[The Redox-Catalytic Properties of Cobalamins].

Vitamin B12, or cobalamin, is essential for normal body function and is used in the therapies of different diseases. Vitamin B12 has anti-inflammatory and antioxidant properties that can play an important role in the prevention of some diseases. On the other hand, it has been reported that vitamin B12 in combination with such reducing agents as ascorbate (vitamin C) and thiols showed prooxidant activity. This review provides information on the roles of vitamin B12 in diseases accompanied by inflammation and oxidative stress and the effects of vitamin B12 administrated alone and in combinations with different reducing agents such as ascorbate and thiols on oxidative stress. In addition, the mechanisms of prooxidant actions of combinations of vitamin B12 with these reducing agents depending on the form of vitamin B12 (hydroxocobalamin and cyanocobalamin) are discussed. Understanding the mechanisms of prooxidant action of vitamin B12 is necessary for developing strategies for therapeutic administration of vitamin B12.

[Partitioning of taxifolin-iron ions complexes in octanol-water system].

The composition of taxifolin-iron ions complexes in an octanol-water biphasic system was studied using the method of absorption spectrophotometry. It was found that at pH 5.0 in an aqueous biphasic system the complex of [Tf2 x Fe x (OH)k(H2O)8-k] is present, but at pH 7.0 and 9.0 the complexes of [Tf2 x Fe x (OH)k(H2O)2-k] and [Tf x Fe x OH)k(H2O)4-k] are predominantly observed. The formation of a stable [Tf3 x Fe] complex occurred in octanol phase. The charged iron ion of this complex is surrounded by taxifolin molecules, which shield the iron ion from lipophilic solvent. During transition from water to octanol phase the changes of the composition of complexes are accompanied by reciprocal changes in portion of taxifolin and iron ions in these phases. It was shown that the portion of taxifolin in aqueous solution in the presence of iron ions is increased at high pH values, and the portion of iron ions is minimal at pH 7.0. In addition, the parameters of solubility limits of taxifolin-iron ions complexes in an aqueous solution were determined. The data obtained gain a better understanding of the role of complexation of polyphenol with metal of variable valency in passive transport of flavonoids and metal ions across lipid membranes.

[Effect of the liposomal form of taxifolin complexes with metals of variable valence on skin regeneration after chemical burn].

The effect of the liposomal form formed by taxifolin and metals of variable valency was investigated. It was shown that the application of preparations based on the free flavonoid and its complexes with Fe(II/III) and Cu(II) ions after chemical burns results in a more effective skin regeneration and the repair of hair follicles and sebaceous glands. A tendency for a more effective wound healing after the applications of taxifolin-Cu(II) and taxifolin-Fe(III) liposomal complexes versus control was observed. It was assumed that the mechanism of action of these preparations is based on the oxidative polymerization and conjugation of the flavonoid, which results in the utilization of toxic metabolites and lipid peroxidation products.

Skin regeneration after chemical burn under the effect of taxifolin-based preparations.

We studied the effect of liposome preparation containing taxifolin oligomers and taxifolin conjugates with carbonyl compounds on skin regeneration after chemical burn. The preparations containing flavonoid conjugates intensified regeneration processes and reparation of hair follicles and sebaceous glands after chemical burn. The preparation based on taxifolin conjugate with acetaldehyde was most effective; its activity was comparable with that of the wound-healing preparation Olasol. Taxifolin conjugates with carbonyl compounds can be used for the creation of combined wound- and burn-healing preparations.

[Cytoskeletal regulation of the cellular function by dopamine].

The interaction of dopamine with model membranes, isolated G-actin, and living cells, such as Mauthner neurons and fibroblast-like BHK-21 cells has been studied. It was found that in vitro dopamine passes through the phospholipid membrane and directly polymerizes G-actin due to incorporation into threads as their integral part. In in vivo conditions, it penetrates inside the cell and induces the appearance of a network of actin filaments in loci rich in globular actin. The data suggest that there exists a mechanism of dopamine interaction with living cells, which is based on direct polymerization of cytosolic G-actin as its cellular target. The reorganization of the actin cytoskeleton leads to changes in the morphofunctional status of cells.

[Cytochemical and ultrastructural characteristic of BHK-21 cells exposed to dopamine].

