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1.P.392 The influence of apolipoprotein E isoforms in metabolic abnormalities and sodium-lithium countertransport activity of hypertensive patients
Monday
6 October 1997: Posters Methods
loo the existence of an important adaptative process in the maintenance tone during pathological situations (i.e. aging or atherosclerosis) with an impaired endothelium-dependent vasorelaxation.
El1 P.391
METHODS
of vascular associated
m1
of corvaton and renitw in unstable angina patients with hypertension: Influence on platelet-endothelial interactions
The use
A.A. Semvonkin, Internal Diseases,
V.M. Jakovlev, Omsk Medical
P.V. Jakovlev, G.S. Sokolov. Academy, Omsk Russia
Department
of
The aim of the study was to evaluate the influence of simultaneous administration of Corvaton [C] and Renitec [R] on the course of acute coronary syndrome in unstable angina (UA) patients with hypertension. The study included 40 hypertensive patients with clinic of UA (Braunwald’s class Ba, IIb). All patients were randomly assigned either to C (8-16 mg/day) and R (10-20 mg/day) or to Isosorbid Dir&ate (60-120 mg/day) and R (10-20 mg/day), Group 1 (n = 22) and Group 2 (n = 18), respectively. Conventional methods of clinical investigation were accomplished by evaluation of platelet activity and von Villebrand factor by modified Born’s method. The table shows changes of platelet-endothelial function in both groups. Gmup 1
Spontaneous aggr. ADP induced (IO wM) Collagen induced (2.5 &ml) YO” Vdlebrand factor
P
Group 2
admission
8th day
admission
8th day
2.8 ck 0.3 76.5 zt 12.3 82.3 z!z 10.3 145.6 i 21.3
1.3 *0.1** 48.2 f6.1** 42.1 zk 7.2** 96.2 f7.8*'
3.0 * 0.4 78.3 z!z14.5 84.1 i 12.0 150.2 * 24.2
2.6 i OS’ 69.4 i 11.7 68.3 f9.8' 12S.Ozk 10.6*
Decrease of platelet activity and von Villebrand factor was noted in both groups, however, significant changes were observed only in group 1. Blood pressure was comparable in the two groups, but clinical outcomes were different. MI developed in 1 patient (4.5%) in Group 1 versus 3 patients (16.6%) in group 2. Significantly lower number of incidences of arrhythmias and anginal episodes occurred in group 1. There were no angina at rest episodes and the need for Aspirin or Heparin appeared in rare cases in this group. Thus, additive effect of C and R on platelet-endothelial interactions leads to better prognosis in hypertensive patients with UA. The use of C is more preferable than nitrates in this situation.
Tbe influence of apolipoproteti E isoforms in metabolic abnormalities and sodium-lithium countertransport activity of hypertensive patients
References [1] Tato F., Arterioscler.
Introduction:
To determine the influence of ApoE polymorphism on plasma lipoprotein profile, erythrocyte Na+/Li+ countertransport, carbohydrate and insulin metabolism in patients with untreated essential hypertension. Subjects and Methods: 84 patients were consecutively enrolled for the present study, subdivided into three groups carrying E2, E3 or E4 allele.
Results: ApoE (n = 9)
ApoE (n = 61)
ApoE (n = 14)
56.0 * 10.5 29.1 f 3.9 229.1 f 35.1 140.9 * 20.5 51.8 f 12.4 2.7 zt 0.7 139.1 i51.7 163.2~k32.9~ 109.8 f 13.5 40.1 i 2.8++ 0.60 * 0.14 0.327 ztO.09 414f414
47.9 5 10.8 29.1 f 3.8 225.5 f54.5 144.5 + 46.5 48.1 zk 12.5 3.2 f 1.3 158.8 z!z 100.6 133.8 i27.0 112.7 f 33.2 46.2 i 6.8 0.87 + 0.25 0.365 10.13 912f469++
47.1 f 10.1 28.8 zk 4.5 256.3 f 49.9 169.2 i49.3 42.3 i 9.4 4.1 * l.l++ 205.9 zk 90.4 135.3 z!c 18.2 138.0 f 32.8+ 45.7 f 5.5 0.86 + 0.21 0.354 * 0.11 631f490
Con&sions: Our data suggest that apoE polymorphism metabolic abnormalities and Na+/Li+ countertransport activity essential hypertension.
11th International
Vast. Biol.
1997, 17: 56-63.
