Comparative Analysis of Platelet-Derived Extracellular Vesicles Using Flow Cytometry and Nanoparticle Tracking Analysis

Growing interest in extracellular vesicles (EVs) has prompted the advancements of protocols for improved EV characterization. As a high-throughput, multi-parameter, and single particle technique, flow cytometry is widely used for EV characterization. The comparison of data on EV concentration, however, is hindered by the lack of standardization between different protocols and instruments. Here, we quantified EV counts of platelet-derived EVs, using two flow cytometers (Gallios and CytoFLEX LX) and nanoparticle tracking analysis (NTA). Phosphatidylserine-exposing EVs were identified by labelling with lactadherin (LA). Calibration with silica-based fluorescent beads showed detection limits of 300 nm and 150 nm for Gallios and CytoFLEX LX, respectively. Accordingly, CytoFLEX LX yielded 40-fold higher EV counts and 13-fold higher counts of LA⁺CD41⁺ EVs compared to Gallios. NTA in fluorescence mode (F-NTA) demonstrated that only 9.5% of all vesicles detected in scatter mode exposed phosphatidylserine, resulting in good agreement of LA⁺ EVs for CytoFLEX LX and F-NTA. Since certain functional characteristics, such as the exposure of pro-coagulant phosphatidylserine, are not equally displayed across the entire EV size range, our study highlights the necessity of indicating the size range of EVs detected with a given approach along with the EV concentration to support the comparability between different studies.     

Effects of upholstery materials on the burning behavior of real‐scale residential upholstered furniture mock‐ups

Fire spread and growth on real‐scale four cushion mock‐ups of residential upholstered furniture (RUF) were investigated with the goal of identifying whether changes in five classes of materials (barrier, flexible polyurethane foam, polyester fiber wrap, upholstery fabric, and sewing thread), referred to as factors, resulted in statistically significant changes in burning behavior. A fractional factorial experimental design plus practical considerations yielded a test matrix with 20 material combinations. Experiments were repeated a minimum of two times. Measurements included fire spread rates derived from video recordings and heat release rates (HRRs). A total of 13 experimental parameters (3 based on the videos and 10 on the HRR results), referred to as responses, characterized the measurements. Statistical analyses based on Main Effects Plots (main effects) and Block Plots (main effects and factor interactions) were used. The results showed that three of the factors resulted in statistically significant effects on varying numbers of the 13 responses. The Barrier and Fabric factors had the strongest main effects with roughly comparable magnitudes. Foam was statistically significant for fewer of the responses and its overall strength was weaker than for Barrier and Fabric. No statistically significant main effects were identified for Wrap or Thread. Multiple two‐term interactions between factors were identified as being statistically significant. The Barrier*Fabric interaction resulted in the highest number of and strongest statistically significant effects. The existence of two‐term interactions means that it will be necessary to consider their effects in approaches designed to predict the burning behavior of RUF.     

Development and characterisation of a Y2Ti2O7-based glass-ceramic as a potential oxidation protective coating for titanium suboxide (TiOx)

A silica-based glass-ceramic, with Y₂Ti₂O₇ as the major crystalline phase, is designed, characterised and tested as an oxidation-protective coating for a titanium suboxide (TiO_x) thermoelectric material at temperatures of up to 600 °C. The optimised sinter-crystallisation treatment temperatures are found to be 1300 °C and 855 °C for a duration of 30 min, and this treatment leads to a glass-ceramic with cubic Y₂Ti₂O₇ and CaAl₂Si₂O₈ as crystalline phases. An increase of ~270 °C in the dilatometric softening temperature is observed after devitrification of the parent glass, thus further extending its working temperature range.Excellent adhesion of the glass-ceramic coating to the thermoelectric material is maintained after exposure to a temperature of 600 °C for 120 h under oxidising conditions, thus confirming the effectiveness of the T1 glass-ceramic in protecting the TiO_x material.     

Influence of the Impaction Angle on the Triboelectric Charging of Aerosol Nanoparticles

A low pressure impactor is used to measure triboelectric charging behavior of metallic nanoparticles. Ag nanoparticles, produced by spark discharge, were impacted onto Pt sputtered targets. The influence of the impaction angle and impaction velocity on the triboelectric charging was investigated. While for perpendicular impaction the charge transfer behavior of previous work was confirmed, the oblique impaction revealed new phenomena. Additional charge transfer was observable, which increases with obliqueness. The possibility of mass transfer between particle and target due to the high-energy collisions was also investigated. SEM characterization and Auger spectroscopy indicate mass transfer from the particle to the target surface.     

