Supplementary Table 1: Trabecular meshwork donors
Control Trabecular Meshwork donors
Code Age
Gender
Race**
SC001
SC002
34
31
M
M
W
W
Time
interval*
4h
5h
SC003
36
M
W
17h
SC004
70
F
W
9h
Myocardial infarction (MI)
SC005
SC006
SC007
62
67
63
M
F
M
W
W
W
5h
10h
16h
Cardiac arrest
Cardiac arrest
Respiratory failure
SC008
68
F
W
9h
COD, cancer
SC009
50
M
N/A
6h
CVA
SC010
57
M
W
7h
Thyroid cancer
SC011
SC012
SC013
60
68
67
M
M
M
N/A
B
W
3h
3h
10h
Cardiac arrhythmia
witnessed arrest
MI
SC014
SC015
SC016
SC017
59
54
64
55
M
M
M
M
W
W
W
W
16h
7h
11h
3h
Ischemic heart disease
Colon Cancer
MI
Vascular failure
SC018
SC019
35
74
F
F
W
W
8h
6h
Cardiopulmonary
CHF, respiratory failure
SC020
52
F
W
6h
MI, Heart disease
Cause of Death
Medical history
Cardiovascular Disease
No recorded history found
Self inflicted gun shot wound to the
PTSD, Ulcerative colitis, depression, colostomy
head
Cardiac Arrhythmia, Respiratory
hypertension, A Fib s/p ablation,
failure
hypersholesterolemia, sleep apnea, NIDDM, obesity
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MI, unremarkable history, vitamins, allergy
medication
Diabetes, Hypertension, arthritis
Cardiac arrest, no known history
Pulmonary fibrosis, Diabetes, CAD, Previous
incidence of MI
Advanced pulmonary emphysema-oxygen
dependent; Lung cancer, GERD, psoriasis,
hypothyroidism, hyperlipidemia, HTN,
cholecystectomy with incidental appendectomy,
osteoarthritis, tobacco use
Severe diabetes, hypercholesterolemia, below knee
amputation, spinal stenosis S/P surgery, arterial
peripheral bypass, free fibula graft, hip replacement,
S/P colostomy, prior seizures due to high blood
sugar.
Thyroid cancer, tracheostomy, PEG tube,
Thyroidectomy
Dyslipidemia, Diabetes, hypertension
Prostate cancer, COPD
CAD. CABG, HTN, Hypercholesterolemia, Prostate
CA, HIP FX, Illeostomy, Keratosis, Rhinitis
Hypertension, high cholesterol
Hypertension, Cancer
Appendectomy
HTN, Renal failure, hypothyroidism,
hypercholesterolemia, Chronic obstructive
pulmonary disease (COPD), S/P ankle fx repair
None recorded
MI, HTN, hypercholesterolemia, CHF, Uterine CA,
DM, CKD, right great toe amputation, hysterectomy
DM, HTN, Chronic lymphedema, bilateral cellulitis
on legs, emphysema, morbid obesity
SC021
SC022
SC023
41
56
67
F
M
M
W
12h
8h
15h
Cancer, Brain Glioblastoma
MI
Lung cancer
SC024
SC025
75
40
F
F
W
W
11h
11h
Trauma
Cardiac arrest, IHC/ Stroke
SC026
56
M
W
16h
MI
SC027
SC028
60
69
M
F
W
4h
7h
Anoxia, Cardiac arrhythmia
Cardiogenic shock, Acute MI
SC029
SC030
19
67
F
M
W
W
8h
8h
Cardiac arrest
Trauma
Code
SC031
SC032
SC033
SC034
Non-contributory
MI, CABPG, Sleep apnea, Shortness of breath
Cancer, coronary and periphral vascular disease,
CHF, COPD, hypertension, dyslipidemia
Brain injury, Celiac disease, NIDDM, Rosacea
Alagille's syndrome, heart disorder, pulmonary
stenosis, liver disease with jaundace secondary to
Alagille
s/p bypass surgery, Hypercholesterolemia, NIDDM,
COPD
Dyslipidemia, hypertension, NIDDM
NIDDM, Hypertension, Hyperlipidemia, CAD, MI with
4 vessels bypass, Osteopenia, CHF
Seizure disorder
NIDDM
Sphingomyelin
Control Trabecular Meshwork donors used for specific class of lipids
Sphingoid base
Sphingoid base-1-phosphate
Ceramides
SC006
SC010
SC015
SC016
SC017
SC018
SC019
SC020
SC001
SC051
SC21
SC22
SC23
SC24
SC25
SC26
SC27
SC28
SC29
SC30
SC010
SC014
SC015
SC016
SC017
SC018
SC019
SC020
SC001
SC002
SC21
SC22
SC23
SC24
SC25
SC26
SC27
SC28
SC29
SC30
Age
Gender
Race
81
72
76
84
F
M
M
M
W
W
B
W
SC003
SC004
SC005
SC007
SC008
SC051
SC009
SC011
SC012
SC013
SC21
SC22
SC23
SC24
SC25
SC26
SC27
SC28
SC29
SC30
Time
