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Background
Aminoacylases are highly promising enzymes for the green synthesis of acyl-amino acids, potentially replacing the environmentally harmful Schotten-Baumann reaction. Long-chain acyl-amino acids can serve as strong surfactants and emulsifiers, with application in cosmetic industries. Heterologous expression of these enzymes, however, is often hampered, limiting their use in industrial processes.
Results
We identified a novel mycobacterial aminoacylase gene from Mycolicibacterium smegmatis MKD 8, cloned and expressed it in Escherichia coli and Vibrio natriegens using the T7 overexpression system. The recombinant enzyme was prone to aggregate as inclusion bodies, and while V. natriegens Vmax™ could produce soluble aminoacylase upon induction with isopropyl β-d-1-thiogalactopyranoside (IPTG), E. coli BL21 (DE3) needed autoinduction with lactose to produce soluble recombinant protein. We successfully conducted a chaperone co-expression study in both organisms to further enhance aminoacylase production and found that overexpression of chaperones GroEL/S enhanced aminoacylase activity in the cell-free extract 1.8-fold in V. natriegens and E. coli. Eventually, E. coli ArcticExpress™ (DE3), which co-expresses cold-adapted chaperonins Cpn60/10 from Oleispira antarctica, cultivated at 12 °C, rendered the most suitable expression system for this aminoacylase and exhibited twice the aminoacylase activity in the cell-free extract compared to E. coli BL21 (DE3) with GroEL/S co-expression at 20 °C. The purified aminoacylase was characterized based on hydrolytic activities, being most stable and active at pH 7.0, with a maximum activity at 70 °C, and stability at 40 °C and pH 7.0 for 5 days. The aminoacylase strongly prefers short-chain acyl-amino acids with smaller, hydrophobic amino acid residues. Several long-chain amino acids were fairly accepted in hydrolysis as well, especially N-lauroyl-L-methionine. To initially evaluate the relevance of this aminoacylase for the synthesis of N-acyl-amino acids, we demonstrated that lauroyl-methionine can be synthesized from lauric acid and methionine in an aqueous system.
Conclusion
Our results suggest that the recombinant enzyme is well suited for synthesis reactions and will thus be further investigated.
The eVTOL industry is a rapidly growing mass market expected to start in 2024. eVTOL compete, caused by their predicted missions, with ground-based transportation modes, including mainly passenger cars. Therefore, the automotive and classical aircraft design process is reviewed and compared to highlight advantages for eVTOL development. A special focus is on ergonomic comfort and safety. The need for further investigation of eVTOL’s crashworthiness is outlined by, first, specifying the relevance of passive safety via accident statistics and customer perception analysis; second, comparing the current state of regulation and certification; and third, discussing the advantages of integral safety and applying the automotive safety approach for eVTOL development. Integral safety links active and passive safety, while the automotive safety approach means implementing standardized mandatory full-vehicle crash tests for future eVTOL. Subsequently, possible crash impact conditions are analyzed, and three full-vehicle crash load cases are presented.
Die IMechE Railway Challenge wird jährlich in Stapleford, Großbritannien ausgetragen. Im Rahmen der Challenge entwickeln und bauen Studierende eine Lokomotive und vergleichen sich in verschiedenen Disziplinen, darunter eine automatisierte Zielbremsung, optimale Energierückgewinnung beim Bremsen und minimale Geräuschemissionen. Neben diesen und weiteren technischen Wettbewerbsdisziplinen treten die Fahrzeuge und die Teams auch in nicht-technischen Disziplinen wie einer Business Case Challenge an.
