Biotechnologia Acta

5-11

Authors: L. M. Bugyna, O. V. Pallah, T.V. Meleshko, V. V. Bati, N.V. Koval, N.V. Boyko

The purpose of this study was to analyze and compare the existing international methods applied for classifying and identifying foods characteristics in a number of modern computerized databases such as Foodex, INFOODS/FAO, EuroFIR, and present our initial results of Regional and National Ukrainian food composition databases creation. In this study theoretical synthesis and deductive analysis were used, literary review of foreign scientific peer-reviewed sources, characteristics of the software LanguaL, DaRiS were presented. The demand for a language-independent thesaurus (LanguaL) and the needs for a practical, field-based food system (INFOODS) led to the attempts to link these tools and create a minimal set of standards and a consistent approach for the food products identifying and analyzing around the world. The examples of this combined approach were “systems mapping” and the “International Interface Standard for Food Databases”. The exploitation of different tools for compiling of the first Regional (100 local products, project) and National food composition databases (53 products of 6 prioritised traditional foods within BaSeFood projects) were reported.

25-34

Authors: Kirpenko N., Leontieva T.

The current state and perspectives of biotechnological use of microscopic algae were analyzed. The main directions of algobiotechnology, due to the physiological and biochemical features of these organisms, the volume of algae production in the world, the types of microalgae that had already been used or had practical prospects, ways of biomass obtaining and productivity increasing of industrial algae cultivation were given. The state of this problem, expediency of algobiotechnology development and prospects of microalgae cultivation in Ukraine were discussed.

45-51

Authors: S. V. Prylutska, I. I. Grynyuk, T. D. Skaterna, L. B. Drobot, N. S. Slobodyanik, O. P. Matyshevska

The aim of this study was to evaluate the toxicity of noncovalent nanocomplex of C60 fullerene with cisplatin (C60-Cis-Pt) against normal cells. The toxicity of the C60-Cis-Pt nanocomplex compared to the free Cis-Pt was studied by estimating kidney human embryonic (HEK293) cells viability using MTT assay and rat erythrocytes resistance to acid haemolysis. It was shown that free 40 µM Cis-Pt changed the morphology and reduced the viability of HEK293 cells, as well as increased the number of haemolyzed erythrocytes compared to the According to the investigated parameters analysis no cytotoxic effects of C60-Cis-Pt nanocomplex was observed at Cis-Pt equivalent concentration. The prevention of Cis-Pt toxic action against normal cells by its complexation with C60 fullerene opens the prospect of nanostructure usage as an effective cytoprotector and a target carrier in tumor cells..

56-63

Authors: A. M. Taranenko, I. O. Nitovska, L. H. Velykozhon, P. D. Maystrov, M. V. Kuchuk, B. V. Morgun

The research was aimed to develop genetic constructs for Agrobacterium-mediated plant transformation, containing the synthetic cry1Ab gene, and their testing through the transformation of tobacco, followed by a molecular genetic analysis of the obtained plants to confirm the transformation event. Basic methods of DNA cloning, Agrobacterium-mediated transformation of Nicotiana tabacum L. by leaf disc method, selection of transformants in vitro, analysis of the transgene presence in plant DNA, detection of cry1Ab gene expression by PCR with reverse transcription were used. In the course of the study, the vectors pCB182 and pCB241 that contained the synthetic gene cry1Ab were constructed. Agrobacterium-mediated transformation of tobacco was carried out by created vectors and regenerant plants containing transgenes in their DNA were obtained. Expression of cry1Ab transgene in the obtained transformants of tobacco by the RT-PCR method was confirmed. As a result of the Agrobacterium-mediated transformation of plants with pCB182 and pCB241 vectors containing the synthetic cry1Ab lepidopteran resistance gene it is possible to obtain transgenic plants with expression of the transgene.

