Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

Strawberry juice colour: The effect of some processing variables on the stability of anthocyanins

The colour stability of juice and purées made from a mixture of two strawberry varieties, stored at + 20°C or at –20°C under aerobic and anaerobic conditions, was studied. High-performance liquid chromatography and spectrophotometric analysis were used to monitor the changes in colour during storage. Pelargonidin 3-glucoside comprised 80% of the total anthocyanin content. Four other pelargonidin-based peaks were found, being 10.4%, 3.5%, 0.7% and 0.3% of the total and two cyanidin peaks were present, being 3.3% and 0.4% of the total. Clarification before storage caused considerable losses in the initial anthocyanin concentrations. Storage conditions (air versus nitrogen) did not influence the rate of loss of anthocyanins during storage or the formation of polymeric pigments. There were no losses in anthocyanins and no increase in the amount of colour measured at 510 nm due to polymers in samples stored at–20°C. Polymerisation occurred in the samples stored at + 20°C, especially in the clarified samples.     

X-ray Fluorescence Uptake Measurement of Functionalized Gold Nanoparticles in Tumor Cell Microsamples

Quantitative cellular in vitro nanoparticle uptake measurements are possible with a large number of different techniques, however, all have their respective restrictions. Here, we demonstrate the application of synchrotron-based X-ray fluorescence imaging (XFI) on prostate tumor cells, which have internalized differently functionalized gold nanoparticles. Total nanoparticle uptake on the order of a few hundred picograms could be conveniently observed with microsamples consisting of only a few hundreds of cells. A comparison with mass spectroscopy quantification is provided, experimental results are both supported and sensitivity limits of this XFI approach extrapolated by Monte-Carlo simulations, yielding a minimum detectable nanoparticle mass of just 5 pg. This study demonstrates the high sensitivity level of XFI, allowing non-destructive uptake measurements with very small microsamples within just seconds of irradiation time.     

Suppression of the development of pores during laser-induced surface dispersion of TiC into aluminium, by means of a static magnetic field

A static magnetic field having the strength of B=0.6 T has been used as an additional process parameter in order to suppress the establishment of pores while dispersing TiC particles in surface layers of aluminium alloys using a laser. The migration of hydrogen towards regions of high temperatures within the melting pool can lead to the creation of gas bubbles, therefore yielding pores after resolidification in the processed layer. This mechanism (known for instance in welding technologies of aluminium) could be suppressed by Lorentz-forces induced by an externally applied magnetic field. It could be shown experimentally that magnetic fields could assist laser surface treatment processes beneficially if higher beam powers were coupled in. This may be interesting if, for instance, thicker dispersed layers should be achieved.     

The influence of training level on manual flight in connection to performance,scan pattern,and task load

This work focuses on the analysis of pilots’ performance during manual flight operations in different stages of training and their influence on gaze strategy. The secure and safe operation of air traffic is highly dependent on the individual performances of the pilots. Before becoming a pilot, he/she has to acquire a broad set of skills by training to pass all the necessary qualification and licensing standards. A basic skill for every pilot is manual control operations, which is a closed-loop control process with several cross-coupled variables. Even with increased automation in the cockpit, the manual control operations are essential for every pilot as a last resort in the event of automation failure. A key element in the analysis of manual flight operations is the development over time in relation to performance and visual perception. An experiment with 28 participants (including 11 certified pilots) was conducted in a Boeing 737 simulator. For defined flight phases, the dynamic time warping method was applied to evaluate the performance for selected criteria, and eye-tracking methodology was utilized to analyze the gaze-pattern development. The manipulation of workload and individual experience influences the performance and the gaze pattern at the same time. Findings suggest that the increase of workload has an increased influence on pilots depending on the flight phase. Gaze patterns from experienced pilots provide insights into the training requirements of both novices and experts. The connection between workload, performance and gaze pattern is complex and needs to be analyzed under as many differing conditions. The results imply the necessity to evaluate manual flight operations with respect to more flight phases and a detailed selection of performance indications.

     

Future Proof: The Transition to Digital at IFI Archive

Informed by research over the past two years and the work of colleagues in peer institutions, the IFI Irish Film Archive developed a six-year Digital Preservation and Access strategy which launched in 2014. Fundamental to this Strategy was the design and installation of digital archive tools for long term preservation and the redesign of workflow practices to facilitate accession and management of high resolution digital film and broadcast assets and associated metadata. This article outlines the steps taken and standards applied in developing a future proof digital audiovisual archive.     

Cellular analysis by open-source software for affordable cytometry

Image cytometry is an important technique in affordable healthcare and cellular research. Some efforts toward establishing a personal, low-cost cytometer have been described in the literature. However, a self-assembled fluorescence microscope requires software for cytometric analysis. There are some open-source image-based software analysis applications available. However, for a quantitative analysis of images, software that can generate data comparable to those of previously evaluated cytometric analyses programs is required. Hence, the aim of this study is to compare results of a commercially available image cytometry program to data obtained using the open-source software CellProfiler (CP). Leukocytes and fluorescent bead images obtained using a Laser Scanning Cytometer were analyzed by CP and the results compared with those of conventional cytometric analyses' programs. Algorithms were developed enabling the analysis of leukocytes and beads by CP. CP provided similar results to those obtained by the cytometer software. Hallmark parameters, including cell count and fluorescence intensity, revealed a high correlation in the analysis of both programs. Therefore, CP is appropriate for cellular analysis on a self-assembled microscope, thereby enabling affordable cytometry.     

Workflows for Heliophysics

In this paper we describe how we have introduced workflows into the working practices of a community for whom the concept of workflows is very new, namely the heliophysics community. Heliophysics is a branch of astrophysics which studies the Sun and the interactions between the Sun and the planets, by tracking solar events as they travel throughout the Solar system. Heliophysics produces two major challenges for workflow technology. Firstly it is a systems science where research is currently developed by many different communities who need reliable data models and metadata to be able to work together. Thus it has major challenges in the semantics of workflows. Secondly, the problem of time is critical in heliophysics; the workflows must take account of the propagation of events outwards from the sun. They have to address the four dimensional nature of space and time in terms of the indexing of data. We discuss how we have built an environment for Heliophysics workflows building on and extending the Taverna workflow system and utilising the myExperiment site for sharing workflows. We also describe how we have integrated the workflows into the existing practices of the communities involved in Heliophysics by developing a web portal which can hide the technical details from the users, who can concentrate on the data from their scientific point of view rather than on the methods used to integrate and process the data. This work has been developed in the EU Framework 7 project HELIO, and is being disseminated to the worldwide Heliophysics community, since Heliophysics requires integration of effort on a global scale.     

Making it easier for older people to talk to smart homes: the effect of early help prompts

It is well known that help prompts shape how users talk to spoken dialogue systems. This study investigated the effect of help prompt placement on older users’ interaction with a smart home interface. In the dynamic help condition, help was only given in response to system errors; in the inherent help condition, it was also given at the start of each task. Fifteen older and sixteen younger users interacted with a smart home system using two different scenarios. Each scenario consisted of several tasks. The linguistic style users employed to communicate with the system (interaction style) was measured using the ratio of commands to the overall utterance length (keyword ratio) and the percentage of content words in the user’s utterance that could be understood by the system (shared vocabulary). While the timing of help prompts did not affect the interaction style of younger users, it was early task-specific help supported older users in adapting their interaction style to the system’s capabilities. Well-placed help prompts can significantly increase the usability of spoken dialogue systems for older people.