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Non-linear warping for the effect of spatial normalization was corrected to generate these modulated normalized images, which represent relative volume after correcting for brain size. Each image was then smoothed using an 8-mm full width at half maximum Gaussian kernel. A two-sample t-test was conducted for smoothed GM volume images from the ASD and TDC groups using a random-effect general linear model (GLM), with gender and age as nuisance regressors. Because the scans were taken at multiple sites which may have different MRI scanners and scanning protocols, an inherent variability may exist within the data. Therefore, we included an equal number of ASD and TDC participants within each site. We also used the locations as a dummy variable in our model. As suggested by the VBM8 manual, an absolute threshold mask of 0.1 was used for all the second-level analyses. To test the relationship between autism symptom severity and GM volume, we selleck products conducted an additional second-level GLM analysis for ASD participants, using their Autism Diagnostic Observation Schedule (ADOS) scores (Lord et al., 2000) as a regressor. Higher ADOS scores are indicative of increased ASD severity. Forty ASD participants for whom the ADOS scores were available (ABIDE n = 27; ISMMS n = 13) were selected from the original sample for this analysis. The significance level for the height of each voxel was set to p BKM120 datasheet with a contiguous-voxel extent threshold k > 17 voxels, to correct for multiple voxel comparisons. This threshold was estimated by using 10,000 Monte Carlo simulations with a customized Matlab program (Slotnick et al., 2003). The corrected a priori height threshold was p GUCY1B3 (Figure ?(Figure11 and Table ?Table2).2). In addition, compared to the TDC group, decreased GM volume in posterior brain regions in the ASD group was found, including the left posterior hippocampus and the cuneus bilaterally (Figure ?(Figure11 and Table ?Table2).2). These results remained consistent following an additional GLM analyses with age, gender, and site as nuisance regressors. Figure 1 Differences in gray-matter volume between the ASD and TDC groups. Red indicates areas of increased gray-matter volume in ASD (ASD > TDC). Blue indicates areas of decreased gray-matter volume in ASD (ASD