- Email: [email protected]
66 RESIDUAL PLATELET REACTIVITY ON DUAL ANTIPLATELET TREATMENT IS ASSOCIATED WITH ISCHEMIC EVENTS IN PATIENTS WITH ACUTE CORONARY SYNDROME: FROM A BIOLOGICAL TO A CLINICAL ENTITY
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Nutrition, Metabolism & Cardiovascular Diseases (2008) S35–S65
63 ANTIOXIDANT CAPACITY OF CIRCULATING ENDOTHELIAL PROGENITOR CELLS (EPCS) AND CIGARETTE SMOKING G. Mandraffino, A. Alibrandi, M. La Scala, C. Saitta, M.A. Sardo, S. Campo, F. Napoli, A. Saitta. Dipartimento di medicina interna e terapia medica, Policlinico universitario, Messina, Italy E-mail: giuseppemandraffi[email protected] Endothelial progenitor cells (EPCs) have been suggested to contribute to ongoing endothelial maintenance and repair. Decreased levels of circulating EPCs associated with increased formation of oxygen reactive species (ROS) have been reported in smoker subjects, however, the effects of cigarette smoking (CS) on EPCs remain to be clarified. In the present study we investigated the effects of CS on the levels of ROS and expression of manganese superoxide dismutase (MnSOD), catalase (CAT) and glutathione peroxidase type 1 (GPX-1)-mRNA in EPCs isolated from peripheral blood of 36 healthy cigarette smokers and 26 nonsmokers controls. Furthermore, we valuated the relationship between circulating EPCs and the plasma levels of CRP and endogenously NO-derived product nitrite/nitrate. In smokers the expression of MnSOD-mRNA was significantly higher (p < 0.001), whereas the expression of CAT and GPx-1-mRNA was lower (p < 0.001) than controls. The amount of ROS was more elevated in smokers (p < 0.001); by contrast the number of EPCs was reduced respect to controls (p < 0.001). A strong correlation between circulating EPCs and levels of CRP (rs = 0.939, p < 0.001), fibrinogen (rs = 0.632, p < 0.001) and nitrite/nitrate (rs= 0.880, p < 0.001) was observed. EPCs were also correlated with the amount of ROS (rs = 0.832, p < 0.001) and with the expression of MnSOD (rs = 0.859, p < 0.001), CAT (rs =0.926, p < 0.001) and GPx-1 (rs =0.927, p < 0.001) -mRNA. Moreover, significant correlations were observed between the levels of CRP, fibrinogen, nitrite/nitrate, and the expression of MnSOD (rs= 0.892, p < 0.001; rs= 0.688, p < 0.001; rs = 0.821, p < 0.001, respectively), CAT (rs =-0.907, p < 0.001; rs = 0.656, p < 0.001; rs= 0.844, p < 0.005, respectively), and GPx1-mRNA (rs = 0.863, p < 0.001; rs = 0.585, p < 0.001; rs= 0.863, p < 0.001). The correlation panel showed significant values between ROS and MnSODmRNA (rs= 0.827, p < 0.001), and between ROS and both CAT (rs = 0.795, p < 0.001), and GPx-1 (rs= 0.745, p < 0.001). Additionally, dependence analysis indicated that CS influenced positively the levels of CRP (p < 0.001), fibrinogen (p < 0.001) and ROS (p < 0.001) and negatively the levels of HDL-C (p < 0.05) and nitrite/nitrate (p < 0.001), according to the intensity of smoke exposure. Multivariate regression model for EPC, showed that circulating EPCs number depends on GPx-1 expression (p < 0.001), CRP (p < 0.001) and HDL-C (p < 0.005) plasma concentrations. Taken together our findings provide evidence for a new effect of smoke exposure involving the expression of EPCs antioxidant enzymes. Furthermore, they indicate that in smokers the inflammatory state and molecules may play a role in modulating EPCs levels and antioxidant enzymes. 64 THE ATHEROGENIC LIPOPROTEIN PHENOTYPE AND LDL SIZE AND SUBCLASSES IN WOMEN WITH GESTATIONAL DIABETES M. Rizzo1 , K. Berneis2 , A.E. Altinova3 , I. Pepe1 , L. Manna1 , G. Di Fede1 , M. Arslan3 , G.A. Spinas2 , G.B. Rini1 . 1 Department of Clinical Medicine and Emerging Diseases, University of Palermo, Italy, 2 Clinics for Endocrinology, Diabetes & Clinical Nutrition, University Hospital Zurich, Switzerland, 3 Department of Endocrinology and Metabolism, Gazi University, Faculty of Medicine, Ankara, Turkey E-mail: [email protected] Objective: Women with gestational diabetes are more likely to develop type-2 diabetes and cardiovascular diseases after pregnancy; yet, the exact type of their lipid alterations is still not fully clear. We investigated in women with gestational diabetes low-density lipoproteins (LDL) size and all seven subclasses, as well as the “atherogenic-lipoprotein-phenotype” (ALP, e.g. concomitant presence of elevated triglycerides, reduced HDL-cholesterol and increased small dense LDL). Design and Methods: In 27 women with gestational diabetes and 23 healthy pregnant women matched for age, weeks of gestation and body mass index (as controls) we measured plasma lipids and LDL size and subclasses by gradient gel electrophoresis between 24th and 28th weeks of gestation. Results: Although no significant differences were found in all plasma lipids, compared to controls women with gestational diabetes showed lower LDL size (p = 0.0007) due to reduced LDL-I (p = 0.0074) and increased LDL-IVA (p = 0.0146) and -IVB (p < 0.0001) subclasses. Correlation analysis revealed that fasting glucose, HOMA and glycated haemoglobin were inversely correlated with LDL-I and positively with LDL-IVA and -IVB (all p < 0.05). No prevalence of ALP was found in both groups due to high HDL-cholesterol levels, while elevated small, dense LDL were more prevalent in women with gestational diabetes than controls (33% vs 4%, p = 0.0107). Conclusions: Beyond plasma lipids, increased levels of small, dense LDL seem to be common in women with gestational diabetes. Yet, whether these findings affect the atherogenic process and the clinical endpoints in this category of subjects remains to be determined by future prospective studies.
