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Thus it is suggested that apart from MIT there are other chemicals present in the leaves of Mitragyna specioa Korth contributing to the MSE cytotoxicity. A summary of the cytotoxic events leading to MSE or MIT induced SH-SY5Y cell death as discussed above are shown in fig. Mechanisms of MSE and MIT induced mitragyna speciosa cambodia prospect SH-SY5Y cells arrest and cell death.

Nat Rev Cancer. Kratomherbs Lake Mary Fl death receptor: signalling and modulation. Science 281: 1305- 1308. Opioid receptors and legal highs: Salvia divinorum and Kratom. Clinical Toxicology 46: 146-152.

Effects of MSE on the cell cycle distribution of SH-SY5Ycells after 48

hr of treatment. MSE on the cell cycle distribution of SH-SY5Y cells at different time points (4 8 24 48 72 and 96 hr treatment). Indicates only one experimental result. Effect of MIT on cell cycle distribution of SH-SY5Y cells after 24 hr treatment. Histograms and values of the cell cycle phases are kratom how long before withdrawal representative of a single experiment analysed by Modfit software. Protein concentrations of the cell lysates The bicinchoninic assay (BCA) is quick and works in a similar way to the Lowry method. Smith et al 1985).

Measurement of cll-cylce phase-specific cell death using Hoechts 33342 and propidium iodide: Preservation by ethanol fixation. The Journal of Kratomherbs Lake Mary Fl Histochemistry and Cytochemistry 36:1147-1152. Laboratory procedure for assessing specific locus mutations at the TK locus in cultured L5178Y mouse lymphoma cells.

Another important microscopic observation was made after the final redings at the 1 hr time point which kratom capsules speciosa showed that all cells in the Control group appeared rounded and floating in the middle of the well. Similar observations were also noted for H202 MSE and MIT groups. Interestingly the majority of the cells which were treated with NAC prior to treatment with H202 appeared firmly attached to the bottom of the wells and had normal cell appearance. Brownish precipitations were also noted floating in all wells believed to
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be the hydrophobic fluorescent dye DCFH-DA.

Relative suspension growth (RSG) 91. Y Y Y Y Y Y Y Y Y Y Y Conc. Summary table of MLA result for MSE in the i) presence of S9 and ii) in the absence of S9. S9 treatment Treatment groups Negative control MSE 0 0 0 40 30 20 Positive control (MMS) Mean Cotrol MF 75. Negative Negative Negative Negative Negative Negative Positive Conc.

MSE were observed and mechanisms other than direct genotoxicity per se can lead to false

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positive results which are related to cytotoxicity and not genotoxicity such as events associated with apoptosis etc (ICH 1995). Such events are likely to happen once a certain concentration threshold is reached for a toxic compound. For instance in figure 2. A in the previous chapter (section 2. MSE in the presence of S9 reduced the colony formation to less than 10% of the vehicle treated control. A similar outcome was seen using S9 with L5178Y cells in this assay in the preliminary tests for selecting the range of concentrations performed prior to plating assessment.