Untreated Control Eyes Research Articles

Anterior and posterior corneal stroma elasticity after corneal collagen crosslinking treatment

The purpose of this project was to assess anterior and posterior corneal stromal elasticity after corneal collagen cross linking (CXL) treatment in human cadaver eyes using Atomic Force Microscopy (AFM) through indentation. Twenty four human cadaver eyes (12 pairs) were included in this study and divided into 2 groups (6 pairs per group). In both groups, the left eye (OS) served as a control (no riboflavin or CXL treatment was performed) and the right eye (OD) underwent CXL treatment (30 min of riboflavin pretreatment followed by 30 min of exposure to 3 mW/cm2 of ultraviolet light). In group 1, the anterior stroma was exposed by manual delamination of approximately 50 μm of the corneal stroma including Bowman's membrane. In group 2, the posterior stroma was exposed by delamination of the anterior 50% of the corneal stroma including Bowman's membrane. Delamination was performed after crosslinking treatment in the case of the treated eyes. In all eyes, the stromal elasticity was quantified using AFM through indentation. Young's modulus of elasticity for the anterior cornea (group 1) was 245.9 ± 209.1 kPa (range: 82.3–530.8 kPa) for the untreated control eyes, and 467.8 ± 373.2 kPa (range: 157.4–1126 kPa) for the CXL treated eyes. Young's modulus for the posterior cornea (group 2) was 100.2 ± 61.9 kPa (range: 28.1–162.6 kPa) for the untreated control eyes and 66.0 ± 31.8 kPa (range: 31.3–101.7 kPa) for the CXL treated eyes. Young's modulus of the anterior stroma significantly increased after CXL treatment (p = 0.024), whereas the posterior stroma did not demonstrate a significant difference in Young's modulus after CXL treatment (p = 0.170). The anterior stroma was stiffer than the posterior stroma for both the control and CXL treatment groups (p = 0.077 and p = 0.023, respectively). Our findings demonstrate that stiffness of the anterior corneal stroma after CXL treatment seems to increase significantly, while the posterior stroma does not seem to be affected by CXL.

Progression of Lesion Size in Untreated Eyes With Exudative Age-Related Macular Degeneration

To test the hypothesis that the natural history of choroidal neovascularization lesion size is uniform across prior randomized controlled clinical trials of exudative age-related macular degeneration (AMD), with apparent differences arising from different entry times of eyes into clinical trials. We conducted a retrospective meta-analysis of control eye data from 5 age-related macular degeneration trials (Treatment of Age-Related Macular Degeneration with Photodynamic Therapy; Verteporfin in Photodynamic Therapy; VEGF Inhibition Study in Ocular Neovascularization; Minimally Classic/Occult Trial of the Anti-VEGF Antibody Ranibizumab in the Treatment of Neovascular Age-Related Macular Degeneration; and Phase 3b, Multicenter, Randomized, Double-masked, Sham Injection-controlled Study of the Efficacy and Safety of Ranibizumab in Subjects with Subfoveal Choroidal Neovascularization with or without Classic Choroidal Neovascularization Secondary to AMD Study), which were plotted on a double reciprocal plot of 1 / lesion size (disc area) vs 1 / time (months after enrollment). To account for the different entry times, we introduced a horizontal translation factor to shift each data subset until r2 was maximized for the cumulative trend line. Cumulative data for untreated control eyes fit a straight line on a double reciprocal plot (r2 = 0.98) after the introduction of horizontal translation factors. Our model predicts that a choroidal neovascular lesion will eventually enlarge to a size of 10.6 disc areas without treatment and that the lesion will reach half of its maximum size within 14.0 months after onset of exudation. The linear expansion rate of untreated lesions is approximately 26.0 μm per day for the smallest lesions and decreases gradually as the lesions enlarge. The pattern of choroidal neovascularization lesion size enlargement in AMD eyes is uniform across a wide range of clinical trials, with apparent differences arising from different entry times of patients into various trials. The main determinant of choroidal neovascularization lesion size enlargement is the duration of exudative disease.

