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To investigate the oxidative status in lens epithelial cells of patients with pseudoexfoliation (PEX) syndrome. Lens capsule 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 disulphide (GSSG), protein concentration, and lipid peroxidation. Concentrations of GSH, GSSG, lipid peroxidation, and protein concentration were determined by specific fluorescent assays. This study shows a 2.2- and 2.0-fold decrease in GSH and GSSG levels, respectively, in PEX lens epithelial lens compared with non-PEX lens epithelial cells, as well as a 2.5-fold increase in lipid peroxidation product malondialdehyde (MDA) levels. The increased MDA and decreased GSH levels indicate high oxidative stress. On the other hand, GSSG usually increases in cases of high-oxidative stress, but this is not always the case, as it may not always accumulate in cells. Our findings suggest a role for oxidative stress in the pathogenesis and the progression of PEX 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 o...

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-Grumwald-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.

Staphylococcus epidermidis is an important cause of bacterial keratitis. Certain S. epidermidis strains produce an extracellular slime layer rich in an acidic polysaccharide with a molecular size of 20 kDa (20-kDa PS). We have demonstrated that the level of 20-kDa PS-specific antibodies significantly rises after establishment of slime-producing S. epidermidis bacteraemia and, furthermore, that rabbit polyclonal antibodies to 20-kDa PS opsonize cells of slime-producing S. epidermidis to a great degree and promote their clearance by polymorphonuclear cells (Arch. Biochem. Biophys. 342 (1997) 389; J. Pharm. Biomed. Anal. 22 (2000) 1029). The purpose of this study was to examine the protective and therapeutic effects both of active immunization, using 20-kDa PS as antigen, and of passive administration of specific antibodies towards the 20-kDa PS in a rabbit keratitis model. For active immunization, 20 rabbits were subcutaneously immunized with 20-kDa PS, whereas for passive immunization specific polyclonal IgG antibodies against 20-kDa PS were administered to 20 rabbits 1 day before induction of infection. Clinical observations were made weekly for 1 month and levels of 20-kDa PS antibodies in serum and aqueous humor in both immunization groups were determined by an enzyme immunoassay. The levels of specific anti-20-kDa PS IgG in serum and aqueous humor following either active or passive immunization were significantly higher as compared with control groups (P

Malondialdehyde (MDA) is considered as the most important marker for monitoring lipid peroxidation, which is strongly associated with the development of serious diseases in adults and premature neonates. In this paper we report a method for determination of free MDA in human plasma using capillary zone electrophoresis. MDA was separated and determined as conjugate with tetrabutylammonium hydrogen sulphate (TBAS). Analysis was performed using 20 mM borare, pH = 9.3, as operating buffer and detection of the MDA-TBAS adduct at 267 nm. The method has a linear range up to 80 microM with a detection limit of 0.2 microM. The method was applied to the analysis of MDA in plasma of healthy adults, normal-gestation infants and of preterm neonates. Plasma proteins were successfully removed following centrifugation through a centricon-3 membrane. Results showed that the method can be easily and accurate applied for the determination of MDA in human plasma and that the level of MDA in pretern neonates is significantly higher (p

Objective: To investigate whether maternal smoking during pregnancy causes retinal abnormalities in the newborn. Study design: One hundred sixty-two neonates of smoking mothers and 162 matched neonates of nonsmoking mothers (112 appropriate for gestational age [AGA], 30 small for gestational age [SGA], 20 large for gestational age [LGA] in each group) were studied. Results: Retinal arterial narrowing and straightening (RANS) was observed in 52 and 10 eyes of the newborns of smoking and nonsmoking mothers, respectively ( P P P = .0007) in association with elevated hematocrit and RVDT, whereas no intraretinal hemorrhages were found when RANS was present. All retinal abnormalities resolved by 6 months in infants of smoking mothers and by 2 months in infants of nonsmoking mothers. Conclusions: Maternal smoking during pregnancy causes increased frequency of RANS, RVDT, and intraretinal hemorrhages; but these retinal abnormalities resolve by 6 months of age.

