Optical and structural ocular changes during incipient presbyopia

The primary aim of this thesis was to investigate the in vivo ocular morphological and contractile changes occurring within the accommodative apparatus prior to the onset of presbyopia, with particular reference to ciliary muscle changes with age and the origin of a myopic shift in refraction during incipient presbyopia. Commissioned semi-automated software proved capable of extracting accurate and repeatable measurements from crystalline lens and ciliary muscle Anterior Segment Optical Coherence Tomography (AS-OCT) images and reduced the subjectivity of AS-OCT image analysis. AS-OCT was utilised to document longitudinal changes in ciliary muscle morphology within an incipient presbyopic population (n=51). A significant antero-inwards shift of ciliary muscle mass was observed after 2.5 years. Furthermore, in a subgroup study (n=20), an accommodative antero-inwards movement of ciliary muscle mass was evident. After 2.5 years, the centripetal response of the ciliary muscle significantly attenuated during accommodation, whereas the antero-posterior mobility of the ciliary muscle remained invariant. Additionally, longitudinal measurement of ocular biometry revealed a significant increase in crystalline lens thickness and a corresponding decrease in anterior chamber depth after 2.5 years (n=51). Lenticular changes appear to be determinant of changes in refraction during incipient presbyopia. During accommodation, a significant increase in crystalline lens thickness and axial length was observed, whereas anterior chamber depth decreased (n=20). The change in ocular biometry per dioptre of accommodation exerted remained invariant after 2.5 years. Cross-sectional ocular biometric data were collected to quantify accommodative axial length changes from early adulthood to advanced presbyopia (n=72). Accommodative axial length elongation significantly attenuated during presbyopia, which was consistent with a significant increase in ocular rigidity during presbyopia. The studies presented in this thesis support the Helmholtz theory of accommodation and despite the reduction in centripetal ciliary muscle contractile response with age, primarily implicate lenticular changes in the development of presbyopia.

Lichenometric dating (lichenometry) and the biology of the lichen genus rhizocarpon:challenges and future directions

Lichenometric dating (lichenometry) involves the use of lichen measurements to estimate the age of exposure of various substrata. Because of low radial growth rates and considerable longevity, species of the crustose lichen genus Rhizocarpon have been the most useful in lichenometry. The primary assumption of lichenometry is that colonization, growth and mortality of Rhizocarpon are similar on surfaces of known and unknown age so that the largest thalli present on the respective faces are of comparable age. This review describes the current state of knowledge regarding the biology of Rhizocarpon and considers two main questions: (1) to what extent does existing knowledge support this assumption; and (2) what further biological observations would be useful both to test its validity and to improve the accuracy of lichenometric dates? A review of the Rhizocarpon literature identified gaps in knowledge regarding early development, the growth rate/size curve, mortality, regeneration, competitive effects, colonization, and succession on rock surfaces. The data suggest that these processes may not be comparable on different rock surfaces, especially in regions where growth rates and thallus turnover are high. In addition, several variables could differ between rock surfaces and influence maximum thallus size, including rate and timing of colonization, radial growth rates, environmental differences, thallus fusion, allelopathy, thallus mortality, colonization and competition. Comparative measurements of these variables on surfaces of known and unknown age may help to determine whether the basic assumptions of lichenometry are valid. Ultimately, it may be possible to take these differences into account when interpreting estimated dates.

Improving T cell-induced response to subunit vaccines:opportunities for a proteomic systems approach

Prophylactic vaccines are an effective strategy to prevent development of many infectious diseases. With new and re-emerging infections posing increasing risks to food stocks and the health of the population in general, there is a need to improve the rationale of vaccine development. One key challenge lies in development of an effective T cell-induced response to subunit vaccines at specific sites and in different populations. Objectives: In this review, we consider how a proteomic systems-based approach can be used to identify putative novel vaccine targets, may be adopted to characterise subunit vaccines and adjuvants fully. Key findings: Despite the extensive potential for proteomics to aid our understanding of subunit vaccine nature, little work has been reported on identifying MHC 1-binding peptides for subunit vaccines generating T cell responses in the literature to date. Summary: In combination with predictive and structural biology approaches to mapping antigen presentation, proteomics offers a powerful and as yet un-tapped addition to the armoury of vaccine discovery to predict T-cell subset responses and improve vaccine design strategies.

The second intracellular loop of the calcitonin gene-related peptide receptor provides molecular determinants for signal transduction and cell surface expression

The calcitonin gene-related peptide (CGRP) receptor is a heterodimer of a family B G-protein-coupled receptor, calcitonin receptor-like receptor (CLR), and the accessory protein receptor activity modifying protein 1. It couples to Gs, but it is not known which intracellular loops mediate this. We have identified the boundaries of this loop based on the relative position and length of the juxtamembrane transmembrane regions 3 and 4. The loop has been analyzed by systematic mutagenesis of all residues to alanine, measuring cAMP accumulation, CGRP affinity, and receptor expression. Unlike rhodopsin, ICL2 of the CGRP receptor plays a part in the conformational switch after agonist interaction. His-216 and Lys-227 were essential for a functional CGRP-induced cAMP response. The effect of (H216A)CLR is due to a disruption to the cell surface transport or surface stability of the mutant receptor. In contrast, (K227A)CLR had wild-type expression and agonist affinity, suggesting a direct disruption to the downstream signal transduction mechanism of the CGRP receptor. Modeling suggests that the loop undergoes a significant shift in position during receptor activation, exposing a potential G-protein binding pocket. Lys-227 changes position to point into the pocket, potentially allowing it to interact with bound G-proteins. His-216 occupies a position similar to that of Tyr-136 in bovine rhodopsin, part of the DRY motif of the latter receptor. This is the first comprehensive analysis of an entire intracellular loop within the calcitonin family of G-protein-coupled receptor. These data help to define the structural and functional characteristics of the CGRP-receptor and of family B G-protein-coupled receptors in general. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.