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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.
Several species of (poly)saccharides and organic acids can be found often simultaneously in various biological matrices, e.g., fruits, plant materials, and biological fluids. The analysis of such matrices sometimes represents a challenging task. Using Aloe vera (A. vera) plant materials as an example, the performance of several spectro-scopic methods (80 MHz benchtop NMR, NIR, ATR-FTIR and UV–vis) for the simultaneous analysis of quality parameters of this plant material was compared. The determined parameters include (poly)saccharides such as aloverose, fructose and glucose as well as organic acids (malic, lactic, citric, isocitric, acetic, fumaric, benzoic and sorbic acids). 500 MHz NMR and high-performance liquid chromatography (HPLC) were used as the reference methods.
UV–vis data can be used only for identification of added preservatives (benzoic and sorbic acids) and drying agent (maltodextrin) and semiquantitative analysis of malic acid. NIR and MIR spectroscopies combined with multivariate regression can deliver more informative overview of A. vera extracts being able to additionally quantify glucose, aloverose, citric, isocitric, malic, lactic acids and fructose. Low-field NMR measurements can be used for the quantification of aloverose, glucose, malic, lactic, acetic, and benzoic acids. The benchtop NMR method was successfully validated in terms of robustness, stability, precision, reproducibility and limit of detection (LOD) and quantification (LOQ), respectively. All spectroscopic techniques are useful for the screening of (poly)saccharides and organic acids in plant extracts and should be applied according to its availability as well as information and confidence required for the specific analytical goal. Benchtop NMR spectroscopy seems to be the most feasible solution for quality control of A. vera products.
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.
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.
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.
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.
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.
Herein, fibroin, polylactide (PLA), and carbon are investigated for their suitability as biocompatible and biodegradable materials for amperometric biosensors. For this purpose, screen-printed carbon electrodes on the biodegradable substrates fibroin and PLA are modified with a glucose oxidase membrane and then encapsulated with the biocompatible material Ecoflex. The influence of different curing parameters of the carbon electrodes on the resulting biosensor characteristics is studied. The morphology of the electrodes is investigated by scanning electron microscopy, and the biosensor performance is examined by amperometric measurements of glucose (0.5–10 mM) in phosphate buffer solution, pH 7.4, at an applied potential of 1.2 V versus a Ag/AgCl reference electrode. Instead of Ecoflex, fibroin, PLA, and wound adhesive are tested as alternative encapsulation compounds: a series of swelling tests with different fibroin compositions, PLA, and Ecoflex has been performed before characterizing the most promising candidates by chronoamperometry. Therefore, the carbon electrodes are completely covered with the particular encapsulation material. Chronoamperometric measurements with H2O2 concentrations between 0.5 and 10 mM enable studying the leakage current behavior.
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.
Deammonification for nitrogen removal in municipal wastewater in temperate and cold climate zones is currently limited to the side stream of municipal wastewater treatment plants (MWWTP). This study developed a conceptual model of a mainstream deammonification plant, designed for 30,000 P.E., considering possible solutions corresponding to the challenging mainstream conditions in Germany. In addition, the energy-saving potential, nitrogen elimination performance and construction-related costs of mainstream deammonification were compared to a conventional plant model, having a single-stage activated sludge process with upstream denitrification. The results revealed that an additional treatment step by combining chemical precipitation and ultra-fine screening is advantageous prior the mainstream deammonification. Hereby chemical oxygen demand (COD) can be reduced by 80% so that the COD:N ratio can be reduced from 12 to 2.5. Laboratory experiments testing mainstream conditions of temperature (8–20°C), pH (6–9) and COD:N ratio (1–6) showed an achievable volumetric nitrogen removal rate (VNRR) of at least 50 gN/(m3∙d) for various deammonifying sludges from side stream deammonification systems in the state of North Rhine-Westphalia, Germany, where m3 denotes reactor volume. Assuming a retained Norganic content of 0.0035 kgNorg./(P.E.∙d) from the daily loads of N at carbon removal stage and a VNRR of 50 gN/(m3∙d) under mainstream conditions, a resident-specific reactor volume of 0.115 m3/(P.E.) is required for mainstream deammonification. This is in the same order of magnitude as the conventional activated sludge process, i.e., 0.173 m3/(P.E.) for an MWWTP of size class of 4. The conventional plant model yielded a total specific electricity demand of 35 kWh/(P.E.∙a) for the operation of the whole MWWTP and an energy recovery potential of 15.8 kWh/(P.E.∙a) through anaerobic digestion. In contrast, the developed mainstream deammonification model plant would require only a 21.5 kWh/(P.E.∙a) energy demand and result in 24 kWh/(P.E.∙a) energy recovery potential, enabling the mainstream deammonification model plant to be self-sufficient. The retrofitting costs for the implementation of mainstream deammonification in existing conventional MWWTPs are nearly negligible as the existing units like activated sludge reactors, aerators and monitoring technology are reusable. However, the mainstream deammonification must meet the performance requirement of VNRR of about 50 gN/(m3∙d) in this case.
