4.P.412 LDL-apheresis in the treatment of familial hypercholesterolemia in childhood

Thursday 9 October 1997 : Posters Miscellaneous HDLC = 57 ± 16 (30-102) mg/dL) . Lipoproteins were separated in parallel with electrophoresis and standard methods, by sequential ultracentrifugation (Beckman TL-100). Levels of LDLC(e) were comparable with those of (u) (149 f 42 vs 148 ± 38 mg/dL, NS), whereas calculation underestimated LDLC levels by 15 % (135 ± 43 mg/dL, p < 10 -4 ) . Similarly, HDLC(e) levels were comparable with those observed after (p) (56 f 18 vs 57 ± 16 mg/dL, NS) whereas they both differed from HDLC(u) (44 ± 12 mg/dL, p < 10-4 ) . LDLC(e) significantly correlated with (u) (r2 = 0.87), and with (c) (r2 = 0.88) however patients with TG > 400 mg/dL were excluded for (c) . Therefore, this method allows routinely a direct and concomitant measure of LDL and HDLC at levels analogous with reference methods.

4 P.408

Correlation between plasminogen activator inhibitor and some lipids parameters

S . Kostovska, D . Lazove, M. Krstevska, S . Dzhekova-Stojkova, I . Dejanov, L . Lazarov. Institute of Blood Transfusion, Institute of Biochemistry, Clinic of CardiologyMedical Faculty, Skopje, Macedonia Plasminogen activator inhibitor (PAI-1 AG) is one of the most important components of the fibrinolytic systems since recent data suggest that is is a significant marker of thrombosis . The aim of the work was to establish the relations between PAM Ag and some lipids parameters (cholesterol-chol ., triglycerides-TG, HDL-chol, LDL-chol.) in patients with artherial diseases . The patients were divided into two groups: the first group consisted of 43 patients having acute myocardial infarction (AIM) while the second one counted 32 patients with endarteritis obliterans (EO) . PAI-Ag was determined by the immunoenzyme method (IMUBIND Plasma PAI-1 Elisa test, American diagnostica inc .) and the lipids with standard routine methods . The average values for PAI Ag were 77 ng/mL for AIM and 74 ng/mL for EO (n .v . = 5-52 ng/mL) . The following correlations were established . AIM : PAI/Cho1= 0.28 ; PAI/I'G = 0 .42 ; PAI/HDL = -0.22; PAI/LDL = 0 .18 ; EO:PAI = -44 ; 00PAI/TG = -0.31 ; PAI/HDL = 0 .01 and PAI/LDL = -0 .44 . Conclusion : The obtained results create a significant correlation between PAI-Ag and triglyceride levels, for the group with AIM . They also create a significant negative correlation between PAI-Ag and LDL and cholesterol, for EO, as well .

4 .P.409

Lipid disorders in patients with diabetes mellitus

D . Labudovik l , B . Todorova l , K. Petrovski 2 , T. Plasevski2 .'Dep . of Biochemistry, 2 Clinic of Endocrinology, Medical Faculty, Republik of Macedonia Patients with Type I (insulin-dependent) and Type II (non-insulin-dependent) diabetes mellitus are at increased risk of developing atherosclerotic vascular diseases . The lipid profile has been analyzed in fasted plasma taken from Type I (n = 40), Type II (n = 40) patients with DM, and from 55 healthy individuals (control group) . All lipid parameters were determined by standard biochemical diagnostic methods . Our investigation showed : (a) high statistical significant increase in the concentration of triglycerides, LDL-cholesterol and apolipoprotein B in both groups of patients with DM (p < 0 .01 ; p < 0 .001) compared to control group ; (b) the level of HDL-cholesterol was significant decreased in both groups of patients, and the decrease was more expressed in the patients with Type I DM (p < 0 .005) . As a conclusion it could be said that low HDL-Ch associated eith high TG,LDL-Ch which was proved with electrophoresis too, and high Apo B levels may be a strong factor for developing atherosclerotic vascular changes in patient with DM . The dislipidemia was more prominent in patients with Type I DM, so they have tendency to develop more expressed atherosclerotic vascular disease .

