Biotechnologia Acta

9-18

Authors: V. I. Nazarenko, O. P. Demchenko

This review addresses the analysis of properties and applications of fluorescent nanodiamonds. They are carbon nanostructures with atomic arrangement of a diamond and carry all its properties, including record — high density, rigidity and refraction index. They are of almost spherical shape, and their small size (~4–10 nm) creates substantial surface area that can be used for absorption of different compounds including drugs. Their surface is formed by different chemical groups (hydroxyls, carboxyls, etc.) exhibits also chemical reactivity that allows different types of modifications. This opens innumerable possibilities for constructing different functional nanomaterials. The technologies have been developed for making these nanodiamonds fluorescent. Particularly, these properties are achieved by radioactive treatment with the formation of N–V impurities. These particles absorb and emit light in convenient for observation visible range of spectrum. They do not photobleach, which is very attractive for fluorescent microscopy of the cell. And, finally, these nanoparticles do not display toxicity on the cellular or whole — body level, and because of their biocompatibility they can be used in vivo as contrast agents and drug carriers. It is expected that future biotechnological applications of these nanoparticles will be connected with the creation of nanocomposites that combine multiple useful functions.

41-48

Authors: M. V. Grigoreva

Biodegradable polyurethanes attract interest of those developing composite materials for biomedical applications. One of their features is their ability to serve as carriers, or matrixes, for medicines and other bioactive compounds to produce a therapeutic effect in body through targeted and/or prolonged delivery of these compounds in the process of their controlled release from matrix. The review presents polyurethane composites as matrices for a number of drugs. The relation between structure of the composites and their degradability both in vitro and in vivo and the dependence of drug release kinetics on physicochemical properties of polyurethane matrix are highlighted. The release of drugs (cefazolin, naltrexone and piroxicam) from the composites based on cross-linked polyurethanes (synthesized from laprols, Mw between 1,500 and 2,000 Da and toluylene diisocyanate) demonstrated more or less the same pattern (about 10 days in vitro and three to five days in vivo). In contrast, the composites with dioxydine based on a linear polyurethanes (synthesized from oligotetramethilene glycol, Mw 1,000 Da, diphenylmethane-4,4’-diisocyanate and 1,4-butanediol) retained their antimicrobial activity at least 30 days. They also showed a significantly higher breaking strength as compared to that of the composites based on cross-linked polyurethanes.

62-78

Authors: V. D. Naumenko, B. V. Sorochinsky, V. I. Kolychev

Biological properties, chemical structures and biosynthesis pathways of plant isoflavones, especially soybean isoflavones (daidzein, genistein and glycitein) are reviewed. The structures of isoflavones, and their aglicone and glucosides (glycosides) forms as well as isoflavone biosynthesis pathways are described. General information about the advanced methods for the detection of isoflavones and their conjugates are considered. The importance of the profiling of isoflavones, flavonoids and their conjugates by means of analytical tools and methods to dissolve some questions in biology and medicine is discussed. The review provides data on the major isoflavone content in some vegetable crops and in the tissues of different soybean varieties. Health benefits and treatment or preventive properties of isoflavones for cancer, cardiovascular, endocrine diseases and metabolic disorders are highlighted. The mechanisms that may explain their positive biological effects are considered. The information on the application of advanced technologies to create new plant forms producing isoflavonoids with a predicted level of isoflavones, which is the most favorable for the treatment is given. The possibilities to use the metabolic engineering for the increasing of accumulation and synthesis of isoflavones at the non-legume crops such as tobacco, Arabidopsis and maize are considered. The examples how the plant tissues, which are not naturally produced of the isoflavones, can obtain potential for the synthesis of biologically active compounds via inducing of the activity of the introduced enzyme isoflavon synthase, are given. Specific biochemical pathways for increasing the synthesis of isoflavone genistein in Arabidopsis thaliana tissues are discussed. It is concluded that plant genetic engineering which is focused on modification of the secondary metabolites contain in plant tissues, enables to create the new crop varieties with improved agronomic properties and nutritional characteristics.

