M. Lupak's research while affiliated with Ivan Franko National University of Lviv and other places

The article describes the effect of alkaloid-free fraction of the Galega officinalis extract (AFFGE) on the aggregation ability of immunocompetent blood cells, as well as on the process of actin polymerization and structural rearrangements among sialylated glycoconjugates of the peripheral blood leukocyte membranes of rats in the norm and under experimental diabetes mellitus (EDM) conditions. The flow cytometry method (using phalloidin labelled with fluorescent tetramethyl rhodamine-5-isothiocyanate (TRITC)) and the western blot analysis have allowed us to detect an increase in the rat leukocyte F-actin content in the event of diabetes mellitus, which indicated changes in the structural and functional properties of the leukocytes and their preactivation phase. A quantitative analysis of the total polymerized actin pool redistribution between its constituent fraction (represented by cytoskeletal filaments) and short actin filaments has shown that, against an increase in the total F-actin level, the number of actin filaments of the cytoskeleton decreased and the content of short actin filaments increased in leukocytes of animals with EDM. The use of sialylated lectins has allowed a conclusion to be made on the study of the pathology that the number of exposed oligosaccharide determinants on leukocyte membrane, the structure of which contained N-acetyl-β-D-glucosamine and sialic acid residues, increased, whereas the number of sialic acid-containing surface glycoconjugates bound to subterminal galactose residues by α2→3 and α2→6-glycoside bonds decreased. The administration of AFFGE to diabetic animals led to an increase in the content of F-actin and short filaments of the leukocyte cytoskeleton and a reduction in the lectin-induced leukocyte aggregation. The correction effect of the studied extract on the functional state of leukocytes can be realized through the action on the processes underlying the formation of the actin cytoskeletal elements and due to the quantitative redistribution of leukocyte membrane glycoconjugates with different structures of carbohydrate determinants, such as, due to a decrease in the exposure of N-acetyl-β-D-glucosamine residues and an increase in the exposure of sialic acids bound to subterminal galactose residues by α2→3 and α2→6-glycoside bonds.

The article contains data on the influence of biogenic surfactants synthesized by bacteria Pseudomonas sp. PS-17 on the stability of emulsions based on alkaloid-free fraction isolated from Galega officinalis L. extract. The opti­mum concentrations of biogenic surfactants to stabilize investigated emulsionswere experimentally chosen. The chemical composition of alkaloid-free fractions from Galega officinalis L. extract before and after stabilization were investigated. The resulting substance is easier to administer to animals through a tube, which not only improves the accuracy of the dosage, but also increases its bioavailability. Reduction of glycosylated hemoglobin at 13.4 % and glucose content to the physiological values in the rats blood with experimental diabetes mellitus by administration within 14 days stabilized emulsions compared with the original emulsion shows that emulsion stabilized by the biogenic surfactant based on alkaloid-free fraction isolated from Galega officinalis an extract provides effective hyperglycemia compensation in rats under experimental diabetes mellitus, compared with the original mixture, which may be caused by changes in the proportion of biologically active substances in the extract while stabilizing and its better bioavailability.

The article contains the data of Galega officinalis extract (GOE) nonalkaloid-containing fraction influence on the structural and functional state of leukocytes under the condition of experimental diabetes mellitus type 1 (EDM). Immunohistochemical method has shown the quantitative redistribution of leukocytes containing proapoptotic protein p53 and PARylated [poly(ADP-ribosyl) ated] proteins under EDM conditions and admission of the tested extract. GOE administration to animals with EDM leads to the reduction in the number of cells with positive and strongly positive reaction to the p53 protein and PARylated proteins content, indicating its depressing effect on apoptosis of immune cells. 2013