Browsing by Author "McCarthy, Keith"
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Item Open Access Detection of inappropriate cell proliferation in breast epithelium leading to breast cancer(2004-11) Degan, Simone A.; Woodman, Anthony C.; McCarthy, Keith; Bristol, James B.Breast cancer is predominantly caused by unrestrained cell proliferation. Proliferation is a complex process mediated by a network of signals that converge to a point called the ‘initiation of genome replication’ after which either proliferation or cell death could take place. The minichromosome maintenance (MCM) proteins are located at this point and play a pivotal role in regulating DNA replication. The detection of an aberrant level of such proteins can be of use in early breast cancer diagnosis. The main aim of this thesis was to propose a new system to detect inappropriate cell proliferation in breast epithelium. An in vitro model using cancer cell lines was developed to lay the foundation for subsequent studies employing human breast specimens. The application of the in vitro findings in breast excisions allowed assessing of the specificity and sensitivity of the biomarkers to ascertain slowly proliferating neoplastic cells. The most striking finding of this study was the abnormal presence of the MCM proteins in tumour compared to normal tissues with a typical pattern of expression unique for the histological classification of the lesion. The potential of MCM proteins as indicators of cell proliferation defects was further investigated with association studies with Ki-67, Bcl-2 and ER. MCM consistently identified a higher proportion of proliferating cells compared to Ki-67 suggesting that they are interesting markers of the Gi/S-phase. In fact, the MCM proteins start to co-localise in early Gi whereas Ki-67 is almost absent in this phase. Importantly, MCM proteins could recognise not only the proliferating compartment of the tumour but also those cells with replication potential. Based on these findings, the novel MCM biomarkers can be helpful in identifying both malignant and potentially malignant breast tissues. This feature can be useful in predicting patients at risk of tumour progression.Item Open Access Vibrational spectroscopy for the rapid and early diagnosis of leukaemias and lymphomas(Cranfield University, 2013-11) Jackson, Olivia; Stone, Nicholas; Bessant, Conrad; Rye, Adam; Lush, Richard; McCarthy, KeithThis thesis aimed to investigate vibrational spectroscopies for the identification of biochemical markers of leukaemias and lymphomas. In a preliminary study using the blood proteins albumin, fibrinogen and globulin, Drop Coating Deposition Raman Spectroscopy was explored and extended for use with Fourier Transform infrared spectroscopy for leukaemia blood sample analysis. Due to low sample volumes and minimal preparation required it was identified as a potential alternative to blood centrifugation to obtain the buffy coat for analysis. These studies identified that it was capable of detecting low levels of protein from small, highly concentrated droplets. Thus this method, alongside cytospin centrifugation, was used for the spectroscopic analysis of different blood fractions. Due to the low number of lymphoma samples obtained, only a feasibility study is outlined in this thesis. Samples were collected from leukaemia patients and healthy volunteers. Infrared and Raman spectra were measured of whole blood and buffy coat samples cytospun onto slides and whole blood and plasma pipetted by drop coating deposition. Multivariate statistical analysis was employed to extract key spectral differences between the pathologies and develop classification models for diagnosing chronic lymphoblastic leukaemia from previously treated and untreated patient groups. Principal component analysis followed by linear discriminant analysis was employed to identify the largest variances in the data and leave one sample out cross validation evaluated the performance of the spectral models measured on different blood components in diagnosing leukaemia. The buffy coat infrared model correctly classified 59% of the spectra, and blood droplet Raman 62%. The treated and untreated groups were then combined, which improved classification to 83% for buffy coat infrared and 71% for blood droplet Raman. These findings highlight the potential of drop coating deposition spectroscopy of whole blood for leukaemia diagnosis, although further work is required to achieve a clinically validated method.