interval*
N/A
6.5
8
10
SC003
SC004
SC005
SC007
SC009
SC008
SC002
SC011
SC012
SC013
SC21
SC22
SC23
SC24
SC25
SC26
SC27
SC28
SC29
SC30
Glaucomatous Trabecular meshwork donors
Cause of Death
Not applicable, Surgical sample
Cardiac arrest
Unknown
Myocardial infraction (MI)
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Glaucoma medication†
MMC,beta blocker, PG, steroid, implant
Cosopt, PG
Cosopt, PG
brimonidine, Cosopt, PG, steriod, implant
SC035
SC036
SC037
SC038
SC039
SC040
SC041
SC042
SC043
SC044
SC045
SC046
SC047
SC048
SC049
SC050
SC051
SC052
71
78
81
85
81
75
59
67
59
67
55
50
57
51
80
73
75
62
F
M
F
F
M
F
F
F
M
M
M
M
F
F
M
F
F
F
W
W
B
W
W
W
W
B
W
W
W
W
W
B
W
W
W
W
9
N/A
6
6.5
7
8
4
8
6
6
6
6.5
5
12
16h
7.5h
5h
6h
SC053
SC054
SC055
SC056
SC057
SC058
62
73
67
75
73
77
F
F
M
F
F
F
W
W
W
W
W
W
6h
12h
10h
6h
8h
10h
Sphingomyelin
SC041
SC043
SC044
SC045
SC046
SC047
SC048
SC031
SC033
SC032
SC049
SC050
SC051
SC052
SC053
SC054
Cardiac arrest
Not applicable, Surgical sample
Myocardial infraction (MI)
Respiratory failure
MI
Multiple organ failure
MI
Cardiac arrest
Cardiac arrest
Unknown
Respiratory failure
MI
MI
Cardiac arrest
Multiple myeloma
Whole body inflammation
Liver and renal failure
Anoxic Encephalopathy
Cosopt
beta-blocker,brimonidine
beta-blocker,brimonidine, PG
brimonidine, Cosopt, PG
beta-blocker,brimonidine
beta-blocker, PG
MMC,beta blocker, brimonidine, PG,
Not known
beta blocker, brimonidine
MMC, Cosopt
Cosopt
MMC,beta-blocker, CAI, miotic, PG, steriod
brimonidine, PG
MMC,Cosopt, PG, implant, steroid
Xalatan
Timolol, Cosopt
Not known
Hypertension, hyperlipidemia, periphral vascular
disease, depression, insomnia, urinary incontinence,
arthropathy, osteoprosis, anemia
Anoxic Encephalopathy
No medication
Sepsis
Timolol
Metastatic Small Cell Lung Cancer
Timolol
Multi system organ failure
Not known
Sepsis
Timolol
Intracranial hemorrhage
Xalatan, timolol
Glaucomatous Trabecular Meshwork donors used for specific class of lipids
Sphingoid base
Sphingoid base-1-phosphate
Ceramides
SC041
SC043
SC044
SC046
SC035
SC047
SC048
SC034
SC031
SC032
SC049
SC050
SC051
SC052
SC053
SC054
SC032
SC033
SC034
SC035
SC036
SC037
SC038
SC039
SC040
SC042
SC049
SC050
SC051
SC052
SC053
SC054
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SC032
SC033
SC034
SC035
SC036
SC037
SC038
SC039
SC040
SC042
SC049
SC050
SC051
SC052
SC053
SC054
SC055
SC056
SC057
SC058
SC055
SC056
SC057
SC058
SC055
SC056
SC057
SC058
SC055
SC056
SC057
SC058
*Time interval is the time from death to enucleation. Eyes were preserved in phosphate buffered saline upon enucleation, maintained at 4ºC while transported. †The
glaucoma tissue donors did not suffer from diabetes or hyperlimidemia or were not statin users. Control and Glaucomatous tissue used for specific analyses of a particular
class of lipid (Sphingomyelin, Sphingoid base, Sphingoid base-1-phosphate, Ceramides ) has been indicated with a code and placed below the specific lipid class.
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Supplementary Table 2: Mass spectrometric parameters utilized
for confirmatory detection of Selected Fumonisin species
Species
Precursor ion
(m/z)[MH]+
Fumonisin B2
706
Fumonisin B3
706
Collision
energy (V)
40
38
29
33
Product ion
(m/z)
336
318
688
354
The Fumonisin species noted above was confirmed using precursor and
product ion detection at collision energies in a TSQ Quantum Access Max triple
quadrupole mass spectrometer in positive ion mode. Precursor ion was also
detected in presence of sodium or potassium ion, for example Fumonisin B1
[MNa]+ m/z is 744 and that of [MK]+ is 760.