The aim of this study was the development of a capillary electrophoretic method for the determination of the levels of the selective alpha(2)-adrenergic receptor agonist brimonidine in aqueous humor of the eye and blood sera and their relation to its efficacy in reducing the intraocular pressure (IOP). Analysis of brimonidine was performed by capillary zone electrophoresis using 20 mM borate, pH 9.3, as operating buffer and detection at 255 nm. Brimonidine levels were determined in aqueous humor and blood sera from seven patients admitted for cataract extraction following ocular administration of the ophthalmic Alphagantrade mark solution. Levels of brimonidine and IOP values were recorded for a 24 h period. Alphagantrade mark administration resulted in a significant reduction of IOP, from within 30 min up to 4-5 h, whereafter a stepwise increase was recorded until 24 h, where mean IOP value returned to that before administration. The IOP reduction was related to the levels of brimonidine in aqueous humor, where maximal levels (80-100%) were obtained within 1-3 h. A 50% amount of the solution was determined after 4-5 h, whereas it reached the minimum level after 12 h. Serum levels reached maximum within 3-4 h, a 50% reduction was recorded in 12 h and minimum level in 24 h. It is concluded that brimonidine administration may significantly reduce IOP in patients when its level is maintained >/=50% of the maximum present in aqueous humor, i.e within a 4-6 h period. Since at this time the level of brimonidine in blood serum has reached maximum value, administration of brimonidine every 6 h may be used to obtain adequate brimonidine levels to maintain a constantly lowered IOP.

To develop scleral controlled-release-systems of triamcinolone acetonide (TA) based on biodegradable poly(lactide) (PLA).(1) PLA microspheres containing TA were prepared by a single or double emulsification-solvent evaporation method. Morphology, size, effect of drug input and method of microsphere preparation on drug loading, and in vitro TA release of the microspheres were investigated. (2) Mini-tablets consisting of blank PLA-microspheres and TA (weight ratios of 1:1, 2:1, and 4:1, respectively) were developed and their release profile in vitro was evaluated. (3) The in vitro transscleral diffusion profile was evaluated by placing a PLA-TA (1:1) tablet in a donor chamber and measuring the TA concentration in a receptor chamber. Donor and receptor chambers were separated by rabbit sclera. (4) Two cadaver rabbit eyes received a 1:1 PLA-TA tablet episclerally, which was covered by a scleral patch. TA aqueous humor and vitreous concentrations were measured 5, 10, and 20 days post implantation.(1) Microsphere average size was 2 μm. The double emulsification method and increasing drug input led to an increase in drug loading and encapsulation. Sustained release of TA over several days from the microspheres in vitro was observed, with the rate of release being affected by their TA content. (2) TA exhibited sustained release profile from the PLA-TA tablets, with the rate of release being affected by the PLA:TA ratio. (3) TA could slowly cross the sclera tissue in vitro, with approximately 21% of the drug loaded in the donor compartment being diffused through the sclera in 45 days. (4) Following scleral administration of the PLA-TA mini-tablets, TA accumulated in the vitreous and aqueous humor of cadaver eyes.The PLA-TA microspheres and mini-tablets appear promising for the controlled transscleral delivery of TA and justify further investigation.

