[Dynamics of changes of blood inflammatory-oxidative biomarkers in acute coronary syndrome].

Some inflammatory cytokines and parameters of low density lipoproteins (LDL) oxidative modification were studied in blood of 250 acute coronary syndrome (ACS) patients--Siberian inhabitants, men and women with myocardial infarction (MI) or unstable angina on first, tenth and thirtieth days of disease. The inflammatory biomarkers in men and women with MI are: increased concentrations of interleukin (IL)-6, IL-8 and C-reactive protein (CRP), especially on the first day of disease. The most significant inflammatory biomarker of ACS is increased CRP level, especially in women. Oxidative biomarkers in men with ACS are increased basal level of LDL lipid peroxidation (LPO) products and decreased LDL resistance to oxidation. Inflammatory-oxidative biomarkers IL-6, IL-8, CRP and basal level of LDL LPO products are correlated and independently associated with MI.

[Inflammatory-destructive biomarkers of atherosclerotic plaques instability. Study of arterial wall and blood].

Concentrations of tumor necrosis factor, interleukin 1- and its receptor antagonist, IL-6, IL-8, IL-18, IL-2, ligand of CD40 receptor (CD40L), high sensitive C-reactive protein (hsCRP), monocyte chemotactic protein -1, endothelial monocyte activating protein II, adhesive molecules (sICAM-1 and sVCAM-1), matrix metalloproteinase (MMP-3, MMP-7, MMP-9), tissue inhibitor of metalloproteinase (TIMP-1) and endothelin-1 were studied in blood and in coronary artery intima/media of men with coronary atherosclerosis without acute coronary syndrome. Blood levels of hsCRP, IL-8, IL-6 and CD40L were higher, while blood levels of sVCAM and TIMP-1 were lower in men with prevalence of unstable atherosclerotic plaques compared to men with prevalence of stable atherosclerotic plaques in coronary arteries. Blood levels of hsCRP, IL-6 and IL-8 correlated with characteristics of coronary artery atherosclerotic plaques instability. Correlation between hsCRP blood level and hsCRP concentration in coronary artery intima/media material was also revealed.

[Risk of gastric cancer dependent on serological markers of atrophic gastritis: cohort study].

In a prospective study the risk of subsequent gastric cancer (GC) was assessed in persons aged 45-69 over 5 years after the initial testing with a set of serological tests (pepsinogen I, pepsinogen II, gastrin-17, antibodies to Helicobacter pylori). The presence of gastric atrophy markers was a significant predictor of GC in the forthcoming years. Non-invasive techniques may be used in the formation of high-risk groups, followed by GC active surveillance.

[Key laboratory diagnostic biomarkers of coronary atherosclerosis].

Laboratory lipid and lipoprotein biomarkers (total cholesterol - CH, triglycerides - TG, low-density and high-density lipoprotein cholesterol- LDL-CH, HDL-CH, apolipoproteins B and A1 - apoB, apoA1), carbohydrate biomarkers (plasma glucose, basal insulin), high sensitive C-reactive protein (hsCRP) and oxidative biomarkers (basal level of lipid peroxidation [LPO] products in LDL, LDL resistance to oxidation in vitro, oxidative modification of apoLDL and level of LDL lipophilic antioxidants) were studied in 388 men aged 42-70 years: 96 citizens of Western Siberia with angiographically documented coronary atherosclerosis and coronary heart disease (CHD); 292 men of population sample of citizens of Novosibirsk, including 44 men with CHD confirmed by standardized criteria and methods. Significant associations were found of coronary atherosclerosis and CHD with laboratory diagnostic biomarkers like blood levels of HDL-CH, TG, apoB, apoA1, basal insulin, hsCRP and basal level of LPO products in LDL and LDL resistance to oxidation.

[Lipid peroxidation in patients with acute and chronic pancreatitis].