65 INFLUENCE OF VISCERAL FAT ON METABOLIC PROFILE AND EARLY VASCULAR DAMAGE IN DYSLIPIDEMIC PATIENTS M.R. Mannarino, R. Helou, D. Covelli, G. Pucci, G. Schillaci, M. Pirro. Internal Medicine, Angiology and Arteriosclerosis Diseases. University of Perugia, Italy E-mail: [email protected] Aim: Visceral fat accumulation may induce metabolic disorders such as atherogenic dyslipidemia, hyperglycemia and hypertension and is associated with accelerated atherosclerosis and increased cardiovascular risk. Assessment of general fat and abdominal fat by traditional anthropometric measures, like BMI and waist circumference, often gives an approximate and sometimes unreliable estimate of visceral fat deposition. Conversely, ultrasonography has been suggested as a novel and reliable non-invasive technique to quantify visceral fat. Whether ultrasound-estimated visceral fat has an influence on metabolic profile and early markers of atherosclerosis, like arterial stiffness and endothelial dysfunction, above that of BMI and waist circumference is still unknown. Methods: Visceral fat area (VFA) was measured in seventy-five dyslipidemic patients by ultrasonography as follows: [VFA] = 9.008 + 1.191×[distance between the internal surface of the abdominal muscle and the splenic vein (mm)] + 0.978×[distance between the internal surface of the abdominal muscle and the posterior wall of the aorta at the umbilicus (mm)] + 3.644×[thickness of the fat layer of the posterior right renal wall (mm)]. All patients underwent measurement of aortic pulse wave velocity (aPWV) as an index of arterial stiffness and brachial flow-mediated vasodilation (FMV) as a measure of endothelial function. Results: VFA was positively correlated with plasma triglycerides (r = 0.502, p 0.001) and glucose (r = 0.459, p = 0.002) and negatively with HDL cholesterol levels (r = 0.434, p = 0.003). VFA above the median value (170 cm2 ) was associated with higher triglycerides and lower HDL in both patients with BMI 27.0 and BMI > 27.0. Significant correlations were found between visceral fat area and aPWV (r = 0.38; p = 0.004), and FMV (r = 0.370; p = 0.01). Similarly, BMI and waist circumferences were significant positive correlates of aPWV (r = 0.345, p = 0.009; r = 0.399, p = 0.002, respectively). Multivariate analysis including alternatively BMI, waist circumference and VFA as independent variables, along with significant covariates of aPWV (e.g. age, systolic blood pressure), showed that only visceral fat area was able to predict an increased aPWV (beta = 0.23, p = 0.04). Similarly, multivariate analysis including FMV as dependent variable and age, systolic blood pressure, LDL cholesterol, smoking status and either waist circumference or VFA as independent variables showed that VFA (beta = 0.39, p = 0.03) but not waist circumference was a significant predictor of FMV variability. Forcing both VFA and waist circumference in the multivariate model, confirmed VFA as a significant FMV negative covariate (beta = 0.70, p = 0.03). Conclusions: Among dyslipidemic patients increased visceral fat area estimated by ultrasound is a major determinant of multiple unfavourable metabolic abnormalities. Moreover, expansion of visceral fat depots contributes to arterial stiffness and reduced flow mediated vasodilation, over and above the deleterious influence of traditional measures of general and abdominal adiposity like BMI and waist circumference. Hence, a selective measurement of visceral fat may help to identify those dyslipidemic patients with a more deteriorated metabolic profile and a more compromised arterial function. 66 RESIDUAL PLATELET REACTIVITY ON DUAL ANTIPLATELET TREATMENT IS ASSOCIATED WITH ISCHEMIC EVENTS IN PATIENTS WITH ACUTE CORONARY SYNDROME: FROM A BIOLOGICAL TO A CLINICAL ENTITY R. Marcucci, A. Cordisco, A.M. Gori, B. Giusti, R. Paniccia, E. Antonucci, N. Maggini, G.F. Gensini, R. Abbate. Dipartimento di Area Critica Medico Chirurgica, Universit` a di Firenze, Italy E-mail: rossella.marcucci@unifi.it A dual antiplatelet regimen of aspirin plus clopidogrel is the standard treatment of patients with acute coronary syndromes (ACS) undergoing pecutaneous coronary revascularization (PCI) with stent implantation. A growing body of evidence, obtained in chronic cardiovascular patients as well as in ACS, is demonstrating that the biological entity of the residual platelet reactivity (RPR) despite dual antiplatelet treatment is associated with an increased risk of aderse cardiovascular events. This is the largest prospective study planned to demonstrate the clinical impact of RPR by different agonists (arachidonic acid AA, ADP and collagen) on the occurrence of major adverse coronary events in the setting of ACS. We have enrolled 1112 ACS patients (961 M/151 F; age: 69 (39 94) yrs) undergoing PCI on dual antiplatelet therapy. RPR has been defined as maximal platelet aggregation by 1 mM arachidonic acid ,10 mM ADP >70% and 2 mg/mL collagen 56% on venous blood samples obtained within 24 hrs from PCI. At a median follow-up of 8 months (1 48) MACE, including cardiac death, myocardial infarction (MI) and target lesion revascularization (TLR) for symptomatic restenosis, were recorded in 202 patients (18.1%): 24 (2.1%) cardiac deaths, 55 (4.9%) MI and 147 (13.2%) TLR. At univariate analysis, RPR by ADP was significantly associated with cardiac death [OR 4.09 (1.7 9.5), p < 0.001] and MI [OR 1.98 (1.01 3.8), p < 0.05] whereas RPR by AA and collagen were significantly associated with MI [OR 2.6 (1.5 4.6), p < 0.001 and OR 2.09 adjusted for (1.05 4.1), p < 0.05, respectively]. At multivariate analysis
