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Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822ᵀ (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5% (w/v) SDS and 5% H₂O₂ (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H₂O₂, suggest it has potential for biotechnological applications.
The present work aimed to study the mainstream feasibility of the deammonifying sludge of side stream of municipal wastewater treatment plant (MWWTP) in Kaster, Germany. For this purpose, the deammonifying sludge available at the side stream was investigated for nitrogen (N) removal with respect to the operational factors temperature (15–30°C), pH value (6.0–8.0) and chemical oxygen demand (COD)/N ratio (≤1.5–6.0). The highest and lowest N-removal rates of 0.13 and 0.045 kg/(m³ d) are achieved at 30 and 15°C, respectively. Different conditions of pH and COD/N ratios in the SBRs of Partial nitritation/anammox (PN/A) significantly influenced both the metabolic processes and associated N-removal rates. The scientific insights gained from the current work signifies the possibility of mainstream PN/A at WWTPs. The current study forms a solid basis of operational window for the upcoming semi-technical trails to be conducted prior to the full-scale mainstream PN/A at WWTP Kaster and WWTPs globally.
Even the shortest flight through unknown, cluttered environments requires reliable local path planning algorithms to avoid unforeseen obstacles. The algorithm must evaluate alternative flight paths and identify the best path if an obstacle blocks its way. Commonly, weighted sums are used here. This work shows that weighted Chebyshev distances and factorial achievement scalarising functions are suitable alternatives to weighted sums if combined with the 3DVFH* local path planning algorithm. Both methods considerably reduce the failure probability of simulated flights in various environments. The standard 3DVFH* uses a weighted sum and has a failure probability of 50% in the test environments. A factorial achievement scalarising function, which minimises the worst combination of two out of four objective functions, reaches a failure probability of 26%; A weighted Chebyshev distance, which optimises the worst objective, has a failure probability of 30%. These results show promise for further enhancements and to support broader applicability.
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.
The increasing share of renewable electricity in the grid drives the need for sufficient storage capacity. Especially for seasonal storage, power-to-gas can be a promising approach. Biologically produced methane from hydrogen produced from surplus electricity can be used to substitute natural gas in the existing infrastructure. Current reactor types are not or are poorly optimized for flexible methanation. Therefore, this work proposes a new reactor type with a plug flow reactor (PFR) design. Simulations in COMSOL Multiphysics ® showed promising properties for operation in laminar flow. An experiment was conducted to support the simulation results and to determine the gas fraction of the novel reactor, which was measured to be 29%. Based on these simulations and experimental results, the reactor was constructed as a 14 m long, 50 mm diameter tube with a meandering orientation. Data processing was established, and a step experiment was performed. In addition, a kLa of 1 h−1 was determined. The results revealed that the experimental outcomes of the type of flow and gas fractions are in line with the theoretical simulation. The new design shows promising properties for flexible methanation and will be tested.
Automated driving is now possible in diverse road and traffic conditions. However, there are still situations that automated vehicles cannot handle safely and efficiently. In this case, a Transition of Control (ToC) is necessary so that the driver takes control of the driving. Executing a ToC requires the driver to get full situation awareness of the driving environment. If the driver fails to get back the control in a limited time, a Minimum Risk Maneuver (MRM) is executed to bring the vehicle into a safe state (e.g., decelerating to full stop). The execution of ToCs requires some time and can cause traffic disruption and safety risks that increase if several vehicles execute ToCs/MRMs at similar times and in the same area. This study proposes to use novel C-ITS traffic management measures where the infrastructure exploits V2X communications to assist Connected and Automated Vehicles (CAVs) in the execution of ToCs. The infrastructure can suggest a spatial distribution of ToCs, and inform vehicles of the locations where they could execute a safe stop in case of MRM. This paper reports the first field operational tests that validate the feasibility and quantify the benefits of the proposed infrastructure-assisted ToC and MRM management. The paper also presents the CAV and roadside infrastructure prototypes implemented and used in the trials. The conducted field trials demonstrate that infrastructure-assisted traffic management solutions can reduce safety risks and traffic disruptions.
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.
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.
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.