Doug Coster

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Apoptosis is a prominent feature of acute anterior uveitis in the Fischer 344 rat
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2000-02) Smith, Justine R; Hart, Prue; Standfield, Scott D; Coster, Douglas John; Wing, Sarah J; Williams, Keryn Anne
AIMS: To examine the hypothesis that apoptosis of infiltrating cells contributes to spontaneous resolution of uveitis in clinically relevant rodent models. METHODS: Experimental melanin induced uveitis (EMIU) was induced in Fischer 344 rats by immunisation with 250 microg bovine ocular melanin. Endotoxin induced uveitis (EIU) was induced by injection of 200 microg Escherichia coli lipopolysaccharide. Formalin fixed, paraffin embedded ocular cross sections were stained by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate biotin nick end labelling (TUNEL) to identify apoptotic cells. Indirect immunoperoxidase staining of paraformaldehyde lysine periodate fixed tissue cross sections was used to demonstrate expression of inducible nitric oxide synthase (iNOS). RESULTS: TUNEL positive mononuclear cells were observed in the anterior uvea during both EMIU and EIU at all selected time points. However, whereas the majority of mononuclear cells appeared apoptotic from the outset of disease, neutrophils were notably TUNEL negative at all time points examined. Many infiltrating neutrophils expressed iNOS. CONCLUSION: Apoptosis occurs early in the course of rat EMIU and EIU, and may contribute to resolution of these diseases. In general, infiltrating mononuclear cells die rapidly, while neutrophils survive, producing inducible nitric oxide synthase which may contribute to disease pathogenesis.
The Australian Corneal Graft Registry 2004 annual report
(2006-06-20T03:38:28Z) Williams, Keryn Anne; Hornsby, Ngaere B; Bartlett, Christine Mary; Holland, Helene K; Esterman, Adrian Jeffrey; Coster, Douglas John
The Australian Corneal Graft Registry (ACGR) opened in May 1985 and has now been in operation for 18 years. In that time, we have collected data on over 14,000 corneal grafts. At the time of transplantation, we seek information on the recipient, the donor and the operative procedure. Follow-up then occurs at approximately yearly intervals for an indefinite period. Follow-up only ceases upon loss of the graft, or death or loss-to-follow-up of the patient. At each round of follow-up, we request information on graft and visual outcome, and upon relevant events and treatments. The data are entered into a database and checked for consistency. Descriptive, univariate and multivariate analyses are performed, and the report collated.
The Australian Corneal Graft Registry 2007 Report
(Flinders University Press, 2007) Williams, Keryn Anne; Lowe, Marie Therese; Bartlett, Christine Mary; Kelly, L; Coster, Douglas John
The Australian Corneal Graft Registry opened in May 1985 and thus has now been in operation for over 22 years. The census date for this report was 01/09/2006. Over the years, we have collected data on more than 18,500 corneal grafts. The majority of corneal grafts registered have been penetrating, but increasing numbers of lamellar and limbal grafts have also been registered over recent years, as patterns of surgical practice change. At registration, we seek information on the recipient, the donor, the eye bank practices and the operative procedure. Follow-up then occurs at approximately yearly intervals for an indefinite period, and ceases upon loss of the graft, or the death or loss-to-follow-up of the patient. At each round of follow-up, we request information on the graft and visual outcome, and upon relevant post-operative events and treatments. The data are entered into an Access database and checked for consistency. Descriptive, univariate and multivariate analyses are subsequently performed using SPSS and Stata software, and the report is eventually collated.
The Australian Corneal Graft Registry 2012 Report
(2012-03-09) Williams, Keryn Anne; Lowe, Marie Therese; Keane, Miriam Claire; Jones, Victoria J; Loh, Raymond S K; Coster, Douglas John
The Australian Corneal Graft Registry (ACGR) opened in May 1985 and thus has now been in operation for over 26 years. However, the census dates for this report was 01/06/2010 for penetrating grafts and 12/10/2011 for lamellar grafts. Over the years, we have collected data on more than 23,000 corneal grafts. The majority of corneal grafts registered have been penetrating, but increasing numbers of lamellar grafts have also been registered over recent years, as patterns of surgical practice change. At registration, we seek information on the recipient, the donor, the eye bank practices and the operative procedure. Follow-up then occurs at approximately yearly intervals for an indefinite period, and ceases upon loss of the graft, or the death or loss-to-follow-up of the patient. At each round of follow-up, we request information on the graft and visual outcome, and upon relevant post-operative events and treatments. The data are entered into an Access database and checked for consistency. Descriptive, univariate and multivariate analyses are subsequently performed using SPSS and Stata software, and the report is eventually collated.
