Effect of MSE and MIT on p53 protein levels SH-SY5Y a neuroblastoma cell known to have wild type p53 (Moll et al 1995 1996) was examined by immunoblotting as described in section 4. Image J version 1. The effects of MSE on p53 expression levels were assessed. Kratom Powder Consumption the p53 protein level was found to be decreased in a dose-dependant manner especially at lower
best kratom stores concentrations of MSE treatment for 24 hr as shown in fig. Further experiments were carried out to determine the time course of the down regulation or loss of p53 (Fig. MSE and control groups implying that this cell line expresses p53 protein and the lost of p53 protein seen at high doses was due to treatment effects.
These results indicate that MSE is being activated to a metabolic product that is cytotoxic to both cell lines; however the SH-SY5Y cells appear to be most susceptible. Clonogenicity assay of MSE with rat S9 treated A) SH-SY5Y and B)HEK 293 cells for 24 hr with MSE in the presence of Arochlor 1254-induced rat liver s9. ANOVA with Tukey-Kramer post test.
Arochlor 1254 rat liver S9-mix was used as the exogenous metabolising system and was prepared freshly on the day of the assay. The S9-mix was prepared by mixing 1 part of S9 with 9 parts of co-factor (5. M NADP (Na2) and 27.
Antracyclines induce calpaindependanttitin proteolysis and necrosis in cardiomyocytes. Genetic toxicity assessment: Employing the best science Kratom Powder Consumption for human safety evaluation Part IV: A strategy in genotoxicity testing in drug development: Some examples. Toxicological Sciences 98:39-42 Lu W.
Antracyclines induce calpaindependanttitin proteolysis and necrosis in cardiomyocytes. Genetic toxicity buy kratom overnight assessment: Employing the best science for human safety evaluation Part IV: A strategy in genotoxicity testing in drug development: Some examples. Toxicological Sciences 98:39-42 Lu W.
Models of reactive oxygen species in cancer. Drug Discov Today Dis Models 4: 67-73. F Lai C.
MSE and MIT. From these estimates it appears that the SH-SY5Y cells are the most sensitive of those examined
<img mitragyna speciosa use in the northern states of malaysia src=’http://www.herbalaffiliateprogram.com/herbalsmokeshop/images/natural-e2.jpg’ alt=’Kratom Powder Consumption’>
to the cytotoxic and possibly cytostatic effect of MSE. Based upon my estimation of 42% MIT-like compound in MSE extract the SHSY5Y cell IC50 for is kratom legal for minors MSE is equal to 9. M MIT-like compound. This is not dissimilar to the experimentally determined IC50 for pure MIT of 7. To assess the long-term effect of MSE on surviving cells after acute treatment a clonogenicity assay was performed after 24 hr treatment on HEK 293 and SHSY5Y cells.
MS E 5 9 h E 0 G inh . M 1 0 e G n. M SE 0 en nh S 5 .
Investigation of the possible role of metabolic involvement in the toxicity of MSE The effect of possible involvement of metabolism was investigated using post mitochondrial supernatant S9 from rat liver induced by Arochlor 1254 a kind gift from Prof. Costas Ionnides of University of Surrey U. MSE with or without S9 (8.
Anti-oxidant N-acetyl-L-cysteine (NAC) (5mM) was also added to appropriate wells. 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 subdued lighting and NAC was also added to appropriate wells. C (5% CO2) for 30 minutes.