S.M. Eggan et al.

SupplementaryMaterials

Human Subjects

For each subject, an independent committee of experienced research clinicians made consensus DSM-IV (American Psychiatric Association, 1994)diagnoses based on information obtained from clinical records and a structured interview conducted with a surviving relative (Glantz and Lewis, 1997). In the schizophrenia group,seven subjects (four in cohort 1 and three in cohort 2) met criteria for schizoaffective disorder, five subjects (two in the cohort 1 and three in cohort 2) died by suicide, 12 subjects (seven in cohort 1 and five in cohort 2) had a history of substance (including alcohol) abuse or dependence, although only six (five in cohort 1 and onein cohort 2) of them met criteria at the time of death. Six subjects with schizophrenia (three in each cohort) had a history of cannabis use; none of these met criteria for abuse or dependence at time of death, but two (450 and 829) had a history of abuse in remission at time of death. In the MDD group, seven subjects died by suicide, seven subjects had a history of substance (including alcohol) abuse or dependence, and five of these met criteria at the time of death. Three MDD subjects had a history of cannabis use; none of these met criteria for abuse or dependence at time of death. Of the normal comparisonsubjects, one subject (987) had a history of post-traumatic stress disorder that was in remission for 39 years at time of death (Tables S1 and S2).

Toxicology revealed positive plasma alcohol levels in three normal comparison subjects (516, 685, and 344; 0.01 – 0.09%) and three MDD subjects (565, 505, and 602; 0.15 – 0.18%). No other substances of abuse, including Δ9-tetrahydrocannabinol or its metabolites, were detected in any of the subjects. Six schizophrenia subjects were off antipsychotic medications at the time of death (537, 9.6 months; 622, 1.2 months, 829, 207, and 450, discontinued for unknown period of time; 234, antipsychotic naïve; Tables S1 and S2).

Photography

Brightfield photomicrographs were obtained with a Zeiss Axiocam camera (Carl Zeiss MicroImaging, Inc, Thornwood, New York), and the brightness and contrast were adjusted in Adobe Photoshop (Adobe Systems Inc, San Jose, California).

Supplementary Figure Legends

Figure S1. Paired analysis of cortical ROD levels of CB1R immunoreactivity in matched pairs of normal comparison subjects and subjects with schizophrenia (closed squares) or schizoaffective disorder (closed diamonds). Markers below the dashed unity line indicate pairs for which the subject with schizophrenia or schizoaffective disorder had lower mean CB1R immunoreactivity than the matched normal comparison subject. Mean values for each subject group are indicated by the star.

Figure S2. (A) Paired analysis of cortical ROD levels of CB1R immunoreactivity in matched pairs of normal comparison subjects and subjects with schizophrenia (closed squares) or schizoaffective disorder (closed diamonds). (B) Paired analysis of cortical ROD levels of CB1R immunoreactivity in matched pairs of MDD subjects and subjects with schizophrenia (squares) or schizoaffective disorder (diamonds). (C) Paired analysis of cortical ROD levels of CB1R immunoreactivity in matched pairs of normal comparison subjects and MDD subjects. In panels A and B, markers below the dashed unity line indicate pairs for which the subject with schizophrenia or schizoaffective disorder had lower mean CB1R immunoreactivity than the matched normal comparison or MDD subject, respectively. In panels B and C, open markers denote MDD with psychotic features. In all panels, mean values for each subject group are indicated by the star.

Figure S3.The effects of psychosis or suicide on CB1R immunoreactivity levels in subjects with MDD. Mean (bar) and individual (open circles) relative optical density values for CB1R immunoreactivity are shown for MDD subjects grouped by the presence of psychotic features (A) or death by suicide (B). Analysis revealed that neither psychotic features nor suicide in MDD subjects affected levels of CB1R immunoreactivity. Numbers in bars indicate the number of subjects with MDD in each category.

