[Cardioprotective effect of N-stearoylethanolamine under the anaphylactic shock in guinea pigs].

It was shown for the first time that endogenous cannabimimetic compound N-stearoylethanolamine (NSE) administered at the dose 65 mg/kg of body weight during two weeks before anaphylaxis had cardioprotective properties on the model of anaphylactic shock in the guinea pigs. NSE reliably normalized the content of histamine and nitric oxide, decreased the activity both inducible and constitutive NO-synthases, prevented accumulation of thiobarbituric acid reactive substances (TBARS) and normalized the activity of catalase, superoxide dismutase and glutathione peroxidase in cardiomyocytes of animals under anaphylaxis. About of 70% of the NSE-treated animals survived under the anaphylactic shock. The present work indicates that NSE may influence the allergic reaction of immediate type and can possibly be used for design of a new type of antiallergic preparations and cardioprotective preparations as well.

[Changes of nitric oxide level at different stages of primary open-angle glaucoma]. / Izmenenie urovnia oksida azota na raznykh stadiiakh pervichnoi otkrytougol'noi glaukomy.

The contents of NO stable metabolites in tears, and blood serum of the patients with different stages of open-angle glaucoma have been studied. The increasing of NO stable metabolite contents was found in tears, aqueous humor and blood serum during glaucoma progression. The depletion of glaucoma pathogenesis oxidative link was found at the terminal stage of the disease. The results of our investigation are proposed to be used for glaucoma diagnostic.

[Relationship between polyreactive properties of immunoglobulins and level of lipids in them]. / Vzaimosviaz' mezhdu poliraktivnymi svoistavmi immunoglobulinov i soderzhaniem v nikh lipidov.

Presented paper deals with the relationship between immunoglobulin polyreactive properties and its lipid composition. Serum blood immunoglobulin fraction of an intact rabbit as an experimental model was used. Immunoglobulins (Ig) obtained by this way were transformed into polyreactive immunoglobulins (PRIg) by treatment with chaotropic agent KSCN or reactive oxygen species (ROS) with usage of Fe2+, EDTA and ascorbic acid. It was demonstrated that native Ig were able to bind with immobilized antigen (ovalbumin) and this ability dramatically increased after transformation of Ig into PRIg. The high immunoreactivity of PRIg was associated with marked fall (by 2-3 fold) of total phospholipids as well as individual ones--sphingomyelin and phosphatidylcholine. The main fatty acids of the Ig and PRIg phospholipid fractions in the sequence to decreasing decrease were stearic, palmitic, oleic and linoleic acids. The treatment of Ig by chaotropic agent and ROS led to decrease of stearic acid and enhancement of oleic, linoleic and arachidonic acids. The level of free cholesterol of Ig did not differ from that of PRIg. At the same time the content of cholesterol esters of PRIg was substantially diminished if compare with Ig. The main fatty acids of the Ig and PRIg cholesterol ester fraction in the sequence to decreasing were arachidonic, stearic, oleic, linoleic and palmitoleic acids. Transformation of Ig into PRIg was accompanied by enhancement of stearic acid level and loss of docosapentaenoic, arachidonic and palmitoleic acids. The results presented here support the idea about non peroxidative manner of the phospholipid and cholesterol ester extrusion from Ig molecule under its transformation into PRIg. Rather the last process could be explained by the term of concurrent physico-chemical interaction of Ig molecule with chaotropic agent or ROS leading to fall of lipid content. The presented data for the first time provide us an opportunity to conclude that transformation of Ig into PRIG is associated with the marked loss of essential phospholipids and cholesterol esters by the Ig molecule. The probable implication of this process in development of immune imbalance under certain diseases associated with oxidative stress have been discussed.

[Characteristics of the effect of implantation of the human ApoA1 gene on the condition of hepatocytes in rats of varying age]. / Osobennosti vliianiia implantatsii gena ApoA1 cheloveka na sostoianie gepatotsitov krys raznogo vozrasta.

The results obtained show the essential changes in functional state of hepatocyte's plasmatic membrane due to the implantation of human ApoA1 gene to the rat liver. The changes in phospholipid composition, hyperpolarization, increase in activity of membrane bound enzymes, cytochrome P-450 and biosynthesis of liver total proteins have been found. The essential changes characterizing cell effect were more marked in the adult rats, and membrane effect in the old ones.

