Lipid-protein constituents of human corneal arcus.

Extracts of fresh senile human peripheral cornea with varying degrees of arcus were prepared by soaking minced tissue in buffered saline/EDTA. Apolipoprotein B was, at most, an occasional feature of these extracts; interactions involving glycosaminoglycans were not evident; and the lipid composition, particularly of the cholesterol ester fraction, was also not consistent with a recent origin from plasma components and particularly form low density lipoprotein. Assuming this origin, substantial secondary changes must follow insudation, involving protein loss and lipid reesterification, as is described for lipid deposits forming intracellularly at other sites. The manner of these changes in deposit forming extracellularly in the avascular peripheral cornea is not clear.

Analysis of lipid deposits extracted from human macular and peripheral Bruch's membrane.

OBJECTIVE: Lipids in Bruch's membrane may affect the evolution of age-related macular disease. To determine whether these show differences in regional distribution, we analyzed lipid deposits in Bruch's membrane at macular and peripheral sites. METHODS: Thin-layer chromatography was used to measure different lipid classes extracted from macular and peripheral Bruch's membrane of 32 eye bank eyes. RESULTS: The quantity of lipid extracted was consistently higher in the macula than in the periphery of human eyes. The total from both sites and the difference between the sites increased with age. The extracted lipids consisted largely of phospholipids, triglycerides, fatty acids, and free cholesterol. There was little cholesterol ester. CONCLUSIONS: Accumulation of lipids with age appears to be greater in the central than in the peripheral region of the fundus, indicating that lesions in age-related macular degeneration and Bruch's membrane lipid deposits share a common spatial distribution. The composition is consistent with the lipids being of cellular origin.

Tear calcium levels and contact lens wear.

Hydrophilic contact lens spoliation can be associated with the deposition of calcium salts. The relationship between the concentration of calcium in tear fluid and contact lens use has been studied in healthy volunteers and in patients using various forms of hard and soft contact lenses, tear samples being collected with Schirmer strips and with lenses in situ. Methods are suggested for correcting data on calcium per strip to concentration when this cannot be directly determined. Further data were obtained by direct extraction of freshly removed soft contact lenses. The collected results indicate some variation in tear calcium concentrations, but values are unrelated to use of contact lens and type of lens fitted, and to the rates of tear flow with our procedures for collection. Local calcium concentrations are unlikely therefore to be a significant primary factor in soft contact lens spoliation, but the enlargement of the tear pool associated with the use of a soft contact lens does greatly increase the amount of calcium present, and this may be a factor in secondary deposition.

Electrophoretic assessment of aqueous and serum neurone-specific enolase in retinoblastoma and ocular malignant melanoma.

The isoenzyme pattern of enolase was examined in the aqueous humour and serum of patients with retinoblastoma (10 aqueous, 8 sera), malignant melanoma (4 aqueous, 25 sera), and normal subjects undergoing cataract surgery (25 aqueous, 30 sera). The assay we used allowed assessment of all three major isoenzymes, including the gamma gamma isoenzyme (neurone-specific enolase). No enolase was detectable in normal aqueous; alpha alpha isoenzyme was present in the aqueous of one patient with malignant melanoma, while aqueous from all patients with retinoblastoma contained both alpha alpha and gamma gamma. Normal serum contained only an alpha alpha band, while serum from patients with retinoblastoma contained alpha alpha, alpha gamma, and gamma gamma bands (7 sera, 87.5%), or alpha alpha only (1 patient, 12.5%). All sera from patients with malignant melanoma contained the alpha alpha band, with low levels of gamma gamma in 16 (60%). In a single patient with Coats's disease alpha alpha was present in the serum, but no enolase was detected in aqueous. Increased amounts of gamma-containing isoenzymes of enolase are found in both serum and aqueous from patients with retinoblastoma. In malignant melanoma there is often an increase in serum gamma gamma enolase. The assessment of aqueous and serum enolase patterns may be of value in the diagnosis of retinoblastoma and malignant melanoma.

Comparative intraocular penetration of topical and injected cefuroxime.

