[The contamination under polymerase chain reaction studies: problems and solutions].

The study was carried out to determine risk factors of false positive and false negative results under polymerase chain reaction-analysis of clinical material. The samples with high viral load can be the source of false positive results. The contamination with nucleic acids can occur at any section of polymerase chain reaction analysis. The study data permitted to establish that the most sensitive stage is isolation and purification of nucleic acids especially under manual mode of operation. The detection of positive signal in most samples of one setting indicates total contamination. The cases when only several samples are polluted are special challenge. The presence of sample with high concentration of viral nucleic acid and several samples with low concentration in one setting means necessity of repeated analysis beginning with stage of isolation of nucleic acid. The analysis of curves of accumulation of products of amplification, their forms and positioning on chart is the obligatory stage of polymerase chain reaction study in real time regimen. These actions permit to exclude the readouts of false negative testing results to departments. The study conclusions are equipotent for polymerase chain reaction testing of any nucleic acid targets.

[The clinical biochemistry of hyperlipemia and hyperglycemia. Insulin and metabolism of fatty acids. Hypoglycemic effect of hyperlipemicpharmaceuticals].

The regulation of metabolism of glucose is billions years older than system of insulin and biological function of locomotion (function of motion). Hence hypoglycemic effect of hormone is mediated by alteration of metabolism of fatty acids. The insulin in physiological way deprives mitochondrions a possibility to metabolize ketone bodies, short chain, medium chain and long chain fatty acids and 'forces" them to oxidize glucose which phylogenetically is not an optimal substrate. The relationships between fatty acids and glucose in the Rendle cycle have an effect only on autocrine level (in cell) determining alternation of biological reactions of exotrophia (after food intake) and endotrophia (beyond food intake) in biological function of alimentation (trophology). The most anti-diabetic pharmaceuticals are as insulin hyperlipemic by their mechanism of action. The decrease content of lipid substrates of oxidation in cytosol of cells and mitochondrions "are forced" to oxidize glucose. In these conditions, insulin enhances absorption of glucose by cells through glucose carriers--GLUT4. The derivatives of sulfonil-urea increase secretion of insulin by beta-cells of islets. The biguanidines bond in cytosol covalently and irreversibly ketone bodies taking them away from oxidation in mitochondrions. The fibrates, glitazones, flavonoids and flavones, lipoic tio-fatty acids. The endogenous eicosanoids, derivatives omega-3 and omega-6 of essential polyolefinic fatty acids and conjugated unsaturated fatty acids are the antagonists of receptors of activation of proliferation of peroxisomes. In peroxisomes, they enhance alpha-, beta- and omega-oxidation of all exogenous a physiological fatty acids and excess of palmitic saturated fatty acid forming hypolipidemia in cytozol. The hypolipidemic pharmaceuticals with effect of beta-blocker of oxidation stop absorption of fatty acids by mitochondrions. The omega-3 essential polvolefinic fatty acids, simultaneously with hypolipidemic effect, activate function of GLUT4. In patients of middle age, the diabetes mellitus type II is a symptom of syndrome of atherosclerosis. The reason is that in cells the deficiency of essential polyolefinic fatty acids and is determined by derangement of synthesis of phospholipids and function of GLUT4. It is valid to consider diabetes mellitus primarily as a pathology of metabolism of fatty acids and secondly as a pathology of content of glucose. It is necessary to take into account both under treatment (tactic activities) and strategic program of prevention of diabetes mellitus in population.

[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.

[Low and very low density lipoproteins: pathogenetic and clinical significance].

