- Email: [email protected]
Establishment of reference range for hdl subfractions in japanese population with a new automated assay
Abstracts / Atherosclerosis 235 (2014) e84–e191
E. Shakhtshneidera, Y. Raginob, Y. Polonskayab, E. Kashtanovab, Y. Nikitinc, S. Malyutinac, M. Voevodaa a Laboratory of Genetics of Internal Medicine, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia; b Laboratory of Biochemistry, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia; c Laboratory of Internal Medicine, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia
Objectives: We have analyzed the frequencies of HindIII polymorphism of lipoprotein lipase gene (LPL) and lipid profile in Caucasian population of West Siberia and the groups this high and low total cholesterol (TC) level. Methods: The patients included in the analyses were selected based on TC level from population sample surveyed in frame of HAPIEE project (w9000 participants, aged 45-69, men 50%). Totally 259 patients with highest TC level (>300mg/dl) and 228 patients with lowest TC level (<200mg/dl) and 170 randomly selected patients (mean TC level - 235.843.9mg/dl) were included. The differences of TC level between groups are significant. The plasma lipid levels were determined by standard enzymatic assays. The subfractional profile of LDL was determined by method of electrophoresis. The HindIII polymorphism (22125T/G) of LPL gene was analyzed by RELFPCR. Results: Frequencies of H+H+, H+H- and H-H- genotypes were 61%, 35% and 4% in population. The frequency of H- allele was 0.22, 0.28 and 0.29 in HAPIEE population, low and highest TC level groups, respectively (p>0.05). We have found the association of HindIII polymorphism of LPL gene with TG level in HAPIEE population (p¼0.01). We have found the association of polymorphism HindIII of gene LPL with minor peak on a descending curve of subfractional profile LDL (p¼0,02). Conclusion: The Caucasian population of West Siberia is not significantly differs from populations of Europe by frequencies of alleles and genotypes. The genotype H+H+ of HindIII polymorphism of LPL gene has been associated with high TG level. The reported study was partially supported by RFBR, research project N1404-01594a. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0119. HDL AS A BIOMARKER OF REJECTED HEART PRODUCING SEVERE ATHEROSCLEROSIS OF THE TRANSPLANTED HEART W. Gonzáleza, P. Altieria, H.L. Banchsa, D. Iravedraa, I.F. González-Cancelb, R. Calderóna, C. Gurreab, N. Escobalesa, M. Crespoa a Medicine and Physiology, University of Puerto Rico Medical Sciences Campus, San Juan, Puerto Rico; b Heart Transplant Program, Cardiovascular Center of Puerto Rico and the Caribbean, San Juan, Puerto Rico
Objectives: To study the metabolic HDL changes post heart transplant. Methods: 101 Patients (P.) with heart transplant were studied. 65 P. were males and 36 were female, with a mean age of 51 years. Results: Pre and post metabolic changes were: 1. BMI (25 4 – 28 Kg/m2 P < 0.05). 2. Systolic blood pressure (107 13 – 131 20 P <0.05). 3. Diastolic blood pressure (70 13 – 81 10 mmHg). 4. Fasting blood sugar (107 37 – 117 55 mg%) N.S. 5. Total cholesterol (170 55 – 189 32mg/dl) P < 0.05. 6. Total HDL (38 16 – 52 17mg/dl) P < 0.05. 7. Total LDL (99 20 – 83 15mg/dl) N.S. 8. Triglycerides (163 10 – 188 12mg/dl) N.S. Conclusion: Our data shows that all the P. developed Mets. The HDL of the whole groups increased from 38 mg/dl to 52 mg/dl. These changes are not explained only by the immunosuppressive treatment, HDL has a role as an anti-inflammatory agent which is inherent in the lipoprotein characteristics. 16 P. died of rejection. The HDL of the rejected P. increased from 47 22 to 71 40 mg/dl P. < 0.007. This increase persisted through the course
