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Objectives: To test the hypotheses that lipopolysaccharide (LPS) stimulates leukotriene B4 (LTB4) production in human ocular surface and adenexal cells, arachidonic acid duplicates the stimulatory effect of LPS, LPS-binding protein potentiates LPS-induced LTB4 secretion, and dihydrotestosterone attenuates the immune effect of LPS., Methods: Immortalized human corneal, conjunctival, and meibomian gland epithelial cells were cultured in the presence or absence of fetal bovine serum and were exposed to vehicle, LPS, LPS plus LPS-binding protein, arachidonic acid, or dihydrotestosterone. Culture media were processed for the LTB4 analysis., Results: Lipopolysaccharide stimulates time-dependent secretion of LTB4 by human corneal, conjunctival, and meibomian gland epithelial cells. This effect, which we could not detect with arachidonic acid, is potentiated by exposure to LPS-binding protein. This potentiation, in turn, is significantly reduced by cellular treatment with dihydrotestosterone., Conclusions: Ocular epithelial cells have the ability to generate LTB4 in response to LPS exposure. This proinflammatory process is modulated by LPS-binding protein and by dihydrotestosterone., Clinical Relevance: When induced by appropriate stimuli, LTB4 production may have a role in the generation of inflammation in ocular surface disease.

Objective: Vernal keratoconjunctivitis (VKC) is a chronic allergic inflammatory disease with unclear etiology and pathogenesis. We investigated the tear film proteome of patients with VKC to understand the pathologic characteristics of VKC., Methods: Tear samples were collected from healthy volunteers and patients with VKC. Electrophoresis was performed to display the tear proteomic profiles according to VKC severity. The identities of differentially expressed proteins were analyzed by mass spectrometry and quantified by enzyme-linked immunosorbent assay. Impression cytology was performed on VKC conjunctival samples to demonstrate the cellular protein expression. Allergic sensitization was performed in mice to study the pathologic role of these proteins in VKC., Results: Hemopexin, an inflammatory protein, was elevated in the tear film of patients with VKC. The increased hemopexin concentration in VKC tears was significantly associated with disease severity. Impression cytology showed specific high hemopexin expression in dekeratinized conjunctival epithelium and necrotic macrophages in patients with VKC. Immunohistochemical examination of normal lacrimal tissues from mice showed that hemopexin was not expressed in any lacrimal apparatus. Under systemic and topical sensitization and challenge using hemopexin in mice, the affected eye had mild to moderate bead discharge, chemosis, and edema with excessive macrophage infiltration and conjunctival necrosis., Conclusion: An association exists between tear hemopexin and the development and pathologic effects of VKC., Clinical Relevance: Increased hemopexin may have a role in the development of VKC.

Objective: To evaluate the effect of a thiolated polymer lubricant, chitosan-N-acetylcysteine conjugate (C-NAC), in a mouse model of dry eye., Methods: Eye drops containing 0.5% C-NAC, 0.3% C-NAC, a vehicle (control group), artificial tears, or fluorometholone were applied in a masked fashion in a mouse model of induced dry eye from 3 days to 4 weeks after botulinum toxin B injection. Corneal fluorescein staining was periodically recorded. Real-time reverse transcriptase-polymerase chain reaction and immunofluorescence staining were performed at the end of the study to evaluate inflammatory cytokine expressions., Results: Mice treated with C-NAC, 0.5%, and fluorometholone showed a downward trend that was not statistically significant in corneal staining compared with the other groups. Chitosan-NAC formulations, fluorometholone, and artificial tears significantly decreased IL-1beta (interleukin 1beta), IL-10, IL-12alpha, and tumor necrosis factor alpha expression in ocular surface tissues., Conclusions: The botulinum toxin B-induced dry eye mouse model is potentially useful in evaluating new dry eye treatment. Evaluation of important molecular biomarkers suggests that C-NAC may impart some protective ocular surface properties. However, clinical data did not indicate statistically significant improvement of tear production and corneal staining in any of the groups tested., Clinical Relevance: Topically applied C-NAC might protect the ocular surface in dry eye syndrome, as evidenced by decreased inflammatory cytokine expression.

