Lower Tear Meniscus Measurements Using a New Anterior Segment Swept-Source Optical Coherence Tomography and Agreement With Fourier-Domain Optical Coherence Tomography.

PURPOSE: To assess intraobserver repeatability and interobserver and intersession reproducibility of lower tear meniscus height (LTMH) measurements obtained using a new anterior segment swept-source optical coherence tomography (SS-OCT) device. Agreement with Fourier-domain (FD) OCT (Spectralis) was also examined. METHODS: In an observational cross-sectional study, one eye of 29 healthy subjects was randomly imaged with both devices at our center. Two examiners then randomly measured the LTMH using the software's calipers. To assess intraobserver repeatability and interobserver and intersession reproducibility, within-subject standard deviation (Sw), test-retest repeatability, coefficients of variation (CoV), and intraclass correlation coefficients (ICCs) were calculated. Agreement between both devices was also determined in Bland-Altman plots. RESULTS: Mean LTMHs for SS-OCT and FD-OCT were 276.6 ± 87.6 and 280.3 ± 80 µm, respectively. Using the SS-OCT device, intraobserver CoV, interobserver CoV, and intersession CoV were found to be ≥16.9%, ≤7.2%, and ≤11.5%, respectively. ICCs for these parameters were ≤88%, ≥97%, and ≥94%, respectively. Bland-Altman analysis indicated poor agreement between SS-OCT and FD-OCT, and the correlation was low (CoV 34.5%, ICC 0.36). CONCLUSIONS: SS-OCT LTMH measurements showed excellent interobserver and intersession repeatability along with good intraobserver reproducibility. Agreement between the devices was poor.

Extended imaging depth to 12 mm for 1050-nm spectral domain optical coherence tomography for imaging the whole anterior segment of the human eye at 120-kHz A-scan rate.

ABSTRACT. We demonstrate a 1050-nm spectral domain optical coherence tomography (OCT) system with a 12 mm imaging depth in air, a 120 kHz A-scan rate and a 10 µm axial resolution for anterior-segment imaging of human eye, in which a new prototype InGaAs linescan camera with 2048 active-pixel photodiodes is employed to record OCT spectral interferograms in parallel. Combined with the full-range complex technique, we show that the system delivers comparable imaging performance to that of a swept-source OCT with similar system specifications.

Evaluation of blotchy pigments in the anterior chamber angle as a sign of angle closure.

BACKGROUND: Blotchy pigments in the anterior chamber (AC) angle are considered diagnostic of primary angle closure (PAC). But there are no reports either on the prevalence of blotchy pigments in AC angles or the validity of this sign. AIMS: To determine the prevalence of blotchy pigments in AC angles and to evaluate their relationship with glaucomatous optic neuropathy (GON) in eyes with occludable angles. SETTING AND DESIGN: Cross-sectional, comparative study. MATERIALS AND METHODS: Gonioscopy was performed in 1001 eyes of 526 subjects (245 eyes of 148 consecutive, occludable angle subjects and 756 eyes of 378 non-consecutive, open angle subjects), above 35 years of age. Quadrant-wise location of blotchy pigments was documented. STATISTICAL ANALYSIS: Odds of blotchy pigments in occludable angles against that in open angles were evaluated. Relationship of GON with blotchy pigments in occludable angle eyes was evaluated using a multivariate model. RESULTS: Prevalence of blotchy pigments in occludable angles was 28.6% (95% CI, 22.9-34.3) and in open angles was 4.7% (95% CI, 3.2-6.3). Blotchy pigments were more frequently seen in inferior (16%) and superior quadrants (15%) of occludable angles, and inferior quadrant of open angles (4%). Odds of superior quadrant blotchy pigments in occludable angles were 33 times that in open angles. GON was seen in 107 occludable angle eyes. Blotchy pigments were not significantly associated with GON (odds ratio = 0.5; P = 0.1). CONCLUSIONS: Blotchy pigments were seen in 28.6% of occludable angle eyes and 4.7% of open angles eyes. Presence of blotchy pigments in the superior quadrant is more common in occludable angles. Presence of GON in occludable angle eyes was not associated with blotchy pigments.

Bioanalytical method validation of rapamycin in ocular matrix by QTRAP LC-MS/MS: application to rabbit anterior tissue distribution by topical administration of rapamycin nanomicellar formulation.