OBJECTIVE: To assess the initial levels of lipid peroxidation (LPO) products in LDL and to study changes in oxidative resistance (OR) of precipitated LDL in acute and chronic pancreatitis patients (OP and CP). METHODS: were examined 39 CP patients and 37 OP patients, and the control group - 7 persons without pancreatic diseases and diabetes mellitus (DM). All CP patients were examined in the acute stage, OP patients - in the first 5 days after admission to hospital. Determination of LPO products in LDL and OR of LDL to Cu2+ - induced oxidation were carried out its own method (Ragino, 1998). RESULTS: In CP patients the initial level of LPO products in LDL was higher (8.9 +/- 1.5 nmol MDA/mg LDL protein) than in those in control group (5.3 +/- 0.8) and in OP patients (5.0 +/- 0.5, p<0.05), and the OR of LDL is significantly lower than those in the control group. In AP and CP patients with permanent pain syndrome in the left hypochondrium initial level of LPO products in LDL is much higher (7.0 +/- 0.6, and 5.0 +/- 0.6, p = 0,016), and OR of LDL after 0, 5, 1 and 2 h incubation with Cu2+ (2 h: 31.7 +/- 1.8 and 23.2 +/- 1.9, p = 0.002) - much lower compared with AP and CP patients without pain. Only CP patients combined with DM type 2 were defined reducing the OR of LDL after 0.5, 1 and 2 h incubation with Cu2+. CONCLUSION: The activity of LPO in the blood and LDL were affected in AP and CP patients.

[Oxidized fibrinogen and its relationship with hemostasis disturbances and endothelial dysfunction during coronary heart disease and myocardial infarction].

The highest oxidative modification of fibrinogen was found in acute myocardial infarction (MI) men and it was 1.26 and 1.56 times higher in comparison with coronary heart disease (CHD) men with anamnesis of MI and with men without CHD, respectively. Increased oxidized fibrinogen level correlated with increased levels of plasma lipid peroxidation products, Willebrand factor, fibrin degradation products, accelerated leukocyte-platelet aggregation and decreased level of plasma NO metabolites. Associations of oxidized fibrinogen with MI and typical parameters of thrombosis and hypercoagulatory hemostasis disturbances and endothelial function were revealed.

[Changes in proinflammatory cytokine and destructive metalloproteinase levels during formation of unstable atherosclerotic plaque].

We studied men with coronary atherosclerosis without acute coronary syndrome and determined typical valuable parameters of inflammatory (tumor necrotic factor, antagonist of receptor to interleukin [IL] 1, IL 6, IL 8, monocytes chemotactic protein 1, endothelial monocytes activating protein II), and destructive (matrix metalloproteinase [MMP] 3, MMP 7, MMP 9, tissue inhibitor of metalloproteinase) processes at consecutive stages of formation of coronary atherosclerotic plaque: "normal intimal tissue --> lipid stain --> early stable plaque --> unstable vulnerable plaque <--> stable plaque with fibrosis", and in 3 types of unstable plaques (lipid type, inflammatory erosive type, necrotic type).

[Cholesterol lowering properties of a complex compound simvastatin with glycyrrhizic acid (simvaglyzin) in experimental models].

A molecular complex of simvastatin (SV) and glycyrrhyzic acid (GA) (at the ratio of 1 : 4), has been synthesized. The complex named "simvaglyzin" (SVG) was stable in aqeous and aqua-alcohol solutions at GA concentrations exceeding 0.2 mM. In vitro SVG acted as uncompetitive inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA reductase (Ki = 94 nM). Appearance of this inhibitory activity is associated with the cytochrome P450-dependent conversion of SVG. The addition of 1 mM methyrapone into incubation medium fully prevented the inhibition of 3-HMG-CoA reductase. SV and SVG (used at 300 nM concentration) inhibited mevalonate synthesis rate by 39.15+/-8.27% and 38.85+/-3.04%, respectively. In vivo SVG showed dose-dependent cholesterol-lowering effect. In rats the cholesterol-lowering effect of SVG used at daily doses corresponding to 66 and 100 mg/kg of SV was equal to the effect of the daily dose 200 mg/kg of SV. The decreases of total cholesterol level in blood serum were 7%, 9% and 8%, respectively. Myotoxicity of those SVG doses estimated by creatine phosphokinase (CPK) activity in blood serum was lower than that of SV. In rats treated with SV the activity of CPK increased by 79% (p<0.01), while in SVG treated rats by 30% and 36% (p<0.05). Any increase of hepatotoxicity markers alanine aminotransferse or aspartate aminotransferase in blood serum was not observed. The data suggest pharmacological synergism attributed to the SV-GA complex formation and elevated safety of the resultant complex compared with the parent compound.

