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Influence of HDL-cholesterol levels on HDL particles lipid composition in healthy individuals. an NMR-based lipidomic study
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Abstracts / Atherosclerosis 252 (2016) e1ee196
Objectives: The functionality of high-density lipoprotein (HDL) has been found to be impaired in chronic ischaemic heart failure (HF). However, the relationship between HDL functionality and outcomes in acute HF (AHF) has not been studied. The present study investigates whether the metrics of HDL functionality, including HDL cholesterol efflux capacity and HDLassociated paraoxonase (PON)-1 arylesterase (AE) activity are associated with hospital mortality in AHF patients. Methods: The study was performed as a prospective, single-centre, observational research on 152 patients, defined and categorised according to the ESC and ACCF/AHA Guidelines for HF by time of onset, final clinical presentation and ejection fraction. HDL cholesterol efflux capacity was examined by measuring the capacity of apoB depleted serum to remove tritium-labelled cholesterol from cultured macrophages. The AE activity of the HDL fraction was examined by a photometric assay. Results: The mean age of the included patients (52% female) was 75.2 years (SD 10.3) and hospital mortality was 14.5%. In a univariable regression analysis, low cholesterol efflux, but not AE activity, was significantly associated with hospital mortality [odds ratio (OR) 0.78, 95% confidence interval (CI) 0.64-0.96, p¼0.019]. However, in multivariable analysis progressively adjusting for age, gender, body mass index, LDL, HDL, log(triglycerides) and mean arterial blood pressure, the association obtained for cholesterol efflux capacity and hospital mortality by univariable analysis, despite a stable OR, did not stay significant (OR 0.83, 95% CI 0.65-1.05, p¼0.131). Conclusions: HDL cholesterol efflux capacity, but not AE activity, is associated with hospital mortality in AHF patients. EAS16-0231, LIPOPROTEINS AND LIPID METABOLISM: HDL. CAUSAL LINK BETWEEN SERUM AMYLOID A AND OXIDATIVE DAMAGES TO HIGH-DENSITY LIPOPROTEIN S. Jayaraman 1, C. Haupt 2, O. Gursky 3. 1 Boston University School of Medicine, Physiology & Biophysics, Boston, USA; 2 University of Ulm, Institute for Pharmaceutical Biotechnology, Ulm, Germany; 3 Boston University School of Medicine, Phsyiology & Biophysics, Boston, USA Objectives: High-density lipoproteins (HDL) exert potent anti-atherogenic activities, including anti-oxidative action, which help attenuate the progression of atherosclerosis. During acute and chronic inflammation, such activities can be compromised due to altered protein and lipid composition in HDL. In inflammation, serum amyloid A (SAA) binds HDL and importantly modulates its anti-oxidative and anti-inflammatory properties. Our objective is to unravel the molecular basis for oxidative damage to SAAcontaining HDL. Methods: SAA-enriched HDL (SAA-HDL) were obtained by incubating murine SAA1.1 with human plasma HDL at 37oC, and purified by gel filtration. HDL were oxidized by using chemical (Cu2+, NaOCl) or enzymatic (MPO/H2O2/NaCl) agents. Particle composition and integrity were analyzed by mass spectrometry, SDS and native PAGE, and immunoblotting. Results: No significant structural remodeling of HDL upon oxidation was observed by native PAGE. However, protein analysis showed significant changes. As expected, both normal HDL and SAA-HDL exhibited high-molecular-weight cross-linked proteins, including apoA-I and apoA-II, whose amount progressively increased with increasing the oxidation degree. Surprisingly, SAA-HDL exhibited a unique 40 kDa band that corresponded to cross-linked apoA-IeSAA hetero-dimer. This hetero-dimer appeared in early stages of oxidation, before any significant cross-linking between apoAI and apoA-II was detected. Preliminary studies indicated that this heterodimer formation occurs in HDL-bound but not in lipid-free proteins. Conclusions: We report a novel apoA-IeSAA hetero-dimer formed upon moderate HDL oxidation. A similar hetero-dimer is likely to form in vivo, which is expected to influence functional properties of apoA-I and HDL and potentially contribute to the pro-atherogenic effects of HDL during inflammation. EAS16-0440, LIPOPROTEINS AND LIPID METABOLISM: HDL. INFLUENCE OF HDL-CHOLESTEROL LEVELS ON HDL PARTICLES LIPID COMPOSITION IN HEALTHY INDIVIDUALS. AN NMR-BASED LIPIDOMIC STUDY