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Supplementary Table 3: Common sphingomyelin species in the trabecular meshwork
Lipid Species*
C17 sphingosine-1-phosphocholine
Sphingosine-1-phosphocholine
Sphinganine-1-phosphocholine
SM(d18:1/12:0)
SM(d18:0/12:0)
SM(d18:0/13:0)
SM(d18:2/14:0)
SM(d16:1/16:0)
SM(d18:0/14:0)
SM(d18:2/15:0)
SM(d16:1/17:0)
SM(d18:0/15:0)
SM(d18:1/16:1)
SM(d18:1/16:0)
SM(d18:0/16:0)
SM(d18:1/17:0)
SM(d18:0/17:0)
SM(d18:2/18:1)
SM(d18:1/18:1(9Z))
SM(d18:1/18:0)
SM(d16:0/20:0)
SM(d18:1/19:0)
SM(d18:2/20:1)
SM(d18:1/20:1)
SM(d16:1/22:0)
SM(d18:0/20:0)
SM(d18:2/21:0)
SM(d17:1/22:0)
SM(d18:1/22:1)
SM(d18:1/22:0)
SM(d18:0/22:0)
SM(d17:1/24:1)
SM(d16:1/25:0)
SM(d18:2/24:1)
Number of
Isomers
N/A
N/A
N/A
2
N/A
N/A
3
4
N/A
3
3
N/A
5
5
N/A
3
N/A
N/A
7
5
2
3
N/A
2
1
1
N/A
4
2
1
N/A
1
2
N/A
Exact
m/z
450.3
464.3
466.3
646.5
648.5
662.5
672.5
674.5
676.5
686.5
688.5
690.5
700.5
702.5
704.5
716.5
718.5
726.5
728.5
730.6
732.6
744.6
754.6
756.6
758.6
760.6
770.6
772.6
784.6
786.6
788.6
798.6
800.6
810.6
m/z** Actual
[M+H]+
m/z
451.3
450.9
465.3
465.8
467.3
467.1
647.5
647.9
649.5
649.1
663.5
663.2
673.5
673.2
675.5
675.6
677.5
677.4
687.5
687.9
689.5
689.1
691.5
691.8
701.5
701.2
703.5
703.7
705.5
705.6
717.5
716.9
719.6
719.3
727.5
726.8
729.5
730.1
731.6
731.3
733.6
733.7
745.6
746.6
755.6
754.8
757.6
757.9
759.6
759.3
761.6
761.1
771.6
771.8
773.6
773.2
785.6
786.2
787.6
788.1
789.6
789.1
799.6
799.7
801.6
801.2
811.6
811.9
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Error
(Da)
0.4
-0.5
0.2
-0.4
0.4
0.3
0.3
-0.1
0.1
-0.4
0.4
-0.3
0.3
-0.2
-0.1
0.6
0.3
0.7
-0.6
0.3
-0.1
-1.0
0.8
-0.3
0.3
0.5
-0.2
0.4
-0.6
-0.5
0.5
-0.1
0.4
-0.3
Control
Average
SEM
Donor
(pmol/ug)
Frequency
2.30
0.53
6
5.42
1.24
6
18.05
4.14
7
54.50
12.50
13
94.99
21.79
10
10.85
2.49
15
26.01
5.97
9
22.08
5.07
14
13.42
3.08
9
3.60
0.83
15
8.27
1.90
8
21.90
5.02
3
28.75
6.60
8
38.08
8.74
13
41.58
9.54
10
10.45
2.40
16
20.19
4.63
9
11.41
2.62
11
38.53
8.84
4
0.11
0.03
2
7.86
1.80
6
4.18
0.96
16
4.98
1.14
9
14.14
3.24
7
22.67
5.20
17
14.78
3.39
9
45.75
10.50
2
14.41
3.31
3
3.37
0.77
5
5.85
1.34
15
8.16
1.87
8
218.28
50.08
7
2.73
0.63
7
5.56
1.28
8
Glaucoma
Average
SEM
Donor
(pmol/ug)
Frequency
4.62
1.03
6
45.67
10.21
6
3.63†
0.81
6
3.1†
0.69
13
64.26
14.37
7
2.03†
0.45
12
2.07†
0.46
8
18.30
4.09
12
12.71
2.84
7
3.69
0.83
13
1.67
0.37
8
19.11
4.27
4
86.42
19.32
8
7.95†
1.78
12
4.13†
0.92
10
3.35†
0.75
12
2.93†
0.66
10
12.30
2.75
11
3.36†
0.75
3
4.43†
0.99
1
19.63
4.39
5
10.55
2.36
13
10.07
2.25
10
3.98†
0.89
7
9.70
2.17
16
9.33
2.09
10
16.18
3.62
2
2.73†
0.61
5
9.30
2.08
5
12.03
2.69
14
3.36
0.75
9
14.34†
3.21
6
9.17
2.05
5
4.86
1.09
7
SM(d18:1/24:1(15Z))
SM(d18:0/24:1(15Z))
SM(d18:0/24:0)
SM(d19:1/24:1)
SM(d19:0/24:1)
SM(d18:1/26:1(17Z))
SM(d20:0/24:1)
SM(d17:0/27:0)
1
1
N/A
2
3
N/A
2
1
812.6
814.7
816.7
826.7
828.7
840.7
842.7
844.7
813.6
815.7
817.7
827.7
829.7
841.7
843.7
845.7
813.1
815.5
817.4
827.1
829.8
841.9
843.5
845.2
0.5
0.2
0.3
0.6
-0.1
-0.2
0.2
0.5
2.73
6.99
11.40
22.75
8.75
5.88
14.91
5.56
0.63
1.60
2.62
5.22
2.01
1.35
3.42
1.28
5
7
6
7
7
7
7
4
3.38
10.35
26.29
23.85
6.49
9.41
4.23
7.13
0.76
2.31
5.88
5.33
1.45
2.10
0.95
1.59
7
7
5
6
5
5
6
5
*The lipid species identification is based on Lipidmaps database, used as a *.csv file for bioinformatic analyses with MZmine 2.9 program. ** A representative
mass/charge ratio is presented (variations in m/z was reconciled by MZmine 2.9). For some lipid species identified, standard error of mean (SEM) could not be
calculated due to lack of presence in all samples. Average lipid amount is pmol lipid per species/µg protein. **The theoretical mass by charge ratio (m/z) presented
from Lipidmaps is in positive ion mode [M+H]+ for Sphingomyelin and sphingoid base. Whereas negative ion mode [M-H]- has been presented for Sphingoid base-1
Phosphate and Ceramide. Error in Da has been provided from theoretical data. Average standard normalized dataset is presented here. The selected common lipid
species had statistically significant differences between the control and POAG groups by ANOVA. The Scheffe’s post hoc test showed that they were statistically
different (†P ≤ 0.05).