Effect of Coherin™ on intraocular pressure, pupil size and heart rate in the glaucomatous Beagle: a pilot study

To evaluate effects of Coherin™ on intraocular pressure (IOP), pupil size (PS), and heart rate (HR) in glaucomatous Beagles in single-dose studies in a pilot study. Intraocular pressure, PS, and HR were measured in eight glaucomatous Beagles. One randomly chosen eye received single 50 μL doses of differing concentrations of Coherin™ (treated eye) or vehicle (placebo-treated eye), and the fellow eye served as the untreated control. After the first measurements, a single dose of either Coherin™ or sterile water vehicle was instilled in the drug and placebo eyes, respectively. The mean ± SEM diurnal changes in IOP after 0.005%, 0.01%, 0.2%, 0.284%, 1%, 2%, and 4% topical Coherin™ once daily were 7.6 ± 3.2 mmHg, 15.5 ± 5.3 mmHg, 11.2 ± 4.4 mmHg, 11.8 ± 4.4 mmHg, 19.1 ± 3.8 mmHg, 5.0 ± 1.8 mmHg, and 8.8 ± 2.8 mmHg, respectively. The declines in IOP were significantly different (P < 0.05) from the untreated control eyes with the 0.2% and 0.284% Coherin™-treated eyes and suggestive for 1% Coherin™ concentrations. No signs of irritation, significant PS, and HR changes were detected in the Coherin™-treated eyes. Of seven different concentrations, 2% and 0.248% Coherin™ produced significant declines in IOP in the glaucomatous beagle in single-dose studies when compared to both untreated control and placebo-treated eyes. One percent Coherin™ solution produced significant IOP decreases compared with the placebo-treated eye but not the untreated control eyes. No local ocular irritation, PS and HR changes were observed in Coherin™-treated eyes. This pilot study suggests that topical Coherin™ has potential as an ocular hypotensive agent.

Transepithelial corneal collagen crosslinking: Bilateral study

To evaluate the efficacy of transepithelial collagen crosslinking (CXL) in patients with bilateral progressive keratoconus. Outpatient ophthalmic clinic. Cohort study. Patients with a history of bilateral progressive keratoconus were recruited. The worst eye was treated with transepithelial CXL, while the fellow eye was left untreated as a control. Transepithelial CXL was performed by applying an enhanced riboflavin solution (riboflavin 0.1%, dextrane T500 with trometamol [Tris-hydroxymethyl aminomethane] and EDTA [ethylenediaminetetraacetic] sodium salt) on the intact corneal epithelium for 30 minutes before irradiation with ultraviolet A (370 nm at 3 mW/cm(2)) for 30 minutes. Follow-up was 18 months in all eyes. The study enrolled 20 patients. Transient hyperemia and mild foreign-body sensation occurred in 8 eyes (40%) after treatment; both resolved after 24 hours. In treated eyes, there were statistically significant improvements in uncorrected and corrected visual acuity and topography-derived keratometry, cone apex power, and higher-order aberrations (P<.05). In untreated control eyes, there was a general trend toward worsening of these parameters. No complications were reported. Transepithelial CXL treatment appeared to halt keratoconus progression, with a statistically significant improvement in visual and topographic parameters. The treatment was safe and well tolerated. Its noninvasive nature makes it potentially useful in cases in which epithelial debridement is ideally avoided, such as pediatric cases, uncooperative patients, and thin corneas with thicknesses nearing 380 μm.

Visual guidance of recovery from lens-induced myopia in tree shrews (Tupaia glis belangeri).

