Increased secretion of lignolytic enzymes by the Lentinus tigrinus fungus after addition of butanol and toluene in submerged cultivation

We studied the effects of butanol and toluene on secretion of lignolytic enzymes by the Lentinus tigrinus fungus during submerged cultivation. Addition of butanol and toluene during the trophophase was followed by an increase in laccase and peroxidase activity of the culture and change in the composition of phospholipids and fatty acids. The ratio of phosphatidylcholine and phosphatidic acid decreased, while the amount of lysophosphatidylcholine, phosphatidylethanolamine, phosphoinositides, phosphatidylserine, and unsaturated fatty acids decreased. These changes resulted in an increase in the unsaturation index.     

Study of NO-synthase activity and detection of NO-dependent antimicrobial effects in primary culture of frog urinary bladder epithelial cells

We have shown previously that endogenous nitric oxide (NO) in frog urinary bladders modulates the influence of arginine-vasotocin on the increase in the osmotic water permeability of the bladder epithelium. The goal of the present work was to obtain a primary culture of epithelial cells from a frog urinary bladder in order to study in vitro the cellular activity of NO-synthase (NOS) and its regulation. The best conditions for cultivating the cells turned out to be with the use of a modified L-15 medium containing 10% fetal bovine serum and gentamicin (40 μg/ml) at room temperature. Under these conditions, at least 50% of the cells preserved their viability for 8 days. The NOS activity was estimated from the accumulation of nitrites (NO ₂ ^? ) in the cultivation medium; the amount of NO ₂ ^? in the presence of nitro-L-arginine methyl ester (L-NAME), an inhibitor of all types of NOS, was considered unspecific and was subtracted from each value. The NO ₂ ^? accumulation was linear in time for three days of cultivation and was inhibited by 1400W, an inducible NOS (iNOS) inhibitor, or by 7-nitroindazole, an inhibitor of constitutive NOSs, at concentrations of 5–50 and 10–200 μM, respectively. One-day incubation of the cells in the medium with low doses of gentamicin (1 or 2 μg/ml) led to a marked increase in the amount of bacteria in the cultivation m5 mM edium identified as E. coliand Acinetobacter sp. An addition of 5 mM L-NAME to the cultivation medium increased the amount of the bacteria by 1.5-and 2.5-fold in the presence of 2 and 1 μg/ml gentamicin/ml, respectively. Thus, the epithelial cells of frog urinary bladders have the NO-dependent antibacterial effect, which seems to be the result of enhanced iNOS expression. The obtained data indicate that the primary culture of the epithelial cells from frog urinary bladders is a prospective model for studying the role of NO and the regulation of NOS activity, which is of particular interest for studying protective NO effects in epithelial tissues.     

Electrical responses of <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> to light stimulation: Effect of divalent cations

The effects of Ca²⁺, Mg²⁺ and Mn²⁺ on light-evoked electrical responses, ERG and action potentials of optic nerve fibers, were studied in an isolated eye of Lymnaea stagnalis. Signals of both types persisted with minor changes when Mg²⁺ concentration in the bath solution was increased up to 15 mM. This finding suggests that action potentials of photoreceptor cells are transmitted via their axons directly to central ganglia without a relay in chemical synapses on interneurons. At the same time, the changed spiking pattern may indicate chemical interactions between photoreceptors cells themselves. In the presence of 4 mM EDTA or 10 mM Mn²⁺, both ERG waves and spiking activity were considerably depressed, probably, as a result of impaired phototransduction in photoreceptor cells.     

Crosslinking of Cys142 of Methyltransferase SsoII with DNA Duplexes Containing a Single Internucleotide Phosphoryldisulfide Link

DNA duplexes containing a single phosphoryldisulfide link in place of the natural internucleotide phosphodiester bond were employed in affinity modification of Cys142 in cytosine-C5 DNA methyltransferase SsoII (M.SsoII). The possibility of duplex–M.SsoII conjugation as a result of disulfide exchange was demonstrated. The crosslinking efficiency proved to depend on the DNA primary structure, modification position, and the presence of S-adenosyl-L-homocysteine, a nonreactive analog of the methylation cofactor. The SH group of M.SsoII Cys142 was assumed to be close to the DNA sugar-phosphate backbone in the DNA–enzyme complex.     

Effect of the chemical nature of the carbon source in the medium on the makeup of the biomass]

The yeast Candida tropicalis, Candida utilis, Trichosporon cutaneum were cultivated on the synthetic medium containing additions of group B vitamins (biotin, thiamin) and glucose, glycerol of lactic acid as the sole source. The protein content, amino acid composition, nucleic acids and complex B vitamins were identified in the resultant biomass. The carbon source in the medium affected these indices. The protein and nucleic acid content also depended on the yeast strain.     

Isolation of new species of the genus Actinomadura on selective media with streptomycin and bruneomycin]

In the screening programme using selective media with streptomycin (25, 50 and 100 gamma/ml) and bruneomycin (0.5, 1 and 2 gamma/ml) 3 unusual actinomycetous cultures forming short, rarely long chains of straight, hook-like or spiral spores with 1 or rarely 2 turns were isolated. The study of the cell wall composition of the isolates showed that the walls contained meso-diaminopimelic acid and a sugar madurose. On the basis of the morphological structure and the cell wall composition the isolates were classified as belonging to Actinomadura. Comparison of the cultures with the Actinomadura species described in the literature showed that the isolates were new species of the genera, i. e. Am. coerulea sp. nov., Am. cremea sp. nov. and Am. salmonea sp. nov.     

A comparative analysis of Stachybotrys chartarum strains isolated in Russia

This work deals with a comparative analysis of Stachybotrys chartarum strains isolated from various artificial cellulose-containing materials and natural substrates in the geographically distant regions of Russia. The analysis included the determination of the spore size, the strain toxicity to Paramecium caudatum, the strain resistance to the fungicides Benomil, Olilen, and Tilt, and the PCR study of the genome structure with the aid of a primer that was complementary to the core sequence of the SINE retrotransposon. It was found that some of the strains that were isolated from different areas and from different substrates differ in their toxicity, fungicide resistance, and genome structure. The PCR analysis showed the absence of any correlation between the genome structure, the strain properties, the geographic area, and the substrates from which the strains were isolated. The pheno- and genotypic diversity of the strains and their different vegetative compatibility suggest the existence of an intraspecies diversity of the S. chartarum strains that were isolated in different geographic areas. The absence of any correlation between the pheno- and genotypic properties of the strains and the substrates from which they were isolated implies that the colonization of artificial substrates by S. chartarum occurred occasionally from natural habitats. The S. chartarum populations that live on artificial substrates are unlikely to have their own evolutionary history.