Application of high intensity focused ultrasound for treatment of open-angle glaucoma in Indian patients.

Purpose: The aim of this study is to assess the efficacy of ultrasound cycloplasty (UCP) in Indian patients with open angle glaucoma (OAG). Methods: A prospective interventional study was designed to treat 73 eyes of 73 patients with OAG with the EyeOP1 device equipped with six miniaturized cylindrical piezoelectric transducers. Two treatment protocols of ultrasound delivery depending on exposure time (8 s and 10 s of shot per transducer) were used. Complete ophthalmic examination, ocular biometry and anterior segment optical coherence tomography were performed preoperatively and patients were followed up at day 1, day 7 and months 1, 2, 3, 6, and 12. The primary outcome measure was "successful" intraocular pressure (IOP) control defined as IOP reduction ≥20% from baseline and IOP value >5 mm Hg at the last follow-up visit. Secondary outcomes were the occurrence of complications and mean IOP during the follow-up period. Results: In all patients, the mean IOP reduced from 23.5 ± 3.0 mmHg before treatment to 15.7 ± 5.4 mmHg at 12 months (P < 0.05). Successful IOP control after a single procedure was 78.3% (79% and 78% in the 8 s and 10 s groups, respectively) at 12 months. Overall, the mean IOP reduction achieved in responding patients was 41% (standard deviation = 12%). Notwithstanding minor side effects such as transient pain, anterior chamber reaction, and refractive error changes, no major intraoperative or postoperative complications (severe hypotony or phthisis) were observed during the follow-up. Conclusion: Our short-term results reveal that UCP is a simple, safe, and noninvasive procedure which enables to significantly reduce the IOP in patients with OAG. The study results in Indian eyes corroborate findings in earlier studies on Caucasian eyes.

No loss of chance of diabetic retinopathy screening by endocrinologists with a digital fundus camera.

OBJECTIVE: To compare the efficacy of the diabetic retinopathy (DR) screening with digital camera by endocrinologists with that by specialist and resident ophthalmologists in terms of sensitivity, specificity, and level of "loss of chance." RESEARCH DESIGN AND METHODS: In a cross-sectional study, 500 adult diabetic patients (1,000 eyes) underwent three-field retinal photography with a digital fundus camera following pupillary dilatation. Five endocrinologists and two ophthalmology residents underwent 40 h of training on screening and grading of DR and detection of associated retinal findings. A κ test compared the accuracy of endocrinologist and ophthalmology resident screening with that performed by experienced ophthalmologists. Screening efficiency of endocrinologists was evaluated in terms of "loss of chance," i.e., missed diagnoses that required ophthalmologist referrals. RESULTS: The mean weighted κ of DR screening performed by endocronologists was similar to that of ophthalmology residents (0.65 vs. 0.73). Out of 456 DR eyes, both endocrinologists and ophthalmology residents misdiagnosed only stage 1 DR (36 and 14, respectively), which did not require ophthalmologist referral. There were no significant differences between endocrinologists and ophthalmology residents in terms of diabetic maculopathy and incidental findings except for papillary cupping and choroidal lesions, which were not the main purpose of the study or of the training. CONCLUSIONS: The endocrinologist with specific training for DR detection using a three-field digital fundus camera with pupillary dilatation can perform a reliable DR screening without any loss of chance for the patients when compared with identical evaluation performed by experienced ophthalmologists.

Comparison of two semiautomated methods for evaluating endothelial cells of eye bank corneas.

PURPOSE: To compare two semiautomated methods of evaluating endothelial cells of eye bank corneas. METHODS: Using a commercially available semiautomatic endothelial analyzer, seven observers determined the endothelial cell density (ECD), coefficient of variation (CV) of cell area, and the percentage of hexagonal cells (hexagonality) of the light microscopic images of the endothelium of 30 organ-cultured corneas. The image quality was graded as good, average, and poor. Border (contour detection and manual retouch) and center (indicating cell centers) methods for identifying endothelial cells were compared. The interobserver variability in ECD determination (indicating reproducibility) and morphometry was statistically analyzed by using the two methods. The importance of accurate pointing of cell centers was assessed by counting on 10 standard photolithographic mosaics and noting the time taken. RESULTS: There was no significant difference in the interobserver variability or between ECDs obtained by the border and center methods. Decrease in image quality had a similar influence on both methods. Although measurement of hexagonality was acceptable by both methods, the CV was reliable only with the border method, with a significant underestimation by the center METHOD: However, an accurate indication of cell center slightly improved the CV estimation. CONCLUSIONS: Although both the border and center methods of semiautomatic evaluation of eye bank corneas measure similar ECD with a similar reproducibility, only the border method gives a reliable morphometry.

Reproducibility of endothelial assessment during corneal organ culture: comparison of a computer-assisted analyzer with manual methods.

