Stable Metabolites Of Nitric Oxide Research Articles

PROGNOSTIC SIGNIFICANCE OF ENDOTHELIAL DYSFUNCTION MARKERS IN ARTERIAL HYPERTENSION

Aim. Assessment of prognostic significance of endothelial dysfunction markers: stable metabolites of nitric oxide (NOx), von Willebrand factor (vWF), endothelin-1 (E1), homocysteine and tissue plasminogen activator (tPA) in essential hypertension (EAH) patients not taking antihypertension therapy systematically.Material and methods. Totally, 12 EAH patients investigated (45 males, 79 females) (mean age 51,4±6,5 y. o., mean duration of AH 7,9±7,3 y.). Concentration of NOx in plasma was measured by spectrophotometry, and of vWF, homocysteine, E1 and tPA — by immune enzyme assay.Results. By the increase of SCORE risk level, there was significant increase of concentrations of NOx, E1, homocysteine and vWF in EAH patients (p<0,05), there were no changes in tPA levels (p>0,05). In 8 (8±1,1) years after baseline assessment, 115 patients were assessed second time. Of those 13 (11,3%) had cardiovascular events (CVE) and 5 (4,3%) died. By single factorial regression, the rate of CVE in EAH patients relate to homocystein level (р=0,01), NOx (р=0,001) and vWF (р=0,001). By multifactorial analysis, prognostic statistical significance is found for NOx (relative risk (RR) =3,8, р=0,006) and vWF (RR =3,5, р=0,005). In ROC-analysis there were found threshold levels of NOx (>46,6 mcM/L, AUC =0,863) and vWF (>1,68 mg/dL, AUC =0,738), the increase of which is followed by CVE development risk for the levels of NOx >46,6 mcM/L 3,8 times (sensitivity 81,9% and specificity 65,8%), vWF >1,68 mg/dL — 3,5 times (sensitivity 74,3% and specificity 62,7%). Combination of the parameters point on the risk increase up to 6,5 times (р=0,00007).Conclusion. NOx with the threshold of >46,6 mcM/L (RR =3,8) and vWF >1,68 mg/dL (RR =3,5) do show independent prognostic value for 5-year CVE risk assessment in EAH patients that can be applied as an additional method for risk stratification to estimate a group for more aggressive therapy and CVE prevention.

Improvement of fetus growth restriction diagnostics in pregnant women by means of biochemical markers that characterize the disorder of stress-adaptation

Object of the work. Regulatory and adaptive processes of the system “mother –placenta – fetus” state determination and detection of pregnant women adaptive capabilities specific features in case of fetus growth restriction based on a number of the blood plasma biochemical indicators.Materials and methods. Markers of oxidative stress were detected spectrophotometrically in the blood plasma according to generally accepted methods: oxidative modification of proteins, stable metabolites of nitrogen oxide, L-arginine, malondialdehyde, thiol compounds and reduced glutathione. Statistical processing of data was made by methods of variational statistics with Microsoft Office Excel 2010, Statistica 6.0.Results. The use applicability of the number of biochemical markers which are predictors of perinatal complications has been scientifically substantiated. Using biochemical diagnostic technique it has been shown that fetus growth restriction syndrome is accompanied by disorders of regulatory and adaptive processes of the system “mother –placenta – fetus”. In particular, the level of proteins and lipids oxidative modification increases, nitric oxide synthase activity decreases and reserves of L-arginine and thiol compounds decrease in the blood plasma.Conclusions. Comparative analysis of a number of biochemical markers determination results of stress-realizing system activity in pregnant women with fetus growth restriction and in women with a physiological pregnancy has revealed a significant (P ≤ 0.05) increase in markers of protein oxidative modification blood levels (the level of dinitrophenylhydrazones of aliphatic aldehyd of basic amino acid residues and dinitrophenylhydrazones of carbonyl compounds of basic amino acid residues in the spontaneous sample increased by 14.2 % and 16.3 %, respectively, and in the stimulated sample by 46.6 % and 43.0 %), the level of malondialdehyde increased by 42.9 %. Reduction of the stress-limiting system activity in pregnant women with fetus growth restriction is indicated by a decrease in the content of stable metabolites of nitric oxide in blood plasma by 51.2 %, reduction of L-arginine content by 55.5 %, glutathione by 45.9 % and free thiols by 52.4 %. The aforementioned changes aggravate fetal trophism and can effect negatively on the process of cervix preparing for delivery.

