U.S. patent application number 13/626703 was filed with the patent office on 2013-05-02 for methods for the diagnosis of dementia and other neurological disorders.
This patent application is currently assigned to PHENOMENOME DISCOVERIES INC.. The applicant listed for this patent is PHENOMENOME DISCOVERIES INC.. Invention is credited to Lisa COOK, Dayan GOODENOWE.
Application Number | 20130110408 13/626703 |
Document ID | / |
Family ID | 38458611 |
Filed Date | 2013-05-02 |
United States Patent
Application |
20130110408 |
Kind Code |
A1 |
COOK; Lisa ; et al. |
May 2, 2013 |
METHODS FOR THE DIAGNOSIS OF DEMENTIA AND OTHER NEUROLOGICAL
DISORDERS
Abstract
The present invention is directed to a method for differentially
diagnosing dementia or the risk of dementia in a patient. The
method comprises obtaining a sample from the patient; analyzing the
sample to obtain quantifying data for one or more than one
metabolite marker; comparing the quantifying data for the one or
more than one metabolite marker to corresponding data obtained from
one or more than one reference sample; and using the comparison to
differentially diagnose dementia or the risk of dementia. The
method may also assist in assessing dementia or the risk of
dementia in a patient. The present invention is also directed to
metabolite markers and compounds useful in the present method.
Inventors: |
COOK; Lisa; (Saskatoon,
CA) ; GOODENOWE; Dayan; (Saskatoon, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
PHENOMENOME DISCOVERIES INC.; |
Saskatoon |
|
CA |
|
|
Assignee: |
PHENOMENOME DISCOVERIES
INC.
Saskatoon
CA
|
Family ID: |
38458611 |
Appl. No.: |
13/626703 |
Filed: |
September 25, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12280920 |
Feb 27, 2009 |
8304246 |
|
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PCT/CA2007/000313 |
Feb 28, 2007 |
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13626703 |
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60777290 |
Feb 28, 2006 |
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60804779 |
Jun 14, 2006 |
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60888883 |
Feb 8, 2007 |
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Current U.S.
Class: |
702/19 |
Current CPC
Class: |
C07F 9/106 20130101;
G01N 2800/52 20130101; G01N 2800/2814 20130101; G01N 2800/2821
20130101; G01N 33/6896 20130101; Y10T 436/163333 20150115; C07F
9/10 20130101; G01N 2800/50 20130101; G01N 30/7233 20130101; G16H
10/40 20180101 |
Class at
Publication: |
702/19 |
International
Class: |
G06F 19/00 20060101
G06F019/00 |
Claims
1. A method for diagnosing a patient's Alzheimer's disease health
state or change in health state, or for diagnosing Alzheimer's
disease, dementia, cognitive impairment, the risk of Alzheimer's
disease, the risk of cognitive impairment, or the risk of dementia
in a patient, the method comprising the steps of: a) analyzing a
blood sample from said patient by high resolution mass spectrometry
to obtain accurate mass intensity data; b) comparing the accurate
mass intensity data to corresponding data obtained from one or more
than one reference sample to identify an increase or decrease in
accurate mass intensity; and c) using said increase or decrease in
accurate mass intensity for diagnosing said patient's Alzheimer's
disease health state or change in health state, or for diagnosing
Alzheimer's disease, dementia, cognitive impairment, the risk of
Alzheimer's disease, the risk of cognitive impairment, or the risk
of dementia in said patient, wherein the accurate mass intensity is
measured, in Daltons, at or substantially equivalent to a hydrogen
and electron adjusted accurate mass, or neutral accurate mass,
selected from the group consisting of: 161.1053, 174.1409,
183.0661, 183.0662, 186.1408, 188.1566, 190.0330, 195.8577,
197.8549, 200.1564, 202.0453, 202.1720, 203.1157, 204.1876,
205.8865, 206.0789, 207.0822, 214.1720, 215.9153, 216.1877,
218.0192, 220.0798, 228.1877, 231.8893, 232.2188, 242.2030,
243.0718, 244.0559, 244.2187, 246.1465, 246.1472, 250.0366,
252.2095, 253.8163, 253.8164, 255.8135, 257.8105, 260.2135,
262.0221, 262.0777, 262.2293, 264.2452, 265.8423, 270.2343,
275.8712, 276.2095, 278.2254, 278.2255, 282.2573, 283.2602,
283.9028, 287.2824, 294.1443, 294.2207, 295.2239, 295.2287,
302.2255, 303.1079, 304.2402, 304.2410, 305.2438, 306.2569,
310.2883, 311.7750, 312.2312, 312.2663, 313.1153, 313.2690,
313.7721, 313.7722, 315.7693, 320.2356, 320.2357, 321.8039,
328.2402, 336.2662, 338.2821, 339.2850, 340.2976, 341.8614,
350.2423, 351.8903, 365.3159, 369.7338, 371.3542, 371.7311,
373.7280, 380.3096, 382.1082, 382.1083, 383.3283, 383.3284,
392.2938, 399.8196, 401.8166, 409.0208, 409.8484, 411.3211,
411.3212, 419.8773, 429.3749, 429.6888, 429.6890, 430.3818,
431.3860, 431.9570, 432.1532, 432.3254, 440.3532, 444.2717,
446.3403, 458.2404, 458.2405, 467.8070, 469.8042, 474.3706,
478.2540, 478.3664, 481.3172, 482.3215, 484.3041, 484.3794,
485.6503, 487.6482, 488.3873, 489.6451, 490.3641, 492.3816,
493.3850, 494.4342, 494.4343, 495.3320, 496.3355, 501.3212,
504.3814, 505.3229, 506.2851, 506.3213, 506.3214, 507.3316,
507.7055, 508.3356, 509.3493, 509.3494, 512.3356, 512.4082,
513.4122, 514.6879, 517.3136, 517.3140, 517.3722, 518.3174,
518.4345, 518.4346, 519.3321, 520.3353, 520.3354, 520.4499,
520.4500, 521.3474, 521.3477, 521.4522, 522.3511, 522.4634,
522.4635, 523.3634, 523.3635, 523.4678, 523.4679, 523.4680,
529.3166, 529.3167, 530.3213, 530.3471, 530.3820, 530.3821,
535.7944, 536.4794, 538.4224, 541.3139, 541.3430, 541.3432,
541.3433, 541.3435, 541.4425, 542.3173, 542.3461, 543.3296,
544.3970, 544.3971, 544.4479, 544.4481, 545.3453, 545.6062,
546.3485, 546.3486, 547.6031, 548.4815, 549.4844, 549.4845,
549.6005, 550.4956, 550.4957, 551.4985, 551.4986, 552.5021,
552.5022, 555.3098, 555.3101, 555.3102, 558.4667, 559.4690,
559.4700, 562.4600, 564.5134, 565.3393, 565.3394, 565.3396,
565.4104, 566.3434, 566.4130, 567.3545, 567.3547, 568.3573,
568.3574, 569.3685, 569.3687, 569.3690, 569.4784, 570.3724,
570.3725, 572.4468, 572.4816, 573.4852, 574.4597, 574.4637,
575.2728, 575.4632, 575.4990, 576.4770, 576.5097, 576.5098,
577.5136, 577.5137, 578.1574, 578.5277, 579.5325, 580.5350,
580.5351, 580.5352, 581.3344, 581.3345, 582.2473, 582.6334,
584.2646, 585.2664, 585.2673, 588.4731, 589.3403, 589.3404,
590.3430, 590.3431, 591.3542, 592.3571, 593.4743, 594.4878,
595.4932, 596.4791, 597.4840, 598.4965, 598.5124, 599.4936,
600.5127, 601.5164, 602.5282, 603.5317, 604.5432, 604.5433,
605.5456, 605.5457, 606.4869, 607.4919, 610.3686, 613.3405,
614.4914, 615.3539, 615.4798, 615.4938, 616.4673, 616.5052,
617.5089, 618.4829, 622.2539, 624.5131, 625.5075, 625.5076,
625.5161, 626.5104, 626.5108, 626.5277, 626.5278, 627.5201,
627.5302, 627.5304, 628.5237, 628.5421, 629.5449, 630.4861,
630.5586, 630.5587, 631.4902, 631.5619, 631.6280, 632.5032,
632.5035, 632.5762, 633.5078, 638.2465, 638.5149, 640.2637,
640.5294, 641.2661, 646.4837, 651.5221, 653.5360, 653.5361,
656.2587, 661.6233, 662.5175, 663.4870, 663.5216, 664.5322,
664.5323, 665.5010, 665.5354, 666.5455, 666.5456, 667.5474,
667.5475, 670.5689, 670.5690, 671.5722, 671.5723, 671.5726,
675.6357, 675.6377, 678.5477, 680.5623, 684.5489, 684.5491,
685.2600, 685.2601, 685.5538, 686.4877, 686.4880, 691.1957,
691.5960, 694.6161, 699.5198, 699.5199, 699.5200, 699.5203,
700.5520, 701.5359, 702.5677, 703.5516, 703.5709, 705.6093,
709.2594, 709.2595, 711.2577, 715.5167, 715.5169, 717.5308,
719.5465, 719.6231, 719.6233, 720.6258, 720.6438, 721.6390,
722.4789, 722.5334, 722.5335, 722.6424, 723.5194, 723.5195,
723.5197, 723.5203, 724.5256, 724.5257, 724.5258, 724.5493,
725.5359, 725.5385, 725.5527, 726.5459, 726.5461, 726.5462,
727.5516, 727.5567, 727.5568, 727.5569, 728.5626, 728.5627,
728.5629, 729.5672, 730.6493, 731.4914, 731.4916, 731.4917,
731.5463, 731.5464, 731.5829, 731.6530, 732.4937, 732.4938,
732.4939, 732.4940, 733.6426, 738.5445, 738.5448, 739.5141,
739.5142, 739.5143, 739.5164, 741.5305, 741.5307, 741.5319,
742.2972, 743.5464, 743.5466, 743.5467, 743.5468, 743.5469,
743.5470, 743.5471, 744.4954, 744.4956, 744.5500, 744.5535,
744.5536, 745.5621, 745.5656, 745.5658, 746.5119, 746.5570,
746.5714, 746.5717, 746.5719, 746.6210, 747.5201, 747.5203,
747.5244, 747.5245, 747.5778, 748.5286, 748.5287, 748.5721,
748.5735, 749.5359, 749.5365, 749.5367, 749.5406, 749.5407,
749.5408, 749.5760, 749.5761, 749.5777, 750.5402, 750.5403,
750.5438, 750.5440, 751.5516, 751.5529, 751.5553, 751.5555,
751.5556, 752.5564, 752.5581, 752.5583, 753.5672, 753.5672,
755.4860, 755.4861, 755.5467, 755.5468, 757.4989, 757.4991,
757.4993, 757.5014, 757.5016, 757.5625, 757.5626, 758.5092,
758.5094, 758.5655, 758.5656, 759.5162, 759.5163, 759.5164,
759.5165, 759.5779, 760.5215, 760.5216, 760.5217, 760.5811,
761.5843, 761.5846, 763.5157, 765.5313, 765.5334, 765.5335,
767.5469, 767.5470, 767.5494, 767.5495, 768.5503, 768.5504,
768.5538, 768.5539, 769.5654, 769.5655, 769.5656, 771.5813,
771.5814, 771.5815, 772.5278, 772.5280, 772.5860, 772.5862,
772.5863, 773.5347, 773.5368, 773.5370, 774.0313, 774.0316,
775.5516, 775.5527, 775.5528, 775.5530, 775.5533, 775.5535,
777.4426, 777.5287, 777.5288, 777.5530, 777.5531, 777.5672,
779.4393, 779.5441, 779.5442, 779.5444, 779.5829, 780.5474,
780.5475, 781.4354, 781.5607, 781.5616, 781.5617, 781.5620,
781.5985, 782.5084, 782.5085, 782.5086, 782.5647, 782.5649,
782.5650, 782.5653, 783.4315, 783.5147, 783.5148, 783.5671,
783.5672, 783.5778, 784.5235, 784.5237, 784.5239, 784.5810,
784.5811, 785.4288, 785.5933, 786.5415, 786.5416, 786.5967,
787.5464, 787.5465, 787.5728, 787.5729, 789.5892, 789.5893,
793.5386, 793.5387, 793.5679, 793.5681, 794.5421, 794.5422,
795.5181, 795.5182, 795.5183, 795.5550, 795.5551, 795.5837,
795.5838, 795.5839, 796.5290, 796.5292, 796.5874, 796.5876,
798.6742, 801.5550, 802.7056, 803.5445, 803.5677, 803.5678,
803.5680, 803.5681, 804.5475, 804.5476, 804.5713, 804.5718,
804.7227, 804.7230, 805.5604, 805.5605, 805.5831, 805.5832,
805.5839, 806.5637, 806.5638, 806.5639, 806.5873, 807.5757,
807.5758, 807.5768, 807.5899, 807.5900, 808.5792, 808.5803,
809.5936, 809.5937, 810.5399, 810.5400, 810.5401, 810.5969,
810.5971, 811.5732, 811.6096, 812.5761, 812.5762, 813.5885,
814.5917, 814.5919, 817.5375, 817.5376, 817.5377, 817.5378,
817.5838, 817.5840, 819.5551, 819.5552, 819.5553, 819.5641,
819.5642, 820.5677, 820.5679, 820.5680, 821.5711, 821.5712,
821.5713, 824.6892, 825.5544, 825.6926, 825.6927, 826.5581,
826.7048, 826.7069, 826.7070, 827.5448, 827.5699, 827.5700,
827.5701, 827.7083, 827.7086, 828.5736, 828.5737, 828.5742,
828.5743, 828.7202, 828.7207, 829.5604, 829.5856, 829.5859,
829.7239, 829.7242, 829.7244, 829.7246, 830.5894, 830.7352,
830.7355, 830.7362, 830.7363, 831.5759, 831.5760, 831.5995,
831.5997, 831.5998, 831.6000, 831.6001, 831.7387, 831.7408,
832.5791, 832.5792, 832.6024, 832.6026, 832.6027, 832.6036,
832.6037, 832.7492, 832.7495, 832.7521, 832.7522, 832.7523,
833.5931, 833.5932, 833.7551, 833.7558, 833.7570, 833.7571,
835.6998, 835.7001, 835.7006, 836.7076, 837.5027, 837.5881,
837.7180, 838.7226, 838.7232, 839.6031, 840.6063, 842.7387,
847.5316, 847.5953, 847.5954, 851.5698, 852.5738, 852.7250,
853.5854, 853.5855, 853.5862, 853.7296, 854.5902, 855.6015,
855.6016, 855.6023, 855.6025, 855.7417, 856.6061, 856.6063,
856.6720, 856.7481, 857.6186, 857.7531, 858.6211, 858.6212,
858.6860, 858.6861, 858.7632, 858.7663, 859.7694, 859.7695,
859.7696, 859.7706, 860.7752, 860.7753, 860.7756, 861.7801,
861.7806, 863.6876, 863.7336, 864.7380, 864.7596, 865.7486,
865.7487, 866.7532, 866.7533, 867.7579, 867.7581, 870.7307,
871.5527, 871.5528, 871.5529, 871.5934, 871.5935, 874.7645,
878.7400, 879.5999, 879.7454, 880.6035, 880.7516, 880.7566,
881.7558, 881.7616, 882.7660, 882.7723, 883.7705, 883.7765,
884.7801, 884.7877, 885.7854, 885.7922, 886.5582, 886.5584,
886.7916, 886.7917, 886.8012, 887.7352, 887.7999, 887.8001,
887.8005, 888.7394, 889.7492, 890.7535, 894.7810, 908.7832,
909.7882, 910.7968, 911.8032, 913.7513, 914.7583, 915.5191,
915.5192, 915.7681, 916.7743, 921.8142, 921.8145, 921.8153,
928.7505, 931.7695, 942.7879, 1018.9420, 1019.3838, 1019.3840,
1085.3294, 1098.9739, 1098.9740, 1225.0920, 1226.0968, 1226.0970,
1227.1070, 1228.1110, 1228.1111 and combinations thereof.
2. The method of claim 1, wherein the accurate mass intensity data
is obtained using a Fourier transform ion cyclotron resonance, time
of flight, orbitrap, quadrupole or triple quadrupole mass
spectrometer.
3. The method of claim 1, wherein the blood sample is a blood serum
sample.
4. The method of claim 1, wherein the accurate mass intensities
represent ionized metabolites.
5. The method of claim 1, wherein a liquid/liquid extraction is
performed on the blood sample whereby non-polar metabolites are
dissolved in an organic solvent and polar metabolites are dissolved
in an aqueous solvent.
6. The method of claim 5, wherein the accurate mass intensities are
obtained from the ionization of the extracted samples using an
ionization method selected from the group consisting of: positive
electrospray ionization, negative electrospray ionization, positive
atmospheric pressure chemical ionization, negative atmospheric
pressure chemical ionization, and combinations thereof.
7. The method of claim 1, wherein the accurate mass intensity data
is obtained using a Fourier transform ion cyclotron resonance mass
spectrometer.
8. The method of claim 1, wherein said one or more than one
reference sample is: a) one or more than one reference sample
obtained from a non-demented control individual; b) one or more
than one reference sample obtained from a demented subject; c) one
or more than one reference sample obtained from a subject
pathologically diagnosed as having Alzheimer's Disease; d) one or
more than one reference sample obtained from a subject clinically
diagnosed as having Alzheimer's Disease; e) one or more than one
reference sample obtained from a patient with cognitive impairment
as measured by Alzheimer's Disease Assessment Scale-cognitive
subset (ADAS-cog); f) one or more than one reference sample
obtained from a patient with cognitive impairment as measured by
Folstein's Mini-Mental State Exam (MMSE); or g) any combination of
a) to f) above.
9. The method of claim 1, wherein the method is for diagnosing: (a)
the presence of or risk of Alzheimer's disease (AD); (b) the
presence of or risk of AD pathology; (c) the presence of or risk of
dementia; (d) the presence of or risk of cognitive impairment; or
(e) a combination of (a) to (d) above.
10. The method of claim 1, further comprising analyzing a blood
sample from said patient by mass spectrometry to obtain accurate
mass intensity data for one or more than one internal control
metabolite; and calculating a ratio for each of the accurate mass
intensities obtained in step (a) to the accurate mass intensities
obtained for the one or more than one internal control metabolite;
wherein the comparing step (b) comprises comparing each ratio to
one or more corresponding ratios obtained for one or more than one
reference sample.
11. The method of claim 1, wherein a decrease in accurate mass
intensity is identified in the comparing step (b).
12. The method of claim 1, wherein the hydrogen and electron
adjusted accurate mass, or neutral accurate mass, is selected from
the group consisting of: 699.5198, 723.5195, 723.5197, 751.5555,
541.3432, 569.3687, 803.568, 886.5582, 565.3394, 569.369, 801.555,
857.6186, 207.0822, 275.8712, 371.7311, 373.728, 432.1532,
485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493, 742.2972,
701.53591, 699.52026, 723.52026, 747.52026, 729.56721, 727.55156,
779.58286, 775.55156, and combinations thereof.
13. A method for diagnosing a patient's Alzheimer's disease health
state or change in health state, or for diagnosing Alzheimer's
disease, dementia, cognitive impairment, the risk of Alzheimer's
disease, the risk of cognitive impairment, or the risk of dementia
in a patient, the method comprising the steps of: a) analyzing a
blood sample from said patient to obtain quantifying data for one
or more than one metabolite marker; b) comparing the quantifying
data for said one or more than one metabolite marker to
corresponding data obtained for one or more than one reference
sample to identify an increase or decrease in the level of said one
or more than one metabolite marker in said blood sample; and c)
using said increase or decrease in the level of said one or more
than one metabolite marker in said blood sample for diagnosing said
patient's Alzheimer's disease health state or change in health
state, or for diagnosing Alzheimer's disease, dementia, cognitive
impairment, the risk of Alzheimer's disease, the risk of cognitive
impairment, or the risk of dementia in said patient, wherein the
one or more than one metabolite marker comprises one or more
molecule selected from the group consisting of:
phosphatidylethanolamine (PtdEt), plasmenylethanolamine,
plasmanylethanolamine, and combinations thereof, wherein the PtdEt
is selected from the group consisting of PtdEt 16:0/18:0, PtdEt
16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, and combinations
thereof.
14. The method of claim 13, wherein the-plasmanylethanolamine is
selected from the group consisting of: plasmanyl 16:0/18:1,
plasmanyl 16:0/18:2, plasmanyl 16:0/20:4, plasmanyl 16:0/22:4,
plasmanyl 16:0/22:6, plasmanyl 18:0/18:1, plasmanyl 18:0/18:2,
plasmanyl 18:0/20:4, plasmanyl 18:0/22:4, plasmanyl 18:0/22:6, and
combinations thereof, and the plasmenylethanolamine is selected
from the group consisting of: plasmenyl 16:0/18:1, plasmenyl
16:0/18:2, plasmenyl 16:0/20:4, plasmenyl 16:0/22:4, plasmenyl
16:0/22:6, plasmenyl 18:0/18:1, plasmenyl 18:0/18:2, plasmenyl
18:0/20:4, plasmenyl 18:0/22:4, plasmenyl 18:0/22:6, and
combinations thereof.
15. The method of claim 13, wherein the quantifying data is
obtained using a Fourier transform ion cyclotron resonance, time of
flight, orbitrap, quadrupole or triple quadrupole mass
spectrometer.
16. The method of claim 15, wherein the mass spectrometer is
equipped with a chromatographic system.
17. The method of claim 13, wherein the blood sample is a blood
serum sample.
18. The method of claim 13, wherein a liquid/liquid extraction is
performed on the blood sample whereby non-polar metabolites are
dissolved in an organic solvent and polar metabolites are dissolved
in an aqueous solvent.
19. The method of claim 18, wherein the extracted samples are
analyzed by positive or negative electrospray ionization or
positive or negative atmospheric pressure chemical ionization.
20. The method of claim 18, wherein the extracted samples are
analyzed by MS/MS transition.
21. The method of claim 18, wherein the extracted samples are
analyzed by extracted ion current (EIC) chromatography and MS/MS
transition.
22. The method of claim 13, wherein said one or more than one
reference sample is a) one or more than one reference sample
obtained from a non-demented control individual; b) one or more
than one reference sample obtained from a demented subject; c) one
or more than one reference sample obtained from a subject
pathologically diagnosed as having Alzheimer's Disease; d) one or
more than one reference sample obtained from a subject clinically
diagnosed as having Alzheimer's Disease; e) one or more than one
reference sample obtained from a patient with cognitive impairment
as measured by Alzheimer's Disease Assessment Scale-cognitive
subset (ADAS-cog); f) one or more than one reference sample
obtained from a patient with cognitive impairment as measured by
Folstein's Mini-Mental State Exam (MMSE); or g) any combination of
a) to f) above.
23. The method of claim 13, further comprising: analyzing a blood
sample from said patient to obtain quantifying data for one or more
than one internal control metabolite; and obtaining a ratio for
each of the levels of said one or more than one metabolite marker
to the level obtained for the one or more than one internal control
metabolite; wherein the comparing step (b) comprises comparing each
ratio to one or more corresponding ratios obtained for the one or
more than one reference sample.
24. The method of claim 13, wherein a decrease in the level of said
one or more than one metabolite marker in said blood sample is
identified in the comparing step (b).
25. The method of claim 14, wherein the extracted samples are
analyzed by MS/MS transitions, and the MS/MS transitions for the
phosphatidylethanolamines are 718.0/255.0, 716.0/255.0, 746.0/283.0
and 744.0/283.0, respectively; the MS/MS transitions for the
plasmanylethanolamines are 702.0/281.0, 700.0/279.0, 724.0/303.0,
752.0/331.0, 748.0/327.0, 730.0/281.0, 728.0/279.0, 752.0/303.0,
780.0/331.0 and 776.0/327.0, respectively; and the MS/MS
transitions for the plasmenylethanolamines are 700.0/281.0,
698.0/279.0, 722.0/303.0, 750.0/331.0, 746.0/327.0, 728.0/281.0,
726.0/279.0, 750.6/303.2, 778.0/331.0 and 774.0/327.0,
respectively.
26. A method for evaluating the efficacy of a therapy for treating
Alzheimer's disease, dementia, or cognitive impairment in a
patient, comprising the steps of: a) analyzing a blood sample from
said patient by high resolution mass spectrometry to obtain
accurate mass intensity data; b) comparing the accurate mass
intensity data to corresponding data obtained from one or more than
one reference sample to identify an increase or decrease in
accurate mass intensity; and c) using said increase or decrease in
accurate mass intensity to determine whether the therapy is
improving the biochemical state of the patient, wherein the
accurate mass intensity is measured, in Daltons, at or
substantially equivalent to a hydrogen and electron adjusted
accurate mass, or neutral accurate mass, selected from the group
consisting of: 161.1053, 174.1409, 183.0661, 183.0662, 186.1408,
188.1566, 190.0330, 195.8577, 197.8549, 200.1564, 202.0453,
202.1720, 203.1157, 204.1876, 205.8865, 206.0789, 207.0822,
214.1720, 215.9153, 216.1877, 218.0192, 220.0798, 228.1877,
231.8893, 232.2188, 242.2030, 243.0718, 244.0559, 244.2187,
246.1465, 246.1472, 250.0366, 252.2095, 253.8163, 253.8164,
255.8135, 257.8105, 260.2135, 262.0221, 262.0777, 262.2293,
264.2452, 265.8423, 270.2343, 275.8712, 276.2095, 278.2254,
278.2255, 282.2573, 283.2602, 283.9028, 287.2824, 294.1443,
294.2207, 295.2239, 295.2287, 302.2255, 303.1079, 304.2402,
304.2410, 305.2438, 306.2569, 310.2883, 311.7750, 312.2312,
312.2663, 313.1153, 313.2690, 313.7721, 313.7722, 315.7693,
320.2356, 320.2357, 321.8039, 328.2402, 336.2662, 338.2821,
339.2850, 340.2976, 341.8614, 350.2423, 351.8903, 365.3159,
369.7338, 371.3542, 371.7311, 373.7280, 380.3096, 382.1082,
382.1083, 383.3283, 383.3284, 392.2938, 399.8196, 401.8166,
409.0208, 409.8484, 411.3211, 411.3212, 419.8773, 429.3749,
429.6888, 429.6890, 430.3818, 431.3860, 431.9570, 432.1532,
432.3254, 440.3532, 444.2717, 446.3403, 458.2404, 458.2405,
467.8070, 469.8042, 474.3706, 478.2540, 478.3664, 481.3172,
482.3215, 484.3041, 484.3794, 485.6503, 487.6482, 488.3873,
489.6451, 490.3641, 492.3816, 493.3850, 494.4342, 494.4343,
495.3320, 496.3355, 501.3212, 504.3814, 505.3229, 506.2851,
506.3213, 506.3214, 507.3316, 507.7055, 508.3356, 509.3493,
509.3494, 512.3356, 512.4082, 513.4122, 514.6879, 517.3136,
517.3140, 517.3722, 518.3174, 518.4345, 518.4346, 519.3321,
520.3353, 520.3354, 520.4499, 520.4500, 521.3474, 521.3477,
521.4522, 522.3511, 522.4634, 522.4635, 523.3634, 523.3635,
523.4678, 523.4679, 523.4680, 529.3166, 529.3167, 530.3213,
530.3471, 530.3820, 530.3821, 535.7944, 536.4794, 538.4224,
541.3139, 541.3430, 541.3432, 541.3433, 541.3435, 541.4425,
542.3173, 542.3461, 543.3296, 544.3970, 544.3971, 544.4479,
544.4481, 545.3453, 545.6062, 546.3485, 546.3486, 547.6031,
548.4815, 549.4844, 549.4845, 549.6005, 550.4956, 550.4957,
551.4985, 551.4986, 552.5021, 552.5022, 555.3098, 555.3101,
555.3102, 558.4667, 559.4690, 559.4700, 562.4600, 564.5134,
565.3393, 565.3394, 565.3396, 565.4104, 566.3434, 566.4130,
567.3545, 567.3547, 568.3573, 568.3574, 569.3685, 569.3687,
569.3690, 569.4784, 570.3724, 570.3725, 572.4468, 572.4816,
573.4852, 574.4597, 574.4637, 575.2728, 575.4632, 575.4990,
576.4770, 576.5097, 576.5098, 577.5136, 577.5137, 578.1574,
578.5277, 579.5325, 580.5350, 580.5351, 580.5352, 581.3344,
581.3345, 582.2473, 582.6334, 584.2646, 585.2664, 585.2673,
588.4731, 589.3403, 589.3404, 590.3430, 590.3431, 591.3542,
592.3571, 593.4743, 594.4878, 595.4932, 596.4791, 597.4840,
598.4965, 598.5124, 599.4936, 600.5127, 601.5164, 602.5282,
603.5317, 604.5432, 604.5433, 605.5456, 605.5457, 606.4869,
607.4919, 610.3686, 613.3405, 614.4914, 615.3539, 615.4798,
615.4938, 616.4673, 616.5052, 617.5089, 618.4829, 622.2539,
624.5131, 625.5075, 625.5076, 625.5161, 626.5104, 626.5108,
626.5277, 626.5278, 627.5201, 627.5302, 627.5304, 628.5237,
628.5421, 629.5449, 630.4861, 630.5586, 630.5587, 631.4902,
631.5619, 631.6280, 632.5032, 632.5035, 632.5762, 633.5078,
638.2465, 638.5149, 640.2637, 640.5294, 641.2661, 646.4837,
651.5221, 653.5360, 653.5361, 656.2587, 661.6233, 662.5175,
663.4870, 663.5216, 664.5322, 664.5323, 665.5010, 665.5354,
666.5455, 666.5456, 667.5474, 667.5475, 670.5689, 670.5690,
671.5722, 671.5723, 671.5726, 675.6357, 675.6377, 678.5477,
680.5623, 684.5489, 684.5491, 685.2600, 685.2601, 685.5538,
686.4877, 686.4880, 691.1957, 691.5960, 694.6161, 699.5198,
699.5199, 699.5200, 699.5203, 700.5520, 701.5359, 702.5677,
703.5516, 703.5709, 705.6093, 709.2594, 709.2595, 711.2577,
715.5167, 715.5169, 717.5308, 719.5465, 719.6231, 719.6233,
720.6258, 720.6438, 721.6390, 722.4789, 722.5334, 722.5335,
722.6424, 723.5194, 723.5195, 723.5197, 723.5203, 724.5256,
724.5257, 724.5258, 724.5493, 725.5359, 725.5385, 725.5527,
726.5459, 726.5461, 726.5462, 727.5516, 727.5567, 727.5568,
727.5569, 728.5626, 728.5627, 728.5629, 729.5672, 730.6493,
731.4914, 731.4916, 731.4917, 731.5463, 731.5464, 731.5829,
731.6530, 732.4937, 732.4938, 732.4939, 732.4940, 733.6426,
738.5445, 738.5448, 739.5141, 739.5142, 739.5143, 739.5164,
741.5305, 741.5307, 741.5319, 742.2972, 743.5464, 743.5466,
743.5467, 743.5468, 743.5469, 743.5470, 743.5471, 744.4954,
744.4956, 744.5500, 744.5535, 744.5536, 745.5621, 745.5656,
745.5658, 746.5119, 746.5570, 746.5714, 746.5717, 746.5719,
746.6210, 747.5201, 747.5203, 747.5244, 747.5245, 747.5778,
748.5286, 748.5287, 748.5721, 748.5735, 749.5359, 749.5365,
749.5367, 749.5406, 749.5407, 749.5408, 749.5760, 749.5761,
749.5777, 750.5402, 750.5403, 750.5438, 750.5440, 751.5516,
751.5529, 751.5553, 751.5555, 751.5556, 752.5564, 752.5581,
752.5583, 753.5672, 753.5672, 755.4860, 755.4861, 755.5467,
755.5468, 757.4989, 757.4991, 757.4993, 757.5014, 757.5016,
757.5625, 757.5626, 758.5092, 758.5094, 758.5655, 758.5656,
759.5162, 759.5163, 759.5164, 759.5165, 759.5779, 760.5215,
760.5216, 760.5217, 760.5811, 761.5843, 761.5846, 763.5157,
765.5313, 765.5334, 765.5335, 767.5469, 767.5470, 767.5494,
767.5495, 768.5503, 768.5504, 768.5538, 768.5539, 769.5654,
769.5655, 769.5656, 771.5813, 771.5814, 771.5815, 772.5278,
772.5280, 772.5860, 772.5862, 772.5863, 773.5347, 773.5368,
773.5370, 774.0313, 774.0316, 775.5516, 775.5527, 775.5528,
775.5530, 775.5533, 775.5535, 777.4426, 777.5287, 777.5288,
777.5530, 777.5531, 777.5672, 779.4393, 779.5441, 779.5442,
779.5444, 779.5829, 780.5474, 780.5475, 781.4354, 781.5607,
781.5616, 781.5617, 781.5620, 781.5985, 782.5084, 782.5085,
782.5086, 782.5647, 782.5649, 782.5650, 782.5653, 783.4315,
783.5147, 783.5148, 783.5671, 783.5672, 783.5778, 784.5235,
784.5237, 784.5239, 784.5810, 784.5811, 785.4288, 785.5933,
786.5415, 786.5416, 786.5967, 787.5464, 787.5465, 787.5728,
787.5729, 789.5892, 789.5893, 791.5386, 793.5387, 791.5679,
793.5681, 794.5421, 794.5422, 795.5181, 795.5182, 795.5183,
795.5550, 795.5551, 795.5837, 795.5838, 795.5839, 796.5290,
796.5292, 796.5874, 796.5876, 798.6742, 801.5550, 802.7056,
803.5445, 803.5677, 803.5678, 803.5680, 803.5681, 804.5475,
804.5476, 804.5713, 804.5718, 804.7227, 804.7230, 805.5604,
805.5605, 805.5831, 805.5832, 805.5839, 806.5637, 806.5638,
806.5639, 806.5873, 807.5757, 807.5758, 807.5768, 807.5899,
807.5900, 808.5792, 808.5803, 809.5936, 809.5937, 810.5399,
810.5400, 810.5401, 810.5969, 810.5971, 811.5732, 811.6096,
812.5761, 812.5762, 813.5885, 814.5917, 814.5919, 817.5375,
817.5376, 817.5377, 817.5378, 817.5838, 817.5840, 819.5551,
819.5552, 819.5553, 819.5641, 819.5642, 820.5677, 820.5679,
820.5680, 821.5711, 821.5712, 821.5713, 824.6892, 825.5544,
825.6926, 825.6927, 826.5581, 826.7048, 826.7069, 826.7070,
827.5448, 827.5699, 827.5700, 827.5701, 827.7083, 827.7086,
828.5736, 828.5737, 828.5742, 828.5743, 828.7202, 828.7207,
829.5604, 829.5856, 829.5859, 829.7239, 829.7242, 829.7244,
829.7246, 830.5894, 830.7352, 830.7355, 830.7362, 830.7363,
831.5759, 831.5760, 831.5995, 831.5997, 831.5998, 831.6000,
831.6001, 831.7387, 831.7408, 832.5791, 832.5792, 832.6024,
832.6026, 832.6027, 832.6036, 832.6037, 832.7492, 832.7495,
832.7521, 832.7522, 832.7523, 833.5931, 833.5932, 833.7551,
833.7558, 833.7570, 833.7571, 835.6998, 835.7001, 835.7006,
836.7076, 837.5027, 837.5881, 837.7180, 838.7226, 838.7232,
839.6031, 840.6063, 842.7387, 847.5316, 847.5953, 847.5954,
851.5698, 852.5738, 852.7250, 853.5854, 853.5855, 853.5862,
853.7296, 854.5902, 855.6015, 855.6016, 855.6023, 855.6025,
855.7417, 856.6061, 856.6063, 856.6720, 856.7481, 857.6186,
857.7531, 858.6211, 858.6212, 858.6860, 858.6861, 858.7632,
858.7663, 859.7694, 859.7695, 859.7696, 859.7706, 860.7752,
860.7753, 860.7756, 861.7801, 861.7806, 863.6876, 863.7336,
864.7380, 864.7596, 865.7486, 865.7487, 866.7532, 866.7533,
867.7579, 867.7581, 870.7307, 871.5527, 871.5528, 871.5529,
871.5934, 871.5935, 874.7645, 878.7400, 879.5999, 879.7454,
880.6035, 880.7516, 880.7566, 881.7558, 881.7616, 882.7660,
882.7723, 883.7705, 883.7765, 884.7801, 884.7877, 885.7854,
885.7922, 886.5582, 886.5584, 886.7916, 886.7917, 886.8012,
887.7352, 887.7999, 887.8001, 887.8005, 888.7394, 889.7492,
890.7535, 894.7810, 908.7832, 909.7882, 910.7968, 911.8032,
913.7513, 914.7583, 915.5191, 915.5192, 915.7681, 916.7743,
921.8142, 921.8145, 921.8153, 928.3505, 931.7695, 942.7879,
1018.9420, 1019.3838, 1019.3840, 1085.3294, 1098.9739, 1098.9740,
1225.0920, 1226.0968, 1226.0970, 1227.1070, 1228.1110, 1228.1111
and combinations thereof.
27. The method of claim 26, wherein the accurate mass intensity
data is obtained using a Fourier transform ion cyclotron resonance,
time of flight, orbitrap, quadrupole or triple quadrupole mass
spectrometer.
28. The method of claim 26, wherein the blood sample is a blood
serum sample.
29. The method of claim 26, wherein the accurate mass intensities
represent ionized metabolites.
30. The method of claim 26, wherein a liquid/liquid extraction is
performed on the blood sample whereby non-polar metabolites are
dissolved in an organic solvent and polar metabolites are dissolved
in an aqueous solvent.
31. The method of claim 30, wherein the accurate mass intensities
are obtained from the ionization of the extracted samples using an
ionization method selected from the group consisting of: positive
electrospray ionization, negative electrospray ionization, positive
atmospheric pressure chemical ionization, negative atmospheric
pressure chemical ionization, and combinations thereof.
32. The method of claim 26, wherein the accurate mass intensity
data is obtained using a Fourier transform ion cyclotron resonance
mass spectrometer.
33. The method of claim 26, wherein said one or more than one
reference sample is: a) one or more than one reference sample
obtained from a non-demented control individual; b) one or more
than one reference sample obtained from a demented subject; c) one
or more than one reference sample obtained from a subject
pathologically diagnosed as having Alzheimer's Disease; d) one or
more than one reference sample obtained from a subject clinically
diagnosed as having Alzheimer's Disease; e) one or more than one
reference sample obtained from a patient with cognitive impairment
as measured by Alzheimer's Disease Assessment Scale-cognitive
subset (ADAS-cog); f) one or more than one reference sample
obtained from a patient with cognitive impairment as measured by
Folstein's Mini-Mental State Exam (MMSE); or g) any combination of
a) to above.
34. The method of claim 26, wherein the method is for evaluating
the efficacy of a therapy for treating: (a) Alzheimer's disease
(AD); (b) AD pathology; (c) dementia; (d) cognitive impairment; or
(e) a combination of (a) to (d) above.
35. The method of claim 26, further comprising analyzing a blood
sample from said patient by mass spectrometry to obtain accurate
mass intensity data for one or more than one internal control
metabolite; and calculating a ratio for each of the accurate mass
intensities obtained in step (a) to the accurate mass intensities
obtained for the one or more than one internal control metabolite;
wherein the comparing step (b) comprises comparing each ratio to
one or more corresponding ratios obtained for one or more than one
reference sample.
36. The method of claim 26, wherein an increase in accurate mass
intensity is identified in the comparing step (b).
37. The method of claim 26, wherein the hydrogen and electron
adjusted accurate mass, or neutral accurate mass, is selected from
the group consisting of: 699.5198, 723.5195, 723.5197, 751.5555,
541.3432, 569.3687, 803.568, 886.5582, 565.3394, 569.369, 801.555,
857.6186, 207.0822, 275.8712, 371.7311, 373.728, 432.1532,
485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493, 742.2972,
701.53591, 699.52026, 723.52026, 747.52026, 729.56721, 727.55156,
779.58286, 775.55156, and combinations thereof.
38. A method for evaluating the efficacy of a therapy for treating
Alzheimer's disease, dementia, or cognitive impairment in a
patient, comprising the steps of: a) analyzing a blood sample from
said patient to obtain quantifying data for one or more than one
metabolite marker; b) comparing the quantifying data for said one
or more than one metabolite marker to corresponding data obtained
for one or more than one reference sample to identify an increase
or decrease in the level of said one or more than one metabolite
marker in said blood sample; and c) using said increase or decrease
in the level of said one or more than one metabolite marker in said
blood sample to determine whether the therapy is improving the
biochemical state of the patient, wherein the one or more than one
metabolite marker comprises one or more molecule selected from the
group consisting of: phosphatidylethanolamine (PtdEt),
plasmenylethanolamine, plasmanylethanolamine, and combinations
thereof, wherein the PtdEt is selected from the group consisting
of: PtdEt 16:0/18:0, PtdEt 16:0/18:1, PtdEt 18:0/18:0, PtdEt
18:0/18:1, and combinations thereof.
39. The method of claim 38, wherein the plasmanylethanolamine is
selected from the group consisting of: plasmanyl 16:0/18:1,
plasmanyl 16:0/18:2, plasmanyl 16:0/20:4, plasmanyl 16:0/22:4,
plasmanyl 16:0/22:6, plasmanyl 18:0/18:1, plasmanyl 18:0/18:2,
plasmanyl 18:0/20:4, plasmanyl 18:0/22:4, plasmanyl 18:0/22:6, and
combinations thereof; and, the plasmenylethanolamine is selected
from the group consisting of: plasmenyl 16:0/18:1, plasmenyl
16:0/18:2, plasmenyl 16:0/20:4, plasmenyl 16:0/22:4, plasmenyl
16:0/22:6, plasmenyl 18:0/18:1, plasmenyl 18:0/18:2, plasmenyl
18:0/20:4, plasmenyl 18:0/22:4, plasmenyl 18:0/22:6, and
combinations thereof.
40. The method of claim 38, wherein the quantifying data is
obtained using a Fourier transform ion cyclotron resonance, time of
flight, orbitrap, quadrupole or triple quadrupole mass
spectrometer.
41. The method of claim 40, wherein the mass spectrometer is
equipped with a chromatographic system.
42. The method of claim 38, wherein the blood sample is a blood
serum sample.
43. The method of claim 38, wherein a liquid/liquid extraction is
performed on the blood sample whereby non-polar metabolites are
dissolved in an organic solvent and polar metabolites are dissolved
in an aqueous solvent.
44. The method of claim 43, wherein the extracted samples are
analyzed by positive or negative electrospray ionization or
positive or negative atmospheric pressure chemical ionization.
45. The method of claim 43, wherein the extracted samples are
analyzed by MS/MS transition.
46. The method of claim 43, wherein the extracted samples are
analyzed by extracted ion current (EIC) chromatography and MS/MS
transition.
47. The method of claim 38, wherein said one or more than one
reference sample is a) one or more than one reference sample
obtained from a non-demented control individual; b) one or more
than one reference sample obtained from a demented subject; c) one
or more than one reference sample obtained from a subject
pathologically diagnosed as having Alzheimer's Disease; d) one or
more than one reference sample obtained from a subject clinically
diagnosed as having Alzheimer's Disease; e) one or more than one
reference sample obtained from a patient with cognitive impairment
as measured by Alzheimer's Disease Assessment Scale-cognitive
subset (ADAS-cog); f) one or more than one reference sample
obtained from a patient with cognitive impairment as measured by
Folstein's Mini-Mental State Exam (MMSE); or g) any combination of
a) to f) above.
48. The method according to claim 38, further comprising: analyzing
a blood sample from said patient to obtain quantifying data for one
or more than one internal control metabolite; and obtaining a ratio
for each of the levels of said one or more than one metabolite
marker to the level obtained for the one or more than one internal
control metabolite; wherein the comparing step (b) comprises
comparing each ratio to one or more corresponding ratios obtained
for the one or more than one reference sample.
49. The method of claim 38, wherein an increase in the level of
said one or more than one metabolite marker in said blood sample is
identified in the comparing step (b).
50. The method of claim 39, wherein the extracted samples are
analyzed by MS/MS transition, and the MS/MS transitions for the
phosphatidylethanolamines are 718.0/255.0, 716.0/255.0, 746.0/283.0
and 744.0/283.0, respectively; the MS/MS transitions for the
plasmanylethanolamines are 702.0/281.0, 700.0/279.0, 724.0/303.0,
752.0/331.0, 748.0/327.0, 730.0/281.0, 728.0/279.0, 752.0/303.0,
780.0/331.0 and 776.0/327.0, respectively; and the MS/MS
transitions for the plasmenylethanolamines are 700.0/281.0,
698.0/279.0, 722.0/303.0, 750.0/331.0, 746.0/327.0, 728.0/281.0,
726.0/279.0, 750.6/303.2, 778.0/331.0 and 774.0/327.0,
respectively.
51. The method of claim 26, wherein at least one of the one or more
than one reference sample defined in step (b) is a pre-therapy
baseline blood sample obtained from the patient.
52. The method of claim 1, wherein substantially equivalent refers
to .+-.5 ppm of the hydrogen and electron adjusted accurate mass,
or neutral accurate mass.
53. The method of claim 1, wherein substantially equivalent refers
to .+-.1 ppm of the hydrogen and electron adjusted accurate mass,
or neutral accurate mass.
54. The method of claim 26, wherein substantially equivalent refers
to .+-.5 ppm of the hydrogen and electron adjusted accurate mass,
or neutral accurate mass.
55. The method of claim 26, wherein substantially equivalent refers
.+-.1 ppm of the hydrogen and electron adjusted accurate mass, or
neutral accurate mass.
56. The method of claim 23, wherein the one or more than one
internal control metabolite is a phosphatidylethanolamine
(PtdEt).
57. The method of claim 56, wherein the phosphatidylethanolamine is
selected from the group consisting of PtdEt 16:0/18:0, PtdEt
16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, and combinations
thereof.
58. The method of claim 48, wherein the one or more than one
internal control metabolite is a phosphatidylethanolamine
(PtdEt).
59. The method of claim 58, wherein the phosphatidylethanolamine is
selected from the group consisting of PtdEt 16:0/18:0, PtdEt
16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, and combinations
thereof.
Description
[0001] This application is a continuation of U.S. patent
application Ser. No. 12/280,920, filed Feb. 28, 2007, a national
stage application under 35 U.S.C. .sctn.371 from PCT Application
No. PCT/CA2007/000313, filed Feb. 28, 2007, which claims the
priority benefit of U.S. Provisional Application Nos. 60/888,883,
filed Feb. 8, 2007, 60/804,779, filed Jun. 14, 2006, and
60/777,290, filed Feb. 28, 2006, each of which is hereby
incorporated by reference in its entirety.
FIELD OF INVENTION
[0002] The present invention relates to small molecules or
metabolites that are found to have significantly different
abundances between clinically diagnosed dementia or other
neurological disorders, and normal patients. The present invention
also relates to methods for diagnosing dementia and other
neurological disorders.
BACKGROUND OF THE INVENTION
[0003] The most severe consequence of the aging brain is dementia,
which is defined in the Diagnostic and Statistical Manual of Mental
Disorders, 4th edition (DSM-IV) as [0004] "the development of
multiple cognitive deficits that included memory impairment and at
least one of the following cognitive disturbances: aphasia,
apraxia, agnosia, or a disturbance in executive functions. The
cognitive impairment must be sufficiently severe to cause
impairment in occupational or social functioning and must represent
a decline from a previously higher level of functioning." [1]
[0005] The number of elderly people is increasing rapidly within
our society and as a consequence, dementia is growing into a major
health problem. In 1991, the Canadian Study of Health and Aging had
estimated 25% of the population over the age of 65 had a form of
dementia. The study also estimated the number of people living with
dementia will double and triple in Canada by 2011 and 2031,
respectively [2].
[0006] The clinical manifestation of dementia can result from
neurodegeneration (e.g. Alzheimer's Disease [AD], dementia with
Lewy bodies [DLB] and frontotemporal lobe dementia [FTLD]),
vascular (e.g. multi-infarct dementia) or anoxic event (e.g.
cardiac arrest), trauma to the brain (e.g. dementia pugilistica
[boxer's dementia]), or exposure to an infectious (e.g.
Creutzfeldt-Jakob Disease) or toxic agent (e.g. alcohol-induced
dementia) [3].
[0007] AD is the most common cause of dementia, followed by
vascular dementia (VaD), DLB and FTLD [4]. The differential
diagnosis of the types of dementia is not straightforward, and is
typically based on exclusion of other disorders [5]. For example,
blood chemistry values are measured to determine if Vitamin B12
deficiency, anemia, infection, venereal disease or thyroid disorder
may be possible reasons for the dementia symptoms. Various
neuroimaging techniques may be employed, such as magnetic resonance
imaging or computerized tomography scans to determine if the
symptoms may be due to the presence of a tumor, infection or
vascular event [4].
[0008] If the dementia symptoms can not be explained by another
disorder, a diagnosis of AD, DLB or FTLD is made exclusively based
on the clinical symptoms (e.g. frequency of falls, rapid onset,
presence of visual or auditory hallucinations, etc). It is not
until a histopathological evaluation of the brain during autopsy is
performed that a definitive diagnosis can be obtained [5-7]. A
prospective study on the prevalence of AD in people over the age of
85 indicated that more than half of the individuals with
neuropathological criteria for AD were either non-demented or were
incorrectly diagnosed with VaD. As well, 35% of those individuals
diagnosed with AD based on clinical features were incorrectly
diagnosed as the neuropathological evaluation did not support that
diagnosis [8]. The degree of misdiagnosis is understandable since
the clinical symptoms of the various dementias often overlap and is
dependent upon whether the pertinent information is made known to
the clinician.
[0009] The different types of dementias are based on specific
neuropathological features. A definitive diagnosis of AD relies on
the deposition of two types of neuronal protein: tau in the form of
intraneuronal neurofibrillary tangles (NFTs) and the accumulation
of extracellular .beta.-amyloid to form senile plaques (SPs). Tau
is important for the formation of microtubules in the neuronal axon
by binding and promoting polymerization of tubules. In AD, tau
becomes hyperphosphorylated thereby disrupting its main function.
The tau accumulates and forms tangles within the axon. The neuron
can no longer function and dies. Tau protein is released into the
extracellular space where it can be detected in the cerebrospinal
fluid (CSF) [9]. The formation of SPs, however, is due to the
accumulation of a 40 and 42 residue protein .beta.-amyloid from
amyloid precursor protein (APP) [10]. The formation and secretion
of .beta.-amyloid is closely regulated by homeostasis, but
something occurs in AD that disrupts homeostasis resulting in the
accumulation of the protein within the brain and disrupting the
neurons within its vicinity [11-12]. The increased amount of tau
and the absence of .beta.-amyloid in CSF have been proposed as
possible diagnostic markers for AD, but results have not been
consistent. The problem may be due to the presence of NFTs and SPs
that increase in number during normal aging [13]. In order for the
NFTs and SPs to be diagnostic of AD, they must be localized
together in specific areas of the brain (neocortex and limbic
region) [12]. SPs without NFTs are present in the same area in
individuals with mild cognitive impairment (MCI) and in 27% of
non-demented individuals greater then 75 years old [13].