Abstracts, XXII National Congress of the Italian Society for the Study of Arteriosclerosis (SISA) age, sex, hypertension, diabetes, smoking habit, dyslipidemia, BMI, systolic left ventricular function and renal function RPR by ADP and collagen were independent predictors of MACE [OR 1.6 (1.03 2.7), p < 0.05 and OR 1.6 (1.01 2.6), p < 0.05, respectively]. These results, obtained in a large number of patients, demonstrate that RPR is a clinical entity associated with the future occurrence of ischemic cardiac events. In particular, we found that RPR by ADP is a predictor of both cardiac deaths and MI and RPR by AA and collagen are predictors of MI. These data pave the way to future studies addressed to evacuate the posale clinical benefits of a tailored antiplatelet therapy in the setting of ACS. 67 CHOLESTEROL METABOLISM AND OBESITY IN TYPE 2 DIABETES: ROLE OF PLASMA STEROLS I. Marini, P. Bertucci, S. Zagari, M.R. Bollea, A. Lala. Department of Internal Medicine, University “Tor Vergata”, Rome, Italy E-mail: [email protected] Obesity and type 2 diabetes are two clinical conditions caracterized by a reduction of cholesterol absorption. We have studied plasmatic levels of two indexes of cholesterol synthesis (lathosterol and desmosterol) and three indexes of cholesterol absorption (sitosterol, campesterol and colestanol) in 52 normal subjects (N) and in 52 type 2 diabetic subjects (D) with good metabolic control, treated with diet and/or oral hypoglycaemic agents and a BMI = 28.8±5.4 kg/m2 . Both groups were administred a diet with similar amounts of phytosterols. In N there was an inverse correlation between synthesis and absorption indexes, while in D such correlation was lacking. In N there was a direct and significant correlation between lathosterol and total cholesterol, LDL cholesterol and triglycerides and an inverse correlation with HDL cholesterol; a direct and significant correlation between the absorption indexes and HDL cholesterol and an inverse correlation with triglycerides. In D there was a direct and significant correlation between lathosterol, sitosterol, campesterol and total cholesterol, HDL cholesterol, LDL cholesterol (p = 0.01 at least) and between lathosterol and BMI (p = 0.05). We then divided the D patients in two groups: GROUP A: BMI < 30 kg/m2 ; GROUP B: BMI > 30 kg/m2 . There were no differences between the two groups in synthesis and absorption indexes. In the B group there was an inverse and significant correlation between lathosterol and colestanol (P = 0.006) and desmosterol and colestanol (p = 0.007) and a loss of the correlation between lathosterol, total cholesterol, LDL cholesterol and triglycerides what was observed in the A group. In conclusion our study demonstrates in D: (1) the loss of normal inverse relationship between synthesis and absorption only in non obese; (2) direct correlation between cholesterol synthesis, lipid profile and BMI; (3) loss of direct correlation between hepatic cholesterol synthesis and lipid profile in obese. 68 CEREBROTENDINOUS XANTHOMATOSIS IN A COMPOUND HETEROZYGOTE FOR TWO MUTATIONS ON CYP27 GENE: DIAGNOSIS AND TREATMENT S. Martini1 , S. Calandra2 , M. Del Puppo3 , I. Cortella1 , C. Priore Oliva2 , C. Gabelli4 , G. Realdi1 , E. Manzato1 . 1 Dip. Scienze Mediche-Chirurgiche, Clinica Medica I, Universit` a di Padova; 2 Dip. Scienze Biomediche, Patologia Generale, Universit` a Modena/Reggio Emilia; 3 Dip. Medicina Sperimentale, a di Universit` a Milano-Bicocca; 4 Centro Invecchiamento Cerebrale, Universit` Padova, Italy E-mail: [email protected] A 50 year old female attended our Lipid Clinic for hypercholesterolemia and tendon xanthomas. In the past she had surgical removal of the left achilles xanthoma and of xanthelasma. Following multiple attacks of tendonitis, she was referred to an orthopaedic consultant and then to our clinic; she showed impressive xanthomata at both achilles tendons, at the patellar tendon insertion, at the back of the right elbow, and bilateral xanthelasma. Lipid values were: TC 325, TG 141, HDL-C 100, LDL-C 197 mg/dl. Neuropsychological evaluation showed minimal cognitive impairment (MMSE 26,99). Brain MRI demonstrated hyperintensity of the dentate nuclei, similar but milder abnormalities of the white matter, increased subaracnoideal spaces with mild cerebellar atrophy; brain SPET confirmed the findings. Plasma cholestanol was 3893 mg/dl (control = 110), while plant sterols (b-sitosterol and campesterol) were only slightly increased. Molecular analysis of CYP27 gene showed an heterozygosity for two different mutations, one on exon 3 (645G>C), determining an aminoacid substitution (A183P), already described as a cause of Cerebrotendinous Xanthomatosis (CTX) in homozygosity or compound heterozygosity, the second on exon 9 (1559G>A) determining an aminoacid substitution (R480H) never described up to now. After one year of treatment with simvastatin 40 mg/day and chenoursodeoxycholic acid 250 mg t.i.d. LDL-C levels decreased by 57% while plasma cholestanol declined by 80% with decreased volume of xanthelasma and no attacks of tendonitis. In conclusion, we described a case of CTX depending on compound heterozygosity for two different mutations of CYP27 gene, one well known and the other of new description, determining a phenotype similar to homozygotes but milder and late-onset. The results of drug treatment look promising, but only longer follow-up might clarify if long-term drug treatment prevents the possible severe complications of the disease in this subject.