The Australian Corneal Graft Registry 2015 Report
(South Australian Health and Medical Research Institute, 2015-06-15) Williams, Keryn Anne; Keane, Miriam Claire; Galettis, Rachel A; Jones, Victoria J; Mills, Richard Arthur; Coster, Douglas John
The Australian Corneal Graft Registry (ACGR) opened in May 1985 and has now been operating for 30 years. Over the years, we have collected information on more than 30,000 corneal grafts. At registration, we seek information on the donor, eye bank practices, the recipient, the surgeon, the graft type and the operative procedure. Follow-up then occurs at approximately yearly intervals for an indefinite period, and ceases upon graft failure, or the death or loss-to-follow-up of the patient. At each round of follow-up, we request information on the survival of the graft, the visual outcomes, and any relevant post-operative events and treatments. The data are entered into an Access database and checked for consistency. Descriptive, univariate and multivariate analyses are subsequently performed using SPSS and Stata software, and the report is eventually collated.
Contrast and glare testing in keratoconus and after penetratin keratoplasty
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2004-05) Pesudovs, Konrad; Schoneveld, P; Seto, R J; Coster, Douglas John
AIM: To compare the performance of keratoconus, penetrating keratoplasty (PK), and control subjects on clinical tests of contrast and glare vision, to determine whether differences in vision were independent of visual acuity (VA), and thereby establish which vision tests are the most useful for outcome studies of PK for keratoconus. METHODS: All PK subjects had keratoconus before grafting and no subjects had any other eye disease. The keratoconus (n = 11, age 35.0 (SD 11.1) years), forme fruste keratoconus (n = 6, 33.0 (13.0)), PK (n = 21, 41.2 (7.9)), and control (n = 24, 33.7 (8.6)) groups were similar in age. Vision testing, conducted with optimal refractive correction in place, included low contrast visual acuity (LCVA) and Pelli-Robson contrast sensitivity (PRCS) both with and without glare, as well as VA. RESULTS: Normal subjects saw better than PK subjects who in turn saw better than keratoconus subjects on all raw measures. However, when adjusted for VA, the normal group only saw significantly better than the keratoconus group on LCVA (low contrast loss 0.05 (0.04) v 0.15 (0.12), F(2,48) = 6.16; p<0.01, post hoc Sheffe p<0.05), and the decrements to glare were no worse than for normals. The forme fruste keratoconus group were indistinguishable from normals on all measures. CONCLUSIONS: PK subjects have superior vision to keratoconus subjects, but not as good as normal subjects. Including mild keratoconus subjects within a keratoconus group could confound these differences in vision. While VA is an excellent test for comparing normal, keratoconus and PK groups, additional information can be provided by LCVA and PRCS, but not by glare testing. Outcomes research into keratoconus management should include a measure in the contrast domain.
Corneal graft rejection occurs despite Fas ligand expression and apoptosis of infiltrating cells
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2005-05) Williams, Keryn Anne; Standfield, Scott D; Smith, Justine R; Coster, Douglas John
BACKGROUND/AIMS: Constitutive expression of Fas ligand (CD95L) protects the eye against cell mediated immune responses by inducing apoptosis in infiltrating Fas bearing T cells. This study was designed to examine Fas ligand expression on acutely rejecting rat corneal grafts and to investigate the kinetics of induction of apoptosis in infiltrating leucocytes. METHODS: Orthotopic penetrating corneal transplantation was performed between genetically disparate inbred rats. Fas ligand expression and the phenotype of infiltrating leucocytes were examined by immunohistochemistry. Apoptotic nuclei were visualised in sections of normal rat cornea, rejecting allografts, and time matched isografts by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labelling (TUNEL) and quantified by video image analysis. Staining with Hoechst dye 33258 was used to confirm the presence of apoptotic nuclei. RESULTS: Fas ligand was expressed on corneal endothelial and epithelial cells during acute corneal graft rejection. At all time points examined, including as early as the fifth postoperative day, the cells infiltrating both corneal isografts and allografts were TUNEL positive. By the 15th postoperative day, over 90% of all nuclei, many of which were T cells, were apoptotic. CONCLUSION: Expression of Fas ligand is not downregulated on the cornea during allograft rejection and infiltrating leucocytes in both isografts and allografts die rapidly in situ. Despite the death of the cells believed to be responsible for rejection, isografts survive indefinitely whereas allografts are irreparably damaged.