Novel pearl millet couscous process for West African markets using a low-cost single-screw extruder

Traditional West African pearl millet couscous products are popular; however, their preparation is laborious, time-consuming and energy-demanding, involving agglomeration, steaming, drying and sieving steps. In this study, a process was developed to produce millet couscous using a high pressure, high temperature and low-cost single-screw extruder. The innovation was to directly process the cooked low-moisture extrudate to a couscous product by drying and milling to the appropriate particle size (ranging between 1 and 2 mm). Throughput for the traditional process as prepared for commercial sale is 50 kg per day, but with the same amount of labour, the extrusion process yields ~350 kg per day. A consumer sensory study held in Niamey (Niger) showed that the extruded millet couscous was comparable to traditional couscous, though this was dependent on selection of the proper millet variety. This novel extrusion process could stimulate small- to medium-scale manufacturing of couscous and couscous-type products in West Africa.     

Furan fatty acid concentrations in tea infusions prepared from green,black, and herbal teas

Furan fatty acids (FuFAs) are valuable antioxidants with highly effective radical scavenging properties which are widely distributed at low levels in food. Previous research indicated that tea is a valuable source of FuFAs. However, tea is only consumed in form of infusions. To fill this gap, we prepared infusions from different herbal, green, and black teas. Initial measurements with GC-MSMS of tea verified previous findings that 11-(3,4-dimethyl-5-penylfuran-2-yl)-undecanoic acid (11D5) was the prevalent FuFA in tea matrix. Therefore, 11D5 was quantified in tea infusions by means of UHPLC-MSMS equipment after mild alkaline hydrolysis. While herbal tea infusions were low or free of FuFAs, 11D5 was detectable in all samples of green and black tea infusions. Amounts of 11D5 were higher in green tea than in black tea. Moreover, Darjeeling tea infusions were by ~30% richer in 11D5 than black and green teas from other regions. Each cup of green and black tea infusion may provide 20–60 μg 11D5, which is about 5% of the amounts found in tea samples. Spread over the day, regular tea consumption may contribute to the intake of valuable FuFAs.     

5G MEC-enabled vehicle discovery service for streaming-based CAM applications

Cooperative perception represents an important technology to fulfil the higher automation levels of connected and automated mobility (CAM). In cooperative perception, the sensor data, either raw or processed, is shared among neighbour vehicles with the objective of enhancing or complementing the perception obtained by on-board sensors. The vehicle that requests this external perception data needs to have this data quickly. However, it first needs to discover the network address of the neighbour vehicle that wants to connect to. Specially in a dense urban area or in a congested radio channel, an inefficient method for neighbour vehicle discovery could prevent a timely start of the cooperative perception session. This paper describes a novel 5G multi-access edge computing (MEC) solution that that boosts the selection of interesting neighbour vehicles according to a geographical region of interest (ROI) after applying pertinent adjustments considering vehicles’ dynamics and network communication latencies. In contrast to broadcast-based methods, in the proposed method the vehicles are only sending their periodical position data to a MEC service, which centralises the vehicle discovery requests. The objective of this Vehicle Discovery Service (VDS) is to support the startup of Web Real-Time Communications (WebRTC)-based Extended Sensors CAM applications. The proposed VDS has been validated using a public vehicular traffic dataset evaluating geo-position accuracy. The WebRTC-based streaming pipeline has been validated testing its feasibility for a See-Through video streaming application.

     

N-Heterocyclization in Gliotoxin Biosynthesis is Catalyzed by a Distinct Cytochrome P450 Monooxygenase

Gliotoxin and related epidithiodiketopiperazines (ETP) from diverse fungi feature highly functionalized hydroindole scaffolds with an array of medicinally and ecologically relevant activities. Mutation analysis, heterologous reconstitution, and biotransformation experiments revealed that a cytochrome P450 monooxygenase (GliF) from the human-pathogenic fungus Aspergillus fumigatus plays a key role in the formation of the complex heterocycle. In vitro assays using a biosynthetic precursor from a blocked mutant showed that GliF is specific to ETPs and catalyzes an unprecedented heterocyclization reaction that cannot be emulated with current synthetic methods. In silico analyses indicate that this rare biotransformation takes place in related ETP biosynthetic pathways.     

Production of the Carboxylate Reductase from Nocardia otitidiscaviarum in a Soluble,Active Form for in vitro Applications

Accessing aldehydes from carboxylate moieties is often a challenging task. In this regard, carboxylate reductases (CARs) are promising catalysts provided by nature that are able to accomplish this task in just one step, avoiding over-reduction to the alcohol product. However, the heterologous expression of CARs can be quite difficult due to the excessive formation of insoluble protein, thus hindering further characterization and application of the enzyme. Here, the heterologous production of the carboxylate reductase from Nocardia otitidiscaviarum (NoCAR) was optimized by a combination of i) optimized cultivation conditions, ii) post-translational modification with a phosphopantetheinyl transferase and iii) selection of an appropriate expression strain. Especially, the selection of Escherichia coli tuner cells as host had a strong effect on the final 110-fold increase in the specific activity of NoCAR. This highly active NoCAR was used to reduce sodium benzoate to benzaldehyde, and it was successfully assembled with an in vitro regeneration of ATP and NADPH, being capable of reducing about 30 mM sodium benzoate with high selectivity in only 2 h of reaction.