A 40-year-old man, with a history of penetrating corneal injury as a child, underwent femtosecond-enabled dovetail keratoplasty, anterior vitrectomy and secondary intraocular lens suturing to repair his corneal scar and aphakia. A partial thickness dovetail pattern was performed in the recipient cornea using the femotsecond laser. The posterior side-cut was initiated ~100 µm anterior to the Descemet membrane and extended obliquely towards the outer edge of a ring lamellar cut, positioned at ~300 µm stromal depth. The anterior side-cut was extended from the internal edge of the ring lamellar cut to the corneal surface. Using an artificial chamber, the femtosecond laser was used to create a full-thickness 0.2 mm oversized femtosecond-enabled dovetail trephination with similar anterior lamellar depth (~300 µm). Wound closure, using interrupted 10–0 nylon sutures, was guided by preplaced radial alignment laser microincisions and tongue-in-groove midstromal suture positioning.

Excellent alignment and stability of the donor and recipient tissue were observed immediately postoperatively and 5 months after surgery. The feasibility of the "dovetail" pattern of PK and the tongue-in-groove suture positioning is demonstrated.

Case reports.

Prompt regression of conjunctival microvessels in the pterygial bed was documented 1 week after a single subconjunctival injection of ranibizumab (one case) or bevacizumab (two cases). No side-effects were noted over 13 months of follow-up in the first case, 6 months in the second case and 1 month in the third case.

Selective blockade of vascular endothelial growth factor was effective in causing regression of conjunctival microvessels in three eyes with inflamed pterygium or residual pterygia.

Trachoma Rapid Assessment methodology was used in Kiribati, Nauru, Vanuatu, Solomon Islands and Fiji. Advised by key informants, high-risk communities were chosen from each country. All available children aged 1–9 years and adults >=40 years were examined.

A total of 903 adults >=40 years and 3102 children aged 1–9 years were screened at 67 sites. Rates of active trachoma in children were >15% in all sites in Kiribati and >20% in all sites in Nauru. However, there was a high variability of rates of active trachoma in survey sites in Vanuatu, Solomon Islands and Fiji with rates ranging from 0% to 43% (average 23.3%), 6.0% to 51.9% (average 30.5%) and 0% to 48.8% (average 22.1%) respectively. Average rates of scarring trachoma in adults were 61.9% in Kiribati, 12.5% in Nauru, 38.2% in Vanuatu, 67.0% in the Solomon Islands and 18.8% in Fiji. Rates of trichiasis and trichiasis surgeries suggest the possibility of blinding trachoma in the region.

The findings indicate that trachoma is present in all the Pacific Island countries screened. Further prevalence studies are required, and trachoma control measures should be considered.

A cross-sectional study. Utility values were measured using the time trade-off (TTO) and standard gamble (gamble for blindness) methods. Standard face-to-face interviews were conducted. Results of ophthalmic examinations were recorded.

One hundred and six patients were recruited, 16 (15.1%) male and 90 (84.9%) female, mean age 63.21 years (SD 7.46). Fifty-five (50.9%) patients had a history of acute angle closure (AAC). The mean duration from glaucoma diagnosed to the day of interview was 7.69 (3.30) months. All participants underwent laser therapy, and 12 patients (11.3%) also received trabeculectomies. The mean utility value was 0.75 (0.14) (95% CI 0.73 to 0.78) with the TTO technique and 0.80 (0.08) (95% CI 0.78 to 0.81) with the standard gamble (SG) technique. The correlation between the two values was significant (p<0.001). Age, educational status, better best-corrected visual acuity (BCVA), worse BCVA and better visual field had significant effects on the utility values. Patients’ self-evaluation of the impact of PAC/G upon their lives correlated with the TTO utility values (p = 0.008) and not with the SG utility values (p = 0.652).

The mean TTO utility score of the studied population is 0.75. The utility value is directly dependent on age, the degree of visual function loss and educational status. It is likely that patients find the TTO method easier to comprehend than the SG method.

