[EFFECT OF MYCOPLASMA INFECTION TO FATTY ACID COMPOSITION OF CALLUS CULTURE SUGAR BEET].

It was studied the effect of Acholeplasma laidlawii var. granulum str. 118 to fatty acid composition of sugar beet calluses. It was established that acting of acholeplasma results to changes in the quantitative content of the individual fatty acids and in the qualitative composition of fatty acids in the lipids of calluses. The changing of the fatty acid composition of calluses lipids of sugar beet infected by A. laidlawii vargranulum str. 118 is observed as nonspecific response to biotic stress.

[Activity of oxidative enzymes in plant cells under conditions of experimental mycoplasmosis].

Changes in activity of oxidative enzymes was shown when infecting the callus culture of sugar beet cells by phytopathogenic mollicute Acholeplasma laidlawii var granulum 118. Maximum indices of the enzyme activity were fixed 3 hours after infecting with regard to control: it was 49% for peroxidase, 38% for catalase and 45% for polyfenoloxidase. The decrease of enzymatic activity to 22%, 12% and 19%, respectively, was registered 5 hours after infecting, and after that all these indices became stabilized. Changes in the activity of components of antioxidative protection were probably connected with induction of plants' protective mechanisms in response to penetration of pathogenic mollicutes.

[Effect of wheat germ agglutinin on acholeplasma--the pathogen of phytomycoplasmosis].

Introduction of wheat germ agglutinin (WGA) to cultural medium of pale green dwarf agent of wheat Acholeplasma laidlawii var. granulum str. 118 gives rise to pleiotropic responses of acholeplasma: activation of growth process, an increase of common protein in comparison with control, a decrease of hemagglutinating activity which results in a decrease of the adhesion properties of pathogen.

[Elaboration of the in vitro model system to study the interaction of phytopathogenic mollicutes with plant cells].

The model system based on the sugar beet calluses infected by mycoplasms (mollicutes) was elaborated, and changes in the callus cells morphology under the effect of these microorganisms were also studied. The calluses of sugar beet 3K51 cultivated on the Gamborg medium were infected by phytopathogenic mollicute Acholaplasma laidlawii var. granulum str.118. Under the effect of mollicute infection one could observe changes in the cell morphology of sugar beet calluses: the plant cells were transformed from round to lengthened, the intensity of polyploids forming was increased, their grouping and their total destruction were observed. Data of electron microscopy confirm the presence of the mollicute in the sugar beet calluses: acholeplasma cells were localized between and within undifferentiated plant cells.

[Physiological-biochemical and taxonomic studies in mollicutology].

From the beginning of researches in the field of molliculotology and for all the time of existence of special Mycoplasmology Department in the Institute of Microbiology and Virology numerous original results were obtained which are of priority for this science. The collection of strains of various representatives of the class of Mollicutes was formed. Phytopathogenic mollicute strains were first cultived in the elaborated artificial medium and their nature and specific pathogen factors were investigated. Fundamental principles of realization of pathogenic potencies by Mollicutes-agents of plant "yellows" were researched. A number of enzymes of nucleic metabolism and the proteinases, their part as the aggression factors to host organisms was distinguished for the first time, the enzymes localization was studied by cytochemical methods. The composition of carbohydrate part from glycocalix of the mollicutes and microorganisms related to them was studied, and the model of their interaction with the cells of affected organisms was elaborated. The theory and the basis for the practical use of antisignature olygodeoxyribonucleotides as the universal antimicrobial means was formulated. Properties of DNA from mollicutes genome was researched. The system position of these microorganisms and the phylogenetic relations with the representatives of affined taxones was specified. The artificial model system for studying the interaction of phytopathogenic mollicutes with plant cells was created which use helps investigate the particuliarities of the signal and metabolic relations ofmollicutes and cells of host macroorganism. In the course of done researches the changes in the fatty acids composition of the common lipids, in the activity oflectins and enzymes and in the amount of some proteins by the action of mollicute infection were stated.

[Glycopolymers of microorganisms as a source for creation of ecologically-safe antimicrobial media with specific activity]. / Hlikopolimery mikroorhanizmiv iak dzherelo stvorennia ekolohichno bezpechenykh antymikrobnykh zasobiv spetsyfichnoï diï.

Current data concerning structure components of the surface of microorganisms which take direct part in manifestation of their adhesive properties--adhesin, on the one hand, and carbohydrate components of cell receptors, on the other hand, are presented and analyzed in the paper. The prevention or decrease of adhesion by means of competitive inhibition by substances, which enclose structure, analogues of adhesins or their receptors, make up promising trend in the system of treatment and prophylaxis of infectious diseases. The use of synthetic substance of directed action will allow ceasing the development of infectious process without additional chemical loading of the organism-host and pollution of the environment, since as to their structure they resemble biomolecules typical of this organism. It is asummed that the data on the microorganism surface components can serve a basis for conducting fundamental investigations concerning relations between the structure and function of natural biopolymers, specification of phylogenetic relations between certain groups of microorganisms as well as for defining the mechanisms of the action and further perfection of probiotics.

[Discovery of the relationship between carbohydrate determinants of mollicute glycocalyx and affinity of microorganisms, and conditions for their existence]. / Vyiavlennia zv'iazku mizh vuglevodnymy determinantamy glikokaliksu molikutiv i sporidnenykh z nymy mikroorganizmiv ta umovamy ïkh isnuvannia.

