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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.
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.
In comparison to single-analyte devices, multiplexed systems for a multianalyte detection offer a reduced assay time and sample volume, low cost, and high throughput. Herein, a multiplexing platform for an automated quasi-simultaneous characterization of multiple (up to 16) capacitive field-effect sensors by the capacitive–voltage (C–V) and the constant-capacitance (ConCap) mode is presented. The sensors are mounted in a newly designed multicell arrangement with one common reference electrode and are electrically connected to the impedance analyzer via the base station. A Python script for the automated characterization of the sensors executes the user-defined measurement protocol. The developed multiplexing system is tested for pH measurements and the label-free detection of ligand-stabilized, charged gold nanoparticles.
With the prevalence of glucosamine- and chondroitin-containing dietary supplements for people with osteoarthritis in the marketplace, it is important to have an accurate and reproducible analytical method for the quantitation of these compounds in finished products. NMR spectroscopic method based both on low- (80 MHz) and high- (500–600 MHz) field NMR instrumentation was established, compared and validated for the determination of chondroitin sulfate and glucosamine in dietary supplements. The proposed method was applied for analysis of 20 different dietary supplements. In the majority of cases, quantification results obtained on the low-field NMR spectrometer are similar to those obtained with high-field 500–600 MHz NMR devices. Validation results in terms of accuracy, precision, reproducibility, limit of detection and recovery demonstrated that the developed method is fit for purpose for the marketed products. The NMR method was extended to the analysis of methylsulfonylmethane, adulterant maltodextrin, acetate and inorganic ions. Low-field NMR can be a quicker and cheaper alternative to more expensive high-field NMR measurements for quality control of the investigated dietary supplements. High-field NMR instrumentation can be more favorable for samples with complex composition due to better resolution, simultaneously giving the possibility of analysis of inorganic species such as potassium and chloride.
This work presents the Multi-Bees-Tracker (MBT3D) algorithm, a Python framework implementing a deep association tracker for Tracking-By-Detection, to address the challenging task of tracking flight paths of bumblebees in a social group. While tracking algorithms for bumblebees exist, they often come with intensive restrictions, such as the need for sufficient lighting, high contrast between the animal and background, absence of occlusion, significant user input, etc. Tracking flight paths of bumblebees in a social group is challenging. They suddenly adjust movements and change their appearance during different wing beat states while exhibiting significant similarities in their individual appearance. The MBT3D tracker, developed in this research, is an adaptation of an existing ant tracking algorithm for bumblebee tracking. It incorporates an offline trained appearance descriptor along with a Kalman Filter for appearance and motion matching. Different detector architectures for upstream detections (You Only Look Once (YOLOv5), Faster Region Proposal Convolutional Neural Network (Faster R-CNN), and RetinaNet) are investigated in a comparative study to optimize performance. The detection models were trained on a dataset containing 11359 labeled bumblebee images. YOLOv5 reaches an Average Precision of AP = 53, 8%, Faster R-CNN achieves AP = 45, 3% and RetinaNet AP = 38, 4% on the bumblebee validation dataset, which consists of 1323 labeled bumblebee images. The tracker’s appearance model is trained on 144 samples. The tracker (with Faster R-CNN detections) reaches a Multiple Object Tracking Accuracy MOTA = 93, 5% and a Multiple Object Tracking Precision MOTP = 75, 6% on a validation dataset containing 2000 images, competing with state-of-the-art computer vision methods. The framework allows reliable tracking of different bumblebees in the same video stream with rarely occurring identity switches (IDS). MBT3D has much lower IDS than other commonly used algorithms, with one of the lowest false positive rates, competing with state-of-the-art animal tracking algorithms. The developed framework reconstructs the 3-dimensional (3D) flight paths of the bumblebees by triangulation. It also handles and compares two alternative stereo camera pairs if desired.
