Levodopa Positively Affects Neovascular Age-Related Macular Degeneration.

BACKGROUND: Age-related macular degeneration (AMD) is a common cause of blindness worldwide. Neovascular AMD (nAMD) is an advanced form of the disease, in which excess vascular endothelial growth factor (VEGF) induces growth of new blood vessels that leak fluid, accounting for 90% of vision loss in AMD. Dysfunction of the retinal pigment epithelium likely initiates AMD. Retinal pigment epithelial cells express a G protein-coupled receptor, GPR143, which downregulates VEGF in response to levodopa. Anti-VEGF therapy effectively treats nAMD, suggesting that excessive VEGF activity drives the pathology. METHODS: In an open-label pilot study, in patients with newly diagnosed nAMD and naïve to anti-VEGF injections (Cohort-1), the effects of carbidopa-levodopa on vision and anatomic outcomes were evaluated for 4 weeks. Then patients were followed 5 months further with ascending levodopa doses. Patients previously treated with anti-VEGF injection therapy (Cohort-2) were also treated with ascending levodopa doses and evaluated for 6 months. RESULTS: Levodopa was safe, well tolerated, and delayed anti-VEGF injection therapy while improving visual outcomes. In the first month, retinal fluid decreased by 29% (P = .02, n = 12) without anti-VEGF treatment. Through 6 months the decrease in retinal fluid was sustained, with a mean frequency of 0.38 injections/month. At month 6, mean visual acuity improved by 4.7 letters in Cohort-1 (P = .004, n = 15) and by 4.8 letters in Cohort-2 (P = .02, n = 11). Additionally, there was a 52% reduction in the need for anti-VEGF injections in Cohort-2 (P = .002). CONCLUSIONS: Our findings suggest efficacy and support the pharmacological targeting of GPR143 with levodopa for the treatment of nAMD in future studies.

Rapid microbial quantification of disinfected contact lens surfaces.

AIM: The efficacy of disinfection of contact lenses is difficult to determine. We have monitored microbial contamination on surfaces and in fluids by intrinsic fluorescence of microbes and distinguished their metabolic states (viable cells, nonviable cells, and endospores). This approach is sensitive (10 cells), requires no added reagents or sample contact, and measurements can be made in near real time. The disinfection performance of AMO Easy Rub™ and Alcon No Rub™ contact lens solutions was compared with CVS-brand saline for contact lenses on contact lenses contaminated with Pseudomonas aeruginosa (ATCC 10145). Our aim is to show that intrinsic fluorescence measurements can yield real-time, critical information about the efficacy of contact lens decontamination products and protocols. MATERIALS AND METHODS: Intrinsic fluorescence of Acuvue™ contact lenses (samples and controls) was measured before and after incubation with P. aeruginosa. Inoculated lenses were then cleaned and disinfected, according to directions for AMO Easy Rub™ and Alcon No Rub™, rubbed and rinsed with saline for contact lenses (the brand made a difference in the experiment), followed by measurement of the effectiveness of the disinfection procedure with the intrinsic fluorescence instrument. Both the sample lenses after disinfection and the control lenses were immediately placed in LB broth for outgrowth and measured by standard cell-counting methods. The experiments were performed in triplicate. RESULTS: The concentration of P. aeruginosa formed on the lenses by the above methods varied from 10(6)-10(13) cells/mL. All lenses that were cultured following cleaning yielded no colonies. However, 10(5)-10(9) cells/mL of the bacteria on the treated lenses remained viable when analyzed by intrinsic fluorescence methodology. CONCLUSIONS: These results indicate that a medically significant amount of bacteria remained on the contact lenses after disinfection and/or cleaning, which were viable but nonculturable and are likely to go undetected when using standard culture methods.

Is tisseel a viable option in posterior lamellar keratoplasty?

PURPOSE: It is well known that Tisseel Fibrin Sealant provides an excellent tissue adhesive. However, its thick and viscuous nature makes it nearly impossible to apply it in a uniform and thin layer. We propose applying the sealant phase as a dry powder and polymerizing it in-vivo by exposing it to thrombin solution after a graft has been placed. METHODS: For each experiment two rabbit corneal buttons were affixed to each other using either Tisseel or thrombin plus dry fibrinogen component, the tensile strength of the bond was then tested in the tension box. Balanced salt solution was used as an aqueous substitute. RESULTS: Tisseel was demonstrated to create significant adhesive tensile force (expressed as N/m) between corneal buttons (P < 10). Using only the dry fibrinogen component followed by injection of the thrombin solution directly into the balanced salt solution did not significantly alter the strength of the bond (P = 0.18). CONCLUSION: The use of the dry fibrinogen component followed by injection of thrombin solution into the balanced salt solution, without the accompanying fibrinolysis inhibitor, is equally effective in adhesive strength when compared to complete Tisseel. This technique may be used in lamellar corneal surgery, although there would be potential difficulties with its application in the in vivo setting.