Objective: To create and test a slow-release antifibrotic drug-coated glaucoma drainage device using in vitro and in vivo experiments., Methods: A slow-release device incorporating mitomycin C in poly(2-hydroxyethyl methacrylate) disks was developed using redox-polymerization techniques. A standardized preparation of this drug delivery device was attached to the Ahmed glaucoma valve (model FP7; New World Medical, Inc, Rancho Cucamonga, California). Semicircular disks (5 x 6 mm) of P(HEMA)-mitomycin C containing varying concentrations of mitomycin C per gram dry weight of the gel were attached to the lower half of an Ahmed glaucoma valve plate. Water was pumped through the modified Ahmed glaucoma valve at a rate comparable to that of aqueous humor outflow, and mitomycin C release was measured. Modified and unmodified Ahmed glaucoma valves were implanted in a rabbit model, and drug release and fibrosis were assessed after 3 months., Results: The P(HEMA)-mitomycin C device released mitomycin C in vitro over 1 to 2 weeks. Studies in rabbits revealed that mitomycin C was released from the disks during the 3-month implantation. Histologic analysis demonstrated a significant reduction in inflammatory reaction and fibrosis in the resulting blebs., Conclusion: Our slow-release drug-coated glaucoma drainage device decreased fibrosis and inflammation in the resulting bleb in a rabbit model., Clinical Relevance: This device could reduce the failure rate of glaucoma drainage devices.

Objective: To investigate the inhibitory effect of solid hyaluronic acid-carboxymethyl cellulose film (hyaluronic acid film) on the formation of postoperative wound adhesion on rabbit eyes., Methods: We first created a conjunctival flap under which hyaluronic acid film was inserted. Then, we performed trabeculectomy on other rabbit eyes with hyaluronic acid film applied under and above the scleral flaps. Expression of proliferative cell nuclear antigen and alpha-smooth muscle actin (alpha-SMA) were histologically and immunohistochemically examined., Results: Hyaluronic acid film significantly prevented adhesions after both kinds of surgery. Particularly, subconjunctival scar formation was significantly inhibited when the film was simply inserted under the wound. Furthermore, the adhesion around the scleral flap of trabeculectomy was less formed in eyes treated with hyaluronic acid film than in control eyes. Immunoreactivity to proliferative cell nuclear antigen almost disappeared after 28 days postoperatively in both treated and control groups. The alpha-SMA-positive cells appeared much less around the film-treated wound than the control eye., Conclusion: The present results indicate that hyaluronic acid film can inhibit the formation of postoperative adhesion around the conjunctiva and sclera., Clinical Relevance: The results of this study indicate that this substance has potential benefits for improving ophthalmic surgery, such as filtering surgery for glaucoma.

Objective: To study the effect of topical application of very late antigen 4 (VLA-4) small-molecule antagonist (anti-VLA-4 sm) in a mouse model of dry eye disease., Methods: Anti-VLA-4 sm (or control vehicle) was applied topically to mice placed in a controlled-environment chamber. Corneal fluorescein staining and conjunctival T-cell enumeration were performed in the different treatment groups. Real-time polymerase chain reaction was used to quantify expression of inflammatory cytokines in the cornea and conjunctiva., Results: Dry eye syndrome induced increased corneal fluorescein staining, corneal and conjunctival tumor necrosis factor alpha messenger RNA expression, and T-cell infiltration into the conjunctiva. Very late antigen 4 blockade significantly decreased corneal fluorescein staining compared with the untreated dry eye disease and control vehicle-treated groups (P < .001 and P = .02, respectively). In addition, VLA-4 blockade was associated with a significant decrease in conjunctival T-cell numbers (P < .001 vs control vehicle-treated group) and tumor necrosis factor-alpha transcript levels in the cornea (P = .04 vs control vehicle-treated group) and conjunctiva (P = .048 vs control vehicle-treated group)., Conclusion: Application of topical anti-VLA-4 sm led to a significant decrease in dry eye signs and suppression of inflammatory changes at the cellular and molecular levels., Clinical Relevance: Topical blockade of VLA-4 may be a novel therapeutic approach to treat the clinical signs and inflammatory changes accompanying dry eye disease.

Objective: To study the effects of artificial tear viscosity on tear film thickness, upper and lower tear menisci, and tear volume using optical coherence tomography., Methods: The central tear film and tear menisci before and immediately after the instillation of different artificial tears were imaged in 40 eyes of 20 healthy individuals. Carboxymethylcellulose sodium, 1.0% (viscosity, 70 cP), propylene glycol, 0.3%, and polyethylene glycol, 0.4% (10 cP), carboxymethylcellulose, 0.5% (3 cP), and isotonic sodium chloride solution (1 cP) were tested on 2 consecutive days. All measurements, including tear film thickness, the height, radius, and area of the tear meniscus, and the estimated tear volume, were obtained at 0, 5, 20, 40, and 60 minutes after instillation., Results: At instillation, all artificial tears and isotonic sodium chloride solution caused an increase in all tear variables (P < .001). Tear film thickness remained significantly elevated for all drops at 5 minutes (P < .001) and returned to baseline at 20 minutes. Other variables returned to baseline at 5 minutes. Comparing the different drops, tear film thickness and lower meniscus variables at instillation were increased with the more viscous drops (P < .05)., Conclusion: Optical coherence tomography demonstrated that all tear preparations, including isotonic sodium chloride solution, increased tear film thickness for at least 5 minutes and other variables immediately after instillation.

