Biosensor Based on Peroxidase-Mimetic Nanozyme and Lactate Oxidase for Accurate L-Lactate Analysis in Beverages.

Precision analysis of the key biological metabolites such as L-lactate has great practical importance for many technological processes in food technology, including beverage production. Here we describe a new, highly selective, and sensitive biosensor for accurate L-lactate assay based on a combination of peroxidase-mimetic nanozymes with microbial lactate oxidase (LOx) immobilized onto the surface of a graphite-rod electrode (GE). The peroxidase-like nanozymes were synthesized using the debris of carbon microfibers (CFs) functionalized with hemin (H) and modified with gold nanoparticles (AuNPs) or platinum microparticles (PtMPs). The nanozyme formed with PtMPs as well as corresponding bioelectrodes based on it (LOx-CF-H-PtMPs/GE) is characterized by preferable catalytic and operational characteristics, so it was selected for the analysis of L-lactate content in real samples of grape must and red wine. The results of the L-lactate analysis obtained by the developed biosensors are highly correlated with a very selective spectrophotometric approach used as a reference. The developed biosensor, due to its high selectivity and sensitivity, is very prospective not only for the beverage industry and food technology, but also for clinical diagnostics and medicine, as well as in other applications where the accurate analysis of L-lactate is highly important.

On crystallization of water confined in liposomes and cryoprotective action of DMSO.

In this work, the phase behavior of cryoprotective mixtures based on dimethyl sulfoxide (DMSO) mixed with a lipid bilayer consisting of dimyristoyl phosphatidylcholine (DMPC) was studied. This system represented a model of a biological cell and its membrane. The aim of the work was to clarify the origin of the cryoprotective action of low-concentrated mixtures (1-10 vol%) DMSO in water, representing mixtures used in cryopreservation in cell therapy. The combination of experimental techniques of differential scanning calorimetry (DSC) and positron annihilation lifetime spectroscopy (PALS) allowed a study of crystallization behavior of water confined in liposomes imitating the intracellular environment. The ability of liposomes to show the fundamental aspects of water phase behavior seen during freezing of biological cells was proved. The presence of an amorphous freeze-concentrated phase of DMSO in the frozen state was confirmed and its possible crystallization into the DMSO trihydrate and ice during thawing was demonstrated. Correlation between the critical temperature range for the loss of cell viability during slow thawing and the temperatures of freeze-concentrated phase crystallization was found. Based on this finding, possible mechanisms of DMSO cryoprotection are discussed with support brought by results for the studied model system. Quantification of the ice phase fraction in the frozen mixtures revealed that even low concentrations of DMSO can induce a considerable decrease in the amount of ice present.

Microporous carbon fibers as electroconductive immobilization matrixes: Effect of their structure on operational parameters of laccase-based amperometric biosensor.

In this study, we describe the fabrication of sensitive biosensor for the detection of phenolic substrates using laccase immobilized onto two types of microporous carbon fibers (CFs). The main characteristics of microporous CFs used for preparation of biosensors are given. Two CFs were characterized by different specific surface area, CFA (<1 m2·g-1) and CFB (1448 m2·g-1), but with comparable size of the micropores estimated by positron annihilation lifetime spectroscopy. The structural analysis was shown that CFA is formed by thin interwoven fibers forming a highly porous structure, as well as CFB - by granular formations with uneven edges that shape a cellulose membrane of lower porosity. The results of amperometric analysis revealed that the laccase-bound CFs possesses better electrochemical behavior for laccase than non-modified rod carbon electrodes (control). Using chronoamperometric analysis, the operational parameters of the CFs-modified bioelectrodes were compared to control bioelectrodes. The bioelectrodes based on CFs have demonstrated 2.4-2.7 folds enhanced maximal current at substrate saturation (Imax) values, 1.2-1.4 folds increased sensitivity and twice wide linearity compared with control bioelectrodes. The sensitivity of the developed CFs-based bioelectrodes was improved compared with the laccase-bound electrodes, described in literature. The developed biosensor was tested for catechol analysis in the real communal wastewater sample.

Microstructural free volume and dynamics of cryoprotective DMSO-water mixtures at low DMSO concentration.

This work investigates the free-volume properties of the dimethyl sulfoxide (DMSO)-water mixtures by positron annihilation lifetime spectroscopy over a wide temperature range of 20-320 K. The processes of melting and solidification of the water, DMSO and the DMSO-water mixtures at 1.8, 2.0 and 10% vol. DMSO respectively were studied. It was found that the recrystallization during heating of the water-DMSO cryoprotective mixtures above 160 K at low DMSO concentrations is affected by the amount of DMSO in the mixture. The amount of amorphous phase formed during cooling influences the hysteresis between cooling and heating cycles which could be crucial for cell survival. Experiments also show the time dependence of crystallization which indicates that rapid heating can suppress this secondary crystallization which is undesirable during the cell thawing process. Similar concentrations of DMSO (1.8% and 2% vol. DMSO in water) where a 2% vol. DMSO mixture secures cell survival but 1.8 vol% does not, showed differences in structural and dynamic properties that are key factors in cell survival. These results were supported by differential scanning calorimetry and low frequency dielectric spectroscopy measurements. The obtained data are in strong agreement with the observed cryoprotective efficacy of the DMSO-water mixtures on living cells.

Preparation and characterization of MnO2- SiO2 composite resin for 226Ra pre-concentration in water samples.

The presented work describes an effective method for 226Ra determination using laboratory- prepared MnO2-SiO2 composite resin. Samples were traced with 133Ba for radium radiochemical recoveries monitoring which were in the range of (77 - 100%). MnO2- SiO2 composite resin was used to collect for 226Ra from water samples (pH = 6.5-7.0). Radium was eluted from the column with 50 mL of 4 mol L-1 HNO3 at a flow rate of 1.5 mL min-1. Alpha-spectrometric counting of Ba(Ra)SO4 microprecipitate was used for 226Ra determination. The new developed method was used to the 226Ra determination in samples of natural mineral, mountain spring, drinking and natural healing waters from Slovakia and Czech Republic.

Cross-sectional TEM study of subsurface damage in SPDT machining of germanium optics.

In addition to surface roughness and shape precision, the subsurface damage (SSD) generated by single point diamond turning (SPDT) of Ge and Si crystal optics is of increasing importance with decreasing wavelength from infrared through visible, UV, and x-ray. There are various components of SSD, e.g., microcracks, dislocations, strain, and a near-surface amorphous layer, and there are also several techniques to evaluate various components of SSD. Cross-sectional transmission electron microscopy (XTEM) is expensive and not often directly used in the optics laboratory. However, because of its very high sensitivity to SSD and down to atomic resolution, it is often used as an external service for developing SPDT technology and other surface processing techniques. It is shown in the paper that improper sample preparation can generate near-surface amorphization. Measures to avoid this artifact and a test of reliability of XTEM sample preparation are proposed.