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An amperometric bi-enzyme biosensor based on substrate recycling principle for the amplification of the sensor signal has been developed for the detection of adrenaline in blood. Adrenaline can be used as biomarker verifying successful adrenal venous sampling procedure. The adrenaline biosensor has been realized via modification of a galvanic oxygen sensor with a bi-enzyme membrane combining a genetically modified laccase and a pyrroloquinoline quinone-dependent glucose dehydrogenase. The measurement conditions such as pH value and temperature were optimized to enhance the sensor performance. A high sensitivity and a low detection limit of about 0.5–1 nM adrenaline have been achieved in phosphate buffer at pH 7.4, relevant for measurements in blood samples. The sensitivity of the biosensor to other catecholamines such as noradrenaline, dopamine and dobutamine has been studied. Finally, the sensor has been successfully applied for the detection of adrenaline in human blood plasma.
Field-effect-based electrolyte-insulator-semiconductor (EIS) sensors were modified with a bilayer of positively charged weak polyelectrolyte (poly(allylamine hydrochloride) (PAH)) and probe single-stranded DNA (ssDNA) and are used for the detection of complementary single-stranded target DNA (cDNA) in different test solutions. The sensing mechanism is based on the detection of the intrinsic molecular charge of target cDNA molecules after the hybridization event between cDNA and immobilized probe ssDNA. The test solutions contain synthetic cDNA oligonucleotides (with a sequence of tuberculosis mycobacteria genome) or PCR-amplified DNA (which origins from a template DNA strand that has been extracted from Mycobacterium avium paratuberculosis-spiked human sputum samples), respectively. Sensor responses up to 41 mV have been measured for the test solutions with DNA, while only small signals of ∼5 mV were detected for solutions without DNA. The lower detection limit of the EIS sensors was ∼0.3 nM, and the sensitivity was ∼7.2 mV/decade. Fluorescence experiments using SybrGreen I fluorescence dye support the electrochemical results.
A capacitive electrolyte-insulator-semiconductor (EIS) field-effect biosensor for acetoin detection has been presented for the first time. The EIS sensor consists of a layer structure of Al/p-Si/SiO₂/Ta₂O₅/enzyme acetoin reductase. The enzyme, also referred to as butane-2,3-diol dehydrogenase from B. clausii DSM 8716T, has been recently characterized. The enzyme catalyzes the (R)-specific reduction of racemic acetoin to (R,R)- and meso-butane-2,3-diol, respectively. Two different enzyme immobilization strategies (cross-linking by using glutaraldehyde and adsorption) have been studied. Typical biosensor parameters such as optimal pH working range, sensitivity, hysteresis, linear concentration range and long-term stability have been examined by means of constant-capacitance (ConCap) mode measurements. Furthermore, preliminary experiments have been successfully carried out for the detection of acetoin in diluted white wine samples.
Cupriavidus necator H16 gains increasing attention in microbial research and biotechnological application due to its diverse metabolic features. Here we present a tightly controlled gene expression system for C. necator including the pBBR1-vector that contains hybrid promoters originating from C. necator native tolC-promoter in combination with a synthetic tetO-operator. The expression of the reporter gene from these plasmids relies on the addition of the exogenous inducer doxycycline (dc). The novel expression system offers a combination of advantageous features as; (i) high and dose-dependent recombinant protein production, (ii) tight control with a high dynamic range (On/Off ratio), which makes it applicable for harmful pathways or for toxic protein production, (iii) comparable cheap inducer (doxycycline, dc), (iv) effective at low inducer concentration, that makes it useful for large scale application, (v) rapid, diffusion controlled induction, and (vi) the inducer does not interfere within the cell metabolism. As applications of the expression system in C. necator H16, the growth ability on glycerol was enhanced by constitutively expressing the E. coli glpk gene-encoding for glycerol kinase. Likewise, we used the system to overcome the expression toxicity of mevalonate pathway in C. necator H16. With this system, the mevalonate-genes were successfully introduced in the host and the recombinant strains could produce about 200 mg/l mevalonate.
In this paper, a coupled multiphase model considering both non-linearities of water retention curves and solid state modeling is proposed. The solid displacements and the pressures of both water and air phases are unknowns of the proposed model. The finite element method is used to solve the governing differential equations. The proposed method is demonstrated through simulation of seepage test and partially consolidation problem. Then, implementation of the model is done by using hypoplasticity for the solid phase and analyzing the fully saturated triaxial experiments. In integration of the constitutive law error controlling is improved and comparisons done accordingly. In this work, the advantages and limitations of the numerical model are discussed.