4. P.410

Competition-studies with autoantibodies to oxidized LDL

U . Resch, F. Tatzber, G . Waeg, H. Esterbauer. Institute of Biochemistry, Schubertstrafe 1, University of Graz, 8010 Graz, Austria In recent years, substantial evidence indicated that lipoproteins modified in vivo by radical-mediated oxidation or monenzymatic glycation may contribute to formation of autoantibodies to oxidized LDL in normal subjects and in patients with atherosclerosis . Contradictory data reported by different groups make it difficult to associate the presence or progression of atherosclerotic

383

lesions with the occurrence of autoantibodies . We tried to eludicate the type of antigen that promotes the formation of antibodies recognizing oxidized or modified LDL. In a first step, we screened subjects in a population of Graz (about 200 subjects, clinically healthy and atherosclerotic patients) to find high autoantibody titers by using Cu 2+ -oxidized LDL as antigen. As mentioned above, the antigen which causes the formation of these antibodies is not known and it cannot be excluded that the antigen is an oxidized protein other than apoB . In a second step, an individual serum with high antibody titer was examined by competitive ELISAs using a wide range of competitors . We examined LDL modified by various lipid peroxidation products, such as HNE or MDA, macrophage-modified and enzyme (lipozygenase)-modified LDL, oxidized HLD erythrocytes and oxidized microsomes . With this individual serum, oxidized L!DL was the best competitor but other antigens like lipoxygenasemacrophage- or MDA-modified LDL and oxidized erythrocytes also competed with the binding of the antibodies to the solid phase . In conclusion, it seems that autoantibodies to oxidized LDL do not predominantly recognize oxidized LDL, but also modified LDL's and proteins . They show variable cross-reactivity and have moderate-to-low affinity . The results suggest that, once an antibody is generated against an adduct between a lipid oxidation product and the apoprotein, it could also react with similar adducts formed elsewhere in the body . It may be of potential clinical relevance to detect the presence of autoantibodies . This can be used as an indicator of the extent of atherosclerosis or degree of in vivo oxidation (Austria Science Foundations S 7102-MED) .

4.P.411

Diet only and diet plus simvastatin in the treatment of heterozygous familial hypercholesterolemia in childhood .

C . Stefanulti, S . Di Giacomo, A . Vivenzio, G . Bosco', V. Colloridil, A . Bucci. Plasmapheresis Unit, Istituto di Terapia Medica Sistematica, 'Pediatric Cardiology Service, Istituto di Clinica Pediatrica, Policlinico Umberto I. University Of Rome "La Sapienza ", Rome, Italy One year clinical study on No . 16 (No. 7 males and No . 9 female) pediatric patients with heterozygous familial hypercholesterolemia treated with hypocholesterolemic diet only or with diet plus drug (simvastatin), was carried out . According to the study protocol, the children were submitted to a three months wash out free diet ; then they were given a diet (step II AHA) for six months . After six months, they were divided into two groups matched for sex, age and BMI. Diet only was given to the group A (No . 8) ; simvastatin (10 mg/daily) was given to the group B, for one year respectively . All patients were examined at baseline, and monitored during the study by pediatricians . All patients were submitted to non invasive CVD examination (exercise ECG, echocardiography) . We failed to observe side effects during the treatment with diet, and, with diet plus simvastatin, on relatively long-term treatment . The table shows mean plasma Total Cholesterol (T-CHOL), Triglyceride (TG), HDI: Cholesterol (HDL-C) and LDL-cholesterol (LDL-C) levels, during six months of diet administration only, and after one year in both groups . All patients No . 16 Months T-CHOL HDL-C LDL-C TG

0 280(21) 56(6) 201 (41) 121 (30) (Step II AHA

6 268(27) 54(5) 195(32) 106(28) diet)

Group A No . 8 0

Group B No. 8 12

211 (37) 256(41) 54(9) 50 (12) 149(67) 142(25) 105(46) 89(32) (Step IIAHA diet)

0

12

331 (69) 251 (58)' 51 (9) 55(12) 256(71) 180(87)' 95(36) 79(26) Simvastatin 10 and/d)

t mg/dl; * po1 .01 .

After twelve months of treatment with simvastatin T CHOL, and LDL-C showed a significant reduction . The increase of T CHOL, and LDL-C in patients or, diet only, was of 4 and 3% and 17 and 4% after 6, and 12 months, respectively.

