Histological analysis of the sciatic nerve 16 weeks post-PNI.

<p>For anatomical terminology, refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031675#pone-0031675-g001" target="_blank"><i>Fig. 1E</i></a>. (A) Myelination (MBP, green) of the regenerated axons (MAP2, red). Inset show the structure of individual myelinated axon fascicles, enlarged from 40× image. Axial sections 40×, longitudinal sections 20×. Thickness = 15 um. (B) Expression of laminin (green) was consistent with results from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031675#pone-0031675-g003" target="_blank"><i>Fig. 3A</i></a>. DAPI (blue), 20×. Thickness = 15 um.</p

Histological and quantitative analyses of macrophage presence.

<p>For anatomical terminology, refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031675#pone-0031675-g001" target="_blank"><i>Fig. 1E</i></a>. (A) ED1 IHC shows macrophage (red) location and number in each group in neuronal structure (NF-200, green). Note Sham, Wistar-DRG, NG, and Wistar Schwann represent macrophage presence in increasing order in all tissue sections. DAPI (blue), 20×. Thickness = 15 um. (B) Quantification and statistical analysis of ED1 immunoreactive cells. Macrophage count reflects immunohistochemistry microscopy analysis in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031675#pone-0031675-g004" target="_blank"><i>Fig. 4A</i></a>. ANOVA Bonferroni and Tukey multiple comparison post tests determined significant differences in the mean macrophage count in each group: *P<0.05; **P<0.01; ***P<0.001. Both post tests found the same results. Error bars represent ± standard error of the mean.</p

Evaluating the health of cultured neurons.

<p>(A) At the end of antimitotic treatment (day 3 and 4), the DRG cells (MAP-2 immunopositive neurons; green) had extended axons, and non-neuronal cells were essentially nonexistent. (B) Schwann cells demonstrated strong GFAP (red) expression and an elongated shape and dipolar morphology at day 2 and 3. 20×, Scale bar: 50 um. DAPI (blue).</p

MHC I immunoreactivity (red) in neuronal structure (NF-200, green).

<p>For anatomical terminology, refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031675#pone-0031675-g001" target="_blank"><i>Fig. 1E</i></a>. (A) Increased MHC I immunoreactivity were observed in NG, DRG, and Schwann groups compared to the Sham group. DAPI (blue), 20×. Thickness = 15 um. (B) Axon regeneration estimated by NF-200 expressing axons. Axon density in distal stump was calculated and expressed as standard optical density (SOD) for each group. n = 3, 40× image of 10 um sections.</p

Surgical Procedures.

<p>(A) After skin incision, an 8 mm length of sciatic nerve was excised. (B) A NeuraGen® nerve guide tube of 2.0 mm ID×1.0 cm length was used to provide a physical guide for axons sprouting from the proximal nerve stump to reach the disconnected nerve segment. (C) The rats were sacrificed at 16 weeks post-PNI and sciatic nerves were harvested by excising 1.0 cm outside the NG tube. (D) The tube (top) is distinguishable from the contralateral healthy tissue (bottom) as an enlarged section of the nerve. The contralateral nerve served as a control for each rat. (E) For histological analysis, frozen sections were cut and examined at five locations: nerve proximal (P) and distal (D) to the tube and proximal (Lp), distal (Ld), and middle (Lm) longitudinal regions inside the tube.</p

Diagnostic accuracy of S100B for sports-related concussion.

<p>A: The ROC using the 3-hour post-SRC value or the post-exertion S100B value revealed an AUC of 0.772 (95% CI: 0.64, 0.91). B: The AUC using the proportional increase in post-SRC/exertional S100B level over baseline was 0.904 (95% CI: 0.80, 1.0).</p

S100B Changes after non-contact exertion and sports-related concussion.

<p><b>A</b>: Absolute (Left) and proportional (right) changes in S100B levels after non-contact exertion among individal athletes. The proportional change is defined as (post-exertion or 3-hour post-SRC S100B – baseline S100B)/baseline S100B (range: −35% to +50%; mean proportional change  = +2.7%). The mean post-exertional S100B concentration (0.071±0.03 µg/L) was not significantly different from mean baseline (0.070±0.03 µg/L, *p = 0.87, Middle). <b>B</b>: Absolute (Left) and proportional (right) changes in S100B levels within 3 hours of SRC among individual athletes. The mean post-SRC S100B concentration within 3 hours of concussion (0.099±0.008 µg/L) was significantly higher than that at baseline (0.058±0.006 µg/L, **p = 0.0002, Middle). Proportional change (right) ranged from −14% to +180%; mean proportional change +81.2%). Munich athletes are designated in red, Rochester athletes in black.</p

Longitudinal changes in S100B before and after sports-related concussion.

<p><b>A</b>: Mean changes in S100B levels among all concussed athletes at baseline and at four time points after injury. The number of athletes included in determining the mean at each time point is indicated below each box plot. The mean S100B concentration within 3 hours of SRC (0.099±0.032 µg/L) was significantly higher than that at baseline (0.058±0.025 µg/L, *p<0.0001) using a paired t-test. The 3 hour S100B level was also higher than that at 2 days (0.059±0.03 µg/L, ⁺⁺p = 0.0004), 3 days (0.052±0.02 µg/L, ⁺p<0.0001), and 7 days (0.059±0.03 µg/L, **p = 0.001) post-SRC using unpaired t-tests. <b>B</b>: Longitudinal changes in S100B levels among concussed athletes who had baseline and all four post-SRC S100B measurements. All 11 of these athletes were from the Unviersity of Rochester. The mean S100B level within 3 hours of SRC (0.099±0.032 µg/L) was significantly higher than that at baseline (0.058±0.025 µg/L, *p<0.0001), and higher than that at 2 days (0.059±0.027 µg/L, ++p = 0.0047), 3 days (0.048±0.008 µg/L, +p = 0.0002), and 7 days (0.056±0.027 µg/L, **p = 0.0058) post-SRC using paired t-tests.</p

Concussion classification using S100B.

<p>Cutoff values shown are those that maximize sensitivity (†) and specificity (‡).</p

Subject characteristics.

<p>T-test to assess difference between Munich and Rochester cohorts at baseline, p = 0.85.</p