[Regulation of intracellular lipid metabolism by the preparation of omega-3 phospholipids from marine organisms in deficiency of essential fatty acids in rats].

It has been established that a deficit of essential fatty acids (EFA) in the animal organism induces specific modifications of composition of fatty acid (FA) of general phospholipids and plasmalogenic P1 in microsomal tissue membranes with various functions and affects the activity of phospholipase A2. It has been shown that arachidonic (AA), docosapentaenoic (DPA) and docosahexaenoic (DHA) acids in the composition of general phospholipids - phosphatidylcholine (PC), phosphatidylethanolamine (PEA) and plasmalogens PC and PEA react to EFA deficit in the organism. Quantitative redistribution of AA, DPA, DHA of FA in general phospholipids and plasmalogenic microsomal membranes depending on their functions was found under EFA deficit in the organism. Deficit of DHA and plasmalogenic phospholipids evidences that the status of cell plasmalogens affects the level of PUFA at EFA deficit in the organism. AA and DHA can be a selective target for plasmalogens. The drug of omega-3 phospholipids, considerable amount of DHA and eicosapentaenoic (EPA)FA being present in their structure, increases the amount of plasmalogens and decreases the amount of AA in the brain, heart and reproductive organs. It was also found that EFA deficit in the organism favours the increase of lisoPEA, lisoPC, free FA (FFA) connected with the increase of activity of endogenic phospholippase A2 and plasmalogen-selective phospholipase A2. The omega-3 phospholipase from marine organisms at EFA defecit decreases the amount of FFA, lisophospholipids and activity of phospholipase A2 in the microsomas of the studied tissues. The drug of omega-3 phospholipids normalizes the state and functions of the brain, liver, and heart tissues, reproductive organs against a background of EFA defecit and regulates the synthesis of biologocically active metabolites of AA in the organism.

[Distribution of fatty acids of plasmalogenic phospholipids in human erythrocytes and endometrium in hyperplasia of various type]. / Rozpodil zhyrnykh kyslot plazmalohennykh fosfolipidiv v erytrotsytakh krovi ta v endometriï liudyny pry hiperplaziï riznoho kharakteru.

It is shown that development of hyperplasia of endometrium (PPPE) is accompanied by the valid decrease of plasmenyl-phosphatidylethanolamine (PL PhEA) amount both in the blood erythrocytes and human endometrium and by the increase of diacylglycerophosphoethanolamin (DG PhEA). Data obtained evidence for the development of the state of plasmalogenic phospholipids insufficiency in the organism. When investigating the composition and quantity of fatty acids of plasmalogenic phospholipids of PhEA in the blood erythrocytes of patients it was established that the amount of arachidonic (C 20:4 omega-6) and docosahexaenoic (C 22:6 omega-3) reliably decreases more than by 50%. At the same time one can observe the reliable increase of palmitic (C 16:0) and stearic (C 18:0) fatty acids. Under these conditions the ratios omega-6/omega-3 of both arahidonic and docosahexaenoic (AA/DHA) PL PhEA in the composition of erythrocyte PhEA increase with hyperplasia development 4 and 2 times respectively, but in DPhG the above ratios decreased 1.5 times. The decrease of PUFA in erythrocytes evidences for the increase of their use for restoration of their level disturbances in the organism. As a result of analysis of fatty acids content in tissues at hyperplasia development, considerable decrease of AA and DHA was established in one group of examined patients compared with the control, as well as the increase of omega-6/omega-3 2.8 times, and AA/DHA 2 times. Reliable double increase of AA and eicosatrienoic acid and decrease of DHA and linolenic acid was observed in group 2 of the patient with adenomatosic hyperplasia. The ratio omega-6/omega-3 and AA/DHA decreased twice in the both groups as compared with the control, that evidenced for development of hyperplasia of different origin. Specific distribution of arachidonic acid in plasmalogenic phospholipids at adenomatosic hyperplasia is connected with decrease of plasmalogens content and increase of formation of prostaglandin E2.

