Chronic CLA administration compromises short-medium term learning and memory in both uninjured and CCI-injured rats as assessed by latency to locate the Morris water maze (MWM) following novel platform placement.

<p>Second Run: Five minutes after Run 1 (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169494#pone.0169494.g004" target="_blank">Fig 4</a>), latency in seconds to reach the hidden platform at PID 6–10 (Acquisition Phase; platform in SE quadrant) and at PID’s 20–22 (Re-acquisition Phase; platform changed to NE quadrant, ‘Novel Platform Placement’) was tested. MWM Run 2 latency data (mean ± SEM) on PID’s 6–10 and 21–22 were analyzed from 29 rats as follows: Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9). Data were analyzed using 2-way repeated measures ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.05; a, b, c and d = differences between time and group, e = differences within groups between the acquisition (PID 10) and re-acquisition phases (PID 20)).</p

CCI, but not CLA, reduces body weight.

<p>Body weight (as a % of baseline weight) each day from PID -10 to PID 29 for Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9) groups. Data were analyzed using 2-way repeated measures ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.05; letters indicate differences between pre- and post-surgery body weights between groups across the experiment).</p

CLA does not restore CCI-induced decreases in circulating sex hormone concentrations.

<p>Plasma concentrations (mean ± SEM) of P₄, T, 11-DOC and corticosterone (ng/mL) at PID’s -10, 1, 11, 19 and 29 for Sham + saline (n = 5), Sham + CLA (n = 5), CCI + saline (n = 8) and CCI + CLA (n = 5) groups. Data were analyzed using 2-way repeated measures ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.05, letters indicate differences between post-injury days and treatments for T, 11-DOC and corticosterone; for P₄, results did not reach significance for main effect of post-injury day or treatment, p = 0.068).</p

CLA does not reduce gross lesion size following a CCI.

<p>Top: Representative figures for each group, illustrating lesion size 1-month post-CCI. Bottom: Lesion size depicted as a percentage of surface area damaged (y-axis) for rats in each surgery/treatment group (x-axis). Gross lesion size data (mean ± SEM) at PID 29 were analyzed from 29 rats as follows: Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9). Data were analyzed using ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.01; letter indicates differences between groups).</p

Chronic CLA administration is detrimental to medium-long term spatial memory.

<p><u>Probe Test 1:</u> At the end of each acquisition phase, the platform was removed (from the NE quadrant) and 48 h later the animals were tested in the MWM for NE quadrant preference over 60 s to determine if they remembered the location of the hidden platform. <u>Probe Test 2:</u> Likewise, after completing the re-acquisition phase the platform was removed from the SE quadrant and 48 h later the animals were tested for SE (new location), NE (old location) and SE + NE quadrant preferences (both new and old locations). MWM duration in target quadrant(s) data (mean ± SEM) during Probe Test 1 and 2 were analyzed from 29 rats as follows: Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9). Data were analyzed by ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.05; a, b, c and d = differences between time and group, e = differences within groups between the acquisition (PID 10) and re-acquisition phases (PID 20)).</p

CLA does not improve vestibulomotor function following CCI.

<p>Rats were placed on a rotating rod accelerating at a constant rate (1 rotation per second per second) and allowed to run for up to 300 s at PID’s -3, 2, 6, 9, 13 and 23. Time elapsed (seconds) before fall was recorded. Rotarod latency data (mean ± SEM) on PID’s 6–10 and 21–22 were analyzed from 29 rats as follows: Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9). Data were analyzed using 2-way repeated measures ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test; letters indicate differences between groups and times compared to baseline and sham groups, <i>p</i> < 0.05.</p

Chronic CLA administration compromises medium-long term learning and memory in both uninjured and CCI-injured rats as assessed by latency to locate the Morris water maze (MWM) platform.

<p>First Run: Latency in seconds to reach the hidden platform at PID’s 6–10 (Acquisition Phase; platform in SE quadrant) and at PID 20–22 (Re-acquisition Phase; platform changed to NE quadrant, ‘Novel Platform Placement’). MWM Run 1 latency data (mean ± SEM) on PID’s 6–10 and 21–22 were analyzed from 29 rats as follows: Sham + saline (n = 5), Sham + CLA (n = 7), CCI + saline (n = 8) and CCI + CLA (n = 9). Data were analyzed using 2-way repeated measures ANOVA; post-hoc analyses were performed using the Tukey multiple comparison test (p < 0.05; a, b, c and d = differences between time and group, e = differences within groups between the acquisition (PID 10) and re-acquisition phases (PID 20)).</p

Baseline characteristics of the women in the KEEPS-Cog.

<p>^a30-point scale; scores < 24/30 can suggest impairment.</p><p>^b21-item scale; score ranges from 0 to 63, and a higher score indicates more symptoms of depression. Women were excluded from participation if they obtained a score of >28 at screening or baseline, suggesting severe depression.</p><p>^cDefined as a binary variable (yes = 1; no = 0).</p><p>^d50-point, multiple-choice test of vocabulary that provided estimate of intelligence.</p><p>^e100-point scale; score < 85/100 can suggest impairment.</p><p>^fCognitive factor scores, produced by a factor analysis, have a metric similar to standardized scores. SDs, however, varied from factor to factor.</p><p>*<i>p</i> < 0.05.</p><p>GED, General Educational Development; HDL, high-density lipoprotein; HOMA-IR, homeostatic model assessment–insulin resistance; LDL, low-density lipoprotein; POMS, Profile of Mood States; SD, standard deviation.</p><p>Baseline characteristics of the women in the KEEPS-Cog.</p

CONSORT diagram.

<p>CONSORT diagram.</p

Factor model for the cognitive baseline data.

<p>CFI, comparative fit index; RMSEA, root mean square error of approximation; TLI, Tucker-Lewis index.</p