5-28

Authors: O.M. KLYUCHKO, A.Y. BILETSKY

The objective of this review was to analyze the results of electrophysiological studies of some biologically active chemicals in order to determine relationships between their chemical structures and effects that they produce. It was proposed to use these relationships to improve the logical module of biotechnical expert system that works using a module principle based on object-oriented and regression analysis. The data of electrophysiological studies of glutamatergic receptor antagonists, phenol- and indole-derivatives, purified from the spiders’ venoms of Arthropodae species were analyzed in this work. Some characteristics of receptor blocking by these toxins have been used to demonstrate empirical relationships between the chemical structures of antagonists and their electrophysiological effects. Possibilities to apply such relationships for monitoring of harmful environmental pollutants, phenol- and indole derivatives, as well as for developing new methods of their qualitative and quantitative analysis are discussed.

42-62

Authors: M. Gahramanova, M. Rudyk, L. Skivka

Hepatobiliary system diseases represent an important medical and social problem due to increasing morbidity rates worldwide. Liver and biliary diseases are characterized by complex pathophysiology as well as by multi- and comorbidity. The treatment of such diseases necessitates multitarget drug development. The effectiveness of current drugs in the treatment of hepatobiliary disorders remains low and the incidence of side-effects are profound. This actualizes the search and development of highly effective hepatoprotectors with a low incidence of side effects. Medicinal plants potentially constitute a sourse of such preparations. The review summarizes the data concerning mechanisms of hepatoprotective and immunomodulatory effects of medicinal plants and their phytoconstituents. The prospects for the development and use of herbal remedies in the treatment of hepatobiliary diseases are outlined.

72-81

Authors: M. Kryvtsova, J. Košcova, J. Eftimova, M. J. Spivak

The purpose of the work was to study the antimicrobial, antibiofilm-forming, antioxidant and some biochemical properties of alcoholic extracts of Potentilla erecta L. rhizome. The plants for the study were gathered around the village of Luta, Velyky Berezny rayon, Transcarpathia. From the Potentilla erecta L. rhizome, ethyl and methyl extracts were produced. The subjects for study were their antioxidant activity (by DPPH method), total tannin and flavonoids (by spectrophotometric method), and antimicrobial activity (by diffusion-into-agar method). The clinical isolates were isolated with the use of differentially diagnostic nutrient media. The antibiofilm activity of the extracts were tested in standard 96-well microtitration plates. Ethyl and methyl extracts of Potentilla erecta L. rhizome were shown to reveal high antioxidant activity. Antimicrobial activity of the extracts against Staphylococcus genus bacteria and Candida genus fungi was established. The study proved high capacity of ethanol extract for bacterial biofilm destruction. Thus, the study showed the antimicrobial, antioxidant and antibiofilm-forming activity of tormentil ethyl extract against the isolates from the mouth cavities of patients suffering from parodentium inflammatory diseases, which fact contributes to the application prospects of this extract as an active base for mouth cavity hygiene preparations.

89-100

Authors: L. V. Gorbunov, I. V. Petrov, O. V. Zviahintseva

A mathematical model that simplifies the determination of optimal parameters ensuring the maximum viability of frozen-thawed fruit and berry cuttings was developed. Values of the minimum amount of intracellular water η1min, which minimizes the plasmolysis probability, and η2min, which minimizes the probability of intracellular ice formation, were determined with due account for the bioobject heterogeneity. Free water amounts Δη, forming ice crystals inside the cell during cryopreservation of different of fruit and berry varieties, were calculated. The optimal conditions for cutting dehydration (temperature Ti and incubation time t2, minimum amount of intracellular water ηmin) ensuring the maximum viability after drying and low-temperature adaptation to cryopreservation were selected. The individual features of the viability of frozen-thawed cuttings of different species were quantitatively reflected in the free water index Δη. The maximum viability of frozen-thawed birch and blackcurrant cuttings was achieved, when intracellular water was in the bound, vitrified state Δη = 0. The calculated Δη˃0 for cuttings of different varieties of apple- and pear-trees as well as of raspberry-bushes leads to a decrease in the viability, and it is impossible to obtain viable plum, apricot or grape specimens after low-temperature cryopreservation with no bound water ηс at all.