[Specificity of endometrium phospholipids and their fatty acids changes in the development of hyperplasia]. / Spetsyfichnist' zmin vmistu fosfolipidiv i ïkh zhyrnykh kyslot v endometriï pry rozvytku hiperplaziï.

Quantitative changes in phospholipids (PL) and fatty acids content (FA) in patients with endometrium hyperplasia have been revealed. During observing endomentrium hyperplasia process we found out on the one hand reliable decrease in phosphatidylethanolamine (PEA), phosphatidylcholine (PC) content and on the other hand increase of its lysoforms. Changes in unsaturated fatty acids (PA n-6)--arachidonic (AA) content and docosahaenoic (DHA) acids content were also shown. The decrease of PC, PEA in patients with endometrium hyperplasia is interrelated with quantitative change of plasmalogenic phospholipids PL, which is too much sensitive to oxidative reactions and at the same time able to protect sells against oxidative stress. Considerable decrease in plasmenyl PEA content on the one side and plasmenyl PC content stability on the other was revealed in patients with endometrium hyperplasia. Above mentioned lipid metabolic derangements of the endometrium are specific with respect to different classes of phospholipids and their fatty acids composition.

[Effect of phospholipids containing omega-3 fatty acids on structural changes of microsomal lipids in cell membranes of functionally different cells]. / Vplyv fosfolipidiv, shcho mistiat' omeha-3 zhyrni kysloty, na strukturni zminy lipidiv mikrosom membran klityn, riznykh za funktsiieiu.

As a result of the experimental researches conducted it has been shown that administration of some normal animal marine phospholipids (PL) including in their structure omega-3 polyunsaturated fatty acids (PUFA) provides for quantitative changes of individual PL, fatty acids (FA) content and quantity in general and individual PL of liver, heart, brain and gonads microsomes. While estimating general microsomal PL fraction FA content under the action of PL omega-3 PUFA FA concentration change, unsaturation index (omega 6/omega 3) and relation of arachidonic acid to docosahexenic (AA/DHA) decrease have been identified. The decrease of AA/DHA relationship occurs due to AA and DHA quantitative changes. In the case of AA increase in some tissues there is observed the decrease of docosapentaenic acid and increase of DHA and eucosapentaenic (EPA) acidds. As a result of studying FA content in the individual PL composition it has been identified that certain PL classes characteristic for some tissues respond by changes of some certain FA. The relationship omega 6/omega 3 has been shown as decreasing in phosphatidilcholine (PC) all tissues microsomes (liver, gonads, heart, brain), in phosphatidilethanolamine (PEA) of liver and cardiac microsomes, in phosphatidilserine (PS) this relationship relationship decreases in the liver, brain and heart, for phosphatidilinositole (PI) the changes take place in liver, gonads, brain. Simultaneously, the decrease of AA/DHA relationship in the individual PL decrease of AA and increase of EPA and DHA depend on the tested tissues. The marine phospholipids might be supposed to render their effect on AA metabolism resulting in AA/DHA relationship in PEA and PS relationship displays itself as specific and depends on the tissues functions. The preference of PEA and PS use by certain tissues microsomes could be explained by their membrane protective capability.

[Effect of "marine" phospholipids omega-3 fatty acids on the composition of fatty acids of rat liver microsomal membrane under oxidative stress]. / Vplyv "mors'kykh" fosfolipidiv z omega-3 zhyrnymy kyslotami na sklad zhyrnykh kyslot mikrosomal'nykh membran pechinky shchuriv za okysliuval'noho stresu.

As a result of experiments conducted the marine phospholipids preparation enriched by omega-3 fatty acids was defined to modify fatty acids content due to changes of fatty acids level change in the neutral lipids and phospholipids fractions. As well it was identified, that at the oxidative stress induced by administration of CCl4 the growth of arachidonic and docozahexaenoic acids in the neutral lipids fractions was observed if compare with the norm. At the same time, the presented fatty acids in the phospholipids fractions remained unchanged. At oxidative stress the phospholipids fraction reacts to levels of arachidonic and docozahexaenoic acids just only as a result of administrating phospholipids with omega-3 fatty acids. The most attractive is the change of correlation C20:4/C22:6--increasing at administration of CCl4 and decreasing both at phospolipids and vitamin E injection. Thus, at the oxidative stress the first reacting ones are the fatty acids of neutral lipids microsomal membranes.