Objective: To study the efficacy of topical application of alpha-linolenic acid (ALA) and linoleic acid (LA) for dry eye treatment., Methods: Formulations containing ALA, LA, combined ALA and LA, or vehicle alone, were applied to dry eyes induced in mice. Corneal fluorescein staining and the number and maturation of corneal CD11b(+) cells were determined by a masked observer in the different treatment groups. Real-time polymerase chain reaction was used to quantify expression of inflammatory cytokines in the cornea and conjunctiva., Results: Dry eye induction significantly increased corneal fluorescein staining; CD11b(+) cell number and major histocompatibility complex Class II expression; corneal IL-1alpha and tumor necrosis factor alpha (TNF-alpha) expression; and conjunctival IL-1alpha, TNF-alpha, interferon gamma, IL-2, IL-6, and IL-10 expression. Treatment with ALA significantly decreased corneal fluorescein staining compared with both vehicle and untreated controls. Additionally, ALA treatment was associated with a significant decrease in CD11b(+) cell number, expression of corneal IL-1alpha and TNF-alpha, and conjunctival TNF-alpha., Conclusions: Topical ALA treatment led to a significant decrease in dry eye signs and inflammatory changes at both cellular and molecular levels., Clinical Relevance: Topical application of ALA omega-3 fatty acid may be a novel therapy to treat the clinical signs and inflammatory changes accompanying dry eye syndrome.

Objective: To characterize chemokine receptor CCR5 expression on the conjunctival epithelium in dry eye syndromes., Methods: Conjunctival impression cytology samples were obtained from normal subjects (n = 15) and patients with dry eye syndrome (n = 45). Cells were harvested from impression cytology samples, and flow cytometry was performed to quantitatively analyze the cell surface expression of chemokine receptor CCR5. Characterization of CCR5-positive cells was done by 2-color flow cytometry using fluorescein-conjugated anti-CCR5 and phycoerythrin-conjugated anti-CD45 antibodies (where CD45 is a marker for bone marrow-derived cells). To study CCR5 messenger RNA transcripts, real-time polymerase chain reaction was done on RNA isolated from the impression cytology samples of normal subjects (n = 5) and patients with dry eye syndrome (n = 14)., Results: We found significant up-regulation in cell surface expression of CCR5 in patients with both aqueous tear-deficient and evaporative forms of dry eye syndrome (P<.001). The real-time polymerase chain reaction results (for messenger RNA) corroborated the flow cytometry data (for protein). The majority of the cells expressing CCR5 were non-bone marrow-derived resident epithelial cells of the conjunctiva., Conclusion: Our findings suggest that CCR5 up-regulation is significantly associated with dry eye syndrome-associated ocular surface disease. Clinical Relevance Chemokine receptor CCR5 or its ligands may serve as useful targets for modulation of tissue immunoinflammatory responses in dry eye syndromes.

Background: There is growing evidence that autonomic innervation is involved in the pathogenesis of mucus hypersecretion, goblet cell hyperplasia, and conjunctival hyperreactivity., Objective: To determine the expression of neurotransmitters and neurotransmitter receptors in vernal keratoconjunctivitis (VKC) tissues to evaluate whether neurogenic inflammation plays a role in this ocular atopic-related disorder., Methods: Biopsy specimens of upper tarsal conjunctiva from 8 VKC patients with active inflammation and from 4 healthy subjects were processed for immunohistochemistry using anti-M1, anti-M2, and anti-M3 muscarinic receptors; beta1-adrenergic receptor; vasoactive intestinal peptide; nerve growth factor; and protein gene product 9.5, a marker of nerve fibers., Results: In the conjunctival epithelium of VKC patients, M1 muscarinic receptor, nerve growth factor, and protein gene product 9.5 expression were decreased, whereas M2 and M3 muscarinic receptors and beta1-adrenergic receptor were irregularly distributed, compared with control subjects. Neurotransmitter receptors and vasoactive intestinal peptide expression were increased in the substantia propria-localized infiltrate of VKC compared with healthy tissue. Nerve growth factor and protein gene product 9.5 staining was also enhanced in the conjunctival stroma of VKC vs healthy conjunctiva., Conclusions: The inflamed conjunctiva of VKC patients demonstrated an obvious alteration in muscarinic and beta1-adrenergic receptor, vasoactive intestinal peptide, protein gene product 9.5, and nerve growth factor expression. These results substantiate the involvement of an autonomic dysfunction in the pathogenesis of VKC.