Prophylactic gatifloxacin therapy in prevention of bacterial keratitis in a rabbit laser in situ keratomileusis model.

PURPOSE: Use the ID(50) (infectious dose to 50% of experimental animals) to quantify the most effective prophylactic dosing regimen to use with gatifloxacin 0.3% (Zymar) for the prevention of keratitis in a rabbit laser in situ keratomileusis model of Staphylococcus epidermidis infection. SETTING: University Laboratory, University of Arizona, Tucson, Arizona, USA. METHODS: Two groups of rabbits were compared in each of 2 experiments that were separated by 12 months. In the first experiment, rabbits receiving no postoperative antibiotic therapy (Group 1) were compared with rabbits receiving postoperative antibiotic therapy (Group 2). In the second experiment, postoperative antibiotic therapy (Group 3) was compared with preoperative and postoperative antibiotic therapy (Group 4). All antibiotic regimens used gatifloxacin 0.3%. Before antibiotic therapy began, corneal pockets were created in the right eye of each rabbit and all rabbits received balanced salt solution (BSS) only or BSS and S epidermidis inoculations in the corneal pocket. Rabbits were monitored for corneal infiltrates after surgery. RESULTS: The ID(50) of the first, second, third, and fourth groups of rabbits was 10(2), 10(4), 10(5), and 10(7) organisms, respectively. The data showed a statistically significant difference between rabbits receiving BSS only and most rabbits receiving BSS plus inoculate at each postoperative measurement (P<.05). CONCLUSIONS: The findings suggest that the use of both preoperative and postoperative antibiotic therapy may be most effective in preventing infection. Postoperative antibiotic therapy increased the number of S epidermidis necessary to cause infection by at least 100-fold over no antibiotic intervention. Preoperative plus postoperative antibiotic therapy increased the number of bacteria necessary to cause infection by at least 100-fold over postoperative therapy alone and by more than 10000-fold over no antibiotic intervention.

Femtosecond laser preparation of donor tissue from the endothelial side.

PURPOSE: The femtosecond laser (Intralase) may provide advantages for dissecting a thin, uniform thickness posterior lamellar disk of donor tissue to be used for endothelial transplantation. We investigated the use of the Intralase to dissect the donor cornea from the posterior side to better obtain a thin and uniform lamellar disk. We investigated the use of a viscoelastic "cushion" to protect the endothelium during applanation and laser delivery. METHODS: Human eye bank donor buttons were placed endothelial side up, covered with a thin coat of viscoelastic, and brought into contact with the Intralase applanation lens. A 7-mm diameter, 100-microm lamellar disk was cut from the endothelial side. The endothelial viability after these procedures was determined using a live cell/dead cell assay. Controls were designed to assess the endothelial viability after applanation and laser application using only a balanced salt solution (BSS) cushion instead of viscoelastic material. Additionally, applanation without lasering using either BSS or a viscoelastic cushion was studied. RESULTS: The average endothelial cell loss in the laser experiment sets were 10% (n = 5, range of 4-17%, Sodium Hyaluronate), 14% (n = 5, range of 7-19%, Sodium Hyaluronate-Sodium Chondroitin) and 6% (n = 5, range of 3-11%, Hydroxypropylmethyl-cellulose). In the controls, laser and applanation with BSS resulted in an average endothelial loss of 18% (n = 5, range of 14-26%). Applanation alone without laser dissection resulted in cell loss of 9% (n = 5, range of 7-12%) using BSS and 9% (n = 6, range 1-42%) Hydroxypropylmethyl-cellulose. CONCLUSIONS: The technique of using a viscoelastic "cushion" to protect endothelial cells from damage during posterior laser dissection prior to transplantation is promising. Viscoelastic coating protects the endothelial layer from damage from the coupling lens better than a layer of BSS. The lasering process, however, causes damage in addition to applanation with the laser lens. Further studies are warranted to optimize reproducibility of endothelial cell survival and evaluate the smoothness of stromal dissections in the posterior cornea.

A comparison of gatifloxacin to ciprofloxacin in the prophylaxis of Streptococcus pneumoniae in rabbits in a LASIK model.

