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The use of highly expressed FTH1 as carrier protein for cytosolic targeting in Hansenula polymorpha
(2012)
The iron storage protein ferritin is a member of the non-heme iron protein family. It can store and release iron, therefore it prevents the cell from damage caused by iron-dioxygen reactions as well as it provides iron for biological processing. To study whether the human ferritin heavy chain (FTH1) can be expressed in Hansenula polymorpha, we integrated an expression cassette for FTH1 and analyzed the protein expression. We found very efficient expression of FTH1 and obtained yields up to 1.9 g/L under non-optimized conditions. Based on this result we designed a FTH1-PTH fusion protein to successfully express the parathyroid hormone fragment 1-34 (PTH) for the first time intracellular in H. polymorpha.
The literature as well as databases are ambiguous about the exact start of human interleukin-6 (IL-6)--three possibilities for the initiation of the mature protein are described. These three variants of IL-6, different in the exact initiation of the mature protein (A28, P29, or V30), were expressed in Hansenula polymorpha using the Saccharomyces cerevisiae MFα prepro sequence instead of the homologous pre sequence. All three IL-6 variants were secreted but the processing by the Kex2 protease showed significant differences. V30-IL-6 showed correctly processed material but also a molecule species of higher molecular weight indicating incomplete processing of the MFα pro peptide. P29-IL-6 did not yield any correctly processed IL-6, instead only the unprocessed pro form was found in the culture supernatant. Only A28-IL-6 led to 100% correctly processed material. N-terminal sequencing of this material revealed a start at V30--obviously the first two amino acids (Ala28-Pro29) have been removed by a so far unknown protease. Thus expression of both A28-IL-6 and V30-IL-6 as MFα prepro fusion proteins resulted in the very same mature V30-IL-6, however, the ratio of correctly processed molecules was significantly higher in the case of A28-IL-6. The expression of an MFα prepro-interferon α-2a (IFNα-2a) fusion protein in H. polymorpha leads to about 50% correctly processed molecules and 50% misprocessed forms which contain part of the pro peptide at the N-termini. The insertion of A28 and P29 of IL-6 between the pro peptide and the start of the mature IFNα-2a led to correct processing and elimination of all high molecular weight isoforms observed in earlier experiments.
Music listening has become a highly individualized activity with smartphones and music streaming services providing listeners with absolute freedom to listen to any kind of music in any situation. Until now, little has been written about the processes underlying the selection of music in daily life. The present study aimed to disentangle some of the complex processes among the listener, situation, and functions of music listening involved in music selection. Utilizing the experience sampling method, data were collected from 119 participants using a smartphone application. For 10 consecutive days, participants received 14 prompts using stratified-random sampling throughout the day and reported on their music-listening behavior. Statistical learning procedures on multilevel regression models and multilevel structural equation modeling were used to determine the most important predictors and analyze mediation processes between person, situation, functions of listening, and music selection. Results revealed that the features of music selected in daily life were predominantly determined by situational characteristics, whereas consistent individual differences were of minor importance. Functions of music listening were found to act as a mediator between characteristics of the situation and music-selection behavior. We further observed several significant random effects, which indicated that individuals differed in how situational variables affected their music selection behavior. Our findings suggest a need to shift the focus of music-listening research from individual differences to situational influences, including potential person-situation interactions.
Singing in different rooms: Common or individual adaptation patterns to the acoustic conditions?
(2020)
Das Agile Manifest (https://agilemanifesto.org) hat weltweit große Bedeutung erlangt. Der Begriff der Agilität ist allgegenwärtig, sei es in der Softwareentwicklung, im Projektmanagement oder in der Welt der Startups. Aber wie sieht es mit Bildungsprozessen aus? Möchte man nicht auch hier schnell und flexibel auf veränderte Anforderungen oder spontan entstehende Lernbedarfe reagieren? Und erlangen nicht auch hier kooperative, ergebnisoffenere und vor allem selbstorganisierte Lernprozesse zunehmend an Bedeutung, weil sie die Lernenden möglicherweise besser auf die zunehmend digitalisierte Lebens und Arbeitswelt vorbereiten? Und wie steht es um Werte wie Transparenz, Kommunikation auf Augenhöhe, Akzeptanz und Wertschätzung? Definieren sie nicht wichtige Rahmenbedingungen, wie gelehrt und gelernt werden sollte?
Multi-user virtual reality is transforming towards a social activity that is no longer only used by remote users, but also in large-scale location-based experiences. Usage of realtime-tracked avatars in co-located business-oriented applications with a ”guide-user-scenario” is examined for user-related factors of Spatial Presence, Social Presence, User Experience and Task Load. A user study was conducted in order to compare both techniques of a realtime-tracked avatar and a non-visualised guide. Results reveal that the avatar-guide enhanced and stimulated communicative processes while facilitating interaction possibilities and creating a higher sense of mental immersion for users and engagement.
We report about several experiments on applying mixed reality technology in the context of accessing collective memories from atomic bombs, Holocaust and Second World War. We discuss the impact of Virtual Reality, Augmented Virtuality and Augmented Reality for specific memorial locations. We show how to use a virtual studio for demonstrating an augmented reality application for a specific location in a remote session within a video conference. Augmented Virtuality is used to recreate the local environment, thus providing a context and helping the participants recollect emotions related to a certain place. This technique demonstrates the advantages of using virtual (VR) and augmented (AR) reality environments for rapid prototyping and pitching project ideas in a live remote setting.
