- Email: [email protected]
M.650 Non-stenotic echolucent plaques in the carotid arteries in clinically healthy, 58-year old men predict cardiovascular events
150
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Miscellaneous
Sasiela EFFECT OF ATORVASTATIN ON LIPOPROTEIN SUBFRACTION PROFILES IN PATIENTS W I T H DYSLIPIDEMIA
W. Sasiela, H. Silbershatz, J. Ma, E. Villagio. Pfizer Inc, New York, USA
Background: A preponderance of small dense LDL is strongly associated with elevated CHD risk. Evidence fi'om small-scale, single-dose studies suggests that additional to reducing overall LDL-C, some statins can qualitatively change LDL subfi'action profile. Methods: NASDAC was a multicenter, double-blind, randomized trial comparing atorvastatin efficacy and safety at starting doses of 10, 20, 40 and 80mg. In a reU'ospective subgroup analysis, atorvastatin effects on various lipoprotein subfi'action profiles were determined using NMR technology. The subgroup population comprised 190 patients who were candidates for lipid-lowering therapy according to NCEP III criteria, had baseline TG >2.3mmol/L and who wele randomized to atorvastatin 10mg (n=40), 20mg (n=55), 40mg (n=47), or 80mg (n=48). Subfi'action analyses were conducted using NMR LipoIh'ofile ®. Results: In the subgroup population,atorvastatin significantly reduced LDL-C levels (measured by standat'd methods) at all doses (P<0.001; 10mg: -34%; 20mg: -40%; 40mg: -49%; 80mg: -52%). Reductions in LDL-C, TC and TG as measured by NMR were similar to those obtained by conventional methods. However, HDL increases were markedly higher when determined by NMR. At all doses, atorvastatin significantly leduced the proportion of LDL pat'ticles that ale small, dense LDL (-15% to -19%, P<0.005), reduced absolute number of LDL pat'ticles (P<0.0001), incleased LDL pat'ticle size (P<0.0001) and improved HDL-subfi'action profile. Conclusion: Atorvastatin beneficially altered the atherogenic lipid profile in dyslipidemia patients. As expected, atorvastatin significantly reduced LDL-C and LDL pat'ticle numbers. Furthermore, atorvastatin significantly decreased LDL pat'ticle density, resulting in a shift fi'om small, dense LDL to more buoyant and less atherogenic particles.
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OXIDIZED LDL IS ASSOCIATED WITH CAROTID IMT M•.649] AND FEATURES OF THE INSULIN RESISTANCE SYNDROME P.G. Scheffer, R.M.A. Henry, E.J.M. Wever, G. Bos, R.J. Heine, J.M. Dekker, C.D.A. Stehouwer, T. Teerlink. Institute for Cardiovascular
Research, VU University Medical Center, Amsterdam, The Netherlands EFFECTS OF PROBUCOL AND PRAVASTATIN IN HYPERCHOLESTEROLEMIC PATIENTS AGED 75 YEARS OR OLDER
Y. Sawayama, N. Maeda, M. Tatsukawa, C. Shimizu, J. Hayashi. Dept. of
General Medicine, Kyushu University Hospital, Fukuoka, Japan The plesent study involved post hoc assessment of the efficacy of lipidlowering therapy in two subsets of the Fukuoka Atherosclerosis Trial (FAST) patients (>75 years old or <75 yeat's old).The FAST cohort of 246 asymptomatic hypercholesterolemic patients (mean age: 66 yeat's) included 76 patients aged >75 years. Patients were randomized to receive probucol at 500 mg/day (n=82) or pravastatin at 10 mg/day (n=83), or were assigned to a control group treated by diet alone (n=81). The change of intima-media thickness (IMT) in the common carotid at'tery was the primary endpoint and the incidence of major cardiovascular events was the secondary endpoint.In patients >75 yeat's old, 2 yeat's of probucol or pravastatin therapy caused a significant decrease of IMT (by 5.8% and 5.5%, respectively; both p<0.05). These lipid-lowering drugs also significantly reduced the IMT in patients <75 yeat's old (by 18.7% and 13.3%, respectively; both p<0.01). In contrast, control patients >75 yeat's old showed a significant increase of IMT compaled with patients <75 yeat's old (by 34.3% and 12.1%, respectively; p<0.01, p<0.01). In patients >75 years old receiving probucol, the relative risk (95% confidence interval) was 0.18 (0.02 to 1.44) for all-cause mortality and 0.14 (0.02 to 0.107) for major coronary events. In patients >75 years old receiving pravastatin, however, there were no significant differences of lelative risk compared with the control group. This FAST substudy showed that probucol was effective at reducing the incidence of cardiovasculat" disease in elderly hypercholesterolemic patients as well as younger patients.