4 .P.412' LDL-apheresis in the treatment of familial hypercholesterolemia in childhood C . Stefanutti, S . Di Giacomo, A . Vivenzio, G . Bosco', V. Colloridi, S. Trizza, A . Berni 2 , A. Nigri 2 , N . Koga3 . Plasmapheresis Unit, Istituto Di Terapia Medico Sistematica, 'Pediatric Cardiology Unit, Istituto Di Clinica Pediatrica, 2 Haemodinamic Unit I, Istituto di Chirurgia Del Coure e Dei Grossi Vasi, Policlinico Umberto L, University of Rome "La Sapienza ", Rome, Italy; 2 Department of Cardiology & Artificial Organs, Koga Hospital, Kurume City, Japan We treated 8 children (aged 4 .5-13 years, No . 2 males, No . 6 females) with homozygous and double heterozygous familial hypercholesterolemia (HFH,

11th International Symposium on Atherosclerosis, Paris, October 1997



Thursday 9 October 1997: Posters Miscellaneous

384

DHFH) with LDL-apheresis (LDL/A), using three different selective and semiselective techniques : dextransulphate cellulose (DSC-LDL/A) (Liposorber MA-01, Kancka Corp. Osaka Japan) . Heparin mediated Extracorporeal LDL Precipitation (HELP-LDL/A) (Plasmat, SECURA, B . Braun, Melsungen, Germany), diacetate cellulose cascade filtration (DF-LDL/A) (BT 985, DIDECO, Mirandola-Modena, Italy) and conventional plasmaexchange (PE) (BT 798 CE, DIDECO, Mirandola-Modena, Italy) . The patients were submitted to a comprehensive program of genetic, molecular, clinical and metabolic study, alone with invasive and non invasive cardiovascular examinations . The youngest (4 .5 years) patient with HFH ever been treated with LDL/A was in this sample. The interval between treatments was of fifteen days . All patients, with the exception of one, were affected by coronary artery disease . Until now, we performed n . 361 LDL-aphereses, and PE in pediatric patients . We failed to observe major severe side effects during treatment . In fact, all procedures were regularly completed. Two patients with symptomatic angina, at entry, and with three vessels, extremely severe coronary artery disease, demonstrated by coronary angiography, died after four complete LDL/aphereses . The other children, still on treatment, are growing up showing progressive metabolic improvement, regression of skin xanthomas . Two of them, showed complete absence of progression and regression of coronary atherosclerotic lesions, demonstrated by serial coronary angiographics . The table shows mean plasma LDL cholestorol (LDLC) apolipoprotein B (APO B) and L(a) levels in one of the above mentioned patients who was submitted to four different apheretical techniques : Help LDLC' APO B LP(a)

429 266 53

DSC 151 120 17

443 281 54

DF 98 95 18

471 302 51

PE 111 102 20

391 261 49

62 90 19

mg/dl .

We submitted to LDL/A the yougest patient ever been treated by means of LDL/A, as far as we know from the international literature . In our hands, LDL/A showed to be a useful and succesful tool to be used even in the treatment of severe hypercholesterolemic pediatric patients . Our experience seems to strongly suggest a premature use of LDL/A in the treatment of severe metabolic impairment in childhood .

4 .R413

Effect of fluvastatin on serum lipoproteins and glycemic control in hyperlipidemic patients with type II diabetes

A . Torn, A . Branchi l , A. Rovellini l , S . Gasperini, D . Sommariva . Department of Internal Medicine, Hospital of Bollate ; 'Institute of Internal Medicine and Medical Physiopathology, University of Milan, Ospedale Maggiore IRCCS, Milan, Italy Type II diabetic patients frequently have higher than normal serum lipids in spite of a good metabolic control . In order to reduce cardiovascular risk, it may be advisable to treat the patients with hypolipidemic drugs which do not adversely affect glucose metabolism . Fifteen hypercholesterolemic type II diabetic patients in stable glycemic control have been treated with fluvastatin 20 mg once a day. After 2 months serum cholesterol fell by 21% and LDL cholesterol by 32% . HDL cholesterol and serum triglycerides did not change significantly . Fasting blood sugar and glycated hemoglobin did not change during the treatment . Totall cholesterol LDL cholesterol HDL cholesterol Serum triglycerides Fasting blood sugar HbAlc

Basal

2 months

P

299.5 t 11 .16 192 .2 t 8 .61 50.0 f 4.58 286.5 t 41 .55 151 .9 f 10.78 651±0.39

237.7 ± 7.41 130.7 ± 7.50 50 .9 t 3 .25 280 .8 ± 48 .09 157 .0 ± 10 .43 6 .61 ± 0 .42

< 0.001 < 0.001 N.S . N .S . N .S . N .S .