5-18

Authors: Pirog T. P., Kliuchka I. V., Kliuchka L. V.

This review summarizes the published data and own results concerning synergism of antimicrobial activity of essential oils with antibiotics against bacteria of the family Enterobacteriaceae, genera Staphylococcus, Pseudomonas, Acinetobacter; with synthetic antifungal drug fluconazole, against yeast genus Candida; with surfactants of microbial origin, against bacterial and yeast test cultures. The synergistic effect of the complex of essential oils with antibiotics, enzymes, surfactants, etc. on biofilms was considered as well. Mixing essential oils with other biocides allowed to significantly decrease the minimum inhibitory concentrations of each component. The probability of emerging resistance to antibiotics is also reduced in the pathogenic bacteria and yeasts due to the antimicrobial action of essential oils that caused the dysfunction of cellular membrane of microorganisms. The prospects of implementing complex essential oils with antibiotic nisin in the food industry, and with other antibiotics in veterinary medicine are discussed.

27-33

Authors: Kryvtsova M., Bobryk N., Simon L

The aim of the work was to study the soil agrochemical indices, soil microbiocoenosis, in case of growing of energy cultures and based on the mineralization coefficient, to make a conclusion on the speed of mineralization processes in the soils under study. In conditions of continuous field experiment (2011‒2016), the dynamics of soil microbial associations was studied for willow (Salix triandra x Salix viminalis 'Inger') cultivation with application of experimental fertilizers of different types. In the research fertilizers there were used sulfuric urea, municipal biocompost, municipal sewage sludge compost, rhyolite tuff and willow ash. The soil microbiotic communities analysis was conducted by the method of serial dilutions of soil suspension with the use of differentially diagnostic nutrient media: meat-peptone agar, starch-ammonium agar, Ashby medium, potato agar, Czapek Dox medium, starvation agar, Ploskirev medium. The direction of the microbiological processes in the soils determined. According to the results, it was established that the most promising for the purpose of improving the metabolic activitiof the soil (in the growth of energy willow) is a municipal sewage sludge compost and a municipal biocompost. In case of the use of municipal sewage sludge compost, the number of intestinal bacteria, ammonifiers, micromycetes and actinomycetes was doubled as compared with the control. In case of the use of municipal biocompost, the levels of microscopic fungi and cellulolytic bacteria doubled, and those of intestinal bacteria and pedotrophs tripled as compared with the control. While calculating the mineralization/immobilization index, it was shown that the most significant deviation from the control plot was found in the rhyolite tuff treated soil – a decrease by 6 times, and in case of willow ash by 2.3 times, which proved the inhibition of mineralization of the organic substances in the soil.

42-49

Authors: Rashidova А. M., Babazadeh, S. N. Mammedkhanova V. V., Abiyeva E. Sh.

The aim of the work was to study the dynamics activity of lactate dehydrogenase (LDH; EC 1.1.1.27), aconitase (AH; EC 4.2.1.3), NAD-dependent malate dehydrogenase (MDH; EC 1.1.1.37), succinate dehydrogenase (SDH; EC 1.3.99.1) in homogenates and sub-fractions of brain structures of rat prenatally endured hypoxia at the organogenesis stage (in 11−15 days of development) and their role in the formation of compensatory - adaptive mechanisms in brain in postnatal ontogenesis. It was revealed that increasing of lactate dehydrogenase and malate dehydrogenase activity (P<0.001; P<0.01, correspondently) in the brain structures of the rats prevented metabolic disturbances in the regulation mechanisms of biosynthetic and bioenergetics processes in the brain. It has been shown that prenatal hypoxia upregulates aconitase activity in postnatal development and this process, probably, has a reversible character (P<0.01), the highest indices of succinate dehydrogenase activity were noticed in the hypothalamus and cerebellum of 30-day-old rat as compared to the other structures (P<0.001). Based on the data obtained, it can be concluded that hypoxia at the stage of organogenesis leads to a change in the energy supply process of the brain structures and, possibly, is irreversible. Analysis of changes in the enzymatic system in ontogenesis allows us to identify adaptation mechanisms and to assess the dynamics of changes in enzyme activity when the functional state changes, which make it possible to identify adaptive reserves of enzymes LDH, AH, MDH and SDH in brain exposed to hypoxia.