[0010] A diagnosis of DLB is based on the presence of protein
deposits called alpha-synuclein, which is referred to as Lewy
Bodies, within brainstem and cortical neurons [6]. The cognitive
deficit corresponds to the amount of Lewy Bodies within the
brain.
[0011] FTLD is not characterized by a specific neuropathological
feature. Typically, areas of the frontal/temporal cortices have
neuronal loss, spongiform changes (microvacuolation) and severe
astrocytic gliosis. The clinical symptoms in FTLD are dependent
upon where the pathology is found rather than the type of pathology
[7].
[0012] Currently, various neuropsychological tests are used to help
diagnose dementia. For example, the Alzheimer's Disease Assessment
Scale (ADAS)-cognitive subset is used to test the language ability
(speech and comprehension), memory, ability to copy geometric
figures and orientation to current time and place. The Folstein's
Mini-Mental State Exam (MMSE), which also measures cognitive
impairment, is an extensively validated test of orientation, short
and long-term memory, praxis, language and comprehension. While
these tests may indicate the level of cognitive impairment in an
individual, they give no indication of whether the dementia may be
caused by AD or by non-AD dementias.
[0013] It is commonly accepted that by the time any symptom is
evident in any of the dementias described, irreversible neuronal
loss has occurred [14]. MCI is characterized by a prominent
impairment in memory with normal cognitive functions [15]. MCI is
considered a transitional stage between normal aging and several
types of dementia since a large proportion of individuals with MCI
are later diagnosed with AD, DLB, or FTLD and all individuals with
fully developed dementia first exhibit mild dementia symptoms
similar to MCI [16].
[0014] There is a need to objectively differentiate the types of
dementia from one another. Preferably, such a method would be
specific, accurate, and efficient. Clearly, there is a pressing
need for differential diagnosis of dementia prior to autopsy.
[0015] A biomarker that could detect neuropathological changes
prior to clinical symptoms would be of enormous value. A consensus
was reached in 1999 [17] as to what would be expected in a
biomarker in AD: [0016] 1. Detect a fundamental feature of
neuropathology [0017] 2. Diagnostic sensitivity of >80% for
detecting AD [0018] 3. Specificity of >80% for distinguishing
other dementias [0019] 4. Reliable [0020] 5. Reproducible [0021] 6.
Noninvasive [0022] 7. Simple to perform [0023] 8. Inexpensive
[0024] The identification of AD-specific biomarkers in human serum
would be extremely useful since it would be noninvasive and could
be used to detect the presence of AD pathology prior to the
manifestation of clinical symptoms and differentiate those patients
who may have a different form of dementia but similar clinical
symptoms.
SUMMARY OF THE INVENTION
[0025] The present invention relates to small molecules or
metabolites that are found to have significantly different
abundances between clinically diagnosed dementia or other
neurological disorders, and normal patients. The present invention
also relates to methods for diagnosing dementia and other
neurological disorders.
[0026] The present invention provides a method of identifying one
or more than one metabolite marker for differentially diagnosing AD
dementia, non-AD dementia, cognitive impairment, or a combination
thereof, comprising the steps of: [0027] introducing one or more
than one sample from one or more than one patient with clinically
diagnosed AD dementia, clinically diagnosed non-AD dementia,
significant cognitive impairment, or any combination thereof, said
sample containing a plurality of metabolites into a high resolution
mass spectrometer obtaining quantifying data for the metabolites;
[0028] creating a database of said quantifying data; [0029]
comparing the identifying and quantifying data from the sample with
corresponding data from a sample from a reference sample; [0030]
identifying one or more than one metabolite marker that differs
between same sample and said reference sample,
[0031] wherein the metabolites metabolite markers are selected from
the metabolites listed in Tables 1-7, 10-13, and 18, or any
combination thereof. The method may further comprising selecting a
minimal number of metabolite markers needed for optimal diagnosis.
In a non-limiting example, the high resolution mass spectrometer is
a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer
(FTMS).
[0032] The present invention also provides novel compounds selected
from the group consisting of the metabolites listed in Tables 7-13.
The 15. The metabolite may be selected from the group consisting of
phosphatidylcholine-related compounds, ethanolamine plasmalogens,
endogenous fatty acids, essential fatty acids, lipid oxidation
byproducts, metabolite derivatives of said metabolite classes, and
any metabolite that may contribute in any way to the
anabolic/catabolic metabolism of said metabolite classes.
[0033] In one embodiment of the present invention, the compounds
may be selected from the group consisting of metabolites with
accurate masses measured in Daltons of, or substantially equivalent
to, 541.3432, 569.3687, 699.5198, 723.5195, 723.5197, 751.5555,
803.568, 886.5582, 565.3394, 569.369, 801.555, and 857.6186. The
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, a) 541.3432, b) 569.3687, c) 699.5198,
d) 723.5195, e) 751.5555, and f) 803.568 may also be characterized
by [0034] a) an extracted ion chromatogram (EIC) as shown in FIG.
4A, and an MS/MS spectrum as shown in FIG. 6; [0035] b) an EIC as
shown in FIG. 4B, and an MS/MS spectrum as shown in FIG. 7; [0036]
c) an EIC as shown in FIG. 4C, and an MS/MS spectrum as shown in
FIG. 8; [0037] d) an EIC as shown in FIG. 4D, and an MS/MS spectrum
as shown in FIG. 9; [0038] e) an EIC as shown in FIG. 4E, and an
MS/MS spectrum as shown in FIG. 10; and [0039] f) an EIC as shown
in FIG. 4F, and an MS/MS spectrum as shown in FIG. 11,
respectively.
[0040] The compounds as described above may also be further
characterized by molecular formula a) C.sub.25H.sub.51NO.sub.9P, b)
C.sub.27H.sub.55NO.sub.9P, c) C.sub.39H.sub.74NO.sub.7P, d)
C.sub.41H.sub.74NO.sub.7P, e) C.sub.43H.sub.78NO.sub.7P, and f)
C.sub.43H.sub.81NO.sub.10P, respectively; and/or by the structures
shown in a) FIG. 12; b) FIG. 13; c) FIG. 17; d) FIG. 18; e) FIG.
19; and f) FIG. 14, respectively.
[0041] The compounds may also be selected from the group consisting
of metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, a) 567.3547, b) 565.3394, c) 805.5832,
d) 827.57, e) 829.5856, f) 831.5997, and g) 853.5854. These
compounds may be further characterized by molecular formula a)
C.sub.27H.sub.55NO.sub.9P, b) C.sub.27H.sub.55NO.sub.9P, c)
C.sub.43H.sub.83NO.sub.10Y, C.sub.45H.sub.81NO.sub.10Y, e)
C.sub.45H.sub.83NO.sub.10P, f) C.sub.45H.sub.85NO.sub.10P, and g)
C.sub.47H.sub.83NO.sub.10P, respectively; and/or by the structure
shown in a) FIG. 15A; b) FIG. 15B; c) FIG. 15C; d) FIG. 15D; e)
FIG. 15E; f) FIG. 15F; and g) FIG. 15G, respectively.
[0042] The compounds may further be selected from the group
consisting of metabolites M05 to M24 with accurate masses of, or
substantially equivalent to those listed in Table 18. Of these
compounds, the metabolites with accurate masses measured in Daltons
of, or substantially equivalent to, a) 701.53591, b) 699.52026, c)
723.52026, d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286,
and h) 775.55156 may be further characterized by a MS/MS spectrum
as shown in a) FIG. 21; b) FIG. 22; c) FIG. 23; d) FIG. 24; e) FIG.
25; f) FIG. 26; g) FIG. 27; and h) FIG. 28, respectively.
[0043] The compounds as described above may also be further
characterized by molecular formula a) C.sub.39H.sub.76NO.sub.7P, b)
C.sub.39H.sub.74NO.sub.7P, c) C.sub.41H.sub.74NO.sub.7P, d)
C.sub.43H.sub.74NO.sub.7P, e) C.sub.41H.sub.80NO.sub.7P, f)
C.sub.41H.sub.78NO.sub.7P, C.sub.45H.sub.82NO.sub.7P, and h)
C.sub.45H.sub.78NO.sub.7P, respectively and/or by the structure
##STR00001##
respectively.
[0044] The novel compounds may also be selected from the group
consisting of the metabolites listed in Table 30. Of these
compounds, the metabolites with accurate masses measured in Daltons
of, or substantially equivalent to, 207.0822, 275.8712, 371.7311,
373.728, 432.1532, 485.5603, 487.6482, 562.46, 622.2539, 640.2637,
730.6493, and 742.2972 are of particular interest.
[0045] One or more than one of the compounds of the present
invention may be used for the differential diagnosis of
dementia.
[0046] In another embodiment, the present invention provides a
method for differentially diagnosing dementia or the risk of
dementia in a patient, the method comprising the steps of: [0047]
a) obtaining a sample from said patient; [0048] b) analyzing said
sample to obtain quantifying data for one or more than one
metabolite marker; [0049] c) comparing the quantifying data for
said one or more than one metabolite marker to corresponding data
obtained from one or more than one reference sample; and [0050] d)
using said comparison to differentially diagnose dementia or the
risk of dementia.
[0051] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0052] In the method as just described, the one or more than one
reference sample is a first reference sample obtained from a
non-demented control individual. The one or more than one reference
sample may also comprise a second reference sample obtained from a
patient with clinically diagnosed AD-dementia; a third reference
sample obtained from a patient with clinically diagnosed non-AD
dementia; and/or a fourth reference sample obtained from a patient
suffering from significant cognitive impairment.
[0053] In one alternative of the method described above, the sample
and the reference sample are serum samples, and the one or more
than one metabolite marker is selected from the metabolites listed
in Tables 1 to 7, or a combination thereof. These metabolite
markers may be selected from the group consisting of
phosphatidylcholine-related compounds, ethanolamine plasmalogens,
endogenous fatty acids, essential fatty acids, lipid oxidation
byproducts, metabolite derivatives of said metabolite classes, and
any metabolite that may contribute in any way to the
anabolic/catabolic metabolism of said metabolite classes.
[0054] The one or more than one metabolite marker needed for
optimal diagnosis may comprise metabolites with accurate masses
measured in Daltons of, or substantially equivalent to, 541.3432,
569.3687, 699.5198, 723.5195, 723.5197, 751.5555, 803.568,
886.5582, and any combination thereof. Of these, the metabolite of
accurate masses 699.5198, 723.5195, 723.5197, and 751.555 are
ethanolamine plasmalogens and are specifically decreased in
patients with AD dementia; and the metabolite markers of accurate
masses 541.3432, 569.3687, 803.568, and 886.5582 are
phosphatidylchoine metabolites, are decreased in patients with
cognitive impairment on ADAS-cog, and severity of cognitive
impairment correlates to the degree of decrease.
[0055] The one or more than one metabolite marker may be the
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, a) 541.3432, b) 569.3687, c) 699.5198,
d) 723.5195, e) 751.5555, and f) 803.568. These metabolites may be
further characterized by [0056] a) an extracted ion chromatogram
(EIC) as shown in FIG. 4A, and an MS/MS spectrum as shown in FIG.
6; [0057] b) an EIC as shown in FIG. 4B, and an MS/MS spectrum as
shown in FIG. 7; [0058] c) an EIC as shown in FIG. 4C, and an MS/MS
spectrum as shown in FIG. 8; [0059] d) an EIC as shown in FIG. 4D,
and an MS/MS spectrum as shown in FIG. 9; [0060] e) an EIC as shown
in FIG. 4E, and an MS/MS spectrum as shown in FIG. 10; and [0061]
f) an EIC as shown in FIG. 4F, and an MS/MS spectrum as shown in
FIG. 11,
[0062] respectively. The metabolite may also be further
characterized by molecular formula a) C.sub.25H.sub.51NO.sub.9P, b)
C.sub.27H.sub.55NO.sub.9P, c) C.sub.39H.sub.74NO.sub.7P, d)
C.sub.41H.sub.74NO.sub.7P, e) C.sub.43H.sub.78NO.sub.7P, and f)
C.sub.43H.sub.81NO.sub.10P, respectively; and/or by the structure
shown in a) FIG. 12; b) FIG. 13; c) FIG. 17; d) FIG. 18; e) FIG.
19; and f) FIG. 14, respectively.
[0063] In another alternative of the method described above, the
sample and the reference sample may be cerebrospinal fluid (CSF)
samples, and the one or more than one metabolite marker is selected
from the metabolites listed in Table 13, or a combination thereof.
Of particular interest are metabolite markers needed for optimal
diagnosis, which may comprise metabolites with accurate masses
measured in Daltons of, or substantially equivalent to, 207.0822,
275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482, 562.46,
622.2539, 640.2637, 730.6493, 742.2972, and any combination
thereof. Of these, metabolite markers 207.0822, 432.1532, 562.46,
622.2539, 640.2637, 730.6493, and 742.2972 are increased in
patients with AD dementia; and metabolite markers 275.8712,
371.7311, 373.728, 485.5603, and 487.6482 are decreased in patients
with AD dementia.
[0064] In yet another alternative of the method described above,
the sample and the reference sample are serum samples, and the one
or more than one metabolite marker may be selected from metabolites
M05 to M24 with accurate masses of, or substantially equivalent to
those listed in Table 18. Of these, the one or more than one
metabolite marker of particular interest may comprise metabolites
with accurate masses measured in Daltons of, or substantially
equivalent to, a) 701.53591, b) 699.52026, c) 723.52026, d)
747.52026, e) 729.56721, f) 727.55156, g) 779.58286, and h)
775.55156, and wherein a decrease in the level of a) to h)
indicates AD dementia with a severe cognitive impairment.
[0065] The metabolites listed above may be further characterized by
a MS/MS spectrum as shown in a) FIG. 21, b) FIG. 22, c) FIG. 23, d)
FIG. 24, e) FIG. 25, f) FIG. 26, g) FIG. 27, and h) FIG. 28,
respectively. The metabolites may also be further characterized by
molecular formula a) C.sub.39H.sub.76NO.sub.7P, b)
C.sub.39H.sub.74NO.sub.7P, c) C.sub.41H.sub.74NO.sub.7P, d)
C.sub.43H.sub.74NO.sub.7P, e) C.sub.41H.sub.80NO.sub.7P, f)
C.sub.41H.sub.78NO.sub.7P, g) C.sub.45H.sub.82NO.sub.7P, and h)
C.sub.45H.sub.78NO.sub.7P, respectively; and/or by the
structure
##STR00002##
respectively.
[0066] In yet another aspect of the present invention, there is
provided a method for assessing dementia or the risk of dementia in
a patient, the method comprising the steps of: [0067] a) obtaining
a serum sample from said patient; [0068] b) analyzing said sample
to obtain quantifying data for one or more than one metabolite
marker; [0069] c) comparing the quantifying data for said one or
more than one metabolite marker to corresponding data obtained from
one or more than one reference sample; and [0070] d) using said
comparison to assess dementia or the risk of dementia.
[0071] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0072] In the method as just described, the one or more than one
reference sample may be a first reference sample obtained from a
non-demented control individual. The one or more than one reference
sample may also further comprise a second reference sample obtained
from a patient with cognitive impairment as measured by ADAS-cog,
and/or a third reference sample obtained from a patient with
cognitive impairment as measured by MMSE.
[0073] The one or more than one metabolite marker in the method
described above may be selected from the metabolites listed in
Tables 10-12, or a combination thereof. Of particular interest are
the one or more than one metabolite markers is selected from the
group consisting of metabolites with accurate masses measured in
Daltons of, or substantially equivalent to 541.3432, 569.3687,
699.5198, 723.5195, 723.5197, 751.5555, 803.568, 886.5582,
565.3394, 569.369, 801.555, 857.6186, and any combination thereof.
Of these, a decrease in the patient sample in metabolite markers
699.5198, 723.5195, 723.5197, and 751.555 indicates AD pathology; a
decrease in the patient sample in metabolite markers 541.3432,
569.3687, 803.568, and 886.5582 indicates cognitive impairment on
ADAS-cog; and a decrease in the patient sample in metabolite
markers 565.3394, 569.369, 801.555, and 857.6186 indicates
cognitive impairment on MMSE.
[0074] In yet another embodiment of the present invention, a method
is provided for differentially diagnosing dementia or the risk of
dementia in a patient, the method comprising the steps of: [0075]
a) obtaining a sample from said patient; [0076] b) analyzing said
sample to obtain quantifying data for one or more than one
metabolite marker; [0077] c) obtaining a ratio for each of the one
or more than one metabolite marker to an internal control
metabolite; [0078] d) comparing each ratio of said one or more than
one metabolite marker to the internal control metabolite to
corresponding data obtained from one or more than one reference
sample; and [0079] e) using said comparison to differentially
diagnose dementia or the risk of dementia.
[0080] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0081] In the method as just described, the one or more than one
reference sample may be a first reference sample obtained from a
non-demented control individual. The one or more than one reference
sample may further comprise a second reference sample obtained from
a patient with clinically diagnosed AD-dementia; a third reference
sample obtained from a patient with clinically diagnosed non-AD
dementia; and/or a fourth reference sample obtained from a patient
suffering from significant cognitive impairment.
[0082] In one aspect of the method described above, the sample and
the reference sample are serum samples, and the one or more than
one metabolite marker is selected from metabolites M05 to M24 with
accurate masses of, or substantially equivalent to those listed in
Table 18. Of particular interest is the one or more than one
metabolite marker comprising metabolites with accurate masses
measured in Daltons of, or substantially equivalent to, a)
701.53591, b) 699.52026, c) 723.52026, d) 747.52026, e) 729.56721,
f) 727.55156, g) 779.58286, and h) 775.55156, and the internal
control metabolite comprising the metabolite with accurate mass
measured in Daltons of, or substantially equivalent to, 719.54648.
When these metabolites and internal control metabolite are used, a
decrease in the ratio of metabolite to the internal control
metabolite indicates AD dementia with a severe cognitive
impairment.
[0083] The metabolites described above may be further characterized
by a MS/MS spectrum as shown in a) FIG. 21, b) FIG. 22, c) FIG. 23,
d) FIG. 24, e) FIG. 25, f) FIG. 26, g) FIG. 27, and h) FIG. 28,
respectively. These metabolites may also be further characterized
by molecular formula a) C.sub.39H.sub.76NO.sub.7P, b)
C.sub.39H.sub.74NO.sub.7P, c) C.sub.41H.sub.74NO.sub.7P, d)
C.sub.43H.sub.74NO.sub.7P, e) C.sub.41H.sub.80NO.sub.7P,
C.sub.41H.sub.78NO.sub.7P, g) C.sub.45H.sub.82NO.sub.7P, and h)
C.sub.45H.sub.78NO.sub.7P, respectively, and the internal control
metabolite may be characterized by molecular formula
C.sub.39H.sub.78NO.sub.8P; and/or by the structure
##STR00003##
respectively, and the internal control metabolite may be further
characterized by the structure
##STR00004##
[0084] In yet another embodiment of the present invention, there is
provided a method for evaluating the efficacy of a therapy for
treating dementia in a patient, comprising: [0085] a) obtaining a
sample from said patient; [0086] b) analyzing said sample to obtain
quantifying data for one or more than one metabolite marker; [0087]
c) comparing said quantifying data to corresponding data obtained
from one or more than one reference sample; and [0088] d) using
said comparison to determine whether the therapy is improving the
demented state of the patient.
[0089] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0090] In the method as just described, the one or more than one
reference sample may be a plurality of samples obtained from a
non-demented control individuals; a plurality of samples obtained
from a clinically diagnosed AD patient; one or more than one
pre-therapy baseline sample obtained from the patient; or any
combination thereof.
[0091] In one aspect of the above method, the sample and the
reference sample are serum samples, and the one or more than one
metabolite marker is selected from the metabolites listed in Tables
1 to 7, or a combination thereof. These metabolite marker markers
needed for optimal diagnosis may be selected from the group
consisting of phosphatidylcholine-related compounds, ethanolamine
plasmalogens, endogenous fatty acids, essential fatty acids, lipid
oxidation byproducts, metabolite derivatives of said metabolite
classes, and any metabolite that may contribute in any way to the
anabolic/catabolic metabolism of said metabolite classes. Of
particular interest are the metabolites with accurate masses
measured in Daltons of, or substantially equivalent to, 541.3432,
569.3687, 699.5198, 723.5195, 723.5197, 751.5555, 803.568,
886.5582.
[0092] In another aspect, the sample and the reference sample are
cerebrospinal fluid (CSF) samples, and the one or more than one
metabolite marker is selected from the metabolites listed in Table
13, or a combination thereof. Of particular interest are the
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, 207.0822, 275.8712, 371.7311, 373.728,
432.1532, 485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493,
742.2972.
[0093] In a third aspect, the sample and the reference sample are
serum samples, and the one or more than one metabolite marker may
be selected from metabolites M05 to M24 with accurate masses of, or
substantially equivalent to those listed in Table 18. Of these
metabolites, the metabolites with accurate masses measured in
Daltons of, or substantially equivalent to, 701.53591, 699.52026,
723.52026, 747.52026, 729.56721, 727.55156, 779.58286, and
775.55156 may be of particular interest.
[0094] The present invention also provides a method for evaluating
the efficacy of a therapy for treating dementia in a patient,
comprising: [0095] a) obtaining a sample from said patient; [0096]
b) analyzing said sample to obtain quantifying data for one or more
than one metabolite marker; [0097] c) obtaining a ratio for each of
the one or more than one metabolite marker to an internal control
metabolite; [0098] d) comparing each ratio of said one or more than
one metabolite marker to the internal control metabolite to
corresponding data obtained from one or more than one reference
sample; and [0099] e) using said comparison to determine whether
the therapy is improving the demented state of the patient.
[0100] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0101] In the method as just described, the one or more than one
reference sample may be a plurality of samples obtained from a
non-demented control individuals; a plurality of samples obtained
from a clinically diagnosed AD patient; one or more than one
pre-therapy baseline sample obtained from the patient; or any
combination thereof.
[0102] In the method as described above, the sample and said
reference sample are serum samples, and the one or more than one
metabolite marker may be selected from metabolites M05 to M24 with
accurate masses of, or substantially equivalent to those listed in
Table 18. Of particular interest are the metabolites with accurate
masses measured in Daltons of, or substantially equivalent to,
701.53591, 699.52026, 723.52026, 747.52026, 729.56721, 727.55156,
779.58286, and 775.55156, and the internal control metabolite with
accurate mass measured in Daltons of, or substantially equivalent
to, 719.54648.
[0103] The methods of the present invention, including HTS assays,
can be used for the following, wherein the specific "health-state"
in this application refers to, but is not limited to dementia:
[0104] 1. identifying small-molecule metabolite biomarkers which
can discriminate between multiple health-states using any
biological sample taken from an individual,
[0105] 2. specifically diagnosing a health-state using metabolites
identified in serum, plasma, whole blood, serum, CSF, and/or other
tissue biopsy as described in this application,
[0106] 3. selecting the minimal number of metabolite features
required for optimal diagnostic assay performance statistics using
supervised statistical methods such as those mentioned in this
application,
[0107] 4. identifying structural characteristics of biomarker
metabolites selected from non-targeted metabolomic analysis using
LC-MS/MS, MSn and NMR,
[0108] 5. developing a high-throughput LC-MS/MS method for assaying
selected metabolite levels in serum,
[0109] 6. diagnosing a given health-state, or risk for development
of a health-state by determining the levels of any combination of
metabolite features disclosed from the FTMS analysis patient serum,
using any method including but not limited to mass spectrometry,
NMR, UV detection, ELISA (enzyme-linked immunosorbant assay),
chemical reaction, image analysis, or other.
[0110] The impact of the present invention on the diagnosis of
dementia would be tremendous, as literally everyone could be
screened longitudinally throughout their lifetime to assess risk.
Given that the performance characteristics of the test of the
present invention are representative for the general population,
this test alone may be superior to any other currently available
screening method, as it may have the potential to detect disease
progression prior to the emergence of clinical symptoms.
[0111] This summary of the invention does not necessarily describe
all features of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0112] These and other features of the invention will become more
apparent from the following description in which reference is made
to the appended drawings wherein:
[0113] FIG. 1 shows a mean signal-to-noise +/-SEM of the AD serum 8
biomarker panel for each different clinical group (AD with
significant cognitive impairment, Non-AD dementia, and AD with no
significant cognitive impairment) relative to non-demented
controls.
[0114] FIG. 2 shows a mean signal-to-noise +/-SEM of the AD serum 8
biomarker panel for two clinical groups with a significant
cognitive impairment (AD and Non-AD dementia).
[0115] FIG. 3 shows a mean signal-to-noise +/-SEM of the AD CSF 12
biomarker panel for two clinical groups with a significant
cognitive impairment (AD and Non-AD dementia).
[0116] FIG. 4 shows Q-Star extracted ion chromatograms (EIC) for
the metabolites 541.3432 (A), 569.3687 (B), 699.5198 (C), 723.5195
(D), 751.5555 (E), and 803.568 (F). Top panel, 8 samples from
non-demented subjects, bottom panel, 8 samples from
clinically-diagnosed AD subjects.
[0117] FIG. 5 shows averaged AD biomarker intensities of the 8 AD
and 8 non-demented controls samples from FTMS and Q-Star
Analysis.
[0118] FIG. 6 shows MS/MS spectra for metabolite 541.3432 with CE
voltage -50V.
[0119] FIG. 7 shows MS/MS spectra for metabolite 569.3687 with CE
voltage -50V.
[0120] FIG. 8 shows MS/MS spectra for metabolite 699.5198 with CE
voltage -50V.
[0121] FIG. 9 shows MS/MS spectra for metabolite 723.5195 with CE
voltage -50V.
[0122] FIG. 10 shows MS/MS spectra for metabolite 751.5555 with CE
voltage -50V.
[0123] FIG. 11 shows MS/MS spectra for metabolite 803.568 with CE
voltage -50V.
[0124] FIG. 12 shows structural determination of ADAS-cog serum
biomarker 541.3432.
[0125] FIG. 13 shows structural determination of ADAS-cog serum
biomarker 569.3687.
[0126] FIG. 14 shows structural determination of ADAS-cog serum
biomarker 803.568.
[0127] FIG. 15 shows putative structures of additional serum
biomarkers. A--metabolite with mass of 567.3547; B--metabolite with
mass of 565.3394; C--metabolite with mass of 805.5832;
D--metabolite with mass of 827.57; E--metabolite with mass of
829.5856; F--metabolite with mass of 531.5997; and G--metabolite
with mass of 853.5854.
[0128] FIG. 16 shows the fragments obtained for the MS/MS analysis
of the 751.5555 metabolite, along with its proposed structure.
[0129] FIG. 17 shows the fragments obtained for the MS/MS analysis
of the 699.5198 metabolite, along with its proposed structure.
[0130] FIG. 18 shows the fragments obtained for the MS/MS analysis
of the 723.5195 metabolite, along with its proposed structure.
[0131] FIG. 19 shows the LC-MS and MS/MS analysis of the 751.5555
metabolite (18:0/20:4 EtnPls). Panel A1 is an extracted ion
chromatogram (EIC) of parent ion 750 (M--H--) of a pure standard;
panel A2 is MS/MS spectra of parent ion M/Z 750 @ retention time
4.8-5.0 minutes. Panel B1 is the EIC of parent ion 750 from a
cognitively normal subject; panel B2 is the MS/MS spectra of parent
ion M/Z 750 @ 4.8-5.0 min. Panel C1 is the EIC of parent ion 750
from an AD subject; and panel C2 is the MS/MS spectra of parent ion
M/Z 750@ 4.8-5.0 min.
[0132] FIG. 20 shows the general structure of ethanolamine
phospholipids, as well as the naming convention used herein.
[0133] FIG. 21 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 16:0/18:1 (M15) in human
serum.
[0134] FIG. 22 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 16:0/18:2 (M16) in human
serum.
[0135] FIG. 23 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 16:0/20:4 (M17) in human
serum.
[0136] FIG. 24 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 16:0/22:6 (M19) in human
serum.
[0137] FIG. 25 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 18:0/18:1 (M20) in human
serum.
[0138] FIG. 26 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 18:0/18:2 (M21) in human
serum.
[0139] FIG. 27 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 18:0/20:4 (M23) in human
serum.
[0140] FIG. 28 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 18:0/22:6 (M24) in human
serum.
[0141] FIG. 29 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 18:1/18:2 and Plasmanyl
16:0/20:4 (M07) in human serum.
[0142] FIG. 30 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of EtnPls 20:0/20:4 and EtnPls
18:0/22:4 (M23) in human serum.
[0143] FIG. 31 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panel) of Plasmanyl 18:0/20:4 (M12) and
Plasmanyl 16:0/22:4 (M08) in human serum.
[0144] FIG. 32 is an extracted ion chromatogram (upper panel) and
MS/MS spectrum (lower panels) of EtnPls 18:1/20:4, EtnPls
16:0/22:5, Plasmanyl 16:0/22:6 (M09) in human serum.
[0145] FIG. 33 shows a Q-Trap flow injection analysis standard
curve of EtnPls 16:0/22:6 (M19) in healthy human serum.
[0146] FIG. 34 shows the effect of dementia severity and SDAT
pathology on serum EtnPl levels (male and female subjects
combined). (A) Mono and di-unsaturated EtnPls and saturated PtdEt
internal control. (B) Polyunsaturated EtnPls and free DHA (22:6).
EtnPls abbreviations: (fatty acid carbons:double bonds, not
including the vinyl ether double bond) and position on glycerol
backbone (sn-1/sn-2). D16:0/18:0 represents
diacylglycerophosphatidylethanolamine with palmitic acid (16:0) at
sn-1 and stearic acid (18:0) at sn-2; 22:6 represents free DHA.
Values are expressed as mean+SEM (n=19-112).
[0147] FIG. 35 shows serum DHA-EtnPls (Log(2) EtnPls 16:0/22:6
(M19) to PtdEt 16:0/18:0 (M01) ratio) distributions in subjects
with different levels of dementia severity (male and female
subjects combined).
[0148] FIG. 36 is a comparison of theoretical distributions of AD
pathology (A) compiled from references 5-8 and experimentally
determined distributions of serum 22:6-containing EtnPls (Log(2)
EtnPls 16:0/22:6 (M19) to PtdEt 16:0/18:0 (M01) ratio) (B) in
cognitively normal and demented subjects. Arrow indicates positive
diagnosis of AD.
[0149] FIG. 37 is a linear regression analysis of disease severity
(ADAS-cog) and serum 22:6-containing EtnPls (EtnPls 16:0/22:6 (M19)
to PtdEt 16:0/18:0 (M01) ratio) levels in 256 AD subjects.
X=predicted initiation of EtnPls depletion. Values are expressed as
mean.+-.SEM (n=66-112). Clinical progression assumes 7.5 ADAS-cog
points/year.
[0150] FIG. 38 shows serum 22:6-containing EtnPls (EtnPls 16:0/22:6
(M19) to PtdEt 16:0/18:0 (M01) ratio) levels in AD, Cognitive
Normal, and general population subjects. (A) Mean.+-.SEM
(n=68-256). (B) Log(2) distributions.
[0151] FIG. 39 shows the distribution of serum white and gray
matter EtnPl scores in males and females.
DETAILED DESCRIPTION
[0152] The present invention relates to small molecules or
metabolites that are found to have significantly different
abundances between clinically diagnosed dementia or other
neurological disorders, and normal patients. The present invention
also relates to methods for diagnosing dementia and other
neurological disorders.
[0153] The present invention provides novel methods for
discovering, validating, and implementing a metabolite markers for
one or more diseases or particular health-states. In one embodiment
of the present invention, there is provided a method for
identifying specific biomarkers for differentially diagnosing AD
dementia, non-AD dementia, cognitive impairment, or a combination
thereof, comprising the steps of: introducing one or more than one
sample from one or more than one patient with clinically diagnosed
AD dementia, clinically diagnosed non-AD dementia, or significant
cognitive impairment, said sample containing a plurality of
metabolites into a high resolution mass spectrometer (for example,
and without wishing to be limiting, a Fourier Transform Ion
Cyclotron Resonance Mass Spectrometer (FTMS)); obtaining,
identifying and quantifying data for the metabolites; creating a
database of said identifying and quantifying data; comparing the
identifying and quantifying data from the sample with corresponding
data from a sample from a non-demented normal patient; identifying
one or more than one metabolites that differ. The metabolite
markers identified using the method of the present invention may
include the metabolites listed in Tables 1-7, 10-13, and 18. The
method may further comprise selecting the minimal number of
metabolite markers needed for optimal diagnosis.
[0154] In order to determine the biochemical markers of a given
health-state in a particular population, a group of patients
representative of the health state (i.e. a particular disease)
and/or a group of "normal" counterparts are required. Biological
samples taken from the patients in the particular health-state
category can then be compared to the same samples taken from the
normal population as well as to patients in similar health-state
category in the hopes of identifying biochemical differences
between the two groups, by analyzing the biochemicals present in
the samples using FTMS and/or LC-MS.
[0155] The method for the discovery of metabolite markers as
described above may be done using non-targeted metabolomic
strategies or methods. Multiple non-targeted metabolomics
strategies have been described in the scientific literature
including NMR [18], GC-MS [19-21], LC-MS, and FTMS strategies [18,
22-24]. The metabolic profiling strategy employed for the discovery
of differentially expressed metabolites in the present invention
was the non-targeted FTMS strategy by Phenomenome Discoveries [21,
24-27; see also US Published Application No. 2004-0029120 A1,
Canadian Application No. 2,298,181, and WO 0157518]. Non-targeted
analysis involves the measurement of as many molecules in a sample
as possible, without any prior knowledge or selection of components
prior to the analysis. Therefore, the potential for non-targeted
analysis to discover novel metabolite biomarkers is high versus
targeted methods, which detect a predefined list of molecules. The
present invention uses a non-targeted method to identify metabolite
components in serum samples that differ between clinically
diagnosed AD individuals and non AD individuals. The same
technology was used to identify metabolite components that differ
between clinically diagnosed AD individuals with dementia from
clinically diagnosed non-AD individuals with dementia in CSF
samples.
[0156] However, a person skilled in the art would recognize that
other metabolite profiling strategies could be used to discover
some or all of the differentially regulated metabolites disclosed
in the present invention and that the metabolites described herein,
however discovered or measured, represent unique chemical entities
that are independent of the analytical technology that may be used
to detect and measure them.
[0157] The present invention also provides a method for
differentially diagnosing dementia or the risk of dementia in a
patient, the method comprising the steps of: [0158] a) obtaining a
sample from said patient; [0159] b) analyzing said sample to obtain
quantifying data for one or more than one metabolite marker; [0160]
c) comparing the quantifying data for said one or more than one
metabolite marker to corresponding data obtained from one or more
than one reference sample; and [0161] d) using said comparison to
differentially diagnose dementia or the risk of dementia. The step
of analyzing the sample (steb b) may comprise analyzing the sample
using a mass spectrometer (MS). For example, and without wishing to
be limiting, such mass spectrometer could be of the FTMS, orbitrap,
time of flight (TOF) or quadrupole types. Alternatively, the mass
spectrometer could be equipped with an additional pre-detector mass
filter. For example, and without wishing to be limiting such
instruments are commonly referred to as quadrupole-FTMS (Q-FTMS),
quadrupole-TOF (Q-TOF) or triple quadrupole (TQ or QQQ). In
addition, the mass spectrometer could be operated in either the
parent ion detection mode (MS) or in MSn mode, where n>=2. MSn
refers to the situation where the parent ion is fragmented by
collision induced dissociation (CID) or other fragmentation
procedures to create fragment ions, and then one or more than one
of said fragments are detected by the mass spectrometer. Such
fragments can then be further fragmented to create further
fragments. Alternatively, the sample could be introduced into the
mass spectrometer using a liquid or gas chromatographic system or
by direct injection.
[0162] By the term "differential diagnosis" or "differentially
diagnosing", it is meant that various aspects of a disease state
may be distinguished from one another. In particular, the present
invention allows for differential diagnosis a various states of
dementia; for example and without wishing to be limiting, the
present invention may provide differential diagnosis of AD
dementia, non-AD dementia, cognitive impairment, or a combination
thereof.
[0163] The diagnosis of or the exclusion of any types of
neurological disorders is contemplated by the present invention,
using all or a subset of the metabolites disclosed herein. The term
"dementia" is used herein as a broad term indicating both cognitive
impairment as well as pathologies causing cognitive impairment.
Dementia may be caused by a number of neurological disorders. "AD
dementia" as used herein refers to dementia caused by Alzheimer's
disease (AD, which may also be referred to herein as "SDAT"); types
of "non-AD dementia" include, but are not limited to, dementia with
Lewy bodies (DLB), frontotemporal lobe dementia (FTD), vascular
induced dementia (e.g. multi-infarct dementia), anoxic event
induced dementia (e.g. cardiac arrest), trauma to the brain induced
dementia (e.g. dementia pugilistica [boxer's dementia]), dementia
resulting from exposure to an infectious (e.g. Creutzfeldt-Jakob
Disease) or toxic agent (e.g. alcohol-induced dementia), Autism,
Multiple Sclerosis, Parkinson's Disease, Bipolar Disorder,
Ischemia, Huntington's Chorea, Major Depressive Disorder, Closed
Head Injury, Hydrocephalus, Amnesia, Anxiety Disorder, Traumatic
Brain Injury, Obsessive Compulsive Disorder, Schizophrenia, Mental
Retardation, and/or Epilepsy. Of particular interest are AD
dementia, and FTD and DLB non-AD dementias.
[0164] Cognitive impairment can be assessed by any method known in
the art. For example, and without wishing to be limiting, the
Alzheimer's Disease Assessment Scale (ADAS)-cognitive subset may be
used. This neuropsychological test is used to test the language
ability (speech and comprehension), memory, ability to copy
geometric figures and orientation to current time and place. Errors
on the test are recorded resulting in a reverse score impairment
(i.e., the higher the score on ADAS, the greater the cognitive
impairment). A score of 0-15 is considered normal, 16-47 is
considered mild-moderate impairment and a score of 48-70 is
considered moderate-severe impairment [28]. Another
neuropsychological test, Folstein's Mini-Mental State Exam (MMSE),
which measures cognitive impairment, may be used. The MMSE is
widely used and is an extensively validated test of orientation,
short and long-term memory, praxis, language and comprehension. A
person skilled in the art would recognize that additional
neuropsychological assessment that measure aspects of the same
cognitive deficit, such as, but not exclusive to, the Blessed Roth
Dementia Rating Scale, the 7-Minute Screen, Wechsler Memory Scale
(WMS), Halstead-Reitan Battery, Rey Auditory Verbal Learning Test,
California Verbal Learning Test, Buschke Selective Reminding Test,
Boston Naming Test, Clinical Evaluation of Language Functioning,
Peabody Picture Vocabulary Tests, Mattis Dementia Rating Scale,
Memory Assessment Scale, Tests of Memory and Learning, Wide Range
Assessment of Memory and Learning, can also be used.
[0165] In addition, a person skilled in the art would recognize
that any imaging technique that has the potential to show a
cognitive impairment or structural change, such as, but not
exclusive to, structural magnetic resonance imaging (MRI), positron
emission tomography (PET), computerized tomography (CT), functional
magnetic resonance imaging (fMRI), electroencephalography (EEG),
single positron emission tomography (SPECT), event related
potentials, magnetoencephalography, multi-modal imaging, would be
measuring the structural/regional brain areas that are responsible
for that cognitive deficit and AD pathology, and therefore, would
be related to the metabolites disclosed in this invention.
[0166] In accordance with the present invention, any type of
biological sample that originates from anywhere within the body,
for example but not limited to, blood (serum/plasma), CSF, urine,
stool, breath, saliva, or biopsy of any solid tissue including
tumor, adjacent normal, smooth and skeletal muscle, adipose tissue,
liver, skin, hair, brain, kidney, pancreas, lung, colon, stomach,
or other may be used. Of particular interest are samples that are
serum or CSF. While the term "serum" is used herein, those skilled
in the art will recognize that plasma or whole blood or a
sub-fraction of whole blood may also be used. CSF may be obtained
by a lumbar puncture requiring a local anesthetic.
[0167] In a non-limiting example, when a blood sample is drawn from
a patient there are several ways in which the sample can be
processed. The range of processing can be as little as none (i.e.
frozen whole blood) or as complex as the isolation of a particular
cell type. The most common and routine procedures involve the
preparation of either serum or plasma from whole blood. All blood
sample processing methods, including spotting of blood samples onto
solid-phase supports, such as filter paper or other immobile
materials, are also contemplated by the present invention.
[0168] In another non-limiting example, CSF samples may be
collected using a lumbar puncture procedure; a local anesthetic is
applied to the lower back. A needle is then inserted into the
numbed skin between the L4 and L5 vertebrae until it pierces the
subdural space. The CSF may be collected into sterile tubes.
[0169] For example, but not considered to be limiting in any
manner, while obtaining a CSF sample may result in more discomfort
for the patient than taking a blood sample, a CSF assay used after
a positive result on a AD-specific serum test, a differential
diagnosis between AD and non-AD has a higher degree of
confirmation.
[0170] Without wishing to be limiting in any manner, the processed
blood, serum or CSF sample described above may then be further
processed to make it compatible with the methodical analysis
technique to be employed in the detection and measurement of the
metabolites contained within the processed serum or CSF sample. The
types of processing can range from as little as no further
processing to as complex as differential extraction and chemical
derivatization. Extraction methods could include sonication,
soxhlet extraction, microwave assisted extraction (MAE),
supercritical fluid extraction (SFE), accelerated solvent
extraction (ASE), pressurized liquid extraction (PLE), pressurized
hot water extraction (PHWE) and/or surfactant assisted extraction
(PHWE) in common solvents such as methanol, ethanol, mixtures of
alcohols and water, or organic solvents such as ethyl acetate or
hexane. A method of particular interest for extracting metabolites
for FTMS non-targeted analysis is to perform a liquid/liquid
extraction whereby non-polar metabolites dissolve in an organic
solvent and polar metabolites dissolve in an aqueous solvent.
[0171] The extracted samples may be analyzed using any suitable
method know in the art. For example, and without wishing to be
limiting in any manner, extracts of biological samples are amenable
to analysis on essentially any mass spectrometry platform, either
by direct injection or following chromatographic separation.
Typical mass spectrometers are comprised of a source which ionizes
molecules within the sample, and a detector for detecting the
ionized molecules or fragments of molecules. Non-limiting examples
of common sources include electron impact, electrospray ionization
(ESI), atmospheric pressure chemical ionization (APCI), atmospheric
pressure photo ionization (APPI), matrix assisted laser desorption
ionization (MALDI), surface enhanced laser desorption ionization
(SELDI), and derivations thereof. Common mass separation and
detection systems can include quadrupole, quadrupole ion trap,
linear ion trap, time-of-flight (TOF), magnetic sector, ion
cyclotron (FTMS), Orbitrap, and derivations and combinations
thereof. The advantage of FTMS over other MS-based platforms is its
high resolving capability that allows for the separation of
metabolites differing by only hundredths of a Dalton, many which
would be missed by lower resolution instruments.
[0172] By the term "metabolite", it is meant specific small
molecules, the levels or intensities of which are measured in a
sample, and that may be used as markers to diagnose a disease
state. These small molecules may also be referred to herein as
"metabolite marker", "metabolite component", "biomarker", or
"biochemical marker".
[0173] The metabolites are generally characterized by their
accurate mass, as measured by mass spectormetry technique used in
the above method. The accurate mass may also be referred to as
"accurate neutral mass" or "neutral mass". The accurate mass of a
metabolite is given herein in Daltons (Da), or a mass substantially
equivalent thereto. By "substantially equivalent thereto", it is
meant that a +/-5 ppm difference in the accurate mass would
indicate the same metabolite, as would be recognized by a person of
skill in the art. The accurate mass is given as the mass of the
neutral metabolite. As would be recognized by a person of skill in
the art, the ionization of the metabolites, which occurs during
analysis of the sample, the metabolite will cause either a loss or
gain of one or more hydrogen atoms and a loss or gain of an
electron. This changes the accurate mass to the "ionized mass",
which differs from the accurate mass by the mass of hydrogens and
electrons lost or gained during ionization. Unless otherwise
specified, the accurate neutral mass will be referred to
herein.
[0174] Similarly, when a metabolite is described by its molecular
formula or structure, the molecular formula or structure of the
neutral metabolite will be given. Naturally, the molecular formula
or structure of the ionized metabolite will differ from the neutral
molecular formula or structure by the number of hydrogens lost or
gained during ionization.
[0175] Data is collected during analysis and quantifying data for
one or more than one metabolite is obtained. "Quantifying data" is
obtained by measuring the levels or intensities of specific
metabolites present in a sample.
[0176] The quantifying data is compared to corresponding data from
one or more than one reference sample. The "reference sample" is
any suitable reference sample for the particular disease state. For
example, and without wishing to be limiting in any manner, in the
present invention the reference sample may be a sample from a
non-demented control individual, i.e., a person not suffering from
AD dementia, non-AD dementia or cognitive impairment (also referred
to herein as a "`normal` counterpart"); the reference sample may
also be a sample obtained from a patient with clinically diagnosed
with AD, a patient with clinically diagnosed non-AD dementia, or a
patient diagnosed with significant cognitive impairment. As would
be understood by a person of skill in the art, more than one
reference sample may be used for comparison to the quantifying
data. For example and without wishing to be limiting, the one or
more than one reference sample may be a first reference sample
obtained from a non-demented control individual. The one or more
than one reference sample may further include a second reference
sample obtained from a patient with clinically diagnosed
AD-dementia, a third reference sample obtained from a patient with
clinically diagnosed non-AD dementia, a fourth reference sample
obtained from a patient suffering from significant cognitive
impairment, or any combination thereof.
[0177] The present invention also provides novel compounds,
identified using the methods of the present invention. The novel
compounds may be used as metabolite markers in the differential
diagnosis of dementia, as described above.
[0178] In one embodiment, the compounds may be selected from the
metabolites listed in Tables 1 to 7, or a combination thereof.
These metabolites were identified in serum samples, and may be
phosphatidylcholine-related compounds, ethanolamine plasmalogens,
endogenous fatty acids, essential fatty acids, lipid oxidation
byproducts, metabolite derivatives of said metabolite classes, and
any metabolite that may contribute in any way to the
anabolic/catabolic metabolism of said metabolite classes.
[0179] An optimal panel of compounds may be indentified from those
metabolites listed in Tables 1 to 7. For example and without
wishing to be limiting, the metabolite markers may be metabolites
with accurate masses measured in Daltons of, or substantially
equivalent to, 541.3432, 569.3687, 699.5198, 723.5195, 723.5197,
751.5555, 803.568, 886.5582. The metabolites of accurate masses
699.5198, 723.5195, 723.5197, and 751.555 have presently been
identified as ethanolamine plasmalogens, and are specifically
decreased in patients with AD dementia. The metabolite markers of
accurate masses 541.3432, 569.3687, 803.568, and 886.5582 have
presently been identified as phosphatidylcholine related
metabolites, and are decreased in patients with cognitive
impairment on ADAS-cog, and severity of cognitive impairment
correlates to the degree of decrease.
[0180] The metabolites with accurate masses measured in Daltons of,
or substantially equivalent to, a) 541.3432, b) 569.3687, c)
699.5198, d) 723.5195, e) 751.5555, f) 803.568, can be further
characterized by [0181] a) an extracted ion chromatogram (EIC) as
shown in FIG. 4A, and an MS/MS spectrum as shown in FIG. 6; a
molecular formula of C.sub.25H.sub.51NO.sub.9P; and/or the
structure shown in FIG. 12; [0182] b) an EIC as shown in FIG. 4B,
and an MS/MS spectrum as shown in FIG. 7; a molecular formula of
C.sub.27H.sub.55NO.sub.9P; and/or the structure shown in FIG. 13;
[0183] c) an EIC as shown in FIG. 4C, and an MS/MS spectrum as
shown in FIG. 8; a molecular formula of C.sub.39H.sub.74NO.sub.7P;
and/or the structure shown in FIG. 17; [0184] d) an EIC as shown in
FIG. 4D, and an MS/MS spectrum as shown in FIG. 9; a molecular
formula of C.sub.41H.sub.74NO.sub.7P; and/or the structure shown in
FIG. 18; [0185] e) an EIC as shown in FIG. 4E, and an MS/MS
spectrum as shown in FIG. 10; a molecular formula of
C.sub.43H.sub.78NO.sub.7P; and/or the structure shown in FIG. 19;
[0186] f) an EIC as shown in FIG. 4F, and an MS/MS spectrum as
shown in FIG. 11; a molecular formula of
C.sub.43H.sub.81NO.sub.10P; and/or the structure shown in FIG. 14,
[0187] respectively.
[0188] It is presently shown that the ethanolamine plasmalogen
metabolites (neutral masses 699.5198, 723.5195, 751.5555) and the
phosphatidylcholine metabolites (neutral masses 699.5198, 723.5195,
751.5555) are decreased in the serum of AD subjects exhibiting
significant cognitive impairment. This is the first report of
serum-based changes in these metabolites associated with AD and
dementia. It is further shown that the decrease in the disclosed
serum phospatidylcholine related metabolites occurs in all patients
exhibiting a significant cognitive impairment as measured by the
ADAS-cog regardless of AD status, and that the degree of decrease
correlates with the severity of the cognitive impairment. However,
the observed decrease in disclosed ethanolamine plasmalogens is
independent of cognitive impairment, occurs specifically in
subjects with AD and is therefore a true diagnostic of AD.
[0189] Ethanolamine plasmalogens are a type of ethanolamine
phospholipid. Ethanolamine phospholipids can be further
differentiated based on their sn-1 configurations (either acyl,
ether, or vinyl ether). The sn-2 position is typically acyl and the
sn-3 position contains the phosphoethanolamine moiety. Therefore,
the three classes are described as either diacyl (also referred to
herein as PtdEt), alkyl-acyl (also referred to herein as plasmanyl)
or alkenyl-acyl (also referred to herein as EtnPl or plasmenyl).
Various basic structures of ethanolamine phospholipids are shown in
FIG. 20, along with the standard naming convention used herein.
[0190] A decrease in the disclosed ethanolamine plasmalogens may
represent the initial or early stages AD, and can be detected
non-invasively in living subjects by measuring serum levels of
specific ethanolamine plasmalogens. Similarly, cognitive impairment
can be quantitated non-invasively by measuring the serum levels of
specific phosphatidylcholine metabolites.