C. Kostara 1, V. Tsimihodimos 2, E. Bairaktari 1, M. Elisaf 2. 1 University of Ioannina, Laboratory of Clinical Chemistry- Medical School, Ioannina, Greece; 2 University of Ioannina, Department of Internal Medicine- Medical School, Ioannina, Greece Objectives: Current studies proposed that the protective properties of HDL lipoproteins against cardiovascular events may depend not only on the quantity of cholesterol but on overall particle composition, structure and quality. The investigation of 1H NMR-based lipid profiling of HDL lipoproteins in healthy individuals with low and high serum HDL-cholesterol levels. Methods: Serum samples from 15 healthy individuals with low (<40 mg/ dl), 21 with normal (40-59 mg/dl) and 15 with high HDL-cholesterol levels (>60 mg/dl) were collected after an overnight fast. All groups were matched in the other serum lipid parameters (total and LDL-cholesterol and triglycerides). The lipid content of HDL lipoproteins was extracted and pattern recognition analysis was applied on the 1H NMR lipidomic data. Results: Individuals with low HDL-cholesterol presented with lower phospholipid content (phosphatidylcholine and sphingomyelin), cholesterol, omega-3 and unsaturated fatty acids and higher saturated fatty acid content compared with individuals with normal HDL-cholesterol. On the other hand, HDL particles isolated from those with high HDL-cholesterol were enriched in phospholipids (phosphatidylcholine, sphingomyelin), cholesterol, linoleic acid, omega-3 and unsaturated fatty acids and depleted in triglycerides and saturated fatty acids compared to those from individuals with normal HDL-cholesterol. Conclusions: Individuals with low HDL-cholesterol presented with a more atherogenic lipid profiling in HDL particles, whereas these with high HDLcholesterol presented more atheroprotective lipid profiling compared to those with normal HDL-cholesterol. These changes in composition and structure probably affect the functionality and metabolism of HDL particles and could constitute novel specific biomarkers for the atheroprotective properties of HDL. EAS16-0492, LIPOPROTEINS AND LIPID METABOLISM: HDL. HIGH-DENSITY LIPOPROTEIN (HDL) FUNCTION AND SUBCLASS DISTRIBUTION IN OBESE AND NORMAL-WEIGHT BLACK AND WHITE AFRICAN WOMEN. N. Woudberg 1, J. Goedecke 2, D. Blackhurst 3, M. Frias 4, R. James 4, L. Opie 1, S. Lecour 1. 1 University of Cape Town, Medicine, Cape Town, South Africa; 2 University of Cape Town, Human Biology, Cape Town, South Africa; 3 University of Cape Town, Pathology, Cape Town, South Africa; 4 University of Geneva, Internal Medicine, Geneva, Switzerland Objectives: Obesity and low high-density lipoprotein (HDL) concentrations are associated with cardiovascular risk. Surprisingly, despite a greater prevalence of obesity and lower HDL concentrations than white women, black South African women are relatively protected against certain cardiovascular diseases. However, it is unclear whether this is related to altered HDL function and subclass distribution. Our aim was to therefore investigate HDL function and subclass in a sample of 40 black and white, normal-weight and obese South African women Methods: HDL functionality was assessed by means of paraoxonase (PON1) activity, platelet activating factor acetylhydrolase (PAF-AH) activity, Oxygen Radical Absorbance Capacity (ORAC) and quantification of the expression of vascular cell adhesion molecule in endothelial cells. PON-1 and PAF-AH expression was determined in isolated HDL using Western blotting. Levels of large, intermediate and small HDL subclasses were measured using the Lipoprint® system. Results: PON-1 activity was lower in white compared to black women, regardless of PON-1 protein levels (0.49±0.09 U/L vs 0.78±0.10 U/L, p<0.05), while obese black women had lower PAF-AH activity and HDLassociated PAF-AH expression compared to white obese women (9.34±1.15 U/L vs 13.89±1.21 U/L, p <0.05). Compared to normal-weight women, obese women had lower large HDL, greater intermediate and small HDL; however subclasses did not differ by ethnicity. Conclusions: Our data show that both obesity and ethnicity are associated with differences in HDL functionality, and obesity was associated with decreases in large HDL subclass distribution. HDL functionality and
