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FAST, SPECIFIC AND SENSITIVE DETERMINATION OF S-NITROSOGLUTATHIONE IN BIOLOGICAL SAMPLES - A NOVEL ANALYTICAL PROCEDURE
Publication Only 9
TURBULENT BLOOD FLOW PLAYS AN ESSENTIAL LOCALIZING ROLE IN THE DEVELOPMENT OF ATHEROSCLEROTIC PLAQUES IN EXPERIMENTALLY-INDUCED HYPERCHOLESTEROLEMIA IN RATS
C.M. Prado, S.G. Ramos, M.A. Rossi. Faculty of Medicine of Ribeirao Preto, Department of Pathology, Laboratory of Cell and Molecular Cardiology, Ribeirao Preto, Sao Paulo, Brazil Background: Considering atherosclerosis a focal disease and high levels of plasma cholesterol are closely correlated with its pathogenesis, it is a challenge to explain how equal concentrations of cholesterol bathing the endothelium can produce local rather than global effects on arteries. The focal distribution of atherosclerotic lesions has been considered to be dependent, at least in part, on hydrodynamic factors. The present study was carried out to further test the hypothesis that these forces are an important localizing factor in the pathogenesis of atherosclerosis. Methods: Male young rats feeding a hypercholesterolemic diet were submitted to infra-diaphragmatic aortic constriction for 28days. Results/Conclusions: These animals develop a normotensive prestenotic region with laminar blood flow that serves as control for a normotensive poststenotic region with turbulent blood flow. Our findings demonstrated that the combination of turbulent blood flow and low wall shear stress in the presence of hypercholesterolemia and oxidative stress creates conditions to the formation of focally distributed incipient atherosclerotic plaques observed in the poststenotic segment. In contrast, only diffuse fatty streaks could be observed in the normotensive prestenotic segment with laminar blood flow and normal wall shear stress in the presence of hypercholesterolemia and oxidative stress. Although hemodynamic forces are not by themselves responsible for the pathogenesis of atherosclerosis, they prime the local vascular wall in which the lesion develop. Further studies are required to establish how hemodynamic forces are detected and transduced into chemical signaling by the cells of the artery wall and then converted into pathophysiologically relevant phenotypic changes. 10
ASSOCIATIONS BETWEEN WAIST CIRCUMFERENCE AND OTHER CARDIOVASCULAR RISK FARCTORS
A. Kalvelis, G. Bahs, A. Lejnieks, I. Stukena. Department of Internal Diseases, Riga Stradins University, Riga, Latvia
EFFECT OF SEVELAMER HYDROCLORIDE ON SERUM LIPIDS AND LIPOPROTEIN A IN HEMODIALISYS PATIENTS
J.A. Díaz-Peromingo 1 , P. Pesqueira-Fontán 1 , D. Güimil-Carbajal 2 , A. Albán-Salgado 3 , S. Molinos-Castro 1 , M. Pena-Seijo 4 , J. Sánchez-Leira 1 , F. García-Suárez 1 , J. Saborido-Froján 1 , E. Padín-Paz 1 , M. Iglesias-Gallego 1 , J. Naveiro-Soneira 1 , M. Ferreira 5 . 1 Internal Medicine, Hospital Da Barbanza, Riveira, Spain; 2 Nephrology, Hospital Da Barbanza, Riveira, Spain; 3 Laboratory Unit, Hospital Da Barbanza, Riveira, Spain; 4 Internal Medicine, Hospital De Conxo, Santiago, Spain; 5 Internal Medicine, Centro Hospitalar Alto Minho, Viana Do Castelo, Portugal Introduction: Sevelamer hydrochloride, a new compound non-aluminum and non-calcium phosphate binder, has been introduced in haemodialysis patients. Besides the effect on phosphate, it has been associated with reduction of coronary and aortic calcification and effects on lipid metabolism. Our aim is to study the effect of sevelamer on lipid measures especially Lp(a). Material and Methods: We studied two groups. Group 1: patients without treatment with sevelamer. Group 2: patients with sevelamer. Sex, age, total cholesterol, triglycerides, LDL, HDL, VLDL, total cholesterol/HDL index, LDL/HDL index and Lp(a) were reported. Statistical analysis was made using descriptive statistics and a T test to compare means when needed. Results: Group 1: 20 patients (7 men and 13 women). Mean age 65.60 years. Group 2: 24 patientes (10 men and 14 women). Mean age 63.88 years. Results were as follows: total cholesterol 172.95 versus 143.88 mg/dl (p=0.021*, SS), triglycerides 139.75 versus 133.46 mg/dl (p=0.418, NS), HDL 41.35 versus 25.13 mg/dl (p=0.025*, SS), Lp(a) 4.70 versus 3.92 (p=0.049*, SS), VLDL 13.40 versus 18.88 mg/dl (p=0.231, NS), LDL 113.45 versus 51.96 mg/dl (p=0.000*, SS), Col/HDL 4.70 versus 4.47 (p=0.123, NS), LDL/HDL 3.17 versus 2.44 (p=0.079, NS). Conclusions: 1. Sevelamer hydrochloride improves significantly total cholesterol and LDL in patients on hemodialisys. 2. Lp (a) concentration also improves significantly in patients taking sevelamer as compared with those without this treatment. 3. In our study, HDL concentrations decreases significantly after the addition of sevelamer to current treatment. 12