[0191] Other metabolites have also been identified. For example,
the metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, a) 567.3547, b) 565.3394, c) 805.5832,
d) 827.57, e) 829.5856, f) 831.5997, and g) 853.5854, which may be
further characterized by [0192] a) the molecular formula
C.sub.27H.sub.55NO.sub.9P; and/or the structure shown in FIG. 15A
[0193] b) the molecular formula C.sub.27H.sub.55NO.sub.9P; and/or
the structure shown in FIG. 15B [0194] c) the molecular formula
C.sub.43H.sub.83NO.sub.10P; and/or the structure shown in FIG. 15C;
[0195] d) the molecular formula C.sub.45H.sub.81NO.sub.10P; and/or
the structure shown in FIG. 15D; [0196] e) the molecular formula
C.sub.45H.sub.83NO.sub.10P; and/or the structure shown in FIG. 15E;
[0197] f) the molecular formula C.sub.45H.sub.85NO.sub.10P; and/or
the structure shown in FIG. 15F; [0198] g) the molecular formula
C.sub.47H.sub.83NO.sub.10P; and/or the structure shown in FIG. 15G,
[0199] respectively.
[0200] Based on the identification of metabolites specific to AD
dementia (accurate masses 699.5198, 723.5195, 723.5197, 751.555) as
ethanolamine plasmalogens, other ethanolamine phospholipid
metabolite markers were identified. These are metabolites M05 to
M24 as listed and characterized (accurate mass, name/composition,
molecular formula) in Table 18. The structure of the metabolite can
be deduced based on the metabolite name as indicated in Table 18
and the nomenclature, as indicated in FIG. 20.
[0201] Of the compounds listed in Table 18, those of particular
interest include metabolites with accurate masses measured in
Daltons of, or substantially equivalent to, a) 701.53591, b)
699.52026, c) 723.52026, d) 747.52026, e) 729.56721, f) 727.55156,
g) 779.58286, and h) 775.55156, which can be further characterized
by [0202] a) a MS/MS spectrum as shown in FIG. 21; molecular
formula C.sub.27H.sub.55NO.sub.9P; and/or the structure
[0202] ##STR00005## [0203] b) a MS/MS spectrum as shown in FIG. 22;
molecular formula C.sub.39H.sub.74NO.sub.7P; and/or the
structure
[0203] ##STR00006## [0204] c) a MS/MS spectrum as shown in FIG. 23;
molecular formula C.sub.41H.sub.74NO.sub.7P; and/or the
structure
[0204] ##STR00007## [0205] d) a MS/MS spectrum as shown in FIG. 24;
molecular formula C.sub.43H.sub.74NO.sub.7P; and/or the
structure
[0205] ##STR00008## [0206] e) a MS/MS spectrum as shown in FIG. 25;
molecular formula C.sub.41H.sub.80NO.sub.7P; and/or the
structure
[0206] ##STR00009## [0207] f) a MS/MS spectrum as shown in FIG. 26;
molecular formula C.sub.41H.sub.78NO.sub.7P; and/or the
structure
[0207] ##STR00010## [0208] g) a MS/MS spectrum as shown in FIG. 27;
molecular formula C.sub.45H.sub.82NO.sub.7P; and/or the
structure
[0208] ##STR00011## [0209] h) a MS/MS spectrum as shown in FIG. 28;
molecular formula C.sub.45H.sub.78NO.sub.7P; and/or the
structure
[0209] ##STR00012## [0210] respectively.
[0211] In another embodiment of the present invention, the
compounds may be selected from the metabolites listed in Table 13,
or a combination thereof. These metabolites were identified in CSF
samples. Of particular interest are the metabolites with accurate
masses measured in Daltons of, or substantially equivalent to,
207.0822, 275.8712, 371.7311, 373.728, 432.1532, 485.5603,
487.6482, 562.46, 622.2539, 640.2637, 730.6493, 742.2972. When used
to diagnose dementia, the metabolite markers 207.0822, 432.1532,
562.46, 622.2539, 640.2637, 730.6493, and 742.2972 are increased in
patients with AD dementia; and metabolite markers 275.8712,
371.7311, 373.728, 485.5603, and 487.6482 are decreased in patients
with AD dementia.
[0212] In a further method of the present invention, a method for
assessing dementia or the risk of dementia in a patient is
described. The method comprises the steps of: [0213] a) obtaining a
serum sample from said patient; [0214] b) analyzing said sample to
obtain quantifying data for one or more than one metabolite marker;
[0215] c) comparing the quantifying data for said one or more than
one metabolite marker to corresponding data obtained from one or
more than one reference sample; and [0216] d) using said comparison
to assess dementia or the risk of dementia.
[0217] The step of analyzing the sample (steb b)) may comprise
analyzing the sample by liquid chromatography mass spectrometry
(LC-MS). Alternatively, the step of analyzing the sample (step b))
may comprise analyzing the sample by linear ion trap mass
spectrometry followed by liquid chromatograph, when the method is a
highthroughput method.
[0218] The one or more than one reference sample may include a
first reference sample obtained from a non-demented control
individual, a second reference sample obtained from a patient with
cognitive impairment as measured by ADAS-cog, a third reference
sample obtained from a patient with cognitive impairment as
measured by MMSE, or a combination of one or more of these.
[0219] Without wishing to be limiting in any manner, the one or
more than one metabolite marker used to asses dementia or the risk
of dementia may be selected from the metabolites listed in Tables
10-12, or a combination thereof. Of particular interest are
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to 541.3432, 569.3687, 699.5198, 723.5195,
723.5197, 751.5555, 803.568, 886.5582, 565.3394, 569.369, 801.555,
857.6186. A decrease in the patient sample in metabolite markers
699.5198, 723.5195, 723.5197, and 751.555 indicates AD pathology; a
decrease in the patient sample in metabolite markers 541.3432,
569.3687, 803.568, and 886.5582 indicates cognitive impairment on
ADAS-cog; and 565.3394, 569.369, 801.555, and 857.6186 indicates
cognitive impairment on MMSE.
[0220] In yet another embodiment of the present invention, there is
provided a method for differentially diagnosing dementia or the
risk of dementia in a patient. The method comprising the steps of:
[0221] a) obtaining a sample from said patient; [0222] b) analyzing
said sample to obtain quantifying data for one or more than one
metabolite marker; [0223] c) obtaining a ratio for each of the one
or more than one metabolite marker to an internal control
metabolite; [0224] d) comparing each ratio of said one or more than
one metabolite marker to the internal control metabolite to
corresponding data obtained from one or more than one reference
sample; and [0225] e) using said comparison to differentially
diagnose dementia or the risk of dementia.
[0226] The step of analyzing the sample (steb b) may comprise
analyzing the sample using a mass spectrometer (MS). For example,
and without wishing to be limiting, such mass spectrometer could be
of the FTMS, orbitrap, time of flight (TOF) or quadrupole types.
Alternatively, the mass spectrometer could be equipped with an
additional pre-detector mass filter. For example, and without
wishing to be limiting such instruments are commonly referred to as
quadrupole-FTMS (Q-FTMS), quadrupole-TOF (Q-TOF) or triple
quadrupole (TQ or QQQ). In addition, the mass spectrometer could be
operated in either the parent ion detection mode (MS) or in MSn
mode, where n>=2. MSn refers to the situation where the parent
ion is fragmented by collision induced dissociation (CID) or other
fragmentation procedures to create fragment ions, and then one or
more than one of said fragments are detected by the mass
spectrometer. Such fragments can then be further fragmented to
create further fragments. Alternatively, the sample could be
introduced into the mass spectrometer using a liquid or gas
chromatographic system or by direct injection.
[0227] In the method as just described above, the one or more than
one reference sample may be a first reference sample obtained from
a non-demented control individual. The one or more than one
reference sample may further include a second reference sample
obtained from a patient with clinically diagnosed AD-dementia, a
third reference sample obtained from a patient with clinically
diagnosed non-AD dementia, a fourth reference sample obtained from
a patient suffering from significant cognitive impairment, or any
combination thereof.
[0228] In the method as described above, the sample and reference
sample may be serum samples. The one or more than one metabolite
marker may be selected from the metabolites as listed and
characterized (accurate mass, name/composition, molecular formula)
in Table 18. The "internal control metabolite" refers to an
endogenous metabolite naturally present in the patient. Any
suitable endogenous metabolite that does not vary over the disease
states can be used as the internal control metabolite. For example,
and without wishing to be limiting, the internal control metabolite
may be phosphatidylethanolamine 16:0/18:0 (PtdEt 16:0/18:0, M01),
as shown in Table 18; this internal control metabolite has a
molecular formula of C.sub.19H.sub.78NO.sub.8P, and a structure
characterized as
##STR00013##
[0229] Use of the ratio of the metabolite marker to the internal
control metabolite offers measurement that are more stable and
reproducible than measurement of absolute levels of the metabolite
marker. As the internal control metabolite is naturally present in
all samples and does not appear to vary significantly over disease
states, the sample-to-sample variability (due to handling,
extraction, etc) is minimized.
[0230] Of the compounds listed in Table 18, those of particular
interest in the above method include metabolites with accurate
masses measured in Daltons of, or substantially equivalent to, a)
701.53591, b) 699.52026, c) 723.52026, d) 747.52026, e) 729.56721,
f) 727.55156, g) 779.58286, and h) 775.55156. A decrease in the
ratio of a) to h) to the internal control metabolite indicates AD
dementia with a severe cognitive impairment. These metabolites can
be further characterized by [0231] a) a MS/MS spectrum as shown in
FIG. 21; molecular formula C.sub.27H.sub.55NO.sub.9P; and/or the
structure
[0231] ##STR00014## [0232] b) a MS/MS spectrum as shown in FIG. 22;
molecular formula C.sub.39H.sub.74NO.sub.7P; and/or the
structure
[0232] ##STR00015## [0233] c) a MS/MS spectrum as shown in FIG. 23;
molecular formula C.sub.41H.sub.74NO.sub.7P; and/or the
structure
[0233] ##STR00016## [0234] d) a MS/MS spectrum as shown in FIG. 24;
molecular formula C.sub.43H.sub.74NO.sub.7P; and/or the
structure
[0234] ##STR00017## [0235] e) a MS/MS spectrum as shown in FIG. 25;
molecular formula C.sub.41H.sub.80NO.sub.7P; and/or the
structure
[0235] ##STR00018## [0236] f) a MS/MS spectrum as shown in FIG. 26;
molecular formula C.sub.41H.sub.78NO.sub.7P; and/or the
structure
[0236] ##STR00019## [0237] g) a MS/MS spectrum as shown in FIG. 27;
molecular formula C.sub.45H.sub.82NO.sub.7P; and/or the
structure
[0237] ##STR00020## [0238] h) a MS/MS spectrum as shown in FIG. 28;
molecular formula C.sub.45H.sub.78NO.sub.7P; and/or the
structure
[0238] ##STR00021## [0239] respectively.
[0240] In yet another embodiment of the present invention, there is
provided a method for evaluating the efficacy of a therapy for
treating dementia in a patient, comprising: [0241] a) obtaining a
sample from said patient; [0242] b) analyzing said sample to obtain
quantifying data for one or more than one metabolite marker; [0243]
c) comparing said quantifying data to corresponding data obtained
from one or more than one reference sample; and [0244] d) using
said comparison to determine whether the therapy is improving the
demented state of the patient.
[0245] Optionally, after the step of analyzing (step b), a ratio
for each of the one or more than one metabolite marker to an
internal control metabolite may be obtained. In this case, each
ratio of said one or more than one metabolite marker to the
internal control metabolite to corresponding data obtained from one
or more than one reference sample is compared to evaluate the
efficacy of the therapy.
[0246] The step of analyzing (step b) may comprise analyzing the
sample by liquid chromatography mass spectrometry (LC-MS), or
alternatively may comprise analyzing the sample by liquid
chromatography and linear ion trap mass spectrometry when the
method is a highthroughput method.
[0247] By the term "therapy", it is meant any suitable course of
therapy that may improve the health state or demented state of the
patient being evaluated. When evaluating the efficacy of the
therapy, the effect of the particular therapy in improving or
degrading the health state of the patient will be measured. In
doing so, a person of skill in the art would be capable of
determining whether the therapy is effective for treating the
demented state.
[0248] In the methods as described, the one or more than one
reference sample may be any suitable reference sample. For example,
and without wishing to be limiting in any manner, the reference
sample may be a plurality of samples obtained from non-demented
control individuals; a plurality of samples obtained from
clinically diagnosed AD patients; one or more than one pre-therapy
baseline sample obtained from the patient; or any combination
thereof. A pre-therapy baseline sample from the patient is
particularly useful, as the variation in metabolites will then be
specific to the patient.
[0249] The sample and the reference sample may be serum samples. In
this case, the one or more than one metabolite marker could be
selected from the metabolites listed in Tables 1 to 7, or a
combination thereof, for example, metabolite markers with accurate
masses measured in Daltons of, or substantially equivalent to,
541.3432, 569.3687, 699.5198, 723.5195, 723.5197, 751.5555,
803.568, 886.5582. Alternatively, the metabolite markers may be
selected from metabolites M05 to M24 with accurate masses of, or
substantially equivalent to those listed in Table 18, for example,
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, 701.53591, 699.52026, 723.52026,
747.52026, 729.56721, 727.55156, 779.58286, and 775.55156.
Metabolites M05-M24 could also be used when a ratio is obtained
between the metabolites and the internal control metabolite; the
internal metabolite could be, for example, metabolite M01, as
described in Table 18.
[0250] The sample and the reference sample may also be
cerebrospinal fluid (CSF) samples. In this case, the one or more
than one metabolite marker could be selected from the metabolites
listed in Table 13, or a combination thereof; for example,
metabolites with accurate masses measured in Daltons of, or
substantially equivalent to, 207.0822, 275.8712, 371.7311, 373.728,
432.1532, 485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493,
742.2972.
[0251] The identified metabolites can be readily measured
systemically. This point is of fundamental importance, since the
majority of research pertaining to AD and other neurological
disorders has ignored the peripheral systems. The ability to
measure neurodegenerative processes within a blood sample is of
substantial value in the diagnosis of dementia. With respect to the
specific ethanolamine plasmalogen metabolites of the present
invention, these are a valid biochemical marker of AD pathology
since this molecular species' content does not change in
Parkinson's disease, a disease which is often accompanied by
dementia [29]. Furthermore, the specificity of the plasmalogen
metabolites to AD indicates that its levels in serum could be
readily measured longitudinally throughout the lifetime of an
individual to assess the risk or for early detection of the disease
prior to the emergence of clinical symptoms.
[0252] The present invention also provides high throughput methods
for differential diagnosis of AD dementia and non-AD dementia
states. The method may involve fragmentation of the parent
molecule; in a non-limiting example, this may be accomplished by a
Q-Trap.TM. system. Detection of the metabolites may be performed
using one of various assay platforms, including colorimetric
chemical assays (UV, or other wavelength), antibody-based
enzyme-linked immunosorbant assays (ELISAs), chip-based and
polymerase-chain reaction for nucleic acid detection assays,
bead-based nucleic-acid detection methods, dipstick chemical assays
or other chemical reaction, image analysis such as magnetic
resonance imaging (MRI), positron emission tomography (PET) scan,
computerized tomography (CT) scan, nuclear magnetic resonance
(NMR), and various mass spectrometry-based systems.
[0253] A high-throughput method for determining the levels of the
metabolites in a person's blood and comparing the levels to levels
in a normal "reference" population can lead to a prediction of
whether the person has AD or not. This can be carried out in
several ways. One way is to use a prediction algorithm to classify
the test sample, as previously described, which would output a
percentage probability for having AD. A predictive approach would
work independently of the assay method, as long as the intensities
of the metabolites could be measured. Another method could simply
be based on setting a threshold intensity level from the mass
spectrometer, and determining whether a person's profile is above
or below the threshold which would indicate their AD status.
Alternatively, and without wishing to be limiting in any manner, a
preferred method is a truly quantitative assay could be performed
to determine the molar concentration of the six metabolites in the
non-demented normal and AD population. An absolute threshold
concentration could then be determined for AD-positivity. In a
clinical setting, this would mean that if the measured levels of
the metabolites, or combinations of the metabolites, were below a
certain concentration, there would be an associated probability
that the individual is positive for AD. Therefore, the optimal
diagnostic test could comprise a method of measuring the
intensities of the metabolites in serum, and an algorithm for
taking the intensity values and outputting a predicted probability
for having AD as well as for being healthy (i.e., AD-negative).
[0254] The methods and identified biomarkers of the present
invention, based on small molecules or metabolites in a sample,
fulfills the criteria identified in 1999 for an ideal screening
test [82], as development of assays capable of detecting specific
metabolites is relatively simple and cost effective per assay. The
test is minimally invasive and is indicative of cognitive
impairment and of AD pathology. Translation of the method into a
clinical assay compatible with current clinical chemistry
laboratory hardware is commercially acceptable and effective.
Furthermore, the method of the present invention does not require
highly trained personnel to perform and interpret the test.
[0255] The present invention will be further illustrated in the
following examples.
Example 1
Identification of Differentially Expressed Metabolites
[0256] Differentially expressed metabolites were identified in
clinically diagnosed AD with and without significant cognitive
impairment, clinically diagnosed non-AD and non-demented
controls.
[0257] Clinical Samples.
[0258] For the AD serum diagnostic assay described, samples were
obtained from representative populations of non-demented healthy
individuals and of clinically diagnosed AD and non-AD dementia
patients. The biochemical markers of AD described in the invention
were derived from the analysis of 75 serum samples from patients
clinically diagnosed with probable AD (43 patients with significant
cognitive impairment, 32 with no cognitive impairment), serum
samples from 30 patients with clinically diagnosed non-AD dementia,
and 31 serum samples from non-demented controls. Samples in the
three groups were from a diverse population of individuals, ranging
in age, ethnicity, weight, occupation, and displaying varying
non-dementia-related health-states. All samples were single
time-point collections. Cognitive impairment of the patients was
also assessed using the Alzheimer's Disease Assessment Scale
(ADAS)-cognitive subset.
[0259] For the AD CSF diagnostic assay described, samples were
obtained from a group of patients that represented clinically
diagnosed AD with dementia and non-AD patients with dementia. The
biochemical markers of AD described in this invention were derived
from the analysis of 6 CSF samples from clinically diagnosed AD
patients with dementia and 5 CSF samples from clinically diagnosed
non-AD patients with dementia.
[0260] Samples in both groups were from a diverse population of
individuals, ranging in age, ethnicity, weight, occupation, and
displaying varying non-dementia-related health-states. All samples
were single time-point collections. The metabolites contained
within the 136 serum samples and 11 CSF samples used in this
application were separated into polar and non-polar extracts
through sonication and vigorous mixing (vortex mixing).
[0261] Mass Spectrometry Analysis.
[0262] Analysis of serum extracts collected from 136 individuals
(75 clinically diagnosed AD, 30 clinically diagnosed non-AD, and 31
non-demented healthy controls) and 11 CSF extracts (6
clinically-diagnosed AD and 5 clinically diagnosed non-AD patients)
was performed by direct injection into a FTMS and ionization by
either ESI or atmospheric pressure chemical ionization (APCI) in
both positive and negative modes. Sample extracts were diluted
either three or six-fold in methanol:0.1% (v/v) ammonium hydroxide
(50:50, v/v) for negative ionization modes, or in methanol:0.1%
(v/v) formic acid (50:50, v/v) for positive ionization modes. For
APCI, sample extracts were directly injected without diluting. All
analyses were performed on a Bruker Daltonics APEX III Fourier
transform ion cyclotron resonance mass spectrometer equipped with a
7.0 T actively shielded superconducting magnet (Bruker Daltonics,
Billerica, Mass.). Samples were directly injected using
electrospray ionization (ESI) and/or APCI at a flow rate of 1200
.mu.L per hour. Ion transfer/detection parameters were optimized
using a standard mix of serine, tetra-alanine, reserpine,
Hewlett-Packard tuning mix and the adrenocorticotrophic hormone
fragment 4-10. In addition, the instrument conditions were tuned to
optimize ion intensity and broad-band accumulation over the mass
range of 100-1000 amu according to the instrument manufacturer's
recommendations. A mixture of the abovementioned standards was used
to internally calibrate each sample spectrum for mass accuracy over
the acquisition range of 100-1000 amu.
[0263] In total, six separate analyses comprising combinations of
extracts and ionization modes were obtained for each sample:
[0264] Aqueous Extract [0265] 1. Positive ESI (analysis mode 1101)
[0266] 2. Negative ESI (analysis mode 1102)
[0267] Organic Extract [0268] 3. Positive ESI (analysis mode 1201)
[0269] 4. Negative ESI (analysis mode 1202) [0270] 5. Positive APCI
(analysis mode 1203) [0271] 6. Negative APCI (analysis mode
1204)
[0272] Mass Spectrometry Data Processing.
[0273] Using a linear least-squares regression line, mass axis
values were calibrated such that each internal standard mass peak
had a mass error of <1 p.p.m. compared with its theoretical
mass. Using XMASS software from Bruker Daltonics Inc., data file
sizes of 1 megaword were acquired and zero-filled to 2 megawords. A
sinm data transformation was performed prior to Fourier transform
and magnitude calculations. The mass spectra from each analysis
were integrated, creating a peak list that contained the accurate
mass and absolute intensity of each peak. Compounds in the range of
100-2000 m/z were analyzed. In order to compare and summarize data
across different ionization modes and polarities, all detected mass
peaks were converted to their corresponding neutral masses assuming
hydrogen adduct formation. A self-generated two-dimensional (mass
vs. sample intensity) array was then created using
DISCOVAmetrics.TM. software (Phenomenome Discoveries Inc.,
Saskatoon, SK, Canada). The data from multiple files were
integrated and this combined file was then processed to determine
the unique masses. The average of each unique mass was determined,
representing the y axis. This value represents the average of all
of the detected accurate masses that were statistically determined
to be equivalent. Considering that the mass accuracy of the
instrument for the calibration standards is approximately 1 ppm, a
person skilled in the art will recognize that these average masses
may include individual masses that fall within +/-5 ppm of this
average mass. A column was created for each file that was
originally selected to be analyzed, representing the x axis. The
intensity for each mass found in each of the files selected was
then filled into its representative x,y coordinate. Coordinates
that did not contain an intensity value were left blank. Once in
the array, the data were further processed, visualized and
interpreted, and putative chemical identities were assigned. Each
of the spectra were then peak picked to obtain the mass and
intensity of all metabolites detected. These data from all of the
modes were then merged to create one data file per sample. The data
from all 136 samples was then merged and aligned to create a
two-dimensional metabolite array in which each sample is
represented by a column and each unique metabolite is represented
by a single row. In the cell corresponding to a given metabolite
sample combination, the intensity of the metabolite in that sample
is displayed. When the data is represented in this format,
metabolites showing differences between groups of samples were
determined. The same procedure was utilized to combine the 11 CSF
samples in a two-dimensional metabolite array.
[0274] A. Serum Biomarkers
[0275] A student's T-test was used to select for metabolites which
differed significantly between the following different clinical
groups in serum. Metabolites that were less than p<0.05 were
considered significant.
[0276] A1--Clinically diagnosed AD patients (n=75) vs. non-demented
controls (n=31). This comparison yielded 262 metabolites (see Table
1).
[0277] A2--Clinically diagnosed AD patients with a significant
cognitive impairment (n=32) vs. non-demented controls (n=31). This
comparison yielded 292 metabolites (see Table 2).
[0278] A3--Clinically diagnosed AD patients with a significant
cognitive impairment (n=32) vs. clinically diagnosed non-AD
patients with a significant cognitive impairment (n=30); this
comparison yielded 118 metabolites markers (see Table 3).
[0279] A4--Clinically diagnosed AD patients with significant
cognitive impairment (n=32) vs. clinically diagnosed AD patients
without significant cognitive impairment (n=43). This comparison
yielded 97 metabolites markers (see Table 4).
[0280] A5--Clinically diagnosed non-AD patients (n=30) vs.
non-demented controls (n=31); this comparison yielded 199
metabolites markers (see Table 5).
[0281] A6--Clinically diagnosed AD patients with mild cognitive
impairment (n=43) vs. non-demented controls (n=31). This comparison
yielded 136 metabolites (see Table 6).
[0282] A7--Patients with significant cognitive impairment (n=42)
and patients with a mild cognitive impairment (n=43). This
comparison yielded 81 metabolites (Table 7).
[0283] Tables 1-7 show biochemical markers whose concentrations or
amounts in serum are significantly different (p<0.05) between
the tested populations and therefore have potential diagnostic
utility for identifying each of the aforesaid populations. The
features are described by their accurate mass and analysis mode,
which together are sufficient to provide the putative molecular
formulas and chemical characteristics (such as polarity and
putative functional groups) for each metabolite.
[0284] From the initial lists of several hundred possible
metabolites, it was determined that a combination of 8 metabolites
fulfills the criteria for a serum dementia test: the combination
can differentiate AD dementia from non-AD dementia, the early
stages of AD and healthy controls. The best combination of 8
metabolites included the metabolites with neutral masses (measured
in Daltons) 541.3432, 569.3687, 699.5198, 723.5195, 723.5197,
751.5555, 803.568, 886.5582. Although these are the actual masses,
a person skilled in the art of this technology would recognize that
+/-5 ppm difference would indicate the same metabolite.
[0285] In analyzing the present results, a person of skill in the
art would understand that the following clinical groups are of
interest: non-AD with significant cognitive impairment, AD with
significant cognitive impairment, AD without significant cognitive
impairment and non-demented controls. Bar graphs representing the
mean+/-SEM of the 8 biomarkers for the four different clinical
groups are shown in FIG. 1. Relative to control, non-demented
individuals, the three non-control states can be described as
follows:
[0286] 1. Non-AD with significant cognitive impairment vs. control:
[0287] a. Biomarker 541.3432--decreased [0288] b. Biomarker
569.3687--decreased [0289] c. Biomarker 699.5198--no difference
[0290] d. Biomarker 723.5195--no difference [0291] e. Biomarker
723.5197--no difference [0292] f. Biomarker 751.5555--no difference
[0293] g. Biomarker 803.568--decreased [0294] h. Biomarker
886.5582--decreased
[0295] 2. Clinically diagnosed AD with significant cognitive
impairment vs. control [0296] a. Biomarker 541.3432--decreased
[0297] b. Biomarker 569.3687--decreased [0298] c. Biomarker
699.5198--decreased [0299] d. Biomarker 723.5195--decreased [0300]
e. Biomarker 723.5197--decreased [0301] f. Biomarker
751.5555--decreased [0302] g. Biomarker 803.568--decreased [0303]
h. Biomarker 886.5582--decreased [0304] 3. Clinically diagnosed AD
without significant cognitive impairment vs. control [0305] a.
Biomarker 541.3432--decreased [0306] b. Biomarker 569.3687--no
difference [0307] c. Biomarker 699.5198--decreased [0308] d.
Biomarker 723.5195--decreased [0309] e. Biomarker
723.5197--decreased [0310] f. Biomarker 751.5555--decreased [0311]
g. Biomarker 803.568--no difference [0312] h. Biomarker
886.5582--no difference
[0313] In each of the three non-control cases described above, a
unique subset of markers was decreased.
[0314] Bar graphs representing the mean+/-SEM of the 8 biomarkers
for the two different clinical groups with a significant cognitive
impairment are shown in FIG. 2. Relative to non-AD dementia with
significant cognitive impairment, AD patients with significant
cognitive impairment can be described as: [0315] a. Biomarker
541.3432--no difference [0316] b. Biomarker 569.3687--no difference
[0317] c. Biomarker 699.5198--decreased [0318] d. Biomarker
723.5195--decreased [0319] e. Biomarker 723.5197--decreased [0320]
f. Biomarker 751.5555--decreased [0321] g. Biomarker 803.568--no
difference [0322] h. Biomarker 886.5582--no difference
[0323] The results of this invention show a clear distinction
between the serum of individuals with clinically diagnosed AD WITH
a significant cognitive impairment, individuals with clinically
diagnosed AD WITHOUT a significant cognitive impairment (this could
be early stage AD), individuals with non-AD dementia WITH a
significant cognitive impairment, and non-demented controls. These
findings are capable of identifying and distinguishing the
different types of dementia from one another and from the early
stages of cognitive impairment as described in this application.
From the above results, it can be further concluded that the
metabolite markers with masses 699.5198, 723.5195, 723.5997,
751.5555 are specific for AD pathology; while markers with masses
of 541.3432, 569.3687, 803.568, 886.5582 are specific for cognitive
impaired based on ADAS-cog testing.
[0324] A second neuropsychological test, Folstein's Mini-Mental
State Exam (MMSE), which measures cognitive impairment, was applied
to all 136 patients. The MMSE is widely used and is an extensively
validated test of orientation, short and long-term memory, praxis,
language and comprehension. In the clinically diagnosed AD patients
that had no significant cognitive impairment (n=43), 15 of those
patients had a score on MMSE that would indicate normal cognition
(MMSE.gtoreq.28), whereas the remaining 28 patients had MMSE scores
that indicated a mild impairment (score 18-23, n=11) or severe
cognitive impairment (score 9-17, n=17). A F-test was used to
select for metabolites which differed significantly between the
MMSE scores (normal, mild or severe cognitive impairment) for 43
clinically diagnosed AD patients with no significant cognitive
impairment on the ADAS-cog test (p<0.05). 23 metabolites met
this criterion (shown in Table 8). These are all features which
differ statistically between the two populations and therefore have
potential diagnostic utility. The features are described by their
accurate mass and analysis mode, which together are sufficient to
provide the putative molecular formulas and chemical
characteristics (such as polarity and putative functional groups)
of each metabolite.
[0325] An optimal subset of 4 metabolites, all of which were
observed to decrease, from the 23 metabolites was selected using
Principal Components Analysis (PCA). The 4 metabolites able to
produce the greatest separation between the groups were 565.3394,
569.369, 801.555, 857.6186. The metabolites are indicated by
asterisks on Table 8 and represent a 4-metabolite biomarker panel
associated with cognitive impairment on MMSE. The fact that a
second set of metabolites were associated cognitive impairment
suggests that the MMSE must be specific to one or several other
cognitive states that the ADAS-cog is not specifically
measuring.
[0326] Therefore, a total of three 4-biomarker panels can be
applied to the 136 patients to classify them into one of 8
categories which will simultaneously indicate the presence of AD
pathology (biomarkers 699.5198, 723.5195, 723.5997, 751.5555),
cognitive impaired on ADAS-cog (541.3432, 569.3687, 803.568,
886.5582) and cognitive impaired on MMSE (565.3394, 569.369,
801.555, 857.6186). Using a 0/1 binary model, each patient can be
labeled using a 3 digit code from "000" indicating no cognitive
impairment and no AD pathology to "111" indicating both MMSE and
ADAS-cog impairment and AD pathology. Table 9 indicates the
separation of the patient samples into the 8 categories.
[0327] The three 4-biomarker panels were applied individually to
the metabolite array and the patients that showed the best
separation on the PCA plot were selected. These patients were
selected because they represented the best discriminator between
the 3 different groups [AD (n=20) vs. non-AD pathology (n=20), high
ADAS score (n=20) vs. low ADAS score (n=12), impaired cognition on
the MMSE score (n=20) vs. normal cognition on the MMSE score
(n=20)]. A student's t-test was performed between the different
clinical groups (p<0.05). The 116 metabolites that met the
p-value criteria for AD vs. non-AD pathology are listed in Table
10. Table 11 lists the 124 metabolites that met the p-value
criteria for high ADAS score vs. low ADAS score, and Table 12
contains the list of 344 metabolites that met the p-value criteria
for impaired score on MMSE and normal cognition on MMSE.
[0328] Both the ADAS-cog and MMSE neuropsychological tests measure
cognitive errors related to praxis, orientation, memory and
language ability. Therefore, it would be reasonable to suggest
biomarkers associated with ADAS-cog score and/or MMSE are related
to the ability to conceive of, organize and initiate unfamiliar
sequences, the awareness of one's self and environment, as well as
memory and language ability. As such, these biomarkers are not
exclusive to cognitive impairment associated with dementia; rather
any condition that results in any type of praxis, orientation,
memory and/or language deficit would show a similar reduction
within a biological sample.
[0329] The sample set (136 individuals) used for this discovery was
not trivial, and was comprised of individuals of various ethnic and
geographical backgrounds, and of varying age and health status.
Therefore, there is sound reason to expect that the findings are
representative of the general dementia population.
[0330] B. CSF Biomarkers.
[0331] A student's T-test was used to select for metabolites which
differ between the clinically diagnosed AD patients and clinically
diagnosed non-AD patients in CSF samples (p<0.05). 42
metabolites met this criterion (shown in Table 13). These
metabolites differed statistically between the two populations and
therefore have potential diagnostic utility. The metabolites are
described by their accurate mass and analysis mode, which together
are sufficient to provide the putative molecular formulas and
chemical characteristics (such as polarity and putative functional
groups) of each metabolite.
[0332] An optimal subset of 12 metabolites from the 42 metabolites
described above was selected. These metabolites had the greatest
statistical difference between the two groups (p<0.01).
Metabolites were excluded if they were not detected in at least 60%
of the samples in each group (4/6 clinically diagnosed AD and 3/5
clinically diagnosed non-AD). The panel comprises masses 207.0822,
275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482, 562.46,
622.2539, 640.2637, 730.6493, 742.2972. Although these are the
actual masses, a person skilled in the art of this technology would
recognize that an +/-5 ppm difference would indicate the same
metabolite.
[0333] The 12 biomarker panel was tested using 5 CSF samples from
undiagnosed patients. The only information available on the samples
was the subject's age, gender, and whether an individual had a
cognitive deficit. If the 12 biomarker panel was correct, the
subject could be diagnosed as having AD dementia, non-AD dementia,
or normal. From the 5 CSF samples provided by undiagnosed patients,
1 was diagnosed with non-AD dementia, 2 with AD dementia, and 2 as
normal. The two normal subjects did not have a cognitive impairment
as indicated by the Mini Mental State Examination (MMSE) score.
Therefore, using a 12 metabolite feature set it was possible to
both diagnose AD and non-AD dementia.
[0334] Bar graphs representing the mean+/-SEM of the 12 biomarkers
for the two different clinical groups are shown in FIG. 3. Relative
to non-AD dementia with significant cognitive impairment, AD
patients with a significant cognitive impairment can be described
as: [0335] a. Biomarker 207.0822--increased [0336] b. Biomarker
275.8712--decreased [0337] c. Biomarker 371.7311--decreased [0338]
d. Biomarker 373.728--decreased [0339] e. Biomarker
432.1532--increased [0340] f. Biomarker 485.5603--decreased [0341]
g. Biomarker 487.6482--decreased [0342] h. Biomarker
562.46--increased [0343] i. Biomarker 622.2539--increased [0344] j.
Biomarker 640.2637--increased [0345] k. Biomarker
730.6493--increased [0346] l. Biomarker 742.2972--increased
[0347] Based on these results, a clear distinction was made between
the CSF of clinically diagnosed non-AD and AD patients. Therefore,
such findings are capable of identifying and distinguishing AD
dementia from non-AD dementia and can form the basis of a dementia
diagnostic test in CSF as described in this application. It is
expected that the finding are representative of the general
dementia population.
[0348] Although a non-targeted FTMS-based platform was used in the
identification and selection of the optimal metabolites in serum
and CSF, other methods of subsequently detecting the molecules,
including other MS-based platforms, ELISAs, colorimetric assays,
etc can used to detect the molecules.
Example 2
Independent Method Confirmation of Discovered Metabolites
[0349] A. Serum Biomarkers
[0350] An independent mass spectrometry method was used to verify
the intensity differences between non-demented normal and
clinically-diagnosed AD serums of the eight diagnostic metabolites
discovered using the FTMS method. Eight representative
clinically-diagnosed AD sample extracts and eight representative
non-demented control sample extracts were analyzed by LC-MS using
an HP 1100 high-performance liquid chromatography interfaced to an
ABI Q-Star mass spectrometer.
[0351] Aqueous fractions from five clinically-diagnosed AD and five
non-demented control sample extracts were evaporated under nitrogen
gas and reconstituted in 100 uL of methanol:water:formic acid
(5:94.9:0.1). Five .mu.L of the reconstituted sample was subjected
to HPLC (Agilent Technologies) (HP 1100 with Metasil AQ 3u,
100.times.2 mm column) for full scan and 10 .mu.L for MS/MS at a
flow rate of 0.2 ml/min.
[0352] Eluate from the HPLC was analyzed using an ABI Q-Star XL
mass spectrometer fitted with a Turboion spray ion (ESI) source in
negative mode. The scan type in full scan mode was time-of-flight
(TOF) with an accumulation time of 1.0000 seconds, mass range
between 50 and 1500 Da, and duration time of 70 min. Source
parameters were as follows: Ion source gas 1 (GS1) 55; Ion source
gas 2 (GS2) 90; Curtain gas (CUR) 40; Nebulizer Current (NC) 0;
Temperature 450.degree. C.; Declustering Potential (DP)-55;
Focusing Potential (FP)-265; Declustering Potential 2 (DP2)-15. In
MS/MS mode, scan type was product ion, accumulation time was 1.0000
seconds, scan range between 50 and 1000 Da and duration time 70
min. All source parameters are the same as above, with a collision
energy of (CE) of -50 V and collision gas (CAD, nitrogen) of 5
psi.
[0353] Six of the eight metabolite masses previously discovered on
the FTMS were verified on the ABI Q-Star mass spectrometer. The
metabolites with the accurate masses of 723.5195 and 723.5197 were
determined to be the same metabolite, and the metabolite with
accurate mass of 886.5582 was not detected. Therefore, only six
metabolites (699.5198, 723.5195, 751.5555, 541.3432, 569.3687,
803.568) were used for the remaining analyses.
[0354] The extracted ion chromatograms (EICs) for the six
biomarkers are shown in FIG. 4 The top panel shows the eight
non-demented control EICs, and the bottom panel of each shows the
eight clinically-diagnosed AD EICs. The sensitivity of the Q-star
is superior to the FTMS, resulting in a greater magnitude in
intensity difference between the non-demented control subjects and
clinically diagnosed AD population for the selected biomarkers.
FIG. 5 shows the average raw intensity of the six biomarkers of the
eight non-demented control and eight clinically-diagnosed AD
samples as detected on the FTMS and Q-Star.
[0355] B. CSF Biomarkers
[0356] The metabolites and their associations with the clinical
variables described in this invention are further confirmed using
an independent mass spectrometry system. Representative sample
extracts from each variable group are re-analyzed by LC-MS using an
HP 1050 high-performance liquid chromatography, or equivalent
interfaced to an ABI Q-Star, or equivalent mass spectrometer to
obtain mass and intensity information for the purpose of
identifying metabolites that differ in intensity between the
clinical variables under investigation.
Example 3
Structure Elucidation of the Primary Metabolite Biomarkers
[0357] Characteristics that can be used for structure elucidation
of metabolites include accurate mass and molecular formula
determination, polarity, acid/base properties, NMR spectra, and
MS/MS or MSn spectra. These data, and in particular the MS/MS
spectrum, can be used as fingerprints of a particular metabolite
and are unique identifiers of a particular metabolite regardless of
whether the complete structure has been determined.
[0358] A. Serum Biomarkers--Structural Elucidation
[0359] 1. LC Retention Time.
[0360] The extracts containing the metabolites of interest were
subjected to reverse phase LC-MS using a C18 column and analysis by
MS as described in Example 2 above. Table 14 lists the resulting
retention times and detected masses for each of the six serum
metabolite markers. The retention time for all six of the
biomarkers is approximately 29-42 minutes under these HPLC
conditions.
[0361] 2. Extraction Conditions.
[0362] The conditions of extraction also provide insights about the
chemical properties of the biomarkers. All eight metabolites in the
serum (from Example 1) were ionized in negative mode (3 in APCI and
5 in ESI), which is indicative of a molecule containing an acidic
moiety such as a carboxylic acid or phosphate. Any moiety capable
of losing a hydrogen atom can be detected in negative ionization
mode. Three of the metabolite markers were extracted into an
organic ethyl acetate fraction (plasmalogen metabolites),
indicating that these metabolites are non-polar under acidic
condition; one was extracted into an organic ethyl acetate fraction
dried down and resuspended in butanol, indicating that this
metabolite (plasmalogen metabolite) is non-polar under acidic
conditions. Four of the metabolites (phosphatidyl choline related
metabolites) did not extract into the organic fraction, but rather
remained in the aqueous methanol/ammonium hydroxide fraction,
indicating that these metabolites are very polar.
[0363] 3. MS/MS Spectra.
[0364] The six serum metabolites identified as having the best
diagnostic ability were subject to MS/MS fragmentation using
collision induced dissociation (CID). The structure of a given
molecule will dictate a specific fragmentation pattern under
defined conditions that is specific for that molecule (equivalent
to a person's fingerprint). Even slight changes to the molecule's
structure can result in a different fragmentation pattern. In
addition to providing a fingerprint of the molecule's identity, the
fragments generated by CID can be used to gain insights about the
structure of a molecule, and for generating a very specific
high-throughput quantitative detection method (see [30-33] for
examples). FIGS. 6 through 11 show the MS/MS spectra for each of
the six markers at -50V collision energy (CE) voltages.
[0365] The masses resulting from CID MS/MS of each parent mass were
then used to calculate putative formulas for each of the fragment
ions for the metabolites specific to the ADAS-cog scores, as shown
in the tables for each marker (Tables 15 to 17). The information
inherent in the fragmentation data is highly specific and
descriptive for each metabolite, which can be used to gain
structural insights about each molecule. MS/MS was carried out on
the ABI-Q Star XL with all parameters as previously mentioned using
Nitrogen as the collision gas at 5 psi and collision energy (CE)
settings of -50 volts.
[0366] Based on the fragmentation pattern and masses, the
metabolite markers specific to the ADAS-cog scores have been
assigned structures having a phosphatidylcholine-related backbone.
From the CID MS/MS, the molecular formulae of 3 metabolites
specific to the ADAS-cog scores (accurate neutral masses of
541.3432, 569.3687, 803.568) were determined to be
C.sub.25H.sub.51NO.sub.9P, C.sub.27H.sub.55NO.sub.9P, and
C.sub.43H.sub.81NO.sub.10P, respectively. Their structures are
shown in FIGS. 12-14. The putative structures of additional markers
are shown in FIG. 15.
[0367] The 3 metabolites specific to AD pathology, with accurate
neutral masses of 751.5555, 699.5198, and 723.5195, were analyzed
using FT-ICRMS and LC/MS techniques, and by HRAPCI-MS, and MS/MS
spectral analysis. Daughter ions determined from the fragmentation
pattern for each metabolite marker are shown in FIGS. 16-18. The
molecular formulae were determined to be C.sub.43H.sub.78NO.sub.7P,
C.sub.39H.sub.74NO.sub.7P, and C.sub.41H.sub.74NO.sub.7P,
respectively. Based on the fragmentation pattern and masses, the
metabolite markers specific to AD pathology have been assigned
structures having an ethanolamine plasmalogen backbone.
[0368] For the 751.5555 metabolite (C.sub.43H.sub.78NO.sub.7P), and
due to negative ionization conditions, the HRAPCI-MS m/z measured
was 750.5482 ([M-H]-, calcd. 750.5477 for
C.sub.43H.sub.77NO.sub.7P). The relative intensity of the MS/MS
fragment masses (MS/MS m/z) were measured as follows: 750 ([M-H]-,
25%), 482 (1%), 464 (12%), 446 (5%), 329 (8%), 303 (100%), 259
(12%), 205 (8%), 140 (8%). The MS/MS fragments are shown in FIG.
16. The strong MS/MS fragment ion at m/z 303 and other fragment
ions due to loss of sn-2 acyl group (m/z 464) as a ketone, loss of
the sn-1 vinyl ether side chain (m/z 482) though small, and the
fragment ion due to phosphoethanolamine (m/z 140) indicated the
metabolite to be a plasmenyl phosphatidylethanolamine-type molecule
with arachidonic acid at the sn-2 position. Based on these results,
the structure of the 751.5555 metabolite was elucidated as
1-O-1'-(Z)-octadecenyl-2-arachidoyl-sn-glycero-3-phosphoethanolamine.
This was confirmed by comparison of their LC/MS and MS/MS spectral
data (FIG. 19)
[0369] The two remaining metabolites with molecular formulae
C.sub.39H.sub.74NO.sub.7P (neutral mass 699.5198) and
C.sub.41H.sub.74NO.sub.7P (neutral mass 723.5195) were found to
co-elute with the 751.5555 metabolite in LC/MS. The metabolites'
MS/MS fragment ions and fragmentation patterns were similar to
those of the 751.5555 metabolite.
[0370] For the 699.5198 metabolite, the HRAPCI-MS m/z measured was
698.5125 ([M-H]-, calcd. 698.5130 for C.sub.39H.sub.73NO.sub.7P).
The relative intensity of the MS/MS m/z were measured as follows:
698 ([M-H].sup.-, 8%), 536 (4%), 279 (100%), 255 (15%), 119 (10%).
The MS/MS fragments are shown in FIG. 17. Based on these results
and on its structural similarity to the 751.5555 metabolite, the
structure of the 699.5198 metabolite was determined to be
1-O-1'-(Z)-hexadecenyl-2-linoleyl-sn-glycero-3-phosphoethanolamine.
[0371] For the 723.5195 metabolite, the HRAPCI-MS m/z measured was
722.5124 ([M-H].sup.-, calcd. 722.5130 for
C.sub.41H.sub.73NO.sub.7P). The relative intensity of the MS/MS m/z
were measured as follows: 722 ([M-H].sup.-, 12%), 482 (1%), 436
(15%), 418 (6%), 303 (100%), 279 (6%), 259 (15%), 255 (10%), 205
(4%), 140 (5%). The MS/MS fragments are shown in FIG. 18. Based on
these results and on its structural similarity to the 751.5555
metabolite, the structure of the 723.5195 metabolite was proposed
as
1-0-1'-(Z)-hexadecenyl-2-arachidoyl-sn-glycero-3-phosphoethanolamine.
[0372] 4. NMR Spectra.
[0373] The MS/MS fragmentation provides highly specific descriptive
information about a metabolite. However, nuclear magnetic resonance
(NMR) can assist in solving and confirming the structures of the
metabolites. As NMR analysis techniques are typically less
sensitive than mass spectrometry techniques, multiple injections
are performed on the HPLC and the retention time window
corresponding to the metabolites of interest collected and
combined. The combined extract is then evaporated to dryness and
reconstituted in the appropriate solvent for NMR analysis.
[0374] Multiple NMR techniques and instruments are available, for
example, NMR spectral data are recorded on Bruker Avance 600 MHz
spectrometer with cryogenic probe after the chromatographic
separation and purification of the metabolites of interest. 1H NMR,
.sup.13C NMR, noe-difference spec, as well as 2-D NMR techniques
like heteronuclear multiple quantum correlation (HMQC), and
heteronuclear multiple bond correlation (HMBC) are used for
structure elucidation work on the biomarkers.
[0375] B. CSF Biomarkers
[0376] The structural characteristics (LC retention time,
extraction conditions, MS/MS fragments) for the 12 CSF metabolite
markers are determined in the same manner as detailed above.
Example 4
Characterization of Ethanolamine Phospholipids in Serum
[0377] Based on the fact that the metabolite markers specific to AD
pathology have an ethanolamine plasmalogen backbone, it was further
investigated whether other serum plasmalogens could be indicative
of AD. This characterization of ethanolamine phospholipids in serum
was made using a chromatographic method combined with a mass
spectrometric detector.
[0378] For MS/MS applications and experiments involving
chromatography, an Agilent 1100 HPLC system was used in combination
with an Applied Biosystems QSTAR XL mass spectrometer. An Agilent
Zorbax RX-SIL (4.6.times.150 mm, 5 .mu.m) column was used for
normal phase chromatography. Conditions included an isocratic
mobile phase (55:40:5 isopropanol:hexane:H2O) at a flow rate of 1.0
mL/min for a total run time of 15 min. The column was heated to
35.degree. C. The sample injection volume was 10 .mu.L. Organic
solvent extracts (ethyl acetate) of samples were evaporated to
dryness under nitrogen gas and the residue was reconstituted in 100
.mu.L of 55:40:5 isopropanol:hexane:H2O solution prior to
injection.
[0379] The QSTAR XL instrument was equipped with an APCI (Heated
Nebulizer) source operating in negative mode. Values of major
instrument parameters were DP, -60; FP, -265; DP2, -15; GS1, 75;
GS2, 15; CUR, 30; NC, -3; TEM, 400.degree. C.; Scan range,
50-1500amu; Accumulation time, I sec.
[0380] The three classes of ethanolamine phospholipids are
described as diacyl (also referred to herein as PtdEt), alkyl-acyl
(also referred to herein as plasmanyl) or alkenyl-acyl (also
referred to herein as EtnPl or plasmenyl). Various basic structures
of ethanolamine phospholipids are shown in FIG. 20, along with the
standard naming convention used herein. Table 18 shows a list of
plasmanyl and plasmenyl ethanolamine phospholipids (M5-M24) that
are presently identified and are of particular interest.
[0381] FIGS. 21-32 show structural information pertaining to
selected metabolites detected in serum. These figures illustrate
the retention time, MS/MS fragmentation patterns, and putative
structures for selected molecules. Due to the conserved MS/MS
fragmentation mechanism between these molecules, the theoretical
MS/MS transition can be determined for any ethanolamine
phospholipid by using a combination of the parent ion mass and the
fragment mass of the moiety at either the sn-1 or sn-2
position.
Example 5
High Throughput Commercial Method Development
[0382] A high throughput method for differential diagnosis of AD
dementia and non-AD dementia states was established.
[0383] High throughput screening (HTS) was performed with a linear
ion trap mass spectrometer (Q-trap 4000, Applied Biosystem) coupled
with Agilent 1100 LC system. Sample was prepared by adding 15 uL of
internal standard (5 .mu.g/mL of (24-13C)-Cholic Acid in methanol)
to 120 uL ethyl acetate fraction of each sample. 100 ul sample was
injected by flow injection analysis (FIA), and monitored under
negative APCI mode. The method was based on multiple reaction
monitoring (MRM) scan mode of one parent/daughter transition for
each metabolite and one internal standard. Each transition was
scanned for 70 ms for a total cycle time of 2.475 sec. The
isocratic 10% EtOAc in MeOH elution was performed with a flow rate
at 360 .mu.l/min for 1 min. The source parameters were set as
follows: CUR: 10.0, CAD: 8, NC: -4.0, TEM: 400, GS1: 30, GS2: 50,
interface heater on. The compound parameters were set as follows:
DP: -120.0, EP: -10, NC: -4.0, CE: -40, CXP: -15. FIG. 33
illustrates a representative standard curve for this method for
EtnPls 16:0/22:6 generated by diluting a normal serum sample while
maintaining a constant concentration of internal standard
(24-13C)-Cholic Acid).
Example 6
Effect of Aging and Severity of Dementia on Serum Levels of
Ethanolamine Phospholipids
[0384] The effect of aging and severity of dementia on serum levels
of ethanolamine phospholipids in 752 subjects aged 40-95 with
various levels of dementia was investigated. The clinical data on
the subject cohorts is shown in Table 19.
[0385] The effect of age was evaluated using a set of aged 30 to 95
subjects of untested cognitive status who did not suffer from
dementia. Subjects were divided into one of five subgroups based
upon their decade of life (30's, 40's, 50's, 60's, and >70). The
40-49 cohort was used as the pre-dementia reference group due to
the low incidence of dementia at this age. The metabolites of
interest (see Table 18) were measured using the high throughput
method described in Example 5.
