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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.
Hotelling’s T² tests in paired and independent survey samples are compared using the traditional asymptotic efficiency concepts of Hodges–Lehmann, Bahadur and Pitman, as well as through criteria based on the volumes of corresponding confidence regions. Conditions characterizing the superiority of a procedure are given in terms of population canonical correlation type coefficients. Statistical tests for checking these conditions are developed. Test statistics based on the eigenvalues of a symmetrized sample cross-covariance matrix are suggested, as well as test statistics based on sample canonical correlation type coefficients.
This paper considers a paired data framework and discusses the question of marginal homogeneity of bivariate high-dimensional or functional data. The related testing problem can be endowed into a more general setting for paired random variables taking values in a general Hilbert space. To address this problem, a Cramér–von-Mises type test statistic is applied and a bootstrap procedure is suggested to obtain critical values and finally a consistent test. The desired properties of a bootstrap test can be derived that are asymptotic exactness under the null hypothesis and consistency under alternatives. Simulations show the quality of the test in the finite sample case. A possible application is the comparison of two possibly dependent stock market returns based on functional data. The approach is demonstrated based on historical data for different stock market indices.
The European Union's aim to become climate neutral by 2050 necessitates ambitious efforts to reduce carbon emissions. Large reductions can be attained particularly in energy intensive sectors like iron and steel. In order to prevent the relocation of such industries outside the EU in the course of tightening environmental regulations, the establishment of a climate club jointly with other large emitters and alternatively the unilateral implementation of an international cross-border carbon tax mechanism are proposed. This article focuses on the latter option choosing the steel sector as an example. In particular, we investigate the financial conditions under which a European cross border mechanism is capable to protect hydrogen-based steel production routes employed in Europe against more polluting competition from abroad. By using a floor price model, we assess the competitiveness of different steel production routes in selected countries. We evaluate the climate friendliness of steel production on the basis of specific GHG emissions. In addition, we utilize an input-output price model. It enables us to assess impacts of rising cost of steel production on commodities using steel as intermediates. Our results raise concerns that a cross-border tax mechanism will not suffice to bring about competitiveness of hydrogen-based steel production in Europe because the cost tends to remain higher than the cost of steel production in e.g. China. Steel is a classic example for a good used mainly as intermediate for other products. Therefore, a cross-border tax mechanism for steel will increase the price of products produced in the EU that require steel as an input. This can in turn adversely affect competitiveness of these sectors. Hence, the effects of higher steel costs on European exports should be borne in mind and could require the cross-border adjustment mechanism to also subsidize exports.
Am Beispiel der Telekommunikationsindustrie zeigt der Beitrag eine konkrete Ausgestaltung anwendungsorientierter Forschung, die sowohl für die Praxis als auch für die Wissenschaft nutzen- und erkenntnisbringend ist. Forschungsgegenstand sind die Referenzmodelle des Industriegremiums TM Forum, die von vielen Telekommunikationsunternehmen zur Transformation ihrer Strukturen und Systeme genutzt werden. Es wird die langjährige Forschungstätigkeit bei der Weiterentwicklung und Anwendung dieser Referenzmodelle beschrieben. Dabei wird ein konsequent gestaltungsorientierter Forschungsansatz verfolgt. Das Zusammenspiel aus kontinuierlicher Weiterentwicklung in Zusammenarbeit mit einem Industriegremium und der Anwendung in vielfältigen Praxisprojekten führt zu einer erfolgreichen Symbiose aus praktischer Nutzengenerierung sowie wissenschaftlichem Erkenntnisgewinn. Der Beitrag stellt den gewählten Forschungsansatz anhand konkreter Beispiele vor. Darauf basierend werden Empfehlungen und Herausforderungen für eine gestaltungs- und praxisorientierte Forschung diskutiert.
Das Gesundheitswesen ist konfrontiert mit steigenden Kosten und einer immer schwieriger werdenden Personalsituation. Zeitgleich versprechen moderne Sprachsteuerungssysteme Prozesse in Arztpraxen und Krankenhäusern zu verschlanken und Vorgänge zu beschleunigen. Dennoch wird derzeit der Einsatz von Sprachsteuerungssystemen in Arztpraxen oder Krankenhäusern nur selten beobachtet, was auch an den besonders strengen Datenschutzauflagen der Datenschutzgrundverordnung (DSGVO) liegt. Darüber hinaus wirft die niedrige Nutzungsrate die Frage nach den konkreten Anforderungen und ihrer Umsetzbarkeit auf, was durch den vorliegenden Beitrag adressiert wird, indem die Ergebnisse von Interviews mit acht medizinischen Fachexperten ausgewertet werden. Ergänzend wird die technische Umsetzbarkeit einzelner Anforderungen mit unterschiedlichen Cloud-Anbietern erprobt.