AIMS: The choice of a prophylactic antibiotic for cataract surgery is dependent on its antibacterial activity and tissue penetration. The influence of the route and timing of administration of cefuroxime on its intraocular concentrations was examined. METHODS: 120 patients were recruited before cataract surgery into a prospective trial to compare the anterior chamber concentration of cefuroxime at a fixed time after administration by three routes. In a further 110 patients, the interval before sampling was varied in order to permit an examination of the kinetics of penetration. In another 10 patients, cefuroxime was given topically at the completion of surgery to assess the effect of a corneal wound on aqueous penetration. Cefuroxime concentrations were measured by high performance liquid chromatography on 0.2 ml samples of aqueous aspirated from the anterior chamber. Mean aqueous concentrations of cefuroxime for each group were compared using Student's t test. RESULTS: After 25 mg cefuroxime, mean aqueous concentrations increased in the order forniceal (< 0.1 microgram/ml) < topical (0.18 microgram/ml) < subconjunctival (2.31 microgram/ml) when sampled 12-24 minutes after administration. Aqueous concentrations of cefuroxime reached a peak between 80 and 110 minutes after both forniceal and peribulbar injection but were still rising at this time after subconjunctival injection. Topical application of 12.5 mg cefuroxime to eyes with a 10 mm corneal wound resulted in a mean aqueous concentration of 9.34 micrograms/ml. CONCLUSION: In the intact eye, only sub-conjunctival injection resulted in clinically significant aqueous concentrations of cefuroxime (> 1 microgram/ml) between 12 and 24 minutes after administration. For all routes, maximal aqueous concentrations were delayed by at least 80 minutes from administration. In the presence of a corneal wound, high aqueous levels of cefuroxime were rapidly attained after topical application.

Lipid keratopathy and atheromatosis in an SPF laboratory rabbit colony attributable to diet.

Sporadic cases of corneal opacity occurred in an SPF rabbit colony fed on a diet containing 10% white fish-meal. Routine histological monitoring also revealed atheromatous lesions involving the intima of the aorta, coronary arteries and the arteries of other organs in 29 rabbits examined. Serum cholesterol levels above 10 mmol/l were found in 7 rabbits, including 4 affected with corneal lesions, which proved to be lipid keratopathy on histochemical and electron microscopical examination. The serum cholesterol level was found to be within normal range (below 2.27 mmol/l) in 10 rabbits reared entirely on a modified version of the diet, where the 10% fish-meal content was replaced with 10% grass-meal. No more ocular or atheromatous lesions have been found since the adoption of the new diet for the colony.

[Biochemical and histochemical analysis of age related lipid deposits in Bruch's membrane]. / Biochemische und histochemische Analyse altersabhängiger Lipidablagerungen in der Bruchschen Membran.

In histochemical studies the age-related deposition of different lipids in Bruch's membrane has been seen. This is important for the pathogeneses of age-related macular degeneration due to changes in diffusion characteristics. Because the specificity of histochemical results is limited, a correlation of histochemical lipid analysis with biochemical methods was performed. The macular Bruch's membrane of 27 donors (age 1-97 years) was analyzed. The results of this analysis demonstrated that the increasing histochemical staining for lipids in Bruch's membrane corresponded with an increasing amount of lipids extracted from Bruch's membrane in biochemical analysis. This lipid deposition progressed exponentially with age. However, a large variation in the type of lipid deposited could be observed, with predominant deposition of phospholipids in some eyes and of neutral lipids in others. Furthermore, the phospholipids extracted were differentiated into 50% phosphatidylcholine, 30% phosphatidylethanolamine and 20% phosphatidylinositol and phosphatidylserin. The deposition of lipids, especially neutral lipids, in Bruch's membrane may change its diffusion characteristics. This appears to be important for the pathogenesis of age-related pigment epithelium detachments. The composition of the phospholipids extracted from Bruch's membrane supports, furthermore, the concept that the material deposited is the result of metabolic limitations in the degradation of photoreceptor outer segment material.

Erythrocyte stearic to oleic acid ratio in patients with ocular melanoma.

The main characteristics of malignant cells include increased metabolism and capacity for division which are associated with high membrane fluidity. Membrane fluidity is in turn controlled by multiple factors which include the ratio of saturated to unsaturated fatty acids in the cell wall i.e. the Saturation Index (SI). The SI is known to be reduced in neoplastic cells as well as in the circulating erythrocytes of patients with some malignant tumours but not others. It has been suggested that the SI may be useful as a marker of malignancy.

Problems with corneal arcus.

