The quantitation of p21 protein is described in section 4. What Type Of Kratom Is Best For Euphoria there was a clear up regulation of p21 protein seen for What Type Of Kratom Is Best For Euphoria the control group at 24 and 48 hours consistent with the upregulation of p53 noted earlier. 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.
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.
This finding is consistent with the result of the previous flow cytometry analysis with PI staining performed in chapter 4 section 4. For MIT treated cells changes of the four populations were not as drastic as MSE treated cells. Q3 and Q4 indicating increased of apoptotic and necrotic cells.
Mitchell et al 1997). In general MSE with or without the presence of metabolic activation (Arochlor 1254 induced rat liver S9) was negative for genotoxic potential. MSE in the presence of S9 turned out to be positive. RTG and also low RSG (24%) prior plating.
The Medical Journal of Australia 166:538-541. CIP1 is induced in p53-mediated G1 arrest and apoptosis. WAF1 a potential of p53 tumor suppression. Cell 75: 817-825. Measuring mitochondrial reactive oxygen species.
The result was generated from a single preliminary experiment. After this preliminary experiment optimisation of the assay was conducted as described in section 5. DCFHDA precipitations seen in the preliminary assay which could interfere with the fluorescence readings.
PNAS 92: 8493-8497. Mechanism of taxol-induced apoptosis in human SKOV3 ovariancarcinoma cells. The cell cycle and programme cell death.
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 euphoric kratom extract treatment with high doses of MSE or MIT. During the ROS study another interesting thai kratom experience observation was made specifically that MSE co-treatment with NAC appeared to protect What Type Of Kratom Is Best For Euphoria the cells from death and that chemicals present in the MSE emphasised this effect. Tsuchiya et al 2002; Thongpradichote et al 1998; Tohda et al 1997). Thongpradichote et al 1998).
CIP1 is induced in p53-mediated G1 arrest and apoptosis. WAF1 a potential of p53 tumor suppression. Cell 75: 817-825. Measuring mitochondrial reactive oxygen species.
In order green indo kratom powder 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 What Type Of Kratom Is Best For Euphoria (Fig. MS table 2.
Studies of initiation and promotion of carcinogenesis by N-nitroso compounds. Apoptosis: the p53 network. Journal of Cell Sciences 116: 4077-4085. DNA doublestrand break repair: from mechanistic understanding to cancer treatment. DNA Repair (Amst.
However one hypothesis that could be proposed is the possibility of the membrane integrity being compromised especially at high dose of treatment or in other words the lost of cell content through membrane opening. In principle in DNA cycle analysis the movement of DNA profiles to the right side of the scale indicates more dye has been taken up. This would be the implication if the pores of the plasma membrane open or if there was a mechanism in which the dyes could diffuse more easily into the cell. Another flow cytometry analysis was carried out in this chapter this time using double staining with Annexin V conjugates-7-AAD to further determine the nature of cell death.
Based on these observations two possibilities are considered: 1) the effect is cell cycle arrest independent of p53 and p21 pathway or 2) the loss of these proteins could be due to the leakage due to the increased membrane permeability or through pore opening. The toxicity What Type Of Kratom Is Best For Euphoria findings noted thus far are consistent with my hypothesis in which the dose is the main factor in determining the level of the cytotoxicity seen. The cytotoxicity events initially seen as cell cycle arrest proceed to cell death with increasing doses of MSE and MIT. My investigations of morphological microscopic examination on three different cell lines showed different modes of cell death.