57-64

Authors: Spiridonov V.H., Ushkalov V.O, Chernyshenko T.M., Kolesnikova I.M.,

The aim of the research was to find out the clot formation by the action of chitin derivates. The biochemical and immunologic investigation methods such as obtaining of fibrinogen, chitosan derivates, electrophoresis in PAAG, Western blot analysis, ELISE, the activated partial thromboplastin time and prothrombin time coagulological tests were used in these studies. The next results were obtained: chitin derivatives in equal measure cause the clot formation in whole blood, blood plasma and fibrinogen solution; the fibrinogen precipitate formed as a result of their action, practically does not contain fibrin; chitosan does not cause the activation of coagulation factors and the absence of newly-formed fibrin confirms it; the addition of calcium chloride to fibrinogen solution in concentration-dependent manner inhibits effect of chitosan. Thus, under the action of chitosan, a fibrinogen precipitate forms due to the destabilization of its molecule by the lack of calcium. Absence of fibrin degradation products excludes the possibility of the fibrinolytic system activation and physiological degradation of the clot. It makes no sense to use haemostatic drugs based on chitosan in clinical practice.

5-23

Authors: O. M. Klyuchko, L. P. Buchatsky, O. V. Melezhyk

The aim of the work was to develop electronic database of biological organisms (fishes, viruses) on the base of the methods that have been used before in mathematics, engineering, physics. The methods of object-oriented system analysis, construction of models of subject spaces (problem space, solution space), ER-diagrams, of constructing databases and others were applied. The examples of prototypes of modern databases with information about fishes developed over the past years in Western European and American countries were presented. An overview of applied methods, concepts and terminology from the areas of object-oriented system analysis, database designing was disclosed. In conclusions the description of constructed database and practical recommendations for the development of databases with the information about domestic biological organisms for electronic information systems are given.

41-49

Authors: V. Sokolik, O. Berchenko, N. Levicheva, S. Shulga

The aim of the study was to determine the effect of miR-101 on the level of β-amyloid peptide and activation of the cytokine system in the brain regions of animals with an experimental model of Alzheimer’s disease. MiR-101 is the key deactivating operator of mRNA function for the amyloid-β protein precursor. Hence, miR-101 is capable to suppress its synthesis and amyloidogenic processing. Aged male rats were injected intrahippocampally with single-dose unilaterally of β-amyloid peptide 40 aggregates (15 nmol). After 10 days, nasal administration of the liposomal form of miR-101 or empty liposomes was started. After 10 days of therapy, the level of toxic endogenous form β-amyloid peptide 42 and the activity of the cytokine system were determined by the indicators of tumor necrosis factor α, interleukin-6, and interleukin-10 in neocortex, hippocampus and olfactory bulbs. It was found that in rats, aggregates of exogenous β-amyloid peptide 40 model the amyloidogenic and pro-inflammatory situation after 20 days in the neocortex and hippocampus (a significant increase in the concentrations of β-amyloid peptide 42 by 36% and cytokines by 16–18% in the neocortex, and β-amyloid peptide 42 by 27%, proinflammatory cytokines tumor necrosis factor α, interleukin-6 by 14% in the hippocampus), but not in olfactory bulbs. The ten-day course of nasal therapy of liposomal miR-101 normalized the level of β-amyloid peptide 42 and cytokines: in neocortex, the concentration of endogenous toxic β-amyloid peptide 42 decreased by 33%, in the hippocampus by 15%, and concentration of pro-inflammatory cytokines fell by 11–20%. Thus, nasal therapy of miR-101 in liposomes caused a significant anti-amyloidogenic effect in rats with the Alzheimer’s disease model, whereas its anti-inflammatory effect was primarily due to a decrease in β-amyloid peptide 42 concentration.

