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Purpose: To describe symmetry of response in fellow eyes of patients undergoing photorefractive keratectomy (PRK) for myopia, analyse the risk factors leading to asymmetry in response and to determine if delayed treatment of the second eye increases safety and predictability of PRK. Methods: Retrospective review of case records of 133 patients who underwent bilateral myopic PRK and had a minimum follow up of 6 months in both eyes. Results: Postoperative uncorrected visual acuity, spherical equivalent (SE) refraction within ±1D of emmetropia, best-corrected visual acuity (BCVA) and corneal haze were not significantly different in fellow eyes of patients undergoing PRK for myopia. Of 87 eyes in group 1 (myopia Conclusion: This study demonstrates that considerable symmetry of response exists in fellow eyes of patients undergoing myopic PRK. Early PRK in the fellow eye of patients with < 6 D myopia is safe and allows quick visual rehabilitation of the patient. In patients with myopia ≥6D, a 3-month interval before treating the second eye may improve the safety of the procedure.

Purpose: To report the timing of retreatment and clinical outcomes in patients with a partial corneal flap during laser in-situ keratomileusis (LASIK), and to describe the causes of this complication. Methods: Retrospective review of case records of four patients (4 eyes) who had a partial corneal flap during LASIK. Results: The mean age of the four patients was 23 ± 4.7 years, and mean preoperative spherical equivalent (SE) refraction was -9.1 ± 3.1D (range, -5.5 to -13D). A 160 μm corneal flap was attempted during the initial treatment. Retreatment with a 180 μm corneal flap was performed at a mean of 5.1 ± 1.6 weeks (range, 4 to 7.5 weeks) after the initial procedure. There were no intraoperative complications during retreatment. Post-LASIK mean SE refraction was -1.0 ± 1.1D (range, +0.38 to -2.0D), after a mean follow up of 19 ± 15.7 weeks (range, 7 to 42 weeks). Best spectacle-corrected visual acuity decreased in one eye from 6/5 to 6/6 and was maintained in the others. Conclusion: LASIK retreatment can be performed as early as one month after a partial flap, if the refraction is stable and a thicker corneal flap is created.

Purpose: To describe clinical features and treatment outcomes in patients with advanced Nocardia asteroides keratitis. Methods: Retrospective review of case records of 7 patients with culture-proven Nocardia keratitis. Results: Corneal infection occurred after corneal trauma in two patients, cataract surgery in three patients, penetrating keratoplasty in one patient and was associated with a silicone buckle element infection in one patient. Mean duration of infection at presentation was 33.4 days (7-75 days), and five patients had received prior treatment with corticosteroids. Six of seven patients had deep corneal suppuration at the time of: presentation, clinically suggestive of mycotic keratitis. In two patients who had received prolonged corticosteroid therapy (≥ 45 days), the eyes could not be salvaged. Complete resolution of infection was achieved in all 4 eyes treated with topical fortified cefazolin eye drops (50mg / ml).

Our technique of pterygium excision with conjunctival-limbal autografting is described and the safety and efficacy of the procedure in India is analysed. Case records of 51 consecutive patients (53 eyes) who underwent surgery at our institute between November 1992 and September 1994 were retrospectively analysed. Recurrence was defined as fibrovascular tissue crossing the corneoscleral limbus onto clear cornea in the area of previous pterygium excision. 2 (3.8%) of the 53 pterygia (primary 36; recurrent 17) recurred, after a mean follow up of 18.9±12.1 months (range: 1.5-43 months). Both recurrences occurred within a year of follow up, in patients who were ≤40 years of age. No major operative or postoperative complications were encountered. The inclusion of limbal tissue in conjunctival autografts following pterygium excision appears to be essential to ensure low recurrence rates. The technique is safe, simple and inexpensive and is recommended for the management of both primary and recurrent pterygia in Indian eyes.