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a fi'action of the patients needing U'eatment in primary care seem to be recognized and receive adequate treatrnent. The epidemiological study DETECT (Diabetes Cardiovascular Risk Evaluation: Tat'gets and Essential Data for Commitment of Treatment) was launched to identify the reasons, the extent and the short-terrn consequences of unmet needs in patients with high cat'diovasculat" risk. DETECT is a lat'ge multistage cross-sectional and prospective 12-month study in over 3000 primat'y cat'e offices. In stage 1, a mailed questionnah'e survey of physicians (N=3,572) and settings was pefforrned to assess physicians' awat'eness, attitudes, and practice patterns concerning CAD, arterial hypertension, diabetes mellitus, lipid disorders etc. In stage 2, a cross sectional study of consecutive patients (N=70,000 patients) in these primary care settings was completed using standardized questionnah'es for physicians and patients. In stage 3, a subset of 7,500 patients characterized by an extensive standardized laboratory program, are being followed up over 12 months to evaluate the change of selected laboratory measures and critical outcomes such as death, cat'diovasculat" events or hospitalisation. Aims of the study were to assess (a) the fi'equency, chat'acteristics and severity of the above diseases and selected comorbidites, (b) the proportion of patients with high-risk (c) rates of General Ih'actitioner's recognition, diagnoses and therapy, (d) quality of cale (e) indicators of undera'eatment, or over-treatment, respectively, and (e) to evaluate cma'ent laboratory measures. This presentation will report on the findings fi'om stage 1 and stage 3 with particular focus on the treatment modalities of lipid disorders in patients with coronary or vascular disease and/or type 2 diabetes mellitus.
THE GAP BETWEEN TREATMENT GUIDELINES AND ROUTINE CARE TREATMENT PATTERS IN THE MANAGEMENT OF H I G H RISK PATIENTS: FINDINGS FROM THE DETECT STUDY
H. Scharnagl, F. Freisinger, A. Tfl'an, T. Stojakovic, D. Pittrow, H. Glaesmer, S. Boehler, G. Ruf, W. Maerz, H. Wittchen. Clinical Institute
of Medical and Chemical Laboratory Diagnostics, Graz, Austria; Institut f r Klinische Pharmakologie, Pfizer GmbH, Dresden, Germany Numerous approaches to the primary and secondary prevention of cardiovascular diseases have been introduced in clinical routine care. Yet only
INtroduction: Oxidation of LDL is the first step in the development of atherosclerosis. Patients with diabetes have an excess risk of atherosclerotic morbidity and mortality. The aim of the present study was to examine relationships between levels of in vivo oxidized LDL (ox-LDL) and cat'otid intima-media thickness (IMT) and features of insulin resistance in subjects with- and without type 2 diabetes. Subjects and Methods: The study group consisted of 63 subjects with a normal glucose metabolism and 57 patients with well-controlled type 2 diabetes (mean HbAlc: 6.5 -4- 1.1%). Cfl'culating ox-LDL was measured in plasma by a recently developed competitive ELISA (Mercodia, Sweden). The cat'otid IMT was measured by ultrasound. Results: Cat'otid IMT was significantly higher in subjects with diabetes compared to controls (0.89 -4- 0.17 vs 0.83 -4- 0.11 ram; P < 0.05). Cfl'culating ox-LDL was significantly associated with IMT (P = 0.01), and ox-LDL was significantly increased in the highest tertile of carotid IMT compat'ed to the lowest and the intermediate tertile (68.7 -4- 19.5, 66.2 -4- 17.4, and 78.5 -4- 21.9 U/L, respectively, P < 0.01). Additionally, ox-LDL was positively related with HbAlc, insulin, waist cfl'cumference, SBP (P < 0.05, for all), and also with CRP, LDL-cholesterol, and plasma triglyceride (P < 0.001, for all). Conclusions: Cat'otid IMT was increased in subjects with type 2 diabetes and was positively associated with ox-LDL. Ox-LDL COla'elated also positively with featmes of the insulin resistance syndrome. These findings indicate that the cfl'culating ox-LDL level may contribute to the atherogenic process in subjects with diabetes.
