Additional file 1: of High-dose intravenously administered iron versus orally administered iron in blood donors with iron deficiency: study protocol for a randomised, controlled trial

SPIRIT 2013 Checklist: Recommended items to address in a clinical trial protocol and related documents*. (DOCX 63 kb

Additional file 9: of Vitamin D deficiency in critically ill children: a systematic review and meta-analysis

Vitamin D deficiency and clinical outcomes in the PICU. Figure showing association of vitamin D deficiency with mechanical ventilation. (TIF 74 kb

Additional file 1: of Vitamin D deficiency in critically ill children: a systematic review and meta-analysis

PRISMA checklist. (PDF 333 kb

Additional file 8: of Vitamin D deficiency in critically ill children: a systematic review and meta-analysis

Vitamin D deficiency and clinical outcomes in the PICU. Figure showing association of vitamin D deficiency with vasopressor use. (TIF 74 kb

Additional file 5: of Vitamin D deficiency in critically ill children: a systematic review and meta-analysis

Comparison of vitamin D status at PICU admission (or first day in PICU) of critically ill children and healthy control subjects. Table summarizing vitamin D status at PICU admission (or first day in PICU) of critically ill children and healthy control subjects, and the pooled mean difference in 25(OH)D levels between the PICU and control cohorts. (PDF 78 kb

Additional file 1: of Study protocol for a phase II dose evaluation randomized controlled trial of cholecalciferol in critically ill children with vitamin D deficiency (VITdAL-PICU study)

PIRIT checklist. Completed SPIRIT checklist. (PDF 60 kb

Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect

<div><p>We investigated a polyethylene glycol non-precipitable low-density lipoprotein (LDL) subfraction targeted by IgG and the influence of statin therapy on plasma levels of these small LDL-IgG-immune complexes (LDL-IgG-IC). LDL-subfractions were isolated from 6 atherosclerotic subjects and 3 healthy individuals utilizing iodixanol density gradient ultracentrifugation. Cholesterol, apoB and malondialdehyde (MDA) levels were determined in each fraction by enzymatic testing, dissociation-enhanced lanthanide fluorescence immunoassay and high-performance liquid chromatography, respectively. The levels of LDL-IgG-IC were quantified densitometrically following lipid electrophoresis, particle size distribution was assessed with dynamic light scattering and size exclusion chromatography. The influence of simvastatin (40 mg/day for three months) on small LDL-IgG-IC levels and their distribution among LDL-subfractions (salt gradient separation) were investigated in 11 patients with confirmed coronary artery disease (CAD). We demonstrate that the investigated LDL-IgG-IC are small particles present in atherosclerotic patients and healthy subjects. In vitro assembly of LDL-IgG-IC resulted in particle density shifts indicating a composition of one single molecule of IgG per LDL particle. Normalization on cholesterol levels revealed MDA values twice as high for LDL-subfractions rich in small LDL-IgG-IC if compared to dominant LDL-subfractions. Reactivity of affinity purified small LDL-IgG-IC to monoclonal antibody OB/04 indicates a high degree of modified apoB and oxidative modification. Simvastatin therapy studied in the CAD patients significantly lowered LDL levels and to an even higher extent, small LDL-IgG-IC levels without affecting their distribution. In conclusion simvastatin lowers levels of small LDL-IgG-IC more effectively than LDL-cholesterol and LDL-apoB levels in atherosclerotic patients. This antiatherogenic effect may additionally contribute to the known beneficial effects of this drug in the treatment of atherosclerosis.</p></div

Estimation of LDL-IgG-IC particle size.

<p>Protein staining (left panel; Ponceau staining) and IgG immunodetection (right panel) of LDL-subfractions after electrophoresis in 3% polyacrylamide slab gel (migration is possible for particles < 35 nm). The small LDL-IgG-IC particles of subfractions #11, #12 and #13 (IgG) show a similar mobility like the major LDL-subfraction #7 (protein stain) (<b>A</b>). High performance gel permeation chromatography (TSK 5000 PW column; 600 mm x 4 mm) of a small LDL-IgG-IC enriched LDL-subfraction. Fractions of 1 min in the elution region of LDL were collected (solid line: preparative isolation; sample = 80 μL of LDL-IgG-IC enriched subfraction). The dotted line represents an analytical run sample = 20 μL of LDL-IgG-IC enriched subfraction). The distribution of LDL-IgG-IC (preparative run; fractions of min 25–31) in the eluted fractions was visualized after dot-blot analysis (immunodetection of IgG) (insert) (<b>B</b>).</p

Simvastatin lowers small LDL-IgG-IC levels more effectively than cholesterol and apoB in patients with CAD.

<p>The reduction of total small LDL-IgG-IC levels is presented as percentage change from baseline for the 6 individual LDL-subfractions. Total amounts of small LDL-IgG-IC per fraction were calculated by conversion of the LDL-IgG-IC DELFIA counts. For each subject and each LDL fraction the baseline value and the value after statin therapy were used to calculate the difference as a percentage (post-statin minus pre-statin). Each bar represents the mean difference of total small LDL-IgG-IC per fraction on a percentage basis (*p < 0.05; **p < 0.01). (<b>A</b>). Comparison of the reduction of LDL-cholesterol, LDL-apoB levels and total small LDL-IgG-IC (average of reduction of LDL-subfractions) expressed as percentage change from baseline. Bars for LDL-cholesterol and LDL-apoB represent the percentage differences (post-statin minus pre-statin) determined in the entire LDL fractions. The reduction of the total amount of small LDL-IgG-IC of LDL is presented as average of the percentage differences calculated for LDL-subfractions 1–6 (shown in Fig 5A). Data represent means ± SD (<b>B</b>).</p

Identification of LDL-IgG-IC within the density range of LDL.

<p>Lipoproteins were separated by a self-generated iodixanol gradient single-step ultracentrifugation. Density of the obtained 25 subfractions and LDL region (<b>A</b>). Cholesterol levels of collected fractions from 3 healthy subjects (<b>B</b>). Distributions of cholesterol (solid lines), MDA (dotted lines) and LDL-IgG-IC detected by a specific anti-human-IgG antibody (insert at the top right) in patients with PAOD. MDA levels represent protein bound MDA (<b>C</b>).</p