Biotechnologia Acta

9-16

Authors: O. I. Krynina, D. V. Kolibo, S. V. Komisarenko

The comparative analysis of 8 antigens of Mycobacterium tuberculosis (MPT83, MPT63, ESAT-6, CFP10, SodA, leuD, panD and HBHA) in silico to determine T-epitopes that can bind MHC class II molecules coding the most common in Eastern Europe HLA-DRB1 alleles was the goal of the work. The results show that the most immunogenic proteins are SodA, MPT83, leuD and MPT63, which has the largest number of T-epitopes for the alleles of interest. Accordingly, these antigens or their combinations could be promising candidates for development of new vaccines and diagnostics to form anti-TB immunity in Eastern European region and to determine the state of protection of the population.

29-39

Authors: O. V. Ostrynska, O. P. Kukharenko, V. G. Bdzhola, S. M. Yarmoluk

The effect of protein kinase CK2 inhibitors (with IC50 from 0.004 μM to 0.7 μM) from different chemical classes on the activity of СК2α and СК2α′ recombinant proteins has been studied. Biochemical tests shown that isozymes had different sensitivity toward the same compounds. The most isoform-selective inhibitor was 4′-hydroxyflavone derivative (FLC26) with IC50 value 0.020 μM (CK2α) and 0.003 μM (CK2α′). To explain the difference between influences of FLC26 on the activities of two CK2 catalytic subunits their complexes with ATP-binding site of CK2α and CK2α′ were analyzed using molecular modeling techniques. The data obtained by molecular dynamics simulation (10 ns) has not provided a clear explanation of the difference between the inhibitory potency of the compound FLC26 towards the CK2 catalytic subunits. Thus, the reasons underlying the different activities of the same inhibitor on CKα and CKα′ require further investigations. An effective approach for this purpose would be X-ray analysis of complexes «compound FLC26- CKα/CKα′».

46-50

Authors: V. V. Klochko

Biosynthesis of antistaphylococcal antibiotic batumin under periodic conditions of Pseudomonas batumici growth has been studied. Antibiotic synthesis in fermenter occurred across the culture growth and achieved its maximal value after 50–55 hours. The active oxygen utilization by the producing strain was observed during 20–55 hours of fermentation with maximum after 40–45 hours. Antibiotic yield was 175–180 mg/l and depended on intensity of aeration. contrast to «freshly isolated» antibiotic after fermentation the long-term kept batumin has shown two identical by molecular mass peaks according to the chromato-mass spectrometric analysis. Taking into account of batumin molecule structure the conclusion has been made that the most probable isomerization type is keto-enolic tautomerism. At the same time batumin is diastereoisomer of kalimantacin A which has the same chemical structure. The optic rotation angle is [α]d25 = +56.3° for kalimantacin and [α]d25 = –13.5° for batumin. The simultaneous P. batumici growth and antibiotic biosynthesis and the ability of this molecule to optical isomerisation and keto-enolic forms formation allow us to suppose that batumin plays a certain role in metabolism of the producing strain.

57-62

Authors: B. A. Rasulov

The aim of the research was to obtain silver nanoparticles and to study of its antibacterial activity against pathogenic cultures Escherichia coli ATCC11229 and Staphylococcus aureus ATCC6538. Obtained complex was characterized by UV-Vis spectroscopy, X-ray diffraction (XRD) and transmission electron microscopy (TEM). Colloid solutions of the complex absorbed radiation in the visible regions of 420–460 nm, which confirmed the formation of silver nanoparticles. The size of synthesized SNP varied from 6 to 50 nm. There has been confirmed presence in them of atomic silver. Obtained compound possessed antibacterial activity against pathogens, such as Escherichia coli ATCC11229 and Staphylococcus aureus ATCC6538.

63-68

Authors: K. V. Liapina, P. G. Dulnev, A. I. Marinin, T. V. Melnichenko, A. I. Ustinov

The aim of the work was obtaining a stable suspension based on biocompatible substances with application of biogenic metal nanoparticles encapsulated into NaCl salt matrix, as a precursor. Water-soluble complex based on different amine derivatives with antiseptic properties was selected as a liquid for salt dissolution. The solution was subjected to dispersion using ultrasonication at elevated temperature. Dispersion is accompanied by salt shell removal with simultaneous formation of an organic shell on the surfaces of metal nanoparticles that ensure their stabilization. Study of the suspension after soaking at room temperature for 100 days showed that its characteristics remain stable. A method for producing a stable colloidal solution based on nanoparticles of biogenic metal (Cu, Co, fem etc.) was developed. Metal nanopowder encapsulated into salt shell was used as a precursor. It is shown that such colloidal solutions are characterized by narrow size dispersion, as well as stability to temperature impact and time factor.