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Supplementary Table 4: Common sphingoid base species in the trabecular meshwork
Lipid Species*
Xestoaminol C
(4E,6E,d14:2) sphingosine
C16 Sphinganine
Obscuraminol A
Prosafrinine
iso (4E,15-methyl-d16:1) sphingosine
C17 Sphinganine
(4E,8E,10E-d18:3)sphingosine
(4E,8Z,d18:2) sphingosine
Sphinganine
Clavepictine B
R-Dysidazirine
6-hydroxysphingosine
Phytosphingosine
N,N-dimethylsphingosine
Penaresidin A
N,N,N-trimethyl-sphingosine
Capnine
Penazetidine A
Sphingofungin E
Sphingofungin A
Sulfamisterin
Calyxinin
Hemsleyin imine A
Rhizochalinin D
Fumonisin B4
Fumonisin C3
Fumonisin B2
Fumonisin C1
Termitomycesphin A
Plakoside A
Number of
Isomers
N/A
N/A
N/A
1
N/A
3
N/A
N/A
6
2
N/A
N/A
2
N/A
1
1
N/A
N/A
N/A
N/A
N/A
N/A
1
1
N/A
N/A
1
1
N/A
N/A
N/A
Exact
m/z
229.2
241.2
273.2
277.2
283.2
285.2
287.2
295.2
297.2
301.3
305.2
307.2
315.2
317.3
327.3
329.3
342.3
351.2
369.3
417.2
431.3
461.2
486.4
605.5
646.5
689.4
691.3
705.4
707.3
743.5
947.7
m/z**
[M+H]+
230.2
242.2
274.2
278.2
284.2
286.2
288.2
296.2
298.2
302.3
306.2
308.2
316.2
318.3
328.3
330.3
343.3
352.2
370.3
418.2
432.3
462.2
487.4
606.5
647.5
690.4
692.3
706.4
708.3
744.5
948.7
Actual
m/z
230.5
242.6
274.1
278.9
284.3
285.8
288.5
296.4
298.4
302.7
306.1
307.7
315.2
318.2
328.7
330.9
344.1
352.9
369.5
418.5
432.1
462.4
487.1
606.2
647.8
690.5
692.1
706.5
708.7
744.4
948.5
Error
(Da)
-0.3
-0.4
0.1
-0.7
-0.1
0.4
-0.3
-0.2
-0.2
-0.4
0.1
0.5
1.0
0.1
-0.4
-0.6
-0.8
-0.7
0.8
-0.3
0.2
-0.2
0.3
0.3
-0.3
-0.1
0.2
-0.1
-0.4
0.1
0.2
Average
(pmol/ug)
11.78
140.25
83.02
10.62
0.80
68.26
68.93
23.84
0.57
201.51
3.85
22.29
9.66
32.99
1003.85
16.14
31.32
506.19
1056.27
682.80
202.21
460.08
84.83
302.27
121.95
167.28
33.49
127.42
2.82
43.53
0.57
Control
SEM
2.70
32.18
19.05
2.44
0.18
15.66
15.81
5.47
0.13
46.23
0.88
5.11
2.22
7.57
230.30
3.70
7.19
116.13
242.32
156.65
46.39
105.55
19.46
69.35
27.98
38.38
7.68
29.23
0.65
9.99
0.13
Donor
Frequency
14
16
14
9
7
10
16
5
6
2
5
13
10
16
16
4
13
19
15
17
16
17
15
18
15
18
11
17
7
13
6
Average
(pmol/ug)
8.32
7.55†
8.75†
8.99
2.18
0.59†
23.98
6.13†
8.55†
0.93†
6.30
91.50
719.47†
21.12
219.50
19.86
21.62
12.63†
160.88
21.60
35.56
157.70
32.19
17.09†
18.20†
10.84†
7.93†
4.09†
27.34†
33.42
4.84
Glaucoma
SEM
Donor
Frequency
1.86
15
1.69
16
1.96
15
2.01
7
0.49
4
0.13
9
5.36
14
1.37
5
1.91
7
0.21
4
1.41
8
20.46
12
160.88
9
4.72
16
49.08
12
4.44
6
4.83
15
2.82
15
35.97
14
4.83
15
7.95
17
35.26
15
7.20
15
3.82
12
4.07
15
2.42
16
1.77
10
0.91
13
6.11
10
7.47
16
1.08
5
*The lipid species identification is based on Lipidmaps database, used as a *.csv file for bioinformatic analyses with MZmine 2.9 program. ** A representative
mass/charge ratio is presented (variations in m/z was reconciled by MZmine 2.9). For some lipid species identified, standard error of mean (SEM) could not be
calculated due to lack of presence in all samples. Average lipid amount is pmol lipid per species/µg protein. **The theoretical mass by charge ratio (m/z) presented
from Lipidmaps is in positive ion mode [M+H]+ for Sphingomyelin and sphingoid base. Whereas negative ion mode [M-H]- has been presented for Sphingoid base-1
Phosphate and Ceramide. Error in Da has been provided from theoretical data. Average standard normalized dataset is presented here. The selected common lipid
species had statistically significant differences between the control and POAG groups by ANOVA. The Scheffe’s post hoc test showed that they were statistically
different (†P ≤ 0.05).
