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2017
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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017, С. 5-13, Bibliography 22, English
https://doi.org/10.15407/biotech10.04.005

Abstract

APPLICATION OF ARTIFICIAL NEURAL NETWORKS METHOD IN BIOTECHNOLOGY O. M. Klyuchko

The aim of the work was to analyze the method of artificial neural networks and to examine its implementation in biotechnology. Nearly 300 publications are reviewed because this method is very widely used. The artificial neural networks are described and analyzed, and the examples of their application in biology and medicine are given. Solutions of complex problems, which required combining this method with other modern mathematical methods, are examined. Recommendations are presented for the application of this method in biotechnology.

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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017
С. 14-24, References, 18, English.
https://doi.org/10.15407/biotech10.04.014

Abstract

ADJUVANT PROPERTIES OF NANOPARTICLES IMMOBILIZED RECOMBINANT DIPHTHERIA TOXOID FRAGMENT T. O. Chudina1, 2, A. Yu.  Labyntsev1, D. V.  Kolybo1, S V. Komisarenko

The aim of the research was to compare the characteristics of nanoparticles with different chemical structure and size (colloidal gold Gold 1 and Gold 2, calcium phosphate CaP and poly(lactideco-glykolid) PLGA 1 and 2) to find the most efficient carriers of antigen — recombinant diphtheria toxoid for per os immunization. According to the MTT test, all studied particles show no significant cytotoxic impact on the studied cells in vitro, with the exception of CaP nanoparticles, which in high concentrations have cytotoxic effect on the U937 cells, and Gold nanoparticles 1 and 2, that are able to inhibit growth of the L929 cells. The most effective phagocytosis by macrophage-like cells J774 is observed for PLGA nanoparticles 1 and 2 with the immobilized antigen, while Gold nanoparticles 1 and 2 with antigen can interact with the surface of these cells without being phagocytated by them. In BALB/c mice immunized per os with antigen immobilized on PLGA 1 and 2 as well as Gold 2 carriers, the concentration of specific IgA antibodies in blood significantly increases after the second immunization, compared with controls. In the group of mice treated with PLGA 2 conjugated antigen, the concentration of specific IgG in blood after the third immunization also increases. These results show the promise of nanoparticles PLGA 1 and 2 as adjuvant for immunization per os.

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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017; С. 25-33, Bibliography 17, English
https://doi.org/10.15407/biotech10.04.025

Abstract

INTENSIFICATION OF MICROBIAL EXOPOLYSACCHARIDE ETHAPOLAN BIOSYNTHESIS ON MIXTURE OF MOLASSES AND SUNFLOWER OIL T. P. Pirog A. A. Voronenko, M. O. Ivakhniuk

The purpose of the research was to establish Acinetobacter sp. IMB B-7005 cultivation conditions, which provide the maximal synthesis of microbial exopolysaccharide ethapolan on a mixture of molasses and sunflower oil, and to explore the possibility of replacing refined oil in a mixture with molasses for waste one. On the basis of theoretical calculations of energy consumption for the synthesis of ethapolan and biomass, it was determined that the optimal molar ratio of the concentrations of energy-deficient (sucrose) and energy-excessive (sunflower oil) substrates in the mixture was 1.0:0.9. Experiments have shown that the highest values of exopolysaccharide synthesis were observed at a molar ratio of monosubstrates in mixture 1.0:1.1, which is as close as possible to the theoretically calculated one. It was shown that increasing concentration of molasses and refined oil in mixture from 1.0 to 1.5% was accompanied by increase in amount of synthesized exopolysaccharide and its synthesizing capacity by 1.2 and 1.3 times, respectively. The possibility of replacing refined oil in a mixture with molasses for various types of waste (after frying potatoes, meat, vegetables and mixed) was established. The maximum parameters of exopolysaccharide synthesis (concentration 14 g/l, synthesizing capacity 3.5 g exopolysaccharide/g biomass) were observed when using mixed waste oil for both inoculum obtaining and EPS biosynthesis. The obtained results testify to the possibility of development of universal technology for obtaining microbal exopolysaccharide ethapolan on a mixture of waste (molasses and waste oil) independent of the type and provider of waste oil.

