Additional file

Title: A systematic examination of brain volumetric abnormalities in recent-onset schizophrenia using voxel-based, surface-based and region-of-interest-based morphometric analyses

Authors: John et al.

Table S1: Total number of subjects recruited into the study, reasons for dropping subjects, and final n of each sample.

Subject groups / Total number of subjects recruited into the study (N= 109) / Subjects dropped from analysis and reasons thereof / Number of subjects considered in the final analyses (N=90)
Healthy Control subjects / 51 / Loss of data due to hardware issues (computer hard disk crashed before back-up) / 5 / 45
Localizable brain pathology / 1
Schizophrenia subjects / 58 / Loss of data due to hardware issues (computer hard disk crashed before back-up) / 2 / 45
Poor image quality due to motion artefacts / 1
Exclusion following clarification of details subsequent to MRI acquisition, from reliable informants and from follow-up notes in the case files over 1-3 years after recruitment into the study (major reasons for dropping subjects: revision of diagnosis; comorbid conditions, especially substance dependence; longer duration of illness) / 10

Table S2: Details of lifetimeexposure to neurolepticsand duration of neuroleptic use (n=24).

Patient / Neuroleptic previously on (Dosage/duration) / Currently medicated with(Dosage/duration)
SZ_004 / No information* / Drug free
SZ_008 / Olanzapine 15mg*3mths, Aripiprazole 10mg*3 months,Trifluparazine20mg*1month,Amisulpiride 100mg*1month, / Clozapine 50mg*1month
SZ_009 / Olanzapine 20mg*4months / Drug free
SZ_012 / RSPN 4mg* 1week, Quetiapine 500mg*5weeks, RSPN 6mg*6months+2weeks / RSPN 4mg*3months
SZ_014 / No information* / Drug free
SZ_018 / Olanzapine 10mg*3years / Drug free
SZ_027 / No information* / Drug free
SZ_029 / No information*
SZ_031 / Olanzapine 5mg - 15mg*1 year 1month, RSPN 1mg-6mg*11months / RSPN 6mg*1month
SZ_032 / RSPN 6mg*5wks, Flupenthixol 3mg-6mg*3wks, Clozapine 100mg-400mg*3months 2wks / Drug free
SZ_034 / RSPN 4mg*3wks / RSPN 6mg*4.5months
SZ_036 / RSPN 4mg-8mg*7months 2wks / RSPN 6mg*2months
SZ_037 / RSPN 6mg-8mg*9 months / RSPN 6mg*1year
SZ_039 / RSPN 6mg*1month,Haloperidol 10mg*1month,Quetiapine 400mg-800mg*5months / Flupenthixol 3mg*1month
SZ_041 / RSPN 4mg-6mg*6months 1 wk, Flupenthixol 6mg*6wks,Amisulpiride 600mg-750mg*1year 5months,Haloperidol 10mg*4mths+2wks,Ziprasidone 150mg*1month 3 weeks / Ziprasidone 80mg*1week
SZ_042 / Olanzapine 10mg*3.5yrs,Levosulipride 100mg*2months / Levosulpiride200mg*1month
SZ_044 / RSPN 6mg*1year / RSPN4mg*4months
SZ_046 / No information* / Drug free
SZ_047 / Amisulpiride 400mg-750mg*9months 2weeks / RSPN4mg*7weeks
SZ_049 / No information* / Drug free
SZ_054 / No information* / Medicated; but no prescriptions available
SZ_055 / RSPN 6mg*2weeks / Drug free
SZ_058 / Olanzapine 15mg*9months,RSPN 4mg-8mg*9months,Flupenthixol 6mg*11months / Flupenthixol 3mg*3months
SZ_060 / RSPN 4mg-6mg*2month 2wks,Olanzapine 10mg-20mg*3months / Clozapine 150mg*4months

*missing values replaced by mean value in statistical analyses

Table S3:Comparison of total brain volume between males and females in the recent-onset schizophrenia (ROS) (N=45) and the healthy control subject (HCS) (N=45)samples. Between-gender analysis of covariance (ANCOVA) in both samples with age as covariate revealed significant gender differences in both samples when intracranial volume (ICV) was not used as an additional covariate; whereas there were no significant gender differences in total brain volume when ICV was used as an additional covariate.

Recent-onset schizophrenia (ROS) (N=45) / Healthy control subjects (HCS) (N=45)
Male
(N=32)
Mean (SD) / Females
(N=13)
Mean (SD) / Between-group ANCOVA without ICV as covariate / Between-group ANCOVA with ICV as covariate / Males
(N=35)
Mean (SD) / Females
(N=10)
Mean (SD) / Between-group ANCOVA without ICV as covariate / Between-group ANCOVA with ICV as covariate
1107.45 (114.53) / 987.31 (76.71) / F=14.46; p<0.00 / F=0.02; p<0.88 / 1131.65(106.71) / 958.47 (50.23) / F=22.14; p<0.00 / F=0.12; p<0.74

FigureS1: Freesurfer-generated difference (t) map of brain volumes depicting grey matter volume reduction in schizophrenia subjects (N=45) in comparison to healthy subjects (N=45) at p < 0.001 uncorrected with gender entered as fixed factor and
age, and total brain volume as nuisance factors.

Table S4: Brain regions showing volumetric reductions in schizophrenia subjectsin comparison to healthy subjects at a significance threshold of p<0.001 uncorrected, on vertex-wise analysis using Freesurfer, withgender entered as a fixed factor and age, and total brain volume as nuisance factors.