FAST, SPECIFIC AND SENSITIVE DETERMINATION OF S-NITROSOGLUTATHIONE IN BIOLOGICAL SAMPLES - A NOVEL ANALYTICAL PROCEDURE
E. Bramanti 1 , C. Vecoli 1 , A. Pompella 2 , R. Barsacchi 3 , R. Baldassini 1 , D. Neglia 3 , M. Franzini 2 , A. Paolicchi 2 . 1 National Research Council-CNR, Inst. for Chemico-Physical Processes, Pisa, Italy; 2 Dept. of Experimental Pathology, University of Pisa Medical School, Pisa, Italy; 3 National Research Council-CNR, Inst. of Clinical Physiology, Pisa, Italy It has been recently hypothesized that nitric oxide (NO) can have an endocrine function, exerting a control over vascular tone at a distance from sites of biosynthesis/administration. As NO is a very reactive molecule, such effects should be mediated through more stable, bioactive species. Such a role may be played by covalently bound sulfhydryl-NO low mol. weight adducts, i.e., S-nitroso-glutathione (GSNO) and other S-nitrosothiols. However, research in this field has long been hindered by a lack of adequate methodology for collection of data. Controversy still exists even over the level of S-nitrosothiols in plasma, with differences of even 3–4 orders of magnitude among the reported values. The Authors have recently patented a novel analytical procedure for the determination of GSNO in biological samples (Italian Pat. Appl. No. PI/2006/A/000093), which is rapid, reproducible and considerably more sensitive than previous assays. It is based on the enzymatic decomposition of GSNO by gamma-glutamyltransferase (GGT) and the fast decomposition of its product CysGlyNO (CGNO) by copper. The developed NO is detected by the reaction with 4,5-diaminofluorescein (DAF-2) and spectrofluorimetric detection of the triazole derivative. The limit of quantitation (LOQc) of the proposed method is 20 nM with a CV of 5.5% at 0.3 mM concentration level and a 20-300 nmol/L linear dynamic range depending on DAF-2 concentration. Thus, detection limits two orders of magnitude lower are achieved as compared to direct UV detection techniques, making the assay suitable to investigate formation, metabolism, and pathophysiology of GSNO in clinical conditions.
77th Congress of the European Atherosclerosis Society, April 26–29, 2008, Istanbul, Turkey
PUBLICATION ONLY
Aim: Aim of the study is to detect correlation of abdominal obesity- us significant risk factor (RF) of cardio-vascular diseases with other RF in out patients. Methods: We have assessed correlation of waist circumference (WC) with sex, age, BMI, TC, HDL-C, non-HDL-C, CRP, systolic and diastolic blood pressure (SBP, DBP) in 1094 outpatients by dividing patients into 2 subgroups: A group included men having WC≤94 cm and women having WC≤80cm, and B group - men having WC≥95cm and women having WC≥81cm. Results: Analysis of all the patients revealed significant correlation of WC only with age, BMI, height, SBP, DBP, TC, non-HDL-C and CRP. WC in women does not correlate with height, but in men WC tends to correlate with CRP (p=0.055). Men of both groups had higher levels of SBP and DBP, but lower values of HDL-C, compared to women. In group B there were significantly (p<0.001) higher values of SBP, DBP, TC, non-HDL-C and CRP, but lower–of HDL-C. TC levels were similar in both men and women of group A, while in group B TC levels were higher in women. CRP levels were higher in men of the group A, but similar in men and women of group B. Conclusion: Our results revealed that patients having larger WC have higher levels of SBP, DBP, TC, non-HDL-C and CRP, but lower–HDL-C levels. Men had higher SBP, DBP and non-HDL-C. Increased WC significantly aggravates effects of other CVD risk factors, and such a risk is more evident in women.