To examine, in tree shrews, the visual guidance of recovery from negative lens-induced myopia by measuring the effect of wearing low-power negative or positive lenses during recovery. To learn if removing a negative lens for 2 h per day, after compensation has occurred, is sufficient to produce recovery. Starting 16 days after natural eye opening (days of visual experience), juvenile tree shrews wore a monocular -5 D lens for 11 days to produce compensation (age-appropriate refraction while wearing the lens). Recovery in four groups was started by discontinuing -5 D lens wear, which caused the treated eyes to be refractively myopic, and substituting: no lens (n = 7), a plano lens (n = 8), a -2 D lens (n = 6) or a +2 D lens (n = 10). In a fifth group (n = 6), the -5 D lens was removed for 2 h each day but worn the remainder of the time. Non-cycloplegic refractive measurements were made daily for the first 10 days and then less frequently. After 31-35 days, the lens-guided recovery period was ended for most animals; periodic measures were continued to assess post-lens recovery changes. All the eyes responded to the -5 D lens and were myopic (-4.8 ± 0.1 D, mean ± S.E.M.) compared to the untreated fellow control eye. In all groups except the -2 D lens group, some animals exhibited slow or incomplete recovery. During recovery, the treated eye of most animals recovered until its refraction, measured with the recovery-lens in place, was near to that of the control eye. Measured without the lens, the -2 D group was myopic and the +2 D group was hyperopic. With the lens in place, the plano-lens, -2 D lens, and +2 D lens groups remained slightly myopic (-1.0 ± 0.3 D, -0.6 ± 0.2 D and -1.3 ± 0.1 D, respectively). The rate of recovery during the first four days was unrelated to the amount of myopia initially experienced by the recovering eyes. Removal of the -5 D lens for 2 h each day produced recovery. During recovery, the emmetropization mechanism uses the presence of myopia, but perhaps not the magnitude, to guide eyes toward a refractive state similar to the control eye, regardless of whether the optically-recovered eye is longer or shorter than the fellow control eye. Wearing a goggle frame containing a lens of any power limits the recovery. The recovery signal can be intermittent, present for only 2 h per day, and still mediate recovery in competition with increasing amounts of hyperopia as recovery progresses.

Ab-Externo AAV-Mediated Gene Delivery to the Suprachoroidal Space Using a 250 Micron Flexible Microcatheter

BackgroundThe current method of delivering gene replacement to the posterior segment of the eye involves a three-port pars plana vitrectomy followed by injection of the agent through a 37-gauge cannula, which is potentially wrought with retinal complications. In this paper we investigate the safety and efficacy of delivering adeno-associated viral (AAV) vector to the suprachoroidal space using an ab externo approach that utilizes an illuminated microcatheter.Methods6 New Zealand White rabbits and 2 Dutch Belted rabbits were used to evaluate the ab externo delivery method. sc-AAV5-smCBA-hGFP vector was delivered into the suprachoroidal space using an illuminated iTrackTM 250A microcatheter. Six weeks after surgery, the rabbits were sacrificed and their eyes evaluated for AAV transfection using immunofluorescent antibody staining of GFP.ResultsImmunostaining of sectioned and whole-mounted eyes demonstrated robust transfection in all treated eyes, with no fluorescence in untreated control eyes. Transfection occurred diffusely and involved both the choroid and the retina. No apparent adverse effects caused by either the viral vector or the procedure itself could be seen either clinically or histologically.ConclusionsThe ab externo method of delivery using a microcatheter was successful in safely and effectively delivering a gene therapy agent to the suprachoroidal space. This method presents a less invasive alternative to the current method of virally vectored gene delivery.

Corneal Wound Healing After Ultraviolet-A/Riboflavin Collagen Cross-Linking: A Rabbit Study

To investigate corneal wound healing following ultraviolet-A (UVA)/riboflavin corneal collagen cross-linking (CXL) in rabbit corneas. Thirty-six rabbits were enrolled in the study. Animals were divided into three treatment groups and corneas were analyzed at 24 hours and 4 weeks postoperatively. Thus, each group had 6 rabbits at each time point. Treatment groups were: 1) standard UVA+riboflavin CXL, 2) UVA alone, and 3) riboflavin alone. One eye of each rabbit served as an untreated control eye. TUNEL assay was performed to detect stromal cell apoptosis. Immunocytochemistry was performed to detect the inflammatory marker CD11b expressed in monocytes and the alpha-smooth muscle actin (SMA) marker expressed in myofibroblasts. At 24 hours, corneas from the UVA+riboflavin CXL group had significantly more apoptosis than the UVA alone and riboflavin alone groups. Eyes from all three groups had significantly more inflammatory cell influx into the cornea than unwounded controls. Four weeks after the procedure, many corneas in the UVA+riboflavin CXL group had mild haze, but very few SMA-positive myofibroblasts could be detected in the central cornea. Riboflavin+UVA CXL triggers more anterior keratocyte apoptosis than corneal scrape with UVA alone or riboflavin alone. Inflammation monitored by the monocyte marker CD11b was present, but not statistically different among the three groups. Very little myofibroblast generation could be detected after UVA+riboflavin CXL, indicating that the mild stromal haze associated with this procedure is normally related to transient corneal fibroblast generation rather than more persistent haze due to generation of myofibroblasts.

