Life Sciences 74: 2143-2155. Thai Reserve Kratom Maynard detection of carcinogens as mutagens: Bacterial tester strains with R factor plasmids. PNAS 72: 979-983.
MSE) fewer cells remained with the majority of them apoptotic with typical chromatin condensation appearance. For the HEK 293 treated cells (Fig. SH-SY5Y cells as discussed previously. SH-SY5Y cells and necrosis in Thai Reserve Kratom Maynard HEK 293 cells. Cytological examination of SH-SY5Y cells after 48 hr treatment with Thai Reserve Kratom Maynard MSE (24 hr treatment and 24 hr doubling time). Each photo is representative of 3 similar experiments with the same treatment concentration stained with WrightGiemsa staining.
These current experiments suggest that cell cycle arrest could be an associated event for the toxicity effects seen. In order to assess these effects more fully the well established Modfit software was employed for more detailed cell cycle analysis. In general the DNA profiles for MSE treated MCL-5 cells (Fig. MSE table 2.
C 40 30 20 10 5 MMS Cell conc. X 105 8. Relative suspension growth (RSG) 91.
Since in my present study the apoptotic-like cell death induced by MSE was suggested to be caspasesindependent an investigation looking at generation of ROS in mediating the apoptotic events was carried out. Unfortunately the results in my study showed that there was no ROS generation upon treatment with high doses of MSE or MIT. During the ROS study another interesting observation was made specifically that MSE co-treatment with NAC appeared to protect the cells from death and that chemicals present in the MSE emphasised this effect.
Based on the current findings observed in the present studies it is concluded that the methanol-chloroform extract (MSE) of the Mitragyna speciosa Korth (Kratom) leaves and its dominant alkaloid mitragynine (MIT) have potential to cause cytotoxicity to mammalian cells at high doses kratom tea maker and is possibly harmful to human users. MIT is proposed to be a major contributor to MSE cytotoxicity. The main target system of MSE and MIT cytotoxicity is the central nervous system as shown by sensitivity of neuroblastoma cell lines (SH-SY5Y) throughout the studies. In general MSE and to a lesser extent MIT were found to exert their dose dependant cytotoxicity effects in all human cell lines Thai Reserve Kratom Maynard examined both in acute Thai Reserve Kratom Maynard treatment and also in the longer term as assessed by the clonogenicity assay. M arrest for HEK 293 cells.
Bars are the mean of three experiments with SEM. P53 levels of MSE treated SH-SY5Y cells at different time points (6 12 24 and 48 hr). P53 levels of MIT treated SH-SY5Y cells after 24 hr treatment.
Arochlor 1254 is known to be a potent inducer of wide range of mixed-function oxidase enzymes (Puga and Wallace 1998; Ryan et al 1977). CYP 2E1 may have a role in activating MSE toxicity. CYP 2E1 is an important xenobiotic metabolising enzymes for human and rodents which is expressed in the liver. CYP 2E1 can metabolise various substrates including paracetamol fluoxetin alcohol caffeine and many others (Tanaka et al 2000). CYP 2E1 inducers for example alcohol.
This probably could be due to other chemicals that present in MSE preventing the activation of caspase enzymes. Cell death of SH-SY5Y cells after MSE and MIT appeared to be predominantly via apoptosis based on its morphological appearance however biochemically the results discussed above fail to support a caspase mediating event. As apoptosis could follow various pathways and often vary
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in different cells (Esposti and McLennan 1998 Hetts 1998) this prompted us to further investigate if other pathways could contribute.
Fas)-mediated apoptosis: live and let die. Mitochondrial membrane permeabilization in cell death. Wildtype p53 is a cell cycle checkpoint determinant following irradiation.
Protein concentrations of the cell lysates The kratom dosage guide erowid Thai Reserve Kratom Maynard bicinchoninic assay (BCA) is quick and works in a similar way
to the Lowry method. Smith et al 1985). It is one of the recommended assays for determining protein content of cell lysates used for gel electrophoresis in immunoblotting. BCA protein assay kit (Fig.