The use of immunomodulators, antioxidants and hepatoprotectors for the correction of the liver, erythrocites and the immune system disorders in chronic ethanol intoxication

The effectiveness of three various combinations of an immunomodulator with an antioxidant and a membrane protector in the correction of metabolic and immune disorders has been studied in the experiment under 60-days ethanol intoxication. The development of such biochemical syndromes of the liver damage as cytolysis, intrahepatic, intracellular cholestasis, toxic liver damage by necrotic type, insufficiency of synthetic processes and inflammatory has been revealed. Oxidative stress development and the activation of lipid peroxidation on the systemic (blood plasma) and local level (erythrocytes) have been established. Suppression of adaptive immunity formation and phagocytic capabilities of neutrophils under the increase in their oxygen-dependent activity has been determined, which indicates the presence and possible progression of the inflammatory process at the systemic level. A disorder of erythrocytes metabolic activity, a decrease in stable metabolites of nitric oxide detected in blood plasma been revealed, indicating its uncompensated consumption, causing vasoconstriction and thrombosis, which can additionally arise due to the established increase in the prothrombin index. Combined use of "Longidasa", "Mexicor", "Essentiale forte N" or "Glutoxim", "Mexidol", "Heptral" was more effective in the correction of immune-metabolic disorders in chronic alcohol intoxication than "Hepon", "Hypoxen" and "Phosphogliv".

THE STATE OF ENDOTHELIUM-DEPENDENT VASOREGULATION IN THE DYNAMICS OF THE EFFECT OF A NON-DRUG THERAPY COMPLEX IN PATIENTS WITH PULMONARY ARTERIAL HYPERTENSION

After complex treatment with the use of ozonotherapy and bischofite electrophoresis on the background of standard therapy, the parameters of diastolic function of the right ventricle, mean pulmonary arterial pressure, the level of stable metabolites of nitric oxide, endothelium-dependent vasoregulation and psychoemotional status in patients with chronic obstructive pulmonary disease complicated by the development of chronic pulmonary heart were improved against standard therapy.

MORPHOLOGICAL AND FUNCTIONAL STATE OF ENDOTHELIUM AT PATIENTS WITH CORONARY ARTERY DISEASE AND CHRONIC GLOMERULONEFRITIS

All over the world the increase in quantity of patients with vascular complications of chronic inflammatory diseases of kidneys is marked. The interrelation of morphological and biochemical markers of atherosclerotic lesions were studied in 41 patients with coronary artery disease (CAD) (group 1) and 62 - with CAD and chronic glomerulonephritis (CGN) (group 2). An extracranial carotid artery (CA) status was investigated by duplex ultrasonography. VE-cadherin content determined by ELISA. States of the antioxidant system was evaluated by the concentration of malondialdehyde (MDA) and sulfhydryl groups (SH-groups) in the blood plasma by the photometric method. The levels of nitric oxide stable metabolites – NO2 and total (NO2+NO3) in plasma was determined by the photometric method. There was found that the frequency of CA plaques detection in groups 1 and 2 did not differ (27,6% and 31,0%, respectively), but patients of group 2 were reliably younger. Pronounced VE-cadherin increase (3,6-fold, p <0,01) at the background of the deterioration of renal function (2,24 -fold GFR decrease, and urinary protein excretion reaches (0,200,98) g / day) was observed in group 2. The content of NO2 in group 2 was significantly lower than in controls, and total (NO2+NO3) was increased as compared with control (2,40-fold, P <0,01), and with patients of group 1 (in 1,86-fold, p <0.05). Increase of MDA level in patients of group 2 (2,61-fold, P<0,01) accompanied by reduction of SH groups content – by 25,2% (P <0.05) compared with control. Thus, in patients with combined pathology revealed the most pronounced disorders of endothelial integrity and function, which was worsening by imbalance in the pro-antioxidant system. Accelerated atherosclerotic vascular changes in these patients are probably related to the chronic inflammatory process.

PATHOGENETIC SUBSTANTIATION OF L-ARGININE APPLICATION AT COMBINED CLINICAL COURSE OF NON-ALCOHOLIC STEATOHEPATITIS AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE TAKING INTO ACCOUNT POLYMORPHOUS VARIANT G894T OF THE GENE OF ENDOTHELIAL NO-SYNTHASE