83-91

Authors: I. S. Chekman, N. A. Gorchakova, H. O. Sirova, O. O. Kazakova, T. I. Nagorna, V. F. Shatornaya

Review is devoted to the amino acids that could be used for nanostructures creation. The investigation of corresponding properties of amino acids is essential for their role definition in creation of nanomedicines. However, amino acid studying as components of nanostructures is insufficient. Study of nanoparticles for medicines creation was initiated by the development of nanotechnology. Amino acids in complexes with the nanoparticles of organic and inorganic nature play an important role for medicines targeting in pathological process. They could reduce toxicity of the nanomaterials used in nanomedicine and are used for creation of biosensors, lab-on-chip and therefore they are a promising material for synthesis of new nanodrugs and diagnostic tools.

102-108

Authors: A. V. Oberemko, A. G. Popandopulo

This review presents a generalized definition of vesicles as bilayer extracellular organelles of all celular forms of life: not only eu-, but also prokaryotic. The structure and composition of extracellular vesicles, history of research, nomenclature, their impact on life processes in health and disease are discussed. Moreover, vesicles may be useful as clinical instruments for biomarkers, and they are promising as biotechnological drug. However, many questions in this area are still unresolved and need to be addressed in the future. The most interesting from the point of view of practical health care represents a direction to study the effect of exosomes and microvesicles in the development and progression of a particular disease, the possibility of adjusting the pathological process by means of extracellular vesicles of a particular type, acting as an active ingredient. Relevant is the further elucidation of the role and importance of exosomes to the surrounding cells, tissues and organs at the molecular level, the prospects for the use of non-cellular vesicles as biomarkers of disease.

27-34

Authors: Nidialkova N. А., Matseliukh О. V., Varbanets L. D.

The cultivation conditions of Bacillus thuringiensis ІМV В-7324 for synthesis of the elastolytic peptidase were studied. By mono- and two-factorial experiments it was optimized the nutrient medium and conditions of growth for the synthesis of B. thuringiensis ІМV В-7324 elastolytic peptidase. It was established that maximal synthesis of the enzyme (5.15 U/mg of the protein) occurs at the exponential phase of growth on 18 h in submerged cultivation. As a result of screening experiments it was shown that all components of the basic medium except gelatin are significant for the enzyme biosynthesis. The elimination of gelatin leads in 9.8-fold increase of the elastolytic activity (50.55 U/mg of the protein). The influence of the nitrogen and carbon sources on the enzyme synthesis was studied. It was established that the optimal sources are the ammonium sulfate and arabinose. Their usage allows us to increase in 17.4 and 4.6 times the elastolytic activity (90 and 24 U/mg of the protein). The optimal concentrations of the ammonium sulfate and arabinose in the medium which allow to increase the elastolytic activity in 24.7 times (127.45 U/mg of the protein) was determined by the bifactorial experiment on three levels. The optimized nutrient medium contains (g/l): arabinose — 13.0; (NH4)2SO4 — 14.0; КН2РО4 — 1.6; (CH3COO)2Zn — 0.25; MgSO4∙7H2O — 0.75. The elastolytic activity of B. thuringiensis ІМV В-7324 was 132.5 U/mg of protein during the growth by submerged cultivation for 18 h in 200 ml of the optimized nutrient medium at initial pH 7.0, on a shaker at 220 rpm at 37 °C.

43-49

Authors: Tashyrev O. B., Suslova O. S., Rokitko P. V., Oleksenko H. O., Bondar K. M.

To determine the studied bacteria resistance quantitative parameters of extreme factors such as toxic metals (Cu2+), organic xenobiotics (p-nitrochlorobenzene) and UV-irradiation were the aim of the research. Six strains of nitrogen-fixing bacteria isolated from clays of two caverns Mushkarova Yama (Podolia, Ukraine) and Kuybyshevskaya (Western Caucasus, Abkhazia) and Azotobacter vinelandii УКМ В-6017 as a reference strain have been tested. For this purpose the maximum permissible concentration of Cu2+ and p-nitrochlorobenzene in the concentration gradient and lethal doses of UV by the survival caverns have been determined. Maximum permissible concentrations for strains were as 10 ppm Cu2+, 70–120 ppm of p-nitrochlorobenzene. The maximum doses of UV-irradiation varied in the range of 55–85 J/m2 (LD99.99). It is shown that three classes of extreme factors resistance parameters of karst caverns strains are similar to the strain of terrestrial soil ecosystems. The most active studied strains reduce the concentration of p-nitrochlorobenzene in the medium in 13 times. The ability of nitrogen-fixing bacteria to degrade p-nitrochlorobenzene could be used in creation new environmental biotechnology for industrial wastewater treatment from nitrochloroaromatic xenobiotics. Isolated strains could be used as destructors for soils bioremediation in agrobiotechnologies and to optimize plants nitrogen nutrition in terrestrial ecosystems.

