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                <title xml:lang="en">Small, dense high-density lipoprotein-3 particles are enriched in negatively charged phospholipids: relevance to cellular cholesterol efflux, antioxidative, antithrombotic, anti-inflammatory, and antiapoptotic functionalities.</title>
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                <title level="j">Arteriosclerosis, Thrombosis, and Vascular Biology</title>
                <imprint>
                  <publisher>American Heart Association</publisher>
                  <biblScope unit="volume">33</biblScope>
                  <biblScope unit="issue">12</biblScope>
                  <biblScope unit="pp">2715-23</biblScope>
                  <date type="datePub">2013-12</date>
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              <classCode scheme="mesh">Inflammation</classCode>
              <classCode scheme="mesh">Inflammation Mediators</classCode>
              <classCode scheme="mesh">Lipoproteins, HDL3</classCode>
              <classCode scheme="mesh">Lipoproteins, LDL</classCode>
              <classCode scheme="mesh">Macrophages</classCode>
              <classCode scheme="mesh">Male</classCode>
              <classCode scheme="mesh">Middle Aged</classCode>
              <classCode scheme="mesh">Oxidation-Reduction</classCode>
              <classCode scheme="mesh">Oxidative Stress</classCode>
              <classCode scheme="mesh">Apoptosis</classCode>
              <classCode scheme="mesh">Particle Size</classCode>
              <classCode scheme="mesh">Phospholipids</classCode>
              <classCode scheme="mesh">Sphingolipids</classCode>
              <classCode scheme="mesh">Tandem Mass Spectrometry</classCode>
              <classCode scheme="mesh">Thrombosis</classCode>
              <classCode scheme="mesh">Adult</classCode>
              <classCode scheme="mesh">Atherosclerosis</classCode>
              <classCode scheme="mesh">Blood Platelets</classCode>
              <classCode scheme="mesh">Cell Line, Tumor</classCode>
              <classCode scheme="mesh">Ceramides</classCode>
              <classCode scheme="mesh">Cholesterol</classCode>
              <classCode scheme="mesh">Chromatography, Liquid</classCode>
              <classCode scheme="mesh">Endothelial Cells</classCode>
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              <p>OBJECTIVE: High-density lipoprotein (HDL) displays multiple atheroprotective activities and is highly heterogeneous in structure, composition, and function; the molecular determinants of atheroprotective functions of HDL are incompletely understood. Because phospholipids represent a major bioactive lipid component of HDL, we characterized the phosphosphingolipidome of major normolipidemic HDL subpopulations and related it to HDL functionality. APPROACH AND RESULTS: Using an original liquid chromatography-mass spectrometry/mass spectrometry methodology for phospholipid and sphingolipid profiling, 162 individual molecular lipid species were quantified across the 9 lipid subclasses, in the order of decreasing abundance, phosphatidylcholine&gt;sphingomyelin&gt;lysophosphatidylcholine&gt;phosphatidylethanolamine&gt;phosphatidylinositol&gt;ceramide&gt;phosphatidylserine&gt;phosphatidylglycerol&gt;phosphatidic acid. When data were expressed relative to total lipid, the contents of lysophosphatidylcholine and of negatively charged phosphatidylserine and phosphatidic acid increased progressively with increase in hydrated density of HDL, whereas the proportions of sphingomyelin and ceramide decreased. Key biological activities of HDL subpopulations, notably cholesterol efflux capacity from human THP-1 macrophages, antioxidative activity toward low-density lipoprotein oxidation, antithrombotic activity in human platelets, cell-free anti-inflammatory activity, and antiapoptotic activity in endothelial cells, were predominantly associated with small, dense, protein-rich HDL3. The biological activities of HDL particles were strongly intercorrelated, exhibiting significant correlations with multiple components of the HDL phosphosphingolipidome. Specifically, the content of phosphatidylserine revealed positive correlations with all metrics of HDL functionality, reflecting enrichment of phosphatidylserine in small, dense HDL3. CONCLUSIONS: Our structure-function analysis thereby reveals that the HDL lipidome may strongly affect atheroprotective functionality.</p>
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