Time course studies using the hypoxia marker pimonidazole showed no staining for pimonidazole at JQ-EZ-05 inhibitor 1 or 2 h in B6C3F1 mice treated with APAP. Staining for pimonidazole was present in the midzonal to periportal regions at 4, 8, 24 and 48 h and no staining was observed in centrilobular hepatocytes, the sites of the toxicity. Subsequent studies with the MPT inhibitor cyclosporine A showed that cyclosporine A (CYC; 10 mg/kg) reduced HIF-1 alpha induction in APAP treated mice at 1 and 4 h and did not inhibit the metabolism
of APAP (depletion of hepatic non-protein sulfhydryls and hepatic protein adduct levels). The data suggest that HIF-1 alpha induction in the early stages of APAP toxicity is secondary to oxidative stress via a mechanism involving MPT. In addition, APAP toxicity is not mediated by a hypoxia mechanism. (C) 2011 Elsevier Inc. All rights reserved.”
“Zoysia tenuifolia Willd. ex Trin. is one of the ASP2215 cell line most popularly cultivated
turfgrass. This is the first report of successful plant regeneration and genetic transformation protocols for Z. tenuifolia using Agrobacterium tumefaciens. Initial calli was induced from stem nodes incubated on a Murashige and Skoog (1962) (MS) medium supplemented with 2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg l(-1) 6-benzyladenine (BA), with a frequency of 53%. Compact calli were selected and subcultured monthly on the fresh medium. Sixty-nine percent of the calli could be induced to regenerate plantlets when the calli incubated on a MS medium supplemented with 0.2 mg l(-1) BA under darkness. For genetic transformation, calli were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301 which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing BMS-754807 ic50 beta-glucuronidase gene (gus-int) as a reporter gene. Following co-cultivation, about 12% of the callus explants produced hygromycin resistant calli
on MS medium supplemented with 2 mg l(-1) 2,4-D, 1 mg l(-1) BA, 50 mg l(-1) hygromycin, 500 mg l(-1) cefotaxime after 8 weeks. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing 0.2 mg l(-1) BA, 50 mg l(-1) hygromycin, and 250 mg l(-1) cefotaxime, and about 46% of the resistant calli differentiated into shoots. Finally, all the resistant shoots were rooted on 1/2 MS media supplemented with 50 mg l(-1) hygromycin, 250 mg l(-1) cefotaxime. The transgenic nature of the transformants was demonstrated by the detection of beta-glucuronidase activity in the primary transformants and by PCR and Southern hybridization analysis. About 5% of the total inoculated callus explants produced transgenic plants after approximately 5 months. The procedure described will be useful for both, the introduction of desired genes into Z. tenuifolia and the molecular analysis of gene function.