[STUDY OF COMPOSITION OF PLANT EXTRACTS, POSSESSING ANTIMICROBIAL EFFECT AGAINST VIBRIO CHOLERAE EL TOR, USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY].

AIM: Study the composition of plant extracts using high-performance liquid chromatography. (HPLC) and evaluation of their antimicrobial effect against Vibrio cholerae El Tor. MATERIALS AND METHODS: Qualitative and quantitative composition of plant extracts was studied using HPLC. Determination of sensitivity of microorganisms to plant extracts was carried out by diffusion into agar method and serial dilutions method. RESULTS: Antibacterial effect of water, water-alcohol and acetone extracts of roots of Limonium gmelinii L., Berberis vulgaris L. and Glycyrrhiza glabra L. was studied. The most effective methods of extraction of biologically active substances, possessing antimicrobial effect against various strains of V. cholerae El Tor, were determined. CONCLUSION: The use of HPLC allowed to establish the presence of catechines, alkaloids protoberberines and glycyrrhizic acid in xtracts, possessing antimicrobial effect against V. cholera El Tor strains.

[Effect of plant extracts on cytotoxic activity of Vibrio cholerae hemolysin].

AIM: Study of plant extracts that have the ability to neutralize cytotoxic activity of hemolysin. MATERIALS AND METHODS: Preparations of purified and recombinant V. cholerae eltor hemolysin as well as supernatants of V. cholerae strains were used. Determination ofcytotoxic activity of hemolysin and neutralizing activity of plant extracts were carried out by using cell cultures CHO-K1 and CaCo2. RESULTS: Out of 9 water extracts only 3 - extracts of Rhei rhizome, Limonium gmelinii and Quercus robur neutralized hemolysin in cell culture CHO-K1 and CaCo2, whereas the other extracts--Humulus lupulus, Ocimum basilicum, Chelidonium majus, Juglans regia, Achillea milefolium and Hypericum perforatum did not have anti-cytotoxic effect. Neutralizing properties of extracts are exhibited during their co-incubation with hemolysin preparations and supernatants of V. cholerae strains already within 10 minutes. CONCLUSION: Plant extracts that have anti-cytotoxic activity against hemolysin are perspective for development oftherapeutic-prophylaxis preparations.

[Influence of plant extracts on the activity of cholera toxin of Vibrio cholerae].

AIM: Study the activity of plant extracts against cholera toxin (CT) of Vibrio cholerae O1. MATERIALS AND METHODS: Antitoxic activity of plant extracts was determined by using enzyme immunoassay and CHO-K1 cell culture. RESULTS: 8 water extracts of plants were studied. Extracts of nut, tutsan, milfoil, basil do not have effect on CT activity in EIA or CHO-K1 cell culture. Celandine and rhubarb extracts do not reduce CT immunochemical activity but prevent elongation of CHO-K1 cells. Oak and hop extracts suppress binding in EIA of cholera toxin and GM1 receptors and insignificantly reduce its activity in cell culture. CONCLUSION: Antitoxic activityofplant extracts against CT is perspective for the development of preparations possessing inhibition effect.

[GM1-dot-EIA for the detection of toxin-producing Vibrio cholerae strains].

A new variant of enzyme immunoassay (EIA) has been developed on the basis of GM1 gangliosides to detect the toxin-producing Vibrio cholerae strains--GM1-dot-EIA. Experiments were run using a nitrocellulose membrane to bind GM1 gangliosides and polyclonal antitoxic serum to detect cholerogen. GM1-dot-EIA testing identified cholera toxin in 11 of 13 supernatants of V. cholerae eltor ctx(+) strains isolated from man and in 3 of 7 supernatants of V. cholerae eltor ctx(+) strains isolated from water. These data agree with those obtained in CM1-EIA. There was no reaction with the supernatants of other microorganisms. The sensitivity of the technique was 10 ng/ml. Thus, the simple and specific GM1-dot-EIA may be recommended to detect toxin-producing V cholerae strains isolated from man and water.

[Heat shock modification of nitrosomethylurea action on sunflower seedlings]. / Modifikatsiia deistviia nitrozometilmocheviny na prorostki podsolnechnika teplovym shokom.

Separate and combined action of nitrosomethylurea (NMU) and heat shock (HS) on germination of seeds of the inbred line 3629 and derived nuclear and plastome mutant lines, n-chlorina-1 and en-chlorina-5, respectively, were studied. NMU at a concentration of 0.015%, as well as HS (40 degrees C), had no effect on the spontaneous level of chromosome aberrations in root meristem of sunflower seedlings. However, at a concentration of 0.03%, the mutagen considerably increased the frequency of chromosome rearrangements. Pretreatment with heat for 30 min enhanced the cytogenetic effect of NMU (0.03%) on line 3629 seedlings. In the nuclear mutant, the reaction of root meristematic cells did not depend on the additional heat treatment. The combined action of HS and NMU at early germination stages had no effect on the survival of line 3629 plants and plastome mutant en-chlorina-5; the survival of n-chlorina-1 plants decreased. The highest frequency of M1 plants with chlorophyllic abnormalities (green revertants, lethals, and variegated forms) was observed in n-chlorina-1 line.

[Mutagenic effect of nitrosomethylurea, modified by heat shock at early stages of sunflower seedling development]. / Mutagennyi éffekt nitrozometilmochevinyi, modifitsirovannyi teplovym shokom na rannikh étapakh razvitiia prorostkov podsolnechnika.

A comparative study of chlorophyll mutations induced in sunflower seedlings of the initial line 3629 and its nuclear and plastom mutant derivatives by nitrosomethylurea (NMU) (0.015%), heat shock (HS, 40 degrees C), and their combination was performed. The spectrum and frequency of chlorophyll mutations depended on the treatment conditions and on the genotype of the lines tested. Nuclear mutant displayed highest sensitivity to NMU, HS, and their combination. The line 3629 plants displayed highest tolerance to NMU and HS. However, plastom mutants were more tolerant to the combination of the mutagens compared to the plants of the initial line.