This study analyses the expected utilization of an urban distribution grid under high penetration of photovoltaic and e-mobility with charging infrastructure on a residential level. The grid utilization and the corresponding power flow are evaluated, while varying the control strategies and photovoltaic installed capacity in different scenarios. Four scenarios are used to analyze the impact of e-mobility. The individual mobility demand is modelled based on the largest German studies on mobility “Mobilität in Deutschland”, which is carried out every 5 years. To estimate the ramp-up of photovoltaic generation, a potential analysis of the roof surfaces in the supply area is carried out via an evaluation of an open solar potential study. The photovoltaic feed-in time series is derived individually for each installed system in a resolution of 15 min. The residential consumption is estimated using historical smart meter data, which are collected in London between 2012 and 2014. For a realistic charging demand, each residential household decides daily on the state of charge if their vehicle requires to be charged. The resulting charging time series depends on the underlying behavior scenario. Market prices and mobility demand are therefore used as scenario input parameters for a utility function based on the current state of charge to model individual behavior. The aggregated electricity demand is the starting point of the power flow calculation. The evaluation is carried out for an urban region with approximately 3100 residents. The analysis shows that increased penetration of photovoltaics combined with a flexible and adaptive charging strategy can maximize PV usage and reduce the need for congestion-related intervention by the grid operator by reducing the amount of kWh charged from the grid by 30% which reduces the average price of a charged kWh by 35% to 14 ct/kWh from 21.8 ct/kWh without PV optimization. The resulting grid congestions are managed by implementing an intelligent price or control signal. The analysis took place using data from a real German grid with 10 subgrids. The entire software can be adapted for the analysis of different distribution grids and is publicly available as an open-source software library on GitHub.
Environmental emissions, global warming, and energy-related concerns have accelerated the advancements in conventional vehicles that primarily use internal combustion engines. Among the existing technologies, hydrogen fuel cell electric vehicles and fuel cell hybrid electric vehicles may have minimal contributions to greenhouse gas emissions and thus are the prime choices for environmental concerns. However, energy management in fuel cell electric vehicles and fuel cell hybrid electric vehicles is a major challenge. Appropriate control strategies should be used for effective energy management in these vehicles. On the other hand, there has been significant progress in artificial intelligence, machine learning, and designing data-driven intelligent controllers. These techniques have found much attention within the community, and state-of-the-art energy management technologies have been developed based on them. This manuscript reviews the application of machine learning and intelligent controllers for prediction, control, energy management, and vehicle to everything (V2X) in hydrogen fuel cell vehicles. The effectiveness of data-driven control and optimization systems are investigated to evolve, classify, and compare, and future trends and directions for sustainability are discussed.
On the applicability of several tests to models with not identically distributed random effects
(2023)
We consider Kolmogorov–Smirnov and Cramér–von-Mises type tests for testing central symmetry, exchangeability, and independence. In the standard case, the tests are intended for the application to independent and identically distributed data with unknown distribution. The tests are available for multivariate data and bootstrap procedures are suitable to obtain critical values. We discuss the applicability of the tests to random effects models, where the random effects are independent but not necessarily identically distributed and with possibly unknown distributions. Theoretical results show the adequacy of the tests in this situation. The quality of the tests in models with random effects is investigated by simulations. Empirical results obtained confirm the theoretical findings. A real data example illustrates the application.
We consider time-dependent portfolios and discuss the allocation of changes in the risk of a portfolio to changes in the portfolio’s components. For this purpose we adopt established allocation principles. We also use our approach to obtain forecasts for changes in the risk of the portfolio’s components. To put the approach into practice we present an implementation based on the output of a simulation. Allocation is illustrated with an example portfolio in the context of Solvency II. The quality of the forecasts is investigated with an empirical study.
This article describes an Internet of things (IoT) sensing device with a wireless interface which is powered by the energy-harvesting method of the Wiegand effect. The Wiegand effect, in contrast to continuous sources like photovoltaic or thermal harvesters, provides small amounts of energy discontinuously in pulsed mode. To enable an energy-self-sufficient operation of the sensing device with this pulsed energy source, the output energy of the Wiegand generator is maximized. This energy is used to power up the system and to acquire and process data like position, temperature or other resistively measurable quantities as well as transmit these data via an ultra-low-power ultra-wideband (UWB) data transmitter. A proof-of-concept system was built to prove the feasibility of the approach. The energy consumption of the system during start-up was analysed, traced back in detail to the individual components, compared to the generated energy and processed to identify further optimization options. Based on the proof of concept, an application prototype was developed.