Corneal arcus presents many puzzling features. The correlation between its incidence and serum lipid levels is poor and, using immunoelectrophoresis, we have only been able to identify low-density lipoprotein inconsistently in corneae containing this deposition. Infrared thermography has shown us that arcus commences in the warmest regions of the cornea. We have considered the possible relevance of our biochemical and thermographic findings to other problems with corneal arcus such as its irreversibility, anatomical distribution, and clear zone.

Correlation between biochemical composition and fluorescein binding of deposits in Bruch's membrane.

PURPOSE: The fluorescence of drusen during fluorescein angiography is believed to have important prognostic and pathogenetic implications in age-related maculopathy. It is believed that deposits containing predominantly neutral lipids would be hydrophobic, resulting in hypofluorescence on fluorescein angiography, while the presence of polar phospholipids would be indicated clinically by hyperfluorescence because of its hydrophilic properties. To identify the potential determinants of fluorescence of drusen and Bruch's membrane, a series of macular specimens from human donors older than 60 years of age was examined. No clinical information was available concerning any previous eye disease. METHODS: In vitro fluorescein binding was recorded microscopically, and the presence of fibronectin was sought by immunohistochemistry. The results were correlated with the proportions of phospholipids to neutral lipids identified by histochemical and biochemical studies. RESULTS: It was found that high content of neutral fats was associated with lack of both fluorescein binding and fibronectin, and, conversely, in those specimens with high proportions of phospholipids, fluorescein binding was strong and fibronectin was present. CONCLUSIONS: These observations support the central hypothesis concerning biophysical changes in Bruch's membrane with age and the potential importance of fluorescein angiography in the characterization of Bruch's membrane deposits.

Familial lecithin:cholesterol acyltransferase deficiency. Biochemistry of the cornea.

Opacification of the cornea from lipid accumulation is an early and characteristic feature of familial lecithin:cholesterol acyltransferase (LCAT) deficiency. Visual impairment in a female age 48 years led to keratoplasty and the first detailed analysis of cornea in this disorder. Multilaminar figures were present, and total lipid extracts were enriched with phospholipid and cholesterol; cholesteryl esters were reduced, and accounted for about 12% of the cholesterol. Linoleate C18:2 was the predominant residue in the cholesteryl ester fatty acid fraction, with a C18:1/18:2 ratio of 1:6.5. This ratio differs from that in normal cornea, and from that in plasma and in other tissue deposits in LCAT deficiency. Various disorders of the HDL/LCAT system in plasma can lead to corneal lipid accumulation and opacification. These disorders may share general defects of lipid clearance from the cornea, but this study of LCAT cornea indicates that the character of the accumulating lipid is significantly influenced by active local metabolism, irrespective of the defect in the HDL/LCAT system also present.

Correlation between lipids extracted from Bruch's membrane and age.

BACKGROUND: There is increasing circumstantial evidence that the chemical composition of the deposits in Bruch's membrane influences the clinical outcome in age-related macular disease. In particular, it has been postulated that deposition of neutral lipids in Bruch's membrane may cause hydrophobicity and predispose to detachment of the retinal pigment epithelium and cause functional loss. METHODS: Analysis of the lipid extracted from Bruch's membrane of eye bank eyes from donors of different ages has been undertaken using thin layer and gas chromatography. No clinical information was available concerning any previous eye disease. RESULTS: It is shown that the lipid extracted increases with the age of the donor, and that the total quantity and ratio of neutral fats to phospholipids varies widely from one specimen to another from donors older than 60 years of age. The ratios of the different phospholipids imply that they are not derived from blood. CONCLUSION: The results are compatible with the concept of hydrophobicity developing with age in Bruch's membrane.

Penetration of mercury from ophthalmic preservatives into the human eye.

Single corneas of nine patients about to undergo full-thickness corneal keratoplasty were exposed to thiomersal by application of a hydrophilic gel contact lens preserved with this mercurial agent. These lenses were removed at operation 4 h later, and the lenses, corneal buttons, and samples of aqueous were taken for determination of total mercury by wet digestion and flameless atomic absorption. The contact lenses contained little residual mercury, but mercury was readily detected in the corneal buttons and in the samples of aqueous over the range of 1--60 ng/ml. Five control samples of aqueous obtained without exposure to mercurial preservatives contained less than 1 ng/ml mercury. Although specific effects on ocular function are not described, the use of these preservatives is associated with penetration of mercury into the aqueous in concentration similar to those reported with systemic poisoning by organic mercurials.