TY - JOUR A1 - Haeger, Gerrit A1 - Grankin, Alina A1 - Wagner, Michaela T1 - Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology JF - Applied Research N2 - Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems. KW - aspergillus KW - CRISPR/Cas9 KW - filamentous fungi KW - genome engineering Y1 - 2023 U6 - http://dx.doi.org/10.1002/appl.202200106 SN - 2702-4288 IS - Early View SP - 1 EP - 15 PB - Wiley-VCH ER - TY - JOUR A1 - Bruksle, Ieva A1 - Chwallek, Constanze A1 - Krastina, Anzelika T1 - Strengthening sustainability in entrepreneurship education - implications for shifting entrepreneurial thinking towards sustainability at universities JF - ACTA PROSPERITATIS N2 - By developing innovative solutions to social and environmental problems, sustainable ventures carry greatpotential. Entrepreneurship which focuses especially on new venture creation can be developed through education anduniversities, in particular, are called upon to provide an impetus for social change. But social innovations are associatedwith certain hurdles, which are related to the multi-dimensionality, i.e. the tension between creating social,environmental and economic value and dealing with a multiplicity of stakeholders. The already complex field ofentrepreneurship education has to face these challenges. This paper, therefore, aims to identify starting points for theintegration of sustainability into entrepreneurship education. To pursue this goal experiences from three differentproject initiatives between the partner universities: Lapland University of Applied Sciences, FH Aachen University ofApplied Sciences and Turiba University are reflected and findings are systematically condensed into recommendationsfor education on sustainable entrepreneurship. KW - climate change KW - entrepreneurship education KW - Finland KW - Germany KW - Latvia Y1 - 2023 U6 - http://dx.doi.org/10.37804/1691-6077-2023-14-37-48 SN - 1691-6077 VL - 14 IS - 1 SP - 37 EP - 48 PB - Sciendo ER - TY - JOUR A1 - Savitskaya, Irina A1 - Zhantlessova, Sirina A1 - Kistaubayeva, Aida A1 - Ignatova, Ludmila A1 - Shokatayeva, Dina A1 - Sinyavsky, Yuriy A1 - Kushugulova, Almagul A1 - Digel, Ilya T1 - Prebiotic cellulose–pullulan matrix as a “vehicle” for probiotic biofilm delivery to the host large intestine JF - Polymers N2 - This study describes the development of a new combined polysaccharide-matrix-based technology for the immobilization of Lactobacillus rhamnosus GG (LGG) bacteria in biofilm form. The new composition allows for delivering the bacteria to the digestive tract in a manner that improves their robustness compared with planktonic cells and released biofilm cells. Granules consisting of a polysaccharide matrix with probiotic biofilms (PMPB) with high cell density (>9 log CFU/g) were obtained by immobilization in the optimized nutrient medium. Successful probiotic loading was confirmed by fluorescence microscopy and scanning electron microscopy. The developed prebiotic polysaccharide matrix significantly enhanced LGG viability under acidic (pH 2.0) and bile salt (0.3%) stress conditions. Enzymatic extract of feces, mimicking colon fluid in terms of cellulase activity, was used to evaluate the intestinal release of probiotics. PMPB granules showed the ability to gradually release a large number of viable LGG cells in the model colon fluid. In vivo, the oral administration of PMPB granules in rats resulted in the successful release of probiotics in the colon environment. The biofilm-forming incubation method of immobilization on a complex polysaccharide matrix tested in this study has shown high efficacy and promising potential for the development of innovative biotechnologies. KW - immobilization KW - prebiotic KW - bacterial cellulose KW - biofilms KW - Lactobacillus rhamnosus GG Y1 - 2023 U6 - http://dx.doi.org/10.3390/polym16010030 N1 - This article belongs to the Section "Polymer Composites and Nanocomposites" IS - 16(1) PB - MDPI CY - Basel ER - TY - CHAP A1 - Maurer, Florian A1 - Miskiw, Kim K. A1 - Acosta, Rebeca Ramirez A1 - Harder, Nick A1 - Sander, Volker A1 - Lehnhoff, Sebastian ED - Jorgensen, Bo Norregaard ED - Pereira da Silva, Luiz Carlos ED - Ma, Zheng T1 - Market abstraction of energy markets and policies - application in an agent-based modeling toolbox T2 - EI.A 2023: Energy Informatics N2 - In light of emerging challenges in energy systems, markets are prone to changing dynamics and market design. Simulation models are commonly used to understand the changing dynamics of future electricity markets. However, existing market models were often created with specific use cases