A modification of the procedure of ROS detection in is smoking kratom as effective 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 allowing ROS to be detected. Interesting observations made at the end of 1 hr incubations of the cells informed that the control cells kratom legal uk 2012 for both MSE and MIT treated experiments become rounded and floating implying that the cells are probably dying perhaps due to lack of nutrient.

Effects of higher dose of MSE on the cell cycle distribution of MCL-5 after 48 hr treatment. Bali Kratom High Milnesand mSE on the cell cycle distribution of MCL-5 cells at different time points (4 8 24 48 72 and 96 hr treatment). Human neuroblastoma- SH-SY5Y cells The effects of MSE and MIT on the cell cycle of SH-SY5Y cells were also determined.

CYP 2E1 inducers for example alcohol. If time had permitted

the role of metabolism in activating MSE and MIT would have been an important area to pursue. As part of a toxicological assessment genotoxic potential of a compound is important to Bali Kratom High Milnesand characterise. A genotoxic agent is capable of causing DNA damage and if repair is unsuccessful it can lead to further major problems such as Bali Kratom High Milnesand carcinogenesis.

After 24 hr incubation the medium was aspirated and the cells were washed with PBS. Digital photographs were taken of each well at magnification x400. Two pictures were taken for each well as indicated in the figure 2 above. The medium was replaced and the cells were treated again as before and returned to incubator.

Free Radic Biol. kratom withdrawal depression Adulterants in herbal products can cause poisoning. British Bali Kratom High Milnesand Medical Journal 313: 117. Long-term mutagenicity Bali Kratom High Milnesand studies with chloroform and dimethylnitrosamine in female lacl transgenic B6C3F1 mice. Mutagen 31: 248-256. Apoptosis-inducing factor (AIF): key to the conserved caspase-independent pathways of cell death?. Evaluation of triacetyloleandomycin alpha-naphtoflavone and dietyldithiocarbamate as selective chemical probes for inhibition of euphoria kratom usa human cytochrome P450.

It is proposed that despite taking up the trypan blue dye the cells were still alive but may not be fully functional. It is speculated that one effect of the MSE treatment could be opening of membrane pores to allow the dyes to get in without proceeding to cell death. However at higher dose of MSE dye uptake is more likely to represent cell death. The 1H-NMR analysis of MSE and MIT from two different sources revealed the similarity of most spectral peaks for both samples of MIT except there is an extra minor peak at 7. MIT from Japan. This contamination was not seen in the MIT from Malaysia.

The fluorescence readings were then taken every 10 minutes interval up to 1 hr as described earlier. Trypan blue exclusion and clonogenicity assays were employed in this study. The trypan blue assay employed for this study was performed as described in chapter 2 section 2.

Planta Medica 60: 580581. Mutational specificity of aflatoxin B1. Comparison of in vivo hostmediated assay with in vitro S9 metabolic activation.

The SH-SY5Y cells

were again used in this assay and the caspase inhibitors purchased from Calbiochem included Caspase-3 inhibitor II (Z-DEVD-FMK) Caspase-8 inhibitor II (Z-IETD-FMK) Caspase-9 inhibitor I (Z-LEHD-FMK) Caspase general inhibitor I (Z-VAD-FMK) negative control (Z-FA-FMK) and positive Bali Kratom High Milnesand control doxorubicin HCL. M of each inhibitor 30 minutes prior to adding the MSE. C (5% CO2) for 48 hr time period. After incubation the cells were harvested and trypsinised as described in chapter 2 section 2. The cell pellets were then prepared for flow cytometry analysis using PI staining as described in chapter 4 section 4.