Downloaded From: http://iovs.arvojournals.org/pdfaccess.ashx?url=/data/journals/iovs/932990/ on 06/18/2017
Supplementary Table 5: Common Sphingoid base-1-phosphate species in the trabecular meshwork
Lipid Species*
C16 Sphingosine-1-phosphate
C16 Sphinganine-1-phosphate
C17 Sphingosine-1-phosphate
C17 Sphinganine-1-phosphate
Sphinganine-phosphate
C19 Sphingosine-1-phosphate
Phytosphingosine 1-phosphate
Number of
Isomers
Exact
m/z
m/z**
[M-H]-
Actual
m/z
Error
(Da)
N/A
N/A
N/A
N/A
N/A
N/A
N/A
351.2
353.2
365.2
367.2
381.2
393.2
397.2
350.2
352.2
364.2
366.2
380.2
392.2
396.2
350.5
352.8
364.1
366.6
379.2
392.8
396.1
-0.3
-0.6
0.1
-0.4
1.0
-0.6
0.1
Control
Average SEM
Donor
(pmol/ug)
Frequency
1.29
15.49
3.48
17.50
0.79
8.10
11.09
0.30
3.55
0.80
4.01
0.18
1.86
2.54
9
6
14
7
3
13
11
Glaucoma
Average
SEM
Donor
(pmol/ug)
Frequency
11.90†
5.26
11.45†
2.63†
3.46†
4.13
7.08
2.66
1.18
2.56
0.59
0.77
0.92
1.58
9
6
8
4
4
6
7
*The lipid species identification is based on Lipidmaps database, used as a *.csv file for bioinformatic analyses with MZmine 2.9 program. ** A representative
mass/charge ratio is presented (variations in m/z was reconciled by MZmine 2.9). For some lipid species identified, standard error of mean (SEM) could not be
calculated due to lack of presence in all samples. Average lipid amount is pmol lipid per species/µg protein. **The theoretical mass by charge ratio (m/z) presented
from Lipidmaps is in positive ion mode [M+H]+ for Sphingomyelin and sphingoid base. Whereas negative ion mode [M-H]- has been presented for Sphingoid base-1
Phosphate and Ceramide. Error in Da has been provided from theoretical data. Average standard normalized dataset is presented here. The selected common lipid
species had statistically significant differences between the control and POAG groups by ANOVA. The Scheffe’s post hoc test showed that they were statistically
different (†P ≤ 0.05).
Downloaded From: http://iovs.arvojournals.org/pdfaccess.ashx?url=/data/journals/iovs/932990/ on 06/18/2017
Supplementary Table 6: Common ceramides species in the trabecular meshwork
Lipid Species*
Number of
Isomers
Cer(d18:0/13:0)
N/A
Cer(d18:2/14:0)
2
Cer(d18:0/14:0)
N/A
Cer(d16:1/17:0)
N/A
Cer(d18:2/16:0)
3
Cer(d18:0/16:0)
N/A
Cer(d18:0/17:0)
N/A
CerP(d18:1/12:0)
N/A
Cer(d18:0/18:1(9Z))
3
Cer(d18:0/h17:0)
N/A
CerP(d18:1/14:0)
N/A
Cer(d18:2/20:0)
4
Cer(d16:1/22:0)
2
Cer(d18:2/21:0)
1
CerP(d18:0/16:0)
N/A
Cer(d18:1/22:0)
2
Cer(d18:2/23:0)
N/A
SM(d18:1/12:0)
2
SM(d18:0/12:0)
N/A
Cer(d18:0/24:1(15Z))
1
SM(d18:0/13:0)
N/A
Cer(d18:0/h24:0)
4
SM(d18:2/14:0)
3
SM(d18:1/14:0)
4
Cer(d18:1/26:1(17Z))
N/A
Cer(d18:1/26:0)
1
Cer(d18:0/26:0)
1
SM(d18:2/15:0)
3
SM(d18:1/15:0)
3
SM(d18:0/15:0)
N/A
Cer(t18:0/26:0)
4
SM(d18:1/16:1)
5
CerP(d18:1/22:0)
N/A
SM(d16:1/18:0)
5
SM(d18:0/16:0)
N/A
SM(d18:1/17:0)
3
SM(d18:0/17:0)
N/A
CerP(d18:1/24:1(15Z))
N/A
SM(d16:1/20:1)
7
CerP(d18:1/24:0)
N/A
SM(d18:0/18:1(9Z))
5
SM(d18:0/18:0)
2
SM(d19:1/18:0)
3
SM(d16:1/22:1)
2
SM(d18:1/20:0)
1
SM(d18:0/20:0)
1
SM(d18:2/21:0)
N/A
SM(d17:1/22:0)
4
SM(d18:2/22:1)
N/A
SM(d16:1/24:1)
2
SM(d16:1/24:0)
1
