Biotechnologia Acta

9-20

Authors: Zaloilo I. A., Zaloilo O. V. , Buchatskiy L. P.

The paper is a review of the literature concerning the most advanced areas of applying the DNA microarrays method in modern fish farming. Particular attention is given to the efficiency of this approach in diagnostics of various stages of viral and bacterial fish diseases. DNA microarrays using is shown to deadly effect for studying of genetic changes in fish under the influence of temperature factors and aquatic pollutants (including feed excess). Various possibilities of DNA microarrays application of DNA microarrays for interspecies and intraspecies identification of individuals in the wild conditions and fish industry are discribed.

40-52

Authors: O. V. Kyrychenko

А microbial biopreparations, particularly for the leguminous, grains, industrial and other agricultural crops of Ukrainian as well as foreign production are analyzed. All biopreparations on the market of Ukraine may be divided into basic groups — growth-activating preparations, phytoprotective preparations, preparations for the accelerated decomposition of plant wastes, preparations for restoration and preservation of soil fertility (depending on the functional activity of a biological agent) as well as info mono- and complex biopreparations (depending on the number of components within them). Approaches to the creation of complex biopreparations and compositions based on biological agents — microorganisms with agronomically valuable properties, fungi, algae, biologically active metabolites of plants and microorganisms as well as microelements in chelated forms are considered to solve the following key environmental goals: getting organic crop production, restoration and preservation of soil fertility. The results of original research on the complex compositions based on nodule bacteria, rhizobacteria, microorganisms — destructors of xenobiotics, microbial (exopolysaccharides, exometabolites) and plant (phytolectins, extract of blue-green algae) metabolites for growth of legumes and grain cultures are presented. It is established that the complex biopreparations and compositions have more effective and stable action as compared to monoculture. The examples of biopreparations of a new generation which are created using the innovative technologies such as IGET Technology (Induced Gene Expression Triggers), LHO-promoter (lipohitooligosaccharides-promoter) technology, TOP-technology (technology osmoprotector) involving achievements of molecular and cell biology, nanotechnology as well as the modern solid carriers, adhesives, lipkogenes, formularies that provide high titer of microbial cells in biopreparations, close contact the bacterial cells with seeds when inoculated and continued prolonged viability of microorganisms on the seeds are given.

63-79

Authors: Parkhomenko Yu. M.1, Donchenko G. V., Chehovskaya L. I., Stepanenko S. P., Mejenskaya O. A., Gorban E. N.

The aim of the research was to test the ability of the drug "Metovitan" to prevent the redox balance disturbance in the tissues and thiamine diphosphate irreversible oxidation upon exposure of ionizing radiation on the body. The rats were subjected to a single exposure of the X-ray therapeutic instrument RUM-17 to create a dose of 0.5, 1.0 and 5.0 Gray. Preparation "Metovitan" was administered at a dose of 25 mg per 1 kg body weight for 22–24 h before irradiation. Contents of thiamine diphosphate, reduced SH-groups and reactive oxygen species in blood and brain were determined using previously described methods. It has been shown that the active form of the thiamine diphosphate content in the blood was decreased depending on the doses (from 0.5 to 5.0 Gray). At the same time the content of thiamine diphosphate oxidized form was increased. Furthermore the critical changes occurred in metabolic processes redox state parameters, namely, the level of free SH-groups was reduced and the level of reactive oxygen species was increased. Similar changes were observed in the brain tissue. The Metovitan single administration to the animals the day before irradiation, at 25 mg per 1 kg of body weight dose, promotes the protection of the intracellular thiamine diphosphate and redox status in animal tissues (blood, brain tissue) from the negative influence of irradiation (at 0.5; 1.0 Gray doses). These results give reason to recommend the drug for treating of the staff that is involved in the elimination of radioactive contamination. To provide the protection from the higher doses, other scheme of the treatment should be probably recommended.

82-91

Authors: Nidialkova N. А., Varbanets L. D., Seifullina І. I., Shmatkova N. V.

The purpose of the research was to investigate the influence of stanum (IV) and germanium (IV) coordination compounds on peptidases 1 and 2 of Bacillus thuringiensis var. israelensis ІМV В-7465 activity. The study of coordination compounds effect on peptidase activity was carried out by complexes with the enzymes incubation and residual activity to the collagen, elastin and fibrin determination. It was revealed the regularity in the influence of different structure complex compounds on peptidases of B. thuringiensis var. israelensis ІМV В-7465 activity. The stanum (IV) complexes with salicyloylhydrazones of aromatic aldehydes increase collagenase and elastase activities. Substituents replacement in aldehyde fragment of stanum (IV) with isonicotinoylhydrazones of aromatic aldehydes complexes by the less polar ones contributed to increase of elastase activity of both enzymes while an absence of substituents enables to increase peptidase 2 fibrinolytic activity. The complexes of germanіum (IV) with isonicotinoylhydrazone of salicylic aldehyde which contain Zn and Co, increased collagenase activity of the peptidase 1 as well as elastase and fibrinolytic activities of the peptidase 2. In general, all tested complexes may be considered as peptidase effectors of B. thuringiensis var. israelensis ІМV В-7465. A difference of complexes effect on activities of the both enzymes is due to the characteristics of the structure of coordination compounds.

