DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R² > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine.     

Production and Emulsifying Effect of Polyglycerol and Fatty Acid Esters with Varying Degrees of Esterification

Esters with acyl groups can be formed by the esterification of polyglycerol. The purpose of the present study was to produce fatty acid esters [hexanoic (caproic), octanoic (caprylic), decanoic (capric), dodecanoic (lauric), tetradecanoic (myristic), hexadecanoic (palmitic), octadecanoic (stearic)] and polyglycerol (average number‐of degrees of polymerization of 5) with varying degrees of esterification and to examine their emulsifying properties. A number of fundamental catalysts of polyglycerol acylation reactions by methyl esters of carboxylic acid were studied, and sodium methoxide was found to be the best choice. The temperature rate of transesterification increased from 180 to 220 °C with the fatty acid chain alkyl residue. Synthesized mono‐, di‐, tri‐, tetra‐, and heptaesters of various fatty acids and polyglycerol provided the highest hydroxyl values from 15 to 815 mg KOH g^?1 and saponification values from 82 to 321 mg KOH g^?1. The emulsifying properties were assessed for all polyglycerol and fatty acid esters, with results showing maximum emulsifying effect for tri‐ and tetraesters of capric, lauric, and caprylic acids. Regardless of the hydrophilic–lipophilic balance value (HLB) of polyglycerol esters and carboxylic acid, a 4:1 ratio of sunflower oil to water formed a water‐in‐oil type emulsion. When mixing oil and water in a 1:1 ratio, mono‐ and diesters of polyglycerol formed an oil‐in‐water type emulsion, heptaesters formed a water‐in‐oil type emulsion, and tri‐ and tetraesters formed both of types of emulsions, depending on the length of the acid hydrocarbon radicals.     

Interpolyelectrolyte complexes of maleic acid copolymers and chitosan for stabilization and functionalization of magnetite nano‐ and microparticles

A facile method of preparation of stabilized and functionalized nano‐ and microparticles of magnetite by successive application of oppositely charged polymers with a regular structure of macromolecular chains (chitosan and maleic acid copolymers) onto Fe₃O₄ core are developed. This approach makes it possible to create two types of magnetite interpolyelectrolyte shells, containing carboxylic or amino groups in outer layer of shell. Composition and magnetic properties of composite particles depend on nature of the copolymer of maleic acid, reaction conditions and size of obtained particles. The carboxylic groups of copolymer in outer layer of interpolyelectrolyte shell were converted into reactive anhydride groups by heating. Thermal treatment also leads to covalent crosslinking of shell and improves stability of composites both in acidic and alkaline media. Horseradish peroxidase was successfully immobilized onto covalently crosslinked and activated microparticles of Fe₃O₄ in aqueous medium without of condensing agents. The proposed reproducible and low‐cost technique does not use toxic reagents or solvents at all stages, including preparation of Fe₃O₄, formation, activation and crosslinking, of magnetite shell, modification of activated surface of composite particles. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39663.     

Influence of Macromolecular Brushes with Polyimide Backbones and Poly(methyl methacrylate) Side Chains on Structure,Physical, and Transport Properties of Polyphthalamide

Composite films based on polyamide (poly(m-phenylene-iso-phthalamide)) (PA) and a brush-like polymer with polyimide (PI) backbone and side poly(methyl methacrylate) chains (PI-PMMA) were prepared by mixing individual solutions of PA and PI-PMMA and subsequent film casting. Macromolecular brushes with the same backbone length and density of side chains but with various lengths of side PMMA chains were synthesized via activator generated by electron transfer atom transfer radical polymerization. Interactions between PA and PI-PMMA, as well as distribution of PI-PMMA filler inside the PA matrix, were studied by viscometry, dynamic light scattering, differential scanning calorimetry, scanning electron microscopy, and IR spectroscopy. The mechanical properties of polymer samples were also investigated. Microphase separation was revealed in PA/PI-PMMA films. The length of side chains influences interactions between PA and PI-PMMA. At the same time, the degree of reduction in the rigidity parameters and in the elasticity parameter depends on the side chain length. Diffusion membranes were prepared on the basis of compositions with the best mechanical properties; these membranes proved to be highly efficient in pervaporation of methanol–hexane mixture. It was established that the fluxes and separation factors of the studied membranes are several times higher as compared to the corresponding characteristics of known commercial membranes. POLYM. ENG. SCI., 60:481–490, 2020. © 2019 Society of Plastics Engineers     