LDLP and VLDLP have different biological functions: phylogenetically older LDLP transfer FA that serve as substrates for intracellular production of energy and ATP while VLDLP transfer FA--precursors of cell membranes and eicosanoids. The cells absorb LDLP via apoB-100 endocytosis and VLDLP through apoE/B-100 receptors. VLDLP consist of palmitic and oleic VLDLP and LDLP of linoleic and linolenic LDLP. The contribution of LDLP to the development of HLP atherosclerosis and atheromatosis is negligible. LDLP form palmitic and oleic VLDLP with hydrated LDLP density. Blockade of LDLP absorption by apoB endocytosis and deficit of poly-FA constitute the etiological basis of atherosclerosis. Its pathogenetic basis is the excess of palmitic VLDLP with LDPL density in the intercellular space that block absorption of linoleic LDLP with all transferred SC poly-FA. Atheromatosis is clinically and prognostically most significant symptom of atherosclerosis associated with accumulation of ligand-free VLDLP and LDLP in arterial intima of the elastic type as the local pool of interstitial tissue for intravascular pool of intercellular medium. Type 2 diabetes mellitus in aged patients is a symptom of atherosclerosis resulting from SC poly-FA deficit and GLUT4 incompetence. Insulin-dependent cells differ in the degree of insulin resistance. Non-alcoholic fatty liver disease, synthesis of a physiological palmitic TG by hepatocytes and excessive formation of palmitic VLDLP in liver integrate pathogenesis of atherosclerosis and hepatic steatosis. The main pathogenetic factor is the excess of palmitic s-FA and palmitic TG.

[Lipoprotein-associated phospholipase A2 in subjects at low and moderate risk estimated by the SCORE scale].

The aim of the work was to elucidate the relationship between PLA2 content and results of the tests of abnormal lipid transfer in lipoproteins (LP) in subjects at low and moderate risk estimated by the SCORE scale. Another aim was to estimate the diagnostic value of plasma PLA2 content that was determined in 378 subjects (285 women and 93 men) aged 30-64 yr at low and moderate risk (SCORE scale). The patients were divided into groups depending on the age, the number of atheroscleroic plaques (ACP) in carotid arteries (0ACP 1 ACP, more than 1 ACP), enhanced and normal PLA2 levels. PLA2 was measured using PLAC Test Elisa Kits (DiaDexus, USA), with the upper normal limit assumed to be 200 ng/ml. In women, PLA2 levels positively correlated with apoA-1 (main HDLP apoprotein) content (r = 0.51, p < 0.03); in men, PLA2 negatively correlated with TG (r = -0.38, p < 0.01); in subjects with homogeneous ACP PLA2 positively correlated with LP(a) (r = -.38, p < 0.03). Simultaneous rise in PLA2 and LP(a) levels may be a significant risk factor of atherosclerosis and atherothromhosis. Enhanced levels of TG, PLA2, and LP(a) may be the biochemical triad of "soft" plaque formation in the intima.

[The hyperuricosuria in patients with high content of triglycerides: the combination of genetic and environmental factors and tactics of treatment].

The increasing of uric acid level (hyperuricosuria) is regularly detected in blood during the examination of patient with such cardiovascular diseases as arterial hypertension, atherosclerosis, diabetes mellitus, metabolic syndrome and obesity. The hyperiricosuria and hypertriglyceridemia are two independent risk factors, especially for arterial hypertension. The higher level of uric acid combined with hyper-lipoproteinemia (phenotypes) IIa and IIb was noted in 65% of patients. In males, hyperiricosuria was detected more often than in females. In groups with higher content of uric acid, the significant difference between median and quartiles was determined concerning the indicators of height, body mass, triglycerides concentration, beta-lipoprotein fractions content, pre beta-lipoprotein fractions content, apolipoprotein E in blood serum and apolipoprotein B=100 lipoproteins, but not both apolipoprotein C=III and apolipoprotein E in lipoproteins of high density. The increase of concentration of triglycerides and uric acid in blood is the outcome of disorder of metabolism of fat acids and nucleotides under surplus intake of substances with food. The fructose of sweet drinks can be considered as the source of fructose. The fructose is capable to increase the concentration of uric acid The catabolism of nucleotides is under regulatory impact of fructose: dicarboxylic derivatives can provoke increase of uric acid concentration. The treatment of patients with hyper-triglycerideimia, hyperiricosuria and hyperglycemia has to begin from decreasing of triglycerides concentration, dietotherapy and further if it is necessary, to apply the hypolipidemic therapy with fibrates.

