Involvement of metabolism in cytotoxicity was further assessed by clonogenicity assay using rat liver S9 (induced by Arochlor 1254); toxicity increased 10-fold in both cell lines. Kratom Anxiety best way to make kratom Withdrawal Hurley to determine if cytotoxicity was accompanied by DNA damage the Mouse lymphoma tk gene mutation assay was used. The results were negative for both MSE and MIT.
Finally thanks to Almighty Allah S. T for showering His blessing giving me strength and patience during hard times and for this amazing opportunity in my life. STATEMENT OF ORIGINALITY I certify that this thesis and the research to which it refers are the production of my own work and that any ideas or quotations from the work of other people published or otherwise are fully acknowledged in accordance with the standard referencing practices of the discipline.
Microscopic technique may also be used to study the detailed morphology of cell death (apoptosis) by using electron microscopy (Odaka and Ucker 1996). Other common techniques to identify apoptosis use specific immunochemical labelling and proceed with microscopic examination include TUNEL assay (terminal deoxynucleotidyl transferase dUTP nick end labeling) (Negoescu et al 1998). The trypan blue exclusion assay using trypan blue dye is a reliable inexpensive and common test for viability (Puranam and Boustany 1998; Perry et al 1997). The principle of using this dye is that viable cells will exclude the dye and remain clear or white whereas the non-viable cell will take up the dye and thus stain blue when visualised under microscopic examination. The cells which have lysed plasma membrane such as in late apoptosis
are permeable to dye (Puranam and Boustany 1998). FITC kratom tincture addiction (fluorescein isothiocyanate) or PI (Vermes et al 1995) or 7-AAD (7Amino-actinomycin D) (Schmid et al 1992).
Effect of MSE on cytotoxicity (A) and
proliferation (B) of HepG2 cells after 24 hr of treatment. The enzymatic reaction (LDH activity) was determined by fluorescence with an kratom best brand excitation wavelength of 560 nm and emission wavelength of 590 nm. Values are means of triplicates. Bars are standard error of the mean (SEM). To assess the effect of MSE on cell proliferation and viability the Trypan Blue exclusion assay was performed. This assay could be used with much higher concentrations of MSE and showed dose and time-dependency in cell proliferation and viability.
Recently the potent analgesic effect of plant extract and its dominant alkaloid mitragynine Kratom Anxiety Withdrawal Hurley (MIT) were confirmed in vivo and in vitro. MIT or similar compounds could be promising alternatives for future pain management treatments. However the potential cytotoxicity of this plant is mitragyna speciosa 250mg unknown. Therefore the
cytotoxicity of methanol-chloroform extract (MSE) and MIT on human cell lines (HepG2 HEK 293 MCL-5 cHol and SH-SY5Y cells) has been examined.