Objective. The aim of the study was to investigate the functional state of the endothelium and liver, carbohydrate metabolism and lipid spectrum of blood, depending on the polymorphism of the eNOS gene (G894T / G1917T) in patients with non-alcoholic steatohepatitis combined with chronic obstructive pulmonary disease, in the dynamics of complex treatment with baseline L -arginine.Material and methods. The efficacy of administering L-arginine to 60 patients with non-alcoholic steatohepatitis in combination with chronic obstructive pulmonary disease, taking into account the polymorphism of the eNOS gene (G894T/G1917T), was studied by examining the content of endothelin-1, the number of circulating desquamated endotheliocytes, sVCAM-1, stable metabolites of nitric oxide, total cholesterol, triglycerols, low density lipoprotein cholesterol, very low density lipoprotein cholesterol and high-density lipoprotein cholesterol, the presence of glucose and glycosylated hemoglobin in the blood on an empty stomach.Results. The use of L-arginine hydrochloride (intravenously) followed by administration of L-arginine aspartate (per os) against a background of baseline therapy of non-alcoholic steatohepatitis in patients suffering from chronic obstructive pulmonary disease with GG genotype and with GT + TT genotype (with dose adjustment and treatment duration) contributes to a significant decrease of endothelin-1 the level, sVCAM and the number of circulating desquamated endotheliocytes, while the level of nitrates / nitrites in the blood serum increases. At the same time there is a positive dynamics of glucose, glycosylated hemoglobin, total bilirubin, aminotransferase activity, γ-glutamyltranspeptidase, triglycerol, low and high density lipoprotein cholesterol in blood serum.Conclusions. The use of L-arginine in the complex treatment of patients with non-alcoholic steatohepatitis combined with chronic obstructive pulmonary disease leads to a more pronounced improvement in the functional state of the liver and endothelium, carbohydrate metabolism and lipid profile in the group of patients with GG genotype and in the group of patients with GT + TT genotype, which was used to adjust the dose and duration of treatment compared to the group of patients with GT + TT genotype, who did not undergo treatment correction and to whon L-arginine was not prescribed.

Endothelial immune activation and functional state in patients with hypertensive disease

To investigate the nitric oxide (NO) system, the activity of endothelial NO synthase (e-NOS) and inducible NO synthase (i-NOS) in relation to the levels of tumor necrosis factor-α (TNF- α) and its soluble receptor type I (sTNFRI) depending on the grade of hypertensive disease (HD). A total of 317 patients, including 284 patients aged 30 to 65 years (mean age 54.7±0.58 years) with Grades 1-3 HD who had not previously received regular antihypertensive therapy and 33 apparently healthy individuals, were examined at Kharkov City Clinical Hospital Eleven. Immune activation was judged by the serum levels of the proinflammatory cytokine TNF-α in all the examinees and y those of sTNFRI, which were measured by enzyme immunoassay. To study the endothelial functional state, the level of stable end metabolites of nitric oxide, that of S-nitrosothiol and the activity of NO synthases were biochemically determined in 100 patients from this group. A control group consisted of 16 apparently healthy individuals. There were increases in the circulating levels of proinflammatory cytokines (TNF-α, sTNFRI), the content of S-nitrosothiol and the activity of i-NOS. At the same time, there were decreases in the activity of e-NOS and the level of end nitric oxide metabolites, such as nitrites and nitrates. In patients with HD, the end metabolites of nitric oxide decrease, which indirectly shows a reduction in its vasoactive part, and the stable NO metabolite S-nitrosothiol increases. This is associated with enhanced NO oxidation under conditions of oxidative stress and endothelial dysfunction. The higher amount of S-nitrosothiol in the examinees may be associated with increased i-NOS. The immunoinflammatory activation mediated by the proinflammatory cytokines, particularly by the enhanced activity of TNF-α and sTNFRI, has been proven to play a role.

Abstract P185: Prognostic Value of Endothelial Markers in Hypertensive Nondiabetic Patients

Aim: to study the prognostic value of the serum levels of stable nitric oxide metabolites (NOx), endothelin-1 (E1), homocystein (HC), von Willebrand factor (Wf), tissue plasminogen activator (tPA), in hypertensive nondiabetic patients without cardiovascular disease. Methods: 124 patients with hypertension (HTN), without diabetes were enrolled, the mean age was 51.4±6.5y, the mean duration of HTN was 8.5±7.6 y, 36 % were males, 35% were smokers, 54% were obese. The cardiovascular risk was estimated according to the SCORE (Systematic Coronary Risk Evaluation) system. Plasma NOx levels were determined by spectrophotometry, E1, HC, Wf, tPA levels by enzyme immunoassay. The follow-up period was 8±1.1y. Statistical analysis was done using the SPSS 11.1 software. Results: On initial investigation all patients were stratified according to the SCORE risk levels. The increase of SCORE risk levels were associated with the elevation of serum NOx, E1, HC, and Wf concentrations. The mean plasma NOx concentration increased along with the SCORE risk level: it was 36.4±12.7 in low risk, 41.1±15.7 in moderate risk, 44.9±19.8 in high risk, and 50.4±19.1 umol/L in very high risk categories (<0.05 between the low and high risk groups). The tPA levels did not differed in relation to the SCORE risk estimates. The SCORE risk level correlated with serum concentrations of NOx (r=0.74, p=0.0001), HC (r=0.54, p=0.0001), and Wf (r=0.43, p=0.001). Conclusions: The stable nitric oxide metabolites, homocystein, and von Willebrand factor are the markers of oxidative/nitrosative stress severity. Our results are indicative that this substances may be of prognostic value in hypertensive nondiabetic patients.