The effectiveness of an improved combination therapy for experimental Staphylococcus aureus keratitis

antibiotic and steroid combination therapies, such as tobramycin with dexamethasone, are often used in ophthalmology to treat or prevent infection and inflammation. The purpose of this study was to use a model of Staphylococcus aureus keratitis to quantify and compare the effectiveness of a standard tobramycin and dexamethasone combined therapy, with each drug individually, and with a new formulation of the two drugs in a xanthan gum vehicle. rabbit corneas were intrastromally injected with a methicillin-sensitive S. aureus (MSSA) or a methicillin-resistant S. aureus (MRSA) strain. Rabbit eyes were treated every hour from 10 to 15 hours postinfection (PI) with 0.1% dexamethasone, 0.3% tobramycin, 0.3% tobramycin with 0.1% dexamethasone, or 0.3% tobramycin with 0.05% dexamethasone in a xanthan gum vehicle (ST). Slit lamp examinations (SLE) were performed on infected eyes and pathology scored at 15 hours PI. At 16 hours PI, colony forming units (CFUs) per cornea were quantified. the CFUs in eyes treated with dexamethasone alone were similar to untreated control eyes for MSSA or MRSA infections. All other treatment groups had significantly less CFUs per cornea than untreated eyes. The eyes treated with the ST formulation had significantly fewer CFUs per cornea than all other treatment groups when infected with MSSA or MRSA. The SLE scores of MSSA or MRSA infected eyes treated with tobramycin alone were similar to untreated control eyes. All other treatment groups had significantly lower SLE scores than untreated controls eyes, but were not significantly different from each other. the results of this study demonstrated that the tobramycin and dexamethasone combination therapy with a xanthan gum vehicle has an improved bactericidal effectiveness compared to the commercially available formulation, and maintains a similar anti-inflammatory effect while containing half the amount of steroid.

Eyedrops Containing SA9000 Prodrugs Result in Sustained Reductions in Intraocular Pressure in Rabbits

Poor topical bioavailability and ocular irritation have impeded the development of the diuretic, ethacrynic acid (ECA) as a clinically useful ocular hypotensive for the treatment of glaucoma. Thus, the development of analogs and prodrugs of analogs with improved ocular penetration, potency, and tolerability is required. The aim of this work is to evaluate the corneal penetration and ocular distribution of SA9000, an ECA analog. Novel SA9000 prodrugs intended to further improve ocular pharmacodynamic effect were also evaluated. SA9000 penetrated porcine corneas more effectively than ECA in corneal diffusion studies. In vivo studies in Dutch-belted (DB) rabbits indicated that topical application of a single dose (0.3%) of SA9000 could significantly reduce intraocular pressure (IOP) (approximately 25% vs. fellow untreated eye) but caused significant conjunctival hyperemia. Since this hyperemia was likely the result of its inherent thiol reactivity, SA9000 was formulated with equimolar cysteine, an exogenous thiol donor. The administration of increasing SA9000-cysteine adduct concentrations (0.3%, 0.6%, 0.9%) demonstrated that they cause less ocular irritation than unadducted SA9000 but could still significantly reduce IOP (0.3%: 8.7 +/- 2%; 0.6%: 14.4 +/- 5%; 0.9%: 23.3 +/- 4.4%) versus untreated contralateral control eyes. These data suggest that novel thiol donor adduction can improve the ocular bioavailability and tolerability of SA9000. SA9000-cysteine prodrugs may represent a new option for the topical treatment of glaucoma.