[Effect of N-palmitoylethanolamine on energy-dependent transport of Ca2+ in vesicles of myometrium sarcolemma and their phospholipid composition]. / Vliianie n-pal'mitoilétanolamina na énergozavisimyi transport Ca2+ v vezikulakh sarkolemmy miometriia i ikh fosfolipidnyi sostav.

N-palmitoylethanolamine (NPE) was studied for their effect on calcium pump of pig myometrium sarcolemma. NPE in concentration of 10 microM, stimulated by 28-46% Mg2+, ATP-dependent accumulation of Ca2+ in vesicles of plasmatic membrane of uterus myocytes taking absolutely no effect on passive release of this cation from them. NPE modified phospholipid composition of sarcolemma, causing the increase of percentage content of phosphatidylinositol (by 20.2%) and lysophosphatidylcholine (2.7 times). While NPE effects transport Ca2+, Mg(2+)-ATPase solubilized from plasmatic membrane and purified due to the method of affinity chromatography on calmodulin-sepharose 4B, no activating effect of NPE on the calcium pump was observed. And what is more, a weakly expressed tendency to inhibition (by 14-15%, respectively) of the rate of Ca2+, Mg(2+)-dependent enzymic hydrolysis of ATP has been revealed. It is supposed that the effect of NPE on active transmembrane transport of Ca2+ is an important link in the general mechanism of contraction-relax of the myometrium and is, apparently, connected with its modifying effect on the lipid composition of the sarcolemma.

[Effect of n-palmitoylethanolamine on energy-dependent transport of Ca2+ in intracellular structures of myometrium and their phospholipid composition]. / Vliianie n-pal'mitoilétanolamina na énergozavisimyi transport Ca2+ vo vnutrikletochnykh strukturakh miometriia i ikh fosfolipidnyi sostav.

N-palmitoylethanolamine (NPE) was studied for its effect on the systems of energy-dependent transport of Ca2+ in the intracell structures (mitochondria, endoplasmic reticulum) of permeabilized cells of smooth muscles of the rat uterus as well as on the lipid composition of myocytes. NPE in concentration of 10(-5) M partially (by 30-50%) inhibited energy-dependent accumulation of Ca2+ in endoplasmic reticulum and mitochondria of myometrium cells permeabilized by means of treatment of myocytes suspension by digitonin (0.1 mg/ml). NPE modifies the lipid composition of permeabilized myocytes when determining the increase of the amount of inorganic phosphorus of total phospholipids by 57.3% at the expense of considerable accumulation of phosphatidylcholine, phosphatidylethanolamine and sphingomyelin. It is supposed that the effect of NPE on the systems of energy-dependent transport of Ca2+ is achieved through the modification of phospholipid composition of a cell, while modulating effect of NPE on the active transmembrane transfer of Ca2+ in the intracellular structure can be an important link in the general mechanism of the effect of this compound on Ca2+ metabolism in myometrium and on Ca(2+)-depended control of the contracting function of the uterus.

[Effect of N-palmitoyl- and N-stearoylethanolamines on lipid peroxidation in mouse tissues in metastatic Lewis carcinoma]. / Vliianie N-pal'mitoil- i N-stearoilétanolaminov na perekisnoe okislenie lipidov v tkaniiakh mysei pri metastazirovanii kartsinomy L'iuis.

The ability of two-cell minor lipid components: N-palmitoyl- and N-stearoylethanolamines to influence the Lewis lung carcinoma metastases has been studied. It is shown that these compounds decrease the quantity and the volume of metastases. Acyl derivatives of ethanolamine inhibit simultaneously the process of lipid peroxidation. Mice C57 B1/6 were intraperitonealy injected labeled 3H-N-stearoylethanolamine. Localization of this radioactive label in some organs of the animals was studied.

[The characteristics of lectin binding to the surface of differentiated neuroblastoma C 1300 cells]. / Osobennosti sviazyvaniia nekotorykh lektinov s poverkhnost'iu differentsirovannykh kletok neiroblastomy C 1300.