87-94

Authors: О. S. Gojster, W. Е. Kriwenchuk, G. O. Chmelnitsky, О. Н. Minchenko

The aim of this work was the development of T-2 toxin immunedetection method using biosensor approaches. For this aim the conjugates of T-2 toxin with proteins were obtained and used for antibody preparation. The specific properties of obtained antibodies were studied by immunochemical methods. It was shown that carrier enzyme plays important role in realization of immunogenic properties of T-2 hapten. The detection time of IgG fraction as well as antiserum binding activity by immunoenzyme and immunosensor analyses is 10–15 minutes. Two variants of T-2 toxin determination in competitive mode of immunochemical analysis were suggested, i. e. ELISA and immunosensor method (based on surface plasmon resonance phenomenon) with detection from 10 ng/ml and 1 ng/ml, correspondingly. These methods can be applied for detection of T-2 toxin in the food and fodder in concentration from 1 up to 500 ng/ml.

100-107

Authors: S. V. Malysheva, N. M. Bilko, J. Hescheler

The search for effective and safe methods to generate induced pluripotent stem cells is especially urgent. In the paper murine embryonic fibro blasts were reprogrammed towards actively proliferating colonies with typical induced pluripotent stem cells morphology by means of Sleeping beauty transposon-based vector system. The obtained clones were checked for the expression of various pluripotency markers: alkaline phosphatase, Oct4 and Sox2 genes, SSEA-1 expression in various clones was evaluated. Also the reactivation of endogenous pluripotency factors Nanog and Rex1 was indicated. The data obtained is analyzed and compared to the established pluripotent stem cell line. It is shown that somatic cells are reprogrammed towards pluripotency by means of Sleeping beauty transposon system. Therefore, the system is a new perspective biotechnological tool to generate pluripotent cells.

115-121

Authors: О. Yu. Rogulska, О. B. Revenko, Yu. O. Petrenko, H. Ehrlich, О. Yu. Petrenko

Development of the new types of tissue engineered structures is one of the promising trends of current biotechnology. The study was directed to the assessment of prospects for the application of chitin-based skeletons derived from marine sponges of Aplysinidae family (Aplysina fulva and Aplysina aerophoba) for creation of bioengineered constructs based on human mesenchymal stromal cells. After cleaning and demineralization procedures, sponge skeletons appeared as three-dimensional macroporous matrices formed by intersecting chitin fibrils. After seeding into chitin-based matrices the cells were attached to the surface of the fibrils and were able to spread and proliferate. Mesenchymal stromal cells within Aplysina fulva differentiated into osteogenic and adipogenic directions under the influence of appropriate inductors. Demineralized skeletons derived from marine sponges of Aplysinidae family could be used as scaffolds for mesenchymal stromal cells which provides new opportunities for the creation of adipose and bone tissue engineered structures.

131-136

Authors: O. M. Magomedov, A. F. Levytsky, D. V. Golovatiuk, A. N. Velikyy, T. A. Polishchuk

The results of the study of diagnostically important metabolism parameters in patients with bone cysts in different stages of the disease are presented. It is shown that an increase activity of protein banding collagenase, alkaline phosphatase and also of hydroxyproline, glycosaminoglycans contents due to lower levels of calcium and inorganic phosphate levels increase in blood serum are expressed in a stage osteolysis than the step of separating. Decreasing the amount of glycosaminoglycans and collagen in bone indicates an intensification of catabolic processes in the connective tissue matrix. Diagnostically important indicators of the degree of disturbance of bone metabolism are the level of collagen, proteoglycans and activity of marker enzymes — collagenase and alkaline phosphatase. Based on the evaluation of sensitivity, specificity and diagnostic efficiency of the obtained results, we can recommend the threshold values of the investigated parameters of basic organic components and mineral metabolism of bone for the differential diagnosis of stages of bone cysts in children, which will serve as a basis for the development of appropriate diagnostic tests.

143-149

Authors: О. V. Кyrychenkо, S. O. Gryniuk

The growth-regulatory activity of the complex algae extract from Spirogira sp. at pre-so wing treatment of soybean seeds was studied in green-house and field experiments. It was shown that phytoextract has stimulated seeds germination on 12%, plants growth on 11–37%, soybean productivity increased on 6–27% as well as activated the development and functional ability of rhizospheric nitrogen-fixing microorganisms. The quantity of oligoazotrophes increased in 1,5–6,3 times, nitrogenase activity in 1,5–1,7 times. The possibility aspects of growth-regulatory activity of the algae extract in both plants and rhizospheric microorganisms is under discussion. Our results have confirmed the perspectives of practical use of the biological activity substances from the algae at soybean growing in order to increase plants productivity and improve microbiological indexes of soil.