[A procedure for determining the oxidative modification of apolipoproteins in low-density lipoproteins].

The purpose of the present study was to develop a new procedure for estimating the oxidative modification of apolipoproteins in low-density lipoproteins (LDLs). The procedure was developed to use blood serum from 153 males aged 45-65 years, including 69 patients with coronary angiography-verified coronary atherosclerosis and 84 males from a representative sample from Novosibirsk residents of the same age. The new procedure is as follows: a rapid method for isolating serum LDLs, their apolipoprotein (apoLP) protein measurement by the Lowry procedure, their precipitation, a reaction with 2,4-dinitrophenylhydrazine in 2 M HCl solution, by subsequently rinsing in the ethanol:ethyl acetate (1:1) solution, dissolving the precipitate in 8 M urea, and by determining the level of the resultant dinitrophenylhydrazones by spectrophotometry at 363 nm, followed by conversion to LDL concentration of apoLP. The procedure is of informative value for the degree of oxidative LDL modification of apoLP under oxidative stress; it is technically simple, takes little time, and shows a good reproducibility. The values of determined oxidized LDL apoLP by the developed procedure highly positively correlated with the estimates of an oxidized total blood protein fraction and with the values of endothelial dysfunction and did not with the parameters of blood LDL peroxidation. The detected elevated oxidation LDL apoLP in males with coronary atherosclerosis suggests that this index is an additional marker of potentially atherogenic LDL changes.

[Procedure for determination of oxidative modification of plasma fibrinogen].

The purpose of the present study was to develop a new procedure for determining the oxidative modification of plasma fibrinogen. The procedure was developed to use blood plasma from 96 males aged 35-60 years who had cardiovascular diseases: 49 patients with coronary heart disease (CHD), including 16 patients with sub-acute myocardial infarction (MI), and 47 patients with arterial hypertension without CHD. The new procedure is as follows: a rapid fibrin isolating method, a reaction with 2,4-dinitrophenylhydrazine in 2 M HCl solution, by subsequently rinsing in the ethanol : ethyl acetate (1:1) solution, dissolving the precipitate in 8 M urea, and by determining the level of the resultant dinitrophenylhydrazones by spectrophotometry at 363 nm, followed by conversion to the plasma concentration of fibrinogen. The procedure is of informative value for the degree of oxidative fibrinogen modification under oxidative stress; it is technically simple, takes little time, and shows a good reproducibility. The values of determined oxidized plasma fibrinogen by the developed procedure show a high positive correlation with the estimates of oxidized total blood protein fraction and with the values of blood lipid peroxidation. The detected elevated level of oxidized fibrinogen in CHD and MI suggests that this index is a new diagnostic marker of oxidative stress in cardiovascular diseases of atherosclerotic genesis.

[Activity of inflammatory-destructive changes in the process of formation of unstable atherosclerotic plaque].

We studied parameters of inflammatory (tumor necrosis factor a, antagonist of interleukin-1 receptor, interleukin-6 , interleukin-8, C-reactive protein) and destructive (matrix metalloproteinases type 3 and 9, tissue inhibitor of metalloproteinase type 1) processes in dynamics of sequential stages of development of atherosclerotic foci in coronary arteries: unchanged intimal tissue --> lipid stain/streak --> stable young plaque --> unstable vulnerable plaque with inclination to ulceration of rupture --> stable plaque with fibrosis/calcinosis, and in various types of unstable plaques in men with coronary atherosclerosis. Characteristic for unstable plaques parameters of inflammatory activity were elevated levels of interleukins (IL) 6 and 8, C-reactive protein (CRP), of destructive activity -- elevated level of metalloproteinases-9. In inflammatory erosive type of unstable plaques (lowered level of antagonist of interleukin-1 receptor and elevated level of CRP) and in lipid type (elevated levels of IL-6, IL-8 and CRP) inflammatory activity was dominating compared with necrotic type in which dominated destructive activity (elevated level of tumor necrosis factor a and lowered level of tissue inhibitor of metalloproteinase type 1).