55-61

Authors: Shulga S. M.

Creation of liposomal forms of curcumin (diferuloilmetane) was investegated in the work. The methods of preparation and determination of the obtained liposomes sizes and percentage of curcumin in liposomes are given. Soybean phospholipids, egg yolk phospholipids and sunflower phospholipids were selected for preparation of “empty” liposomes and liposomes loaded curcumin: C-SPC-L (curcumin encapsulated in SPC liposomes), C-EPC-L (curcumin encapsulated in EPC liposomes) and C-SFPC-L (curcumin encapsulated in liposomes SFPC). Physical properties of different liposomes such as average size (Dl) and encapsulation efficiency (EE) were studed. Using photon correlation spectroscopy, the average size of three types of curcumin-loaded liposomes were found as Dl = 111 ± 3 nm for C-SPC-L, Dl = 98 ± 5 nm for C-EPC-L and Dl = 109 ± 5 nm for C-SFPC-L. Encapsulation efficiency showed the following values: EEC-SPC-L = 83 ± 4%, EEC-EPC-L = 82 ± 3%, and EEC-SFPC-L = 85 ± 4%. We determined that by using the methods of freezing and thawing or sonication it was possible to isolate liposomes sized in the range 100– 120 nm. Upon that the obtained liposomal form of curcumin was stable and freeze-thaw method was optimal for liposomal form and provides a relatively high level (more than 80%) of curcumin inclusion.

71-76

Authors: Tevfik A. Sh., Mitrofanova I. V., Kuzmina T. N.

Regenerants of Canna (Canna × hybrida hort.) cvs. Suevia, Dar Vostoka and Livadia have been obtained. Dependence of viability and regenerative capacity of Canna microshoots on genotype has been found. The optimal concentrations of growth regulators that influence on morphogenesis in vitro have been determined. At the micropropagation stage of different cultivars of Canna it needs the addition of 1.24–1.91 mg/l thidiazuron to culture medium. The active rooting of Canna microshoots cvs. Dar Vostoka and Suevia on the culture medium Murashige & Skoog and Pierik with auxins β-indol-3- acetic acid and α-naphthylacetic acid have been observed. The ability of Canna cv. Suevia microshoots to spontaneous root formation has been shown. The formation of meristemoids in culture in vitro of Canna explants have been confirmed by the results of histological investigations.

85-93

Authors: Naumenko О. V.

Search of biologically active Lactobacillus strains prospective for functional milk food production was the aim of the research. The study involved the lactic acid bacteria isolated from biological material of healthy humen and non- dairy lactic products. Using modern methodological approaches, the strains of lactic acid bacteria such as Lactobacillus casei 302, Lactobacillus acidophilus 35 and Streptococcus thermophilus 21 having high level of biological activity were selected. High biological potential of selected cultures of lactic acid bacteria, which could provide stability for the technological process of production and essential characteristics of bacterial preparations and fermented their products, was set. In vitro the experiments demonstrated that selected strains had valuable production properties, namely the ability to reduce level of cholesterol and lactose during development in milk, were resistant to virulent bacteriophages and aggressive compounds of the gastrointestinal tract, and high adhesive and antagonistic activities as well.

101-107

Authors: Kosakivska I. V., Babenko L. M., Skaternya T. D., Ustinova A. Yu.

The effects of temperature regime on the pigment spectrum and lipoxygenase activity of the frost-resistant winter wheat Triticum aestivum L. cv. Volodarka seedlings were analyzed. After short-term cooling (+4 °C, 2 h) the amount of chlorophyll a in 7-day-old seedlings decreased. The chlorophylls a+b/carotenoids ratio increased under high temperature treatment (+40 °C, 2 h.) from 10.9 to 18.5. In 14-day-old seedlings a low temperature resulted in some rise of chlorophyll a content, the chlorophylls a/b ratio raised from 2.13 to 2.97 while the content of chlorophyll b and carotenoids diminished. Two isoforms of lipoxygenase: LO-1 (рН 7.0) and LO–2 (рН 6.0) were revealed in the seedling leaves, in roots — one 9-LO (pH 6.5). After high temperature and cross stress lipoxygenase activity increased both in the leaves and in the roots. After cooling 9-LO from roots some decrease of its activity was shown. The revealed changes in the pigment spectrum and lipoxygenase activity are regarded as components of adaptive response to changes in temperature regime. These results open the possibility to use the quantitative ratios of photosynthetic pigments and lipoxygenase activity as markers for selection for creating new high technology crop varieties.

