Functional Characterization of Protein Kinase MAC-V with the Use of Two-Hybrid Cloning in Yeast

Identification of interaction partners opens a way to direct functional characterization of proteins. Several cDNAs coding for potential partners of protein kinase MAK-V/Hunk were isolated using two-hybrid cloning in yeast. Based on the partner properties, MAK-V/Hunk was assumed to play a role in tumorigenesis and tumor progression. With the previous results of two-hybrid cloning, MAK-V/Hunk was shown to participate in vesicular transport.     

The cardiac electric field in psychrophilic and thermophilic fish during atrial depolarization

In psychrophilic and thermophilic fish, significant differences were revealed under an optimal body temperature in the heart rate, initial atrial activity duration and descending slope of the P _II wave. Differences were also detected in atrial electric activity during the initial and final periods of depolarization, reflecting different localization of the initial excitation area and the movement of the depolarization front toward the atrioventricular border.     

The spectrum of mutations in genes associated with resistance to rifampicin,isoniazid, and fluoroquinolones in the clinical strains of <Emphasis Type="Italic">M. tuberculosis</Emphasis> reflects the transmissibility of mutant clones

To study the transmissibility of drug resistant mutant clones, M. tuberculosis samples were isolated from the patients of the clinical department and the polyclinic of the Central TB Research Institute (n = 1455) for 2011–2014. A number of clones were phenotypically resistant to rifampicin (n = 829), isoniazid (n = 968), and fluoroquinolones (n = 220). We have detected 21 resistance-associated variants in eight codons of rpoB, six variants in three codons of katG, three variants in two positions of inhA, four variants in four positions of ahpC, and nine variants in five codons of gyrA, which were represented in the analyzed samples with varied frequencies. Most common mutations were rpoB 531 Ser→Leu (77.93%), katG 315 (Ser→Thr) (94.11%), and gyrA 94 (Asp→Gly) (45.45%). We found that the mutations at position 15 of inhA (C→T) (frequency of 25.72%) are commonly associated with katG 315 (Ser→Thr). This association of two DNA variants may arise due to the double selection by coexposure of M. tuberculosis to isoniazid and ethionamide. The high transmissibility of mutated strains was observed, which may be explained by the minimal influence of the resistance determinants on strain viability. The high transmissibility of resistant variants may also explain the large populational prevalence of drug-resistant TB strains.     

Formulation of organic and inorganic hydrogel matrices for immobilization of β‐glucosidase in microfluidic platform

The aim of this study was to formulate silica and alginate hydrogels for immobilization of β‐glucosidase. For this purpose, enzyme kinetics in hydrogels were determined, activity of immobilized enzymes was compared with that of free enzyme, and structures of silica and alginate hydrogels were characterized in terms of surface area and pore size. The addition of polyethylene oxide improved the mechanical strength of the silica gels and 68% of the initial activity of the enzyme was preserved after immobilizing into tetraethyl orthosilicate–polyethylene oxide matrix where the relative activity in alginate beads was 87%. The immobilized β‐glucosidase was loaded into glass–silicon–glass microreactors and catalysis of 4‐nitrophenyl β‐d ‐glucopyranoside was carried out at various retention times (5, 10, and 15 min) to compare the performance of silica and alginate hydrogels as immobilization matrices. The results indicated that alginate hydrogels exhibited slightly better properties than silica, which can be utilized for biocatalysis in microfluidic platforms.     

Manifestation of the size effect during crystallization and melting of dispersed water in native and amorphous starches with various degrees of hydration

Differential scanning calorimetry (DSC) was used to study the melting and crystallization of frozen water dispersed in humid potato starch. Melting and crystallization temperatures and heats as functions of the degree of hydration of the starch were obtained for native and amorphous starch states. Manifestations of the size effect were observed in the dependences of heat for the processes in both starch states. Crystallization and melting heats of frozen water were found to change nonlinearly with the increasing degree of hydration in all cases. In contrast, a size effect in the dependences of melting and crystallization temperatures of frozen water was detected only for native starch. Reasons responsible for the absence of a size effect in the amorphous state were considered. Hysteresis, which is characteristic of small particles, was observed upon melting and crystallization of frozen water and its manifestation strongly differed in the native and amorphous states of potato starch.     