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TURBULENT BLOOD FLOW PLAYS AN ESSENTIAL LOCALIZING ROLE IN THE DEVELOPMENT OF ATHEROSCLEROTIC PLAQUES IN EXPERIMENTALLY-INDUCED HYPERCHOLESTEROLEMIA IN RATS
C.M. Prado, S.G. Ramos, M.A. Rossi. Faculty of Medicine of Ribeirao Preto, Department of Pathology, Laboratory of Cell and Molecular Cardiology, Ribeirao Preto, Sao Paulo, Brazil Background: Considering atherosclerosis a focal disease and high levels of plasma cholesterol are closely correlated with its pathogenesis, it is a challenge to explain how equal concentrations of cholesterol bathing the endothelium can produce local rather than global effects on arteries. The focal distribution of atherosclerotic lesions has been considered to be dependent, at least in part, on hydrodynamic factors. The present study was carried out to further test the hypothesis that these forces are an important localizing factor in the pathogenesis of atherosclerosis. Methods: Male young rats feeding a hypercholesterolemic diet were submitted to infra-diaphragmatic aortic constriction for 28days. Results/Conclusions: These animals develop a normotensive prestenotic region with laminar blood flow that serves as control for a normotensive poststenotic region with turbulent blood flow. Our findings demonstrated that the combination of turbulent blood flow and low wall shear stress in the presence of hypercholesterolemia and oxidative stress creates conditions to the formation of focally distributed incipient atherosclerotic plaques observed in the poststenotic segment. In contrast, only diffuse fatty streaks could be observed in the normotensive prestenotic segment with laminar blood flow and normal wall shear stress in the presence of hypercholesterolemia and oxidative stress. Although hemodynamic forces are not by themselves responsible for the pathogenesis of atherosclerosis, they prime the local vascular wall in which the lesion develop. Further studies are required to establish how hemodynamic forces are detected and transduced into chemical signaling by the cells of the artery wall and then converted into pathophysiologically relevant phenotypic changes. 10
ASSOCIATIONS BETWEEN WAIST CIRCUMFERENCE AND OTHER CARDIOVASCULAR RISK FARCTORS
A. Kalvelis, G. Bahs, A. Lejnieks, I. Stukena. Department of Internal Diseases, Riga Stradins University, Riga, Latvia
EFFECT OF SEVELAMER HYDROCLORIDE ON SERUM LIPIDS AND LIPOPROTEIN A IN HEMODIALISYS PATIENTS
J.A. Díaz-Peromingo 1 , P. Pesqueira-Fontán 1 , D. Güimil-Carbajal 2 , A. Albán-Salgado 3 , S. Molinos-Castro 1 , M. Pena-Seijo 4 , J. Sánchez-Leira 1 , F. García-Suárez 1 , J. Saborido-Froján 1 , E. Padín-Paz 1 , M. Iglesias-Gallego 1 , J. Naveiro-Soneira 1 , M. Ferreira 5 . 1 Internal Medicine, Hospital Da Barbanza, Riveira, Spain; 2 Nephrology, Hospital Da Barbanza, Riveira, Spain; 3 Laboratory Unit, Hospital Da Barbanza, Riveira, Spain; 4 Internal Medicine, Hospital De Conxo, Santiago, Spain; 5 Internal Medicine, Centro Hospitalar Alto Minho, Viana Do Castelo, Portugal Introduction: Sevelamer hydrochloride, a new compound non-aluminum and non-calcium phosphate binder, has been introduced in haemodialysis patients. Besides the effect on phosphate, it has been associated with reduction of coronary and aortic calcification and effects on lipid metabolism. Our aim is to study the effect of sevelamer on lipid measures especially Lp(a). Material and Methods: We studied two groups. Group 1: patients without treatment with sevelamer. Group 2: patients with sevelamer. Sex, age, total cholesterol, triglycerides, LDL, HDL, VLDL, total cholesterol/HDL index, LDL/HDL index and Lp(a) were reported. Statistical analysis was made using descriptive statistics and a T test to compare means when needed. Results: Group 1: 20 patients (7 men and 13 women). Mean age 65.60 years. Group 2: 24 patientes (10 men and 14 women). Mean age 63.88 years. Results were as follows: total cholesterol 172.95 versus 143.88 mg/dl (p=0.021*, SS), triglycerides 139.75 versus 133.46 mg/dl (p=0.418, NS), HDL 41.35 versus 25.13 mg/dl (p=0.025*, SS), Lp(a) 4.70 versus 3.92 (p=0.049*, SS), VLDL 13.40 versus 18.88 mg/dl (p=0.231, NS), LDL 113.45 versus 51.96 mg/dl (p=0.000*, SS), Col/HDL 4.70 versus 4.47 (p=0.123, NS), LDL/HDL 3.17 versus 2.44 (p=0.079, NS). Conclusions: 1. Sevelamer hydrochloride improves significantly total cholesterol and LDL in patients on hemodialisys. 2. Lp (a) concentration also improves significantly in patients taking sevelamer as compared with those without this treatment. 3. In our study, HDL concentrations decreases significantly after the addition of sevelamer to current treatment. 12
