Mass spectrometric quantification of F2-isoprostanes in biological fluids and tissues as measure of oxidant stress

Jd, Morrow; Lj, Roberts

doi:10.1016/s0076-6879(99)00106-8

Cited by 373 publications

(273 citation statements)

References 7 publications

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“…3B, "pre") was 69 ± 38 pg/mL (range, 30-182 pg/mL). In previous studies using the same GC-MS methodology, a level of 35 ± 6 pg/mL was found for normal human plasma (34), and a value >50 pg/mL was considered elevated in a study involving 396 nonsmoking healthy adults (35). Of the gout patients in the present study, 17 (65%) exceeded the normal mean by 2 SD, and 15 (58%) exceeded the normal mean by 3 SD; the baseline plasma F2-IsoP concentration in these 15 patients ranged from 53-182 pg/mL.…”

Section: )mentioning

confidence: 80%

“…An aliquot of the plasma was acidified for measuring PUA by HPLC (20,31). Additional aliquots of plasma were frozen at −80°C until they were sent on dry ice to Vanderbilt University for measurement of F2-IsoPs by GC negative ion chemical ionization mass spectrometry (34). PCs in plasma were determined according to the manufacturer's instructions using an enzyme immunoassay kit (Biocell PC Test; Biocell Corp, Ltd.).…”

Section: Methodsmentioning

confidence: 99%

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Treating gout with pegloticase, a PEGylated urate oxidase, provides insight into the importance of uric acid as an antioxidant in vivo

Hershfield

Roberts²,

Ganson³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

124

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A high plasma urate concentration (PUA), related to loss of urate oxidase in evolution, is postulated to protect humans from oxidative injury. This hypothesis has broad clinical relevance, but support rests largely on in vitro data and epidemiologic associations. Pegloticase therapy generates H 2 O 2 while depleting urate, offering an in vivo test of the antioxidant hypothesis. We show that erythrocytes can efficiently eliminate H 2 O 2 derived from urate oxidation to prevent cell injury in vitro; during therapy, disulfide-linked peroxiredoxin 2 dimer did not accumulate in red blood cells, indicating that their peroxidase capacity was not exceeded. To assess oxidative stress, we monitored F2-Isoprostanes (F2-IsoPs) and protein carbonyls (PC), products of arachidonic acid and protein oxidation, in plasma of 26 refractory gout patients receiving up to five infusions of pegloticase at 3-wk intervals. At baseline, PUA was markedly elevated in all patients, and plasma F2-IsoP concentration was elevated in most. Pegloticase infusion rapidly lowered mean PUA to ≤1 mg/dL in all patients, and PUA remained low in 16 of 21 patients who completed treatment. F2-IsoP levels did not correlate with PUA and did not increase during 15 wk of sustained urate depletion. There also was no significant change in the levels of plasma PC. Because refractory gout is associated with high oxidative stress in spite of high PUA, and profoundly depleting uric acid did not increase lipid or protein oxidation, we conclude that urate is not a major factor controlling oxidative stress in vivo.

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Section: )mentioning

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Treating gout with pegloticase, a PEGylated urate oxidase, provides insight into the importance of uric acid as an antioxidant in vivo

Hershfield

Roberts²,

Ganson³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

124

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“…Paraformaldehyde-fixed frozen sections were hybridized with 35 S-labeled cRNA probes as described (13).…”

Section: Methodsmentioning

confidence: 99%

Uterine FK506-binding protein 52 (FKBP52)–peroxiredoxin-6 (PRDX6) signaling protects pregnancy from overt oxidative stress