SM(d17:1/24:1)
1
SM(d16:1/25:0)
2
SM(d18:1/24:1(15Z))
1
Exact m/z** Actual
m/z [M-H]- m/z
497.4 496.4 496.2
507.4 506.4 506.9
511.4 510.4 510.1
523.4 522.4 522.3
535.4 534.4 534.6
539.5 538.5 538.8
553.5 552.5 552.3
561.4 560.4 560.5
565.5 564.5 563.5
569.5 568.5 568.9
589.4 588.4 589.1
591.5 590.5 590.1
593.5 592.5 592.2
605.5 604.5 604.8
619.4 618.4 618.5
621.6 620.6 620.3
633.6 632.6 632.4
646.5 645.5 645.7
648.5 647.5 647.9
649.6 648.6 647.7
662.5 661.5 661.8
667.6 666.6 666.1
672.5 671.5 671.2
674.5 673.5 673.8
675.6 674.6 674.3
677.6 676.6 676.1
679.6 678.6 678.7
686.5 685.5 685.2
688.5 687.5 687.1
690.5 689.5 689.1
695.6 694.6 695.6
700.5 699.5 699.2
701.5 700.5 700.8
702.5 701.5 701.1
704.5 703.5 703.8
716.5 715.5 715.1
718.5 717.5 717.9
727.5 726.5 726.3
728.5 727.5 727.9
729.6 728.5 728.4
730.5 729.5 729.8
732.6 731.6 731.4
744.6 743.6 743.2
756.6 755.6 754.6
758.6 757.6 757.5
760.6 759.6 759.8
770.6 769.6 769.9
772.6 771.6 771.3
782.6 781.6 781.2
784.6 783.6 783.7
786.6 785.6 785.1
798.6 797.6 797.2
800.6 799.6 799.8
812.6 811.6 811.1
Control
Error Average SEM
Donor
(Da) (pmol/ug)
Frequency
0.2
0.88
0.40
18
-0.5
5.48
3.55
20
0.3
115.13
51.03
11
0.1
5.07
2.61
19
-0.2
334.77 201.85
20
-0.3
433.51 114.92
9
0.2
2.19
0.74
18
-0.1
2.52
0.82
2
1.0
2.51
1.10
5
-0.4
1.97
0.69
9
-0.7
3.91
2.08
16
0.4
3.23
1.76
6
0.3
3.01
0.81
5
-0.3
0.65
0.25
15
-0.1
2.37
1.26
10
0.3
2.46
0.95
8
0.2
1.36
0.58
15
-0.2
1.78
0.83
15
-0.4
7.83
2.07
6
0.9
0.02
0.01
2
-0.3
2.54
1.68
16
0.5
0.58
0.18
15
0.3
0.89
0.37
10
-0.3
0.22
0.08
8
0.3
0.11
0.02
4
0.5
1.96
0.50
7
-0.1
0.64
0.24
6
0.3
1.46
0.36
10
0.4
2.00
0.91
6
0.4
0.06
0.01
6
-1.0
0.77
0.30
11
0.3
1.29
0.51
12
-0.3
0.63
0.21
5
0.4
0.47
0.21
6
-0.3
1.46
0.49
9
0.4
18.48
7.32
15
-0.4
0.04
0.00
1
0.2
0.97
0.34
16
-0.4
3.19
1.02
5
0.1
5.42
2.47
4
-0.3
3.48
0.66
2
0.2
0.35
0.11
6
0.4
0.60
0.17
12
1.0
0.50
0.17
15
0.1
1.11
0.28
5
-0.2
0.29
0.13
5
-0.3
1.23
0.45
13
0.3
1.05
0.21
6
0.4
0.68
0.28
13
-0.1
1.09
0.23
7
0.5
0.08
0.02
6
0.4
0.50
0.23
12
-0.2
0.79
0.36
7
0.5
1.56
0.71
14
Downloaded From: http://iovs.arvojournals.org/pdfaccess.ashx?url=/data/journals/iovs/932990/ on 06/18/2017
Glaucoma
Average SEM
Donor
(pmol/ug)
Frequency
1.13†
0.58
13
0.66
0.24
17
15.01† 5.07
12
0.97
0.34
13
5.93†
2.14
19
45.45† 11.89
10
1.24
0.62
17
2.11
0.64
7
1.62
0.38
4
0.57
0.16
12
0.97
0.38
15
0.51
0.16
8
2.22
0.56
6
0.35
0.16
14
0.38
0.10
14
0.55
0.14
6
0.70
0.23
18
0.45
0.13
16
0.32
0.02
5
2.50†
0.00
1
0.17
0.04
14
0.28
0.10
11
0.30
0.08
10
0.22
0.06
6
0.26
0.08
5
0.90
0.19
9
0.43
0.12
6
27.92† 22.24
13
0.29
0.13
6
0.21
0.06
7
1.55
0.94
11
0.67
0.39
11
0.09
0.02
3
0.003
0.00
1
0.17
0.05
6
0.76
0.17
16
0.43
0.13
9
0.99
0.37
14
0.01
0.00
2
0.25
0.07
3
0.11
0.02
3
0.55
0.17
5
0.53
0.27
14
0.34
0.12
16
0.84
0.49
8
0.08
0.02
8
0.68
0.42
14
0.19
0.06
8
0.60
0.18
15
0.46
0.24
7
0.40
0.14
3
0.15
0.03
12
0.05
0.01
5
0.30
0.15
13
SM(d18:1/24:0)
SM(d18:0/24:0)
SM(d18:2/25:0)
SM(d18:1/25:0)
SM(d18:1/26:1(17Z))
SM(d18:0/26:1(17Z))
SM(d18:0/26:0)
Ins-1-P-Cer(d18:1/22:0
1
N/A
2
3
N/A
2
1
N/A
814.6
816.7
826.6
828.7
840.7
842.7
844.7