Lead and nickel, as heavy metals, are still used in industrial processes, and are classified as “environmental health hazards” due to their toxicity and polluting potential. The detection of heavy metals can prevent environmental pollution at toxic levels that are critical to human health. In this sense, the electrolyte–insulator–semiconductor (EIS) field-effect sensor is an attractive sensing platform concerning the fabrication of reusable and robust sensors to detect such substances. This study is aimed to fabricate a sensing unit on an EIS device based on Sn₃O₄ nanobelts embedded in a polyelectrolyte matrix of polyvinylpyrrolidone (PVP) and polyacrylic acid (PAA) using the layer-by-layer (LbL) technique. The EIS-Sn₃O₄ sensor exhibited enhanced electrochemical performance for detecting Pb²⁺ and Ni²⁺ ions, revealing a higher affinity for Pb²⁺ ions, with sensitivities of ca. 25.8 mV/decade and 2.4 mV/decade, respectively. Such results indicate that Sn₃O₄ nanobelts can contemplate a feasible proof-of-concept capacitive field-effect sensor for heavy metal detection, envisaging other future studies focusing on environmental monitoring.
Kartellrecht vs. Datenschutzrecht: Rechtsgrundlagen für die Datenverarbeitung in sozialen Netzwerken
(2023)
Throughout the last decade, and particularly in 2022, water scarcity has become a critical concern in Morocco and other Mediterranean countries. The lack of rainfall during spring was worsened by a succession of heat waves during the summer. To address this drought, innovative solutions, including the use of new technologies such as hydrogels, will be essential to transform agriculture. This paper presents the findings of a study that evaluated the impact of hydrogel application on onion (Allium cepa) cultivation in Meknes, Morocco. The treatments investigated in this study comprised two different types of hydrogel-based soil additives (Arbovit® polyacrylate and Huminsorb® polyacrylate), applied at two rates (30 and 20 kg/ha), and irrigated at two levels of water supply (100% and 50% of daily crop evapotranspiration; ETc). Two control treatments were included, without hydrogel application and with both water amounts. The experiment was conducted in an open field using a completely randomized design. The results indicated a significant impact of both hydrogel-type dose and water dose on onion plant growth, as evidenced by various vegetation parameters. Among the hydrogels tested, Huminsorb® Polyacrylate produced the most favorable outcomes, with treatment T9 (100%, HP, 30 kg/ha) yielding 70.55 t/ha; this represented an increase of 11 t/ha as compared to the 100% ETc treatment without hydrogel application. Moreover, the combination of hydrogel application with 50% ETc water stress showed promising results, with treatment T4 (HP, 30 kg, 50%) producing almost the same yield as the 100% ETc treatment without hydrogel while saving 208 mm of water.
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.
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.
Today’s society is undergoing a paradigm shift driven by the megatrend of sustainability. This undeniably affects all areas of Western life. This paper aims to find out how the luxury industry is dealing with this change and what adjustments are made by the companies. For this purpose, interviews were conducted with managers from the luxury industry, in which they were asked about specific measures taken by their companies as well as trends in the industry. In a subsequent evaluation, the trends in the luxury industry were summarized for the areas of ecological, social, and economic sustainability. It was found that the area of environmental sustainability is significantly more focused than the other sub-areas. Furthermore, the need for a customer survey to validate the industry-based measures was identified.
Umsatzbasierte Bußgelder – wie sonst nur aus dem Kartellrecht bekannt – waren einer der Gründe, warum die Datenschutz-Grundverordnung (DSGVO) vor ihrem Inkrafttreten für erhebliches Aufsehen sorgte. Die vielfach relevanteren Schadensersatzansprüche, die, wie bei „Dieselgate“, aufgrund der Vielzahl von betroffenen Personen und der aus Sicht von Rechtsdienstleistern bestehenden Skalierbarkeit mit weitaus höheren Einbußen für Unternehmen einhergehen können, blieben zunächst unbeachtet. Inzwischen ist der Schadensersatzanspruch gem. Art. 82 DSGVO die Vorschrift, die die meisten Vorlagen zum Europäischen Gerichtshof (EuGH) der letzten Jahre hervorgerufen hat. Am 4.5.2023 hat nun der EuGH (Urteil v. 4.5.2023 - Rs. C-300/21, NWB GAAAJ-41389) in einem Grundsatzurteil über zentrale Fragen rund um den Ersatz immaterieller Schäden als Folge von Datenschutzverstößen entschieden.
The number of electronic vehicles increase steadily while the space for extending the charging infrastructure is limited. In particular in urban areas, where parking spaces in attractive areas are famous, opportunities to setup new charging stations is very limited. This leads to an overload of some very attractive charging stations and an underutilization of less attractive ones. Against this background, the paper at hand presents the design of an e-vehicle reservation system that aims at distributing the utilization of the charging infrastructure, particularly in urban areas. By applying a design science approach, the requirements for a reservation-based utilization approach are elicited and a model for a suitable distribution approach and its instantiation are developed. The artefact is evaluated by simulating the distribution effects based on data of real charging station utilizations.