Cases were identified through surveys, outreach and clinics. They underwent preoperative visual acuity measurement and ophthalmic examination. Cases were re-examined 8–15 months after cataract surgery. Information on age, gender, poverty and literacy was collected at baseline.

452 eyes of 346 people underwent surgery. 124 (27%) eyes had an adverse outcome. In Kenya and the Philippines, the main cause of adverse outcome was refractive error (37% and 49% respectively of all adverse outcomes) then comorbid ocular disease (26% and 27%). In Bangladesh, this was comorbid disease (58%) then surgical complications (21%). There was no significant association between adverse outcome and gender, age, literacy, poverty or preoperative visual acuity.

Adverse outcomes following cataract surgery were frequent in the three countries. Main causes were refractive error and preoperative comorbidities. Many patients are not attaining the outcomes available with modern surgery. Focus should be on correcting refractive error, through operative techniques or postoperative refraction, and on a system for assessing comorbidities and communicating risk to patients. These are only achievable with a commitment to ongoing surgical audit.

A retrospective study of 531 manuscripts, involving 1182 masked reviews, submitted to Journal of American Association for Pediatric Ophthalmology and Strabismus from 2000 to 2005.

Data were extracted from recommendation forms completed by each referee during review. Investigated variables consisted of reviewer’s knowledge of author identity, recommendation (accept, revise, or reject publishing), eventual manuscript status (published or not), review quality, gender, country, medical practice setting (academic or private) and editorial board status.

This study involved the largest number of manuscripts ever used to evaluate the importance of author masking. Reviewer’s knowledge of the author’s identity had no effect on review quality. However, proportionally fewer manuscripts were published when there was no idea of the author’s identity, compared with when it was allegedly known or suspected (p<0.0001). Manuscripts had lower recommendation scores when there was no idea of the author’s identity compared with when allegedly known (p = 0.0001) or suspected (p = 0.004).

Reviewers were more favourable when they allegedly knew or suspected the author’s identity. Double-masking may improve the quality of biomedical publishing or at least reduce reviewer bias for effectively masked manuscripts.

The RB database was used to identify children with heritable, bilateral RB treated with primary chemotherapy (six cycles of vincristine, etoposide and carboplatin). Only Group D eyes with more than 12 months’ follow-up were analysed. The timing, number and type of salvage treatments were recorded. Kaplan–Meier estimates for the ocular survival and event-free survival (percentage of eyes that avoided external beam radiotherapy and/or enucleation) were performed as a function of time.

Of 18 group D eyes, two (11%) were treated successfully with chemotherapy alone, nine (50%) underwent successful salvage treatment, and seven (39%) were enucleated. The median time from completing chemotherapy to enucleation was 9 months (range 4 to 25 months). Ocular survival was 67% at 2 years. External beam radiotherapy proved successful salvage treatment in five of nine eyes, so the event-free survival was 34% at 2 years.

Multiagent chemotherapy alone is rarely sufficient for the preservation of group D eyes. External beam radiotherapy and plaque radiotherapy remain important salvage treatments for advanced, heritable retinoblastoma.

To study the effectiveness of EBRT as a salvage treatment after failed primary chemotherapy and focal treatment in bilateral retinoblastoma.

This is a retrospective observational case series. The outcome measures after EBRT are: rate of eye preservation, rate of tumour control, visual potential, visual acuity and radiation-induced side-effects.

Thirty-six eyes (22 patients) were included. The median follow-up after EBRT was 40 months (19–165 months). Thirty-two eyes received lens-sparing radiotherapy, and four received whole-eye radiation. The rate of eye preservation was 83.3% (30/36 eyes). Twenty-four eyes (66.7%) were controlled by EBRT and required no further treatment. Of the 30 preserved eyes, 20 eyes (66.7%) had extramacular tumours without retinal detachment and therefore potential for central vision. The final visual acuity was recorded for 19 eyes. Ten eyes (52.6%) read 6/9–6/5, three eyes (15.8%) read 6/18–6/36, and six eyes (31.6%) read 6/60 or worse. Significant radiation- induced side effects were limited to cataracts and dry eyes with whole-eye radiation. There were no second cancers or deaths.