Background & Objective: The limbus is enriched with the stem cells of corneal epithelium. Auto- and allograft limbal transplantations are effective in restoring the corneal epithelium and inhibiting inflammation and neovascularization. Preserved human amniotic membrane (AM) is now widely used as a substrate for ocular surface reconstruction. The combination of limbal and AM transplantation has been shown to improve the surgical outcome in patients with total limbal stem cell deficiency (LSCD). The purpose of this study was to compare the expression of putative stem cell markers ATP binding cassette protein (ABCG2) and keratinocyte stem cell marker: p63 and differentiation markers. (connexin 43 and keratin 3 / keratin 12) on the limbal epithelial cells cultured over the denuded AM with and without the 3T3 murine fibroblast cells as feeder layer., Methods: Human limbal tissues obtained from the cadaveric donor eyes were cultured over the denuded human amniotic membrane in the presence of mitomycin C treated 3T3 fibroblasts and the cultured cells studied for the expression of ABCG2 and p63 by immunohistochemistry and Western blot. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was done on the cultured cells at varying intervals of time for expression of ABCG2, p63, connexin43 (Cnx43), and keratin 3 (K3) and keratin 12 (K12)., Results: The growth rates were similar in both denuded AM and denuded AM + 3T3. The cells cultured over AM + 3T3 showed the expression of p63 and ABCG2 till 21 days of incubation by immunohistochemistry and Western blot. The expression of p63 and ABCG2 were retained till 21 days of incubation on the cells cultured over denuded AM + 3T3, whereas it was expressed only till day 8 on the cells cultured over the denuded membrane by semi quantitative RT-PCR. Cnx43 and K3/K12 were observed in both the conditions., Interpretation & Conclusion: The limbal epithelial cells cultured in the presence of mitomycin C treated 3T3 feeder layer were able to maintain the expression of putative stem cell markers. Further in vitro studies using feeder layer will enable us to understand the factors, which play a role in maintaining the limbal stem cell niche.

Background & Objectives: The ocular surface is an ideal region to study the epithelial stem cell (SC) biology because of the unique spatial arrangement of stem cells and transient amplifying cells. A major challenge in corneal SC biology is the ability to identify SC in vitro and in situ, and one of the major controversies in the field relates to reliable SC markers. This study was carried out to evaluate and compare the expression of the stem cell associated marker: ABCG2, keratinocyte stem cell marker: p63 and corneal differentiation markers: Cnx43 and K3/K12 on limbal explants cultured on human amniotic membrane (HAM) with intact epithelium and HAM denuded of its epithelium., Methods: Human limbal biopsies obtained from the cadaveric donor eyes were used in this study. The cells were cultured over the HAM with intact and denuded epithelium. Reverse transcriptase PCR, immunohistochemistry, Western blotting for ABCG2, P63, Cnx43 and K3/K12 were done., Results: The limbal epithelial cells cultured over intact HAM expressed the stem cell associated markers (ABCG2, p63) and showed reduced expression of the differentiation markers (Cnx43 and K3/K12) when compared to limbal epithelial cells cultured over denuded HAM, which expressed more differentiation markers at the end of three weeks. BrdU label retaining cells were observed in the limbal epithelial cells cultured over HAM with epihelium only., Interpretation & Conclusions: Our results showed that the intact HAM supported the growth of limbal epithelial cells expressing stem cell associated markers, and allowing little differentiation of the limbal cells to cornea phenotype. Further studies are needed to understand the properties of the amniotic epithelium that retains the stemness in the cultured limbal stem cells.

