Optimising the clinical analysis of retinal image quality in the human eye

Visual perception is dependent on both light transmission through the eye and neuronal conduction through the visual pathway. Advances in clinical diagnostics and treatment modalities over recent years have increased the opportunities to improve the optical path and retinal image quality. Higher order aberrations and retinal straylight are two major factors that influence light transmission through the eye and ultimately, visual outcome. Recent technological advancements have brought these important factors into the clinical domain, however the potential applications of these tools and considerations regarding interpretation of data are much underestimated. The purpose of this thesis was to validate and optimise wavefront analysers and a new clinical tool for the objective evaluation of intraocular scatter. The application of these methods in a clinical setting involving a range of conditions was also explored. The work was divided into two principal sections: 1. Wavefront Aberrometry: optimisation, validation and clinical application The main findings of this work were: • Observer manipulation of the aberrometer increases variability by a factor of 3. • Ocular misalignment can profoundly affect reliability, notably for off-axis aberrations. • Aberrations measured with wavefront analysers using different principles are not interchangeable, with poor relationships and significant differences between values. • Instrument myopia of around 0.30D is induced when performing wavefront analysis in non-cyclopleged eyes; values can be as high as 3D, being higher as the baseline level of myopia decreases. Associated accommodation changes may result in relevant changes to the aberration profile, particularly with respect to spherical aberration. • Young adult healthy Caucasian eyes have significantly more spherical aberration than Asian eyes when matched for age, gender, axial length and refractive error. Axial length is significantly correlated with most components of the aberration profile. 2. Intraocular light scatter: Evaluation of subjective measures and validation and application of a new objective method utilising clinically derived wavefront patterns. The main findings of this work were: • Subjective measures of clinical straylight are highly repeatable. Three measurements are suggested as the optimum number for increased reliability. • Significant differences in straylight values were found for contact lenses designed for contrast enhancement compared to clear lenses of the same design and material specifications. Specifically, grey/green tints induced significantly higher values of retinal straylight. • Wavefront patterns from a commercial Hartmann-Shack device can be used to obtain objective measures of scatter and are well correlated with subjective straylight values. • Perceived retinal stray light was similar in groups of patients implanted with monofocal and multi focal intraocular lenses. Correlation between objective and subjective measurements of scatter is poor, possibly due to different illumination conditions between the testing procedures, or a neural component which may alter with age. Careful acquisition results in highly reproducible in vivo measures of higher order aberrations; however, data from different devices are not interchangeable which brings the accuracy of measurement into question. Objective measures of intraocular straylight can be derived from clinical aberrometry and may be of great diagnostic and management importance in the future.

Novel glucose sensor based on enzymeimmobilized 81° tilted fiber grating

We demonstrate a novel glucose sensor based on an optical fiber grating with an excessively tilted index fringe structure and its surface modified by glucose oxidase (GOD). The aminopropyltriethoxysilane (APTES) was utilized as binding site for the subsequent GOD immobilization. Confocal microscopy and fluorescence microscope were used to provide the assessment of the effectiveness in modifying the fiber surface. The resonance wavelength of the sensor exhibited red-shift after the binding of the APTES and GOD to the fiber surface and also in the glucose detection process. The red-shift of the resonance wavelength showed a good linear response to the glucose concentration with a sensitivity of 0.298nm(mg/ml)-1 in the very low concentration range of 0.0∼3.0mg/ml. Compared to the previously reported glucose sensor based on the GOD-immobilized long period grating (LPG), the 81° tilted fiber grating (81°-TFG) based sensor has shown a lower thermal cross-talk effect, better linearity and higher Q-factor in sensing response. In addition, its sensitivity for glucose concentration can be further improved by increasing the grating length and/or choosing a higher-order cladding mode for detection. Potentially, the proposed techniques based on 81°-TFG can be developed as sensitive, label free and micro-structural sensors for applications in food safety, disease diagnosis, clinical analysis and environmental monitoring.

An introduction to analysis of variance (ANOVA) with special reference to data from clinical experiments in optometry

This article is aimed primarily at eye care practitioners who are undertaking advanced clinical research, and who wish to apply analysis of variance (ANOVA) to their data. ANOVA is a data analysis method of great utility and flexibility. This article describes why and how ANOVA was developed, the basic logic which underlies the method and the assumptions that the method makes for it to be validly applied to data from clinical experiments in optometry. The application of the method to the analysis of a simple data set is then described. In addition, the methods available for making planned comparisons between treatment means and for making post hoc tests are evaluated. The problem of determining the number of replicates or patients required in a given experimental situation is also discussed. Copyright (C) 2000 The College of Optometrists.

Analysis of clinical isolates of Propionibacterium acnes by optimised RAPD

Random amplification of polymorphic DNA (RAPD) was evaluated as a genotypic method for typing clinical strains of Propionibacterium acnes. RAPD can suffer from problems of reproducibility if parameters are not standardised. In this study the reaction conditions were optimised by adjusting template DNA concentration and buffer constituents. All isolates were typeable using the optimised RAPD protocol which was found to be highly discriminatory (Simpson's diversity index, 0.98) and reproducible. Typing of P. acnes by optimised RAPD is an invaluable tool for the epidemiological investigation of P. acnes for which no other widely accepted method currently exists. © 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Comprehensive human urine standards for comparability and standardization in clinical proteome analysis

Urine proteomics is emerging as a powerful tool for biomarker discovery. The purpose of this study is the development of a well-characterized "real life" sample that can be used as reference standard in urine clinical proteomics studies.