Objective: To evaluate status of conjunctival fibrillin-1 in patients with Marfan syndrome with ectopia lentis., Methods: Frozen sections of conjunctiva from 6 patients with Marfan syndrome with ectopia lentis and from 15 age-matched control subjects were stained with mouse antihuman fibrillin-1 antibody, using an avidin biotin immunoperoxidase technique. The fibrillin-1 staining characteristics of conjunctiva were analyzed with the light microscope., Results: All the fresh frozen sections of conjunctival samples from control subjects demonstrated a characteristic pattern of fibrillin-1 staining. We observed a woven network of thin fibrils of uniform thickness surrounding collagen bundles. The fresh frozen samples from patients with Marfan syndrome showed consistent qualitative differences in fibrillin-1 staining when compared with samples from control subjects. The fibrils were longer and straighter than normal, varied in caliber, and showed fewer tendencies to form a woven pattern., Conclusions: Consistent, qualitative abnormalities in fibrillin-1 staining pattern can be seen in the conjunctiva of patients with Marfan syndrome with ectopia lentis. Conjunctival biopsy deserves further investigation as a diagnostic modality for Marfan syndrome in patients with ectopia lentis.

Objective: To examine whether women with premature ovarian failure (POF) have abnormal findings in ocular surface or tear parameters and whether they report symptoms of ocular discomfort compared with age-matched controls., Methods: Sixty-five patients with POF and 36 age-matched healthy controls were examined for signs and symptoms of dry eye. The Ocular Surface Disease Index questionnaire and the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ 25) were administered to the participants. Assessments of ocular surface damage (Oxford and van Bijsterveld scores of vital dye staining) and tear status (Schirmer tests 1 [without anesthesia] and 2 [with anesthesia] and tear breakup time) were performed., Results: Women with POF scored significantly worse than controls on all ocular surface damage parameters: Oxford score (3.2 vs 1.7; P =.001), conjunctival lissamine green (2.1 vs 1.3; P =.02), corneal fluorescein staining (1.2 vs 0.4; P =.005), and van Bijsterveld score (2.1 vs 1.3; P =.02). Further, the proportion of patients with POF meeting the dry eye diagnostic criterion of a van Bijsterveld score greater than or equal to 4 was significantly greater among women with POF than among controls (20% vs 3%; P =.02). The POF group also tended to have worse scores than controls on self-reported symptoms, as measured by the overall Ocular Surface Disease Index (12.5 vs 2.1; P<.001) and the overall NEI-VFQ (94 vs 98; P =.001) after adjustment for age and race. Schirmer test scores and tear breakup time did not differ., Conclusions: Women with POF were more likely to exhibit ocular surface damage and symptoms of dry eye than age-matched controls. They were not, however, more likely to have reduced tear production. To our knowledge, this association between ocular surface disease and POF has not been previously reported. These data provide further evidence of the multifaceted role of sex hormones in the health and disease of the ocular surface.

Objectives: To compare conjunctival goblet cell numbers as well as epithelial turnover in patients with non-Sjögren syndrome--associated keratoconjunctivitis sicca (NSS-KCS) and those with SS-KCS before and after 6 months of treatment with topical cyclosporine A (CsA) ophthalmic emulsion., Methods: Conjunctival biopsy specimens from 16 patients with NSS-KCS and 12 with SS-KCS were obtained at baseline and after 6 months' therapy with CsA or vehicle alone. Conjunctival biopsy specimens were also obtained from 11 normal subjects. Periodic acid--Schiff staining determined the number of goblet cells present. Immunofluorescence microscopy for Ki-67 localization was used to evaluate the number of actively cycling cells., Results: Periodic acid--Schiff staining showed fewer goblet cells at baseline in both dry eye populations when compared with normal subjects (P<.001). After 6 months of CsA treatment, conjunctival biopsy specimens of both NSS-KCS and SS-KCS groups revealed an increase in goblet cells compared with baseline (P<.05). More Ki-67--positive cells were observed in NSS-KCS conjunctiva at baseline than in normal conjunctiva (P<.05) whereas numbers of these cells in SS-KCS conjunctiva were similar to normal at baseline. After 6 months of CsA treatment, conjunctival biopsy specimens of NSS-KCS revealed a decrease in Ki-67--labeled cells compared with baseline (P<.001). In contrast, no substantial change was observed for CsA treatment in patients with SS-KCS., Conclusions: Treatment of dry eye syndrome for 6 months with topical CsA resulted in an increase in goblet cell numbers in patients with NSS-KCS and SS-KCS and a decrease in epithelial turnover in those with NSS-KCS. Reducing ocular surface inflammation might have an effect on the proliferative activity of the epithelium.