•0•
NON-STENOTIC ECHOLUCENT PLAQUES IN THE CAROTID ARTERIES IN CLINICALLY HEALTHY, 58-YEAR OLD MEN PREDICT CARDIOVASCULAR EVENTS
C. Schmidt, B. Fagerberg, J. Wikstrand, J. Hulthe. Wallenberg Laboratory
for Cardiovascular Research, G teborg, Sweden Objective: To examine the relationship between plaque echogenicity, assessed by B-mode ultrasound, in the cat'otid at'teries and the development of clinical cardio- and cerebrovascular disease in a group of clinically healthy initially 58-year" old men during 6.6 years of follow-up. Background: Plaque echogenicity as assessed by B-mode ultrasound has been found to reliably pledict the content of soft tissue and the amount of calcification in carotid plaques. Results fi'om previous studies indicate that
74th EAS Congress, 17-20 April 2004, Seville, Spain
Miscellaneous echolucency of stenotic atherosclerotic plaques may be associated with an increased risk for cerebrovasculat" events. Methods: High-resolution B-mode ultrasound was used to chax'acterize plaque echogenicity in fore" subgroups - dominantly echolucent, substantially echolucent, dominantly echogenic and uniformly echogenic. Results: In the group without plaques 4 of 134 (2.9%) suffered fi'om an end-point (cat'diovasculax" death, non-fatal MI, non-fatal stroke or non-fatal aorta aneurysm) dm'ing follow-up. In the group with echogenic plaques no one suffered fi'om an end-point dm'ing follow-up, and in the group with echolucent plaques 12 of 130 (9.2%) suffeled fi'om an end-point during follow-up (p<0.05, for Uend). Conclusion: The data indicate that there is a trend towax'ds higher risk for CVD among initially healthy middle-aged men with relatively small, nonstenotic, echolucent plaques in the cat'otid at'tery as compax'ed to subjects with echogenic plaques or without plaques.
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NANOGEN T M MICROELECTRONIC CHIP FOR LARGE SCALE GENOTYPING
A. Sethi, A. Tybj~erg-Hansen, R. Andersen, B. Nordestgaat'd. Copenhagen
University Hospital, Herlev University Hospital, Copenhagen, Denmark Background: Single nucleotide polymorphisms (SNPs) can be detected by electronic ch'cuitry on NanogenTM 99 test sites silicon microchips. We examined throughput, accm'acy and cost-effectiveness of this method compax'ed with Restriction Fragment Length Polymorphism (RFLP). Methods: SNPs in the beta-2 adlenergic receptor gene, Argl6Gly, Gln27Glu, and Tbx164Ile, were analyzed in 3675 individuals by RFLE Using the same Polymerase Chain Reaction product a microchip assay was designed to detect all tbxee polymorphisms at once. Each sample was addressed to one of the hundred pads on the microchip. Hah'pins were neutralized by stabilizer oligos that hybridized to the suspected regions. A mix of pah-wise reporter probes labeled with cytochrome-3 or cytocbxome-5 fluoropholes were added for each SNP and fluorescence signal detected. In between adding of reporters for each mutation the microchip was denatm'ed by adding 0.1 mol/L NaOH. Results: To increase throughput, we developed two diffelent protocols by which 3 different SNPs were analyzed simultaneously on a single test site. Of 10,911 determinations, 6 were misclassified by the microelectronic chip (0.05%) and 3 by RFLP (0.03%)(p= 0.32). Total running cost per SNP genotyped using new microelectronic chips, reused microelectronic chips, and RFLP was gl.10, g0.40 and g0.88, respectively. Conclusions: We demonstrate that SNP detection using microelectronic chips is compax'able to RFLP with respect to tbxoughput, accm"
Siemianowicz
151
of patients took lipid-lowering drugs. Number of patients with tat'get LDL cholesterol consisted 44% fi'om all who finished obervation.
~
PARAOXONASE 2 EXPRESSION IS UP-REGULATED BY NADPH OXIDASE DURING MONOCYTE DIFFERENTIATION INTO MACROPHAGES
M. Shiner, B. Fuhrman, M. Avfl'am. The Lipid Research Laboratory,
Technion Faculty of Medicine, The Rappaport Family Institute for Research in the Medical Sciences and Rambam Medical Center, Haifa, Israel Pax'aoxonase 2 (PON2) is expressed in cells fi'om vax'ious tissues, including macrophages, and was lecently shown to possess antioxidant properties. In the present study we investigated the pattern and mechanisms responsible for PON2 expression dm'ing monocyte/macrophage- differentiation in vivo. PON2 mRNA expression and lactonase activity, increased, up-to 2.5 fold and up-to 4 fold, respectively during monocyte/macrophage differentiation in vivo, and were pax'alleled by an increase in cellulat" oxidative status. Supplementation of the antioxidant vitamin E to Balb/C mice inhibited the increase in cellulat" oxidative stress dm'ing macrophage matm'ation in-vivo by 50%, and in pax'allel, down-regulated PON2 expression and activity by 20%. The mechanism regulating PON2 expression could be related, at least in pax't, to NADPH-oxidase activation, since PON2 mRNA expression and lactonase activity in MPM isolated fi'om p47 ph°×-/- mice (inactive NADPHoxidase) were 9.0 fold and 5.5 fold lower than in MPM isolated fi'om Balb/C mice. Similax'ly, in vitro NADPH-oxidase activation was observed in PMA-treated THP-1 cell-line together with up-regulation of cellulm" PON2. These results of increased PON2 expression could be related to the activator protein 1 (AP-1) as inhibition of the AP-1 activator JNK resulted in a 20% decreased cellulat" PON2 expression. Fm'thermore, the expression of AP-1 repressor Jun dimerization protein (JDP2) decreased in the nucleus. We thus conclude that PON2 expression is increased during macrophage matm'ation. This phenomenon is related to NADPH oxidase enhancement of cellulat" oxidative stress that increases the transcription activity of AP-1, leading to elevated PON2 explession. Enhancement of macrophage PON2 expression may represent a compensatory mechanism against the development of atherosclerosis.