[The secretory phospholipase A2 and transport of lipids by lipoproteins in patients with the risk of cardio-vascular pathology (the system "score") of lower and average degree. The diagnostic significance of the test].

The clinical and pathomorphologic data demonstrate that the most frequent cause of cardiac infarction is the formation of "soft" atheromatosis plaques in the intima of arteries. Their rupture results in thrombosis of coronary arteries. The plaques are characterized by higher content of triglycerides. On the basis of the research data, it is possible to validly consider that the detection of secretary phospholipase content A2 conjugated with lipoproteins is the test of systemic inflammatory response. This response is formed under atherosclerosis in vivo as a feedback to the accumulation in the intercellular medium of the endogenic flogogens (initiators of biological reaction of inflammation)--lipoproteins of lower density subclass A. Their utilization in the intima, as a pool of local interstitial tissue, by the resident macrophagocytes transformed from monocytes result in the formation of doth soft and disposed to laceration atheromatosis plaques and the atherothrombosis of coronary arteries and rarer of carotids. Concurrently, the increase of lipoproteins content in blood plasma is supposed to be the test of proliferation of cells in vivo, the smooth muscle cells of medium in particular. The simultaneous detection of content of secretory associated with lipoproteins phospholipase A2 and lipoprotein (a) can be considered as a valid risk factor of atherosclerosis and atherothrombosis--atheromatosis of intima of arteries with the formation of "soft" plaques in the intima, their laceration and thrombosis of coronary arteries and clinical presentation of cardiac infarction. The diagnostic triad of formation of soft plaques in the intima can be composed of the higher level of triglycerides, the content of protein of phospholipase A2 and lipoprotein (a).

[Clinical and laboratory detection of phenotypic characteristics in patients with high hypertriglyceridemia].

The purpose of the study was to define the values of clinical and biochemical (phenotypic) differences in 2 groups of patients: 1) those with moderate (< or =4.5 mmol/l) blood triglyceride (TG) levels and 2) those with high (more than 4.5 mmol/l) blood TG levels and to reveal significant parameters of a diagnostic algorithm for primary and secondary forms of hypertriglyceridemia (HTG). Ninety-six (54%) patients females) with a TG level of more than 2.3 mmol/l were examined. The age was 12 to 71 years (median [quartiles] 50 years [41-61 years]); women accounted for 54%. The patients had the following diseases: coronary heart disease (CHD) (44.89%), myocardial infarction (13.5%), arterial hypertension (87.9%). The diagnosis of HTG included an algorithm for the clinical, biochemical, and clinicogenealogical examination of patients. Biochemical blood analysis involved lipoprotein parameters: cholesterol, triglycerides, low-density lipoprotein cholesterol, lipid electrophoresis, apolipoproteins Al, B-100, E, and C-III. The data were processed using the statistical packages STATISTICA 6.0 and SPSS 17.0. Comparison revealed no age- and gender-related differences in the parameters between Groups 1 and 2 There was a significant correlation between the high levels of TG and the following indicators: total cholesterol, chylomicrones, lipoprotein(a), LP-E , LP B:E, LP C-III4, and LP C-III, smoking (a risk factor) and with the indicators of other metabolic disturbances--total C, chylomicrones, lipoprotein(a), LP-E-total, LP B:E, LP-C3-total, and LP-C3, which determined the impact of nutrition had a hereditary predisposition through the polygenic mechanisms of gene expression under the influence of a number of factors. Pancreatitis was found to be implicated in the development of HTG. Higher TG levels correlated with the parameters, the diagnosis of which reveals additional metabolic disturbances via environmental and polygenic mechanisms

[Diagnostics of moderate and high hypertriglyceridemia in patients in polyclinic practice: primary and secondary lipid metabolic disorders].