Objective: To evaluate the effects of cyclosporin A (CsA) on cytokine and/or collagen production, cell growth, and apoptosis in conjunctival fibroblast cultures., Methods: Fibroblast cultures derived from normal subjects and patients with vernal keratoconjunctivitis and pemphigoid were exposed to different concentrations of CsA for either 24 hours or 30 days. The effects were evaluated by the colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) test to assess cell proliferation, and by the measurement of procollagen I (PIP) and procollagen III (PIIIP) cytokines and total protein in culture medium. CsA-induced apoptosis was assessed by fluorescence-activated cell sorter analysis., Results: After 24 hours of exposure to doses of CsA of more than 10 microg/mL, cell proliferation and migration were significantly reduced. Cyclosporin A reduced PIP and interleukin 1 (IL-1) production in a dose-dependent manner. Interleukin 6 and IL-8 were increased by 10 microg/mL of CsA, whereas transforming growth factor beta, PIIIP, and total protein were unaffected. Cyclosporin A exposure induced apoptosis in a time- and dose-dependent manner. Long-term exposure to CsA reduced IL-6 but did not modify PIIIP production., Conclusion: Exposure to CsA directly modified fibroblast behavior., Clinical Relevance: Cyclosporin A ability to accelerate apoptosis in clinically fibrotic tissues may prove to be therapeutic and useful in hyperproliferative conjunctival disorders.

Benign lymphoid hyperplasia of the conjunctiva occurs infrequently in children, and its presentation, clinical course, and appropriate management are not well established. We describe 2 children with nasal conjunctival masses that on pathological examination demonstrated benign lymphoid hyperplasia. Local irradiation of residual tissue was deferred, and the lesions remained stable for 1 year in one case and for 3 1/2 years in the other case. No systemic involvement had occurred. Although the natural history of extranodal lymphoid hyperplasia in children is poorly documented, most cases of nodal lymphoid hyperplasia in children are at very low risk of malignant transformation. Careful observation for local and systemic disease is indicated for ocular adnexal lymphoid hyperplasia in children until a more complete understanding of its natural history is available.

Objective: To evaluate the effects of topical ocular drugs with histamine H1-antagonist activity on histamine-stimulated phosphatidylinositol turnover and interleukin (IL) 6 and IL-8 secretion from human conjunctival epithelial cells., Methods: Primary human conjunctival epithelial cell cultures were stimulated with histamine in the presence or absence of test drugs. Phosphatidylinositol turnover was quantified by ion exchange chromatography and cytokine content of supernatants by enzyme-linked immunosorbent assay., Results: Antazoline hydrochloride, emedastine difumarate, levocabastine hydrochloride, olopatadine hydrochloride, and pheniramine maleate attenuated histamine-stimulated phosphatidylinositol turnover and IL-6 and IL-8 secretion. Emedastine was the most potent in ligand binding, phosphatidylinositol turnover, and IL-6 secretion, with dissociation constant and 50% inhibitory concentrations of 1-3 nmol/L. Olopatadine, antazoline, and pheniramine exhibited similar H1-binding affinities (32-39 nmol/L). However, olopatadine was approximately 10-fold more potent as an inhibitor of cytokine secretion (50% inhibitory concentration, 1.7-5.5 nmol/L) than predicted from binding data, while antazoline and pheniramine were far less potent (20- to 140-fold) in functional assays. Levocabastine (dissociation constant, 52.6 nmol/L) exhibited greater functional activity (50% inhibitory concentration, 8-25 nmol/L) than either antazoline or pheniramine., Conclusions: Histamine-stimulated phosphatidylinositol turnover and cytokine secretion by human conjunctival epithelial cells are attenuated by compounds with H1-antagonist activity. However, antihistaminic potency alone does not predict anti-inflammatory potential. Olopatadine, emedastine, and levocabastine were notably more potent than pheniramine and antazoline., Clinical Relevance: Selected topical ocular drugs with antihistaminic activity may offer therapeutic advantages to patients with allergic conjunctivitis by inhibiting proinflammatory cytokine secretion from human conjunctival epithelial cells.