Peculiarities of the binding of lectins of castor bean, cochlea, soya bean, lime bean and wheat germs to the surface of differential neuroblastoma C 1300 N 18 cells have been studied using the method electron cytochemistry. It is found that the quantity of the bound lectin conjugates with colloidal gold on the surface of differentiated cells varied considerably from that on the surface of nondifferentiated cells.

[Morphological differentiation and change in the lipid composition of neuroblastoma C1300 cells treated with gangliosides]. / Morfologicheskaia differentsirovka i izmenenie sostava lipidov kletok neiroblastomy C1300 pod vliianiem gangliozidov.

The effect of exogenous gangliosides on the morphological differentiation of neuroblastoma 1300 N18 cells was studied Simultaneously the content of gangliosides and lipid composition of the cells was investigated. Gangliosides were shown to increase the quantity of cells with long neurites. This effect depended on the dose of gangliosides. The addition of 50 and 100 micrograms of gangliosides per 4 ml of serum-free culture medium increased the quantity of cells with neurites by 38 and 63.4%, respectively. The level of morphological differentiation in cells cultivated with gangliosides was higher than in cells incubated with 5'-bromodeoxyuridine. Noticeable quantities of lysophosphatidylcholine (absent in the control) appeared in ganglioside-treated cells and the level of cholesterol increased. The amount of other lipid compounds in cells differentiated in the presence of gangliosides was similar, but not identical to the quantity of lipid compounds in cells differentiated by 5'-bromodeoxyuridine and by the serum-free medium.

[Detection of N-acylethanolamine phospholipids in neuroblastoma C1300 cells]. / Obnaruzhenie N-atsilétanolaminovykh fosfolipidov v kletkakh neiroblastomy C1300.

Phospholipids with high chromatographic mobility are detected in cells of murine neuroblastoma C 1300. One of them is identified (proceeding from data of successive chemical degradation) as N-acylphosphatidylethanolamine. It may be a precursor of physiologically active N-acylethanolamines. N-palmitoyl and N-stearoylethanolamines are shown to inhibit the transport of ions through the veratrine-activated fast sodium channels as well as remove the effect of lysophospholipids accumulation in cells under the effect of veratrine.

[Identification and properties of insulin receptors in neuroblastoma C 1300 N 18 cells in various functional states]. / Identifikatsiia i svoistva insulinovykh retseptorov v kletkakh neiroblastomy C 1300 N 18 pri razlichnykh funktsional'nykh sostoianiiakh.

The presence of insulin receptors in neuroblastoma C 1300 N18 cells was shown by the method of the [125I]insulin binding with cells as well as by the electron-cytochemical methods based on the analysis of the binding of colloid gold-labelled insulin and of agglutinin of wheat embryos with the surface of plasma membrane. It is established that the number and distribution of insulin receptors depend on the functional state of cells. Expression of receptors on the membrane increases after 5'-deoxyuridine-induced differentiation of cells. Due to changes in the lipid composition of cells caused by the incubation with lecithin-cholesterol liposomes (an increase in the content of cholesterol and its esters, as well as of unsaturated fatty acids and appearance of lysolecithin) the quantity of insulin receptors decreases and cell membranes are damaged. The lowering insulin regulation is not observed for insulin receptors of the neuroblastoma C 1300 N18 cells.

[Localization and functional role of gangliosides from nerve tissue of vertebrates]. / Lokalizatsiia i funktsional'naia rol' gangliozidov nervnoi tkani pozvonochnykh.

Gangliosides in the animal organism are typical components of plasma membranes of nerve cells in which the concentration of polysialogangliosides is especially high. The high concentration of tri- and tetrasialoganyliosides and ganglioside GD1b is peculiar to primary cultures of nerve cells, whereas these gangliosides are practically not present in the culture of the transformed nerve cells which have lost their ability to sinaptogenesis, they are not found in cultures of oligodendro- and astroglia as well. The addition of exogenic gangliosides to nerve cells cultures stimulates the formation of processes in these cells and promotes their survivability. The neuritogenic and neuronotrophic effect of gangliosides, their participation in the processes of neurons' regeneration are shown in the in vivo experiments. Gangliosides are carriers of antigenic determinants typical of the cellular surface of neurons (or other cells) as well as of cells of different malignant tumours; typical carcinoembryonal antigenes are revealed among them. Such functions of gangliosides as participation in the processes of intercellular interaction, adhesion, pneuritogenic effect, possible participation to memorize one or another habit are, probably, interrelated and mutually conditioned. Recently the data, being in controversy with the notion that gangliosides are components of receptors of hormones and mediators, are obtained. Evidently, it ought to speak about the modulation of cell response by them on the action of these effectors.