[0386] The effect of dementia severity was determined in subjects
aged 56 to 95, comprised of 68 cognitively confirmed non-demented
subjects (MMSE.gtoreq.28); 256 subjects currently diagnosed with
SDAT (ADAS-cog 6-70, MMSE 0-26); 20 post-mortem confirmed SDAT and
20 post-mortem confirmed controls. Subjects were grouped into one
of four dementia subgroups based upon either their MMSE score
[.gtoreq.28=Cognitively Normal] or their ADAS-cog score [5-19 =low
cognitive impairment); 20-39 =moderate; 40-70 =severe].
6A. Absolute Levels of Ethanolamine Phospholipids
[0387] A significant gender bias was observed in that only females
exhibited an age-related decrease in EtnPls. Free docosohexanoic
acid (DHA, Free 22:6, M25) in both males and females was
significantly increased in the 50-59, 60-69, and 70+cohorts
relative to the 40-49 cohorts. However, only males exhibited a
concomitant increase in both 16:0/22:6-EtnPl (M19) and
18:0/22:6-EtnPl (M24) (see Tables 20-21 for males; Tables 22-23 for
females). These data indicate that, in females, there may be an
age-related dysfunction in the packaging of DHA into EtnPls. This
gender difference may explain the increased incidence of dementia
in very old females (19).
[0388] In both males and females, the majority of EtnPls in all
dementia subgroups were significantly reduced relative to cognitive
controls. In both males and females, free DHA (M25) was
significantly decreased only in severely demented subjects. In
females, a dementia effect was observed for three EtnPls (16:0/18:2
(M16), 18:0/18:2 (M21), and 16:0/20:4 (M17)) in that both
18:2-containing EtnPls were significantly lower in severely
demented subjects versus either low or moderately demented females,
and 16:0/20:4 (M17) was lower in the severe group versus the low
group (see Tables 24-26). In males, a dementia effect was observed
for DHA (M25) and 16:0/22:6 (M19) in that free DHA (M25) was
reduced in the moderate group versus the low group and in the
severe group versus the moderate group and 16:0/22:6 (M19) was
reduced in the severe group versus the low group (see Tables
27-29). These results indicate that the progressive cognitive
deterioration in AD manifests slightly differently in the two
sexes.
[0389] Brain white matter contains primarily 18:1- and
18:2-containing EtnPls with low levels of 20:4-containing and
22:6-containing EtnPls, whereas gray matter contains significantly
higher levels of 20:4-containing and 22:6-containing EtnPls [34].
In females, increasing dementia appears to affect both white (18:2)
and gray (20:4) matter EtnPls equally, whereas in males
predominantly gray (22:6) matter EtnPls appear to be affected to a
greater extent.
[0390] Post-mortem collected serum samples from 20 pathologically
confirmed AD subjects and 20 subjects containing minimal amyloid
deposition were also analyzed. Both gray and white matter EtnPls
were significantly decreased in post-mortem confirmed AD relative
to age matched controls (see Tables 30 and 31).
6B. Relative Levels of Ethanolamine Phospholipids
[0391] The data collected above was re-analyzed to obtain a ratio
between the levels of each ethanolamine phospholipids with
16:0/18:0 PtdEt (M01). Measurement of the ethanolamine phospholipid
levels in this manner is more stable and reproducible than
measurement of the absolute levels. Furthermore, because the
16:0/18:0 PtdEt (M01) is naturally present in all samples and does
not appear to vary significantly over disease states, this approach
minimizes the sample-to-sample variability (due to handling,
extraction, etc).
[0392] The results obtained further support the observations and
conclusions made in 6A. The gender bias was with respect to an
age-related decrease in EtnPls was evident in data where ratios to
M01 were measured (see Tables 32-33 for males; Tables 34-35 for
females). The same trends with respect to the severity of cognitive
impairment were also observed (see Tables 36-38 for males; Tables
39-41 for females). In addition, pathology results on post-mortem
serum samples show similar trends (Tables 42 and 43).
[0393] Both the absolute EtnPls levels and the EtnPls to M01 ratio
exhibited a significant dementia effect. The EtnPls to M01 ratios
of all eight EtnPls (16:0/18:1 (M15), 16:0/18:2 (M16), 16:0/20:4
(M17), 16:0/22:6 (M19), 18:0/18:1 (M20), 18:0/18:2 (M21), 18:0/20:4
(M22), 18:0/22:6 (M24)) were significantly lower in the severely
demented group relative to the low group while six of the eight
were significantly lower in the severe group relative to the
moderate group
Example 7
The Grey and White Matter Score Distribution
[0394] A white and gray matter specific EtnPl scoring system was
developed whereby each EtnPl in each subject was normalized to
their respective gender-specific cognitively normal mean, log2
transformed and mean centered. Each subject's white matter score
was taken as the lowest such value of plasmenyl 16:0/18:1 (M15),
16:0/18:2 (M16), 18:0/18:1 (M20), and 18:0/18:2 (M21) EtnPls, and
their gray matter score as the lowest of plasmenyl 16:0/20:4 (M17),
16:0/22:6 (M19), 18:0/20:4 (M22), and 18:0/22:6 (M24) EtnPls.
[0395] These simplified scores revealed that both gray and white
matter EtnPls were decreased at all stages of AD (FIG. 39) and that
the levels in post-mortem confirmed AD closely matched levels in
severely demented subjects of both sexes (Tables 44-45). The
cross-sectional white and gray matter score distributions in
subjects of various levels of dementia clearly showed a dementia
dependent shift in the population means (Tables 46-47). The effect
of age on the white and gray matter scores was also determined
(Tables 48-49). This also indicated that changes in serum levels of
gray matter EtnPls may precede white matter changes and potentially
be an early risk factor for AD. Such cross-sectional data does not
account for baseline variability among subjects. Individual
longitudinal trajectories of these scores may be more accurate at
detecting early risk of AD in otherwise healthy, non-demented
subjects.
[0396] Based on these scores, risk prediction can be performed on
both male and female subjects (Tables 49-50) where a cut-off value
that results in approximately 20-30% of cognitively normal subjects
being classified as either intermediate or high risk is used. Using
this cut-off value, a subject's white and gray matter score is
evaluated. If the subject tests normal on both scores, the subject
is deemed to be at low risk. If the subject tests positive on one
of the scores, the subjects is deemed to be at intermediate risk
and if the subject tests positive on both scores, the subject is
deemed to be at high risk.
Example 8
Effect of Dementia Severity and AD Pathology on Serum EtnPls Levels
in Combined Male and Female Subjects
[0397] The effect of dementia severity was determined using 324
subjects (176 female, 148 male) aged 56 to 95, comprised of 68
cognitively confirmed non-demented subjects (MMSE.gtoreq.28) and
256 subjects currently diagnosed with AD (ADAS-cog 6-70, MMSE
0-26). The effect of AD pathology was determined using serum
samples collected from 20 post-mortem confirmed AD and 19 control
subjects (Table 19). Subjects were grouped into one of four
dementia severity cohorts based upon either their MMSE score
[.gtoreq.28 =Cognitively Normal] or their ADAS-cog score [5-19 =low
cognitive impairment; 20-39 =moderate; 40-70 =severe].
[0398] Mean serum levels of 16:0/18:1 (M15), 16:0/18:2 (M16),
16:0/20:4 (M17), 16:0/22:6 (M19), 18:0/18:1 (M20), 18:0/18:2 (M21),
18:0/20:4 (M22), 18:0/22:6 (M24) EtnPls; free docosahexaenoic acid
(DHA, Free 22:6, M25); and phosphatidylethanolamine (PtdEt)
16:0/18:0 (D16:0/18:0; M01) were determined for each group (FIG.
34). All eight EtnPls in all dementia subgroups were observed to be
significantly reduced relative to cognitive controls (24 pair-wise
comparisons, t-test p-values 2.6e-2 to 2.0e-10, median =3.9e-5).
Free DHA (M25) was significantly decreased in both moderately and
severely demented subjects (p<0.05). All eight EtnPls were also
significantly decreased in post-mortem confirmed SDAT relative to
age matched controls. D16:0/18:0 (M01) levels, a non-plasmalogen
phoshopholipid remained unchanged across the different dementia
cohorts.
Example 9
Population Distributions as a Function of Dementia Severity
[0399] The EtnPls 16:0/22:6 (M19) to PtdEt 16:0/18:0 ratio (M01)
(DHA-EtnPls) showed the strongest overall sex-independent dementia
effect (Tables 38, 41) and was used for all subsequent population
distributions and comparisons. A summary of the key comparisons
using this ratio are listed in Table 52. This ratio was then log(2)
transformed and used to create a population histogram for each
cohort of increasing cognitive impairment (FIG. 35). A cut-off
value was selected based upon the findings of Bennett et al [35],
(i.e. .about.30% of the CN group being detected as AD) (FIG. 35,
dotted line). Using this cut-off, 63%, 79% and 83% of low, moderate
and severely demented subjects, respectively, were subsequently
classified as AD.
[0400] To compare these distributions with the known distributions
of A.beta. pathology in AD, the results of four prospective
pathology studies [8, 35-37] were combined to generate the
theoretical population distributions of A.beta. pathology in
demented and non-demented populations, assuming that A.beta. is
normally distributed in each population (FIG. 36A). These studies
reported that only 71% (140/198) of clinically diagnosed AD
subjects have AD pathology at death and that 32% (87/268) of
cognitively normal subjects meet neuropathological criteria for AD
at death. When the data from all of our cognitively tested subjects
were combined, 32% (22/68) of our non-demented population and 75%
(192/256) of our demented population were classified as AD positive
based upon their serum EtnPls level (FIG. 36B). This comparison
revealed that the observed distribution of depleted 22:6-containing
EtnPls perfectly matched the theoretical distribution of AD
pathology in demented and non-demented subjects.
Example 10
Linear Extrapolation of Disease Progression and Serum EtnPls
Depletion
[0401] To investigate whether a correlation between the decrease in
EtnPls and increasing dementia in the clinically diagnosed AD
population exists, a linear regression analysis was performed using
the mean 22:6-containing EtnPls level (normalized to CN) of each of
the dementia cohorts and the average ADAS-cog score for each of
these three cohorts (FIG. 37). A very high correlation was observed
between the mean 22:6-containing EtnPls level and the mean ADAS-cog
scores of the three dementia cohorts (r2=0.99). However, this
linear decrease did not extrapolate back to the CN group (X vs.
CN). Assuming a clinical AD progression of 7.5 ADAS-cog units per
year, this extrapolation predicts that that 22:6-containing EtnPls
levels begin to decline approximately seven years before clinical
cognitive impairment (ADAS-cog =15) is evident.
Example 11
The Effect of Chronological Age on Serum DHA-EtnPls Levels
[0402] To investigate whether the above prediction could be
verified experimentally, the serum 22:6-containing EtnPls levels in
209 subjects (110 male, 99 female, Table 19) of unknown cognitive
status but currently not diagnosed with dementia was determined and
compared to the clinical AD and CN cohorts (FIG. 38). The results
of this analysis revealed a significant drop in serum
22:6-containing EtnPls in the aged 60-69 cohort versus the aged
50-59 cohort (FIG. 38A). This cohort also had significantly lower
levels versus the CN group even though the CN group was, on
average, 13 years older. Interestingly, the aged 70-95 cohort was
not significantly different from either the aged 50-59 cohort or
the CN cohort, but had significantly higher levels than the SDAT
cohort.
Example 12
Sub-Populations Identified by Serum DHA-EtnPls Levels
[0403] The distribution of serum DHA-EtnPls within each age group,
as shown in FIG. 38B, was also examined. The population
distributions of the five groups (three age groups, CN and AD)
differentiated by age and dementia status reveal the presence of
three distinct populations (P1-P3, FIG. 38B). The populations were
assigned as: P1--subjects with AD pathology and no remaining
reserve capacity; P3--subjects with little or no AD pathology;
P2--subjects that are transitioning from P3 to P1. These P2
subjects are hypothesized to have AD pathology and some level of
reserve remaining.
[0404] Since AD subjects have a life expectancy of less than 10
years from diagnosis [38, 39] and low 22:6-containing EtnPls are
highly associated with AD severity, the decreased number of P1
subjects observed in the aged 70-95 cohort is most likely due to
differences in life expectancy between P1 and P2 or P3. The
transitory nature of P2 is best illustrated by examining the
different ratios between the percentages of subjects present in P3
compared to P2, as observed in the lower three panels of FIG. 7B.
These three cohorts differ only in dementia status. The P3 to P2
ratio changes from 3:1 (68% versus 22%) in the confirmed cognitive
normal group to an intermediate ratio of 1:1 (43% versus 46%) in
the normal healthy elderly group of unknown cognitive status, to
0.6:1 (25% versus 40%) in the confirmed demented AD cohort.
[0405] All citations are hereby incorporated by reference.
[0406] The present invention has been described with regard to one
or more embodiments. However, it will be apparent to persons
skilled in the art that a number of variations and modifications
can be made without departing from the scope of the invention as
defined in the claims.
TABLE-US-00001 TABLE 1 Accurate mass features differing between
clinically diagnosed AD patients and non-demented controls (p <
0.05, log2 transformed). Detected Analysis AVG (log2) SEM AVG
(log2) SEM log(2) Mass Mode AD AD Normal Normal Ratio P Value
723.5197 1204 3.576 0.039 4.350 0.056 0.822 5.09E-19 723.5195 1202
2.186 0.040 2.892 0.052 0.756 4.86E-17 724.5257 1204 2.866 0.032
3.463 0.055 0.828 4.15E-16 749.5367 1202 3.176 0.034 3.714 0.041
0.855 3.82E-15 751.5555 1204 4.575 0.041 5.248 0.060 0.872 1.07E-14
751.5529 1202 3.335 0.036 3.920 0.050 0.851 1.14E-14 752.5564 1202
2.251 0.038 2.836 0.050 0.794 3.13E-14 752.5583 1204 3.472 0.042
4.094 0.061 0.848 6.47E-13 699.5198 1204 2.216 0.038 2.775 0.054
0.799 6.76E-13 750.544 1204 3.279 0.038 3.858 0.063 0.850 1.07E-12
749.5407 1204 4.426 0.039 5.012 0.064 0.883 1.33E-12 541.3432 1102
3.315 0.033 3.798 0.048 0.873 1.42E-12 750.5402 1202 2.192 0.038
2.704 0.044 0.811 2.76E-12 725.5385 1204 2.884 0.043 3.417 0.054
0.844 1.28E-10 569.3687 1102 2.262 0.039 2.724 0.048 0.830 6.22E-10
727.5568 1204 3.518 0.038 3.986 0.060 0.882 1.70E-09 804.5713 1102
4.207 0.042 4.610 0.033 0.913 3.88E-08 803.568 1102 5.432 0.043
5.838 0.035 0.930 7.10E-08 726.5461 1204 2.808 0.032 3.150 0.050
0.892 7.50E-08 827.57 1102 4.151 0.048 4.630 0.062 0.897 9.42E-08
803.5445 1101 5.123 0.055 5.655 0.059 0.906 1.01E-07 555.3102 1102
1.818 0.045 2.240 0.046 0.812 1.27E-07 565.3394 1102 3.480 0.050
3.958 0.055 0.879 1.28E-07 804.5476 1101 4.169 0.056 4.703 0.060
0.887 1.33E-07 828.5737 1102 3.138 0.046 3.590 0.061 0.874 1.89E-07
567.3547 1102 2.822 0.041 3.218 0.054 0.877 2.89E-07 728.5627 1204
2.935 0.033 3.281 0.060 0.895 5.16E-07 817.5377 1102 2.282 0.048
2.712 0.057 0.842 8.85E-07 779.5444 1101 6.433 0.053 6.874 0.043
0.936 1.06E-06 780.5474 1101 5.437 0.053 5.875 0.043 0.925 1.28E-06
812.5762 1202 1.659 0.050 2.084 0.058 0.796 2.24E-06 832.6026 1102
3.455 0.041 3.795 0.040 0.910 2.48E-06 811.5732 1202 2.705 0.036
3.027 0.055 0.893 3.40E-06 871.5528 1102 3.068 0.042 3.408 0.040
0.900 3.47E-06 831.5997 1102 4.564 0.042 4.903 0.040 0.931 3.48E-06
793.5386 1102 3.604 0.043 3.950 0.039 0.912 3.50E-06 782.5085 1204
3.401 0.045 3.780 0.055 0.900 3.81E-06 805.5832 1102 4.075 0.047
4.485 0.068 0.909 3.87E-06 781.5617 1101 6.109 0.061 6.610 0.072
0.924 5.14E-06 813.5885 1202 3.012 0.030 3.276 0.048 0.919 6.23E-06
794.5421 1102 2.523 0.042 2.853 0.040 0.885 6.30E-06 814.5917 1202
2.041 0.026 2.289 0.051 0.892 7.46E-06 747.5245 1204 3.473 0.043
3.886 0.090 0.894 9.55E-06 837.5027 1101 3.578 0.045 3.933 0.050
0.910 1.02E-05 782.565 1101 5.083 0.063 5.589 0.078 0.909 1.09E-05
746.5717 1204 3.085 0.031 3.362 0.061 0.918 1.82E-05 829.5856 1102
4.043 0.048 4.398 0.046 0.919 1.85E-05 784.5237 1204 3.310 0.040
3.603 0.037 0.919 1.91E-05 786.5416 1204 3.815 0.035 4.087 0.043
0.933 1.91E-05 760.5216 1204 4.075 0.036 4.347 0.039 0.938 2.11E-05
745.5658 1204 3.937 0.034 4.242 0.068 0.928 2.12E-05 744.5536 1204
4.322 0.034 4.605 0.058 0.939 2.46E-05 783.5672 1101 3.755 0.068
4.259 0.079 0.882 3.48E-05 807.5758 1101 5.736 0.052 6.102 0.047
0.940 3.69E-05 808.5792 1101 4.697 0.052 5.063 0.047 0.928 4.20E-05
743.5471 1204 5.286 0.036 5.579 0.063 0.947 4.94E-05 482.3215 1202
1.971 0.038 2.251 0.062 0.875 0.0001 755.486 1204 3.221 0.047 3.561
0.057 0.905 0.0001 758.5092 1204 4.574 0.033 4.808 0.042 0.951
0.0001 775.5533 1202 2.120 0.044 2.449 0.068 0.866 0.0001 787.5729
1202 1.847 0.040 2.145 0.057 0.861 0.0001 795.5181 1101 2.630 0.059
3.044 0.064 0.864 0.0001 795.555 1102 2.665 0.043 2.986 0.065 0.892
0.0001 805.5605 1101 5.414 0.057 5.785 0.048 0.936 0.0001 831.5759
1101 4.297 0.056 4.677 0.060 0.919 0.0001 855.6016 1102 3.538 0.045
3.873 0.065 0.914 0.0001 517.314 1101 5.470 0.038 5.755 0.069 0.951
0.0002 541.3139 1101 4.091 0.053 4.494 0.096 0.910 0.0002 542.3173
1101 2.284 0.055 2.687 0.089 0.850 0.0002 747.5201 1202 1.937 0.051
2.313 0.088 0.838 0.0002 757.4991 1101 3.644 0.065 4.073 0.070
0.895 0.0002 775.5528 1204 3.197 0.045 3.537 0.085 0.904 0.0002
806.5639 1101 4.423 0.057 4.779 0.049 0.926 0.0002 832.5791 1101
3.357 0.055 3.723 0.061 0.902 0.0002 915.5191 1101 2.376 0.051
2.717 0.063 0.874 0.0002 755.5468 1101 2.326 0.064 2.753 0.085
0.845 0.0003 777.553 1202 1.859 0.072 2.384 0.130 0.780 0.0003
829.5604 1101 3.535 0.053 3.875 0.058 0.912 0.0003 518.3174 1101
3.438 0.038 3.717 0.073 0.925 0.0004 731.5464 1101 1.945 0.096
2.600 0.157 0.748 0.0004 757.5626 1101 6.655 0.074 7.108 0.079
0.936 0.0004 758.5656 1101 5.702 0.075 6.160 0.078 0.926 0.0004
759.5779 1101 5.547 0.069 5.997 0.099 0.925 0.0004 760.5811 1101
4.419 0.071 4.886 0.103 0.904 0.0004 768.5539 1204 3.964 0.046
4.279 0.077 0.927 0.0004 748.5287 1204 2.277 0.091 2.820 0.088
0.807 0.0005 783.5148 1204 3.284 0.047 3.574 0.056 0.919 0.0005
821.5712 1102 3.064 0.040 3.304 0.044 0.927 0.0005 523.4679 1203
3.252 0.121 4.011 0.165 0.811 0.0006 781.562 1201 7.364 0.040 7.638
0.071 0.964 0.0006 810.5399 1204 2.835 0.050 3.145 0.066 0.901
0.0006 732.4938 1204 4.249 0.042 4.511 0.057 0.942 0.0007 522.4635
1203 4.713 0.126 5.485 0.176 0.859 0.0008 810.5969 1101 3.957 0.072
4.370 0.080 0.905 0.0009 853.5854 1102 2.626 0.043 2.880 0.052
0.912 0.0009 819.5551 1102 2.394 0.046 2.654 0.046 0.902 0.001
828.5743 1202 5.243 0.058 5.598 0.085 0.936 0.001 478.254 1201
1.584 0.083 1.051 0.148 1.507 0.0011 579.5325 1203 4.102 0.089
4.636 0.127 0.885 0.0011 744.4956 1204 3.881 0.039 4.117 0.058
0.943 0.0011 761.5843 1101 2.422 0.065 2.829 0.106 0.856 0.0011
809.5936 1101 4.964 0.070 5.362 0.079 0.926 0.0011 886.5582 1102
3.041 0.037 3.250 0.041 0.936 0.0011 481.3172 1202 3.923 0.039
4.159 0.060 0.943 0.0012 767.5495 1204 5.063 0.049 5.369 0.082
0.943 0.0012 782.5653 1201 6.339 0.043 6.614 0.076 0.959 0.0012
827.5701 1202 6.306 0.063 6.684 0.092 0.943 0.0012 847.5316 1101
2.742 0.059 3.091 0.083 0.887 0.0012 789.5892 1202 1.723 0.036
1.952 0.065 0.883 0.0013 543.3296 1101 3.733 0.044 4.023 0.087
0.928 0.0014 575.2728 1101 2.470 0.043 2.739 0.074 0.902 0.0014
580.5351 1203 1.456 0.127 2.192 0.174 0.664 0.0014 521.4522 1203
2.302 0.131 3.028 0.158 0.760 0.0016 731.4916 1204 5.482 0.046
5.746 0.063 0.954 0.0016 759.5163 1204 4.724 0.042 4.954 0.048
0.954 0.0016 306.2569 1204 2.998 0.046 3.266 0.067 0.918 0.0017
771.5814 1204 4.164 0.036 4.367 0.046 0.954 0.0017 786.5967 1101
4.808 0.074 5.203 0.078 0.924 0.0019 458.2405 1101 1.736 0.040
1.986 0.077 0.874 0.0021 520.4499 1203 3.956 0.112 4.577 0.148
0.864 0.0021 748.5735 1202 3.918 0.035 3.722 0.050 1.053 0.0021
490.3641 1203 1.944 0.092 1.397 0.159 1.391 0.0023 545.3453 1101
3.606 0.051 3.898 0.078 0.925 0.0023 605.5457 1203 5.135 0.068
5.509 0.089 0.932 0.0023 769.5656 1204 3.963 0.038 4.174 0.054
0.950 0.0023 570.3725 1202 2.976 0.032 3.155 0.045 0.943 0.0024
785.5933 1101 5.884 0.074 6.271 0.080 0.938 0.0024 582.2473 1201
3.325 0.096 2.793 0.141 1.191 0.0026 569.369 1202 4.908 0.033 5.089
0.047 0.964 0.0027 784.5811 1101 4.405 0.079 4.817 0.089 0.915
0.0027 811.6096 1101 3.078 0.083 3.511 0.096 0.877 0.0027 590.343
1202 4.025 0.050 4.304 0.076 0.935 0.0028 856.672 1202 2.764 0.038
2.553 0.057 1.082 0.0028 833.5932 1101 3.276 0.068 3.608 0.056
0.908 0.003 506.2851 1201 3.142 0.077 2.656 0.165 1.183 0.0031
793.5681 1204 3.155 0.040 3.372 0.055 0.936 0.0031 546.3485 1101
1.999 0.050 2.287 0.089 0.874 0.0036 591.3542 1202 4.045 0.061
4.345 0.058 0.931 0.0037 741.5305 1204 2.931 0.056 3.250 0.102
0.902 0.0042 796.5876 1204 2.634 0.043 2.860 0.062 0.921 0.0042
804.7227 1203 1.842 0.145 2.611 0.220 0.705 0.0044 807.59 1202
2.463 0.045 2.718 0.082 0.906 0.0045 506.3213 1202 2.538 0.040
2.748 0.061 0.923 0.0046 552.5022 1203 3.164 0.088 3.643 0.147
0.869 0.0047 589.3403 1202 5.876 0.056 6.171 0.085 0.952 0.0048
806.5873 1202 4.366 0.047 4.635 0.092 0.942 0.0048 550.4957 1203
6.898 0.096 7.415 0.160 0.930 0.0049 604.5433 1203 6.554 0.069
6.901 0.089 0.950 0.005 805.5839 1202 5.562 0.048 5.841 0.097 0.952
0.0052 551.4986 1203 5.480 0.095 5.988 0.158 0.915 0.0053 743.5469
1202 3.061 0.055 3.348 0.086 0.914 0.0056 541.3435 1202 5.669 0.059
5.974 0.094 0.949 0.0058 183.0661 1101 2.590 0.091 3.015 0.094
0.859 0.006 858.6212 1202 2.707 0.059 2.994 0.074 0.904 0.0061
614.4914 1203 2.779 0.060 2.448 0.114 1.135 0.0062 787.5465 1204
2.675 0.077 3.017 0.054 0.887 0.0062 772.5862 1204 3.287 0.031
3.437 0.042 0.956 0.007 837.5881 1202 2.429 0.027 2.577 0.053 0.942
0.007 509.3493 1202 2.403 0.035 2.579 0.055 0.931 0.0071 529.3167
1202 3.032 0.048 3.265 0.069 0.928 0.0075 564.5134 1203 2.706 0.075
3.075 0.111 0.880 0.0075 566.3434 1202 5.203 0.049 5.436 0.062
0.957 0.0075 833.7571 1203 2.962 0.109 3.507 0.170 0.845 0.0077
631.628 1203 1.795 0.127 2.391 0.161 0.751 0.008 857.6186 1202
3.773 0.058 4.049 0.076 0.932 0.008 858.6861 1202 2.943 0.040 2.756
0.052 1.068 0.0081 519.3321 1101 3.964 0.084 4.382 0.133 0.905
0.0083 685.26 1202 1.771 0.048 1.998 0.063 0.886 0.0083 757.5014
1204 3.755 0.048 3.971 0.052 0.946 0.0085 744.55 1202 1.968 0.051
2.222 0.083 0.886 0.0086 671.5723 1204 2.231 0.084 2.604 0.087
0.857 0.0087 304.241 1204 4.887 0.041 5.088 0.066 0.961 0.0092
536.4794 1203 2.320 0.101 2.799 0.143 0.829 0.0093 542.3461 1202
3.873 0.049 4.106 0.074 0.943 0.0095 675.6377 1204 3.953 0.043
4.160 0.066 0.950 0.0098 520.3354 1101 2.240 0.085 2.646 0.130
0.846 0.01 832.7523 1203 3.859 0.107 4.374 0.169 0.882 0.0103
409.0208 1202 2.806 0.035 2.980 0.063 0.942 0.0106 768.5503 1202
1.942 0.076 2.277 0.090 0.853 0.0111 303.1079 1202 5.648 0.034
5.802 0.047 0.973 0.0113 592.3571 1202 2.291 0.063 2.560 0.062
0.895 0.0115 837.718 1204 2.888 0.176 3.662 0.217 0.788 0.0121
832.7492 1204 3.286 0.119 3.848 0.190 0.854 0.0125 832.6037 1202
5.066 0.047 5.274 0.062 0.961 0.013 411.3212 1202 2.868 0.038 3.033
0.048 0.946 0.0134 838.7226 1204 2.313 0.148 2.973 0.206 0.778
0.0136 670.569 1204 3.239 0.061 3.515 0.093 0.921 0.0141 795.5838
1204 3.566 0.046 3.769 0.063 0.946 0.0141 767.547 1202 3.073 0.064
3.358 0.092 0.915 0.0143 305.2438 1204 2.519 0.044 2.719 0.067
0.926 0.0146 505.3229 1202 3.994 0.051 4.222 0.076 0.946 0.0156
803.5677 1202 7.196 0.070 7.502 0.098 0.959 0.0157 711.2577 1202
2.250 0.049 2.454 0.056 0.917 0.0159 827.5448 1101 3.549 0.077
3.873 0.095 0.916 0.016 548.4815 1203 7.094 0.072 7.405 0.103 0.958
0.0174 568.3573 1202 4.008 0.035 4.167 0.060 0.962 0.0175 578.5277
1203 4.301 0.303 5.548 0.355 0.775 0.0175 601.5164 1203 7.640 0.038
7.463 0.069 1.024 0.0185 549.4845 1203 5.666 0.077 5.994 0.110
0.945 0.0187 743.5466 1203 1.987 0.064 2.266 0.099 0.877 0.0189
772.5278 1204 3.324 0.039 3.488 0.055 0.953 0.019 765.5334 1204
3.269 0.060 3.540 0.104 0.923 0.0193 440.3532 1204 1.417 0.096
0.975 0.180 1.453 0.0205 495.332 1101 5.251 0.073 5.565 0.112 0.944
0.0205 804.5718 1202 5.877 0.057 6.117 0.080 0.961 0.0206 340.2976
1203 1.597 0.082 1.937 0.112 0.825 0.0208 856.6061 1202 4.565 0.056
4.805 0.086 0.950 0.0212 584.2646 1204 3.136 0.115 2.649 0.172
1.184 0.0218 733.6426 1204 2.978 0.049 2.718 0.126 1.096 0.0219
588.4731 1203 2.387 0.064 2.031 0.179 1.175 0.0223 765.5313 1202
1.802 0.073 2.098 0.095 0.859 0.0226 523.3634 1101 3.466 0.082
3.791 0.103 0.914 0.0235 830.5894 1202 4.847 0.051 5.051 0.064
0.960 0.0236 887.7352 1204 6.295 0.082 5.963 0.113 1.056 0.0244
598.5124 1204 2.094 0.134 1.513 0.233 1.385 0.0249 616.5052 1203
4.205 0.062 3.957 0.084 1.062 0.025 916.7743 1204 5.894 0.081 5.571
0.107 1.058 0.025 430.3818 1204 4.938 0.075 5.253 0.122 0.940
0.0262 855.6023 1202 5.552 0.056 5.784 0.087 0.960 0.0266 684.5489
1204 2.523 0.055 2.745 0.080 0.919 0.0269 831.6001 1202 6.192 0.053
6.404 0.074 0.967 0.0269 826.7069 1204 2.482 0.089 2.819 0.099
0.880 0.0275 915.7681 1204 6.304 0.085 5.972 0.116 1.056 0.0293
615.3539 1202 2.463 0.043 2.629 0.058 0.937 0.0296 431.386 1204
3.491 0.075 3.795 0.118 0.920 0.0298 942.7879 1204 3.487 0.078
3.195 0.092 1.091 0.0302 665.501 1204 3.388 0.071 3.074 0.142 1.102
0.0305 861.7806 1203 2.531 0.099 2.902 0.119 0.872 0.0306 618.4829
1201 1.791 0.120 1.310 0.185 1.367 0.0309 830.7352 1204 4.252 0.096
4.638 0.150 0.917 0.031 801.555 1202 2.660 0.056 2.881 0.082 0.923
0.0311 739.5143 1204 2.754 0.087 3.121 0.158 0.883 0.0317 492.3816
1203 3.513 0.055 3.302 0.077 1.064 0.0339 741.5319 1202 1.106 0.080
1.424 0.125 0.777 0.0339 914.7583 1204 5.082 0.083 4.763 0.118
1.067 0.0339 507.3316 1202 2.940 0.035 3.094 0.071 0.950 0.034
504.3814 1203 1.701 0.070 1.413 0.124 1.204 0.0341 496.3355 1101
3.442 0.074 3.736 0.119 0.921 0.0347 521.3477 1101 3.717 0.075
4.021 0.127 0.925 0.0351 829.5859 1202 5.976 0.056 6.181 0.070
0.967 0.0353 686.4877 1204 2.700 0.051 2.881 0.052 0.937 0.0358
888.7394 1204 5.702 0.080 5.400 0.110 1.056 0.0358 825.6926 1203
1.840 0.110 2.235 0.124 0.823 0.0369
746.557 1202 2.166 0.030 2.057 0.037 1.053 0.0378 757.5625 1201
7.728 0.046 7.909 0.077 0.977 0.0391 615.4798 1204 2.647 0.049
2.827 0.067 0.936 0.0396 831.7408 1203 4.104 0.085 4.425 0.130
0.928 0.0405 761.5846 1201 3.155 0.051 3.357 0.091 0.940 0.0414
581.3344 1202 1.927 0.076 2.207 0.110 0.873 0.0423 1098.9739 1204
2.955 0.090 2.606 0.152 1.134 0.0427 380.3096 1204 1.650 0.070
1.899 0.090 0.869 0.0434 565.3394 1202 7.001 0.052 7.186 0.068
0.974 0.0439 478.3664 1203 1.493 0.068 1.212 0.143 1.232 0.0475
835.7006 1204 2.799 0.115 3.214 0.167 0.871 0.0485 320.2356 1204
1.447 0.070 1.686 0.085 0.858 0.0486 493.385 1203 1.979 0.066 1.722
0.124 1.149 0.049 512.4082 1204 2.415 0.121 1.954 0.215 1.236
0.0493 610.3686 1201 5.156 0.083 4.771 0.223 1.081 0.0495 760.5811
1201 5.225 0.052 5.422 0.091 0.964 0.0495 600.5127 1203 8.858 0.044
8.692 0.074 1.019 0.0496 715.5167 1204 2.592 0.082 2.902 0.141
0.893 0.0498 759.5779 1201 6.347 0.051 6.541 0.090 0.970 0.0499
TABLE-US-00002 TABLE 2 Accurate mass features differing between
clinically diagnosed AD patients with a significant cognitive
impairment and non-demented controls (p < 0.05, log2
transformed). Detected Analysis AVG (log2) SEM AVG (log2) SEM
log(2) Mass Mode AD + Cog AD + Cog Normal Normal Ratio P Value
541.3432 1102 3.138 0.034 3.798 0.048 1.210 3.45E-17 569.3687 1102
2.063 0.042 2.724 0.048 1.320 1.67E-15 723.5197 1204 3.581 0.053
4.350 0.056 1.215 1.04E-14 803.568 1102 5.238 0.048 5.838 0.035
1.115 2.00E-14 804.5713 1102 4.017 0.048 4.610 0.033 1.148 2.02E-14
723.5195 1202 2.193 0.056 2.892 0.052 1.319 4.37E-13 749.5367 1202
3.187 0.044 3.714 0.041 1.165 2.12E-12 555.3102 1102 1.634 0.053
2.240 0.046 1.371 3.10E-12 565.3394 1102 3.265 0.058 3.958 0.055
1.212 3.23E-12 724.5257 1204 2.850 0.048 3.463 0.055 1.215 4.54E-12
699.5198 1204 2.129 0.055 2.775 0.054 1.303 9.18E-12 871.5528 1102
2.885 0.049 3.408 0.040 1.181 2.08E-11 567.3547 1102 2.629 0.050
3.218 0.054 1.224 2.92E-11 751.5555 1204 4.591 0.056 5.248 0.060
1.143 3.18E-11 780.5474 1101 5.241 0.064 5.875 0.043 1.121 3.70E-11
752.5564 1202 2.281 0.049 2.836 0.050 1.244 4.78E-11 779.5444 1101
6.239 0.065 6.874 0.043 1.102 5.01E-11 829.5856 1102 3.806 0.059
4.398 0.046 1.156 9.29E-11 794.5421 1102 2.364 0.048 2.853 0.040
1.206 1.01E-10 793.5386 1102 3.435 0.052 3.950 0.039 1.150 1.05E-10
831.5997 1102 4.372 0.055 4.903 0.040 1.121 1.61E-10 832.6026 1102
3.268 0.055 3.795 0.040 1.161 1.67E-10 751.5529 1202 3.369 0.052
3.920 0.050 1.163 1.89E-10 749.5407 1204 4.423 0.047 5.012 0.064
1.133 1.93E-10 827.57 1102 3.984 0.059 4.630 0.062 1.162 1.95E-10
725.5385 1204 2.815 0.058 3.417 0.054 1.214 2.58E-10 804.5476 1101
3.987 0.073 4.703 0.060 1.180 2.67E-10 837.5027 1101 3.402 0.050
3.933 0.050 1.156 2.76E-10 803.5445 1101 4.941 0.074 5.655 0.059
1.144 3.46E-10 752.5583 1204 3.484 0.056 4.094 0.061 1.175 3.59E-10
828.5737 1102 2.982 0.056 3.590 0.061 1.204 3.85E-10 750.544 1204
3.258 0.054 3.858 0.063 1.184 6.22E-10 750.5402 1202 2.210 0.052
2.704 0.044 1.224 1.19E-09 805.5832 1102 3.874 0.056 4.485 0.068
1.158 1.95E-09 727.5568 1204 3.445 0.055 3.986 0.060 1.157 7.95E-09
807.5758 1101 5.531 0.069 6.102 0.047 1.103 8.66E-09 805.5605 1101
5.184 0.074 5.785 0.048 1.116 9.26E-09 808.5792 1101 4.488 0.071
5.063 0.047 1.128 1.11E-08 806.5639 1101 4.199 0.075 4.779 0.049
1.138 3.65E-08 915.5191 1101 2.189 0.058 2.717 0.063 1.241 5.21E-08
817.5377 1102 2.175 0.064 2.712 0.057 1.247 5.29E-08 781.5617 1101
5.928 0.084 6.610 0.072 1.115 8.47E-08 819.5551 1102 2.198 0.058
2.654 0.046 1.208 9.28E-08 726.5461 1204 2.759 0.044 3.150 0.050
1.141 1.31E-07 783.5672 1101 3.535 0.092 4.259 0.079 1.205 1.69E-07
886.5582 1102 2.911 0.041 3.250 0.041 1.117 2.03E-07 782.565 1101
4.901 0.088 5.589 0.078 1.140 2.35E-07 784.5237 1204 3.211 0.056
3.603 0.037 1.122 4.12E-07 853.5854 1102 2.449 0.056 2.880 0.052
1.176 4.56E-07 795.555 1102 2.520 0.054 2.986 0.065 1.185 5.49E-07
855.6016 1102 3.386 0.060 3.873 0.065 1.144 6.32E-07 821.5712 1102
2.919 0.053 3.304 0.044 1.132 6.68E-07 795.5181 1101 2.458 0.082
3.044 0.064 1.238 7.57E-07 832.5791 1101 3.184 0.076 3.723 0.061
1.169 9.68E-07 786.5416 1204 3.729 0.049 4.087 0.043 1.096 1.02E-06
831.5759 1101 4.123 0.080 4.677 0.060 1.134 1.03E-06 728.5627 1204
2.875 0.047 3.281 0.060 1.141 1.07E-06 757.4991 1101 3.450 0.089
4.073 0.070 1.181 1.08E-06 760.5216 1204 4.008 0.048 4.347 0.039
1.084 1.13E-06 829.5604 1101 3.360 0.075 3.875 0.058 1.153 1.41E-06
847.5316 1101 2.564 0.060 3.091 0.083 1.205 1.84E-06 755.5468 1101
2.115 0.086 2.753 0.085 1.302 1.88E-06 782.5085 1204 3.369 0.056
3.780 0.055 1.122 2.13E-06 755.486 1204 3.146 0.061 3.561 0.057
1.132 5.93E-06 833.5932 1101 3.045 0.095 3.608 0.056 1.185 7.05E-06
758.5656 1101 5.511 0.107 6.160 0.078 1.118 1.11E-05 758.5092 1204
4.504 0.047 4.808 0.042 1.068 1.24E-05 757.5626 1101 6.470 0.106
7.108 0.079 1.099 1.48E-05 760.5811 1101 4.216 0.099 4.886 0.103
1.159 1.48E-05 744.5536 1204 4.260 0.047 4.605 0.058 1.081 1.55E-05
759.5779 1101 5.352 0.096 5.997 0.099 1.121 1.60E-05 811.6096 1101
2.801 0.116 3.511 0.096 1.253 1.84E-05 731.5464 1101 1.634 0.139
2.600 0.157 1.591 1.89E-05 812.5762 1202 1.752 0.045 2.084 0.058
1.190 1.95E-05 743.5471 1204 5.227 0.047 5.579 0.063 1.067 2.40E-05
784.5811 1101 4.161 0.112 4.817 0.089 1.158 3.03E-05 632.5762 1203
0.925 0.149 1.830 0.132 1.979 3.07E-05 761.5843 1101 2.241 0.084
2.829 0.106 1.262 4.28E-05 517.314 1101 5.399 0.050 5.755 0.069
1.066 0.0001 518.3174 1101 3.358 0.051 3.717 0.073 1.107 0.0001
591.3542 1202 3.870 0.092 4.345 0.058 1.123 0.0001 732.4938 1204
4.186 0.056 4.511 0.057 1.078 0.0001 745.5658 1204 3.881 0.052
4.242 0.068 1.093 0.0001 746.5717 1204 3.047 0.043 3.362 0.061
1.103 0.0001 747.5245 1204 3.433 0.058 3.886 0.090 1.132 0.0001
777.553 1202 1.720 0.100 2.384 0.130 1.386 0.0001 783.5148 1204
3.217 0.063 3.574 0.056 1.111 0.0001 785.5933 1101 5.687 0.112
6.271 0.080 1.103 0.0001 786.5967 1101 4.611 0.112 5.203 0.078
1.128 0.0001 809.5936 1101 4.786 0.104 5.362 0.079 1.120 0.0001
810.5969 1101 3.779 0.108 4.370 0.080 1.157 0.0001 811.5732 1202
2.729 0.047 3.027 0.055 1.109 0.0001 858.6212 1202 2.533 0.080
2.994 0.074 1.182 0.0001 306.2569 1204 2.878 0.070 3.266 0.067
1.135 0.0002 482.3215 1202 1.940 0.050 2.251 0.062 1.160 0.0002
522.4635 1203 4.489 0.181 5.485 0.176 1.222 0.0002 523.4679 1203
3.035 0.180 4.011 0.165 1.322 0.0002 579.5325 1203 3.962 0.112
4.636 0.127 1.170 0.0002 759.5163 1204 4.645 0.061 4.954 0.048
1.066 0.0002 787.5729 1202 1.834 0.054 2.145 0.057 1.170 0.0002
810.5399 1204 2.760 0.071 3.145 0.066 1.140 0.0002 814.5917 1202
2.054 0.035 2.289 0.051 1.114 0.0002 857.6186 1202 3.614 0.081
4.049 0.076 1.121 0.0002 458.2405 1101 1.632 0.054 1.986 0.077
1.217 0.0003 481.3172 1202 3.866 0.050 4.159 0.060 1.076 0.0003
541.3139 1101 4.022 0.081 4.494 0.096 1.117 0.0003 545.3453 1101
3.500 0.071 3.898 0.078 1.114 0.0003 592.3571 1202 2.113 0.095
2.560 0.062 1.212 0.0003 757.5014 1204 3.642 0.067 3.971 0.052
1.091 0.0003 833.7551 1204 2.061 0.223 3.140 0.161 1.524 0.0003
543.3296 1101 3.652 0.055 4.023 0.087 1.102 0.0004 605.5457 1203
5.025 0.091 5.509 0.089 1.096 0.0004 731.4916 1204 5.419 0.060
5.746 0.063 1.060 0.0004 775.5533 1202 2.106 0.063 2.449 0.068
1.163 0.0004 813.5885 1202 3.034 0.043 3.276 0.048 1.080 0.0004
542.3173 1101 2.234 0.087 2.687 0.089 1.203 0.0005 722.5335 1101
2.406 0.062 2.717 0.056 1.129 0.0005 744.55 1202 1.842 0.064 2.222
0.083 1.206 0.0005 769.5656 1204 3.889 0.055 4.174 0.054 1.073
0.0005 807.59 1202 2.348 0.061 2.718 0.082 1.158 0.0005 828.5743
1202 5.181 0.076 5.598 0.085 1.081 0.0005 521.4522 1203 2.124 0.191
3.028 0.158 1.425 0.0007 604.5433 1203 6.440 0.091 6.901 0.089
1.072 0.0007 744.4956 1204 3.851 0.049 4.117 0.058 1.069 0.0007
748.5287 1204 2.249 0.127 2.820 0.088 1.254 0.0007 771.5814 1204
4.118 0.051 4.367 0.046 1.061 0.0007 827.5701 1202 6.245 0.082
6.684 0.092 1.070 0.0007 747.5201 1202 1.886 0.083 2.313 0.088
1.226 0.0008 775.5528 1204 3.176 0.061 3.537 0.085 1.114 0.0008
787.5465 1204 2.543 0.116 3.017 0.054 1.187 0.0008 827.5448 1101
3.373 0.104 3.873 0.095 1.148 0.0008 546.3485 1101 1.890 0.073
2.287 0.089 1.210 0.0009 520.4499 1203 3.775 0.176 4.577 0.148
1.212 0.001 570.3725 1202 2.926 0.047 3.155 0.045 1.078 0.001
781.562 1201 7.334 0.054 7.638 0.071 1.041 0.001 632.5032 1203
1.435 0.112 0.795 0.154 0.554 0.0011 743.5469 1202 2.976 0.070
3.348 0.086 1.125 0.0012 768.5539 1204 3.928 0.069 4.279 0.077
1.089 0.0012 806.5873 1202 4.280 0.057 4.635 0.092 1.083 0.0012
575.2728 1101 2.419 0.061 2.739 0.074 1.132 0.0013 550.4957 1203
6.766 0.117 7.415 0.160 1.096 0.0014 805.5839 1202 5.470 0.060
5.841 0.097 1.068 0.0014 183.0661 1101 2.412 0.147 3.015 0.094
1.250 0.0015 551.4986 1203 5.350 0.117 5.988 0.158 1.119 0.0015
741.5305 1204 2.825 0.080 3.250 0.102 1.150 0.0015 541.3435 1202
5.595 0.072 5.974 0.094 1.068 0.0018 552.5022 1203 3.055 0.110
3.643 0.147 1.193 0.0018 506.3213 1202 2.487 0.054 2.748 0.061
1.105 0.0019 569.369 1202 4.867 0.049 5.089 0.047 1.046 0.0019
782.5653 1201 6.310 0.058 6.614 0.076 1.048 0.002 490.3641 1203
2.028 0.121 1.397 0.159 0.689 0.0021 542.3461 1202 3.803 0.061
4.106 0.074 1.080 0.0021 566.3434 1202 5.130 0.071 5.436 0.062
1.060 0.0022 833.7571 1203 2.853 0.122 3.507 0.170 1.229 0.0022
837.718 1204 2.593 0.250 3.662 0.217 1.412 0.0023 549.4845 1203
5.510 0.107 5.994 0.110 1.088 0.0025 793.5681 1204 3.116 0.058
3.372 0.055 1.082 0.0025 478.254 1201 1.629 0.116 1.051 0.148 0.645
0.0027 536.4794 1203 2.188 0.134 2.799 0.143 1.279 0.0027 548.4815
1203 6.949 0.103 7.405 0.103 1.066 0.0027 832.6037 1202 4.981 0.069
5.274 0.062 1.059 0.0028 830.5894 1202 4.738 0.076 5.051 0.064
1.066 0.0029 1098.9739 1204 3.097 0.070 2.606 0.152 0.842 0.0031
767.5495 1204 5.033 0.074 5.369 0.082 1.067 0.0034 675.6377 1204
3.875 0.066 4.160 0.066 1.074 0.0035 564.5134 1203 2.641 0.093
3.075 0.111 1.164 0.0036 789.5892 1202 1.703 0.053 1.952 0.065
1.147 0.0036 832.7523 1203 3.734 0.131 4.374 0.169 1.171 0.0036
748.5735 1202 3.940 0.052 3.722 0.050 0.945 0.0037 801.555 1202
2.543 0.077 2.881 0.082 1.133 0.0037 856.672 1202 2.777 0.048 2.553
0.057 0.919 0.0037 858.6861 1202 2.982 0.055 2.756 0.052 0.924
0.0043 803.5677 1202 7.101 0.095 7.502 0.098 1.057 0.0045 838.7226
1204 2.127 0.200 2.973 0.206 1.398 0.0046 685.26 1202 1.708 0.074
1.998 0.063 1.169 0.0047 826.7069 1204 2.279 0.148 2.819 0.099
1.237 0.0047 582.2473 1201 3.332 0.122 2.793 0.141 0.838 0.0049
590.343 1202 4.005 0.069 4.304 0.076 1.075 0.005 832.7492 1204
3.163 0.146 3.848 0.190 1.217 0.0051 829.5859 1202 5.864 0.082
6.181 0.070 1.054 0.0053 772.5862 1204 3.263 0.043 3.437 0.042
1.053 0.0054 409.0208 1202 2.753 0.050 2.980 0.063 1.082 0.0055
509.3493 1202 2.358 0.054 2.579 0.055 1.094 0.0056 589.3403 1202
5.847 0.076 6.171 0.085 1.056 0.0057 430.3818 1204 4.790 0.109
5.253 0.122 1.097 0.0061 804.5718 1202 5.801 0.078 6.117 0.080
1.054 0.0066 505.3229 1202 3.931 0.071 4.222 0.076 1.074 0.0068
523.3634 1101 3.365 0.111 3.791 0.103 1.127 0.0072 671.5723 1204
2.131 0.140 2.604 0.087 1.222 0.0073 830.7352 1204 4.091 0.130
4.638 0.150 1.134 0.0073 431.386 1204 3.354 0.108 3.795 0.118 1.132
0.0074 631.628 1203 1.746 0.166 2.391 0.161 1.369 0.0075 825.6926
1203 1.644 0.169 2.235 0.124 1.359 0.0079 614.4914 1203 2.821 0.080
2.448 0.114 0.868 0.008 831.6001 1202 6.109 0.077 6.404 0.074 1.048
0.0081 615.3539 1202 2.394 0.063 2.629 0.058 1.098 0.0083 568.3573
1202 3.959 0.049 4.167 0.060 1.053 0.0088 767.547 1202 2.992 0.097
3.358 0.092 1.122 0.0088 768.5503 1202 1.849 0.125 2.277 0.090
1.231 0.0089 831.7408 1203 3.971 0.109 4.425 0.130 1.114 0.0089
411.3212 1202 2.838 0.054 3.033 0.048 1.069 0.0099 796.5876 1204
2.633 0.059 2.860 0.062 1.087 0.0099 863.6876 1204 4.853 0.087
5.188 0.092 1.069 0.0104 492.3816 1203 3.588 0.076 3.302 0.077
0.920 0.0105 772.5278 1204 3.282 0.056 3.488 0.055 1.063 0.0107
825.5544 1202 2.644 0.112 3.084 0.126 1.167 0.0108 320.2356 1204
1.302 0.114 1.686 0.085 1.295 0.011 380.3096 1204 1.580 0.083 1.899
0.090 1.202 0.0112 519.3321 1101 3.914 0.122 4.382 0.133 1.119
0.0116 711.2577 1202 2.205 0.075 2.454 0.056 1.113 0.0118 493.385
1203 2.083 0.073 1.722 0.124 0.827 0.0119 565.3394 1202 6.924 0.073
7.186 0.068 1.038 0.0119 670.569 1204 3.179 0.090 3.515 0.093 1.106
0.012 856.6061 1202 4.500 0.081 4.805 0.086 1.068 0.0121 340.2976
1203 1.502 0.124 1.937 0.112 1.290 0.0124 287.2824 1101 1.958 0.180
1.265 0.202 0.646 0.0126 495.332 1101 5.196 0.094 5.565 0.112 1.071
0.0137 304.241 1204 4.876 0.053 5.088 0.066 1.044 0.0138 305.2438
1204 2.492 0.061 2.719 0.067 1.091 0.0145 616.5052 1203 4.260 0.086
3.957 0.084 0.929 0.0145 746.5119 1204 2.615 0.165 3.137 0.119
1.200 0.0152 861.7806 1203 2.533 0.092 2.902 0.119 1.146 0.0152
686.4877 1204 2.617 0.088 2.881 0.052 1.101 0.0155 830.7363 1203
4.834 0.111 5.263 0.135 1.089 0.0162 835.7006 1204 2.651 0.155
3.214 0.167 1.212 0.0165 867.7579 1204 2.654 0.185 3.308 0.189
1.246 0.0167 870.7307 1203 3.361 0.074 2.985 0.142 0.888 0.0169
246.1465 1202 3.761 0.084 4.072 0.096 1.083 0.017 507.3316 1202
2.891 0.047 3.094 0.071 1.070 0.0173 855.6023 1202 5.497 0.079
5.784 0.087 1.052 0.0173 578.5277 1203 4.183 0.420 5.548 0.355
1.326 0.0176 615.4938 1203 1.386 0.133 0.902 0.148 0.651 0.0177
808.5803 1201 5.667 0.029 5.784 0.039 1.021 0.0177 860.7752 1204
3.643 0.124 4.070 0.125 1.117 0.0185 518.4345 1203 1.584 0.180
2.199 0.178 1.388 0.0186 520.3354 1101 2.215 0.123 2.646 0.130
1.195 0.0188 765.5313 1202 1.717 0.123 2.098 0.095 1.222 0.0195
777.5287 1201 2.793 0.063 3.020 0.072 1.082 0.0198 887.8005 1203
2.948 0.118 3.306 0.086 1.122 0.0198 739.5143 1204 2.602 0.149
3.121 0.158 1.200 0.0199 831.7387 1204 3.193 0.121 3.648 0.150
1.142 0.02
521.3477 1101 3.643 0.099 4.021 0.127 1.104 0.0203 584.2646 1204
3.154 0.130 2.649 0.172 0.840 0.0207 661.6233 1204 2.430 0.088
2.715 0.079 1.117 0.0209 715.5167 1204 2.449 0.131 2.902 0.141
1.185 0.0218 529.3167 1202 3.031 0.071 3.265 0.069 1.077 0.0221
544.4481 1203 2.105 0.067 2.333 0.071 1.108 0.0234 866.7532 1204
4.149 0.145 4.693 0.189 1.131 0.0235 581.3344 1202 1.844 0.111
2.207 0.110 1.197 0.0245 795.5838 1204 3.569 0.060 3.769 0.063
1.056 0.0246 807.5768 1201 6.720 0.030 6.829 0.037 1.016 0.0256
854.5902 1202 3.801 0.070 4.031 0.073 1.060 0.0266 865.7487 1204
4.895 0.152 5.445 0.193 1.112 0.0268 496.3355 1101 3.396 0.095
3.736 0.119 1.100 0.027 755.5467 1201 3.507 0.072 3.755 0.084 1.071
0.0274 1019.3838 1102 3.035 0.036 2.919 0.037 0.962 0.0291 684.5489
1204 2.508 0.071 2.745 0.080 1.094 0.0294 774.0316 1204 1.789 0.192
1.167 0.203 0.652 0.0294 765.5334 1204 3.218 0.101 3.540 0.104
1.100 0.0299 630.5587 1203 3.126 0.072 3.344 0.064 1.070 0.0301
488.3873 1203 2.241 0.069 1.986 0.096 0.886 0.0321 302.2255 1204
3.506 0.085 3.765 0.081 1.074 0.0322 757.5625 1201 7.689 0.066
7.909 0.077 1.029 0.0325 617.5089 1203 2.978 0.089 2.698 0.092
0.906 0.0328 829.7239 1204 2.802 0.120 3.185 0.131 1.137 0.0344
303.1079 1202 5.656 0.048 5.802 0.047 1.026 0.0348 826.5581 1202
1.648 0.107 2.032 0.149 1.233 0.0362 506.2851 1201 3.080 0.117
2.656 0.165 0.862 0.0363 504.3814 1203 1.746 0.098 1.413 0.124
0.809 0.0366 691.1957 1102 2.130 0.060 1.938 0.068 0.910 0.0373
626.5278 1203 3.820 0.064 4.009 0.060 1.049 0.0375 522.3511 1101
1.753 0.091 2.069 0.121 1.180 0.0385 759.5779 1201 6.294 0.076
6.541 0.090 1.039 0.0386 625.5161 1203 2.856 0.057 3.045 0.070
1.066 0.0387 760.5811 1201 5.173 0.077 5.422 0.091 1.048 0.0395
484.3794 1204 2.181 0.092 1.776 0.178 0.815 0.0396 819.5642 1202
2.811 0.080 3.104 0.120 1.104 0.0404 853.5862 1202 4.775 0.069
4.985 0.075 1.044 0.0423 700.552 1101 2.278 0.124 2.617 0.102 1.149
0.0426 709.2594 1202 2.152 0.092 2.382 0.053 1.107 0.0426 662.5175
1204 3.806 0.120 3.420 0.145 0.899 0.0432 761.5846 1201 3.118 0.074
3.357 0.091 1.077 0.0436 743.5466 1203 2.011 0.077 2.266 0.099
1.127 0.0442 478.3664 1203 1.545 0.088 1.212 0.143 0.785 0.0447
784.5811 1201 5.585 0.073 5.809 0.082 1.040 0.0448 860.7756 1203
4.383 0.087 4.653 0.100 1.061 0.0448 601.5164 1203 7.642 0.056
7.463 0.069 0.976 0.0453 758.5655 1201 6.637 0.074 6.860 0.084
1.034 0.0488 371.3542 1203 3.253 0.055 3.435 0.074 1.056 0.0489
783.5778 1201 6.661 0.074 6.883 0.083 1.033 0.049 921.8145 1204
2.716 0.211 3.255 0.155 1.198 0.0494 824.6892 1203 2.657 0.108
2.964 0.108 1.116 0.0497
TABLE-US-00003 TABLE 3 Accurate mass features differing between
clinically diagnosed AD patients with a significant cognitive
impairment and clinically diagnosed non-AD patients with a
significant cognitive impairment (p < 0.05, log2 transformed).