The ability to understand and predict the pressure losses of orifices is important in order to improve the air flow within the secondary air system. This experimental study investigates the behavior of the discharge coefficient for circular orifices with inlet cross flow which is a common flow case in gas turbines. Examples of this are at the inlet of a film cooling hole or the feeding of air to a blade through an orifice in a rotor disk. Measurements were conducted for a total number of 38 orifices, covering a wide range of length-to-diameter ratios, including short and long orifices with varying inlet geometries. Up to five different chamfer-to-diameter and radius-to-diameter ratios were tested per orifice length. Furthermore, the static pressure ratio across the orifice was varied between 1.05 and 1.6 for all examined orifices. The results of this comprehensive investigation demonstrate the beneficial influence of rounded inlet geometries and the ability to decrease pressure losses, which is especially true for higher cross flow ratios where the reduction of the pressure loss in comparison to sharp-edged holes can be as high as 54%. With some exceptions, the chamfered orifices show a similar behavior as the rounded ones but with generally lower discharge coefficients. Nevertheless, a chamfered inlet yields lower pressure losses than a sharp-edged inlet. The obtained experimental data were used to develop two correlations for the discharge coefficient as a function of geometrical as well as flow properties.
We had already demonstrated a numerical model for the point spread function (PSF) of an optical system that can efficiently model both the experimental measurements and the lens design simulations of the PSF. The novelty lies in the portability and the parameterization of this model, which allow for completely new ways to validate optical systems, which is especially interesting not only for mass production optics such as in the automotive industry but also for ophthalmology. The numerical basis for this model is a nonlinear regression of the PSF with an artificial neural network (ANN). After briefly describing both the principle and the applications of the model, we then discuss two optically important aspects: the spatial resolution and the accuracy of the model. Using mean squared error (MSE) as a metric, we vary the topology of the neural network, both in the number of neurons and in the number of hidden layers. Measurement and simulation of a PSF can have a much higher spatial resolution than the typical pixel size used in current camera sensors. We discuss the influence this has on the topology of the ANN. The relative accuracy of the averaged pixel MSE is below 10 − 4, thus giving confidence that the regression does indeed model the measurement data with good accuracy. This article is only the starting point, and we propose several research avenues for future work.
Training autonomous vehicles requires lots of driving sequences in all situations. Collecting and labeling these drive scenes is a very time-consuming and expensive process. Currently, it is not possible to reuse these drive scenes with different optical properties, because there exists no numerically efficient model for the transfer function of the optical system. We present a numerical model for the point spread function (PSF) of an optical system that can efficiently model both experimental measurements and lens design simulations of the PSF. The numerical basis for this model is a nonlinear regression of the PSF with an artificial neural network. The novelty lies in the portability and the parameterization of this model. We present a lens measurement series, yielding a numerical function for the PSF that depends only on the parameters defocus, field, and azimuth. By convolving existing images and videos with this PSF, we generate images as if seen through the measured lens. The methodology applies to any optical scenario, but we focus on the context of autonomous driving, where the quality of the detection algorithms depends directly on the optical quality of the used camera system. With this model, it is possible to reuse existing recordings, with the potential to avoid millions of test drive miles. The parameterization of the optical model allows for a method to validate the functional and safety limits of camera-based advanced driver assistance systems based on the real, measured lens actually used in the product.
Der Lehre an deutschen Hochschulen stehen, neben einer angespannten Finanzlage, nun auch die Herausforderungen gegenüber, die sich unmittelbar aus dem doppelten Abitur-Jahrgang ergeben (KMK, 2012; BMBF, 2013). Um den hieraus resultierenden Anforderungen besser zu entsprechen, liegt es nahe, sich intensiv mit digital gestützten Lehr-/Lernkonzepten zu befassen. Hierzu zählen z.B. auch digitale Lehrmaterialien, die, unter besonderer Berücksichtigung mobiler Endgeräte (Bitkom, 2012), in Form von interaktiven Lernmodulen distribuiert werden. Im Rahmen der eStudy-Aktivitäten des Fachbereichs Medien der FH Düsseldorf wurde eine Autorenumgebung konzipiert, die eine hocheffiziente Entwicklung von HTML5-basierten Lernmodulen mit interessanten Zusatzfunktionen erlaubt, die in vielfältigen Lehr-/Lernszenarien zum Einsatz kommen können.
Einfach benutzbare VR-Anwendungen erfordern andere Interaktionstechniken als konventionelle Desktop-Anwendungen mit Maus, Tastatur und Desktop-Metapher zur Verfügung stellen. Da solche Ansätze in Konzeption und Realisierung deutlicher komplexer sind, müssen diese mit Sorgfalt ausgewählt werden. Folgt man der Argumentation, dass VR eine natürliche Interaktion mit virtuellen Objekten ermöglicht, so führt dies fast zwangsläufig zu zweihändigen Interaktionstechniken für virtuelle Umgebungen, da Benutzer in realen Umgebungen gewohnt sind, fast ausschlieï‚lich zweihändig zu agieren. In diesem Beitrag geben wir eine Übersicht über den Stand der Technik im Bereich zweihändiger Interaktion, leiten Anforderungen an eine Entwicklung zweihändiger Interaktionstechniken in VR ab und beschreiben einen eigenen Ansatz. Dabei geht es um die zweihändige Interaktion bei der Simulation flexibler biegeschlaffer Bauteile (z. B. Schlauchverbindungen).