[Atherogenic oxidative and structural modification of low density lipoproteins in men with coronary atherosclerosis].

We studied parameters of structural and oxidative modification of low density lipoproteins (LDL) in 80 men aged 35 - 65 years with coronary atherosclerosis verified at coronary angiography in comparison with 80 men of the same age without ischemic heart disease. Biochemical studies included determination of parameters of lipid blood composition by enzymatic methods, subfractional LDL profile by electrophoresis in 2 - 16% gradient polyacrylamide gel, degree of oxidative modification of lipid (lipid peroxidation products) and apoprotein (carbonyl groups) components of LDL by fluorometric and spectrophotometric methods. In men with coronary atherosclerosis we revealed elevated level of small dense LDL and substantially oxidized lipid and apoprotein components of these particles. Independent association of structural and oxidative modification was shown.

[Effect of cytoprotection on the oxidative processes and endothelial function in elderly patients with ischemic heart disease].

The aim of the study was to investigate the effects of cytoprotectional drug Mildronate ("Grindex", Latvia) on some parameters of oxidative processes and endothelial function in elderly patients with coronary heart disease (CHD). One hundred seventeen elderly CHD patients were included into controlled study. The criteria to include the patients into the study were: men and women upwards 60 years old with CHD, with heart failure FC II or III (in accord, NYHA classification), with arterial hypertension (AH), without diabetes mellitus. All patients were randomized into 2 groups: 1) 67 patients 75.4 +/- 0.5 years old were treated with Mildronate 500 mg/day against traditional basal therapy during 12 weeks (the main group) and 2) 50 patients 74.0 +/- 0.6 years old were treated only traditional basal therapy during 12 weeks (the compare group). The blood lipid profile parameters, such as total cholesterol (CH), triglycerides (TG), low density lipoprotein cholesterol (LDL-CH), high density lipoprotein CH (HDL-CH), LDL antioxidant potential (concentration of alpha-tocopherol and retinol in LDL), initial level of lipid peroxidation (LPO) products in LDL, LDL resistance to oxidation in vitro and the blood level of NO metabolites were evaluated before and after 4 and 12 weeks of the study. Effect of Mildronate on the blood lipid profile parameters in elderly CHD patients was not revealed. The initial level of LPO products in LDL was decreased and LDL resistance to oxidation in vitro was increased in main group patients after 12 weeks of study in comparison with the same parameters before the study and with compare group patients (p < 0.05). The blood level of NO metabolites was 1.5-fold higher in main group patients after 12 weeks of study in comparison with the same parameters before the study and with compare group patients (p < 0.05). Thus, in elderly CHD patients the several antiatherogenic effects of Mildronate, such as the decreasing activity of LPO processes in LDL and the increasing synthesis/secretion of blood NO, were revealed.

[Hyperhomocysteinemia in men of Novosibirsk].

Prevalence of hyperhomocysteinemia (HHC) in a sample of male population (n=84, age 50-64 year) of Novosibirsk assessed in the framework of the international project "Determinants of cardio-vascular diseases in the Eastern Europe: multicentral cohort research" during winter-spring period of vitamin deficiency was 50%. In 90.5% of cases HHC was moderate (15-30 micromol/l) and in 9.5% of cases -- medium (30-100 micromol/l). No correlations or independent associations were found between homocysteine blood level and CHD, as well as main risk factors (hyperlipidemia, hypertension, smoking and excessive body weight). No cases of CHD were registered among men with medium HHC. Homocysteinemia correlated positively with age, history of stroke, and negatively -- with alpha-tocopherol concentration in LDL. Men with medium HHC compared with those with normohomocysteinemia had higher systolic and diastolic blood pressure and 29% lower alpha-tocopherol concentration in LDL.

[Oxidized low density lipoproteins and their association with some atherosclerosis risk factors in a population of Novosibirsk men].