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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017, С. 34-43, Bibliography 44, English
https://doi.org/10.15407/biotech10.04.034:

Abstract

IRE1 KNOCKDOWN MODIFIES THE EFFECT OF GLUTAMINE DEPRIVATION ON THE EXPRESSION OF A SUBSET OF PROTEASES IN U87 GLIOMA CELLS

O. V. Halkin,    O. O. Riabovol, D. O. Minchenko, A. Y. Kuznetsova,    Ratushna, O. H. Minchenko

The aim of this research was to study the effect of glutamine deprivation on the expression of genes encoding for HTRA1/PRSS11, LONP1/PRSS15, and some cathepsins in U87 glioma cells in relation to inhibition of IRE1 (inositol requiring enzyme-1). It was shown that in control glioma cells (transfected by empty vector) glutamine deprivation up-regulated the expression of LONP1, CTSD, CTSF, CTSO, and CTSS genes, down-regulated HTRA1, CTSC, and CTSK gene expressions, and did not significantly change the expression of CTSA, CTSB, and CTSL genes. Inhibition of ІRE1 signaling enzyme function in U87 glioma cells modified the effect of glutamine deprivation on the expression of HTRA1, LONP1, CTSD, CTSL, CTSO, and CTSS genes: removed the effect of glutamine deprivation on HTRA1 and CTSO genes, introduces on CTSL gene, reduced — on CTSD gene, and enhanced — on LONP1 and CTSS genes. Therefore, glutamine deprivation affect the expression level of most studied genes in relation to the functional activity of IRE1 enzyme, a central mediator of endoplasmic reticulum stress, which responsible for control of cell proliferation and tumor growth.

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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017; С. 44-52, Bibliography. 22, English.
https://doi.org/10.15407/biotech10.04.044

Abstract

VARIANCE ANALYSIS FOR OPTIMIZATION OF THE GERMANIUM BIOLEACHING PROCESS FROM COAL BENEFICIATION DUMPS

I. A. Blaydа, N. Yu. Vasylieva, T. V. Vasylieva, L. I. Sliusarenko

The aim of the work was to optimize the process of germanium bioleaching from the dumps after coal beneficiation, namely, to determine the optimal composition of the new nutrient medium for acidophilic chemolithotrophic bacteria ensuring the maximum recovery of valuable metal in minimum time. We optimized the method of mathematical planning adapted to the plan in Greek-Latin squares. The calculations in this approach are based on the analysis of variance. The formal design of experiments has been carried out with four operating factors at four levels. The calculations were performed in Excel. The significance of the factor levels were analyzed using the Duncan’s multiple range test, the uniformity of the variances was examined the Cochran test, and the significance of the factors was tested by the Fisher criterion for each day of the experiment. The obtained results were interpreted mathematically and biologically. The following combination of factors and their levels was recommended as optimum nutrient medium, g/dm3: KH2PO4 — 1.0; (NH4)2SO4 — 2.0; KCl — 0.1; MgSO4 — 0.5; NH4Cl — 0.5; Na2S2O3 — 5.0. The proposed composition allows the more than 90% quick extraction of germanium into the solution (in four days), which was previously impossible.

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Ж-л "Biotechnologia Acta" Т. 10, № 4, 2017; С. 53-58, Bibliography 17, English
https://doi.org/10.15407/biotech10.04.0593 :

Abstract

BIOMARKERS OF SUBCLINICAL MASTITIS IN THE MAMMARY GLAND OF COWS V. R. Mazurenko, O. V. Manchulyak

The aim of the study was to create an algorithm for controlling subclinical forms of mastitis of cows on the basis of determining the activity of lactate dehydrogenase and the number of somatic cells in milk. Milk samples were taken from conditionally positive cows according to the results of the California test; the activity of lactate dehydrogenase was determined and compared with the content of somatic cells in milk.

According to the results of the analyzes, 2 out of 20 milk samples had low values of lactate dehydrogenase activity, an increased number of somatic cells (more than 250 000 in 1 ml) and negative results of bacteriological examination, which may indicate on the absence of intra-infection and a physiological increase in the number of secreted somatic cells. With increased lactate dehydrogenase activity and a somatic cell level of no more than 250 000 in 1 ml, Streptococcus agalactiae or Staphylococcus aureus bacteria were isolated, indicating on a mono-infection. At the level of somatic cells from 250 000 to 500 000 in 1 ml (4 of 20 milk samples) bacteria Streptococcus agalactiae and Staphylococcus aureus were isolated, indicative on of mix infections.