114-119

Authors: Bal-Prilipko L. V., Leonova B. I.

The analysis of literature and patents related to the possibilities of biotechnology for optimizing the domestic meat processing plants was the aim of the article. The analysis of the results of the use of biotechnological methods in the meat processing industry is given. The prospects for their implementation are evaluated. The main development strategy of technological meat processing to develop the methods of obtaining high quality and safe meat products is highlighted. Targeted use of special strains of microorganisms in production of functional meat products offers some opportunities. Thus, such action is associated with formation of the following specific dietary components: organic acids, bactericins, enzymes, vitamins and others. They promote to improve the sanitary microbiological, organoleptic, functional and technological parameters of meat products. Using of denitrifying microbial strains could reduce the residual content of sodium nitrite in the finished product, minimizing the possible carcinogenic and mutagenic impact of this compound on a human body, producing functional safe products while maintaining its high organoleptic characteristics.

25-34

Authors: Garda S. A., S. G. Danilenko, G. S. Litvinov

Probiotics based on normal microflora of the birds using perspective strains become increasingly popular for treatment and prophylaxis of dysbacteriosis in poultry. The purpose of the work is the biotechnological data analysis of the composition and functions of the microflora of different birds’ biotopes. One of biotechnological methods for the study of bacterial flora in the birds is a method of in vivo bacteriological control — analysis of group samples of fresh droppings. To study bird bacterial microflora the method based on vital bacteriological control (group sample study of fresh brood) is the most effective. Only 60–70% of microorganisms are identified during the analysis of bowels bird microflora. It is shown that the normal microflora of the birds has a protective function because it is colonized on epithelial intestinal area and competes for power sources, has a wider set of enzymes, and also produces a wide range of exometabolites that determine their antagonistic action on pathogenic and conditionally pathogenic transient microorganisms. To improve modern technologies concerning cultivation of various breeds of birds with high genetic potential it needs full understanding of endogenous microflora role in a bird body. We found that as a source of probiotic strains it is better to use gastrointestinal tract laying hens and/or to make a selection of group tests of their fresh litter. Thus the best probiotic properties are characterized by microorganisms genera Bifidobacterium and Lactobacillus. The results could be used for selection of promising strains to create a acomplex probiotic.

43-48

Authors: Gudzenko E. V.. Varbanets L. D., Kurchenko I. M., Naconechnaya L. T.

The purpose of this work was to investigate α-L-rhamnosidase that hydrolytically cleaves the terminal unreduced α-1,2-, α-1,4- and α-1,6-linked rhamnose residues in both synthetic and natural glycosides, oligo-, and polysaccharides, various glycoconjugates: flavonoid derivatives such es rutin, neohesperidin, hesperidin, naringin, quercitrin, saponins, terpene glycosides. These properties of the enzyme could be used for the needs of food industry, pharmaceutical and chemical industry: to improve the quality of beverages (reduction of bitterness, flavor enhancing wines), for production of food additives, medicine preparations and rhamnose. As a result of screening conducted among 9 strains of micromycetes, ability to synthesize α–Lrhamnosidase was revealed only in Penicillium sp. 2918. Complex enzyme preparation was obtained from culture supernatant of this micromycete by fractionation with ammonium sulfate (90% saturation) and its physico-chemical properties such as pH- and thermooptimum, pH- and thermal stability and substrate specificity were studied as well. It is shown that enzyme has pH optimum is about 6.0 and thermooptimum is about 60 оC. Preparation of Penicillium sp. 2918 with α-L-rhamnosidase reveals α-D-glucosidase, α-D-galactosidase and α D-glucosaminidase activity.

54-60

Authors: Tigunova O. O., Shulga S. M.

The purpose of our work was to study generic and specific affinity of domestic strainsproducers, comparative productivity butanol strains-producers screening, fermentation of sugars being a part of renewable lignocelluloses raw materials and to determine the conditions for the butanol yield increasing. The objects of research were strains Clostridium acetobutylicum ІМВ В-7407 (IFBG C6H), IFBG C4B and IFBG C7P from «Collection microorganism’s stains and plants line for food and agriculture biotechnology» of Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine. It was determined that domestic butanol-producing strains were relatively more productive and might be promising for improvement technology of butanol production.