[Biological properties of the mechanism of "marine" phospholipids containing omega-3 fatty acids]. / Biolohichni vlastyvosti ta mekhanizm diï "mors'kykh" fosfolipidiv z omega-3 zhyrnymi kyslotamy.

The Laboratory mainly deals with the development of biologic technologies for producing physiologically active lipid-protein-nature compounds from marine organisms containing omega-3 fatty acids (that are membrane components) with further evaluation of their action both in the normal state and in some simulated pathologic states as well as with creation of new preparations for application in medicine and agriculture on their basis. As a result of the experiments performed, a technology for producing two biologic preparations, namely, surface active compounds (phospholipids) and a nucleopeptid-lipid complex, exhibiting a specific action, was developed. The phospholipid complex, being surface-active in composition, was characterized as a complex possessing some surfactant-type properties and displaying an antioxidant and a membrane-stabilizing effect. On the basis of the complex including marine phospholipids with omega-3 fatty acids, the preparation "Phylomek", which is the concentrate of essential marine phospholipids, and the preparation "Morephyl", which is a marine agent with surfactant-type effect, intended for animals and poultry, were created. The ingredients of the nucleopeptid-lipid complex were identified, and its effect on the increase of testosterone levels in the blood of old and sick animals was determined. A stimulating agent of genital hormones secretion was recommended for application in the geriatrics in the case of genital function disturbance and presenilation. A fraction similar in chemical content and specific activity, found in the velvet antlers, was used as the base of the biologic preparation "Pantheron". The natural complexes of marine phospholipids were shown to be able to change the composition of lipids of membranes, but the intensity of these changes differed in cells differing in function. In the study of the biochemical mechanism of correction of disturbances in the cell membranes under an oxidative stress, the interrelation between the composition of lipids of membranes, their oxidation, and the content of natural antioxidants was determined. The reparative effect of marine phospholipids on the cell membranes under progressing pathology, caused by the restoration of the composition of phospholipids, by increases in the activities of antioxidant enzymes (SOD, catalase), and by a decrease of the accumulation of LPO products, was established. Under interaction of marine phospholipids and alpha-tocopherol, synergism was noticed, the antioxidant potential of the investigated substances and their membrane-stabilizing effect increased. The phospholipids with various residues of PUFA in the molecule were found to affect the inhibition and oxidation processes, as well as the modelling of lipid membranes. This is especially true for arachidonic and docosahexaenoic acids, the ratio of which changes under the oxidative stress. At administration of phospholipids omega-3, their ratio decreases due a decrease in the level of unetherificated PUFA. The main changes of the PUFA were found to occur in phosphatidylethanolamine isolated from microsomes. The particular role of phosphatidylethanolamine and arachidonic acid in the reparation of membranes under the action of phospholipids PUFA omega-3 and alpha-tocopherol was noted.

[Role of phospholipids in functionally different cell membranes under conditions of antioxidant system disturbance]. / Rol' fosfolipidiv u membranakh funktsional'no riznykh klityn za porushennia antyoksidantovoï systemy.

Vitamin E deficiency in organism leads to microsome membrane phospholipid composition changes, oxidation, superoxide dismutase (SOD) activity decrease, accompanied with the disorders of lung and liver tissues functional state which is connected with regulation of membrane penetration. The negative effect of E-hypovitaminosis more affects the liver than lung, which is caused by their different functional purposes, peculiarities of metabolic processes as well as by the content of biomembrane structural components. Vitamin E protects phospholipids from oxidation, regulates AOS condition, SOD-activity and promotes in such way the tissue functional disturbances correction of the animals tested. New preparation "Calmophyl" is a phospholipid complex from marine organisms with surfactant-kind effect, which displays antioxidative properties and decreases toxic affect of CCl4, corrects functional tissue disorders, regenerates normal activity of enzymes. Preparation of natural phospholipids has been shown to be capable to stabilize cell membranes probably by means of replacing the membrane phospholipid composition and decreasing disturbances in the antioxidative system.