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69 SIMULTANEOUS DETERMINATION OF SPHYNGOMYELIN AND FREE CERAMIDE IN BIOLOGICAL SAMPLES BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY M. Min` a1 , D. Noto1 , G. Barraco1 , G. Ferraro1 , F. Monteleone1 , S. Petta2 , A. Craxi2 , M.R. Averna1 . 1 Dept. of Clinical Medicine and Emerging Diseases and 2 Gastroenterology Unit, University of Palermo, Italy E-mail: [email protected] The sphingomyelin (SM) pathway is a signalling system that is conserved from yeast to humans. Ceramide (CER), the central molecule in this pathway, serves as a second messenger for several cellular functions. In particular, CER has been linked to obesity, insulin resistance and lipotoxicity. Sphingolipid production in animal tissues is largely dependent upon their de novo biosynthesis and it depends largely on the availability of longchain saturated fats, in the initial, rate-limiting step in de novo ceramide synthesis. CER can be obtained also by SM in particular conditions (e.g. death receptor agonists, ionizing radiation, oxidative stress, etc.) acutely triggering ceramide accumulation by activating various forms of sphingomyelinase. CER accumulates in insulin resistant tissues and is able to modulate the Insulin receptor signalling cascade inhibiting phosphorylation and activation of Akt/PKB; CER probably impairs also the mitocondrial function. CER and SM are classes of molecules differing for the linked acyl chains. GC/MS is able to discriminate single CER and/or SM by their molecular weight and degree of insaturation of the acyl chain. More, a high temperature induced fragmentation in the injector is able to distinguish CER from SM on the basis of a single sylilation in SM and double sylilation in CER, due to the available site of derivatization in CER occupied by a phosphorylcholine head in SM. In this study we describe the optimization of the method in GC/MS using a single ion monitoring (SIM) MS mode. Using true standards and crude ceramide extracts we were able to measure the percent distribution of SM species in the plasma. We were able to measure CER and SM distributions in cell cultures (free growing HepG2, HEK293 cells) and liver biopsies. We can also measure the absolute levels of the main CER (CER16:0 and CER24:1) in liver biopsies. 70 OXIDATIVE STRESS AND AORTIC VALVE STENOSIS F. Minardi1 , V. Cavalca1,2 , L. Dainese1 , F. Veglia1 , A. Guarino1 , M. Giroli1 , L. Boccotti2 , E. Tremoli1,3 . 1 Centro Cardiologico Monzino IRCCS, Milan; 2 Institute of Cardiology, University of Milan, Milan; 3 Department of Pharmacological Sciences, University of Milan, Milan, Italy E-mail: [email protected] Aortic valve stenosis (AVS) is a degenerative and progressive condition which shares some characteristics with atherosclerosis. Oxidative stress is involved in many degenerative diseases and in the progression of the atherosclerotic process. The association between oxidative stress and aortic fibrosis was documented in animal studies; however, to date, few data are available in humans. In this study we measured markers of oxidative damage (Free and Total Malondyhaldeide: F-, T-MDA, Advanced Oxidized Protein Products: AOPP, oxidized glutathione: GSSG) and of antioxidant defences (alfa and gammaTocopherol: alfa-, gamma-TH, reduced glutathione: GSH, total antioxidant capacity: IAC) to demonstrate an oxidative stress status in patients with aortic valve stenosis. Methods: Forty-three AVS patients undergoing valvular replacement (age: 67.8±10.5 yr, 70% males) and 29 healthy controls (age: 55.2±4.9 yr, 41% males) were enrolled. In AVS group, determinations were performed at hospital admission. Data were evaluated by analysis of covariance, adjusting for age, gender and BMI. Results: Both forms of MDA were higher in AVS patients than in controls (FMDA mean±SEM: 0.26±0.03 micromol/L and 0.21±0.01, p = 0.02; T-MDA: 1.74±0.04 and 1.54±0.1, p = 0.03 respectively). Levels of plasmatic alfaTH were lower in AVS patients in comparison with controls (mean±SEM: 11.1±0.8 microg/mL and 17.0±1.0, p < 0.0001; 33.8±2.6 and 44.5±3.2, p = 0.01 respectively). No significant differences were found in the other analytes considered. Conclusion: In aortic valve stenosis we evidenced an oxidative damage due to lipid peroxidation but not to protein oxidation. The observed depletion of alfa-TH suggests a potential benefit of vitamin E in the treatment of aortic valve stenosis progression. 71 EARLY CHOLESTEROL LOWERING TREATMENT IN ACUTE CORONARY SYNDROMES. A COMPARATIVE STUDY ATORVASTATIN 40 VS EZETIMIBE/SIMVASTATIN 10/20 L. Mircoli, M. Negrini, C. Turri, G. Protasoni, B. Brusoni, R. Seregni. Dip di Cardiologia e Interventistica Cardiovascolare, Fatebenefratelli Hospital, Milan, Italy E-mail: [email protected] Purpose: statin therapy reduces mortality in acute coronary syndrome (ACS). Whether this clinical advantage is mainly correlated to statin-dependent cholesterol (CHOL)-lowering effect rather than a CHOL independent effect is not still demonstrated. We compared in a pivotal study two different Chollowering pharmacological strategies early administered in patients affected by ACS. Methods: thirty patients admitted in our emergency department (ED) with ACS were consecutively randomised (1:1, open) to Atorvastatin 40 mg (A) vs
Nutrition, Metabolism & Cardiovascular Diseases (2008) S35–S65