Respiratory activity of rabbits liver mitochondria in experimental atherosclerosis
In regard to the central role of the liver in lipid metabolism, we examined the function of liver mitochondria in experimental atherosclerosis. Experimental atherosclerosis in rabbits was provoked with a 2% cholesterol diet. We fed New Zeland White rabbits during 12 weeks by either 2% cholesterol (8 mg/kg) (E, n = 14) or 154 mmol/L NaCl (K, n = 18). We prepared liver mitochondria using Chapel1 method. Mitochondrial function was tested by measuring by measuring oxygen consumption (State 3 (+ADP)) with oxygen electrode of the Clark type. The result showed that oxygen consumption of liver mitochondria in the control group was 1.808 f 0.44 ng atom O/min/mg proteins and in the experimental group 0.828 f 0.21 ng atom O/min/mg proteins. Oxygen consumption was reduced by 54.18% (p < 0.05). Respiratory activity of rabbits mitochondria in experimental atherosclerosis was reduced.
crl1 P.395
L. Beghin, Department Pharmacie,
may modulate in patients with
Symposium
394
Thromb.
1.2. Andielkovic, D.D. Andjelkovic, Z.Z Kojic, D.M. Djuric, M.M. RosiC. Department of Physiology, University School of Medicine, Belgrade, Serbia
C. Montilla, 0. Mu&z-Grijalvo, P. Stiefel, A. Alonso, M. Gotor, A. Femandez-Moyano, R. Garcia-Lozano, A. Nuiiez, J. Villar. Hypertension and Lipids Unit, Inmunological Service, Hospital Universitario Virgen de1 Rocio, &villa, Spain
Age (years) BMI (kg/m*) Total Cholesterol (mg/dl) LDLc (mg/dl) HDLc (mg/dl) LDLc/HDLc Tryglicerides (mg/dl) Apnlipopmtein Al Apolipopmtein B Membrane Cholesterol (%) ChoWh membrane ratio NatiLi+countemanspat Peptide Gm uncicrcurve
Fluorescent determination of CETP activity in plasma
The determination of plasma CETP activity with isotopically labeled lipoproteins is known to be laborious. We report a method to assess CETP activity in a defined system by using flourescent cholesteryl linoleate analog in comparison to an isotopic assay (1). Method: Exogenous donor (MW 756) and acceptor particles (human VLDL) are combined with CETP source (plasma) within a total volume of a 10 mM tris buffer (pH 7.4). Donor particles consist of a nitrobenzoxadiol-fluorophor which takes advantage of a self-quenching effect on microemulsion incorporated cholesteryl linoleate (WAK, Germany). CETP mediated transfer is determined by the increase of fluorescence intensity as the cholesteryl linoleate is removed from the self quenched donor to the acceptor particle (incubation tune 3 h at 37” C; EX 435 nm, EM 535 using a LS.50 fluorescence photometer (Perkin Elmer)). A standard curve to derive the relation between fluorescence intensity and mass transfer is generated from a linear dilution of a defined concentration of the donor particle in isopropanol. Results: After screening of 50 plasma samples, a nearly 10 fold individual transfer rate between the lowest (6 pmoV3 h) and the highest (57 pmoU3 h) one was observed. 10 series of 10 samples and a series of 20 measures of 3 samples in different CET activities (15/34/46 pmol/3 h) show an interassay CV of 14.6%, and an intraassay CV of 10.8% (all samples were measured in duplicates). A good correlation is found between CET rates of 20 samples obtained from the fluorescent assay (range 5-65 pmoV3 h) and from the isotopic assay (r = 0.9, p = 0.000). Conclusion: Taken together, this new fluorimetric assay for determining CETP activity has the advantage of avoiding radioactive substrate. In addition, it provides a fast, simple and specific method for quantifying CETP activity in the clinical laboratory.
I1 P 1 P.392 LIZI
393
A. Ambrosch, F. Tato’, C. Luley. Institutfir Klinische Chemie, Uniklinik Magdeburg; ‘Medizinische Poliklinik, Ludwig-Maximillians-Universitat Miinchen, Germany
Measurements of VLDL-, IDL-, and LDL-apo B levels by a combination of isopropanol precipitation and mass spcetrometry N. Duhal, B. Lacroix, J.P. Bonte, J.C. Fruchart, G. Luc. OfAtherosclerosis, Pasteur Institute of Lille, Faculte’ de Lille, France
The determination of plasma apolipoprotein B- 100 level, the major protein component of apo B-100 containing lipoproteins is usually carried out by immunological assays. However, these methods arc subject to criticism as regards the quantifying of apo B-100 in purified lipoproteins. Chemical methods have therefore been developed, such as the use of tetmmethylurea or isopropanol to precipitate apo B selectively. A method is presented here for the quantifying of apolipoprotein B-100 concentration in lipoproteins isolated by ultracentrifugation and by a combination of isopropanol precipitation and gas chromatography-mass spectrometry. The apo B-100 is precipitated by isopropanol and quantitated by a mass
on Atherosclerosis,
Paris,
October
1997
6 October 1997: Posters Methods
loo the existence of an important adaptative process in the maintenance tone during pathological situations (i.e. aging or atherosclerosis) with an impaired endothelium-dependent vasorelaxation.