A novel, fast and sensitive 3200 QTRAP LC-MS/MS method was validated for rapamycin analysis in the rabbit eye following 0.2% administration of nanomicellar eye drop formulation. The LC-MS/MS technique was developed with electrospray ionization (ESI) in positive mode. Rapamycin was extracted from individual eye tissues and fluids by a simple protein precipitation method. Samples were reconstituted in 200µL of 80% of acetonitrile in water containing 0.05% formic acid. Twenty microliter of the sample was injected on LC-MS/MS. Chromatographic separations was achieved on reversed phase C 8 Xterra column, 50mm×4.6mm, 5µm. Multiple reactions monitoring (MRM) transition m/z 936.6/409.3 for rapamycin and 734.4/576.5 for erythromycin were employed as internal standard. The calibration curves were linear r(2)>0.9998 over the concentration range from 2.3ng/mL to 1000.0ng/mL. Rapamycin was found to be stable in ocular tissue homogenates for 6weeks at a refrigerated -80°C and -20°C temperatures. Rapamycin concentration was found to be 2260.7±507.1 (mean±S.D.)ng/g tissue and 585.5±80.1 (mean±S.D.)ng/g tissue in the cornea and iris ciliary muscle, respectively. This method has two advantages. First, a volatile base was used in the extraction procedure, which is easy to evaporate and generate consistent results. Second, the sodium adduct is employed that was stable in non-ammoniated mobile phase. The method demonstrates that absorption of rapamycin by a topical application of 0.2% rapamycin nanomicellar formulation generates therapeutically effective concentrations in the anterior segment of the eye.

Transport and interaction of cosmetic product material within the ocular surface: beauty and the beastly symptoms of toxic tears.

Eye cosmetics such as mascara, eye shadow and eyeliner are used extensively to highlight the eyes, and are normally applied external to the ocular surface. Adverse reactions of cosmetics within the ocular surface include mild discomfort, eyelid dermatitis, pre-corneal tear film instability, and keratitis. These are attributed mainly to the preservative (benzalkonium chloride (BAC)) constituent of cosmetic product material (CPM). Transport of CPM from an external environment to any location on the ocular surface, essentially precedes the adverse interactions occurring at the location, and the control of these transport modes is therefore of clinical relevance. The inter-transport of CPM across the TF occurs due to both diffusion and drift processes. Diffusion of neutral species is driven by concentration gradients, and the drift of cationic BAC is influenced by the inherent electric field; determined by the distribution of the various ions secreted into the aqueous layer, and the negative glycocalyx charge at the mucin layer. In the presence of mucin deficiency, the corneal epithelium is exposed to invasion by both incident BAC and lipophilic species. The transport of cationic BAC across the TF may be controlled by regulating the secretion of various electrolytes at the lacrimal gland. This is of clinical significance in reducing corneal epithelial adverse effects. However, the risks of adverse effects at the corneal surface due to invasion by the lipophilic species remain. Patients with mucin deficiency, and especially those on eye ointment/drops medication, should be discouraged from using cosmetics in a way likely to contaminate the TF.

Detecting endotoxin contamination of ophthalmic viscosurgical devices: intracameral versus intravitreal assays in rabbits.

OBJECTIVE: To compare the sensitivities of intracameral and intravitreal assays in the rabbit model to determine the relative adequacy of these methods in detecting bacterial endotoxin contamination of ophthalmic viscosurgical devices (OVDs). DESIGN: Experimental, randomized animal study. PARTICIPANTS: Twenty New Zealand white rabbits. METHODS: Rabbits were randomized into 4 groups to receive a cohesive or a dispersive OVD via intracameral or intravitreal injection. All 40 treated eyes (10 eyes of 5 animals in each group) received bilateral injection of OVD spiked with bacterial endotoxin at 7.0 endotoxin units/ml. All eyes were evaluated by slit-lamp biomicroscopy for inflammatory response at 3, 6, 9, 24, 48, and 72 hours after exposure. Eyes that received intravitreal injection were also dilated at 24, 48, and 72 hours and were re-examined by slit-lamp biomicroscopy and by indirect ophthalmoscopy. MAIN OUTCOME MEASURES: Conjunctival inflammation, anterior chamber (AC) flare, cells and fibrin, vitreous haze and cells, iridal hyperemia, corneal clouding, lens opacities, and onset times. RESULTS: Intracamerally injected eyes frequently showed conjunctival congestion, AC cells and flare, iridal hyperemia, and fibrin within 6 hours. Up to 80% showed AC cells and flare at 9 hours, and up to 70% showed fibrin at 24 hours. These signs diminished within 48 hours. Fibrin and cells also were seen on the lens surface of most of the eyes. Intravitreally injected eyes showed no signs of inflammation within 24 hours, other than some conjunctival inflammation. After the 24-hour time point, in addition to some conjunctival inflammation, some other signs of inflammation were observed infrequently in the intravitreally injected eyes, including minor vitreous cell reaction in 2 eyes. Although there was 1 dispersive OVD-treated eye with cells and fibrin on the lens capsule at 48 hours, no aqueous cells or flare were seen in the AC of any intravitreally injected eyes at any time during the course of the study. CONCLUSIONS: The rabbit intravitreal assay, when limited to 72 hours, does not seem to have adequate sensitivity to detect endotoxin reliably in OVDs.