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ANTIOXIDANT STATUS AND ENDOTHELIUM DYSFUNCTION MARKERS IN PATIENTS WITH DIABETIC RETINOPATHY
K. Siemianowicz, J. Gminski, A. Telega, T. Francuz, M. Syzdol, A. Wojcik, B. Posielezna, R. Grabowska-Bochenek. Dept. of Biochemistry, Silesian
Medical Academy, Diabetic Outpatient Department, Katowice, Poland ~
EFFICACY OF MULTIFACTORIAL RISK FACTORS PREVENTIVE EFFORTS IN ACHIEVING TARGET LDL CHOLESTEROL LEVEL (< 115 mg/dl) IN SURVIVORS AFTER MYOCARDIAL INFARCTION
S. Shalaev, N. Dmitrieva, L. Kremneva, Z. Safiullin. Tyumen Cardiological
Center, Russia The aim of the work was to estimate the efficacy of achieving "tat'get" LDL cholesterol (< 115 mg/dl) during multifactorial risk factors preventive efforts. Material and methods: This investigation was opened compax'eable randomized prospective study of 2 groups of patients after myocat'dial infax'ction - the group of "active" management (n=88) and "observation" (n=48). All prophylactic measm'es had explanable and recommendational chax'acter. Achievement of tax'get LDL cholesterol, modified risk factors (smoking, at'terial hypertension, increased saturated fats and cholesterol consumption, orerweight and obesity), plasma lipid content were estimated tbxough 2-3, 5-6 and 12 monthes of observation. Multifactorial risk factors preventive efforts in the whole had positive influence on the modified risk factors. To the end of the observation the middle quantity of nonlipid risk factors for the 1 patient decreased fi'om 3.8-4-0.99 to 2.8-4-1.29 (p< 0.05). At the same time the achievement of the tax'get pat'ameters in many of the cases was unreal. About one half of the patients could not significantly decrease the consumption of saturated fat and cholesterol with the food. About 25% of the patients continued smoking. In 55% overweight and in 64% abdominal obesity was max'ked. In the group of "observation" to the end of the yeat" only 5 patients had LDL cholesterol < 115 mg/dl. Only 2 patients (4.2%) began lipidlowering therapy.In the group of active intervention the middle reduction of LDL cholesterol consisted 23%, 38%
Diabetes mellitus is considered as an equivalent of coronm'y heax't disease (CHD). Hyperglycaemia is one of factors causing endothelial damage. Adhesion molecules such as sVCAM-1, sICAM-1 and vWF factor as well as E-selectin are consideled as mm'kers of endothelial damage. Endothelial dysfunction in diabetes leads to an accelerated progression of atherosclerosis, which can clinically manifest as CHD, stroke and peripheral m'tery disease. Vasculat" complications of diabetes can affect also small vessels and manifests as diabetic retinopathy and nepbxopathy. Free radicals play an important role in pathogenesis of diabetes and atherosclerosis. ROS are annihilated by intracellulat" enzymatic system composed mainly of glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT). Beta-cax'otene, tocopherols and ascorbic acid ate major serum antioxidants. All serum antioxidants ate usually measured together as total serum antioxidant status (TAS). An imbalance between oxidative stress and antioxidant protective mechanisms may enhance a deleterious influence of ROS on arterial wall. The aim of ore" study was to measme the activity of antioxidant enzymes and TAS in patients with type 2 diabetes mellitus and to evaluate a COla'elation between them and sVCAM-1, sICAM-1, vWF factor and E-selectin. 20 patients with type 2 diabetes mellitus and retinopathy were introduced into the study. Patients with atherosclerotic lesions in internal cax'oid ax'tery detected in the ultrasound examination were excluded fi'om the study. In obtained blood samples erythrocyte activity of SOD, GPx and CAT were measm'ed according to manufactm'er recommendations. TAS was measm'ed in fi'ozen serum samples. Serum levels of sVCAM-1, sICAM-1, von Willebrand factor and E-selectin were measm'ed in duplicates according to manufactm'er recommendations. There were statistically significant negative COlVelations between TAS and sVCAM (r = -0,46); TAS and s-ICAM (r = -0,54) and between SOD and E-selectin (r = -0,52).