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of the rejection period. The mean age of the transplanted heart was 22 years. The rejection occurred in less than 5 years. (3.5 years). Seven autopsies were done. Five of them showed severe atherosclerotic changes in the coronaries and aorta. This is probably explained on basis of dysfunctional HDL. Probably the high levels and persistent elevation of HDL in the rejected group can be used as a biomarker to predict rejection. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0807. HDL, ITS SUBFRACTIONS AND APOLIPOPROTEIN-A1 AS PREDICTORS OF HIGH PLATELET REACTIVITY AND MAJOR ADVERSE CARDIOVASCULAR EVENTS IN PATIENTS WITH STEMI R. Grzybczak, A. Siniarski, J. Zalewski, J. Nessler, G. Gajos Department of Coronary Disease John Paul II Hospital, Jagiellonian University Medical College, Krakow, Poland Objectives: Determine the relationship between serum HDL-cholesterol levels and its subfractions (LpA-I and LpA-I/A-II) and apoprotein A1 (Apo A-I) and early high platelet reactivity (HPR) on clopidogrel in the acute phase of STEMI. Methods: Fifty-two patients (14 women and 38 men) aged 60.6 9.14 years old with STEMI were included in this study. Patients (pts) were treated with primary PCI with stent implantation and optimal medical therapy including aspirin and clopidogrel for 12 months. After 12 hours of the STEMI onset we assessed: HDL and its subfractions (enzymatic and rocket immunoelectrophoresis, respectively); platelet reactivity (optical aggregometry and flow cytofluorometry). The study group was divided into 2 subgroups: with high and low levels of HDL-C, Apo A-I, LpA-I and LpA-I/A-II according to post-hoc analysis of the median concentration. During 12-months followup we assessed the incidence of major adverse cardiovascular events. Results: Higher values of LpA-I and lower concentrations of LpA-I/A-II were associated with HPR despite clopidogrel treatment. LpA-I levels higher than 57.52 mg/dl predicted HPR. The risk of inadequate response to antiplatelet therapy increased by 3% with every 1 mg/dl increase of LpA-I level. Moreover, HDL-C and Apo A-I over 1.19 mmol/l and 129.6 mg/dl respectively were significant predictors of rehospitalization in 12-month followup. The cut-off point 4 mg/dl for D12 LpA-I/A-II by the ROC curve analysis most accurately predicted rehospitalization with 66% specificity and 100% sensitivity. Moreover, LpA-I over 58.27 mg/dl was associated with higher risk of unplanned coronary revascularization. Conclusion: Increased LpA-I concentration predicted HPR despite antiplatelet treatment in pts with acute STEMI. Higher LpA-I levels indicated also increased risk of major cardiovascular events. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0631. ESTABLISHMENT OF REFERENCE RANGE FOR HDL SUBFRACTIONS IN JAPANESE POPULATION WITH A NEW AUTOMATED ASSAY Y. Ito, N. Satoh, T. Ishii, J. Kumakura Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan Objectives: Measurement of high-density lipoprotein (HDL) 2 and HDL3 subfractions might be more useful for evaluating coronary risk than total HDL-cholesterol (C). However, methods of measuring HDL2 and HDL3 are quite laborious for general clinical use and thus so far it has been difficult to establish reference ranges. We carried out a study to establish a reference range for HDL subfractions in Japanese population using our novel homogeneous assay for HDL subfractions. Methods: HDL-C and HDL3-C were measured by our homogeneous assays on a Hitachi 917 automated clinical chemistry analyzer (Hitachi). HDL2-C
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Abstracts / Atherosclerosis 235 (2014) e84–e191
was calculated as the difference between total HDL-C and HDL3-C. Subjects were recruited in Japan. 670 volunteers who did not have a history of CAD/CHD were invited to participate. Subjects were partitioned according to the following four parameters: male, female, young and old. Results: The Shapiro-Wilk test revealed that all sub-groups were nonparametric. Therefore, the Wilcoxon rank-sum test was used to compare the distribution of variables among sub-groups for HDL3-C, HDL2-C, total HDL-C, and HDL2-C/HDL3-C ratio. Females had significantly higher HDL3-C, HDL2-C, total HDL-C, and HDL2-C/HDL3-C ratios compared to males. Regarding age, HDL2-C and total HDL-C did not show any significant differences, whereas older group showed higher HDL3-C and lower HDL2-C/HDL3-C ratios than younger group. Consequently, only sex difference was found in HDL2-C, whereas both sex and age differences were identified in HDL3-C. Conclusion: Our analysis suggests that HDL subclasses can identify the CHD risk more accurately than total HDL-C. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0256. ASSOCIATION OF INCREASED LARGE HDL PARTICLES WITH IMPROVED CARDIOPULMONARY FITNESS IN ACUTE CORONARY SYNDROME PATIENTS PARTICIPATED WITH EXERCISE-BASED CARDIAC REHABILITATION Y. Itoa, S. Kobab, Y. Yokotab, T. Hiranoc, M. Shojib, Y. Kobayashib a Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan; b The Department of Medicine Division of cardiology, Showa University School of Medicine, Tokyo, Japan; c The Department of Medicine Division of Diabetes Metabolism and Endocrinology, Showa University School of Medicine, Tokyo, Japan
