BACKGROUND: To assess the redox state in hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients, we focused on the formation of glutathionyl hemoglobin (Hb) because the ratio of oxidized glutathione disulfide (GSSG) to reduced glutathione (GSH) is increased in uremia, and GSSG is a source of glutathionyl Hb. METHODS: Glutathionyl Hb levels were measured in 30 HD patients, 10 CAPD patients, and 20 healthy subjects by using liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). RESULTS: Hbbeta showed a peak at 15,868 D in a deconvoluted ESI mass spectrum. Glutathionyl Hbbeta was detected at 16,173 D (15,868 + 305). The peak at 16,173 D was identified as glutathionyl Hbbeta based on the following findings: (1) the peak disappeared by reducing the sample with dithiothreitol, and (2) the peak could be detected at a high level by incubating Hb in vitro with GSH in water at 37 degrees C for seven days. Glutathionyl Hb levels expressed as the peak height ratios of glutathionyl Hbbeta to intact Hbbeta were significantly elevated in HD patients (8.0 +/- 3.6%, mean +/- SD, N = 30, P < 0.0001) and CAPD patients (5.9 +/- 2.7%, N = 10, P < 0.05) as compared with normal subjects (3.0 +/- 1.6%, N = 20). However, there were no significant differences in the glutathionyl Hb levels before (8.7 +/- 3.2%, N = 12) and after HD (8.7 +/- 2.8%, N = 12). CONCLUSION: Glutathionyl Hb levels were increased in HD and CAPD patients, probably because of enhanced oxidative stress. The measurement of glutathionyl Hb may be useful to assess oxidative stress in uremic patients.
BACKGROUND: To assess the redox state in hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients, we focused on the formation of glutathionyl hemoglobin (Hb) because the ratio of oxidized glutathione disulfide (GSSG) to reduced glutathione (GSH) is increased in uremia, and GSSG is a source of glutathionyl Hb. METHODS:Glutathionyl Hb levels were measured in 30 HDpatients, 10 CAPD patients, and 20 healthy subjects by using liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). RESULTS: Hbbeta showed a peak at 15,868 D in a deconvoluted ESI mass spectrum. Glutathionyl Hbbeta was detected at 16,173 D (15,868 + 305). The peak at 16,173 D was identified as glutathionyl Hbbeta based on the following findings: (1) the peak disappeared by reducing the sample with dithiothreitol, and (2) the peak could be detected at a high level by incubating Hb in vitro with GSH in water at 37 degrees C for seven days. Glutathionyl Hb levels expressed as the peak height ratios of glutathionyl Hbbeta to intact Hbbeta were significantly elevated in HDpatients (8.0 +/- 3.6%, mean +/- SD, N = 30, P < 0.0001) and CAPD patients (5.9 +/- 2.7%, N = 10, P < 0.05) as compared with normal subjects (3.0 +/- 1.6%, N = 20). However, there were no significant differences in the glutathionyl Hb levels before (8.7 +/- 3.2%, N = 12) and after HD (8.7 +/- 2.8%, N = 12). CONCLUSION:Glutathionyl Hb levels were increased in HD and CAPD patients, probably because of enhanced oxidative stress. The measurement of glutathionyl Hb may be useful to assess oxidative stress in uremicpatients.
Authors: Victoria M Harper; Joo Yeun Oh; Ryan Stapley; Marisa B Marques; Landon Wilson; Stephen Barnes; Chiao-Wang Sun; Tim Townes; Rakesh P Patel Journal: Antioxid Redox Signal Date: 2014-11-10 Impact factor: 8.401
Authors: David C Ehrmann; Kristie Rose; M Wade Calcutt; Amy B Beller; Salisha Hill; Theresa J Rogers; Steven D Steele; David L Hachey; Judy L Aschner Journal: J Mass Spectrom Date: 2014-02 Impact factor: 1.982
Authors: Dario A Vitturi; Chiao-Wang Sun; Victoria M Harper; Bessy Thrash-Williams; Nadiezhda Cantu-Medellin; Balu K Chacko; Ning Peng; Yanying Dai; J Michael Wyss; Tim Townes; Rakesh P Patel Journal: Free Radic Biol Med Date: 2012-11-16 Impact factor: 7.376
Authors: Khaled Khazim; Daniela Giustarini; Ranieri Rossi; Darlene Verkaik; John E Cornell; Sue E D Cunningham; Maryam Mohammad; Kara Trochta; Carlos Lorenzo; Franco Folli; Shweta Bansal; Paolo Fanti Journal: Transl Res Date: 2013-01-17 Impact factor: 7.012