863.6
813.6
815.7
825.6
827.7
839.7
841.7
843.7
862.6
813.2
815.5
825.5
827.8
839.9
841.5
844.3
863.5
0.4
0.2
0.1
-0.1
-0.2
0.2
-0.6
-0.9
9.90
7.73
0.76
1.04
0.81
0.37
0.28
0.52
5.67
4.84
0.44
0.44
0.45
0.11
0.10
0.24
9
10
15
7
10
7
8
11
0.23
4.30
0.35
0.12
0.48
0.31
0.08
0.41
0.10
1.31
0.11
0.03
0.15
0.06
0.02
0.19
11
7
13
6
11
8
5
12
*The lipid species identification is based on Lipidmaps database, used as a *.csv file for bioinformatic analyses with MZmine 2.9 program. ** A
representative mass/charge ratio is presented (variations in m/z was reconciled by MZmine 2.9). For some lipid species identified, standard
error of mean (SEM) could not be calculated due to lack of presence in all samples. Average lipid amount is pmol lipid per species/µg protein.
**The theoretical mass by charge ratio (m/z) presented from Lipidmaps is in positive ion mode [M+H]+ for Sphingomyelin and sphingoid base.
Whereas negative ion mode [M-H]- has been presented for Sphingoid base-1 Phosphate and Ceramide. Error in Da has been provided from
theoretical data. Average standard normalized dataset is presented here. The selected common lipid species had statistically significant
differences between the control and POAG groups by ANOVA. The Scheffe’s post hoc test showed that they were statistically different ( †P ≤
0.05).
Downloaded From: http://iovs.arvojournals.org/pdfaccess.ashx?url=/data/journals/iovs/932990/ on 06/18/2017
Control POAG
SB
Cer
SB1P
Control POAG
C17 sphingo sine-1-pho spho cho line
Cer(d16:1/17:0)
SM (d16:0/20:0)
(4E,8E,10E-d18:3)sphingo sine
Cer(d16:1/22:0)
Cer(d18:0/13:0)
SM (d16:0/22:0)
(4E,8Z,d18:2) sphingo sine
Cer(d18:0/14:0)
SM (d16:1/16:0)
6-hydro xysphingo sine
Cer(d18:0/16:0)
SM (d16:1/17:0)
Cer(d18:0/17:0)
C16 Sphinganine
Cer(d18:0/18:1(9Z))
SM (d16:1/18:1)
C17 Sphinganine
Cer(d18:0/24:1(15Z))
SM (d16:1/22:0)
Calyxinin
Cer(d18:0/26:0)
SM (d16:1/25:0)
Cer(d18:0/h17:0)
Capnine
Cer(d18:0/h24:0)
SM (d17:0/27:0)
Clavepictine B
Cer(d18:1/22:0)
SM (d17:1/17:0)
Fumo nisin B 2
Cer(d18:1/26:0)
SM (d17:1/22:0)
Cer(d18:1/26:1(17Z))
Fumo nisin B 4
SM (d17:1/24:1)
Cer(d18:2/14:0)
Fumo nisin C1
Cer(d18:2/16:0)
SM (d18:0/12:0)
Cer(d18:2/20:0)
Fumo nisin C3
SM (d18:0/13:0)
Cer(d18:2/21:0)
Hemsleyin imine A
Cer(d18:2/23:0)
SM (d18:0/14:0)
iso (4E,15-methyl-d16:1) sphingo sine
Cer(t18:0/26:0)
SM (d18:0/15:0)
CerP(d18:0/16:0)
N,N,N-trimethyl-sphingo sine
SM (d18:0/16:0)
CerP(d18:1/12:0)
N,N-dimethylsphingo sine
CerP(d18:1/14:0)
SM (d18:0/17:0)
Obscuramino l A
CerP(d18:1/22:0)
SM (d18:0/20:0)
CerP(d18:1/24:0)
P enaresidin A
SM (d18:0/22:0)
CerP(d18:1/24:1(15Z))
P enazetidine A
SM (d18:0/24:0)
Ins-1-P-Cer(d18:1/22:0)
SM(d16:1/18:0)
P hyto sphingo sine
SM (d18:0/24:1(15Z))
SM(d16:1/20:1)
P lako side A
SM (d18:1/12:0)
SM(d16:1/22:1)
P ro safrinine
SM (d18:1/14:0)
SM(d16:1/24:0)
SM(d16:1/24:1)
R-Dysidazirine
SM (d18:1/16:0)
SM(d16:1/25:0)
Rhizo chalinin D
SM (d18:1/16:1)
SM(d17:1/22:0)
Sphinganine
SM (d18:1/17:0)
SM(d17:1/24:1)
SM(d18:0/12:0)
SM (d18:1/18:1(9Z))
Sphingo fungin A
SM(d18:0/13:0)
SM (d18:1/19:0)
Sphingo fungin E
SM(d18:0/15:0)
SM
(d18:1/20:0)
