CD spectroscopy of peptide F3L1 indicates increased alpha-helical content.

<p>CD spectra of Peptide F and F3L1 were measured at 100μM concentration in water containing 1.25% trifluoroethanol.</p

ApoA-I-derived peptides used in this study.

<p>The alpha-helical regions in the lipid-free structure [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132780#pone.0132780.ref005" target="_blank">5</a>] of intact ApoA-I are indicated in italics. The centrally located proline residues in Regions C and D are indicated in bold.</p

CD spectroscopy of peptide F3L1 indicates increased alpha-helical content.

<p>CD spectra of Peptide F and F3L1 were measured at 100μM concentration in water containing 1.25% trifluoroethanol.</p

ApoA-I-derived peptides specifically inhibit binding of anti-ApoA-I IgG to immobilized ApoA-I.

<p>Competition ELISA to determine the capacity of peptides to block binding of anti-ApoA-I antibodies in plasma samples from three different patients known to be positive for anti-ApoA-I autoantibodies. Plasma samples were preincubated (2 h, room temperature) with peptides at the indicated concentrations prior to addition to assay wells. Percent maximal ELISA signals were calculated as 100 × ([signal in well]-[mean background signal (uncoated well)])/ ([mean maximal signal (no peptide)]-[mean background signal]). Results are expressed as mean ± SD (n = 3).</p

Predicitve accuracy for acute ischemic coronary events of immunoreactivity to F3L1 and to native ApoA-I.

<p>Predicitve accuracy for acute ischemic coronary events of immunoreactivity to F3L1 and to native ApoA-I.</p

The Human Autoantibody Response to Apolipoprotein A-I Is Focused on the C-Terminal Helix: A New Rationale for Diagnosis and Treatment of Cardiovascular Disease? - Table 2

<p>*P value was computed by comparing patients with high vs low anti-F3L1 immunoreactivity. CHD: coronary heart disease, bpm: beats per minute, GFR: glomerular filtration rate.</p><p>The Human Autoantibody Response to Apolipoprotein A-I Is Focused on the C-Terminal Helix: A New Rationale for Diagnosis and Treatment of Cardiovascular Disease? - Table 2 </p

The anti-ApoA-I autoantibody response is strongly biased towards the C-terminal alpha-helical region.

<p>ELISA experiments were carried out using a set of ApoA-I-derived peptides (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132780#pone.0132780.t001" target="_blank">Table 1</a>). a. Capture ELISA assay to determine the immunoreactivity of plasma pooled from patients known to be positive for anti-ApoA-I autoantibodies against the set of peptides. Specific ELISA signals were calculated as [signal in well]-[mean background signal (uncoated well)]. Results are expressed as mean ± SD (n = 3) b and c. Competition ELISA to determine the capacity of the set of peptides to block binding of anti-ApoA-I antibodies from either pooled patient plasma (b) or goat polyclonal IgG (c) to immobilized ApoA-I. Plasma or antibody was preincubated (2 h, room temperature) with peptides at the indicated concentrations prior to addition to assay wells. Percent maximal ELISA signals were calculated as 100 × ([signal in well]-[mean background signal (uncoated well)])/([mean maximal signal (no peptide)]-[mean background signal]). Results are expressed as mean ± SD (n = 3).</p

Peptide F3L1 dose-dependently inhibits anti-ApoA-I IgG-induced TNF-α and IL-6 production.

<p>Anti-ApoA-I IgG was incubated with the indicated F3L1 concentrations (preincubation 2 h at room temperature) prior to addition to cultured human monocyte-derived macrophages. Levels of proinflammatory cytokines were determined after 24 h culture. Experiments were repeated using cells from three different healthy donors, with results expressed as median, interquartile range (IQR) and range. Kruskal-Wallis test for a trend showed p value = 0.01 for TNF-α, and p value = 0.005 for IL-6.</p

Comparing the inhibitory potency of Peptide F and F3L1 on anti-ApoA-I IgG-induced TNF-α.

<p>Anti-ApoA-I IgG was incubated with the indicated peptide concentrations (preincubation 2 h at room temperature) prior to addition to cultured RAW cells. Levels of proinflammatory cytokines (TNF-α) were determined after 24 h culture. Mean levels (n = 3) are shown with error bars indicating the range. Kruskal-Wallis test for a trend showed p value = 0.01.</p

Illustrative cases.

<p>All MRI examinations were performed using a 1.5 Tesla scanner (Philips Medical System). Standard MRI work-up systematically comprised at least one series of T2-weighted images (turbo spin echo, repetition time msec (RT)/echo time msec (ET=0.625/120; numbers of signals averaged ((NAS)=2; turbo factor=15) and T1-weigthed images (spin echo, RT/ET= 500/10; NAS=2) obtained prior to and after gadolinium injection. Representative contrast-enhanced images from low (a,b,c) and high (d,e,f) levels of sVE patients. (A,B,C) 36 years old man, oligodendroglioma grade III, in the left posterior temporal region (6 cm major axis): (A) Sagittal T1-weighted image after gadolinium injection shows diffuse and extensive contrast enhancement, (B) axial T2-weighted image shows heterogeneous aspect and few perilesional edema of the same lesion with (C) T2/fluid attenuated inversion recovery (FLAIR) shows infiltrative lesion with mass effect on ventricular junction. On all panels, the tumor area is indicated using a dotted white line. For this patient, sVE=296 ng/mL and overall survival was 12 months. (D,E,F) 60 years old women, glioblastoma in the left parietal region (3.5 cm major axis): (D) Sagittal T1-weighted image after gadolinium injection shows a ring of contrast enhancement around an area of hypointensity (necrosis). (E) axial T2-weighted image (T2) shows irregular contours and significative perilesional edema. (F) T2/fluid attenuated inversion recovery (FLAIR) shows few mass effect. For this patient, sVE=1.843 µg/mL and overall survival was 36 months.</p