Analysis of secretory, immunostaining and clinical characteristics of human "functionless" pituitary adenomas:transdifferentiation or gonadotropinomas?

In this study, the central technique of in vitro culture has been used to further investigate whether LH/FSH-expressing, but clinically "functionless" pituitary adenomas are gonadotropinomas or whether their hormone secretion is due to transdifferentiation events. 664 "functionless" pituitary adenomas were examined for hormone secretion by in vitro culture and for hormone content by immunostaining. The results were correlated with the clinical findings. 40% of the tumours (n = 263) secreted at least one of the gonadotropins alone, 8% (n = 53) exhibited various patterns of anterior pituitary hormones, whilst the remaining 52% of tumours were not associated with any hormone. In the secretory tumours, immunostaining revealed only a few scattered hormone-containing cells (5 to 15%). Mild hyperprolactinaemia was observed in some cases, presumably because of pressure effects of the tumours. The majority of the patients suffered clear cut hypopituitarism (p < 0.05). Pre-operatively, gonadotropin hypersecretion was observed in 3 cases, but only one of these secreted hormones in culture. Interestingly, a higher proportion of tumours removed from patients with hypopituitarism showed secretory activity in vitro than those tumours removed from patients showing no hormonal dysfunction or hyperprolactinaemia. We conclude that the term "gonadotropinoma" to describe functionless pituitary tumours associated with LH and/or FSH secretion is a misnomer, because the presence of LH and/or FSH confirmed by in vitro methods in the present series is a result of only a few scattered cells. We suggest that primary pituitary tumour cells differentiate into a secretory type (transdifferentiation), possibly in response to altered serum hormone levels such as decreased steroids. Further work is required to identify the factors which trigger the altered cells' characteristics. © J. A. Barth Verlag in Georg Thieme Verlag KG.

Pronounced reduction of postprandial glucagon by lixisenatide:a meta-analysis of randomized clinical trials

Glucagon-like peptide-1 (GLP-1) receptor agonists improve islet function and delay gastric emptying in patients with type 2 diabetes mellitus (T2DM). This meta-analysis aimed to investigate the effects of the once-daily prandial GLP-1 receptor agonist lixisenatide on postprandial plasma glucose (PPG), glucagon and insulin levels. Methods: Six randomized, placebo-controlled studies of lixisenatide 20μg once daily were included in this analysis: lixisenatide as monotherapy (GetGoal-Mono), as add-on to oral antidiabetic drugs (OADs; GetGoal-M, GetGoal-S) or in combination with basal insulin (GetGoal-L, GetGoal-Duo-1 and GetGoal-L-Asia). Change in 2-h PPG and glucose excursion were evaluated across six studies. Change in 2-h glucagon and postprandial insulin were evaluated across two studies. A meta-analysis was performed on least square (LS) mean estimates obtained from analysis of covariance (ANCOVA)-based linear regression. Results: Lixisenatide significantly reduced 2-h PPG from baseline (LS mean difference vs. placebo: -4.9mmol/l, p<0.001) and glucose excursion (LS mean difference vs. placebo: -4.5mmol/l, p<0.001). As measured in two studies, lixisenatide also reduced postprandial glucagon (LS mean difference vs. placebo: -19.0ng/l, p<0.001) and insulin (LS mean difference vs. placebo: -64.8 pmol/l, p<0.001). There was a stronger correlation between 2-h postprandial glucagon and 2-h PPG with lixisenatide than with placebo. Conclusions: Lixisenatide significantly reduced 2-h PPG and glucose excursion together with a marked reduction in postprandial glucagon and insulin; thus, lixisenatide appears to have biological effects on blood glucose that are independent of increased insulin secretion. These effects may be, in part, attributed to reduced glucagon secretion. © 2014 John Wiley and Sons Ltd.

Clinical monitoring of ocular physiology using digital image analysis

Aim: To examine the use of image analysis to quantify changes in ocular physiology. Method: A purpose designed computer program was written to objectively quantify bulbar hyperaemia, tarsal redness, corneal staining and tarsal staining. Thresholding, colour extraction and edge detection paradigms were investigated. The repeatability (stability) of each technique to changes in image luminance was assessed. A clinical pictorial grading scale was analysed to examine the repeatability and validity of the chosen image analysis technique. Results: Edge detection using a 3 × 3 kernel was found to be the most stable to changes in image luminance (2.6% over a +60 to -90% luminance range) and correlated well with the CCLRU scale images of bulbar hyperaemia (r = 0.96), corneal staining (r = 0.85) and the staining of palpebral roughness (r = 0.96). Extraction of the red colour plane demonstrated the best correlation-sensitivity combination for palpebral hyperaemia (r = 0.96). Repeatability variability was <0.5%. Conclusions: Digital imaging, in conjunction with computerised image analysis, allows objective, clinically valid and repeatable quantification of ocular features. It offers the possibility of improved diagnosis and monitoring of changes in ocular physiology in clinical practice. © 2003 British Contact Lens Association. Published by Elsevier Science Ltd. All rights reserved.