Salvage EBRT is highly effective in preserving eyes with useful vision in bilateral retinoblastoma after failed chemotherapy and focal treatments. These results will help the parents and ophthalmologists of such patients to reach an informed decision when weighing up the benefits of EBRT against its potential oncogenic effect.

Peripapillary RNFL thickness was measured using the standard "fast" RNFL scan-protocol and proportional 2.27xdisc scan protocol. Disc size was measured using the "fast" optic disc protocol. The correlation between RNFL thickness for each scan-protocol and disc size was analysed.

In normal eyes, RNFL thickness was independent of the optic-disc area using the fixed-diameter protocol (p = 0.92) but was inversely proportional to disc size using the proportional protocol (p<0.001). In glaucoma suspects, the optic-disc area correlated with RNFL thickness using the fixed-diameter protocol (p<0.001). In the multivariate analysis in glaucomatous eyes, the RNFL thickness using the fixed-diameter protocol was significantly affected by the mean deviation on visual fields but not by disc area (p<0.001 and p = 0.64 respectively)

In normal subjects, disc size does not appear to affect RNFL measurements by OCT using the fixed-diameter protocol, thus indicating that RNFL thickness may be related to distance from the centre of the optic disc rather than the margin. The thicker RNFL observed in large glaucomatous discs in this study may be related to the earlier stage of glaucoma, though it may not apply to the general population.

This study involved 70 eyes (45 patients) with clinically significant macular oedema that underwent focal laser photocoagulation using the Early Treatment Diabetic Retinopathy Study protocol. Preoperative macular OCT images were retrospectively examined. OCT features were classified into four patterns: diffuse retinal thickening (DRT); cystoid macular oedema (CMO), serous retinal detachment and vitreomacular interface abnormalities (VMIA). Changes in retinal thickness, retinal volume and visual acuity (VA) after focal laser photocoagulation were evaluated and compared with respect to their OCT features.

After focal laser photocoagulation, changes in retinal thickness and retinal volume were significantly different for different OCT types (p = 0.002 and p<0.001). The change in VA from baseline was not significantly different between groups (p = 0.613). The DRT pattern was associated with a greater reduction in retinal thickening and better VA improvement than the CMO or VMIA patterns. Proportions of patients with persistent DMO (central macular thickness >250 µm after laser treatment) were greater for the CMO and VMIA patterns than DRT pattern.

DRT patients achieved a greater reduction in retinal thickening and greater VA increases than CMO and VMIA patients. We suggest that classifying DMO structural patterns using OCT might allow visual outcome to be predicted after laser photocoagulation.

Retrospective analysis of an interventional case series. IFN alpha-2a was administered at an initial dose of 3 or 6 million IU per day subcutaneously and tapered afterwards to the lowest possible dose to maintain the absence of CMO. Treatment efficacy was assessed by optical coherence tomography.

Twenty-four patients with chronic CMO (median duration 36.0 months) due to non-infectious anterior (n = 2), intermediate (n = 18) or posterior (n = 4) uveitis have been analysed. Ineffective pretreatment included systemic corticosteroids (all patients), acetazolamide (22 patients) and at least one immunosuppressive drug (18 patients). IFN therapy was shown to be effective ( = complete resolution of CMO within 3 months, able to taper IFN) in 15 patients (62.5%), partly effective ( = incomplete resolution of CMO, unable to taper IFN) in six patients (25.0%) and not effective ( = no response or recurrence of CMO) in three patients (12.5%). IFN treatment was generally well tolerated. Common side effects including flu-like symptoms, fatigue or increased liver enzymes were dose-dependent and led to discontinuation of IFN in only one patient.