We study the possibility to fabricate an arbitrary phase mask in a one-step laser-writing process inside the volume of an optical glass substrate. We derive the phase mask from a Gerchberg–Saxton-type algorithm as an array and create each individual phase shift using a refractive index modification of variable axial length. We realize the variable axial length by superimposing refractive index modifications induced by an ultra-short pulsed laser at different focusing depth. Each single modification is created by applying 1000 pulses with 15 μJ pulse energy at 100 kHz to a fixed spot of 25 μm diameter and the focus is then shifted axially in steps of 10 μm. With several proof-of-principle examples, we show the feasibility of our method. In particular, we identify the induced refractive index change to about a value of Δn=1.5⋅10−3. We also determine our current limitations by calculating the overlap in the form of a scalar product and we discuss possible future improvements.
This study addresses a proof-of-concept experiment with a biocompatible screen-printed carbon electrode deposited onto a biocompatible and biodegradable substrate, which is made of fibroin, a protein derived from silk of the Bombyx mori silkworm. To demonstrate the sensor performance, the carbon electrode is functionalized as a glucose biosensor with the enzyme glucose oxidase and encapsulated with a silicone rubber to ensure biocompatibility of the contact wires. The carbon electrode is fabricated by means of thick-film technology including a curing step to solidify the carbon paste. The influence of the curing temperature and curing time on the electrode morphology is analyzed via scanning electron microscopy. The electrochemical characterization of the glucose biosensor is performed by amperometric/voltammetric measurements of different glucose concentrations in phosphate buffer. Herein, systematic studies at applied potentials from 500 to 1200 mV to the carbon working electrode (vs the Ag/AgCl reference electrode) allow to determine the optimal working potential. Additionally, the influence of the curing parameters on the glucose sensitivity is examined over a time period of up to 361 days. The sensor shows a negligible cross-sensitivity toward ascorbic acid, noradrenaline, and adrenaline. The developed biocompatible biosensor is highly promising for future in vivo and epidermal applications.
Edge-based and face-based smoothed finite element methods (ES-FEM and FS-FEM, respectively) are modified versions of the finite element method allowing to achieve more accurate results and to reduce sensitivity to mesh distortion, at least for linear elements. These properties make the two methods very attractive. However, their implementation in a standard finite element code is nontrivial because it requires heavy and extensive modifications to the code architecture. In this article, we present an element-based formulation of ES-FEM and FS-FEM methods allowing to implement the two methods in a standard finite element code with no modifications to its architecture. Moreover, the element-based formulation permits to easily manage any type of element, especially in 3D models where, to the best of the authors' knowledge, only tetrahedral elements are used in FS-FEM applications found in the literature. Shape functions for non-simplex 3D elements are proposed in order to apply FS-FEM to any standard finite element.
Image reconstruction analysis for positron emission tomography with heterostructured scintillators
(2022)
The concept of structure engineering has been proposed for exploring the next generation of radiation detectors with improved performance. A TOF-PET geometry with heterostructured scintillators with a pixel size of 3.0×3.1×15 mm3 was simulated using Monte Carlo. The heterostructures consisted of alternating layers of BGO as a dense material with high stopping power and plastic (EJ232) as a fast light emitter. The detector time resolution was calculated as a function of the deposited and shared energy in both materials on an event-by-event basis. While sensitivity was reduced to 32% for 100 μm thick plastic layers and 52% for 50 μm, the CTR distribution improved to 204±49 ps and 220±41 ps respectively, compared to 276 ps that we considered for bulk BGO. The complex distribution of timing resolutions was accounted for in the reconstruction. We divided the events into three groups based on their CTR and modeled them with different Gaussian TOF kernels. On a NEMA IQ phantom, the heterostructures had better contrast recovery in early iterations. On the other hand, BGO achieved a better contrast to noise ratio (CNR) after the 15th iteration due to the higher sensitivity. The developed simulation and reconstruction methods constitute new tools for evaluating different detector designs with complex time responses.
