5 T clinical scanner.

For fMRI, a BOLD sensitive sequence was used (echo time (TE) = 40 msec, repetition time (TR) = 2700 msec, flip angle = 90°). Thirty-two transversal PLX-4720 in vivo slices were acquired in interleaved fashion. The voxel size was 3 × 3 × 3 mm3. The number of dynamics for the complex working memory task was 302. The complex working memory task was presented to the participants using MR compatible video goggles (Resonance Technology Inc, Northridge, CA) and Superlab software (Cedrus Corporation, San Pedro, CA). The participants made their responses using a LUMItouch button box (Photon Control Inc., Burnaby, BC). fMRI analysis Image analysis was performed using SPM8 software (Wellcome Department of Imaging Inhibitors,research,lifescience,medical Neuroscience, Inhibitors,research,lifescience,medical University College, London, UK) applying the General linear model. Images in each fMRI scan were realigned to correct for movement during scanning and normalized to the Montreal Neurological Institute (MNI) template. Thereafter, the normalized images were smoothed with an 8 mm Gaussian kernel for noise reduction and to ameliorate differences

in intersubject localization. The images were analyzed using the standard parametric design to extract brain activation with increasing task difficulty during the word recollection phase. We assumed a linear BOLD response for increasing task Inhibitors,research,lifescience,medical difficulty using a contrast vector of −3 −1 1 3, which represents each block of word recollection as separate covariates, but with different weights according to the different difficulty levels determined by the number of sentences (1–4) presented before word recollection (levels 1–4). The presentation of sentences was modeled as a separate covariate Inhibitors,research,lifescience,medical but with zero

weight in the analysis. Contrast images of each participant were used in the second level analyses. Brain activation in the control group was assessed by a one-sample t-test of random effects and differences Inhibitors,research,lifescience,medical in brain activation between MS participants and controls were assessed by two-sample t-tests. In the two-sample t-tests, images from one group were exclusively masked by images check from the other group (mask P-value = 0.05, uncorrected). In this way, we obtained image maps of activation in one group that was not present in the other group. At the whole brain level of analysis, the resulting activation maps were significance thresholded at P = 0.001, uncorrected. Before the ROI analysis, we used a first level threshold of P = 0.01. Results were reported as significant if the cluster or peak P-value was less than 0.05, corrected for multiple comparisons using family wise error correction (FWE). In order to investigate if the brain activation in some of the predefined ROIs was correlated to perceived fatigue we used Fatigue VAS scores obtained after fMRI scanning as covariates in a second level analysis of all participants.