98-107

Authors: N. V. Korotkevych, A. Yu. Labyntsev, D. V. Kolybo, S. V. Komisarenko

The goal of work was obtaining of recombinant soluble form of heparinbinding EGF-like growth factor sHB-EGF in eukaryotic cells based on transposon genetic system. We used methods of recombinant DNA and obtaining of eukaryotic cells for producing of recombinant proteins. We have obtained stable eukaryotic producer of recombinant analogue of soluble form of human heparinbinding EGF-like growth factor sHB-EGF based on HEK293 cells. We used transposon based non-viral expression system for sHBEGF expression. Moreover, studies in mice have shown the possibility of using sHB-EGF expressing genetic constructs for in vivo gene therapy of various pathological conditions.

113-121

Authors: Goltsev A. N., Babenko N. N., Gaevskaya Yu. A., Chelombitko O. V., Dubrava T. G., Bondarovich N. A., Ostankov M. V., Klochkov V. K., Kavok N. S., Malyukin Yu. V., Vinnik Yu. A.

The research of the peculiarities of Ehrlich carcinoma growth in vivo after incubation with nanoparticles based on rare-earth orthovanadates of spherical, spindle-like and rod-like shapes under different concentrations was the aim of this study. By immune fluorescence method there were quantitatively assessed the tumor precursors of various differentiation rate on the presence of phenotype markers CD44, CD24, CD 117 and Sca-1. The inhibition of tumor process after pre-treatment of Ehrlich carcinoma cells with nanoparticles of all the shapes and concentrations has been demonstrated. Nano-spindles of 0.875g/l concentration were in a greater extent capable of tumor growth inhibiting that stipulates a maximal survival of tumor-bearing mice. There has been shown a significant re-distribution in growing tumor of the content of the precursors with the mentioned above phenotype markers after pre-treatment of inoculated Ehrlich carcinoma cells with nanoparticles of all the shapes and concentrations. Predictive value of the coefficient of CD44hi to CD117+ cells’ ratio when assessing the anti-tumor therapy was found.

128-134

Authors: Оliynichuk S. Т., Lysak Т. I., Marynchenko L. V.

The purpose of this work is to study the dependence of ethanol accumulation by-products and secondary products (glycerol and propionic acid) during the fermentation in the case of increasing the wort concentration from 12 to 21% by weight of sugar as an example of commonly used in the alcohol industry the commercial dry yeast company “Danisco” and experimental osmophilic strain Saccharomyces cerevisiae DS-02-E, isolated from a concentrated (80% DM) of rye malt wort which spontaneously fermented. The enzyme preparations “AMYLEX 4T”, “ALPHALASE AFP” and “DIAZYME SSF” were used for the liquefaction and saccharification of starch wort). The finished industrial of both yeast strains were added to the fermentation flasks in an amount of 10% by volume of the primary wort. In the mature brew the unfermented carbohydrates content was determined by colorimetric method with anthrone reagent, alcohol — by glass areometer-alcoholometer, acidity — potentiometrically, the concentration of dry matter — by areometer, glycerol content — by photocolorimetry method. In the brew distillate a volatile impurities content, namely propionic acid, was determined using gas chromatography. Statistical processing of the results of three series of experiments were carried out by calculating the arithmetical mean value of 5 measurements, their standard deviations and errors. To determine the probable differences between the mean values were used Student’s t test. Differences were considered statistically significant at P < 0.05. Reduction for accumulation of glycerol (between 38 till 53%) at higher concentrations of nutrient medium in the case of the yeast Saccharomyces cerevisiae DS-02-E as compared with commercial dry yeast, reduction the formation of unwanted by-product of fermentation — propionic acid (up to 34%), a better ability of the experimental strain to accumulate sugar of wort and to accumulate ethanol (up to 0.1–0.25% vol.) were shown. It was concluded that the involvement of other mechanisms for osmoadaptation not related to HOG (high-osmolarity glycerol) way, or less active glycerol synthesis system in response to osmotic stress. The practical significance of research using a new experimental osmophilic yeast strain consists of increasing the depth of substrate utilization and ethanol yield from the starch of grain raw materials that have a positive impact on the economy and ecology of ethanol (bioethanol) production.