The data demonstrate IFN alpha to be an effective and well-tolerated therapy for chronic refractory uveitic CMO.

Subjects with fibrovascular pigment epithelial detachment, subfoveal choroidal neovascularisation extending under the geometric centre of the foveal avascular zone and/or macular thickness of at least 300 µm in both eyes were included. Sixteen eyes were included and randomised equally to receive either three infusions of 5 mg/kg Avastin or 100 ml of 0.9% sodium chloride every 2 weeks. The main outcome measure was the lesion size. The follow-up time was 24 weeks.

Throughout the 24-week follow-up, the lesion size and macular thickness decreased in the Avastin group by 0.5 (SD 0.08) mm and 103.6 (14.9) µl respectively. In both groups, visual acuity remained stable in seven eyes and decreased in one eye. At the end of follow-up, 50% of the eyes in the Avastin group became fibrotic, 37.5% remained unchanged, and 12.5% developed a subretinal bleeding. There was a treatable rise in blood pressure after Avastin treatment.

Systemic Avastin could be offered to patients with exudative age-related macular degeneration in both eyes and/or patients who refuse intravitreal injections if blood pressure is normal and there is no history of thrombosis.

Five affected children of a consanguineous Moroccan family were investigated by ophthalmic examinations, including fundus photography, autofluorescence (FAF) imaging, optical coherence tomography (OCT), psychophysical and electrophysiological methods.

Affected children were between 5 and 19 years of age, allowing an estimation of disease progression. Electroretinography demonstrated loss of scotopic and photopic function in the first decade of life. Younger siblings showed drusen-like deposits with focal relatively increased FAF in the macular area. With increasing age, a yellowish lesion with relatively increased FAF and subsequent macular atrophy developed. Visual acuity deteriorated with age and ranged between 20/50 in the best eye of the youngest affected and 20/400 in the worst eye of the oldest affected sibling. Spectral-domain OCT revealed debris-like material in the subneurosensory space.

The splice site mutation c.2189+1G>T in MERTK causes rod–cone dystrophy with a distinct macular phenotype. The debris in the subneurosensory space resembles that in the Royal College of Surgeons (RCS) rat being the mertk animal model. Patients might therefore benefit from advances in gene therapy that were previously achieved in the RCS rat.

This is an interventional case series of 13 members from a non-consanguineous Chinese family. All patients underwent complete ophthalmological examination and slit-lamp photography. Subsequent corneal transplantations were performed (n = 3). Blood samples were taken for DNA extraction and subsequent genetic analysis.

Genotyping indicated linkage to the locus at chromosome 1p36. Screening of the UBIAD1 gene identified a highly conserved mutation, Ser171Pro. Phenotypic variation in this large pedigree is similar to that seen in Caucasian patients. Surgical management of patients with anterior lamellar keratoplasty and deep anterior lamellar keratoplasty showed good visual outcomes.

The S171P mutation is described for the first time in a Chinese family. This is the largest non-Caucasian pedigree described with SCD. Visual rehabilitation may be performed successfully with lamellar surgical procedures as opposed to full-thickness corneal grafts.

The genotype of 41 affected members of 16 families and nine sporadic cases was investigated by direct sequencing of the TGFBI gene. Clinical, histological and immunohistochemical characteristics of corneal opacification were reported and compared with the coding region changes in the TGFBI gene.

A novel mutation Leu509Pro was detected in one family with a geographic pattern-like clinical phenotype. Histopathologically we found amyloid together with non-amyloid deposits and immunohistochemical staining of Keratoepithelin (KE) KE2 and KE15 antibodies. In two families and one sporadic case the novel mutation Gly623Arg with a late-onset, map-like corneal dystrophy was identified. Here amyloid and immunohistochemical staining of only KE2 antibodies occurred. Further, five already known mutations are reported: Arg124Cys Arg555Trp Arg124His His626Arg, Ala546Asp in 13 families and five sporadic cases of German origin. The underlying gene defect within the TBFBI gene was not identified in any of the four probands with Thiel–Behnke corneal dystrophy.