Lipid peroxidation products (LPP) and liposoluble antioxidants in low density lipoproteins (LDL), resistance of LDL to oxidation were determined in 305 male inhabitants of Novosibirsk aged 30-70 years. Other methods of examination included questionnaires, anthropometry, registration of blood pressure (BP) and ECG, measurement of total cholesterol (TCH), high density lipoprotein cholesterol (HDLCH), triglycerides (TG). Ten and 90% cut points of percentile distribution of LPP in LDL and parameters of their resistance to oxidation in male population of Novosibirsk were used as regional reference values. Interrelationships were found between LPP in LDL, parameters of LDL resistance to oxidation, levels of antioxidants in LDL, and TCH, TG, HDL CH, smoking, BP and body mass index. Associations between LPP in LDL, LDL resistance to oxidation and TCH, HDLCH, TG, smoking and systolic BP were independent.

[New biochemical methods for evaluation of the oxidative-antioxidative potential of low-density lipoproteins].

Today one of the leading theories in the concept of atherogenesis underlines the key role of oxidized low-density lipoproteins (LDL). This paper describes new original and easy-to-use biochemical methods for evaluating the oxidative-antioxidative potential (LDL resistance to oxidation in vitro and the LDL level of alpha-tocopherol and retinol), as well as the results of their development and testing in 2 groups (62 patients with coronary atherosclerosis and 95 healthy individuals). LDL were isolated from their sera via precipitation with heparin and MnCl2. The precipitated LDL were washed in 0.9% NaCl solution and dissolved in 1 M NaCl solution. For evaluation of the resistance of LDL to oxidation in vitro, the precipitated LDL were incubated at 37 degrees C with 50 microM of copper ions; before and 0.5 h, 1.0 h, and 2.0 h after LDL incubation, TBARS products were determined by the fluorimetric technique after Schun J. et al. (1978). For evaluation of the antioxidative potntial of LDL, the concentrations of alpha-tocopheral and retinol in the precipitated LDL were measured by using the fluorimetric technique described by Taylor S. L. et al. (1976). The new methods were found to be highly reproducible. There was a high positive correlation (r = +0.98 +/- 0.04, p < 0.001) between the oxidative resistance of the LDL isolated from sera by ultracentrifugation and that of the precipitated LDL. There was also a high positive correlation (r = +0.79 +/- 0.09, p < 0.01) between the alpha-tocopherol and retinol levels determined by the fluorimetric technique and the LDL levels measured by the authors' method. The new methods were clinically tested with success. The significantly elevated baseline level of lipid perioxidation products in the precipitated LDL, the low oxidative resistance of the precipitated LDL in vitro, and reduced concentrations of alpha-tocopherol and retinol in the precipitated LDL were recorded in the patients with coronary atherosclerosis as compared to the healthy individuals. These results confirm that LDL oxidative-antioxidative disturbances play an important role in the pathogenesis of coronary atherosclerosis.

[Determination of fractional and subfractional blood lipoprotein compositions with small angle X-ray scattering (a comparison with a biochemical method)]. / Opredelenie fraktsionnogo i subfraktsionnogo sostavov lipoproteinov krovi metodom malouglovogo rentgenovskogo rasseianiia (sravnenie s biokhimicheskim metodom).

Highly accurate and fast method for determination of quantitative and qualitative compositions of serum lipoprotein (LP) fractions and subfractions, using small-angle X-ray scattering on the "Siemens" difractometr, has been developed. The method allows to determine the concentrations of serum cholesterol (CH), triglycerides (TG), high-density lipoprotein (HDL)-CH, low-density lipoprotein (LDL)-CH, phospholipides (FL) and total lipids. It allows to determine the levels of CH, TG, FL in LP subfractions and concentrations of 5 very-low-density lipoprotein, intermedius-density lipoprotein, 3 LDL (large, intermedius and small dense particles) and 5 HDL subfractions. The method is based on the elaborated united model of all LP fractions and subfractions as a result of generalization and analysis of literature data about LP composition and size. New method was proved to be highly reproductive. Strong positive correlation exists between serum levels of CH, TG, HDL-CH determined by enzymatic biochemical and our methods. The method is simple to perform and of high diagnostic value, and can, therefore, be applied to clinical purposes.