Objective: To characterize the expression and regulation of conjunctival mast cell surface receptors important in allergic inflammation., Methods: Mast cells were isolated from human conjunctival tissues of cadavers. Mast cell surface markers were identified using flow cytometry with antibodies to IgE, Fc epsilonRI, c-kit, and intercellular adhesion molecule-1 (ICAM-1). We evaluated the effect of 24-hour tumor necrosis factor alpha (TNF-alpha) or interleukin 4 (IL-4) incubation on the expression of mast cell c-kit, ICAM-1, and surface-bound IgE., Results: Staining of mast cells (c-kit and/or tryptase positive) yielded positive results for all of the variables measured. The intensity of mast cell c-kit staining increased with TNF-alpha incubation, but decreased below that of unstimulated mast cells when incubated with IL-4. Anti-ICAM-1 and anti-IgE staining were increased over that of unstimulated cells when incubated with TNF-alpha or IL-4., Conclusions: In this model, TNF-alpha up-regulates mast cell surface receptors and cell-bound IgE. Interleukin 4 up-regulates mast cell ICAM-1 and cell-bound IgE, but down-regulates c-kit., Clinical Relevance: Conjunctival mast cells play a critical role in the pathogenesis of atopic ocular disease. Characterization of the expression and regulation of mast cell surface receptors is important to the development of potential novel treatments for ocular inflammation.

Objective: To assess azithromycin levels in human serum, aqueous humor, tear fluid, and conjunctival tissue specimens after administration of a single 1-g oral dose of azithromycin., Methods: Sixty patients undergoing cataract surgery were included in this analysis. Serum, aqueous, and tear specimens were collected 3, 6, and 12 hours and 1, 2, 3, and 4 days after azithromycin administration. Conjunctival tissue biopsy specimens were collected 1, 2, 3, 4, 6, 8, 10, 12, and 14 days after azithromycin administration. All specimens were subjected to analysis by high-performance liquid chromatography-mass spectrometry., Results: Azithromycin concentration ranges during the specified sampling times were as follows: serum, 21 to 974 ng/mL; tear, 82 to 2892 ng/mL; aqueous, 10 to 69 ng/mL; and conjunctival, 0.7 to 32 micrograms/g. Levels above the 90% minimal inhibitory concentration (MIC90) for Chlamydia trachomatis were detected after 4 days in all tear samples and after 14 days in all conjunctival tissue specimens following oral azithromycin administration., Conclusion: We demonstrated prolonged high levels of azithromycin in drug-targeted ocular tissue. Prolonged high concentrations of azithromycin in conjunctival tissue make this drug suitable for treatment of conjunctivitis caused by chlamydiae and other susceptible organisms.

Objective: To analyze the expression and cellular distribution of intercellular adhesion molecule 1, E-selectin (endothelial leukocyte adhesion molecule), vascular cell adhesion molecule 1, very late antigen 4, and lymphocyte function associated antigen 1 (LFA-1) in diseased and normal human sclera., Methods: Monoclonal antibodies to vascular cell adhesion molecule 1, very late antigen 4, intercellular adhesion molecule 1, LFA-1, and E-selectin were used to perform immunohistochemical staining on frozen sections of 16 cryopreserved human sclera specimens and 5 conjunctival specimens., Results: The normal human sclera did not express any of the adhesion molecules. The expression of LFA-1 was dramatic in all the scleral and conjunctival specimens on the inflammatory cells. Intercellular adhesion molecule 1, the ligand for LFA-1, was expressed in 7 of 12 scleral specimens. Furthermore, the expression of LFA-1 and intercellular adhesion molecule 1 were focally present in areas of inflammatory infiltrate. E-selectin expression was detected on the vascular endothelial cells in 8 of 12 patients. There was variable expression of vascular cell adhesion molecule 1 and very late antigen 4 in the inflamed sclera and conjunctiva., Conclusions: Our results demonstrate the presence of LFA-1 in the sclera and in the conjunctiva of patients with scleritis. Variable expression of other leukocyte adhesion molecules was noted in the sclera and the conjunctiva of these patients.

