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Clonogenicity of A) HEK 293 cells and B) SH-SY5Y cells after 24 hr treatment with MSE. MIT treatment of SH-SY5Y cells as shown in figure 2. Pimpernel Bird Placemats mSE (figure 2. MIT-like compound (based on the analysis described in section 2.

The DNA profiles of SH-SY5Y cells were also assessed after exposure to various concentrations of MIT at 24 hr treatment period (Fig. M MIT where cells accumulated at G1 phase and the population shifted to the right side of the scale. This phenomenon implies that the treated cells have taken up more PI dye thus leading to a shift to the right. Due to the amount of MIT compound available repetition of this experiment was not possible.

A great number of studies have demonstrated that central execution of apoptosis by mitochondria can play a critical role in cell death (Esposti and McLennan 1998). The majority of mitochondrial alterations which lead to apoptosis involve an increase of ROS production (Zamzami et al 1995). An example of involvement of ROS production in early stages of apoptosis pathway is provided by ceramide-induced apoptosis (Radin 2001; 2003). A modification of the procedure of ROS detection in live cells adapted from Esposti and McLennan method (1998) was performed; it revealed that both MSE and MIT at high doses did not generate ROS. This result suggests that the mitochondria are still functioning normally or if the MSE and MIT could cause membrane opening or change the membrane permeability the DCFH-DA dye could leak out from cells and thus not buy kratom in ri allowing ROS to be detected. Interesting observations made at the end of 1 hr incubations of the cells informed that the control cells for both MSE and MIT treated experiments become rounded and floating implying that the cells are probably dying perhaps due to lack of nutrient. ultra enhanced indonesian uei kratom la jara Yet co-treatment of cells with NAC prevented this toxicity particularly with MSE.

John Wiley and sons publications. De Vries N. De Flora S. Journal of Cellular Biochemistry supplement 17F: 270-277. Genetic alterations and DNA repair in human carcinogenesis. Safety issues in herbal medicines: implications for the health professions.

Whereas for the longer term effects (clonogenicity assay) fig. M successfully gave protection against MSE borneo red vein crushed kratom toxicity at all dose range however it was not that effective for MIT at high dose. MSE mediates its toxicity via this receptor as shown in acute treatment of MSE (trypan blue exclusion Fig.

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DNA kratom extract or powder doublestrand break repair: from mechanistic understanding to cancer treatment. DNA Repair (Amst. Functions of poly (ADP-ribose) polymerase (PARP) in DNA repair genomic integrity and cell death. Fundamental and Molecular Mechanisms of Mutagenesis 477:97-110. To die or not to die: An overview of apoptosis and its role

in disease.

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

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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 Pimpernel Bird Placemats on p21 levels (Fig. P21 levels of MSE treated SH-SY5Y cells at different time points (6 12 24 and 48 hr). The blots were representatives of duplicate experiments. P21 levels of MIT treated SH-SY5Y cells at different time points (6 12 24 and 48 hr). M for MSE and MIT respectively (Chapter 2).

Fluorescent was measured using a plate reader with 485 nm excitation and 530 nm emission. After 30 minutes cells in each well were treated with H202 MSE and MIT and the fluorescent readings were continually read at 10 min intervals for up to 1 hr period. This preliminary assay was performed to establish the working conditions for the assay. As described earlier the cultured medium was aspirated and fresh PBS (1 ml) was added to each well. M) was then added to the wells under
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subdued lighting and NAC was also added to appropriate wells.

Yano S Horie S. Inhibitory effect of mitragynine an alkaloid with analgesic effect from thai medicinal plant Mitragyna speciosa on electrically stimulated contraction of isolated guinea-pig ileum through the opioid receptor. Life Sciences 60: 933-942. Mitragyna speciosa) a Thai medical plant with special reference to its analgesic activity. In: Tongroach P.

The control and low dose groups however did express p21 protein consistent with the p53 expression. In the parallel experiment with MIT again p21 was expressed in a time-dependant manner that correlated with p53 expression. MIT exerts weaker toxicity effects compared to MSE. Collectively the current findings suggest that MSE induces a cycle arrest that appears to be independent of p53 pathway. In contrast MIT appears to induce cell cycle arrest that is p53 dependant. M respectively accompanied the cell death of the cell.