title: A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
abstract: Current cell-based assays for determining the functional properties of high-density lipoproteins (HDL) have limitations. We report here the development of a new, robust fluorometric cell-free biochemical assay that measures HDL lipid peroxidation (HDLox) based on the oxidation of the fluorochrome Amplex Red. HDLox correlated with previously validated cell-based (r = 0.47, p<0.001) and cell-free assays (r = 0.46, p<0.001). HDLox distinguished dysfunctional HDL in established animal models of atherosclerosis and Human Immunodeficiency Virus (HIV) patients. Using an immunoaffinity method for capturing HDL, we demonstrate the utility of this novel assay for measuring HDLox in a high throughput format. Furthermore, HDLox correlated significantly with measures of cardiovascular diseases including carotid intima media thickness (r = 0.35, p<0.01) and subendocardial viability ratio (r = -0.21, p = 0.05) and physiological parameters such as metabolic and anthropometric parameters (p<0.05). In conclusion, we report the development of a new fluorometric method that offers a reproducible and rapid means for determining HDL function/quality that is suitable for high throughput implementation.
authors.affiliation: Department of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Theodoros
authors.initials: T
authors.lastname: Kelesidis
authors.affiliation: Exercise and Metabolic Disease Research Laboratory, Translational Sciences Section, School of Nursing, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Christian K
authors.initials: CK
authors.lastname: Roberts
authors.affiliation: Department of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Diana
authors.initials: D
authors.lastname: Huynh
authors.affiliation: Department of Obstetrics and Gynecology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America; Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Otoniel
authors.initials: O
authors.lastname: Martínez-Maza
authors.affiliation: Department of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Judith S
authors.initials: JS
authors.lastname: Currier
authors.affiliation: Department of Medicine, University of California Los Angeles, Los Angeles, California, United States of America; Department of Obstetrics and Gynecology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America; Department of Molecular and Medical Pharmacology, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Srinivasa T
authors.initials: ST
authors.lastname: Reddy
authors.affiliation: Department of Medicine, University of California Los Angeles, Los Angeles, California, United States of America; Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Otto O
authors.initials: OO
authors.lastname: Yang
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D059168:Carotid Intima-Media Thickness; D003324:Coronary Artery Disease / Q000097:blood / Q000378:metabolism; D005260:Female; D005470:Fluorometry / Q000379:methods*; D015658:HIV Infections / Q000097:blood / Q000378:metabolism; D015497:HIV-1 / Q000302:isolation & purification; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D015227:Lipid Peroxidation; D008075:Lipoproteins, HDL / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D010078:Oxazines / Q000032:analysis / Q000378:metabolism; D055815:Young Adult
chemical_list: D008075:Lipoproteins, HDL; D010078:Oxazines; C470430:Amplex Red