[Antineoplastic properties of extracellular fructoso-1,6-bisphosphatase of Bacillus subtilis 668 and preparation isolated from cultural liquids of Bacillus subtilis 7025].

It was shown for the first time that extracellular FBPase of B. subtilis 668 like the preparation obtained from culture liquid of B. subtilis B 7025 displays citotoxicity activity in respect of tumor cells of sarcoma 37 in vitro. It is shown that the preparations remove TA antigens from the surface of the tumor cell. It is supposed that the mechanisms of citotoxic effect of extracellular FBFase of B. subtilis 668 and preparation from the culture liquid of B. subtilis B 7025 in vitro on cells of sarcoma 37 is probably realized through the apoptosis.

[Comparative investigation of fructosobisphosphatases of Bacillus subtilis and pale-green dwarfism pathogens of cereals in the reaction of double diffusion in gel according to Ouchterlony].

Serological properties of fructosobisphosphatases (FBPases) of Bacillus subtilis 668 and PGD agent of cereals--the mollicute Acholplasma laidlawii var. granulum st. 118 (Alg 118) were studied in a comparative aspect with the help of the reaction of double diffusion in gel according to Ouchterlony. It was established for each of microorganisms that their extracellular and intracellular enzymes are similar in serologic respect, and each of them is composed of two antigens, one of them being identical in the both microorganisms, while the other displays only partial identity, since it reacts with antibodies in heterological systems with formation of a precipitation line looking as a "spur". That indicates to the fact that antisera to those enzymes contain antibodies both to general determinants of antigens which are compared (FBPases here), and to the determinant absent in one of them. Basing on the investigation results it is concluded that FBPase of B. subtilis is rather similar than identical, in serological aspect, to the enzyme Alg 118 of the same name.

[Effect of silver nitrate, adenosine-5'-monophosphate and phosphoenolpyruvate on activity of extracellular fructose bisphosphatase of Acholeplasma laidlawii var. granulum strain 118].

The reactions of glycolysis or gluconeogenesis proceed in good coordination in the cells of microorganisms, and each stage of these processes is distinctly regulated. Under such conditions fructose-bisphosphatase (FBPase) activity (the enzyme level being constant in the cells of microorganisms) is inhibited by adenosine-5'-monophosphate (AMP) and is activated by phosphoenolpyruvate (PEP) depending on the kind of the source of carbon (glycolytic or glyconeogenic) used for microorganism growth. It is evident that the corresponding regulation of FBPase should be absent in the extracellular environment where one cannot observe a distinct coordination of functioning of the enzyme systems. The investigation results prove that both AMP and PEP, under their individual testing in concentrations up to 20 microM did not practically affect activity of extracellular FBPase, and at higher concentrations they sharply decreased its activity (200 microM AMP by 70%, and PEP - by 75%). Under joint use of PEP and AMP (in concentration 200 microM and 500 microM) one could observe mutual neutralization of the effect of these substances on FBPase; as a result, its activity decreased only by 15% under AMP concentration of 500 microM, and by 25% at AMP concentration of 200 microM, that is in complete agreement with the data of individual testing of the above substances. PEP in high concentrations has displayed itself as a more active repressor of FBPase activity than AMP. AgNO3 in concentrations to 20 microM has manifested itself as a moderate stimulator of FBPase activity and even in the concentration of 200 microM it decreased the enzyme activity by 50% only. The data obtained are rather different than those described in literature for cellular FBPases of microorganisms. It is known that AMP is a powerful inhibitor of its FBPases activity (Ki = 5 microM) while PEP activates it (Ka = 20 microM).

[Effect of separate effectors on activity of extracellular fructosobisphosphatase of Acholeplasma laidlawii var. Granulum, strain 118].

The influence of 16 substances-effectors on the extracellular mollicute fructosobisphosphatase (FBPhase) was studied for the first time. These effectors are used, as a rule, when studying properties of this enzyme biopreparations newly isolated from the cells of animals, plants and cells of microorganisms. It was established that optimum pH for FBPhase of mollicutes is whithin 7.3-7.5 and on the basis of this index it is attributed to the group of the "neutral" of these enzymes. Cations of K, Mn, Mg, NH4, Tl and phosphoenolpyruvate (PEP) increase its activity. Cations of Li and adenosin 5'-monophosphate (AMP) proved to be the inhibitors of mollicute FBPhase activity. Chelators (EDTA, citrate, imidasol pyruvate, L-histidine) activated it inconsiderably (by 10-20%). Cations of Zn, in contrast to those of other tested metals, in low concentrations (0.1 - 0.2 microM) inhibited FBPhase activity, but when increasing their concentration (6 microM and above) activated the enzyme even better than it was observed for Mn and Mg cations, the necessary components of the reacting mixtures. Thus, when determining the components of the reacting mixtures with the purpose to study the properties of mollicute FBPhase and to regulate its activity in the in vitro systems under pH values optimal for the enzyme, the monovalent (NH4, K, Tl) and bivalent (Mg, Mn, Zn) cations may be introduced in their compositions as activators, AMP and Li cations should be used as inhibitors. Other substances which were studied when making their work proved to be inessential effectors is respect of mollicute FBPhase.

[Classification of prokaryotic species: the problem, demands, and methods of solving].