[A simple method for estimation of concentration of vitamins E and A in low-density lipoproteins]. / Prostoi sposob otsenki kontsentratsii vitaminov E i A v lipoproteinakh nizkoi plotnosti.

The aim of the study is to work out a new easy method for determination of fat-soluble E and A vitamins concentration in low density lipoproteins (LDL). The method was worked out in the blood serum of 60 healthy men aged 35-55 with monolipidaemia and 55 men aged 45-65 with dislipoproteinemia (DLP). The new method consists of LDL isolation from serum by heparin precipitation method and determination of alpha-tocopherol and retinol concentrations by spectro-fluorimetrical method. The new method is informative for estimation of LDL antioxidant system, easy and fast to perform. The new method features a high reproduction and strong positive correlation between levels of E and A vitamins in the blood serum by spectro-fluorimetrical method and levels of E and A vitamins in LDL by our method. The use of both methods shows reliable reduction in E and A vitamins concentration in the blood serum and in LDL in men with DLP type II compared with E and A vitamins concentration in men with normolipidaemia.

[Blood lipids, oxidative resistance of low density lipoproteins, concentration of fat-soluble antioxidants in older people and longevity in Novosibirsk]. / Lipidy krovi, okislitel'naia rezistentnost' lipoprotinov nizkoi plotnosti, kontsentratsii zhirorastvorimykh antioksidantov u liudei starcheskogo vozrasta i dolgozhitelei g. Novosibirska.

The aim of the study was to evaluate the blood lipid profile, oxidative-antioxidative status of low density lipoproteins (LDL) in old people and long-living people in Novosibirsk. Eighty nine persons from Novosibirsk Veteran House (23 men and 66 women) at the age of 83-105 (average 92.0 +/- 4.0 years old) were included into the study. They were divided into two groups. The group of old people included 12 person at the age of 75-89. The group of long-living people included 77 person at the age of 90-105. The control group consisted of 90 healthy subjects at the age of 36-59. TCH, CH-HDL and TG were measured using biochemical enzyme methods. LDL were isolated from serum by heparin-precipitation method. The initial level of peroxidation lipid (POL) products in LDL and oxidative resistance of precipitated LDL in vitro were evaluated. Concentrations of liposoluble antioxidants (a-tocopherol and retinol) in precipitated LDL were measured by spectro-fluorimetrical methods. The level of CH-HDL was low (< 40 mg/dl) in old and long-living people. In old people the initial level of POL products in LDL was higher in comparison with healthy subjects of control group (p < 0.0001) and long-living people (p < 0.01). In old people and long-living people the oxidative resistance of LDL in vitro was lower (p < 0.0001) in comparison with control group. Moreover, in long-living people the oxidative resistance of LDL in vitro was also lower (p < 0.01) in comparison with old people. The concentration of retinol in LDL was higher only in long-living people compared to the subjects of control group. The concentration of a-tocopherol in LDL was lower in old people and in long-living people (p < 0.05) in comparison with the subjects of control group.

[Galstena efficacy in patients with biliary dyskinesia, chronic cholecystitis and cholelithiasis]. / "Galstena" u patsientov s diskineziei zhelchnovyvodiashchikh putei, khronicheskim kholetsistitom i zhelchno-kamennoi bolezn'iu.

The response of inflammation, bile secretion, lipid metabolism, LDL sensitivity to oxidation was studied in patients with bile duct dyskinesia (BDD), chronic cholecystitis (CC) and cholelithiasis (CL) before and after a course of therapy with galstena. The treatment improved general condition in 90% of the patients, dyspeptic symptoms relieved or stopped. Galstena relieved general or local inflammation, reduced mean levels of total cholesterol and LDL cholesterol, raised HDL cholesterol. In patients with BDD and cholelithiasis galstena did not change normal sensitivity of LDL to oxidation. A galstena course in BDD and CC patients significantly diminished bile lithogenicity and improved gall bladder contractility.