Experimental studies on possible regulatory role of nitric oxide on the differential effects of chronic predictable and unpredictable stress on adaptive immune responses

The present study was designed to investigate the effects of chronic predictable stress (CPS) and chronic unpredictable stress (CUS) on immunological responses in KLH-sensitized rats and involvement of NOergic signaling pathways mediating such responses. Male Wistar rats (200–250g) were exposed to either CPS or CUS for 14days and IgG antibody levels and delayed type hypersensitivity (DTH) response was determined to assess changes in adaptive immunity. To evaluate the role of nitric oxide during such immunomodulation, biochemical estimation of stable metabolite of nitric oxide (NOx) and 3-nitrotyrosine (3-NT, a marker of peroxynitrite formation) were done in both blood and brain. Chronic stress exposure resulted in suppression of IgG and DTH response and elevated NOx and 3-NT levels, with a difference in magnitude of response in CPS vs CUS. Pretreatment with aminoguanidine (iNOS inhibitor) caused further reduction of adaptive immune responses and attenuated the increased NOx and 3-NT levels in CPS or CUS exposed rats. On the other hand 7-NI (nNOS inhibitor) did not significantly affect these estimated parameters. The results suggest involvement of iNOS and lesser/no role of nNOS during modulation of adaptive immunity to stress. Thus, the result showed that predictability of stressors results in differential degree of modulation of immune responses and complex NO-mediated signaling mechanisms may be involved during responses.

Success in the acquisition of Bombyx mori sperm motility is influenced by the extracellular production of nitric oxide (NO) in the presence of seminal fluid nitric oxide synthase (NOS)

A trypsin-like protease called initiatorin is known to initiate sperm motility in the silkworm, Bombyx mori, but little is known about the signaling events leading to sperm flagellar beating. The aim of this study was to investigate whether this mechanism of sperm motility activation involves the signaling transmitter nitric oxide (NO). NO is produced from the amino acid L-arginine by the enzyme action of nitric oxide synthase (NOS; EC 1.14.13.39). Simple treatment of quiescent sperm with an NO donor (SNAP or NOC7) in vitro did not lead to activation of motility. Nevertheless, initiatorin- or trypsin-induced motility was blocked by pretreatment of sperm with either the NOS inhibitor L-NAME or NO scavenger carboxy-PTIO. These observations suggested that NO may play important physiological roles in the acquisition of sperm motility under the in vitro condition used here. Then, we investigated whether NO synthesis would occur in the spermatophore, a capsule containing spermatozoa that is created by the contents of various male reproductive glands and is the site of sperm maturation. The amounts of NO2− and NO3−, stable metabolites of NO, reached maximum values after enclosure in the spermatophore, a time when apyrene spermatozoa acquire vigorous motility. Moreover, RT-PCR and Western blotting analyses of NOS indicated that it is abundantly expressed in glandula (g.) lacteola of the virgin male ejaculatory duct, from which it is secreted to the seminal fluid and transferred to the female during mating. Previous studies demonstrated that free L-arginine is supplied de novo by a specific proteolytic reaction in which initiatorin participates during spermatophore formation (Osanai et al., 1987c). Based on these results, it can be presumed that the mixing of seminal fluid contents from each male reproductive organ during ejaculation induced NO production outside of the spermatid, and exogenous NO stimulated a signaling pathway involved in the activation of silkworm apyrene sperm.

OXIDATIVE STRESS IN ACUTE SEROUS AND PURULENT PYELONEPHRITIS

THE AIM: To study the state of the proand antioxidant system at the systemic and local level in acute serous and purulent pyelonephritis (ASP, APP). PATIENTS AND METHODS. Oxidative status was studied in 62 patients with ASP and APP, randomized by age, sex, the minimum number of co-morbidities in remission. RESULTS. It was found that the ASP, to a greater extent with the APP on the system (blood plasma, red blood cells) and local level (urine) increases the concentration of stable metabolites of nitric oxide, increases the content of lipid peroxidation products (alkyl hydroperoxides, malonic dialdehyde) and decreases the activity of the antioxidant system (superoxide dismutase levels, catalase, in plasma, urine and erythrocytes). After the comprehensive treatment of the studied indices at ASP are corrected or normalized, and at APP are subject to a minimum correction. CONCLUSION. The results reveal changes of oxidative status occurring in acute serous and purulent pyelonephritis, and also confirm the need for the comprehensive treatment of this disease using drugs with antioxidant and cytoprotective effects.