Evaluation of corneal transplantation
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 1997-08) Coster, Douglas John
Editorial / comment on corneal transplantation.
In vitro adenovirus mediated gene transfer to the human cornea
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2005-06) Jessup, Claire Frances; Brereton, Helen Mary; Coster, Douglas John; Williams, Keryn Anne
BACKGROUND/AIMS: Replication deficient adenovirus is an efficient vector for gene transfer to the cornea. The aim was to optimise the transduction of human corneal endothelium with adenoviral vectors and to measure transgene production from transduced corneas. METHODS: Adenoviral vectors (AdV) encoding enhanced green fluorescent protein (eGFP) or a transgenic protein (scFv) were used to transfect 34 human corneas. Reporter gene expression was assessed after 72-96 hours of organ culture. The kinetics of scFv production was monitored in vitro for 1 month by flow cytometric analysis of corneal supernatants. RESULTS: Transduction of human corneas with high doses (5 x 10(7)-3 x 10(8) pfu) of AdV caused eGFP expression in 12-100% of corneal endothelial cells. Corneas were efficiently transduced following up to 28 days in cold storage. Very high AdV doses (2 x 10(9) pfu) reduced endothelial cell densities to 98 (SD 129) nuclei/mm(2) (compared to 2114 (716) nuclei/mm(2) for all other groups). Transgenic protein production peaked at 2.4 (0.9) microg/cornea/day at 2 weeks post-transduction, and decreased to 1.2 (0.4) microg/cornea/day by 33 days, at which time endothelial cell density had decreased to 431 (685) nuclei/mm(2). CONCLUSION: Human corneas can be efficiently transduced by AdV following extended periods of cold storage, and transgene expression is maintained for at least 1 month in vitro.
Influence of advanced recipient and donor age on the outcome of corneal transplantation. Australian Corneal Graft Registry.
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 1997-10) Williams, Keryn Anne; Muehlberg, S M; Lewis, R F; Coster, Douglas John
AIMS: The aims of this study were to examine the influence of advanced recipient and donor age on the long term outcome of corneal transplantation. METHODS: Records of 1036 penetrating corneal grafts in recipients aged > or = 80 years at surgery (defined as the elderly subset) and 8092 donor corneas used for transplantation were obtained from the Australian Corneal Graft Register database, Kaplan-Meier graft survival plots were compared using log rank statistics. RESULTS: Elderly recipients constituted 15% of the recipient pool. The major indication for corneal transplantation in the elderly was bullous keratopathy. Graft survival fell with increasing recipient age (p < 0.00001); the major cause of graft failure was rejection (33%). The desired outcome in 51% of cases was to improve vision and in 42% of cases to relieve pain; 23% of elderly recipients achieved a Snellen acuity of 6/18 or better in the grafted eye and 66% recorded improved acuity after transplantation. Elderly recipients suffered more complications and comorbidities in the grafted eye than did younger recipients. Donor age (stratified in 10 year intervals) did not influence corneal graft survival significantly (p = 0.10). CONCLUSIONS: Elderly graft recipients fared less well after corneal transplantation than did younger recipients, but outcomes in terms of long term graft survival and visual rehabilitation were still good. Donor age did not affect graft survival.