Durch die Bauforschung im Rahmen des mehrjährigen Projektes "Die Hochkorridore von Sabbioneta" konnten der Verlauf und viele Anschlüsse der hochliegenden Gänge an bestehende Bauten weitgehend geklärt werden. Im Bereich der Chiesa della SS. Maria Incoronata und des angrenzenden Servitenklosters führten die Bauuntersuchungen zu völlig neuen Erkenntnissen über den ehemaligen Verlauf des Hochkorridors und damit zu einer Neuinterpretation des baulichen Ensembles sowie seiner Bedeutung für die Idealstadt. Aus dieser historischen Umdeutung entstanden Ideen für die Umnutzung des ehemaligen Servitenklosters und seiner angrenzenden Freiflächen, die im Rahmen von studentischen Entwürfen am Lehrstuhl für Baugeschichte und Denkmalpflege in Kooperation mit dem Lehrstuhl für Landschaftsarchitektur der RWTH Aachen weiterentwickelt wurden. Somit stellten sich Befunde einer historisch orientierten Bauforschung mit übergeordneter Fragestellung als neue Impulsgeber für Stadtplanung, Architektur und Denkmalpflege heraus.
During rapid deceleration of the body, tendons buffer part of the elongation of the muscle-tendon unit (MTU), enabling safe energy dissipation via eccentric muscle contraction. Yet, the influence of changes in tendon stiffness within the physiological range upon these lengthening contractions is unknown. This study aimed to examine the effect of training-induced stiffening of the Achilles tendon on triceps surae muscle-tendon behavior during a landing task. Twenty-one male subjects were assigned to either a 10-week resistance-training program consisting of single-leg isometric plantarflexion (n = 11) or to a non-training control group (n = 10). Before and after the training period, plantarflexion force, peak Achilles tendon strain and stiffness were measured during isometric contractions, using a combination of dynamometry, ultrasound and kinematics data. Additionally, testing included a step-landing task, during which joint mechanics and lengths of gastrocnemius and soleus fascicles, Achilles tendon, and MTU were determined using synchronized ultrasound, kinematics and kinetics data collection. After training, plantarflexion strength and Achilles tendon stiffness increased (15 and 18%, respectively), and tendon strain during landing remained similar. Likewise, lengthening and negative work produced by the gastrocnemius MTU did not change detectably. However, in the training group, gastrocnemius fascicle length was offset (8%) to a longer length at touch down and, surprisingly, fascicle lengthening and velocity were reduced by 27 and 21%, respectively. These changes were not observed for soleus fascicles when accounting for variation in task execution between tests. These results indicate that a training-induced increase in tendon stiffness does not noticeably affect the buffering action of the tendon when the MTU is rapidly stretched. Reductions in gastrocnemius fascicle lengthening and lengthening velocity during landing occurred independently from tendon strain. Future studies are required to provide insight into the mechanisms underpinning these observations and their influence on energy dissipation.
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.
Das anhaltende Wachstum wissenschaftlicher Veröffentlichungen wirft die Fragestellung auf, wie Literaturana-lysen im Rahmen von Forschungsprozessen digitalisiert und somit produktiver realisiert werden können. Insbesondere in informationstechnischen Fachgebieten ist die Forschungspraxis durch ein rasant wachsendes Publikationsaufkommen gekennzeichnet. Infolgedessen bietet sich der Einsatz von Methoden der Textanalyse (Text Analytics) an, die Textdaten automatisch vorbereiten und verarbeiten können. Erkenntnisse entstehen dabei aus Analysen von Wortarten und Subgruppen, Korrelations- sowie Zeitreihenanalysen. Dieser Beitrag stellt die Konzeption und Realisierung eines Prototypen vor, mit dem Anwender bibliographische Daten aus der etablierten Literaturdatenbank EBSCO Discovery Service mithilfe textanalytischer Methoden erschließen können. Der Prototyp basiert auf dem Analysesystem IBM Watson Explorer, das Hochschulen lizenzkostenfrei zur Verfügung steht. Potenzielle Adressaten des Prototypen sind Forschungseinrichtungen, Beratungsunternehmen sowie Entscheidungsträger in Politik und Unternehmenspraxis.