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Supplementary Tables

Table S1 Characteristics of Cohort 1

Subject Group1 / Case No. / S/R/A2 / PMI3 / ST4 / Cause of death5 / DSM IV diagnoses
Primary6 Substance7 / Meds
ATOD8 / Cannabis Use9
1 / C
S / 592
533 / M/B/41
M/W/40 / 22.1
29.1 / 120
130 / ASCVD
Accidental asphyxiation / US / N
P / N
N
2 / C
S / 567
537 / F/W/46
F/W/37 / 15.0
14.5 / 124
129 / Mitral valve prolapse
Suicide by hanging / SA / N
N / N
N
3 / C
S / 516
547 / M/B/20
M/B/27 / 14.0
16.5 / 132
128 / Homicide by gun shot
Heat Stroke / SA / N
B; D; O; P / N
N
4 / C
S / 630
566 / M/W/65
M/W/63 / 21.2
18.3 / 114
125 / ASCVD
ASCVD / US / AAR / N
B; D; P / N
N
5 / C
S / 604
581 / M/W/39
M/W/46 / 19.3
28.1 / 118
123 / Hypoplastic coronary artery
Accidental combined drug OD / PS / ADC; OAC / N
B; O; P / N
Y
6 / C
S / 546
587 / F/W/37
F/B/38 / 23.5
17.8 / 128
121 / ASCVD
Myocardial hypertrophy / US / AAR / N
B; P / N
Y
7 / C
S / 551
625 / M/W/61
M/B/49 / 16.4
23.5 / 127
115 / Cardiac tamponade
ASCVD / DS / AAC / N
D; P / N
N
8 / C
S / 685
622 / M/W/56
M/W/58 / 14.5
18.9 / 107
115 / Hypoplastic coronary artery
Right MCA infarction / US / N
N / N
N
9 / C
S / 681
640 / M/W/51
M/W/49 / 11.6
5.2 / 108
113 / Hypertrophic cardiomyopathy
Pulmonary embolism / PS / N
D; P / N
N
10 / C
S / 727
829 / M/B/19
M/W/25 / 7.0
5.0 / 101
82 / Trauma
Suicide by drug OD / SA / ADC; OAR / N
B; O / N
Y
11 / C
S / 575
517 / F/B/55
F/W/48 / 11.3
3.7 / 123
132 / ASCVD
Intracerebral hemorrhage / DS / ADC / N
P / N
N
12 / C10
S / 987
802 / F/W/65
F/W/63 / 21.5
29.0 / 52
87 / ASCVD
Right ventricular dysplasia / SA / ADC; ODR / N
O; P / N
N
Abbreviations by column: 1: C, comparison; S schizophrenia; 2: A, age in years; B, black; F, female; M, male; R, race; S, sex; W, white; 3:PMI, postmortem interval in hours; 4: ST, storage time in months at -80°C; 5: ASCVD, atherosclerotic cardiovascular disease; CO, carbon monoxide; OD, overdose; 6: DS, disorganized schizophrenia; PS, paranoid schizophrenia; PTSD R, post-traumatic stress disorder in remission at time of death; SA, schizoaffective disorder; US, undifferentiated schizophrenia; 7:AAC, alcohol abuse, current at time of death; AAR, alcohol abuse, in remission at time of death; ADC, alcohol dependence, current at time of death; ADR, Alcohol dependence, in remission at time of death; ODC, other substance dependence, current at time of death; ODR, other substance dependence, in remission at time of death; OAR, other substance abuse, in remission at time of death; 8: Meds ATOD; medications at time of death; B, benzodiazepines; D, antidepressants; N, none; O, other medication; P, antipsychotics; 9:Past history of cannabis use; N, none; Y, yes; 10: Post-traumatic stress disorder in remission for 39 years.

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S.M. Eggan et al.

