Cloning and expression of the determinant for resistance to a new class of tetracycline in Escherichia coli strains

Tetracycline-resistance determinant of the plasmid pBS221 isolated from a Pseudomonas aeruginosa strain has been cloned on pUC19 vector plasmid. The determinant is expressed under aerobic and anaerobic conditions coding for two proteins: a 36 Kd protein conferring antibiotic resistance and 27 Kd repressor protein. The determinant is not homologous to tet-determinants of the known classes as shown by blot hybridization experiments. The determinant represents a new class--G. Determinants of the new class are widespread among Serratia marcescens strains.     

Modification of blood plasma lipoproteins by a lipid peroxidation product--hexanal

HDL treated with hexanal is shown to lose the ability for the cholesterol absorption. In the case of LDL at low concentration of the modifying agent the rate of their elimination from the blood stream of the rabbit decrease, but their uptake by the rat macrophages do not differ from the uptake of native lipoproteins. At high concentration of hexanal the rate of the elimination of LDL from the blood stream increases considerably and is close to that of acetylated LDL. Thus, the modification of plasma lipoproteins with monoaldehydes occurring in the aorta wall leads to the loss of the functional properties of the lipoprotein particles.     

Comparative Genomic Analysis of Mycobacterium tuberculosis Drug Resistant Strains from Russia

Tuberculosis caused by multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis (MTB) strains is a growing problem in many countries. The availability of the complete nucleotide sequences of several MTB genomes allows to use the comparative genomics as a tool to study the relationships of strains and differences in their evolutionary history including acquisition of drug-resistance. In our work, we sequenced three genomes of Russian MTB strains of different phenotypes – drug susceptible, MDR and XDR. Of them, MDR and XDR strains were collected in Tomsk (Siberia, Russia) during the local TB outbreak in 1998–1999 and belonged to rare KQ and KY families in accordance with IS6110 typing, which are considered endemic for Russia. Based on phylogenetic analysis, our isolates belonged to different genetic families, Beijing, Ural and LAM, which made the direct comparison of their genomes impossible. For this reason we performed their comparison in the broader context of all M. tuberculosis genomes available in GenBank. The list of unique individual non-synonymous SNPs for each sequenced isolate was formed by comparison with all SNPs detected within the same phylogenetic group. For further functional analysis, all proteins with unique SNPs were ascribed to 20 different functional classes based on Clusters of Orthologous Groups (COG). We have confirmed drug resistant status of our isolates that harbored almost all known drug-resistance associated mutations. Unique SNPs of an XDR isolate CTRI-4^XDR, belonging to a Beijing family were compared in more detail with SNPs of additional 14 Russian XDR strains of the same family. Only type specific mutations in genes of repair, replication and recombination system (COG category L) were found common within this group. Probably the other unique SNPs discovered in CTRI-4^XDR may have an important role in adaptation of this microorganism to its surrounding and in escape from antituberculosis drugs treatment.     

Spatial distribution and species composition of prosthecate bacteria in Lake Baĭkal

From the water column of Lake Baikal, several strains of prosthecate bacteria belonging to the genera Caulobacter and Brevundimonas were isolated. In this article, the methods applied for their isolation and cell number determination are described; the occurrence frequency and spatial distribution of these microorganisms in the lake are demonstrated. Characterization of the species composition of cultivable and uncultivable prosthecate bacteria was carried out using the methods of traditional and molecular microbiology, respectively. A comparative phylogenetic analysis of the DNA sequences of uncultivable bacteria, which showed homology to the members of the alpha subclass of proteobacteria, was carried out. It was demonstrated that the lake water column is inhabited by uncultivable alpha-proteobacteria of uncertain phylogenetic affinity, in addition to representatives of the species Caulobacter vibrioides and C. leidyi, which were detected by traditional microbiological methods.     

Antiviral activity of lambda-, beta-, and gamma-interferons in the presence of theophylline: involvement of 2',5'-oligo(A) synthetase

Theophylline, an inhibitor of cAMP phosphodiesterase, induces in human ovary carcinoma cells (CaOv) a 2-2.5-fold elevation of intracellular cAMP. This rise in the cAMP level is followed by an increase of the activity of 2',5'-oligo(A) synthetase in CaOv cells -insignificant (1.5-fold) after 16 hr incubation, and substantial (3.7-fold) after 30 hr incubation, as well as the development of antiviral resistance. Once CaOv cells have been incubated with the mixtures containing theophylline (2 mM) and lambda-, beta-, and gamma-interferon preparations (0.5-13 IU/ml), the total antiviral effect of the mixtures exceeds that generated by interferon or theophylline separately; the action of the above agents being additive. These data agree with the previously obtained results and support the suggestion that cAMP phosphodiesterase inhibitors partially mimic the antiviral action of interferon.     

Degradation of bis-(2-Ethylhexyl)phthalate by Microorganisms of the Water and Bottom Sediments of the Selenga River and Lake Baikal under Experimental Conditions

Degradation of bis-(2-ethylhexyl)phthalate (BEHP) by microbial associations of the water and bottom sediments of the Selenga River and Lake Baikal and by pure cultures of microbial species belonging to various taxa isolated from the sediments under discussion is studied. It is shown that intense biological degradation occurs in both water and sediments. The degrees of conversion in closed experimental systems on minimal media are 46 and 24%, respectively. The most active of the organisms studied is a Micromironospora actinomycete. It degraded BEHP by 36% of its initial concentration. Spore-forming bacteria and microorganisms of the genus Pseudomonas were less active (17–23% and 7–11%).     

The Biodiversity of Actinomycetes in Lake Baikal

The taxonomic analysis of 107 actinomycete strains isolated from the bottom sediments and water of Lake Baikal showed that most of the water isolates belong to the genus Streptomyces and most of the sediment isolates belong to the genus Micromonospora. In the sediments, the number of actinomycetes increased with depth (down to 200 m). Eight Streptomyces isolates were identified to a species level.     

Dependence of Malate Dehydrogenase Structure on the Type of Metabolism in Freshwater Filamentous Colorless Sulfur Bacteria of the Genus Beggiatoa

Major pathways of carbon metabolism were studied in strains D-402 and D-405 of freshwater colorless sulfur bacteria of the genus Beggiatoa grown organotrophically and mixotrophically. The bacteria were found to possess all the enzymes of the tricarboxylic acid (TCA) and glyoxylate cycles. When organotrophic growth changed to mixotrophic growth, the activity of the TCA cycle enzymes decreased 2- to 3-fold, but the activity of enzymes of the glyoxylate cycle increased threefold. It follows that, in the oxidation of thiosulfate, organic compounds no longer play the leading part in the energy metabolism, and most of electrons that enter the electron transport chain (ETC) derive from inorganic sulfur compounds. A connection was established between the structure and kinetic characteristics of malate dehydrogenase—an enzyme of the TCA and glyoxylate cycles—and the type of carbon metabolism in the strains studied. Malate dehydrogenase in organotrophically grown cells of strains D-402 and D-405 is dimeric, whereas in strain D-402 grown mixotrophically it is tetrameric.