Microfluidic Platforms for the Investigation of Intercellular Signalling Mechanisms

Intercellular signalling has been identified as a highly complex process, responsible for orchestrating many physiological functions. While conventional methods of investigation have been useful, their limitations are impeding further development. Microfluidics offers an opportunity to overcome some of these limitations. Most notably, microfluidic systems can emulate the in‐vivo environments. Further, they enable exceptionally precise control of the microenvironment, allowing complex mechanisms to be selectively isolated and studied in detail. There has thus been a growing adoption of microfluidic platforms for investigation of cell signalling mechanisms. This review provides an overview of the different signalling mechanisms and discusses the methods used to study them, with a focus on the microfluidic devices developed for this purpose.     

Antagonistic Action of Fe and Mn in Mediterranean-Type Plants Irrigated with Wastewater Effluents Following Biological Treatment

The present study examined the ability of certain Mediterranean plants to absorb Mn ++ when irrigated with wastewater effluents following treatment, the relative absorption of Fe ++ and Mn ++ , as well as the endurance of such plants to high Mn ++ concentrations in the water used for their irrigation. The experimental plants used in the study were Olea europaea , Nerium oleander and Geranium sp . These plants were irrigated with secondary treatment wastewater effluents, with water commonly used for irrigation in the area, and with water enriched with high concentrations of Mn ++ . The results showed that irrigation of the plants with wastewater treatment effluents resulted in slightly elevated concentrations of Mn ++ and Fe ++ in the leaves, the roots, and in the soil. On the contrary, irrigation with water enriched with high concentrations of Mn ++ (as high as 1000 mg/L) resulted in correspondingly high Mn ++ concentrations in the plants under study. The accumulation of Mn ++ was much higher in the roots than in the leaves of these plants. In the case of Mn ++ enriched water irrigation, the corresponding concentrations of Fe ++ in the roots, in the leaves, and in the soil indicated the existence of strong antagonism between these two elements. All plants exhibited tolerance to high concentrations of Mn ++ .     

Mitochondrial alterations and autofluorescent conversion of Candida albicans induced by histatins

The mechanism of the candidacidal activity of histatins 3 and 5 (Hst) is still a matter of debate. Previous studies have indicated that Hst induce cell permeabilization, generation of reactive oxygen species (ROS) by mitochondria, inhibition of the respiratory chain, and energy-dependent cytotoxic release of ATP. On the other hand, the multiplicity of effects and the apparent contrast between experimental data continue to render the mechanism of Hst-induced killing of C. albicans unclear. In this investigation, using fluorescent probes (the potential-sensitive mitochondrial probe tetramethylrhodamine methyl ester perchlorate, TMRM; the ROS-sensitive probe dihydrofluorescein diacetate, DHF; the membrane-impermeant probe, calcein) and autofluorescence data we observed that Hst induce ROS generation by mitochondria undergoing a high energy swelling condition, accompanied by oxidation of cytosolic NAD(P)H and mitochondrial flavoproteins. ROS generation and swelling, attributable to an inhibition of the respiratory chain and to impairment of the K/H-exchanger, were followed by mitochondrial depolarization. Mitochondrial changes were accompanied by massive calcein influx, indicative of cell permeabilization, and prominent alterations of the cell size, shape, and optical density. The loss of proliferative activity was correlated, on a single cell basis, to the acquisition of a lipofuscin-like autofluorescence.     

Actuation of Three-Dimensional-Printed Nanocolloidal Hydrogel with Structural Anisotropy

Polymer hydrogels exhibit actuation properties that result in reversible shape transformations and have promising applications in soft robotics, drug delivery systems, sensors, and microfluidic devices. Actuation occurs due to differential hydrogel swelling and is generally achieved by modulating hydrogel composition. Here a different approach to hydrogel actuation that originates solely from its structural anisotropy is reported. For 3D-printed single-layer hydrogels formed by cellulose nanocrystals (CNCs) and gelatin methacryloyl it is shown that shear-induced orientation of CNCs results in anisotropic mechanical and swelling properties of the hydrogel. Upon swelling in water, planar hydrogels acquire multiple complex 3D shapes that are achieved by i) varying CNC orientation with respect to the shape on the hydrogel sheet and ii) patterning the hydrogel with the regions of shear-mediated and random CNC orientation. This study shows the capability to generate multiple shapes from the same hydrogel actuator based on the degree of its structural anisotropy. In addition, it introduces a biocompatible nanocolloidal ink with shear-thinning and self-healing properties for additive manufacturing of hydrogel actuators.     

Cybersecurity investments with nonlinear budget constraints and conservation laws: variational equilibrium,marginal expected utilities,and Lagrange multipliers

In this paper, we propose a new cybersecurity investment supply chain game theory model, assuming that the demands for the product are known and fixed and, hence, the conservation law of each demand market is fulfilled. The model is a generalized Nash equilibrium model with nonlinear budget constraints for which we define the variational equilibrium, which provides us with a variational inequality formulation. We construct an equivalent formulation, enabling the analysis of the influence of the conservation laws and the importance of the associated Lagrange multipliers. We find that the marginal expected transaction utility of each retailer depends on this Lagrange multiplier and its sign. Finally, numerical examples with reported equilibrium product flows, cybersecurity investment levels, and Lagrange multipliers, along with individual firm vulnerability and network vulnerability, illustrate the obtained results.     