Objectives: Exercised-based cardiac rehabilitation (CR) can increase HDLcholesterol (HDL-C). However it remains unclear how elevated HDL-C and changes of HDL subfractions are correlated with the improvement of exercise tolerance in acute coronary syndrome (ACS) patients participated with CR. Methods: Concentrations of cholesterol and apolipoproteins (Apo) in HDL subfractions separated by heparin-Mn precipitation method were measured at the onset of ACS and at the end of the 6-month CR program in patients (45 men and 6 women) aged of 64.3 11.8 years. All patients received successful percutaneous coronary intervention on admission, and then started to take statins. Results: Serum levels of HDL-C and ApoA1, and concentrations of cholesterol and ApoA1 in large HDL fraction (HDL2) were significantly increased by CR (42.7 mg/dl 14.1 mg/dl to 47.4 mg/dl 14.2 mg/dl, 128.8 mg/dl 23.4 mg/dl to 139.2 mg/dl 26.0 mg/dl, 25.3 mg/dl 12.4 mg/dl to 30.3 mg/dl 13.6 mg/dl, 67.8 mg/dl 20.3 mg/dl to 79.4 mg/dl 24.5 mg/dl respectively), while cholesterol and ApoA1 in small HDL fraction (HDL3) were not changed. Moreover HDL2-C / HDL-C ratio, and HDL2-ApoA1 / ApoA1ratio were significantly increased by CR. In addition, Spearman's rank correlation coefficient analysis revealed that only % increases of HDL2-C were significantly associated with % increased of peak oxygen consumption (VO2) (r ¼ 0.439, p ¼ 0.007), while neither HDL-C nor HDL3C were associated with them. Conclusion: CR can markedly increase the number of HDL2 particles, which is significantly associated with the improvement of cardiopulmonary fitness. These results suggest that CR is very useful therapy for the reverse cholesterol transport, and the secondary prevention. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0637. DEVELOPMENT OF AN AUTOMATED ASSAY FOR APOE-CONTAINING HDL-CHOLESTEROL a
a
b
c
a
a
Y. Takahashi , T. Sakurai , M. Fujikawa , A. Nagasaka , S. Hui , S. Jin , S. Takedaa, H. Fudaa, Y. Itod, H. Chibaa
a
Faculty of Health Sciences, Hokkaido University, Sapporo, Japan; Department of Clinical Laboratory, Sapporo City General Hospital, Sapporo, Japan; c Department of Infectious Diseases, Sapporo City General Hospital, Sapporo, Japan; d Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan b
Objectives: HDL is classified into apoE-containing HDL and apoE-deficient HDL based on the apoE content. Compared to apoE-deficient HDL, apoEcontaining HDL has a stronger activity in cholesterol efflux from macrophage and also in inhibiting platelet aggregation, suggesting potentially anti-atherogenic nature of apoE-containing HDL. Besides, this HDL subclass is known to increase in plasma when CETP inhibitors are administered in animals and human. For quantitative measurement of apoE-containing HDL cholesterol, however, a reported manual assay based on precipitation has been used in spite of poor precision and complicated procedures. We newly developed a 2-step automated assay to quantify apoE-containing HDL cholesterol. This assay is applicable to a wide range of automated analyzers. Methods: We screened surfactants for those with different affinities between apoE-containing HDL and apoE-deficient HDL. As reference methods, precipitation using 13% PEG was used for total HDL-C, precipitation with PT-DS-Mg++ was used for apoE-deficient HDL-C, and apoEcontaining HDL-C was obtained as the difference between apoE-deficient HDL-C and total HDL-C. Results: We found that a polyoxyethylene benzylphenyl ether derivative specifically reacts only with HDL among plasma lipoproteins. Moreover, this surfactant reacted only with apoE-deficient HDL when used