74th EAS Congress, 17-20 April 2004, Seville, Spain
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Miscellaneous
Sasiela EFFECT OF ATORVASTATIN ON LIPOPROTEIN SUBFRACTION PROFILES IN PATIENTS W I T H DYSLIPIDEMIA
W. Sasiela, H. Silbershatz, J. Ma, E. Villagio. Pfizer Inc, New York, USA
Background: A preponderance of small dense LDL is strongly associated with elevated CHD risk. Evidence fi'om small-scale, single-dose studies suggests that additional to reducing overall LDL-C, some statins can qualitatively change LDL subfi'action profile. Methods: NASDAC was a multicenter, double-blind, randomized trial comparing atorvastatin efficacy and safety at starting doses of 10, 20, 40 and 80mg. In a reU'ospective subgroup analysis, atorvastatin effects on various lipoprotein subfi'action profiles were determined using NMR technology. The subgroup population comprised 190 patients who were candidates for lipid-lowering therapy according to NCEP III criteria, had baseline TG >2.3mmol/L and who wele randomized to atorvastatin 10mg (n=40), 20mg (n=55), 40mg (n=47), or 80mg (n=48). Subfi'action analyses were conducted using NMR LipoIh'ofile ®. Results: In the subgroup population,atorvastatin significantly reduced LDL-C levels (measured by standat'd methods) at all doses (P<0.001; 10mg: -34%; 20mg: -40%; 40mg: -49%; 80mg: -52%). Reductions in LDL-C, TC and TG as measured by NMR were similar to those obtained by conventional methods. However, HDL increases were markedly higher when determined by NMR. At all doses, atorvastatin significantly leduced the proportion of LDL pat'ticles that ale small, dense LDL (-15% to -19%, P<0.005), reduced absolute number of LDL pat'ticles (P<0.0001), incleased LDL pat'ticle size (P<0.0001) and improved HDL-subfi'action profile. Conclusion: Atorvastatin beneficially altered the atherogenic lipid profile in dyslipidemia patients. As expected, atorvastatin significantly reduced LDL-C and LDL pat'ticle numbers. Furthermore, atorvastatin significantly decreased LDL pat'ticle density, resulting in a shift fi'om small, dense LDL to more buoyant and less atherogenic particles.
•]
OXIDIZED LDL IS ASSOCIATED WITH CAROTID IMT M•.649] AND FEATURES OF THE INSULIN RESISTANCE SYNDROME P.G. Scheffer, R.M.A. Henry, E.J.M. Wever, G. Bos, R.J. Heine, J.M. Dekker, C.D.A. Stehouwer, T. Teerlink. Institute for Cardiovascular
Research, VU University Medical Center, Amsterdam, The Netherlands EFFECTS OF PROBUCOL AND PRAVASTATIN IN HYPERCHOLESTEROLEMIC PATIENTS AGED 75 YEARS OR OLDER
Y. Sawayama, N. Maeda, M. Tatsukawa, C. Shimizu, J. Hayashi. Dept. of
General Medicine, Kyushu University Hospital, Fukuoka, Japan The plesent study involved post hoc assessment of the efficacy of lipidlowering therapy in two subsets of the Fukuoka Atherosclerosis Trial (FAST) patients (>75 years old or <75 yeat's old).The FAST cohort of 246 asymptomatic hypercholesterolemic patients (mean age: 66 yeat's) included 76 patients aged >75 years. Patients were randomized to receive probucol at 500 mg/day (n=82) or pravastatin at 10 mg/day (n=83), or were assigned to a control group treated by diet alone (n=81). The change of intima-media thickness (IMT) in the common carotid at'tery was the primary endpoint and the incidence of major cardiovascular events was the secondary endpoint.In patients >75 yeat's old, 2 yeat's of probucol or pravastatin therapy caused a significant decrease of IMT (by 5.8% and 5.5%, respectively; both p<0.05). These lipid-lowering drugs also significantly reduced the IMT in patients <75 yeat's old (by 18.7% and 13.3%, respectively; both p<0.01). In contrast, control patients >75 yeat's old showed a significant increase of IMT compaled with patients <75 yeat's old (by 34.3% and 12.1%, respectively; p<0.01, p<0.01). In patients >75 years old receiving probucol, the relative risk (95% confidence interval) was 0.18 (0.02 to 1.44) for all-cause mortality and 0.14 (0.02 to 0.107) for major coronary events. In patients >75 years old receiving pravastatin, however, there were no significant differences of lelative risk compared with the control group. This FAST substudy showed that probucol was effective at reducing the incidence of cardiovasculat" disease in elderly hypercholesterolemic patients as well as younger patients.
