Patient satisfaction and self-reported dry eye symptoms in hyperopic patients treated with femtosecond laser in situ keratomileusis.

PURPOSE: To study self-reported patient satisfaction and dry eye symptoms in hyperopic correction with femtosecond laser-assisted in situ keratomileusis (FS-LASIK). PATIENTS AND METHODS: Ninety-eight eyes (53 patients) were treated with FS-LASIK for hyperopia. Patients' self-reported dry eye symptoms and satisfaction with near and far vision were graded on the visual analog scale (VAS) preoperatively and 1 month postoperatively. RESULTS: Ninety-one percent of the eyes with the plano target (54 eyes) achieved an uncorrected distance visual acuity of 20/20 or better. Predictability, defined as spherical equivalent refraction within ±0.5 D of target, was 88% of all eyes. None of the eyes lost two or more Snellen lines of corrected distance visual acuity. There was no significant change in the self-reported dry eye sensation (VAS score from 2.7±2.0 to 2.8±2.0; P=0.66). In 44 monovision patients, satisfaction with both far vision (from 71.2±19.8 to 89.2±8.7; P<0.0001) and near vision (from 51.7±26.2 to 89.3±13.2; P<0.0001) increased significantly. In nine emmetropic patients, satisfaction with neither far vision nor near vision was significantly improved, although there was a clear tendency (from 73.7±23.7 to 86.9±15.3; P=0.22, and from 58.9±29.1 to 81.6±17.4; P=0.11, respectively). In the monovision patient group, far vision satisfaction decreased when dry eye symptoms increased. Monovision patients, as predicted, were more satisfied with their near vision, when postoperative spherical equivalent from target was on the myopic side. CONCLUSION: FS-LASIK correction of hyperopia significantly improved patient satisfaction with both near and far vision in monovision patients. Hyperopic patients had no significant changes in postoperative dry eye symptoms compared to preoperative values.

Uncorrected visual acuity, postoperative astigmatism, and dry eye symptoms are major determinants of patient satisfaction: a comparative, real-life study of femtosecond laser in situ keratomileusis and small incision lenticule extraction for myopia.

PURPOSE: To compare factors affecting patient satisfaction after femtosecond laser in situ keratomileusis (FS-LASIK) and small incision lenticule extraction (SMILE) for myopia in the real-life situation study. METHODS: The SMILE group included 100 eyes (51 patients) and the FS-LASIK group 200 eyes (102 patients). In addition to clinical examination, dry eye symptoms and patient satisfaction with far and near vision were reported and graded on the visual analog scale preoperatively and one month after the operation. Case-control pairs were selected for the SMILE patients from FS-LASIK-treated patients to ensure the homogeneity in spherical equivalent refraction, preoperative dry eye, and visual satisfaction. RESULTS: Eighty percent of SMILE eyes and 83% of FS-LASIK eyes achieved an uncorrected distance visual acuity of 20/20 or better. Predictability (±0.5 D of mean target spherical equivalent refraction) was 91% in SMILE and 93.5% in FS-LASIK. No eyes lost two or more Snellen lines of corrected distance visual acuity. Based on case-control pairs, dry eye symptoms remained the same after one month in the FS-LASIK-treated eyes (P=0.87) but decreased in the SMILE-treated eyes (P=0.01) compared with the preoperative situation. Patient satisfaction with far vision improved significantly in both groups (P<0.001), but satisfaction with near vision improved significantly only in FS-LASIK (P<0.001) and not in SMILE (P=0.58). There was more postoperative astigmatism in SMILE in comparison with FS-LASIK (P=0.002). CONCLUSIONS: In a real-life situation, patients with preoperative dry eye experience were often directed to the SMILE operation, which resulted in beneficial decrease in their dry eye symptoms. Patient satisfaction with far vision decreased with increasing dry eye symptoms and postoperative astigmatism in both SMILE- and FS-LASIK-treated emmetropic patients. Safety, efficacy, and predictability were comparable in both treatments.

Femtosecond lasers for laser in situ keratomileusis: a systematic review and meta-analysis.