63 ANTIOXIDANT CAPACITY OF CIRCULATING ENDOTHELIAL PROGENITOR CELLS (EPCS) AND CIGARETTE SMOKING G. Mandraffino, A. Alibrandi, M. La Scala, C. Saitta, M.A. Sardo, S. Campo, F. Napoli, A. Saitta. Dipartimento di medicina interna e terapia medica, Policlinico universitario, Messina, Italy E-mail: giuseppemandraffi[email protected] Endothelial progenitor cells (EPCs) have been suggested to contribute to ongoing endothelial maintenance and repair. Decreased levels of circulating EPCs associated with increased formation of oxygen reactive species (ROS) have been reported in smoker subjects, however, the effects of cigarette smoking (CS) on EPCs remain to be clarified. In the present study we investigated the effects of CS on the levels of ROS and expression of manganese superoxide dismutase (MnSOD), catalase (CAT) and glutathione peroxidase type 1 (GPX-1)-mRNA in EPCs isolated from peripheral blood of 36 healthy cigarette smokers and 26 nonsmokers controls. Furthermore, we valuated the relationship between circulating EPCs and the plasma levels of CRP and endogenously NO-derived product nitrite/nitrate. In smokers the expression of MnSOD-mRNA was significantly higher (p < 0.001), whereas the expression of CAT and GPx-1-mRNA was lower (p < 0.001) than controls. The amount of ROS was more elevated in smokers (p < 0.001); by contrast the number of EPCs was reduced respect to controls (p < 0.001). A strong correlation between circulating EPCs and levels of CRP (rs = 0.939, p < 0.001), fibrinogen (rs = 0.632, p < 0.001) and nitrite/nitrate (rs= 0.880, p < 0.001) was observed. EPCs were also correlated with the amount of ROS (rs = 0.832, p < 0.001) and with the expression of MnSOD (rs = 0.859, p < 0.001), CAT (rs =0.926, p < 0.001) and GPx-1 (rs =0.927, p < 0.001) -mRNA. Moreover, significant correlations were observed between the levels of CRP, fibrinogen, nitrite/nitrate, and the expression of MnSOD (rs= 0.892, p < 0.001; rs= 0.688, p < 0.001; rs = 0.821, p < 0.001, respectively), CAT (rs =-0.907, p < 0.001; rs = 0.656, p < 0.001; rs= 0.844, p < 0.005, respectively), and GPx1-mRNA (rs = 0.863, p < 0.001; rs = 0.585, p < 0.001; rs= 0.863, p < 0.001). The correlation panel showed significant values between ROS and MnSODmRNA (rs= 0.827, p < 0.001), and between ROS and both CAT (rs = 0.795, p < 0.001), and GPx-1 (rs= 0.745, p < 0.001). Additionally, dependence analysis indicated that CS influenced positively the levels of CRP (p < 0.001), fibrinogen (p < 0.001) and ROS (p < 0.001) and negatively the levels of HDL-C (p < 0.05) and nitrite/nitrate (p < 0.001), according to the intensity of smoke exposure. Multivariate regression model for EPC, showed that circulating EPCs number depends on GPx-1 expression (p < 0.001), CRP (p < 0.001) and HDL-C (p < 0.005) plasma concentrations. Taken together our findings provide evidence for a new effect of smoke exposure involving the expression of EPCs antioxidant enzymes. Furthermore, they indicate that in smokers the inflammatory state and molecules may play a role in modulating EPCs levels and antioxidant enzymes. 64 THE ATHEROGENIC LIPOPROTEIN PHENOTYPE AND LDL SIZE AND SUBCLASSES IN WOMEN WITH GESTATIONAL DIABETES M. Rizzo1 , K. Berneis2 , A.E. Altinova3 , I. Pepe1 , L. Manna1 , G. Di Fede1 , M. Arslan3 , G.A. Spinas2 , G.B. Rini1 . 1 Department of Clinical Medicine and Emerging Diseases, University of Palermo, Italy, 2 Clinics for Endocrinology, Diabetes & Clinical Nutrition, University Hospital Zurich, Switzerland, 3 Department of Endocrinology and Metabolism, Gazi University, Faculty of Medicine, Ankara, Turkey E-mail: [email protected] Objective: Women with gestational diabetes are more likely to develop type-2 diabetes and cardiovascular diseases after pregnancy; yet, the exact type of their lipid alterations is still not fully clear. We investigated in women with gestational diabetes low-density lipoproteins (LDL) size and all seven subclasses, as well as the “atherogenic-lipoprotein-phenotype” (ALP, e.g. concomitant presence of elevated triglycerides, reduced HDL-cholesterol and increased small dense LDL). Design and Methods: In 27 women with gestational diabetes and 23 healthy pregnant women matched for age, weeks of gestation and body mass index (as controls) we measured plasma lipids and LDL size and subclasses by gradient gel electrophoresis between 24th and 28th weeks of gestation. Results: Although no significant differences were found in all plasma lipids, compared to controls women with gestational diabetes showed lower LDL size (p = 0.0007) due to reduced LDL-I (p = 0.0074) and increased LDL-IVA (p = 0.0146) and -IVB (p < 0.0001) subclasses. Correlation analysis revealed that fasting glucose, HOMA and glycated haemoglobin were inversely correlated with LDL-I and positively with LDL-IVA and -IVB (all p < 0.05). No prevalence of ALP was found in both groups due to high HDL-cholesterol levels, while elevated small, dense LDL were more prevalent in women with gestational diabetes than controls (33% vs 4%, p = 0.0107). Conclusions: Beyond plasma lipids, increased levels of small, dense LDL seem to be common in women with gestational diabetes. Yet, whether these findings affect the atherogenic process and the clinical endpoints in this category of subjects remains to be determined by future prospective studies.
