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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.
Ambitious climate targets affect the competitiveness of industries in the international market. To prevent such industries from moving to other countries in the wake of increased climate protection efforts, cost adjustments may become necessary. Their design requires knowledge of country-specific production costs. Here, we present country-specific cost figures for different production routes of steel, paying particular attention to transportation costs. The data can be used in floor price models aiming to assess the competitiveness of different steel production routes in different countries (Rübbelke, 2022).
Penner, Crystal ; Usherovich, Samuel ; Niedermeier, Jana ; Bélanger-Champagne, Camille ; Trinczek, Michael ; Paulßen, Elisabeth ; Hoehr, Cornelia
In proton therapy, the dose from secondary neutrons to the patient can contribute to side effects and the creation of secondary cancer. A simple and fast detection system to distinguish between dose from protons and neutrons both in pretreatment verification as well as potentially in vivo monitoring is needed to minimize dose from secondary neutrons. Two 3 mm long, 1 mm diameter organic scintillators were tested for candidacy to be used in a proton–neutron discrimination detector. The SCSF-3HF (1500) scintillating fibre (Kuraray Co. Chiyoda-ku, Tokyo, Japan) and EJ-260 plastic scintillator (Eljen Technology, Sweetwater, TX, USA) were irradiated at the TRIUMF Neutron Facility and the Proton Therapy Research Centre. In the proton beam, we compared the raw Bragg peak and spread-out Bragg peak response to the industry standard Markus chamber detector. Both scintillator sensors exhibited quenching at high LET in the Bragg peak, presenting a peak-to-entrance ratio of 2.59 for the EJ-260 and 2.63 for the SCSF-3HF fibre, compared to 3.70 for the Markus chamber. The SCSF-3HF sensor demonstrated 1.3 times the sensitivity to protons and 3 times the sensitivity to neutrons as compared to the EJ-260 sensor. Combined with our equations relating neutron and proton contributions to dose during proton irradiations, and the application of Birks’ quenching correction, these fibres provide valid candidates for inexpensive and replicable proton-neutron discrimination detectors
Optical Fibers as Dosimeter Detectors for Mixed Proton/Neutron Fields - A Biological Dosimeter
(2023)
Niedermeier, Jana ; Penner, Crystal ; Usherovich, Samuel ; Bélanger-Champagne, Camille ; Paulßen, Elisabeth ; Hoehr, Cornelia
In recent years, proton therapy has gained importance as a cancer treatment modality due to its conformality with the tumor and the sparing of healthy tissue. However, in the interaction of the protons with the beam line elements and patient tissues, potentially harmful secondary neutrons are always generated. To ensure that this neutron dose is as low as possible, treatment plans could be created to also account for and minimize the neutron dose. To monitor such a treatment plan, a compact, easy to use, and inexpensive dosimeter must be developed that not only measures the physical dose, but which can also distinguish between proton and neutron contributions. To that end, plastic optical fibers with scintillation materials (Gd₂O₂S:Tb, Gd₂O₂S:Eu, and YVO₄:Eu) were irradiated with protons and neutrons. It was confirmed that sensors with different scintillation materials have different sensitivities to protons and neutrons. A combination of these three scintillators can be used to build a detector array to create a biological dosimeter.
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.
Aufgrund der gestiegenen Anforderungen durch höhere Ein-wirkungen aus Wind und Erdbeben ist eine Verbesserung und Optimierung der Berechnungs- und Bemessungsansätze für Mauerwerksbauten erforderlich. Eine bessere Ausnutzung der Tragwerksreserven ist durch die Berücksichtigung der Rah-mentragwirkung mit einer Aktivierung der Deckenscheiben in den Rechenmodellen möglich, die in der Praxis aufgrund der Komplexität der Wand-Decken-Interaktion bislang nicht aus-genutzt wird. Im vorliegenden Aufsatz wird ein vereinfachter Ansatz auf Grundlage der mitwirkenden Plattenbreite von Schubwänden aus Mauerwerk vorgestellt, der die wesentli-chen Einfl ussfaktoren in parametrisierten Tabellen erfasst. Damit steht den Tragwerksplanern ein einfach anwendbares Werkzeug zur Verfügung, um die Rahmentragwirkung in der Mauerwerksbemessung anzusetzen.