AIM: To define clinical and biochemical differences in groups of patients with moderate (< or =4.5 mmol/l and high (more than 4.5 mmol/l) blood triglyceride (TG) levels. To define the markers of biochemical and lipid parameters that could specify an algorithm for the differential diagnosis and treatment of different forms of hypertriglyceridemia. SUBJECTS AND METHODS: Patients (96 (54%) females) aged 12 to 71 years (median 50 years; quartiles, 41-61 years) with a TG level of more than 23 mmol/l and the following diseases: coronary heart disease (CHD) (44.8%), myocardial infarction (13.5%), arterial hypertension (87.9%), xanthomas (36.5%), and a family history of diseases (51%). The diagnosis of hyperlipidemia included a classical algorithm for clinical, biochemical, and clinicogenealogical examinations. Extended biochemical blood analysis, the determination of lipoprotein cholesterol (C), TG, low-density lipoprotein C, lipid electrophoresis, and assay of apolipoproteins A1, B-100, E, and C3 were made. RESULTS: The groups with moderate (< or =4.5 mmol/l and high (more than 4.5 mmol/l) blood triglycerides showed no differences in age and gender, systolic and diastolic blood pressures, the incidence of coronary heart disease, arterial hypertension, peripheral artery atherosclerosis, cardiac arrhythmias, and xanthomas. There was a significant correlation of high TG levels with smoking (a risk factor) and with the indicators of other metabolic disturbances--total C, chylomicrones, lipoprotein(a), LP-E-total, LP B:E, LP-C3-total, and LP-C3, which determined the impact of nutrition (and the development of pancreatitis), but also had a hereditary predisposition through the polygenic mechanisms of gene expression under the influence of a number of factors. CONCLUSION: Higher TG levels correlated with the parameters, the diagnosis of which makes it possible to reveal additional metabolic disturbances via environmental and polygenic mechanisms.

[Treatment of patients with hypertriglyceridemia].

AIM: To assess a hypolipidemic effect and tolerance of phenofibrate (lipantil 200M) in different variants of treatment in patients with combined hyperlipidemia having moderate and high levels of triglycerides (TG). MATERIAL AND METHODS: A total of 26 patients (16 males, 10 females; age 32-67, mean age 55.1 years) received lipantil 200M (phenofibrate) in a dose 200 mg for 12 weeks either as monotherapy or in combination with statins (or fibrate) or plasmapheresis. Clinical, biochemical and blood lipid parameters were studied at baseline and after 12-week treatment. Baseline mean value and standard deviation of a blood cholesterol level was 8.3 (15.4) mmol/l, TG--7.2 (70.5) mmol/l). RESULTS: A mean cholesterol level decreased by 26.1%, TG--by 64.4%. Correlation with initial values was found. Systolic and diastolic blood pressure diminished significantly. No significant changes were found in the levels of AST, ALT, GGT, CPK, alkaline phosphotase, total bilirubin, creatinine. Before the treatment blood glucose was elevated in 14 patients, after 12-week therapy it remained elevated in 10 patients. CONCLUSION: Lipantil 200M (phenofibrate) had a good hypolipidemic effect in patients with combined hyperlipidemia in different baseline levels of cholesterol and TG elevation and in different variants of treatment. Significant side effects were not registered.

[Effect of atorvastatin therapy on the level of secretory phospholipase A2 group IIA and on the modification of low density lipoproteins in patients with ischemic heart disease].