Detected Analysis AVG (log2) SEM AVG (log2) SEM log(2) Mass Mode AD
+ Cog AD + Cog Non-AD + Cog Non-AD + Cog Ratio P Value 723.5197
1204 3.581 0.053 4.571 0.070 0.783 1.07E-17 723.5195 1202 2.193
0.056 3.033 0.049 0.723 2.70E-17 749.5367 1202 3.187 0.044 3.812
0.045 0.836 5.25E-15 724.5257 1204 2.850 0.048 3.630 0.066 0.785
1.68E-14 752.5564 1202 2.281 0.049 2.975 0.056 0.767 5.71E-14
751.5555 1204 4.591 0.056 5.427 0.071 0.846 6.78E-14 751.5529 1202
3.369 0.052 4.055 0.054 0.831 1.46E-13 752.5583 1204 3.484 0.056
4.284 0.070 0.813 2.38E-13 749.5407 1204 4.423 0.047 5.185 0.073
0.853 5.02E-13 750.5402 1202 2.210 0.052 2.804 0.044 0.788 9.50E-13
750.544 1204 3.258 0.054 4.004 0.074 0.814 7.49E-12 725.5385 1204
2.815 0.058 3.543 0.078 0.794 1.10E-10 699.5198 1204 2.129 0.055
2.825 0.079 0.754 3.41E-10 727.5568 1204 3.445 0.055 4.039 0.076
0.853 1.48E-08 728.5627 1204 2.875 0.047 3.301 0.056 0.871 1.21E-07
726.5461 1204 2.759 0.044 3.175 0.057 0.869 1.35E-07 775.5528 1204
3.176 0.061 3.712 0.075 0.856 3.75E-07 813.5885 1202 3.034 0.043
3.369 0.049 0.901 2.08E-06 775.5533 1202 2.106 0.063 2.524 0.056
0.834 4.26E-06 747.5245 1204 3.433 0.058 3.933 0.088 0.873 8.18E-06
814.5917 1202 2.054 0.035 2.355 0.055 0.872 1.16E-05 747.5201 1202
1.886 0.083 2.357 0.062 0.800 2.25E-05 811.5732 1202 2.729 0.047
3.041 0.055 0.897 4.54E-05 789.5892 1202 1.703 0.053 2.013 0.061
0.846 0.0002 810.5399 1204 2.760 0.071 3.111 0.064 0.887 0.0004
795.5838 1204 3.569 0.060 3.845 0.050 0.928 0.0007 856.6061 1202
4.500 0.081 4.894 0.078 0.919 0.0008 783.5148 1204 3.217 0.063
3.512 0.061 0.916 0.0011 855.6023 1202 5.497 0.079 5.876 0.079
0.935 0.0011 858.6212 1202 2.533 0.080 2.917 0.080 0.868 0.0011
787.5465 1204 2.543 0.116 2.964 0.045 0.858 0.0013 857.6186 1202
3.614 0.081 3.989 0.081 0.906 0.0015 773.537 1202 1.581 0.066 1.875
0.065 0.843 0.002 784.5237 1204 3.211 0.056 3.464 0.056 0.927 0.002
748.5287 1204 2.249 0.127 2.811 0.123 0.800 0.0021 828.5743 1202
5.181 0.076 5.517 0.075 0.939 0.0023 827.5701 1202 6.245 0.082
6.608 0.083 0.945 0.0026 786.5416 1204 3.729 0.049 3.961 0.058
0.941 0.0029 871.5934 1202 1.841 0.085 2.189 0.076 0.841 0.0031
744.5536 1204 4.260 0.047 4.473 0.054 0.952 0.0037 787.5729 1202
1.834 0.054 2.081 0.065 0.881 0.0042 755.486 1204 3.146 0.061 3.395
0.059 0.927 0.0044 796.5876 1204 2.633 0.059 2.868 0.056 0.918
0.0048 812.5762 1202 1.752 0.045 2.034 0.088 0.861 0.0049 817.5377
1102 2.175 0.064 2.427 0.060 0.896 0.0053 383.3284 1204 1.263 0.120
1.722 0.107 0.733 0.0055 840.6063 1202 2.746 0.042 2.927 0.050
0.938 0.0065 544.397 1204 2.998 0.111 2.235 0.255 1.342 0.0066
570.3725 1202 2.926 0.047 3.115 0.049 0.939 0.0071 782.5085 1204
3.369 0.056 3.596 0.062 0.937 0.0077 847.5954 1202 2.317 0.089
2.662 0.090 0.870 0.0079 855.6016 1102 3.386 0.060 3.619 0.062
0.936 0.0082 769.5656 1204 3.889 0.055 4.103 0.057 0.948 0.0084
819.5642 1202 2.811 0.080 3.149 0.099 0.893 0.0089 828.5737 1102
2.982 0.056 3.197 0.058 0.933 0.009 590.343 1202 4.005 0.069 4.261
0.066 0.940 0.0091 719.6231 1204 4.528 0.074 4.158 0.119 1.089
0.0092 589.3403 1202 5.847 0.076 6.129 0.075 0.954 0.0098 768.5539
1204 3.928 0.069 4.189 0.071 0.938 0.0102 839.6031 1202 3.723 0.041
3.895 0.051 0.956 0.0102 821.5712 1102 2.919 0.053 3.120 0.056
0.936 0.0106 1226.097 1203 3.278 0.092 2.921 0.105 1.122 0.0122
664.5323 1204 4.909 0.126 4.391 0.161 1.118 0.0126 733.6426 1204
2.981 0.074 2.625 0.120 1.135 0.0126 771.5814 1204 4.118 0.051
4.313 0.057 0.955 0.0126 809.5937 1201 6.453 0.060 6.677 0.065
0.966 0.0126 720.6258 1204 3.565 0.076 3.215 0.117 1.109 0.0127
667.5475 1204 3.516 0.124 3.013 0.157 1.167 0.0132 665.5354 1204
3.705 0.121 3.182 0.170 1.164 0.0133 666.5456 1204 4.713 0.126
4.232 0.146 1.114 0.0142 793.5386 1102 3.435 0.052 3.632 0.061
0.946 0.015 810.5971 1201 5.436 0.063 5.665 0.068 0.959 0.015
832.6037 1202 4.981 0.069 5.231 0.074 0.952 0.0151 663.5216 1204
2.566 0.145 1.946 0.210 1.319 0.0159 743.5471 1204 5.227 0.047
5.417 0.062 0.965 0.0162 767.5495 1204 5.033 0.074 5.287 0.074
0.952 0.017 796.5292 1204 2.928 0.063 3.142 0.062 0.932 0.017
671.5723 1204 2.131 0.140 2.568 0.113 0.830 0.0173 758.5092 1204
4.504 0.047 4.670 0.051 0.964 0.0188 509.3493 1202 2.358 0.054
2.542 0.056 0.927 0.0192 827.57 1102 3.984 0.059 4.191 0.064 0.951
0.0194 569.369 1202 4.867 0.049 5.033 0.049 0.967 0.02 831.6001
1202 6.109 0.077 6.380 0.085 0.958 0.02 638.5149 1204 3.530 0.120
3.086 0.150 1.144 0.0224 768.5503 1202 1.849 0.125 2.190 0.080
0.845 0.0249 313.1153 1204 3.859 0.102 3.338 0.209 1.156 0.025
592.3571 1202 2.113 0.095 2.375 0.065 0.890 0.0262 794.5421 1102
2.364 0.048 2.536 0.061 0.932 0.0276 662.5175 1204 3.806 0.120
3.377 0.153 1.127 0.0292 670.569 1204 3.179 0.090 3.474 0.101 0.915
0.0312 870.7307 1203 3.361 0.074 3.012 0.144 1.116 0.0312 1098.974
1204 3.097 0.070 2.739 0.151 1.131 0.0313 694.6161 1204 2.845 0.080
2.437 0.173 1.168 0.0324 541.3432 1102 3.138 0.034 3.255 0.043
0.964 0.0335 581.3344 1202 1.844 0.111 2.147 0.087 0.859 0.0351
691.596 1204 2.394 0.098 1.999 0.159 1.198 0.0351 803.5677 1202
7.101 0.095 7.405 0.108 0.959 0.0364 591.3542 1202 3.870 0.092
4.110 0.064 0.942 0.0367 705.6093 1204 3.137 0.072 2.823 0.132
1.111 0.0367 847.5316 1101 2.564 0.060 2.815 0.104 0.911 0.0367
854.5902 1202 3.801 0.070 4.027 0.082 0.944 0.0376 215.9153 1101
4.405 0.297 5.221 0.247 0.844 0.0379 569.3687 1102 2.063 0.042
2.217 0.062 0.931 0.0404 640.5294 1204 3.725 0.127 3.313 0.154
1.124 0.0407 853.5862 1202 4.775 0.069 4.993 0.081 0.956 0.0413
830.5894 1202 4.738 0.076 4.968 0.082 0.954 0.0415 760.5216 1204
4.008 0.048 4.156 0.053 0.964 0.0417 530.3821 1204 2.614 0.172
2.016 0.240 1.296 0.0441 1225.092 1203 3.451 0.135 3.011 0.170
1.146 0.0445 819.5551 1102 2.198 0.058 2.369 0.062 0.928 0.0456
743.5469 1202 2.976 0.070 3.200 0.087 0.930 0.0464 444.2717 1202
1.304 0.083 1.577 0.109 0.827 0.0471 678.5477 1204 3.874 0.101
3.523 0.145 1.099 0.0485 741.5305 1204 2.825 0.080 3.086 0.104
0.916 0.0489 759.5163 1204 4.645 0.061 4.811 0.057 0.966 0.0493
820.568 1202 1.757 0.128 2.102 0.117 0.836 0.0497
TABLE-US-00004 TABLE 4 Accurate mass features differing between
clinically diagnosed AD patients with a significant cognitive
impairment and clinically diagnosed AD patients without a
significant cognitive impairment (p < 0.05, log2 transformed).
Detected Analysis AVG (log2) SEM AVG (log2) SEM log(2) Mass Mode AD
AD AD noCog AD noCog Ratio P Value 541.3432 1102 3.138 0.034 3.518
0.039 0.892 3.47E-10 569.3687 1102 2.063 0.042 2.479 0.045 0.832
3.90E-09 829.5856 1102 3.806 0.059 4.301 0.048 0.885 1.30E-08
567.3547 1102 2.629 0.050 3.031 0.045 0.868 1.02E-07 831.5997 1102
4.372 0.055 4.774 0.042 0.916 1.93E-07 832.6026 1102 3.268 0.055
3.662 0.042 0.892 2.67E-07 804.5713 1102 4.017 0.048 4.426 0.054
0.908 3.40E-07 803.568 1102 5.238 0.048 5.656 0.057 0.926 3.97E-07
871.5528 1102 2.885 0.049 3.279 0.054 0.880 8.30E-07 565.3394 1102
3.265 0.058 3.728 0.064 0.876 1.14E-06 805.5832 1102 3.874 0.056
4.292 0.055 0.903 1.41E-06 819.5551 1102 2.198 0.058 2.604 0.053
0.844 2.52E-06 555.3102 1102 1.634 0.053 2.033 0.059 0.803 3.32E-06
853.5854 1102 2.449 0.056 2.824 0.051 0.867 5.09E-06 805.5605 1101
5.184 0.074 5.665 0.065 0.915 6.42E-06 808.5792 1101 4.488 0.071
4.925 0.057 0.911 9.48E-06 793.5386 1102 3.435 0.052 3.799 0.057
0.904 1.17E-05 807.5758 1101 5.531 0.069 5.961 0.059 0.928 1.20E-05
806.5639 1101 4.199 0.075 4.669 0.066 0.899 1.28E-05 837.5027 1101
3.402 0.050 3.781 0.064 0.900 1.45E-05 794.5421 1102 2.364 0.048
2.703 0.059 0.875 2.79E-05 780.5474 1101 5.241 0.064 5.661 0.070
0.926 3.62E-05 779.5444 1101 6.239 0.065 6.654 0.070 0.938 4.37E-05
821.5712 1102 2.919 0.053 3.221 0.048 0.906 0.0001 827.57 1102
3.984 0.059 4.337 0.063 0.919 0.0001 886.5582 1102 2.911 0.041
3.189 0.053 0.913 0.0001 915.5191 1101 2.189 0.058 2.592 0.073
0.845 0.0001 722.5335 1101 2.406 0.062 2.735 0.057 0.880 0.0002
828.5737 1102 2.982 0.056 3.313 0.063 0.900 0.0002 833.5932 1101
3.045 0.095 3.522 0.077 0.865 0.0002 855.6016 1102 3.386 0.060
3.704 0.055 0.914 0.0002 755.5468 1101 2.115 0.086 2.563 0.082
0.825 0.0003 795.555 1102 2.520 0.054 2.818 0.057 0.894 0.0003
803.5445 1101 4.941 0.074 5.324 0.070 0.928 0.0003 811.6096 1101
2.801 0.116 3.370 0.096 0.831 0.0003 804.5476 1101 3.987 0.073
4.370 0.073 0.912 0.0004 829.5604 1101 3.360 0.075 3.728 0.063
0.901 0.0004 783.5672 1101 3.535 0.092 3.993 0.085 0.885 0.0005
832.5791 1101 3.184 0.076 3.541 0.067 0.899 0.0008 847.5316 1101
2.564 0.060 2.955 0.095 0.868 0.0008 784.5811 1101 4.161 0.112
4.668 0.093 0.891 0.0009 831.5759 1101 4.123 0.080 4.485 0.066
0.919 0.0009 757.4991 1101 3.450 0.089 3.866 0.083 0.892 0.0011
781.5617 1101 5.928 0.084 6.305 0.075 0.940 0.0014 795.5181 1101
2.458 0.082 2.823 0.073 0.871 0.0015 782.565 1101 4.901 0.088 5.279
0.080 0.928 0.0022 760.5811 1101 4.216 0.099 4.632 0.089 0.910
0.0027 759.5779 1101 5.352 0.096 5.752 0.085 0.930 0.0028 591.3542
1202 3.870 0.092 4.224 0.068 0.916 0.0031 592.3571 1202 2.113 0.095
2.472 0.070 0.855 0.0035 761.5843 1101 2.241 0.084 2.609 0.088
0.859 0.0035 858.6212 1202 2.533 0.080 2.870 0.078 0.883 0.0037
458.2405 1101 1.632 0.054 1.858 0.054 0.878 0.0042 785.5933 1101
5.687 0.112 6.096 0.083 0.933 0.0047 786.5967 1101 4.611 0.112
5.018 0.084 0.919 0.005 758.5656 1101 5.511 0.107 5.915 0.094 0.932
0.0061 306.2569 1204 2.878 0.070 3.125 0.053 0.921 0.0066 857.6186
1202 3.614 0.081 3.925 0.076 0.921 0.0066 757.5626 1101 6.470 0.106
6.861 0.094 0.943 0.0075 809.5936 1101 4.786 0.104 5.150 0.081
0.929 0.0077 810.5969 1101 3.779 0.108 4.144 0.083 0.912 0.0095
807.59 1202 2.348 0.061 2.574 0.062 0.912 0.0113 383.3284 1204
1.263 0.120 1.659 0.095 0.761 0.012 786.5416 1204 3.729 0.049 3.903
0.046 0.955 0.012 744.55 1202 1.842 0.064 2.093 0.074 0.880 0.0124
827.5448 1101 3.373 0.104 3.751 0.105 0.899 0.0128 784.5237 1204
3.211 0.056 3.405 0.052 0.943 0.0133 746.5119 1204 2.615 0.165
3.058 0.057 0.855 0.0145 817.5377 1102 2.175 0.064 2.405 0.066
0.905 0.0155 699.5198 1204 2.129 0.055 2.309 0.047 0.922 0.0161
757.5014 1204 3.642 0.067 3.869 0.063 0.941 0.0161 826.7069 1204
2.279 0.148 2.684 0.086 0.849 0.0215 758.5092 1204 4.504 0.047
4.652 0.043 0.968 0.0235 546.3485 1101 1.890 0.073 2.115 0.064
0.894 0.0236 801.555 1202 2.543 0.077 2.788 0.077 0.912 0.0272
829.7239 1204 2.802 0.120 3.161 0.105 0.886 0.0272 748.5721 1102
3.795 0.067 3.996 0.060 0.950 0.0288 518.3174 1101 3.358 0.051
3.523 0.054 0.953 0.0296 826.5581 1202 1.648 0.107 1.983 0.108
0.831 0.0309 830.5894 1202 4.738 0.076 4.956 0.064 0.956 0.0317
430.3818 1204 4.790 0.109 5.108 0.096 0.938 0.0322 313.1153 1204
3.859 0.102 3.311 0.233 1.166 0.0327 755.5467 1201 3.507 0.072
3.728 0.072 0.941 0.0327 545.3453 1101 3.500 0.071 3.713 0.069
0.943 0.0353 320.2356 1204 1.302 0.114 1.590 0.070 0.819 0.0363
183.0661 1101 2.412 0.147 2.779 0.096 0.868 0.0416 825.5544 1202
2.644 0.112 2.968 0.109 0.891 0.0418 549.4845 1203 5.510 0.107
5.818 0.103 0.947 0.0424 829.5859 1202 5.864 0.082 6.087 0.070
0.963 0.0433 431.386 1204 3.354 0.108 3.651 0.099 0.919 0.0463
784.5811 1201 5.585 0.073 5.782 0.064 0.966 0.0463 777.5287 1201
2.793 0.063 2.986 0.072 0.935 0.0466 278.2254 1204 4.828 0.056
4.996 0.062 0.966 0.0467 548.4815 1203 6.949 0.103 7.233 0.096
0.961 0.0472 777.553 1202 1.720 0.100 2.002 0.098 0.859 0.0483
517.314 1101 5.399 0.050 5.547 0.055 0.973 0.0488 661.6233 1204
2.430 0.088 2.638 0.054 0.921 0.0489
TABLE-US-00005 TABLE 5 Accurate mass features differing between
clinically diagnosed non-AD patients and non-demented controls (p
< 0.05, log2 transformed). Detected Analysis AVG (log2) SEM AVG
(log2) SEM log(2) Mass Mode NON-AD NON-AD Normal Normal Ratio P
Value 541.3432 1102 3.255 0.043 3.679 0.048 0.885 3.43E-12 567.3547
1102 2.621 0.052 3.117 0.054 0.841 2.78E-11 805.5605 1101 5.159
0.072 5.604 0.048 0.921 1.08E-09 806.5639 1101 4.150 0.073 4.630
0.049 0.896 1.51E-09 804.5713 1102 4.159 0.056 4.466 0.033 0.931
3.79E-09 783.5672 1101 3.515 0.077 4.126 0.079 0.852 4.13E-09
781.5617 1101 5.919 0.072 6.403 0.072 0.924 4.19E-09 780.5474 1101
5.276 0.075 5.691 0.043 0.927 4.35E-09 803.568 1102 5.379 0.056
5.656 0.035 0.951 4.43E-09 779.5444 1101 6.291 0.073 6.659 0.043
0.945 5.24E-09 760.5811 1101 4.034 0.079 4.733 0.103 0.852 6.28E-09
805.5832 1102 3.897 0.057 4.344 0.068 0.897 6.49E-09 759.5779 1101
5.166 0.082 5.810 0.099 0.889 1.04E-08 782.565 1101 4.878 0.077
5.415 0.078 0.901 1.14E-08 829.5856 1102 3.952 0.051 4.261 0.046
0.927 1.44E-08 569.3687 1102 2.217 0.062 2.639 0.048 0.840 1.99E-08
755.5468 1101 1.974 0.089 2.667 0.085 0.740 2.63E-08 757.4991 1101
3.368 0.086 3.945 0.070 0.854 2.69E-08 871.5528 1102 2.960 0.059
3.301 0.040 0.897 4.58E-08 804.5476 1101 4.125 0.073 4.556 0.060
0.906 6.76E-08 803.5445 1101 5.084 0.072 5.478 0.059 0.928 7.46E-08
758.5656 1101 5.440 0.089 5.968 0.078 0.912 8.14E-08 565.3394 1102
3.359 0.081 3.834 0.055 0.876 8.59E-08 757.5626 1101 6.417 0.087
6.886 0.079 0.932 1.74E-07 808.5792 1101 4.527 0.078 4.904 0.047
0.923 2.30E-07 795.5181 1101 2.491 0.071 2.948 0.064 0.845 2.65E-07
807.5758 1101 5.569 0.080 5.911 0.047 0.942 3.99E-07 731.5464 1101
1.460 0.132 2.519 0.157 0.580 4.14E-07 837.5027 1101 3.437 0.071
3.810 0.050 0.902 4.34E-07 761.5843 1101 2.053 0.092 2.740 0.106
0.749 5.01E-07 784.5811 1101 4.092 0.095 4.666 0.089 0.877 5.41E-07
831.5997 1102 4.512 0.057 4.749 0.040 0.950 6.07E-07 832.6026 1102
3.409 0.056 3.677 0.040 0.927 6.72E-07 555.3102 1102 1.792 0.066
2.170 0.046 0.826 8.11E-07 833.5932 1101 3.038 0.087 3.495 0.056
0.869 1.09E-06 829.5604 1101 3.381 0.077 3.754 0.058 0.901 3.91E-06
811.6096 1101 2.826 0.097 3.401 0.096 0.831 4.19E-06 785.5933 1101
5.629 0.100 6.075 0.080 0.927 5.08E-06 786.5967 1101 4.566 0.101
5.041 0.078 0.906 5.43E-06 827.57 1102 4.191 0.064 4.486 0.062
0.934 6.39E-06 828.5737 1102 3.197 0.058 3.478 0.061 0.919 1.44E-05
810.5969 1101 3.785 0.100 4.234 0.080 0.894 2.51E-05 809.5936 1101
4.804 0.094 5.194 0.079 0.925 2.81E-05 517.314 1101 5.339 0.065
5.575 0.069 0.958 3.58E-05 495.332 1101 5.003 0.083 5.391 0.112
0.928 0.0001 518.3174 1101 3.325 0.062 3.601 0.073 0.923 0.0001
793.5386 1102 3.632 0.061 3.827 0.039 0.949 0.0001 794.5421 1102
2.536 0.061 2.763 0.040 0.918 0.0001 832.5791 1101 3.305 0.080
3.607 0.061 0.916 0.0001 853.5854 1102 2.527 0.065 2.790 0.052
0.906 0.0001 915.5191 1101 2.278 0.083 2.632 0.063 0.866 0.0001
183.0661 1101 2.324 0.143 2.920 0.094 0.796 0.0002 521.3477 1101
3.406 0.099 3.895 0.127 0.875 0.0002 523.3634 1101 3.221 0.102
3.673 0.103 0.877 0.0002 543.3296 1101 3.579 0.072 3.898 0.087
0.918 0.0002 795.555 1102 2.608 0.069 2.893 0.065 0.902 0.0002
831.5759 1101 4.272 0.080 4.531 0.060 0.943 0.0002 886.5582 1102
2.998 0.049 3.149 0.041 0.952 0.0002 496.3355 1101 3.200 0.086
3.620 0.119 0.884 0.0004 827.5448 1101 3.340 0.105 3.752 0.095
0.890 0.0004 819.5551 1102 2.369 0.062 2.571 0.046 0.922 0.0005
522.3511 1101 1.480 0.111 2.005 0.121 0.738 0.0006 817.5377 1102
2.427 0.060 2.627 0.057 0.924 0.001 545.3453 1101 3.512 0.082 3.776
0.078 0.930 0.0011 575.2728 1101 2.381 0.076 2.653 0.074 0.897
0.0012 700.552 1101 2.150 0.094 2.535 0.102 0.848 0.0012 722.5335
1101 2.437 0.062 2.632 0.056 0.926 0.0014 520.3354 1101 2.075 0.117
2.564 0.130 0.809 0.0016 458.2405 1101 1.664 0.064 1.924 0.077
0.865 0.0017 542.3173 1101 2.268 0.092 2.603 0.089 0.871 0.0017
519.3321 1101 3.807 0.121 4.245 0.133 0.897 0.0019 546.3485 1101
1.931 0.069 2.216 0.089 0.872 0.0021 541.3139 1101 4.077 0.092
4.353 0.096 0.937 0.0025 1019.384 1102 3.095 0.042 2.828 0.037
1.094 0.0026 804.7227 1203 1.612 0.233 2.530 0.220 0.637 0.0027
831.7408 1203 3.839 0.137 4.286 0.130 0.896 0.0028 303.1079 1202
5.575 0.057 5.621 0.047 0.992 0.0032 549.4845 1203 5.480 0.127
5.807 0.110 0.944 0.0033 548.4815 1203 6.917 0.122 7.174 0.103
0.964 0.0034 746.5717 1204 3.115 0.058 3.257 0.061 0.957 0.0046
830.7363 1203 4.694 0.139 5.098 0.135 0.921 0.0046 306.2569 1204
3.015 0.059 3.164 0.067 0.953 0.0058 760.5216 1204 4.156 0.053
4.211 0.039 0.987 0.0058 732.4938 1204 4.274 0.060 4.370 0.057
0.978 0.0059 855.6016 1102 3.619 0.062 3.752 0.065 0.965 0.0059
523.4679 1203 3.249 0.212 3.886 0.165 0.836 0.0065 522.4635 1203
4.717 0.208 5.313 0.176 0.888 0.0066 1227.107 1203 3.235 0.116
3.547 0.100 0.912 0.0074 833.7551 1204 2.396 0.213 3.042 0.161
0.788 0.0075 520.4499 1203 3.898 0.194 4.434 0.148 0.879 0.0076
1228.111 1203 2.587 0.203 3.131 0.105 0.826 0.0077 521.4522 1203
2.319 0.204 2.933 0.158 0.790 0.0083 591.3542 1202 4.110 0.064
4.209 0.058 0.976 0.0085 856.672 1202 2.741 0.042 2.473 0.057 1.108
0.0086 777.553 1202 1.958 0.096 2.309 0.130 0.848 0.009 552.5022
1203 3.097 0.142 3.529 0.147 0.878 0.0092 777.5287 1201 2.781 0.055
2.926 0.072 0.951 0.0092 675.6377 1204 3.806 0.112 4.030 0.066
0.945 0.0098 748.5735 1202 3.883 0.036 3.606 0.050 1.077 0.0101
467.807 1101 1.408 0.134 0.863 0.144 1.631 0.0104 832.7523 1203
3.763 0.160 4.237 0.169 0.888 0.0104 837.718 1204 2.737 0.273 3.548
0.217 0.771 0.0107 745.5658 1204 4.005 0.060 4.109 0.068 0.975
0.0109 568.3573 1202 3.960 0.053 4.037 0.060 0.981 0.0113 550.4957
1203 6.848 0.150 7.184 0.160 0.953 0.0114 833.7571 1203 2.911 0.156
3.397 0.170 0.857 0.0115 838.7226 1204 2.158 0.237 2.880 0.206
0.749 0.0119 551.4986 1203 5.429 0.150 5.801 0.158 0.936 0.0121
585.2673 1202 1.612 0.153 1.034 0.148 1.558 0.0125 821.5712 1102
3.120 0.056 3.200 0.044 0.975 0.0133 835.7006 1204 2.553 0.198
3.113 0.167 0.820 0.0136 731.4916 1204 5.523 0.062 5.567 0.063
0.992 0.0138 867.7579 1204 2.503 0.252 3.205 0.189 0.781 0.0139
482.3215 1202 2.040 0.058 2.181 0.062 0.935 0.0145 858.6861 1202
2.935 0.050 2.670 0.052 1.099 0.0145 584.2646 1204 3.282 0.185
2.567 0.172 1.279 0.0148 829.7246 1203 4.743 0.119 4.973 0.101
0.954 0.015 828.7207 1203 5.497 0.121 5.716 0.107 0.962 0.0154
723.5197 1204 4.571 0.070 4.214 0.056 1.085 0.0167 864.738 1204
4.725 0.142 5.051 0.141 0.936 0.0171 691.1957 1102 2.168 0.067
1.878 0.068 1.155 0.0181 585.2664 1204 1.716 0.183 1.077 0.170
1.593 0.0187 749.5777 1202 2.879 0.034 2.659 0.046 1.083 0.0189
757.5014 1204 3.770 0.066 3.847 0.052 0.980 0.0197 863.7336 1204
5.458 0.146 5.763 0.147 0.947 0.02 830.7352 1204 4.133 0.151 4.493
0.150 0.920 0.0202 252.2095 1204 1.665 0.076 1.852 0.070 0.899
0.0206 860.7756 1203 4.306 0.109 4.507 0.100 0.955 0.0222 863.6876
1204 4.889 0.090 5.026 0.092 0.973 0.0227 861.7806 1203 2.435 0.158
2.811 0.119 0.866 0.0228 840.6063 1202 2.927 0.050 2.664 0.059
1.099 0.0229 632.5762 1203 1.325 0.170 1.773 0.132 0.747 0.0231
826.7069 1204 2.424 0.137 2.731 0.099 0.887 0.0242 419.8773 1101
1.946 0.176 1.305 0.194 1.491 0.0243 440.3532 1204 1.481 0.134
0.945 0.180 1.567 0.0247 831.7387 1204 3.137 0.164 3.534 0.150
0.888 0.0248 310.2883 1204 2.448 0.085 2.607 0.062 0.939 0.0256
481.3172 1202 3.978 0.053 4.029 0.060 0.987 0.0259 205.8865 1101
3.759 0.234 2.793 0.319 1.346 0.0271 513.4122 1204 1.142 0.153
0.623 0.163 1.833 0.0279 839.6031 1202 3.895 0.051 3.609 0.057
1.079 0.0286 865.7487 1204 4.847 0.188 5.275 0.193 0.919 0.029
579.5325 1203 4.188 0.154 4.491 0.127 0.933 0.0292 807.59 1202
2.477 0.072 2.633 0.082 0.941 0.0293 351.8903 1101 2.359 0.220
1.567 0.256 1.506 0.0296 782.5085 1204 3.596 0.062 3.662 0.055
0.982 0.0297 859.7696 1203 5.853 0.104 5.972 0.093 0.980 0.0298
409.8484 1101 2.107 0.192 1.437 0.210 1.466 0.0304 577.5137 1203
8.099 0.104 8.148 0.094 0.994 0.0305 1018.942 1203 3.468 0.142
3.767 0.126 0.921 0.0305 703.5709 1101 3.024 0.090 3.185 0.079
0.949 0.0314 484.3794 1204 2.234 0.118 1.721 0.178 1.298 0.0315
283.2602 1204 6.622 0.070 6.621 0.067 1.000 0.0316 401.8166 1101
1.696 0.161 1.139 0.177 1.489 0.0316 832.7492 1204 3.251 0.195
3.728 0.190 0.872 0.0316 190.033 1101 3.264 0.233 2.384 0.289 1.369
0.0317 806.5873 1202 4.367 0.082 4.490 0.092 0.973 0.032 282.2573
1204 9.088 0.073 9.019 0.071 1.008 0.0327 195.8577 1101 3.161 0.202
2.370 0.266 1.334 0.0328 744.4956 1204 3.941 0.058 3.988 0.058
0.988 0.0332 866.7532 1204 4.123 0.184 4.547 0.189 0.907 0.0334
215.9153 1101 5.221 0.247 4.183 0.346 1.248 0.0335 825.6926 1203
1.767 0.173 2.165 0.124 0.816 0.0339 469.8042 1101 1.617 0.150
1.096 0.170 1.475 0.0344 874.7645 1204 3.091 0.144 3.377 0.111
0.915 0.0345 494.4343 1203 2.119 0.275 2.809 0.232 0.754 0.0346
399.8196 1101 1.827 0.171 1.249 0.184 1.463 0.0348 802.7056 1204
1.701 0.206 2.248 0.202 0.756 0.0348 576.5098 1203 9.446 0.111
9.460 0.099 0.999 0.0355 382.1082 1101 2.222 0.229 1.494 0.220
1.487 0.0356 720.6438 1204 2.962 0.180 3.293 0.083 0.900 0.036
805.5839 1202 5.574 0.081 5.658 0.097 0.985 0.0365 779.5441 1201
7.096 0.041 7.010 0.053 1.012 0.0368 231.8893 1101 2.592 0.233
1.800 0.259 1.440 0.0372 686.4877 1204 2.721 0.054 2.791 0.052
0.975 0.0374 429.3749 1204 2.601 0.098 2.777 0.077 0.937 0.0378
265.8423 1101 2.499 0.220 1.743 0.252 1.433 0.0384 722.6424 1204
3.726 0.167 4.006 0.086 0.930 0.0385 780.5474 1201 6.022 0.043
5.972 0.054 1.008 0.0396 599.4936 1204 1.373 0.191 1.873 0.188
0.733 0.04 536.4794 1203 2.280 0.202 2.711 0.143 0.841 0.0422
244.0559 1101 3.574 0.126 3.098 0.132 1.153 0.0424 202.0453 1101
5.561 0.180 4.832 0.215 1.151 0.0425 535.7944 1101 1.480 0.143
1.018 0.153 1.455 0.0425 758.5092 1204 4.670 0.051 4.658 0.042
1.003 0.0426 626.5104 1204 2.100 0.135 2.391 0.114 0.878 0.0427
847.5316 1101 2.815 0.104 2.994 0.083 0.940 0.0432 592.3571 1202
2.375 0.065 2.480 0.062 0.958 0.0437 569.4784 1204 1.175 0.183
0.634 0.178 1.853 0.0447 721.639 1204 4.879 0.142 5.064 0.088 0.963
0.0447 752.5583 1204 4.284 0.070 3.966 0.061 1.080 0.0451 507.7055
1101 1.487 0.126 1.052 0.154 1.413 0.0454 827.7086 1203 3.916 0.119
4.098 0.096 0.956 0.046 784.5237 1204 3.464 0.056 3.490 0.037 0.993
0.0461 755.486 1204 3.395 0.059 3.450 0.057 0.984 0.0471 628.5237
1204 2.627 0.158 2.955 0.136 0.889 0.0472 218.0192 1101 3.591 0.258
2.716 0.302 1.322 0.049
TABLE-US-00006 TABLE 6 Accurate mass features differing between
clinically diagnosed AD patients with a mild cognitive impairment
and non-demented controls (p < 0.05, log2 transformed). AVG AVG
Detected Analysis (log2) SEM (log2) SEM Mass Mode AD noCog AD noCog
Normal Normal log(2) Ratio P Value 723.5197 1204 3.576 0.058 4.350
0.056 0.822 6.42E-14 723.5195 1202 2.181 0.056 2.892 0.052 0.754
2.49E-13 751.5529 1202 3.293 0.050 3.920 0.050 0.840 1.39E-12
724.5257 1204 2.883 0.044 3.463 0.055 0.833 9.96E-12 749.5367 1202
3.168 0.051 3.714 0.041 0.853 1.33E-11 752.5564 1202 2.216 0.057
2.836 0.050 0.781 1.91E-11 751.5555 1204 4.553 0.060 5.248 0.060
0.868 1.96E-11 750.5402 1202 2.175 0.054 2.704 0.044 0.804 2.92E-10
752.5583 1204 3.453 0.063 4.094 0.061 0.844 6.11E-10 750.544 1204
3.299 0.052 3.858 0.063 0.855 3.31E-09 749.5407 1204 4.429 0.061
5.012 0.064 0.884 9.50E-09 699.5198 1204 2.309 0.047 2.775 0.054
0.832 1.20E-08 725.5385 1204 2.958 0.060 3.417 0.054 0.866 4.59E-07
727.5568 1204 3.588 0.050 3.986 0.060 0.900 3.00E-06 812.5762 1202
1.571 0.087 2.084 0.058 0.754 1.13E-05 541.3432 1102 3.518 0.039
3.798 0.048 0.926 2.42E-05 813.5885 1202 2.992 0.041 3.276 0.048
0.913 2.68E-05 811.5732 1202 2.683 0.055 3.027 0.055 0.886 3.92E-05
726.5461 1204 2.869 0.047 3.150 0.050 0.911 0.0001 814.5917 1202
2.028 0.040 2.289 0.051 0.886 0.0001 632.5762 1203 0.944 0.175
1.830 0.132 0.516 0.0002 728.5627 1204 2.998 0.045 3.281 0.060
0.914 0.0003 782.5085 1204 3.435 0.071 3.780 0.055 0.909 0.0003
569.3687 1102 2.479 0.045 2.724 0.048 0.910 0.0004 803.5445 1101
5.324 0.070 5.655 0.059 0.941 0.0007 804.5476 1101 4.370 0.073
4.703 0.060 0.929 0.0009 817.5377 1102 2.405 0.066 2.712 0.057
0.887 0.001 787.5729 1202 1.864 0.058 2.145 0.057 0.869 0.0011
747.5245 1204 3.521 0.063 3.886 0.090 0.906 0.0013 775.5533 1202
2.140 0.062 2.449 0.068 0.874 0.0013 746.5717 1204 3.117 0.043
3.362 0.061 0.927 0.0014 827.57 1102 4.337 0.063 4.630 0.062 0.937
0.0017 745.5658 1204 3.989 0.041 4.242 0.068 0.940 0.0018 828.5737
1102 3.313 0.063 3.590 0.061 0.923 0.0025 542.3173 1101 2.344 0.067
2.687 0.089 0.872 0.0027 760.5216 1204 4.140 0.052 4.347 0.039
0.952 0.0028 748.5287 1204 2.319 0.131 2.820 0.088 0.822 0.003
774.0316 1204 2.044 0.199 1.167 0.203 1.752 0.0031 784.5237 1204
3.405 0.052 3.603 0.037 0.945 0.0035 775.5528 1204 3.221 0.065
3.537 0.085 0.911 0.004 744.5536 1204 4.388 0.046 4.605 0.058 0.953
0.0042 482.3215 1202 2.001 0.057 2.251 0.062 0.889 0.0043 768.5539
1204 3.995 0.060 4.279 0.077 0.934 0.0044 506.2851 1201 3.210 0.100
2.656 0.165 1.209 0.0045 747.5201 1202 2.006 0.060 2.313 0.088
0.867 0.0045 755.486 1204 3.295 0.069 3.561 0.057 0.925 0.0047
781.5617 1101 6.305 0.075 6.610 0.072 0.954 0.005 786.5416 1204
3.903 0.046 4.087 0.043 0.955 0.0052 804.5713 1102 4.426 0.054
4.610 0.033 0.960 0.0064 743.5471 1204 5.349 0.054 5.579 0.063
0.959 0.0066 541.3139 1101 4.172 0.067 4.494 0.096 0.928 0.0069
767.5495 1204 5.085 0.063 5.369 0.082 0.947 0.0072 782.565 1101
5.279 0.080 5.589 0.078 0.945 0.0074 555.3102 1102 2.033 0.059
2.240 0.046 0.908 0.0086 567.3547 1102 3.031 0.045 3.218 0.054
0.942 0.0089 565.3394 1102 3.728 0.064 3.958 0.055 0.942 0.0094
582.2473 1201 3.346 0.151 2.793 0.141 1.198 0.0099 803.568 1102
5.656 0.057 5.838 0.035 0.969 0.0108 779.5444 1101 6.654 0.070
6.874 0.043 0.968 0.0113 796.5876 1204 2.630 0.063 2.860 0.062
0.919 0.0117 758.5092 1204 4.652 0.043 4.808 0.042 0.968 0.0126
783.5148 1204 3.344 0.068 3.574 0.056 0.936 0.0126 748.5735 1202
3.893 0.045 3.722 0.050 1.046 0.0135 529.3167 1202 3.028 0.064
3.265 0.069 0.927 0.0138 781.562 1201 7.406 0.060 7.638 0.071 0.970
0.0144 780.5474 1101 5.661 0.070 5.875 0.043 0.964 0.0145 810.5399
1204 2.905 0.069 3.145 0.066 0.923 0.0147 837.5881 1202 2.412 0.040
2.577 0.053 0.936 0.0147 590.343 1202 4.042 0.072 4.304 0.076 0.939
0.0149 580.5351 1203 1.531 0.200 2.192 0.174 0.699 0.0166 789.5892
1202 1.755 0.049 1.952 0.065 0.899 0.0167 887.7352 1204 6.359 0.115
5.963 0.113 1.066 0.0174 828.5743 1202 5.301 0.087 5.598 0.085
0.947 0.0178 916.7743 1204 5.952 0.113 5.571 0.107 1.068 0.0178
856.672 1202 2.753 0.058 2.553 0.057 1.078 0.0179 827.5701 1202
6.362 0.095 6.684 0.092 0.952 0.0187 942.7879 1204 3.531 0.102
3.195 0.092 1.105 0.019 523.4679 1203 3.471 0.153 4.011 0.165 0.865
0.0194 915.7681 1204 6.365 0.115 5.972 0.116 1.066 0.0194 517.314
1101 5.547 0.055 5.755 0.069 0.964 0.0195 777.553 1202 2.002 0.098
2.384 0.130 0.840 0.0204 744.4956 1204 3.916 0.061 4.117 0.058
0.951 0.0206 440.3532 1204 1.502 0.136 0.975 0.180 1.539 0.021
782.5653 1201 6.382 0.065 6.614 0.076 0.965 0.0222 494.4343 1203
2.145 0.226 2.900 0.232 0.740 0.0232 303.1079 1202 5.646 0.048
5.802 0.047 0.973 0.0233 771.5814 1204 4.208 0.050 4.367 0.046
0.964 0.0236 521.4522 1203 2.482 0.173 3.028 0.158 0.820 0.0242
793.5681 1204 3.191 0.055 3.372 0.055 0.946 0.0244 832.6026 1102
3.662 0.042 3.795 0.040 0.965 0.0252 589.3403 1202 5.900 0.084
6.171 0.085 0.956 0.026 732.4938 1204 4.319 0.061 4.511 0.057 0.957
0.0263 783.5672 1101 3.993 0.085 4.259 0.079 0.938 0.0265 569.369
1202 4.947 0.042 5.089 0.047 0.972 0.0273 522.4635 1203 4.938 0.167
5.485 0.176 0.900 0.0277 795.5181 1101 2.823 0.073 3.044 0.064
0.927 0.0287 888.7394 1204 5.759 0.115 5.400 0.110 1.066 0.0287
490.3641 1203 1.864 0.138 1.397 0.159 1.334 0.029 914.7583 1204
5.125 0.112 4.763 0.118 1.076 0.0294 520.4499 1203 4.141 0.131
4.577 0.148 0.905 0.03 805.5832 1102 4.292 0.055 4.485 0.068 0.957
0.0301 795.5838 1204 3.558 0.070 3.769 0.063 0.944 0.0305 575.2728
1101 2.532 0.060 2.739 0.074 0.925 0.0324 831.5997 1102 4.774 0.042
4.903 0.040 0.974 0.0325 518.3174 1101 3.523 0.054 3.717 0.073
0.948 0.0338 520.3354 1101 2.271 0.117 2.646 0.130 0.858 0.0354
573.4852 1203 4.919 0.063 4.740 0.053 1.038 0.0354 579.5325 1203
4.235 0.135 4.636 0.127 0.913 0.0355 570.3725 1202 3.021 0.043
3.155 0.045 0.958 0.0359 793.5386 1102 3.799 0.057 3.950 0.039
0.962 0.0359 481.3172 1202 3.980 0.058 4.159 0.060 0.957 0.0361
831.5759 1101 4.485 0.066 4.677 0.060 0.959 0.0365 913.7513 1204
5.358 0.113 5.006 0.120 1.070 0.037 772.5862 1204 3.306 0.045 3.437
0.042 0.962 0.0378 304.241 1204 4.898 0.062 5.088 0.066 0.963
0.0397 746.557 1202 2.177 0.042 2.057 0.037 1.058 0.0399 519.3321
1101 4.014 0.116 4.382 0.133 0.916 0.0405 833.7551 1204 2.611 0.191
3.140 0.161 0.831 0.0406 889.7492 1204 7.711 0.122 7.365 0.110
1.047 0.0415 614.4914 1203 2.747 0.091 2.448 0.114 1.122 0.0418
618.4829 1201 1.844 0.179 1.310 0.185 1.407 0.0429 601.5164 1203
7.639 0.052 7.463 0.069 1.024 0.0431 671.5723 1204 2.349 0.088
2.604 0.087 0.902 0.0435 794.5421 1102 2.703 0.059 2.853 0.040
0.948 0.0438 731.4916 1204 5.555 0.068 5.746 0.063 0.967 0.0444
512.4082 1204 2.523 0.180 1.954 0.215 1.292 0.0451 768.5503 1202
2.028 0.082 2.277 0.090 0.891 0.0453 665.501 1204 3.429 0.105 3.074
0.142 1.116 0.0454 890.7535 1204 7.055 0.116 6.731 0.106 1.048
0.0454 759.5163 1204 4.804 0.055 4.954 0.048 0.970 0.0455 605.5457
1203 5.235 0.100 5.509 0.089 0.950 0.0473 541.4425 1204 2.234 0.148
1.745 0.198 1.280 0.0488 572.4816 1203 6.271 0.061 6.108 0.052
1.027 0.049 743.5466 1203 1.977 0.103 2.266 0.099 0.873 0.0492
TABLE-US-00007 TABLE 7 Accurate mass features differing between
dementia patients with a significant cognitive impairment (ADAS
.gtoreq. 16) and dementia patients with a mild cognitive impairment
(ADAS .ltoreq. 15) (p < 0.05, log2 transformed). Detected
Analysis AVG (log2) SEM AVG (log2) SEM log(2) Mass Mode ADAS high
ADAS HIGH ADAS LOW ADAS LOW Ratio P Value 541.3432 1102 3.155 0.038
3.581 0.054 0.881 6.71E-09 567.3547 1102 2.633 0.052 3.122 0.058
0.843 3.84E-08 569.3687 1102 2.079 0.053 2.526 0.054 0.823 2.41E-07
829.5856 1102 3.861 0.057 4.303 0.059 0.897 1.71E-06 565.3394 1102
3.268 0.064 3.784 0.081 0.864 3.73E-06 837.5027 1101 3.373 0.056
3.818 0.070 0.884 4.53E-06 804.5713 1102 4.022 0.051 4.414 0.060
0.911 5.01E-06 831.5997 1102 4.379 0.055 4.761 0.046 0.920 5.17E-06
832.6026 1102 3.278 0.055 3.658 0.046 0.896 5.66E-06 803.568 1102
5.252 0.052 5.640 0.065 0.931 1.42E-05 805.5832 1102 3.894 0.056
4.293 0.063 0.907 1.48E-05 871.5528 1102 2.886 0.051 3.244 0.059
0.890 2.25E-05 555.3102 1102 1.669 0.053 2.070 0.074 0.806 2.34E-05
853.5854 1102 2.473 0.058 2.824 0.040 0.876 2.85E-05 886.5582 1102
2.898 0.036 3.166 0.051 0.916 4.34E-05 808.5792 1101 4.475 0.071
4.922 0.068 0.909 4.77E-05 780.5474 1101 5.229 0.067 5.665 0.086
0.923 0.0001 805.5605 1101 5.210 0.072 5.655 0.078 0.921 0.0001
807.5758 1101 5.522 0.071 5.962 0.069 0.926 0.0001 543.3296 1101
3.594 0.055 3.948 0.075 0.910 0.0002 779.5444 1101 6.228 0.067
6.660 0.086 0.935 0.0002 806.5639 1101 4.213 0.074 4.656 0.079
0.905 0.0002 829.5604 1101 3.368 0.070 3.766 0.069 0.894 0.0002
755.5468 1101 2.034 0.091 2.551 0.098 0.797 0.0003 819.5551 1102
2.270 0.062 2.611 0.062 0.869 0.0003 915.5191 1101 2.209 0.066
2.603 0.082 0.849 0.0004 794.5421 1102 2.389 0.054 2.705 0.073
0.883 0.0007 832.5791 1101 3.221 0.066 3.564 0.066 0.904 0.0007
793.5386 1102 3.462 0.057 3.778 0.069 0.916 0.0008 803.5445 1101
4.995 0.065 5.351 0.078 0.933 0.0008 804.5476 1101 4.045 0.064
4.402 0.080 0.919 0.0008 731.5464 1101 1.574 0.132 2.200 0.109