PURPOSE: The aim of this study was to review and meta-analyze whether there are differences between reported femtosecond (FS) lasers for laser-assisted in situ keratomileusis (LASIK) in terms of efficacy, predictability, and safety as primary outcomes and corneal flap thickness measurements and pre- and postoperative complications as secondary outcomes. METHODS: A comprehensive literature search of PubMed, Science Direct, Scopus, and Cochrane CENTRAL Trials Library databases was conducted to identify the relevant prospective randomized controlled trials of FS lasers for LASIK. Thirty-one articles describing a total of 5,404 eyes were included. RESULTS: Based on efficacy, IntraLase FS 10 and 30 kHz gave the best results. Based on predictability and safety, there were no differences between various FS lasers. FEMTO LDV and IntraLase FS 60 kHz produced the most accurate flap thicknesses. IntraLase and Wavelight SF200 had the fewest intraoperative complications. IntraLase, Visumax, and Wavelight FS200 had the most seldom postoperative complications. CONCLUSION: There were dissimilarities between different FS lasers based on efficacy and intraoperative and postoperative complications. All FS lasers were predictable and safe for making corneal flaps in LASIK.

Lasers in corneal surgery.

New laser methods have introduced new possibilities and partly replaced traditional methods in corneal surgery. Femtosecond lasers have traditionally been used to replace the surgeons' knife in corneal refractive surgery and corneal transplantation. Excimer laser has been used from the beginning of 1990 for the reshaping of cornea. With the development of excimer laser, traditional photorefractive corneal surgery has improved and new methods have been found such as PRK, PTK, epi-LASIK, and LASEK. Today LASIK, the most commonly used refractive surgery, uses both femtosecond and excimer lasers. SMILE, the most recent method in corneal refractive surgery, is solely based on the use of femtosecond laser.

Laser-assisted in situ keratomileusis flap creation with the three-dimensional, transportable Ziemer FEMTO LDV model Z6 I femtosecond laser.

PURPOSE: To present the outcomes of laser-assisted in situ keratomileusis (LASIK) operations performed with the new three-dimensional, transportable FEMTO LDV Z6 I femtosecond laser (Ziemer Ophthalmic Systems, Port, Switzerland) and the Allegretto Wave Concerto 500 Hz excimer laser (Wavelight AG, Erlangen, Germany) in terms of accuracy, reproducibility and safety of flap creation. METHODS: This is a retrospective study of 309 consecutive eyes of 160 patients treated with the FEMTO LDV Z6 I for corneal flap creation. The target flap thickness was 90 µm. The size of the suction ring varied from 9.5 to 10.0 mm and target flap diameter from 9.3 to 9.6 mm, respectively. The target hinge length was 4.0 mm. RESULTS: The FEMTO LDV Z6 I produced the 90-µm targeted flaps very consistently (mean 90.1 ± 2.7 µm, range 78-100). Mean flap diameter with the 9.3-mm target flap diameter was 9.3 ± 0.1 mm (range 9.0-9.6) and with the 9.6-mm target flap diameter 9.6 ± 0.1 mm (range 9.0-9.8). Mean hinge length was 3.9 ± 0.1 mm (range 3.3-4.2). Minor complications were reported in 15 (5%) eyes, but none of them prevented refractive laser treatment. The most common complications were bubbles in the conjunctiva (n = 7, 2%) and an opaque bubble layer inside the flap margin (n = 6, 2%). None of the eyes lost two Snellen lines of corrected distance visual acuity during 1-month follow-up. CONCLUSIONS: In the hands of an experienced surgeon, the transfer from the Classic FEMTO LDV to Z6 I was a safe and straight forward process yielding accurate and reproducible flaps.

Laser in situ keratomileusis enhancements with the Ziemer FEMTO LDV femtosecond laser following previous LASIK treatments.

BACKGROUND: The aim of this paper is to present the accuracy, predictability, and safety outcomes of LASIK enhancements performed with the FEMTO LDV femtosecond laser (Ziemer Ophthalmic Systems, Port, Switzerland) and the Allegretto Wave Concerto 500 Hz excimer laser (Wavelight AG, Erlangen, Germany), following previous LASIK treatments. METHODS: FEMTO LDV was used for flap creation in 85 previously LASIK-treated eyes of 62 patients. The intended flap thickness was 90 µm in 81 eyes and 140 µm in 4 eyes. The size of the suction ring was 9.0 mm in 72 eyes and 9.5 mm in 13 eyes. Flap dimensions were measured and correlated to preoperative characteristics. RESULTS: With the intended flap thickness of 90 µm in previously LASIK-treated eyes, the actual flap thickness was 90.2 ± 6.6 µm (range 80-122), and the flap diameter was 9.2 ± 0.2 mm (range 8.7-9.9). The mean hinge length was 4.0 ± 0.2 mm (range 3.0-4.8). Flap thickness correlated positively with patient age and hinge length. Complications were reported in 12 eyes (14.1 %). Most of the complications were very mild, and none of them prevented further refractive laser treatment. One eye lost two Snellen lines of best spectacle-corrected visual acuity. CONCLUSIONS: Femtosecond LASIK enhancement is warranted only in rare cases. Surgical experience is needed and special caution must be practiced. For cases of a primary free cap, femtosecond LASIK is not recommended.