57-66

Authors: M. Kryvokhyzha1, Y. Libantova2, N. Rashydov

The aim of the work was to estimate the impact of the short wavelengths ultraviolet radiation (wavelength is 230 nm) on Arabidopsis thaliana. The stress response on some key flowering determination genes AP1, GI, LFY, FT, CO, and the repair gene RAD51 expression were investigated. The grown plants were applied by red (610–700 nm), violet (400–450 nm), neutral white (mixture wavelengths 380–750 nm), 20 V and high intensive white light (mixture wavelengths 380–750 nm) 40V LED. The experimental group of plants was irradiated by short wavelengths ultraviolet on ontogenesis stage 5.1 by Boyes classification. The leaf length as growth parameter mark also was analyzed. The short wavelengths ultraviolet influence caused differences in photoperiodic pathway genes expression in plants grown under different illumination. Acceleration flowering phases under influence white intensive illumination and delay ones in case of violet and common white illumination were observed comparing with control groups. It was revealed that cryptochrome and phytochrome formation play an important role in plant development and stress resistance. It enables to understand the best way of plant cultivation in stressful condition.

75-81

Authors: О. Hrytsev, J. Shevchenko, N. Vorobiova, L. Skivka

The aim of the study was to develop rapid and sensitive assay for the simultaneous detection of R. cerealis and R. solani. Pure cultures of fungi were grown on a potato dextrose agar for 5 days at 28 °C, and mycelium was harvested and used for DNA extraction. Total DNA was extracted using a commercial test-systems. Molecular identification of phytopathogenic fungi was performed using a multiplex-touchdown PCR with further electrophoretic separation of amplification products in agarose gel. The specific sequence characterized amplified region primers RtubR4/RtubF4 for R. cerealis and ITS1/GMRS–3 for R. solani were tested for their specificity and useability in PCR multiplex capacity. The specificity of the multiplex-touchdown PCR was tested using DNA from wide range of fungal species and non-target DNA from healthy wheat. The used primer pairs provided only specific fragments for R. cerealis and R.solani. No PCR products were obtained during amplification with the negative control or non-target DNA templates from other species. Coupled to this we have optimized the temperature regime for the multiplex PCR protocol. Taken together, our protocol convincingly demonstrated the simultaneous ability to detect Rhizoctonia cerealis and Rhizoctonia solani and can be used for the diagnosis of compound Rhizoctonia root rot.

5-26

Authors: Klyuchko O.M., Aralova A.A., Aralova N.I.

The purpose of the work was to analyze data about the construction of electronic automated work places for organization of more perfect ones for biotechnology. Methods of program and mathematic simulation, imitation modeling were used for these works. The information about some prototypes of electronic automated work places constructed for biology and linked sciences in Ukraine during the last 25–30 years was discussed. The results of some automated work places constructed by the authors were presented. In conclusion the observed experience was summarized and the set of recommendations for its practical implementation were done.

