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The application of mass spectrometry-based techniques to full thickness skin tissue: method development and biochemical analysis in health and disease

Bliss, EJ; (2016) The application of mass spectrometry-based techniques to full thickness skin tissue: method development and biochemical analysis in health and disease. Doctoral thesis , UCL (University College London). Green open access

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Abstract

The work presented in this thesis collates a selection of mass spectrometry-based techniques which have been applied to skin samples in order to identify biochemical changes expressed in the skin in health and disease. The results of an immunohistochemical study into the distribution of proteins within the skin revealed calmodulin-like protein 5 (CLP5) as having a different staining pattern between control and eczema samples. Further investigation revealed the role of CLP5 as a marker of keratinocyte differentiation and highlighted the importance of the calcium ion gradient in the skin for correct transport of proteases, protease inhibitors and antimicrobial proteins. Skin protease inhibitor and protease binding studies suggested that cystatin A could be a key player in manifestations of atopic eczema in susceptible children alongside two of its binding partners: dermcidin and caspase-14. Skin is a challenging tissue to analyse using traditional proteomic techniques due to the high lipid content, insolubility and extensive cross-linking of proteins. However, this thesis presents a mass spectrometry compatible method for its analysis and how that method has been applied to the study of hypertrophic scarring and post-operative morbidity. For hypertrophic scarring the most interesting and clinically significant perspective of this investigation was that there are changes in the endogenous profile of healthy skin that could predict whether a healthy scar or a hypertrophic scar will form after surgical injury. For the second clinical outcome (post-operative morbidity) we identified pre-operative hypoxia, antioxidant levels and reactive oxygen species (ROS) as indicators of post-operative morbidity. To conclude this thesis presents the work of a range of mass spectrometry techniques and has applied them to the study of human skin in health and disease. It demonstrates the versatility of mass spectrometry and has highlighted areas of clinical medicine where proteomics and a personalised medicine approach could be further investigated.

Type: Thesis (Doctoral)
Title: The application of mass spectrometry-based techniques to full thickness skin tissue: method development and biochemical analysis in health and disease
Event: UCL (University College London)
Open access status: An open access version is available from UCL Discovery
Language: English
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Pop Health Sciences > UCL GOS Institute of Child Health
URI: http://discovery.ucl.ac.uk/id/eprint/1528731
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