- Email: [email protected]
20
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
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Ezetimibe 10 mg+Simvastatin 20 mg (E/S) administered within 12 hours after diagnosis of infarction S-T elevation (STEMI) or unstable angina (NSTEMI/UA) independently by CHOL levels. In ED were evaluated total CHOL and PCR considered as a non-lipidic biomarker. All patients were underwent to coronary angiography and angioplasty (according to procedure-timing guidelines). Total CHOL, LDL CHOL, HDL CHOL, Triglycerides (T) and PCR were repeated within 48 hours after admission, 4 weeks and 6 months later. Moreover, data of cardiac mortality and morbility were considered. Results: groups (A vs E/S) were homogeneous for age (years, meanSD, 61.1±14.0 vs 62.6±12.1), gender (M 10 vs 10; F 5 vs 5), risk factors (diabetes 4 vs 4; hypertension 8 vs 7), clinical diagnosis (STEMI 5 vs 6; NSTEMI/UA 9 vs 10), total COL (mg/dl, mean±±SD, 225.6±±43.3 vs 230.8±41.9 mg/dl, p = ns), LDL (127.2±20.5 vs 122.4±28.6, p = ns), HDL (42.3±13.5 vs 45.6±12.4, p = ns), T (148.5±63.1 vs 157.2±54.2, p = ns) and PCR (mg/l, 8.1±8.0 vs 7.9±7.4, p = ns). Four weeks later total CHOL (A vs E/S; 162.5±25.4, 31% vs 165.5±19.1, 31%, p = ns) and LDL (A vs E/S; 82.6±9.6, 35% vs 84.4±11.1, 36%, p = ns) were similarly reduced in both groups (p < 0.05 vs basal) and after 6 months as well (data not shown). No significant effects in HDL and T were observed in either group. PCR lowering effect was similar and non significant after 4 weeks in both groups (A 24% vs E/S 28%), whereas it was similar but significant (p < 0.05 vs basal) after 6 months (A 55% vs E/S 49%). No fatal events or morbidity differences (1 case each) during the follow-up. Conclusions: both early CHOL-lowering treatments effectively reduced Total and LDL CHOL in ACS. The similar effect on PCR seems to be correlated with absolute CHOL reduction (independently by pharmacological strategy) rather than a specific non-lipidic effect of statin. 72 SERUM LIPID PROFILE CHANGES ASSOCIATED WITH CANCER PROGRESSION Sandro Muntoni, L. Atzori, R. Mereu, G. Satta, M.D. Macis, M. Congia, A. Tedde, A. Desogus, Sergio Muntoni. University School of Cagliari and Centre For Metabolic Diseases, Cagliari, Italy E-mail: [email protected] Background and Aims: In solid and haematological tumours serum lipoprotein profile has been reported to be altered; decreased levels of total cholesterol and increased values of triglycerides have been found. Mechanisms and meaning of these changes are, however, not fully clarified. The aim of the present study was to determine relationships between cancer progression and serum lipoproteins. Methods and Results: We performed a case-control study and included cancer patients admitted to the 1st Division of Medical Oncology, Businco Hospital of Cagliari, Italy; 519 patients with any types of solid tumours, and 928 healthy controls. We considered total-Cholesterol (C), HDL-C, LDL-C, triglycerides and apolipoprotein A-1; other parameters examined were glycemia, insulinemia, Body Mass Index (BMI), homeostasis model assessment-estimated insulin resistance (HOMA-IR), C reactive protein (CRP) and tumor necrosis factor-a (TNF-a). In cancer group HDL-C and apolipoprotein A-1 were lower (p < 0.05) and triglycerides higher (p < 0.05) than in controls; HDL-C (mg/dl) females: 48 vs 64; males, 40 vs 52; Apo-A-1 (mg/dl) females: 125 vs 173; males, 120 vs 152; triglycerides (mg/dl) females: 133 vs 96; males, 152 vs 117. Glucose (mg/dl) was lower in cancer group (p < 0.05); females, 72.3 vs 80.0; males, 75.7 vs 78.4. Conclusion: Using a multivariate analysis, we were able to rule out cardiovascular and inflammatory diseases as causes of low HDL-C, and to demonstrate that these alterations can be exploited as specific consequences of the presence of a malignant tumour, with a diagnostic and prognostic significance. 73 DIFFERENTIAL EFFECTS OF LOW FAT AND LOW CARBOHYDRATE DIET IN THE MANAGEMENT OF METABOLIC SYNDROME IN OBESE NON DIABETIC PATIENTS F. Muzio1 , A. Boggio1 , M. Fiscella1 , L. Mondazzi1 , E. Passaro1 , C. Nalini1 , D. Sommariva1 , A. Branchi2 . 