The cytotoxic effects of preserved and preservative-free prostaglandin analogs on human corneal and conjunctival epithelium in vitro and the distribution of benzalkonium chloride homologs in ocular surface tissues in vivo.

PURPOSE: To investigate the cytotoxicity of benzalkonium chloride (BAC)-containing ophthalmic solutions of prostaglandin analogs (latanoprost, travoprost, bimatoprost, and preservative-free (PF) tafluprost), BAC mixture (BACmix) and BAC homologs with different alkyl chain lengths using human corneal epithelial (HCE) and conjunctival epithelial (IOBA-NHC) cell cultures. The distribution of BAC homologs in rabbit ocular surface tissues in vivo was examined. METHODS: The cells were exposed for one hour to prostaglandin analogs, BACmix and three homologs. Cytotoxicity was assessed with the WST-1 and lactate dehydrogenase (LDH) assays for cellular viability and cell membrane integrity. BAC 0.02% solution was instilled on the rabbit eye daily for 14 days and the concentrations of BAC homologs in external ocular tissues were determined. RESULTS: The order of decreasing cytotoxicity in the WST-1 test was latanoprost ≥ travoprost > bimatoprost ≥ PF tafluprost. IOBA-NHC cells were more sensitive than HCE cells. In HCE, only latanoprost diluted to 10% increased LDH leakage. In IOBA-NHC, LDH leakage was statistically significant with 3-10% travoprost and 10% latanoprost. The order of decreasing cytotoxicity of preservatives was C14 > C12 > BACmix > C16 in HCE and C12 > C14 > BACmix > C16 in IOBA-NHC. Following treatment with BAC 0.02% solution, the amounts of BAC-C12, -C14 and -C16 in rabbit cornea and conjunctiva, respectively were: 0.37 ± 0.08 and 2.64 ± 0.27 ng/mg; 0.42 ± 0.07 and 4.77 ± 0.43 ng/mg; 0.04 ± 0.01 and 0.54 ± 0.05 ng/mg. CONCLUSIONS: The cytotoxic effects of latanoprost, travoprost, and bimatoprost were dependent on the BAC concentration in their formulations. BACmix was cytotoxic at the concentrations above those corresponding to 0.001% BAC in ophthalmic medications. PF tafluprost was the least toxic of the drugs tested. Within studied BAC homologs, those with longer alkyl chain and higher lipophility penetrated effectively into rabbit external ocular tissues.

LASIK flap creation with the Ziemer femtosecond laser in 787 consecutive eyes.

PURPOSE: To present the flap characteristics and short-term efficacy and safety of 787 consecutive LASIK procedures with the FEMTO LDV femtosecond laser (Ziemer Ophthalmic Systems) for the treatment of refractive errors. METHODS: Seven hundred eighty-seven consecutive eyes of 405 previously non-operated patients were treated with the FEMTO LDV. Intended flap thickness was 110 microm and intended flap diameter varied from 8.5 to 9.5 mm. Refractive treatment was performed with the WaveLight ALLEGRETTO WAVE Concerto 500 Hz excimer laser. All eyes were wavefront-optimized. RESULTS: The mean flap thickness, measured by ultrasound pachymetry, was 90.0+/-5.5 microm (range: 67 to 107 microm) in right eyes and 90.1+/-4.6 microm (range: 77 to 106 microm) in left eyes. Mean flap diameter was 9.1+/-0.2 mm (range: 8.4 to 9.9 mm) in right eyes and 9.1+/-0.2 mm (range: 8.0 to 10.0 mm) in left eyes. Increasing flap thickness was correlated with increasing corneal thickness in right eyes and flatter keratometric value K(1) in left eyes. The most common complication was minor bleeding during the procedure (12.7%). All other complications were rare (8.4%), and none prevented further laser ablation. CONCLUSIONS: The Ziemer FEMTO LDV laser created thinner LASIK flaps than intended but with a low standard deviation and minimal intraoperative complications.

