Development of singleplex and multiplex real-time (Taqman®) RT-PCR assays for the detection of viruses associated with fig mosaic disease.

Singleplex and multiplex real-time (TaqMan®) RT-PCR assays were developed to detect seven fig-infecting viruses, i.e., fig leaf mottle-associated virus 1 (FLMaV-1), fig leaf mottle-associated virus 2 (FLMaV-2), fig mild mottle-associated virus (FMMaV), fig mosaic virus (FMV), fig latent virus 1 (FLV-1), fig cryptic virus 1 (FCV-1) and fig fleck-associated virus (FFkaV). The sensitivity of the newly developed TaqMan® assays was compared with the corresponding conventional RT-PCR (RT-PCR) using 10° to 10-6 serial dilutions of both cDNA and crude fig extracts. The results showed that the Taqman® RT-PCR assays were generally 102 to 103-fold more sensitive than the RT-PCR assays, except in the case of FLV-1 detection, where the two techniques had the same sensitivity. In the multiplex Taqman® RT-PCR, only a maximum of five viruses could be detected simultaneously in naturally infected fig trees, regardless of which combination of the virus-specific probes and primers were used. Both the RT-PCR and Taqman® RT-PCR assays were used in a large-scale survey of 100 field-grown fig trees in Egypt. The results showed the presence of all seven viruses under study, mostly occurring as mixed infections (63 %). The prevalence of infections observed in the tested samples were as follows: FMV (62 %), FFkaV (59 %), FLMaV-2 (32 %), FLV-1 (16 %), FLMaV-1 (14 %), FCV-1 (7%) and FMMaV (4%). FMV was invariably associated with diseased trees that presented mosaic-like symptoms. In the few cases where the mosaic-affected trees were found to be free of FMV, they were found to be infected with a mixture of two or more other viruses.

Visian implantable contact lens versus AcrySof Cachet phakic intraocular lenses: comparison of aberrmetric profiles.

PURPOSE: To evaluate the in vivo aberrometric performance of two phakic intraocular lenses (PIOLs); the Visian implantable contact lens (ICL) and the AcrySof Cachet PIOL. METHODS: Thirty eight eyes from 29 patients with an age range of 23-32 years and more than 9 D of myopia were divided into two groups; one group of 20 eyes received a Visian ICL model V4c, another group of 18 eyes received an AcrySof Cachet PIOL. Total higher order aberrations (HOAs) root mean square, total coma, and total spherical aberrations were recorded pre and 6 weeks postoperatively to evaluate and compare the aberrometric performance of the Visian ICL and the AcrySof Cachet PIOL implanted in highly myopic patients. RESULTS: Preoperatively, there were no significant differences in any studied parameters, except for preoperative spherical equivalent. Postoperatively, there were no statistically significant differences in the induction of HOAs between both PIOLs. The reduction in spherical aberrations was statistically significant in each group as well as in all study patients. CONCLUSION: Both AcrySof Cachet PIOL and ICLs are effective phakic implants to correct high refractive errors. They both induce small amounts of negative spherical aberration that do not affect the total HOAs, yet reduce the positive ocular spherical aberration. This result is expected to improve the quality of vision in such patients.

Comparison of different intraocular pressure measurement techniques in normal eyes, post surface and post lamellar refractive surgery.

BACKGROUND: The purpose of this study was to determine the accuracy of intraocular pressure (IOP) measurement after laser in situ keratomileusis (LASIK) or epithelial laser in situ keratomileusis (Epi)-LASIK using Goldmann applanation tonometry, air puff tonometry, ocular response analyzer corneal compensated IOP (ORA IOPcc) and Pentacam corrected IOP. METHODS: A prospective comparative clinical study was conducted between February and September 2011 on 30 eyes divided into four groups, i.e. 20 corneas of 10 patients before LASIK (group A), 20 corneas of the same patients 2 months postoperatively (group B), 10 corneas of five patients before Epi-LASIK (group C), and 10 corneas of the same patients 2 months postoperatively (group D). Patient age ranged from 20 to 50 years. IOP was measured using Goldmann applanation and air puff tonometry, ORA corneal compensation, and Pentacam correction (which also measured central corneal thickness). RESULTS: Significant positive linear correlations were found between IOP values measured by Goldmann applanation tonometry and other techniques, and with preoperative pachymetry in group A. The correlation between preoperative Pentacam-corrected and preoperative ORA corneal-compensated IOP was strongest for Goldmann applanation tonometry (r = 0.97 and r = 0.858 respectively, P < 0.001). Compared with preoperative values, postoperative IOP measured by the four methods were significantly lower. The difference was statistically significant when IOP was measured using Goldmann applanation and air puff tonometry compared with the ORA and Pentacam methods (P < 0.001 for LASIK patients and P = 0.017 for Epi-LASIK patients). Nonsignificant correlations were found between the degree of lowering of postoperative IOP and postoperative pachymetry in groups B and D. CONCLUSION: Refractive surgery causes significant lowering of IOP as measured using Goldmann applanation tonometry, air puff tonometry, ORA compensation, and Pentacam correction. LASIK has a greater effect than Epi-LASIK on IOP measurement error following refractive surgery.

