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1 Supplementary Online Content Hooshmand B, Magialasche F, Kalpouzos G, et al. Association of vitamin B, folate, and sulfur amino acids with brain magnetic resonance imaging measures in older adults: a longitudinal population-based study. JAMA Psychiatry. Published online April 7, 06. doi:0.00/jamapsychiatry eappendix. Methods etable. Associations of Vitamin B, Red Blood Cell Folate, and Sulfur Amino Acid Levels With Change in Gray Matter and White Matter Volume Over 6 Years etable. Associations of Vitamin B, Red Blood Cell Folate, and Sulfur Amino Acid Levels With Change in Cerebrospinal Fluid and Hippocampal Volumes Over 6 Years efigure. Flowchart of Study Participants This supplementary material has been provided by the authors to give readers additional information about their work.
2 eappendix. Methods Measurement of blood pressure After a rest period of five minutes, arterial blood pressure was measured twice at a 5-min interval in a seated position on the left arm with a sphygmomanometer, and each measure was recorded immediately,. The Pearson correlation coefficient between two measurements was 0.90, p<0.00. The mean of the two readings was calculated and used in the analyses. Vitamin supplements were coded following the Anatomical Therapeutic and Chemical classification system. Among those who were supplemented at baseline, the mean (standard deviation (SD)) of vitamin B was (36.) pmol/l and the mean (SD) of holotc was 5. (88.) pmol/l. Brain imaging cohort Of the baseline study population (n = 50), 333 participants belonged to the younger cohort (i.e., those who were <78y at baseline) and 68 participants belonged to the older cohort (i.e., those who were 78 y at baseline). Follow-up examinations were taken place at 6-year interval for younger cohorts and at 3-year intervals for the older cohorts, and the survey methods were similar to those applied at the baseline examination. Among the older cohort (i.e., those who were 78 at baseline, n=68), 55 subjects refused to undergo a new MRI scan or had contraindications, 3 subjects died, subjects moved and 4 subjects had poor MRI quality; therefore, 9 subjects had available MRI scan at 3-year follow-up. The second follow-up of the older cohorts and the first followup of the younger cohorts were carried out from 007 through 00. From the older cohort, 6 subjects refused to undergo a new MRI scan or had contraindications, 7 subjects died and 6 subjects had poor MRI quality; therefore, 53 participants had available MRI the scan at 6-year follow-up. Of the younger cohort (i.e. those < 78 y at baseline, n=333), 9 subjects refused to undergo a new MRI scan or had contraindications, 0 subjects died, subjects moved, and
3 subjects had poor MRI quality; therefore, MRI scans was available in 07 participants. In total, 99 subjects had MRI scans at either 3-year of 6-year follow-up. Brain imaging protocol The protocol included an axial 3D T-weighted fast field echo (FFE) [repetition time (TR) 5 ms, echo time (TE) 7 ms, flip angle (FA) 5, field of view (FOV) 40, 8 slices with slice thickness.5 mm and in-plane resolution mm, no gap, matrix 56 56], and an axial turbo FLAIR sequence (TR 6000 ms, TE 00 ms, inversion time 900 ms, FA 90, ETL, FOV 30, slices with slice thickness 5mm and in-plane resolution mm, gap mm, matrix 56 56) sequences. The light clean up option was used to further remove odd voxels from the segmentations. To assess the intra-rater reliability, a randomly selected subset of 30 FLAIR images underwent a second assessment, and Dice coefficient, which measures the extent of spatial overlap between binary images, was calculated. Mean Dice coefficient was 0.76, which represents a very good test-retest agreement 3. Results Subjects with incident dementia at follow-up were older at baseline [mean age (dtandard deviation (SD)): 80.0 (6.7) versus 70.4 (8.9) years], less educated [mean (SD): 9. (.8) versus.8 (4.5) years], had a lower MMSE score [mean (SD): 8. (.6) versus 9. (.)], and were more likely to have higher cysteine levels [339.0 (33.) versus 3.7 (50.9) μmol/l] compared with non-demented participants.
4 References. Chobanian AV, Bakris GL, Black HR, et al. Seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. Hypertension. Dec 003;4(6): Aronow WS, Fleg JL, Pepine CJ, et al. ACCF/AHA 0 expert consensus document on hypertension in the elderly: a report of the American College of Cardiology Foundation Task Force on Clinical Expert Consensus documents developed in collaboration with the American Academy of Neurology, American Geriatrics Society, American Society for Preventive Cardiology, American Society of Hypertension, American Society of Nephrology, Association of Black Cardiologists, and European Society of Hypertension. Journal of the American College of Cardiology. May 7 0;57(0): Simoes R, Monninghoff C, Dlugaj M, et al. Automatic segmentation of cerebral white matter hyperintensities using only 3D FLAIR images. Magnetic resonance imaging. Sep 03;3(7):8-89.