at a low concentration. This surfactant is used in the automated assay for apoEcontaining HDL-C. In the 1st step, apoE-deficient HDL is decomposed by a low concentration of the surfactant. And then, cholesterol released from apoE-deficient HDL is degraded to water and oxygen by actions of cholesterol esterase, cholesterol oxidase and catalase. In the 2nd step, apoE-containing HDL is decomposed by a high concentration of the surfactant, and then, hydrogen peroxides derived from apoE-containing HDLC develop a purple-red color. Our automated assay exhibited a good correlation with the reference method (r¼0.84) and good within-run precision (CVs<1.0 %). Conclusion: We developed a fully automated assay to quantify apoE-containing HDL-C. Our new assay can be useful for revealing yet unelucidated clinical utility of apoE-containing HDL-C. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0801. IN BOTH GENDERS, HIGH RATE UNESTERIFIED CHOLESTEROL TRANSFER TO HDL IN THE PLASMA CAN BE PROTECTIVE AGAINST CORONARY ARTERY DISEASE M.C.O. Sprandela, W.A. Huebb, A.C. Segreb, O.G. Laverdya, A.C. Leite-Júniora, F.R. Freitasa, R. Kalil-Filhob, R.C. Maranhãoa a Laboratory of Lipid Metabolism, Heart Institute - Medical School Hospital University of Sao Paulo, São Paulo, Brazil; b Department of Atherosclerosis, Heart Institute - Medical School Hospital - University of Sao Paulo, São Paulo, Brazil
Objectives: New tools to test metabolic and functional aspects of HDL are important for the understanding of HDL anti-atherogenic role and set up new CAD markers. Recently we developed an in vitro assay to assess the transfer of lipids to HDL, a process mediated by transfer proteins (CETP and PLTP) that is crucial in HDL metabolism and reverse cholesterol transport. Here, the assay was used to detect gender differences in lipid transfers in patients with CAD versus non-CAD, since females are less prone to develop CAD than males. Methods: 39 male and 29 female CAD patients and 38 males and 38 females without CAD, all confirmed by cineangiography and age-paired, were enrolled. Fasting plasma samples were incubated with a donor lipidic nanoemulsion labeled with 3H-cholesteryl-esters and 14C-phospholipids or
E. Shakhtshneidera, Y. Raginob, Y. Polonskayab, E. Kashtanovab, Y. Nikitinc, S. Malyutinac, M. Voevodaa a Laboratory of Genetics of Internal Medicine, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia; b Laboratory of Biochemistry, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia; c Laboratory of Internal Medicine, Federal State Budgetary Institution of Internal and Preventive Medicine Siberian, Novosibirsk, Russia
Objectives: We have analyzed the frequencies of HindIII polymorphism of lipoprotein lipase gene (LPL) and lipid profile in Caucasian population of West Siberia and the groups this high and low total cholesterol (TC) level. Methods: The patients included in the analyses were selected based on TC level from population sample surveyed in frame of HAPIEE project (w9000 participants, aged 45-69, men 50%). Totally 259 patients with highest TC level (>300mg/dl) and 228 patients with lowest TC level (<200mg/dl) and 170 randomly selected patients (mean TC level - 235.843.9mg/dl) were included. The differences of TC level between groups are significant. The plasma lipid levels were determined by standard enzymatic assays. The subfractional profile of LDL was determined by method of electrophoresis. The HindIII polymorphism (22125T/G) of LPL gene was analyzed by RELFPCR. Results: Frequencies of H+H+, H+H- and H-H- genotypes were 61%, 35% and 4% in population. The frequency of H- allele was 0.22, 0.28 and 0.29 in HAPIEE population, low and highest TC level groups, respectively (p>0.05). We have found the association of HindIII polymorphism of LPL gene with TG level in HAPIEE population (p¼0.01). We have found the association of polymorphism HindIII of gene LPL with minor peak on a descending curve of subfractional profile LDL (p¼0,02). Conclusion: The Caucasian population of West Siberia is not significantly differs from populations of Europe by frequencies of alleles and genotypes. The genotype H+H+ of HindIII polymorphism of LPL gene has been associated with high TG level. The reported study was partially supported by RFBR, research project N1404-01594a. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0119. HDL AS A BIOMARKER OF REJECTED HEART PRODUCING SEVERE ATHEROSCLEROSIS OF THE TRANSPLANTED HEART W. Gonzáleza, P. Altieria, H.L. Banchsa, D. Iravedraa, I.F. González-Cancelb, R. Calderóna, C. Gurreab, N. Escobalesa, M. Crespoa a Medicine and Physiology, University of Puerto Rico Medical Sciences Campus, San Juan, Puerto Rico; b Heart Transplant Program, Cardiovascular Center of Puerto Rico and the Caribbean, San Juan, Puerto Rico