Level of Nitrogen Oxide and Indicators of Oxidative-Antioxidative Status in the Gingival Tissues, Blood Plasma in Experimental Chronic Gastritis and Duodenitis and Drug Correction

model of chronic gastritis and duodenitis was conducted in 48 immature Wistar rats both gender by intragastric injection of medical bile. Control group consisted of 10 animals. After approximation model of the gastritis and duodenitis rats were divided into five subgroups: the 1st subgroup (before treatment, 9 rats), subgroup 2 (10 animals) received the antioxidant, subgroup 3 (11 animals) – received NO donator, subgroup 4 (10 animals) - received an antioxidant and NO donator, subgroup 5 (8 animals) – received the antioxidant, NO donator and calcium carbonate. In rats with gastritis and duodenitis was observed redistribution of stable metabolites of nitric oxide: decreasing level in the gingival homogenates and increasing in the blood plasma. Level of malondialdehyde and aldehyddehydrogenase in the gingival tissue increased, and in the blood plasma decreased, on a background of catalase activation; content of ketone phenilhydrazone remained in the level of control group. Decreasing markers of oxidation lipids and proteins in the blood plasma, on a background of increasing levels of catalase and nitric oxide metabolites (in 14 times), indicates about a key role of NO in the antioxidant protection of organism in a case of disease.Application of antioxidant could not impact on the indicators of oxidation lipids and proteins. It had been shown decreasing content of the nitric oxide metabolites in the gingival tissues and blood plasma. Drug correction with donator of nitric oxide have to increase markers of oxidation lipids and proteins in the gingival tissues, on a background of high catalase activity and low levels of nitric oxide metabolites. In the blood plasma content of nitric oxide metabolites was higher, which indicated about an active inflammatory process in the stomach and duodenum mucosa. Indicators of the protein molecules fragmentation and malondialdehyde were not differ from those before treatment. Level of catalase was in two times higher, than in the control group, but level of superoxide dismutase was decreased. Complex of antioxidant and donator of nitric oxide helped to stabilise the indicators of oxidation lipids and proteins, although level of nitric oxide metabolites in the gingiva was significantly decreased. Application complex from the antioxidant, donator of nitric oxide and calcium carbonate at the treatment of chronic gastritis and duodenitis restored an oxidative-antioxidative status in the blood plasma and gingival tissues, increasing the production of nitric oxide to a level in the control group.

ENDOTHELIAL DYSFUNCTION IN ESSENTIAL HYPERTENSION IN ADOLESCENTS (CLINICAL AND EXPERIMENTAL STUDY)

Objective. The aim of the study was to evaluate endothelial dysfunction based on the blood concentration of endothelin 1 (ET‑1), nitric oxide (NO), serotonin (5‑HT) in essential hypertension (EHTN) in adolescents and in experimental animal models. Design and methods. The work was carried out in two stages. The clinical part was preceded by the investigation of experimental models of arterial hypertension (HTN). Twenty five immature male SHR rats aged 6–7 weeks and 10 immature male Wistar–Kyoto rats aged 6–7 weeks (control group) were examined. The clinical part of the study included 70 untreated male adolescents (mean age 15,63 ± 0,26 years), among them 24 people had a labile form of EHTN (LHTN), 30 had stable form of EHTN (SHTN), the control group included 16 subjects. All participants underwent 24-hour blood pressure monitoring. The plasma concentration of EN-1 (ET‑1) was determined by enzyme immunoassay, total concentration of stable metabolites of NO, 5‑HT in serum and platelets was measured. Results. The serum levels of ET-1 and 5-HT were higher in HTN groups than in the control group, and in adolescents with SHTN levels of both indices were higher compared to those with LHTN (p < 0,0001). Serotonin level in platelets in HTN groups was lower than in the control group (p < 0,0001). Compared to the control group, NO levels were higher in adolescents with LHTN, but lower in patients with SHTN (p < 0,0001). The changes in the level of ET‑1, 5‑HT and NO in plasma, serum and blood platelets in the experimental animal model were similar to those found in patients with labile and stable HTN. Conclusions. NO, ET‑1 and 5‑HT are early sensitive markers of endothelial dysfunction in EHTN in adolescent males, as well as in the experimental SHR rat model.