65 INFLUENCE OF VISCERAL FAT ON METABOLIC PROFILE AND EARLY VASCULAR DAMAGE IN DYSLIPIDEMIC PATIENTS M.R. Mannarino, R. Helou, D. Covelli, G. Pucci, G. Schillaci, M. Pirro. Internal Medicine, Angiology and Arteriosclerosis Diseases. University of Perugia, Italy E-mail: [email protected] Aim: Visceral fat accumulation may induce metabolic disorders such as atherogenic dyslipidemia, hyperglycemia and hypertension and is associated with accelerated atherosclerosis and increased cardiovascular risk. Assessment of general fat and abdominal fat by traditional anthropometric measures, like BMI and waist circumference, often gives an approximate and sometimes unreliable estimate of visceral fat deposition. Conversely, ultrasonography has been suggested as a novel and reliable non-invasive technique to quantify visceral fat. Whether ultrasound-estimated visceral fat has an influence on metabolic profile and early markers of atherosclerosis, like arterial stiffness and endothelial dysfunction, above that of BMI and waist circumference is still unknown. Methods: Visceral fat area (VFA) was measured in seventy-five dyslipidemic patients by ultrasonography as follows: [VFA] = 9.008 + 1.191×[distance between the internal surface of the abdominal muscle and the splenic vein (mm)] + 0.978×[distance between the internal surface of the abdominal muscle and the posterior wall of the aorta at the umbilicus (mm)] + 3.644×[thickness of the fat layer of the posterior right renal wall (mm)]. All patients underwent measurement of aortic pulse wave velocity (aPWV) as an index of arterial stiffness and brachial flow-mediated vasodilation (FMV) as a measure of endothelial function. Results: VFA was positively correlated with plasma triglycerides (r = 0.502, p 0.001) and glucose (r = 0.459, p = 0.002) and negatively with HDL cholesterol levels (r = 0.434, p = 0.003). VFA above the median value (170 cm2 ) was associated with higher triglycerides and lower HDL in both patients with BMI 27.0 and BMI > 27.0. Significant correlations were found between visceral fat area and aPWV (r = 0.38; p = 0.004), and FMV (r = 0.370; p = 0.01). Similarly, BMI and waist circumferences were significant positive correlates of aPWV (r = 0.345, p = 0.009; r = 0.399, p = 0.002, respectively). Multivariate analysis including alternatively BMI, waist circumference and VFA as independent variables, along with significant covariates of aPWV (e.g. age, systolic blood pressure), showed that only visceral fat area was able to predict an increased aPWV (beta = 0.23, p = 0.04). Similarly, multivariate analysis including FMV as dependent variable and age, systolic blood pressure, LDL cholesterol, smoking status and either waist circumference or VFA as independent variables showed that VFA (beta = 0.39, p = 0.03) but not waist circumference was a significant predictor of FMV variability. Forcing both VFA and waist circumference in the multivariate model, confirmed VFA as a significant FMV negative covariate (beta = 0.70, p = 0.03). Conclusions: Among dyslipidemic patients increased visceral fat area estimated by ultrasound is a major determinant of multiple unfavourable metabolic abnormalities. Moreover, expansion of visceral fat depots contributes to arterial stiffness and reduced flow mediated vasodilation, over and above the deleterious influence of traditional measures of general and abdominal adiposity like BMI and waist circumference. Hence, a selective measurement of visceral fat may help to identify those dyslipidemic patients with a more deteriorated metabolic profile and a more compromised arterial function. 66 RESIDUAL PLATELET REACTIVITY ON DUAL ANTIPLATELET TREATMENT IS ASSOCIATED WITH ISCHEMIC EVENTS IN PATIENTS WITH ACUTE CORONARY SYNDROME: FROM A BIOLOGICAL TO A CLINICAL ENTITY R. Marcucci, A. Cordisco, A.M. Gori, B. Giusti, R. Paniccia, E. Antonucci, N. Maggini, G.F. Gensini, R. Abbate. Dipartimento di Area Critica Medico Chirurgica, Universit` a di Firenze, Italy E-mail: rossella.marcucci@unifi.it A dual antiplatelet regimen of aspirin plus clopidogrel is the standard treatment of patients with acute coronary syndromes (ACS) undergoing pecutaneous coronary revascularization (PCI) with stent implantation. A growing body of evidence, obtained in chronic cardiovascular patients as well as in ACS, is demonstrating that the biological entity of the residual platelet reactivity (RPR) despite dual antiplatelet treatment is associated with an increased risk of aderse cardiovascular events. This is the largest prospective study planned to demonstrate the clinical impact of RPR by different agonists (arachidonic acid AA, ADP and collagen) on the occurrence of major adverse coronary events in the setting of ACS. We have enrolled 1112 ACS patients (961 M/151 F; age: 69 (39 94) yrs) undergoing PCI on dual antiplatelet therapy. RPR has been defined as maximal platelet aggregation by 1 mM arachidonic acid ,10 mM ADP >70% and 2 mg/mL collagen 56% on venous blood samples obtained within 24 hrs from PCI. At a median follow-up of 8 months (1 48) MACE, including cardiac death, myocardial infarction (MI) and target lesion revascularization (TLR) for symptomatic restenosis, were recorded in 202 patients (18.1%): 24 (2.1%) cardiac deaths, 55 (4.9%) MI and 147 (13.2%) TLR. At univariate analysis, RPR by ADP was significantly associated with cardiac death [OR 4.09 (1.7 9.5), p < 0.001] and MI [OR 1.98 (1.01 3.8), p < 0.05] whereas RPR by AA and collagen were significantly associated with MI [OR 2.6 (1.5 4.6), p < 0.001 and OR 2.09 adjusted for (1.05 4.1), p < 0.05, respectively]. At multivariate analysis