[Transmembrane lipid transport between lecithin liposomes and neuroblastoma C1300 N18 cells]. / Transmembrannyi transport lipidov mezhdu letsitinovymi liposomami i kletkami neiroblastomy C1300 N18.

The method of double isotopic labels was used to study dynamics of lipid metabolism between neuroblastoma C 1300 N 18 A 1 cells and lecithin liposomes which contained 4.5-5 mumol of lecithin in 1 ml of the suspension. The cell lipids were labelled by radioactive carbon and cultivated on the medium with [1-14C] sodium acetate, phosphatidylcholine of liposomes was labelled by tritium. It is shown that 15-30 min long incubation with liposomes causes a sharp decrease of the cholesterol esters amount with a simultaneous fall of the free cholesterol level. The total content of phospholipids in this case remains unchanged though there occurs the noticeable exchange of labelled phospholipids between cells and liposomes. The cholesterol content in the plasma membranes of cells lowers sharply. The neuroblastoma cells are able to compensate arising changes in the cholesterol level for 45-60 min after which they progressively die. 90 min later only an insignificant part of the population (about 10% of cells) is retained.

[Inorganic pyrophosphatase activity of the mouse spleen in the immune response and after treatment with bis-phosphonates]. / Aktivnost' neorganicheskoi pirofosfatazy selezenki myshei pri immunnom otvete i pod vliianiem bisfosfonatov.

The inorganic pyrophosphatase activity was determined in different tissues of mice. The immunization of mice by sheep erythrocytes increased the inorganic pyrophosphatase activity of the spleen. The in vivo administration of bisphosphonates (40 mg per 1 g of mass), which are structural analogs of inorganic pyrophosphate (methylene bisphosphonic acid--MBPA, hydroxyethylidene bisphosphonic acid--HEBPA and aminomethylene bisphosphonic acid--AMBPA), inhibited the inorganic pyrophosphatase activity only by MBPA in the thymus and spleen but not in liver. The addition of MBPA, HEBPA as well as of phosphonoacetic acid, imidobisphosphate, bis(phosphonomethyl)-phosphonic acid, MBPA and phosphoric acid monoanhydride to cytosol from the mouse spleen led to the competitive (relative to the [Mg (PPi)2-] complex) inhibition of the inorganic pyrophosphatase activity. AMBPA didn't possess the analogous effect.

[Modification of the lipid composition of neuroblastoma C1300 cells using liposomes]. / Modifikatsiia lipidnogo sostava kletok neiroblastomy C1300 s pomoshch'iu liposom.

An essential increase in the total cholesterol content with a significant growth in the level of its esters was observed in neuroblastoma C1300 cells differentiated by means of 5'-bromodesoxyuridine and incubated for 1 h with lecithin-cholesterol liposomes (1:1 mol/mol). These cells also displayed an increased amount of polyunsaturated fatty acids in the composition of lecithin molecules and the appearance of lysolicethin as well. Incubation of the cells with lecithin liposomes was accompanied by a decrease in the content of cholincholesterol in the cells. In this case the level of cholesterol esters decreased to a greater extent than the level of free cholesterol. In lecithin molecules of these cells there occurred a relative increase of saturated fatty acids. The cells can retain viability and compensate changes caused by cholesterol excess during their incubation with lecithin-cholesterol liposome for 60-90 min. A longer incubation with liposomes results in a sharp drop of cell viability.

[Cultured neuroblastoma cells in biochemical studies]. / Kul'tura kletok neiroblastomy v biokhimicheskikh issledovaniiakh.

Neuroblastoma is a spontaneous tumour of animals and people. Tumour strains maintained as transplanted tumours and constant cell lines in suspension and monolayer cultures in vitro are obtained. The neuroblastoma cells are able to spontaneous and induced differentiation. The differentiated neuroblastoma cells have many specific properties of nature neurons and are a rather valuable object for solution of many problems in neurochemistry and general cell biology. It is neuroblastoma that has been used to show first the reversibility of neoplastic transformation and to carry out numerous studies to elucidate the mechanisms of cell differentiation. The neuroblastoma cell cultures are a convenient object for studying the ion channels and specifying the role of membrane lipids in their work.