ENDOTHELIAL DYSFUNCTIONS AND THEIR CORRECTION IN WOMEN WITH EARLY POSTMENOPAUSE

Aim. To evaluate the nature of the dysfunction of the endothelium in postmenopausal women with climacteric syndrome, peculiarities of its changes during long-term use of a fixed low-dose combination of 17β-estradiol 1 mg/drosperidone 2 mg daily (Е2 1 mg/DRSP 2 mg). Material and methods. A non-randomized, uncontrolled study included 162 women in early postmenopausal period with climacteric syndrome. Patients were divided into 2 groups. Patients of the main group (n=84) received menopausal hormone therapy (MHT) with the fixed combination of Е2 1 mg/DRSP 2 mg. Patients of the control group (n=78) did not receive MHT. The duration of the follow-up was 5.2 years. The endothelium dependent vasodilation (EDVD) of brachial artery was assessed by reactive hyperemia. Endothelial dysfunction (ED) markers - plasma levels of endothelin-1 (ET-1), NO metabolites, asymmetric dimethylarginine (ADMA) and von Willebrand factor antigen (vWF:Ag) - were studied by a highly sensitive quantitative enzyme-linked immunosorbent assay. Results. The examined postmenopausal women with climacteric syndrome had significant increase in the concentration of ET-1 up to 1.0 (0.7-1.3) fmol/ml, increased levels of vWF:Ag up to 0.835 (0.760-0.990) U/ml and reduction in the level of the total content of stable metabolites of nitric oxide (NOx) to 39.2 (35.4-43.7) μmol/l. Disorders of EDVD of the brachial artery were observed in 50 (59.5%) women of the main group and in 45 (57.6%) women of the control group (p=0.7). The relationships between EDVD parameters and levels of ADMA (r=-0.31, p=0.029) and NOx (r=0.31, p=0.002) were revealed. In patients with impaired EDVD, a significant decrease in the level of stable NO metabolites and an increase in ADMA and ET-1 levels were detected. A significant increase in the EDVD of the brachial artery was observed throughout the follow-up period. Besides an increase in NO metabolites level, and decrease in the ET-1 and ADMA levels were found by the 12th month of MHT. Conclusions. Disorders of endothelial function were found in the majority of women in early postmenopausal period, the most marked changes were in patients with impaired vasoreactivity of brachial artery. Favorable changes in vascular endothelium function were observed during long-term MHT with the fixed combination of Е2 1 mg/DRSP 2 mg.

THE STATE OF NITROXIDE PRODUCTION FUNCTION OF ENDOTHELIUM AND LEFT VENTRICLE HYPERTROPHY IN ARTERIAL HYPERTENSION

Aim . To assess levels of stable nitric oxide metabolites (NO) in plasma of arterial hypertension (AH) patients not taking systematic antihypertension therapy, in accordance with the presence and absence of the left ventricle hypertrophy (LVH). Material and methods . Totally, 124 AH patients included: 45 males, 79 females, mean age — 51,4±6,5 y.o., mean duration of AH — 7,9±7,3 y. Controls included 25 healthy persons: 10 males, 15 females; mean age — 48,2±7,8 y.o. Transthoracal echocardiography was done by standard method. Myocardium remodelling types were evaluated by the Ganau A (1992) criteria. Concentration of NO in plasma was measured by spectrophotometry. Statistics was done with Statistica 8.0 software. Results . In AH patients the level of NO was significantly higher than in controls — 43,2±21,7 mcM/L vs 28,3±9,6 mcM/L (p<0,05). There was positive correlation of NO with the fact of AH (r=0,31, p<0,05). In AH patients LVH was diagnosed in 66,7%. In AH with LVH patients concentration of NO was significantly higher than in AH non-LVH — 49,9±19,3 vs 39,8±18,3 mcM/L, respectively, (р<0,05) and in controls. There were direct correlations in AH patients, with NO and myocardial mass index of the left ventricle (LV) (r=0,44, р<0,05), with interventricular septum thickness (r=0,36, р<0,05), posterior LV wall thickness (r=0,44, р<0,05). In the subgroup of AH patients with normal LV geometry there were the lowest NO levels — 35,1±17,3 mcM/L. In subgroups of AH patients with concentric remodeling, eccentric remodeling and concentric LVH, levels of NO were significantly higher than in controls (p<0,05). Higher NO levels were found in AH patients with concentric LVH (53,9±19,1 mcM/L). Conclusion . In AH patients of I-II stages there were raised NO levels. In AH patients with LVH concentration of NO is significantly higher than in non-LVH. There is correlation with myocardium mass index of LV and NO (r=0,44, p<0,05). The highest levels of NO were found in AH patients with concentric LVH.

