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Title: Novel strategies for the detection of ragwort poisoning in horses
Author: Moore, Rowan Elizabeth
ISNI:       0000 0004 2672 6125
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2008
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Toxic pyrrolizidine alkaloids (PAs) are produced by flowering plant species throughout the world, including ragwort (Senecio jacobea). In animals, PAs are metabolised within the liver to form their corresponding dehydro-PAs or pyrroles. These compounds are highly reactive and are capable of alkylating and cross-linking DNA and proteins to lead to characteristic molecular and cellular insult within the liver. There are presently no metllOds for tlle definitive diagnosis of ragwort poisoning in animals. If biomarkers for ragwort poisoning could be identified in dIe early stages of ingestion dlen the onset of irreversible patllology might be preventable. The goal of this study was to explore methods for the detection and analysis of pyrrole-peptide and pyrrole-protein adducts· in vitro and to investigate techniques facilitating the identification of changes in equine blood as a result of PA poisoning in Vivo. Initial experiments aimed to elucida~e dIe range of responses of model proteins to alh.'ylating agents ill tll/ro. Intact mass analysis of dIe proteins by l-dimensional gel electrophoresis (lD-GE) and quadtupole-time-of-flight mass spectrometry (Q-ToF MS) demonstrated that different reagents are capable of distinctly modifying proteins, and that these reagents have varying degrees of specificity for dIe sulphydryl group of cysteine residues. Matri.x-assisted laser desorption/ionisation (MALDI)-ToF MS revealed the particular regions of proteins which were targeted by reagents and demonstrated that peptide adduct detection could be enhanced by guanidination of lysine-terminated peptides.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available