El1 P.391
METHODS
of vascular associated
m1
of corvaton and renitw in unstable angina patients with hypertension: Influence on platelet-endothelial interactions
The use
A.A. Semvonkin, Internal Diseases,
V.M. Jakovlev, Omsk Medical
P.V. Jakovlev, G.S. Sokolov. Academy, Omsk Russia
Department
of
The aim of the study was to evaluate the influence of simultaneous administration of Corvaton [C] and Renitec [R] on the course of acute coronary syndrome in unstable angina (UA) patients with hypertension. The study included 40 hypertensive patients with clinic of UA (Braunwald’s class Ba, IIb). All patients were randomly assigned either to C (8-16 mg/day) and R (10-20 mg/day) or to Isosorbid Dir&ate (60-120 mg/day) and R (10-20 mg/day), Group 1 (n = 22) and Group 2 (n = 18), respectively. Conventional methods of clinical investigation were accomplished by evaluation of platelet activity and von Villebrand factor by modified Born’s method. The table shows changes of platelet-endothelial function in both groups. Gmup 1
Spontaneous aggr. ADP induced (IO wM) Collagen induced (2.5 &ml) YO” Vdlebrand factor
P
Group 2
admission
8th day
admission
8th day
2.8 ck 0.3 76.5 zt 12.3 82.3 z!z 10.3 145.6 i 21.3
1.3 *0.1** 48.2 f6.1** 42.1 zk 7.2** 96.2 f7.8*'
3.0 * 0.4 78.3 z!z14.5 84.1 i 12.0 150.2 * 24.2
2.6 i OS’ 69.4 i 11.7 68.3 f9.8' 12S.Ozk 10.6*
Decrease of platelet activity and von Villebrand factor was noted in both groups, however, significant changes were observed only in group 1. Blood pressure was comparable in the two groups, but clinical outcomes were different. MI developed in 1 patient (4.5%) in Group 1 versus 3 patients (16.6%) in group 2. Significantly lower number of incidences of arrhythmias and anginal episodes occurred in group 1. There were no angina at rest episodes and the need for Aspirin or Heparin appeared in rare cases in this group. Thus, additive effect of C and R on platelet-endothelial interactions leads to better prognosis in hypertensive patients with UA. The use of C is more preferable than nitrates in this situation.
Tbe influence of apolipoproteti E isoforms in metabolic abnormalities and sodium-lithium countertransport activity of hypertensive patients
References [1] Tato F., Arterioscler.
Introduction:
To determine the influence of ApoE polymorphism on plasma lipoprotein profile, erythrocyte Na+/Li+ countertransport, carbohydrate and insulin metabolism in patients with untreated essential hypertension. Subjects and Methods: 84 patients were consecutively enrolled for the present study, subdivided into three groups carrying E2, E3 or E4 allele.
Results: ApoE (n = 9)
ApoE (n = 61)
ApoE (n = 14)
56.0 * 10.5 29.1 f 3.9 229.1 f 35.1 140.9 * 20.5 51.8 f 12.4 2.7 zt 0.7 139.1 i51.7 163.2~k32.9~ 109.8 f 13.5 40.1 i 2.8++ 0.60 * 0.14 0.327 ztO.09 414f414
47.9 5 10.8 29.1 f 3.8 225.5 f54.5 144.5 + 46.5 48.1 zk 12.5 3.2 f 1.3 158.8 z!z 100.6 133.8 i27.0 112.7 f 33.2 46.2 i 6.8 0.87 + 0.25 0.365 10.13 912f469++
47.1 f 10.1 28.8 zk 4.5 256.3 f 49.9 169.2 i49.3 42.3 i 9.4 4.1 * l.l++ 205.9 zk 90.4 135.3 z!c 18.2 138.0 f 32.8+ 45.7 f 5.5 0.86 + 0.21 0.354 * 0.11 631f490
Con&sions: Our data suggest that apoE polymorphism metabolic abnormalities and Na+/Li+ countertransport activity essential hypertension.
11th International
Vast. Biol.
1997, 17: 56-63.
