For cytological examinations Rapi-Diff staining was purchased from Bios Europe U. Wright-Giemsa staining was from Sigma-Aldrich U. The opioid receptor antagonists naloxone naltrindole and cyprodime hydrobromide were purchased from Sigma-Aldrich U. Kratom Whole Leaf Milo iV set was from Calbiochem U. Caspase -8 and Caspase-9 Protease Kits were from Invitrogen U. The fluorescent dye 27-dichlorofluorescein diacetate Kratom Whole Leaf Milo (DCFH-DA) and hydrogen peroxide (H202) for ROS assay were purchased from Sigma-Aldrich U.
Prior to this study most of the investigations on the biological effects of this plant such as antinociceptives effects were mostly comparisons with opiate drugs such as morphine and its related compounds. Thus an important issue is whether MSE or MIT induced cell death may share similar mechanisms as opiate induced cell death. In general opioids have been shown to induce in vitro apoptosis in cell lines including neuronal cells (Mao et al 2002). The recent review by Zhang et al (2008) stated that morphine for instance induces neurotoxicity and apoptosis after chronic use and heroin also induced apoptotic ultimate thai kratom cell death via mitochondrial malfunction caspase activation leading to PARP cleavage and DNA fragmentation. Thus MIT may show a similar trend of apoptotic cell death as opiates but confirmation of this finding requires further investigations. MSE as death appears to be caspase-independent and thus chemicals other than MIT present in MSE appear to complicate the interpretation of my biochemical findings. Despite having a crucial role for cellular energy metabolism mitochondria are also known to be a key player in cell death.
Wound- healing assay. Properties of purified liver microsomal cytochrome P450 from ralts treated with the polychlorinated biphenyl mixture arochlor 1254. Molecular Pharmacology 13: 521-532. Programmed cell death in development.
Although to date there is no report of cancer associated with consuming the leaves of this plant a genotoxic assessment such as mutagenicity aids prediction of carcinogenicity potential. Thus for the first time I have shown that genotoxicity testing using the mouse lymphoma tk gene mutation assay (MLA) suggests that MSE and MIT have no genotoxic potential. This MSE toxicity was similar to that noted for MSE with the human cell lines (SH-SY5Y and HEK 293 cells) in the presence of S9.
The Medical Journal of Australia 166:538-541. CIP1 is induced in p53-mediated G1 arrest and apoptosis. WAF1 a potential of p53 tumor suppression. Cell 75: 817-825. kratom liquid erowid trail Measuring mitochondrial reactive oxygen species. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and kratom withdrawal anxiety removal by macrophages. Preface: Cannabinoids as new tools for the treatment of neurological disorders.
In order to assess these effects more fully the well established Modfit software was employed for more detailed cell cycle analysis. In general the DNA profiles for MSE treated MCL-5 cells (Fig. MSE table 2.
Hypotheses 57: 96-100. Killing tumours by ceramide-induced apoptosis: a critique of available drugs. Double identity for protein of the Bcl-2 family. Nature 387: 773-776.
To further examine the involvement of metabolism in MSE and MIT associated toxicity specific inhibitors of metabolic enzymes were used. M ketoconazole (KT) a CYP 3A4 inhibitor (Gibbs et al. M 3-amino-124-triazole (ATZ) a CYP2E1 inhibitor (Koop 1990). C in 5% CO2). AbD Serotec U.
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Effect of MIT on cell cycle distribution of SH-SY5Y cells after 24 hr treatment. Histograms and values of the cell cycle phases are representative of a single experiment analysed by Modfit software. Protein concentrations of the cell lysates The bicinchoninic assay (BCA) is quick and works in a similar way to the Lowry method.
However due to its narcotism properties it has been misused by drug addicts as an alternative to opium or to moderate the withdrawal symptoms of opium. After years of research with this plant mainly using crude alkaloid extracts its dominant alkaloid mitragynine (MIT) and congeners their analgesic properties have been confirmed in vitro and in vivo. This medicinal property has so far been reported in the leaves 15x kratom extract of this plant but not from other species of
Mitragyna. Several countries like Thailand Myammar Malaysia and recently Australia have made this plant illegal due to its narcotism properties whereas in other parts of the world the plant regardless of any form has been sold widely over the internet.
Models of reactive oxygen species in cancer. Drug Discov Today Dis Models 4: 67-73. F Lai C.
The basis of the assay is measurement of fluorescence due to the release of lactate dehydrogenase (LDH) from cells with a damaged membrane. After 24 hr of treatment there was a dose-dependant toxicity trend seen with the MSE (Fig. However the trend towards toxicity was only seen at doses of MSE in excess of 0.
This can be stored for later use. Small pellets of this extract (which is also sold as such in various shops) can be swallowed or can be dissolved in hot water and consumed as a tea. Some people like to mix kratom tea with ordinary black tea or other herbal teas before it is consumed. Sugar or honey can be added to sweeten it.