Nitric Oxide Lab Report
Experiment: DNA damage and mutations in human cells when exposed to nitric oxide
Aim: To examine mutations after in vitro exposure of nitric oxide to human cells.
Abstract: Nitric oxide (NO) is a anatomical carrier formed by various cell types. In this experiment nitric oxide is made to react with undamaged human cells and solutions of DNA, RNA , guanine or adenine in aerobic conditions. TK6 human lymphoblastoid cells were altered. It has been observed that deamination of purines, pyrimidines and various forms of RNA , DNA under acidic conditions results in the formation of xanthine and hypoxanthine. Also presence of oxygen with low concentrations of nitric oxide in aqueous solutions may provoke nitrosyl donors. Nitric oxide in aerobic …show more content…
Cells in the medium were radio-labeled. Various concentrations(0.125, 0.25, 0.375, 0.5ml/ml) of nitric oxide were introduced for dose- response experiments and was incubated for 1 hour . Radioactivity was measured using scintillation counter.
Deamination of purines: Mild acid hydrolysis aided DNA purination. The mixture of water, HCl and xanthine in appropriate concentration were heated and an acid hydrolysate was formed which was anaylzed using HPLC(High Performance Liquid Chromatography) is based on the principle of seperation of components into parts as a result of the variation in their comparative affinities of individual molecules for the mobile phase and stationary phase of the separation. Results:
Nitric oxide cytotoxicity and mutagenecity: The addition of nitric oxide brought about a considerable concentration dependent decrease in survival and concentration dependent increase in mutant fraction in TK6 human lymphoblasts. Linear concentration dependency was seen from the graph. At highest nitric acid concentration(0.375ml) per ml of the culture medium showed increase in the amount of mutant fraction. The respective survival at higher concentration was found to be 0.12. All the concentrations of nitric oxide was found to be cytotoxic for the cells. A moderate decline in pH was observed due to the addition of nitric oxide. A pH range of pH 6.6-7.0 was seen at concentrations 0.125 and 0.25 repectively, of nitric oxide while nitric oxide concentration 0.375 showed pH of
Aim: To examine mutations after in vitro exposure of nitric oxide to human cells.
Abstract: Nitric oxide (NO) is a anatomical carrier formed by various cell types. In this experiment nitric oxide is made to react with undamaged human cells and solutions of DNA, RNA , guanine or adenine in aerobic conditions. TK6 human lymphoblastoid cells were altered. It has been observed that deamination of purines, pyrimidines and various forms of RNA , DNA under acidic conditions results in the formation of xanthine and hypoxanthine. Also presence of oxygen with low concentrations of nitric oxide in aqueous solutions may provoke nitrosyl donors. Nitric oxide in aerobic …show more content…
Cells in the medium were radio-labeled. Various concentrations(0.125, 0.25, 0.375, 0.5ml/ml) of nitric oxide were introduced for dose- response experiments and was incubated for 1 hour . Radioactivity was measured using scintillation counter.
Deamination of purines: Mild acid hydrolysis aided DNA purination. The mixture of water, HCl and xanthine in appropriate concentration were heated and an acid hydrolysate was formed which was anaylzed using HPLC(High Performance Liquid Chromatography) is based on the principle of seperation of components into parts as a result of the variation in their comparative affinities of individual molecules for the mobile phase and stationary phase of the separation. Results:
Nitric oxide cytotoxicity and mutagenecity: The addition of nitric oxide brought about a considerable concentration dependent decrease in survival and concentration dependent increase in mutant fraction in TK6 human lymphoblasts. Linear concentration dependency was seen from the graph. At highest nitric acid concentration(0.375ml) per ml of the culture medium showed increase in the amount of mutant fraction. The respective survival at higher concentration was found to be 0.12. All the concentrations of nitric oxide was found to be cytotoxic for the cells. A moderate decline in pH was observed due to the addition of nitric oxide. A pH range of pH 6.6-7.0 was seen at concentrations 0.125 and 0.25 repectively, of nitric oxide while nitric oxide concentration 0.375 showed pH of