Repletion of S-Nitrosohemoglobin Improves Organ Function and
Physiologic Status in Swine Following Brain Death
Basil M. Yurcisin, Tara E. Davison, Syreena M. Bibbs, Bradley H. Collins, Jonathan S. Stamler,
and James D. Reynolds
Journal of Pharmacology and Experimental Therapeutics
SUPPLEMENTAL DIGITAL CONTENT
Supplemental Results
General Physiologic Status
The swine were well ventilated (arterial oxygen saturation stayed essentially constant at
100%) with no treatment effects identified for arterial or venous blood pH or pCO2 though all
animals were trending towards acidosis when the study was terminated. The time course of the
arterial pCO2 and pH changes are presented in Supplementary Figure 1. The average met-Hb
levels in the control and 20 ppm ENO groups never exceeded 1%.
Serum Chemistries
Values for additional serum electrolytes along with urine volume, enzymes and
metabolic markers, hematologic parameters not presented in the main manuscript are
presented in Supplemental Table 1.
For both groups, calcium levels declined and sodium, potassium, and chloride concentrations
increased, accompanied by a significant increase in urine volume indicative of diabetes
insipidus; the magnitude of these increases were not different between the two cohorts. The one
electrolyte variant was magnesium, which increased in the control group (consistent with the
impairment of kidney function described in the main text) but remained stable in the ENO group.
For the protein metabolic markers, albumin declined in both groups but globulin only decreased
in the ENO group, which resulted in a reduction of the albumin/globulin ratio. The ENO group
also displayed statistically-significant rises triglycerides and declines in cholesterol. Blood urea
nitrogen (BUN) concentrations were increased, but only the control group exhibited a significant
increase in serum creatinine (Table 1, main text); by extension only the control group showed
an increase in the BUN/creatinine ratio. Gamma-glutamyl transpeptidase (GGTP) was only
reduced in the control.
With respect to hematologic value, both groups exhibited an increases in red blood cell counts
reflective of hemo-concentration and increased serum osmolarity despite supportive fluid
therapy. Platelet counts declined slightly in the ENO group; the decrease in platelets in the
control group did not reach statistical significance. The numbers of lymphocytes and eosinophils
were highly variable but no significant differences were identified. Basophils were only detected
in one animal.
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Supplemental Digital Content Table 1 Additional Serum Chemistries
Parameter (units)
Control
20 ppm ENO
before
after
before
after
Electrolytes
Calcium (mg/dl)
Phosphorus (mg/dl)
Sodium (mEq/l)
Potassium (mEq/l)
Chloride (mEq/l)
Magnesium (mg/dl)
Urine Volume (ml/h)
Albumin (g/dl)
Globulin (g/dl)
9.3 ± 0.7
10.0 ± 1.4
140 ± 3
4.9 ± 0.6
104 ± 4
1.7 ± 0.2
115 ± 72
3.5 ± 0.4
2.4 ± 0.3
8.4 ± 0.6*
10.4 ± 1.6
156 ± 8*
7.0 ± 1.6*
127 ± 4.3*
2.2 ± 0.4*
570 ± 378*
2.9 ± 0.5*
2.1 ± 0.4
9.6 ± 0.4
8.6 ± 0.7
139 ± 2
4.4 ± 0.3
104 ± 2
1.7 ± 0.2
59 ± 55
3.3 ± 0.3
2.2 ± 0.2
8.7 ± 0.5*
9.2 ± 1.0
147 ± 3*
5.5 ± 0.7*
117 ± 3*
1.7 ± 0.2
515 ± 112*
2.6 ± 0.2*
2.0 ± 0.3*
Metabolic Panel
Albumin:Globulin Ratio
Cholesterol (mg/dl)
Triglycerides (mg/dl)
Total Bilirubin (mg/dl)
Urea Nitrogen (mg/dl)
BUN:Creatinine Ratio
GGTP (U/l)
1.7 ± 0.6
88 ± 20
38 ± 14
0.3 ± 0.2
10 ± 2
6.0 ± 1.4
35 ±11
2.4 ± 3.0
75 ±22
55 ± 23
0.1 ± 0.0
13 ± 4*
6.1 ± 2.1
25 ± 10*
1.5 ± 0.3
72 ± 14
27 ± 12
0.8 ± 1.7
11 ± 2
5.7 ± 1.3
39 ± 17
1.3 ±0.1*
53 ± 14*
50 ± 21*
0.1 ± 0.1
15 ± 5*
8.1 ± 2.3*
29 ± 21
5.9 ± 0.9
233 ± 126
5991 ± 1533
259 ± 197
0±0
6.2 ± 1.0*
205 ± 91
7034 ± 1922
464 ± 238
0±0
6.1 ± 0.6
228 ± 88
6354 ± 2146
222 ± 177
0±0
6.4 ± 0.7
174 ± 86*
5346 ± 2563
211 ± 192
16
Blood Cell Counts
RBC (per mm3)
Platelet Count (per mm3)
Lymphocytes (cell count)
Eosinophils (cell count)
Basophils (cell count)
Serum electrolytes and urine volumes, enzymes and metabolic markers, and blood cell counts
before and 10 h after determination of brain death for the two experimental groups. Values in
the “after” columns that are followed by an asterisk (*) indicate a significant difference from the
pre brain death values (p<0.05). No mean (±SD) value is provided for the post brain death
basophils count in the ENO group because these cells were only detected in one animal. There
was no incidence where the magnitude of any change was different between the two groups.
Abbreviation, ENO, ethyl nitrite; BUN, blood urea nitrogen; GGTP, gamma glutamyl
transpeptidase; RBC red blood cell.
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Supplemental Figure 1
Supplemental Digital Content Figure 1. Additional blood gas data. Mean (± SD) time courses for arterial
pCO2 and pH after the determination of brain death for the untreated (solid line) and ethyl nitrite (ENO)
exposed (small dashes) cohorts. Within each graph, baseline mean (± SD) values are depicted by the
bar and arrows. Ventilation was actively managed to maintain end-tidal CO2 < 35 mm Hg. No treatment
effect differences were identified between the two groups for either parameter.
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