In lab-on-chip systems, electrodes are important for the manipulation (e.g., cell stimulation, electrolysis) within such systems. An alternative to commonly used electrode structures can be a light-addressable electrode. Here, due to the photoelectric effect, the conducting area can be adjusted by modification of the illumination area which enables a flexible control of the electrode. In this work, titanium dioxide based light-addressable electrodes are fabricated by a sol–gel technique and a spin-coating process, to deposit a thin film on a fluorine-doped tin oxide glass. To characterize the fabricated electrodes, the thickness, and morphological structure are measured by a profilometer and a scanning electron microscope. For the electrochemical behavior, the dark current and the photocurrent are determined for various film thicknesses. For the spatial resolution behavior, the dependency of the photocurrent while changing the area of the illuminated area is studied. Furthermore, the addressing of single fluid compartments in a three-chamber system, which is added to the electrode, is demonstrated.
Background
Culture media containing complex compounds like yeast extract or peptone show numerous disadvantages. The chemical composition of the complex compounds is prone to significant variations from batch to batch and quality control is difficult. Therefore, the use of chemically defined media receives more and more attention in commercial fermentations. This concept results in better reproducibility, it simplifies downstream processing of secreted products and enable rapid scale-up. Culturing bacteria with unknown auxotrophies in chemically defined media is challenging and often not possible without an extensive trial-and-error approach. In this study, a respiration activity monitoring system for shake flasks and its recent version for microtiter plates were used to clarify unknown auxotrophic deficiencies in the model organism Bacillus pumilus DSM 18097.
Results
Bacillus pumilus DSM 18097 was unable to grow in a mineral medium without the addition of complex compounds. Therefore, a rich chemically defined minimal medium was tested containing basically all vitamins, amino acids and nucleobases, which are essential ingredients of complex components. The strain was successfully cultivated in this medium. By monitoring of the respiration activity, nutrients were supplemented to and omitted from the rich chemically defined medium in a rational way, thus enabling a systematic and fast determination of the auxotrophic deficiencies. Experiments have shown that the investigated strain requires amino acids, especially cysteine or histidine and the vitamin biotin for growth.
Conclusions
The introduced method allows an efficient and rapid identification of unknown auxotrophic deficiencies and can be used to develop a simple chemically defined tailor-made medium. B. pumilus DSM 18097 was chosen as a model organism to demonstrate the method. However, the method is generally suitable for a wide range of microorganisms. By combining a systematic combinatorial approach based on monitoring the respiration activity with cultivation in microtiter plates, high throughput experiments with high information content can be conducted. This approach facilitates media development, strain characterization and cultivation of fastidious microorganisms in chemically defined minimal media while simultaneously reducing the experimental effort.
New information regarding the influence of a stepped chute on the hydraulic performance of the United States Bureau of Reclamation (Reclamation) Type III hydraulic jump stilling basin is presented for design (steady) and adverse (decreasing tailwater) conditions. Using published experimental data and computational fluid dynamics (CFD) models, this paper presents a detailed comparison between smooth-chute and stepped-chute configurations for chute slopes of 0.8H:1V and 4H:1V and Froude numbers (F) ranging from 3.1 to 9.5 for a Type III basin designed for F = 8. For both stepped and smooth chutes, the relative role of each basin element was quantified, up to the most hydraulic extreme case of jump sweep-out. It was found that, relative to a smooth chute, the turbulence generated by a stepped chute causes a higher maximum velocity decay within the stilling basin, which represents an enhancement of the Type III basin’s performance but also a change in the relative role of the basin elements. Results provide insight into the ability of the CFD models [unsteady Reynolds-averaged Navier-Stokes (RANS) equations with renormalization group (RNG) k-ϵ turbulence model and volume-of-fluid (VOF) for free surface tracking] to predict the transient basin flow structure and velocity profiles. Type III basins can perform adequately with a stepped chute despite the effects steps have on the relative role of each basin element. It is concluded that the classic Type III basin design, based upon methodology by reclamation specific to smooth chutes, can be hydraulically improved for the case of stepped chutes for design and adverse flow conditions using the information presented herein.
For fuel flexibility enhancement hydrogen represents a possible alternative gas turbine fuel within future low emission power generation, in case of hydrogen production by the use of renewable energy sources such as wind energy or biomass. Kawasaki Heavy Industries, Ltd. (KHI) has research and development projects for future hydrogen society; production of hydrogen gas, refinement and liquefaction for transportation and storage, and utilization with gas turbine / gas engine for the generation of electricity. In the development of hydrogen gas turbines, a key technology is the stable and low NOx hydrogen combustion, especially Dry Low Emission (DLE) or Dry Low NOx (DLN) hydrogen combustion. Due to the large difference in the physical properties of hydrogen compared to other fuels such as natural gas, well established gas turbine combustion systems cannot be directly applied for DLE hydrogen combustion. Thus, the development of DLE hydrogen combustion technologies is an essential and challenging task for the future of hydrogen fueled gas turbines. The DLE Micro-Mix combustion principle for hydrogen fuel has been in development for many years to significantly reduce NOx emissions. This combustion principle is based on cross-flow mixing of air and gaseous hydrogen which reacts in multiple miniaturized “diffusion-type” flames. The major advantages of this combustion principle are the inherent safety against flashback and the low NOx-emissions due to a very short residence time of the reactants in the flame region of the micro-flames.
