Role of thiol redox systems in <Emphasis Type="Italic">Escherichia coli</Emphasis> response to thermal and antibiotic stresses

Isogenic knockout mutants of Escherichia coli deficient in components of the glutathione and thioredoxin redox systems and growing at various temperatures (20–46°C) exhibited considerable differences in growth rate and survival, as well as in expression of the antioxidant genes. In the parental strain E. coli BW25113 (wt) treated with ciprofloxacin, ampicillin, or streptomycin, dependence of survival on growth temperature was a V-shaped curve with the maximum sensitivity within the range corresponding to high growth rates (40–44°C). Significant inverse correlation was observed between log CFU at different temperatures and specific growth rate prior to antibiotic addition. This applied to most of the mutants, which exhibited higher resistance to the three antibiotics tested at nonoptimal temperatures (20 and 46°C) than at 37 and 40°C. No correlation was found between resistance to stress and antibiotics and expression of the antioxidant genes. The role of global regulators ppGpp and σ^s in E. coli resistance to antibiotics and nonoptimal temperatures was shown.     

Increasing <Emphasis Type="Italic">Triticum aestivum</Emphasis> tolerance to cadmium stress through endophytic strains of <Emphasis Type="Italic">Bacillus subtilis</Emphasis>

Impact of inoculation of wheat seeds with endophytic strains of B. subtilis bacterium on revealing cadmium phytotoxicity of the plants was investigated. It was shown that, in the presence of Cd in the plants whose seeds were inoculated with the above bacteria, the activities of catalase and peroxidase and the content of nonprotein thiols were increased, while an intensity of lipid peroxidation decreased. Moreover, inoculation of plant seeds with the bacteria contributed to lowering the metal content in plant shoots.     

Hepatoprotective Activity of Betulin and Dipterocarpol Derivatives

Russian Journal of Bioorganic Chemistry - The regioselective synthesis of betulin 3,28-bis- and 28-monoacylates (nicotinate and isonicotinates) as well as of 3β-hemisuccinyl dipterocarpol was...     

Assessment of Effects of the OPRD1 and OPRM1 Genes Encoding Opioid Receptors on Apathy in Schizophrenia

Russian Journal of Genetics - Because of the involvement in motivational processes, opioid receptors are potential targets for apathy treatment in schizophrenia. We therefore searched for...     

Local polarity and hydrogen bonding inside the Sec14p phospholipid-binding cavity: high-field multi-frequency electron paramagnetic resonance studies

Sec14p promotes the energy-independent transfer of either phosphatidylinositol (PtdIns) or phosphatidylcholine (PtdCho) between lipid bilayers in vitro and represents the major PtdIns/PtdCho transfer protein in the budding yeast Saccharomyces cerevisiae. Herein, we employ multi-frequency high-field electron paramagnetic resonance (EPR) to analyze the electrostatic and hydrogen-bonding microenvironments for series of doxyl-labeled PtdCho molecules bound by Sec14p in a soluble protein-PtdCho complex. A structurally similar compound, 5-doxyl stearic acid dissolved in a series of solvents, was used for experimental calibration. The experiments yielded two-component rigid limit 130- and 220-GHz EPR spectra with excellent resolution in the gx region. Those components were assigned to hydrogen-bonded and nonhydrogen-bonded nitroxide species. Partially resolved 130-GHz EPR spectra from n-doxyl-PtdCho bound to Sec14p were analyzed using this two-component model and allowed quantification of two parameters. First, the fraction of hydrogen-bonded nitroxide species for each n-doxyl-PtdCho was calculated. Second, the proticity profile along the phospholipid-binding cavity of Sec14p was characterized. The data suggest the polarity gradient inside the Sec14p cavity is a significant contributor to the driving molecular forces for extracting a phospholipid from the bilayer. Finally, the enhanced g-factor resolution of EPR at 130 and 220 GHz provides researchers with a spectroscopic tool to deconvolute two major contributions to the x-component of the nitroxide g-matrix: hydrogen-bond formation and local electrostatic effects.     

Discrimination of heterozygous carriage of ataxia-telangiectasia by the method of indirect immunofluorescent analysis

Qualitative differences of the stabilization of protein P53 were studied by methods of indirect immunofluorescence with the use of confocal microscopy in cells obtained from members of the family of a patient with ataxia-telangiectasia (AT)—the proband AT6SP and her blood relatives: the sister AT(S)6SP and the father AT(F)6SP—at different time periods after irradiation with various doses of ionizing radiation.     

