Table 3.
Standardization of the RBH assay by control proteins and indicative samples, and vs the ntrDNPH assay.
|
Assay | |||
|---|---|---|---|
| Samples | RBH | ||
| nmoles mg−1 protein | |||
| Control proteins | |||
| BSAoxa | 22.9 ± 1.8 | ||
| BSAreda | ~ 0 | ||
| Indicative sample proteinsb: cytoplasmic / histone (ratio) | |||
| Cauliflower B. oleracea (CBo) | |||
| CBountr | 9.3 ± 0.79 / 1.7 ± 0.11 (5.5) | ||
| CBored | ~ 0 | ||
| Lettuce L. sativa (LLs) | |||
| LLsuntr | 24.9 ± 1.9 / 1.0 ± 0.09 (25) | ||
| LLsred | ~ 0 | ||
| Rat Brain Stem (RBS) | |||
| RBSuntr | 0.62 ± 0.05 / 0.60 ± 0.07 (1) | ||
| RBSred | ~ 0 | ||
| Rat Intestine (RI) | |||
| RIuntr | 0.71 ± 0.09 / 0.56 ± 0.06 (1.3) | ||
| RIred | ~ 0 | ||
| Human blood protein carbonyls | |||
| Serum carbonylsc | FTC assayf | ||
| Assays | |||
| Subjectsc | RBH | ntrDNPHc | |
| Males | 1.08 ± 0.07c, d, e | 1.09 ± 0.07c, d, e | ~ 0.43 [17] |
| Females | 1.14 ± 0.09c, d, e | 1.17 ± 0.09c, d, e | |
IMPORTANT NOTE: The present study did not perform the FTC assay because the assumed by the assay stoichiometry 1:1 for the protein carbonyl-FTC hydrazone and the FTC reagent (used to construct the assay's standard curve [17]) is not supported by the referred literature [14].
Control lysozymeox/red, pepsinox/red were also tested and gave analogous results (data not shown).
Values for cytoplasmic and histone protein fractions are shown by the numbers left and right to the separating slash, respectively, while the values in parenthesis represent their cytoplasmic/histone carbonyl content ratio. Histone carbonyls were not detected by the much lower in sensitivity ntrDNPH assay [1]. Values are averages from at least 5 independent measurements (SD, not shown, is < 10% of the average).
Values (in nmoles mg−1) are averages (± SD) from 10 male and 10 middle age female middle age subjects. Values by the ntrDNPH assay are from previous study [1].
Comparison of the new assays for the same and between sex; there is no statistical difference for values statistically significant at p < 0.05.
Comparison between males and females for the three assays; there is a ~ 30% statistical difference for the stdDNPH assay for values statistically significant at p < 0.05.
Carbonyl values determined by the FTC assay (on plasma from unspecified gender) are listed for comparison with the RBH assay.