Abstracts, XXII National Congress of the Italian Society for the Study of Arteriosclerosis (SISA) age, sex, hypertension, diabetes, smoking habit, dyslipidemia, BMI, systolic left ventricular function and renal function RPR by ADP and collagen were independent predictors of MACE [OR 1.6 (1.03 2.7), p < 0.05 and OR 1.6 (1.01 2.6), p < 0.05, respectively]. These results, obtained in a large number of patients, demonstrate that RPR is a clinical entity associated with the future occurrence of ischemic cardiac events. In particular, we found that RPR by ADP is a predictor of both cardiac deaths and MI and RPR by AA and collagen are predictors of MI. These data pave the way to future studies addressed to evacuate the posale clinical benefits of a tailored antiplatelet therapy in the setting of ACS. 67 CHOLESTEROL METABOLISM AND OBESITY IN TYPE 2 DIABETES: ROLE OF PLASMA STEROLS I. Marini, P. Bertucci, S. Zagari, M.R. Bollea, A. Lala. Department of Internal Medicine, University “Tor Vergata”, Rome, Italy E-mail: [email protected] Obesity and type 2 diabetes are two clinical conditions caracterized by a reduction of cholesterol absorption. We have studied plasmatic levels of two indexes of cholesterol synthesis (lathosterol and desmosterol) and three indexes of cholesterol absorption (sitosterol, campesterol and colestanol) in 52 normal subjects (N) and in 52 type 2 diabetic subjects (D) with good metabolic control, treated with diet and/or oral hypoglycaemic agents and a BMI = 28.8±5.4 kg/m2 . Both groups were administred a diet with similar amounts of phytosterols. In N there was an inverse correlation between synthesis and absorption indexes, while in D such correlation was lacking. In N there was a direct and significant correlation between lathosterol and total cholesterol, LDL cholesterol and triglycerides and an inverse correlation with HDL cholesterol; a direct and significant correlation between the absorption indexes and HDL cholesterol and an inverse correlation with triglycerides. In D there was a direct and significant correlation between lathosterol, sitosterol, campesterol and total cholesterol, HDL cholesterol, LDL cholesterol (p = 0.01 at least) and between lathosterol and BMI (p = 0.05). We then divided the D patients in two groups: GROUP A: BMI < 30 kg/m2 ; GROUP B: BMI > 30 kg/m2 . There were no differences between the two groups in synthesis and absorption indexes. In the B group there was an inverse and significant correlation between lathosterol and colestanol (P = 0.006) and desmosterol and colestanol (p = 0.007) and a loss of the correlation between lathosterol, total cholesterol, LDL cholesterol and triglycerides what was observed in the A group. In conclusion our study demonstrates in D: (1) the loss of normal inverse relationship between synthesis and absorption only in non obese; (2) direct correlation between cholesterol synthesis, lipid profile and BMI; (3) loss of direct correlation between hepatic cholesterol synthesis and lipid profile in obese. 68 CEREBROTENDINOUS XANTHOMATOSIS IN A COMPOUND HETEROZYGOTE FOR TWO MUTATIONS ON CYP27 GENE: DIAGNOSIS AND TREATMENT S. Martini1 , S. Calandra2 , M. Del Puppo3 , I. Cortella1 , C. Priore Oliva2 , C. Gabelli4 , G. Realdi1 , E. Manzato1 . 1 Dip. Scienze Mediche-Chirurgiche, Clinica Medica I, Universit` a di Padova; 2 Dip. Scienze Biomediche, Patologia Generale, Universit` a Modena/Reggio Emilia; 3 Dip. Medicina Sperimentale, a di Universit` a Milano-Bicocca; 4 Centro Invecchiamento Cerebrale, Universit` Padova, Italy E-mail: [email protected] A 50 year old female attended our Lipid Clinic for hypercholesterolemia and tendon xanthomas. In the past she had surgical removal of the left achilles xanthoma and of xanthelasma. Following multiple attacks of tendonitis, she was referred to an orthopaedic consultant and then to our clinic; she showed impressive xanthomata at both achilles tendons, at the patellar tendon insertion, at the back of the right elbow, and bilateral xanthelasma. Lipid values were: TC 325, TG 141, HDL-C 100, LDL-C 197 mg/dl. Neuropsychological evaluation showed minimal cognitive impairment (MMSE 26,99). Brain MRI demonstrated hyperintensity of the dentate nuclei, similar but milder abnormalities of the white matter, increased subaracnoideal spaces with mild cerebellar atrophy; brain SPET confirmed the findings. Plasma cholestanol was 3893 mg/dl (control = 110), while plant sterols (b-sitosterol and campesterol) were only slightly increased. Molecular analysis of CYP27 gene showed an heterozygosity for two different mutations, one on exon 3 (645G>C), determining an aminoacid substitution (A183P), already described as a cause of Cerebrotendinous Xanthomatosis (CTX) in homozygosity or compound heterozygosity, the second on exon 9 (1559G>A) determining an aminoacid substitution (R480H) never described up to now. After one year of treatment with simvastatin 40 mg/day and chenoursodeoxycholic acid 250 mg t.i.d. LDL-C levels decreased by 57% while plasma cholestanol declined by 80% with decreased volume of xanthelasma and no attacks of tendonitis. In conclusion, we described a case of CTX depending on compound heterozygosity for two different mutations of CYP27 gene, one well known and the other of new description, determining a phenotype similar to homozygotes but milder and late-onset. The results of drug treatment look promising, but only longer follow-up might clarify if long-term drug treatment prevents the possible severe complications of the disease in this subject.