[Superoxide dismutase activity in microsomes of functionally different rat tissues in vitamin E deficiency]. / Aktivnost' superoksiddismutazy v mikrosomakh funktsional'no razlichnykh tkanei krys pri nedostatochnosti vitamina E.

Comparative investigation of microsome superoxide dismutase activity in functionally different rat tissues such as lung and liver has been carried out. Modification of superoxide dismutase fraction isolated from lung and liver microsomes and SOD-activity evaluation by means of superoxide radical-dependent pyrogallol autooxidation method have been described in detail. SOD-activity has been shown to be connected with the degree of lipid peroxidative oxidation (LPO) and total condition of tissue antioxidative system in case of different E-vitamin security in various functional types of cells (alveolocytes and hepatocytes).

[Laboratory of technology of biopreparations]. / Laboratoriia tekhnolohiï biopreparativ.

The main scientific direction of the Laboratory is the development of new biochemical technologies for obtaining various biopreparations based on animal and plant raw materials, especially on that of the sea organisms. Fundamental investigations of the preparations of animal and microorganism hydrolysis have enabled the researchers to develop technologies for obtaining Str. griseus and Ceph. acremonium proteolytic complexes and to study their properties for the latter could be used as reagents in chemistry of proteins. Immobilized polyenzyme systems of proteases with silicagel and activated carbon fibre material as a matrix were created on the basis of investigations of immobilized enzymes. The advantages of immobilized biocatalysts possessing highest stability and a possibility of repeated application are described. Biotechnological isolation of bioactive preparations (BAP) of lipid-protein nature that are the structure components of cells membranes is the key problem at present. Biochemical principles of BAP metabolisms regulation in cell membranes and the role of the obtained biopreparations in correction of pathological conditions are also studied. These investigations promoted development of technologies for two new biopreparations from sea organisms (Calmar's gonades) for medicine. The first one is a set of surface active phospholipids and the second one is a set of nucleopeptides affecting the secretion of sex hormones. It has been found that surface active preparations show an antioxidative and membrane-stabilizing properties as well. It has also been shown that the preparation corrects the effect of pathology conditions in case of experimental hepatitis-induced by CCl4. The preparation influences the antioxidative system and thus the rate of lipid peroxidation.(ABSTRACT TRUNCATED AT 250 WORDS)

[Antioxidant effect of a phospholipid complex isolated from marine organisms]. / Antioksidantnoe deistvie fosfolipidnogo kompleksa, vydelennogo iz morskikh organizmov.

Antioxidative effect of phospholipid complex from the sea organisms (preparation "Kalmofil") is observed comparatively with other phospholipid fractions of animal and plant origin. Most antioxidative action of the phospholipid complex from Mollusca (preparation "Kalmofil") on the nonenzymatic LPO reactions in vitro is established. Analogous fractions from different functional animal tissues have not such influence. Application of "Kalmofil" in vivo under experimental rat hepatitis provoked by CCl4 has a corrective effect on both nonenzymatic and Fe(2+)- and NADP.H-depending LPO reactions. This preparation decreases toxic influence of CCl4 and improves antioxidative system of rat hepatocytes.

[Hypolipidemic action of liposome-incorporated beta-sitosterol on rats]. / Gipolipidemicheskoe deistvie zakliuchennogo v liposomy beta-sitosterina u krys.

Daily administration to rats with experimental hypercholesterolemia of beta-sitosterol (5 mg/kg) contained by lecithin liposomes provoked a 25% reduction in blood plasma cholesterol in 3 weeks, whereas crystalline beta-sitosterol (50 mg/kg) produced an analogous effect in 6 weeks. The drug under study decreased the level of atherogenic beta-lipoproteins in blood plasma and intestinal mucosa, raised the content of phospholipids in blood plasma and normalized the cholesterol/phospholipid ratio. The drug high hypolipidemic activity was supported in experiments in vitro with inverted sacs of the small intestine. The drug was found to be long-acting.