Abstracts / Atherosclerosis 252 (2016) e1ee196
Objectives: The functionality of high-density lipoprotein (HDL) has been found to be impaired in chronic ischaemic heart failure (HF). However, the relationship between HDL functionality and outcomes in acute HF (AHF) has not been studied. The present study investigates whether the metrics of HDL functionality, including HDL cholesterol efflux capacity and HDLassociated paraoxonase (PON)-1 arylesterase (AE) activity are associated with hospital mortality in AHF patients. Methods: The study was performed as a prospective, single-centre, observational research on 152 patients, defined and categorised according to the ESC and ACCF/AHA Guidelines for HF by time of onset, final clinical presentation and ejection fraction. HDL cholesterol efflux capacity was examined by measuring the capacity of apoB depleted serum to remove tritium-labelled cholesterol from cultured macrophages. The AE activity of the HDL fraction was examined by a photometric assay. Results: The mean age of the included patients (52% female) was 75.2 years (SD 10.3) and hospital mortality was 14.5%. In a univariable regression analysis, low cholesterol efflux, but not AE activity, was significantly associated with hospital mortality [odds ratio (OR) 0.78, 95% confidence interval (CI) 0.64-0.96, p¼0.019]. However, in multivariable analysis progressively adjusting for age, gender, body mass index, LDL, HDL, log(triglycerides) and mean arterial blood pressure, the association obtained for cholesterol efflux capacity and hospital mortality by univariable analysis, despite a stable OR, did not stay significant (OR 0.83, 95% CI 0.65-1.05, p¼0.131). Conclusions: HDL cholesterol efflux capacity, but not AE activity, is associated with hospital mortality in AHF patients. EAS16-0231, LIPOPROTEINS AND LIPID METABOLISM: HDL. CAUSAL LINK BETWEEN SERUM AMYLOID A AND OXIDATIVE DAMAGES TO HIGH-DENSITY LIPOPROTEIN S. Jayaraman 1, C. Haupt 2, O. Gursky 3. 1 Boston University School of Medicine, Physiology & Biophysics, Boston, USA; 2 University of Ulm, Institute for Pharmaceutical Biotechnology, Ulm, Germany; 3 Boston University School of Medicine, Phsyiology & Biophysics, Boston, USA Objectives: High-density lipoproteins (HDL) exert potent anti-atherogenic activities, including anti-oxidative action, which help attenuate the progression of atherosclerosis. During acute and chronic inflammation, such activities can be compromised due to altered protein and lipid composition in HDL. In inflammation, serum amyloid A (SAA) binds HDL and importantly modulates its anti-oxidative and anti-inflammatory properties. Our objective is to unravel the molecular basis for oxidative damage to SAAcontaining HDL. Methods: SAA-enriched HDL (SAA-HDL) were obtained by incubating murine SAA1.1 with human plasma HDL at 37oC, and purified by gel filtration. HDL were oxidized by using chemical (Cu2+, NaOCl) or enzymatic (MPO/H2O2/NaCl) agents. Particle composition and integrity were analyzed by mass spectrometry, SDS and native PAGE, and immunoblotting. Results: No significant structural remodeling of HDL upon oxidation was observed by native PAGE. However, protein analysis showed significant changes. As expected, both normal HDL and SAA-HDL exhibited high-molecular-weight cross-linked proteins, including apoA-I and apoA-II, whose amount progressively increased with increasing the oxidation degree. Surprisingly, SAA-HDL exhibited a unique 40 kDa band that corresponded to cross-linked apoA-IeSAA hetero-dimer. This hetero-dimer appeared in early stages of oxidation, before any significant cross-linking between apoAI and apoA-II was detected. Preliminary studies indicated that this heterodimer formation occurs in HDL-bound but not in lipid-free proteins. Conclusions: We report a novel apoA-IeSAA hetero-dimer formed upon moderate HDL oxidation. A similar hetero-dimer is likely to form in vivo, which is expected to influence functional properties of apoA-I and HDL and potentially contribute to the pro-atherogenic effects of HDL during inflammation. EAS16-0440, LIPOPROTEINS AND LIPID METABOLISM: HDL. INFLUENCE OF HDL-CHOLESTEROL LEVELS ON HDL PARTICLES LIPID COMPOSITION IN HEALTHY INDIVIDUALS. AN NMR-BASED LIPIDOMIC STUDY
