[The evaluation of unsaturation of blood lipids using methods of physical chemistry and clinical biochemistry. The insulin regulation of metabolism of fatty acids, number of double binds and cell absorption of glucose].

It is supposed that the main cause of insulin synthesis at late stages of phylogenesis became discrepancy between increase in vivo need in energy and physical chemical parameters of palmitic saturated fatty acid; its transportation to cells in composition of lipoproteins in optimal quantity (more than 15% of all fatty acids) became in vivo unfeasible. The biological role of insulin consists in supporting of insulin-dependent cells (skeletal miocytes in the first place) with substrates for gaining energy. The hormone transforms all palmitic saturated fatty acid endogenously synthesized from glucose into specific for animal cells rn-9 C18:1 oleic mono unsaturated fatty acid. The endogenous mono unsaturated fatty acid is oxidized by mitohondria with the highest constant of reaction velocity gaining for cells optimal quantity of biotransforming energy in the form of ATP. The insulin expresses in hepatocytes synthesis of oleic triglycerides and formation of oleic lipoproteins of very low density that only insulin-dependent cells absorb using apoE/B-100-endocytosis. The insulin expresses synthesis of Palmitoyl-KoA-elongase, stearyl-KoA-desaturase and glucose transporters 4, activates glucose absorption by cells with the purpose of synthesis endogenous oleic saturated fatty acid. The insulin substitutes in vivo ineffective palmitic alternative of metabolism of fatty acids for potentially more effective oleic metabolism of fatty acids. The insulin increases unsaturation of fatty acids and number of double binds in them. This can be established by direct titration of double binds by ozone on the basis of quantitative detection of fatty acids using technique of gas chromatography and calculating ratio C16:1/C16:0, C18:1/C18:0 and C18:1/C16:0. The diabetes mellitus is a disorder of metabolism of mono unsaturated fatty acid in the first place and only in the second place pathology of glucose absorption by cells.

[The oleic triglycerides of palm oil and palmitic triglycerides of creamy fat. The reaction of palmitoylation, potassium and magnesium palmitate, absorption of fatty acids by enterocytes and microbiota of large intestine].

The decreasing of content of animal, palmitic milk fat (butter) by means of its substitution with vegetable, oleic, palmy oil in food of adults optimal by its quantity is physically chemically and biologically substantiated. In oleic palmy oil higher content of oleic mono unsaturated fatty acid and oleic triglycerides than in creamy fat is established. The biologic availability of palmitic unsaturated palmitic acid in the form of free fatty acid is decreased at its absorption by enterocytes of small intestines is detected. There are no transforms of mono unsaturated acids in palmy oil in contrast with hydrogenated margarines. In palmy, oleic oil there is not enough of short-chained fatty acids (C4-C6) and it has no taste quality and it has low level of unsaturated fatty acids and factually it is lacking of ω-6 polyunsaturated fatty acids. However, it is compensated in case of availability offish and sea products in food. If adults, especially older ones, will refuse to consume creamy fat and decrease intake of products with high content of palmitic unsaturated fatty acid and palmitic triglycerides (beef, sour cream, fatty cheeses) it'll positively impact their health. The refusal from these products is a real step in prevention of metabolic pandemic (atherosclerosis and atheromatosis, metabolic syndrome, resistance to insulin, obesity). There are still large number of people who at optimal amount of food retain in vivo increased amount of exogenous, endogenously synthesized from glucose palmitic unsaturated fatty acid in the form of unesterified fatty acids (syndrome of resistance to insulin) and increased content of palmitic triglycerides.

[The positional isomers of palmitic and oleic triglycerides, lipolysis, conformation of apoB-100, formation of apoE/B-100 ligand and absorption of lipoproteins of very low density by insulin-dependent cells under action of statins.]

The study was carried out to trace back quantitative alterations of positional isomers, oleic and palmitic triglycerides and individual fatty acids in blood serum. After two weeks of taking of Simvastatin (40, 80 mg), analysis of blood serum established decreasing of content of Рhosphatidylcholine and more reliable decreasing of amount of non-etherized alcohol cholesterol. No alterations of content of particular fatty acids were detected. In apoB-100, Рhosphatidylcholine and non-etherized alcohol cholesterol form polar mono-layer; it covers mass of oleic and palmitic triglycerides that was bound by apoB-100 in oleic and palmitic lipoproteins of very low density disturbing bio-availability of substrate (oleic and palmitic triglycerides) for post-heparin lipase. This very pool of non-etherized alcohol cholesterol is inhibited by statins activating lipolysis in oleic and palmitic lipoproteins of very low density. In blood was detected amount of palmitic positional isomersin oleic and palmitic triglycerides (palmitoyl-palmitoyl-palmitate and palmitol-palmitoyl-oleate) and oleic positional isomers (oleyl-oleyl-palmitate and oleyl-oleyl-oleate). The significant difference was marked in content of positional isomers both of oleyl-oleyl-oleate and oleyl-oleyl-palmitate; their content is less in blood of patients of experimental group as compared with healthy people. In case of treatment with Simvastatin (40 mg) the level of palmitic positional isomers and palmitol-palmitoyloleate is decreased. In case of treatment with Simvastatin (80 mg) significant decreasing of positional isomers-palmitolpalmitoyl-oleate and oleyl-oleyl-palmitate as compared with data before treatment. The detection of content of positional isomers triglycerides permits: a) to characterize disorders of metabolism of palmitic fatty acid, oleic fatty acid, oleic and palmitic triglycerides and oleic and palmitic lipoproteins of very low density of the same name; b) to form individual diet therapy; c) to obtain objective information concerning compliance of prescribed recommendations b y patient. The basis of primary prevention of atherosclerosis and atheromatosis is in decreasing up to physiological level in food the content of longchained saturated fatty acids mainly palmitic acid.

