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In preliminary studies, diurnal variations in Eh GSH and Eh CySH were too small to explain our results and have peaks that are separated by 6-7 h, suggesting that the differences that we observed were not the result of these variations alone.
[5] Nonstandard abbreviations: AF, atria] fibrillation; CRP, C-reactive protein; DROMs, derivatives of reactive oxidative metabolites; Eh GSH, oxidized to reduced glutathione; [E.sub.h] CySH, oxidized to reduced cysteine; hsCRP, high-sensitivity CRP; IL, interleukin; TNF[alpha], tumor necrosis factor [alpha].
After centrifugation at 500 x g for 5 min, both the pellets and supernatants were derivatized immediately with mbb (as described above) for CYSH and GSH determination.
A Shimadzu HPLC system equipped with a RF-551 fluorometric detector (Shimadzu Scientific Instruments, Inc., Columbia, MD) and a C18 reverse-phase column (Luna, 5 [micro]m; Phenomenex, Torrance, CA) was used for the analysis of CYSH and GSH.
The corresponding changes in intracellular CYSH and GSH in AM and LNC in relation to the saline control are shown in Figure 2.
Figure 3 shows the effects of DEP exposure on the capacity of AM to generate CYSH and GSH.
The ex vivo incubation with cystine increased CYSH production 2-fold, suggesting that these cells have the capacity to take up and convert cystine to CYSH.
Figure 5 shows that DEP exhibited relatively strong affinities toward cystine, moderate absorptive capacity toward CYSH and GSH, but no effect on GSSG.
The results of our study are consistent with the concept that CYSH and GSH may play a dual role in DEP-induced pulmonary immune/inflammatory responses.
To provide more insight into DEP effects on cellular regulation of thiols, AM and lymphocytes from saline and DEP-exposed animals (72 hr after exposure) were isolated and tested for their production of CYSH and GSH (Figures 3 and 4).
These cells exhibited a moderate capacity in the uptake and conversion of cystine to CYSH. Studies by Ishii et al.