This study investigated the anaerobic digestion of an algal–bacterial biofilm grown in artificial wastewater in an Algal Turf Scrubber (ATS). The ATS system was located in a greenhouse (50°54′19ʺN, 6°24′55ʺE, Germany) and was exposed to seasonal conditions during the experiment period. The methane (CH4) potential of untreated algal–bacterial biofilm (UAB) and thermally pretreated biofilm (PAB) using different microbial inocula was determined by anaerobic batch fermentation. Methane productivity of UAB differed significantly between microbial inocula of digested wastepaper, a mixture of manure and maize silage, anaerobic sewage sludge, and percolated green waste. UAB using sewage sludge as inoculum showed the highest methane productivity. The share of methane in biogas was dependent on inoculum. Using PAB, a strong positive impact on methane productivity was identified for the digested wastepaper (116.4%) and a mixture of manure and maize silage (107.4%) inocula. By contrast, the methane yield was significantly reduced for the digested anaerobic sewage sludge (50.6%) and percolated green waste (43.5%) inocula. To further evaluate the potential of algal–bacterial biofilm for biogas production in wastewater treatment and biogas plants in a circular bioeconomy, scale-up calculations were conducted. It was found that a 0.116 km2 ATS would be required in an average municipal wastewater treatment plant which can be viewed as problematic in terms of space consumption. However, a substantial amount of energy surplus (4.7–12.5 MWh a−1) can be gained through the addition of algal–bacterial biomass to the anaerobic digester of a municipal wastewater treatment plant. Wastewater treatment and subsequent energy production through algae show dominancy over conventional technologies.
Plant viruses are major contributors to crop losses and induce high economic costs worldwide. For reliable, on-site and early detection of plant viral diseases, portable biosensors are of great interest. In this study, a field-effect SiO2-gate electrolyte-insulator-semiconductor (EIS) sensor was utilized for the label-free electrostatic detection of tobacco mosaic virus (TMV) particles as a model plant pathogen. The capacitive EIS sensor has been characterized regarding its TMV sensitivity by means of constant-capacitance method. The EIS sensor was able to detect biotinylated TMV particles from a solution with a TMV concentration as low as 0.025 nM. A good correlation between the registered EIS sensor signal and the density of adsorbed TMV particles assessed from scanning electron microscopy images of the SiO2-gate chip surface was observed. Additionally, the isoelectric point of the biotinylated TMV particles was determined via zeta potential measurements and the influence of ionic strength of the measurement solution on the TMV-modified EIS sensor signal has been studied.
A new functionalization method to modify capacitive electrolyte–insulator–semiconductor (EIS) structures with nanofilms is presented. Layers of polyallylamine hydrochloride (PAH) and graphene oxide (GO) with the compound polyaniline:poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (PANI:PAAMPSA) are deposited onto a p-Si/SiO2 chip using the layer-by-layer technique (LbL). Two different enzymes (urease and penicillinase) are separately immobilized on top of a five-bilayer stack of the PAH:GO/PANI:PAAMPSA-modified EIS chip, forming a biosensor for detection of urea and penicillin, respectively. Electrochemical characterization is performed by constant capacitance (ConCap) measurements, and the film morphology is characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). An increase in the average sensitivity of the modified biosensors (EIS–nanofilm–enzyme) of around 15% is found in relation to sensors, only carrying the enzyme but without the nanofilm (EIS–enzyme). In this sense, the nanofilm acts as a stable bioreceptor onto the EIS chip improving the output signal in terms of sensitivity and stability.
Miniaturized electrolyte–insulator–semiconductor capacitors (EISCAPs) with ultrathin gate insulators have been studied in terms of their pH-sensitive sensor characteristics: three different EISCAP systems consisting of Al–p-Si–Ta2O5(5 nm), Al–p-Si–Si3N4(1 or 2 nm)–Ta2O5 (5 nm), and Al–p-Si–SiO2(3.6 nm)–Ta2O5(5 nm) layer structures are characterized in buffer solution with different pH values by means of capacitance–voltage and constant capacitance method. The SiO2 and Si3N4 gate insulators are deposited by rapid thermal oxidation and rapid thermal nitridation, respectively, whereas the Ta2O5 film is prepared by atomic layer deposition. All EISCAP systems have a clear pH response, favoring the stacked gate insulators SiO2–Ta2O5 when considering the overall sensor characteristics, while the Si3N4(1 nm)–Ta2O5 stack delivers the largest accumulation capacitance (due to the lower equivalent oxide thickness) and a higher steepness in the slope of the capacitance–voltage curve among the studied stacked gate insulator systems.
This study focuses on thermoelectric elements (TEE) as an alternative for room temperature control. TEE are semi-conductor devices that can provide heating and cooling via a heat pump effect without direct noise emissions and no refrigerant use. An efficiency evaluation of the optimal operating mode is carried out for different numbers of TEE, ambient temperatures, and heating loads. The influence of an additional heat recovery unit on system efficiency and an unevenly distributed heating demand are examined. The results show that TEE can provide heat at a coefficient of performance (COP) greater than one especially for small heating demands and high ambient temperatures. The efficiency increases with the number of elements in the system and is subject to economies of scale. The best COP exceeds six at optimal operating conditions. An additional heat recovery unit proves beneficial for low ambient temperatures and systems with few TEE. It makes COPs above one possible at ambient temperatures below 0 ∘C. The effect increases efficiency by maximal 0.81 (from 1.90 to 2.71) at ambient temperature 5 K below room temperature and heating demand Q˙h=100W but is subject to diseconomies of scale. Thermoelectric technology is a valuable option for electricity-based heat supply and can provide cooling and ventilation functions. A careful system design as well as an additional heat recovery unit significantly benefits the performance. This makes TEE superior to direct current heating systems and competitive to heat pumps for small scale applications with focus on avoiding noise and harmful refrigerants.