Influence of format on in vitro penetration of antibody fragments through porcine cornea
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2005-09) Brereton, Helen Mary; Taylor, Susan D; Farrall, Alexandra L; Hocking, Dianna M; Thiel, Michael A; Tea, Melinda Nay; Coster, Douglas John; Williams, Keryn Anne
AIM: Antibody fragments, appropriately formulated, can penetrate through the ocular surface and thus have potential as therapeutic agents. The aim was to investigate the influence of protein fragment format on the kinetics and extent of ocular penetration in vitro. METHODS: Immunoglobulin single chain variable domain fragments of a murine monoclonal antibody with specificity for rat CD4 were engineered with a 20 or 11 amino acid linker by assembly polymerase chain reaction, expressed in Escherichia coli and purified by chromatography. Fab fragments of the parental antibody were prepared by papain digestion. Antibody fragments were formulated with a penetration and a viscosity enhancer and were applied to the surface of perfused pig corneas for up to 10 hours in vitro. Penetration was quantified by flow cytometry on rat thymocytes. RESULTS: 20-mer antibody fragments formed natural monomers and dimers following purification that could be separately isolated, while 11-mer fragments were dimeric. All formats of fragment (20-mer monomers and dimers, 11-mer dimers, Fab) showed penetration through the pig cornea after 6 hours of intermittent topical administration. CONCLUSION: Antibody fragments of different shapes and sizes can penetrate the cornea after topical administration, thereby increasing the potential of this class of proteins for topical ophthalmic use.
An instrument for assessment of subjective visual disability in cataract patients
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 1998) Pesudovs, Konrad; Coster, Douglas John
The construction and validation of an instrument for the assessment of subjective visual disability in the cataract patient is described. This instrument is specifically designed for measuring the outcome of cataract surgery with respect to visual disability. METHODS: Visually related activities thought to be affected by cataract were considered for the questionnaire. These were reduced by pilot study and principal components analysis to 18 items. A patient's assessment of his/her ability to perform each task was scored on a four point scale. Scores were averaged to create an overall index of visual disability, as well as subscale indices for mobility related disability, distance/lighting/reading related disability, and near and related tasks visual disability. The questionnaire, administered verbally is entitled "The Visual Disability Assessment (VDA)". Reliability testing included test-retest reliability, interobserver reliability (p, the intraclass correlation coefficient), and internal consistency reliability (Cronbach's alpha). Construct validation, the process for proving that a test measures what it is supposed to measure, included consideration of content validity, comparison with the established Activities of Daily Vision Scale (ADVS) and empirical support with factor analysis. RESULTS: For the four indices, interobserver reliability varied from 0.92 to 0.94, test-retest reliability varied from 0.96 to 0.98, and internal consistency reliability varied from 0.80 to 0.93. The VDA compared favourably with the ADVS by correlation, but Bland-Altman analysis demonstrated that the two instruments were not clinically interchangeable. Factor analysis suggests that all test items measure a common theme, and the subgroupings reflect common themes. CONCLUSIONS: The VDA is easy to administer because it has a short test time and scoring is straightforward. It has excellent interobserver, test-retest, and internal consistency reliability, and compares favourably with the ADVS, another test of visual disability. Factor analysis demonstrated that the 18 items measure a related theme, which can be assumed to be visual disability. The VDA is a valid instrument which provides a comprehensive assessment of visual disability in cataract patients and is designed to detect changes within a patient over time.
Late onset post-keratoplasty astigmatism in patients with keratoconus
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2004-03) Lim, L; Pesudovs, Konrad; Goggin, M; Coster, Douglas John
AIM: 10 eyes of 10 patients are reported where progression of keratoconus in the host cornea occurred more than 10 years after penetrating keratoplasty with resultant increase in astigmatism. The technique and results of graft refractive surgery in seven eyes are presented. METHODS: The clinical features and management of these patients were retrospectively analysed. Graft refractive surgery involved an incision at the graft-host junction adjacent to the host thinning with compressive resuturing. Astigmatic changes were calculated using vector analysis. RESULTS: There were seven men and three women with a mean age of 41.2 years. The average age when undergoing penetrating keratoplasty in the affected eye was 28.4 years and the average time after penetrating keratoplasty until keratoconus appeared in the host cornea defined by host thinning was 13.5 years. The mean cylinder power before host thinning was noted was 5.07 D (SD 2.19) and the mean after host thinning was 11.0 D (2.53). The mean vector calculated disease induced astigmatism magnitude was 7.59 D (3.09). Graft refractive surgery was performed in seven eyes. The mean cylinder power before and after graft refractive surgery was 11.28 D (2.15) and 7.09 D (5.53) respectively. The surgically induced astigmatism vector magnitude was 7.36 D (4.88). CONCLUSION: Progression of keratoconus in the host cornea late after penetrating keratoplasty is characterised by a large astigmatic change where the flat axis of astigmatism passes through an area of host thinning visible on slit lamp examination. Compressive resuturing performed in the area of host thinning resulted in satisfactory reduction of astigmatism.