Reorganization of interphase microtubules in root cells of <Emphasis Type="Italic">Medicago sativa</Emphasis> L. during acclimation to osmotic and salt stress

We examined the organization of microtubule system of interphase cells in roots of Medicago sativa L. during acclimation to salt and osmotic stress at different concentrations of NaCl, Na₂SO₄, and mannitol. We identified morphological changes of tubulin cytoskeleton in different root tissues during the acclimation to salt and osmotic stress: (1) decreased density of the cortical microtubule network, (2) random orientation of cortical microtubule bundles, (4) thickening of the bundles, (3) nonuniform density of the bundles, (4) fragmentation of the bundles, and (5) formation of microtubule converging centers. Network thinning and thickening of the bundles were observed both under osmotic and salt stress. Random orientation of cortical microtubules was visualized under osmotic stress but not during salt stress. Fragmentation of microtubule bundles took place under salt stress with a high concentration of mannitol. Formation of microtubule converging centers was common under prolonged action of sodium sulfate, less evident under sodium chloride, and not found after mannitol treatment. Our data show that, in alfalfa root cells, cortical microtubules rearrange not only in response to different ions, but also to osmotic pressure. Thus, the signaling pathways and molecular mechanisms inducing reorganization of the microtubule system may be triggered by sodium cations, as well as by sulfate and chloride anions at concentrations that do not cause irreversible cell damage.     

Comparison of different pretreatment methods for lignocellulosic materials. Part I: conversion of rye straw to valuable products

The conversion of lignocellulose to valuable products requires I: a fractionation of the major components hemicellulose, cellulose, and lignin, II: an efficient method to process these components to higher valued products. The present work compares liquid hot water (LHW) pretreatment to the soda pulping process and to the ethanol organosolv pretreatment using rye straw as a single lignocellulosic material. The organosolv pretreated rye straw was shown to require the lowest enzyme loading in order to achieve a complete saccharification of cellulose to glucose. At biomass loadings of up to 15% (w/w) cellulose conversion of LHW and organosolv pretreated lignocellulose was found to be almost equal. The soda pulping process shows lower carbohydrate and lignin recoveries compared to the other two processes. In combination with a detailed analysis of the different lignins obtained from the three pretreatment methods, this work gives an overview of the potential products from different pretreatment processes.     

Polysaccharides of Algae: 55. Polysaccharide Composition of Several Brown Algae from Kamchatka

A new spectrophotometric procedure was developed for the quantitative determination of fucoidan and alginic acid and used for their analysis in extracts from biomass of 17 species of brown algae collected from coastal waters of the Kamchatka peninsula. In addition, neutral monosaccharides and mannitol were determined in the hydrolysis products of the alga biomass samples. The polysaccharide composition was shown to substantially depend on the algal species. The alginic acid content was maximal in the Alaria marginatablades; all the other representatives of the order Laminariales also are useful sources of this polysaccharide. At the same time, the fucoidan content is rather low in Laminariales. The highest content of fucoidan was found in Saundersella simplex, but Chordaria flagelliformisand Fucus evanescensare more practical fucoidan sources; the available supplies and the sugar composition make the latter alga the most suitable for the complex processing to prepare mannitol, fucoidan, and alginic acid.     

Composite materials based on chitosan and montmorillonite: prospects for use as a matrix for stem and regenerative cell cultivation

This paper examines the structural and mechanical properties of composite materials based on chitosan and micro- and nanoparticles of Na-montmorillonite and possibility of application for cultivation and targeted delivery of mesenchymal stem cells and regenerative cells. It's have been shown by addition of Na-montmorillonite biomaterial acquires stability of structural and mechanical properties in the sterilization process the handling of liquid media in cell culture. In vitro studies using dermal fibroblasts and adipose tissue mesenchymal stem cells demonstrated that this material has a set of properties to ensure matrix biocompatibility.