Resilience as a concept has found its way into different disciplines to describe the ability of an individual or system to withstand and adapt to changes in its environment. In this paper, we provide an overview of the concept in different communities and extend it to the area of mechanical engineering. Furthermore, we present metrics to measure resilience in technical systems and illustrate them by applying them to load-carrying structures. By giving application examples from the Collaborative Research Centre (CRC) 805, we show how the concept of resilience can be used to control uncertainty during different stages of product life.
Purpose Vascular risk factors and ocular perfusion are heatedly discussed in the pathogenesis of glaucoma. The retinal vessel analyzer (RVA, IMEDOS Systems, Germany) allows noninvasive measurement of retinal vessel regulation. Significant differences especially in the veins between healthy subjects and patients suffering from glaucoma were previously reported. In this pilot-study we investigated if localized vascular regulation is altered in glaucoma patients with altitudinal visual field defect asymmetry. Methods 15 eyes of 12 glaucoma patients with advanced altitudinal visual field defect asymmetry were included. The mean defect was calculated for each hemisphere separately (-20.99 ± 10.49 pro- found hemispheric visual field defect vs -7.36 ± 3.97 dB less profound hemisphere). After pupil dilation, RVA measurements of retinal arteries and veins were conducted using the standard protocol. The superior and inferior retinal vessel reactivity were measured consecutively in each eye. Results Significant differences were recorded in venous vessel constriction after flicker light stimulation and overall amplitude of the reaction (p \ 0.04 and p \ 0.02 respectively) in-between the hemispheres spheres. Vessel reaction was higher in the hemisphere corresponding to the more advanced visual field defect. Arterial diameters reacted similarly, failing to reach statistical significance. Conclusion Localized retinal vessel regulation is significantly altered in glaucoma patients with asymmetri altitudinal visual field defects. Veins supplying the hemisphere concordant to a less profound visual field defect show diminished diameter changes. Vascular dysregulation might be particularly important in early glaucoma stages prior to a significant visual field defect.
Given industrial applications, the costs for the operation and maintenance of a pump system typically far exceed its purchase price. For finding an optimal pump configuration which minimizes not only investment, but life-cycle costs, methods like Technical Operations Research which is based on Mixed-Integer Programming can be applied. However, during the planning phase, the designer is often faced with uncertain input data, e.g. future load demands can only be estimated. In this work, we deal with this uncertainty by developing a chance-constrained two-stage (CCTS) stochastic program. The design and operation of a booster station working under uncertain load demand are optimized to minimize total cost including purchase price, operation cost incurred by energy consumption and penalty cost resulting from water shortage. We find optimized system layouts using a sample average approximation (SAA) algorithm, and analyze the results for different risk levels of water shortage. By adjusting the risk level, the costs and performance range of the system can be balanced, and thus the
system’s resilience can be engineered
On obligations in the development process of resilient systems with algorithmic design methods
(2018)
Advanced computational methods are needed both for the design of large systems and to compute high accuracy solutions. Such methods are efficient in computation, but the validation of results is very complex, and highly skilled auditors are needed to verify them. We investigate legal questions concerning obligations in the development phase, especially for technical systems developed using advanced methods. In particular, we consider methods of resilient and robust optimization. With these techniques, high performance solutions can be found, despite a high variety of input parameters. However, given the novelty of these methods, it is uncertain whether legal obligations are being met. The aim of this paper is to discuss if and how the choice of a specific computational method affects the developer’s product liability. The review of legal obligations in this paper is based on German law and focuses on the requirements that must be met during the design and development process.
Algorithmic design and resilience assessment of energy efficient high-rise water supply systems
(2018)
High-rise water supply systems provide water flow and suitable pressure in all levels of tall buildings. To design such state-of-the-art systems, the consideration of energy efficiency and the anticipation of component failures are mandatory. In this paper, we use Mixed-Integer Nonlinear Programming to compute an optimal placement of pipes and pumps, as well as an optimal control strategy.Moreover, we consider the resilience of the system to pump failures. A resilient system is able to fulfill a predefined minimum functionality even though components fail or are restricted in their normal usage. We present models to measure and optimize the resilience. To demonstrate our approach, we design and analyze an optimal resilient decentralized water supply system inspired by a real-life hotel building.