The application of molecular techniques for the identification of cellular damage in response to environmental exposure is presented in this thesis. Using the <SUP>32</SUP>P-postlabelling assay (<SUP>32</SUP>PPL) buccal mucosa has been shown to represent an easily obtainable, alternative tissue for human biomonitoring. Relative adduct levels (RAL) in oral biopsies and buccal mucosa, taken from smokers and non-smokers, were analysed. Mean RAL for non-smokers were not statistically different in the two tissues: 2.1 X 10<SUP>-7</SUP> in buccal mucosa and 1.66 X 10<SUP>-7</SUP> in oral biopsies (p=0.72). Likewise no statistical difference between the two tissues was observed for smokers: 6.73 X 10<SUP>-7</SUP> and 6.16 X 10<SUP>-7</SUP> (p=0.75) respectively. Statistically higher levels of damage were seen in smokers compared to non-smokers. The p53 mutation spectra of oral tumours, obtained from smokers and non-smokers, was assessed using PCR amplification and Sanger sequencing. Four mutations were found: two in exon 7 at codon 247 (AAC to AC); one in exon 8 at codon 294 (GAG to GAAG) and one in codon 299 (CTG CCC to CTG AA CCC). No relationship was found between cigarette smoking and p53 mutation. A small study was carried out to assess persons living in an industrial area of South Wales. Using the butanol and nuclease P<SUB>1</SUB> enhanced <SUP>32</SUP>PPL methods to measure DNA adducts in WBC revealed the mean RAL to be statistically higher in the exposed group than the control group (p=<0.05). Analysis of buccal mucosa samples by butanol <SUP>32</SUP>PPL revealed no statistical difference between RAL in the exposed and control groups (p=>0.05). The p53 gene in exposed persons was analysed for mutations. WBC were analysed using the Restriction Site Mutation assay. No mutations were found.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:639116 |
| Date | January 1995 |
| Creators | Stone, J. G. |
| Publisher | Swansea University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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