The two novel mutations within the TGFBI gene add another two phenotypes with atypical immunohistochemical and histopathological features to those so far reported.

Nine consecutive patients (18 eyes) with serologically confirmed MT at the acute stage were enrolled in this prospective, non-comparative study. All patients underwent complete ophthalmic examination, including dilated biomicroscopic fundus examination, fundus photography, fluorescein angiography (FA) and indocyanine green (ICG) angiography.

Of nine patients, eight (88.9%) had bilateral ocular involvement related to MT, with (n = 3) or without (n = 5) associated ocular symptoms. Findings included mild vitreous inflammation (10 eyes; 55.6%), white retinal lesions (nine eyes; 50%), retinal haemorrhages (four eyes; 22.2%), retinal vascular leakage (seven eyes; 38.9%), hypofluorescent choroidal dots on FA and/or ICG angiography (11 eyes; 61.1%), optic-disc swelling (two eyes; 11.1%), optic neuritis (one eye; 5.6%) and optic-disc staining (11 eyes; 61.1%). All ocular findings had a self-limited course.

Ocular involvement is frequently observed in acute MT. A systematic fundus examination, complemented by angiography in selected cases, may be helpful in establishing an early clinical diagnosis of the disease while serological testing is pending.

Hospital files were retrospectively examined for 99 Chinese PACG patients; 75 patients underwent combined surgery and 24 underwent trabeculectomy alone. Success rates were assessed with the Kaplan–Meier survival analysis. The main outcome was the complete success rate defined as either a >20% reduction in intraocular pressure (IOP) or an IOP that remained below 15 mm Hg, with no medications required.

Patients in the combined group and trabeculectomy group had a mean follow-up period of 25.8 (SD 10.8) months and 31.4 (8.9) months, respectively. Survival analysis showed that the complete success rate at 3 years was 56% in the combined group and 54% in the trabeculectomy group (p = 0.903). There were no significant differences between groups in either IOP or the number of glaucoma medications throughout the 3-year follow-up. The incidences of postoperative complications were similar between groups (p = 0.232). No additional IOP-lowering surgical procedures were required in the combined group, while 13 (54%) eyes in the trabeculectomy group required either cataract extraction or further IOP-lowering surgical procedures (p <0.001).

In patients with PACG, the long-term IOP-lowering effect and surgical complications of combined trabeculectomy and cataract extraction are comparable with those of trabeculectomy alone. However, the combined surgery incurred fewer subsequent surgical interventions.

Measurements were taken with the LenStar and IOLMaster on 112 patients aged 41–96 years listed for cataract surgery. A subgroup of 21 patients also had A-scan applanation ultrasonography (OcuScan) performed. Intersession repeatability of the LenStar measurements was assessed on 32 patients

LenStar measurements of white-to-white were similar to the IOLMaster (average difference 0.06 (SD 0.03) D; p = 0.305); corneal curvature measurements were similar to the IOLMaster (average difference –0.04 (0.20) D; p = 0.240); anterior chamber depth measurements were significantly longer than the IOLMaster (by 0.10 (0.40) mm) and ultrasound (by 0.32 (0.62) mm; p<0.001); crystalline lens thickness measurements were similar to ultrasound (difference 0.16 (0.83) mm, p = 0.382); axial length measurements were significantly longer than the IOLMaster (by 0.01 (0.02) mm) but shorter than ultrasound (by 0.14 (0.15) mm; p<0.001). The LensStar was unable to take measurements due to dense media opacities in a similar number of patients to the IOLMaster (9–10%). The LenStar biometric measurements were found to be highly repeatable (variability <=2% of average value).

Although there were some statistical differences between ocular biometry measurements between the LenStar and current clinical instruments, they were not clinically significant. LenStar measurements were highly repeatable and the instrument easy to use.

