Thesis (Ph. D.)--Brandeis University, 2010. / "UMI:3390495." MICROFILM COPY ALSO AVAILABLE IN THE UNIVERSITY ARCHIVES. Includes bibliographical references.

No description available.

No description available.

No description available.

Problems with the low back resulting from diseases of the lumbar disc account for much human suffering and medical expense. Through early identification of changes in the disc, Magnetic Resonance Imaging offers a potential method for recognition of those at risk of low back trouble. An understanding of the chemical and structural basis of the MR image will therefore be of great help in understanding the degenerative mechanism itself. With this in mind, NMR studies of human intervertebral discs were performed to extract data that could be correlated with different stages of degeneration. In vitro samples of anulus fibrosus and nucleus pulposus of grades II and IV discs were examined. The Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence was used to obtain T₂ relaxation data, which were analysed with a non-negative least squares algorithm and presented as a continuous spectrum of exponentials. An inversion-recovery sequence was used to obtain T₁ data which were analysed with the same algorithm, then presented as a discrete exponential spectrum. The same procedures were used to produce relaxation times from samples of isolated disc collagen and proteoglycan. Also, T₂ relaxation data were obtained with a twelve-echo CPMG imaging sequence from healthy volunteers, and these data analysed with the same procedure used for in vitro data. In vivo and in vitro results were compared and found to agree in terms of the T₂ relaxation values. Most change between grades occurred in the nucleus, where both T₁ and T₂ values decreased from grade II to grade IV. The results of the proteoglycan and collagen studies support the theory that structure, not chemical composition, is responsible for the observed changes in relaxation times. Based on estimates of the percentages of protons in the nucleus due to water, proteoglycan and collagen, tentative assignments were made to the components of the T₂ spectra for grades II and IV nucleus, and these were supported by the results of all parts of the study. / Science, Faculty of / Physics and Astronomy, Department of / Graduate

The pathogenesis of low back pain is complex but likely involves the intervertebral disc (Nachemson, 1976). Direct evidence for its importance is lacking because an accurate in vivo method of imaging the lumbar intervertebral disc has not been established. The objective of this research was to develop classifications of gross morphologic appearance and magnetic resonance image (MRI) of the disc that describe the changes associated with aging and degeneration, thereby permitting interpretation of the MRI in terms of gross morphology and allowing correlation of morphologic, chemical, mechanical, radiologic and epidemiologic data with a standard reference of disc aging and degeneration. The classifications were developed on the basis of literature review, detailed examination of 55 discs and expert advice. Two sets of three observers, one for the morphologic classification and one for the MRI classification evaluated 68 life size randomized duplicates of discs making detailed observations about overall category and 17 regional morphologic parameters and 11 regional MRI parameters. The data was tested to demonstrate the validity of the classifications using established criteria (Tugwell & Bombardier, 1982; Guyatt 4 Kirschner, 1985; Feinstein, 1985). The consistency with which the classifications could be applied was evaluated by calculating weighted kappa, a statistical test of agreement that corrects for agreement by chance; the ability of the classifications to distinguish stages in the process of ageing and degeneration by stepwise discriminant analysis; their conformity with other measures by comparisons within and between classifications and, comparisons with histologic and chemical data. The degree of agreement for all six intra-observer pairs was 'almost perfect' (weighted kappa > 0.80); for 5 interobserver pairs 'substantial' (weighted kappa > 0.60) and for one MRI interobserver pair 'moderate' (weighted kappa > 0.50). This represented a satisfactory level of agreement and indicated the classifications could be applied consistently (Feinstein, 1981). The linear regression model developed by stepwise discriminant analysis clearly demonstrated the ability of the classifications to distinguish distinct stages in disc aging and degeneration. Wilk's lambda, a likelihood ratio statistic reflecting discriminatory function, approached zero in both the morphologic (0.0408) and MRI (0.0H80) classifications. In both models, parameters pertaining to the nucleus pulposus of the disc accounted for the majority of the variance (morphologic partial R² 0.8598 and MRI partial R² 0.8811) suggesting nuclear parameters are the most important in distinguishing overall category. The correlation table generated by principal component analysis demonstrated that the categories assigned to regional parameters correlated significantly (p > 0.0001) with each other and with the overall category. From the linear combinations of parameters (principal components) generated the weighting of the nucleus pulposus behaved independently attesting to its importance. Comparisons of the morphologic and MRI classifications yielded high indices of trend (Pearson correlation coefficient of 0.81) and concordance (kappa of 0.62). Trends in the histologic and chemical data were consistent with the classifications but could not be evaluated statistically because only 15 specimens were studied. This research suggests that the classifications are valid and will form a basis for the interpretation of MRI. Preliminary evidence suggested MRI is sensitive to early changes in extracellular matrix composition not apparent in gross morphology. / Medicine, Faculty of / Graduate

In the present work three different proteins were investigated, namely IGPS, the SRII/HtrII complex and ChR2. EPR spectroscopy combined with SDSL were applied to investigate the relation of the respective protein dynamics and their functions. In case of IGPS spin label side chain dynamics were recorded and allow a classification of loop dynamics during catalysis. For SRII/HtrII an equilibrium of spin label side chain dynamics was found to reflect the equilibrium of the activated and deactivated states, which are responsible for signal transfer through the complex. Time-resolved analyses of side chain dynamics and trapping of intermediates of the light-triggered channel ChR2 identified conformational changes to occur during channel function. Thereby, the EPR spectroscopy proves to be a powerful tool to investigate proteins in context of structural changes during their protein function.

No description available.

No description available.

No description available.