5 etable. Associations of Vitamin B, Red Blood Cell Folate, and Sulfur Amino Acid Levels With Change in Gray Matter and White Matter Volume Over 6 Years Gray Matter White Matter Volume Volume Folate Crosssectional 0.55 (0.0); (0.07); Vitamin time (0.0); (0.0); 0.59 Crosssectional (0.36); (0.43); 0.47 Vitamin time (0.0); (0.0); B 0.08 (0.06); (0.4); (0.00); (0.0); (0.4); (0.53); (0.00); (0.0); Holotrascobalamin 0.06 (0.06); (0.6); (0.00); (0.0); (0.4); (0.55); (0.00); (0.00); Homocysteine (0.08); (0.5); (0.); (0.0); (0.45); (0.55); (0.0); (0.0); 0.56 Methionine (0.09); (0.08); (0.00); (0.00); (0.45); (0.44); (0.00); (0.009); Cystathionine (0.059); (0.063); (0.005); (0.005); (0.079); (0.084); (0.005); (0.005); Cysteine (0.0); (0.7); (0.00); (0.0); (0.48); (0.57); (0.00); (0.00); Glutathione 0.5 (0.04); (0.00); (0.36); (0.00); (0.094); (0.009); (0.); (0.009); 0.85 β represents the coefficient for one standard deviation change in each compound and SE represents the standard error. Associations were examined by linear mixed models. The term cross-sectional represents the cross-sectional association between B, folate or sulfur amino-acids and brain volumes at baseline. The term Vitamin/sulfur amino-acid time represents the effect of B or folate or
6 sulfur amino-acids on the rate of change in brain volumes per year. A positive coefficient for Vitamin/Sulfur amino-acid time indicates that an increase in the vitamin/sulfur amino-acid was associated with a decreased rate of brain atrophy over time. Average yearly change without including vitamin/sulfur amino-acids in the model: (0 444), p<0 000 For cross sectional analysis: n=50 for folate, n= 470 for vitamin B, holotranscobalamin, and sulfur amino-acids 3 For longitudinal analysis, n=99 for those with available follow-up MRI scans and baseline folate s; n=8 for those with available follow-up MRI and available vitamin B, holotransciobalamin and sulfur amino-acids. : adjusted for age and sex and their interactions with time. : additionally adjusted for education, creatinine, mean systolic blood pressure, APOEε4 status, the use of vitamin supplements, smoking, treatment of hypertension, plasma cholesterol, obesity and their interactions with time.
7 etable. Associations of Vitamin B, Red Blood Cell Folate, and Sulfur Amino Acid Levels With Change in Cerebrospinal Fluid and Hippocampal Volumes Over 6 Years Cerebrospinal Hippocampal Fluid Volume Volume Folate Cross-sectional (0.36); (0.43); Vitamin time (0.05); (0.05); Cross-sectional (0.00); (0.00); 0.88 Vitamin time B (0.4); (0.5); (0.03); (0.04); (0.00); (0.00); Holotrascobalamin 0.35 (0.4); (0.55); (0.0); (0.04); (0.00); (0.00); Homocysteine (0.43); < (0.53); < (0.05); (0.06); (0.00); (0.003); Methionine (0.45); (0.45); (0.03); (0.0); (0.00); (0.00); Cystathionine 0.03 (0.079); (0.084); (0.007); (0.007); (0.00); (0.00); (0.000); (0.00); Cysteine 0.07 (0.48); (0.04); (0.00);
8 0.037 (0.57); (0.04); (0.00); Glutathione (0.30); (0.8); (0.03); (0.0); (0.00); (0.00); (0.000); β represents the coefficient for one standard deviation change in each compound and SE represents the standard error. Associations were examined by linear mixed models. The term cross-sectional represents the cross-sectional association between B, folate or sulfur amino-acids and brain volumes at baseline. The term Vitamin/sulfur amino-acid time represents the effect of B or folate or sulfur amino-acids on the rate of change in brain volumes per year. A positive coefficient for Vitamin/Sulfur amino-acid time indicates that an increase in the vitamin/sulfur amino-acid was associated with a decreased rate of brain atrophy over time. Average yearly change without including vitamin/sulfur amino-acids in the model: (0 444), p<0 000 For cross sectional analysis: n=50 for folate, n= 470 for vitamin B, holotranscobalamin, and sulfur amino-acids 3 For longitudinal analysis, n=99 for those with available follow-up MRI scans and baseline folate s; n=8 for those with available follow-up MRI and available vitamin B, holotransciobalamin and sulfur amino-acids. : adjusted for age and sex and their interactions with time. : additionally adjusted for education, creatinine, mean systolic blood pressure, APOEε4 status, the use of vitamin supplements, smoking, treatment of hypertension, plasma cholesterol, obesity and their interactions with time. For cross-sectional analysis: n=50 (CSF) and 49 (hippocampus) for folate, and n= 470 (CSF) and 457 (hippocampus) for vitamin B, holotranscobalamin, and sulfur amino-acids 3 For longitudinal analysis over 6 years: n=6 (CSF) and 55 (hippocampus) for folate, n=46 (CSF) and 39 (hippocampus) for vitamin B, folate, and sulfur amino-acids : adjusted for age and sex and their interactions with time. : additionally adjusted for education, creatinine, mean systolic blood pressure, APOEε4 status, the use of vitamin supplements, smoking, treatment of hypertension, plasma cholesterol, obesity, and their interactions with time.
9 efigure. Flowchart of Study Participants
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