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MTHFR Polymorphism in patients with stroke and deep venous thrombosis
94
Crouse
cholesterol, sialic acid, CRP and apo B were significantly greater in patients with AS than controls (table 1). Table I
Total chol0mnolfl) TG 0mnol/1) tlDL-C 0mnol/1) Apo At (g/l) Apo B (g/l) Lipoprotehl(a)(g/1) CRPm(rag/l) Sialic acid (mg/dl)
AS
Controls
p Value
5.26 (I.I2) 1.7I (0.65) 1.34 (0.32) 1.47 (0.26) 1.04 (0.25) 0.35 (0.42) 3.3 (3.9) 81.0 (it.2)
4.39 (0.94) 1.42 (0.56) 1.42 (0.28) 1.49 (0.39) 0.80 (0A8) 0.23 (0.34) 3A (4.7) 71.0 (I3.I)
0.0025 0.74 0.54 0.83 0.000I 0.24* 0.05I* 0.0it*
group (34.0% and 29.8% in ST and DVT patients, respectively) then in controls (24.8%), however, the difference was not significant. The frequency ofMTHFR TT genotype distribution showed a significant difference between the control and ST group (p-0.004), and DVT group (p-0.001). The frequency of homozygous T allele genotype in ST was 12.5%, in DVT 12.8%, and in control group 0.9%. The odds ratio for ST and DVT patients was 17.571% (CI, 3.553 86.897) and 16.098 (3.095 83.727), respectively, revealing a significant risk for persons with homozygous TT genotype. These preliminary results suggest the potential association between the MTHFR gene polymorphism and development of peripheral vascular disease. In the future, this ongoing study is anticipated to include a larger group of subjects, hopefully leading to more decisive conclusions. F~
*Mmm-WhitneyU test. Multiple regression analysis of the whole data set showed no correlation between effective orifice area and any parameter in either group. These results provide evidence that AS is an atherosclerotic disease in showing dependence both on lipoproteins and on inflammatory markers. ~ 1 - ~ MEASURING EFFECTS ON INTIMA MEDIA T H I C K N E S S : AN EVALUATION OF ROSUVASTATIN THE M E T E O R STUDY J.R. Crouse 1, D.E. Grobbee2, D.H. O' Lea@, J.J.P. Kastelein4, M.L. Bots 2, .__{5. 1 Wake Forest University G.W. Evans 1, M.K. Palmer5, J.S. Raichlen
Baptist Medical Center, Winston-Salem, NC, USA," 2University Medical Center Utrecht, The Netherlands," 3New England Medical Center, Boston, MA, USA," 4Academic Medical Center, Amsterdam, The Netherlands," 5AstraZeneca, Macclesfield, Cheshire, UK," 6AstraZeneca, Wayne, PA, USA Ultrasound assessment of carotid artery intima media thickness (IMT) is a valuable surrogate marker of atherosclerosis and cardiovascular disease. Lipid lowering with HMG-CoA reductase inhibitors (statins) retards progression or, at high doses, may promote regression of carotid IMT in subjects with a high risk of coronary heart disease (CHD). The objective of this randomized, double-blind, placebo-controlled, multinational study (4522IL/0081) is to evaluate the effect of the efficacious statin, rosuvastatin (Crestor) on carotid IMT in asymptomatic subjects with a low risk of CHD events. 840 asymptomatic subjects with evidence of atherosclerotic disease by a thickened carotid IMT (maximum IMT )1.2 and <3.5 nun) will be randomized (5:2) to 104 weeks of treatment with rosuvastatin (40 mg/day) or placebo to obtain a minimum evaluable study population of 581. The primary efficacy end point will be the change from baseline to end of treatment in the mean maximum IMT (MeanMax IMT) determined from the 12 maximum IMT measurements of the near and far walls of the right and left common carotid artery (CCA), carotid bulb and internal carotid artery (ICA). Secondary efficacy end points will include the change from baseline to end of treatment in: MeanMax IMT of each of the CCA, carotid bulb and ICA; mean IMT of the CCA; lipid parameters; and inflammatory markers. A safety evaluation will also be conducted. A positive outcome in this study would support statin treatment in subjects with a low risk of CHD events and evidence of atherosclerotic disease by a thickened carotid IMT. ~ 1 - ~ M T H F R POLYMORPHISM IN PATIENTS W I T H STROKE AND DEEP VENOUS T H R O M B O S I S M. Cubrilo-Turek 1, E. Topic 2, A. Begonja2, M. Stefanovic 2, A.-M. Simundic 2, V. Demarin3. JSveti Duh General Hospital, 2Clinical
