[Effect of Acholeplasma laidlawii var. granulum on lectin activity of sugar beet calluses].

The dynamics of lectin activity of sugar beet calluses infected by mollicute Acholeplasma laidlawii var granulum str. 118 was studied. Activity of acid-soluble lectins of sugar beet cell cultures increased 4.5 times during some first hours after infecting by acholeplasma. At early stages of infecting by mollicute qualitative changes were also revealed in the protein spectra of acid-soluble lectins of sugar beet calluses.

[The inhibition of Mycoplasma pneumoniae adhesion in a fetuin test system by synthetic analogs and polymeric forms of neuraminic acid]. / Ingibirovanie adgezii Mycoplasma pneumoniae v fetuinovoi test-sisteme sinteticheskimi analogami i polimernymi formami neiraminovoi kisloty.

Eight glycosides and structural analogues of neuraminic acid as well as eight polymeric forms of N-acetyl neuraminic acid have been studied for their inhibitory effect on adhesion of Mycoplasma pneumoniae. Maximum inhibiting effect among low-molecular compounds was manifested by 2-->3 sialyllactose which, being used in concentrations 5.0 and 10.0 micrograms/ml, inhibited adhesion of mycoplasmas by 76 and 87%, respectively. These indices for other derivatives in the above mentioned concentrations were as follows (%): 2-->6 sialyllactose, 31 and 74%; alpha-Me-glycoside NeuAc, 75 and 85%; alpha-Bn-glycoside-N-trifluoruracetyl NeuAc, 30 and 63%; alpha-Bn-glycoside NeuAc, 32 and 59%; alpha-Bn-glycoside-4-epi-NeuAc, 20 and 27%; beta-Bn-glycoside NeuAc, 2-4%; beta-me-glycoside NeuAc, 4-5%. The maximum inhibiting effect (50% inhibition at concentration 2.5 mumol) among polymeric forms was exerted by the conjugate alpha-benzeneglycoside with polyacrylic acid containing 12 mol% of NeuAc. Conjugates with 8, 16 and 20 mol% of NeuAc possessed a bit less activity. The 50% concentration for them was 5.3, 3.1 and 8.3 mumol, respectively. Polymeric forms on the basis of polyacrylamide proved less active.

[Mono- and polyvalent synthetic sialosides as inhibitors of Mycoplasma pneumoniae adhesiveness]. / Mono- i polivalentnye sinteticheskie sialozidy kak ingibitory adgezivnosti Mycoplasma pneumoniae.

Nine mono- and polyvalent sialosides as substances inhibiting adhesive properties of mycoplasma Mycoplasma pneumoniae--the agent of human atypical pneumonia have been studied in the reaction of hemagglutination (RHA) using solid-phase variant of ELISA-test ("sandwich"--variant), which is based on the competition for specific binding in RHA between the studied syalosides and sialylglycoproteins of fetuin conjugate with horseradish peroxidase. Of seven polymers--P alpha.12.ONa(N1), P alpha.12.EA(N2), P alpha.12.NH2(N3), P alpha.12.AES.10.ONa(N4), P alpha.12.Hg.20.ONa(N6), P20.SLea.NH2(N7), as well as monomer alpha-Bn Neu5Ac and fucoidan the polymers 5, 2, 4 and 7 in concentrations as to the content of sialic acid 1.0; 1.3; 1.35 and 10.0 M, respectively, most efficiently (up to 98%) inhibited the adhesiveness of M. pneumoniae. Polymeric sialosides 3, 6 and 1 proved less active and, the concentration of sialic acid in the composition of their molecules being 10.0 microM, inhibited adhesiveness of M. pneumoniae by approximately 77, 75 and 62.5%, respectively. Antiadhesive activity of fucoidan and monomer proved too low under concentration of these substances as to the content of sialic acid in them 25 microM: they decreased the ability to adhesion in M. pneumoniae by 33 and 56%, respectively. This proved that polyvalent sialosides 2, 4, 5 and 7 are promising for creation of drugs for treatment and prophylaxis of human pneumonias of mycoplasmal etiology on principally new basis with regard for the properties of the disease agent.

[The isolation and partial purification of 2 DNA-dependent DNA polymerases from Acholeplasma laidlawii PG-8]. / Vydelenie i chastichnaia ochistka dvukh DNK-zavisimykh DNK-polimeraz Acholeplasma laidlawii PG-8.

