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Metabolites were identified by co-elution with standards, and quantification was obtained by integration relative to the internal standard (22). Total Cys and Total GSH were calculated using the equations: [Cys]?+?2[CySS] and [GSH]?+?2[GSSG], respectively. The redox states (Eh) of the thiol/disulfide pools were calculated with the Nernst equation using standard [http://www.selleckchem.com/products/dabrafenib-gsk2118436.html Dabrafenib clinical trial] potentials for the GSH/GSSG (?264?mV) and Cys/CySS (?250?mV) pairs at pH 7.4, as previously described (16, 24). Statistical methods All statistical analyses were performed using SAS 9.3 (Cary, NC, USA) and statistical significance was assessed at the 0.05 significance level unless otherwise noted. Since specific differences in metabolite and redox values are difficult to predict, statistical power was evaluated by utilization of an effect size. Samples of 61 cases and 16 controls were obtained giving 80% power to detect an effect size of 0.8 using a two-sample Wilcoxon-test. Prior to the analysis, the assumption of normality for the metabolite and redox values was assessed using the Shapiro-Wilk test for normality and by visual inspection of the histograms. As has been previously reported (16), evaluation of metabolite data demonstrated that none of the metabolites, with the exception of the redox potentials (Eh), were normally distributed, and thus, values were log transformed and adjusted for differences in age, gender, and race using generalized linear models. Back-transformed [https://en.wikipedia.org/wiki/Binedaline binedaline] least-squares means and 95% confidence intervals are reported. Discrete variables are shown by frequency counts and percentages. Results Subjects and [http://www.selleckchem.com/products/bmn-673.html http://www.selleckchem.com/products/bmn-673.html] clinical characteristics Demographic data are listed in Table ?Table1.1. Primary diagnoses in the category labeled ��Other�� included: severe head trauma, post-operative from spinal fusion surgery, cerebral hypertension, tuberculosis, status epilepticus, brain abscess, post-bronchial foreign body, pertussis, traumatic brain injury, acute chest syndrome, parapneumonic effusion, neutropenic fever, and demyelinating central nervous system disease. Table 1 Demographics, primary diagnosis, and severity of illness measures. Redox status of PICU subjects compared with healthy children Redox data from the PICU subjects and healthy control children are shown in Figure ?Figure1.1. After adjusting for age, gender, and race, children admitted to the PICU had 2.5 times less Cys (2.29 vs. 0.91?��M; p?=?0.009), 2.8 times less CySS (2.98 vs. 1.05?��M; p?=?0.011), and 2.5 times less Total Cys (8.52 vs. 3.41?��M; p?=?0.01) (Figure ?(Figure1A).1A). Children admitted to the PICU had no difference in GSH concentrations (0.29 vs. 0.22?��M; p?=?0.573) or Total GSH (0.37 vs. 0.69?��M; p?=?0.106), but PICU patients had 13 times more GSSG (0.01 vs. 0.13?��M; p?