Institute of Chemistry, 3Department of Neurology, School of Medicine, University of Zagreb and Sestre Milosrdnice, Zagreb, Croatia Hyperhomocysteinemia is an independent risk factor for vascular disease, in which besides nutritional, genetic factors are also involved. The polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene encoding for the enzyme, that controls homocysteine metabolism, predisposed to hyperchomocysteinemia, suggesting that MTHFR TT genotype is associated with an increased risk of vascular disease. The aim of the study was to evaluate the MTHFR polymorphism in 56 patients with stroke (ST) and 47 patients with deep venous thrombosis (DVT), and to compare it with control group (n-111). Polymorhic MTHFR alleles were detected by DNA isolated from EDTA blood by PCR-RFLP and RE Hinf I (G-AnTC). Quality controls containing known genotypes were included in each run. The results showed higher MTHFR T allele frequencies in either patient
EFFECTS OF THIAZOLIDINEDIONES ON PROLIFERATION AND PROTEOGLYCAN BIOSYNTHESIS IN HUMAN VASCULAR SMOOTH MUSCLE CELLS
S.T. de Dios 1.3, G.L.R. Jennings2'3, RJ. Little 1.3. J Cell Biology of Diabetes Laboratory, Baker Medical Research Institute," 2Baker Medical Research Institute," 3Monash University, Melbourne, Victoria, Australia Diabetes is associated with accelerated development of vascular disease, at least partially due to the pathological effects of hyperglycemia. A major therapeutic option is vascular reconstruction by coronary artery by-pass grafting employing internal mammary arteries (IMA), radial arteries (RA) and saphenous veins (SV). Thiazolidinediones (TZDs) are the newest agents for the treatment of hyperglycemia in Type 2 diabetes and we are interested in the direct vascular actions of TZDs on atherogenic properties of these three vessels. We investigated the effects of troglitazone, rosiglitazone and pioglitazone on proliferation and proteoglycan synthesis in human vascular smooth muscle cells (VSMCs). Proliferation was assessed in VSMCs derived from IMA, RA and SV by cell counting. Proteoglycan production in IMAs was investigated through 35S-sulphate incorporation and SDS-PAGE sizing (4-15%). All three TZDs inhibited proliferation in the three cell types with an order of potency troglitazone> pioglitazone-rosiglitazone. Maximum inhibition was approximately 50% and troglitazone showed half-maximal inhibition activity between 1 and 3~tM. Rosiglitazone and pioglitazone only showed inhibition at very high concentrations (30 and 100~tM). All three compounds inhibited basal proteoglycan synthesis, while SDS-PAGE showed slight reductions in size. The TZDs also inhibited proteoglycan synthesis stimulated by the atherogenic growth factors PDGF-BB and TGF[31. The reduction in proteoglycan size indicates structural modification and potentially reduced binding ability to atherogenic lipoproteins and inhibition of proliferation may also reduce potentially atherosclerotic vascular changes. These data suggest that direct vascular effects may contribute to the antiatherosclerotic actions of TZDs. F 3 - ~ PATHOPHYSIOLOGICAL ROLE OF LIPID PEROXIDATION IN PATIENTS W I T H ACUTE AND CHRONIC CORONARY SYNDROMES E. De Fanti 1, M. Rugolotto 2, A. Fasolin 1, E. Visentin2, M. Barbiero 2, G. Rigatelli 2, R. Schiavon 1. JLaboratory Medicine, 2Cardiovascular
Department, Legnago Hospital, Legnago (Verona), Italy Introduction: Accumulating evidence supports the role of lipid peroxidation
in inflammatory processes and atherosclerosis. Aim: To measure the levels of plasma malondialdehyde (MDA), an index of lipid peroxidation, in patients (pts) with acute and chronic coronary syndromes (ACS, CCS) and to evaluate its possible role in inducing endothelial disfunction, as assessed by s-VCAM-1 and s-ICAM-1, and inflammation, by C-reactive protein (CRP). Methods: We obtained plasma samples from 76 pts. 27 pts had ACS (14 unstable angina, 13 AMI). 37 pts had CCS (stable angina). 12 were control non-coronary subjects. MDA was assayed by a spectrophotometric method; s-VICAM-1 and s-ICAM-1 by ELISA (Biosource International); CRP by immunonephelometry (Immage, Beckman). Results:
ACS CCS Controls
MDA(tnnol/L)
sVCAM-I(ug/L) slCAM-I(ug/L) CRP (rag/L)
9.[9 (7.34 [0.82)** 9.64 (6.56 [2.42)** 5.87 (3.65~A5)
749 (563~3[)** 389 (360~00) 415 (38IM52)
3[ [ (25[ 370)** 309 (260 352)** 204 (I88 230)
[0.2 (5.7 [7A)* 4A (2.5 5.0) 3A (0.5 5.2)
Values:median(hlterquartilerange). (**):p<.0i vs conlrols;(*): p<.05 vs controls.