The prebiotic effect of α-1,2 branched,low molecular weight dextran in the batch and continuous faecal fermentation system

The aim of this study was to establish the effect of smaller molecular weight (0.5 and 1.0 kDa) on prebiotic efficacy and its putative sustainability in the human gut. The prebiotic effect of α-1,2 branched, 0.5 and 1 kDa dextrans were evaluated in faecal batch fermentations as compared with inulin. Both dextrans induce similar selectivity towards Bifidobacterium sp., Lactobacillus/Enterococcus and Bacteroides/Prevotella, and producing similar concentrations of short chain fatty acids. However, the 0.5 kDa dextran was fermented faster than the 1 kDa dextran, where both produced lower amount of gas than inulin. The fermentation of 1 kDa dextran was further investigated in continuous gut models. The dextran increased Bifidobacterium and Roseburia sp. populations in the final vessel, while decreasing Clostridium histolyticum and Faecalibacterium prausnitzii. Overall, the α-1,2 branched, 1 kDa dextran induced selective effect on the gut microbiota and stimulated short chain fatty acids, indicating prebiotic sustainability in distal regions of the gut.     

Determination of ochratoxin A in grapes of Greek origin by immunoaffinity and high-performance liquid chromatography

A simple analytical method for ochratoxin A (OTA) determination in grapes is described, using aqueous methanolic extraction, an immunoaffinity column clean-up step and high-performance liquid chromatography with fluorescence detection. Mean recovery was 94% (RSD = 4.0%) with a detection limit of 0.4 ng g^-1 and quantification limit of 1.20 ng g^-1. Repeatability (r) and reproducibility (R) were 1.17 and 1.34, respectively. OTA determinations were applied to 50 grape samples (23 different varieties) originating from representative regions of Greece. Results showed the presence of OTA in 86% of samples tested (n = 50). Traces were found in 56% of samples but OTA was not detectable in 14% of samples. Traces were also found in 4% of red, organically grown samples. The most contaminated were three samples of red grapes, two from Central Greece (2.69 and 1.41 ng g^-1), both table and wine-making grapes. The third sample (1.46 ng g^-1), originating from the island of Samos, was used only in wine-making. Mean (1.06 ng g^-1) and median (0.76 ng g^-1) OTA concentrations in red grapes were slightly higher compared to the mean (0.82 ng g^-1) and median (0.65 ng g^-1) concentrations in white grape samples. The study shows that the potential risk for a person of 60 kg ranged from 0.9 to 9 ng kg^-1 bw day^-1 and is dependent on the quantity of grapes consumed daily.     

Fatty acid and carotenoid composition of egg yolk as an effect of microalgae addition to feed formula for laying hens

Hens were fed five diets based on rapeseed/corn oils with or without addition of the marine microalgae, Nannochloropsis oculata. Colours of egg yolk, fatty acid profiles in phospholipid (PL) and triacylglycerol (TAG) fractions and carotenoid contents were analysed. The major effects on yolk fatty acid composition were observed in the PL fraction with changes in 18:2n − 6, 18:3n − 3, 20:4n − 6, 20:5n − 3 and 22:6n − 3. The highest amounts of eicosapentaenoic acid and docosahexaenoic acid were obtained from the diet containing 20% N. oculata, whereas the highest arachidonic acid content in yolk was from the corn oil diet. In addition, colour (a* value) and carotenoid content increased when N. oculata was included in the diet. Long chained fatty acids were almost exclusively present in the PL fraction of yolk. This study shows that fatty acid composition and carotenoid content of egg yolk are improved by addition of N. oculata in laying hen diet.     

The effect of moisture on the dynamical mechanical properties of bacterial cellulose/glucuronoxylan nanocomposites

The plant cell wall possesses unique material properties due to its hierarchical organisation. In order to biomimic a native structure like a plant cell wall, a model system consisting of microfibrillar cellulose, produced by the gram-negative bacteria Acetobacter xylinum, and a glucuronoxylan matrix derived from aspen holocellulose was constructed. The glucuronoxylan was extracted from delignified aspen (Populus tremula) wood chips using DMSO to preserve its native chemical composition. Dynamic mechanical analysis (DMA) measurements performed with moisture scans showed a moisture-induced softening of delignified aspen wood fibres due to the plasticization of glucuronoxylan. A similar result was observed for the model system. However, the softening behaviour of the delignified aspen fibre and the model system was not identical, most probably due to differences in spatial organisation of the components. Dynamic FTIR-studies indicated that interactions between the cellulose and the glucuronoxylan exist in the aspen holocellulose while the components in the nanocomposite appear to be more isolated.     

Safety evaluation of slender high-strength concrete columns under sustained loads

A realistic analysis of the reliability of slender high-strength concrete (HSC) columns must include the long-term effects of sustained loads. In this paper, the long-term reliability of eccentrically loaded slender HSC columns under sustained loads is evaluated. The reliability of slender reinforced concrete columns representing normal-strength concrete and HSC is investigated. Since most of the variables involved in column design (material properties, geometric characteristics, loads, etc.) are random, probabilistic methods are used in the analysis. The effects of the concrete compressive strength, amount of longitudinal steel, load eccentricity, and slenderness ratio on the column reliability are investigated. It was found that all these factors have a considerable impact on the resulting column reliability.