0.716 0.0011 827.57 1102 4.055 0.057 4.341 0.062 0.934 0.0014
722.5335 1101 2.379 0.064 2.687 0.064 0.885 0.0015 831.5759 1101
4.163 0.072 4.510 0.070 0.923 0.0015 517.314 1101 5.343 0.057 5.631
0.068 0.949 0.0019 757.4991 1101 3.393 0.086 3.828 0.102 0.886
0.0019 783.5672 1101 3.547 0.087 3.974 0.097 0.893 0.002 545.3453
1101 3.457 0.072 3.795 0.073 0.911 0.0022 518.3174 1101 3.321 0.056
3.600 0.068 0.922 0.0023 760.5811 1101 4.181 0.092 4.609 0.101
0.907 0.003 784.5811 1101 4.155 0.104 4.630 0.111 0.897 0.0033
546.3485 1101 1.883 0.067 2.196 0.078 0.857 0.0034 759.5779 1101
5.313 0.091 5.727 0.098 0.928 0.0034 855.6016 1102 3.447 0.060
3.709 0.056 0.929 0.0034 847.5316 1101 2.627 0.075 3.004 0.106
0.874 0.0037 781.5617 1101 5.942 0.079 6.291 0.082 0.945 0.0039
575.2728 1101 2.359 0.070 2.663 0.069 0.886 0.0042 828.5737 1102
3.070 0.054 3.317 0.064 0.926 0.0047 795.555 1102 2.558 0.065 2.831
0.063 0.904 0.0053 821.5712 1102 2.952 0.058 3.195 0.057 0.924
0.0053 833.5932 1101 3.069 0.092 3.444 0.083 0.891 0.0053 782.565
1101 4.918 0.082 5.266 0.085 0.934 0.0056 795.5181 1101 2.462 0.075
2.793 0.090 0.882 0.0066 725.5527 1101 3.222 0.073 3.500 0.057
0.921 0.0068 761.5843 1101 2.194 0.095 2.580 0.098 0.850 0.0078
738.5448 1102 2.493 0.076 2.766 0.047 0.901 0.0083 811.6096 1101
2.869 0.106 3.277 0.096 0.876 0.0088 758.5656 1101 5.459 0.101
5.870 0.112 0.930 0.0092 785.5933 1101 5.643 0.108 6.049 0.097
0.933 0.0103 458.2405 1101 1.635 0.057 1.880 0.075 0.870 0.0104
757.5626 1101 6.418 0.100 6.815 0.112 0.942 0.0112 541.3139 1101
3.968 0.074 4.278 0.096 0.928 0.0116 786.5967 1101 4.570 0.109
4.973 0.099 0.919 0.012 748.5721 1102 3.811 0.069 4.050 0.055 0.941
0.0137 749.5761 1102 2.699 0.066 2.929 0.053 0.922 0.0156 501.3212
1201 1.903 0.052 2.108 0.066 0.903 0.0176 809.5936 1101 4.792 0.099
5.124 0.084 0.935 0.0194 810.5969 1101 3.785 0.102 4.126 0.086
0.917 0.0199 542.3173 1101 2.183 0.077 2.464 0.091 0.886 0.0228
724.5493 1101 4.493 0.076 4.734 0.059 0.949 0.0232 723.5197 1204
3.966 0.083 3.699 0.091 1.072 0.0362 521.3477 1101 3.545 0.086
3.863 0.130 0.918 0.0374 1098.974 1204 3.016 0.105 2.552 0.219
1.182 0.039 523.3634 1101 3.263 0.100 3.596 0.130 0.907 0.0433
807.5768 1201 6.721 0.033 6.816 0.031 0.986 0.0452 826.7069 1204
2.385 0.134 2.766 0.113 0.862 0.0462 931.7695 1203 2.852 0.090
2.495 0.169 1.143 0.0475 183.0661 1101 2.358 0.139 2.748 0.116
0.858 0.0485 504.3814 1203 1.818 0.065 1.573 0.113 1.156 0.0489
TABLE-US-00008 TABLE 8 Accurate mass features differing between
patients with mild cognitive impairment (MMSE 18-23), severe
cognitive impairment (MMSE .ltoreq. 17) and normal cognitive
ability (MMSE .gtoreq. 28) as measured on the MMSE. AVG SEM AVG SEM
log (2) log(2) log(2) (log2) (log2) (log2) (log2) AVG SEM Ratio
Ratio Ratio Detected Analysis mild mild severe severe (log2) (log2)
mild/ severe/ mild/ Mass Mode MMSE MMSE MMSE MMSE Normal Normal
normal normal severe P Value 726.5461 1204 3.125 0.058 2.729 0.053
2.981 0.067 1.048 0.916 1.145 0.0002 675.6377 1204 4.131 0.049
3.856 0.082 4.124 0.061 1.002 0.935 1.071 0.0102 801.555* 1202
2.733 0.144 2.602 0.110 3.052 0.077 0.895 0.853 1.050 0.0108
570.3725 1202 3.071 0.067 2.895 0.054 3.130 0.054 0.981 0.925 1.061
0.0116 597.484 1203 2.915 0.146 2.582 0.101 3.046 0.110 0.957 0.848
1.129 0.0138 246.1472 1204 2.437 0.187 3.031 0.161 2.974 0.067
0.819 1.019 0.804 0.0161 565.3394* 1202 7.087 0.132 6.908 0.090
7.304 0.087 0.970 0.946 1.026 0.0171 596.4791 1203 4.125 0.172
3.792 0.112 4.283 0.111 0.963 0.886 1.088 0.0188 246.1465 1202
3.577 0.208 4.046 0.123 4.169 0.104 0.858 0.970 0.884 0.0199
432.3254 1204 2.540 0.059 2.325 0.058 2.495 0.047 1.018 0.932 1.092
0.0201 569.369* 1202 5.009 0.056 4.834 0.050 5.032 0.063 0.995
0.961 1.036 0.0285 661.6233 1204 2.783 0.091 2.448 0.090 2.683
0.079 1.037 0.912 1.137 0.0307 857.6186* 1202 3.923 0.122 3.699
0.104 4.090 0.101 0.959 0.904 1.061 0.0332 804.5718 1202 5.940
0.135 5.776 0.088 6.170 0.112 0.963 0.936 1.028 0.0334 803.5677
1202 7.274 0.174 7.084 0.116 7.570 0.131 0.961 0.936 1.027 0.0359
542.3461 1202 3.898 0.134 3.773 0.071 4.124 0.104 0.945 0.915 1.033
0.037 810.5399 1204 3.152 0.104 2.749 0.088 2.959 0.113 1.065 0.929
1.146 0.0374 728.5627 1204 3.233 0.078 2.954 0.065 3.054 0.071
1.059 0.967 1.094 0.0404 566.3434 1202 5.338 0.100 5.142 0.093
5.460 0.082 0.978 0.942 1.038 0.0417 481.3172 1202 3.940 0.088
3.875 0.069 4.136 0.077 0.953 0.937 1.017 0.0436 724.5257 1204
3.168 0.102 2.875 0.079 2.961 0.055 1.070 0.971 1.102 0.0447
825.5544 1202 2.933 0.173 2.605 0.144 3.160 0.171 0.928 0.824 1.126
0.0474 826.5581 1202 2.005 0.151 1.579 0.156 2.139 0.188 0.937
0.738 1.270 0.0488
TABLE-US-00009 TABLE 9 Grouping of patients into one of 8 groups
based on the presence of AD pathology, ADAS score and MMSE score. A
score of 1 was given for the presence of AD pathology, high ADAS
score (.gtoreq.16), or low MMSE score (.ltoreq.23); a score of 0
was given in the absence of AD pathology, low ADAS score
(.ltoreq.15), or high MMSE score (.gtoreq.28). AD Patient ID
Pathology ADAS MMSE 7001 1 1 1 7002 0 1 1 7006 1 0 1 7007 1 1 1
7008 1 0 1 7009 1 0 1 7010 1 0 1 7011 1 0 1 7014 1 0 1 7015 0 1 1
7016 1 1 1 7017 1 1 1 7022 1 1 1 7023 1 1 1 7024 1 0 1 7025 1 0 1
7027 1 0 1 7028 1 1 1 7029 1 1 1 7030 1 0 1 7031 1 0 1 7033 0 1 1
7034 1 0 1 7035 0 1 1 7037 1 1 1 7039 1 0 1 7042 1 0 1 7043 0 1 1
7044 1 1 1 7045 1 0 1 7046 1 1 1 7047 0 1 1 7048 1 0 1 7049 1 0 1
7050 0 1 1 7051 1 0 1 7052 1 0 1 7053 1 0 1 7055 0 1 1 7056 1 1 1
7057 1 0 1 7058 1 0 1 7059 0 1 1 7062 0 0 0 7063 1 1 1 7064 0 0 0
7066 0 0 1 7067 1 1 1 7070 0 0 0 7074 0 1 1 7075 1 1 1 7076 0 1 1
7077 1 1 1 7078 1 0 1 7079 0 1 1 7080 0 1 1 7081 1 1 1 7082 1 1 1
7084 1 1 1 7085 1 0 1 7086 0 1 1 7087 0 1 1 7088 1 1 1 7089 1 1 1
7090 1 1 1 7091 0 1 1 7093 1 0 1 7094 0 1 1 7095 0 1 1 7096 1 0 1
7097 1 0 1 7098 0 0 0 7101 0 0 0 7102 1 0 0 7103 0 1 0 7104 0 1 0
7105 0 1 0 7106 1 1 0 7108 0 0 0 7109 0 0 0 7110 0 1 0 7111 0 1 0
7112 1 1 0 7113 1 1 0 7114 1 1 0 7115 0 1 0 7116 1 1 0 7117 0 1 0
7118 0 0 0 7119 0 0 0 7120 0 0 0 7124 1 1 0 7125 1 1 0 7126 0 0 0
7127 0 0 0 7128 0 1 0 7129 1 0 0 7130 0 0 0 7131 0 0 0 7133 0 1 0
7134 0 1 0 7135 0 0 0 7136 1 0 0 7137 0 0 0 7138 0 0 0 7139 0 1 0
7140 1 0 0 7141 1 1 0 7142 1 1 0 7143 1 0 0 7144 1 0 0 7145 0 1 0
7146 0 1 0 7147 0 1 0 7150 1 0 0 7151 1 0 0 7152 0 1 0 7153 0 0 0
7154 1 1 0 7155 1 1 0 7156 0 0 0 7157 1 0 0 7158 1 0 0 7309 1 0 1
7310 1 0 0 7315 1 0 1 7802 0 0 0 7811 0 0 0 7814 1 0 0 7817 1 0 0
7818 0 0 0 7819 0 1 0 7823 1 0 0 7831 0 0 0 7832 1 0 0 7833 1 0
0
TABLE-US-00010 TABLE 10 Accurate mass features differing between
patients showing the best discrimination between AD and non-AD
pathology (p < 0.05, log2 transformed). AVG AVG Detected
Analysis (log2) (log2) log(2) Mass Mode AD SEM AD non-AD SEM AD
Ratio P Value 699.5199 1204 1.985 0.043 2.956 0.067 0.672 1.24E-14
723.5195 1204 3.281 0.070 4.702 0.101 0.698 4.86E-14 723.5194 1202
2.039 0.077 3.191 0.069 0.639 1.59E-13 751.5553 1204 4.290 0.074
5.450 0.093 0.787 5.86E-12 724.5258 1204 2.667 0.062 3.750 0.092
0.711 6.50E-12 751.5529 1202 3.197 0.068 4.076 0.062 0.784 1.03E-11
752.5564 1202 2.126 0.069 3.010 0.061 0.706 1.16E-11 749.5367 1202
3.061 0.062 3.861 0.056 0.793 1.22E-11 752.5581 1204 3.170 0.072
4.292 0.094 0.739 1.52E-11 749.5406 1204 4.177 0.067 5.235 0.095
0.798 4.77E-11 750.5403 1202 2.081 0.074 2.886 0.051 0.721 6.77E-11
750.5438 1204 3.060 0.068 4.067 0.091 0.752 7.99E-11 727.5567 1204
3.305 0.074 4.064 0.053 0.813 4.71E-10 725.5385 1204 2.654 0.094
3.590 0.079 0.739 3.99E-09 728.5629 1204 2.813 0.063 3.345 0.057
0.841 2.54E-07 726.5462 1204 2.753 0.064 3.215 0.062 0.856 7.09E-06
768.5538 1204 3.755 0.060 4.250 0.084 0.884 2.34E-05 747.5244 1204
3.263 0.092 3.999 0.123 0.816 2.45E-05 744.5535 1204 4.147 0.050
4.492 0.053 0.923 3.13E-05 813.5885 1202 2.969 0.051 3.362 0.067
0.883 3.58E-05 743.547 1204 5.096 0.053 5.455 0.059 0.934 0.0001
767.5494 1204 4.835 0.065 5.337 0.092 0.906 0.0001 775.5527 1204
2.993 0.100 3.645 0.105 0.821 0.0001 775.5533 1202 1.993 0.078
2.525 0.088 0.789 0.0001 811.5732 1202 2.590 0.072 3.025 0.071
0.856 0.0001 812.5761 1202 1.599 0.064 2.057 0.084 0.777 0.0001
817.5376 1102 1.992 0.073 2.490 0.081 0.800 0.0001 747.5201 1202
1.829 0.090 2.412 0.110 0.758 0.0002 795.5837 1204 3.418 0.073
3.810 0.062 0.897 0.0002 796.5874 1204 2.468 0.051 2.839 0.074
0.869 0.0002 748.5286 1204 1.812 0.247 2.917 0.126 0.621 0.0003
810.54 1204 2.627 0.088 3.116 0.083 0.843 0.0003 782.5084 1204
3.308 0.060 3.687 0.078 0.897 0.0004 755.486 1204 3.069 0.080 3.477
0.073 0.883 0.0005 758.5092 1204 4.418 0.065 4.707 0.043 0.939
0.0007 771.5813 1204 3.965 0.074 4.288 0.046 0.925 0.0007 304.241
1204 4.716 0.057 5.013 0.062 0.941 0.0011 814.5919 1202 1.988 0.046
2.307 0.077 0.862 0.0011 786.5415 1204 3.671 0.074 3.979 0.050
0.923 0.0013 787.5728 1202 1.743 0.085 2.136 0.075 0.816 0.0013
793.5386 1102 3.337 0.086 3.690 0.057 0.904 0.0015 783.5147 1204
3.166 0.071 3.512 0.077 0.901 0.0022 828.5736 1102 2.906 0.080
3.292 0.087 0.883 0.0023 828.5743 1202 5.132 0.073 5.485 0.081
0.936 0.0024 743.5468 1203 1.812 0.118 2.235 0.061 0.811 0.0029
784.5235 1204 3.196 0.081 3.490 0.045 0.916 0.003 760.5215 1204
3.910 0.072 4.200 0.058 0.931 0.0032 787.5464 1204 2.617 0.090
2.937 0.050 0.891 0.0037 827.57 1102 3.933 0.078 4.313 0.096 0.912
0.0039 305.2438 1204 2.384 0.064 2.648 0.058 0.900 0.0041 794.5421
1102 2.275 0.090 2.601 0.058 0.875 0.0041 827.5701 1202 6.193 0.081
6.559 0.090 0.944 0.0046 590.343 1202 3.939 0.055 4.216 0.074 0.934
0.0047 589.3404 1202 5.769 0.061 6.076 0.085 0.950 0.0057 789.5893
1202 1.673 0.064 1.967 0.077 0.851 0.0057 670.5689 1204 3.048 0.095
3.510 0.127 0.868 0.006 855.6016 1102 3.325 0.079 3.646 0.079 0.912
0.0064 759.5162 1204 4.510 0.086 4.811 0.060 0.937 0.0065 671.5722
1204 2.157 0.123 2.635 0.111 0.819 0.0066 769.5654 1204 3.829 0.065
4.086 0.063 0.937 0.0069 856.6063 1202 4.446 0.076 4.777 0.087
0.931 0.007 409.0208 1202 2.638 0.057 2.899 0.073 0.910 0.0072
774.0313 1204 2.317 0.202 1.370 0.268 1.691 0.0076 804.5713 1102
3.964 0.086 4.276 0.071 0.927 0.008 855.6025 1202 5.437 0.074 5.749
0.085 0.946 0.0083 530.382 1204 2.991 0.089 2.097 0.309 1.426
0.0085 626.5278 1203 3.676 0.068 3.915 0.053 0.939 0.0087 731.4914
1204 5.281 0.086 5.589 0.073 0.945 0.0093 732.4937 1204 4.050 0.083
4.348 0.070 0.931 0.0093 517.3722 1204 2.189 0.112 1.465 0.241
1.494 0.0096 741.5307 1204 2.754 0.086 3.117 0.105 0.884 0.0109
803.5681 1102 5.187 0.087 5.491 0.073 0.945 0.0111 746.5714 1204
2.962 0.065 3.228 0.076 0.918 0.0113 627.5304 1203 2.544 0.068
2.783 0.058 0.914 0.0118 544.397 1204 3.249 0.120 2.323 0.341 1.399
0.0145 745.5656 1204 3.812 0.071 4.091 0.083 0.932 0.0148 739.5142
1204 2.576 0.105 3.009 0.137 0.856 0.0162 570.3725 1202 2.879 0.050
3.071 0.057 0.938 0.0164 686.4877 1204 2.527 0.068 2.757 0.064
0.917 0.0179 517.3136 1201 2.907 0.104 2.575 0.088 1.129 0.0193
744.4956 1204 3.741 0.073 3.988 0.070 0.938 0.0202 529.3167 1202
2.929 0.057 3.199 0.095 0.916 0.0203 821.5713 1102 2.825 0.089
3.091 0.065 0.914 0.0207 819.5641 1202 2.632 0.117 3.063 0.136
0.859 0.0209 667.5474 1204 3.336 0.118 2.882 0.150 1.158 0.0222
886.5582 1102 2.820 0.064 3.038 0.067 0.928 0.0232 796.529 1204
2.815 0.070 3.063 0.078 0.919 0.0234 306.2569 1204 2.848 0.083
3.103 0.074 0.918 0.0256 581.3344 1202 1.676 0.132 2.082 0.115
0.805 0.0261 817.584 1202 2.181 0.064 2.383 0.060 0.915 0.0261
685.5538 1204 1.862 0.132 2.222 0.083 0.838 0.0263 555.3101 1102
1.640 0.094 1.944 0.093 0.844 0.0277 666.5455 1204 4.533 0.128
4.059 0.168 1.117 0.0301 820.5677 1202 1.550 0.163 2.037 0.145
0.761 0.031 773.5368 1202 1.455 0.109 1.808 0.114 0.805 0.0324
541.3432 1102 3.148 0.073 3.377 0.073 0.932 0.0328 767.547 1202
2.917 0.095 3.237 0.111 0.901 0.0345 569.369 1202 4.824 0.048 4.987
0.057 0.967 0.0352 871.5935 1202 1.702 0.068 2.049 0.145 0.830
0.0362 1226.0968 1203 3.089 0.116 2.709 0.130 1.140 0.0365 793.5679
1204 3.022 0.067 3.237 0.075 0.934 0.0407 411.3211 1202 2.817 0.068
2.988 0.042 0.943 0.0408 684.5491 1204 2.304 0.087 2.600 0.110
0.886 0.041 768.5504 1202 1.786 0.141 2.160 0.106 0.827 0.041
743.5469 1202 2.894 0.076 3.173 0.109 0.912 0.0417 482.3215 1202
1.866 0.054 2.056 0.072 0.908 0.0424 574.4637 1202 1.059 0.201
1.653 0.204 0.640 0.0444 509.3493 1202 2.290 0.057 2.490 0.079
0.919 0.0446 772.586 1204 3.167 0.062 3.330 0.050 0.951 0.0458
383.3283 1204 1.280 0.142 1.662 0.119 0.770 0.0462 664.5322 1204
4.643 0.145 4.176 0.173 1.112 0.0465 312.2312 1204 2.771 0.059
2.583 0.070 1.073 0.0467 481.3172 1202 3.803 0.057 3.982 0.065
0.955 0.0468 765.5335 1204 3.065 0.107 3.418 0.137 0.897 0.0491
847.5953 1202 2.162 0.114 2.518 0.134 0.858 0.0492 624.5131 1203
3.900 0.068 4.084 0.061 0.955 0.0497
TABLE-US-00011 TABLE 11 Accurate mass features differing between
patients showing the best discrimination between high ADAS score
and low ADAS score (p < 0.05, log2 transformed). AVG AVG
Detected Analysis (log2) SEM high (log2) SEM low log(2) Mass Mode
high ADAS ADAS low ADAS ADAS Ratio P Value 541.3435 1102 3.007
0.047 3.741 0.083 0.804 5.76E-09 569.3685 1102 1.862 0.065 2.686
0.070 0.693 9.27E-09 804.5713 1102 3.915 0.047 4.494 0.080 0.871
3.59E-07 803.5681 1102 5.145 0.046 5.720 0.084 0.899 5.31E-07
837.5027 1101 3.257 0.058 3.866 0.070 0.842 5.33E-07 807.5758 1101
5.318 0.076 6.045 0.065 0.880 5.92E-07 808.5792 1101 4.265 0.076
4.992 0.069 0.854 7.70E-07 832.6024 1102 3.129 0.065 3.740 0.063
0.837 1.34E-06 779.5444 1101 6.061 0.061 6.691 0.082 0.906 1.39E-06
780.5474 1101 5.073 0.063 5.714 0.083 0.888 1.42E-06 831.5995 1102
4.245 0.068 4.850 0.061 0.875 2.37E-06 546.3486 1101 1.593 0.073
2.250 0.084 0.708 5.48E-06 871.5529 1102 2.778 0.050 3.307 0.090
0.840 6.59E-06 567.3545 1102 2.637 0.079 3.305 0.081 0.798 8.54E-06
545.3453 1101 3.142 0.085 3.836 0.077 0.819 9.04E-06 805.5604 1101
5.013 0.074 5.685 0.099 0.882 9.51E-06 794.5422 1102 2.287 0.056
2.795 0.079 0.818 1.22E-05 886.5584 1102 2.853 0.053 3.354 0.081
0.851 1.32E-05 827.57 1102 3.893 0.063 4.438 0.079 0.877 1.41E-05
828.5737 1102 2.920 0.061 3.425 0.069 0.852 1.56E-05 805.5831 1102
3.821 0.058 4.385 0.100 0.871 1.61E-05 829.5856 1102 3.781 0.076
4.393 0.077 0.861 1.68E-05 915.5191 1101 2.144 0.053 2.621 0.078
0.818 1.99E-05 793.5387 1102 3.352 0.056 3.854 0.086 0.870 2.76E-05
806.5637 1101 4.001 0.078 4.667 0.107 0.857 2.85E-05 821.5713 1102
2.834 0.067 3.309 0.046 0.856 3.30E-05 565.3396 1102 3.216 0.096
3.992 0.121 0.806 3.57E-05 555.3098 1102 1.608 0.081 2.242 0.094
0.717 4.04E-05 757.4989 1101 3.119 0.086 3.849 0.148 0.810 0.0001
832.5792 1101 3.024 0.092 3.662 0.085 0.826 0.0001 855.6015 1102
3.274 0.068 3.795 0.093 0.862 0.0001 458.2404 1101 1.430 0.076
1.984 0.109 0.721 0.0002 783.5671 1101 3.275 0.105 4.025 0.141
0.813 0.0002 803.5445 1101 4.778 0.095 5.397 0.079 0.885 0.0002
804.5475 1101 3.844 0.095 4.459 0.075 0.862 0.0002 831.576 1101
3.924 0.100 4.575 0.095 0.858 0.0002 518.3174 1101 3.093 0.072
3.609 0.101 0.857 0.0003 781.5616 1101 5.679 0.096 6.315 0.110
0.899 0.0003 517.314 1101 5.122 0.077 5.642 0.104 0.908 0.0004
782.5649 1101 4.649 0.100 5.296 0.118 0.878 0.0004 785.5933 1101
5.333 0.121 6.107 0.132 0.873 0.0004 786.5967 1101 4.256 0.124
5.039 0.133 0.844 0.0004 833.5932 1101 2.746 0.113 3.494 0.134
0.786 0.0004 853.5855 1102 2.333 0.088 2.875 0.080 0.811 0.0004
755.5468 1101 1.879 0.101 2.593 0.157 0.725 0.0005 811.6096 1101
2.512 0.140 3.375 0.152 0.744 0.0006 809.5936 1101 4.448 0.126
5.177 0.119 0.859 0.0007 761.5843 1101 1.993 0.094 2.654 0.160
0.751 0.0008 810.5969 1101 3.451 0.128 4.183 0.115 0.825 0.0008
847.5316 1101 2.543 0.093 3.082 0.092 0.825 0.0008 757.5626 1101
6.144 0.104 6.825 0.155 0.900 0.0009 758.5656 1101 5.185 0.107
5.885 0.158 0.881 0.0009 543.3296 1101 3.486 0.084 4.025 0.125
0.866 0.001 759.5779 1101 5.080 0.100 5.759 0.169 0.882 0.0011
760.5811 1101 3.952 0.102 4.645 0.177 0.851 0.0012 795.5551 1102
2.493 0.075 2.937 0.092 0.849 0.0012 829.5604 1101 3.200 0.098
3.759 0.106 0.851 0.0012 523.3635 1101 2.882 0.117 3.595 0.163
0.801 0.0014 575.2728 1101 2.204 0.078 2.673 0.104 0.824 0.0014
784.581 1101 3.921 0.129 4.667 0.171 0.840 0.0018 819.5553 1102
2.210 0.089 2.665 0.085 0.829 0.0025 817.5378 1102 2.089 0.088
2.491 0.068 0.839 0.0047 731.5463 1101 1.373 0.174 2.218 0.197
0.619 0.0052 795.5183 1101 2.270 0.081 2.725 0.140 0.833 0.0058
722.5334 1101 2.190 0.085 2.598 0.111 0.843 0.0083 760.5217 1204
4.084 0.063 4.331 0.042 0.943 0.0118 833.757 1203 2.531 0.161 3.274
0.231 0.773 0.0132 429.3749 1204 2.703 0.080 3.137 0.167 0.861
0.0143 722.4789 1201 1.895 0.093 2.247 0.064 0.843 0.0145 1228.1111
1203 2.699 0.127 3.291 0.194 0.820 0.0146 541.3139 1101 3.909 0.116
4.412 0.152 0.886 0.0158 860.7753 1203 4.152 0.110 4.647 0.161
0.893 0.016 703.5709 1101 2.858 0.095 3.216 0.072 0.888 0.0163
579.5325 1203 3.776 0.177 4.492 0.191 0.841 0.0166 858.686 1202
3.030 0.064 2.781 0.057 1.090 0.0166 831.7408 1203 3.708 0.137
4.286 0.176 0.865 0.0175 484.3041 1203 2.257 0.120 1.475 0.348
1.530 0.0176 580.5352 1203 1.310 0.222 2.135 0.172 0.614 0.0181
830.7362 1203 4.568 0.137 5.156 0.187 0.886 0.0182 495.332 1101
4.769 0.114 5.305 0.195 0.899 0.0186 183.0662 1101 1.868 0.215
2.671 0.183 0.699 0.0191 702.5677 1101 4.033 0.104 4.409 0.074
0.915 0.0194 887.8001 1203 2.884 0.137 3.396 0.122 0.849 0.0199
886.7916 1203 4.153 0.131 4.631 0.120 0.897 0.0232 759.5164 1204
4.697 0.077 4.975 0.066 0.944 0.0236 725.5527 1101 3.123 0.108
3.499 0.079 0.892 0.0244 832.7522 1203 3.418 0.170 4.119 0.245
0.830 0.0255 828.7207 1203 5.336 0.130 5.867 0.187 0.909 0.0263
496.3355 1101 2.972 0.113 3.473 0.195 0.856 0.0264 604.5432 1203
6.338 0.149 6.863 0.137 0.924 0.0283 864.738 1204 4.641 0.166 5.295
0.232 0.876 0.0298 680.5623 1204 3.590 0.125 4.126 0.214 0.870
0.0305 627.5201 1204 3.756 0.157 4.380 0.227 0.858 0.0315 1019.3838
1102 3.116 0.042 2.950 0.063 1.056 0.0323 825.6927 1203 1.408 0.212
2.138 0.204 0.658 0.0331 863.7336 1204 5.382 0.170 6.035 0.237
0.892 0.0338 521.3477 1101 3.368 0.126 3.890 0.213 0.866 0.0349
731.4916 1204 5.513 0.065 5.746 0.076 0.959 0.0351 542.3173 1101
2.116 0.120 2.565 0.157 0.825 0.0352 829.7242 1204 2.575 0.161
3.185 0.221 0.809 0.0356 835.6998 1204 2.285 0.247 3.153 0.271
0.725 0.0356 276.2095 1204 2.479 0.091 2.885 0.179 0.859 0.0358
653.536 1204 4.096 0.154 4.679 0.218 0.875 0.0368 523.4678 1203
2.747 0.238 3.609 0.295 0.761 0.0372 829.7246 1203 4.606 0.121
5.065 0.173 0.909 0.0376 921.8142 1204 2.618 0.338 3.654 0.184
0.716 0.039 605.5456 1203 4.920 0.154 5.420 0.131 0.908 0.0392
549.4844 1203 5.315 0.129 5.753 0.137 0.924 0.0398 732.4939 1204
4.258 0.065 4.480 0.069 0.950 0.04 365.3159 1203 1.907 0.098 1.449
0.217 1.316 0.0402 867.7581 1204 2.358 0.280 3.299 0.284 0.715
0.0402 738.5445 1102 2.458 0.122 2.825 0.059 0.870 0.0409 859.7695
1203 5.769 0.102 6.185 0.179 0.933 0.0414 625.5075 1204 3.271 0.138
3.811 0.224 0.858 0.0422 512.3356 1203 2.602 0.095 2.151 0.223
1.210 0.0432 842.7387 1203 2.626 0.103 2.998 0.139 0.876 0.0432
552.5021 1203 2.742 0.161 3.315 0.213 0.827 0.0444 724.5493 1101
4.410 0.105 4.736 0.081 0.931 0.0447 626.5108 1204 2.117 0.159
2.677 0.209 0.791 0.0454 928.7505 1203 2.869 0.168 2.238 0.265
1.282 0.0473 894.781 1204 3.884 0.196 4.542 0.228 0.855 0.048
866.7533 1204 3.905 0.219 4.667 0.292 0.837 0.0492 278.2255 1204
4.693 0.070 5.006 0.156 0.937 0.0494 865.7486 1204 4.644 0.223
5.419 0.296 0.857 0.0497
TABLE-US-00012 TABLE 12 Accurate mass features differing between
patients showing the best discrimination between high MMSE score
and low MMSE score (p < 0.05, log2 transformed). AVG AVG (log2)
SEM (log2) SEM Detected Analysis low low high high log(2) Mass Mode
MMSE MMSE MMSE MMSE Ratio P Value 505.3229 1202 3.585 0.045 4.514
0.053 0.794 3.90E-15 857.6186 1202 3.339 0.058 4.298 0.053 0.777
5.05E-14 803.5678 1202 6.747 0.056 7.962 0.082 0.847 5.19E-14
858.6211 1202 2.300 0.058 3.221 0.059 0.714 6.73E-13 566.3434 1202
4.856 0.055 5.682 0.055 0.855 1.90E-12 801.555 1202 2.220 0.057
3.128 0.064 0.710 2.89E-12 832.6036 1202 4.692 0.057 5.569 0.061
0.842 3.64E-12 804.5718 1202 5.516 0.047 6.475 0.080 0.852 4.63E-12
565.3393 1202 6.629 0.054 7.462 0.062 0.888 9.30E-12 506.3214 1202
2.237 0.051 2.908 0.044 0.769 1.26E-11 743.5467 1202 2.635 0.077
3.621 0.066 0.728 2.57E-11 831.6 1202 5.793 0.055 6.774 0.087 0.855
4.69E-11 744.55 1202 1.608 0.082 2.504 0.056 0.642 1.72E-10
570.3724 1202 2.702 0.042 3.289 0.052 0.821 4.09E-10 829.5859 1202
5.572 0.065 6.476 0.085 0.860 8.42E-10 830.5894 1202 4.477 0.061
5.311 0.078 0.843 1.01E-09 509.3494 1202 2.113 0.042 2.685 0.059
0.787 4.17E-09 542.3461 1202 3.562 0.047 4.292 0.082 0.830 6.29E-09
711.2577 1202 1.891 0.072 2.585 0.057 0.731 1.12E-08 709.2595 1202
1.864 0.079 2.546 0.047 0.732 1.49E-08 481.3172 1202 3.642 0.055
4.233 0.060 0.860 2.74E-08 569.369 1202 4.664 0.046 5.203 0.063
0.897 7.82E-08 805.5839 1202 5.216 0.051 5.867 0.082 0.889 1.16E-07
806.5873 1202 4.024 0.052 4.658 0.078 0.864 1.16E-07 856.6061 1202
4.179 0.069 5.099 0.121 0.819 1.81E-07 743.5469 1204 5.166 0.051
5.635 0.049 0.917 1.96E-07 793.5386 1102 2.966 0.065 3.468 0.043
0.855 2.63E-07 855.6023 1202 5.166 0.065 6.090 0.128 0.848 2.81E-07
482.3215 1202 1.742 0.057 2.291 0.066 0.761 4.15E-07 541.3433 1202
5.318 0.067 6.197 0.123 0.858 4.56E-07 744.5535 1204 4.203 0.048
4.631 0.048 0.908 4.68E-07 807.5899 1202 2.124 0.071 2.730 0.070
0.778 8.81E-07 827.5701 1202 5.937 0.077 6.931 0.147 0.857 1.20E-06
847.5954 1202 2.030 0.095 2.867 0.104 0.708 1.37E-06 828.5742 1202
4.908 0.074 5.818 0.137 0.844 1.67E-06 787.5464 1204 2.601 0.059
3.069 0.056 0.847 2.17E-06 591.3542 1202 3.749 0.081 4.387 0.078
0.855 3.05E-06 794.5422 1102 1.910 0.065 2.338 0.040 0.817 3.76E-06
804.5713 1102 3.619 0.065 4.092 0.056 0.885 4.94E-06 803.568 1102
4.829 0.068 5.317 0.057 0.908 5.22E-06 758.5094 1204 4.437 0.052
4.813 0.046 0.922 5.89E-06 592.3571 1202 2.013 0.084 2.634 0.079
0.764 7.14E-06 760.5216 1204 3.930 0.056 4.313 0.045 0.911 8.02E-06
759.5165 1204 4.558 0.067 5.001 0.050 0.911 9.01E-06 767.5469 1202
2.700 0.119 3.526 0.101 0.766 9.49E-06 786.5416 1204 3.664 0.051
4.093 0.064 0.895 1.12E-05 784.5239 1204 3.102 0.069 3.575 0.060
0.868 1.23E-05 821.5711 1102 2.415 0.067 2.863 0.055 0.843 1.25E-05
819.5641 1202 2.451 0.109 3.287 0.126 0.746 2.03E-05 777.5531 1202
1.473 0.133 2.257 0.083 0.653 2.21E-05 853.5862 1202 4.524 0.069
5.222 0.126 0.866 3.29E-05 507.3316 1202 2.653 0.056 3.061 0.064
0.867 3.70E-05 731.4917 1204 5.356 0.062 5.744 0.053 0.932 4.18E-05
831.5998 1102 3.958 0.081 4.429 0.057 0.894 4.26E-05 854.5902 1202
3.521 0.075 4.231 0.130 0.832 4.70E-05 630.5586 1203 2.845 0.054
3.242 0.069 0.877 0.0001 732.494 1204 4.127 0.062 4.492 0.050 0.919
0.0001 741.5307 1204 2.661 0.099 3.337 0.105 0.797 0.0001 771.5815
1204 4.036 0.064 4.423 0.055 0.913 0.0001 819.5552 1102 1.733 0.081
2.149 0.045 0.806 0.0001 832.6027 1102 2.864 0.081 3.318 0.056
0.863 0.0001 871.5527 1102 2.467 0.073 2.914 0.066 0.846 0.0001
871.5935 1202 1.550 0.108 2.224 0.110 0.697 0.0001 886.7917 1203
3.826 0.095 4.459 0.099 0.858 0.0001 910.7968 1203 3.257 0.083
3.757 0.079 0.867 0.0001 529.3166 1202 2.771 0.071 3.332 0.112
0.832 0.0002 589.3404 1202 5.578 0.072 6.318 0.155 0.883 0.0002
768.5503 1202 1.668 0.144 2.412 0.103 0.691 0.0002 884.7801 1203
6.076 0.082 6.639 0.104 0.915 0.0002 568.3574 1202 3.760 0.055
4.144 0.074 0.907 0.0003 685.2601 1202 1.483 0.102 2.052 0.093
0.723 0.0003 765.5313 1202 1.447 0.148 2.219 0.120 0.652 0.0003
772.5863 1204 3.177 0.051 3.466 0.050 0.917 0.0003 829.5856 1102
3.407 0.088 3.862 0.064 0.882 0.0003 911.8032 1203 2.388 0.077
2.794 0.064 0.855 0.0003 786.5967 1101 4.302 0.096 4.955 0.132
0.868 0.0004 885.7854 1203 5.116 0.080 5.615 0.095 0.911 0.0004
174.1409 1203 1.904 0.122 2.872 0.212 0.663 0.0005 590.3431 1202
3.790 0.071 4.415 0.142 0.858 0.0005 604.5432 1203 6.172 0.114
6.803 0.113 0.907 0.0005 757.5626 1101 6.170 0.099 6.804 0.129
0.907 0.0005 879.5999 1202 2.601 0.104 3.235 0.124 0.804 0.0005
605.5456 1203 4.762 0.115 5.393 0.118 0.883 0.0006 757.5016 1204
3.549 0.094 3.977 0.057 0.893 0.0006 581.3345 1202 1.572 0.124
2.253 0.131 0.698 0.0007 628.5421 1203 3.231 0.070 3.606 0.070
0.896 0.0007 723.5195 1202 2.196 0.085 2.730 0.110 0.805 0.0007
749.5365 1202 3.146 0.084 3.579 0.078 0.879 0.0007 785.5933 1101
5.382 0.098 6.025 0.137 0.893 0.0007 887.7999 1203 2.493 0.171
3.227 0.087 0.773 0.0007 886.8012 1204 3.209 0.117 3.782 0.096
0.849 0.0008 186.1408 1203 1.539 0.100 2.279 0.172 0.675 0.0009
200.1564 1203 2.724 0.075 3.280 0.129 0.830 0.0009 769.5655 1204
3.833 0.075 4.197 0.063 0.913 0.0009 817.5838 1202 2.041 0.086
2.452 0.070 0.833 0.0009 715.5169 1204 2.238 0.200 3.073 0.108
0.728 0.001 699.52 1204 2.223 0.082 2.690 0.099 0.826 0.0011
758.5656 1101 5.231 0.101 5.844 0.134 0.895 0.0011 795.5182 1101
2.324 0.087 2.800 0.098 0.830 0.0011 772.528 1204 3.202 0.075 3.531
0.053 0.907 0.0012 739.5164 1202 1.109 0.170 1.859 0.124 0.597
0.0014 779.5444 1101 6.186 0.098 6.639 0.083 0.932 0.0014 244.2187
1203 2.645 0.062 3.037 0.093 0.871 0.0015 508.3356 1202 0.969 0.088
1.386 0.078 0.700 0.0015 811.6096 1101 2.569 0.101 3.204 0.152
0.802 0.0017 882.766 1203 7.238 0.092 7.755 0.117 0.933 0.0017
744.4954 1204 3.832 0.067 4.118 0.049 0.930 0.0018 884.7877 1204
4.658 0.101 5.127 0.092 0.909 0.0018 262.2293 1203 2.639 0.059
2.992 0.084 0.882 0.002 784.5811 1101 3.896 0.100 4.493 0.143 0.867
0.002 817.5375 1102 1.715 0.076 2.148 0.101 0.798 0.002 723.5197
1204 3.643 0.087 4.211 0.143 0.865 0.0021 810.5401 1204 2.693 0.070
3.119 0.104 0.863 0.0021 880.6035 1202 1.837 0.084 2.351 0.126
0.782 0.0021 807.5757 1101 5.434 0.097 5.912 0.102 0.919 0.0022
883.7705 1203 6.396 0.085 6.870 0.111 0.931 0.0022 749.576 1102
2.516 0.057 2.174 0.084 1.157 0.0023 852.5738 1202 3.365 0.120
3.968 0.135 0.848 0.0025 752.5564 1202 2.277 0.101 2.730 0.092
0.834 0.0027 783.5672 1101 3.273 0.106 3.856 0.141 0.849 0.0027
808.5792 1101 4.393 0.099 4.868 0.104 0.902 0.0027 755.5468 1101
1.885 0.107 2.412 0.121 0.782 0.0031 780.5475 1101 5.202 0.096
5.629 0.089 0.924 0.0031 826.707 1204 2.266 0.118 2.743 0.086 0.826
0.0031 851.5698 1202 4.297 0.118 4.898 0.142 0.877 0.0031 631.5619
1203 1.475 0.132 1.956 0.066 0.754 0.0033 755.4861 1204 3.019 0.074
3.424 0.102 0.882 0.0034 188.1566 1203 2.272 0.134 2.926 0.154
0.777 0.0035 883.7765 1204 4.148 0.099 4.589 0.095 0.904 0.0035
726.5459 1204 2.790 0.064 3.090 0.069 0.903 0.0038 750.5403 1202
2.185 0.089 2.549 0.073 0.857 0.0038 627.5302 1203 2.497 0.080
2.845 0.075 0.878 0.0039 885.7922 1204 3.954 0.103 4.382 0.089
0.902 0.0039 675.6357 1203 1.815 0.210 0.815 0.240 2.228 0.0042
751.5529 1202 3.374 0.098 3.787 0.090 0.891 0.0043 882.7723 1204
4.984 0.102 5.422 0.096 0.919 0.0043 530.3213 1202 1.209 0.089
1.698 0.129 0.712 0.0044 380.3096 1204 1.432 0.107 1.867 0.092
0.767 0.0046 783.5148 1204 3.121 0.064 3.517 0.111 0.887 0.0048
202.172 1203 3.048 0.112 3.631 0.153 0.839 0.0049 748.5721 1102
3.632 0.058 3.325 0.082 1.092 0.0049 603.5317 1203 7.585 0.077
7.883 0.060 0.962 0.005 243.0718 1101 4.430 0.144 3.210 0.373 1.380
0.0052 582.2473 1201 3.377 0.132 2.673 0.191 1.264 0.0055 739.5141
1204 2.498 0.195 3.173 0.109 0.787 0.0056 879.7454 1204 3.291 0.124
3.749 0.088 0.878 0.0057 494.4342 1203 1.382 0.353 2.777 0.303
0.497 0.0059 782.5086 1204 3.266 0.073 3.662 0.110 0.892 0.0059
855.6016 1102 2.984 0.080 3.363 0.097 0.887 0.006 921.8153 1204
2.175 0.312 3.187 0.131 0.682 0.006 724.5256 1204 2.877 0.077 3.324
0.128 0.866 0.0061 183.0661 1101 2.155 0.151 2.748 0.130 0.784
0.0065 522.4634 1203 4.325 0.238 5.237 0.198 0.826 0.0067 853.7296
1204 3.607 0.109 4.012 0.083 0.899 0.0068 216.1877 1203 2.796 0.092
3.282 0.140 0.852 0.0076 779.5442 1201 7.005 0.079 7.301 0.065
0.959 0.0076 777.5288 1201 2.657 0.093 3.009 0.079 0.883 0.0077
523.468 1203 2.895 0.227 3.731 0.183 0.776 0.0083 569.3687 1102
1.734 0.098 2.075 0.067 0.836 0.0085 775.5535 1202 2.112 0.083
2.438 0.079 0.866 0.0087 632.5035 1204 4.858 0.122 5.325 0.110
0.912 0.0088 805.5604 1101 5.112 0.099 5.504 0.097 0.929 0.0089
313.269 1203 1.592 0.175 2.180 0.112 0.730 0.009 743.5464 1203
1.703 0.150 2.156 0.056 0.790 0.009 768.5539 1204 3.881 0.077 4.240
0.100 0.915 0.009 686.488 1204 2.671 0.054 2.872 0.047 0.930 0.0092
886.5582 1102 2.503 0.058 2.781 0.079 0.900 0.0092 602.5282 1203
8.833 0.081 9.116 0.059 0.969 0.0093 558.4667 1202 1.932 0.201
2.652 0.159 0.728 0.0095 809.5936 1101 4.523 0.112 5.054 0.155
0.895 0.0101 881.7616 1204 4.015 0.111 4.427 0.099 0.907 0.0101
833.7558 1204 1.919 0.311 2.863 0.138 0.670 0.0103 864.7596 1203
2.184 0.067 2.468 0.078 0.885 0.0103 613.3405 1202 3.422 0.124
3.963 0.150 0.863 0.0104 626.5277 1203 3.649 0.073 3.946 0.079
0.925 0.0105 629.5449 1203 2.015 0.088 2.370 0.093 0.850 0.0106
767.5494 1204 4.975 0.083 5.343 0.104 0.931 0.0106 517.3722 1204
2.176 0.149 1.408 0.235 1.545 0.0107 773.5347 1204 3.151 0.056
3.426 0.082 0.920 0.0108 806.5638 1101 4.113 0.100 4.498 0.097
0.914 0.0109 204.1876 1203 2.205 0.099 2.682 0.143 0.822 0.0113
837.5027 1101 3.439 0.075 3.734 0.078 0.921 0.0116 595.4932 1202
1.393 0.217 2.170 0.185 0.642 0.0117 915.5192 1101 2.301 0.087
2.588 0.060 0.889 0.0117 257.8105 1101 3.636 0.084 2.677 0.344
1.358 0.0121 541.343 1102 2.816 0.087 3.105 0.062 0.907 0.0121
745.5656 1204 3.838 0.051 4.054 0.062 0.947 0.0125 749.5408 1204
4.465 0.071 4.865 0.130 0.918 0.0126 725.5385 1204 2.932 0.068
3.293 0.115 0.890 0.0128 757.4993 1101 3.262 0.095 3.716 0.140
0.878 0.0128 852.725 1204 3.774 0.105 4.122 0.076 0.916 0.0128
780.5474 1201 5.941 0.081 6.223 0.067 0.955 0.0129 880.7516 1203
7.205 0.107 7.651 0.127 0.942 0.0129 881.7558 1203 6.471 0.103
6.883 0.116 0.940 0.0132 338.2821 1203 4.474 0.095 4.792 0.073
0.934 0.0135 827.5699 1102 3.597 0.089 3.987 0.116 0.902 0.0136
880.7566 1204 4.483 0.117 4.882 0.094 0.918 0.0138 572.4468 1204
1.726 0.131 2.213 0.129 0.780 0.014 336.2662 1203 3.181 0.097 3.518
0.083 0.904 0.0141 615.3539 1202 2.263 0.089 2.598 0.092 0.871
0.0146 544.4479 1203 1.928 0.085 2.273 0.100 0.848 0.0148 579.5325
1203 3.737 0.170 4.362 0.166 0.857 0.0148 520.45 1203 3.613 0.209
4.378 0.204 0.825 0.0149 633.5078 1204 3.891 0.128 4.347 0.118
0.895 0.0149 519.3321 1101 3.416 0.099 3.875 0.146 0.881 0.0151
547.6031 1101 2.454 0.114 1.714 0.259 1.432 0.0152 727.5569 1204
3.548 0.077 3.866 0.093 0.918 0.0152 339.285 1203 2.242 0.107 2.601
0.087 0.862 0.0154 746.5719 1204 3.004 0.054 3.196 0.050 0.940
0.0154 828.5736 1102 2.605 0.083 2.966 0.111 0.878 0.0154 798.6742
1203 1.685 0.154 2.329 0.192 0.723 0.0155 860.7753 1203 4.054 0.129
4.507 0.117 0.899 0.0156 521.4522 1203 1.837 0.271 2.786 0.244
0.660 0.0158 594.4878 1202 2.699 0.253 3.523 0.191 0.766 0.0158
781.5617 1101 5.737 0.110 6.187 0.134 0.927 0.0158 810.5969 1101
3.498 0.121 4.023 0.161 0.870 0.0158 631.4902 1204 2.690 0.150
3.208 0.132 0.839 0.0163 518.4346 1203 1.198 0.238 2.121 0.267
0.565 0.0164 244.0559 1101 3.875 0.099 3.359 0.174 1.153 0.0165