Objective: To determine the incidence of focal, late-onset, conjunctival bleb leaks after glaucoma filtering surgery., Design: Prospective, cross-sectional analysis., Setting: Tertiary care outpatient referral center., Patients: Consecutive patients who underwent glaucoma filtering surgery prior to June 1996 presenting for evaluation from September 2,1996, through November 15,1996. Five hundred twenty-five eyes of 525 consecutive patients were enrolled in the study., Intervention: Bleb height (elevated or flat), area (diffuse or localized), and wall thickness (thin, thick, or encapsulated) were classified. Each bleb was tested for focal leakage using a moistened fluorescein strip, cobalt blue illumination, and slit-lamp biomicroscopy. Diffuse transconjunctival aqueous flow did not qualify as a focal leak., Main Outcome Measure: Seidel-positive aqueous leakage., Results: Bleb leakage occurred in 14 eyes following trabeculectomy (mitomycin C treatment, 10 eyes; 5-fluorouracil treatment, 3 eyes; no antifibrosis agent, 1 eye) and in 1 eye following combined cataract and glaucoma surgery with adjunctive mitomycin C therapy. Bleb leakage occurred more frequently in eyes that received mitomycin C (10 [3.7%] of 273 eyes) than 5-fluorouracil (3 [1.4%] of 213 eyes) or no antifibrosis agent (1 [2.6%] of 39 eyes), using Kaplan-Meier estimates (P=.008, log-rank test). Conjunctival blebs were significantly thinner after trabeculectomy with mitomycin C than with 5-fluorouracil (P=.001). Bleb wall thickness was greater following combined cataract and glaucoma surgery than following trabeculectomy alone (P=.008). Age (P=.84), sex (P=.68), race (P=.77), duration of mitomycin C exposure (P=.62), number of antiglaucoma medications (P=.16), and total 5-fluorouracil dose (P=.85) were not associated with late-onset leaks., Conclusions: The risk of late-onset focal bleb leakage increases following trabeculectomy with mitomycin C therapy. Late leakage after combined cataract and glaucoma surgery is infrequent.

We studied the inhibitory effect of topical 1% indomethacin aqueous suspension on prostaglandin E2 in 20 conjunctivae extracted during cataract surgery and 16 corneas extracted during penetrating keratoplasty. Thirty minutes before each surgical procedure, indomethacin or placebo (0.9% sodium chloride solution) was instilled into the eye to be operated on in a masked fashion for conjunctiva and selectively for cornea. The excised corneal buttons and conjunctival tissue were homogenized and assayed for prostaglandin E2 concentration using radioimmunoassay. The levels of prostaglandin E2 in the conjunctiva significantly decreased (P less than .0033) after administration of indomethacin (203.7 +/- 219.5 pg/mg) compared with the placebo-treated tissues (849.8 +/- 564.1 pg/mg). The levels of prostaglandin E2 in the corneas significantly decreased (P less than .0157) after administration of indomethacin (522.5 +/- 570.2 pg in the cornea) compared with the placebo-treated tissues (1636.3 +/- 1344.1 pg in the cornea). Indomethacin may serve as a potent anti-inflammatory agent in conjunctival and corneal inflammatory disorders.

We correlated histologic findings concerning conjunctival goblet cells with findings concerning the mucous membrane surfaces of the conjunctiva. Ten biopsy specimens were obtained from the upper tarsal conjunctiva and ten from the perilimbal bulbar conjunctiva. Changes in goblet cells just before and during secretion included changes in surface cell diameter, evagination of the apical surface of the cell membrane, decrease in the number of microvilli, and alteration in the arrangement and morphologic characteristics of microvilli. Changes seen by light and transmission electron microscopy correlated with those observed by scanning electron microscopy. Goblet cells were often associated with crypt openings; some crypts served as conduits for the secretion of mucus from underlying goblet cells. This study supports the hypothesis that crypts in the conjunctiva seen by scanning electron microscopy are often associated with goblet cells and their mucus secretion.

Xeroderma pigmentosum (XP) is an autosomal recessive disease with tumor formation on sun-exposed areas of the skin and eyes. Cells from most XP patients are deficient in repairing DNA damaged by ultraviolet (UV) light as shown by a reduced rate of tritiated thymidine (3HTdR) incorporation during their DNA repair synthesis. We have studied such repair synthesis in conjunctival cells from an XP patient with a conjunctival epithelioma and from normal cadaver conjunctiva. Cultured conjunctival cells were irradiated with UV light and then incubated with 3HTdR. Autoradiograms were prepared and showed that UV radiation induced a considerably slower rate of DNA repair synthesis in the XP cells than in normal cells. Many of the ocular abnormalities of XP, including tumor formation, may be the result of this defective DNA repair process.

Corneal clouding is one of the manifestations of Tangier disease, an inherited disorder in which cholesterol-rich lipids are deposited in various tissues of the body. The cause of the corneal clouding is unknown. This study documents the clinical course and conjunctival biopsy findings of a 60-year-old man who was one of the earliest patients to be recognized with Tangier disease and in whom progressive corneal clouding developed in adult life. Noteworthy in the biopsy specimens were birefringent lipid particles that were predominantly present in degenerating pericytes of the conjunctival vessels.

