A and Douglas B. Mitragyna Speciosa – Pc Bali Kratom Capsules (og) some observations on the pharmacology of mitragynine. Apoptosis oncosis and necrosis.
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BMJ 329: 257-258. BMJ 332: 175-176 Weinert T. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae.
Cells treated with both high concentrations of MSE (Fig. A) and cells bali kratom resin pre-treated with NAC appeared similar to Control group. This infers that MSE did not generate ROS which confirmed the earlier finding. With MIT treatment groups (Fig. B) similar findings were clearly seen. NAC appeared no different compared to Control group.
MSE treated SH-SY5Y cells was not established in my preliminary kratom capsules pill experiments further assays were carried out to confirm this finding. The inhibitors used were caspase 3 inhibitor caspase 8 inhibitor caspase 9 inhibitor general caspase inhibitor negative control and doxorubicin as a positive control ( as described in section 5. The positive control doxorubicin confirmed the assay works by showing a highly significant response for apoptosis. Thus this finding supported the notion that there was no involvement of caspase executioner nor caspase initiator activation in cell death induced by high dose MSE. C o N ntr eg ol a (E M tive tO C M SE co H) a C sp. M E C . MS E 5 9 h E 0 G inh .
The cell pellets were then prepared for flow cytometry analysis using PI staining as described in chapter 4 section 4. The cells stained with PI were analysed using BD FacsCalibur flow cytometer. PI was excited at 488 nm and 620 nm emissions. Ten thousand cells were analysed by CellQuest Pro software and the subG1 population representing apoptotic cells were gated manually.
M respectively accompanied the cell death of the cell. However it appears that there was no involvement of the cell cycle protein p53 and the p21 pathway with MSE. This was not the case with MIT. Dose dependant lost of p53 Mitragyna Speciosa – Pc Bali Kratom Capsules (og) and p21 observed at the same concentrations causing cell cycle arrest remains unexplained. The data also suggested that the cell membrane integrity was compromised leading to the loss of cell content possibly through membrane opening or increased membrane permeability.
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Whereas for MIT as shown in previous Mitragyna Speciosa – Pc Bali Kratom Capsules (og) 4 hr incubation time point similar results were observed for both MIT treated groups. These results suggest that caspase 3 and 7 activities were more pronounced in MIT treated cells and are likely not to be involved in the MSE treated cells. SH-SY5Y cells treated with various concentrations of MSE and MIT at A) 4 hr and B) 18 hr incubation time period.
MSE at any time point. This finding supports the previous p53 results. Parallel experiments were carried out to assess the effects of MIT on the expression of p21 protein. In the previous section it was noted that there were no major differences in p53 band intensity over the dose range tested compared to the control group implying that MIT does not induce the loss of protein as seen in the MSE treated cells. As with the p53 effects noted previously MIT had little effect on p21 levels (Fig. P21 levels of MSE treated SH-SY5Y cells at different time points (6 12 24 and 48 hr).