Motile cilia are hair-like cell extensions that beat periodically to generate fluid flow along various epithelial tissues within the body. In dense multiciliated carpets, cilia were shown to exhibit a remarkable coordination of their beat in the form of traveling metachronal waves, a phenomenon which supposedly enhances fluid transport. Yet, how cilia coordinate their regular beat in multiciliated epithelia to move fluids remains insufficiently understood, particularly due to lack of rigorous quantification. We combine experiments, novel analysis tools, and theory to address this knowledge gap. To investigate collective dynamics of cilia, we studied zebrafish multiciliated epithelia in the nose and the brain. We focused mainly on the zebrafish nose, due to its conserved properties with other ciliated tissues and its superior accessibility for non-invasive imaging. We revealed that cilia are synchronized only locally and that the size of local synchronization domains increases with the viscosity of the surrounding medium. Even though synchronization is local only, we observed global patterns of traveling metachronal waves across the zebrafish multiciliated epithelium. Intriguingly, these global wave direction patterns are conserved across individual fish, but different for left and right noses, unveiling a chiral asymmetry of metachronal coordination. To understand the implications of synchronization for fluid pumping, we used a computational model of a regular array of cilia. We found that local metachronal synchronization prevents steric collisions, i.e., cilia colliding with each other, and improves fluid pumping in dense cilia carpets, but hardly affects the direction of fluid flow. In conclusion, we show that local synchronization together with tissue-scale cilia alignment coincide and generate metachronal wave patterns in multiciliated epithelia, which enhance their physiological function of fluid pumping.
Assistance systems have been widely adopted in the manufacturing sector to facilitate various processes and tasks in production environments. However, existing systems are mostly equipped with rigid functional logic and do not provide individual user experiences or adapt to their capabilities. This work integrates human factors in assistance systems by adjusting the hardware and instruction presented to the workers’ cognitive and physical demands. A modular system architecture is designed accordingly, which allows a flexible component exchange according to the user and the work task. Gamification, the use of game elements in non-gaming contexts, has been further adopted in this work to provide level-based instructions and personalised feedback. The developed framework is validated by applying it to a manual workstation for industrial assembly routines.
High aerodynamic efficiency requires propellers with high aspect ratios, while propeller sweep potentially reduces noise. Propeller sweep and high aspect ratios increase elasticity and coupling of structural mechanics and aerodynamics, affecting the propeller performance and noise. Therefore, this paper analyzes the influence of elasticity on forward-swept, backward-swept, and unswept propellers in hover conditions. A reduced-order blade element momentum approach is coupled with a one-dimensional Timoshenko beam theory and Farassat's formulation 1A. The results of the aeroelastic simulation are used as input for the aeroacoustic calculation. The analysis shows that elasticity influences noise radiation because thickness and loading noise respond differently to deformations. In the case of the backward-swept propeller, the location of the maximum sound pressure level shifts forward by 0.5 °, while in the case of the forward-swept propeller, it shifts backward by 0.5 °. Therefore, aeroacoustic optimization requires the consideration of propeller deformation.
Next-generation aircraft designs often incorporate multiple large propellers attached along the wingspan. These highly flexible dynamic systems can exhibit uncommon aeroelastic instabilities, which should be carefully investigated to ensure safe operation. The interaction between the propeller and the wing is of particular importance. It is known that whirl flutter is stabilized by wing motion and wing aerodynamics. This paper investigates the effect of a propeller onto wing flutter as a function of span position and mounting stiffness between the propeller and wing. The analysis of a comparison between a tractor and pusher configuration has shown that the coupled system is more stable than the standalone wing for propeller positions near the wing tip for both configurations. The wing fluttermechanism is mostly affected by the mass of the propeller and the resulting change in eigenfrequencies of the wing. For very weak mounting stiffnesses, whirl flutter occurs, which was shown to be stabilized compared to a standalone propeller due to wing motion. On the other hand, the pusher configuration is, as to be expected, the more critical configuration due to the attached mass behind the elastic axis.
Based on the European Space Agency (ESA) Science in Space Environment (SciSpacE) community White Paper “Human Physiology – Musculoskeletal system”, this perspective highlights unmet needs and suggests new avenues for future studies in musculoskeletal research to enable crewed exploration missions. The musculoskeletal system is essential for sustaining physical function and energy metabolism, and the maintenance of health during exploration missions, and consequently mission success, will be tightly linked to musculoskeletal function. Data collection from current space missions from pre-, during-, and post-flight periods would provide important information to understand and ultimately offset musculoskeletal alterations during long-term spaceflight. In addition, understanding the kinetics of the different components of the musculoskeletal system in parallel with a detailed description of the molecular mechanisms driving these alterations appears to be the best approach to address potential musculoskeletal problems that future exploratory-mission crew will face. These research efforts should be accompanied by technical advances in molecular and phenotypic monitoring tools to provide in-flight real-time feedback.
