[Effect of alkaloids from celandine on calcium accumulation and oxidative phosphorylation in mitochondria depending on their DNA intercalating properties].

The effect of principal alkaloids (sanguinarine, chelerythrine, coptisine, chelidonine) of greater celandine Chelidonium majus L., as well as the alkaloids from Colchicum autumnale L. (colchicine and colchamine) on calcium accumulation and oxidative phosphorylation in rat liver mitochondria has been studied. The obtained data were compared with DNA intercalating properties of alkaloids detected by the method of thermodenaturation (DNA melting curve plots). It was found that chelerythrine and sanguinarine blocked absorption and accumulation of calcium cations and inhibited oxidative phosphorylation, while the coptisine significantly diminished those indices. Chelidonine, colchicines and colchamine had no influence on the studied characteristics. The effect of alkaloids upon mitochondria functional state correlated tightly with their DNA intercalating properties: chelerythrine and sanguinarine were strong intercalators, while coptisine was a weak one, and chelidonine, colchicine and colchamine did not interact with DNA and caused no changes in its melting point. Correlation coefficient between the intercalating properties of alkaloids and their inhibition of calcium accumulation was 0.89, and with their oxidative phosphorylation inhibition - 0.93. It is suggested that the effect of studied alkaloids upon functional properties of mitochondria can be mediated by mtDNA.

[Comet assay of DNA fragmentation: modification of silver staining for obtaining permanent preparations].

Modification of comet analysis is proposed for obtaining permanent preparations by DNA staining with silver compounds. The sensitivity of staining is similar to that observed at the treatment by ethidium bromide and other fluorochromes. The advantages of the method are stability of slides and possibility of their reinvestigation by light microscopy. The method does not need expensive fluorescent microscope and lacks contacting with carcinogenic compounds and UV light irradiation.

[Carbohydrate-binding proteins--lectins and glycosidases in plants of Anthurium genus]. / Doslidzhennia vuhlevodzv'iazuiuchykh bilkiv--lektyniv ta hlikozydaz u roslyn rodu anturium (Anthurium).

The content of lectins and activity of glycosidases have been estimated in seeds and vegetative organs of 5 strains of plants of Anthurium genus. In seeds of all the investigated strains lectins were detected with the selectivity toward the N-acetyl-galactosamine (minimal inhibitory concentration of sugar was 0.1-0.2 mM) and an anti-A blood group specificity. Lectins of Anthuriums selectively bound O-type glycosidic chains and revealed high affinity toward mucins (salivary or ovary cysts origin). Lectins were not detected in vegetative parts of Anthuriums. In seeds of plants the following glycosidases were detected in the diminishing activity order: alpha-galactosidase, alpha-mannosidase, beta-galactosidase, beta-glucosaminidase.

[Production of lectin from jack fruit (Artocarpus integrifolia) seeds and its interaction with carbohydrates and glycoproteins]. / Oderzhannia lektynu iz nasinnia khlibnoho dereva (Artocarpus integrifoli) i ioho vzaiemodiia z vuglevodamy ta glikoproteïnamy.

A method is developed to obtain lectin from jack fruit (Artocarpus integrifolia) seeds using an affinity chromatography on a sorbent prepared from the egg white. The minimum agglutination concentration of human erythrocytes is 80 ng/ml, the molecular weight of the preparation is about 39 kDa, it contains 1.8% of neutral hexoses and 3.1% of hexosamines. PAAG electrophoresis in the alkali system has revealed several molecular forms of lectin isolated by preparative electrophoresis, their properties are investigated. SDS-PAAG electrophoresis has revealed several types of polypeptide chains among which two chains (12 and 14 kDa) are predominant. Lectin possesses affinity to galactosides (not to free galactose) and N-acetylgalactosamine and interacts with O-glycans with high affinity. The preparation has mitogenic activity in optimal concentration 50 micrograms/ml.

[Electroelution of protein fractions from a polyacrylamide gel]. / Elektroeliuiuvannia bilkovykh fraktsii z poliakrylamidnogo geliu.

A method for electroelution of protein fractions from polyacrylamide gel and device for performing such a process have been developed. The application of two tris-glycine buffers with the low and high ionic strength, pH 9.0-9.2 provides a concentration of protein simultaneously to extraction from the gel. The duration of elution is in the range of 1-3 hours and depends on the protein mobility. The effectiveness of the system is demonstrated for disc-electrophoretic separation and electrophoresis in slab gel in the presence of SDS. The maximal amount of pure protein fraction obtained is about 4.5-5.0 mg. The method may be useful especially for the fractionation of limited quantities of protein samples.

The toxicity and antitumor activity of three individual fractions of lectins from Ricinus communis seeds.

Lectins from Ricinus communis seeds (RCL) have been resolved into three electrophoretically distinct fractions (alpha-RCL, beta-RCL, psi-RCL) by ion exchange chromatography on Watman CM-32 cellulose. The toxicity to mice and antitumor activity against murine lymphoma NK/Ly of pure fractions were compared. Optimal effect was achieved with alpha-RCL, Which possessed the lowest toxicity (1000 micron g/kg) and significant antitumor activity (63% inhibition of tumor growth).