Two forms of DNA-dependent DNA-polymerase have been isolated and partially purified from the limited amount of biomass of cells Acholeplasma laidlawii PG-8, a typical representative of genus Acholeplasmataceae, as a result of successive chromatography on the columns with DEAE-cellulose DE-52 and Green A-sepharose. The first form of DNA-polymerase is eluted from the ion-exchange column with NaCl concentration of 0.1 M from the column with Green A-sepharose of 0.27 M, while the second form-with NaCl concentrations of 0.6 and 0.4 M, respectively. The both enzymatic activities are able to implement DNA synthesis. The conditions of DNA-polymerase production proved to be rather convenient for isolation of the concentrated and highly active enzymes.

[A trial of cultivating Rickettsiella phytoseiuli on the SM IMV-72 medium used for growing phytopathogenic mycoplasmas]. / Popytka kul'tivirovaniia Rickettsiella phytoseiuli v srede SM IMV-72, primeniaemoi dlia vyrashchivaniia fitopatogennykh mikoplazm.

The work presents data on growing Rickettsiella phytoseiuli in the SM IMV-72 medium which is used for cultivation of phytopathogenic mycoplasmas. Rickettsiella was observed till the 66th day in the primary cultures and till the 16th day after the first passage. Binary division of the cells has been found only in the primary cultures; a complex cycle of the Rickettsiella development ceased at the stage of formation of the crystal-forming cells. Attempts to passivate Rickettsiella were failure.

[The effect of physicochemical factors on the activity of DNA-dependent DNA polymerases in Acholeplasma laidlawii PG-8]. / Vliianie nekotorykh fiziko-khimicheskikh faktorov na aktivnost' DNK-zavisimykh DNK-polimeraz Acholeplasma laidlawii PG-8.

The biological and physico-chemical properties of DNA-dependent DNA-polymerases of Acholeplasma laidlawii PG-8 have been studied. The optimal parameters of maximal enzymatic activity are determined. It is stated that N-ethylmaleimide in concentration of 1 mM activated DNA-polymerase I by 52%, whereas DNA-polymerase II with reagent concentration of 0.5 mM demonstrated the peak of activity exceeding the control only by 10%. Spermidine in concentration of 1.5 mM for the first form of DNA-polymerase and 0.15 mM-for the second one increased the ability of both forms of polymerases to synthesize DNA by 10%. Aphidicolin added to the reaction medium up to concentration of 10 mg/ml decreased activity of forms I and II of enzymes by 83 and 68%, respectively. The presence of 0.6 mM of EDTA in the medium also negatively affected the activity of polymerases inhibiting it by 83% in form I and by 77%-in form II.

[Choice of the optimal method of numerical analysis in taxonometric study of mollicutes based on electrophoresis of their proteins]. / Vybor optimal'nogo metoda numericheskogo analiza dlia taksonomicheskogizucheniia mollikutov na osnove élektroforeza ikh belkov.

Different procedures of numerical analysis are evaluated to select the most convenient of them for processing the results of mycoplasm protein electrophoresis in polyacrylamide gel (EPAAG) when solving problems of their taxonomy and systematics. This aim is satisfied most of all by the method of Sokal which permits revealing relations at the rank of species, genus and higher taxons. This method as well as the approaches developed by the authors for transforming results of EPAAG of proteins into the numerical form are recommended to improve the systematics of mollicutes with application of computers.

[The inhibitory action of 6-azacytidine on Mollicutes and its proposed mechanism]. / Ingibitornoe deistvie 6-azatsitidina na mollikuty i ego predpolagaemyi mekhanizm.

6-Azacytidine (6-AC) is shown to have an inhibitory effect on the Mollicutes of the different systematic position. The growth of type strains of Mollicutes (Acholeplasma laidlawii PG-8, Mycoplasma pneumoniae FH and M. fermentans PG-18) completely ceased in the nutrient medium at concentration of the above substance in it within the range of 125-250 micrograms/ml. 50% inhibiting concentration of 6-AC equaled: for M. fermentans PG-8: 23.43 micrograms/ml; M. pneumoniae FN: 46.8 micrograms/ml; Acholeplasma laidlawii PG-8: 62.5 micrograms/ml. 6-AC concentration 5 micrograms/ml decreased the process of DNA-dependent DNA synthesis in the in vitro system more than by 60%. 6-AC exerted less effect on the DNA-dependent RNA synthesis in the in vitro system: at different concentrations of 6-AC (up to 400 micrograms/ml) RNA synthesis decreased only by 20%. Translation on ribosomes of Mollicutes in the in vitro system completely ceased at 6-AC concentration 100 micrograms/ml. The results obtained indicate that for 6-AC in cells of Mollicutes and, possibly, for other microorganisms there are two targets: ribosomes and DNA-dependent DNA-polymerase. Total effect of blocking of the translation and replication processes by 6-azacytidine causes death of Mollicutes. Since 6-AC has no harmful effect on the human cells, it can be used as an efficient method for treatment of respiratory and urogenital diseases induced by Mollicutes.