K. Mertens, D. A. Lowes, N. R. Webster, J. Talib, L. Hall, M. J

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Low zinc and selenium concentrations in sepsis are associated with oxidative damage and inflammation K. Mertens, D.A. Lowes, N.R. Webster, J. Talib, L. Hall, M.J. Davies, J.H. Beattie, H.F. Galley British Journal of Anaesthesia Volume 114, Issue 6, Pages 990-999 (June 2015) DOI: 10.1093/bja/aev073 Copyright © 2015 The Author(s) Terms and Conditions

Fig 1 (a) Zinc content of lysates of endothelial cells cultured in normal medium alone, or with N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) or zinc pyrithione (ZP), or cultured in medium containing Chelex-treated fetal bovine serum (FBS) with 1 or 20 µM zinc for 24 h. *Significantly different from cells in normal medium and #significantly higher than cells cultured with 1 μM zinc (P<0.05, Wilcoxon–Mann–Whitney test). Box-and-whisker-plots show median, and interquartile and full range (n=6). (b) Relative mRNA expression of ZIP2 zinc importer protein in endothelial cells cultured in medium containing Chelex-treated FBS with zinc added at a range of concentrations for 4 days. Individual data points are shown (n=3). (c) Glutathione peroxidase 3 (GPx3) activity in lysates of endothelial cells exposed to lipopolysaccharide (LPS) and peptidoglycan (PepG) in normal medium (control) or a range of selenium concentrations for 24 h. P-value is Kruskal–Wallis. *Significantly higher than control (P<0.05, Wilcoxon–Mann–Whitney test). Results are expressed as the percentage expression of LPS/PepG-treated control cells in normal medium. Box-and-whisker plots show median, and interquartile and full range (n=6). (d) Interleukin-6 (IL-6) concentrations in culture supernatants from endothelial cells exposed for 24 h to LPS/PepG in normal medium (control) or LPS/PepG with zinc pyrithione (ZP) or TPEN pre-treatment. Results are expressed as the percentage expression of LPS/PepG-treated control cells in normal medium. P-value is Kruskal–Wallis. *Significantly higher than control (P<0.05, Wilcoxon–Mann–Whitney test). Box-and-whisker plots show median, and interquartile and full range (n=6). (e) Interleukin-6 concentrations in culture supernatants from endothelial cells exposed to LPS/PepG in normal medium (control) or LPS/PepG and a range of zinc or selenium concentrations for 24 h. Results are expressed as the percentage of LPS/PepG-treated control cells in normal medium. Box-and-whisker plots show median, interquartile and full range (n=6). British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions

Fig 2 Mitochondrial membrane potential (MMP; a) and mitochondrial metabolic activity (b) in endothelial cells exposed to LPS/PepG in normal medium (control) or LPS/PepG and a range of zinc or selenium concentrations for 24 h. Results are expressed as the percentage of LPS/PepG-treated control cells in normal medium. P-values shown are from Kruskal–Wallis test. *Significantly different from control (P<0.05, Wilcoxon–Mann–Whitney test). Box-and-whisker plots show median, interquartile and full range (n=6). British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions

Fig 3 Adenosine triphosphate (ATP) production in endothelial cells exposed to LPS/PepG in normal medium (control) or LPS/PepG and a range of zinc concentrations for 24 h. Results are expressed as the percentage of LPS/PepG-treated control cells in normal medium. P-values shown are from Kruskal–Wallis test. *Significantly different from control (P<0.05, Wilcoxon–Mann–Whitney test). Box-and-whisker plots show median, interquartile and full range (n=6). British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions

Fig 4 Plasma zinc (a) and selenium concentrations (b) and GPx3 activity (c) in critically ill patients without infection (control, n=18) and patients with sepsis (n=20). Box-and-whisker plots show median, interquartile and full range. P-value shown is Wilcoxon–Mann–Whitney test. British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions

Fig 5 Plasma IL-6 (a), interleukin-8 (IL-8; b), and pentraxin 3 (PTX3; c) C-reactive protein (CRP; d), and soluble urokinase-type plasminogen activator receptor (suPAR) concentrations (e) in critically ill patients without infection (control, n=18) and patients with sepsis (n=20). Box-and-whisker plots show median, interquartile and full range (+=outliers). P-value is Wilcoxon–Mann–Whitney test. British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions

Fig 6 Plasma lipid hydroperoxide (LPO; a), myeloperoxidase (MPO; b), thiocyanate (c), protein thiols (d), plasma protein methionine (e), and plasma protein methionine sulphoxide concentrations (f) in critically ill patients without infection (control, n=18) and patients with sepsis (n=20). Box-and-whisker plots show median, interquartile and full range. P-values shown are Wilcoxon–Mann–Whitney test. British Journal of Anaesthesia 2015 114, 990-999DOI: (10.1093/bja/aev073) Copyright © 2015 The Author(s) Terms and Conditions