FAST, SPECIFIC AND SENSITIVE DETERMINATION OF S-NITROSOGLUTATHIONE IN BIOLOGICAL SAMPLES - A NOVEL ANALYTICAL PROCEDURE
E. Bramanti 1 , C. Vecoli 1 , A. Pompella 2 , R. Barsacchi 3 , R. Baldassini 1 , D. Neglia 3 , M. Franzini 2 , A. Paolicchi 2 . 1 National Research Council-CNR, Inst. for Chemico-Physical Processes, Pisa, Italy; 2 Dept. of Experimental Pathology, University of Pisa Medical School, Pisa, Italy; 3 National Research Council-CNR, Inst. of Clinical Physiology, Pisa, Italy It has been recently hypothesized that nitric oxide (NO) can have an endocrine function, exerting a control over vascular tone at a distance from sites of biosynthesis/administration. As NO is a very reactive molecule, such effects should be mediated through more stable, bioactive species. Such a role may be played by covalently bound sulfhydryl-NO low mol. weight adducts, i.e., S-nitroso-glutathione (GSNO) and other S-nitrosothiols. However, research in this field has long been hindered by a lack of adequate methodology for collection of data. Controversy still exists even over the level of S-nitrosothiols in plasma, with differences of even 3–4 orders of magnitude among the reported values. The Authors have recently patented a novel analytical procedure for the determination of GSNO in biological samples (Italian Pat. Appl. No. PI/2006/A/000093), which is rapid, reproducible and considerably more sensitive than previous assays. It is based on the enzymatic decomposition of GSNO by gamma-glutamyltransferase (GGT) and the fast decomposition of its product CysGlyNO (CGNO) by copper. The developed NO is detected by the reaction with 4,5-diaminofluorescein (DAF-2) and spectrofluorimetric detection of the triazole derivative. The limit of quantitation (LOQc) of the proposed method is 20 nM with a CV of 5.5% at 0.3 mM concentration level and a 20-300 nmol/L linear dynamic range depending on DAF-2 concentration. Thus, detection limits two orders of magnitude lower are achieved as compared to direct UV detection techniques, making the assay suitable to investigate formation, metabolism, and pathophysiology of GSNO in clinical conditions.
77th Congress of the European Atherosclerosis Society, April 26–29, 2008, Istanbul, Turkey
PUBLICATION ONLY
Aim: Aim of the study is to detect correlation of abdominal obesity- us significant risk factor (RF) of cardio-vascular diseases with other RF in out patients. Methods: We have assessed correlation of waist circumference (WC) with sex, age, BMI, TC, HDL-C, non-HDL-C, CRP, systolic and diastolic blood pressure (SBP, DBP) in 1094 outpatients by dividing patients into 2 subgroups: A group included men having WC≤94 cm and women having WC≤80cm, and B group - men having WC≥95cm and women having WC≥81cm. Results: Analysis of all the patients revealed significant correlation of WC only with age, BMI, height, SBP, DBP, TC, non-HDL-C and CRP. WC in women does not correlate with height, but in men WC tends to correlate with CRP (p=0.055). Men of both groups had higher levels of SBP and DBP, but lower values of HDL-C, compared to women. In group B there were significantly (p<0.001) higher values of SBP, DBP, TC, non-HDL-C and CRP, but lower–of HDL-C. TC levels were similar in both men and women of group A, while in group B TC levels were higher in women. CRP levels were higher in men of the group A, but similar in men and women of group B. Conclusion: Our results revealed that patients having larger WC have higher levels of SBP, DBP, TC, non-HDL-C and CRP, but lower–HDL-C levels. Men had higher SBP, DBP and non-HDL-C. Increased WC significantly aggravates effects of other CVD risk factors, and such a risk is more evident in women.
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