Respiratory activity of rabbits liver mitochondria in experimental atherosclerosis
In regard to the central role of the liver in lipid metabolism, we examined the function of liver mitochondria in experimental atherosclerosis. Experimental atherosclerosis in rabbits was provoked with a 2% cholesterol diet. We fed New Zeland White rabbits during 12 weeks by either 2% cholesterol (8 mg/kg) (E, n = 14) or 154 mmol/L NaCl (K, n = 18). We prepared liver mitochondria using Chapel1 method. Mitochondrial function was tested by measuring by measuring oxygen consumption (State 3 (+ADP)) with oxygen electrode of the Clark type. The result showed that oxygen consumption of liver mitochondria in the control group was 1.808 f 0.44 ng atom O/min/mg proteins and in the experimental group 0.828 f 0.21 ng atom O/min/mg proteins. Oxygen consumption was reduced by 54.18% (p < 0.05). Respiratory activity of rabbits mitochondria in experimental atherosclerosis was reduced.
crl1 P.395
L. Beghin, Department Pharmacie,
may modulate in patients with
Symposium
394
Thromb.
1.2. Andielkovic, D.D. Andjelkovic, Z.Z Kojic, D.M. Djuric, M.M. RosiC. Department of Physiology, University School of Medicine, Belgrade, Serbia
C. Montilla, 0. Mu&z-Grijalvo, P. Stiefel, A. Alonso, M. Gotor, A. Femandez-Moyano, R. Garcia-Lozano, A. Nuiiez, J. Villar. Hypertension and Lipids Unit, Inmunological Service, Hospital Universitario Virgen de1 Rocio, &villa, Spain
Age (years) BMI (kg/m*) Total Cholesterol (mg/dl) LDLc (mg/dl) HDLc (mg/dl) LDLc/HDLc Tryglicerides (mg/dl) Apnlipopmtein Al Apolipopmtein B Membrane Cholesterol (%) ChoWh membrane ratio NatiLi+countemanspat Peptide Gm uncicrcurve
Fluorescent determination of CETP activity in plasma
The determination of plasma CETP activity with isotopically labeled lipoproteins is known to be laborious. We report a method to assess CETP activity in a defined system by using flourescent cholesteryl linoleate analog in comparison to an isotopic assay (1). Method: Exogenous donor (MW 756) and acceptor particles (human VLDL) are combined with CETP source (plasma) within a total volume of a 10 mM tris buffer (pH 7.4). Donor particles consist of a nitrobenzoxadiol-fluorophor which takes advantage of a self-quenching effect on microemulsion incorporated cholesteryl linoleate (WAK, Germany). CETP mediated transfer is determined by the increase of fluorescence intensity as the cholesteryl linoleate is removed from the self quenched donor to the acceptor particle (incubation tune 3 h at 37” C; EX 435 nm, EM 535 using a LS.50 fluorescence photometer (Perkin Elmer)). A standard curve to derive the relation between fluorescence intensity and mass transfer is generated from a linear dilution of a defined concentration of the donor particle in isopropanol. Results: After screening of 50 plasma samples, a nearly 10 fold individual transfer rate between the lowest (6 pmoV3 h) and the highest (57 pmoU3 h) one was observed. 10 series of 10 samples and a series of 20 measures of 3 samples in different CET activities (15/34/46 pmol/3 h) show an interassay CV of 14.6%, and an intraassay CV of 10.8% (all samples were measured in duplicates). A good correlation is found between CET rates of 20 samples obtained from the fluorescent assay (range 5-65 pmoV3 h) and from the isotopic assay (r = 0.9, p = 0.000). Conclusion: Taken together, this new fluorimetric assay for determining CETP activity has the advantage of avoiding radioactive substrate. In addition, it provides a fast, simple and specific method for quantifying CETP activity in the clinical laboratory.
I1 P 1 P.392 LIZI
393
A. Ambrosch, F. Tato’, C. Luley. Institutfir Klinische Chemie, Uniklinik Magdeburg; ‘Medizinische Poliklinik, Ludwig-Maximillians-Universitat Miinchen, Germany
Measurements of VLDL-, IDL-, and LDL-apo B levels by a combination of isopropanol precipitation and mass spcetrometry N. Duhal, B. Lacroix, J.P. Bonte, J.C. Fruchart, G. Luc. OfAtherosclerosis, Pasteur Institute of Lille, Faculte’ de Lille, France
The determination of plasma apolipoprotein B- 100 level, the major protein component of apo B-100 containing lipoproteins is usually carried out by immunological assays. However, these methods arc subject to criticism as regards the quantifying of apo B-100 in purified lipoproteins. Chemical methods have therefore been developed, such as the use of tetmmethylurea or isopropanol to precipitate apo B selectively. A method is presented here for the quantifying of apolipoprotein B-100 concentration in lipoproteins isolated by ultracentrifugation and by a combination of isopropanol precipitation and gas chromatography-mass spectrometry. The apo B-100 is precipitated by isopropanol and quantitated by a mass
on Atherosclerosis,
Paris,
October
1997