1 Department of Internal Medicine, G. Salvini Hospital, Garbagnate Milanese; 2 Department of Internal Medicine, University of Milan, Fondazione IRCCS Ospedale Maggiore, Policlinico, Milan, Italy E-mail: [email protected] There is full agreement that lifestyle changes primarily focused on weight reduction are the first-line approach to patients with metabolic syndrome. Nevertheless, the optimal diet for the metabolic syndrome still remains uncertain. Current recommendations support a diet containing 25 35% of calories as fats, 50 60% as carbohydrates and approximately 15% as proteins. However, decreasing dietary intake of carbohydrates has been repeatedly confirmed to lower fasting plasma triglycerides (TG) and have been reported to increase insulin sensitivity. This study was conducted to explore the hypothesis that hypocaloric diets of differing macronutrient composition would have differential effects on specific risk factors of the metabolic syndrome. The study was carried out in 115 obese non diabetic patients with metabolic syndrome as diagnosed according to the criteria of the International Diabetes Federation. The patients were randomly assigned to either a diet poor in fat (LF diet; carbohydrates 65%, protein 13%, total fat 22%, unsaturated fat 17%), or a diet lower in carbohydrates and higher in protein and monounsaturated
fat (LC diet; carbohydrates 48%, protein 19%, total fat 33%, unsaturated fat 24%). Both diets provided an approximately 500 kcalorie/day deficit based on each patient’s estimated daily energy expenditure. After 5 months, body weight, BMI, waist girth, systolic and diastolic blood pressure, total cholesterol, serum TG, blood glucose, insulin and HOMAIR (homeostasis model assessment) significantly decreased in both dietary groups, whereas HDL cholesterol did not change significantly from baseline. The mean percent changes from baseline of body weight, BMI, waist girth, diastolic blood pressure, total cholesterol, blood glucose, insulin and HOMAIR were similar in both dietary groups. The LC diet was associated with a greater decrease in systolic blood pressure and in serum TG level than the LF diet. Heart rate decreased only with the former and LDL-C only with the latter. Percent reduction of positive diagnostic criteria of metabolic syndrome in comparison to baseline were: central obesity 2% in LF and 10% in LC group (P=NS), hyperglycemia 29% in LF and 24% in LC group (P=NS), hypertension 9% in LF and 27% in LC group (P < 0.01), low HDL-C 2% in LF and 7% in LC group (P=NS), hypertriglyceridemia 23% in LF and 50% in LC group (P < 0.01). At the end of the study, 29% of patients in the LF and 49% in the LC group (P < 0.05) did not longer fulfilled the criteria of the metabolic syndrome. In conclusion, the extent of recovery from metabolic syndrome was significantly greater in LC than in LF dietary group. LC diet was associated with a greater decrease in the prevalence of hypertension and hypertriglyceridemia. LF diet significantly decreased LDL-C. This evidence suggests that tailoring of diet treatment to the specific metabolic and physical profile of each metabolic syndrome patient may be a fruitful approach. Carbohydrates would substitute for fat in those patients with high LDL-C, but a lower carbohydrate, higher protein and fat diet might be optimal for patients with high blood pressure and/or high TG. 74 INFLUENCE OF LDL AND ACTIVATION STATUS ON MACROPHAGE CHOLESTEROL ACCUMULATION M. Napolitano1 , A. Cantafora2 , E. Bravo1 . 1 Dept. Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanit` a, Rome; 2 Dept. of Clinical Medicine Gastroenterology, University “La Sapienza”, Policlinico Umberto I, Rome, Italy E-mail: [email protected] Modified LDL have a well known atherogenic role, but also levels of circulating native LDL may affect cholesterol accumulation in arteries. Macrophages are primary cell types with an active involvement in all phases of the atherogenesis, by both accumulating lipids and modulating the immuno-inflammatory response through their secretion of stimulating factors. Therefore, the connection between cholesterol deposition inside these cells and either levels of circulating LDL or macrophage activation status found many experimental evidences, as in this study where cholesterol accumulation in function of both LDL levels and macrophage activation status was evaluated. LDL were isolated by sequential ultracentrifugation from plasma of normo-lipidemic subjects by ultracentrifugation methods. Primary macrophages derived from human monocyte (HMDM) were activated by incubation with LPS (1 mg/ml) or phorbol esters (PMA, 0.5 mg/ml). After 24 h, resting (untreated) macrophage as well as activated cells, were incubated for further 24 hours with 0, 100, 1000 and 1500 mg/ml cholesterol carried by LDL. The incubations were performed in serum-free medium and control incubations were carried on resting cells in the absence of pro-activating factors (CT). Cholesterol accumulation, measured by a fluorescence method, increased in resting macrophage in function of LDL concentration in a dosedependent manner from 512+213 without LDL to 1163+ 433 mg/mg protein with 1500 mg/ml LDL (n = 4). The comparison between LPS- and PMA-treated macrophages and CT incubations, showed that cholesterol accumulated in the pro-inflamed cells was at least 2 times higher than CT cells, both in the absence and in the presence of LDL at any concentrations tested. However, differently from what observed in resting cells, in the pro-inflamed state the cholesterol accumulation was not correlated to the LDL concentrations. In fact, when expressed in term of cholesterol with respect to the cell incubated without LDL but treated with the same pro-stimulating factor, the entity of the increase in cholesterol content resulted to be more modestly increased in pro-inflamed cell than CT macrophages. On the whole, present results indicate that activation status and LDL concentration work as independent, not synergistic, factors in inducing the increase of cholesterol content in macrophages. 75 EFFECTS OF PCSK9 VARIANTS ON COMMON CAROTID ARTERY AND RELATION TO APoE ALLELES G.D. Norata1,2 , K. Garlaschelli2 , L. Grigore2 , S. Raselli2 , S. Tramontana2 , G. Buccianti3 , A.L. Catapano1,2 . 1 Department of Pharmacological Sciences, University of Milan; 2 Centre for the Study of Atherosclerosis, Italian Society for the Study of Atherosclerosis, Bassini Hospital, Cinisello Balsamo; 3 The Italian Society for the Prevention of Kidney Disease, ASPREMARE, Milan, Italy E-mail: [email protected] Context: PCSK9 plays a key role in plasma cholesterol metabolism by modulating the expression of LDL receptors. Objective and Patients: In this study we investigated the effects of two common polymorphism of the PCSK9 gene (E670G and I474V) on the intima
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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
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Nutrition, Metabolism & Cardiovascular Diseases (2008) S35–S65
Ezetimibe 10 mg+Simvastatin 20 mg (E/S) administered within 12 hours after diagnosis of infarction S-T elevation (STEMI) or unstable angina (NSTEMI/UA) independently by CHOL levels. In ED were evaluated total CHOL and PCR considered as a non-lipidic biomarker. All patients were underwent to coronary angiography and angioplasty (according to procedure-timing guidelines). Total CHOL, LDL CHOL, HDL CHOL, Triglycerides (T) and PCR were repeated within 48 hours after admission, 4 weeks and 6 months later. Moreover, data of cardiac mortality and morbility were considered. Results: groups (A vs E/S) were homogeneous for age (years, meanSD, 61.1±14.0 vs 62.6±12.1), gender (M 10 vs 10; F 5 vs 5), risk factors (diabetes 4 vs 4; hypertension 8 vs 7), clinical diagnosis (STEMI 5 vs 6; NSTEMI/UA 9 vs 10), total COL (mg/dl, mean±±SD, 225.6±±43.3 vs 230.8±41.9 mg/dl, p = ns), LDL (127.2±20.5 vs 122.4±28.6, p = ns), HDL (42.3±13.5 vs 45.6±12.4, p = ns), T (148.5±63.1 vs 157.2±54.2, p = ns) and PCR (mg/l, 8.1±8.0 vs 7.9±7.4, p = ns). Four weeks later total CHOL (A vs E/S; 162.5±25.4, 31% vs 165.5±19.1, 31%, p = ns) and LDL (A vs E/S; 82.6±9.6, 35% vs 84.4±11.1, 36%, p = ns) were similarly reduced in both groups (p < 0.05 vs basal) and after 6 months as well (data not shown). No significant effects in HDL and T were observed in either group. PCR lowering effect was similar and non significant after 4 weeks in both groups (A 24% vs E/S 28%), whereas it was similar but significant (p < 0.05 vs basal) after 6 months (A 55% vs E/S 49%). No fatal events or morbidity differences (1 case each) during the follow-up. Conclusions: both early CHOL-lowering treatments effectively reduced Total and LDL CHOL in ACS. The similar effect on PCR seems to be correlated with absolute CHOL reduction (independently by pharmacological strategy) rather than a specific non-lipidic effect of statin. 