71-79

Authors: R. G. Vasyliev, A. E. Rodnichenko, D.A. Zubov, I. F. Labunets, S. N. Novikova, G. M. Butenko

A culture method for multipotent neural crest-derived stem cell isolated from the bulge region of the hair follicle of whisker pad of adult mice has been described and their biological properties have been studied. It was shown that the cells possess a fibroblast-like morphology, they are nestin-positive and cytokeratin-negative, and also express the following surface markers: CD44, CD73, CD90 and Sca-1. This cell type shows the functional properties of stem cells in culture: clonogenicity, self-renewal, sphere-forming capacity and the ability to the directed multilineage differentiation. Due to these properties, neural crest-derived multipotent stem cells are promising for application in the regenerative medicine

85-91

Authors: Gorbach O. I., Khranovska N. M., Skachkova O. V., Sydor R. I., Pozur V. K.

The aim of the study was to develop a scheme of combined chemoimmunotherapy and to investigate antitumor and immunomodulatory activity of chemoimmunotherapy regimen using the vaccine based on dendritic cells and low-doses of cisplatin in CBA mice with sarcoma-37. Maximal antitumor and immunomodulatory effects were observed after application of the vaccine based on dendritic cells in combination with doses of cisplatin concentration of 2 mg/kg. Among significant immunomodulatory effects of combination therapy it has to be noted the increased functional activity of natural immunity,in particular, enhancing of cytotoxic activity of natural killer cells and the ability of peritoneal macrophages, neutrophils and spleen macrophages to increase their absorbing activity and to produce the active oxygen forms. The obtained results prove the expediency of combining of chemo- and immunotherapeutic methods for the development of more effective approaches to prevent recurrence and metastasis after primary treatment of cancer patients.

100-106

Authors: Ryazantsev V. V., Babijchuk L. A., Mykhailova O. O., Zubov P. M.

The generation of reactive oxygen species in different populations of nucleated cells during cord blood cryopreservation was studied. Using cytometry with the 2′,7′-dichlorodihydrofluorescein diacetate dye we assessed the integral content of reactive oxygen species in various populations (lymphocytes, monocytes, granulocytes) of cord blood nucleated cells, depending on the method of isolation, treatment with cryoprotectant and freezing. Furthermore the activity of antioxidant enzymes prior to and after cryopreservation while the cells were loaded with exogenous reactive oxygen species, which source was hydrogen peroxide, was assessed. The results obtained showed that the isolation and cryopreservation of cord blood nucleated cells according to proposed method, which included the isolation of cells in polyglucinum and freezing with 5% DMSO did not result in a significant increase in level of intracellular reactive oxygen species. In addition the activity of antioxidant enzymes to exogenous hydrogen peroxide inactivation in cryopreserved cell was maintained at the level close to that in the native cord blood. Among all the populations of cord blood nucleated cells the granulocytes were the least resistant to cryopreservation effects.

21-32

Authors: G. N. Nikovskaya, K. V. Kalinichenko

Analysis of information on air-conditioning contaminated with heavy metals sludge municipal wastewater points to the actual ecological and chemical problem and its solution could be implemented within the framework of the biological process involving heterotrophic microorganisms. Information on the spread, toxicity, biochemistry, microbiology, colloidal and chemical properties of sludge sediments of municipal wastewater biological treatment is given in the review. These sediments contain vitamins, amino acids, organic matter, heavy metals (micro- and macroelements). Therefore the most rational approach to sludge wastes utilization is their use as an agricultural fertilizer after partial removal of heavy metals. Hence, the interaction of sludge components with heavy metals, modern methods of their removing from biocolloidal systems and biotechnologies of conversion of sludge wastes into fertilizer based on the enhancing of vital ability of sludge biocenoses are discussed.

43-51

Authors: D. M. Nozdrenko, K. I. Bogutska, Yu. I. Prylutskyy, U. Ritter, P. Scharff

Effect of pristine C60 fullerene aqueous colloid solution (C60FAS; 1 mg/kg dose) on the dynamics of fatigue processes in rat soleus muscle after ischemia-reperfusion injury using the tensiometric method was studied. Experiments were conducted during the first 5 h and for 5 days after ischemia. The changes in maximal strength of muscle contraction and its level of generation between the beginning and end of stimulated irritation after intravenous and intramuscular administration of C60FAS unmodified fullerene aqueous colloid solution were analyzed. The pronounced protective effect of this drug on the dynamics of skeletal muscle contraction was first determined. Protective effect of C60FAS unmodified fullerene aqueous colloid solution relative to changes in the levels of muscle contraction strength generation between the beginning and end of stimulated irritation was 15% in the first 5 h after ischemia and increased to 92% on the 5th day of the experiment. In such a case, the intravenous therapeutic administration of C60 fullerene aqueous colloid solution was the most optimal: the protective effect was 67% versus 49% under intramuscular administration. Thus, the development of biomedical nanotechnology with the application of pristine C60 unmodified fullerene as a strong antioxidant opens up new possibilities in prevention and treatment of ischemic injury in the skeletal muscles.

