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We retrospectively analyzed data on patients with type 2 diabetes who underwent CCTA and measurements of MDA-LDL at the University of Tsukuba Hospital from April 2009 to March 2012.
Serum MDA-LDL levels were measured by a sandwich enzyme-linked immunosorbent assay (Sekisui Medical, Tokyo, Japan).
MDA-LDL (odds ratio (OR) 1.02 (95% confidence interval 1.00-1.04), P = 0.039), MDA-LDL/LDL-C (1.13 (1.03-1.25), P = 0.013), MDA-LDL/ HDL-C (1.02 (1.00-1.05), P = 0.047), and (MDA-LDL/ LDL)/HDL-C (1.16 (1.03-1.30), P = 0.013) were independent predictors of CAS after adjustments for age, sex, body mass index, hypertension, duration of diabetes, smoking, and HbA1c.
 Nonstandard abbreviafions: oxLDL, oxidized LDL; gLDL, glycated LDL; acLDL, acetylated LDL; ApoB, apolipoprotein B; cLDL, carbamylated LDL; nLDL, native LDL; HRP, horseradish peroxidase; BSA, bovine serum albumin; PBS, phosphate-buttered saline; PBS-E, PBS with EDTA; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; TC, total cholesterol; and MDA-LDL, malondialdehyde-modified LDL.
At the end of the HAT selection period, supernatants of each well were tested for antibodies binding to [Cu.sup.++]-oxidized LDL and/or MDALDL with commercially available ELISA methods (oLAb-ELISA, Biomedica, Vienna, Austria; MDA-LDL IgG/IgM, LDN Ltd., Nordhorn, Germany) adapted for the purposes of this investigation .
MDA-LDL and [Cu.sup.++]-oxidized LDL-coated ELISA plates were allowed to react with either clone 3G5 or subclone 3G5/D4 IgM antibodies.
Competition experiments with other adducts of LDL than MDA or [Cu.sup.++]-oxidized LDL like hexanal-LDL, 4-HNE-LDL, or HOCl-LDL did not show these substances to interfere with the antibody-antigen reaction of the MDA-LDL plate.
Detection of autoantibodies against MDA-LDL remains the most important argument in favor of the oxidative modification of LDL in vivo .
LDL of healthy individuals 2.4 Glycosylated LDL 2.6 Acetylated LDL 2.8 [Cu.sup.2+]-oxidized LDL 3.5 Patients' LDL 11.3 MDA-LDL 10.9 Desialylated LDL 89.4 Adapted from .
Monoclonal antibodies against MDA-LDL (ML25) and apolipoprotein B (apo B;monoclonal antibody AB16) were obtained from Daiichi Pure Chemicals Co.
Duplicate 100-[micro]L portions of the diluted sample were then added to the wells of plates that were coated with monoclonal antibody against MDA-LDL (ML25; 0.8 [micro]g/well).
Primary standard was used with preparative MDA-LDL, in which 15% of the total amino groups were modified.
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