Comparison of laser in situ keratomileusis reoperation outcomes with the Moria M2 head 90 and 130 following previous photorefractive keratectomy or laser in situ keratomileusis.

PURPOSE: To compare the Moria Model Two (M2) automated microkeratome with the head 90 (intended to create a 120-microm flap) to the head 130 (intended to create a 160-microm flap) in reoperations following previous photorefractive keratectomy (PRK) or laser-assisted in situ keratomileusis (LASIK) in terms of accuracy, predictability, safety and complications of the procedure. METHODS: Eighty-five eyes of 70 consecutive patients received LASIK with the Moria M2 microkeratome. Nine previously PRK-operated eyes were reoperated with the head 90 and 37 eyes were reoperated with the head 130. Repeated LASIK was performed on 16 eyes with the head 90 and on 23 eyes with head the 130. Flap dimensions were measured and correlated to preoperative parameters. RESULTS: The average flap thickness in the previously PRK-operated eyes was 115.1 microm [range 82-137 microm, standard deviation (SD) 17.9] with the head 90 and 131.2 microm (range 105-171 microm, SD 19.8) with the head 130. In the previously LASIK-operated eyes, the mean flap thickness was 139.2 microm (range 92-182 microm, SD 23.8) with the head 90 and 141.9 microm (range 109-179 microm, SD 15.2) with the head 130. There were no free or incomplete flaps or flaps with buttonholes in the study. There was no statistically significant difference in postoperative uncorrected visual acuity (UCVA) between the groups. CONCLUSION: In eyes with previous PRK or LASIK, LASIK reoperation offers a safe alternative for improving refractive outcomes. The Moria M2 head 90, especially in LASIK-operated eyes, does not cut thinner flaps compared to the head 130.

Corneal flap thickness with the Moria M2 microkeratome and Med-Logics calibrated LASIK blades.

PURPOSE: This study aimed to compare and study potential factors that affect the accuracy of corneal flap thickness created in laser-assisted in situ keratomileusis (LASIK) using the Moria model 2 (M2) head 130 microkeratome with the Med-Logics calibrated LASIK blades Minus 20 (ML -20) and Minus 30 (ML -30). METHODS: Corneal thickness in 200 (164 myopic and 36 hyperopic) eyes (100 patients) was measured by ultrasonic pachymetry preoperatively and intraoperatively after flap cutting. A total of 100 eyes were treated with the ML -20 and 100 with the ML -30. The right eye was operated before the left eye in each patient, using the same blade. In an additional group of 40 eyes, the left eye was operated first. RESULTS: Mean corneal flap thickness using the ML -20 blade for an intended flap thickness of 140 µm was 129.1 µm (standard deviation [SD] 15.6, range 104-165 µm) in right eyes and 111.5 µm (SD 14.5, range 78-144 µm) in left eyes. Mean corneal flap thickness using the ML -30 blade for an intended flap thickness of 130 µm was 127.1 µm (SD 16.6, range 90-168 µm) in right eyes and 109.9 µm (SD 16.8, range 72-149 µm) in left eyes. CONCLUSIONS: Both microkeratome blade types cut thinner flaps than were intended. There was substantial variation in flap thickness. The first flap to be cut with a particular blade was considerably thicker than the second flap cut with the same blade. Based on these data, we recommend the use of disposable single-use microkeratomes rather than these ML blades.

The effects of 5-fluorouracil on ocular tissues in vitro and in vivo after controlled release from a multifunctional implant.