The Cramér-von-Mises distance is applied to the distribution of the excess over a confidence level. Asymptotics of related statistics are investigated, and it is seen that the obtained limit distributions differ from the classical ones. For that reason, quantiles of the new limit distributions are given and new bootstrap techniques for approximation purposes are introduced and justified. The results motivate new one-sample goodness-of-fit tests for the distribution of the excess over a confidence level and a new confidence interval for the related fitting error. Simulation studies investigate size and power of the tests as well as coverage probabilities of the confidence interval in the finite sample case. A practice-oriented application of the Cramér-von-Mises tests is the determination of an appropriate confidence level for the fitting approach. The adoption of the idea to the well-known problem of threshold detection in the context of peaks over threshold modelling is sketched and illustrated by data examples.
In this paper, we provide an analytical study of the transmission eigenvalue problem with two conductivity parameters. We will assume that the underlying physical model is given by the scattering of a plane wave for an isotropic scatterer. In previous studies, this eigenvalue problem was analyzed with one conductive boundary parameter whereas we will consider the case of two parameters. We prove the existence and discreteness of the transmission eigenvalues as well as study the dependence on the physical parameters. We are able to prove monotonicity of the first transmission eigenvalue with respect to the parameters and consider the limiting procedure as the second boundary parameter vanishes. Lastly, we provide extensive numerical experiments to validate the theoretical work.
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.
Ambitious climate targets affect the competitiveness of industries in the international market. To prevent such industries from moving to other countries in the wake of increased climate protection efforts, cost adjustments may become necessary. Their design requires knowledge of country-specific production costs. Here, we present country-specific cost figures for different production routes of steel, paying particular attention to transportation costs. The data can be used in floor price models aiming to assess the competitiveness of different steel production routes in different countries (Rübbelke, 2022).
Optical Fibers as Dosimeter Detectors for Mixed Proton/Neutron Fields - A Biological Dosimeter
(2023)
In recent years, proton therapy has gained importance as a cancer treatment modality due to its conformality with the tumor and the sparing of healthy tissue. However, in the interaction of the protons with the beam line elements and patient tissues, potentially harmful secondary neutrons are always generated. To ensure that this neutron dose is as low as possible, treatment plans could be created to also account for and minimize the neutron dose. To monitor such a treatment plan, a compact, easy to use, and inexpensive dosimeter must be developed that not only measures the physical dose, but which can also distinguish between proton and neutron contributions. To that end, plastic optical fibers with scintillation materials (Gd₂O₂S:Tb, Gd₂O₂S:Eu, and YVO₄:Eu) were irradiated with protons and neutrons. It was confirmed that sensors with different scintillation materials have different sensitivities to protons and neutrons. A combination of these three scintillators can be used to build a detector array to create a biological dosimeter.
Influence of slab deflection on the out-of-plane capacity of unreinforced masonry partition walls
(2023)
Severe damage of non-structural elements is noticed in previous earthquakes, causing high economic losses and posing a life threat for the people. Masonry partition walls are one of the most commonly used non-structural elements. Therefore, their behaviour under earthquake loading in out-of-plane (OOP) direction is investigated by several researches in the past years. However, none of the existing experimental campaigns or analytical approaches consider the influence of prior slab deflection on OOP response of partition walls. Moreover, none of the existing construction techniques for the connection of partition walls with surrounding reinforced concrete (RC) is investigated for the combined slab deflection and OOP loading. However, the inevitable time-dependent behaviour of RC slabs leads to high values of final slab deflections which can further influence boundary conditions of partition walls. Therefore, a comprehensive study on the influence of slab deflection on the OOP capacity of masonry partitions is conducted. In the first step, experimental tests are carried out. Results of experimental tests are further used for the calibration of the numerical model employed for a parametric study. Based on the results, behaviour under combined loading for different construction techniques is explained. The results show that slab deflection leads either to severe damage or to a high reduction of OOP capacity. Existing practical solutions do not account for these effects. In this contribution, recommendations to overcome the problems of combined slab deflection and OOP loading on masonry partition walls are given. Possible interaction of in-plane (IP) loading, with the combined slab deflection and OOP loading on partition walls, is not investigated in this study.