Volume 66, Issue 2, Pages (August 2004)

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Volume 66, Issue 2, Pages 841-848 (August 2004) Intravenous iron administration induces oxidation of serum albumin in hemodialysis patients Makoto Anraku, Kenichiro Kitamura, Ayumi Shinohara, Masataka Adachi, Ayaka Suenaga, Toru Maruyama, Kei Miyanaka, Taku Miyoshi, Naoki Shiraishi, Hiroshi Nonoguchi, Masaki Otagiri, Kimio Tomita Kidney International Volume 66, Issue 2, Pages 841-848 (August 2004) DOI: 10.1111/j.1523-1755.2004.00813.x Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 1 High-performance liquid chromatography (HPLC) profile of in vitro–oxidized serum albumin. HPLC profile of albumin from a normal subject under basal condition (A). Serum was incubated with 200 μmol/L of H2O2 in the presence or absence of 180 μmol/L of saccharated ferric oxide for 1 hour (B and D) or 24 hours (C and E). Five-μL aliquots of each serum were analyzed by HPLC using a Shodex Asahipak ES-502N column. HMA; mercaptalbumin (reduced form), HNA-1; nonmercaptalbumin (disulfide form), HNA-2; nonmercaptalbumin (oxidized form). Kidney International 2004 66, 841-848DOI: (10.1111/j.1523-1755.2004.00813.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 2 High-performance liquid chromatography (HPLC) profile of serum albumin from HD patients with or without intravenous iron administration (IVIR). Five-μL aliquots of serum from HD patients with or without IVIR, and healthy subjects were subjected to HPLC analysis using a Shodex Asahipak ES-502N column. A representative tracing of HPLC profiles of serum albumin from normal subjects (A). A representative tracing of HPLC profiles of serum albumin from HD patients without IVIR (B). A representative tracing of HPLC profile of serum albumin from patients treated with IVIR (C). Kidney International 2004 66, 841-848DOI: (10.1111/j.1523-1755.2004.00813.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 3 Carbonyl content of major plasma proteins from normal subjects and HD patients with or without intravenous iron administration (IVIR). Plasma samples from HD patients with or without IVIR were derivatized with DNP and subjected to duplicate SDS-PAGE gels (A). Following electrotransfer, one blot was stained with Coomassie brilliant blue G for protein (upper blot) and the second blot was stained for DNP using OxiBlot kit reagents (lower blot). Carbonyl formation of major plasma proteins (albumin, transferrin, immunoglobulin, and fibrinogen) were determined as densitometry ratio of DNP area and protein area, and are reported in densitometry units (B). Values are expressed as mean ± SE; N = 11 patients per group. *P < 0.05 as compared with plasma from patients without IVIR. Kidney International 2004 66, 841-848DOI: (10.1111/j.1523-1755.2004.00813.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 4 Relationship between plasma protein carbonyl content and f(HNA-1) (A) or f(HNA-2) (B). Plasma protein carbonyl contents and f(HNA-1) or f(HNA-2) values from normal subjects and HD patients with or without intravenous iron administration (IVIR) are plotted, and the line shows linear regression of plasma protein carbonyl content and f(HNA-1) or f(HNA-2) values (N = 33, R = 0.674 and R = 0.724, respectively, P < 0.01). Kidney International 2004 66, 841-848DOI: (10.1111/j.1523-1755.2004.00813.x) Copyright © 2004 International Society of Nephrology Terms and Conditions