PURPOSE: To evaluate the effects of 5-fluorouracil (5-FU) on ocular cells in vitro and the effects of degradable 5-FU-loaded poly(DL-lactide-co-glycolide; PDLGA) 50:50 implant in the rabbit eye in vivo. METHODS: Cytotoxicity was assessed with a tetrazolium salt WST-1 cell proliferation/viability test and a lactate dehydrogenase (LDH) leakage test in rabbit corneal stromal fibroblasts (SIRCs), bovine corneal endothelial cells (BCECs), human conjunctival epithelial cells (IOBA-NHCs), human retinal pigment epithelial cells (ARPE-19), and human corneal epithelial cells (HCECs). The 5-FU-loaded PDLGA implants were surgically placed in rabbit eyes with a deep sclerectomy technique and the histopathology of the eyes was examined. RESULTS: In vitro, 5-FU affected cell proliferation and survival in a time- and dose-dependent manner. In the WST-1 test, adverse effects in serum-free conditions started from 0.0005 mg/mL 5-FU in SIRCS and HCECs, whereas in other cell types, 0.005 mg/mL 5-FU hindered cell proliferation. In serum-free conditions 72-hour 5 mg/mL 5-FU treatment decreased cell viability to 40% in BCECs and to 10% to 15% in other cell types. 5-FU had no or very minor effects on LDH leakage. In vivo, the 5-FU implant showed no signs of toxicity in cornea and retina, whereas in the conjunctival stroma near the implantation site, some inflammatory cells and a marked subepithelial condensation of stromal connective tissue was observed during the postoperative period of 4 weeks. CONCLUSIONS: 5-FU had a broad therapeutic range, and the 5-FU implant showed only minor tissue reactions in conjunctiva at the surgical site. 5-FU is a possible candidate for controlled drug release.

In vitro biocompatibility of degradable biopolymers in cell line cultures from various ocular tissues: extraction studies.

In vitro biocompatibility of 50:50 PDLGA, 85:15 PDLGA, and Inion GTR(TM) membrane was evaluated in cell line cultures from various ocular tissues, in human corneal epithelial cells (HCE), rabbit stromal fibroblasts (SIRC), bovine corneal endothelial cells (BCE), human conjunctival epithelial cells (IOBA-NHC), and human retinal pigment epithelial cells (ARPE-19). To study the toxicity of degradation products, the biomaterials were extracted in phosphate buffered saline at 70 degrees C for 24 h. The cell cultures were exposed to biomaterial extract diluted in medium (1:1-1:8) and the biocompatibility was evaluated by the WST-1 cytotoxicity/cell proliferation test. In all experiments without pH neutralization, cell viability increased with decreasing biomaterial extract volume. The highest extraction ratio 1:1 of PDLGA 50:50 decreased viability from 5-20%, from the control level, depending on the cell type. The corresponding cell viability values for PDLGA 85:15 and Inion GTR membrane ranged from 47-87% and 66-92%, respectively. When the pH of biomaterial extract was neutralized, Inion GTR membrane and PDLGA 85:15 had no effect on viability. BCE, HCE, and IOBA-NHC appeared to the most sensitive cell types, while SIRC and ARPE-19 were more resistant. The results of our in vitro studies suggest that the polymers tested are satisfactorily biocompatible.

Corneal flap thickness with the Moria M2 single-use head 90 microkeratome.

PURPOSE: To analyse the accuracy of corneal flap thickness created in laser-assisted in situ keratomileusis (LASIK) using the Moria Model 2 (M2) single-use head 90 microkeratome. METHODS: The corneal thickness of 300 (266 myopic and 34 hyperopic) eyes of 150 patients was measured by ultrasonic pachymetry preoperationally and intraoperationally after flap cut. The Moria M2 single-use head 90, intended to create a flap with a thickness of 120 microm, was used in all eyes. The right eye was always operated first and the left eye second, using the same blade. RESULTS: Mean corneal flap thickness was 115.4 microm (standard deviation [SD] 12.5) in the two eyes, 115.7 microm (SD 12.4, range 73-147 microm) in right eyes and 115.1 microm (SD 12.6, range 74-144 microm) in left eyes. Mean horizontal flap diameter was 9.1 mm (SD 0.2) and mean hinge length 4.1 mm (SD 0.1). There were no free flaps, incomplete flaps or flaps with buttonholes in the study. Occasional iron particles were observed in three (1.0%) eyes. CONCLUSIONS: As with most microkeratomes, the single-use head 90 microkeratome cut thinner flaps than were intended. The range of the cuts was relatively wide. However, thin flaps did not increase the rate of flap-related complications. The difference between the first and second eyes was not significant.

Bilateral comparison of corneal flap dimensions with the Moria M2 reusable head and single use head microkeratomes.