IMPACT OF CHRONIC ARTERIAL HYPERTENSION ON CLINICAL AND METABOLIC NEONATAL ADAPTATION

Aim. To assess the impact of chronic arterial hyper-tension on clinical and metabolic adaptation of fullterm newborns. Methods. 65 full-term neonates born to women with chronic arterial hypertension were included in the study. 42 neonates born to women with arterial hypertension stage I were enrolled into Group 1, and 23 neonates born to mothers with arterial hypertension stage II were included in Group 2. Levels of renin, angiotensin II, aldosterone, natriuretic peptides, endothelin-1 in umbilical cord blood and peripheral blood were measured with the ELISA. Stable nitric oxide metabolites levels were measured using the Griess assay. Results. Chronic high blood pressure during pregnancy has been found to affect fetal central nervous system and cardiovascular system. Early neonatal period in infants of women with chronic arterial hypertension is associated with a high incidence of perinatal CNS injury, continuing with little change during the first three months of life. The features of the cardiovascular system in neonates of women with arterial hypertension stage II include hypertension and tachycardia at birth, violation of atrioventricular conduction on the first three days of life, open patent foramen ovale up to the first three months of life, whereas neonates born to women with arterial hyper-tension stage I have hypotension on the first day of life. Elevated levels of angiotensin II in neonates born to women with chronic arterial hypertension at birth and reduced levels of endogenous nitrite in the first three months of life suggest the activation of the renin-angiotensin-aldosterone system in the antenatal period, which may cause endothelial dysfunction in early infancy. Conclusions. 1. Chronic high blood pressure in pregnancy is as-sociated with perinatal CNS damage in the newborn, continued during the first three months of life. 2. Stage 1 of hypertension in pregnancy associated with hypotension in newborns on the first day of life; and stage 2 of hypertension in pregnancy associated with hypertension and tachycardia newborn at birth, a violation of his atrioventricular conduction on the 3rd day of life and preservation until the age of three open oval window. 3. Chronic high blood pressure in pregnancy is as-sociated with an increase in cord blood levels of angiotensin II decrease in peripheral blood concentration of endogenous nitrite in the age of three months.

ROLE OF NITRIC OXIDE METABOLITES WITHIN THE IMPACT OF THE SUB-TOXIC SUCCINAMIDES DOSES ON STATE OF HEMOSTASIS.

We investigated the role of changes in the endogenous nitric oxide (NO) metabolism during the influence of succinic acid amides as biotransformation products of an anti-diabetic drug on the state of hemostasis. In experiment with rats, synthetic succinamides were applied in quanti- ties equimolar to the sub-toxic dose of the pharmaceutical substance. We investigated the indicators characterizing the state of platelet and coagulation hemostasis in the blood plasma, the content of the stable NO metabolites and the activity of nitrogen oxide synthase (NOS) in the liver homogenate, blood plasma and urine of rats. We found that sub-chronic succinamides introduction reduced the nitrite and nitrate anions concentration in the blood plasma (by 30-50 and 20-35% resp.), liver (by 16-19 and 14-18%) and urine (by 50-70 and 38-55%). These changes were essentially dependent on the reduction in the NOS activity (by 33%). The studied compounds showed a 1.5 fold increase in the coagulation potential of the blood plasma and cause a 20% boost in the aggregation of thrombocytes. Analysis of the pair correlation coefficients showed positive association of the changes in indicators of the NO metabolism and hemostasis. The obtained results suggest that the registered manifestation of the pro-coagulation and thrombogenic action of succinamides applied in the sub-toxic doses is partially determined by a drop of the vasoactive NO pool that in turn, occurs due to a decline of the NOS activity.

Tick-borne neuroinfections: Clinical characteristics, immunopathogenesis, and new pharmacotherapeutic strategies

To study the semiotics of neurological lesions in patients with tick-borne encephalitis, Ixodes tick-borne borreliosis (ITBB) and mixed infection (MI), their immunopathogenesis, and the possibilities of current pathogenetic pharmacological correction. A total of 220 patients with tick-borne encephalitis, ITBB, and MI concurrent with the syndromes of central nervous system lesions were examined. The immunological studies encompassed the examination of mononuclear cells in the cerebrospinal fluid (CSF), the population and subpopulation composition of lymphocytes, and nitroxidergic processes in the serum and CSF from the total level of final stable nitric oxide metabolites. For pharmacotherapeutic correction, the metabolic drug cytoflavin was used as newly indicated. Cytofluorometric analysis of the CSF cellular composition showed the mononuclear cell predominance of CD3+ (58.6%), CD4+ (57.2%), CD8+ (16.8%) lymphocytes and monocytes (34.4%), which expressed the phenotypic marker CD14+. This reflects the nature of a local immune response: an increase in the immunoregulatory index CD4+/CD8+ from 3.4 to 5.6, respectively, while the normal proportion of these cells in the blood ranges from 1.5 to 2.2. CSF lymphocytes were found to be ready for Fas-mediated apoptosis dependent on the receptor (CD95+ was 64.3%).There was a correlation using the pair correlation coefficient between the total concentration of the metabolites of the nitroxide molecule and the percentage of CD14+ (r=0.5; p<0.05). The paired Wilcoxon test was used to analyze serum NO2, NO3, and NOx, which revealed significant differences in nitrites [2.70 (1.90, 2.95; p=0.001)] and total NO metabolites [18.00 (18.00, 22.60; p=0.006)] and statistically significant changes in nitrates [13.29 (15.70. 20.30; p=0.075)] in patients receiving cytoflavin infusions. The immune response of Th-1 forms between the CSF phagocytic, antigen-presenting, and immunocompetent lymphocytes in patients with tick-borne neuroinfections. The use of cytoflavin as an agent for neurotransmitter support to correct nitroxidergic processes is pathogenetically justified.