Controlled, randomised multicentre trial.

12 university clinics.

124 patients were randomly assigned to either BR or RR. Standardised protocol prescribed that the total relocation of the muscles, in millimetres, was calculated by dividing the preoperative latent angle of strabismus at distance, in degrees, by 1.6.

Alignment assessed as the variation of the postoperative angle of strabismus during alternating cover.

The mean preoperative latent angle of strabismus at distance fixation was +17.2° (SD 4.4) for BR and +17.5° (4.0) for RR. The mean postoperative angle of strabismus at distance was +2.3° (5.1) for BR and +2.9° (3.5) for RR (p = 0.46 for reduction in the angle and p = 0.22 for the within-group variation). The mean reduction in the angle of strabismus was 1.41° (0.45) per millimetre of muscle relocation for RR and 1.47 (0.50) for BR (p = 0.50 for reduction in the angle). Alignment was associated with postoperative binocular vision (p = 0.001) in both groups.

No statistically significant difference was found between BR and RR as surgery for infantile esotropia.

Sorafenib or vehicle was administered orally to female C57BL/6 mice at the onset (day 0) of experiments. CNV was induced by laser photocoagulation the following day. After 14 days, mice were perfused with fluorescein-labelled dextran, and the area of CNV was measured on choroidal flat mounts by image analysis. In some groups of mice, treatments were started 7 days after the laser photocoagulation to determine the effect of the agent on established CNV. Expression of phosphorylated extracellular signal-regulated kinase (p-ERK) in choroidal tissues was measured by Western blot analysis to demonstrate the kinase-inhibitory effect of sorafenib in intracellular signalling pathways involved in CNV formation.

Sorafenib significantly reduced the extent of CNV in a dose-dependent manner. The area of CNV was reduced by 43% in the 30 mg/kg/day group and by 61% in the 60 mg/kg/day group compared with vehicle-treated controls (both p<0.0001). Oral administration of sorafenib also caused significant regression of established CNV. The area of CNV was reduced by 59% in the 30 mg/kg/day group and by 66% in the 60 mg/kg/day group compared with both baseline and control measurements (p<0.0001). The expression of p-ERK in choroidal tissues was increased within 1 day of laser photocoagulation and remained elevated for 2 weeks. The expression of p-ERK was suppressed by sorafenib.

Sorafenib demonstrated antiangiogenic properties in a mouse model of CNV and may be useful in the treatment of CNV.

A blue-light confocal scanning laser ophthalmoscope (bCSLO, 460 nm excitation and 490 nm detection) was used to image Thy1-cyan fluorescent protein (CFP) mice before and weekly for 4 weeks after transiently elevating the intraocular pressure to 115 mm Hg for 45 min (n = 4) or 90 min (n = 5) to induce ischaemic injury. Corresponding retinal areas before and after the intraocular pressure (IOP) elevation, during the period of ischaemic reperfusion, were compared, and the fluorescent spots (Thy-1 expressing RGCs) were counted. The longitudinal profile of CFP-expressing RGCs was modelled with a linear regression equation. The spatial distribution of RGC damage was analysed in the superior, nasal, inferior and temporal quadrants of the retina.

No significant change was found at 4 weeks after 45 min of IOP elevation (n = 4, p = 0.465). The average RGC densities before and 4 weeks after IOP elevation were 1660 (SD 242) cells/mm² and 1624 (209) cells/mm², respectively. However, significant loss of CFP-expressing RGCs was detected at 1 week following 90 min of IOP elevation (n = 5, p<0.001). After this initial RGC loss, no significant change was detected subsequently. The proportion of RGC fluorescence remaining was variable and ranged from 14.5% to 79.5% at 4 weeks after the IOP elevation. The average RGC densities before and 4 weeks after IOP elevation were 1443 (162) cells/mm² and 680 (385) cells/mm², respectively. Diffuse loss of fluorescent RGCs was observed in the spatial distribution analysis.