Optical fiber-coupled ocular spectrometer for measurement of drug concentration in the anterior eye--applications in pharmaceuticals research.

This paper describes in detail a novel optoelectronic system designed to measure drug absorption in the anterior segment of the eye following topical application of drug formulations. This minimally invasive measurement technique offers both a method for determining drug concentration in human eyes, and demonstrates an alternative to current testing processes in model animals, which require paracentesis of the anterior chamber of the eye. The optoelectronic technique can be used with formulations, which possess appropriate spectral characteristics, namely unique absorption or fluorescence spectra. Preliminary experiments using our measurement system have been performed in rabbit and man, where we have been successful in achieving the direct measurement of topically applied brimonidine, an alpha-2 agonist used in the treatment of glaucoma. This demonstrates the feasibility of performing real-time, in vivo testing of ophthalmic drug formulations in the eye of human test subjects. We further demonstrate the novel application of the optoelectronic system for detection of topically applied UV-absorbing compounds in rabbit cadaver eyes, with a view to evaluating potential ocular sunscreen formulations. In summary, this method can be applied for the rapid comparison of the penetration of different drug formulations into the anterior eye at greatly reduced cost and time.

Tissue distribution of dexamethasone in canine ocular compartments following topical application of dexamethasone-21-isonicotinate and oxytetracycline HCl.

OBJECTIVE: To study the dexamethasone (DXM) concentration at different time points in various compartments of the canine eye following topical application of DXM-21-isonicotinate and oxytetracycline hydrochloride ANIMALS STUDIED: Thirty dogs to be euthanized for reasons not related to this study were selected and their ocular health status evaluated. Selected animals were treated with DXM-oxytetracycline ointment and euthanized after 6, 11 or 16 h. PROCEDURE: The concentration of DXM was determined in the following compartments of the eye: third eyelid, cornea, aqueous humor, iris, lens, vitreous body and choroid/retina. The DXM concentration in the eye was measured by radioimmunoassay. The applied amount of DXM was 0.04 mg in 0.2 mL ointment. Dogs were treated once with Corti Biciron eye ointment (DXM-21-isonicotinate and oxytetracycline hydrochloride, S & K Pharma, Perl, Germany) and were euthanized 6, 11 and 16 h after treatment. RESULTS: At 6 h following topical application the mean DXM concentration was highest in the anterior structures of the eye (third eyelid: 18 ng/g, cornea: 36 ng/g). The concentration in the posterior structures was below detection level. A decreased DXM concentration in the anterior structures was measured 11 and 16 h after treatment. CONCLUSION: It could be demonstrated that therapeutically relevant concentrations of DXM after a single topical administration are only achieved in anterior structures of the eye. A dosing interval of 6-11 h is recommended to achieve therapeutic drug concentration in those structures. The posterior structures of the eye are not reached by topical administration.

Identification of unknown intraocular material after cataract surgery: evaluation of a potential cause of toxic anterior segment syndrome.