•]
a fi'action of the patients needing U'eatment in primary care seem to be recognized and receive adequate treatrnent. The epidemiological study DETECT (Diabetes Cardiovascular Risk Evaluation: Tat'gets and Essential Data for Commitment of Treatment) was launched to identify the reasons, the extent and the short-terrn consequences of unmet needs in patients with high cat'diovasculat" risk. DETECT is a lat'ge multistage cross-sectional and prospective 12-month study in over 3000 primat'y cat'e offices. In stage 1, a mailed questionnah'e survey of physicians (N=3,572) and settings was pefforrned to assess physicians' awat'eness, attitudes, and practice patterns concerning CAD, arterial hypertension, diabetes mellitus, lipid disorders etc. In stage 2, a cross sectional study of consecutive patients (N=70,000 patients) in these primary care settings was completed using standardized questionnah'es for physicians and patients. In stage 3, a subset of 7,500 patients characterized by an extensive standardized laboratory program, are being followed up over 12 months to evaluate the change of selected laboratory measures and critical outcomes such as death, cat'diovasculat" events or hospitalisation. Aims of the study were to assess (a) the fi'equency, chat'acteristics and severity of the above diseases and selected comorbidites, (b) the proportion of patients with high-risk (c) rates of General Ih'actitioner's recognition, diagnoses and therapy, (d) quality of cale (e) indicators of undera'eatment, or over-treatment, respectively, and (e) to evaluate cma'ent laboratory measures. This presentation will report on the findings fi'om stage 1 and stage 3 with particular focus on the treatment modalities of lipid disorders in patients with coronary or vascular disease and/or type 2 diabetes mellitus.
THE GAP BETWEEN TREATMENT GUIDELINES AND ROUTINE CARE TREATMENT PATTERS IN THE MANAGEMENT OF H I G H RISK PATIENTS: FINDINGS FROM THE DETECT STUDY
H. Scharnagl, F. Freisinger, A. Tfl'an, T. Stojakovic, D. Pittrow, H. Glaesmer, S. Boehler, G. Ruf, W. Maerz, H. Wittchen. Clinical Institute
of Medical and Chemical Laboratory Diagnostics, Graz, Austria; Institut f r Klinische Pharmakologie, Pfizer GmbH, Dresden, Germany Numerous approaches to the primary and secondary prevention of cardiovascular diseases have been introduced in clinical routine care. Yet only
INtroduction: Oxidation of LDL is the first step in the development of atherosclerosis. Patients with diabetes have an excess risk of atherosclerotic morbidity and mortality. The aim of the present study was to examine relationships between levels of in vivo oxidized LDL (ox-LDL) and cat'otid intima-media thickness (IMT) and features of insulin resistance in subjects with- and without type 2 diabetes. Subjects and Methods: The study group consisted of 63 subjects with a normal glucose metabolism and 57 patients with well-controlled type 2 diabetes (mean HbAlc: 6.5 -4- 1.1%). Cfl'culating ox-LDL was measured in plasma by a recently developed competitive ELISA (Mercodia, Sweden). The cat'otid IMT was measured by ultrasound. Results: Cat'otid IMT was significantly higher in subjects with diabetes compared to controls (0.89 -4- 0.17 vs 0.83 -4- 0.11 ram; P < 0.05). Cfl'culating ox-LDL was significantly associated with IMT (P = 0.01), and ox-LDL was significantly increased in the highest tertile of carotid IMT compat'ed to the lowest and the intermediate tertile (68.7 -4- 19.5, 66.2 -4- 17.4, and 78.5 -4- 21.9 U/L, respectively, P < 0.01). Additionally, ox-LDL was positively related with HbAlc, insulin, waist cfl'cumference, SBP (P < 0.05, for all), and also with CRP, LDL-cholesterol, and plasma triglyceride (P < 0.001, for all). Conclusions: Cat'otid IMT was increased in subjects with type 2 diabetes and was positively associated with ox-LDL. Ox-LDL COla'elated also positively with featmes of the insulin resistance syndrome. These findings indicate that the cfl'culating ox-LDL level may contribute to the atherogenic process in subjects with diabetes.
