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Page 1 of 1 1 (3 articles found)
G Fechner,F G E Perabo,D H Schmidt,L Haase,E Ludwig,H Schueller,J Blatter,S C Mller,P Albers OBJECTIVES: Despite clinical use, the radiosensitizing effect of gemcitabine (2'2'-difluorodeoxycytidine) in human transitional cell carcinoma (TCC) has not been shown to date. We investigated gemcitabine as a radiosensitizer for human TCC cells. METHODS: Monolayer cultures of RT112 (G1, p53 wild t... ( view more )ype), RT4 (G1-G2, p53 wild type), T24 (G3, p53, mutant type), and SUP (G4, p53 mutant type) cells were incubated in medium with gemcitabine. Electron beam radiation was applied alone, simultaneous, or 3, 6, 12, and 24 hours after gemcitabine. Jurkat leukemia cells were used as controls for radiation toxicity. Cell survival was determined 6, 12, 24, 48, and 72 hours after radiation by microculture tetrazolium assay. DNA damage was evaluated by flow cytometric assessment of poly(ADP-ribose) polymerase, and apoptosis was determined by terminal-deoxynucleotidyltransferase-mediated dUTP nick-end labeling and flow cytometric assessment after annexin-V and propidium iodide labeling. RESULTS: In all TCC cell lines, radiation alone caused only little and insignificant growth inhibitory effects at 10 Gy. Gemcitabine alone had a dose-dependent cytotoxic and apoptosis inducing effect on all TCC cell lines independent of p53 status. Assays combining radiation with gemcitabine in different dose and time schedules demonstrated no radiosensitizing effect in TCC cells. CONCLUSIONS: Gemcitabine is effective in TCC cell lines independent of p53 status. A radiosensitizing effect could not be demonstrated. Again, p53 status was not predictive of the radioresponse in the bladder cancer cell lines. Clinical studies with gemcitabine and radiotherapy might nevertheless yield different results but should be performed with utmost caution. ( view less ) A H Buist,M Mller,R I Ghauharali,G J Brakenhoff,J A Squier,C J Bardeen,V V Yakovlev,K R WilsonBy varying the chirp of high-intensity pulses, we can use the chirp-condition-dependent fluorescence yield to distinguish among different molecules or the same molecule in different microenvironments. As an example of the latter we show that SNAFL-2, a well-known pH-sensitive dye, shows large modul... ( view more )ation in fluorescence yield in response to both variation in acidity and variation in chirp condition. Future application of this technique as a novel contrast mechanism within fluorescence microscopy is discussed. ( view less ) G Appanna,B K Tang,R Mller,W Kalow Previous studies have shown that bupranolol, a beta-adrenoceptor blocker, is a substrate of cytochrome P4502D6 (CYP2D6). A sensitive in vitro assay was developed to quantify the formation of hydroxybupranolol using HPLC. A TLC method, using radiolabeled bupranolol, was also developed to test the re... ( view more )producibility of the two methods. Both of them gave virtually identical results; however, the HPLC method was sensitive to 20 pmol and the TLC with radiolabeled substrate to <1 pmol of hydroxybupranolol. The KM value for bupranolol was lower than that reported for any other substrate of CYP2D6. The KM value in microsomes of a typical human liver (L-1) was 0.272 +/- 0.02 (SE) mu M and the Vmax was 360 +/- 10 (SE) pmol/mg/min (0.83 +/- 0.02 pmol/pmol cytochrome P450/min). The KM value for the CYP2D6 expressed in yeast was 0.076 +/- 0.003 (SE) mu M, and the Vmax was 43 +/- 1 (SE) pmol/mg/min (0.64 +/- 0.01 pmol/pmol cytochrome P450/min). Quinidine competitively inhibited the formation of hydroxybupranolol, with Ki values of 5 nM in expressed CYP2D6 and 14.05 nM in human liver (L-1). ( view less )
Page 1 of 1 1 (3 articles found)
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