Objectives: To study the metabolic HDL changes post heart transplant. Methods: 101 Patients (P.) with heart transplant were studied. 65 P. were males and 36 were female, with a mean age of 51 years. Results: Pre and post metabolic changes were: 1. BMI (25 4 – 28 Kg/m2 P < 0.05). 2. Systolic blood pressure (107 13 – 131 20 P <0.05). 3. Diastolic blood pressure (70 13 – 81 10 mmHg). 4. Fasting blood sugar (107 37 – 117 55 mg%) N.S. 5. Total cholesterol (170 55 – 189 32mg/dl) P < 0.05. 6. Total HDL (38 16 – 52 17mg/dl) P < 0.05. 7. Total LDL (99 20 – 83 15mg/dl) N.S. 8. Triglycerides (163 10 – 188 12mg/dl) N.S. Conclusion: Our data shows that all the P. developed Mets. The HDL of the whole groups increased from 38 mg/dl to 52 mg/dl. These changes are not explained only by the immunosuppressive treatment, HDL has a role as an anti-inflammatory agent which is inherent in the lipoprotein characteristics. 16 P. died of rejection. The HDL of the rejected P. increased from 47 22 to 71 40 mg/dl P. < 0.007. This increase persisted through the course
e101
of the rejection period. The mean age of the transplanted heart was 22 years. The rejection occurred in less than 5 years. (3.5 years). Seven autopsies were done. Five of them showed severe atherosclerotic changes in the coronaries and aorta. This is probably explained on basis of dysfunctional HDL. Probably the high levels and persistent elevation of HDL in the rejected group can be used as a biomarker to predict rejection. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0807. HDL, ITS SUBFRACTIONS AND APOLIPOPROTEIN-A1 AS PREDICTORS OF HIGH PLATELET REACTIVITY AND MAJOR ADVERSE CARDIOVASCULAR EVENTS IN PATIENTS WITH STEMI R. Grzybczak, A. Siniarski, J. Zalewski, J. Nessler, G. Gajos Department of Coronary Disease John Paul II Hospital, Jagiellonian University Medical College, Krakow, Poland Objectives: Determine the relationship between serum HDL-cholesterol levels and its subfractions (LpA-I and LpA-I/A-II) and apoprotein A1 (Apo A-I) and early high platelet reactivity (HPR) on clopidogrel in the acute phase of STEMI. Methods: Fifty-two patients (14 women and 38 men) aged 60.6 9.14 years old with STEMI were included in this study. Patients (pts) were treated with primary PCI with stent implantation and optimal medical therapy including aspirin and clopidogrel for 12 months. After 12 hours of the STEMI onset we assessed: HDL and its subfractions (enzymatic and rocket immunoelectrophoresis, respectively); platelet reactivity (optical aggregometry and flow cytofluorometry). The study group was divided into 2 subgroups: with high and low levels of HDL-C, Apo A-I, LpA-I and LpA-I/A-II according to post-hoc analysis of the median concentration. During 12-months followup we assessed the incidence of major adverse cardiovascular events. Results: Higher values of LpA-I and lower concentrations of LpA-I/A-II were associated with HPR despite clopidogrel treatment. LpA-I levels higher than 57.52 mg/dl predicted HPR. The risk of inadequate response to antiplatelet therapy increased by 3% with every 1 mg/dl increase of LpA-I level. Moreover, HDL-C and Apo A-I over 1.19 mmol/l and 129.6 mg/dl respectively were significant predictors of rehospitalization in 12-month followup. The cut-off point 4 mg/dl for D12 LpA-I/A-II by the ROC curve analysis most accurately predicted rehospitalization with 66% specificity and 100% sensitivity. Moreover, LpA-I over 58.27 mg/dl was associated with higher risk of unplanned coronary revascularization. Conclusion: Increased LpA-I concentration predicted HPR despite antiplatelet treatment in pts with acute STEMI. Higher LpA-I levels indicated also increased risk of major cardiovascular events. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0631. ESTABLISHMENT OF REFERENCE RANGE FOR HDL SUBFRACTIONS IN JAPANESE POPULATION WITH A NEW AUTOMATED ASSAY Y. Ito, N. Satoh, T. Ishii, J. Kumakura Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan Objectives: Measurement of high-density lipoprotein (HDL) 2 and HDL3 subfractions might be more useful for evaluating coronary risk than total HDL-cholesterol (C). However, methods of measuring HDL2 and HDL3 are quite laborious for general clinical use and thus so far it has been difficult to establish reference ranges. We carried out a study to establish a reference range for HDL subfractions in Japanese population using our novel homogeneous assay for HDL subfractions. Methods: HDL-C and HDL3-C were measured by our homogeneous assays on a Hitachi 917 automated clinical chemistry analyzer (Hitachi). HDL2-C
e102
Abstracts / Atherosclerosis 235 (2014) e84–e191
was calculated as the difference between total HDL-C and HDL3-C. Subjects were recruited in Japan. 670 volunteers who did not have a history of CAD/CHD were invited to participate. Subjects were partitioned according to the following four parameters: male, female, young and old. Results: The Shapiro-Wilk test revealed that all sub-groups were nonparametric. Therefore, the Wilcoxon rank-sum test was used to compare the distribution of variables among sub-groups for HDL3-C, HDL2-C, total HDL-C, and HDL2-C/HDL3-C ratio. Females had significantly higher HDL3-C, HDL2-C, total HDL-C, and HDL2-C/HDL3-C ratios compared to males. Regarding age, HDL2-C and total HDL-C did not show any significant differences, whereas older group showed higher HDL3-C and lower HDL2-C/HDL3-C ratios than younger group. Consequently, only sex difference was found in HDL2-C, whereas both sex and age differences were identified in HDL3-C. Conclusion: Our analysis suggests that HDL subclasses can identify the CHD risk more accurately than total HDL-C. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0256. ASSOCIATION OF INCREASED LARGE HDL PARTICLES WITH IMPROVED CARDIOPULMONARY FITNESS IN ACUTE CORONARY SYNDROME PATIENTS PARTICIPATED WITH EXERCISE-BASED CARDIAC REHABILITATION Y. Itoa, S. Kobab, Y. Yokotab, T. Hiranoc, M. Shojib, Y. Kobayashib a Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan; b The Department of Medicine Division of cardiology, Showa University School of Medicine, Tokyo, Japan; c The Department of Medicine Division of Diabetes Metabolism and Endocrinology, Showa University School of Medicine, Tokyo, Japan
Objectives: Exercised-based cardiac rehabilitation (CR) can increase HDLcholesterol (HDL-C). However it remains unclear how elevated HDL-C and changes of HDL subfractions are correlated with the improvement of exercise tolerance in acute coronary syndrome (ACS) patients participated with CR. Methods: Concentrations of cholesterol and apolipoproteins (Apo) in HDL subfractions separated by heparin-Mn precipitation method were measured at the onset of ACS and at the end of the 6-month CR program in patients (45 men and 6 women) aged of 64.3 11.8 years. All patients received successful percutaneous coronary intervention on admission, and then started to take statins. Results: Serum levels of HDL-C and ApoA1, and concentrations of cholesterol and ApoA1 in large HDL fraction (HDL2) were significantly increased by CR (42.7 mg/dl 14.1 mg/dl to 47.4 mg/dl 14.2 mg/dl, 128.8 mg/dl 23.4 mg/dl to 139.2 mg/dl 26.0 mg/dl, 25.3 mg/dl 12.4 mg/dl to 30.3 mg/dl 13.6 mg/dl, 67.8 mg/dl 20.3 mg/dl to 79.4 mg/dl 24.5 mg/dl respectively), while cholesterol and ApoA1 in small HDL fraction (HDL3) were not changed. Moreover HDL2-C / HDL-C ratio, and HDL2-ApoA1 / ApoA1ratio were significantly increased by CR. In addition, Spearman's rank correlation coefficient analysis revealed that only % increases of HDL2-C were significantly associated with % increased of peak oxygen consumption (VO2) (r ¼ 0.439, p ¼ 0.007), while neither HDL-C nor HDL3C were associated with them. Conclusion: CR can markedly increase the number of HDL2 particles, which is significantly associated with the improvement of cardiopulmonary fitness. These results suggest that CR is very useful therapy for the reverse cholesterol transport, and the secondary prevention. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0637. DEVELOPMENT OF AN AUTOMATED ASSAY FOR APOE-CONTAINING HDL-CHOLESTEROL a
