 for alfalfa, which could be potentially increased up to 80%, 150%, 76%, 83%, 55%, and 48%, respectively. The spatial variation of the WP and crop yield makes it feasible to detect the areas with the best and poorest on-farm practices, thereby facilitating the better targeting of resources to bridge the WP gap through water management practices. This study provides important insights into the status and potential of WP with possible worldwide applications at both farm and government levels for policymakers, practitioners, and growers to adopt effective policy guidelines and improve on-farm practices.
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    Valorization of pyrolysis water: a biorefinery side stream, for 1,2‑propanediol production with engineered Corynebacterium glutamicum
    (2017) Lange, Julian; Müller, Felix; Bernecker, Kerstin; Dahmen, Nicolaus; Takors, Ralf; Blombach, Bastian