60-68

Authors: M. D. Kuchma, V. A. Shablii, V. M. Kyryk, A. N. Svitina, Yu. K. Prokopets, T. M. Indichenko, O. M. Tsupykov, L. L. Lukash, G. S . Lobyntseva

It have been shown that progenitor cells of cord blood, bone marrow and «mobilized» peripheral blood in semisolid mediums gave flat erythroid colonies. These colonies are able to form one or more red centers on the 14th day of cultivation and get a big size that evidence about high proliferative activity and resemble granulocyte, erythrocyte, monocyte/macrophage, megakaryocyte colony-forming units. However 92% of the cells of flat colonies express CD235. It shows that the colonies are erythroid, although colony morphology differs from burstoforming erythroid units and erythroid colony forming units. Their occurrence probability in methylcellulose-containing medium is 2,5%±1%, that is significantly lower than in agar- containing medium (58%±4,8%). Thus, we suggested that flat colonies should be counted separately or they should be ascribed as BFU-E.

74-80

Authors: V. V. Turov, T. V. Krupskaya, N. P. Galagan

The study of the process of melting water in lactose-glycerol-yolk kriomedium containing gametes bull, incorporated in the hydrophobic silica powder, which are adsorbed on the surface of fixed amounts of nonpolar hydrocarbon – n-decane was the aim of the work. The possibility of water polyassociates structuring with a solid surface of interfacial water and solubility of trifluoroethanoic acid in it have been studied. Thereat survival of the germ cell after contact with the surface was not analyzed. State of water in initial cryoprotective glycerol-lactose-yolk medium and hydrophobic nanosilica TS-100 containing n-decane additive adsorbed on its surface incorporated in a matrix was studied using low-temperature 1H-NMR spectroscopy method. It is shown that the solid matrix induces formation of 6–7 water molecules per each dean molecule at the interface, which do not take part in formation of hydrogen bonds, and a sharp radius decrease (from 100 to 20 nm) of ice crystals formed in cell suspension at its freezing. The results could give rise to safety improving of their cells at their cryopreservation and low temperature storage conditions by incorporating into a powder composite environment.

88-94

Authors: T. Yu. Trohimchuk, V. G. Serdyuk, L. O. Ganova, Y. I. Gorlov, S. V. Mihalap, N. Ya. Spivak

The problem of early diagnosis of human immunodeficiency virus remains relevant and can be solved by introduction the highly sensitive 4th generation ELISA test kits in the laboratory practice. The advantage of this analysis is simultaneous determination of serum antigen p24 HIV-1 and HIV-specific total antibody (IgG, IgM, IgA), that can detect the infection in the early stages of development. In Private Joint Stock Company «Scientific and Production Company Diaproph-Med» a modified version of the test kit of the 4th generation DIA-HIV-Ag/Ab hypersensitivity was developed, which uses a mixture of synthetic analogs of viral antigens in the immune-enzyme conjugates at two stages of reaction. Significant amplification of specific signal is achieved by introducing a streptavidin which is conjugated to horseradish peroxidase polymeric form. The analytical sensitivity of the test kit to identify the 1st International Standard WHO HIV p24 (NIBSC) was 0.78 IU/ml. The ability to test kit to detect early HIV infection has been examined for 9 seroconversion panels of sera (7 production ZeptoMetrix Corp. and 2 – SeraCare Life Sciences, USA) and is comparable to its peers, the leading manufacturers of test kits. The diagnostic characteristics of the test kit tested on standard panels, including dilution of serum of HIV- infected people, and not diluted sera with antibodies to HIV-1 (1 869 samples) and HIV-2 ( 41 sample). In these studies, the test kit detected HIV in a large ratio OD/cut off compared to similar commercial test kits for the 4th generation. The diagnostic specificity of the test kit DIA-HIV-Ag/Ab according to the results obtained in the study of blood donors from various transfusion services in Ukraine (93 788 researches) was 99.95%.