Subfoveal exudative age-related macular degeneration: evidence for preoccult disease

A careful analysis of randomized clinical trials on exudative age-related macular degeneration reveals apparent differences in behavior of untreated control eyes among these trials. A priori there are two possible explanations: each study contains a unique subpopulation of patients; or apparent differences arise from differences in time of entry of these eyes into clinical trials. To correctly account for differences in time of entry into clinical trials, we introduced a horizontal translation factor, expressed in months, to shift each data subset horizontally to maximize r2 for the cumulative trend line; this increases the overall r2 to 0.95, demonstrating that most of the variation in visual acuity over time is explained by the initial visual acuity. This analysis also suggests that occult disease is an earlier stage than minimally classic and predominantly classic disease and that there is a subclinical stage of subfoveal exudation ('preoccult') for patients with age-related macular degeneration. The pattern of vision loss experienced in age-related macular degeneration eyes with subfoveal choroidal neovascularization is uniform across a wide range of clinical trials, with apparent differences in initial and final visual acuity in untreated eyes arising from differences in the time of entry into clinical trials. Our data suggest that visual loss may occur during a preoccult phase of choroidal neovascularization, prior to the development of occult disease.

Correction of the disease phenotype in the mouse model of Stargardt disease by lentiviral gene therapy

Autosomal recessive Stargardt disease (STGD1) is a macular dystrophy caused by mutations in the ABCA4 (ABCR) gene. The disease phenotype that is most recognized in STGD1 patients, and also in the Abca4-/- mouse (a disease model), is lipofuscin accumulation in retinal pigment epithelium. Here, we tested whether delivery of the normal (wt) human ABCA4 gene to the subretinal space of the Abca4 -/- mice via lentiviral vectors would correct the disease phenotype; that is, reduce accumulation of the lipofuscin pigment A2E. Equine infectious anemia virus (EIAV)-derived lentiviral vectors were constructed expressing either the human ABCA4 gene or the LacZ reporter gene under the control of the constitutive (CMV) or photoreceptor-specific (Rho) promoters. Abca4-/- mice were injected subretinally with 1 microl ( approximately 5.0 x 10(5) TU) of each EIAV vector in one eye at postnatal days 4 and 5. An injection of saline, an EIAV-null vector, or an uninjected contralateral eye served as a control. Mice were killed at various times after injection to determine photoreceptor (PR) transduction efficiency and A2E concentrations. EIAV-LacZ vectors transduced from 5 to 20% of the PRs in the injected area in mice. Most importantly, a single subretinal injection of EIAV-CMV-ABCA4 to Abca4-/- mouse eyes substantially reduced disease-associated A2E accumulation compared to untreated and mock-treated control eyes. Treated eyes of Abca4-/- mice accumulated 8-12 pmol per eye (s.d.=2.7) of A2E 1 year after treatment, amounts comparable to wt controls, whereas mock-treated or untreated eyes had 3-5 times more A2E (27-39 pmol per eye, s.d.=1.5; P=0.001-0.005). Although extrapolation to humans requires caution, the high transduction efficiency of both rod and cone photoreceptors and the statistically significant reduction of A2E accumulation in the mouse model of STGD1 suggest that lentiviral gene therapy is a potentially efficient tool for treating ABCA4-associated diseases.

Introduction of silencing-inducing transgene against Fgf19 does not affect expression of Tbx5 and β3-tubulin in the developing chicken retina