[Effect of beta-sitosterol incorporated into liposomes on several indicators of lipid metabolism in experimental hypercholesteremia in rats]. / Vliianie beta-sitosterina zakliuchennogo v liposomy, na nekotornye pokazateli lipidnogo obmena pri éksperimental'noi giperkholesterinemii u krys.

Alterations in lipid content of microvilli membranes, affecting their physical structure, as well as in lipoproteins of rat small intestinal mucosa and of blood plasma were studied in experimental hypercholesterolemia after treatment with crystalline and liposomal preparations of beta-sitosterol. The liposomal preparation of beta-sitosterol exhibited distinctly higher hypocholesterolemic activity as compared with the crystalline sterol. Possible role of structural and functional alterations in enterocyte membranes and in intestinal lipoproteins during absorption of cholesterol as well as inhibition of the processes by means of beta-sitosterol are discussed.

[Hypocholesterolemic effect of phytoecdysones during experimental hypercholesterolemia in rats]. / Gipokholesterinemicheskoe deistvie fitoékzidonov pri éksperimental'noi giperkholesterinemii u krys.

The most active hypocholesterolemic preparation, affecting the absorption of 3H1-cholesterol in small intestine of rats with experimental hypercholesterolemia, proved to be I-hydroxyecdisterone; the activity was gradually decreased in the following series of preparations: ecdisterone, steroid sapogenine allyogenone, ciasterone and ecdisterone 25 acetate. Thus, the hypocholesterolemic activity of the preparations was increased with an increase in the content of hydroxy groups in the molecules. After daily administration of ecdisterone at a dose of 2.5 mg/kg into the animals with hypercholesterolemia within 3, 6 and 8 weeks, content of cholesterol in blood plasma was decreased by 7.0%, 16.9% and 29%, respectively; this phenomenon was accompanied by a decrease of the cholesterol content in erythrocyte membranes as well as in microfilaments of erythrocyte border by 26% and 34%, respectively. Amount of phospholipids and the ratio cholesterol/phospholipids were also normalized in the membranes studied. The data obtained suggest that competitive relationship occurs between cholesterol and the hypocholesterolemic drugs during binding with the membrane sites depending on their concentrations in blood plasma, intestine lumen and on the content in the membranes.

[Lipid composition of different preparations of brain Na+, K+-ATPase]. / Lipidnyi sostav razlichnykh preparatov Na+, K+ -ATP-azy golovnogo mozga.

The content and composition of phospholipids is determined in beef microsomal and synaptosomal fractions and also in these fractions preparations solubilized with triton X-100 (0.1%) and digitonin (0.2%). It is shown that the microsomal fraction is richer in phospholipids. The solubilized fragments of microsomes have less or the same amount of phospholipids per protein unit than the initial fraction of microsomes, and the solubilized fragments of synaptosomes contain a higher quantity of phospholipids than the initial fraction. The content of phospholipids in "the riton" fragments of synaptosomes is higher than in "those" of microsomes. Contrary to digitonin which solubilizes the active Na+, K+-ATPase complex of microsomes and synaptosomes, triton X-100 solubilizes the active enzyme of microsomes only. A higher total content of phospholipids in "the triton" extracts of synaptosomes does not probably correlate with the presence of Na+, K+-ATPase activity in them. But these extracts are found to contain less phosphatidylserine whose addition recovers Mg2+, Na+, K+-ATPase activity in them. The effect of phosphatidylserine is not strictly specific for "the triton" extracts of synaptosomes, this lipid activates to a definite extent the extracts of microsomes as well. It is shown that at the first stages of bull brain Na+, K+-ATPase purification the total content of phospholipids and cholesterol in the preparations increases but the composition of phospholipids remains unchanged.