in mind, which limits their flexibility and usability. This can impose challenges for using a single model to compare different market designs. This paper introduces a new method of defining market designs for energy market simulations. The proposed concept makes it easy to incorporate different market designs into electricity market models by using relevant parameters derived from analyzing existing simulation tools, morphological categorization and ontologies. These parameters are then used to derive a market abstraction and integrate it into an agent-based simulation framework, allowing for a unified analysis of diverse market designs. Furthermore, we showcase the usability of integrating new types of long-term contracts and over-the-counter trading. To validate this approach, two case studies are demonstrated: a pay-as-clear market and a pay-as-bid long-term market. These examples demonstrate the capabilities of the proposed framework. KW - Energy market design KW - Agent-based simulation KW - Market modeling Y1 - 2023 SN - 978-3-031-48651-7 (Print) SN - 978-3-031-48652-4 (eBook) U6 - http://dx.doi.org/10.1007/978-3-031-48652-4_10 N1 - Energy Informatics Academy Conference, 6-8 December 23, Campinas, Brazil. N1 - Part of the Lecture Notes in Computer Science book series (LNCS,volume 14468). SP - 139 EP - 157 PB - Springer CY - Cham ER - TY - JOUR A1 - Vögele, Stefan A1 - Josyabhatla, Vishnu Teja A1 - Ball, Christopher A1 - Rhoden, Imke A1 - Grajewski, Matthias A1 - Rübbelke, Dirk A1 - Kuckshinrichs, Wilhelm T1 - Robust assessment of energy scenarios from stakeholders' perspectives JF - Energy N2 - Using scenarios is vital in identifying and specifying measures for successfully transforming the energy system. Such transformations can be particularly challenging and require the support of a broader set of stakeholders. Otherwise, there will be opposition in the form of reluctance to adopt the necessary technologies. Usually, processes for considering stakeholders' perspectives are very time-consuming and costly. In particular, there are uncertainties about how to deal with modifications in the scenarios. In principle, new consulting processes will be required. In our study, we show how multi-criteria decision analysis can be used to analyze stakeholders' attitudes toward transition paths. Since stakeholders differ regarding their preferences and time horizons, we employ a multi-criteria decision analysis approach to identify which stakeholders will support or oppose a transition path. We provide a flexible template for analyzing stakeholder preferences toward transition paths. This flexibility comes from the fact that our multi-criteria decision aid-based approach does not involve intensive empirical work with stakeholders. Instead, it involves subjecting assumptions to robustness analysis, which can help identify options to influence stakeholders' attitudes toward transitions. Y1 - 2023 U6 - http://dx.doi.org/10.1016/j.energy.2023.128326 SN - 1873-6785 (Online) SN - 0360-5442 (Print) IS - In Press, Article 128326 PB - Elsevier CY - Amsterdam ER - TY - CHAP A1 - Goh, Kheng Lim A1 - Topçu, Murat A1 - Madabhushi, Gopal S. P. A1 - Staat, Manfred ED - Maia, Fatima Raquel Azevedo ED - Miguel Oliveira, J. ED - Reis, Rui L. T1 - Collagen fibril reinforcement in connective tissue extracellular matrices T2 - Handbook of the extracellular matrix N2 - The connective tissues such as tendons contain an extracellular matrix (ECM) comprising collagen fibrils scattered within the ground substance. These fibrils are instrumental in lending mechanical stability to tissues. Unfortunately, our understanding of how collagen fibrils reinforce the ECM remains limited, with no direct experimental evidence substantiating current theories. Earlier theoretical studies on collagen fibril reinforcement in the ECM have relied predominantly on the assumption of uniform cylindrical fibers, which is inadequate for modelling collagen fibrils, which possessed tapered ends. Recently, Topçu and colleagues published a paper in the International Journal of Solids and Structures, presenting a generalized shear-lag theory for the transfer of elastic stress between the matrix and fibers with tapered ends. This paper is a positive step towards comprehending the mechanics of the ECM and makes a valuable contribution to formulating a complete theory of collagen fibril reinforcement in the ECM. KW - Connective tissues KW - Extracellular matrix (ECM) KW - Collagen fibrils KW - Mechanical stability KW - Tapered ends Y1 - 2023 SN - 978-3-030-92090-6 (Print) SN - 978-3-030-92090-6 (Online) U6 - http://dx.doi.org/10.1007/978-3-030-92090-6_6-1 SP - 1 EP - 20 PB - Springer Nature CY - Cham ER - TY - JOUR A1 - Schulze, Sven A1 - Feyerl, Günter A1 - Pischinger, Stefan T1 - Advanced ECMS for hybrid electric heavy-duty trucks with predictive battery discharge and adaptive operating strategy under real driving conditions JF - Energies N2 - To fulfil the CO2 emission reduction targets of the European Union (EU), heavy-duty (HD) trucks need to operate 15% more efficiently by 2025 and 30% by 2030. Their electrification is necessary as conventional HD trucks are already optimized for the long-haul application. The resulting hybrid electric vehicle (HEV) truck gains most of the fuel saving potential by the recuperation of potential energy and its consecutive utilization. The key to utilizing the full potential of HEV-HD trucks is to maximize the amount of recuperated energy and ensure its intelligent usage while keeping the operating point of the internal combustion engine as efficient as possible. To achieve this goal, an intelligent energy management strategy (EMS) based on ECMS is developed for a parallel HEV-HD truck which uses predictive discharge of the battery and adaptive operating strategy regarding the height profile and the vehicle mass. The presented EMS can reproduce the global optimal operating strategy over long phases and lead to a fuel saving potential of up to 2% compared with a heuristic strategy. Furthermore, the fuel saving potential is correlated with the investigated boundary conditions to deepen the understanding of the impact of intelligent EMS for HEV-HD trucks. KW - Energy management strategies KW - ECMS KW - CO2 emission reduction targets KW - Driving cycle recognition KW - Predictive battery discharge Y1 - 2023 U6 - http://dx.doi.org/10.3390/en16135171 SN - 1996-1073 N1 - The article belongs to the Special Issue "Energy Management Strategies of Electrified Vehicles toward the Real-World Driving". VL - 16 IS - 13 PB - MDPI CY - Basel ER - TY - CHAP A1 - Büsgen, André A1 - Klöser, Lars A1 - Kohl, Philipp A1 - Schmidts, Oliver A1 - Kraft, Bodo A1 - Zündorf, Albert ED - Cuzzocrea, Alfredo ED - Gusikhin, Oleg ED - Hammoudi, Slimane ED - Quix, Christoph T1 - From cracked accounts to fake IDs: user profiling on German telegram black market channels T2 - Data Management Technologies and Applications N2 - Messenger apps like WhatsApp and Telegram are frequently used for everyday communication, but they can also be utilized as a platform for illegal activity. Telegram allows public groups with up to 200.000 participants. Criminals use these public groups for trading illegal commodities and services, which becomes a concern for law enforcement agencies, who manually monitor suspicious activity in these chat rooms. This research demonstrates how natural language processing (NLP) can assist in analyzing these chat rooms, providing an explorative overview of the domain and facilitating purposeful analyses of user behavior. We provide a publicly available corpus of annotated text messages with entities and relations from four self-proclaimed black market chat rooms. Our pipeline approach aggregates the extracted product attributes from user messages to profiles and uses these with their sold products as features for clustering. The extracted structured information is the foundation for further data exploration, such as identifying the top vendors or fine-granular price analyses. Our evaluation shows that pretrained word vectors perform better for unsupervised clustering than state-of-the-art transformer models, while the latter is still superior for sequence labeling. KW - Clustering KW - Natural language processing KW - Information extraction KW - Profile extraction KW - Text mining Y1 - 2023 SN - 978-3-031-37889-8 (Print) SN - 978-3-031-37890-4 (Online) U6 - http://dx.doi.org/10.1007/978-3-031-37890-4_9 N1 - 10th International Conference, DATA 2021, Virtual Event, July 6–8, 2021, and 11th International Conference, DATA 2022, Lisbon, Portugal, July 11-13, 2022 SP - 176 EP - 202 PB - Springer CY - Cham ER - TY - JOUR A1 - Heieis, Jule A1 - Böcker, Jonas A1 - D'Angelo, Olfa A1 - Mittag, Uwe A1 - Albracht, Kirsten A1 - Schönau, Eckhard A1 - Meyer, Andreas A1 - Voigtmann, Thomas A1 - Rittweger, Jörn T1 - Curvature of gastrocnemius muscle fascicles as function of muscle–tendon complex length and contraction in humans JF - Physiological Reports N2 - It has been shown that muscle fascicle curvature increases with increasing contraction level and decreasing muscle–tendon complex length. The analyses were done with limited examination windows