We studied effect of atorvastatin on secretory phospholipase A2 group IIA (sPLA2-IIA) in blood serum of patients with ischemic heart disease (IHD), lipid composition of low density lipoproteins (LDL) and process of modification of LDL induced by sPLA2-IIA in 20 patients taking 20 mg/day of atorvastatin for 3 months. In patients with initially high level of sPLA2-IIA ( > 8 mcg/l) its concentration significantly decreased. Amount of total cholesterol, triglyceride, lecithin, and lysolecithin remained unchanged, however in equimolar relations there occurred decrease of amount of total cholesterol and increase of cholesterol esters. At incubation of LDL, extracted from patient s plasma before initiation of the study, with human sPLA2-IIA from cardiac myxoma, 3.5 nmol of lysolecithin per 1 mg of LDL protein was formed while at incubation of LDL of same patients, extracted after 3 months of atorvastatin administration, amount of lysolecithin was 1.54 nmol/mg LDL protein. Thus atorvastatin therapy causes lowering of sPLA2-IIA in patients with initially high blood level of the enzyme and to a great extent precludes sPLA2-IIA induced LDL modification.

[Double bonds of fatty acids, alcohols glycerol, cholesterol and nonpolar serum lipids. Diagnostic value of hypercholesterolemia]. / Dvoinye sviazi zhirnykh kislot, spirty glitserin, kholesterin i nepoliarnye lipidy syvorotki krovi. Diagnosticheskoe znachenie giperkholesterinemii.

A method for evaluation of double bonds (DB) by ozone titration in a pool of serum lipids during extraction after Folch is developed. A method for measuring the weight of nonpolar lipids in apoB-100-lipoproteins (LP) with a hydrophobic fluorescent probe K-37 was used. Cholesterol and glycerol alcohols were measured in the serum using Cholesterol and Triglycerides kits (Hoffmann la Roche). Twenty-four patients with myocardial infarction, 29 coronary patients, and 37 healthy children were examined. The level of cholesterol alcohol reflects the number of double bonds in fatty acids (FA) and hence, the content of essential polythenic FA in the serum. The higher the level of cholesterol alcohols the greater the pool of essential poly-FA, circulating in the blood as nonpolar cholesterol polyesters within low density lipoproteins (LDL). Presumably, the higher the level of cholesterol LDL, the more pronounced is the deficit of essential poly-FA and the greater is the compensatory activation of the synthesis of endogenous poly-FA. A positive correlation between the content of cholesterol alcohol in the serum and weight of nonpolar lipids in apoB-100-LP was noted.

[Diagnostic significance of albumin transporting characteristics and blood level of troponin T in myocardial infarction]. / Diagnosticheskoe znachenie transportnykh svoistv al'bumina i soderzhaniia v krovi troponina T pri infarkte miokarda.

Serum albumin binding capacity (ABC) was studied using K-35 hydrophobic fluorescent probe in order evaluate the passive transfer and absorption of saturated fatty acids (s-FA) by the cells. ABC decreases on day 3 of myocardial infarction and remains low till discharge and is paralleled by hyperTG. We believe that ABC decrease reflects activation of passive transfer and absorption of s-FA by the cells during simultaneous blocking (presumably by acute phase proteins) of active apoE/B-100 receptor endocytosis of s-FA in the form of TG as VLDL remnants. Decrease of ABC reflects a later than erythrocyte sedimentation rate and acute phase proteins normalization of metabolic consequences of inflammation. ABC helps indirectly evaluate the blocking of active (receptor) absorption of s-FA in the form of TG and activation of passive absorption of s-FA from associations with albumin by the cells.

[Proportion of alcohols of cholesterol, glycerol and double bonds of fatty acids in blood serum and lipoproteins. Diagnostic significance of hypercholesterolemia]. / Vzaimootnosheniia spirtov kholesterina, glitserina i dvoinykh sviazei zhirnykh kislot v syvorotke krovi i lipoproteinakh. Diagnosticheskoe znachenie giperkholesterinemii.

A new method in quantitative determination of double bonds (DB) in the pool of fatty acids (FA) has been worked out by ozone titration after extraction of lipids according to Folch. A correlation was observed among DB content, the level of cholesterol alcohol (CS, r = +0.612; p < 0.001) and the level of glycerol (GL) alcohol (r = +0.392; p < 0.01). The grown-ups with ischemic heart disease and children have a constant ratio (5.05 +/- 0.18) DB:CS ratio.