Correction of Mitochondrial Enzyme Activities in the Skeletal Muscles of Old Rats in Response to Addition of Olive Oil to the Ration.

Activities of mitochondrial electron transport chain enzymes NADH-CoQ oxidoreductase (complex I), cytochrome C-oxidase (complex IV), and citrate synthase were measured by spectrophotometry in m. quadriceps femoris homogenate from old rats receiving olive oil with the ration. Reduced activities of complexes I and IV in old animals were restored to the level of young animals after 6-week consumption of olive oil. Activity of citrate synthase did not change with age. Positive effect of olive oil on fatty-acid composition of the muscle tissue in old animals was demonstrated. The content of summary monounsaturated fatty acids, reduced with aging, and of summary polyunsaturated ones, increasing with age, were restored in old rats to the levels virtually not differing from the levels of young animals.

[The titration of double bonds in fatty acids of blood plasma in patients in testing of glucose tolerance].

The article deals with per oral glucose tolerance test applied to 20 patients with arterial hypertension. The blood plasma was analyzed to detect content of individual fatty acids, double bounds, glucose, insulin and metabolites of fatty acids. In patients with different resistance to insulin content of non-etherized fatty acids decreased approximatively up to 3 times. Without insulin resistance secretion of insulin in 2 hours after glucose load increased up to 3 times and content of individual fatty acids decreases in greater extent. Under insulin resistance secretion of insulin increases up to 8 times and decreasing of content of fatty acids is less expressed. The decrease in blood plasma of content of oleic and linoleic fatty acids and double bounds reflects effectiveness of effect of insulin--blockade of hydrolysis of triglycerides in subcutaneous adipocytes. The concentration of insulin positively correlates with initial content of palmitic fatty acid in the pool of lipids of blood plasma.

[The content of individual fatty acids and numbers of double bonds, insulin, C-peptide and unesterified fatty acids in blood plasma in testing tolerance to glucose].

The glucose tolerance test demonstrates that content of unesterified fatty acids in blood plasma decreases up to three times and the content of oleic and linoleic acids is more decreased in the pool of fatty acids lipids. Out of resistance to insulin, hormone secretion increases up to three times. The decreasing of level of individual fatty acids occurs in a larger extent. Under resistance to insulin secretion of insulin is increasing up to eight times. The decreasing of level of each fatty acid is less expressed. The effect of insulin reflects decreasing of content of double bonds in blood plasma. The number of double bonds characterizes the degree of unsaturation of fatty acids in lipids of blood plasma. The higher number of double bonds is in the pool of unesterified fatty acids the more active is the effect of insulin. The hyper-secretion of insulin is directly proportional to content of palmitic fatty acid in lipids of blood plasma on fasting. According the phylogenetic theory of general pathology, the effect of insulin on metabolism of glucose is mediated by fatty acids. The insulin is blocking lipolysis in insulin-depended subcutaneous adipocytes and decreases content of unesterified fatty acids in blood plasma. The insulin is depriving all cells of possibility to absorb unesterified fatty acids and "forces" them to absorb glucose increasing hereby number of GLUT4 on cell membrane. The resistance to insulin is manifested in high concentration of unesterfied fatty acids, hyperinsulinemia, hyperalbuminemia and increasing of concentration of C-reactive protein-monomer. The resistance to insulin is groundlessly referred to as a symptom of diabetes mellitus type II. The resistance to insulin is only a functional disorder lasting for years. It can be successfully arrested. The diabetes mellitus is developed against the background of resistance to insulin only after long-term hyper-secretion of insulin and under emaciation and death of ß-cells. The diabetes mellitus type I and not type II is an undesirable outcome of resistance to insulin.

[The fatty acids in blood plasma and erythrocytes in test of glucose tolerance].