46-55

Authors: M. M. Marchenko, I. V. Dorosh, L. M. Cheban

The aim of the study was to develop the biotechnology approach for obtaining secondary carotenoids of green microalgae Desmodesmus armatus (Chod.) Hegew. under conditions of two-stage cultivation on waste water from recirculating aquaculture system in response to the action of inducers of different origin. By chemical nature, secondary carotenoids are C40-ketocarotinoids - intermediates of enzymatic oxidation of β-carotene to astaxanthin. The study presents the conditions for cultivation of D. armatus on the waste water from the recirculating aquaculture system by a two-stage accumulation process, where conditions for rapid growth of biomass were created at the first stage, and the biosynthesis of the target product was induced by the introduction of carotenoid biosynthesis precursors (C6H12O6, CH3COONa), promoters of free radical oxidation (FeSO4 / H2O2) or osmotic stress (NaCl) into the nutrient medium. It was shown that the first phase of cultivation was characterized by high growth and productive indices: the amount of biomass was up to 13 g/l, the content of total proteins was 37.9 %, lipids – 26 % and total carotenoids – 7.5 % per gram of dry biomass. Among carotenoids, the presence of zeaxanthin, lutein, β-carotene, insignificant amounts of astaxanthin, canthaxanthin, esters of adonixanthin and astaxanthin were detected. The features of the adaptive response of D. armatus to the influence of factors that induce secondary carotenogenesis are established. Among them is retention of the number of cells or doubling of their number during the use of chemical activators. Decrease in the activity of cytochrome oxidase as an indicator of the metabolic activity of the culture. Thus, the possibility of increasing the content of β-carotene and astaxanthin in D. armatus biomass, essential for fish and crustaceans, by introducing promoters of free radical oxidation and osmotic stress NaCl (200 mM) or Fe2+ (200 mM) and H2O2 (10–4 mM) into the waste water from RAS in the second phase of cultivation was established. Metabolic disbalance in D. armatus cells, which were observed under the influence of chemical factors, led to a redistribution of the main nutrients profile. Biosynthesis and accumulation of lipids were activated against the background of intensive carotenogenesis.

63-70

Authors: M. Gahramanova, 2, R. Dovhyi, M. Rudyk, O. Molozhava, V. Svyatetska, L. Skivka

The aim of the work was to explore phytochemical characteristics of water extract from polyherbal composition based on P. oleracea and it’s effect on oxidative metabolism of murine peritoneal macrophages. The qualitative phytochemical analysis was conducted by colorimetric method. Quantitative analysis of phenols was performed in the test with gallic acid as a standard. Murine peritoneal macrophages were isolated without previous sensitization. Leukotoxicity of the water extract from polyherbal composition leukotoxicity was evaluated in MTT test. Reactive oxygen species generation was assayed by the nitroblue tetrazolium reduction method. Phytochemical analysis revealed the presence of water-soluble and insoluble phenols, tannins, saponins, flavonoids, cardiac glycosides and coumarins in the studied plant mixture. The water extract from polyherbal composition used in a range of concentration 1–1000 μg/ml (lyophilisate in distilled H2O) didn’t exhibit any toxic effects on murine peritoneal macrophages. Water extract from polyherbal composition caused statistically significant dose-dependent increase in oxidative metabolism of murine peritoneal suggest modulatory effect of studied water extract from polyherbal composition on innate immunity cells.

79-87

Authors: O. V. Kyrychenko

The aim of the work was to estimate the action efficiency of pre-sowing soybean seed bacterization with complex inoculants based on Bradyrhizobium japonicum 634b and Azotobacter chroococcum Т79 under influence of phytolectins in vegetation conditions. It was shown, that during all vegetation period the soybean plants formed vegetative mass more actively: (in 1.2–1.5 times) above-ground part and in 1.2–1.7 times root system by the the complex seed bacterization as compared to the mono-inoculation. There is a direct dependence of soybean vegetative height on the functional (nitrogen-fixing) ability of the symbioses. Advantages of the application of complex compositions for intensification of beans formations (more early terms of reproductive organs forming, greater amount of beans on plants with their mass, exceeding control in 1.1–1.7 time) are shown. The middle increase of soybean harvest to control made from 13% (binary bacterial composition on basis of rhizobium and azotobacter) to 21% (polycomposition on basis of rhizobium and azotobacter activated by the wheat lectin). The compositions based on rhizobium activated by the soybean lectin provided 18% increased seed harvest. Polycomposition containing nitrogen-fixing bacteria activated by appropriate plants lectins led to the 19% increased harvest. It is shown that the harvest increased with higher values of almost all indexes of its structure. The compositions based on rhizobia and azotobacter activated by wheat lectin as well as the compositions based on rhizobia activated by soybean lectin are the most productive for practical use to increase the soybean yield.