73rd EAS Congress
Crouse
cholesterol, sialic acid, CRP and apo B were significantly greater in patients with AS than controls (table 1). Table I
Total chol0mnolfl) TG 0mnol/1) tlDL-C 0mnol/1) Apo At (g/l) Apo B (g/l) Lipoprotehl(a)(g/1) CRPm(rag/l) Sialic acid (mg/dl)
AS
Controls
p Value
5.26 (I.I2) 1.7I (0.65) 1.34 (0.32) 1.47 (0.26) 1.04 (0.25) 0.35 (0.42) 3.3 (3.9) 81.0 (it.2)
4.39 (0.94) 1.42 (0.56) 1.42 (0.28) 1.49 (0.39) 0.80 (0A8) 0.23 (0.34) 3A (4.7) 71.0 (I3.I)
0.0025 0.74 0.54 0.83 0.000I 0.24* 0.05I* 0.0it*
group (34.0% and 29.8% in ST and DVT patients, respectively) then in controls (24.8%), however, the difference was not significant. The frequency ofMTHFR TT genotype distribution showed a significant difference between the control and ST group (p-0.004), and DVT group (p-0.001). The frequency of homozygous T allele genotype in ST was 12.5%, in DVT 12.8%, and in control group 0.9%. The odds ratio for ST and DVT patients was 17.571% (CI, 3.553 86.897) and 16.098 (3.095 83.727), respectively, revealing a significant risk for persons with homozygous TT genotype. These preliminary results suggest the potential association between the MTHFR gene polymorphism and development of peripheral vascular disease. In the future, this ongoing study is anticipated to include a larger group of subjects, hopefully leading to more decisive conclusions. F~
*Mmm-WhitneyU test. Multiple regression analysis of the whole data set showed no correlation between effective orifice area and any parameter in either group. These results provide evidence that AS is an atherosclerotic disease in showing dependence both on lipoproteins and on inflammatory markers. ~ 1 - ~ MEASURING EFFECTS ON INTIMA MEDIA T H I C K N E S S : AN EVALUATION OF ROSUVASTATIN THE M E T E O R STUDY J.R. Crouse 1, D.E. Grobbee2, D.H. O' Lea@, J.J.P. Kastelein4, M.L. Bots 2, .__{5. 1 Wake Forest University G.W. Evans 1, M.K. Palmer5, J.S. Raichlen
Baptist Medical Center, Winston-Salem, NC, USA," 2University Medical Center Utrecht, The Netherlands," 3New England Medical Center, Boston, MA, USA," 4Academic Medical Center, Amsterdam, The Netherlands," 5AstraZeneca, Macclesfield, Cheshire, UK," 6AstraZeneca, Wayne, PA, USA Ultrasound assessment of carotid artery intima media thickness (IMT) is a valuable surrogate marker of atherosclerosis and cardiovascular disease. Lipid lowering with HMG-CoA reductase inhibitors (statins) retards progression or, at high doses, may promote regression of carotid IMT in subjects with a high risk of coronary heart disease (CHD). The objective of this randomized, double-blind, placebo-controlled, multinational study (4522IL/0081) is to evaluate the effect of the efficacious statin, rosuvastatin (Crestor) on carotid IMT in asymptomatic subjects with a low risk of CHD events. 840 asymptomatic subjects with evidence of atherosclerotic disease by a thickened carotid IMT (maximum IMT )1.2 and <3.5 nun) will be randomized (5:2) to 104 weeks of treatment with rosuvastatin (40 mg/day) or placebo to obtain a minimum evaluable study population of 581. The primary efficacy end point will be the change from baseline to end of treatment in the mean maximum IMT (MeanMax IMT) determined from the 12 maximum IMT measurements of the near and far walls of the right and left common carotid artery (CCA), carotid bulb and internal carotid artery (ICA). Secondary efficacy end points will include the change from baseline to end of treatment in: MeanMax IMT of each of the CCA, carotid bulb and ICA; mean IMT of the CCA; lipid parameters; and inflammatory markers. A safety evaluation will also be conducted. A positive outcome in this study would support statin treatment in subjects with a low risk of CHD events and evidence of atherosclerotic disease by a thickened carotid IMT. ~ 1 - ~ M T H F R POLYMORPHISM IN PATIENTS W I T H STROKE AND DEEP VENOUS T H R O M B O S I S M. Cubrilo-Turek 1, E. Topic 2, A. Begonja2, M. Stefanovic 2, A.-M. Simundic 2, V. Demarin3. JSveti Duh General Hospital, 2Clinical