SM(d18:0/16:0)
Sulfamisterin
SM(d18:0/17:0)
SM (d18:1/20:1)
Termito mycesphin A
SM(d18:0/18:0)
SM (d18:1/22:0)
Xesto amino l C
SM(d18:0/18:1(9Z))
SM (d18:1/22:1)
SM(d18:0/20:0)
SM(d18:0/24:0)
SM (d18:1/24:1(15Z))
SM(d18:0/26:0)
SM (d18:1/26:1(17Z))
SM(d18:0/26:1(17Z))
SM (d18:2/14:0)
SM(d18:1/12:0)
SM(d18:1/14:0)
Control POAG
SM (d18:2/15:0)
SM(d18:1/15:0)
C16 Sphinganine-1-pho sphate
SM (d18:2/18:1)
SM(d18:1/16:1)
C16 Sphingo sine-1-pho sphate
SM (d18:2/20:1)
SM(d18:1/17:0)
SM(d18:1/20:0)
C17 Sphinganine-1-pho sphate
SM (d18:2/21:0)
SM(d18:1/24:0)
C17 Sphingo sine-1-pho sphate
SM (d18:2/24:1)
SM(d18:1/24:1(15Z))
C19 Sphingo sine-1-pho sphate
SM (d19:0/24:1)
SM(d18:1/25:0)
SM(d18:1/26:1(17Z))
SM (d19:1/24:1)
P hyto sphingo sine 1-pho sphate
SM(d18:2/14:0)
SM (d20:0/24:1)
Sphinganine-pho sphate
SM(d18:2/15:0)
Sphinganine-1-pho spho cho line
SM(d18:2/21:0)
SM(d18:2/22:1)
Sphingo sine-1-pho spho cho line
SM(d18:2/25:0)
(4E,6E,d14:2) sphingo sine
.003 to 50
51 to 100
101 to 150
151 to 200
>200
Supplementary Figure 1. Heat map of common sphingolipids and ceramides species between control and POAG TM. Average
amounts pmol per normalized protein amount (µg) have been presented. A uniform scale bar for all classes (inset), as indicated have
been used. Abbreviations: Sphingomyelin (SM), sphingoid base (SB), sphingoid base-1-phosphate (SB1P) and ceramides (Cer).
Downloaded From: http://iovs.arvojournals.org/pdfaccess.ashx?url=/data/journals/iovs/932990/ on 06/18/2017
Control POAG
SM
TACTGANNGAGGGCCCTCTGGGTCNNGCTCCAACGCGAGTTTATTGGACCTTGTTGCNTTGGGGGGCCCGCCTTTA
CTGGTCGCCGGGGGGCTCACGCCCCCGGGNNCGCGCCCGGCNAAGACACCCTCGAACTCTGTCTGAAGATTGAA
GTCTGAGTGAAAATATAAATTATTTAAAACTTTCAACAACGGATCTCTTGGANNTATACACAACTTTCAACAACGGAT
CTCTTGGAACAACGGATCTCTTGGAACGGATCTCTTGGANNNACNGATCTTTTGGANTGCCTTGTCNCTCTCCNTNN
CTTCTTTTCTTTNCAGACTAGATATTTCCTAAGAATCTTGNCTGNNNCGTATGCCNTGACACACTGACCGACCTCATT
TTTGAATAAANTTGNTGCCAGTTNNTTTCCAGCATTTCNATACAGNNTCCTTTCGAGNGTTAATTNCCCGCTCCTGAG
CGNNTTNTACNNCCGANNCACGTTAGTTTNNTANCGCGGAGGCCAAGCCNCAAGCACCCCGAGAGATATGGCGCG
GGC
Query 1 CTCNTCANCCTGTGACATACCTAAAACGTTGCTTCGGCGGGAACAGACGGCCCCGTAACA 60
Subject 19 CTCATCACCCTGTGACATACCTAAAACGTTGCTTCGGCGGGAACAGACGGCCCCGTAACA 78
Query 61 CGGGCCGCCCCCGCCAGAGGACCCCCTAACTCTGTTTCTATTATGTTTCTTCTGAGTAAA 120
Subject 79 CGGGCCGCCCCCGCCAGAGGACCCCCTAACTCTGTTTCTATTATGTTTCTTCTGAGTAAA 138
Query 121 ACAAGCAAATAAATTAAAACTTTCAACAACGGATCTCTTGGCTCTGGCATCGATGAAGAA 180
Subject 139 ACAAGCAAATAAATTAAAACTTTCAACAACGGATCTCTTGGCTCTGGCATCGATGAAGAA 198
Query 181 CGCAGC 186
Subject 199 CGCAGC 204
Supplementary Figure 2. A representative sequence of a PCR product obtained using Fusarium 18S rRNA PCR primer pairs
confirmed identities (99 percent or greater, with expect value 2e-97) with Fusarium species (such as 5 JM-2013 isolate
CJBB24-27 and Fusarium solani ) 18S rRNAs (top bold). Bottom panel depicts alignment of sequence of PCR product (blue
letters designated as query) with 18S rRNA sequence of Fusarium solani strain bxq615 rRNA sequence (black letters
designated as subject).
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