102-108

Authors: S. V. Stepaniuk, V. G. Naidenov, M. Y. Spivak

Results of the annual monitoring of epizooties indicate that highly pathogenic HPAI/H5N1 avian influenza widely circulated in Eurasian region. Over a period of 2010–2013 years more than 165 cases of outbreaks in 14 countries were found out. Ukraine became one of the first countries in Europe where in Autonomous Republic of Crimea in October 2005 outbreak of avian epizootic with HPAI/H5N1 was documented and until February 2008 more than 236,000 poultry were killed. Since then the question of monitoring of infected both migrating birds and poultry in places of cross contact in Ukraine remains of high priority. The test system is developed for identification and genotyping A H5N1 on three genes (M, H5 and N1) HPAI/H5N1 in real-time mode for polymerase chain reaction. Test kit capacity to detect HPAI/h5n1avian influenza virus and differentiate it from the other viral infection agents of birds and animals were studied by testing of HPAI/H5N1 virus isolated during mass infection outbreak in Crimea in 2005 and cultural specimens of other viral pathogens. It was established that the «DIA Real Avian Influenza» test kit was capable to detect RNA influenza A virus of high pathogenic H5N1 strains having high sensitivity (100% while RNA of the Crimean HPAI/H5N1 isolate studying) and specificity (100% while RNA viruses of Newcastle birds disease, fowl powershift, syndrome of drop in egg production and horse influenza studying).

26-33

Authors: V. M. Mokrosnop, E. K. Zolotareva

Microalgae are able to accumulate considerable amounts of tocopherols (up to 4 mg/g dry weight). The content of α-tocopherol to plant oils is low, whereas microalgae contain up to 97% of the tocochromanols that provides high bioactivity. The data about the content of tocopherols in eukaryotic microalgae Dunaliella tertiolecta, Nannochloropsis oculata, Isochrysis galbana, Euglena gracilis, Tetraselmis suecica, Diacronema vlkianum, as well as in the cyanobacterium Spirulina platensis are given in the paper. The largest amounts of tocopherols are synthesized by Euglena gracilis cells at mixotrophic cultivation. The level of tocopherols in microalgae depends on cultivation conditions. Two-stage biotech cultivation techniques, limiting nutrition in some biogenic elements, the introduction of exogenous carbon sources are used to increase the yield of tocopherol from microalgae. The approaches to the genetic transformation of plants leading to higher content of active vitamin E are rewieved as well.

46-53

Authors: N. V. Korotkevich, A. Ju. Labyntsev, D. V. Kolibo, S. V. Komisarenko

Heparin-binding EGF-like growth factor (HB-EGF) belongs to the epidermal growth factor receptor family and is synthesized as a transmembrane precursor proHB-EGF. Binding of sHB-EGF with receptors EGFR and HER-4 results in formation of ligand-receptor complexes and activation of signaling pathways and thereby promotes survival, proliferation and migration of cells. The aim of the study was to obtain the recombinant fluorescent derivatives of full-length soluble sHB-EGF – mCherry-sHB-EGF and truncated sHB-EGFΔ84-106 (without heparin-binding domain) – mCherry-sHB-EGFΔ84-106. The recombinant fluorescent derivatives may be used to investigate the sHB-EGF binding with receptors EGFR and HER-4, intracellular transportation of ligand-receptor complexes and the role of sHB-EGF heparin-binding domain in sHB-EGF biological activity. It was shown the ability of both fluorescent derivatives specifically bind to EGFR, to internalize in receptor-mediated pathway and to enhance the proliferation of 3T3 cells. Absence of heparin-binding domain in structure of mCherry-sHB-EGFΔ84-106 significantly did not effect on its ability to bind with receptor, but decreased twice its mitogenic activity. Thus, obtained fluorescent derivatives of sHB-EGF could be a convenient tool for investigation of molecular mechanisms of sHB-EGF biological activity and role of heparin-binding domain in these processes.

63-69

Authors: M. O. Fomina

The aim of work was to elucidate the mechanisms of bioaccumulation of copper leached from minerals by fungus Aspergillus niger with great bioremedial potential due to its ability to produce chelating metabolites and transform toxic metals and minerals. The special attention was paid to the chemical speciation of copper bioaccumulated within fungal colony in the process of fungal transformation of copper-containing minerals. Chemical speciation of copper within different parts of the fungal colony was studied using solid-state chemistry methods such as synchrotron-based X-ray absorption spectroscopy providing information about the oxidation state of the target element, and its coordination environment. The analysis of the obtained X-ray absorption spectroscopy spectra was carried out using Fourier transforms of Extended X-ray Absorption Fine Structure regions, which correspond to the oscillating part of the spectrum to the right of the absorption edge. Results of this study showed that fungus A. niger was involved in the process of solubilization of copper-containing minerals resulted in leaching of mobile copper and its further immobilization by fungal biomass with variable coordination of accumulated copper within fungal colony which depended on the age and physiological/reproductive state of fungal mycelium. X-ray absorption spectroscopy data demonstrated that copper accumulated within outer zone of fungal colony with immature vegetative mycelium was coordinated with sulphur–containing ligands, in contrast to copper coordination with phosphate ligands within mature mycelium with profuse conidia in the central zone of the colony. The findings of this study not only broaden our understanding of the biogeochemical role of fungi but can also be used in the development of various fungal-based biometallurgy technologies such as bioremediation, bioaccumulation and bioleaching and in the assessment of their reliability. The main conclusion is that coordination environment of copper bioaccumulated within fungal biomass via the process of transformation of copper minerals is heterogeneous varying from sulphydryl to phosphate.