Thus, the determination of lactate dehydrogenase activity makes it possible to more accurately determine the presence of inflammatory processes in the udder, since the number of somatic cells can also increase with physiological changes (e. g., stress, etc.). The results obtained can be used to determine the subclinical forms of mastitis in the infected herd. Recommendations developed on the basis of this study were implemented in practice in the economy of the Chernihiv region.

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"Biotechnologia Acta" V. 10, No 4, 2017; С. 59-67, Bibliography 18, English.
https://doi.org/10.15407/biotech10.04.059

Abstract

SYNTHESIS OF AROMA COMPOUNDS BY Pleurotus ostreatus (Jacq.:Fr.) Kumm. CULTURED ON VARIOUS SUBSTRATES E. N. Vlasenko, J. V. Stepnevskaya, O. V. Kuznetsova
The aim of the study was to determine the intensity of synthesis of volatile aroma compounds by Pleurotus ostreatus (oyster mushroom) on sunflower husks and barley straw using sensory profile analysis and UV spectroscopy. The main cultural and morphological characteristics of the mycelial growth and development of fruiting bodies are determined: the period of mycelial development on the substrate, the time of primordial formation, the number of mushroom bunches per unit volume of substrate, the morphology of carpophores. Characteristic attributes of the aroma of dried fruiting bodies (mushroom, woody, sweet, herbaceous, fish, meat, floral, earthy, acidic, putrescent) are established and their aroma profiles are built. Sensory profile analysis of flavor of dried samples showed that the mushroom flavor of fungi cultivated on the sunflower husk is more pronounced than of those grown on barley straw. The light absorption maxima are recorded in the ranges 204–210 and 250–290 nm according to UV absorption spectra. Optimal conditions for extracting aromatics from dried fungi samples are the extraction time of 20–35 min at the boiling point of the solvent. Analysis of the UV spectra of fungal alcohol and hexane extracts showed that the intensity of the synthesis of volatile compounds is higher for strains cultivated on sunflower husks than for samples obtained on barley straw.

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MODERN BIOTECHNOLOGICAL APPROACHES TO LIFESPAN EXTENSION OF ANIMALS AND HUMANS E. L. Levitsky

Biotechnologia Acta V. 10, No 2, 2017, Р. 7-21, Bibliography 49, English
https://doi.org/10.15407/biotech10.02.007

Abstract:
The purpose of the research was to analyze current data concerning the problem of extending the life of multicellular animals and humans. The modern views about the processes of aging and prolongation of life are presented. The analysis focused on the genetic mechanisms of aging and mainly biotechnological approaches (genetic engineering, gene therapy, the use of stem cells, and the reprogramming of the genome) to prolong the life of multicellular organisms. For comparison, some traditional methods of prolonging life are described (drug therapy, exercise training, calorically restricted nutrition). This analysis allows   to postulate the perspectives and advantages of using biotechnological methods for prolonging life in comparison with traditional ones.

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INDUSTRIAL WASTE BIOCONVERSION INTO SURFACTANTS  BY   Rhodococcus erythropolis ІMV Ас-5017, Acinetobacter calcoaceticus ІMV В-7241 and  Nocardia vaccinii ІMV В-7405 T. P. Pirog, M. O. Shulyakova, L. V. Nikituk, S. I. Antonuk, I. V. Elperin

Biotechnologia Acta V. 10, No 2, 2017, Р. 22-33, Bibliography 35, English
https://doi.org/10.15407/biotech10.02.022