The long term outcome of limbal allografts: the search for surviving cells
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2001-05) Henderson, Timothy R; Coster, Douglas John; Williams, Keryn Anne
BACKGROUND/AIMS: Limbal allotransplantation is increasingly being used for ocular surface repair in patients with limbal stem cell dysfunction. However, it is uncertain whether donor cells survive long term on the ocular surface and whether patients maintain the early benefits of the procedure. The aims of this study were to investigate the long term outcome of clinical limbal allografts and to correlate outcome with donor cell survival. METHODS: Five patients who had undergone allotransplantation-four keratolimbal allografts and one tarsoconjunctival allograft-from 3-5 years previously, and for whom residual frozen donor ocular tissue was available, were reviewed. Survival of donor cells lifted from the recipient ocular surface by impression cytology was investigated by DNA fingerprinting using primers detecting variable nucleotide tandem repeat sequences. Recipient buccal cells and scleral samples from the remnant donor eye were used to genotype recipients and donors, respectively. Polymerase chain reaction products were sized by Genescan analysis. RESULTS: An objective long term benefit from the procedure (improved Snellen acuity, reduced frequency of epithelial defects, reduced vascularisation, and scarring) was recorded for four patients. Some subjective benefit was also reported. However, in no instances were donor cells recovered from the ocular surface at 3-5 years post-graft. Initial experiments to examine sensitivity indicated that any surviving donor cells must have constituted less than 2.5% of cells sampled. CONCLUSION: Limbal stem cell allotransplantation can provide long term benefits, as measured by objective criteria. However, such benefits do not necessarily correlate with survival of measurable numbers of donor cells on the ocular surface.
Management of high-risk corneal grafts
(2003) Williams, Keryn Anne; Coster, Douglas John
Ophthalmic nurse practitioner led diabetic retinopathy screening. Results of a 3-month trial
(2006) Kirkwood, Bradley J; Coster, Douglas John; Essex, Rohan
Pythium insidiosum keratitis confirmed by DNA sequence analysis
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2001-04) Badenoch, Paul Raymond; Coster, Douglas John; Wetherall, Bruce Lawrence; Brettig, Helen T; Rozenbilds, Maris A M; Drenth, Andre; Wagels, Gabriele
Case report/letter detailing identification of the pathogen pythium insidiosum by nucleic acid sequencing.
A simple corneal perfusion chamber for drug penetration and toxicity studies
(BMJ Publishing Group - http://bjo.bmjjournals.com/, 2001-04) Thiel, Michael A; Morlet, N; Schulz, D; Edelhauser, HF; Dart, JK; Coster, Douglas John; Williams, Keryn Anne
AIMS: Corneal perfusion chambers are important tools in the development and assessment of ophthalmic drugs. The aim of this study was to design and test a modified perfusion chamber suitable for topical application of drugs to isolated corneoscleral preparations, and which allowed continuous monitoring of endothelial cell function. METHODS: A polycarbonate and stainless steel perfusion chamber was designed to clamp corneas in a horizontal plane suitable for topical drug delivery. Endothelial cell function was assessed by ultrasonic pachymetry and specular microscopy during perfusion. Epithelial barrier function was assessed by penetration of fluorescein. Leakage was examined by measuring penetration of a large protein, IgG. Tissue architecture after perfusion was examined by conventional histology. RESULTS: Corneas maintained a functionally and morphologically intact endothelial monolayer during perfusion periods of up to 14 hours. The epithelial barrier function was well preserved. The tissue clamp sealed the preparation effectively against leakage of macromolecules. CONCLUSION: The new chamber device forms a reliable tool for in vitro drug penetration and toxicity studies in isolated perfused corneoscleral tissue.
Stability of housekeeping gene expression in the rat retina during exposure to cyclic hyperoxia
(2007) Williams, Keryn Anne; van Wijngaarden, Peter; Brereton, Helen Mary; Coster, Douglas John

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