BHK-21 cells were incubated in a medium containing dopamine (DA) and then their catecholamine content evaluated by using the Falck cytochemical method. The significant intensification of cell fluorescence as compared to that one in control preparations was detected; this effect was proportional to DA concentration and exposure duration and was more pronounced in cells in suspension than in those attached to the substrate. Simultaneous ultrastructural investigation has shown that an increased intensity of the cytoplasm fluorescence correlated with the appearance of the dense network of fibrils that were morphologically identified as F-actin microfilaments. Prior blockade of dopaminergic receptors by haloperidol did not change the following DA effect both on the fluorescence intensity and cell ultrastructure. The data obtained suggest that DA chronically acting on the living cells was able to penetrate into the cytoplasm, causing actin polymerization and incorporating into the newly formed actin cytoskeleton. Structurally, this may be manifested by cytoskeleton and its derivative hypertrophy, that could have a substantial effect on general morphology of the cell.

[Influence of Biginelli pyrimidine on production reactive oxygen species by polymorphonuclear leukocytes].

The effect of 6 synthetic Biginelli pyrimidines on production of reactive oxygen species by polymorphonuclear neutrophils was investigated. It has been shown by method of luminol-dependent chemiluminescence that test compounds in a concentrations of 10-100 microM stimulate production of reactive oxygen species. An increase in the reactive oxygen species production by stimulated neutrophils in the presence of 10 microM 1-(3, 4-dimetoxyphenyl)ethyl-4(alkyl/aryl) substituted Biginelli pyrimidines was 50-90 %. An increase in the priming effect of Biginelli pyrimidines on reactive oxygen species production by neutrophils was noted in the case of replacement of phuryl radical by phenyl and alkyl radicals at C(4) pyrimidine cycle and in the case of replacement benzyl radical at N(1) by 3, 4-dimetoxyphenyl radical. It has been revealed a high inhibitory effect of 1-(3, 4-dimetoxyphenylethyl)-4(alkyl/aryl) substituted Biginelli pyrimidine in concentration 0.01-0.10 microM. It has been found that high concentration of ethyl-1-[2-(3, 4-dimethoxyphenyl)ethyl]-6- methyl-4-phenyl-2-thioxo-1, 2, 3, 4-tetrahydropyrimidine -5-carboxylate (1 mM and more) is able to cause respiratory burst of neutrophils without additional stimulation.

Effect of dopamine on viability of BHK-21 cells.

We studied the effects of dopamine added to culture medium on survival of floating or adherent BHK-21 cells differing by organization of actin cytoskeleton. The viability of floating cells more drastically decreased with increasing dopamine concentration and duration of exposure than that of adherent cells. The cells worse adhered to the substrate and formed a monolayer. The formed monolayer degrades, cell borders become blurred, cells, polygonal in the control, are rounded. Preliminary blockade of dopamine receptors with haloperidol, inessential for cell survival and morphology, does not prevent the destructive effect of dopamine on the cells. Ultrastructural study revealed increased density of filamentous actin threads in deep compartments of cell cytoplasm after dopamine treatment, this increase being more pronounced in cells grown in suspension. Bearing in mind the polymerizing effect of dopamine on globular actin in vitro and the fact that the content of this protein in floating cells is higher than in adherent cells, we can conclude that the decrease in viability of BHK-21 cells is caused by interaction of dopamine with cytoplasmic globular actin.

[The influence of dopamine on the ultrastructure of BHK-21 cells].

The influence of dopamine on the haloperidol of BHK-21 cells being in suspension or attached to substrate was investigated. It was shown that the ultrastructural changes affected mainly the cellular loci enriched by the cytoskeleton actin such as intercellular desmosome-like contacts, microvilli and cortical layer or mesh just beneath the plasmatic membrane. The desmosome-like contacts were hypertrophied, their electron density was increased and fibrilar bridges appeared in specialized contacts. Many microvilli fused with each other and with plasma membrane of the neighboring cells, or, on the contrary, split up. Frequently, the membrane surface between microvilli and particularly their apical parts was seen to be pierced by thin thread, morphologically similar to actin filaments. The cytoplasmatic matrix onto ultrathin sections had blotched appearance and at the ultrastructural level was represented by numerous randomly oriented actin filaments. The effect of dopamine was more pronounced in the BHK-21 cells when being in suspension than in attached to the substrate ones, which presumably occurred due to known lesser differentiation of the cytoskeleton in the formers. Finally, it was established that the preliminary blockade of cellular D2 receptors with haloperidol neither affected the ultrastructure of BHK-21 cells nor prevented the following effect of dopamine. The data obtained suggest the direct interactions of dopamine with the actin cytoskeleton.