Im Norden von Serbien erfolgt in Novi Sad der Neubau eines modernen Forschungsgebäudes für das BioSense-Institut mit finanzieller Unterstützung durch die Eu-ropäische Union. Der Gebäudeteil mit Laboren wird zum Schutz und zur Sicherstellung des reibungslosen Betriebs der sensiblen und kapitalintensiven technischen Einbauten mit ei-ner Erdbebenisolierung mit integrierter Körperschallisolation versehen. Zusätzlich wird der entkoppelte Laborteil des For-schungsgebäudes mit einem BIM-basierten Bauwerksmonito-ring versehen, um Änderungen des Gebäudezustands jederzeit abfragen und beurteilen zu können.
Recent earthquakes as the 2012 Emilia earthquake sequence showed that recently built unreinforced masonry (URM) buildings behaved much better than expected and sustained, despite the maximum PGA values ranged between 0.20–0.30 g, either minor damage or structural damage that is deemed repairable. Especially low-rise residential and commercial masonry buildings with a code-conforming seismic design and detailing behaved in general very well without substantial damages. The low damage grades of modern masonry buildings that was observed during this earthquake series highlighted again that codified design procedures based on linear analysis can be rather conservative. Although advances in simulation tools make nonlinear calculation methods more readily accessible to designers, linear analyses will still be the standard design method for years to come. The present paper aims to improve the linear seismic design method by providing a proper definition of the q-factor of URM buildings. These q-factors are derived for low-rise URM buildings with rigid diaphragms which represent recent construction practise in low to moderate seismic areas of Italy and Germany. The behaviour factor components for deformation and energy dissipation capacity and for overstrength due to the redistribution of forces are derived by means of pushover analyses. Furthermore, considerations on the behaviour factor component due to other sources of overstrength in masonry buildings are presented. As a result of the investigations, rationally based values of the behaviour factor q to be used in linear analyses in the range of 2.0–3.0 are proposed.
Im Handel mit Kraftfahrzeugen gehören Aspekte des gutgläubigen Erwerbs zu den beinahe alltäglichen Standardproblemen. Der BGH fügt in seiner Entscheidung v. 23.9.2022–VZR148/21, MDR 2022, 1541 diesem im Detail breit gefächerten Themenfeld einen weiteren Mosaikstein hinzu: Der Erwerber erhielt das verkaufte Kfz ohne Übergabe einer Zulassungsbescheinigung Teil II, behauptet aber, diese Bescheinigung sei dem vom ihm eingeschalteten Vermittler bei Erwerb (als Fälschung) vorgelegt worden. Tatsächlich befand sich das Original durchgängig beim wahren Eigentümer, der nunmehr Herausgabe des Fahrzeugs verlangt. Der BGH schützt in dieser Gestaltung im Ergebnis den Erwerber. Die Entscheidung ist in mehrfacher Hinsicht bemerkenswert.
Because of simple construction process, high energy efficiency, significant fire resistance and excellent sound isolation, masonry infilled reinforced concrete (RC) frame structures are very popular in most of the countries in the world, as well as in seismic active areas. However, many RC frame structures with masonry infills were seriously damaged during earthquake events, as the traditional infills are generally constructed with direct contact to the RC frame which brings undesirable infill/frame interaction. This interaction leads to the activation of the equivalent diagonal strut in the infill panel, due to the RC frame deformation, and combined with seismically induced loads perpendicular to the infill panel often causes total collapses of the masonry infills and heavy damages to the RC frames. This fact was the motivation for developing different approaches for improving the behaviour of masonry infills, where infill isolation (decoupling) from the frame has been more intensively studied in the last decade. In-plane isolation of the infill wall reduces infill activation, but causes the need for additional measures to restrain out-of-plane movements. This can be provided by installing steel anchors, as proposed by some researchers. Within the framework of European research project INSYSME (Innovative Systems for Earthquake Resistant Masonry Enclosures in Reinforced Concrete Buildings) the system based on a use of elastomers for in-plane decoupling and steel anchors for out-of-plane restrain was tested. This constructive solution was tested and deeply investigated during the experimental campaign where traditional and decoupled masonry infilled RC frames with anchors were subjected to separate and combined in-plane and out-of-plane loading. Based on a detailed evaluation and comparison of the test results, the performance and effectiveness of the developed system are illustrated.