72 SERUM LIPID PROFILE CHANGES ASSOCIATED WITH CANCER PROGRESSION Sandro Muntoni, L. Atzori, R. Mereu, G. Satta, M.D. Macis, M. Congia, A. Tedde, A. Desogus, Sergio Muntoni. University School of Cagliari and Centre For Metabolic Diseases, Cagliari, Italy E-mail: [email protected] Background and Aims: In solid and haematological tumours serum lipoprotein profile has been reported to be altered; decreased levels of total cholesterol and increased values of triglycerides have been found. Mechanisms and meaning of these changes are, however, not fully clarified. The aim of the present study was to determine relationships between cancer progression and serum lipoproteins. Methods and Results: We performed a case-control study and included cancer patients admitted to the 1st Division of Medical Oncology, Businco Hospital of Cagliari, Italy; 519 patients with any types of solid tumours, and 928 healthy controls. We considered total-Cholesterol (C), HDL-C, LDL-C, triglycerides and apolipoprotein A-1; other parameters examined were glycemia, insulinemia, Body Mass Index (BMI), homeostasis model assessment-estimated insulin resistance (HOMA-IR), C reactive protein (CRP) and tumor necrosis factor-a (TNF-a). In cancer group HDL-C and apolipoprotein A-1 were lower (p < 0.05) and triglycerides higher (p < 0.05) than in controls; HDL-C (mg/dl) females: 48 vs 64; males, 40 vs 52; Apo-A-1 (mg/dl) females: 125 vs 173; males, 120 vs 152; triglycerides (mg/dl) females: 133 vs 96; males, 152 vs 117. Glucose (mg/dl) was lower in cancer group (p < 0.05); females, 72.3 vs 80.0; males, 75.7 vs 78.4. Conclusion: Using a multivariate analysis, we were able to rule out cardiovascular and inflammatory diseases as causes of low HDL-C, and to demonstrate that these alterations can be exploited as specific consequences of the presence of a malignant tumour, with a diagnostic and prognostic significance. 73 DIFFERENTIAL EFFECTS OF LOW FAT AND LOW CARBOHYDRATE DIET IN THE MANAGEMENT OF METABOLIC SYNDROME IN OBESE NON DIABETIC PATIENTS F. Muzio1 , A. Boggio1 , M. Fiscella1 , L. Mondazzi1 , E. Passaro1 , C. Nalini1 , D. Sommariva1 , A. Branchi2 . 1 Department of Internal Medicine, G. Salvini Hospital, Garbagnate Milanese; 2 Department of Internal Medicine, University of Milan, Fondazione IRCCS Ospedale Maggiore, Policlinico, Milan, Italy E-mail: [email protected] There is full agreement that lifestyle changes primarily focused on weight reduction are the first-line approach to patients with metabolic syndrome. Nevertheless, the optimal diet for the metabolic syndrome still remains uncertain. Current recommendations support a diet containing 25 35% of calories as fats, 50 60% as carbohydrates and approximately 15% as proteins. However, decreasing dietary intake of carbohydrates has been repeatedly confirmed to lower fasting plasma triglycerides (TG) and have been reported to increase insulin sensitivity. This study was conducted to explore the hypothesis that hypocaloric diets of differing macronutrient composition would have differential effects on specific risk factors of the metabolic syndrome. The study was carried out in 115 obese non diabetic patients with metabolic syndrome as diagnosed according to the criteria of the International Diabetes Federation. The patients were randomly assigned to either a diet poor in fat (LF diet; carbohydrates 65%, protein 13%, total fat 22%, unsaturated fat 17%), or a diet lower in carbohydrates and higher in protein and monounsaturated
fat (LC diet; carbohydrates 48%, protein 19%, total fat 33%, unsaturated fat 24%). Both diets provided an approximately 500 kcalorie/day deficit based on each patient’s estimated daily energy expenditure. After 5 months, body weight, BMI, waist girth, systolic and diastolic blood pressure, total cholesterol, serum TG, blood glucose, insulin and HOMAIR (homeostasis model assessment) significantly decreased in both dietary groups, whereas HDL cholesterol did not change significantly from baseline. The mean percent changes from baseline of body weight, BMI, waist girth, diastolic blood pressure, total cholesterol, blood glucose, insulin and HOMAIR were similar in both dietary groups. The LC diet was associated with a greater decrease in systolic blood pressure and in serum TG level than the LF diet. Heart rate decreased only with the former and LDL-C only with the latter. Percent reduction of positive diagnostic criteria of metabolic syndrome in comparison to baseline were: central obesity 2% in LF and 10% in LC group (P=NS), hyperglycemia 29% in LF and 