Hirota

Acar

Tranguch

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Immunophilin FK506-binding protein 52 (FKBP52) is a cochaperone that binds to the progesterone receptor (PR) to optimize progesterone (P 4 )-PR signaling. We recently showed that Fkbp52-deficient (Fkbp52 −/− ) mice have reduced uterine PR responsiveness and implantation failure which is rescued by excess P 4 supplementation in a genetic background-dependent manner. This finding led us to hypothesize that FKBP52 has functions in addition to optimizing PR activity. Using proteomics analysis, we found that uterine levels of peroxiredoxin-6 (PRDX6), a unique antioxidant, are significantly lower in Fkbp52 −/− mice than in WT and PR-null (Pgr −/− ) mice. We also found that Fkbp52 −/− mice with reduced uterine PRDX6 levels are susceptible to paraquat-induced oxidative stress (OS), leading to implantation failure even with P 4 supplementation. The same dose of paraquat did not interfere with implantation in WT mice. Moreover, treatment with antioxidants α-tocopherol and N-acetylcysteine (NAC) attenuated paraquat-induced implantation failure in P 4 -treated Fkbp52 −/− mice. Functional analyses using mouse embryonic fibroblasts show that Fkbp52 deficiency associated with reduced PRDX6 levels promotes H 2 O 2 -induced cell death, which is reversed by the addition of NAC or by forced expression of PRDX6, suggesting that Fkbp52 deficiency diminishes the threshold against OS by reducing PRDX6 levels. These findings provide evidence that heightened uterine OS in Fkbp52 −/− females with reduced PRDX6 levels induces implantation failure even in the presence of excess P 4 . This study shows that FKBP52-PRDX6 signaling protects pregnancy from overt OS.embryo implantation | mouse | uterus

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“…Selenium was measured in serum by graphite furnace atomic absorption spectrometry at 196.0 nm in a procedure based on the methods described by Paschal and Kimberly (1986). Measurement of 15-F 2t -IsoP isoprostane, was by a negative chemical ionization GC/MS method modified from that of Morrow and Roberts (1999;Parker et al, 2001). Samples for individual subjects were grouped in random order for analysis and were analyzed within the same batch.…”

Section: Laboratory Methodsmentioning

confidence: 99%

Moderate alcohol consumption and levels of antioxidant vitamins and isoprostanes in postmenopausal women

et al. 2004

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Background: Although alcohol intake has been positively associated with breast cancer risk in epidemiologic studies, the mechanisms mediating this association are speculative. Objective: The Postmenopausal Women's Alcohol Study was designed to explore the effects of moderate alcohol consumption on potential risk factors for breast cancer. In the present analysis, we evaluated the relationship of alcohol consumption with antioxidant nutrients and a biomarker of oxidative stress. Design: Participants (n ¼ 53) consumed a controlled diet plus each of three treatments (15 or 30 g alcohol/day or a no-alcohol placebo beverage), during three 8-week periods in random order. We measured the antioxidants, vitamin E (alpha (a)-and gamma (g)-tocopherols), selenium, and vitamin C in fasting blood samples which were collected at the end of diet periods, treated and frozen for assay at the end of the study. We also measured 15-F 2t -IsoP isoprostane, produced by lipid peroxidation, which serves as an indicator of oxidative stress and may serve as a biomarker for conditions favorable to carcinogenesis. Results: After adjusting for BMI (all models) and total serum cholesterol (tocopherol and isoprostane models) we observed a significant 4.6% decrease (P ¼ 0.02) in a-tocopherol and a marginally significant 4.9% increase (P ¼ 0.07) in isoprostane levels when women consumed 30 g alcohol/day (P ¼ 0.06 and 0.05 for overall effect of alcohol on a-tocopherol and isoprostanes, respectively). The other antioxidants were not significantly modified by the alcohol treatment. Conclusions: These results suggest that moderate alcohol consumption increases some biomarkers of oxidative stress in postmenopausal women. Sponsorship: NCI, NIH and ARS, USDA.

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Mass spectrometric quantification of F2-isoprostanes in biological fluids and tissues as measure of oxidant stress

Cited by 373 publications

References 7 publications

Treating gout with pegloticase, a PEGylated urate oxidase, provides insight into the importance of uric acid as an antioxidant in vivo

Treating gout with pegloticase, a PEGylated urate oxidase, provides insight into the importance of uric acid as an antioxidant in vivo

Uterine FK506-binding protein 52 (FKBP52)–peroxiredoxin-6 (PRDX6) signaling protects pregnancy from overt oxidative stress

Moderate alcohol consumption and levels of antioxidant vitamins and isoprostanes in postmenopausal women

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