Markers’ change following a game confirms lack of correlation between S100B and UCH-L1.

<p><b>A–B</b>) refers to normalized serum surges of S100B and UCH-L1 after game 1 and game 2. Note lack of correlation between these markers (<i>p</i> = 0.19; R² = 0.37; game 1, <i>p</i> = 0.11; R² =  −0.45; game 2). The <i>red line</i> is the linear regression fit while the outer lines show confidence intervals of 95%. We measured the normalized percent change in each game in an attempt to correct for the possibility of different measuring sensitivities for subconcussive head hits for the two markers measured by ELISA. Significance is determined by ANOVA and denoted as (*) for p<0.05.</p

Parameters of Head Hit Index (HHI) calculations.

<p>A scoring system was used to segregate players based on the number and intensity of head hits experienced during games (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096296#s2" target="_blank">Methods</a> for details).</p

S100B serum surges correlate with the extent and number of head hits, while UCH-L1 does not appear to correlate with sub-concussive head hits.

<p>Mean percent change values of S100B and UCH-L1 levels (see equation (1)) were plotted against Head Hit Index (HHI) scores (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096296#s2" target="_blank">Methods</a>). A statistically significant difference (by Wilcoxon Mann Whitney) was found between S100B surges at HHI of 0 and HHI of 1 and 2. UCH-L1 did not correlate with any of the HHI used for this study, and the levels of UCH-L1 measured after the game and normalized for their pre-game value as in (1) did not discriminate between a HHI of 0 (special team or played without any head hit) and any HHI (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096296#pone.0096296-Marchi2" target="_blank">[10]</a> for details on HHI). Statistical differences were analyzed by the Wilcoxon Mann Whitney test, both in the comparison between S100B and UCH-L1 at a particular HHI and in the comparison of the individual marker at a HHI 0 vs. HHI 1, 2, 3, and 4,6. To correct for type 1 error in multiple comparisons the Dunnett’s correction was used. Significance is shown as (*) for p<0.05; and <i>n.s.</i> for not significant.</p

Lack of significant correlation between serum UCH-L1 levels and a diagnosis of mild TBI.

<p><b>A</b>) S100B levels are significantly elevated in patients with a diagnosis of mild TBI compared to healthy controls (<i>p</i><0.01) and, <b>B</b>) S100B levels correlate with post-traumatic findings on head CT (<i>p</i><0.01). <b>C</b>) <b>and D</b>) UCH-L1 levels remain unchanged in patients with a diagnosis of mTBI compared to controls as well as in those patients with positive findings on head CT. Significance is determined by ANOVA and denoted as (*) for p<0.05.</p

Comparison of SVID and metastases by MRI and age:

<p>A) Radiologic evaluation of SVID and metastases was based on comparison of post-contrast and FLAIR images. Note that metastases were obviously demarcated after gadolinium (Gd) injections, while SVID visible in FLAIR were not. The <i>red circles</i> refer to the locations of SVID or metastases in FLAIR or post-Gd images. B) Age distribution of patients affected by SVID or metastases. Patients with no metastases were younger than those with metastatic brain tumor; patients with SVID were significantly older than those without small vessel disease.</p

SVID grading methods, brain metastases identification, and metastatic distribution:

<p>A) Grading for SVID: deep white matter hyperintense signals, periventricular hyperintensity, and combined. Each represents the SVID distribution of grades of those with and without brain metastases. Differences were significant for deep white matter hyperintensity (p = 0.04), periventricular hyperintensity (p = 0.01), and the combined (p = 0.02). B) MRI image with gadolinium contrast demonstrates the protocol used to count identifiable metastases. These are indicated by empty red circles. C) Distribution of metastases in different CNS regions. Note that in the region where SVID are most common (cerebrum) there was a statistically significant difference in the number of metastases as predicted by a protective effect of SVID against tumor growth. See text for details.</p

Characteristics of lung cancer patients compared between those who presented with and those who presented without brain metastases.

<p>SD = standard deviation, SVID = small vessel ischemic disease.</p

Pattern of S100B, UCH-L1 and β-2 transferrin changes in players after football games.

<p>Serum samples were drawn as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096296#s2" target="_blank">Methods</a>. A total of 15 players were enrolled and analysis of samples reported here refers to two games played during the regular season. <b>A</b>)<b>, B</b>) and <b>C</b>) refer to absolute S100B, UCH-L1 and β-2 transferrin serum levels respectively. Serum levels were measured pre- (day before the game) and post-games (within one hour from the end of a game). In <b>D</b>) the normalized S100B, UCH-L1 and β-2 transferrin serum levels are shown side by side to allow a direct comparison. Normalized values for a given serum markers were obtained by the following equation: . In the figures, each symbol represents a player and any given player is represented by the same symbol throughout this manuscript. Note that on average S100B and UCH-L1 were increased after a game, while the values for beta-2 transferrin remained unchanged. Also note that the beta-2 transferrin values pre-game were highly variable (<b>C</b>) while most of baseline values for S100B and UCH-L1 fell within a defined range. Statistical differences by student's t-test are shown as (*) for p<0.05; and <i>n.s.</i> for not significant.</p

Relationship between SVID severity and metastatic brain tumor:

<p>The data are presented as % (filled symbols) or as a ratio between SVID severity in the two subsets of patients.</p

Serum levels of UCH-L1 do not correlate with S100B or levels of β-2 transferrin.

<p><b>A</b>) S100B serum levels measured at pre- and post- game did not correlate with UCH-L1 serum levels (<i>p</i> = 0.16; R² = 0.19). <b>B</b>) Lack of correlation between UCH-L1 or S100B and β-2 transferrin levels. β-2 transferrin serum levels measured at pre- and post- game (two games) plotted against UCH-L1 failed to show a statistically significant correlation (<i>p</i> = 0.28; R² = 0.20). <b>C</b>) S100B serum levels measured at pre- and post- game (two games) did not correlate with β-2 transferrin serum levels (<i>p</i> = 0.22; R² = 0.23). Significance is determined by ANOVA and denoted as (*) for p<0.05.</p