•0•
NON-STENOTIC ECHOLUCENT PLAQUES IN THE CAROTID ARTERIES IN CLINICALLY HEALTHY, 58-YEAR OLD MEN PREDICT CARDIOVASCULAR EVENTS
C. Schmidt, B. Fagerberg, J. Wikstrand, J. Hulthe. Wallenberg Laboratory
for Cardiovascular Research, G teborg, Sweden Objective: To examine the relationship between plaque echogenicity, assessed by B-mode ultrasound, in the cat'otid at'teries and the development of clinical cardio- and cerebrovascular disease in a group of clinically healthy initially 58-year" old men during 6.6 years of follow-up. Background: Plaque echogenicity as assessed by B-mode ultrasound has been found to reliably pledict the content of soft tissue and the amount of calcification in carotid plaques. Results fi'om previous studies indicate that
74th EAS Congress, 17-20 April 2004, Seville, Spain
Miscellaneous echolucency of stenotic atherosclerotic plaques may be associated with an increased risk for cerebrovasculat" events. Methods: High-resolution B-mode ultrasound was used to chax'acterize plaque echogenicity in fore" subgroups - dominantly echolucent, substantially echolucent, dominantly echogenic and uniformly echogenic. Results: In the group without plaques 4 of 134 (2.9%) suffered fi'om an end-point (cat'diovasculax" death, non-fatal MI, non-fatal stroke or non-fatal aorta aneurysm) dm'ing follow-up. In the group with echogenic plaques no one suffered fi'om an end-point dm'ing follow-up, and in the group with echolucent plaques 12 of 130 (9.2%) suffeled fi'om an end-point during follow-up (p<0.05, for Uend). Conclusion: The data indicate that there is a trend towax'ds higher risk for CVD among initially healthy middle-aged men with relatively small, nonstenotic, echolucent plaques in the cat'otid at'tery as compax'ed to subjects with echogenic plaques or without plaques.
•]
NANOGEN T M MICROELECTRONIC CHIP FOR LARGE SCALE GENOTYPING
A. Sethi, A. Tybj~erg-Hansen, R. Andersen, B. Nordestgaat'd. Copenhagen
University Hospital, Herlev University Hospital, Copenhagen, Denmark Background: Single nucleotide polymorphisms (SNPs) can be detected by electronic ch'cuitry on NanogenTM 99 test sites silicon microchips. We examined throughput, accm'acy and cost-effectiveness of this method compax'ed with Restriction Fragment Length Polymorphism (RFLP). Methods: SNPs in the beta-2 adlenergic receptor gene, Argl6Gly, Gln27Glu, and Tbx164Ile, were analyzed in 3675 individuals by RFLE Using the same Polymerase Chain Reaction product a microchip assay was designed to detect all tbxee polymorphisms at once. Each sample was addressed to one of the hundred pads on the microchip. Hah'pins were neutralized by stabilizer oligos that hybridized to the suspected regions. A mix of pah-wise reporter probes labeled with cytochrome-3 or cytocbxome-5 fluoropholes were added for each SNP and fluorescence signal detected. In between adding of reporters for each mutation the microchip was denatm'ed by adding 0.1 mol/L NaOH. Results: To increase throughput, we developed two diffelent protocols by which 3 different SNPs were analyzed simultaneously on a single test site. Of 10,911 determinations, 6 were misclassified by the microelectronic chip (0.05%) and 3 by RFLP (0.03%)(p= 0.32). Total running cost per SNP genotyped using new microelectronic chips, reused microelectronic chips, and RFLP was gl.10, g0.40 and g0.88, respectively. Conclusions: We demonstrate that SNP detection using microelectronic chips is compax'able to RFLP with respect to tbxoughput, accm"
Siemianowicz
151
of patients took lipid-lowering drugs. Number of patients with tat'get LDL cholesterol consisted 44% fi'om all who finished obervation.
~
PARAOXONASE 2 EXPRESSION IS UP-REGULATED BY NADPH OXIDASE DURING MONOCYTE DIFFERENTIATION INTO MACROPHAGES
M. Shiner, B. Fuhrman, M. Avfl'am. The Lipid Research Laboratory,
Technion Faculty of Medicine, The Rappaport Family Institute for Research in the Medical Sciences and Rambam Medical Center, Haifa, Israel Pax'aoxonase 2 (PON2) is expressed in cells fi'om vax'ious tissues, including macrophages, and was lecently shown to possess antioxidant properties. In the present study we investigated the pattern and mechanisms responsible for PON2 expression dm'ing monocyte/macrophage- differentiation in vivo. PON2 mRNA expression and lactonase activity, increased, up-to 2.5 fold and up-to 4 fold, respectively during monocyte/macrophage differentiation in vivo, and were pax'alleled by an increase in cellulat" oxidative status. Supplementation of the antioxidant vitamin E to Balb/C mice inhibited the increase in cellulat" oxidative stress dm'ing macrophage matm'ation in-vivo by 50%, and in pax'allel, down-regulated PON2 expression and activity by 20%. The mechanism regulating PON2 expression could be related, at least in pax't, to NADPH-oxidase activation, since PON2 mRNA expression and lactonase activity in MPM isolated fi'om p47 ph°×-/- mice (inactive NADPHoxidase) were 9.0 fold and 5.5 fold lower than in MPM isolated fi'om Balb/C mice. Similax'ly, in vitro NADPH-oxidase activation was observed in PMA-treated THP-1 cell-line together with up-regulation of cellulm" PON2. These results of increased PON2 expression could be related to the activator protein 1 (AP-1) as inhibition of the AP-1 activator JNK resulted in a 20% decreased cellulat" PON2 expression. Fm'thermore, the expression of AP-1 repressor Jun dimerization protein (JDP2) decreased in the nucleus. We thus conclude that PON2 expression is increased during macrophage matm'ation. This phenomenon is related to NADPH oxidase enhancement of cellulat" oxidative stress that increases the transcription activity of AP-1, leading to elevated PON2 explession. Enhancement of macrophage PON2 expression may represent a compensatory mechanism against the development of atherosclerosis.