PURPOSE: To describe and identify unknown opaque material between the optic of an AR40 intraocular lens (IOL) injected with the Emerald Series implantation system (both AMO, Inc.) and the posterior capsule at the conclusion of routine phacoemulsification to prevent an outbreak of toxic anterior segment syndrome (TASS). SETTING: Ambulatory care center operating room, University of North Carolina Hospitals and Department of Ophthalmology, University of North Carolina School of Medicine at Chapel Hill, Chapel Hill, North Carolina, USA. METHODS: After coaxial phacoemulsification in multiple patients, opaque material was present between the optic of a posterior chamber IOL and the posterior capsule. Although there was no TASS, the material was removed from 2 eyes and analyzed with scanning electron microscopy (SEM) and x-ray microanalysis (XRM). Similarly, crystalline lens, Klenzyme (Steris Corp.), Viscoat (sodium hyaluronate 3.0%-chondroitin sulfate 4.0%), and Provisc (sodium hyaluronate 1.0%) were analyzed. RESULTS: On SEM, the material had an irregular undulating surface similar to that of Provisc. Viscoat and the crystalline lens had smoother surfaces. On XRM, the material contained sodium, chlorine, and calcium, like Viscoat and Provisc, and phosphorous and sulfur, like Viscoat. The material also contained silicone, magnesium, aluminum, titanium, iron, and zinc. Klenzyme had smaller peaks of sodium, chlorine, and calcium and a higher carbon background than the unknown material. CONCLUSIONS: The material was likely ophthalmic viscosurgical device that was chemically and structurally altered by the cleaning and sterilization process. The silicone and metallic elements were probably from the Emerald Series implantation system as the disposable cartridge is coated with silicone and the reusable injector is metal.

Ghrelin as a novel locally produced relaxing peptide of the iris sphincter and dilator muscles.

Ghrelin is a recently described acylated peptide, which works as a somatosecretagogue and has described effects on the smooth, skeletal and cardiac muscle. We examined the production and effects of ghrelin on relaxation of the iris muscles. Contractile effects of 1-5 human ghrelin (frGhr, 10(-9)-6 x 10(-5)M) and 1-5 human des-octanoyl-ghrelin (d-frGhr; 10(-9)-6 x 10(-5)M) were tested on iris rabbit sphincter (n=11 frGhr; n=7 d-frGhr), dilator (n=6 frGhr; n=6 d-frGhr) and rat sphincter (n=6 frGhr; n=8 d-frGhr) precontracted muscles. On rabbit sphincter the effect of frGhr was also tested in presence of: i) L-NA (10(-5)M; n=7); ii) indomethacin (10(-5)M; n=7); iii) DLys(3)GHRP6 (10(-4)M; n=6); and iv) apamin+carybdotoxin (10(-6)M; n=6). Furthermore, on rabbit dilator the effect of frGhr was tested in presence of DLys(3)GHRP6 (10(-4)M; n=7). Finally, ghrelin mRNA production was assessed by "in situ" hybridization in Wistar rat eyes (n=8). In all muscles, frGhr promoted a concentration-dependent relaxation, maximal at 6 x 10(-5)M, 1.5-3 min after its addition, decreasing tension by 34.1+/-12.1%, 25.8+/-4.8% and 52.1+/-10.3% in the rabbit sphincter, dilator and rat sphincter, respectively. In the rabbit sphincter the relaxing effects of frGhr were: (i) enhanced in presence of DLys(3)GHRP6 (118.1+/-21.1%); (ii) blunted by indomethacin; and (iii) not altered by apamin+carybdotoxin (36.4+/-14.4%) or L-NA (52.4+/-11.4%). Relaxing effects of d-frGhr in rabbit (43.3+/-5.2%) and rat (77.1+/-15.3%) sphincter muscles were similar to those of frGhr. In rabbit dilator muscle, d-frGhr did not significantly alter active tension and the relaxing effect of frGhr was blunted by GHSR-1a blockage. Ghrelin mRNA was identified in iris posterior epithelium. In conclusion, ghrelin is a novel, locally produced, relaxing agent of iris dilator and sphincter muscles, an effect that is mediated by GHSR-1a in the former, but not in the latter. Furthermore, in the sphincter it seems to be mediated by prostaglandins, but not by NO or K(Ca) channels.

Innervation of the porcine ciliary muscle and outflow region.

The porcine eye serves as a model to study various functions of the aqueous outflow system. To compare these data with the primate eye, a detailed investigation of the distribution of contractile properties and of the innervation of the outflow region was conducted in the porcine eye. In all quadrants of the anterior eye segment, elastic fibres connected the ciliary muscle (CM) with the well-developed scleral spur (ScS) and also partly with the corneoscleral trabecular meshwork (TM) and the loops of the collecting outflow channels. Immunohistochemistry with antibodies against smooth muscle alpha-actin revealed intense staining of the CM and some myofibroblasts in the ScS and outer TM. In addition to a few cholinergic and aminergic nerve fibres in the outflow region, numerous substance P- and calcitonin-gene related peptide-positive nerve fibres and nerve endings were found near the outflow loops of the porcine TM. Although the porcine CM serves rather as a tensor choroideae muscle than as a muscle for accommodation, the innervation and morphology of the collecting outflow channel loops and of the expanded TM between the ScS and the cornea showed close similarities to the primate eye.