PURPOSE: To investigate the efficacy of the fourth-generation fluoroquinolone, gatifloxacin 0.3%, compared to ciprofloxacin 0.3%, in preventing Streptococcus pneumoniae keratitis in a rabbit laser in situ keratomileusis (LASIK) model. METHODS: Twelve albino rabbits had bilateral lamellar flaps created. Group A (eight eyes) was given gatifloxacin 0.3%; group B (eight eyes) was given ciprofloxacin 0.3%; and group C (eight eyes) served as the controls. Groups A and B received one drop of antibiotic 20 minutes before the creation of the lamellar flap, at the conclusion of flap formation, and four times per day for 3 days. All corneas were inoculated with 0.1 mL of 4 x 10 organisms/mL of S. pneumoniae immediately after flap formation. On day 3, all corneas were examined and cultured. RESULTS: Group A (gatifloxacin) had no infiltrates and three areas of 1-mm central corneal haze. On day 3, one of eight corneas had a positive culture. Group B (ciprofloxacin) had seven infiltrates, including one perforation, and six of eight corneas had positive cultures. Group C (control) had eight corneal infiltrates, and all eight corneas had positive cultures. The data show a statistically significant difference between gatifloxacin and ciprofloxacin and gatifloxacin and control for mean infiltrate size and mean culture scores. CONCLUSIONS: The fourth-generation fluoroquinolone, topical gatifloxacin 0.3%, is superior to topical ciprofloxacin 0.3% for prophylaxis against a clinical isolate of S. pneumoniae in a rabbit LASIK model.

Penetration of topically applied gatifloxacin 0.3%, moxifloxacin 0.5%, and ciprofloxacin 0.3% into the aqueous humor.

PURPOSE: To investigate the aqueous penetration of 3 commercially available ophthalmic fluoroquinolones. DESIGN: Prospective, double-masked, clinical study. PARTICIPANTS: Fifty-two eyes of 52 patients. INTERVENTION: Fifty-two patients undergoing cataract surgery were given preoperative topical gatifloxacin 0.3% (Zymar), moxifloxacin 0.5% (Vigamox), or ciprofloxacin 0.3% (Ciloxan). The patients were instructed to use their antibiotic drops 4 times a day for 3 days before surgery. On the day of surgery, patients were given their assigned antibiotic every 15 minutes for 3 doses, 1 hour before their procedure. At the time of surgery, 0.1 ml of aqueous fluid was aspirated from the anterior chamber with an air cannula needle attached to a tuberculin syringe. The aspirate was immediately stored at -70 degrees C. MAIN OUTCOME MEASURE: Fluoroquinolone concentrations were determined by reverse-phase high-pressure liquid chromatography assay technique with ultraviolet detection at a wavelength of 275 nm. RESULTS: Mean aqueous concentration of gatifloxacin in 16 eyes was 0.63 microg/ml (standard deviation [SD], 0.30), moxifloxacin in 14 eyes was 1.31 microg/ml (SD, 0.46), and the mean concentration of ciprofloxacin in 22 eyes was 0.15 microg/ml (SD, 0.11). CONCLUSIONS: Both moxifloxacin (P<0.001) and gatifloxacin (P<0.005) penetrated the aqueous humor at significantly higher levels than ciprofloxacin. Moxifloxacin penetrated into the aqueous humor at significantly higher levels than gatifloxacin (P<0.05). The anterior chamber levels of moxifloxacin and gatifloxacin may be due to the difference in antibiotic concentration.

Comparative penetration of moxifloxacin and gatifloxacin in rabbit aqueous humor after topical dosing.

