Spectrophotometric Studies of Some 2,4-Dinitrophenylhydrazones

Jones, Louis A.; Holmes, J. C.; Seligman, Robert B.

doi:10.1021/ac60110a013

Cited by 180 publications

(78 citation statements)

References 17 publications

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“…Gerbils were injected (i.p.) with either saline or PBN (dissolved in saline) for 14 days prior to sacrifice. Animals were sacrificed by decapitation.…”

Section: Methodsmentioning

confidence: 99%

“…The resulting precipitate was dissolved in 6 M guanidine hydrochloride, and the difference spectrum of the sample treated with DNPH in HCI was determined versus the sample treated with HCl alone. Results are expressed as nmoles of DNPH incorporated per milligram of protein calculated from an extinction coefficient of 21.0 mM-1 cm-l for aliphatic hydrazones (14). GS activity was determined by the method of Rowe et al (15) as modified by Miller et al (16).…”

Section: Methodsmentioning

confidence: 99%

“…In contrast to aged gerbils, PBN treatment of young adult gerbils had no significant effect on brain oxidized protein content or GS activity. Male gerbils, young adults (3 months of age) and retired breeders (15-18 months of age), were treated with PBN for 14 days with twice daily dosages of 32 mg/kg. If PBN administration was ceased after 2 weeks, the significantly decreased level of oxidized protein and increased GS and neutral protease activities in old gerbils changed in a monotonic fashion back to the levels observed in aged gerbils prior to PBN administration.…”

mentioning

confidence: 99%

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Reversal of age-related increase in brain protein oxidation, decrease in enzyme activity, and loss in temporal and spatial memory by chronic administration of the spin-trapping compound N-tert-butyl-alpha-phenylnitrone.

Carney

Starke–Reed

Oliver

et al. 1991

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

657

275

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Oxygen free radicals and oxidative events have been implicated as playing a role in bringing about the changes in cellular function that occur during aging. Brain readily undergoes oxidative damage, so it is important to determine if aging-induced changes in brain may be associated with oxidative events. Previously we demonstrated that brain damage caused by an ischemia/reperfusion insult involved oxidative events. In addition, pretreatment with the spintrapping compound N-tert-butyl-a-phenylnitrone (PBN) diminished the increase in oxidized protein and the loss of glutamine synthetase (GS) activity that accompanied ischemia/reperfusion injury in brain. We report here that aged gerbils had a significantly higher level of oxidized protein as assessed by carbonyl residues and decreased GS and neutral protease activities as compared to young adult gerbils. We also found that chronic treatment with the spin-trapping compound PBN caused a decrease in the level of oxidized protein and an increase in both GS and neutral protease activity in aged Mongolian gerbil brain. In contrast to aged gerbils, PBN treatment of young adult gerbils had no significant effect on brain oxidized protein content or GS activity. Male gerbils, young adults (3 months of age) and retired breeders (15-18 months of age), were treated with PBN for 14 days with twice daily dosages of 32 mg/kg. If PBN administration was ceased after 2 weeks, the significantly decreased level of oxidized protein and increased GS and neutral protease activities in old gerbils changed in a monotonic fashion back to the levels observed in aged gerbils prior to PBN administration. We also report that old gerbils make more errors than young animals and that older gerbils treated with PBN made fewer errors in a radial arm maze test for temporal and spatial memory than the untreated aged controls. These data can be interpreted to indicate that oxidation of cellular proteins may be a critical determinant of brain function. Moreover, it also implies that there is an age-related increase in vulnerability of tissue to oxidation that can be modified by free radical trapping compounds.

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“…Gerbils were injected (i.p.) with either saline or PBN (dissolved in saline) for 14 days prior to sacrifice. Animals were sacrificed by decapitation.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Reversal of age-related increase in brain protein oxidation, decrease in enzyme activity, and loss in temporal and spatial memory by chronic administration of the spin-trapping compound N-tert-butyl-alpha-phenylnitrone.