New European Union (EU) regulations for UAS operations require an operational risk analysis, which includes an estimation of the potential danger of the UAS crashing. A key parameter for the potential ground risk is the kinetic impact energy of the UAS. The kinetic energy depends on the impact velocity of the UAS and, therefore, on the aerodynamic drag and the weight during free fall. Hence, estimating the impact energy of a UAS requires an accurate drag estimation of the UAS in that state. The paper at hand presents the aerodynamic drag estimation of small-scale multirotor UAS. Multirotor UAS of various sizes and configurations were analysed with a fully unsteady Reynolds-averaged Navier–Stokes approach. These simulations included different velocities and various fuselage pitch angles of the UAS. The results were compared against force measurements performed in a subsonic wind tunnel and provided good consistency. Furthermore, the influence of the UAS`s fuselage pitch angle as well as the influence of fixed and free spinning propellers on the aerodynamic drag was analysed. Free spinning propellers may increase the drag by up to 110%, depending on the fuselage pitch angle. Increasing the fuselage pitch angle of the UAS lowers the drag by 40% up to 85%, depending on the UAS. The data presented in this paper allow for increased accuracy of ground risk assessments.
Melting probes are a proven tool for the exploration of thick ice layers and clean sampling of subglacial water on Earth. Their compact size and ease of operation also make them a key technology for the future exploration of icy moons in our Solar System, most prominently Europa and Enceladus. For both mission planning and hardware engineering, metrics such as efficiency and expected performance in terms of achievable speed, power requirements, and necessary heating power have to be known.
Theoretical studies aim at describing thermal losses on the one hand, while laboratory experiments and field tests allow an empirical investigation of the true performance on the other hand. To investigate the practical value of a performance model for the operational performance in extraterrestrial environments, we first contrast measured data from terrestrial field tests on temperate and polythermal glaciers with results from basic heat loss models and a melt trajectory model. For this purpose, we propose conventions for the determination of two different efficiencies that can be applied to both measured data and models. One definition of efficiency is related to the melting head only, while the other definition considers the melting probe as a whole. We also present methods to combine several sources of heat loss for probes with a circular cross-section, and to translate the geometry of probes with a non-circular cross-section to analyse them in the same way. The models were selected in a way that minimizes the need to make assumptions about unknown parameters of the probe or the ice environment.
The results indicate that currently used models do not yet reliably reproduce the performance of a probe under realistic conditions. Melting velocities and efficiencies are constantly overestimated by 15 to 50 % in the models, but qualitatively agree with the field test data. Hence, losses are observed, that are not yet covered and quantified by the available loss models. We find that the deviation increases with decreasing ice temperature. We suspect that this mismatch is mainly due to the too restrictive idealization of the probe model and the fact that the probe was not operated in an efficiency-optimized manner during the field tests. With respect to space mission engineering, we find that performance and efficiency models must be used with caution in unknown ice environments, as various ice parameters have a significant effect on the melting process. Some of these are difficult to estimate from afar.
Subglacial environments on Earth offer important analogs to Ocean World targets in our solar system. These unique microbial ecosystems remain understudied due to the challenges of access through thick glacial ice (tens to hundreds of meters). Additionally, sub-ice collections must be conducted in a clean manner to ensure sample integrity for downstream microbiological and geochemical analyses. We describe the field-based cleaning of a melt probe that was used to collect brine samples from within a glacier conduit at Blood Falls, Antarctica, for geomicrobiological studies. We used a thermoelectric melting probe called the IceMole that was designed to be minimally invasive in that the logistical requirements in support of drilling operations were small and the probe could be cleaned, even in a remote field setting, so as to minimize potential contamination. In our study, the exterior bioburden on the IceMole was reduced to levels measured in most clean rooms, and below that of the ice surrounding our sampling target. Potential microbial contaminants were identified during the cleaning process; however, very few were detected in the final englacial sample collected with the IceMole and were present in extremely low abundances (∼0.063% of 16S rRNA gene amplicon sequences). This cleaning protocol can help minimize contamination when working in remote field locations, support microbiological sampling of terrestrial subglacial environments using melting probes, and help inform planetary protection challenges for Ocean World analog mission concepts.
Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.
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.