The sample of 26 patients with ischemic heart disease and syndrome of insulin resistance was subjected to standard test of glucose tolerance. The content of individual fatty acids was detected using technique of gas chromatography and mass spectrometry. In blood plasma, after 2 hours of post-prandial hyperglycemia, reliably decreased content of C 16:1 of palmitoleic mono fatty acid, C 18:1 oleic mono fatty acid and in a lesser degree C 18:2 linoleic unsaturated fatty acid (p < 0.05). The level C 14:0 of myristic unsaturated fatty acid, C 16:0 of palmitic unsaturated fatty acid and with 18:0 of stearic unsaturated fatty acid, ratio C 16:0/C 16:1 and C 18:0/C 18:1 had no changes: content of both (omega-6 C 20:3 digomo-gamma-linoleic unsaturated fatty acid and essential polyenoic fatty acids remained the same. The significant differences between initial content in blood plasma of palmitic saturated fatty acid and oleic monoenic fatty acid was noted. The alteration in content of fatty acids in membranes of erythrocytes is the most expressed. In erythrocytes reliable (p < or = 0.05) decrease of content of C 16:0 palmitic fatty acid, C 18:0 stearic fatty acid and C 18:1 oleic fatty acid is established. The reliable decrease is noted in content of linoleic unsaturated fatty acid. In erythrocytes, moderate decrease is detected in levels of C 20:4 arachidonic polyenoic fatty acid, C 20:5 eicosapentaenoic polyenoic fatty acid. It is assumed that under post-prandial hyperglycemia insulin regulates metabolism of fatty acids, blocks lipolysis, decreases in cytosol of cells content of oleic and palmitic fatty acids inform of acetyl-KoA and forces mitochondrions intensively oxidate acetyl-KoA formed from pyruvate, from GLU. On surface of membrane, insulin increases number of glucose carriers GLUT4. Hypoglycemic effect of insulin is mediated by regulation first of all of metabolism of fatty acids. Hyperglycemia and insulin are two phylogenetically different humoral regulators. Insulin initiates blockade of lipolysis in adipocytes and positioning on membrane GLUT4. Hyperglycemia passively (activated) increases absorption by cells GLU on gradient of concentration inter-cellular medium--cytosol and synthesis of glycogen.

[The preparation procedure of tests for the gas chromatographic determination of fat acids without preliminary extraction of lipids].

The enhancement of the procedure of quantitative gas chromatographic determination of fit acids in biologic liquids samples is proposed. Instead of the conventional Folch procedure of extraction of lipids with subsequent ablution, concentration and methylation of extracts the direct saponification and methylation of vacuum dried liquid samples (50-200 mkl) can be applied. To compare the effectiveness of the proposed and conventional procedures both of them had been applied to evaluate how converge the results of determination of composition of fat acids in whole blood, blood plasma, packed red blood cells, homogenates of hepatic and muscular tissues. The proposed procedure is applied to determine the characteristics of fat acids composition inpatients with ischemic heart disease.

[The individual fatty acids in blood plasma, erythrocytes and lipoproteins. The comparison of tests results of patients with ischemic heart disease and volunteers].

According to the generally accepted theory, the atherosclerosis is a kind of disorder of metabolism of lipids which chemically are the ethers of fatty lipids with spirits. Hence, the atherosclerosis is fatty acids pathology. In conformity with the biologic classification, among fatty acids it is functionally valid to distinguish saturated fatty acids without double bonds; monoenic fatty acids with one double bond; unsaturated fatty acids with two or three double bonds and polyenic fatty acids with four of six double bonds in chain. The saturated and monenic fatty acids are the substrates for cells to groundwork energy, ATP The unsaturated fatty acids in vivo are needed to form membranes. The polyenic fatty acids are essential since they are precursors of cell synthesis of humoral regulators--eicosanoids (prostanoids and leukotrienes). To clarify the pathogenesis of the "metabolic pandemics" most prevalent in human population, the quantitative determination of individual fatty acids in blood plasma and erythrocytes using gas chromatography technique is needed. It is necessary to evaluate the content of medium chain fatty acids; palmitic and stearic saturated fatty acids; oleic monoenic fatty acid and its transforms--linoleic, linolenic and dihomo-gamma-linolenic unsaturated fatty acids; essential polyenic omega-6 arachidonic, omega-3 eicosapentaenoic and docosahexaenoic fatty acids. The higher is in food the content of palmitic saturated fatty acid, palmitoleic and trans-vaccenic monoenic fatty acids, the more is in patient diet of beef meat and products of fat cow's milk. The higher is ratio of palmitic/oleic fatty acids the lower is the risk of formation of atheromatosis of arteries intima and development of ischemic heart disease and vice versa. The decrease of ratio of omega-3/omega-6 essential polyenic fatty acids is undesirable in prognostic sense. The metabolism of these acids differs and functional activity of omega-3 eicosanoid type 3 is higher In case of deficiency of omega-3 and omega-6 polyenic fatty acids in cells eicosanoids are synthesized from unsaturated dihomogamma-linolenic fatty acid and their influence turns out to be aphysiologic. This condition is a pathogenic foundation of atherosclerosis. There is a diagnostic reason to detect fatty acids in case of diabetes mellitus, obesity, metabolic syndrome and partially arterial hypertension.