No. / Annotation / Max / VtxMax / Size(mm2) / TalX / TalY / TalZ
Left fusiform / 3.3850 / 91512 / 13.27 / -31.3 / -38.3 / -20.9
Right inferior temporal / 3.3850 / 91512 / 9.87 / 46.9 / -35.7 / -20.5
Left posterior cingulate / 3.1700 / 2869 / 22.05 / -6.4 / -17.7 / 39.1
Right supramarginal / 3.1700 / 2869 / 20.15 / 55.5 / -26.7 / 27.9
Left inferior temporal / 3.1564 / 101953 / 11.85 / -45.3 / -44.5 / -14.5
Right fusiform / 3.1564 / 101953 / 13.13 / 29.2 / -69.2 / -8.4
Left-gray matter on the banks of superior temporal sulcus / 3.0043 / 5131 / 0.43 / -58.4 / -52.1 / 2.3
Right pericalcarine / 3.0043 / 5131 / 0.42 / 19.8 / -71.9 / 8.9

Figure S2: Statistical parametric t-map of gray matter volumes shown as reduced in drug naïve schizophrenia subjects (N=21) in comparison to healthy control subjects (N=45) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels with total brain volume (TBV),age and gender were entered in the two sample random effects analysis (RFX) as co-variates.

Table S5: Brain regions showing volumetric reductions in drug naïve schizophrenia subjects (N=21) in comparison to healthy control subjects (N=45) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV),age and gender were entered in the two sample random effects analysis (RFX) as co-variates.

No. / Brain Areas / X coordinate / Y coordinate / Z coordinate / Z score
1 / Left Parahippocampal Gyrus / -34.15 / -11.21 / -13.58 / 3.967594
2 / Left Declive / -14.93 / -73.18 / -15.07 / 3.675226
3 / Left Inferior Occipital Gyrus * / -27.55 / -89.14 / -10.04 / 3.651867
4 / Left Middle Frontal Gyrus / -26.49 / 4.36 / 43.43 / 3.465131
5 / Right Declive / 18.45 / -82.49 / -22.14 / 3.382716
6 / Left Sub-Gyral / -20.37 / -17.52 / -7.19 / 3.309548
7 / Left Culmen / -20.36 / -46.21 / -16.66 / 3.246271
8 / Right Parahippocampal Gyrus * / 18.53 / -17.46 / -9.23 / 3.183492
9 / Right Superior Temporal Gyrus / 49.04 / -16.48 / -5.91 / 3.131638

*2 clusters significant at p<0.001 uncorrected; cluster with higher z-score listed

Figure S3: Statistical parametric t-map of gray matter volumes shown as reduced in medicated schizophrenia subjects (N=24) in comparison to healthy control subjects (N=45) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV),age and genderwere entered in the two sample random effects analysis (RFX) as co-variates.

Table S6: Brain regions showing volumetric reductions in medicated schizophrenia subjects (N=24) in comparison to healthy control subjects (N=45) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV),age and genderwere entered in the two sample random effects analysis (RFX) as co-variates.

No. / Brain Areas / X coor / Y coor / Z coor / Z-score
1 / Left Inferior Frontal Gyrus* / -38.33 / 38.08 / 1.83 / 3.896155912
2 / Left Middle Frontal Gyrus* / -39.58 / 44.59 / -7.03 / 3.548758875
3 / Left Frontal Sub-Gyral / -34.09 / 45.43 / -1.45 / 3.265842177
4 / Left Anterior Cingulate / -3.63 / 14.53 / -3.87 / 3.341869049
5 / Right Precentral Gyrus / 30.11 / -16.07 / 62.72 / 3.286299552
6 / Left Tuber / -35.61 / -77.49 / -27.99 / 3.186124323
7 / Left Superior Temporal Gyrus / -35.46 / 4.43 / -14.82 / 3.176743269
8 / Left Parahippocampal Gyrus* / -24.45 / -11.39 / -12.08 / 3.167867704
9 / Right Lingual Gyrus / 4.24 / -93.02 / -0.41 / 3.15545316

*2 clusters significant at p<0.001 uncorrected; cluster with higher z-score listed

Figure S4: Statistical parametric t-map of gray matter volumes shown as reduced in neuroleptic-naïve schizophrenia subjects (N=21) in comparison to medicated schizophrenia subjects (N=24) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV), age, genderand duration of illness were entered in the two sample random effects analysis (RFX) as co-variates.

Table S7: Brain regions showing volumetric reductions in neuroleptic-naïve schizophrenia subjects (N=21) in comparison to medicated schizophrenia subjects (N=24) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV), age, genderand duration of illness were entered in the two sample random effects analysis (RFX) as co-variates.

No. / Brain Areas / X coor / Y coor / Z coor / Z-score
1 / Right Posterior Declive / 12.86 / -75.86 / -17.56 / 3.9714
2 / Right Posterior Pyramis / 17.14 / -79.16 / -27.26 / 3.376048
3 / Right Inferior Parietal Lobule / 34.27 / -50.37 / 52.78 / 3.892291
4 / Left Posterior Declive / -3.76 / -74.12 / -20.37 / 3.185957

Figure S5: Statistical parametric t-map of gray matter volumes shown as increased in neuroleptic-naïve schizophrenia subjects (N=21) in comparison to medicated schizophrenia subjects (N=24) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV), age, gender and duration of illness were entered in the two sample random effects analysis (RFX) as co-variates.

Table S8: Brain regions showing volumetric increases in neuroleptic-naïve schizophrenia subjects (N=21) in comparison to medicated schizophrenia subjects (N=24) at a significance threshold of p<0.001 uncorrected and an extent threshold of 0 voxels when total brain volume (TBV), age, genderand duration of illness were entered in the two sample random effects analysis (RFX) as co-variates.

No. / Brain Areas / X coor / Y coor / Z coor / Z-score
1 / Right Precentral Gyrus / 21.78 / -22.88 / 60.58 / 3.102223
2 / Right Inferior Frontal Gyrus / 40.73 / 34.48 / 6.88 / 3.092246