A method for detecting and approximating fault lines or surfaces, respectively, or decision curves in two and three dimensions with guaranteed accuracy is presented. Reformulated as a classification problem, our method starts from a set of scattered points along with the corresponding classification algorithm to construct a representation of a decision curve by points with prescribed maximal distance to the true decision curve. Hereby, our algorithm ensures that the representing point set covers the decision curve in its entire extent and features local refinement based on the geometric properties of the decision curve. We demonstrate applications of our method to problems related to the detection of faults, to multi-criteria decision aid and, in combination with Kirsch’s factorization method, to solving an inverse acoustic scattering problem. In all applications we considered in this work, our method requires significantly less pointwise classifications than previously employed algorithms.
The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465ᵀ and Alkalibacillus haloalkaliphilus DSM 5271ᵀ and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976ᵀ served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA. The protease genes were cloned and expressed in Bacillus subtilis DB104. Purification to apparent homogeneity was achieved by ethanol precipitation, desalting and ion-exchange chromatography. Enzyme activity could be observed between pH 5.0–12.0 with an optimum for SPPM, SPMI and SPLA around pH 9.0 and for SPAH at pH 10.0. The optimal temperature for SPMI and SPLA was 70 °C and for SPPM and SPAH 55 °C and 50 °C, respectively. All proteases showed high stability towards 5% (w/v) SDS and were active even at NaCl concentrations of 5 M. The four proteases demonstrate potential for future biotechnological applications.
Supervised machine learning and deep learning require a large amount of labeled data, which data scientists obtain in a manual, and time-consuming annotation process. To mitigate this challenge, Active Learning (AL) proposes promising data points to annotators they annotate next instead of a subsequent or random sample. This method is supposed to save annotation effort while maintaining model performance.
However, practitioners face many AL strategies for different tasks and need an empirical basis to choose between them. Surveys categorize AL strategies into taxonomies without performance indications. Presentations of novel AL strategies compare the performance to a small subset of strategies. Our contribution addresses the empirical basis by introducing a reproducible active learning evaluation (ALE) framework for the comparative evaluation of AL strategies in NLP.
The framework allows the implementation of AL strategies with low effort and a fair data-driven comparison through defining and tracking experiment parameters (e.g., initial dataset size, number of data points per query step, and the budget). ALE helps practitioners to make more informed decisions, and researchers can focus on developing new, effective AL strategies and deriving best practices for specific use cases. With best practices, practitioners can lower their annotation costs. We present a case study to illustrate how to use the framework.
The work in modern open-pit and underground mines requires the transportation of large amounts of resources between fixed points. The navigation to these fixed points is a repetitive task that can be automated. The challenge in automating the navigation of vehicles commonly used in mines is the systemic properties of such vehicles. Many mining vehicles, such as the one we have used in the research for this paper, use steering systems with an articulated joint bending the vehicle’s drive axis to change its course and a hydraulic drive system to actuate axial drive components or the movements of tippers if available. To address the difficulties of controlling such a vehicle, we present a model-predictive approach for controlling the vehicle. While the control optimisation based on a parallel error minimisation of the predicted state has already been established in the past, we provide insight into the design and implementation of an MPC for an articulated mining vehicle and show the results of real-world experiments in an open-pit mine environment.
