The Arochlor 1254-induced rat liver S9 was a kind gift from Dr. Costas Ionnides of the University of Surrey U. The MLA assay protocols were obtained from the Genetic Toxicology
Department of GlaxoSmithKline Company (Ware U.
MIT treatment of SH-SY5Y cells as shown in figure 2. Kratom Growing Kit mSE (figure 2. MIT-like compound (based on the analysis described in section 2.
were counted and 2 x 104 cells were transferred onto microscope slides followed by centrifugation (cytospin at 450 rpm for 5 minute). Y in phosphate buffer) for 5 seconds. The excess stain was then drained onto absorbent paper and the slides were transferred into basic solution dye (methylene blue in phosphate buffer) for another 5 seconds. Finally the slides were rinsed briefly in the buffered water (pH 7. The slides were mounted with DPX and microscopic examination was then carried out similarly as described for WrightGiemsa staining procedure.
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- Cell viability was assessed using Trypan blue exclusion
- SH-SY5Y cells which are known to have wild-type p53 have constitutive expression of p53 in the control and lower doses groups
- In this study therefore an attempt was made to characterise the MSE and MIT toxicity by looking at cell cycle distribution
- Cytotoxicity was apparently unaffected by ketoconazole
- As apoptosis could follow various pathways and often vary in different cells (Esposti and McLennan 1998 Hetts 1998) this prompted us to further investigate if other pathways could contribute
- N-acetyl-L-cycteine affords protection against lead-induced cytotoxicity and oxidative stress in human liver carcinoma (HepG2) cells
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Cytometry 13: 795-808. Determining cell stages by flow cytometry. Current Protocols in Cell Biology.
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Dose selection for the Viability and Mutant Frequency (MF) plating were chosen based on the RSG calculation as described in section 3. Treatment groups Conc. C MSE Treatment with S9 (3 hr) 25 20 15 10 5 DMBA Neg. B MSE Treatment without S9 (24 hr) Neg. C 40 30 20 10 5 MMS Cell conc. X 105 8. Relative suspension growth (RSG) 91.
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