a
b
c
a
a
Y. Takahashi , T. Sakurai , M. Fujikawa , A. Nagasaka , S. Hui , S. Jin , S. Takedaa, H. Fudaa, Y. Itod, H. Chibaa
a
Faculty of Health Sciences, Hokkaido University, Sapporo, Japan; Department of Clinical Laboratory, Sapporo City General Hospital, Sapporo, Japan; c Department of Infectious Diseases, Sapporo City General Hospital, Sapporo, Japan; d Research and Development Department, Denka Seiken Co. Ltd., Tokyo, Japan b
Objectives: HDL is classified into apoE-containing HDL and apoE-deficient HDL based on the apoE content. Compared to apoE-deficient HDL, apoEcontaining HDL has a stronger activity in cholesterol efflux from macrophage and also in inhibiting platelet aggregation, suggesting potentially anti-atherogenic nature of apoE-containing HDL. Besides, this HDL subclass is known to increase in plasma when CETP inhibitors are administered in animals and human. For quantitative measurement of apoE-containing HDL cholesterol, however, a reported manual assay based on precipitation has been used in spite of poor precision and complicated procedures. We newly developed a 2-step automated assay to quantify apoE-containing HDL cholesterol. This assay is applicable to a wide range of automated analyzers. Methods: We screened surfactants for those with different affinities between apoE-containing HDL and apoE-deficient HDL. As reference methods, precipitation using 13% PEG was used for total HDL-C, precipitation with PT-DS-Mg++ was used for apoE-deficient HDL-C, and apoEcontaining HDL-C was obtained as the difference between apoE-deficient HDL-C and total HDL-C. Results: We found that a polyoxyethylene benzylphenyl ether derivative specifically reacts only with HDL among plasma lipoproteins. Moreover, this surfactant reacted only with apoE-deficient HDL when used at a low concentration. This surfactant is used in the automated assay for apoEcontaining HDL-C. In the 1st step, apoE-deficient HDL is decomposed by a low concentration of the surfactant. And then, cholesterol released from apoE-deficient HDL is degraded to water and oxygen by actions of cholesterol esterase, cholesterol oxidase and catalase. In the 2nd step, apoE-containing HDL is decomposed by a high concentration of the surfactant, and then, hydrogen peroxides derived from apoE-containing HDLC develop a purple-red color. Our automated assay exhibited a good correlation with the reference method (r¼0.84) and good within-run precision (CVs<1.0 %). Conclusion: We developed a fully automated assay to quantify apoE-containing HDL-C. Our new assay can be useful for revealing yet unelucidated clinical utility of apoE-containing HDL-C. 06 - New protective effects of HDL on the cardiovascular system and beyond EAS-0801. IN BOTH GENDERS, HIGH RATE UNESTERIFIED CHOLESTEROL TRANSFER TO HDL IN THE PLASMA CAN BE PROTECTIVE AGAINST CORONARY ARTERY DISEASE M.C.O. Sprandela, W.A. Huebb, A.C. Segreb, O.G. Laverdya, A.C. Leite-Júniora, F.R. Freitasa, R. Kalil-Filhob, R.C. Maranhãoa a Laboratory of Lipid Metabolism, Heart Institute - Medical School Hospital University of Sao Paulo, São Paulo, Brazil; b Department of Atherosclerosis, Heart Institute - Medical School Hospital - University of Sao Paulo, São Paulo, Brazil
Objectives: New tools to test metabolic and functional aspects of HDL are important for the understanding of HDL anti-atherogenic role and set up new CAD markers. Recently we developed an in vitro assay to assess the transfer of lipids to HDL, a process mediated by transfer proteins (CETP and PLTP) that is crucial in HDL metabolism and reverse cholesterol transport. Here, the assay was used to detect gender differences in lipid transfers in patients with CAD versus non-CAD, since females are less prone to develop CAD than males. Methods: 39 male and 29 female CAD patients and 38 males and 38 females without CAD, all confirmed by cineangiography and age-paired, were enrolled. Fasting plasma samples were incubated with a donor lipidic nanoemulsion labeled with 3H-cholesteryl-esters and 14C-phospholipids or