Fgf19 is known to be expressed in the developing chicken eye but its functions during retinal development have remained elusive. Since Fgf19 is expressed in the dorsal portion of the optic cup, it is intriguing to know whether FGF19 is required for expression of dorso-ventral morphogenetic genes in the eye. To clarify this, expression patterns of Tbx5 and Vax were examined in the developing eye after in ovo RNA interference targeted against Fgf19. Quantitative polymerase chain reaction (PCR) analysis showed that the short-hairpin RNAs (shRNAs) targeted against Fgf19 could reduce its expression in the eye to less than 50% of a relative amount of mRNA, compared with contralateral or untreated control eyes. However, no obvious alteration in expression domains of Tbx5 or Vax was observed. Misexpression of Tbx5 or Tbx5-RNAi did not alter the Fgf19 expression either. Furthermore, although Fgf19 is expressed in the central retina before neurogenesis occurs, beta3-tubulin, a marker for early retinal differentiation was still detected in the central retina after knockdown of Fgf19. Thus, knockdown of Fgf19 supports no obvious regulations between Fgf19 and Tbx5, or exhibits no phenotypes that perturb early retinal differentiation.

Progressive Visual Loss in Subfoveal Exudation in Age-related Macular Degeneration: A Meta-analysis Using Lineweaver-Burke Plots

To analyze the randomized clinical trials in exudative age-related macular degeneration (AMD) to reveal apparent differences in the behavior of untreated control eyes among these trials. Herein we test the hypothesis that the behavior of untreated control eyes is actually the same in all studies, with apparent differences arising from differences in the time of entry of eyes into clinical trials. Retrospective meta-analysis of prior clinical trials. Control eye data from six AMD studies (Macular Photocoagulation Study, Subfoveal Surgery Trial, Photodynamic Therapy [TAP] With Visudyne, pegaptanib trial for neovascular AMD, anecortave acetate trial, and 360 degree Macular Translocation Study) were plotted on a double reciprocal plot of 1/(Letters Lost) vs 1/(Months After Enrollment). To account for differences in time of entry into clinical trials, we introduced a horizontal translation factor to shift each data subset horizontally to maximize r(2) for the cumulative trend line. Cumulative data for untreated control eyes fits a straight line on a double reciprocal plot (r(2) = .9521); an untreated eye would eventually deteriorate to a final vision of 20/640. The slope of the line predicts that patients would experience half of the maximum final vision within 10.88 months after exudation onset. The pattern of vision loss experienced in AMD eyes with subfoveal neovascularization is uniform across a wide range of clinical trials, with apparent differences arising from differences in the time of entry of patients into clinical trials.

SAFETY OF INTRAVITREAL INJECTION OF BEVACIZUMAB IN RABBIT EYES

To evaluate the safety of intravitreal injection of bevacizumab in rabbits using electrophysiological testing and histopathologic analysis. New Zealand albino rabbits were injected in one eye with control antibody (n = 2), 0.05 mL of bevacizumab (n = 3), or 0.2 mL of bevacizumab (n = 3). Electroretinograms were obtained 1 week and 4 weeks after injection. Histologic analysis was performed after completion of the electroretinographic studies. No statistical differences were seen in scotopic and photopic a- and b-wave amplitudes between untreated control and bevacizumab-injected eyes. No histopathologic differences were identified between untreated control and bevacizumab-injected eyes. Our study did not find evidence of retinal toxicity from a single intravitreal injection of bevacizumab in rabbits.

Protective Effect of Hepatocyte Growth Factor Against Degeneration of the Retinal Pigment Epithelium and Photoreceptor in Sodium Iodate–Injected Rats

Purpose: To investigate the possible protective effect of hepatocyte growth factor (HGF) against degeneration of photoreceptors and retinal pigment epithelium (RPE) in vivo. Methods: Sprague-Dawley (SD) rats received an intravitreal injection of HGF in the right eye. The left eye was injected with vehicle as a control. Two days after the intravitreal injections, rats were administered 40 mg/kg of sodium iodate (NaIO3) intravenously. Scotopic ERGs were elicited by different stimulus intensities with a maximum luminance of 0.84 log cds/m2. To evaluate RPE function, the azide response was evoked by intravenous injection of 0.1 mg sodium azide. These electrophysiological measurements were conducted on days 4, 7, 14, and 28 after the NaIO3 injections. After recording ERGs or azide response, animals were sacrificed for quantification of the histological change and immunohistochemical analysis using antibodies against RPE 65. Results: The threshold for the scotopic b-wave was significantly lower in HGF-treated eyes than in untreated control eyes (p < 0.005), and maximum b-wave amplitudes (Vbmax) were significantly larger in HGF-treated eyes (p < 0.05) across all experimental time points after NaIO3 injection. Azide response amplitudes were significantly larger in the HGF-treated eyes than in the untreated eyes (p < 0.05). The structure of the outer retina was preserved to a greater degree in the HGF-treated eyes than in the untreated eyes (p < 0.05). Immunohistochemical analysis demonstrated that irregular alignment of the outer nuclear layer was confined to the retinal area that was not stained with RPE 65. Conclusions: Our results indicated that an intravitreal injection of HGF provided significant protection against degeneration of the photoreceptor and RPE induced by systemic administration of NaIO3. This suggests that HGF could be used as a therapeutic agent for degeneration of photoreceptors as well as RPE.