Table S2 Characteristics of Cohort 2

Subject Group1 / Case No. / S/R/A2 / PMI3 / ST4 / Cause of death5 / DSM IV diagnoses
Primary6 Substance7 / Meds
ATOD8 / Cannabis Use9
1 / C
S
M / 250
398
210 / F/W/47
F/W/41
F/W/50 / 5.3
10.3
4.7 / 208
184
214 / ASCVD
Pulmonary embolus
Suicide by drowning / SA
MDD / N
B; O; P
N / N
U
U
2 / C
S
M / 270
131
249 / M/B/62
M/B/62
M/B/57 / 3.3
3.9
4.3 / 205
231
208 / ASCVD
Pneumonia
ASCVD / US
MDDpf / AAR / O
P
O / N
U
N
3 / C
S
M / 304
236
403 / M/W/74
M/W/69
M/W/70 / 17.5
17.0
16.5 / 200
210
183 / Trauma
Thermal burns
ASCVD / RS
MDD / AAR / N
B; O; P
O / N
U
U
4 / C
S
M / 168
207
493 / M/W/77
M/W/71
M/W/73 / 6.2
3.8
8.3 / 220
215
157 / ASCVD
Subdural hematoma
Myocardial infarction / US
MDM / ODC / N
N
D; O / N
U
N
5 / C
S
M / 452
341
564 / F/W/40
F/W/47
F/W/56 / 14.3
14.5
16.6 / 167
196
146 / ASCVD
Suicide by drug OD
Suicide by hanging / US
MDDpf / AAC / N
O; P
B; D; O / N
N
N
6 / C
S
M / 230
153
513 / M/W/33
M/B/24
M/W/24 / 18.2
18.3
13.1 / 211
222
153 / ASCVD
Sucide by drowning
Suicide by hanging / PS
MDDpf / ADR
ADR; ODR
ODC / B; O
O; P
N / N
Y
Y
7 / C
S
M / 420
333
565 / F/B/67
F/W/66
F/W/62 / 19.5
17.9
12.4 / 178
196
146 / Accidental CO intoxication
ASCVD
Suicide by gunshot / US
MDD / AAC; ODR / N
D; O; P
D / N
N
U
8 / C
S
M / 396
408
511 / M/W/41
M/W/46
M/W/43 / 17.5
19.8
17.9 / 184
182
153 / ASCVD
ASCVD
ASCVD / PS
MDD / N
B; D; O; P
O / N
N
N
9 / C
S
M / 344
422
505 / M/W/50
M/W/54
M/W/57 / 6.8
11.0
12.8 / 195
178
155 / ASCVD
ASCVD
Suicide by gunshot / PS
MDD / ADC / N
B; O; P
O / N
Y
Y
10 / C
S
M / 412
466
421 / M/W/42
M/B/48
M/W/44 / 14.2
19.0
16.0 / 181
165
178 / Aortic stenosis
ASCVD
ASCVD / US
MDD / AAR / O
O; P
N / N
U
U
11 / C
S
M / 390
428
228 / F/W/72
F/W/67
F/W/66 / 11.0
9.0
5.4 / 187
176
211 / ASCVD
COPD
Accidental asphyxiation / SA
MDDpf / AAR / O
B; O; P
D; O / N
N
N
12 / C
S
M / 592
450
614 / M/B/41
M/B/48
M/W/39 / 22.1
22.0
19.5 / 141
167
138 / ASCVD
Suicide by trauma
Suicide by CO intoxication / US
MDD / ADR; OAR; ODR / N
N
N / N
Y
Y
13 / C
S
M / 681
234
602 / M/W/51
M/W/51
M/W/56 / 11.6
12.8
11.8 / 129
211
140 / Hypertrophic cardiomyopathy
Cardiomyopathy
Suicide by gunshot / PS
MDD / ADC / N
N
N / N
N
N
14 / C
S
M / 575
597
580 / F/B/55
F/W/46
F/W/58 / 11.3
10.1
11.3 / 144
141
144 / ASCVD
Pneumonia
ASCVD / SA
MDDpf / N
D; O; P
N / N
N
N
Abbreviations by column: 1: C, normal comparison; S, schizophrenia; M, mood disorder; 2: A, age in years; B, black; F, female; M, male; R, race; S, sex; W, white; 3:PMI, postmortem interval in hours; 4: ST, storage time in months at -80°C; 5: ASCVD, atherosclerotic cardiovascular disease; CO, carbon monoxide; COPD, chronic obstructive pulmonary disease; OD, overdose; 6: MDM, mood disorder (depressed) due to a general medical condition; MDD, major depressive disorder; pf, psychotic features; PS, paranoid schizophrenia; SA, schizoaffective disorder; RS, residual schizophrenia; US, undifferentiated schizophrenia; 7:AAC, alcohol abuse, current at time of death; AAR, alcohol abuse, in remission at time of death; ADC, alcohol dependence, current at time of death; ADR, alcohol dependence, in remission at time of death; OAR, other substance abuse, in remission at time of death; ODC, other substance dependence, current at time of death; ODR, other substance dependence, in remission at time of death; 8: Meds ATOD; mediations at time of death; B, benzodiazepines; D, antidepressants; N, none; O, other medication; P, antipsychotics; 9:Past history of cannabis use; N, none; U, unknown; Y, yes.

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References

American Psychiatric Association (1994): DSM-IV. Diagnostic and Statistical Manual of Mental Disorders. Fourth Edition. American Psychiatric Association: Washington, D.C.

Glantz LA and Lewis DA (1997). Reduction of synaptophysin immunoreactivity in the prefrontal cortex of subjects with schizophrenia: Regional and diagnostic specificity. Arch Gen Psychiatry54: 943-952.

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