Substance P and opioid peptidergic innervation of the anterior eye segment of the rat: an immunohistochemical study.

Recently discovered endogenous opioid peptides such as nociceptin are known to modulate neurotransmitter release of primary afferent neurons (especially substance P, SP) and they have also been demonstrated in peripheral nerve fibres. The aim of this study was to investigate the opioid peptidergic innervation of the anterior eye segment and to compare it with the innervation pattern of SP in order to shed light on the functional relationship between these peptides. Anterior eye segments of 20 rat eyes were cut in a tangential plane and the sections stained with antibodies against SP, nociceptin, nocistatin, endomorphin 1 and 2, leu-enkephalin and met-enkephalin. Sections of the spinal cord or brain were used as positive controls. Numerous SP-immunoreactive nerve fibres were found in the conjunctiva, cornea, episclera, trabecular meshwork, iris and ciliary body. A weak staining for met-enkephalin and leu-enkephalin could only be found in the iris and anteriormost ciliary body. Nerve fibres immunoreactive for nociceptin, nocistatin, and endomorphin 1 or 2 could not be detected in any part of the anterior eye segment. It is tempting to speculate that the opioid peptidergic innervation of the anterior ciliary body may play a role in the modulation of intraocular inflammation.

Distribution of syndecans 1-4 within the anterior segment of the human eye: expression of a variant syndecan-3 and matrix-associated syndecan-2.

Control of the actomyosin network plays a role in regulating the movement of aqueous humor through the anterior segment of the eye. Receptors that could control its activity are unknown. In this study, we show that all four members of the syndecan family, which can regulate the actomyosin network, are present within the anterior segment. In both sections of human anterior segments and cultures of human trabecular meshwork (HTM), Schlemm's canal (HSC) and the ciliary muscle (HCM) cells from the anterior segment, syndecans-3 and -4 were the predominant family members. They were widely distributed throughout the anterior segment. Syndecan-3 within the anterior segment was a novel, recently described variant 55 kDa form. Low levels of syndecans-1 and -2 were also observed in situ and in all three cultures. Their expression was weaker and more localized than that observed for syndecans-3 and -4. Staining for syndecan-1 in HCM cultures was variable. In HTM and HSC cultures, syndecan-2 also co-distributed with fibronectin, laminin and type IV collagen suggesting that it was shed and associated with the extracellular matrix. Western blots supported this idea and showed syndecan-2 ectodomains in lysates from anterior segments.

Localization of thrombomodulin in the anterior segment of the human eye.

PURPOSE: To localize thrombomodulin (TM) in the anterior segment of the human eye. TM is a vascular endothelial cell surface glycoprotein that acts as a cofactor for the thrombin-catalyzed activation of the anticoagulant protease zymogen, protein C. METHODS: Immunohistochemical methods were used to detect TM expression in corneal epithelial cells, the lens epithelial cells, and other cells in the anterior segment of the eye. The expression of TM was also examined in cultured human corneal epithelial cells. RESULTS: TM was expressed in corneal epithelial cells, corneal endothelial cells, and nonpigmented ciliary epithelial cells, which are in direct contact with the aqueous humor. TM was also expressed in cultured corneal epithelial cells and showed cofactor activity. The amount of the antigen in the cultured corneal cells was approximately one tenth of that in human umbilical vein endothelial cells, but its specific cofactor activity (75%) was comparable to that of TM in human umbilical vein endothelial cells. The trabecular meshwork and endothelial cells lining Schlemm's canal also showed positive staining for TM. CONCLUSIONS: The TM in the cells that are in contact with the aqueous humor appears to be involved in maintaining the fluidity of the aqueous humor. In contrast, TM in cells that are not in contact with the aqueous humor may function in regulating cell proliferation and/or differentiation.

Visual pigments, oil droplets, ocular media and cone photoreceptor distribution in two species of passerine bird: the blue tit (Parus caeruleus L.) and the blackbird (Turdus merula L.).

