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Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO₂-Ta₂O₅ layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1–3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta₂O₅-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate.
Bacterial cellulose (BC) is a biopolymer produced by different microorganisms, but in biotechnological practice, Komagataeibacter xylinus is used. The micro- and nanofibrillar structure of BC, which forms many different-sized pores, creates prerequisites for the introduction of other polymers into it, including those synthesized by other microorganisms. The study aims to develop a cocultivation system of BC and prebiotic producers to obtain BC-based composite material with prebiotic activity. In this study, pullulan (PUL) was found to stimulate the growth of the probiotic strain Lactobacillus rhamnosus GG better than the other microbial polysaccharides gellan and xanthan. BC/PUL biocomposite with prebiotic properties was obtained by cocultivation of Komagataeibacter xylinus and Aureobasidium pullulans, BC and PUL producers respectively, on molasses medium. The inclusion of PUL in BC is proved gravimetrically by scanning electron microscopy and by Fourier transformed infrared spectroscopy. Cocultivation demonstrated a composite effect on the aggregation and binding of BC fibers, which led to a significant improvement in mechanical properties. The developed approach for “grafting” of prebiotic activity on BC allows preparation of environmentally friendly composites of better quality.
We present a concise mini overview on the approaches to the disposal of nuclear waste currently used or deployed. The disposal of nuclear waste is the end point of nuclear waste management (NWM) activities and is the emplacement of waste in an appropriate facility without the intention to retrieve it. The IAEA has developed an internationally accepted classification scheme based on the end points of NWM, which is used as guidance. Retention times needed for safe isolation of waste radionuclides are estimated based on the radiotoxicity of nuclear waste. Disposal facilities usually rely on a multi-barrier defence system to isolate the waste from the biosphere, which comprises the natural geological barrier and the engineered barrier system. Disposal facilities could be of a trench type, vaults, tunnels, shafts, boreholes, or mined repositories. A graded approach relates the depth of the disposal facilities’ location with the level of hazard. Disposal practices demonstrate the reliability of nuclear waste disposal with minimal expected impacts on the environment and humans.
A capacitive electrolyte-insulator-semiconductor (EISCAP) biosensor modified with Tobacco mosaic virus (TMV) particles for the detection of acetoin is presented. The enzyme acetoin reductase (AR) was immobilized on the surface of the EISCAP using TMV particles as nanoscaffolds. The study focused on the optimization of the TMV-assisted AR immobilization on the Ta 2 O 5 -gate EISCAP surface. The TMV-assisted acetoin EISCAPs were electrochemically characterized by means of leakage-current, capacitance-voltage, and constant-capacitance measurements. The TMV-modified transducer surface was studied via scanning electron microscopy.
Acetoin and diacetyl have a major impact on the flavor of alcoholic beverages such as wine or beer. Therefore, their measurement is important during the fermentation process. Until now, gas chromatographic techniques have typically been applied; however, these require expensive laboratory equipment and trained staff, and do not allow for online monitoring. In this work, a capacitive electrolyte–insulator–semiconductor sensor modified with tobacco mosaic virus (TMV) particles as enzyme nanocarriers for the detection of acetoin and diacetyl is presented. The enzyme acetoin reductase from Alkalihalobacillus clausii DSM 8716ᵀ is immobilized via biotin–streptavidin affinity, binding to the surface of the TMV particles. The TMV-assisted biosensor is electrochemically characterized by means of leakage–current, capacitance–voltage, and constant capacitance measurements. In this paper, the novel biosensor is studied regarding its sensitivity and long-term stability in buffer solution. Moreover, the TMV-assisted capacitive field-effect sensor is applied for the detection of diacetyl for the first time. The measurement of acetoin and diacetyl with the same sensor setup is demonstrated. Finally, the successive detection of acetoin and diacetyl in buffer and in diluted beer is studied by tuning the sensitivity of the biosensor using the pH value of the measurement solution.
An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.
The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.
An interdisciplinary view on humane interfaces for digital shadows in the internet of production
(2022)
Digital shadows play a central role for the next generation industrial internet, also known as Internet of Production (IoP). However, prior research has not considered systematically how human actors interact with digital shadows, shaping their potential for success. To address this research gap, we assembled an interdisciplinary team of authors from diverse areas of human-centered research to propose and discuss design and research recommendations for the implementation of industrial user interfaces for digital shadows, as they are currently conceptualized for the IoP. Based on the four use cases of decision support systems, knowledge sharing in global production networks, human-robot collaboration, and monitoring employee workload, we derive recommendations for interface design and enhancing workers’ capabilities. This analysis is extended by introducing requirements from the higher-level perspectives of governance and organization.
The seismic performance and safety of major European industrial facilities has a global interest for Europe, its citizens and economy. A potential major disaster at an industrial site could affect several countries, probably far beyond the country where it is located. However, the seismic design and safety assessment of these facilities is practically based on national, often outdated seismic hazard assessment studies, due to many reasons, including the absence of a reliable, commonly developed seismic hazard model for whole Europe. This important gap is no more existing, as the 2020 European Seismic Hazard Model ESHM20 was released in December 2021. In this paper we investigate the expected impact of the adoption of ESHM20 on the seismic demand for industrial facilities, through the comparison of the ESHM20 probabilistic hazard at the sites where industrial facilities are located with the respective national and European regulations. The goal of this preliminary work in the framework of Working Group 13 of the European Association for Earthquake Engineering (EAEE), is to identify potential inadequacies in the design and safety control of existing industrial facilities and to highlight the expected impact of the adoption of the new European Seismic Hazard Model on the design of new industrial facilities and the safety assessment of existing ones.
Masonry infill walls are the most traditional enclosure system that is still widely used in RC frame buildings all over the world, particularly in seismic active regions. Although infill walls are usually neglected in seismic design, during an earthquake event they are subjected to in-plane and out-of-plane forces that can act separately or simultaneously. Since observations of damage to buildings after recent earthquakes showed detrimental effects of in-plane and out-of-plane load interaction on infill walls, the number of studies that focus on influence of in-plane damage on out-of-plane response has significantly increased. However, most of the xperimental campaigns have considered only solid infills and there is a lack of combined in-plane and out-of-plane experimental tests on masonry infills with openings, although windows and doors strongly affect seismic performance. In this paper, two types of experimental tests on infills with window openings are presented. The first is a pure out-of-plane test and the second one is a sequential in-plane and out-of-plane test aimed at investigating the effects of existing in-plane damage on outof-plane response. Additionally, findings from two tests with similar load procedure that were carried out on fully infilled RC frames in the scope of the same project are used for comparison. Test results clearly show that window opening increased vulnerability of infills to combined seismic actions and that prevention of damage in infills with openings is of the utmost importance for seismic safety.