Extracting workflow nets from textual descriptions can be used to simplify guidelines or formalize textual descriptions of formal processes like business processes and algorithms. The task of manually extracting processes, however, requires domain expertise and effort. While automatic process model extraction is desirable, annotating texts with formalized process models is expensive. Therefore, there are only a few machine-learning-based extraction approaches. Rule-based approaches, in turn, require domain specificity to work well and can rarely distinguish relevant and irrelevant information in textual descriptions. In this paper, we present GUIDO, a hybrid approach to the process model extraction task that first, classifies sentences regarding their relevance to the process model, using a BERT-based sentence classifier, and second, extracts a process model from the sentences classified as relevant, using dependency parsing. The presented approach achieves significantly better resul ts than a pure rule-based approach. GUIDO achieves an average behavioral similarity score of 0.93. Still, in comparison to purely machine-learning-based approaches, the annotation costs stay low.
In recent years, the development of large pretrained language models, such as BERT and GPT, significantly improved information extraction systems on various tasks, including relation classification. State-of-the-art systems are highly accurate on scientific benchmarks. A lack of explainability is currently a complicating factor in many real-world applications. Comprehensible systems are necessary to prevent biased, counterintuitive, or harmful decisions.
We introduce semantic extents, a concept to analyze decision patterns for the relation classification task. Semantic extents are the most influential parts of texts concerning classification decisions. Our definition allows similar procedures to determine semantic extents for humans and models. We provide an annotation tool and a software framework to determine semantic extents for humans and models conveniently and reproducibly. Comparing both reveals that models tend to learn shortcut patterns from data. These patterns are hard to detect with current interpretability methods, such as input reductions. Our approach can help detect and eliminate spurious decision patterns during model development. Semantic extents can increase the reliability and security of natural language processing systems. Semantic extents are an essential step in enabling applications in critical areas like healthcare or finance. Moreover, our work opens new research directions for developing methods to explain deep learning models.
Preprint: Studies on the enzymatic reduction of levulinic acid using Chiralidon-R and Chiralidon-S
(2023)
The enzymatic reduction of levulinic acid by the chiral catalysts Chiralidon-R and Chiralidon-S which are commercially available superabsorbed alcohol dehydrogenases is described. The Chiralidon®-R/S reduces the levulinic acid to the (R,S)-4-hydroxy valeric acid and the (R)- or (S)- gamma-valerolactone.
Like all preceding transformations of the manufacturing industry, the large-scale usage of production data will reshape the role of humans within the sociotechnical production ecosystem. To ensure that this transformation creates work systems in which employees are empowered, productive, healthy, and motivated, the transformation must be guided by principles of and research on human-centered work design. Specifically, measures must be taken at all levels of work design, ranging from (1) the work tasks to (2) the working conditions to (3) the organizational level and (4) the supra-organizational level. We present selected research across all four levels that showcase the opportunities and requirements that surface when striving for human-centered work design for the Internet of Production (IoP). (1) On the work task level, we illustrate the user-centered design of human-robot collaboration (HRC) and process planning in the composite industry as well as user-centered design factors for cognitive assistance systems. (2) On the working conditions level, we present a newly developed framework for the classification of HRC workplaces. (3) Moving to the organizational level, we show how corporate data can be used to facilitate best practice sharing in production networks, and we discuss the implications of the IoP for new leadership models. Finally, (4) on the supra-organizational level, we examine overarching ethical dimensions, investigating, e.g., how the new work contexts affect our understanding of responsibility and normative values such as autonomy and privacy. Overall, these interdisciplinary research perspectives highlight the importance and necessary scope of considering the human factor in the IoP.