apoptosis in cancer cells. The fruit rind of the plant Myristica malabarica, popularly known as rampatri, Bombay mace, or false nutmeg is used as an exotic spice in various Indian cuisines. Its pharmacological activities range from hepatoprotective, anti-ulcerogenic to anti-leishmanial. Its phytoconstituents include diarylnonanoids of which malabaricone-C showed potent anti-oxidant and anti-cancer activity which was attributed to its Cudependent nuclease property. In Leishmaniasis, a protozoan parasitic disease, the Leishmania parasites have an impaired anti-oxidant system wherein triggering of oxidative stress has been demonstrated to be an effective chemotherapeutic modality. Indeed, Miltefosine, that has anti-cancer and anti-leishmanial activity mediates its cytotoxicity via apoptosis. Therefore, considering that malabaricones have anti-leishmanial activity, it may be envisaged that it mediated its parasiticidal activity via its pro-oxidant property. Accordingly, we 1 MAL-A Causes ROS Induced Apoptosis tested the anti-cancer potential of malabaricones and whether their ability to achieve cell death was via redox imbalance. calculated by subtraction of background absorbance from total absorbance; the mean % viability was calculated as follows: Mean specific OD490 of treated cells 6100 Mean specific OD490 of untreated cells Each experiment was repeated thrice in duplicates and after the data was plotted, the IC50 i.e. the concentration that inhibited 50% cell growth was enumerated by graphical extrapolation using Graph Pad Prism software. PBMC was similarly incubated with MAL-A and cell viability measured as described above. Materials and Methods Materials All chemicals if not otherwise stated were obtained from SigmaAldrich except phenazine methosulphate, 5,5-dithiobis and trichloroacetic acid from PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22190027 Sisco Research Laboratories, MTS or 3–5–2–2H-tetrazolium, inner salt, 5, 59, 6, 69-tetrachloro-1, 19, 3, 39-tetraethylbenzimidazolylcarbocyanine iodide, fluo4 acetoxymethyl, 5-chloromethylfluorescein diacetate, 5–chloromethyl-29,79-dichlorodihydrofluorescein diacetate and Quick apoptotic DNA ladder detection kit, Z-Val-Ala-DL-Asp -fluoromethylketone, Caspase-3/CPP32, FLICE/caspase-8 and 193022-04-7 site caspase-9 colorimetric assay kit, Annexin-V FITC, Cell Death Detection Kit and 4,5 diaminofluorescein -2 diacetate, antibodies against poly polymerase and cytochrome c. Generation of intracellular reactive oxygen species and reactive nitrogen species CM-H2DCFDA, a lipid soluble membrane permeable dye upon entering cells undergoes deacetylation by intracellular esterases and forms the more hydrophilic, non-fluorescent dye Dichlorodihydrofluorescein. This is subsequently 
oxidized by ROS to form a two-electron highly fluorescent oxidation product, Dichlorofluorescein; therefore, the fluorescence generated is directly proportional to the amount of ROS. Similarly, DAF-2DA is freely permeable, undergoes hydrolysis by cytosolic esterases, resulting in the formation of DAF-2 which in the presence of intracellular NO gets converted into an impermeable, highly fluorescent triazolofluorescein. For both probes, fluorescence was acquired in a flow cytometer. The effect of a near IC50 concentration of MAL-A on generation of ROS and RNS was measured in Following centrifugation cell lines. and 2 washes, cells were resuspended in PBS, incubated for 30 minutes at 37uC with CM-H2DCFDA for measurement of ROS or DAF-2DA for RNS. Cell culture Three human cell lines namely U93