79-85

Authors: H. V. Dziak, S. M. Shulga, M. Adab, O. L. Drozdov, I. S. Glukh

Dry lecithin, which is a mixture of polar phospholipids, neutral lipids, free fatty acids, glycolipids, carbohydrates, and small amounts of moisture, is a promising object for biologic-hepatoprotectors creation. One of its pharmacological activity displays is its influence on serum lipids, in particular transport forms of these lipids. The influence of dry soy lecithin and sunflower on hyperlipoproteinemia ratio and other lipid disorders is studied. It is shown that low-fat dry soybean lecithin showed hypocholesterolemic activity against all studied forms of serum cholesterol. Nonfat dry sunflower lecithin had similar but somewhat less prominent effect. However reduced concentration of high density lipoprotein cholesterol under sunflower lecithin developed right before soya lecithin. Both lecithin prevented the development of dyslipidemia induced carbon tetrachloride.

92-105

Authors: O. A. Akinloye, E. A. Balogun, S. O. Omotainse, O. O.Adeleye

The study was aimed to investigate some untoward effects that could be associated with the use of P. nitida as hypoglycemic agent using some biochemical and histological evidences. The antioxidant property of the plant was determined by using 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity. Biochemical studies in plasma using determining the testes such as blood glucose, alanine and aspartate aminotransferases, gamma glutamyl transferase activities, electrolytes (sodium, potassium and bicarbonate, lipid peroxidation levels, haematological parameters (red blood cell and whole blood cell, platelets, and lymphocyte counts), blood glucose level, lipid profile, and also liver and kidney function tests were performed. Histopathological examinations of the liver, kidney and pancreas were done following the standard Heamatoxylin and Eosin staining method. Methanol extract of the seeds has the highest antioxidant level (36.73%), indicating higher free radical scavenging activity; followed by aqueous extract (19.36%) and coconut water extract (4.09%). There was significant reduction (p<0.05) in blood glucose of all the treated rats at the end of the experiment (ranging from 55.59% to 41.66%). Significant increase (p<0.05) in body weights of the treated rats were also observed at the end of the treatment (ranging from 9.26% to 38.89%). There was a significant (p<0.05) increase in the hematological parameters in all the extract treated groups. There was also significant decrease (p<0.05) in the lipid profiles of the treated groups. Plasma studied enzymes activities decreased in all treated groups. Ionoregulatory disturbances observed included hyperkalemia and hypernatremia in all the treated groups but were reduced significantly (p<0.05) at the end of the treatment. Urea and bicarbonate concentrations and also of lipid peroxidation level decreased significantly in all the groups. The histopathological studies revealed that the extracts were unable to ameliorate some observable pathologic conditions associated with induced diabetic tissues. Although, diabetes mellitus have been reported to be associated with varied histological changes in different organs, in this study, histological examinations of the pancreas of the treated and untreated groups showed varying degree of degenerations but the extent of severity in the lesions were more pronounced in the extract treated groups. In this relation the obtained results of this study which revealed the hypoglycemic and antioxidant potentials of Picralima nitida seed extracts for the treatment of diabetes mellitus should be taken with caution in administering the P.nitida seed extract as an hypoglycemic agent.

114-117

Authors: E. A. Raksha-Slusareva1, V. A. Krul1, А. А. Slusarev

When oil and biodiesel are extracted from rapeseed, secondary derived products are formed, which are not used effectively at the moment. The article deals with the problems of possible their use in food industry. During food product preparation for special dietary consumption we used electrophysical (processing by hydroelectropulse) and physical (drying, grinding, steam treatment) processing of raw materials. Through the developed technology for rapeseed cake processing, we received raw materials suitable for use in food industry. On the basis of these raw materials, the «Nutrition product for special dietary consumption “Ripakovyi”» was developed. It is a part of rape seed meal obtained from the seeds with low content of glucosinolates and erucic acid processed by hydroelectropulse dried in the cabinet oven or in the convective dryer, crushed and disinfected based on a developed soft technology for biologically active substances conservation. The production of this product solves the problem of rational utilization of rapeseed meal and diversification of foods for special dietary consumption.