Biotechnologia Acta

23-36

Authors: Pirhanov G.G., Zhernossekov D.D.

Sinorhizobium meliloti is a Gram-negative soil nitrogen-fixing bacterium that increases the yield of legumes. There is information in the literature about the complete genome sequence of this bacterium, in addition, the polysaccharide composition of the biofilm, which is actively involved in nitrogen fixation, has been studied. The well-known nucleotide sequence, as well as the genetic and biochemical features of S. meliloti make this organism an ideal model for biotechnological research. The purpose of this work was to analyze the current data provided in the literature on the symbiotic interaction of Sinorhizobium meliloti with the host plant, and to characterize the main directions of the use of this bacterium in agriculture, bioremediation and medicine.

44-52

Authors: O. K. Gulevskyy, Yu. S. Akhatova

One of the modern transfusiology tasks is to preserve the properties of erythrocytes after hypothermic storage. Worsening of their functional state due to the storage leads to deterioration of the transfusion medium quality and a number of clinical problems. Plausible protective effects of a human cord blood low-molecular fraction (CBF) and the Actovegin drug were evaluated in the present study in order to use them as components of recovery medium. Aim. The purpose of our study was to investigate the CBF and the Actovegin drug effect on erythrocytes morphology, energy balance, and oxygen transport function of erythrocytes after their hypothermic storage. Materials and methods. In the research erythrocyte mass from human donor blood was used, which was stored in hypothermia for 7–21 days. Every 7 days, the CBF or Actovegin (final concentration 0.6 mg/ml) were added to aliquots of erythrocyte mass and incubated for 1 h at 37 °C, followed by further studies. CBF was obtained from human cord blood by ultrafiltration and lyophilization. The erythrocytes morphology was assessed using light microscopy. The content of ATP and 2.3-DPG was determined by the amount of inorganic phosphorus and the photoelectrocalorimetric method. The indicators of erythrocytes oxygen transport function (saturation, oxygen and carbon dioxide tension) were assessed using an analyzer of blood gases and electrolytes IL GEM Premier - 3000. The ratio of hemoglobin forms was studied by photometry. Results. It was shown that both CBF and the Actovegin drug promoted to restore the morpho-functional characteristics of erythrocytes after 21 days of their storage at 24 °C. This was indicative as an increase in the normocytes number increase and restoration of oxygen tension, saturation, ATP and 2,3-DFG content, and normalization of the hemoglobin forms ratio. It was suggested that the mechanisms of the compared tested substances action were associated with ATP synthesis activation and 2,3-DFG formation. Conclusions. The use of media with containing CBF or the Actovegin drug enabled to restore effectively the properties of erythrocytes disturbed during their prolonged storage.

60-70

Authors: N. Kobylinska, T. Bohdanovych, V. Duplij, I. Pashchenko, N. Matvieieva

Aim. The profiles of polyphenolic phytochemicals in extracts of “hairy” roots of Artemisia tilesii Ledeb. and Artemisia annua L. were studied. Analytical separation and quantification of main components in extracts were evaluated. Methods. “hairy” roots were grown in vitro on Murashige and Skoog medium. High-performance chromatography coupled with different types of detection (photo diode array detection (DAD) and electrospray ionization with ultra-high resolution Qq-Time-of-Flight mass spectrometry) was used to identify and quantify the main biologically active components in ethanol extracts of “hairy” roots. Results. The amount of flavonoids was 94.71–144.33 mg RE/g DW and 33.52–78.00 mg RE/g DW in “hairy” roots of A. annua and A. tilesii, respectively. In most samples of “hairy” roots, the amount of flavonoids was higher than the content in the control plant roots. The presence of Apigenin (0.168 ± 0.003 mg/L and 0.178 ± 0.006 mg/L), Quercetin (0.282 ± 0.005 mg/L and 0.174 ± 0.005 mg/L) in the extracts of A. annua and A. tilesii was shown by reverse-phase HPLC-DAD method. Chlorogenic acid, Kaempferol, and other flavonoids were detected. Conclusions. The developed HPLC-DAD method demonstrated the high percentage of recovery, low limit of detection and quantification (9,11 ng/ml ≤ LOQ ≤16,51 ng/ml), accuracy and correctness. Thus, the method is suitable for the simultaneous quantification of phenolic acids and flavonoids in various plant extracts with short time and high efficiency.