The nature of cell death and mechanism associated with it is yet to be reported. Thus in this part of this thesis several investigations were attempted to provide possible mechanism of the nature and mode of cell death seen with a selected panel of human cell lines. The cytological examination using three different cell lines (SH-SY5Y HEK 293 and MCL-5 cells) was the first investigation.
The recent review by Zhang et al (2008) stated best opiate in renal failure that morphine for instance induces neurotoxicity and apoptosis after chronic use and heroin also induced apoptotic cell death via mitochondrial malfunction caspase activation leading to PARP cleavage and DNA fragmentation. Mitragyna Speciosa Half Life Kewaskum thus MIT may show a similar trend of apoptotic cell death as opiates but confirmation of this finding requires mitragyna speciosa also known as kratom further investigations. MSE as death appears to be caspase-independent and best opiate detox method thus chemicals other than MIT present in MSE appear to complicate the interpretation of my biochemical findings.
Carcinogens are mutagens: A simple test system combining liver homogenates for activation and bacteria for detection. PNAS 70: 2281-2285. Conjugation-dependant carcinogenicity and toxicity of foreign compounds Advances in Pharmacology 27: 1-512.
B MIT Treatment without S9 (24 hr) Neg. C 30 20 10 5 MMS Cell conc. Relative suspension growth (RSG) 100.
Mutant frequency was determined by seeding a known number of cells in medium containing TFT to detect mutant cells and also in medium without TFT to determine the cloning efficiency Mitragyna Speciosa Half Life Kewaskum (viability). Colonies were counted after 7 days for viability. The mutant frequency was determined after 11 days incubation and the size of colonies was assessed Mitragyna Speciosa Half Life Kewaskum according to the criteria described in section 3.
The mechanism for cell cycle arrest in the cells treated with high doses of MSE remains unclear as there was no correlation with p53 and p21 as both proteins were lost after the treatment. The level of MSE toxicity for SH-SY5Y and HEK 293 cells was found to be increased 10-fold when metabolic activation system (post mitochondrial rat liver S9 induced with Arochlor 1254) was added to the treatment. This implies that MSE cytotoxicity requires metabolism for its activation and CYP2E1 was thought to be involved in this metabolic activation. However MIT in parallel experiments did not show any enhancement of toxicity in the presence of S9 and was inherently cytotoxic.
C and D). At the 24
hr time point of both caspase assays (Fig. Activity of initiator caspase 8 after A) thai kratom nausea 4 hr incubation and B) 24 hr incubation time period and initiator caspase 9 after C) 4 hr incubation and D) 24 hr incubation time period of SH-SY5Y cells treated with MSE. The reading of each concentration is from 2 pooled lysates.
Darynkiewicz et al 2001; van Engeland et al 1998). C (5% CO2) for 24 hour. After routine harvesting as described in chapter 2 section 2. PBS followed by centrifugation (1200 r.