Erdbebennachweis von Mauerwerksbauten mit realistischen Modellen und erhöhten Verhaltensbeiwerten
(2021)
Die Anwendung des linearen Nachweiskonzepts auf Mauerwerksbauten führt dazu, dass bereits heute Standsicherheitsnachweise für Gebäude mit üblichen Grundrissen in Gebieten mit moderaten Erdbebeneinwirkungen nicht mehr geführt werden können. Diese Problematik wird sich in Deutschland mit der Einführung kontinuierlicher probabilistischer Erdbebenkarten weiter verschärfen. Aufgrund der Erhöhung der seismischen Einwirkungen, die sich vielerorts ergibt, ist es erforderlich, die vorhandenen, bislang nicht berücksichtigten Tragfähigkeitsreserven in nachvollziehbaren Nachweiskonzepten in der Baupraxis verfügbar zu machen. Der vorliegende Beitrag stellt ein Konzept für die gebäudespezifische Ermittlung von erhöhten Verhaltensbeiwerten vor. Die Verhaltensbeiwerte setzen sich aus drei Anteilen zusammen, mit denen die Lastumverteilung im Grundriss, die Verformungsfähigkeit und Energiedissipation sowie die Überfestigkeiten berücksichtigt werden. Für die rechnerische Ermittlung dieser drei Anteile wird ein nichtlineares Nachweiskonzept auf Grundlage von Pushover-Analysen vorgeschlagen, in denen die Interaktionen von Wänden und Geschossdecken durch einen Einspanngrad beschrieben werden. Für die Bestimmung der Einspanngrade wird ein nichtlinearer Modellierungsansatz eingeführt, mit dem die Interaktion von Wänden und Decken abgebildet werden kann. Die Anwendung des Konzepts mit erhöhten gebäudespezifischen Verhaltensbeiwerten wird am Beispiel eines Mehrfamilienhauses aus Kalksandsteinen demonstriert. Die Ergebnisse der linearen Nachweise mit erhöhten Verhaltensbeiwerten für dieses Gebäude liegen deutlich näher an den Ergebnissen nichtlinearer Nachweise und somit bleiben übliche Grundrisse in Erdbebengebieten mit den traditionellen linearen Rechenansätzen nachweisbar.
Monte Carlo Tree Search (MCTS) is a search technique that in the last decade emerged as a major breakthrough for Artificial Intelligence applications regarding board- and video-games. In 2016, AlphaGo, an MCTS-based software agent, outperformed the human world champion of the board game Go. This game was for long considered almost infeasible for machines, due to its immense search space and the need for a long-term strategy. Since this historical success, MCTS is considered as an effective new approach for many other scientific and technical problems. Interestingly, civil structural engineering, as a discipline, offers many tasks whose solution may benefit from intelligent search and in particular from adopting MCTS as a search tool. In this work, we show how MCTS can be adapted to search for suitable solutions of a structural engineering design problem. The problem consists of choosing the load-bearing elements in a reference reinforced concrete structure, so to achieve a set of specific dynamic characteristics. In the paper, we report the results obtained by applying both a plain and a hybrid version of single-agent MCTS. The hybrid approach consists of an integration of both MCTS and classic Genetic Algorithm (GA), the latter also serving as a term of comparison for the results. The study’s outcomes may open new perspectives for the adoption of MCTS as a design tool for civil engineers.
Mauerwerksbauten in Deutschland sind mit Einführung des nationalen Anwendungsdokuments DIN EN 1998-1/NA auf Grundlage einer neuen probabilistischen Erdbebenkarte nachzuweisen. Für erfolgreiche Erdbebennachweise üblicher Grundrissformen von Mauerwerksbauten stehen in dem zukünftigen Anwendungsdokument neue rechnerische Nachweismöglichkeiten zur Verfügung, mit denen die Tragfähigkeitsreserven von Mauerwerksbauten in der Baupraxis mit einem überschaubaren Aufwand besser in Ansatz gebracht werden können. Das Standardrechenverfahren ist weiterhin der kraftbasierte Nachweis, der nun mit höheren Verhaltensbeiwerten im Vergleich zur DIN 4149 durchgeführt werden kann. Die höheren Verhaltensbeiwerte basieren auf der besseren Ausnutzung der gebäudespezifischen Verformungsfähigkeit und Energiedissipation sowie der Lastumverteilung der Schubkräfte im Grundriss mit Ansatz von Rahmentragwirkung durch Wand-Deckeninteraktionen. Alternativ dazu kann ein nichtlinearer Nachweis auf Grundlage von Pushover-Analysen zur Anwendung kommen. Vervollständigt werden die Regelungen für Mauerwerksbauten durch neue Regelungen für nichttragende Innenwände und Außenmauerschalen. Der vorliegende Beitrag stellt die Grundlagen und Hintergründe der neuen rechnerischen Nachweise in DIN EN 1998-1/NA vor und demonstriert deren Anwendung an einem Beispiel aus der Praxis.