A variety of syndromes leading to hemosiderosis in men cause ocular lesions. The purpose of the present study was to determine the distribution of iron and of transferrin in the eyes and lacrimal glands of rats in experimental hemosiderosis, so as to achieve a better understanding of the formation of the ocular lesions observed in patients with advanced hemosiderosis. In order to achieve hemosiderosis the rats were fed 3% (w/w) carbonyl iron or received i.p. or i.v. polymaltose iron. Hemosiderin deposits were detected in macrophages lying in the interlobular connective tissue of lacrimal glands, in the interstitial connective tissue of the choroid, in the ciliary body, in the iris and extracellularly in the sclera in all animals that received iron i.v. Also, scanty hemosiderin laden macrophages were found to a lesser degree in interstitial connective tissue of the choroid and in the interlobular connective tissue of lacrimal glands in animals that received iron i.p. No iron deposits were detected in the eye and lacrimal glands of control rats and in rats that were on an iron enriched diet. No transferrin was detected in the eye and in the lacrimal glands, neither in the control rats nor in the rats that received iron. Experimental iron overload leads to increased iron deposition in tissues of the eye and lacrimal glands, whereas no transferrin could be detected in the aforementioned organs.

Staphylococcus epidermidis is a leading cause of bacterial keratitis associated with corneal damage. Corneal integrity is closely associated with matrix macromolecules, such as proteoglycans (PGs) and collagen. The aim of this study was to examine whether active immunization (AI) using a major immunogenic polysaccharide determinant of slime (20-kDa PS) as antigen, and passive immunization (PI) after administration of specific antibodies toward 20-kDa PS affect the distribution of PGs as well as corneal lesions in an experimental model of slime-producing S. epidermidis keratitis.For AI, seven rabbits were immunized with 20-kDa PS, whereas for PI, seven rabbits received specific antibodies against 20-kDa PS. Lesions were graded clinically for a 21-day period. Levels of 20-kDa PS antibodies in serum and aqueous humor in both immunization groups were determined by ELISA. The distribution of certain extracellular matrix PGs during corneal healing was analyzed immunohistochemically.Levels of specific anti-20-kDa PS antibodies in serum and aqueous humor obtained after either AI or PI were significantly higher as compared with those in the respective nonimmunized control groups (p0.001). Clinical grading showed that both AI and PI rabbits had a significantly less corneal damage as compared with infected nontreated rabbits. Immunohistochemical analyses for PGs exhibited significant differences to the wounded regions as compared with noninfected corneal tissue. Accumulation of keratan sulfate PGs and decorin was observed in the corneal stroma of infected rabbits and of heparan sulfate PGs around the new-formed vessels. This phenomenon was significantly reduced in immunized animals in accordance with macroscopically decreased corneal damage observed in these animals.Results of this study suggest a key role of 20-kDa PS and its antibodies as prophylactic and therapeutic agents in keratitis caused by slime-producing S. epidermidis.

Pseudoexfoliation syndrome (PEX) is an age-related condition, which may cause open-angle glaucoma and has increasing interest since it seems to affect additional human tissues, i.e., cardiovascular tissue, skin, and still lacks elucidated pathogenesis. Collagen type IX and HNK-1 epitope have been considered characteristic constituents of the aqueous humour of PEX patients, since their amounts were increased in PEX aqueous humour compared to normal eyes. Since it has been proposed that the initial manifestations of PEX syndrome occur in conjunctiva, the present study was undertaken to investigate the presence of the same antigens in tears of PEX patients and their possible use as the biochemical markers for early diagnosis. Tears of PEX patients and healthy individuals were subjected to western blotting analysis for various basement membrane components identified in aqueous humour. It was found that collagen type IX and HNK-1 epitope were present in tears, the amount of the former being increased 2.7 times compared to normal (P0.05), surprisingly high as compared with total protein or lysozyme activity in tears, which were found to be increased in PEX patients about 25% with no statistical differences (P approximately 0.4). The results suggest the possible use of tears' collagen type IX for the diagnosis of PEX syndrome.