concerning contraction level, muscle–tendon complex length, and/or intramuscular position of ultrasound imaging. With this study we aimed to investigate the correlation between fascicle arching and contraction, muscle–tendon complex length and their associated architectural parameters in gastrocnemius muscles to develop hypotheses concerning the fundamental mechanism of fascicle curving. Twelve participants were tested in five different positions (90°/105°*, 90°/90°*, 135°/90°*, 170°/90°*, and 170°/75°*; *knee/ankle angle). They performed isometric contractions at four different contraction levels (5%, 25%, 50%, and 75% of maximum voluntary contraction) in each position. Panoramic ultrasound images of gastrocnemius muscles were collected at rest and during constant contraction. Aponeuroses and fascicles were tracked in all ultrasound images and the parameters fascicle curvature, muscle–tendon complex strain, contraction level, pennation angle, fascicle length, fascicle strain, intramuscular position, sex and age group were analyzed by linear mixed effect models. Mean fascicle curvature of the medial gastrocnemius increased with contraction level (+5 m−1 from 0% to 100%; p = 0.006). Muscle–tendon complex length had no significant impact on mean fascicle curvature. Mean pennation angle (2.2 m−1 per 10°; p < 0.001), inverse mean fascicle length (20 m−1 per cm−1; p = 0.003), and mean fascicle strain (−0.07 m−1 per +10%; p = 0.004) correlated with mean fascicle curvature. Evidence has also been found for intermuscular, intramuscular, and sex-specific intramuscular differences of fascicle curving. Pennation angle and the inverse fascicle length show the highest predictive capacities for fascicle curving. Due to the strong correlations between pennation angle and fascicle curvature and the intramuscular pattern of curving we suggest for future studies to examine correlations between fascicle curvature and intramuscular fluid pressure. KW - biomechanics KW - connective tissue KW - physiology KW - ultrasound Y1 - 2023 U6 - http://dx.doi.org/10.14814/phy2.15739 SN - 2051-817X VL - 11 IS - 11 SP - e15739, Seite 1-11 PB - Wiley ER - TY - JOUR A1 - Kuchler, Timon A1 - Günthner, Roman A1 - Ribeiro, Andrea A1 - Hausinger, Renate A1 - Streese, Lukas A1 - Wöhnl, Anna A1 - Kesseler, Veronika A1 - Negele, Johanna A1 - Assali, Tarek A1 - Carbajo-Lozoya, Javier A1 - Lech, Maciej A1 - Adorjan, Kristina A1 - Stubbe, Hans Christian A1 - Hanssen, Henner A1 - Kotliar, Konstantin A1 - Haller, Berhard A1 - Heemann, Uwe A1 - Schmaderer, Christoph T1 - Persistent endothelial dysfunction in post-COVID-19 syndrome and its associations with symptom severity and chronic inflammation N2 - Background Post-COVID-19 syndrome (PCS) is a lingering disease with ongoing symptoms such as fatigue and cognitive impairment resulting in a high impact on the daily life of patients. Understanding the pathophysiology of PCS is a public health priority, as it still poses a diagnostic and treatment challenge for physicians. Methods In this prospective observational cohort study, we analyzed the retinal microcirculation using Retinal Vessel Analysis (RVA) in a cohort of patients with PCS and compared it to an age- and gender-matched healthy cohort (n = 41, matched out of n = 204). Measurements and main results PCS patients exhibit persistent endothelial dysfunction (ED), as indicated by significantly lower venular flicker-induced dilation (vFID; 3.42% ± 1.77% vs. 4.64% ± 2.59%; p = 0.02), narrower central retinal artery equivalent (CRAE; 178.1 [167.5–190.2] vs. 189.1 [179.4–197.2], p = 0.01) and lower arteriolar-venular ratio (AVR; (0.84 [0.8–0.9] vs. 0.88 [0.8–0.9], p = 0.007). When combining AVR and vFID, predicted scores reached good ability to discriminate groups (area under the curve: 0.75). Higher PCS severity scores correlated with lower AVR (R = − 0.37 p = 0.017). The association of microvascular changes with PCS severity were amplified in PCS patients exhibiting higher levels of inflammatory parameters. Conclusion Our results demonstrate that prolonged endothelial dysfunction is a hallmark of PCS, and impairments of the microcirculation seem to explain ongoing symptoms in patients. As potential therapies for PCS emerge, RVA parameters may become relevant as clinical biomarkers for diagnosis and therapy management. KW - Endothelial dysfunction KW - Long COVID KW - Post-COVID-19 syndrome KW - retinal microvasculature Y1 - 2023 U6 - http://dx.doi.org/10.1007/s10456-023-09885-6 N1 - Corresponding author: Christoph Schmaderer VL - 26 SP - 547 EP - 563 PB - Springer Nature CY - Dordrecht ER -