PS 08-81 ENDOTHELIAL MARKERS IN HYPERTENSIVES WITH SUBCLINICAL ORGAN DAMAGE

Objective: To investigate stable nitric oxide metabolites (NOx), endothelin-1(ET-1), homocysteine (HC), von Willebrand factor (vWF) levels in patients with essential hypertension (EH) and subclinical organ damage (SOD). Design and Method: We included124 patients (45 men and 79 women) with untreated EH (mean age 51.4 ± 6y, mean EH duration 8.5 ± 7y) and 25 healthy volunteers (10 men and 15 women) with comparable age (47.2 ± 7y). Plasma NOx level was measured by spectrophotometry, ET-1, HC, vWF levels were studied with the help of immunoenzyme assay. Results were processed with Statistica 8.0 software. Results: Microalbuminuria (MAU) was detected in 22% of hypertensives, left ventricular hypertrophy (LVH) in 37% of them. NOx, ET-1, HC, vWF levels were significantly higher in hypertensives (43,2 ± 21 μmol/l, 1,2 ± 0,9 fmol/l, 17,9 ± 7,8 μmol/l, 1,3 ± 0,6 mg/dl, accordingly) than in controls (28,3 ± 9 μmol/l, 0,6 ± 0,5 fmol/l, 10,7 ± 5,8 μmol/l, 1,01 ± 0,7 mg/dl, accordingly) (р < 0,05). The concentration of NOx and ET-1were significantly higher in patients with MAU in comparison to others (46,5 ± 22, 31,3 ± 12 μmol/l and 1,3 ± 0,8, 0,5 ± 0,6 fmol/l, accordingly, р < 0,05). NOx, ET-1 levels were correlated with the presence of MAU (r = 0,48, р = 0,007; r = 0,37 р = 0,03, accordingly). HC, vWF levels did not differ between hypertensives with MAU and normoalbuminuria (р > 0,05). In patients with LVH NOx, ET-1, HC, vWF levels were significantly higher (49,9 ± 19 μmol/l, 1,3 ± 1,1 fmol/l, 20,1 ± 7,6 μmol/l, 1,5 ± 0,7 mg/dl, accordingly) than in those without it (39,8 ± 18 μmol/l, 0,8 ± 0,9 fmol/l, 16,5 ± 7,1 μmol/l, 1,2 ± 0,7 mg/dl accordingly), (р < 0,05). NOx concentration correlated with left ventricular mass index (r = 0,44 р = 0,005), interventricular septum thickness (r = 0,36 р = 0,02) and left ventricle posterior wall thickness (r = 0,44 р = 0,006). The vWF level was correlated with the presence of LVH (r = 0,45, р < 0,009). Conclusions: NOx, ET-1, HC, vWF were elevated in hypertensives with LVH, and NOx, ET-1 levels were higher in hypertensives with MAU. The levels of studied endothelial markers were directly correlated with presence of LVH and MAU

Stable Nitric Oxide Metabolites Levels in Hypertensive Patients With Obesity and High Leptin Level

to investigate levels of the stable nitric oxide metabolites (NOx) in hypertensive patients with obesity and hyperleptinemia. We examined 124 untreated patients (45 men and 79 women) with essential hypertension (EH) (mean age 51.4+/-6.5 years, mean hypertension duration 8.5+/-7.6 years, 64% with visceral obesity) and 25 healthy volunteers (10 men and 15 women with comparable age). NOx levels were significantly higher in hypertensives (43.18+/-21 mol/l) then in controls (28.3+/-9.6 mol/l; p=0.01). Obese hypertensives had lower NOx concentration than nonobese hypertensives (38.8+/-17.9 and 48.5+/-24.7 mol/l, respectively; <0.05). With increase of degree of obesity NOx levels decreased significantly (r=-0.3; p<0.05). NOx levels correlated with presence (r=-0.44; p<0,05) and degree of obesity (r=-0.3; p<0.05). Hypertensives with high leptin levels had lower NOx concentration (29+/-12.2 mol/l), compared to hypertensives with normal leptin levels (34. 7+/-10.5 mol/l) (p<0.05).