C. Kostara 1, V. Tsimihodimos 2, E. Bairaktari 1, M. Elisaf 2. 1 University of Ioannina, Laboratory of Clinical Chemistry- Medical School, Ioannina, Greece; 2 University of Ioannina, Department of Internal Medicine- Medical School, Ioannina, Greece Objectives: Current studies proposed that the protective properties of HDL lipoproteins against cardiovascular events may depend not only on the quantity of cholesterol but on overall particle composition, structure and quality. The investigation of 1H NMR-based lipid profiling of HDL lipoproteins in healthy individuals with low and high serum HDL-cholesterol levels. Methods: Serum samples from 15 healthy individuals with low (<40 mg/ dl), 21 with normal (40-59 mg/dl) and 15 with high HDL-cholesterol levels (>60 mg/dl) were collected after an overnight fast. All groups were matched in the other serum lipid parameters (total and LDL-cholesterol and triglycerides). The lipid content of HDL lipoproteins was extracted and pattern recognition analysis was applied on the 1H NMR lipidomic data. Results: Individuals with low HDL-cholesterol presented with lower phospholipid content (phosphatidylcholine and sphingomyelin), cholesterol, omega-3 and unsaturated fatty acids and higher saturated fatty acid content compared with individuals with normal HDL-cholesterol. On the other hand, HDL particles isolated from those with high HDL-cholesterol were enriched in phospholipids (phosphatidylcholine, sphingomyelin), cholesterol, linoleic acid, omega-3 and unsaturated fatty acids and depleted in triglycerides and saturated fatty acids compared to those from individuals with normal HDL-cholesterol. Conclusions: Individuals with low HDL-cholesterol presented with a more atherogenic lipid profiling in HDL particles, whereas these with high HDLcholesterol presented more atheroprotective lipid profiling compared to those with normal HDL-cholesterol. These changes in composition and structure probably affect the functionality and metabolism of HDL particles and could constitute novel specific biomarkers for the atheroprotective properties of HDL. EAS16-0492, LIPOPROTEINS AND LIPID METABOLISM: HDL. HIGH-DENSITY LIPOPROTEIN (HDL) FUNCTION AND SUBCLASS DISTRIBUTION IN OBESE AND NORMAL-WEIGHT BLACK AND WHITE AFRICAN WOMEN. N. Woudberg 1, J. Goedecke 2, D. Blackhurst 3, M. Frias 4, R. James 4, L. Opie 1, S. Lecour 1. 1 University of Cape Town, Medicine, Cape Town, South Africa; 2 University of Cape Town, Human Biology, Cape Town, South Africa; 3 University of Cape Town, Pathology, Cape Town, South Africa; 4 University of Geneva, Internal Medicine, Geneva, Switzerland Objectives: Obesity and low high-density lipoprotein (HDL) concentrations are associated with cardiovascular risk. Surprisingly, despite a greater prevalence of obesity and lower HDL concentrations than white women, black South African women are relatively protected against certain cardiovascular diseases. However, it is unclear whether this is related to altered HDL function and subclass distribution. Our aim was to therefore investigate HDL function and subclass in a sample of 40 black and white, normal-weight and obese South African women Methods: HDL functionality was assessed by means of paraoxonase (PON1) activity, platelet activating factor acetylhydrolase (PAF-AH) activity, Oxygen Radical Absorbance Capacity (ORAC) and quantification of the expression of vascular cell adhesion molecule in endothelial cells. PON-1 and PAF-AH expression was determined in isolated HDL using Western blotting. Levels of large, intermediate and small HDL subclasses were measured using the Lipoprint® system. Results: PON-1 activity was lower in white compared to black women, regardless of PON-1 protein levels (0.49±0.09 U/L vs 0.78±0.10 U/L, p<0.05), while obese black women had lower PAF-AH activity and HDLassociated PAF-AH expression compared to white obese women (9.34±1.15 U/L vs 13.89±1.21 U/L, p <0.05). Compared to normal-weight women, obese women had lower large HDL, greater intermediate and small HDL; however subclasses did not differ by ethnicity. Conclusions: Our data show that both obesity and ethnicity are associated with differences in HDL functionality, and obesity was associated with decreases in large HDL subclass distribution. HDL functionality and