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Created with Fabric.js 1.4.5 ANGLO-CHINESE SCHOOL (INDEPENDENT)LIM JIA YI JOEL ME 044 METHOD & MATERIALS: 97.2% HYPOTHESIS 29.190 7.7% Ischemic brain injuryPro-apoptotic properties ISI O2 --> O2- --> H2O2 --> OH Oxidative stress To investigate the effect of Amaranthus viridis leaf polyphenols on cell aging due to hydrogen peroxide and other mitochondria-generated oxidants Polyphenols Antioxidant capacity Mitochondria Aerobic Respiration 97.2% 1 % survival of baker's yeast colonies Aging, Cancer, CataractsAlzheimer's Disease Parkinson's Disease Increase in extract concentration --> increase in in vitro antioxidant capacity --> reduces replicative aging--> more daughter cells produced --> increase in area of CFU --> increase in % survival of cells --> no significant difference for freshly-harvested & 7 days post-harvest leaves (t- test) 4 3 Growing of plants - Closed static hydroponics system - Leaves harvested at Week 15 - Set 2: Stored at 4 C for 7 days Extraction of polyphenols - 95% ethanol in Moulinex electric blender (high power, 60.00 s) - Filter with filter funnel and paper - Evaporate ethanol using Eyela rotary evaporator (37 C, 100 rpm) - Redissolve in 100 cm3 (±1) of deionized water --> 200, 400, 600, 800, 1000 g dm-3 Preparation of baker's yeast cells - 10.0 cm3 (±0.1) of nutrient broth - 10.0 cm3 (±0.1) of extract - 78.2 ul (±0.1) of 6% H2O2 - Incubate for 5 h (32.0 C) Replication of baker's yeast cells - Sarourand Dextrose Agar in 100 mm petri dish - T-streak four quadrant method - Incubate for 48 h (32.0 C) Determination of in vivo antioxidant capacity - Area of baker's yeast colony forming units (CFU) in fourth quadrant - % area of baker's yeast CFU - % of survival of baker's yeast colonies 2 - Absorption, first-pass metabolism: Caco-2 human intestinal epithelium- More prolonged post-harvest storage - Polyphenol toxicity 5 FUTURE WORK: Increasing the amount of polyphenols in Amaranthus viridis (Green Amaranth) leaves, freshly-harvested and 7 days post-harvest, increases its in vivo antioxidant capacity interms of percentage survival of Saccharomyces cerevisiae (bakers yeast) colonies as higher concentrations of extract will have greater amounts of polyphenols with more phenolic hydroxyl groups to scavenge the OH generated by the added H2O2. Model for aging of mitotically-active cell types eg. haemopoetic stem cells Replicative lifespan: No. of daughtercells produced before senescence Replicative Aging RESULTS: CONCLUSION: Image of structure of quercetin taken from wikipedia commonsOther images by candidate Phenolic compounds (POH) donate H atom to form relatively stablephenoxy radical intermediates (PO) POH + OH --> H2O + POPO + OH --> POOH
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