Glutathione and lipid peroxide changes in pseudoexfoliation syndrome.

PURPOSE: To investigate the oxidative status of the aqueous humor of patients with pseudoexfoliation (PEX) syndrome. METHODS: Aqueous humor samples obtained during cataract surgery of patients with PEX syndrome and normal age-matched control subjects were examined for changes in the levels of glutathione (GSH), glutathione disulfide (GSSG), and TBA reactive species (TBARS), products of lipid peroxidation. GSH, GSSG, and TBARS were determined by specific fluorescent assays. RESULTS: Compared to normal controls, PEX syndrome aqueous humor samples showed a decrease of up to 28% of GSH concentration, and GSSG was increased up to 23%. The ratio of GSH/GSSG was 1.7-fold decreased in PEX syndrome samples. TBARS levels were increased by 100% in the PEX aqueous humor samples as compared to the controls. CONCLUSIONS: High levels of GSSG and TBARS indicate high oxidative stress, as well as the decrease in the ratio of GSH/GSSG. Our findings suggest a role for oxidation stress in the pathogenesis and the progression of PEX syndrome.

Quantitative and qualitative analyses of transferrin in aqueous humor from patients with primary and secondary glaucomas.

By using a highly sensitive and specific radioimmunoassay and the slot-blot technique, transferrin was quantified in fresh samples of aqueous humor from patients with primary open-angle glaucoma (POAG, n = 36) or secondary glaucoma (SG, n = 18). The levels were compared with those in aqueous humor obtained from age-matched patients without glaucoma (n = 33) and in primary and secondary aqueous humor from normal dogs (n = 10) in which breakdown of the blood-aqueous barrier was induced experimentally. The concentration of transferrin in the aqueous humor of human control subjects ranged from 0.3-3.4 mg/dl (mean +/- standard deviation, 1.36 +/- 0.66 mg/dl); in POAG samples, from 0.34 to greater than 10 mg/dl (2.07 +/- 1.90 mg/dl); and in SG samples, from 0.29 to greater than 10 mg/dl (2.79 +/- 2.24 mg/dl). The level of transferrin in secondary aqueous humor samples obtained from dogs was as much as ninefold greater than that in primary aqueous humor. The protein concentration in the human aqueous humor samples was 11.87 +/- 4.47 mg/dl for control subjects, 62.11 +/- 56.74 mg/dl for patients with POAG, and 124.53 +/- 152.67 mg/dl for those with SG. In dogs, the protein levels were 7.97 +/- 3.12 mg/dl for primary aqueous humor and 191.9 +/- 149.8 mg/dl for secondary aqueous humor. A significant correlation (r = 0.744, P less than 0.01) was found between total protein and transferrin contents in the samples of aqueous humor from patients with glaucoma but not in the samples from age-matched control subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Percutaneous anaesthesia with a lignocaine-prilocaine cream (Emla) for eyelid skin surgery.

Epicutaneous application of the anaesthetic cream Emla (lignocaine and prilocaine), to induce percutaneous anaesthesia in 38 patients scheduled for eyelid skin surgery is presented. The cream was applied 60 to 90 minutes before operation. In 36 out of the 38 patients (94.8%) no supplementary analgesia was required. The pain during the procedure was rated either as no pain in 29 patients (76.3%), slight pain in five patients (13%), moderate pain in two patients (5.2%), and severe pain in two patients (5.2%). Percutaneous anaesthesia induced by Emla cream is a useful and painless alternative method for analgesia in eyelid skin surgery.

Alterations in conjunctival cytology and tear film dysfunction in patients with beta-thalassemia.

PURPOSE: Patients with beta-thalassemia (beta-tha) represent a group with lifelong transfusion-dependent anemias. This study aimed to describe the conjunctival changes and tear film parameters in these patients. METHODS: A total of 52 patients (104 eyes) with beta-tha major and 22 normal control subjects (44 eyes) were studied during 1999 through 2000. Tear film break-up time (BUT), Schirmer test, rose Bengal staining, and cytologic evaluation of the conjunctival epithelium were performed in all subjects. The Papanicolaou and May-Grümwald-Giemsa staining procedures were performed on all smears. Patients and control subjects were compared for tear function parameters and conjunctival changes. RESULTS: The BUT, Schirmer test, and rose Bengal staining values were significantly lower (P < 0.001) in beta-tha patients than in control subjects. Keratinized cells were observed in conjunctival samples in 41% of patients, with a decrease in the number of goblet cells per slide in 64% of patients. In 9% of beta-tha patients, there were a slightly greater number of inflammatory cells than in control eyes. CONCLUSION: Ocular surface disorder of these patients was characterized by goblet cell loss and conjunctival squamous metaplasia. Our findings were correlated positively with the variable age. Epithelial damage by toxic reaction and disorder of tear quality and quantity are implicated as important factors in the pathogenesis of the ocular surface disease in beta-tha patients.

Changes in HNK-1 epitope and collagen type IX in the aqueous humour of patients with pseudoexfoliation syndrome.

PURPOSE: To investigate alterations in the proteoglycan (PG) and glycosaminoglycan (GAG) content of the aqueous humour in patients with pseudoexfoliation syndrome (PEX). MATERIALS AND METHODS: Aqueous humor samples were obtained during cataract surgery from nineteen patients bearing PEX features and twenty-three age-matched normal controls. Protein and IgG were quantified densitometrically after their electrophoretic separation. Collagen type IX, 3-sulphoglucuronic acid (HNK-1 epitope), biglycan and heparan sulphate proteoglycans were detected in Western and dot blots by using specific monoclonal antibodies (MAbs). The immunochemical analysis was performed in native aqueous humour or after degradation of the glycosaminoglycans with chondroitinases. RESULTS: Degradation of the samples with chondroitinases ABC, AC and B revealed that, in the aqueous humour from PEX eyes, collagen type IX and biglycan had a more dermatan sulphate than did normal eyes. In addition, more HNK-1 epitope was observed in PEX eyes, which after similar enzymatic treatment was found to be located mainly in dermatan sulphate sequences. 3-sulphoglucuronic acid was a constituent of the GAG chains of the collagen type IX. We found that the electrophoretic mobility of the bands of collagen type IX and HNK-1 epitope was exactly the same in the aqueous humour of normal and PEX samples; both migrated as four bands at 120, 113, 92.6 and 56 kDa. The PGs bearing heparan sulphate were found only in normal samples. Other PGs were not detected. CONCLUSIONS: Because no significant difference was observed in the concentration of albumin and IgG in PEX and normal samples, the blood-aqueous barrier was probably not significantly compromised in PEX patients with cataract but without open-angle glaucoma. The results support the hypothesis that the pathogenesis of PEX can be linked to disturbed metabolism of GAGs and PGs.