PURPOSE: To compare the Moria (Antony, France) M2 automated microkeratome with the head 130 to a new disposable single use head to evaluate complications, accuracy, and safety of the procedure. METHODS: Ninety-eight eyes of 49 consecutive patients were operated with the Moria M2 microkeratome. One eye was operated with the metallic head 130 and the other with a plastic single use head, both designed to create a 160-microm flap. Intraoperative flap dimensions were correlated to preoperative parameters and evaluated 1 month postoperatively. RESULTS: With the head 130, mean thickness was 153.3 microm (standard deviation [SD] 13.3, range: 102 to 179 microm). When using a single use head, mean thickness was 148.0 microm (SD 9.8, range: 120 to 170 microm). Occasional iron particles were observed in one eye with both head types. No true epithelial ingrowth was detected in any of the eyes, but epithelial dots at the wound edge occurred in one eye, when using the head 130, but not in the eyes operated with a single use head. CONCLUSIONS: On average, both head types created thinner flaps than attempted. Single use heads produced thinner flaps than the head 130. Accuracy in flap thickness in terms of standard deviation was significantly better in single use heads than in the head 130. Single use heads also had fewer microkeratome-related complications. In clinical practice, the single use head was easier to use because no assembly was required. Plastic single use heads also worked more smoothly than the metallic head 130.

Corneal flap measurements in laser in situ keratomileusis using the Moria M2 automated microkeratome.

PURPOSE: To evaluate accuracy and predictability and factors that influence the dimensions of the laser in situ keratomileusis (LASIK) corneal flap created with the Moria M2 automated microkeratome (Moria SA, Antony, France). METHODS: The flap thickness of 454 eyes of 243 consecutive patients was measured using subtraction ultrasonic pachymetry during LASIK with the Moria M2 microkeratome head 130 designed to create a 160-microm-thick flap. Flap dimensions were evaluated and measurements were correlated with preoperative parameters. A stepwise regression analysis was used to determine the factors that influenced actual flap thickness. RESULTS: The preoperative spherical equivalent refraction of the 454 eyes ranged from -12.125 diopters (D) to +6.25 D. Patient age ranged from 18 to 57 years (mean age: 31.3 +/- 8.8 years). Mean preoperative keratometric power K1 was 44.31 +/- 1.59 D and K2 was 43.32 +/- 1.54 D. Mean preoperative central comeal thickness was 552.4 +/- 32.5 microm (range: 466 to 665 microm). With an attempted thickness of 160 microm, the Moria M2 flap thickness ranged from 77 to 209 microm (mean: 153.3 +/- 19.0 microm). Mean horizontal flap diameter was 9.2 +/- 0.2 mm and mean hinge length 4.6 +/- 0.3 mm. Increasing flap thickness was found to correlate with increasing preoperative comeal thickness, younger patient age, and flatter preoperative keratometric power K1. CONCLUSIONS: Although the standard deviation of the flap thickness was relatively small, remarkable individual variation was noted. Therefore, the intraoperative calculation of the remaining stromal bed is recommended. Furthermore, the consideration of central corneal thickness, patient age, and preoperative keratometry are helpful parameters to avoid too deep ablation.

Protein response of insect cells to bioreactor environmental stresses.

Protein expression of Spodoptera frugiperda (Sf9) insect cells was characterized upon exposure to environmental stresses typically present in bioreactors including heat shock, oxygen deprivation, shear stress, change of pH, and salinity or ethanol shock. This study fills the void in knowledge as to how bioreactor hydrodynamics, anoxia, small changes in pH as well as salinity alterations due to pH control or exposure to ethanol used in asepsis treatments affect protein expression in Sf9 cells. Heat shock at 43 degrees C induced proteins at 83 kDa, 68-78 kDa and six small heat shock proteins (hsps) at 23-15.5 kDa. Anaerobic conditions in CO2 atmosphere reduced significantly the normal protein synthesis and induced a small subset of heat shock proteins at 70 kDa. Oxygen deprivation in nitrogen atmosphere transiently induces the 70 kDa proteins and had minor effects on the normal protein synthesis. Exposure to increased salinity or ethanol concentration failed to trigger the stress response, but may extensively inhibit the induction of normal proteins even though there was a negligible change in cell viability. Shear stress that had a major reducing effect on cell viability did not change the protein synthesis profile of Sf9 cells. Both long and short term exposures to small pH changes had negligible effects on protein synthesis.