Differential expression of duplicated LDH-A genes during temperature acclimation of weatherfish Misgurnus fossilis. Functional consequences for the enzyme

Temperature acclimation in poikilotherms entails metabolic rearrangements provided by variations in enzyme properties. However, in most cases the underlying molecular mechanisms that result in structural changes in the enzymes are obscure. This study reports that acclimation to low (5 degrees C) and high (18 degrees C) temperatures leads to differential expression of alternative forms of the LDH-A gene in white skeletal muscle of weatherfish, Misgurnus fossilis. Two isoforms of LDH-A mRNA were isolated and characterized: a short isoform (= 1332 bp) and a long isoform ( = 1550 bp), which both have 5'-UTRs and ORFs of the same length (333 amino acid residues), but differ in the length of the 3'-UTR. In addition, these two mRNAs have 44 nucleotide point mismatches of an irregular pattern along the complete sequence, resulting in three amino acid mismatches (Gly214Val; Val304Ile and Asp312Glu) between protein products from the short and long mRNA forms, correspondingly LDH-A(alpha) and LDH-A(beta) subunits. It is expected that the beta-subunit is more aliphatic due to the properties of the mismatched amino acids and therefore sterically more restricted. According to molecular modelling of M. fossilis LDH-A, the Val304Ile mismatch is located in the subunit contact area of the tetramer, whereas the remaining two mismatches surround the contact area; this is expected to manifest in the kinetic and thermodynamic properties of the assembled tetramer. In warm-acclimated fish the relative expression between alpha and beta isoforms of the LDH-A mRNA is around 5 : 1, whereas in cold-acclimated fish expression of is reduced almost to zero. This indicates that at low temperature the pool of total tetrameric LDH-A is more homogeneous in terms of alpha/beta-subunit composition. The temperature acclimation pattern of proportional pooling of subunits with different kinetic and thermodynamic properties of the tetrameric enzyme may result in fine-tuning of the properties of skeletal LDH-A, which is in line with previously observed kinetic and thermodynamic differences between 'cold' and 'warm' LDH-A purified from weatherfish. Also, an irregular pattern of nucleotide mismatches indicates that these mRNAs are the products of two independently evolving genes, i.e. paralogues. Karyotype analysis has confirmed that the experimental population of M. fossilis is tetraploid (2n = 100), therefore gene duplication, possibly through tetraploidy, may contribute to the adaptability towards temperature variation.     

Effect of Low-Frequency Pulsed Magnetic Field and Low-Level Laser Radiation on Oxidoreductase Activity and Growth of Fungi—Active Destructors of Polymer Materials

Microbiology - Effect of low-frequency pulsed magnetic field and of low-intensity laser radiation on mycelial fungi actively degrading various polymer materials was studied. These factors had a...     

Excess of rare amino acid polymorphisms in the Toll-like receptor 4 in humans

The Toll-like receptor 4 protein acts as the transducing subunit of the lipopolysaccharide receptor complex and assists in the detection of Gram-negative pathogens within the mammalian host. Several lines of evidence support the view that variation at the TLR4 locus may alter host susceptibility to Gram-negative infection or the outcome of infection. Here, we surveyed TLR4 sequence variation in the complete coding region (2.4 kb) in 348 individuals from several population samples; in addition, a subset of the individuals was surveyed at 1.1 kb of intronic sequence. More than 90% of the chromosomes examined encoded the same structural isoform of TLR4, while the rest harbored 12 rare amino acid variants. Conversely, the variants at silent sites (intronic and synonymous positions) occur at both low and high frequencies and are consistent with a neutral model of mutation and random drift. The spectrum of allele frequencies for amino acid variants shows a significant skew toward lower frequencies relative to both the neutral model and the pattern observed at linked silent sites. This is consistent with the hypothesis that weak purifying selection acted on TLR4 and that most mutations affecting TLR4 protein structure have at least mildly deleterious phenotypic effects. These results may imply that genetic variants contributing to disease susceptibility occur at low frequencies in the population and suggest strategies for optimizing the design of disease-mapping studies.     

Interlineage distribution and characteristics of the structure of two subfamilies of Drosophila melanogaster MDG4 (gypsy) retrotransposon

The distribution of two variants of MDG4 (gypsy) was analyzed in several Drosophila melanogaster strains. Southern blot hybridization revealed the inactive variant of MDG4 in all strains examined and active MDG4 only in some of them. Most of the strains harboring the active MDG4 variant were recently isolated from natural populations. It is of interest that the active MDG4 prevailed over the inactive one only in strains carrying the mutant flamenco gene. Several lines were analyzed in more detail. The number of MDG4 sites on salivary-gland polytene chromosomes was established via in situ hybridization, and MDG4 was tested for transposition using the ovoD test.