Institute of Chemistry, 3Department of Neurology, School of Medicine, University of Zagreb and Sestre Milosrdnice, Zagreb, Croatia Hyperhomocysteinemia is an independent risk factor for vascular disease, in which besides nutritional, genetic factors are also involved. The polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene encoding for the enzyme, that controls homocysteine metabolism, predisposed to hyperchomocysteinemia, suggesting that MTHFR TT genotype is associated with an increased risk of vascular disease. The aim of the study was to evaluate the MTHFR polymorphism in 56 patients with stroke (ST) and 47 patients with deep venous thrombosis (DVT), and to compare it with control group (n-111). Polymorhic MTHFR alleles were detected by DNA isolated from EDTA blood by PCR-RFLP and RE Hinf I (G-AnTC). Quality controls containing known genotypes were included in each run. The results showed higher MTHFR T allele frequencies in either patient
EFFECTS OF THIAZOLIDINEDIONES ON PROLIFERATION AND PROTEOGLYCAN BIOSYNTHESIS IN HUMAN VASCULAR SMOOTH MUSCLE CELLS
S.T. de Dios 1.3, G.L.R. Jennings2'3, RJ. Little 1.3. J Cell Biology of Diabetes Laboratory, Baker Medical Research Institute," 2Baker Medical Research Institute," 3Monash University, Melbourne, Victoria, Australia Diabetes is associated with accelerated development of vascular disease, at least partially due to the pathological effects of hyperglycemia. A major therapeutic option is vascular reconstruction by coronary artery by-pass grafting employing internal mammary arteries (IMA), radial arteries (RA) and saphenous veins (SV). Thiazolidinediones (TZDs) are the newest agents for the treatment of hyperglycemia in Type 2 diabetes and we are interested in the direct vascular actions of TZDs on atherogenic properties of these three vessels. We investigated the effects of troglitazone, rosiglitazone and pioglitazone on proliferation and proteoglycan synthesis in human vascular smooth muscle cells (VSMCs). Proliferation was assessed in VSMCs derived from IMA, RA and SV by cell counting. Proteoglycan production in IMAs was investigated through 35S-sulphate incorporation and SDS-PAGE sizing (4-15%). All three TZDs inhibited proliferation in the three cell types with an order of potency troglitazone> pioglitazone-rosiglitazone. Maximum inhibition was approximately 50% and troglitazone showed half-maximal inhibition activity between 1 and 3~tM. Rosiglitazone and pioglitazone only showed inhibition at very high concentrations (30 and 100~tM). All three compounds inhibited basal proteoglycan synthesis, while SDS-PAGE showed slight reductions in size. The TZDs also inhibited proteoglycan synthesis stimulated by the atherogenic growth factors PDGF-BB and TGF[31. The reduction in proteoglycan size indicates structural modification and potentially reduced binding ability to atherogenic lipoproteins and inhibition of proliferation may also reduce potentially atherosclerotic vascular changes. These data suggest that direct vascular effects may contribute to the antiatherosclerotic actions of TZDs. F 3 - ~ PATHOPHYSIOLOGICAL ROLE OF LIPID PEROXIDATION IN PATIENTS W I T H ACUTE AND CHRONIC CORONARY SYNDROMES E. De Fanti 1, M. Rugolotto 2, A. Fasolin 1, E. Visentin2, M. Barbiero 2, G. Rigatelli 2, R. Schiavon 1. JLaboratory Medicine, 2Cardiovascular
Department, Legnago Hospital, Legnago (Verona), Italy Introduction: Accumulating evidence supports the role of lipid peroxidation
in inflammatory processes and atherosclerosis. Aim: To measure the levels of plasma malondialdehyde (MDA), an index of lipid peroxidation, in patients (pts) with acute and chronic coronary syndromes (ACS, CCS) and to evaluate its possible role in inducing endothelial disfunction, as assessed by s-VCAM-1 and s-ICAM-1, and inflammation, by C-reactive protein (CRP). Methods: We obtained plasma samples from 76 pts. 27 pts had ACS (14 unstable angina, 13 AMI). 37 pts had CCS (stable angina). 12 were control non-coronary subjects. MDA was assayed by a spectrophotometric method; s-VICAM-1 and s-ICAM-1 by ELISA (Biosource International); CRP by immunonephelometry (Immage, Beckman). Results:
ACS CCS Controls
MDA(tnnol/L)
sVCAM-I(ug/L) slCAM-I(ug/L) CRP (rag/L)
9.[9 (7.34 [0.82)** 9.64 (6.56 [2.42)** 5.87 (3.65~A5)
749 (563~3[)** 389 (360~00) 415 (38IM52)
3[ [ (25[ 370)** 309 (260 352)** 204 (I88 230)
[0.2 (5.7 [7A)* 4A (2.5 5.0) 3A (0.5 5.2)
Values:median(hlterquartilerange). (**):p<.0i vs conlrols;(*): p<.05 vs controls.
73rd EAS Congress