Background & Objectives: Cultivated limbal stem cell transplantation is being used as a current treatment modality for limbal stem cell deficiency. However, use of allogenic biological material as substrate is associated with risks of transmission of certain diseases and allograft rejection. Therefore development of non-toxic biodegradable synthetic polymers is important. We undertook this study to evaluate the use of a synthetic polymer Mebiol gel as a substrate for the growth of limbal phenotype cells and cornea phenotype cells from limbal explants., Methods: Human cadaveric limbal explants cells were cultivated on Mebiol gel. The proliferative capacity of cultivated cells was analyzed with thymidine incorporation studies. Immunostaining for presumed limbal stem cell association markers and cornea differentiation markers was performed and confirmed with reverse transcription (RT-PCR)., Results: The limbal explants underwent proliferation in vitro. The cultivated cells expressed the presumed limbal stem cell association markers (ABCG2 and p63), the transient amplifying cell markers (connexin 43, integrin alpha9) and the cornea differentiation marker (K3). RT PCR confirmed the immunohistochemical data., Interpretation & Conclusion: Our findings showed that the synthetic polymer Mebiol gel was able to support limbal explant proliferation. The cultured cells expressed presumed limbal stem cell association markers, transient amplifying cells and cornea phenotype markers. Mebiol Gel can be used as a scaffold for growing limbal explants.

A 27-year-old man had excimer photoastigmatic keratectomy in the right eye and laser in situ keratomileusis in the left eye for the treatment of equivalent myopia. Preoperative slitlamp examination did not reveal evidence of keratoconus, central corneal pachymetry was 485 microm in the right eye and 500 microm in the left eye, and corneal topography revealed asymmetric bow-tie astigmatism with inferior steepening in the right eye and a small area of inferior steepening in the left eye. Twenty-two months after surgery, the patient complained of poor vision in the left eye. Slitlamp examination of the left eye revealed central corneal thinning and protrusion, with a Fleischer ring within the flap. Corneal topographic evaluation revealed a stable map in the right eye and central corneal steepening indicative of keratectasia in the left eye.

Purpose: To report results of smaller diameter corneal flap of original thickness in the management of partial flap during laser in situ keratomileusis (LASIK)., Methods: Retrospective review of the case records of three patients (3 eyes) who had a partial corneal flap during LASIK., Results: Retreatment was performed using an 8.5-mm-diameter corneal flap, which was smaller than the original partial flap of 9.5-mm diameter. The recut depth was maintained as the original cut depth of 160 microm. No intraoperative or postoperative complications were noted. At follow-up 4 weeks later, two patients had an uncorrected visual acuity of 20/20. One patient had uncorrected visual acuity of 20/40 that improved to preoperative best spectacle-corrected visual acuity of 20/30 with a correction of +0.50 -1.25 x 170 degrees., Conclusion: A smaller diameter corneal flap of original thickness can be used for retreatment of partial flap during LASIK. A thicker posterior stromal bed after laser ablation may be retained with this technique, compared to retreatment using a corneal flap of greater thickness.

Objective: To describe a systematic approach to corneal autografting., Patients and Methods: Retrospective review of case records of 3 patients who underwent this procedure., Results: All patients achieved satisfactory visual recovery. Oversized grafts of 0.5 mm had less post-operative complications in the recipient eye. A phakic blind eye that received a 0.5 mm undersized graft had a temporary ocular hypertensive phase., Conclusions: Corneal autografting is a safe and effective technique in selected patients. The preferred surgical technique consists of surgery first on the blind eye, excision of a 0.5 mm oversized donor graft, cataract extraction if the eye is phakic, and use of an appropriate temporary keratoprosthesis to maintain donor eye integrity until autografting is completed in the eye with visual potential.