751.5556 1204 4.690 0.092 5.091 0.126 0.921 0.0165 825.5544 1202
2.647 0.146 3.202 0.159 0.827 0.017 520.3353 1101 1.661 0.103 2.114
0.144 0.786 0.0176 858.7632 1203 6.769 0.125 7.244 0.137 0.935
0.0178 861.7801 1203 2.123 0.171 2.681 0.138 0.792 0.0179 646.4837
1204 1.688 0.192 2.245 0.108 0.752 0.0184 489.6451 1101 2.555 0.109
1.822 0.269 1.402 0.0186 255.8135 1101 4.518 0.090 3.502 0.393
1.290 0.019 260.2135 1203 1.422 0.129 1.878 0.128 0.757 0.0193
833.5931 1101 2.952 0.114 3.347 0.109 0.882 0.0197 831.5759 1101
4.038 0.114 4.451 0.121 0.907 0.0201 544.3971 1204 3.313 0.158
2.377 0.342 1.394 0.0203 294.2207 1204 5.248 0.117 5.652 0.112
0.929 0.0205 724.5493 1101 4.674 0.050 4.395 0.101 1.063 0.0205
663.487 1204 1.733 0.223 2.369 0.128 0.731 0.0207 860.7753 1204
3.460 0.170 3.955 0.104 0.875 0.0207 246.1465 1202 3.707 0.141
4.188 0.134 0.885 0.0209 606.4869 1204 4.562 0.137 5.023 0.126
0.908 0.0215 576.5097 1203 9.304 0.133 9.756 0.127 0.954 0.0216
311.775 1101 2.594 0.152 1.849 0.264 1.403 0.022
566.413 1204 2.026 0.190 2.553 0.100 0.794 0.0221 577.5136 1203
7.959 0.125 8.392 0.124 0.948 0.0224 835.7001 1204 2.246 0.249
3.007 0.187 0.747 0.0226 665.501 1204 2.974 0.116 3.370 0.114 0.883
0.0227 675.6377 1204 3.825 0.077 4.086 0.075 0.936 0.023 600.5127
1203 8.597 0.082 8.866 0.075 0.970 0.0232 752.5583 1204 3.555 0.096
3.939 0.126 0.902 0.0241 161.1053 1101 4.011 0.148 2.977 0.402
1.347 0.0242 616.4673 1201 1.329 0.218 2.031 0.193 0.654 0.0245
253.8163 1101 4.184 0.090 3.283 0.364 1.274 0.0248 878.74 1204
3.665 0.118 4.001 0.074 0.916 0.025 580.535 1203 1.221 0.219 1.936
0.203 0.631 0.0252 859.7694 1203 5.668 0.122 6.076 0.119 0.933
0.0252 538.4224 1202 1.087 0.172 1.689 0.184 0.643 0.0256 804.723
1203 1.425 0.264 2.238 0.214 0.637 0.0256 833.7571 1203 2.599 0.192
3.201 0.165 0.812 0.0261 530.382 1204 2.988 0.204 2.114 0.305 1.413
0.0262 908.7832 1203 5.011 0.090 5.309 0.088 0.944 0.027 653.5361
1204 3.973 0.129 4.367 0.107 0.910 0.0273 559.47 1202 0.812 0.174
1.388 0.171 0.585 0.0274 748.5735 1202 3.935 0.071 3.726 0.054
1.056 0.0279 803.5445 1101 4.899 0.121 5.305 0.124 0.924 0.028
832.7521 1203 3.434 0.205 4.059 0.169 0.846 0.0283 320.2357 1204
1.225 0.148 1.609 0.071 0.761 0.0284 728.5626 1204 2.941 0.066
3.161 0.067 0.930 0.0284 795.555 1102 2.053 0.080 2.292 0.062 0.896
0.0285 838.7232 1204 2.011 0.253 2.753 0.191 0.730 0.0285 782.565
1101 4.709 0.114 5.143 0.147 0.916 0.0286 750.544 1204 3.351 0.066
3.678 0.124 0.911 0.0287 783.4315 1101 2.505 0.125 1.807 0.271
1.386 0.029 214.172 1203 3.549 0.090 3.910 0.126 0.908 0.0292
312.2663 1203 3.879 0.140 4.315 0.126 0.899 0.03 857.7531 1203
7.332 0.109 7.738 0.138 0.947 0.03 202.0453 1101 5.911 0.122 5.238
0.266 1.128 0.0305 763.5157 1202 1.570 0.175 2.098 0.147 0.748
0.0306 218.0192 1101 4.023 0.134 3.143 0.357 1.280 0.0307 856.7481
1203 8.173 0.118 8.608 0.151 0.949 0.032 795.5839 1204 3.545 0.061
3.800 0.093 0.933 0.0321 565.4104 1204 3.353 0.123 3.754 0.127
0.893 0.0332 313.7722 1101 3.080 0.178 2.250 0.319 1.369 0.0334
909.7882 1203 4.177 0.092 4.456 0.081 0.938 0.0337 429.6888 1101
2.475 0.093 1.833 0.268 1.351 0.0341 431.957 1202 1.847 0.154 2.312
0.136 0.799 0.0341 607.4919 1204 2.906 0.259 3.579 0.147 0.812
0.0341 294.1443 1201 2.449 0.210 1.795 0.201 1.364 0.0348 598.4965
1203 6.590 0.104 6.923 0.106 0.952 0.035 549.6005 1101 2.027 0.167
1.424 0.212 1.424 0.0351 283.9028 1101 3.018 0.076 2.397 0.267
1.259 0.0353 820.5679 1202 1.438 0.153 2.061 0.231 0.698 0.0353
574.4597 1204 3.354 0.210 3.994 0.196 0.840 0.0359 826.7048 1203
4.509 0.145 4.950 0.134 0.911 0.0361 805.5832 1102 3.502 0.085
3.742 0.066 0.936 0.0364 550.4956 1203 6.603 0.188 7.165 0.167
0.922 0.0365 630.4861 1204 3.807 0.132 4.238 0.142 0.898 0.0369
551.4985 1203 5.182 0.182 5.733 0.167 0.904 0.0371 549.4845 1203
5.363 0.140 5.765 0.115 0.930 0.0372 827.7083 1203 3.751 0.145
4.174 0.124 0.899 0.0374 306.2569 1204 2.821 0.075 3.051 0.072
0.925 0.0375 775.553 1204 3.272 0.071 3.577 0.119 0.915 0.0377
242.203 1203 4.260 0.087 4.573 0.113 0.932 0.0382 828.7202 1204
3.465 0.158 3.906 0.122 0.887 0.0385 232.2188 1203 3.156 0.098
3.499 0.122 0.902 0.0387 722.4789 1201 1.873 0.084 2.132 0.082
0.879 0.0389 530.3471 1202 2.566 0.117 2.910 0.103 0.882 0.0393
264.2452 1203 1.753 0.138 2.114 0.090 0.829 0.0397 858.7663 1204
5.791 0.140 6.169 0.100 0.939 0.0401 759.5779 1101 5.093 0.108
5.471 0.136 0.931 0.0404 559.469 1204 2.761 0.191 3.288 0.149 0.840
0.0406 593.4743 1204 2.069 0.240 2.728 0.184 0.758 0.0406 392.2938
1204 1.941 0.277 2.718 0.225 0.714 0.041 575.499 1203 7.133 0.096
7.452 0.112 0.957 0.0414 826.5581 1202 1.759 0.125 2.203 0.163
0.798 0.0417 719.6233 1204 4.578 0.094 4.279 0.102 1.070 0.0419
295.2287 1204 2.883 0.203 3.414 0.138 0.844 0.0422 555.3101 1102
1.310 0.070 1.541 0.080 0.850 0.0424 830.7355 1204 3.947 0.183
4.447 0.141 0.888 0.0427 474.3706 1203 0.868 0.128 1.234 0.112
0.704 0.0433 203.1157 1101 3.694 0.116 2.965 0.319 1.246 0.0437
295.2239 1204 3.109 0.123 3.476 0.119 0.894 0.0437 855.7417 1204
4.834 0.122 5.184 0.107 0.932 0.0438 760.5811 1101 3.973 0.109
4.349 0.138 0.913 0.0441 625.5076 1204 3.218 0.140 3.624 0.126
0.888 0.0443 270.2343 1203 1.714 0.106 2.018 0.095 0.850 0.0444
661.6233 1204 2.375 0.073 2.644 0.103 0.898 0.0445 446.3403 1202
0.751 0.166 1.275 0.181 0.589 0.0446 521.3474 1201 2.493 0.169
2.031 0.137 1.228 0.0451 837.718 1204 2.369 0.329 3.272 0.267 0.724
0.0451 228.1877 1203 2.954 0.108 3.313 0.131 0.892 0.0458 832.7495
1204 2.994 0.226 3.588 0.164 0.834 0.0458 576.477 1202 1.734 0.213
2.337 0.187 0.742 0.0459 1085.3294 1101 3.756 0.022 3.463 0.136
1.085 0.0467 444.2717 1202 1.195 0.096 1.496 0.105 0.799 0.0473
545.6062 1101 1.955 0.134 1.427 0.213 1.370 0.0477 651.5221 1204
3.677 0.131 4.039 0.111 0.910 0.0482 829.7244 1203 4.651 0.135
5.044 0.130 0.922 0.0482 859.7706 1204 4.962 0.145 5.331 0.100
0.931 0.0482 575.4632 1204 2.109 0.207 2.692 0.185 0.783 0.0485
804.5476 1101 3.984 0.119 4.353 0.130 0.915 0.0485 725.5527 1101
3.407 0.058 3.160 0.102 1.078 0.0486 350.2423 1202 2.544 0.227
1.901 0.207 1.338 0.0489 382.1083 1101 2.600 0.210 1.858 0.286
1.399 0.049 836.7076 1204 1.427 0.242 2.163 0.256 0.660 0.0493
671.5726 1204 2.055 0.138 2.458 0.136 0.836 0.0495 197.8549 1101
3.431 0.102 2.697 0.337 1.272 0.0499 773.537 1202 1.569 0.078 1.805
0.082 0.869 0.0499
TABLE-US-00013 TABLE 13 Accurate mass features differing between
clinically diagnosed AD and non-AD patients in CSF (p < 0.05,
log2 transformed). AVG AVG Detected Analysis (log2) SEM (log2) SEM
log(2) Mass Mode AD AD non-AD non-AD Ratio P Value 742.2972 1203
1.187 0.130 0.000 0.000 1.187 1.69E-05 562.46 1203 1.080 0.134
0.000 0.000 1.080 4.84E-05 731.653 1203 0.905 0.193 0.000 0.000
0.905 0.0022 432.1532 1203 1.445 0.125 0.880 0.053 0.609 0.0037
487.6482 1101 2.388 0.044 2.668 0.057 1.117 0.0037 275.8712 1101
2.362 0.057 2.664 0.058 1.128 0.0045 371.7311 1101 3.417 0.098
3.818 0.022 1.117 0.0053 622.2539 1203 2.667 0.110 2.166 0.074
0.812 0.0056 485.6503 1101 1.758 0.051 1.976 0.024 1.124 0.0058
207.0822 1203 1.010 0.126 0.198 0.198 0.196 0.006 640.2637 1203
3.557 0.152 2.876 0.106 0.809 0.0063 373.728 1101 3.013 0.086 3.358
0.037 1.114 0.0071 656.2587 1203 0.977 0.080 0.322 0.199 0.330
0.0097 730.6493 1203 1.888 0.139 0.948 0.278 0.502 0.0109 220.0798
1101 2.252 0.078 2.556 0.044 1.135 0.011 641.2661 1203 1.915 0.148
1.010 0.282 0.527 0.0152 779.4393 1101 3.612 0.032 3.728 0.022
1.032 0.0167 782.5647 1201 1.898 0.208 1.128 0.142 0.594 0.0172
313.7721 1101 4.045 0.096 4.350 0.031 1.075 0.0215 785.4288 1101
3.273 0.039 3.410 0.033 1.042 0.0253 777.4426 1101 2.495 0.017
2.614 0.044 1.048 0.0279 341.8614 1101 2.747 0.069 2.990 0.061
1.089 0.0305 250.0366 1101 3.122 0.115 3.462 0.049 1.109 0.0309
315.7693 1101 3.347 0.090 3.618 0.044 1.081 0.0311 253.8164 1101
4.237 0.108 4.542 0.030 1.072 0.033 369.7338 1101 2.713 0.148 3.132
0.035 1.154 0.034 781.5607 1201 2.728 0.234 2.048 0.095 0.751
0.0345 206.0789 1203 4.178 0.147 3.484 0.253 0.834 0.0357 429.689
1101 3.232 0.042 3.392 0.051 1.050 0.0358 638.2465 1203 1.927 0.188
1.372 0.095 0.712 0.0362 578.1574 1101 1.828 0.042 1.674 0.046
0.916 0.0368 262.0777 1101 4.738 0.114 5.072 0.052 1.070 0.0371
265.8423 1101 3.682 0.101 3.960 0.028 1.076 0.0395 582.6334 1101
1.702 0.051 1.526 0.052 0.897 0.0395 255.8135 1101 4.678 0.113
4.978 0.029 1.064 0.0418 321.8039 1101 3.190 0.098 3.480 0.062
1.091 0.0423 514.6879 1101 2.650 0.037 2.534 0.032 0.956 0.0424
262.0221 1101 3.633 0.126 3.964 0.026 1.091 0.0435 746.621 1101
1.015 0.225 0.306 0.190 0.301 0.0438 781.4354 1101 4.163 0.028
4.254 0.025 1.022 0.045 311.775 1101 3.457 0.111 3.764 0.053 1.089
0.0461 257.8105 1101 3.753 0.115 4.056 0.036 1.081 0.0472
TABLE-US-00014 TABLE 14 Retention Time of Six Biomarkers Q-Star
Retention FT Accurate Theoretical Mass Time Neutral Mass Formula
Neutral Mass (M - H) ion (min) 541.3432 C25H52NO9P 541.3379
540.3616 29.4 569.3687 C27H56NO9P 569.3692 568.3979 31.0 699.5198
C39H74NO7P 699.5202 698.5392 40.5 723.5195 C41H74NO7P 723.5202
722.5331 40.2 751.5555 C43H78NO7P 751.5515 750.5667 41.9 803.568
C43H82NO10P 803.5676 802.5575 38.6
TABLE-US-00015 TABLE 15 Metabolite 541.3432 Fragments Frag Q-Star
Formula Theoretical Detected Delta Diff Loss C25H51NO9P 540.33014
540.3616 0.0315 -- -- C23H47NO7P 480.30901 480.3313 0.0223 60.02112
C2H4O2 C16H31O2 255.23240 255.2521 0.0197 292.15251 C9H27NO7P
C7H15NO5P 224.06878 224.0904 0.0216 300.26644 C18H36O3 O3P 78.95850
78.975 0.0165 461.37163 C25H51NO6 Legend: frag formula: The
putative computationally derived molecular formula of the fragment
neutral mass. theoretical: The theoretical mass of the formulas
shown in the frag formula column. Qstar-detected: The detected mass
from the ABI Q-Star XL. delta: The difference between the
theoretical and neutral mass. diff: The mass difference between the
Qstar-detected parent ion mass and the Qstar-detected fragmant ion
mass. Loss: The putative molecular formulas of the "diff" column
Note: These are only predicted formulas for each fragment and are
not necessarily the actual formulas.
TABLE-US-00016 TABLE 16 Metabolite 569.3687 Fragments Q-Star Frag
Formula Theoretical Detected Delta Diff Loss C27H55NO9P 568.3614
568.3979 0.0365 -- -- C25H51NO7P 508.34031 508.3612 0.0209 60.02112
C2H4O2 C18H35O2 283.26370 283.2900 0.0263 285.09773 C9H20NO7P
C7H15NO5P 224.06878 224.0899 0.0211 344.23266 C20H40O4 C4H11NO4P
168.04256 168.0668 0.0242 400.31887 C23H44O5 C3H6PO5 152.99528
153.0196 0.0243 446.33992 C24H49NO4P O3P 78.95850 78.975 0.0165
489.40293 C27H55NO6 Legend: frag formula: The putative
computationally derived molecular formula of the fragment neutral
mass. theoretical: The theoretical mass of the formulas shown in
the frag formula column. Qstar-detected: The detected mass from the
ABI Q-Star XL. delta: The difference between the theoretical and
neutral mass. diff: The mass difference between the Qstar-detected
parent ion mass and the Qstar-detected fragmant ion mass. Loss: The
putative molecular formulas of the "diff" column Note: These are
only predicted formulas for each fragment and are not necessarily
the actual formulas.
TABLE-US-00017 TABLE 17 Metabolite 803.568 Fragments Q-Star Frag
Formula Theoretical Detected Delta Differences Loss C43H81NO10P
802.55980 802.5575 -0.0023 -- -- C41H77NO8P 742.53868 742.5526
0.0139 76.01604 C2H4O3 C23H47NO7P 480.30901 480.3248 0.0158
322.25079 C20H34O3 C18H32O2 279.23240 279.2485 0.0161 522.31957
C25H49NO8P C16H31O2 255.23240 255.2469 0.0145 547.32740 C27H50NO8P
C7H15NO5P 224.06878 224.0853 0.0165 578.49102 C36H66O5 C4H11NO4P
168.04256 168.0629 0.0203 634.51724 C39H70O6 Legend: frag formula:
The putative computationally derived molecular formula of the
fragment neutral mass. theoretical: The theoretical mass of the
formulas shown in the frag formula column. Qstar-detected: The
detected mass from the ABI Q-Star XL. delta: The difference between
the theoretical and neutral mass. diff: The mass difference between
the Qstar-detected parent ion mass and the Qstar-detected fragmant
ion mass. Loss: The putative molecular formulas of the "diff"
column Note: These are only predicted formulas for each fragment
and are not necessarily the actual formula
TABLE-US-00018 TABLE 18 List of Preferred List of Metabolites
Metabolite Code Metabolite Name Molecular Formula Parent Mass M - H
Mass Diagnostic Fragment Mass MS/MS Transition M01 PtdEt 16:0/18:0
C39H78N1O8P1 719.54648 718.5 R1 (C16H31O2) - 255 718.0/255.0 M02
PtdEt 16:0/18:1 C39H76N1O8P1 717.53083 716.5 R1 (C16H31O2) - 255
716.0/255.0 M03 PtdEt 18:0/18:0 C41H82N1O8P1 747.57777 746.5 R1
(C18H35O2) - 283 746.0/283.0 M04 PtdEt 18:0/18:1 C41H80N1O8P1
745.56213 744.5 R1 (C18H35O2) - 283 744.0/283.0 M05 Plasmanyl
16:0/18:1 C39H78N1O7P1 703.55156 702.5 R2 (C18H33O2) - 281
702.0/281.0 M06 Plasmanyl 16:0/18:2 C39H76N1O7P1 701.53591 700.5 R2
(C18H31O2) - 279 700.0/279.0 M07 Plasmanyl 16:0/20:4 C41H76N1O7P1
725.53591 724.5 R2 (C20H31O2) - 303 724.0/303.0 M08 Plasmanyl
16:0/22:4 C43H80N1O7P1 753.56721 752.5 R2 (C22H35O2) - 331
752.0/331.0 M09 Plasmanyl 16:0/22:6 C43H76N1O7P1 749.53591 748.5 R2
(C22H31O2) - 327 748.0/327.0 M10 Plasmanyl 18:0/18:1 C41H82N1O7P1
731.58286 730.5 R2 (C18H33O2) - 281 730.0/281.0 M11 Plasmanyl
18:0/18:2 C41H80N1O7P1 729.56721 728.5 R2 (C18H31O2) - 279
728.0/279.0 M12 Plasmanyl 18:0/20:4 C43H80N1O7P1 753.56721 752.5 R2
(C20H31O2) - 303 752.0/303.0 M13 Plasmanyl 18:0/22:4 C45H84N1O7P1
781.59851 780.5 R2 (C22H35O2) - 331 780.0/331.0 M14 Plasmanyl
18:0/22:6 C45H80N1O7P1 777.56721 776.5 R2 (C22H31O2) - 327
776.0/327.0 M15 Plasmenyl 16:0/18:1 C39H76N1O7P1 701.53591 700.5 R2
(C18H33O2) - 281 700.0/281.0 M16 Plasmenyl 16:0/18:2 C39H74N1O7P1
699.52026 698.5 R2 (C18H31O2) - 279 698.0/279.0 M17 Plasmenyl
16:0/20:4 C41H74N1O7P1 723.52026 722.5 R2 (C20H31O2) - 303
722.0/303.0 M18 Plasmenyl 16:0/22:4 C43H78N1O7P1 751.55156 750.5 R2
(C22H35O2) - 331 750.0/331.0 M19 Plasmenyl 16:0/22:6 C43H74N1O7P1
747.52026 746.5 R2 (C22H31O2) - 327 746.0/327.0 M20 Plasmenyl
18:0/18:1 C41H80N1O7P1 729.56721 728.5 R2 (C18H33O2) - 281
728.0/281.0 M21 Plasmenyl 18:0/18:2 C41H78N1O7P1 727.55156 726.5 R2
(C18H31O2) - 279 726.0/279.0 M22 Plasmenyl 18:0/20:4 C43H78N1O7P1
751.55156 750.5 R2 (C20H31O2) - 303 750.6/303.2 M23 Plasmenyl
18:0/22:4 C45H82N1O7P1 779.58286 778.5 R2 (C22H35O2) - 331
778.0/331.0 M24 Plasmenyl 18:0/22:6 C45H78N1O7P1 775.55156 774.5 R2
(C22H31O2) - 327 774.0/327.0 M25 Free 22:6 C22H32O2 328.24022 327.2
(C21H31) - 283 327.2/283.0 M26 Free 20:4 C20H32O2 304.24022 303.2
(C19H31) - 259 303.2/259.5
TABLE-US-00019 TABLE 19 Clinical Data on Subject Cohorts Age MMSE
ADAS-cog Population n Mean SEM Mean SEM Mean SEM Age Ctl, 30-39,
Female 14 36.4 0.9 Age Ctl, 30-39, Male 11 35.2 1.0 Age Ctl, 40-49,
Female 44 44.8 0.5 Age Ctl, 40-49, Male 27 44.7 0.6 Age Ctl, 50-59,
Female 107 54.2 0.3 Age Ctl, 50-59, Male 59 54.1 0.4 Age Ctl,
60-69, Female 55 63.4 0.3 Age Ctl, 60-69, Male 34 64.4 0.5 Age Ctl,
70+_Female 27 79.7 1.2 Age Ctl, 70+_Male 35 75.5 0.7 Cognitive
Normal, Female 36 77.6 1.1 29.6 0.1 Cognitive Normal, Male 32 76.8
1.1 29.3 0.1 SDAT_all, Female 140 80.0 0.6 12.6 0.7 34.2 1.6
SDAT_all, Male 117 79.8 0.7 15.3 0.5 27.4 1.3 SDAT, ADAS 5-19,
Female 38 79.6 1.2 17.6 0.7 15.2 0.6 SDAT, ADAS 20-39, Female 54
78.6 1.0 16.6 0.7 27.0 0.8 SDAT, ADAS 40-70, Female 48 81.9 1.1 4.2
0.7 57.3 1.5 SDAT, ADAS 5-19, Male 40 79.0 1.1 17.3 0.7 15.3 0.5
SDAT, ADAS 20-39, Male 58 79.6 0.9 16.8 0.6 27.5 0.7 SDAT, ADAS
40-70, Male 18 82.6 2.1 6.2 1.1 53.2 2.2 Post Mortem SDAT Male 10
80.1 1.4 Post Mortem SDAT Female 10 77.6 1.5 Post Mortem Ctl,
Female 9 84.4 1.8 Post Mortem Ctl, Male 10 77.9 1.4
TABLE-US-00020 TABLE 20 Effect of Age on Serum Ethanolamine
Phospholipid Levels in Males Age Ctl, 30-39, Male Age Ctl, 40-49,
Male Age Ctl, 50-59, Male Age Ctl, 60-69, Male Age Ctl, 70+_Male
Metabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM M01
0.122 0.017 0.119 0.008 0.113 0.006 0.132 0.007 0.130 0.006 M02
0.056 0.008 0.058 0.006 0.058 0.004 0.059 0.007 0.056 0.005 M03
0.102 0.014 0.085 0.006 0.095 0.005 0.103 0.008 0.110 0.010 M04
0.026 0.004 0.027 0.003 0.025 0.002 0.027 0.003 0.027 0.003 M05
0.014 0.002 0.012 0.001 0.011 0.001 0.012 0.001 0.011 0.001 M06
0.032 0.005 0.026 0.002 0.026 0.001 0.025 0.002 0.027 0.002 M07
0.078 0.015 0.053 0.007 0.063 0.005 0.061 0.007 0.060 0.006 M08
0.010 0.002 0.007 0.001 0.007 0.001 0.007 0.001 0.006 0.001 M09
0.019 0.004 0.013 0.002 0.019 0.002 0.019 0.002 0.022 0.003 M10
0.058 0.009 0.052 0.004 0.049 0.003 0.055 0.004 0.053 0.003 M11
0.131 0.021 0.099 0.007 0.098 0.005 0.099 0.007 0.104 0.007 M12
0.244 0.037 0.186 0.015 0.210 0.012 0.213 0.017 0.202 0.016 M13
0.015 0.002 0.013 0.001 0.013 0.001 0.013 0.001 0.012 0.001 M14
0.034 0.005 0.026 0.003 0.036 0.002 0.040 0.004 0.047 0.006 M15
0.085 0.012 0.075 0.005 0.074 0.004 0.074 0.006 0.076 0.005 M16
0.291 0.043 0.220 0.016 0.244 0.013 0.223 0.018 0.246 0.018 M17
0.489 0.102 0.332 0.035 0.429 0.029 0.412 0.048 0.410 0.044 M18
0.032 0.005 0.025 0.003 0.027 0.002 0.026 0.004 0.023 0.002 M19
0.086 0.014 0.063 0.008 0.094 0.008 0.098 0.012 0.114 0.017 M20
0.086 0.013 0.070 0.006 0.068 0.004 0.069 0.006 0.071 0.006 M21
0.294 0.044 0.236 0.017 0.247 0.014 0.222 0.018 0.236 0.018 M22
1.054 0.220 0.788 0.079 0.930 0.067 0.933 0.102 0.906 0.104 M23
0.022 0.003 0.019 0.001 0.019 0.001 0.020 0.003 0.017 0.001 M24
0.077 0.014 0.063 0.008 0.086 0.007 0.088 0.010 0.103 0.016 M25
0.205 0.014 0.195 0.017 0.274 0.018 0.310 0.025 0.374 0.031 M26
0.091 0.011 0.082 0.005 0.096 0.003 0.109 0.009 0.114 0.009
TABLE-US-00021 TABLE 21 Ratio and T-test values between males of
different ages 50-59 vs. 40-49, Male 60-69 vs. 40-49, Male 70+ vs.
40-49, Male Metabolite Code Ratio ttest Ratio ttest Ratio ttest M01
0.955 6.0E-01 1.110 2.2E-01 1.097 2.4E-01 M02 0.998 9.9E-01 1.024
8.8E-01 0.965 8.1E-01 M03 1.125 2.3E-01 1.213 9.4E-02 1.296 5.0E-02
M04 0.954 7.0E-01 1.019 9.1E-01 1.015 9.3E-01 M05 0.953 6.3E-01
1.011 9.3E-01 0.979 8.5E-01 M06 1.031 7.5E-01 0.976 8.3E-01 1.054
6.2E-01 M07 1.198 2.4E-01 1.164 4.1E-01 1.133 4.8E-01 M08 1.031
8.2E-01 1.014 9.4E-01 0.907 5.4E-01 M09 1.443 2.7E-02 1.473 4.5E-02
1.701 2.1E-02 M10 0.953 5.9E-01 1.061 5.9E-01 1.013 8.9E-01 M11
0.992 9.3E-01 1.004 9.7E-01 1.058 5.6E-01 M12 1.128 2.5E-01 1.147
2.5E-01 1.085 4.8E-01 M13 1.007 9.4E-01 1.032 8.0E-01 0.921 4.4E-01
M14 1.383 1.5E-02 1.536 7.0E-03 1.802 2.8E-03 M15 0.987 8.9E-01
0.985 8.9E-01 1.020 8.5E-01 M16 1.109 2.7E-01 1.013 9.1E-01 1.118
2.9E-01 M17 1.291 5.4E-02 1.241 2.1E-01 1.235 1.9E-01 M18 1.055
6.6E-01 1.032 8.7E-01 0.904 4.9E-01 M19 1.484 1.9E-02 1.540 2.6E-02
1.789 1.9E-02 M20 0.971 7.8E-01 0.976 8.4E-01 1.014 9.0E-01 M21
1.048 6.3E-01 0.941 5.8E-01 1.000 1.0E+00 M22 1.179 2.1E-01 1.183
2.9E-01 1.149 3.9E-01 M23 1.008 9.3E-01 1.066 7.5E-01 0.900 3.6E-01
M24 1.365 5.2E-02 1.403 6.0E-02 1.642 4.7E-02 M25 1.405 9.0E-03
1.589 6.7E-04 1.912 1.7E-05 M26 1.175 1.8E-02 1.324 1.7E-02 1.389
6.3E-03
TABLE-US-00022 TABLE 22 Effect of Age on Serum Ethanolamine
Phospholipid Levels in Females Age Ctl, 30-39, Female Age Ctl,
40-49, Female Age Ctl, 50-59, Female Age Ctl, 60-69, Female Age
Ctl, 70+_Female Metabolite Code Mean SEM Mean SEM Mean SEM Mean SEM
Mean SEM M01 0.103 0.012 0.114 0.006 0.105 0.006 0.127 0.007 0.126
0.009 M02 0.047 0.006 0.069 0.006 0.058 0.004 0.058 0.004 0.056
0.004 M03 0.095 0.015 0.101 0.007 0.093 0.005 0.091 0.006 0.100
0.005 M04 0.028 0.005 0.033 0.003 0.025 0.002 0.023 0.002 0.026
0.002 M05 0.011 0.002 0.013 0.001 0.010 0.000 0.010 0.001 0.011
0.001 M06 0.031 0.005 0.031 0.002 0.025 0.001 0.023 0.001 0.025
0.002 M07 0.061 0.010 0.071 0.007 0.057 0.003 0.051 0.005 0.052
0.005 M08 0.008 0.001 0.010 0.001 0.007 0.000 0.006 0.001 0.006
0.001 M09 0.021 0.004 0.021 0.002 0.020 0.001 0.017 0.002 0.019
0.002 M10 0.050 0.007 0.059 0.003 0.049 0.002 0.052 0.003 0.051
0.003 M11 0.120 0.018 0.119 0.008 0.100 0.004 0.099 0.006 0.100
0.008 M12 0.196 0.022 0.222 0.015 0.191 0.009 0.181 0.011 0.188
0.013 M13 0.013 0.001 0.015 0.001 0.012 0.001 0.011 0.001 0.012
0.001 M14 0.040 0.007 0.043 0.004 0.042 0.003 0.041 0.003 0.049
0.004 M15 0.071 0.009 0.079 0.005 0.068 0.003 0.066 0.004 0.069
0.004 M16 0.274 0.044 0.268 0.021 0.226 0.011 0.210 0.013 0.232
0.017 M17 0.419 0.076 0.458 0.053 0.392 0.024 0.354 0.036 0.360
0.033 M18 0.028 0.004 0.032 0.003 0.025 0.002 0.019 0.002 0.023
0.003 M19 0.107 0.024 0.100 0.012 0.099 0.007 0.087 0.009 0.103
0.008 M20 0.074 0.013 0.075 0.006 0.063 0.003 0.063 0.004 0.068
0.005 M21 0.294 0.057 0.275 0.024 0.228 0.012 0.210 0.014 0.229
0.020 M22 0.938 0.170 1.025 0.108 0.853 0.055 0.759 0.068 0.837
0.096 M23 0.021 0.003 0.022 0.002 0.017 0.001 0.015 0.001 0.017
0.002 M24 0.102 0.025 0.093 0.011 0.089 0.006 0.081 0.008 0.096
0.008 M25 0.239 0.023 0.236 0.014 0.277 0.012 0.326 0.022 0.373
0.020 M26 0.091 0.012 0.093 0.006 0.093 0.003 0.094 0.004 0.107
0.007
TABLE-US-00023 TABLE 23 Ratio and T-test values between females of
different ages 50-59 vs. 40-49, Female 60-69 vs. 40-49, Female 70+
vs. 40-49, Female Metabolite Code Ratio ttest Ratio ttest Ratio
ttest M01 0.925 3.7E-01 1.118 1.8E-01 1.110 2.6E-01 M02 0.838
1.3E-01 0.835 9.5E-02 0.812 1.1E-01 M03 0.927 4.0E-01 0.906 2.9E-01
0.989 9.0E-01 M04 0.772 2.1E-02 0.709 5.1E-03 0.811 1.2E-01 M05
0.811 1.1E-02 0.798 2.0E-02 0.832 8.7E-02 M06 0.815 1.1E-02 0.731
1.5E-03 0.805 5.7E-02 M07 0.798 3.4E-02 0.718 1.9E-02 0.726 4.9E-02
M08 0.721 4.2E-03 0.569 1.3E-04 0.640 1.9E-02 M09 0.934 5.8E-01
0.784 9.4E-02 0.912 5.6E-01 M10 0.841 3.1E-02 0.882 1.3E-01 0.863
1.1E-01 M11 0.843 2.3E-02 0.832 3.5E-02 0.843 1.1E-01 M12 0.862
6.2E-02 0.815 2.5E-02 0.848 1.2E-01 M13 0.806 8.8E-03 0.737 2.0E-03
0.839 1.3E-01 M14 0.981 8.6E-01 0.953 6.9E-01 1.149 2.6E-01 M15
0.864 5.6E-02 0.841 3.7E-02 0.871 1.5E-01 M16 0.846 5.3E-02 0.786
1.5E-02 0.867 2.3E-01 M17 0.856 2.0E-01 0.773 1.0E-01 0.786 1.9E-01
M18 0.778 2.4E-02 0.594 1.8E-04 0.733 7.4E-02 M19 0.986 9.2E-01
0.872 3.8E-01 1.030 8.5E-01 M20 0.843 5.6E-02 0.842 9.5E-02 0.905
4.0E-01 M21 0.829 5.0E-02 0.763 1.8E-02 0.832 1.9E-01 M22 0.832
1.2E-01 0.741 3.3E-02 0.817 2.3E-01 M23 0.797 1.5E-02 0.663 3.3E-04
0.776 7.4E-02 M24 0.957 7.4E-01 0.871 3.6E-01 1.040 8.1E-01 M25
1.176 4.8E-02 1.381 1.7E-03 1.581 2.4E-07 M26 1.003 9.6E-01 1.021
7.9E-01 1.159 1.2E-01
TABLE-US-00024 TABLE 24 Effect of Dementia State on Serum
Ethanolamine Phospholipid Levels in Females Cognitive Normal,
SDAT_all, SDAT, ADAS SDAT, ADAS SDAT, ADAS Female Female 5-19,
Female 20-39, Female 40-70, Female Metabolite Code Mean SEM Mean
SEM Mean SEM Mean SEM Mean SEM M01 0.118 0.005 0.113 0.003 0.109
0.004 0.112 0.005 0.118 0.006 M02 0.062 0.005 0.053 0.002 0.050
0.003 0.057 0.004 0.052 0.004 M03 0.099 0.005 0.076 0.002 0.080
0.004 0.076 0.004 0.073 0.004 M04 0.026 0.002 0.025 0.001 0.024
0.002 0.027 0.002 0.024 0.003 M05 0.011 0.001 0.009 0.000 0.010
0.001 0.009 0.001 0.009 0.001 M06 0.027 0.001 0.022 0.001 0.024
0.001 0.023 0.001 0.020 0.001 M07 0.056 0.005 0.041 0.002 0.044
0.004 0.042 0.004 0.036 0.003 M08 0.006 0.001 0.005 0.000 0.005
0.000 0.006 0.001 0.005 0.000 M09 0.019 0.002 0.013 0.001 0.015
0.001 0.014 0.001 0.012 0.001 M10 0.054 0.003 0.048 0.001 0.049
0.002 0.047 0.002 0.048 0.003 M11 0.105 0.004 0.088 0.003 0.094
0.004 0.090 0.005 0.080 0.004 M12 0.196 0.012 0.153 0.005 0.166
0.009 0.152 0.010 0.142 0.010 M13 0.013 0.001 0.010 0.000 0.011
0.000 0.010 0.001 0.010 0.001 M14 0.046 0.003 0.035 0.002 0.040
0.003 0.034 0.002 0.033 0.003 M15 0.073 0.004 0.059 0.002 0.061
0.003 0.061 0.003 0.056 0.003 M16 0.250 0.013 0.192 0.007 0.214
0.013 0.201 0.012 0.164 0.008 M17 0.408 0.033 0.288 0.015 0.317
0.026 0.304 0.026 0.247 0.021 M18 0.024 0.002 0.018 0.001 0.019
0.001 0.019 0.002 0.016 0.001 M19 0.103 0.008 0.071 0.004 0.079
0.007 0.072 0.006 0.063 0.006 M20 0.077 0.004 0.059 0.002 0.064
0.004 0.060 0.004 0.054 0.003 M21 0.265 0.015 0.195 0.008 0.219
0.015 0.205 0.015 0.165 0.011 M22 0.933 0.077 0.702 0.036 0.753
0.061 0.738 0.066 0.620 0.057 M23 0.018 0.001 0.014 0.001 0.015
0.001 0.014 0.001 0.013 0.001 M24 0.103 0.009 0.073 0.004 0.079
0.008 0.073 0.007 0.067 0.008 M25 0.241 0.013 0.211 0.007 0.218
0.013 0.212 0.014 0.204 0.011 M26 0.069 0.003 0.073 0.002 0.076
0.003 0.072 0.004 0.072 0.003
TABLE-US-00025 TABLE 25 Ratio and T-test values between females of
various levels of dementia AD, All to CN, Female ADAS 5-19 to CN,
Female ADAS 20-39 to CN, Female ADAS 40-70 to CN, Female Metabolite
Code Ratio ttest Ratio ttest Ratio ttest Ratio ttest M01 0.963
5.2E-01 0.929 2.1E-01 0.951 4.4E-01 1.004 9.6E-01 M02 0.856 9.0E-02
0.806 4.5E-02 0.912 3.9E-01 0.833 1.2E-01 M03 0.772 5.2E-05 0.814
6.0E-03 0.775 8.0E-04 0.737 1.8E-04 M04 0.963 7.5E-01 0.912 3.8E-01
1.027 8.3E-01 0.932 6.2E-01 M05 0.893 1.1E-01 0.925 3.4E-01 0.895
2.1E-01 0.867 8.4E-02 M06 0.843 1.4E-02 0.910 2.2E-01 0.869 9.8E-02
0.761 5.1E-04 M07 0.732 1.8E-03 0.798 5.9E-02 0.753 2.0E-02 0.656
1.1E-03 M08 0.821 5.0E-02 0.849 1.4E-01 0.889 3.7E-01 0.722 4.7E-03
M09 0.696 2.6E-04 0.777 3.9E-02 0.702 2.9E-03 0.624 4.3E-04 M10
0.877 3.1E-02 0.895 8.6E-02 0.860 4.1E-02 0.883 9.8E-02 M11 0.832
2.5E-03 0.894 7.5E-02 0.850 2.7E-02 0.762 1.4E-04 M12 0.778 5.3E-04
0.847 4.2E-02 0.776 4.7E-03 0.726 5.8E-04 M13 0.800 8.5E-04 0.834
2.5E-02 0.794 8.6E-03 0.780 3.2E-03 M14 0.772 4.8E-03 0.869 1.9E-01
0.746 3.0E-03 0.724 5.6E-03 M15 0.811 8.5E-04 0.835 1.5E-02 0.831
2.3E-02 0.770 4.4E-04 M16 0.765 7.5E-05 0.853 4.3E-02 0.801 6.9E-03
0.656 6.4E-08 M17 0.705 3.5E-04 0.776 3.2E-02 0.745 1.4E-02 0.605
4.3E-05 M18 0.754 3.4E-03 0.804 5.9E-02 0.792 6.8E-02 0.673 7.9E-04
M19 0.688 1.8E-04 0.768 2.5E-02 0.699 2.9E-03 0.612 7.0E-05 M20
0.768 4.9E-04 0.828 3.0E-02 0.782 1.1E-02 0.703 4.8E-05 M21 0.737
1.9E-04 0.826 3.7E-02 0.776 1.1E-02 0.624 6.8E-07 M22 0.752 5.2E-03
0.807 7.0E-02 0.790 6.1E-02 0.665 1.3E-03 M23 0.764 2.5E-03 0.809
3.2E-02 0.789 4.1E-02 0.699 7.8E-04 M24 0.708 2.5E-03 0.768 4.5E-02
0.713 9.3E-03 0.654 4.2E-03 M25 0.876 6.6E-02 0.907 2.3E-01 0.880
1.5E-01 0.847 3.5E-02 M26 1.056 3.8E-01 1.092 1.8E-01 1.043 5.8E-01
1.041 5.5E-01
TABLE-US-00026 TABLE 26 Ratio and T-test values between females of
various levels of dementia ADAS 20-39 to 5-19, Female ADAS 40-70 to
5-19, Female ADAS 40-70 to 20-39, Female Metabolite Code Ratio
ttest Ratio ttest Ratio ttest M01 1.023 7.2E-01 1.080 2.8E-01 1.056
4.4E-01 M02 1.132 2.3E-01 1.034 7.6E-01 0.914 4.1E-01 M03 0.951
5.1E-01 0.906 2.2E-01 0.952 5.3E-01 M04 1.126 3.6E-01 1.022 8.8E-01
0.907 5.0E-01 M05 0.967 7.0E-01 0.936 4.2E-01 0.968 7.1E-01 M06
0.955 5.9E-01 0.837 2.5E-02 0.876 1.2E-01 M07 0.944 6.4E-01 0.822
1.2E-01 0.871 2.8E-01 M08 1.046 7.2E-01 0.850 1.1E-01 0.812 1.1E-01
M09 0.903 3.9E-01 0.802 9.5E-02 0.889 3.5E-01 M10 0.961 5.8E-01
0.987 8.6E-01 1.027 7.2E-01 M11 0.951 5.1E-01 0.853 2.8E-02 0.896
1.6E-01 M12 0.917 3.1E-01 0.858 7.7E-02 0.935 4.7E-01 M13 0.953
5.4E-01 0.936 3.6E-01 0.982 8.2E-01 M14 0.858 1.6E-01 0.833 1.5E-01
0.971 8.0E-01 M15 0.996 9.6E-01 0.922 2.3E-01 0.926 3.1E-01 M16
0.940 4.7E-01 0.769 9.7E-04 0.819 1.6E-02 M17 0.960 7.4E-01 0.779
4.0E-02 0.812 9.8E-02 M18 0.985 9.0E-01 0.837 7.6E-02 0.850 1.8E-01
M19 0.911 4.7E-01 0.798 8.5E-02 0.875 3.1E-01 M20 0.945 5.7E-01
0.849 5.8E-02 0.898 2.7E-01 M21 0.939 5.5E-01 0.755 4.6E-03 0.804
4.1E-02 M22 0.979 8.7E-01 0.824 1.2E-01 0.841 1.9E-01 M23 0.975
8.3E-01 0.864 1.4E-01 0.886 3.0E-01 M24 0.928 6.0E-01 0.852 3.2E-01
0.918 5.8E-01 M25 0.971 7.5E-01 0.934 4.0E-01 0.962 6.6E-01 M26
0.955 5.2E-01 0.954 4.5E-01 0.998 9.8E-01
TABLE-US-00027 TABLE 27 Average Serum Ethanolamine Phospholipid
Levels in Males of Different Levels of Dementia Severity Cognitive
Normal, Male SDAT_all, Male SDAT, ADAS 5-19, Male SDAT, ADAS 20-39,
Male SDAT, ADAS 40-70, Male Metabolite Code Mean SEM Mean SEM Mean
SEM Mean SEM Mean SEM M01 0.116 0.006 0.119 0.004 0.117 0.006 0.117
0.006 0.117 0.011 M02 0.066 0.006 0.056 0.004 0.060 0.009 0.054
0.004 0.046 0.006 M03 0.109 0.007 0.084 0.004 0.091 0.006 0.080
0.005 0.072 0.007 M04 0.030 0.002 0.026 0.002 0.025 0.004 0.027
0.002 0.022 0.003 M05 0.012 0.001 0.010 0.000 0.011 0.001 0.010
0.001 0.010 0.001 M06 0.028 0.002 0.025 0.001 0.024 0.002 0.025
0.002 0.024 0.002 M07 0.062 0.005 0.044 0.002 0.047 0.004 0.044
0.003 0.036 0.004 M08 0.007 0.001 0.005 0.000 0.006 0.001 0.005
0.000 0.004 0.001 M09 0.022 0.002 0.015 0.001 0.016 0.002 0.015
0.001 0.011 0.001 M10 0.054 0.002 0.052 0.002 0.053 0.003 0.051
0.002 0.047 0.005 M11 0.110 0.005 0.095 0.004 0.095 0.007 0.095
0.006 0.088 0.009 M12 0.205 0.011 0.162 0.006 0.170 0.011 0.160
0.008 0.141 0.013 M13 0.013 0.001 0.011 0.000 0.011 0.001 0.010
0.001 0.010 0.001 M14 0.051 0.004 0.036 0.002 0.041 0.004 0.034
0.002 0.028 0.003 M15 0.076 0.004 0.064 0.002 0.067 0.005 0.062
0.003 0.060 0.006 M16 0.266 0.015 0.207 0.009 0.213 0.015 0.202
0.012 0.191 0.018 M17 0.470 0.038 0.302 0.015 0.338 0.031 0.292
0.020 0.242 0.026 M18 0.026 0.003 0.019 0.001 0.020 0.002 0.018
0.001 0.016 0.002 M19 0.127 0.013 0.078 0.004 0.088 0.009 0.076
0.006 0.056 0.006 M20 0.078 0.005 0.064 0.003 0.069 0.005 0.061
0.003 0.059 0.007 M21 0.265 0.017 0.217 0.010 0.226 0.019 0.211
0.014 0.201 0.023 M22 1.040 0.087 0.736 0.037 0.789 0.071 0.723
0.052 0.624 0.067 M23 0.018 0.001 0.015 0.001 0.016 0.001 0.014
0.001 0.013 0.001 M24 0.116 0.012 0.079 0.005 0.090 0.010 0.075
0.006 0.061 0.008 M25 0.240 0.017 0.218 0.009 0.249 0.018 0.208
0.010 0.183 0.017 M26 0.072 0.003 0.070 0.002 0.074 0.004 0.070
0.003 0.065 0.006
TABLE-US-00028 TABLE 28 Ratio and T-test Values Between Males of
Various Levels of Dementia AD, All to CN, Male ADAS 5-19 to CN,
Male ADAS 20-39 to CN, Male ADAS 40-70 to CN, Male Metabolite Code
Ratio ttest Ratio ttest Ratio ttest Ratio ttest M01 1.028 7.1E-01
1.011 8.9E-01 1.016 8.3E-01 1.014 8.9E-01 M02 0.845 2.1E-01 0.898
5.5E-01 0.816 7.5E-02 0.690 2.4E-02 M03 0.769 1.6E-03 0.836 6.9E-02
0.735 6.4E-04 0.655 1.4E-03 M04 0.882 3.9E-01 0.836 2.9E-01 0.896
4.1E-01 0.726 2.7E-02 M05 0.882 1.3E-01 0.896 2.7E-01 0.865 9.1E-02
0.836 1.5E-01 M06 0.877 1.3E-01 0.855 9.1E-02 0.876 1.6E-01 0.856
1.4E-01 M07 0.709 3.9E-04 0.753 1.8E-02 0.704 1.6E-03 0.589 1.1E-03
M08 0.759 1.9E-02 0.806 1.8E-01 0.755 2.1E-02 0.629 1.5E-02 M09
0.658 2.7E-04 0.731 2.7E-02 0.654 1.4E-03 0.472 2.2E-04 M10 0.960
6.0E-01 0.978 7.9E-01 0.942 4.0E-01 0.877 1.7E-01 M11 0.866 9.3E-02
0.861 1.1E-01 0.861 8.1E-02 0.798 2.6E-02 M12 0.793 1.8E-03 0.831
3.9E-02 0.783 2.2E-03 0.686 8.3E-04 M13 0.850 2.7E-02 0.890 1.8E-01
0.827 1.3E-02 0.786 1.8E-02 M14 0.704 3.7E-04 0.800 6.9E-02 0.666
1.1E-04 0.553 2.3E-04 M15 0.842 2.2E-02 0.883 1.8E-01 0.810 3.0E-03
0.784 1.9E-02 M16 0.778 1.6E-03 0.802 1.7E-02 0.759 1.2E-03 0.718
3.2E-03 M17 0.644 4.5E-06 0.718 7.5E-03 0.621 1.3E-05 0.515 1.0E-04
M18 0.708 1.6E-03 0.758 6.0E-02 0.689 2.0E-03 0.613 7.7E-03 M19
0.611 1.0E-05 0.688 1.1E-02 0.596 6.1E-05 0.442 2.0E-04 M20 0.826
2.3E-02 0.892 2.7E-01 0.786 6.1E-03 0.762 3.9E-02 M21 0.818 2.7E-02
0.852 1.3E-01 0.796 2.0E-02 0.757 2.9E-02 M22 0.708 4.4E-04 0.758