[Interaction of dopamine with artificial phospholipid membranes].

The interaction of dopamine (DA) with phospholipid membranes has been investigated. The membrane current in planar bilipid membrane (BLM) modified by amphotericin B in voltage clamp conditions under alternating polarity was shown to symmetrically increase 1.2 times when DA was added outside the BLM. This implies a uniform change of charge on each membrane surface and hence the diffusion of DA within the BLM and its exposure on the internal side. The appearance of single threads and bundles of filaments within the internal liposomal cavities was observed in the ultrastructure of suspended thin-walled liposomes filled with globular actin after the introduction of DA into external solution. This reshaped liposomes into rod-like, spindle-shaped or angular structures. Actin serves as a marker for DA due to its property to polymerize itself under the influence of DA. Thus, the structural reorganization of liposomes manifests the presence of DA inside them and the induction of actin polymerization.

Changes in the ultrastructure and function of goldfish Mauthner neurons in the presence of 3,4-dihydro-2(1H)-pyrimidinethione.

The behavioral effects of 3,4-dihydro-2(1H)-pyrimidinethione (DPT, a pyrimidine derivative), which is used as a test system for detecting tumor growth, on the ultrastructure and function of Mauthner neurons (MN), were studied in goldfish. Application of DPT to MN was found to lead to increased resistance of neurons to exhaustive stimulation, which was accompanied by increases in the sizes of actin-containing membrane desmosome-like contacts, along with the formation of bundles of actin stress fibers; these effects are similar to those previously reported with dopamine. The similarity of the morphofunctional changes in MN on exposure to an artificial chemical substance for which there are no membrane receptors and dopamine itself suggests that they have trophic effects on the stabilization and polymerization of cytoskeletal actin due to direct penetration into postsynaptic neurons.

[Changes of the goldfish Mauthner neuron ultrastructure and function under the influence of 3,4-dihydro-2(1H)-pyrimidinethione].

The effects of pyrimidine derivative 3,4-dihydro-2(1H)-pyrimidinethione, (DPT) used as a test-system for detection of tumor growth, on the goldfish Mauthner neurons (MN) ultrastructure and function, as manifested in behavioral changes, were studied. The results of investigations demonstrated that an application of DPT on MN had the effects similar to those of dopamine application, as established earlier, causing the enhancement of MN resistance to fatigue stimulation, accompanied by an increase of the dimensions of the actin containing desmosome-like afferent admembranous synaptic contacts, and formation of the cytoplasmic bundles of actin stress-fibers. Similarity of morpho-functional changes of MN, induced by DPT, an artificial chemical substance, which has no receptors on the neuronal membrane, and by natural neurotransmitter dopamine, allows us to suggest possible trophic stabilizing and polymerizing effects of both substances on cytoskeletal actin due to their direct penetration into postsynaptic neuron.

[Cytotoxicity of amyloid fibrils of X-protein].

It is known that amyloid oligomers, protofibrils, and fibrils induce cell death, and antibiotic tetracycline inhibits the fibrillization of beta amyloid peptides and other amyloidogenic proteins and disassembles their pre-formed fibrils. Earlier we have demonstrated that sarcomeric cytoskeletal proteins of the titin family (X-, C-, and H-proteins) are capable to form in vitro amyloid fibrils, and tetracycline effectively destroys these fibrils. Here we show that the viability of polymorphonuclear leukocytes in the presence of X-protein amyloids depends on the concentration of amyloid fibrils of X-protein and the time of incubation. In addition to the disaggregation of X-protein fibrils, tetracycline eliminated the cytotoxic effect of the protein. The antibiotic itself did not show a toxic effect, and the cell viability in its presence even increased. Our results evidence the potential of this approach for evaluating the effectiveness of drugs preventing or treating amyloidoses.