Early bubble modification of the big bubble technique for deep anterior lamellar keratoplasty.

PURPOSE: We modified the deep anterior lamellar keratoplasty big bubble technique to improve safety and ease of performance. METHODS: We describe a modification of the big bubble technique that involves injecting the air bubble through a peripheral corneal incision 1 mm from the limbus. The incision is made with a limbal relaxing incision knife. This is done before trephination of the recipient cornea, hence the name "early bubble." The goal of this technique is to reduce the risk of intraoperative corneal perforation and to obtain a large air bubble between Descemet membrane and the corneal stroma. We report outcomes of this technique in 21 eyes. RESULTS: At 6 months, 16 eyes showed a best-corrected visual acuity of 20/50; the remaining eyes had a best-corrected visual acuity of 20/80 or better. The postoperative corneal astigmatism was 3.6+/-0.9 diopters at 3 months and 3.15+/-0.67 diopters at 6 months. The rate of conversion to penetrating keratoplasty was 14%. CONCLUSIONS: We found that the "early bubble" technique improved the ease of performance, safety, and predictability of deep anterior lamellar keratoplasty.

Effect of intracameral bevacizumab injection on corneal endothelial cells: an in vivo evaluation.

BACKGROUND: We studied the effect of an intracameral bevacizumab injection on the corneal endothelium and iris neovessels (INV). METHODS: Ten eyes were studied in 10 patients with an average age of 52.1 +/- 13.17 years. Patients underwent intracameral bevacizumab injections at a concentration of 1.25 mg/0.05 mL. Intraocular pressure, best-corrected visual acuity, gonioscopy, neovascular membrane extent, anterior segment photography, iris fluorescein angiography, pachymetry, and specular microscopy were recorded preoperatively and postoperatively. RESULTS: The minimum follow-up period was 4 months. INVs started to regress within the first 2 days after the injection and regressed completely by the end of the fourth week. Reduction in INV leakage started 1 week after injection, and resolved in 8 eyes (80%) by the end of the fourth week. Intraocular pressure dropped significantly from 17.8 +/- 4.8 mmHg to 16.6 +/- 2.8 mmHg over 4 weeks. The mean endothelial cell loss was 3.95% +/- 6.78%. CONCLUSIONS: An intracameral bevacizumab injection proved to be safe for corneal endothelial cells.

Topography-guided laser in situ keratomileusis (TOPOLINK) to correct irregular astigmatism after previous refractive surgery.

PURPOSE: To assess whether topography-driven laser in situ keratomileusis (LASIK) can correct induced corneal irregular astigmatism. METHODS: A prospective non-comparative case series of 41 eyes (38 patients) with irregular astigmatism following corneal refractive surgery, included two groups: Group 1 (26 eyes) with a defined topographic pattern and Group 2 (15 eyes) with no pattern. Ablation was performed using the Technolas 217C excimer laser with a software ablation program (TOPOLINK) based on corneal topography. Uncorrected and best spectacle-corrected visual acuity, manifest and cycloplegic refraction, corneal topography, superficial corneal surface quality, and image distortion were measured. RESULTS: At 6 months follow-up in Group 1 eyes (defined topographic pattern) mean preoperative BSCVA improved from 0.16 +/- 0.11 LogMAR (0.4 to 0) to 0.09 +/- 0.10 LogMAR (0.2 to 0) (P = .001) (safety index of 1.1). In Group 2 eyes (no pattern), mean preoperative BSCVA was 0.18 +/- 0.11 LogMAR (0.4 to 0), similar to the postoperative BSCVA of 0.17 +/- 0.10 LogMAR (0.3 to 0) (safety index of 0.98). Mean postoperative UCVA was > or = 0.3 LogMAR in 25 eyes (96.2%) in Group 1 (efficacy index of 0.8) and 7 eyes (46.6%) in Group 2 (efficacy index of 0.5). Both superficial corneal surface quality and image distortion significantly improved in Group 1; there were no significant changes in Group 2. We reoperated nine eyes (21.9%). CONCLUSIONS: Topographic-assisted LASIK was helpful in selected cases where irregular astigmatism showed a pattern. It was ineffective in undefined irregular astigmatism. Partial correction of the irregularity and regression of the obtained effect was common.