24% in LC group (P=NS), hypertension 9% in LF and 27% in LC group (P < 0.01), low HDL-C 2% in LF and 7% in LC group (P=NS), hypertriglyceridemia 23% in LF and 50% in LC group (P < 0.01). At the end of the study, 29% of patients in the LF and 49% in the LC group (P < 0.05) did not longer fulfilled the criteria of the metabolic syndrome. In conclusion, the extent of recovery from metabolic syndrome was significantly greater in LC than in LF dietary group. LC diet was associated with a greater decrease in the prevalence of hypertension and hypertriglyceridemia. LF diet significantly decreased LDL-C. This evidence suggests that tailoring of diet treatment to the specific metabolic and physical profile of each metabolic syndrome patient may be a fruitful approach. Carbohydrates would substitute for fat in those patients with high LDL-C, but a lower carbohydrate, higher protein and fat diet might be optimal for patients with high blood pressure and/or high TG. 74 INFLUENCE OF LDL AND ACTIVATION STATUS ON MACROPHAGE CHOLESTEROL ACCUMULATION M. Napolitano1 , A. Cantafora2 , E. Bravo1 . 1 Dept. Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanit` a, Rome; 2 Dept. of Clinical Medicine Gastroenterology, University “La Sapienza”, Policlinico Umberto I, Rome, Italy E-mail: [email protected] Modified LDL have a well known atherogenic role, but also levels of circulating native LDL may affect cholesterol accumulation in arteries. Macrophages are primary cell types with an active involvement in all phases of the atherogenesis, by both accumulating lipids and modulating the immuno-inflammatory response through their secretion of stimulating factors. Therefore, the connection between cholesterol deposition inside these cells and either levels of circulating LDL or macrophage activation status found many experimental evidences, as in this study where cholesterol accumulation in function of both LDL levels and macrophage activation status was evaluated. LDL were isolated by sequential ultracentrifugation from plasma of normo-lipidemic subjects by ultracentrifugation methods. Primary macrophages derived from human monocyte (HMDM) were activated by incubation with LPS (1 mg/ml) or phorbol esters (PMA, 0.5 mg/ml). After 24 h, resting (untreated) macrophage as well as activated cells, were incubated for further 24 hours with 0, 100, 1000 and 1500 mg/ml cholesterol carried by LDL. The incubations were performed in serum-free medium and control incubations were carried on resting cells in the absence of pro-activating factors (CT). Cholesterol accumulation, measured by a fluorescence method, increased in resting macrophage in function of LDL concentration in a dosedependent manner from 512+213 without LDL to 1163+ 433 mg/mg protein with 1500 mg/ml LDL (n = 4). The comparison between LPS- and PMA-treated macrophages and CT incubations, showed that cholesterol accumulated in the pro-inflamed cells was at least 2 times higher than CT cells, both in the absence and in the presence of LDL at any concentrations tested. However, differently from what observed in resting cells, in the pro-inflamed state the cholesterol accumulation was not correlated to the LDL concentrations. In fact, when expressed in term of cholesterol with respect to the cell incubated without LDL but treated with the same pro-stimulating factor, the entity of the increase in cholesterol content resulted to be more modestly increased in pro-inflamed cell than CT macrophages. On the whole, present results indicate that activation status and LDL concentration work as independent, not synergistic, factors in inducing the increase of cholesterol content in macrophages. 75 EFFECTS OF PCSK9 VARIANTS ON COMMON CAROTID ARTERY AND RELATION TO APoE ALLELES G.D. Norata1,2 , K. Garlaschelli2 , L. Grigore2 , S. Raselli2 , S. Tramontana2 , G. Buccianti3 , A.L. Catapano1,2 . 1 Department of Pharmacological Sciences, University of Milan; 2 Centre for the Study of Atherosclerosis, Italian Society for the Study of Atherosclerosis, Bassini Hospital, Cinisello Balsamo; 3 The Italian Society for the Prevention of Kidney Disease, ASPREMARE, Milan, Italy E-mail: [email protected] Context: PCSK9 plays a key role in plasma cholesterol metabolism by modulating the expression of LDL receptors. Objective and Patients: In this study we investigated the effects of two common polymorphism of the PCSK9 gene (E670G and I474V) on the intima