K) and absorbance was read at 560 nm. Purple Sticky Kratom 15x Extract 3 Grams one set of similar concentrations were also prepared as a negative control (without adding caspase substrate). NA) or caspase -9 (LEHD) substrate were added to the test samples. C prior reading the absorbance at 405 nm using plate reader.
C (5% CO2) for the designated time period. The adherent cells (HEK 293 and SH-SY5Y cells) were harvested trypsinised and centrifuged as per routine procedures described in chapter 2 sections 2. After this incubation the cells were harvested as previously described (section 2. The cell pellets obtained were re-suspended in 1 ml cold PBS or D-PBS.
K) the assay was accepted based on the measurement of cytotoxicity by relative total growth (RTG) which reduced to approximately 10-20% when compared to concurrent vehicle control. The mean vehicle control value for mutant frequency (MF) are between 50-170 x 10-6 The mean cloning efficiency is between 65-120%. The mean suspension growth are between 8-32 on kratom powder mixed with water day 2 (following 3 hr treatment with S9) After exclusion of obvious outliers at least 2 acceptable vehicle controls cultures remain.
The DNA profiles of three different cell lines (HEK 293 MCL-5 and SH-SY5Y cells) treated with MSE and MIT were assessed using nucleic acd staining with PI and analysed with BD FacsCalibur flow cytometer in the Centre for Molecular Microbiology and Infection (CMMI) core facility bali kratom and alcohol kratom legal in fl unit Flowers Building South Kensington Campus. The procedures were as described in section 4. Human embryo kidney- HEK 293 cells Using HEK 293 cells the effects of various concentration of MSE on the cell cycle profile was determined at 24 and 48 hr time period (Fig. The 10000 events were collected during the
acquisition and the phases of the cell cycle were gated manually using CellQuest Pro software.
Thus the combination consumption of Mitragyna speciosa Korth leaves with CYP 2E1 inducers may shift toxicity closer kratom effects strains to doses that are Purple Sticky Kratom 15x Extract 3 Grams pharmacologically active. Based on the current findings observed in the present maeng da kratom dosage capsule studies it is concluded that the methanol-chloroform extract (MSE) of the Mitragyna speciosa Korth (Kratom) leaves and its dominant alkaloid mitragynine (MIT) have potential to cause cytotoxicity to mammalian cells at high doses and is possibly harmful to human users. MIT is proposed to be a major contributor to MSE cytotoxicity. The main target system of MSE and MIT cytotoxicity is the central nervous system as shown by sensitivity of neuroblastoma cell lines (SH-SY5Y) throughout the studies.