Restoration of learning ability by AD4 (100 mg/kg) 30 days post injury.

<p>Thirty days post injury, the mTBI mice showed a decline in cognitive performance both in <b>(A</b>) the Y-maze test and in the (<b>B</b>) NOR test (*p<0.001). Treatment with AD4 (100mg/kg) improved these impairments, revealed by LSD post hoc (p<0.001). Values are mean ± SEM.</p

Restoration of learning ability by AD4 (100 mg/kg) 30 days post injury.

<p>Thirty days post injury, the mTBI mice showed a decline in cognitive performance both in <b>(A</b>) the Y-maze test and in the (<b>B</b>) NOR test (*p<0.001). Treatment with AD4 (100mg/kg) improved these impairments, revealed by LSD post hoc (p<0.001). Values are mean ± SEM.</p

Thioredoxin-Mimetic-Peptides Protect Cognitive Function after Mild Traumatic Brain Injury (mTBI) - Fig 3

<p>The chemical structures of AD4 (top), TXM-CB3 (middle), and DY-70 (bottom).</p

Restoration of learning ability by TXM-CB3, AD4, and DY-70 (50mg/kg) 7 days post injury.

<p>Seven days post injury, the mTBI mice showed a decline in cognitive performance both in the (<b>A</b>) Y-maze test and in the (<b>B</b>) NOR test (***p<0.001, *p<0.05). Treatment with TXM-CB3 and DY-70 (50mg/kg) improved and AD4 did not improve the cognitive performance, revealed by LSD post. Values are mean ± SEM.</p

Administration of thiol reagnets alone did not affect leaning ability 30 days post injection.

<p>Mice administered AD4 and the TXM-peptides did not induce any learning deficit. All groups showed the same performance as the sham group <b>(A</b>) Spatial learning was tested using the Y-maze (N.S) and (<b>B</b>) Visual learning was tested using the NOR test (N.S). Values are mean ± SEM.</p

Restoration of learning ability by TXM-CB3, AD4, and DY-70 (50mg/kg) 30 days post injury.

<p>Thirty days post injury the mTBI mice showed a decline in cognitive performance both in (<b>A</b>) Y-maze test and in the (<b>B</b>) NOR test (***p<0.001). Treatment with TXM-CB3 (50 mg/kg) improved the cognitive performance. DY-70 (50mg/kg) did not improve the cognitive performance, revealed by LSD post. AD4 (50mg/kg) improved performance only in the Y maze test. Values are mean ± SEM.</p

Restoration of learning ability by AD4 (100mg/kg) 7 days post injury.

<p>Seven days post injury mTBI mice exhibit lower learning ability both (<b>A</b>) in the Y-maze test and in <b>(B</b>) the NOR test (*p<0.05). Treatment with AD4 (100mg/kg) improved this impairment, revealed by LSD post hoc (p<0. 01). Values are mean ± SEM.</p

Trx1 mimetic peptide TXM-CB13 (DY-70) reverses the auranofin-induced phosphorylation of JNK and p38^MAPK in SH-SH5Y cells.

<p>SH-SY5Y cells were treated with 7.5μM AuF for 30 min, washed, and treated with or without increasing concentrations of DY-70. Lysates were analyzed by immunoblotting with the indicated antibodies after separating proteins on SDS-PAGE. MAPK phosphorylation of (<b>A</b>) p38 (<b>B</b>) JNK or (<b>C</b>) ERK1/2 were visualized by immunoblot (<i>left</i>). The ratios of pp38, pJNK and ppERK1/2 to the housekeeping GAPDH protein were calculated based on three independent experiments (<i>right)</i>. The values shown are averages (±SEM) of three independent experiments normalized to the phosphorylation state of cells treated with AuF after 4.5hr. Student's <i>t</i>-test (two populations) was performed for AuF treated cells. *<i>P</i> value <0.05; **<i>P</i> value <0.01; ***<i>P</i> value < 0.005.</p

p53 inhibition by PFT-α/analog inhibits glutamate-induced excitotoxicity and oxidative stress mediated loss of cell viability in neuronal cultures.

<p>Human SH-SY5Y cells were subjected to p53 inactivation (PFT-α analog 1 to 10 μM) and then challenged with (<b>A</b>) glutamate (100 mM) excitotoxicity or (<b>B</b>) oxidative stress (H₂O₂: 500 μM). These insults alone significantly reduced cellular viability (* p<0.05 vs. control, Dunnetts t-test), which was mitigated by p53 inactivation (# p<0.05 vs. glutamate alone, Dunnetts t-test). (<b>C</b>) Rat primary cortical neuron cultures undergo time-dependent degeneration [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079837#B44" target="_blank">44</a>] that was mitigated by the addition of PFT-α (2 nM to 1 μM; * p<0.05, ** p<0.01, *** p<0.001 vs. untreated controls that are expressed as 100% (Dunnett’s t-test). A 10 μM PFT-α concentration proved to be toxic to primary neurons (*** p<0.001 vs. untreated controls; Dunnett’s t-test). (<b>D</b>) In an alike manner to SH-SY5Y cells, exposure of primary cortical neurons to glutamate (100 μM) resulted in reduced survival (* p<0.05 vs. control, Dunnetts t-test),) and pre-treatment with 2 to 100 nM PFT-α ameliorated this (NS not significantly different from untreated controls, Dunnetts t-test). Analysis of viable neurons was undertaken by MTS assay at 24 hr.</p

Time line of mouse studies.

<p>Anesthetized male ICR mice were subjected to either mTBI (a single 30 g weight drop) or a sham procedure (without weight drop) and 1 hr. later were administered either PFT-α (2 mg/kg, i.p.) or vehicle (0.2% DMSO/saline mixture, i.p.). Three parallel series of animals were then maintained for (i) 72 hr. and were prepared for immunohistochemical analyses of their brain tissue for quantification of degenerating neurons assessed by FJB and NeuN, (ii) 7 days and were behaviorally evaluated by novel object recognition and Y-maze paradigms, and (iii) 30 days and underwent similar behavior evaluation. The structure of PFT-α is shown as its synthesized HBr salt.</p