Cholesterol efflux capacity.

<p>Apolipoprotein B (apoB)-depleted sera of healthy subjects (control, n = 27) and patients with age-related macular degeneration (AMD, n = 29) were examined for (A) their ability to promote [³H]-cholesterol efflux from macrophages. [³H]-cholesterol-labeled RAW264.7 macrophages were incubated with 2.8% apoB-depleted sera for 4 hours. Cholesterol efflux is expressed as radioactivity in the supernatant relative to total radioactivity (in supernatant and cells). Values shown represent means of two independent experiments.</p

Paraoxonase activity.

<p>Activity of HDL-associated paraoxonase was measured using phenylacetate as substrate. Paraoxonase activity of apoB-depleted sera was calculated from the slopes of the kinetic chart. Values shown represent means of four independent experiments.</p

HDL-apolipoproteins and HDL associated lipids.

<p>Levels of total cholesterol (A), non-esterified cholesterol (FC) (B), phospholipids (PL) (C), free fatty acids (FFA) (D), triglycerides (TG) (E) were measured enzymatically in apoB depleted serum. HDL associated apolipoproteins ApoA-I (F), apoA-II (G), apoC-II (H), apoC-III (I) and apoE (J) were determined in apoB-depleted serum by immunoturbidimetry.</p

Anti-inflammatory capacity.

<p>(A) U937 monocytes containing a reporter cassette for factor-κB (NF-κB) were pretreated in the absence and presence of 10% lipoprotein deficient sera (LPDS) in the presence of indicated concentrations of reconstituted HDL (rHDL). After 1 ½ hours, cells were stimulated with LPS (50 ng/ml) for 24 hours, followed by assessment of GFP expression by flow cytometry. (B) ApoB-depleted sera of healthy subjects and AMD patients were analyzed for their ability to inhibit lipopolysaccharide LPS-induced NF-κB activation in monocytes. U937 monocytes were pretreated with 7% apoB- depleted sera. After 1 ½ hours, cells were stimulated with LPS (50 ng/ml) for 24 hours, followed by assessment of GFP expression by means of flow cytometry. Values shown represent means of two independent experiments.</p

Anti-oxidative capability.

<p>The anti-oxidative activity of HDL was determined by inhibition of AAPH-initiated oxidation of the fluorescent dye dihydrorhodamine (DHR). Incubation of DHR in presence of apoB-depleted sera from healthy subjects or AMD patients led to a reduction in the oxidation of DHR. Values shown represent means of two independent experiments.</p

Lipoprotein associated phospholipase A2 (Lp-PLA2) activity.

<p>Lipoprotein associated phospholipase A2 (Lp-PLA2) activity of apoB-depleted sera was measured using 2-thio PAF as substrate. Lp-PLA2 activity of apoB-depleted sera was calculated from the slopes of the kinetic chart. Values shown represent means of two independent experiments.</p