Carney

Starke–Reed

Oliver

et al. 1991

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

657

275

View full text Add to dashboard Cite

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“…The protein sample was then dissolved with 2 ml of 8 M guanidine hydrochloride, 13 mM EDTA, and 133 mM Tris solution (pH 7.4), and the UV absorbance was measured at 365 nm. The results were expressed as moles of DNPH incorporated per protein (mol/mol) based on an average absorptivity of 21.0 mM Ϫ 1 cm Ϫ 1 (10).…”

Section: Protein Carbonylmentioning

confidence: 99%

Protein-bound 4-hydroxy-2-hexenal as a marker of oxidized n-3 polyunsaturated fatty acids

Yamada

Funada

Shibata

et al. 2004

Journal of Lipid Research

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In the present study, to investigate the contribution of n-3 PUFAs in the oxidative modification of protein in vivo, we characterize the covalent binding of 4-hydroxy-2-hexenal (HHE), a potent cytotoxic aldehyde originating from the peroxidation of n-3 PUFAs, to protein and describe the production of this aldehyde in oxidatively modified LDL and in human atherosclerotic lesions. Upon incubation with BSA, HHE was rapidly incorporated into the protein and generated the protein-linked carbonyl derivative, a potential marker of oxidatively modified proteins under oxidative stress. To detect the protein-bound HHE in vivo, we raised monoclonal antibody HHE53 (MAb HHE53) directed to the HHE-modified protein and identified the Michael addition-type HHE-histidine adduct as the major epitope. This antibody reacted with copper-oxidized LDL, suggesting that HHE was produced during the oxidative modification of LDL. In addition, we demonstrated that the materials immunoreactive to MAb HHE53 indeed constituted the atherosclerotic lesions, in which intense positivity was associated primarily with macrophage-derived foam cells. The results of this study suggest that the reaction between oxidized n-3 PUFAs and protein might represent a process common to the formation of degenerative proteins during aging and its related diseases. -Yamada, S., T. Funada, N. Shibata, M. Kobayashi, Y. Kawai, E. Tatsuda, A. Furuhata, and K. Uchida. Protein-bound 4-hydroxy-2-hexenal as a marker of oxidized n-3 polyunsaturated fatty acids. J. Lipid Res. 2004. 45: 626-634.

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“…The resulting precipitate was dissolved in 6 M guanidine hydrochloride and the difference spectrum of the sample treated with DNPH in HCl was determined versus the sample treated with HCl alone. Results are expressed as nmol of DNPH incorporated per mg of protein calculated from an absorbtivity of 21.0 mM-1 cm-1 for aliphatic hydrazones (24). GS activity was determined by the method of Rowe et al (25) as modified by Miller et al (26).…”

mentioning

confidence: 99%

Oxidative damage to brain proteins, loss of glutamine synthetase activity, and production of free radicals during ischemia/reperfusion-induced injury to gerbil brain.

Oliver

Starke–Reed

Stadtman

et al. 1990

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

675

268

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Free radical-mediated oxidative damage has been implicated in tissue injury resulting from ischemia/ reperfusion events. Global cortical ischemia/reperfusion injury to Mongolian gerbil brains was produced by transient occlusion of both common carotid arteries. Protein oxidation, as measured by protein carbonyl content, increased significantly during the reperfusion phase that followed 10 min of ischemia. The activity of glutamine synthetase, an enzyme known to be inactivated by metal-catalyzed oxidation reactions, decreased to 65% of control levels after 2 hr of reperfusion that followed 10 min of ischemia. We also report that the free radical spin trap N-tert-butyl-a-phenylnitrone [300 mg/kg Oxidative inactivation renders GS and other enzymes highly susceptible to proteolysis by proteases such as trypsin, Streptococcus griseus protease (10), and subtilisin (11) barbital-anesthetized animals, a loop of dental floss was placed around each common carotid artery and the free ends were passed through a double-lumen catheter. The catheter was fixed in the dorsum of the neck and glued in place with cyanoacrylate adhesive. Ischemia was induced by closing the carotid artery against the septum of the double-lumen catheter using the loop of dental floss. Release of tension and removal of the dental floss allowed reperfusion to occur. Ischemia/reperfusion was induced as described (3). Reperfusion was allowed for specific times before decapitation.

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Spectrophotometric Studies of Some 2,4-Dinitrophenylhydrazones

Cited by 180 publications

References 17 publications

Reversal of age-related increase in brain protein oxidation, decrease in enzyme activity, and loss in temporal and spatial memory by chronic administration of the spin-trapping compound N-tert-butyl-alpha-phenylnitrone.

Reversal of age-related increase in brain protein oxidation, decrease in enzyme activity, and loss in temporal and spatial memory by chronic administration of the spin-trapping compound N-tert-butyl-alpha-phenylnitrone.

Protein-bound 4-hydroxy-2-hexenal as a marker of oxidized n-3 polyunsaturated fatty acids

Oxidative damage to brain proteins, loss of glutamine synthetase activity, and production of free radicals during ischemia/reperfusion-induced injury to gerbil brain.

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