PURPOSE: To evaluate the aqueous penetration of the fourth-generation fluoroquinolones moxifloxacin and gatifloxacin. SETTING: University of Arizona, Tucson, Arizona, USA. METHODS: Forty eyes of 20 New Zealand white rabbits were divided into 2 experimental groups. In Experiment I rabbits (20 eyes), a commercial preparation of topical gatifloxacin 0.3% was administered to 9 eyes and moxifloxacin 0.5% to 9 eyes; 2 eyes served as a control. Eyes were dosed according to a keratitis protocol; ie, every 15 minutes for 4 hours. The aqueous humor was sampled 10 minutes after the last dose. Experiment II rabbits (20 eyes) were dosed according to a cataract prophylaxis protocol; ie, 4 times a day for 10 days. The aqueous humor was sampled 1 hour after the last dose of antibiotic in 12 eyes and 24 hours after the last dose in 8 eyes. High-performance liquid chromatography was used to determine the fluoroquinolone concentration. RESULTS: In the keratitis dosing protocol, the mean concentration of moxifloxacin in the aqueous (n=9) was 11.057 microg/mL (range 7.66 to 18.87 microg/mL), which was significantly higher than the mean concentration of gatifloxacin (n=8) (7.570 microg/mL [range 4.75 to 10.86 microg/mL]) (P=.030). In the cataract prophylaxis dosing protocol, the mean aqueous concentration of moxifloxacin (n=6) was 1.745 microg/mL (range 0.92 to 3.87 mg/mL). The mean concentration of gatifloxacin (n=6) was 1.207 microg/mL (range 0.44 to 2.44 microg/mL). The difference was not statistically significant (P=.359). CONCLUSIONS: Higher mean levels (x1.46) of aqueous penetration were achieved with moxifloxacin than with gatifloxacin in the keratitis-dosing model. There was no statistically significant difference between the 2 drugs in the cataract prophylaxis dosing model. Both antibiotics had aqueous levels in excess of the minimum inhibitory concentration for most pathogenic organisms in both models.

Aqueous penetration of gatifloxacin and levofloxacin into the rabbit aqueous humor following topical dosing.

The aqueous penetration of the commercial preparations of the fluoroquinolone antibiotics ofloxacin, ciprofloxacin, levofloxacin, and gatifloxacin were compared following topical dosing in a rabbit model. Levofloxacin achieved the highest aqueous concentrations, with a mean aqueous level of 4.8014 microcg/mL (p = 0.002, p = 0.00002, p = 0.015.) Ofloxacin (2.5136 microcg/mL) and gatifloxacin (2.4817 microcg/mL) achieved statistically equal aqueous concentrations (p = 0.479). Ciprofloxacin reached the lowest levels in the aqueous humor (0.9616 microcg/mL, p = 0.00002, 0.00004, 0.008). Gatifloxacin alone achieved concentrations in excess of the MIC90s of gram-positive pathogens of concern.

Wavefront and topography: keratome-induced corneal changes demonstrate that both are needed for custom ablation.

PURPOSE: To examine the effects of laser in situ keratomileusis (LASIK) flap incision and healing on the shape of the cornea and the wavefront error of the eye. METHODS: Four weeks prior to bilateral LASIK, study subjects had a flap cut using a keratome in one eye. The fellow eye remained untouched as a control. Corneal topography and wavefront errors were measured at 1 day, 1, and 4 weeks after the flap was created. After 4 weeks, the flap was lifted and LASIK was performed. The control eye also had LASIK at this time. RESULTS: Differences in corneal shape and wavefront error consistent with a mild hyperopic shift were seen as a result of the keratome incision. CONCLUSION: Cutting the flap in LASIK causes subtle changes to corneal shape and the optics of the eye that may affect customized treatments. Additional work is needed to quantify these changes so that their effect can be incorporated into future treatments.

Causes of blindness and visual impairment in a population-based sample of U.S. Hispanics.

OBJECTIVE: To describe the causes of blindness and visual impairment in a population-based sample of Hispanics. DESIGN: A cross-sectional study. PARTICIPANTS: A random sample of 4774 Hispanic residents of Santa Cruz and Pima Counties in Southern Arizona aged 40 years and older who participated in Proyecto VER (Vision Evaluation and Research). TESTING: Subjects were interviewed and underwent a thorough ophthalmic examination. Presenting and best-corrected visual acuity was determined using the Early Treatment of Diabetic Retinopathy Study protocol, followed by a standardized ophthalmic examination to determine the causes of visual loss. Anterior and posterior segment specialists in ophthalmology confirmed the causes. MAIN OUTCOME MEASURES: Causes of visual loss (best-corrected acuity worse than 20/40). RESULTS: The response rate of eligible participants was more than 70%. Best-corrected acuity in the better seeing eye worse than 20/40 increased from 0.3% in those aged 40 to 49 to 5.6% in those aged 65 and older. The leading cause was cataract, accounting for 42% of all visual loss, followed by age-related macular degeneration (15%), and diabetic retinopathy (13%). Among 14 people who were bilaterally blind, open-angle glaucoma was the leading cause. Women had higher age-adjusted prevalence of severe cataract compared with men and were more likely to be visually impaired from cataract, diabetic retinopathy, and open-angle glaucoma, although gender differences were not statistically significant. CONCLUSIONS: Causes of visual impairment differ from those reported in Caucasian populations, with open-angle glaucoma being the leading cause of blindness. Further work on gender-based obstacles to eye care in the Hispanic community may be warranted.