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Title: The use and development of molecular biology techniques for human biomonitoring
Author: Stone, J. G.
Awarding Body: University College of Swansea
Current Institution: Swansea University
Date of Award: 1995
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The application of molecular techniques for the identification of cellular damage in response to environmental exposure is presented in this thesis. Using the 32P-postlabelling assay (32PPL) 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 10-7 in buccal mucosa and 1.66 X 10-7 in oral biopsies (p=0.72). Likewise no statistical difference between the two tissues was observed for smokers: 6.73 X 10-7 and 6.16 X 10-7 (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 P1 enhanced 32PPL 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 32PPL 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.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available