Butenweg, Christoph ; Bursi, Oreste S. ; Paolacci, Fabrizio ; Marinković, Marko ; Lanese, Igor ; Nardin, Chiara ; Quinci, Gianluca
Past earthquakes demonstrated the high vulnerability of industrial facilities equipped with complex process technologies leading to serious damage of process equipment and multiple and simultaneous release of hazardous substances. Nonetheless, current standards for seismic design of industrial facilities are considered inadequate to guarantee proper safety conditions against exceptional events entailing loss of containment and related consequences. On these premises, the SPIF project -Seismic Performance of Multi-Component Systems in Special Risk Industrial Facilities- was proposed within the framework of the European H2020 SERA funding scheme. In detail, the objective of the SPIF project is the investigation of the seismic behaviour of a representative industrial multi-storey frame structure equipped with complex process components by means of shaking table tests. Along this main vein and in a performance-based design perspective, the issues investigated in depth are the interaction between a primary moment resisting frame (MRF) steel structure and secondary process components that influence the performance of the whole system; and a proper check of floor spectra predictions. The evaluation of experimental data clearly shows a favourable performance of the MRF structure, some weaknesses of local details due to the interaction between floor crossbeams and process components and, finally, the overconservatism of current design standards w.r.t. floor spectra predictions.
In a special paired sample case, Hotelling’s T² test based on the differences of the paired random vectors is the likelihood ratio test for testing the hypothesis that the paired random vectors have the same mean; with respect to a special group of affine linear transformations it is the uniformly most powerful invariant test for the general alternative of a difference in mean. We present an elementary straightforward proof of this result. The likelihood ratio test for testing the hypothesis that the covariance structure is of the assumed special form is derived and discussed. Applications to real data are given.
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.
The paper deals with the asymptotic behaviour of estimators, statistical tests and confidence intervals for L²-distances to uniformity based on the empirical distribution function, the integrated empirical distribution function and the integrated empirical survival function. Approximations of power functions, confidence intervals for the L²-distances and statistical neighbourhood-of-uniformity validation tests are obtained as main applications. The finite sample behaviour of the procedures is illustrated by a simulation study.
Suppose we have k samples X₁,₁,…,X₁,ₙ₁,…,Xₖ,₁,…,Xₖ,ₙₖ with different sample sizes ₙ₁,…,ₙₖ and unknown underlying distribution functions F₁,…,Fₖ as observations plus k families of distribution functions {G₁(⋅,ϑ);ϑ∈Θ},…,{Gₖ(⋅,ϑ);ϑ∈Θ}, each indexed by elements ϑ from the same parameter set Θ, we consider the new goodness-of-fit problem whether or not (F₁,…,Fₖ) belongs to the parametric family {(G₁(⋅,ϑ),…,Gₖ(⋅,ϑ));ϑ∈Θ}. New test statistics are presented and a parametric bootstrap procedure for the approximation of the unknown null distributions is discussed. Under regularity assumptions, it is proved that the approximation works asymptotically, and the limiting distributions of the test statistics in the null hypothesis case are determined. Simulation studies investigate the quality of the new approach for small and moderate sample sizes. Applications to real-data sets illustrate how the idea can be used for verifying model assumptions.
A nonparametric goodness-of-fit test for random variables with values in a separable Hilbert space is investigated. To verify the null hypothesis that the data come from a specific distribution, an integral type test based on a Cramér-von-Mises statistic is suggested. The convergence in distribution of the test statistic under the null hypothesis is proved and the test's consistency is concluded. Moreover, properties under local alternatives are discussed. Applications are given for data of huge but finite dimension and for functional data in infinite dimensional spaces. A general approach enables the treatment of incomplete data. In simulation studies the test competes with alternative proposals.
The efficiency concepts of Bahadur and Pitman are used to compare the Wilcoxon tests in paired and independent survey samples. A comparison through the length of corresponding confidence intervals is also done. Simple conditions characterizing the dominance of a procedure are derived. Statistical tests for checking these conditions are suggested and discussed.