Changes in bacterial and fungal ocular flora of clinically normal horses following experimental application of topical antimicrobial or antimicrobial-corticosteroid ophthalmic preparations

To determine effects of topical antimicrobial and antimicrobial-corticosteroid preparations on the ocular flora of horses. animals: 40 horses. One eye was treated 3 times daily for 2 weeks with one of the following ointments: (1) neomycin-bacitracin-polymyxin B, (2) 0.6% prednisolone-0.3% gentamicin, (3) neomycin-polymyxin B-0.05% dexamethasone, or (4) treated (artificial tears) control. Contralateral eyes of treated control eyes served as untreated control eyes. Corneal and conjunctival specimens for bacterial and fungal cultures were collected prior to initiation of treatment, after 1 and 2 weeks of treatment, and 2 weeks after concluding treatment. Changes in culture growth quantity scores of bacterial and fungal species were analyzed. The most common species before treatment were the following: gram-positive bacteria included Streptomyces spp (66%), Staphylococcus spp (46%), Bacillus spp (32%), and Streptococcus spp (32%); gram-negative bacteria included Moraxella spp (28%), Escherichia coli (24%), Acinetobacter spp (18%), and Enterobacter spp (14%); and fungi included Aspergillus nidulans (56%), Cladosporium spp (32%), and Aspergillus fumigatus (22%). In all groups, the percentage of positive bacterial culture results, growth quantity score of gram-positive bacteria, and number of bacterial species isolated decreased at week 1 and increased at week 2, whereas growth quantity score of gram-negative bacteria decreased throughout treatment. Differences were not significant among groups. Fungal growth quantity score decreased during treatment in all groups. Repopulation of bacterial and fungal species occurred. All interventions decreased the number of microorganisms. Repopulation of normal flora occurred during and after treatment.

Analysis of irises with a latanoprost-induced change in iris color

To study the histologic aspects of irises subjected to extended latanoprost treatment. Prospective, observer-masked study. Iris biopsies of eyes treated with latanoprost were analyzed (all had a photographically documented increase in iris pigmentation) plus control eyes (untreated with prostanoids) using optical microscopy. PATIENT OR STUDY POPULATION: There were 14 study eyes treated with latanoprost and 8 untreated control eyes. The morphologic characteristics of the irises. The irises treated with latanoprost had an increased number of melanocytes with nuclear inclusions, granules of melanin in the vascular walls and the melanocytes and free granules in the stroma compared with control eyes (P = .001, P = .01, P = .004, P = .01, respectively, by the chi(2) test). Chronic therapy with latanoprost appears to induce more changes in the iris than a simple increase in the melanin content of the melanocytes.

Chronic ischemia induces regional axonal damage in experimental primate optic neuropathy.