A prospective, randomized study evaluated the effects of unilateral, topical 1% apraclonidine hydrochloride on conjunctival oxygen tension in 10 normal volunteers using a conjunctival oxygen monitor. Conjunctival oxygen tension and intraocular pressure were measured prior to instillation of apraclonidine, then at 1, 3, and 5 hours. Apraclonidine was found to significantly decrease conjunctival oxygen tension 76% at 1 hour compared with baseline values. At 3 hours conjunctival oxygen tension was decreased to 56% and at 5 hours to 10% of baseline. Intraocular pressure was lowered maximally at 3 hours to 40% of baseline measurements in the treated eye. The contralateral eye had a small decrease in conjunctival oxygen tension and intraocular pressure that was not statistically significant. Lid retraction and conjunctival blanching were noted to occur maximally between 1 and 3 hours. Further study is needed to determine if the apraclonidine-induced conjunctival hypoxia noted in this study has clinical significance for ocular blood flow, particularly in patients with glaucoma.

We have compared the effects of nonsteroidal anti-inflammatory agents (NSAIDs) indomethacin, flurbiprofen, and aspirin administered either intraperitoneally (IP) or topically on rabbit conjunctival and anterior uveal prostaglandin (PG) synthesis. Doses of IP flurbiprofen and aspirin at or above 50 mg/kg almost completely inhibited PG synthesis in the conjunctiva and anterior uvea. Indomethacin at doses up to 100 mg/kg only partially inhibited PG synthesis in rabbit anterior uvea. Although IP flurbiprofen and aspirin were better than IP indomethacin in inhibiting PG synthesis in anterior uvea, 0.5% topical aspirin almost completely inhibited PG synthesis in the conjunctiva and anterior uvea, but 0.5% flurbiprofen and 0.5% indomethacin did not. A lower topical dose (0.01%) of aspirin prevented the production of PGs in the anterior uvea.

Five milligrams of fluorouracil was injected subconjunctivally in the rabbit eye, and its concentration changes in the cornea, aqueous humor, and conjunctiva and sclera (both at the injection site and 180 degrees away from it) were determined by microbiological assay. The fluorouracil concentrations in the cornea and the aqueous humor averaged about 20 micrograms/g at one hour. The former decreased to 0.5 micrograms/g at 24 hours, while the latter was 0.03 micrograms/g at ten hours or later. At five hours, the fluorouracil concentrations in the conjunctiva and sclera at the injection site were similar to those 180 degrees away from it, both averaging about 2 micrograms/g. They decreased to 0.3 through 0.9 micrograms/g at 24 hours. The fluorouracil concentrations in the cornea, conjunctiva, and sclera at 24 hours were still above the reported 50% inhibition levels for the cultured conjunctival fibroblast. These findings have potential clinical implications for the safe use of fluorouracil.

Although the diagnosis of pseudoexfoliation syndrome (PXS) is made by observation of pseudoexfoliation material (PXM) deposited on the anterior lens capsule, there are several additional signs related to pigment dispersion that are often present both in eyes with PXS and fellow eyes in unilateral cases. On the basis of the presence of these signs, we have categorized a group of patients as "pseudoexfoliation suspects" who demonstrated no evidence of PXM on the lens capsule in either eye. As PXM has previously been identified in the conjunctiva of affected eyes as well as fellow eyes in unilateral cases, we hypothesized that it could also be present in PXS suspect eyes. Inferior bulbar conjunctival biopsies were performed on four eyes with PXS, five fellow eyes, and 23 PXS suspect eyes, and specimens were examined by transmission electron microscopy. In eight PXS suspect eyes biopsy specimens demonstrated PXM, suggesting that PXS is more prevalent and possibly responsible for a greater proportion of glaucoma than previously suspected.

The condition of a 4-year-old white girl of Ashkenazi Jewish parents was diagnosed as mucolipidosis IV on the basis of marked corneal clouding and severe psychomotor retardation, in the absence of facial-skeletal dysplasia or abnormal mucopolysacchariduria. The results of histochemical and ultrastructural studies of conjunctiva, skin, and corneal epithelium confirmed the combined storage of acid mucopolysaccharide and complex lipid substances. An unusual histopathologic feature of mucolipidosis IV is the predisposition of extreme storage involvement of corneal epithelial cells with relative sparing of the keratocytes, which is a finding of potential therapeutic implication. In addition, application of electron microscopic study of cultured amniotic cells and conjunctival biopsy specimens to assess for the parents the mother's subsequent pregnancy additional emphasizes the value of ultrastructural studies in the diagnosis of lysosomal storage disease.