The spectral absorption characteristics of the retinal photoreceptors of the blue tit (Parus caeruleus) and blackbird (Turdus merula) were investigated using microspectrophotometry. The retinae of both species contained rods, double cones and four spectrally distinct types of single cone. Whilst the visual pigments and cone oil droplets in the other receptor types are very similar in both species, the wavelength of maximum sensitivity (lambda(max)) of long-wavelength-sensitive single and double cone visual pigment occurs at a shorter wavelength (557 nm) in the blackbird than in the blue tit (563 nm). Oil droplets located in the long-wavelength-sensitive-single cones of both species cut off wavelengths below 570-573 nm, theoretically shifting cone peak spectral sensitivity some 40 nm towards the long-wavelength end of the spectrum. This raises the possibility that the precise lambda(max) of the long-wavelength-sensitive visual pigment is optimised for the visual function of the double cones. The distribution of cone photoreceptors across the retina, determined using conventional light and fluorescence microscopy, also varies between the two species and may reflect differences in their visual ecology.

Distribution of ascorbate in the anterior bovine eye.

PURPOSE: To analyze the ascorbate distribution in the anterior eye wall to better understand the functional significance of this compound in the eye. METHOD: Ascorbic acid was determined by high-performance liquid chromatography using an LC-10 system (Shimadzu, Kyoto, Japan). Bovine eye samples were used. RESULTS: The highest ascorbate concentration was observed in the corneal epithelium, with significantly higher values in the central (1.56 mg/g) than in the peripheral (1.39 mg/g) area. The ascorbate content was similar in the corneal stroma (0.22 mg/g), the Descemet's membrane (DM)/endothelium (0.22 mg/g), and the aqueous humor (0.21 mg/ml). By comparison, the sclera (0.15 mg/g) and the conjunctiva (0.11 mg/g) showed lower values, as did the lacrimal gland (0.09 mg/g) and the serum (0.0008 mg/ml). CONCLUSIONS: (1) Peak ascorbate concentration was observed in the central corneal epithelium covering the pupillary area. This is compatible with the idea that the ascorbate may act as an UV filter shielding internal eye structures from radiation damage. (2) The ascorbate concentration in the corneal stroma and DM/endothelium was as high as in the aqueous humor, and it is suggested that the aqueous humor plays a key role in the distribution of ascorbate to the anterior eye wall.

Cytoskeleton and tissue origin in the anterior cynomolgus monkey eye.

We studied cytoskeletal proteins and other markers for embryologic origin in the outflow pathways of the aqueous humor, cornea, sclera, and ciliary muscle of the cynomolgus monkey. The corneal endothelium and trabecular cells stained with markers for vimentin, smooth muscle cell alpha-actin, F-actin, spectrin, vinculin, and talin. The endothelium of Schlemm's canal stained with markers for vimentin, spectrin, and F-actin. These results suggest that trabecular cells are a kind of myofibroblast and support the belief that the endothelial cells of Schlemm's canal are vascular in origin. Fibrillary staining with antibodies to vimentin, spectrin, neurofilament protein, and glial acid fibrillary protein was observed along and between the ciliary muscle cells. Cells in the deep sclera adjacent to the supraciliary space stained with antibodies to smooth muscle alpha-actin, alpha-vinculin, talin, and desmin. These cells may anchor ciliary muscle cells into the sclera or may be developmental remnants of ciliary muscle cells. Leu 19 immunoreactivity was found in the corneal endothelium, in all trabecular cells, in ciliary muscle cells, and in keratocytes and fibroblasts in the superficial part of the cornea and sclera. All of these cells are therefore likely to express neural cell adhesion molecules indicating neuroectodermal origin.

[An immunohistochemical study on glycosaminoglycans distribution in the trabeculum of congenital aniridia].

Distribution of glycosaminoglycans in the trabecular tissue was immunohistochemically investigated in 7 congenital aniridia eyes of 5 patients aged 0 to 43 days. Paraffin sections of each specimen were immunohistochemically stained with antibodies to chondoroitin (clone 1-B-5), chondoroitin-1-sulfate (2-B-6), chondoroitin-6-sulfate (3-B-3), dermatan sulfate (6-B-6), and keratan sulfate (5-D-4). The trabecular meshwork and Schlemm's canal in all eyes were absent or not well differentiated. The cornea, trabecular tissue, and iris stroma were negative for chondoroitin immunostaining but positive for chondoroitin-4-sulfate, chondoroitin-6-sulfate, dermatan sulfate and keratan sulfate immunostaining. In the normal anterior segment tissue keratan sulfate is present in the cornea, trabecular tissue, and iris at the fetal stage, and disappears from the iris at the neonatal stage. These findings suggest that the persistence and/or abnormal distribution of keratan sulfate in the anterior segment may play a role in the pathogenesis of congenital aniridia.