The aim of this study was the development of a capillary electrophoretic method for the determination of the levels of the selective alpha(2)-adrenergic receptor agonist brimonidine in aqueous humor of the eye and blood sera and their relation to its efficacy in reducing the intraocular pressure (IOP). Analysis of brimonidine was performed by capillary zone electrophoresis using 20 mM borate, pH 9.3, as operating buffer and detection at 255 nm. Brimonidine levels were determined in aqueous humor and blood sera from seven patients admitted for cataract extraction following ocular administration of the ophthalmic Alphagantrade mark solution. Levels of brimonidine and IOP values were recorded for a 24 h period. Alphagantrade mark administration resulted in a significant reduction of IOP, from within 30 min up to 4-5 h, whereafter a stepwise increase was recorded until 24 h, where mean IOP value returned to that before administration. The IOP reduction was related to the levels of brimonidine in aqueous humor, where maximal levels (80-100%) were obtained within 1-3 h. A 50% amount of the solution was determined after 4-5 h, whereas it reached the minimum level after 12 h. Serum levels reached maximum within 3-4 h, a 50% reduction was recorded in 12 h and minimum level in 24 h. It is concluded that brimonidine administration may significantly reduce IOP in patients when its level is maintained/=50% of the maximum present in aqueous humor, i.e within a 4-6 h period. Since at this time the level of brimonidine in blood serum has reached maximum value, administration of brimonidine every 6 h may be used to obtain adequate brimonidine levels to maintain a constantly lowered IOP.

To investigate whether maternal smoking during pregnancy causes retinal abnormalities in the newborn.One hundred sixty-two neonates of smoking mothers and 162 matched neonates of nonsmoking mothers (112 appropriate for gestational age [AGA], 30 small for gestational age [SGA], 20 large for gestational age [LGA] in each group) were studied.Retinal arterial narrowing and straightening (RANS) was observed in 52 and 10 eyes of the newborns of smoking and nonsmoking mothers, respectively (P. 000001) in association with elevated blood pressure in the neonates. The frequency of RANS was more than 3-fold greater in the SGA neonates than in the AGA and LGA neonates of the smoking mothers. Retinal venous dilatation and tortuosity (RVDT) was found in 100 and 36 eyes of neonates of smoking and nonsmoking mothers, respectively (P.000001). The frequency of RVDT in the SGA neonates of the smoking mothers was 2.5-fold and 4.2-fold greater than in the AGA infants and the LGA infants, respectively. Also, intraretinal hemorrhages were found in 61 and 31 eyes of neonates of smoking and nonsmoking mothers, respectively (P =.0007) in association with elevated hematocrit and RVDT, whereas no intraretinal hemorrhages were found when RANS was present. All retinal abnormalities resolved by 6 months in infants of smoking mothers and by 2 months in infants of nonsmoking mothers.Maternal smoking during pregnancy causes increased frequency of RANS, RVDT, and intraretinal hemorrhages; but these retinal abnormalities resolve by 6 months of age.

To evaluate the efficacy of topical brimonidine tartrate 0.2% (Allergan, Irvine, CA) in the prophylactic treatment of acute intraocular pressure (IOP) rises following Nd:YAG laser posterior capsulotomy.This was a double-masked, randomized, placebo-controlled clinical study in 60 patients who underwent Nd:YAG laser posterior capsulotomy after extracapsular cataract extraction. Two doses of brimonidine or vehicle were administered before and after capsulotomy. Intraocular pressure was measured prior to commencing instillation and for 48 hours postoperatively.Following posterior capsulotomy, the brimonidine group showed a significant mean percent reduction in IOP, while the vehicle group showed a significant mean percent increase in IOP. At 48 hours, mean IOP of both groups had returned to pre-laser level. There were no differences between the 2 groups in the incidence of clinical adverse experiences.Two-dose brimonidine tartrate 0.2% effectively prevents acute IOP rises after Nd:YAG laser posterior capsulotomy.