Stahlbetonrahmentragwerke mit Mauerwerksausfachungen weisen nach Erdbebenereignissen häufig schwere Schäden auf, da die Ausfachungen ohne weitere konstruktive Maßnahmen mit vollem Kontakt zum Stahlbetonrahmen eingemauert werden. Durch die unplanmäßige Beteiligung am horizontalen Lastabtrag erfahren die Ausfachungen Belastungen in Wandebene und beeinflussen das globale Schwingungsverhalten der Rahmentragwerke. In Kombination mit den gleichzeitig auftretenden seismischen Trägheitskräften senkrecht zur Wand führt dies in vielen Fällen zu einem Versagen der mit niedrigen Festigkeiten ausgeführten Ausfachungen. Dies war der Anlass in dem europäischen Forschungsprojekt INSYSME ein Entkopplungssystem zu entwickeln, mit dem Rahmen und Ausfachung durch ein spezielles Profil aus Elastomeren entkoppelt werden.
Das Profil ermöglicht Relativverschiebungen zwischen Rahmen und Ausfachung und stellt gleichzeitig die Aufnahme von Belastungen senkrecht zur Wand sicher. Der Beitrag erläutert zunächst den Aufbau des Systems und gibt einen Überblick über die in Kleinbauteilversuchen ermittelten Tragfähigkeiten. Zudem werden experimentelle Untersuchungen an mit hochwärmedämmenden Mauerziegeln ausgefachten Stahlbetonrahmen mit und ohne Entkopplungssystem für getrennte und kombinierte Belastungen in und senkrecht zur Wandebene vorgestellt. Auf Grundlage einer Versuchsauswertung und eines Ergebnisvergleichs werden Wirkungsweise und Effektivität des entwickelten Entkopplungssystems demonstriert.
In this work, we report on our attempt to design and implement an early introduction to basic robotics principles for children at kindergarten age. One of the main challenges of this effort is to explain complex robotics contents in a way that pre-school children could follow the basic principles and ideas using examples from their world of experience. What sets apart our effort from other work is that part of the lecturing is actually done by a robot itself and that a quiz at the end of the lesson is done using robots as well. The humanoid robot Pepper from Softbank, which is a great platform for human–robot interaction experiments, was used to present a lecture on robotics by reading out the contents to the children making use of its speech synthesis capability. A quiz in a Runaround-game-show style after the lecture activated the children to recap the contents they acquired about how mobile robots work in principle. In this quiz, two LEGO Mindstorm EV3 robots were used to implement a strongly interactive scenario. Besides the thrill of being exposed to a mobile robot that would also react to the children, they were very excited and at the same time very concentrated. We got very positive feedback from the children as well as from their educators. To the best of our knowledge, this is one of only few attempts to use a robot like Pepper not as a tele-teaching tool, but as the teacher itself in order to engage pre-school children with complex robotics contents.
BACKGROUND: Muscle stretch reflexes are widely considered to beneficially influence joint stability and power generation in the lower limbs. While in the upper limbs and especially in the muscles surrounding the shoulder joint such evidence is lacking. OBJECTIVE: To quantify the electromyographical response in the muscles crossing the shoulder of specifically trained overhead athletes to an anterior perturbation force. METHODS: Twenty healthy male participants performed six sets of different external shoulder rotation stretches on an isokinetic dynamometer over a range of amplitudes and muscle pre-activation moment levels. All stretches were applied with a dynamometer acceleration of 10,000∘/s2 and a velocity of 150∘/s. Electromyographical response was measured via sEMG. RESULTS: Consistent reflexes were not observed in all experimental conditions. The reflex latencies revealed a significant muscle main effect (F (2,228) = 99.31, p< 0.001; η2= 0.466; f= 0.934) and a pre-activation main effect (F (1,228) = 142.21, p< 0.001; η2= 0.384; f= 1.418). The stretch reflex amplitude yielded a significant pre-activation main effect (F (1,222) = 470.373, p< 0.001; η2= 0.679; f= 1.454). CONCLUSION: Short latency muscle reflexes showed a tendency to an anterior to posterior muscle recruitment whereby the main internal rotator muscles of the shoulder revealed the most consistent results.