Abstract:
The aim of the work is to realize an alternative processing of toxic industrial waste into surfactants by strains Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241 and Nocardia vaccinii IMV B-7405 for remediation of environment.The studied strains were grown in liquid media containing such sources of carbon as waste (fried) sunflower oil, technical glycerol (by-product of biodiesel production), and aromatic compounds. The synthesis of surfactants was evaluated by emulsification index, conditional concentration of surfactants and concentration of extracellular surfactants, which was determined gravimetrically after their extraction from supernatant by the mixture of methanol and chloroform. The concentration of oil in water and soil was analyzed by gravimetric method after extraction with hexane.
It was shown that with increasing concentration of the inoculum up to 10−15% and two times increase of nitrogen source content in medium containing 7−8% (v/v) of crude glycerol, concentration of surfactants synthesized by R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B 7241 and N. vaccinii IMV B-7405 was 3.4; 5.0 and 5.3 g/l, respectively, that is 1.6−1.7 times higher as compared with values on basal medium with the same content of substrate. The maximum concentration (3.9−4.3 g/l) of surfactants synthesized by A. calcoaceticus IMV B-7241 on fried sunflower oil (4%) was achieved by using the inoculum grown on refined oil. The ability of R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B-7241 and N. vaccinii IMV B-7405 to decompose aromatic compounds (phenol, naphthalene, toluene, hexachlorobenzene, benzoic and N-phenylanthranilic acid) with simultaneous synthesis of extracellular metabolites with surface-active and emulsifying properties was established. In the presence of surfactants in the form of culture liquid (5−10%), the degree of degradation of complex oil with heavy metal (Cu2+, Cd2+, Pb2+, 0.01−0.5 mmol) pollution in water (3−6 g/l ) and soil (20 g/kg) after 20 days was 82−92%.
Bioconversion of industrial waste into surfactants for environmental technology allows recycling toxic waste, reducing costs of microbial surfactants and provides double effect of environmental purification, which is achieved in the production and use of microbial surfactants.

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ANTIVIRAL ACTIVITY OF LIPOPOLYSACCHARIDES OF Pseudomonas chlororaphis subsp. aureofaciens L .D. Varbanets1, S. L. Rybalko2, D. B. Starosyla

Biotechnologia Acta, V. 10, No 2, 2017, Р. 34-39, Bibliography 9, English
https://doi.org/10.15407/biotech10.02.034

Abstract:
The aim of the study was to investigate the ability of lipopolysaccharides  of two strains of Pseudomonas chlororaphis subsp. aureofaciens to inhibit in vitro the reproduction of human viruses: influenza  A/FM/1/47 (H1N1), herpes simplex type 2 and bovine diarrhea, which is used as a model of hepatitis C virus, as well as to suppress hepatitis C virus production in model system of cells transfected with cDNA of this virus. It has been established that for both lipopolysaccharides in three types of cultures (MDCK, Vero and MDBK) the toxicity is not manifested even in a concentration of 100.0 μg/ml, and decreasing in infectious virus titer more than by 2.0 lg TCD50 (ED99) was already achieved at concentrations of 1.55 mg/ml. Selectivity indexes determination of lipopolysaccharides preparations against the influenza A/FM/1/47 (H1N1) virus, herpes simplex virus type 2 and bovine diarrhea virus shows that lipopolysaccharides of P. chlororaphis subsp. aureofaciens UCM B-306 and UCM B-111 are effective inhibitors of investigated viruses reproduction: selectivity index is at least 64. In the model of Jurkat cells transfected with human hepatitis C virus cDNA, viral RNA loading was determined in cells treated with lipopolysaccharides of P. chlororaphis subsp. aureofaciens. The results of the studies indicate that when lipopolysaccharides of both strains are administered, the production of the hepatitis C virus is completely inhibited.

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WATER BINDING IN COMPOSITE SYSTEMS BASED ON MILLED MEDICINAL PLANTS AND NANOSILICA V. V. Turov, T. V. Krupska, А. P. Golovan, L. S. Аndriyko, М. Т. Кartel

Biotechnologia Acta V. 10, No 2, 2017, Р. 40-56, Bibliography 22, English
https://doi.org/10.15407/biotech10.02.040

Abstract:
The aim of the study was to identify the influence of hydrodensified nanosilica particles on the binding of water by milled plant raw materials in neutral and acidic media. Flowers of Hibiscus sabdariffa and Calendula officinalis were used as the model materials. According to the microphotographs and low temperature 1H NMR spectroscopy data, the silica film forms on the surface of the milled plant particles, and it can significantly influence their hydration. According to the suggested scheme, some of the water from the inner cavities of plant raw materials moves (as evidenced by the decreasing radius of water-filled pores) to the zone of contact of the composite components (the radius of clusters of adsorbed water increases). In studies of desorption of active substances from milled medicinal herbs and their composites by the initial and hydrodensified nanosilica, it has been shown that the formation of a composite significantly reduces the rate of desorption. Minimal desorption is observed in composites containing hydrodensified nanosilica. The studied composite systems are promising for biomedical researches.

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