Spectra obtained by MALDI-ToF MS analysis related to spots of vitamin D-binding protein (DBP).

<p>DBP C-terminal region spectrum (upper panel) and major coverage DBP spectrum (lower panel).</p

List of metabolites occurring in the <i>Rosmarinus officinalis</i> extract identified by LC MS/MS.

<p>Concentrations were determined by means of calibration curves with pure standards, as reported in material and methods (n. d. = not determined).</p><p>List of metabolites occurring in the <i>Rosmarinus officinalis</i> extract identified by LC MS/MS.</p

Levels of protein carbonyls.

<p>Protein carbonyls (PC) levels in the serum of control (CTR) and multiple sclerosis patients: remitting (REM) and relapsing (REL). Serum samples were assayed for PC by slot blot analysis. Error bars indicate SD for 6 samples per group. Densitometric values shown are given as raw values.</p

Summary of the proteins identified as differently expressed using the proteomics approach.

a<p>The p-value associated with fold-change calculated using a Student’s t-test.</p>b<p>The fold-change in spot density from three groups of matching: Rel vs Ctr; Rem vs Ctr; Rel vs Rem. The arrow indicates the direction of change.</p

List of metabolites occurring in the <i>Rosmarinus officinalis</i> extract identified by LC MS/MS.

<p>Concentrations were determined by means of calibration curves with pure standards, as reported in material and methods (n. d. = not determined).</p><p>List of metabolites occurring in the <i>Rosmarinus officinalis</i> extract identified by LC MS/MS.</p

Effect of <i>Rosmarinus officinalis</i> extract on A375 melanoma cells.

<p>(A) Metabolic activity (MTT test). (B) Cell viability (Trypan blue exclusion test). Data are expressed as % of cell survival with respect to control. Results are the mean ± SD from three independent experiments. * P ≤ 0.05 versus vehicle control.</p

Effect of apigenin (A), carnosol (B), luteolin (C), scutellarin (D) and rosmarinic acid (E) on metabolic activity of A375 melanoma cells, assayed by MTT assay.

<p>Data are expressed as % of cell survival with respect to control. Results are the mean ± SD from three independent experiments. * P ≤ 0.05 versus vehicle control.</p

2D protein expression maps.

<p>Proteomic profile of representative 2D-gels with proteins differently expressed in the three groups of matching: REL vs CTR (A), REM vs CTR (B) and REL vs REM (C). The identified proteins by mass spectrometry are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065184#pone-0065184-t002" target="_blank">Table 2</a>.</p

Involvement of Oxidative Stress in Occurrence of Relapses in Multiple Sclerosis: The Spectrum of Oxidatively Modified Serum Proteins Detected by Proteomics and Redox Proteomics Analysis

<div><p>Multiple sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system. Several evidences suggest that MS can be considered a multi-factorial disease in which both genetics and environmental factors are involved. Among proposed candidates, growing results support the involvement of oxidative stress (OS) in MS pathology. The aim of this study was to investigate the role of OS in event of exacerbations in MS on serum of relapsing-remitting (RR-MS) patients, either in relapsing or remitting phase, with respect to serum from healthy subjects. We applied proteomics and redox proteomics approaches to identify differently expressed and oxidatively modified proteins in the low-abundant serum protein fraction. Among differently expressed proteins ceruloplasmin, antithrombin III, clusterin, apolipoprotein E, and complement C3, were up-regulated in MS patients compared with healthy controls. Further by redox proteomics, vitamin D-binding protein showed a progressive trend of oxidation from remission to relapse, respect with controls. Similarly, the increase of oxidation of apolipoprotein A-IV confirmed that levels of OS are elevated with the progression of the disease. Our findings support the involvement of OS in MS and suggest that dysfunction of target proteins occurs upon oxidative damage and correlates with the pathology.</p></div

Summary of the proteins identified as differently oxidized using the redox proteomics approach.

a<p>The p-value associated with fold-change calculated using a Student’s t-test.</p>b<p>The fold-change in spot density from three groups of matching: Rel vs Ctr; Rem vs Ctr; Rel vs Rem. The arrow indicates the direction of change.</p