[Relation between the functional characteristics of fast sodium channels and lipid composition of neuroblastoma C1300 cells]. / Zavisimost' funktsional'nykh kharakteristik bystrykh natrievykh kanalov ot lipidnogo sostava kletok neiroblastomy C1300.

The paper deals with functional characteristics of fast sodium channels modified by liposomes of differentiated neuroblastoma cells. Cells with a 60% increased total cholesterol content were used. The cells were found to contain lysolecithin. In lecithin the amount of polyunsaturated fatty acids was enhanced. It is established that veratrin added to the culture medium of cholesterol-enriched cells increased the rate of 86Rb release to a greater extent than in the control cells. When studying the temperature dependence of veratrin-activated transport of Rb+ in the intact and lecithin-cholesterol-lyposomes-modified cells it was shown that the veratrin-activated Rb+ release is more sensitive to temperature fluctuations than nonactivated one. In this case it is higher in lecithin-cholesterol-lyposomes-modified cells than in intact ones.

[Various physico-chemical properties of inorganic pyrophosphatase from the mouse spleen]. / Nekotorye fiziko-khimicheskie svoistva neorganicheskoi pirofosfatazy selezenki myshi.

Manganese, calcium and mercury ions, as well as p-chloromercury benzoate and dithiothreitol are studied for their effect on the activity of inorganic pyrophosphatase (EC 3.6.1.1) of mice spleen. It is shown that Ca2+ and Mn2+ are inhibitors of this enzyme, but Mn2+ in low concentrations may replace Mg2+ in the pyrophosphatase reaction. Hg2+ and p-chloromercury benzoate inhibit the pyrophosphatase activity essentially but not completely. Mice spleen pyrophosphatase is very labile: its preincubation without the substrate for 30 min at 37 degrees C leads to a complete loss of the activity. Neither glycerol, nor glutathione and cysteine but magnesium ions, dithiothreitol and 2-mercaptoethanol protect the enzyme from inactivation. The enzyme is purified by the sulphate ammonium salting-out, gel filtration on Sephadex G-100 as well as by isoelectrofocusing in 5% PAAG. Then pyrophosphatase is eluted from gel and subjected to electrophoresis in the plane layer of the linear gradient of 5-15% PAAG with SDS or 5-25% PAAG without denaturing conditions. One zone corresponding to the molecular mass of 70 kDalton is obtained. It is splitted into two zones in electrophoresis with SDS and 2-mercaptoethanol.

[Changes in the lipid composition of murine neuroblastoma C 1300 cells during the process of differentiation]. / Izmenenie lipidnogo sostava kletok neiroblastomy myshi C 1300 v protsesse differentsirovki.

The paper deals with the lipid composition of nondifferentiated cells at the logarithmic and stationary growth phases as well as with differentiated cells of C 1300 neuroblastoma adopted to the Eagle medium. It is shown that phospholipids and cholesterol dominate at the studied growth phases among lipids. Their amount in the differentiation process is thrice as high calculating per cells. The quantity of individual phospholipids changes in different manner during differentiation. The phosphatidylinosite level is higher than of other substances.

[Transport of Rb+ via activated sodium channels of clone N 18 phi 1 neuroblastoma cells]. / Transport Rb+ cherez aktivirovannye natrievye kanaly kletok neiroblastomy klona N 18 fi 1.

A study was made of the Rb+ transport via activated sodium channels of clone N 18 phi 1 neuroblastoma cells cultured in the Eagle medium with 10% bovine serum. The time of population doubling was about 10 h. The cell differentiation was induced by adding bromdeoxyuridine in a concentration of 1-4 10(-5) M. The cells contained 172 +/- 12 and 340 +/- 35 micrograms of protein per 10(6) cells at the logarithmic growth phase and in differentiated state, respectively. It is shown that veratrin produced a 1.3-fold increase in the rate of 86Rb+ removal from undifferentiated cells and 2.5-fold increase in that from differentiated cells. Tetrodotoxin removed completely the effect of veratrin. A conclusion is made on the presence of a new clone of fast sodium channels in cell membranes.