Supplementary Figure 1: Scatter plot of brain eicosanoid and docosanoid concentrations in adequate and low LA rats treated with i.c.v. ACSF or LPS for two days. There were no significant differences by two-way ANOVA.
Supplementary Table 1: Number of subjects per group for each experiment and documentation of animal or sample loss.
Adequate LA-ACSF / Adequate LA-LPS / Low LA-ACSF / Low LA-LPS14C-AA kineticsa / Starting sample size / 8 / 9 / 7 / 8
Final sample size: / 6 / 5b / 3c / 6
Starting sample size / 8 / 8 / 8 / 8
Eicosanoids / resolving D / Final sample size / 7 or 8d / 8 / 7e / 8
cPLA2 and iPLA2activity / Final sample size / 8 / 8 / 8 / 8
sPLA2activity / Final sample size / 8 / 8 / 7e / 8
COX activity / Final sample size / 6e / 7e / 7e / 6e,f
COX-2 activity / Final sample size / 4f / 4f / 8 / 7f
aSamplesfrom 2 rats per group were not subjected to lipid analysis because they were erroneously injected with opioids by the animal care facility staff.
brats euthanized because they bled at the site of the surgery incision.
cOne low LA-ACSF rat was euthanized during the femoral artery / vein implantation because of a torn blood vessel which led to uncontrollable bleeding. One low LA-LPS rat did not lose consciousness after infusing Nembutal through the vein line following tracer infusion, suggesting that it was detached. This rat was not subjected to high-energy microwave-fixation and was euthanized with Nembutal.
d Data analyzed with or without the outlier rat that may have not been properly microwaved.
eOne rat died after brain surgery due to unknown causes
d Negative enzyme activity value removed.
eSample size below 8 due to negative enzyme activity values that were removed.
f Sample size below 8 because there was no more cytosolic extract left to perform the activity assay. This was the case for total COX and COX-2 in one low LA-LPS subject, 4 adequate LA-ACSF rats for COX-2, and 4 adequate LA-LPS rats for COX-2.
Supplementary Table 2: Log-transformed brain eicosanoid and docosanoid concentrations following 2-day ACSF or LPS infusion to low or adequate LArats
Adequate LA-ACSF / Adequate LA-LPS / Low LA-ACSF / Low LA-LPS / Treatment / Diet / Interactionn=8 / n=8 / n=71 / n=8
Log (PGE2) / -0.05 ± 0.47 / 0.38 ± 0.29* / -0.04 ± 0.20 / 0.08 ± 0.28 / 0.0255 / NS / NS
Log (TXB2) / 0.1 ± 0.1 / 0.1 ± 0.2 / 0.2 ± 0.3 / 0.3 ± 0.3 / NS / NS / NS
Log (8-isoprostane) / 0.05 ± 0.4 / 0.06 ± 0.2 / -0.15 ± 0.3 / -0.03 ± 0.11 / NS / NS / NS
Log (Resolvin D1) / 0.4 ± 0.3 / 0.2 ± 0.2 / 0.3 ± 0.4 / 0.4 ± 0.3 / NS / NS / NS
Data are mean ± SD of n = 7-8 per group. Data were analyzed with two-way ANOVA followed by Bonferroni’s post-hoc test. Asterisk (*) indicates statistical significant compared adequate LA-ACSF group. LA contributed 5.2% and 0.4% of calories in adequate and low LA diets, respectively.
1n=7 because one rat died after brain surgery due to unknown causes.