The longitudinal profile of Thy-1 expressing RGC fluorescence loss after ischaemic injury is non-progressive and unrelated to the duration of reperfusion.

A histopathological analysis, using confocal microscopy, was performed on six retinal specimens. Results were compared with those from two further retinal specimens, collected during RPE transplantation, to control for the effects of vitrectomy and ARMD. In addition, a third control specimen from a cadaver with no history of ophthalmic disease was also analysed.

In the translocation specimens, rods and cones were relatively well preserved but showed reduced density and outer segment length. In four specimens, there were focal areas of rod opsin redistribution to the inner segment, but this was not observed in the controls. Staining with calbindin was decreased in cones compared with controls but normal in horizontal and amacrine cells. Rod bipolar cells were mildly disorganised, and in one there was evidence of neurite sprouting. Glial fibrillar acidic protein was raised in both translocation and transplantation retinae but not in the cadaver control.

In this study, there was little evidence of cellular injury following iatrogenic detachment; however, the rate of PVR following translocation surgery infers that cellular events set in motion may continue despite early reattachment.

Surgical specimens of conjunctivae with or without pingueculae were obtained from eight patients. Immunohistochemical localisation of d-β-aspartic acid-containing proteins was performed using a polyclonal antibody against d-β-aspartic acid-containing peptides.

Strong immunoreactivity to d-β-aspartic acid-containing peptides was detected in the subepithelial amorphous materials of all surgical specimens with pingueculae. In contrast, no immunoreactivity to d-β-aspartic acid-containing peptides was detected in the specimens without pingueculae.

Pingueculae are thought to be aggregates of proteins that contain d-β-aspartic acid residues. It is known that the conversion of l- to d-aspartyl residues is accelerated by ultraviolet irradiation. In addition, d-β-aspartic acid-containing proteins, in general, tend to aggregate with each other and accumulate in the tissues. These facts indicate that ultraviolet irradiation-induced racemisation of aspartic acid is closely related to the development of pingueculae.

Immunohistochemical localisation of d-β-Asp-containing proteins using polyclonal antibodies raised against d-β-Asp-containing peptides was examined in three corneas with CDK, three corneas with interstitial keratitis, six corneas with bullous keratopathy, and three corneas without any corneal diseases.

Strong immunoreactivity to d-β-Asp-containing peptide was detected in all the surgical specimens with CDK. In contrast, no immunoreactivities to d-β-Asp-containing peptides were detected in the surgical specimens with bullous keratopathy, interstitial keratitis, or no corneal diseases.

CDK was regarded as aggregations of d-β-Asp-containing proteins. The formation of d-amino acids in protein causes the different side chain orientations and β-linkage of Asp residues elongates the main chain of proteins. Therefore, d-β-Asp formation will result in a partial unfolding of proteins leading to the aggregation of proteins seen in CDK.

HLEC cultured on amniotic membranes were stored in organ culture medium in a closed container at 23°C. Sterility of storage media was tested using a Bactec 9240 blood culture instrument (Becton Dickinson, Maryland) for incubation and periodic reading. Viability was analysed by calcein-acetoxymethyl ester/ethidium homodimer-1 assay, morphology by light microscopy and cellular phenotype by immunohistochemistry.

No microbial contamination was observed after 1 week’s storage. Viability of cultured HLEC was 87.9 (SD 6.4)% and 52.7 (13.1)% after 2 and 3 weeks of storage, respectively, compared with 98.8 (2.6)% before storage (p<0.001). The multilayered structure was preserved in 70% of cultures following 2 weeks of storage but lost after 3 weeks. A less differentiated phenotype was maintained.

This study is the first to verify the sterility of HLEC cultures prior to transplantation. Although a slight decrease in viability was observed following 2 weeks of storage, the HLEC sheets remain acceptable, whereas 3 week’s storage was unsatisfactory.