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ANTIOXIDANT STATUS AND ENDOTHELIUM DYSFUNCTION MARKERS IN PATIENTS WITH DIABETIC RETINOPATHY
K. Siemianowicz, J. Gminski, A. Telega, T. Francuz, M. Syzdol, A. Wojcik, B. Posielezna, R. Grabowska-Bochenek. Dept. of Biochemistry, Silesian
Medical Academy, Diabetic Outpatient Department, Katowice, Poland ~
EFFICACY OF MULTIFACTORIAL RISK FACTORS PREVENTIVE EFFORTS IN ACHIEVING TARGET LDL CHOLESTEROL LEVEL (< 115 mg/dl) IN SURVIVORS AFTER MYOCARDIAL INFARCTION
S. Shalaev, N. Dmitrieva, L. Kremneva, Z. Safiullin. Tyumen Cardiological
Center, Russia The aim of the work was to estimate the efficacy of achieving "tat'get" LDL cholesterol (< 115 mg/dl) during multifactorial risk factors preventive efforts. Material and methods: This investigation was opened compax'eable randomized prospective study of 2 groups of patients after myocat'dial infax'ction - the group of "active" management (n=88) and "observation" (n=48). All prophylactic measm'es had explanable and recommendational chax'acter. Achievement of tax'get LDL cholesterol, modified risk factors (smoking, at'terial hypertension, increased saturated fats and cholesterol consumption, orerweight and obesity), plasma lipid content were estimated tbxough 2-3, 5-6 and 12 monthes of observation. Multifactorial risk factors preventive efforts in the whole had positive influence on the modified risk factors. To the end of the observation the middle quantity of nonlipid risk factors for the 1 patient decreased fi'om 3.8-4-0.99 to 2.8-4-1.29 (p< 0.05). At the same time the achievement of the tax'get pat'ameters in many of the cases was unreal. About one half of the patients could not significantly decrease the consumption of saturated fat and cholesterol with the food. About 25% of the patients continued smoking. In 55% overweight and in 64% abdominal obesity was max'ked. In the group of "observation" to the end of the yeat" only 5 patients had LDL cholesterol < 115 mg/dl. Only 2 patients (4.2%) began lipidlowering therapy.In the group of active intervention the middle reduction of LDL cholesterol consisted 23%, 38%
Diabetes mellitus is considered as an equivalent of coronm'y heax't disease (CHD). Hyperglycaemia is one of factors causing endothelial damage. Adhesion molecules such as sVCAM-1, sICAM-1 and vWF factor as well as E-selectin are consideled as mm'kers of endothelial damage. Endothelial dysfunction in diabetes leads to an accelerated progression of atherosclerosis, which can clinically manifest as CHD, stroke and peripheral m'tery disease. Vasculat" complications of diabetes can affect also small vessels and manifests as diabetic retinopathy and nepbxopathy. Free radicals play an important role in pathogenesis of diabetes and atherosclerosis. ROS are annihilated by intracellulat" enzymatic system composed mainly of glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT). Beta-cax'otene, tocopherols and ascorbic acid ate major serum antioxidants. All serum antioxidants ate usually measured together as total serum antioxidant status (TAS). An imbalance between oxidative stress and antioxidant protective mechanisms may enhance a deleterious influence of ROS on arterial wall. The aim of ore" study was to measme the activity of antioxidant enzymes and TAS in patients with type 2 diabetes mellitus and to evaluate a COla'elation between them and sVCAM-1, sICAM-1, vWF factor and E-selectin. 20 patients with type 2 diabetes mellitus and retinopathy were introduced into the study. Patients with atherosclerotic lesions in internal cax'oid ax'tery detected in the ultrasound examination were excluded fi'om the study. In obtained blood samples erythrocyte activity of SOD, GPx and CAT were measm'ed according to manufactm'er recommendations. TAS was measm'ed in fi'ozen serum samples. Serum levels of sVCAM-1, sICAM-1, von Willebrand factor and E-selectin were measm'ed in duplicates according to manufactm'er recommendations. There were statistically significant negative COlVelations between TAS and sVCAM (r = -0,46); TAS and s-ICAM (r = -0,54) and between SOD and E-selectin (r = -0,52).
74th EAS Congress, 17-20 April 2004, Seville, Spain