In accordance with the current approaches to the description of species of procaryotic organisms, they should be based on the comprehensive study of morphological, cultural, physiological, biochemical, serological, structure, genetic, molecular-biological, ecological and pathogenic peculiarities of the strains which are described as new species with finding unique and rather stable signs in them for to differ one group of strains from the other one, another set of unique and stable signs being determined for the latter. Thus, a species is a collection of original strains with high level of mutual similarity of the properties, if compare them with another group of relative strains. The strains which have changed key characteristics cannot be identified as the components of the already described species as well as a species cannot be described as based on the investigation of only one strain. Phenotypical signs of the strains composing a species would be identical, and the homology of their genome DNA would not be less than 70%, difference of melting temperatures of these molecules would not exceed 5 degrees C, and identity of sequences of their 16S rRNA is about 96%. The paper deals in detail with all the above propositions with analysis of their state in the world science with classification of procaryotes and specifying methodical and other requirements to the description of these species.

[Molecular weight and subunit composition of extracellular fructosobisphosphatase of Acholeplasma laidlawii var. granulum strain 118].

Molecular weight of extracellular fructosobisphosphatase of Acholeplasma laidlawii var. granulum strain 118--an agent of pale-green dwarf of cereals has been determined. This enzyme is the basic factor of pathogenicity of this organism, and, maybe, of all phytoplasmas, owing to realization of the enzyme noncontrolled function in the plant organism, its habit acquires the disease symptoms characteristic of "yellows". It has been established that in the native condition the extracellular fructosobisphosphatase of Acholeplasma laidlawii var. granulum st. 118 has molecular weight 230 000 +/- 5 000 Da, and in denaturating ones--56 600 Da, i.e., the enzyme in the native condition consists of four equal subunits. The subunit composition is peculiar to fructosobisphosphatase of other microorganisms already described in literature; fructosobisphosphatase of the mollicute as to its parameters both in native and denaturated conditions occupies the intermediate place among them.

[On the path to creation of genealogical (natural) classification of organisms].

C. Woose and his followers have realized the main dream of systematicians and laid the basement for creation of such classification of organisms which whoud allow for their genealogical affinity. Departing from the achievement of molecular biology and genomics and using their own results of sequences of 16S pRNA and their accurate analysis they have constructed the trees of the most probable phylogeny of microorganisms and divided everything living on the Earth into three primary empires (domains): Bacteria, Archaea and Eukarya. These achievements were accepted ambiguously and even negatively by classical systematicians of Darwin's school. The paper deals with the situation in the camp of systematicians, with their objective and subjective approaches to development of this science. Despite the animosity of classical systematicians to the above achievements there is sureness that new achievement will go along the path shown by C. Woose to complete reaching the aim--creation of genealogical (natural) classification of all organisms populating the Earth.

[Classification of mollicutes]. / Klasyfikatsiia molikutiv.

The paper deals, allowing for the newest achievements in the study of mollicute biology, with main principles and characteristics which would to be used in classification of these microorganisms. The present classification of mollicutes is based on the study of classical features of cells (morphology and ultrastructure of cells, morphology of colonies, requirements to nutrition components, biochemical and serological properties) and molecular biological properties (composition of cell proteins, properties of nucleic acids, and especially, ribosomal operons and ribosomal RNA), living conditions, ecological niches, etc. The author hopes that the work will help biologists, physicians, veterinaries, phytopathologists and other persons interested in the study of mollicutes to orient themselves in determining their systematic position.

[Method for obtaining the basic factor of pathogenicity of Phytoplasma--extracellular fructose-1,6-bisphosphatase (on the model of the agent of pale-green dwarfness of cereals in culture)]. / Spoib oderzhannia pozaklitynnoï fruktozo-1,6-bisfosfatazy--osnovnoho faktora patohennosti fitoplazm (na modeli zbudnyka blido-zelenoï karlykovosti zernobykh kul'tur).

A preparation with fructose-1,6-bisphosphatase (FBPase) activity has been isolated as a result of the five-stage treatment of the culture liquid obtained after growing the agent of pale-green dwarfness of cereals on the medium CM IMB-72. After this enzyme treatment by means of hydrophobic chromatography on the column from Toyopearl HW-60 (the enzyme was obtained from the column with decrease of ammonium sulphate (AS) concentration in the eluating buffer to 0.8 M), the preparation deprivation of AS on the column with Sephadex G-10 and substrate-dependent chromatography on the column with CM-sepharose the extracellular 176-fold purified FBPase. Acholeplasma laidlawii var. granulum strain 118 was obtained--the main pathogenicity factor for the agent of cereals yellow.

[Pathogenicity factors of mollicutes and pathogenesis of some human mycoplasmoses]. / Faktory patohennosti molikutiv i patohenez deiakykh mikoplazmosiv liudyny.

The data of pathogenicity factors of mollicutes and mechanisms of the effect of these microorganisms on the organisms they have infected and, first of all, on people occur in countless literary sources. Such data have been accumulated for above 100 year of mycoplasmology existence as science and remain unachievable for most interested specialists. An attempt has been made to generalize in maximum complete volume everything known about pathogenic potential of human and other mollicutes, their pathogenicity factors and mechanisms of their realization on the dramatic changes occurring in the affected organism under the effect of mycoplasmic infection.