Interferon-β Activity Is Affected by S100B Protein

Interferon-β (IFN-β) is a pleiotropic cytokine secreted in response to various pathological conditions and is clinically used for therapy of multiple sclerosis. Its application for treatment of cancer, infections and pulmonary diseases is limited by incomplete understanding of regulatory mechanisms of its functioning. Recently, we reported that IFN-β activity is affected by interactions with S100A1, S100A4, S100A6, and S100P proteins, which are members of the S100 protein family of multifunctional Ca²⁺-binding proteins possessing cytokine-like activities (Int J Mol Sci. 2020;21(24):9473). Here we show that IFN-β interacts with one more representative of the S100 protein family, the S100B protein, involved in numerous oncological and neurological diseases. The use of chemical crosslinking, intrinsic fluorescence, and surface plasmon resonance spectroscopy revealed IFN-β binding to Ca²⁺-loaded dimeric and monomeric forms of the S100B protein. Calcium depletion blocks the S100B–IFN-β interaction. S100B monomerization increases its affinity to IFN-β by 2.7 orders of magnitude (equilibrium dissociation constant of the complex reaches 47 pM). Crystal violet assay demonstrated that combined application of IFN-β and S100B (5–25 nM) eliminates their inhibitory effects on MCF-7 cell viability. Bioinformatics analysis showed that the direct modulation of IFN-β activity by the S100B protein described here could be relevant to progression of multiple oncological and neurological diseases.     

Simplified scoring methods for HMM-based speech recognition

Most of the contemporary speech recognition systems exploit complex algorithms based on Hidden Markov Models (HMMs) to achieve high accuracy. However, in some cases rich computational resources are not available, and even isolated words recognition becomes challenging task. In this paper, we present two ways to simplify scoring in HMM-based speech recognition in order to reduce its computational complexity. We focus on core HMM procedure—forward algorithm, which is used to find the probability of generating observation sequence by given HMM, applying methods of dynamic programming. All proposed approaches were tested on Russian words recognition and the results were compared with those demonstrated by conventional forward algorithm.     

Thermal treatment effect on structure, electrical conductivity and transient photoconductivity behavior of thiourea modified TiO2 sol-gel thin films

Thiourea modified nanocrystalline titanium dioxide (TiO₂) thin films were prepared by sol-gel route and were thermally treated at five different temperatures (400, 500, 600, 800 and 1000 °C). The films were studied using GIXRD, PIGE and UV-vis spectroscopy. It was observed that the anatase to rutile phase transformation of TiO₂ was inhibited by the thiourea modification. The transmittance of the modified films appeared reduced which was attributed both to the modification of TiO₂ with thiourea and the light scattering in the films. The dark conductivity and the transient photoconductivity of the modified TiO₂ sol-gel thin films were studied in vacuum and in air. The environment does not influence significantly the dark conductivity, because of the almost equivalent competition between oxygen and water adsorption. The photoconductivity reaches high values for all samples in both environments, with the sample treated at 500 °C to present the highest value. The larger values in vacuum can be attributed to the reduced amount of adsorbed oxygen at the surface, which acts as electron scavenger.     

Some Characteristic Features of Formation of Ceramics Based on Composite Powders SiC ― Si3N4 ― Si2N2O

We have studied the characteristic features of synthesis of composite powders SiC Si₃N₄ Si₂N₂O. We have investigated processes involving hot pressing of these powders without activating additives and a protective atmosphere. We consider the mechanical properties of ceramics obtained on the basis of these composite powders.     

Oxidative dehydrogenation of ethylbenzene to styrene over ultra-dispersed diamond and onion-like carbon

The catalytic properties of sp³-hybridized ultra-dispersed diamond and sp²-hybridized onion-like carbon in the oxidative dehydrogenation of ethylbenzene to styrene were investigated, highlighting the structure sensitivity of the reaction. The sp³-carbon led initially to C-C cleavage and benzene formation, while a switchover of the main reaction pathway into the styrene formation occurred with time on stream due to the formation of surface sp² carbon, required for the selective styrene formation. This was confirmed by the behavior and the high stable styrene selectivity shown by onion-like carbons. High temperature oxygen pre-treatment created catalytically active species at the sp² carbon surface, confirming that a high thermal stability carbon-oxygen complex was the active surface site for forming styrene.     

Some Characteristics of Free Cell Population in the Airways of Rats after Intratracheal Instillation of Copper-Containing Nano-Scale Particles

We used stable water suspensions of copper oxide particles with mean diameter 20 nm and of particles containing copper oxide and element copper with mean diameter 340 nm to assess the pulmonary phagocytosis response of rats to a single intratracheal instillation of these suspensions using optical, transmission electron, and semi-contact atomic force microscopy and biochemical indices measured in the bronchoalveolar lavage fluid. Although both nano and submicron ultrafine particles were adversely bioactive, the former were found to be more toxic for lungs as compared with the latter while evoking more pronounced defense recruitment of alveolar macrophages and especially of neutrophil leukocytes and more active phagocytosis. Based on our results and literature data, we consider both copper solubilization and direct contact with cellular organelles (mainly, mitochondria) of persistent particles internalized by phagocytes as probable mechanisms of their cytotoxicity.