Geldanamycin increases 4-hydroxynonenal (HNE)-induced cell death in human retinal pigment epithelial cells.

Development of age-related macular degeneration (AMD) is associated with functional abnormalities and cell death in retinal pigment epithelial (RPE) cells attributable to oxidative stress. To minimize the adverse effects of oxidative stress, cells activate their defence systems, e.g., via increased expression of heat shock protein (Hsp), activation of stress sensitive AP-1 and NF-kappaB transcription factors. In this study, we examined the accumulation of Hsp70 protein, activation of AP-1 and NF-kappaB transcription factors in human ARPE-19 cells subjected to a 4-hydroxynonenal (HNE)-induced oxidative stress. In addition, the influence of Hsp90 inhibitor geldanamycin (GA) was studied in HNE-treated cells. Mitochondrial metabolic activity and apoptosis were determined to evaluate cell death in the ARPE-19 cells. The ARPE-19 cells showed increased accumulation of Hsp70 protein before of the cytotoxic hallmarks appearing in response to HNE. In contrast, increased DNA-binding activities of AP-1 or NF-kappaB transcription factors were not seen under HNE insults. Interestingly, GA significantly increased cell death in the HNE-treated cells, which was involved in caspase-3 independent apoptosis. This study reveals that the Hsps have an important role in the cytoprotection of RPE cells subjected to HNE-derived oxidative stress.

Eight-year follow-up of photorefractive keratectomy for myopia.

PURPOSE: We evaluated 8-year results of excimer laser photorefractive keratectomy (PRK) for myopia in terms of stability and late complications. METHODS: Ninety-two myopic eyes of 55 patients were treated with a single-step method using an Aesculap-Meditec MEL 60 excimer laser with a 5.0-mm ablation zone. Treated eyes were divided into three groups according to preoperative refraction: low myopes (< or = -6.00 D), medium myopes (-6.10 to -10.00 D), and high myopes (>-10.00 D). RESULTS: Change in myopic regression stabilized in all myopia groups within 12 months, although a small myopic shift occurred up to 8 years after PRK. Mean change in refraction between 2 and 8 years was -0.42 +/- 0.48 D for low myopes, -0.37 +/- 0.34 D for medium myopes, and -0.41 +/- 0.50 D for high myopes. The percentage of eyes within +/- 1.00 D of emmetropia 8 years after PRK was 78.3% in the low myopia group, 68.8% in the medium myopia group, and 57.1% in the high myopia group. One eye lost 2 lines of best spectacle-corrected visual acuity due to irregular astigmatism. In 13.0% of eyes, a residual trace corneal haze was observed, which had no effect on visual acuity. Apart from the loss of 2 lines of BSCVA in one eye, there were no other late complications during the study period. CONCLUSIONS: The mean change in refraction between 2 and 8 years was less than -0.50 D, regardless of preoperative refraction, and may be attributed to natural age-related refractive change. The appearance of residual corneal haze after 8 years correlated with the amount of myopic correction. PRK was a safe and stable surgical procedure in this group of patients.

A collaborative evaluation of the cytotoxicity of two surfactants by using the human corneal epithelial cell line and the WST-1 test.

This study was undertaken to investigate the use of the in vitro test WST-1, an assay of cell proliferation and viability, for a preliminary safety evaluation of topical ophthalmic preparations. The cytotoxicity of two surfactants, benzalkonium chloride (BAC) and polyoxyethylene-20-stearyl ether (Brij78, PSE) was independently investigated in four laboratories in the EU by using an immortalized human corneal epithelial (HCE) cell line. The HCE cells were exposed to BAC and PSE for 5 min, 15 min, and 1 hour, and the results of the HCE-WST-1 tests were collected and compared. After one-hour exposure, the EC(50) values in BAC-treated cells in the presence of serum ranged between 0.0650 +/- 0.0284 (mean +/- SD) mM, and those in the absence of serum 0.0296 +/- 0.0081 mM. The corresponding values for PSE were 0.0581 +/-.0300 mM and 0.0228 +/-.0063 mM. There were variations in the results between different laboratories, with coefficients of variation ranging from 31 to 121%, mean 58%. The use of one-hour exposure time is to be preferred, and the elimination of serum in the culture medium is recommended to avoid both underestimation of toxic effects and variability of the test results.