To evaluate the effects of chronic optic nerve ischemia in a nonhuman primate model and to evaluate the regional variability of axonal loss. Unilateral ischemic optic neuropathy was induced by administration of endothelin-1 to the retrobulbar space via osmotic pumps in 12 primates for 6 to 12 months. The transversely cut sections were stained and divided into 16 regions. Average axonal density in each region was quantified and compared with the untreated contralateral control eyes. Mean axonal density was 208 310/mm(2) and 220 661/mm(2) in treated and control eyes, respectively (P = .03, 1-tailed paired t test), for the entire group. Two-way analysis of variance showed a significant effect of endothelin-1 on overall axonal density for the experimental group (P<.001). Among the nerves with significant axonal loss, the mean axonal loss was 11.6% (4%-21%). Regional mapping of the damage showed the axonal loss varied in the damaged nerves; the damaged regions often clustered within specific quadrants. Chronic ischemia induced by local administration of endothelin-1 causes significant loss of optic nerve axons with varying regional susceptibility. Clinical Relevance Localized damage occurs in other types of optic neuropathy, such as glaucoma, and may result from regional differences in anatomy, metabolism, or vasculature of the primate optic nerve.

Histological effect and protein expression in subthreshold transpupillary thermotherapy in rabbit eyes.

To investigate the histological effect of subthreshold transpupillary thermotherapy (TTT) on the retina. We performed TTT in normal pigmented rabbit eyes using an 810-nm diode laser with spot size of 1.2 mm, power of 50 mW, and varying durations of 15, 30, or 60 seconds. Four weeks later, fluorescein angiography was performed, and the enucleated eyes were examined by means of electron microscopy and immunohistochemical staining. Funduscopy immediately and at 4 weeks showed no discernable changes at TTT sites, and fluorescein angiography at 4 weeks showed no abnormalities. However, electron microscopy showed photoreceptor and retinal pigment epithelium cell disruption, changes more prominent with longer durations of treatment. Immunohistochemical staining was positive for heat shock protein 60, heat shock protein 70, tumor necrosis factor alpha, and vascular cell adhesion molecule 1 in the photoreceptors and retinal pigment epithelium at TTT sites. Untreated control eyes showed no staining. Despite the absence of changes evident by funduscopy and fluorescein angiography, TTT resulted in dose-dependent histological changes in photoreceptors and retinal pigment epithelium. The induction of heat shock proteins, cytokines, and cell adhesion molecules may play a role in the tissue response to subthreshold TTT. Clinical Relevance Unrecognized damage to the retina and retinal pigment epithelium may contribute to visual loss in eyes that undergo subthreshold TTT.

Time-specific intraocular pressure curves in Rhesus macaques (Macaca mulatta) with laser-induced ocular hypertension.

To detect and categorize time-specific variations in daytime intraocular pressure (IOP) found in Rhesus monkeys with laser-induced ocular hypertension. Ten male monkeys with argon laser-induced ocular hypertension in one eye were anesthetized with ketamine hydrochloride, and the IOP measured in both eyes at 7 a.m., 7.30 a.m., and then hourly until 1 p.m. with a Tonopen trade mark XL applanation tonometer. Intraocular pressure time profiles for both eyes in each animal were developed. The means +/- SD of the IOPs for both eyes were calculated for the whole 6-h study period, and the values compared statistically. The difference between the lasered eye mean IOP standard deviation and the normal eye mean IOP standard deviation for each animal during the 6-h follow-up was also calculated and compared. Mean IOP (+/- SD) in the glaucoma and normal eyes for the 10 animals during the 6-h study was 32.6 +/- 2.5 and 14.9 +/- 2.5 mmHg, respectively. The IOP was significantly higher in the experimental eye than in the normal eye (P = 0.0008). The mean IOP in the lasered eye did not significantly change during the study period, whereas a slight but significant increase in IOP of the normal eye over the study period was recorded (P = 0.003). The variance in IOP in the hypertensive eyes was considerably greater than that in the untreated control eyes. From 7 a.m. to 1 p.m. the IOP declined in five eyes and increased in the other five eyes with laser-induced ocular hypertension. The time-specific IOP variation pattern in the daytime in the laser treated eyes is significantly greater than the variation in the normotensive eyes. This shows that in order to detect statistical differences between IOP variations induced by an IOP-reducing drug, and the exaggerated spontaneous IOP variations present in the laser-induced hypertensive eye, sufficient animals should be included in any study. Understanding the time-specific IOP variation present in a group of monkeys with laser-induced ocular hypertension is essential prior to using the model for the evaluation of IOP-reducing drugs.