PURPOSE: To investigate the reproducibility of endothelial assessment of organ-cultured corneas with the computer-assisted Sambacornea analyzer in comparison with manual METHODS: methods. Seven observers of two eye banks determined the endothelial cell density (ECD) of 30 corneas through a grid overlay placed on endothelial photographs using two manual modes, unaided (naked-eye) and pointing (point-out). ECD was measured with the analyzer, first in automated mode, where analysis was completely machine determined, and then in touched-up mode, where the observer selected the analysis zone and corrected poorly drawn cell borders. Interobserver variability of ECD for the different methods was compared. Reproducibility of morphometry parameters was determined for the touched-up mode. RESULTS: Interobserver variability was +/-19.2% (95% confidence interval [CI], 13.0-25.4) and +/-17.6% (95% CI, 11.9-23.3) for the naked-eye and point-out mode, respectively, whereas the touched-up mode gave the least variability of +/-9.6% (95% CI, 6.5-12.7), confirmed by the highest intraclass correlation coefficient of 0.95 (95% CI, 0.91-0.97). Interobserver variability increased with worsening image quality. Manual modes underestimated ECD (naked-eye by a mean 10.7% [SD, 2.9%]; point-out by a mean 6.9% [SD, 2.3%]), whereas the automated mode overestimated ECD by a mean 14.7% (SD, 24.3%). Reproducibility of morphometric parameters by the touched-up mode was acceptable but was influenced by endothelial pleomorphism. CONCLUSIONS: Manual counting shows systematic underestimation of ECD with high interobserver variability. The analyzer in automated mode overestimates ECD and is absolutely unreliable. Detection of cell contours by the specific algorithm, combined with manual correction by a skilled technician, appears to be the most reliable method of ECD and morphometry determination.

Standard microlithographic mosaics to assess endothelial cell counting methods by light microscopy in eye banks using organ culture.

PURPOSE: To develop standard microscopic hexagonal mosaics mimicking the human corneal endothelium for quality control of endothelial cell density (ECD) measurement and verification of cell counting strategy by light microscopy in eye banks using organ culture. METHODS: A standard slide, the Keratotest, was developed with 10 laser-engraved mosaics and different predetermined "cell" densities representing the range of ECDs observed routinely. Horizontal and vertical micrometric scales were etched adjacently to each mosaic, and a standard microscopy resolution test pattern was included. The Keratotest was applied to assess the reliability of a computer-assisted analyzer developed for corneal endothelial evaluation based on light microscopy images. RESULTS: The Keratotest consisted of 10 microlithographic homogeneous mosaics of 1-mm2 printed area and 1.2-microm cell boundary thickness. The micrometric scale associated with each mosaic aided in simultaneous verification of microscope calibration, and the test pattern aided in checking the microscope resolution. The design was unalterable and reproducible, and the glass slide incorporated in a carbon fiber support ensured easy handling and safe transport. Evaluation of the Keratotest mosaics by the computer-assisted analyzer found a high level of agreement (error margin between +0.12 and -0.46%) with the laser-engraved cell density. CONCLUSIONS: This prototype device enabled assessment of reliability of ECD measurement in eye banks. It also allowed verification of the calibration and resolution of light microscopes. Periodic validation of counting procedure in eye banks with mosaics of known "cell" densities should be useful for standardization of donor corneal tissue quality control.

Experimental study of the survival of metastatic cancer cells in corneal organ culture.

PURPOSE: Transmission of donor malignancy to the recipient could be one of the most disastrous complications of corneal grafting. Because of the scarcity of donor tissue and the lack of sufficient scientific evidence, the harvest of donor tissues from deaths due to systemic malignancy is permitted. This study was conducted to investigate the possible transmission of donor metastatic disease via corneal tissue preserved in organ culture (OC) conditions. METHODS: The viability of four frequent human cancer cell lines (lung, breast, skin, and colon) was studied in OC. Various inoculums of cancer cells labeled with the membrane marker PKH67 were seeded on donor corneas and preserved in OC, followed by cell-tracking studies, histopathology, and immunohistochemistry. HLA matching of the dissected Descemet's membrane (DM) of preserved corneas was conducted, to demonstrate cell adherence. Primary cell culture was performed to confirm the viability of adherent tumor cells. RESULTS: Viability tests showed a poor but persistent survival of cancer cells after 2 weeks in OC. Cell tracking, histopathology, and immunohistochemistry demonstrated cancer cell adherence to donor endothelium. HLA typing of the DM of preserved corneas revealed the presence of cancer cell alleles. Primary culture of the DM showed cell proliferation that was identical with the original cancer cell line, according to HLA studies. CONCLUSIONS: The findings demonstrate that under laboratory conditions, metastatic cancer cells adhere to donor corneal tissue, survive, and retain proliferative capacity during storage in OC. Cell lines differ in their viability potential, as well as the pattern of adherence to donor endothelium. Further in vivo experimentation in laboratory animals is need to determine the safety of such harvests.

Screening for diabetic retinopathy by ophthalmologists and endocrinologists with pupillary dilation and a nonmydriatic digital camera.

PURPOSE: To assess the effects of pupillary dilation on image quality and certitude of screening diabetic retinopathy by trained endocrinologists using a digital nonmydriatic camera. DESIGN: Prospective, comparative, observational case series. METHODS: One hundred fifty patients with diabetes attending a hospital-based photographic screening clinic had five-field (45 degree) digital retinal imaging and mosaic construction, first through dark-adapted pupils, then after a single application of tropicamide 1%. The 600 sets of images (150 patients, 300 eyes, before and after dilation) were graded independently and in a blinded manner by two endocrinologists and two ophthalmologists, with the consensual opinion on dilated images of the latter serving as the gold standard. The criteria of evaluation were assessment of image quality and certitude of detecting and grading retinopathy. RESULTS: Of 300 eyes, pharmacological mydriasis improved image quality, with an increase in the number of eyes with five good images from seven to 160 and good-quality mosaics from 99 to 233. Dilation allowed better identification of maculopathy in the second eye because there was a decrease in ungradeable central images from 127 to 15 eyes. Dilation increased the number of eyes having retinopathy detected with certainty from 153 to 252 and graded with certainty from 173 to 277. No adverse effects were recorded. CONCLUSIONS: Single application of tropicamide 1% significantly improves image quality and certitude of screening diabetic retinopathy with nonmydriatic camera by endocrinologists.