Contrast sensitivity function after cataract extraction and intraocular lens implantation has been mainly correlated to the type or the material of the intraocular lens. Our purpose was to identify other possible factors, like posterior capsule, to contrast sensitivity alterations after cataract surgery, comparing patients operated for cataract by techniques that mainly differed on the posterior capsule's integrity. The intraocular lens implanted was either a posterior or an anterior chamber one, always monofocal and made of PMMA. We measured contrast sensitivity function at four spatial frequencies in two groups of operated individuals (group A and B) and in one group of healthy control individuals. Each group consisted of 42 eyes. Group A comprised eyes with intact, clear posterior capsule and posterior chamber monofocal intraocular lens. Group B comprised eyes with ruptured or removed posterior capsule and anterior chamber monofocal intraocular lens. Control group comprised healthy control eyes. A pair matched design was used to compare contrast sensitivity values among the individuals of the three groups. No statistically significant differences in contrast sensitivity values were found when group B patients were compared to healthy controls (p0.05). Patients of group A exhibited contrast sensitivity function impairment at intermediate and high spatial frequencies when compared to patients of group B (p0.05) and to controls (p0.01). It seems that intact posterior capsule provides inferior visual function, in spite of relatively good visual acuity and apparently satisfactory results.

The concentrations of hyaluronan and galactosaminoglycans -- i.e., chondroitin sulfate and dermatan sulfate -- were measured in the aqueous humor of the eye from patients with exfoliation syndrome and from healthy persons. The glycosaminoglycans/proteoglycans were almost completely precipitated (97%) with ethanol in the presence of dextran as carrier and, following enzymic digestion, hyaluronan and galactosaminoglycans, were quantitatively converted to delta4,5-disaccharides. Non-degraded heparan sulfate and proteins/glycoproteins were removed by ultrafiltration using a Centricon 3 membrane. Separation and determination of hyaluronan- and galactosaminoglycan-derived delta-disaccharides were performed by ion-suppression HPLC. For an accurate analysis in triplicate, as little as 50 microl of aqueous humor is required. Application of this method to the analysis of samples from six patients with exfoliation syndrome and three healthy persons showed that hyaluronan levels in patients (6.65-16.15 microg ml(-1)) were significantly higher (3-8 times) than in healthy persons (2.0-2.24 microg ml(-1)). There was no significant alteration in the galactosaminoglycan concentration. The obtained data open a new area in the deeper understanding of the exfoliation syndrome pathophysiology and in establishing a highly sensitivity and accurate HPLC method for its diagnosis and patient's follow-up.

This study presents results obtained after implantation of 25 diffractive multifocal intraocular lenses. The follow-up ranged from six to 12 months. Visual acuity for distance without correction was 0.5 or better in 72% of cases; for near, it was J1 to J3 in 84% of cases. Slight decentration of the lens and pupil size did not affect visual acuity.

We examined 29 patients with homozygous beta thalassemia. The mean age of the patients was 15.6 +/- 8.9 years. Twelve patients (mean age, 02.0 +/- 10.4 years) had one or more ocular abnormalities. Five patients had degeneration of the retinal pigment epithelium, one had lens opacities, two had lens opacities and degeneration of the retinal pigment epithelium, one had vascular abnormalities and degeneration of the retinal pigment epithelium, one had angioid streaks, lens opacities, and degeneration of the retinal pigment epithelium, and two had angioid streaks and degeneration of the retinal pigment epithelium. These abnormalities were observed in patients with both forms of beta thalassemia, major and intermedia. The frequency of the ocular abnormalities increased with age. The youngest patient with an ocular abnormality was 6 1/2 years old. There was no correlation between the abnormalities observed and the serum ferritin level, the mean hematocrit value, and the dose of deferoxamine given to the patients.

Chemical burns were produced using a 4 N NaOH solution in 21 rabbit corneas. The ultrastructural findings were examined under an electron microscope five minutes and 1, 2, 4, 8, 15, and 30 days after the burn. A morphologic disturbance of the basement membrane and its complexes was detected from the first day of the experiment. These findings suggest that the basement membrane plays an important role in the pathogenesis of corneal recurrent erosions and ulcer formation after chemical burns.