One patient stopped the treatment after 2 months because of heartburn . The continuation of the therapy for an additional 3 months in 9 patients did not produce further changes in total and LDL cholesterol . Glycated hemoglobin remained similar to the pretreatment level . Fluvastatin is therefore effective in hyperlipidemic diabetic patients in lowering serum cholesterol without adversely affecting glucose control.

cholesterol, triglycerides, high-density lipoprotein cholesterol, and apolipoproteins A-I and B in cord blood, from children bom in our hospital for a five-month period . Cholesterol and triglycerides concentrations have been measured by enzymatic full automized methods . Apolipoproteins A-I and B, and lipoprotein (a) levels have been obtained by rate immunonephelometric assay. HDL lipoproteins have been separated by precipitation with polyethylenglycol . Cholesterol and triglycerides values have been lightly lower than others values obtained in cord blood in other populations . The distribution of lipoprotein (a) concentrations has been markedly skewed to low values (mean = 1 .74 mg/dl ; median = 1 mg/dl), similar to that other young or adult Caucasian populations . As compared with children with a negative familial history of cardiovascular heart disease, those with a positive history have had a higher mean lipoprotein (a), and a higher prevalence of lipoprotein values exceeding 5 mg/dl . These results suggest that lipoprotein (a) is an important marker of risk, from birth, for cardiovascular heart disease .

4 .P.415

J .C. Vella, P. Calmarza l , L . Castillo', A . Giner2 . Junta de Castilla y Leon, B urgos ; 'Hospital G . Yagiie, Burgos ; 2 Hospital C. Universitario, Zaragoza, Spain The aim of this study has been to know the concentration of lipoprotein (a), and to determine the distribution of apolipoprotein (a) phenotypes in non-insulin dependent diabetic subjects . Lipoprotein (a ) concentrations have been measured by rate immunonephelometric assay, and apolipoprotein (a) phenotypes by electrophoresis and immunoblotting in 107 non-insulin dependent patients . The distribution of lipoprotein (a) concentrations has been markedly skewed, with the highest frequencies at low values, and 21 mg/dl as mean value, and 14 mg/dI as median value . The more prevalent phenotypes have been S2 (25%), SO (24%) and S3 (20%) . The subjects with phenotypes containing F allele, have shown the highest lipoprotein (a) concentrations : 85 mg/dl for the homozygous state, and 48 mg/dl for the heterozygous state . Lipoprotein (a) concentrations and apolipoprotein (a) phenotypes distribution in our patients, have been similar to those observed in healthy subjects, so the increased risk for atherosclerosis in non-insulin dependent diabetic subjects, cannot be attributed to neither the lipoprotein (a) concentrations nor the apolipoprotein (a) phenotypes . The F homozygous state has shown higher concentrations of both LDLcholesterol and apolipoprotein B, than average values .

I 4 .P.416 Butter, margarine and lipoprotein cholesterol in dietary trails P.L . Zock, M.B . Katan . Dept . of Human Nutrition, Wageningen Agricultural University, Bomenweg 2, 6703 HD Wageningen, The Netherlands Margarines have long had a "healthy" image because they are lower in saturated fatty acids and cholesterol than butter . However, margarines are also a major source of trans fatty acids, which have recently been found to unfavorably affect serum lipoproteins . Therefore, the advantages of margarines over butter have become controversial . We conducted a meta-analysis of 20 trials that directly compared the effect of butter and margarine on blood lipids in humans . Replacing 10% of calories from butter with hard stick margarine lowered total serum cholesterol by 0.19 mmol/L but did not affect the total/HDL cholesterol ratio (4 trials, 172 subjects) . Soft tub margarines lowered total cholesterol by 0 .25 mmol/L and the total/HDL ratio by 0 .20 (8 trials, 287 subjects) . These findings can be explained by the absolute amounts of trans and saturated fatty acids in butter and margarines . Based on total/HDL cholesterol ratio, we estimate that replacing 10% of calories or 30 g of butter with soft low-trans margarines reduces coronary heart disease risk by 10%, whereas hard high-trans margarines are ineffective .

4 .P.417 4 .P414

Lipoprotein (a) and other lipidic parameters in newborns from Burgos, Spain

Lipoprotein (a) and apolipoprotein (a) phenotypes in non-insulin dependent diabetic subjects

Cholesterol ester fatty acids as quantitative indicators of fatty acid intake in humans

J .C . Vella, P. Calmarza l , M. Goili l , J . Berzosa l . Junta de Castilla y Leon, 'Hospital G . Yagiie, Burgos, Spain

P.L . Zock l , R .P. Mensink 2 , J . Harryvan l , J .M .H . de Vries l , M .B . Katan l . 'Department of Human Nutrition, Wageningen Agricultural University, Wageningen, The Netherlands ; 2 Department of Human Biology, Limburg University, Maastricht, The Netherlands

The purpose of this study has been the concentrations of lipoprotein (a), total

The fatty acid composition of serum cholesteryl esters is used as a qualitative

11th International Symposium on Atherosclerosis, Paris, October 1997