[Lipid composition and activity of certain enzymes in membranes of intestinal epithelium microvilli in rats with experimental hypercholesterinemia]. / Lipidnyi sostav i aktivnost' nekotorykh fermentov membram mikrovorsinok kishechnogo épiteliia krys pri éksperimental'noi giperkholesterinemii.

It is established that in membranes of small intestine epithelial cells microvilli in rats, which were administered cholesterol for a long time, the content of this sterol and its esters as well as of phospholipids is different. Differences in the content of lipids are a reason of changes in the sucrose, alkaline phosphatase and leucilamino-peptidase activities. These changes are accompanied by an increase on the molar cholesterol esters-free cholesterol ratio in blood plasma and cholesterol-phospholipids ratio in the microvilli membranes. Beta-sitosterol administered to experimental animals incorporates into the intestine plasma membranes, blocks up the transfer sites, inhibiting cholesterol absorption, which evidently decreases the sterol content in blood serum.

[Role of sterol photoconversion products in the regulation of cholesterol biosynthesis in rat skin]. / Rol' produktov fotoprevrashcheniia sterinov v reguliatsii biosinteza kholesterina v kozhe krys.

In order to clarify the effect of products of photochemical conversion of sterols on cholesterol biosynthesis, rat skin samples were incubated with 2-(14)C-acetate in the presence of the antirachitic agent Dk and 7beta-hydroxycholesterol. The synthesis of sterols from acetate was activated in the presence of Dk. A correlation between the activation of sterol synthesis and the concentration of the antirachitic agent was found. An addition of 7beta-hydroxycholesterol to the incubation medium inhibited acetate incorporation into the sterols. The level of synthesis inhibition increased with an elevation of the 7beta-hydroxysterol concentration in the incubation medium. This indicates that both products of sterol photoconversion can be involved in the control of cholesterol biosynthesis.

[Cholesterol synthesis in the skin of rats with rickets]. / Syntez kholesterynu v shkiri shchuriv za rakhitu

The rachitogenic diet causes an inhibition of cholesterol synthesis in rat skin. Intensity of 2-14S-acetate incorporation into cholesterol precursors (matastenol, lathosterol, 7-dehydrocholesterol) and cholesterol drops considerably. UV-irradiation of animals suffering from rachitis intensifies cholesterol synthesis in skin. Simulteneously an increase is detected in the metabolic fund of cholesterol synthesis precursors, including 7-dehydrocholesterol, which is necessary to form antirachitic compounds in the organism.

[Effect of ultraviolet irradiation of different spectral composition on cholesterol synthesis in rat skin]. / Vplyv ul'trafioletovoho oprominennia riznoho spektral'noho skladu na syntez kholesterynu v shkiri shchuriv

Irradiation of rats with of UV-radiation sources epsilonYB-15, (see article) causes a decrease in the intensity of 2-14C-acetate incorporation into cholesterol of skin and small intestine and simultaneous accumulation of radioactivity in its predecessors, namely in lathosterol and 7-dehydrocholesterol. Thus, it is shown that UV-irradiation of animals inhibits the cholesterol synthesis in skin and tissue of the small intestine. Intensity of cholesterol synthesis inhibition with UV-irradiation of animals depends on the spectral, composition of the acting radiation.

[In vitro biosynthesis of cholesterol and its precursors in the osseous tissue of rats]. / Issledovanie biosinteza kholesterina i ego predshestvennikov v kostnoi tkani krys in vitro

By thin-layer chromatography compounds of sterol origin were isolated from the nonsaponified fraction of bone tissue of rats. The occurrence of squalene, lanosterol, cholesterol and incompletely identified Cdelta2427-sterol in the organic matrix of the bone was demonstrated by means of UV- and IR-spectrometry. Bone tissue incubation with 1,2-C14-aceticacid yielded C14-labeled lanosterol, C28, 29-sterols, cholesterol and squalene. The rate of cholesterol synthesis in the bone tissue was lower than that in the liver and intestine of rats.