2.7E-02 0.695 1.3E-03 0.600 1.9E-03 M23 0.801 1.1E-02 0.857 1.6E-01
0.776 4.0E-03 0.723 8.6E-03 M24 0.680 1.1E-03 0.777 9.8E-02 0.647
1.3E-03 0.524 2.8E-03 M25 0.909 2.4E-01 1.035 7.4E-01 0.865 8.0E-02
0.760 3.0E-02 M26 0.978 7.4E-01 1.023 7.5E-01 0.968 6.5E-01 0.907
2.9E-01
TABLE-US-00029 TABLE 29 Ratio and T-test Values Between Males of
Various Levels of Dementia ADAS 20-39 to 5-19, Male ADAS 40-70 to
5-19, Male ADAS 40-70 to 20-39, Male Metabolite Code Ratio ttest
Ratio ttest Ratio ttest M01 1.005 9.4E-01 1.003 9.8E-01 0.998
9.8E-01 M02 0.908 5.3E-01 0.768 3.2E-01 0.846 2.8E-01 M03 0.880
1.5E-01 0.784 7.0E-02 0.891 3.4E-01 M04 1.072 6.8E-01 0.868 5.8E-01
0.810 2.8E-01 M05 0.966 7.0E-01 0.933 6.1E-01 0.966 7.7E-01 M06
1.024 8.1E-01 1.001 9.9E-01 0.978 8.6E-01 M07 0.935 5.5E-01 0.783
1.3E-01 0.837 2.5E-01 M08 0.936 6.3E-01 0.780 2.7E-01 0.833 2.4E-01
M09 0.895 4.2E-01 0.646 3.3E-02 0.721 1.0E-01 M10 0.963 6.3E-01
0.897 3.7E-01 0.932 5.0E-01 M11 1.000 1.0E+00 0.927 5.7E-01 0.927
5.4E-01 M12 0.942 4.8E-01 0.826 1.3E-01 0.877 2.4E-01 M13 0.930
3.8E-01 0.883 3.2E-01 0.950 6.3E-01 M14 0.832 1.1E-01 0.691 4.5E-02
0.830 2.0E-01 M15 0.917 3.0E-01 0.888 3.7E-01 0.968 7.4E-01 M16
0.947 5.4E-01 0.895 3.9E-01 0.946 6.4E-01 M17 0.865 1.9E-01 0.716
5.6E-02 0.828 1.9E-01 M18 0.909 4.5E-01 0.809 2.8E-01 0.890 4.4E-01
M19 0.866 2.5E-01 0.642 3.1E-02 0.741 7.4E-02 M20 0.880 1.6E-01
0.854 2.7E-01 0.970 8.0E-01 M21 0.934 5.2E-01 0.889 4.4E-01 0.952
7.2E-01 M22 0.917 4.5E-01 0.791 1.6E-01 0.863 3.3E-01 M23 0.906
3.3E-01 0.844 2.8E-01 0.931 5.6E-01 M24 0.833 1.7E-01 0.674 5.9E-02
0.810 2.3E-01 M25 0.836 3.7E-02 0.735 2.8E-02 0.879 2.1E-01 M26
0.946 4.3E-01 0.887 2.3E-01 0.937 5.2E-01
TABLE-US-00030 TABLE 30 Effect of Pathology State on Serum
Ethanolamine Phospholipid Levels in Males Post Mortem Ctl, Male
Post Mortem SDAT Male SDAT vs Control Metabolite Code Mean SEM Mean
SEM Ratio ttest M01 0.127 0.017 0.089 0.013 0.702 0.091 M02 0.046
0.006 0.026 0.005 0.568 0.022 M03 0.059 0.006 0.036 0.006 0.610
0.014 M04 0.017 0.004 0.007 0.002 0.420 0.024 M05 0.006 0.001 0.004
0.000 0.479 0.019 M06 0.009 0.001 0.006 0.001 0.475 0.005 M07 0.012
0.003 0.009 0.001 0.451 0.033 M08 0.003 0.001 0.002 0.000 0.410
0.015 M09 0.006 0.002 0.003 0.001 0.269 0.048 M10 0.041 0.005 0.036
0.006 0.608 0.019 M11 0.052 0.006 0.041 0.006 0.474 0.001 M12 0.094
0.013 0.084 0.012 0.587 0.024 M13 0.009 0.001 0.008 0.001 0.576
0.008 M14 0.025 0.005 0.021 0.007 0.429 0.009 M15 0.045 0.005 0.029
0.004 0.648 0.026 M16 0.092 0.012 0.053 0.007 0.570 0.012 M17 0.097
0.021 0.047 0.007 0.489 0.036 M18 0.010 0.001 0.005 0.001 0.521
0.004 M19 0.032 0.006 0.014 0.002 0.452 0.011 M20 0.031 0.004 0.017
0.002 0.542 0.006 M21 0.072 0.011 0.033 0.006 0.464 0.006 M22 0.217
0.040 0.106 0.017 0.486 0.020 M23 0.009 0.001 0.005 0.001 0.565
0.005 M24 0.029 0.004 0.013 0.003 0.448 0.007 M25 0.238 0.023 0.180
0.026 0.757 0.114 M26 0.073 0.008 0.050 0.006 0.684 0.034
TABLE-US-00031 TABLE 31 Effect of Pathology State on Serum
Ethanolamine Phospholipid Levels in Females Post Mortem Ctl, Female
Post Mortem SDAT Female Autopsy AD vs. Control, Female Metabolite
Code Mean SEM Mean SEM Ratio ttest M01 0.179 0.050 0.124 0.016
0.697 0.300 M02 0.062 0.022 0.048 0.011 0.773 0.557 M03 0.070 0.019
0.043 0.005 0.619 0.178 M04 0.016 0.005 0.012 0.004 0.775 0.563 M05
0.007 0.002 0.003 0.001 0.508 0.095 M06 0.011 0.003 0.004 0.001
0.531 0.060 M07 0.016 0.003 0.005 0.001 0.530 0.037 M08 0.003 0.001
0.001 0.000 0.684 0.258 M09 0.005 0.001 0.002 0.000 0.566 0.047 M10
0.053 0.015 0.025 0.004 0.672 0.293 M11 0.064 0.013 0.025 0.004
0.650 0.132 M12 0.114 0.021 0.055 0.008 0.742 0.241 M13 0.010 0.002
0.005 0.001 0.865 0.537 M14 0.023 0.004 0.011 0.002 0.904 0.792 M15
0.060 0.017 0.034 0.004 0.577 0.151 M16 0.107 0.021 0.064 0.008
0.594 0.061 M17 0.113 0.024 0.067 0.009 0.592 0.079 M18 0.014 0.004
0.008 0.001 0.579 0.186 M19 0.035 0.007 0.022 0.004 0.635 0.132 M20
0.050 0.014 0.021 0.002 0.410 0.042 M21 0.100 0.020 0.041 0.007
0.414 0.010 M22 0.283 0.058 0.168 0.024 0.593 0.071 M23 0.012 0.003
0.007 0.001 0.560 0.112 M24 0.035 0.007 0.018 0.002 0.496 0.025 M25
0.189 0.015 0.198 0.022 1.051 0.727 M26 0.054 0.008 0.061 0.006
1.123 0.496
TABLE-US-00032 TABLE 32 Effect of Age on Ethanolamine Phospholipid
Ratios to M01 in Males Age Ctl, 30-39, Male Age Ctl, 40-49, Male
Age Ctl, 50-59, Male Age Ctl, 60-69, Male Age Ctl, 70+_Male
Metabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM M01
1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000 M02
0.476 0.042 0.477 0.034 0.517 0.027 0.431 0.031 0.420 0.028 M03
0.863 0.063 0.751 0.049 0.864 0.030 0.792 0.051 0.848 0.071 M04
0.224 0.025 0.229 0.026 0.235 0.013 0.203 0.019 0.204 0.017 M05
0.113 0.008 0.102 0.007 0.101 0.004 0.091 0.005 0.088 0.005 M06
0.263 0.029 0.234 0.021 0.243 0.012 0.199 0.016 0.210 0.012 M07
0.683 0.136 0.493 0.070 0.581 0.041 0.479 0.052 0.457 0.039 M08
0.079 0.010 0.064 0.007 0.067 0.004 0.054 0.005 0.049 0.004 M09
0.161 0.025 0.124 0.017 0.173 0.013 0.154 0.018 0.172 0.024 M10
0.464 0.025 0.454 0.022 0.445 0.012 0.419 0.018 0.407 0.016 M11
1.063 0.111 0.899 0.069 0.908 0.037 0.789 0.052 0.811 0.038 M12
2.068 0.251 1.728 0.168 1.942 0.099 1.664 0.111 1.552 0.087 M13
0.126 0.009 0.117 0.009 0.121 0.005 0.102 0.006 0.092 0.005 M14
0.294 0.033 0.242 0.029 0.332 0.020 0.316 0.029 0.363 0.041 M15
0.699 0.034 0.658 0.041 0.675 0.023 0.567 0.031 0.590 0.029 M16
2.429 0.285 2.020 0.175 2.284 0.118 1.778 0.145 1.922 0.112 M17
4.127 0.757 3.164 0.418 3.980 0.257 3.167 0.320 3.178 0.294 M18
0.258 0.025 0.229 0.025 0.244 0.014 0.195 0.021 0.174 0.014 M19
0.725 0.101 0.607 0.094 0.865 0.063 0.769 0.085 0.881 0.131 M20
0.691 0.049 0.616 0.043 0.619 0.030 0.529 0.034 0.553 0.034 M21
2.395 0.215 2.163 0.189 2.287 0.122 1.773 0.148 1.840 0.108 M22
8.811 1.529 7.346 0.883 8.537 0.549 7.124 0.644 6.920 0.635 M23
0.189 0.019 0.175 0.016 0.178 0.009 0.150 0.016 0.133 0.008 M24
0.649 0.086 0.606 0.098 0.787 0.057 0.700 0.072 0.799 0.122 M25
2.182 0.444 1.975 0.265 2.747 0.211 2.608 0.251 3.066 0.284
TABLE-US-00033 TABLE 33 Ratio and T-test values of Ethanolamine
Phospholipid Ratios to M01 between males of different ages 50-59
vs. 40-49, Male 60-69 vs. 40-49, Male 70+ vs. 40-49, Male
Metabolite Code Ratio ttest Ratio ttest Ratio ttest M01 1.000
#DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 1.082 3.9E-01 0.903 3.2E-01
0.879 1.9E-01 M03 1.149 4.5E-02 1.054 5.8E-01 1.128 3.0E-01 M04
1.023 8.4E-01 0.883 4.0E-01 0.889 3.9E-01 M05 0.984 8.3E-01 0.884
1.6E-01 0.863 8.5E-02 M06 1.041 6.8E-01 0.851 1.9E-01 0.898 3.1E-01
M07 1.177 2.6E-01 0.970 8.6E-01 0.927 6.4E-01 M08 1.053 6.7E-01
0.847 2.6E-01 0.765 5.4E-02 M09 1.404 2.5E-02 1.243 2.4E-01 1.391
1.3E-01 M10 0.981 7.1E-01 0.924 2.3E-01 0.896 7.6E-02 M11 1.010
9.0E-01 0.878 2.0E-01 0.903 2.5E-01 M12 1.124 2.5E-01 0.963 7.4E-01
0.898 3.3E-01 M13 1.035 6.7E-01 0.873 1.7E-01 0.787 1.3E-02 M14
1.372 1.2E-02 1.306 7.7E-02 1.500 2.7E-02 M15 1.026 7.0E-01 0.862
7.8E-02 0.896 1.7E-01 M16 1.131 2.1E-01 0.880 2.9E-01 0.951 6.2E-01
M17 1.258 8.8E-02 1.001 1.0E+00 1.004 9.8E-01 M18 1.067 5.6E-01
0.851 3.0E-01 0.761 4.9E-02 M19 1.426 2.4E-02 1.266 2.1E-01 1.451
1.1E-01 M20 1.004 9.6E-01 0.858 1.1E-01 0.898 2.5E-01 M21 1.057
5.8E-01 0.820 1.0E-01 0.851 1.2E-01 M22 1.162 2.4E-01 0.970 8.4E-01
0.942 6.9E-01 M23 1.016 8.7E-01 0.858 2.8E-01 0.759 1.5E-02 M24
1.300 9.4E-02 1.156 4.3E-01 1.319 2.4E-01 M25 1.391 3.5E-02 1.321
9.0E-02 1.553 8.2E-03
TABLE-US-00034 TABLE 34 Effect of Age on Ethanolamine Phospholipid
Ratios to M01 in Females Age Ctl, 30-39, Female Age Ctl, 40-49,
Female Age Ctl, 50-59, Female Age Ctl, 60-69, Female Age Ctl,
70+_Female Metabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean
SEM M01 1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000
M02 0.474 0.037 0.597 0.028 0.551 0.020 0.460 0.018 0.455 0.023 M03
0.931 0.089 0.911 0.043 0.926 0.030 0.750 0.034 0.838 0.048 M04
0.279 0.036 0.283 0.017 0.241 0.011 0.184 0.010 0.221 0.020 M05
0.112 0.009 0.118 0.006 0.105 0.003 0.085 0.005 0.089 0.006 M06
0.311 0.044 0.291 0.019 0.261 0.010 0.193 0.012 0.213 0.017 M07
0.618 0.090 0.667 0.056 0.582 0.030 0.435 0.039 0.430 0.041 M08
0.079 0.008 0.089 0.007 0.070 0.003 0.047 0.004 0.052 0.007 M09
0.204 0.036 0.194 0.017 0.197 0.011 0.139 0.012 0.163 0.015 M10
0.491 0.027 0.535 0.024 0.490 0.010 0.424 0.015 0.422 0.022 M11
1.220 0.159 1.092 0.060 1.021 0.031 0.823 0.037 0.840 0.058 M12
1.979 0.150 2.044 0.117 1.952 0.063 1.534 0.086 1.564 0.100 M13
0.129 0.008 0.136 0.007 0.122 0.004 0.092 0.005 0.104 0.010 M14
0.379 0.047 0.381 0.024 0.414 0.018 0.333 0.021 0.412 0.033 M15
0.706 0.044 0.720 0.034 0.699 0.019 0.560 0.024 0.578 0.033 M16
2.739 0.333 2.483 0.161 2.345 0.091 1.783 0.096 1.964 0.136 M17
4.055 0.499 4.149 0.401 3.987 0.209 2.963 0.275 3.010 0.268 M18
0.282 0.022 0.289 0.022 0.245 0.010 0.160 0.012 0.191 0.024 M19
1.009 0.175 0.902 0.085 0.974 0.056 0.716 0.061 0.877 0.077 M20
0.708 0.059 0.682 0.045 0.637 0.021 0.524 0.029 0.565 0.039 M21
2.885 0.411 2.537 0.187 2.331 0.092 1.757 0.105 1.935 0.158 M22
8.971 0.932 9.244 0.790 8.625 0.450 6.263 0.484 6.923 0.683 M23
0.198 0.011 0.202 0.014 0.177 0.006 0.124 0.008 0.140 0.014 M24
0.955 0.174 0.834 0.078 0.882 0.049 0.661 0.054 0.814 0.071 M25
2.505 0.217 2.286 0.164 3.219 0.181 2.995 0.239 3.280 0.248
TABLE-US-00035 TABLE 35 Ratio and T-test values of Ethanolamine
Phospholipid Ratios to M01 between females of different ages 50-59
vs. 40-49, Female 60-69 vs. 40-49, Female 70+ vs. 40-49, Female
Metabolite Code Ratio ttest Ratio ttest Ratio ttest M01 1.000
#DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 0.923 2.1E-01 0.771 5.7E-05
0.763 8.4E-04 M03 1.017 7.8E-01 0.823 3.7E-03 0.920 2.8E-01 M04
0.850 3.5E-02 0.652 1.1E-06 0.782 2.3E-02 M05 0.892 4.7E-02 0.721
5.0E-05 0.758 3.7E-03 M06 0.896 1.3E-01 0.662 1.3E-05 0.733 6.5E-03
M07 0.873 1.5E-01 0.652 7.2E-04 0.644 3.5E-03 M08 0.783 4.7E-03
0.533 6.9E-07 0.583 9.9E-04 M09 1.016 8.8E-01 0.717 7.8E-03 0.841
2.2E-01 M10 0.917 4.8E-02 0.792 7.9E-05 0.789 2.0E-03 M11 0.935
2.5E-01 0.754 1.3E-04 0.770 5.9E-03 M12 0.955 4.6E-01 0.751 5.1E-04
0.765 5.8E-03 M13 0.900 7.0E-02 0.680 4.8E-06 0.764 9.1E-03 M14
1.088 3.1E-01 0.876 1.4E-01 1.083 4.3E-01 M15 0.970 5.6E-01 0.778
1.8E-04 0.802 6.7E-03 M16 0.944 4.3E-01 0.718 1.8E-04 0.791 2.8E-02
M17 0.961 7.0E-01 0.714 1.4E-02 0.726 4.4E-02 M18 0.847 4.1E-02
0.552 6.0E-07 0.660 5.6E-03 M19 1.079 4.9E-01 0.794 7.2E-02 0.972
8.4E-01 M20 0.935 3.1E-01 0.769 2.9E-03 0.828 7.5E-02 M21 0.919
2.7E-01 0.693 2.4E-04 0.763 2.9E-02 M22 0.933 4.8E-01 0.678 1.2E-03
0.749 4.6E-02 M23 0.876 5.5E-02 0.614 1.3E-06 0.696 3.9E-03 M24
1.058 6.0E-01 0.793 6.5E-02 0.976 8.6E-01 M25 1.408 2.4E-03 1.310
2.2E-02 1.435 8.6E-04
TABLE-US-00036 TABLE 36 Average Serum Ethanolamine Phospholipid
Ratios to M01 in Males of Different Levels of Dementia Severity
Cognitive Normal, Male SDAT_all, Male SDAT, ADAS 5-19, Male SDAT,
ADAS 20-39, Male SDAT, ADAS 40-70, Male Metabolite Code Mean SEM
Mean SEM Mean SEM Mean SEM Mean SEM M01 1.000 0.000 1.000 0.000
1.000 0.000 1.000 0.000 1.000 0.000 M02 0.564 0.031 0.453 0.015
0.475 0.032 0.455 0.017 0.383 0.022 M03 0.960 0.050 0.712 0.018
0.779 0.037 0.694 0.024 0.617 0.027 M04 0.262 0.015 0.210 0.009
0.203 0.017 0.219 0.013 0.183 0.014 M05 0.106 0.006 0.089 0.002
0.091 0.004 0.089 0.003 0.085 0.004 M06 0.255 0.014 0.213 0.007
0.211 0.012 0.214 0.010 0.211 0.012 M07 0.554 0.041 0.373 0.014
0.396 0.025 0.375 0.020 0.321 0.028 M08 0.062 0.005 0.044 0.002
0.047 0.004 0.045 0.003 0.038 0.003 M09 0.195 0.015 0.126 0.006
0.139 0.012 0.127 0.009 0.093 0.008 M10 0.483 0.021 0.440 0.009
0.450 0.016 0.441 0.013 0.408 0.016 M11 0.985 0.043 0.817 0.025
0.828 0.051 0.824 0.035 0.761 0.036 M12 1.843 0.088 1.399 0.036
1.460 0.059 1.406 0.053 1.252 0.090 M13 0.114 0.006 0.092 0.002
0.096 0.004 0.091 0.003 0.088 0.006 M14 0.442 0.023 0.310 0.013
0.348 0.028 0.301 0.015 0.255 0.025 M15 0.682 0.031 0.548 0.012
0.571 0.022 0.541 0.016 0.520 0.024 M16 2.398 0.128 1.790 0.057
1.856 0.105 1.777 0.082 1.687 0.115 M17 4.203 0.304 2.569 0.105
2.853 0.205 2.501 0.135 2.187 0.243 M18 0.232 0.017 0.156 0.006
0.166 0.012 0.155 0.009 0.140 0.010 M19 1.103 0.092 0.663 0.032
0.740 0.066 0.660 0.041 0.503 0.045 M20 0.692 0.037 0.548 0.016
0.593 0.030 0.532 0.021 0.509 0.034 M21 2.377 0.126 1.857 0.066
1.951 0.128 1.829 0.090 1.754 0.143 M22 9.309 0.674 6.230 0.231
6.651 0.434 6.157 0.305 5.616 0.592 M23 0.164 0.009 0.125 0.004
0.133 0.008 0.124 0.006 0.114 0.008 M24 1.010 0.088 0.672 0.034
0.770 0.076 0.647 0.039 0.539 0.053 M25 2.160 0.133 2.085 0.115
2.215 0.145 2.102 0.197 1.811 0.222
TABLE-US-00037 TABLE 37 Ratio and T-test Values of Ethanolamine
Phospholipid Ratios to M01 between Males of Various Levels of
Dementia AD, All to CN, Male ADAS 5-19 to CN, Male ADAS 20-39 to
CN, Male ADAS 40-70 to CN, Male Metabolite Code Ratio ttest Ratio
ttest Ratio ttest Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000
#DIV/0! 1.000 #DIV/0! M02 0.803 7.2E-04 0.842 5.2E-02 0.807 1.2E-03
0.680 1.8E-04 M03 0.742 6.4E-08 0.812 3.8E-03 0.724 4.5E-07 0.643
9.8E-06 M04 0.802 8.4E-03 0.774 1.6E-02 0.836 4.1E-02 0.700 1.3E-03
M05 0.840 9.2E-04 0.858 2.9E-02 0.841 5.5E-03 0.798 1.2E-02 M06
0.833 5.0E-03 0.827 2.0E-02 0.840 2.2E-02 0.828 4.7E-02 M07 0.673
4.7E-07 0.715 1.1E-03 0.676 3.1E-05 0.580 2.4E-04 M08 0.713 1.1E-04
0.747 1.3E-02 0.727 1.5E-03 0.602 8.4E-04 M09 0.645 2.8E-06 0.713
4.4E-03 0.654 7.5E-05 0.476 1.6E-05 M10 0.910 3.4E-02 0.932 2.1E-01
0.913 8.0E-02 0.843 1.8E-02 M11 0.830 1.9E-03 0.841 2.5E-02 0.837
5.8E-03 0.773 9.3E-04 M12 0.759 3.4E-07 0.792 3.9E-04 0.763 1.9E-05
0.679 7.0E-05 M13 0.811 1.0E-04 0.846 1.2E-02 0.802 4.0E-04 0.772
4.7E-03 M14 0.701 4.1E-06 0.787 1.4E-02 0.680 1.1E-06 0.576 4.9E-06
M15 0.803 2.7E-06 0.837 3.8E-03 0.792 2.4E-05 0.762 8.6E-04 M16
0.747 4.3E-06 0.774 1.5E-03 0.741 4.8E-05 0.704 5.4E-04 M17 0.611
2.1E-09 0.679 3.1E-04 0.595 7.2E-08 0.520 4.0E-05 M18 0.672 9.8E-07
0.714 1.7E-03 0.666 2.3E-05 0.603 3.3E-04 M19 0.601 6.5E-08 0.672
1.7E-03 0.599 2.1E-06 0.456 2.2E-05 M20 0.793 9.1E-05 0.858 3.8E-02
0.769 1.0E-04 0.736 1.8E-03 M21 0.782 3.7E-04 0.821 2.2E-02 0.770
5.6E-04 0.738 3.0E-03 M22 0.669 2.0E-07 0.714 9.9E-04 0.661 4.8E-06
0.603 6.0E-04 M23 0.763 1.2E-04 0.810 1.5E-02 0.754 4.8E-04 0.696
6.5E-04 M24 0.665 3.5E-05 0.763 4.2E-02 0.641 3.5E-05 0.533 4.0E-04
M25 0.965 7.5E-01 1.025 7.9E-01 0.973 8.4E-01 0.838 1.6E-01
TABLE-US-00038 TABLE 38 Ratio and T-test Values of Ethanolamine
Phospholipid Ratios to M01 Between Males of Various Levels of
Dementia ADAS 20-39 to 5-19, Male ADAS 40-70 to 5-19, Male ADAS
40-70 to 20-39, Male Metabolite Code Ratio ttest Ratio ttest Ratio
ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 0.958
5.5E-01 0.807 7.1E-02 0.843 3.5E-02 M03 0.891 4.6E-02 0.792 6.9E-03
0.888 9.3E-02 M04 1.080 4.5E-01 0.903 4.8E-01 0.837 1.4E-01 M05
0.980 7.2E-01 0.930 3.2E-01 0.948 4.4E-01 M06 1.016 8.4E-01 1.002
9.8E-01 0.987 8.9E-01 M07 0.946 5.1E-01 0.811 8.4E-02 0.857 1.8E-01
M08 0.973 7.8E-01 0.806 1.3E-01 0.828 1.3E-01 M09 0.917 4.2E-01
0.668 1.5E-02 0.728 3.5E-02 M10 0.980 6.6E-01 0.905 1.1E-01 0.923
1.9E-01 M11 0.995 9.4E-01 0.919 4.0E-01 0.924 3.5E-01 M12 0.964
5.1E-01 0.858 5.8E-02 0.890 1.6E-01 M13 0.948 3.5E-01 0.913 2.6E-01
0.963 6.3E-01 M14 0.864 1.1E-01 0.732 4.1E-02 0.847 1.4E-01 M15
0.947 2.6E-01 0.911 1.7E-01 0.962 5.2E-01 M16 0.957 5.5E-01 0.909
3.4E-01 0.950 5.8E-01 M17 0.877 1.4E-01 0.767 6.0E-02 0.875 2.6E-01
M18 0.932 4.5E-01 0.844 1.9E-01 0.905 3.9E-01 M19 0.891 2.8E-01
0.679 2.6E-02 0.762 4.5E-02 M20 0.896 8.6E-02 0.858 9.6E-02 0.957
5.9E-01 M21 0.937 4.2E-01 0.899 3.6E-01 0.959 6.8E-01 M22 0.926
3.4E-01 0.844 1.8E-01 0.912 4.0E-01 M23 0.931 3.9E-01 0.860 1.8E-01
0.923 4.5E-01 M24 0.840 1.2E-01 0.699 5.7E-02 0.832 1.5E-01 M25
0.949 6.7E-01 0.818 1.3E-01 0.862 4.4E-01
TABLE-US-00039 TABLE 39 Effect of Dementia State on Ethanolamine
Phospholipid Ratios to M01 in Females Cognitive Normal, SDAT_all,
SDAT, ADAS 5-19, SDAT, ADAS 20-39, SDAT, ADAS 40-70, Female Female
Female Female Female Metabolite Code Mean SEM Mean SEM Mean SEM
Mean SEM Mean SEM M01 1.000 0.000 1.000 0.000 1.000 0.000 1.000
0.000 1.000 0.000 M02 0.520 0.029 0.463 0.012 0.454 0.021 0.499
0.023 0.431 0.018 M03 0.852 0.034 0.687 0.017 0.742 0.030 0.704
0.031 0.624 0.022 M04 0.222 0.013 0.219 0.010 0.215 0.013 0.241
0.021 0.197 0.015 M05 0.092 0.004 0.086 0.003 0.091 0.004 0.088
0.005 0.081 0.004 M06 0.234 0.012 0.207 0.007 0.226 0.011 0.217
0.013 0.180 0.009 M07 0.474 0.034 0.368 0.018 0.422 0.039 0.377
0.028 0.314 0.025 M08 0.054 0.004 0.047 0.002 0.050 0.003 0.051
0.004 0.039 0.002 M09 0.167 0.013 0.121 0.005 0.140 0.010 0.123
0.009 0.103 0.008 M10 0.469 0.016 0.431 0.010 0.451 0.016 0.428
0.018 0.418 0.016 M11 0.929 0.039 0.807 0.023 0.886 0.041 0.839
0.044 0.708 0.030 M12 1.682 0.078 1.384 0.042 1.568 0.090 1.384
0.067 1.239 0.062 M13 0.111 0.005 0.095 0.003 0.101 0.005 0.095
0.005 0.089 0.004 M14 0.392 0.023 0.315 0.013 0.367 0.029 0.309
0.019 0.279 0.019 M15 0.627 0.024 0.539 0.014 0.564 0.021 0.563
0.030 0.493 0.016 M16 2.214 0.114 1.780 0.064 1.994 0.107 1.906
0.127 1.470 0.069 M17 3.497 0.247 2.593 0.127 2.976 0.266 2.750
0.226 2.115 0.146 M18 0.202 0.014 0.161 0.007 0.178 0.011 0.170
0.014 0.138 0.008 M19 0.895 0.061 0.634 0.030 0.728 0.057 0.656
0.055 0.537 0.041 M20 0.669 0.035 0.535 0.018 0.587 0.032 0.553
0.036 0.474 0.022 M21 2.318 0.118 1.798 0.076 2.039 0.132 1.922
0.147 1.467 0.086 M22 8.068 0.596 6.310 0.304 7.041 0.600 6.681
0.568 5.313 0.360 M23 0.158 0.009 0.127 0.005 0.139 0.009 0.131
0.010 0.112 0.006 M24 0.893 0.068 0.651 0.034 0.724 0.063 0.669
0.059 0.573 0.053 M25 2.145 0.127 2.031 0.086 2.072 0.129 2.109
0.162 1.910 0.139
TABLE-US-00040 TABLE 40 Ratio and T-test values of Ethanolamine
Phospholipid Ratios to M01 between females of various levels of
dementia AD, All to CN, Female ADAS 5-19 to CN, Female ADAS 20-39
to CN, Female ADAS 40-70 to CN, Female Metabolite Code Ratio ttest
Ratio ttest Ratio ttest Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0!
1.000 #DIV/0! 1.000 #DIV/0! M02 0.891 4.7E-02 0.872 6.4E-02 0.960
5.7E-01 0.828 7.0E-03 M03 0.806 1.3E-05 0.871 1.6E-02 0.826 2.1E-03
0.732 6.8E-08 M04 0.986 8.8E-01 0.968 7.0E-01 1.086 4.9E-01 0.887
2.3E-01 M05 0.941 3.5E-01 0.989 8.7E-01 0.962 6.4E-01 0.879 5.5E-02
M06 0.885 6.6E-02 0.966 6.3E-01 0.928 3.7E-01 0.771 2.8E-04 M07
0.776 6.7E-03 0.890 3.2E-01 0.796 3.2E-02 0.663 2.3E-04 M08 0.869
1.3E-01 0.934 4.9E-01 0.943 6.2E-01 0.735 1.5E-03 M09 0.724 3.1E-04
0.837 1.0E-01 0.740 5.4E-03 0.618 4.2E-05 M10 0.919 7.3E-02 0.963
4.4E-01 0.913 1.1E-01 0.892 3.1E-02 M11 0.868 1.6E-02 0.953 4.4E-01
0.902 1.5E-01 0.762 2.0E-05 M12 0.823 1.5E-03 0.932 3.4E-01 0.823
5.0E-03 0.736 2.0E-05 M13 0.854 6.6E-03 0.914 1.8E-01 0.854 3.1E-02
0.806 9.4E-04 M14 0.802 5.5E-03 0.935 4.9E-01 0.789 6.6E-03 0.711
2.4E-04 M15 0.860 4.8E-03 0.900 5.1E-02 0.897 1.3E-01 0.786 9.1E-06
M16 0.804 2.1E-03 0.901 1.6E-01 0.861 9.3E-02 0.664 8.8E-08 M17
0.742 1.5E-03 0.851 1.6E-01 0.786 3.2E-02 0.605 2.3E-06 M18 0.798
9.5E-03 0.879 1.8E-01 0.844 1.4E-01 0.683 6.3E-05 M19 0.709 1.6E-04
0.813 5.1E-02 0.733 5.3E-03 0.600 3.2E-06 M20 0.800 1.0E-03 0.878
8.7E-02 0.826 2.8E-02 0.709 3.4E-06 M21 0.776 1.4E-03 0.879 1.2E-01
0.829 5.6E-02 0.633 5.4E-08 M22 0.782 9.6E-03 0.873 2.3E-01 0.828
1.1E-01 0.659 7.8E-05 M23 0.801 5.3E-03 0.877 1.3E-01 0.831 6.8E-02
0.707 4.5E-05 M24 0.729 1.6E-03 0.811 7.2E-02 0.750 1.7E-02 0.642
3.3E-04 M25 0.947 5.3E-01 0.966 6.9E-01 0.983 8.7E-01 0.890
2.3E-01
TABLE-US-00041 TABLE 41 Ratio and T-test values of Ethanolamine
Phospholipid Ratios to M01 between females of various levels of
dementia ADAS 20-39 to 5-19, Female ADAS 40-70 to 5-19, Female ADAS
40-70 to 20-39, Female Metabolite Code Ratio ttest Ratio ttest
Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 1.101
1.6E-01 0.949 4.1E-01 0.862 2.2E-02 M03 0.949 4.0E-01 0.841 1.5E-03
0.887 4.2E-02 M04 1.122 3.3E-01 0.917 3.8E-01 0.817 9.2E-02 M05
0.972 7.3E-01 0.889 7.6E-02 0.914 2.6E-01 M06 0.961 6.3E-01 0.797
1.3E-03 0.830 2.4E-02 M07 0.895 3.4E-01 0.745 1.7E-02 0.833 9.8E-02
M08 1.010 9.4E-01 0.787 1.1E-02 0.780 3.5E-02 M09 0.884 2.4E-01
0.738 6.7E-03 0.835 1.0E-01 M10 0.948 3.5E-01 0.926 1.5E-01 0.977
6.8E-01 M11 0.947 4.6E-01 0.799 6.4E-04 0.844 1.9E-02 M12 0.883
9.7E-02 0.790 2.5E-03 0.895 1.2E-01 M13 0.934 3.6E-01 0.882 5.3E-02
0.944 4.1E-01 M14 0.844 8.6E-02 0.761 9.9E-03 0.902 2.6E-01 M15
0.998 9.7E-01 0.874 7.7E-03 0.876 5.3E-02 M16 0.956 6.2E-01 0.737
5.0E-05 0.771 4.2E-03 M17 0.924 5.2E-01 0.711 3.6E-03 0.769 2.4E-02
M18 0.960 7.2E-01 0.777 4.1E-03 0.809 5.7E-02 M19 0.901 3.8E-01
0.738 7.1E-03 0.819 9.2E-02 M20 0.941 4.9E-01 0.808 3.7E-03 0.858
7.3E-02 M21 0.943 5.8E-01 0.720 3.1E-04 0.763 1.1E-02 M22 0.949
6.7E-01 0.755 1.2E-02 0.795 5.1E-02 M23 0.948 6.1E-01 0.806 1.4E-02
0.851 1.1E-01 M24 0.925 5.4E-01 0.791 6.9E-02 0.856 2.3E-01 M25
1.018 8.7E-01 0.922 4.0E-01 0.906 3.6E-01
TABLE-US-00042 TABLE 42 Effect of Pathology State on Ethanolamine
Phospholipid Ratios to M01 in Males Post Mortem Ctl, Male Post
Mortem SDAT Male SDAT vs Control Metabolite Code Mean SEM Mean SEM
Ratio ttest M01 1.000 0.000 1.000 0.000 1.000 #DIV/0! M02 0.367
0.029 0.290 0.029 0.791 0.076 M03 0.482 0.042 0.391 0.013 0.811
0.054 M04 0.143 0.031 0.076 0.009 0.529 0.052 M05 0.048 0.009 0.029
0.002 0.607 0.052 M06 0.080 0.014 0.046 0.003 0.581 0.033 M07 0.107
0.025 0.059 0.005 0.549 0.074 M08 0.024 0.005 0.012 0.001 0.511
0.037 M09 0.052 0.018 0.018 0.002 0.341 0.074 M10 0.337 0.038 0.269
0.013 0.798 0.107 M11 0.452 0.066 0.272 0.017 0.602 0.016 M12 0.819
0.130 0.616 0.024 0.753 0.143 M13 0.079 0.009 0.060 0.005 0.759
0.085 M14 0.212 0.039 0.115 0.010 0.542 0.026 M15 0.375 0.035 0.344
0.025 0.918 0.483 M16 0.792 0.128 0.627 0.047 0.791 0.240 M17 0.849
0.191 0.561 0.040 0.660 0.156 M18 0.085 0.011 0.056 0.004 0.656
0.018 M19 0.273 0.051 0.164 0.015 0.601 0.056 M20 0.254 0.027 0.187
0.009 0.737 0.028 M21 0.613 0.109 0.371 0.021 0.605 0.042 M22 1.890
0.379 1.199 0.083 0.634 0.092 M23 0.074 0.008 0.055 0.004 0.750
0.061 M24 0.241 0.039 0.142 0.011 0.588 0.025 M25 2.066 0.225 2.384
0.388 1.154 0.487
TABLE-US-00043 TABLE 43 Effect of Pathology State on Ethanolamine
Phospholipid Ratios to M01 in Females Post Mortem Ctl, Female Post
Mortem SDAT Female Autopsy AD vs. Control, Female Metabolite Code
Mean SEM Mean SEM Ratio ttest M01 1.000 0.000 1.000 0.000 1.000
#DIV/0! M02 0.322 0.028 0.367 0.047 1.140 0.440 M03 0.400 0.022
0.361 0.023 0.902 0.237 M04 0.086 0.012 0.092 0.019 1.069 0.798 M05
0.040 0.003 0.032 0.005 0.809 0.225 M06 0.069 0.008 0.059 0.017
0.855 0.605 M07 0.102 0.016 0.077 0.013 0.752 0.242 M08 0.017 0.002
0.017 0.002 0.981 0.922 M09 0.033 0.005 0.024 0.003 0.725 0.133 M10
0.290 0.025 0.291 0.027 1.003 0.981 M11 0.384 0.044 0.364 0.057
0.950 0.797 M12 0.731 0.096 0.699 0.059 0.957 0.777 M13 0.061 0.008
0.069 0.007 1.122 0.489 M14 0.144 0.015 0.155 0.033 1.071 0.791 M15
0.339 0.021 0.301 0.027 0.889 0.297 M16 0.670 0.066 0.599 0.112
0.895 0.605 M17 0.697 0.063 0.591 0.082 0.848 0.329 M18 0.081 0.014
0.069 0.008 0.854 0.462 M19 0.211 0.017 0.180 0.018 0.853 0.230 M20
0.283 0.025 0.193 0.032 0.680 0.042 M21 0.624 0.079 0.421 0.127
0.674 0.203 M22 1.782 0.251 1.480 0.225 0.831 0.382 M23 0.073 0.011
0.060 0.006 0.816 0.275 M24 0.214 0.018 0.149 0.016 0.697 0.016 M25
1.382 0.214 1.767 0.221 1.279 0.229
TABLE-US-00044 TABLE 44 Effect of Dementia State on White and Gray
Matter Scores in Males White Matter Gray Matter Score Score Mean
SEM Mean SEM Cohort Cognitive Normal, Male -0.25 0.08 -0.44 0.11
SDAT_all, Male -0.63 0.06 -1.11 0.08 SDAT, ADAS 5-19, Male -0.56
0.09 -1.00 0.13 SDAT, ADAS 20-39, Male -0.67 0.08 -1.11 0.10 SDAT,
ADAS 40-70, Male -0.71 0.14 -1.45 0.16 Post Mortem Ctl, Male -0.44
0.13 -0.41 0.21 Post Mortem SDAT Male -1.62 0.19 -1.28 0.23
Comparison AD, All to CN, Male Delta -0.37 Delta -0.67 ttest
1.9E-03 ttest 2.4E-05 ADAS 5-19 to CN, Male Delta -0.31 Delta -0.56
ttest 1.8E-02 ttest 2.5E-03 ADAS 20-39 to CN, Male Delta -0.41
Delta -0.67 ttest 1.5E-03 ttest 7.8E-05 ADAS 40-70 to CN, Male
Delta -0.46 Delta -1.01 ttest 3.7E-03 ttest 1.8E-06 ADAS 20-39 to
5-19, Male Delta -0.10 Delta -0.11 ttest 4.2E-01 ttest 5.0E-01 ADAS
40-70 to 5-19, Male Delta -0.15 Delta -0.45 ttest 3.9E-01 ttest
5.2E-02 ADAS 40-70 to 20-39, Male Delta -0.04 Delta -0.34 ttest
8.0E-01 ttest 1.1E-01 Autopsy AD vs. Control, Male Delta -0.89
Delta -1.13 ttest 8.9E-03 ttest 2.8E-03
TABLE-US-00045 TABLE 45 Effect of Dementia State on White and Gray
Matter Scores in Females White Matter Gray Matter Score Score Mean
SEM Mean SEM Cohort Cognitive Normal, Female -0.27 0.09 -0.42 0.13
SDAT_all, Female -0.73 0.05 -1.01 0.07 SDAT, ADAS 5-19, Female
-0.55 0.08 -0.85 0.11 SDAT, ADAS 20-39, Female -0.69 0.09 -0.94
0.11 SDAT, ADAS 40-70, Female -0.91 0.08 -1.21 0.12 Post Mortem
Ctl, Female -0.50 0.29 -0.53 0.26 Post Mortem SDAT Female -1.34
0.27 -1.54 0.25 Comparison AD, All to CN, Female Delta -0.46 Delta
-0.59 ttest 4.6E-05 ttest 1.1E-04 ADAS 5-19 to CN, Female Delta
-0.29 Delta -0.43 ttest 1.7E-02 ttest 1.3E-02 ADAS 20-39 to CN,
Female Delta -0.43 Delta -0.52 ttest 1.9E-03 ttest 3.6E-03 ADAS
40-70 to CN, Female Delta -0.64 Delta -0.79 ttest 9.5E-07 ttest
2.2E-05 ADAS 20-39 to 5-19, Female Delta -0.14 Delta -0.09 ttest
2.7E-01 ttest 5.8E-01 ADAS 40-70 to 5-19, Female Delta -0.36 Delta
-0.37 ttest 2.9E-03 ttest 3.1E-02 ADAS 40-70 to 20-39, Female Delta
-0.21 Delta -0.27 ttest 9.4E-02 ttest 1.0E-01 Autopsy AD vs.
Control, Female Delta -1.17 Delta -0.81 ttest 2.0E-03 ttest
2.5E-02
TABLE-US-00046 TABLE 46 Distribution of White and Gray Matter
Scores in Males (Mean Normalized to CN Male) ADAS-cog ADAS-cog
ADAS-cog Autopsy Autopsy MMSE .gtoreq. 28 8-19 20-39 40-70 Control
AD Bin Frequency Frequency Frequency Frequency Frequency Frequency
White Matter Distribution -2 0 1 1 0 0 2 -1.75 0 0 0 0 0 0 -1.5 0 0
3 1 0 0 -1.25 0 2 4 0 0 1 -1 1 2 3 1 0 2 -0.75 1 3 3 4 0 2 -0.5 3 2
6 4 2 0 -0.25 3 8 13 4 3 1 0 7 13 12 0 2 1 0.25 9 6 5 1 2 1 0.5 5 1
5 2 0 0 0.75 1 0 1 0 1 0 1 2 0 2 1 0 0 More 0 2 0 0 0 0 Gray Matter
distribution -2 0 1 4 2 0 2 -1.75 0 2 2 1 0 0 -1.5 0 1 3 0 0 0
-1.25 1 4 2 2 0 1 -1 0 6 7 4 0 2 -0.75 1 4 9 2 1 3 -0.5 7 3 5 4 1 1
-0.25 3 5 9 1 2 0 0 3 2 7 1 1 0 0.25 5 4 2 1 3 0 0.5 7 3 4 0 1 1
0.75 3 1 2 0 0 0 1 0 3 2 0 0 0 More 2 1 0 0 1 0
TABLE-US-00047 TABLE 47 Distribution of White and Gray Matter
Scores in Females (Mean Normalized to CN Female) ADAS-cog ADAS-cog
ADAS-cog Autopsy Autopsy MMSE .gtoreq. 28 8-19 20-39 40-70 Control
AD Bin Frequency Frequency Frequency Frequency Frequency Frequency
White Matter Distribution -2 0 0 0 1 0 1 -1.75 0 0 0 0 0 1 -1.5 0 0
3 4 1 3 -1.25 1 0 2 1 0 0 -1 1 2 8 4 0 3 -0.75 1 5 6 11 1 0 -0.5 2
6 6 10 1 1 -0.25 4 8 6 3 2 1 0 9 5 8 9 1 0 0.25 8 6 5 3 2 0 0.5 3 3
6 1 0 0 0.75 4 3 1 1 0 0 1 3 0 2 0 0 0 More 0 0 1 0 1 0 Gray Matter
distribution -2 0 0 1 4 0 1 -1.75 0 1 2 1 0 1 -1.5 1 1 3 6 0 0
-1.25 2 2 4 6 0 0 -1 2 3 8 4 1 2 -0.75 1 5 6 5 0 1 -0.5 1 6 4 6 1 2
-0.25 5 6 7 3 2 0 0 4 5 4 3 2 2 0.25 6 3 6 6 2 1 0.5 5 2 2 1 0 0
0.75 3 1 2 2 0 0 1 3 2 2 0 0 0 More 3 1 3 1 1 0
TABLE-US-00048 TABLE 48 Effect of Age on White and Gray Matter
Scores in Males White Matter Gray Matter Score Score Mean SEM Mean
SEM Cohort Age Ctl, 30-39, Male -0.25 0.29 -0.92 0.28 Age Ctl,
40-49, Male -0.48 0.10 -1.28 0.14 Age Ctl, 50-59, Male -0.47 0.08
-0.90 0.11 Age Ctl, 60-69, Male -0.53 0.10 -0.84 0.14 Age Ctl,
70+_Male -0.43 0.09 -0.78 0.14 Comparison 50-59 vs. 40-49, Male
Delta 0.01 Delta 0.38 ttest 9.4E-01 ttest 4.2E-02 60-69 vs. 40-49,
Male Delta -0.05 Delta 0.45 ttest 7.4E-01 ttest 3.2E-02 70+ vs.
40-49, Male Delta 0.05 Delta 0.50 ttest 7.2E-01 ttest 1.4E-02
TABLE-US-00049 TABLE 49 Effect of Age on White and Gray Matter
Scores in Females White Matter Gray Matter Score Score Mean SEM
Mean SEM Cohort Age Ctl, 30-39, Female -0.36 0.18 -0.56 0.27 Age
Ctl, 40-49, Female -0.33 0.10 -0.61 0.13 Age Ctl, 50-59, Female
-0.55 0.06 -0.71 0.09 Age Ctl, 60-69, Female -0.62 0.09 -0.90 0.12
Age Ctl, 70+_Female -0.47 0.11 -0.58 0.14 Comparison 50-59 vs.
40-49, Female Delta -0.22 Delta -0.10 ttest 6.9E-02 ttest 5.4E-01
60-69 vs. 40-49, Female Delta -0.29 Delta -0.30 ttest 3.6E-02 ttest
1.1E-01 70+ vs. 40-49, Female Delta -0.14 Delta 0.03 ttest 3.7E-01
ttest 8.8E-01
TABLE-US-00050 TABLE 50 Risk prediction in Males Stats CN Low
Moderate Severe PM Ctl PM SDAT White Matter White Matter White
Matter White Matter White Matter White Matter Score Score Score
Score Score Score Control Control Control Control Control Control
Normalized Normalized Normalized Normalized Normalized Normalized
Total N 32 40 58 18 10 10 Total L 27 30 38 8 8 3 Total H 5 10 20 10
2 7 L % 84.4 75.0 65.5 44.4 80.0 30.0 Gray Matter Gray Matter Gray
Matter Gray Matter Gray Matter Gray Matter Score Score Score Score
Score Score Control Control Control Control Control Control
Normalized Normalized Normalized Normalized Normalized Normalized
Total N 32 40 58 18 10 10 Total L 23 19 26 3 8 1 Total H 9 21 32 15
2 9 L % 71.9 47.5 44.8 16.7 80.0 10.0 Low risk LL 22 19 25 3 7 1 %
LL 68.8 47.5 43.1 16.7 70.0 10.0 Intermediate risk IM 6 11 14 5 2 2
% IM 18.8 27.5 24.1 27.8 20.0 20.0 High risk HH 4 10 19 10 1 7 % HH
12.5 25.0 32.8 55.6 10.0 70.0
TABLE-US-00051 TABLE 51 Risk Prediction in Females Stats CN Low
Moderate Severe PM Ctl PM SDAT White Matter White Matter White
Matter White Matter White Matter White Matter Score Score Score
Score Score Score Control Control Control Control Control Control
Normalized Normalized Normalized Normalized Normalized Normalized
Total N 36 38 54 48 9 10 Total L 31 25 29 17 6 1 Total H 5 13 25 31
3 9 L % 86.1 65.8 53.7 35.4 66.7 10.0 Gray Matter Gray Matter Gray
Matter Gray Matter Gray Matter Gray Matter Score Score Score Score
Score Score Control Control Control Control Control Control
Normalized Normalized Normalized Normalized Normalized Normalized
Total N 36 38 54 48 9 10 Total L 29 20 26 16 7 3 Total H 7 18 28 32
2 7 L % 80.6 52.6 48.1 33.3 77.8 30.0 Low risk LL 29 17 24 11 5 0 %
LL 80.6 44.7 44.4 22.9 55.6 0.0 Intermediate risk IM 2 11 7 11 3 4
% IM 5.6 28.9 13.0 22.9 33.3 40.0 High risk HH 5 10 23 26 1 6 % HH
13.9 26.3 42.6 54.2 11.1 60.0
TABLE-US-00052 TABLE 52 Summary of key ratio and p-value statistics
for EtnPls 16:0/22:6 (M19) to PtdEt 16:0/18:0 (M01) serum ratio for
males and females combined. Comparison Ratio T-test 60-69 to 50-59
0.75 1.2E-02 70-95 to 50-59 0.95 6.4E-01 CN to 50-59 1.07 4.8E-01
SDAT to 50-59 0.70 4.7E-07 70-95 to 60-69 1.26 6.9E-02 CN to 60-69
1.42 3.8E-04 SDAT to 70-95 0.74 1.3E-04 SDAT to CN 0.65 7.6E-11
ADAS 5-19 to CN 0.74 3.0E-04 ADAS 20-39 to CN 0.66 1.3E-07 ADAS
40-70 to CN 0.53 3.9E-11 ADAS 20-39 to ADAS 5-19 0.90 1.6E-01 ADAS
40-70 to ADAS 5-19 0.72 3.4E-04 ADAS 40-70 to ADAS 20-39 0.80
1.0E-02 Post-Mortem SDAT to Control* 0.55 4.7E-03 *ratio and
p-value of EtnPls 16:0/22:6 alone.
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