As a result of comparative analysis of interaction intensity of plant lectins, specific to certain monosaccharides with surface glycopolymers of philogenetically close groups of microorganisms--mollicutes and representatives of group Bacillus--Lactobacillus--Streptococcus the authors have established common features: availability of alpha- and beta-D-galactose, sialic acids, L-fucose, N-acetylglucosamine, alpha-N-acetyl-D-galactosamine and alpha-D-mannose in their glycocalyx, that is a phenotypical manifestation of genetic relations between these microorganisms. The dependence between the availability of sialic acids and L-fucose in the composition of terminal structures of microorganisms glycocalyx and their capacity to occupy certain ecological niches have been found out.

[The monosaccharide composition of the glycocalyx in Mollicutes and phylogenetically related bacteria in the genus Bacillus]. / Monosakharydnyi sklad hlikokaliksu deiakykh molikutiv i filohenetychno blyz'kykh ïm bakterii rodu Bacillus.

A number of monosaccharides composing the glycocalix carbohydrates of some Mollicutes (Mycoplasma pneumoniae FH, M. hominis PG21, M. fermentans PG18, Acholeplasma laidlawii PG8, A. laidlawii var. granulum st. 118) and phylogenetically related bacteria from the genus Bacillus (Bacillus cereus 89, B. cereus DM423, B. subtilis 1/2, B. licheniformis 31, B. licheniformis 49) were detected using plant lectins of definite carbohydrate specificity labelled by colloid gold. The structure of extracellular glycopolymers of microorganisms was discussed concerning their role in the adhesion process and their specializations in colonization of the appropriate organs and tissues of macroorganisms as well as the action of probiotics on the basis of bacteria from the genus Bacillus. It was marked, that the mollicute cells typically interacted with all vegetable lectins tested while bacilli studied bind certain lectins only. The surface carbohydrates prevailing in monosaccharides from tested one for superficial structures of Bacillus pathogenic strains were: sialic acid, beta-D-galactose and alpha-N-acetyl-D-galactosamine, while L-fucose and alpha-D-glucose were observed in insignificant quantity or were absent. The nonpathogenic Bacillus strains being a basis of biopreparations were distinguished by the insignificant amount of sialic acid and other monosaccharides in the composition of carbohydrates of their glycocalix. Thus, as a result of research it was confirmed, that the superficial glycopolymers of Mollicutes and some related bacilli were alike as to presence of some monosaccharides that depended on taxonomic position of microorganisms and their biological activity.

[The role of the carbohydrate composition of the glycocalyx in some species of lactobacilli in the manifestation of their adhesive properties]. / Rol' vuhlevodnoho skladu hlikokaliksu deiakykh vydiv molochnokyslykh bakterii u proiavi ïkh adhezyvnykh vlastyvostei.

Availability of certain monosaccharides in the composition of glycocalyx of lactic acid bacteria (Lactobacillus plantarum--strains 337D and 11/16; Streptococcus thermophilus--strains S1 (nonmucous race) and S5 (mucous race), Enterococcus faecium (K-50) has been investigated with the help of plant lectins with certain carbohydrate specificity labelled by colloid gold. All the microorganisms under investigation were characterized by the presence of N-acetyl-D-galactosamine and N-acetyl-D-glucosamine in rather insignificant amounts. Glycocalyx of lactic acid bacteria was also characterized by availability of essential amount of L-fructose and low amount of sialic acid (except for S. thermophilus S5 (mucous race). Presence of alpha-N-acetyl-D-galactosamine, alpha-D, beta-D-galactose, alpha-D-glucose, alpha-D-mannose in the composition of the lactic acid bacteria glycocalyx composition evidences for the additional role of these monosaccharides in the process of the microorganism adhesion on the human and animal intestine mucosa. It has been confirmed that availability of certain monosaccharides in the composition of surface glycopolymers of lactic acid bacteria was connected with adhesive properties of cells and their existence conditions.

Antisignature oligonucleotides and their analogs as inhibitors of mollicutes-cofactors of HIV.

Inhibition of mollicutes by synthetic oligonucleotides and their analogs complementary to specific "signature" regions of 16S rRNA and corresponding sequences of ribosomal operon DNA was studied. It was shown that antisignature oligonucleotides inhibited transcription in vitro for above 79% interacting specifically with ribosomal operon and non-specific with DNA-dependent RNA-polymerase. The inhibition efficiency depended on oligonucleotide sequence and type of modification. Translation in vitro was suppressed most efficiently (up to 60%) by oligonucleotides complementary to 3'-end region of 16S rRNA, also depending on their modification. Translation in vivo was inhibited most efficiently (up to 73%) by thiophosphate analogs of oligonucleotides complementary to sequences 499-507 and 523-532 of 16S rRNA responsible for binding of ribosomal "core" protein S4 starting the assembly of 30S ribosome subunit. With the simultaneous use of the last two oligonucleotides, the growth of mollicutes in SM IMV-72 medium rich in exogenous sources of nucleosides was suppressed for over 90%. It is supposed that under conditions where mollicutes have no free access to starting materials for their own synthesis of nucleic acid these nucleotides could suppress microorganisms completely. Antisignature oligonucleotides are considered as superspecific agents not leading to the development of resistance of mollicutes and believed to be the main future remedy against diseased caused by microorganisms lacking the system of nucleoside synthesis.