Artisan phakic iris claw intraocular lens for high primary and secondary hyperopia.

PURPOSE: To evaluate the efficacy, predictability, and safety of the Artisan iris claw phakic intraocular lens for the correction of high primary and secondary hyperopia. METHODS: Fifty-seven eyes were divided into two groups: 29 eyes had primary hyperopia (mean refraction 6.06 +/- 1.26 D, and 28 eyes had secondary hyperopia, (mean refraction 5.88 +/- 1.88 D) induced or residual following a previous corneal refractive procedure. Consecutive implantation of the Artisan iris claw phakic intraocular lens was performed. Main outcome measures recorded were BSCVA, UCVA, refraction, and astigmatic change, intraocular inflammation, and endothelial cell loss. RESULTS: Primary hyperopic group: Preoperatively, mean UCVA was 0.4 +/- 0.7 and mean BSCVA was 0.2 +/- 0.6. After implantation, mean UCVA was 0.3 +/- 0.6 and BSCVA was 0.1 +/- 0.6. Mean cycloplegic residual spherical refractive error after surgery was 0.10 +/- 0.57 D (range -1 to +2 D). Mean surgically induced astigmatism was 1.48 +/- 0.89 D. Safety index was 1.11. Efficacy index was 0.83. Secondary hyperopic group: Preoperatively, mean UCVA was 0.5 +/- 0.7 and mean BSCVA was 0.2 +/- 0.6. Postoperatively, mean UCVA was 0.4 +/- 0.7 and mean BSCVA was 0.2 +/- 0.6. Mean cycloplegic residual spherical refractive error was 0.55 +/- 1.49 D. Mean surgically induced astigmatism was 1.85 +/- 1.19 D. Safety index was 1.05. Efficacy index was 0.7. Postoperative iridocyclitis was observed in one eye (3.4%) in the primary group and in three eyes (10.7%) in the secondary group. Overall corneal endothelial cell loss at 1 year of follow-up was 9.4%. CONCLUSION: The Artisan iris claw phakic intraocular lens was reasonably safe and predictable for correcting high hyperopia.

Phacoemulsification in the anterior chamber.

PURPOSE: To evaluate anterior chamber phacoemulsification as an alternative to an endocapsular procedure. SETTING: Instituto Oftalmológico de Alicante, Alicante, Spain. METHODS: In this prospective randomized masked clinical trial, 60 eyes of 30 patients had cataract extraction by phacoemulsification. A conventional stop-and-chop technique was used in 30 eyes; the other 30 had surgery by the phaco-out technique in which the nucleus was prolapsed into the anterior chamber using extensive hydrodissection. Copious sodium hyaluronate 3.0%-chondroitin sulfate 4.0% (Viscoat) and hydroxypropyl methylcellulose were used to protect the cornea and iris. The phaco time and power, postoperative inflammation, corneal edema (clinically and by pachymetry), and endothelial cell count between groups preoperatively and postoperatively were compared. All patients completed a 3-month follow-up. Postoperative examinations were at 3 days, 2 weeks, and 1 and 3 months. RESULTS: The mean nuclear hardness was 2.67 +/- 0.61 (SD) in the stop-and-chop group and 2.65 +/- 0.71 in the phaco-out group. The power-to-time ratio was 18.47 and 18.80, respectively. The mean endothelial cell loss was 11.18% +/- 4.24% in the stop-and-chop group and 11.20% +/- 5.60% in the phaco-out group. There were no significant differences between groups in any parameter (P >.05, independent-sample t test). CONCLUSION: Phacoemulsification in the anterior chamber was as safe as endocapsular phacoemulsification using a stop-and-chop technique. This technique is fast and easier to learn than endocapsular phacoemulsification.