Purpose: To compare outcomes of limbal autograft transplantation (LAT) in the acute and chronic phases of ocular surface burns., Methods: Retrospective analysis of case records of 16 consecutive patients who underwent LAT for ocular surface burns, at our institute, between April 1994 and March 1997., Results: Limbal autograft transplantation was successful in reconstructing the corneal surface and restoring ocular comfort in 15 (93.8%) eyes. Limbal autografting failed to reconstruct the ocular surface in one patient undergoing surgery 2 weeks after grade IV alkali burns. In 13 eyes with counting fingers or worse vision, functional success (visual acuity >20/400) was attained after LAT in nine (69.2%) eyes. Visual acuity > or = 20/80 was achieved in two (25%) of eight eyes undergoing surgery for a persistent epithelial defect (PED) and five of six (83.3%) eyes undergoing surgery after the epithelial defect had healed (p = 0.03). Nine patients underwent simultaneous superior and inferior limbal autografting. Mean epithelial healing time in six of these patients undergoing surgery in the acute phase of injury (<4 months) was 15+/-6.1 days. In three patients undergoing a similar procedure in the chronic phase of injury, the healing time was 8.3+/-6.7 days., Conclusions: Limbal autograft transplantation is successful in reconstructing the corneal surface and restoring ocular comfort after ocular surface burns. Surgery in the acute phase of injury (<4 months), in the presence of a PED, could result in delayed corneal reepithelialization and poorer visual prognosis. If performed in the acute phase of injury, LAT should be performed after adequate limbal vascularization and resolution of surface inflammation in the recipient eye, avoiding graft placement over ischemic limbus.

Background and Objective: Excimer laser photorefractive keratectomy (PRK) is quickly establishing itself as a dependable treatment modality for the correction of myopia. We review in this report our experience with the Summit OmniMed UV200 excimer laser (Summit Technology, Inc., Waltham, MA) in the treatment of low to high degrees of myopia., Patients and Methods: We retrospectively analyzed the results of excimer laser PRK in 139 eyes of 127 patients treated between November 1993 and August 1994., Results: The study population was comprised of 75 males and 52 females with a mean ago of 26.9 +/- 6.21 years (range, 19 to 52 years). All patients were followed for at least six months and the mean follow-up was 11.7 +/- 3.63 months (range, 6 to 22 months). Mean pre-operative myopia was -8.23 +/- 3.32 diopters (D) (-2.63 to -19.50 D). Of 139 eyes, 101 (72.7%) had a pre-operative myopia greater than -6.00 D; and 32 (23.0%) had a pre-operative myopia greater than -10.00 D. At last follow-up, 73 (52.5%) eyes were within +/-1.00 D of emmetropia, 92 (66.2%) eyes had an uncorrected visual acuity of > or = 6/12, and 12 (8.6%) eyes lost 2 or more lines of spectacle corrected visual acuity. Severe corneal haze (grade 3 to 4) was seen in 20 (14.4%) eyes at last follow-up. Complications encountered in this series included steroid response in 15 (10.8%) eyes, and reactivation of herpes simplex keratitis in one (0.7%) eye. Statistical analysis reveals that the only risk factor independently associated with regression > or = 2.00 D following PRK is the occurrence of severe corneal haze (grade 3 and 4)., Conclusion: Excimer laser PRK appears to be a safe and predictable method for the treatment of myopia < 6.00 D. With higher degrees of myopia the results appeared to be more guarded, mainly due to the healing pattern of the cornea following single zone myopic ablations.

We evaluated the chlamydial direct diagnostic tests and McCoy cell culture methods for detection of Chlamydia trachomatis in 234 conjunctival scrapings from 127 patients. Direct (rapid) diagnostic methods of Giemsa stain cytology, fluorescent antibody test (FAT) and immunoperoxidase (IPO) stain and McCoy cell culture isolation method using IPO staining and FAT for detection of C. trachomatis antigen were carried out. McCoy cell culture isolation using FAT antigen detection method was considered as the gold standard. The specificity of all the direct diagnostic tests were above 90 per cent but sensitivity of Giemsa cytology, FAT and IPO stain were 36, 54 and 53 per cent respectively showing that as direct diagnostic tests Giemsa stain cytology was inefficient and FAT and IPO were useful. Specificity and sensitivity of McCoy cell culture using IPO stain for detection C. trachomatis antigen were above 90 per cent and it is suggested that this test can be used as a routine diagnostic method.