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69 SIMULTANEOUS DETERMINATION OF SPHYNGOMYELIN AND FREE CERAMIDE IN BIOLOGICAL SAMPLES BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY M. Min` a1 , D. Noto1 , G. Barraco1 , G. Ferraro1 , F. Monteleone1 , S. Petta2 , A. Craxi2 , M.R. Averna1 . 1 Dept. of Clinical Medicine and Emerging Diseases and 2 Gastroenterology Unit, University of Palermo, Italy E-mail: [email protected] The sphingomyelin (SM) pathway is a signalling system that is conserved from yeast to humans. Ceramide (CER), the central molecule in this pathway, serves as a second messenger for several cellular functions. In particular, CER has been linked to obesity, insulin resistance and lipotoxicity. Sphingolipid production in animal tissues is largely dependent upon their de novo biosynthesis and it depends largely on the availability of longchain saturated fats, in the initial, rate-limiting step in de novo ceramide synthesis. CER can be obtained also by SM in particular conditions (e.g. death receptor agonists, ionizing radiation, oxidative stress, etc.) acutely triggering ceramide accumulation by activating various forms of sphingomyelinase. CER accumulates in insulin resistant tissues and is able to modulate the Insulin receptor signalling cascade inhibiting phosphorylation and activation of Akt/PKB; CER probably impairs also the mitocondrial function. CER and SM are classes of molecules differing for the linked acyl chains. GC/MS is able to discriminate single CER and/or SM by their molecular weight and degree of insaturation of the acyl chain. More, a high temperature induced fragmentation in the injector is able to distinguish CER from SM on the basis of a single sylilation in SM and double sylilation in CER, due to the available site of derivatization in CER occupied by a phosphorylcholine head in SM. In this study we describe the optimization of the method in GC/MS using a single ion monitoring (SIM) MS mode. Using true standards and crude ceramide extracts we were able to measure the percent distribution of SM species in the plasma. We were able to measure CER and SM distributions in cell cultures (free growing HepG2, HEK293 cells) and liver biopsies. We can also measure the absolute levels of the main CER (CER16:0 and CER24:1) in liver biopsies. 70 OXIDATIVE STRESS AND AORTIC VALVE STENOSIS F. Minardi1 , V. Cavalca1,2 , L. Dainese1 , F. Veglia1 , A. Guarino1 , M. Giroli1 , L. Boccotti2 , E. Tremoli1,3 . 1 Centro Cardiologico Monzino IRCCS, Milan; 2 Institute of Cardiology, University of Milan, Milan; 3 Department of Pharmacological Sciences, University of Milan, Milan, Italy E-mail: [email protected] Aortic valve stenosis (AVS) is a degenerative and progressive condition which shares some characteristics with atherosclerosis. Oxidative stress is involved in many degenerative diseases and in the progression of the atherosclerotic process. The association between oxidative stress and aortic fibrosis was documented in animal studies; however, to date, few data are available in humans. In this study we measured markers of oxidative damage (Free and Total Malondyhaldeide: F-, T-MDA, Advanced Oxidized Protein Products: AOPP, oxidized glutathione: GSSG) and of antioxidant defences (alfa and gammaTocopherol: alfa-, gamma-TH, reduced glutathione: GSH, total antioxidant capacity: IAC) to demonstrate an oxidative stress status in patients with aortic valve stenosis. Methods: Forty-three AVS patients undergoing valvular replacement (age: 67.8±10.5 yr, 70% males) and 29 healthy controls (age: 55.2±4.9 yr, 41% males) were enrolled. In AVS group, determinations were performed at hospital admission. Data were evaluated by analysis of covariance, adjusting for age, gender and BMI. Results: Both forms of MDA were higher in AVS patients than in controls (FMDA mean±SEM: 0.26±0.03 micromol/L and 0.21±0.01, p = 0.02; T-MDA: 1.74±0.04 and 1.54±0.1, p = 0.03 respectively). Levels of plasmatic alfaTH were lower in AVS patients in comparison with controls (mean±SEM: 11.1±0.8 microg/mL and 17.0±1.0, p < 0.0001; 33.8±2.6 and 44.5±3.2, p = 0.01 respectively). No significant differences were found in the other analytes considered. Conclusion: In aortic valve stenosis we evidenced an oxidative damage due to lipid peroxidation but not to protein oxidation. The observed depletion of alfa-TH suggests a potential benefit of vitamin E in the treatment of aortic valve stenosis progression. 71 EARLY CHOLESTEROL LOWERING TREATMENT IN ACUTE CORONARY SYNDROMES. A COMPARATIVE STUDY ATORVASTATIN 40 VS EZETIMIBE/SIMVASTATIN 10/20 L. Mircoli, M. Negrini, C. Turri, G. Protasoni, B. Brusoni, R. Seregni. Dip di Cardiologia e Interventistica Cardiovascolare, Fatebenefratelli Hospital, Milan, Italy E-mail: [email protected] Purpose: statin therapy reduces mortality in acute coronary syndrome (ACS). Whether this clinical advantage is mainly correlated to statin-dependent cholesterol (CHOL)-lowering effect rather than a CHOL independent effect is not still demonstrated. We compared in a pivotal study two different Chollowering pharmacological strategies early administered in patients affected by ACS. Methods: thirty patients admitted in our emergency department (ED) with ACS were consecutively randomised (1:1, open) to Atorvastatin 40 mg (A) vs