Specular microscopic study of X-ray-irradiated rabbit cornea.

PURPOSE: To investigate the influence of ionizing radiation on the corneal epithelium and endothelium of rabbit eyes. METHODS: Five healthy mature albino rabbits were unilaterally irradiated with 20 Gy of X-rays (250 kV, 12 mA). Slit-lamp biomicroscopic observation and specular microscopic examination of the corneal epithelium and endothelium were carried out before and 1, 4, 8, 12, 16, 20, 24, and 36 weeks after irradiation. We evaluated mean area of the superficial corneal epithelial cells, mean area and the percentage of hexagonal cells of the corneal endothelial cells, and corneal thickness. The statistical difference between the irradiated and control eyes was assessed using paired t-test. RESULTS: All animals developed cataract within 24 weeks. Slit-lamp biomicroscopy showed no apparent corneal abnormalities over the 36-week follow-up period. Specular microscopy revealed a significant enlargement of the superficial corneal epithelial cells from 4 to 12 weeks after irradiation (P<0.01), which disappeared at 16 weeks post-irradiation. Specular microscopy of the corneal endothelium showed enlargement and morphological alterations of the cells beginning 8 weeks after irradiation (P<0.05). These changes persisted throughout the study period. There were no statistically significant changes in corneal thickness. CONCLUSION: After X-ray radiation of 20 Gy, transient damage occurred in the corneal epithelium, while delayed and irreversible changes were seen in the endothelium.

Mirror image myopic anisometropia in two pairs of monozygotic twins.

Two sets of monozygotic twins with mirror image myopic anisometropia are reported. The first set were two boys aged 1 year 8 months. There was a right eye myopic anisometropia in one twin, and a left eye myopic anisometropia in the other. The differences in refractive power between both eyes were 11.6 and 7.6 dpt, respectively (spherical equivalent). The second set were two 6-year-old boys. The right eye had myopic anisometropia in one twin, while the left eye was affected in the other. The differences in refractive power between both eyes were 6.5 and 3.7 dpt, respectively (spherical equivalent). Exotropia was recognized in 3 cases. Previously only two sets of monozygotic twins with mirror image myopic anisometropia have been reported. Monozygotic twins with mirror image myopic anisometropia are extremely rare.

Factors associated with increased aqueous flare in psoriasis.

PURPOSE: To evaluate the changes in aqueous flare in psoriasis patients and to identify the factors that influence the level of aqueous flare. METHODS: We examined the 68 eyes of 34 psoriasis patients and the 68 eyes of 34 healthy subjects with a laser flare-cell meter. Complete dermatologic and ophthalmic examinations were performed on the psoriasis patients. RESULTS: Flare was significantly higher in psoriasis patients than in normal controls (P <.000l). The factors that increased flare significantly were age and Psoriasis Area and Severity Index. Flare was not significantly associated with sex, psoriasis type, duration of disease, and cyclosporin therapy. A flare increase was significantly correlated with serum total protein and immunoglobulin (Ig) A, but not with albumin, IgG, and IgM. CONCLUSIONS: Psoriasis patients, even without ocular symptoms, had slight damage to the blood-aqueous barrier. Multiple linear regression analysis showed that flare had the strongest correlation with the severity of psoriasis.

Ultrastructural fragility and type IV collagen abnormality of the anterior lens capsules in a patient with Alport syndrome.

PURPOSE: To investigate ultrastructural and immunohistological abnormalities of the lens capsules in a patient with Alport syndrome. METHODS: Two anterior lens capsules were obtained at phacoemulsification from a 43-year-old female patient with bilateral lenticonus who was affected by Alport syndrome. The right capsule was examined by transmission electron microscopy and the left capsule was stained with two monoclonal antibodies against the triple-helical domains of type IV collagen alpha2 and alpha5 chains. RESULTS: Numerous vertical dehiscences with many disrupted interdigitations were observed in the right anterior lens capsule. Decreased reactivity of monoclonal antibodies against type IV collagen alpha5 chain and normal reactivity against the alpha2 chain were shown in the left anterior lens capsule. CONCLUSION: The ultrastructural fragility of the anterior lens capsules in this patient with Alport syndrome appears to be associated with the abnormality of the type IV collagen molecules including the alpha5 chain.

Refractive changes in diabetic patients during intensive glycaemic control.

AIMS: To evaluate the clinical course and the characteristics of transient refractive error occurring during intensive glycaemic control of severe hyperglycaemia. METHODS: 28 eyes of patients with persistent diabetes were included in this prospective study. During the observation period, patients underwent general ophthalmological examination and A-mode scan ultrasonography was performed at each examination-at days 1, 3, and 7, and then once every week or every other week until recovery of hyperopia. RESULTS: A transient hyperopic change occurred in all patients receiving improved control after hyperglycaemia. Hyperopic change developed a mean of 3.4 (SD 2. 0) days after the onset of treatment, and reached a peak at 10.3 (6. 1) days, where the maximum hyperopic change in an eye was 1.47 (0. 87) D (range 0.50-3.75 D). Recovery of the previous refraction occurred between 14 and 84 days after the initial assessment. There was a positive correlation between the magnitude of the maximum hyperopic change and (1) the plasma glucose concentration on admission (p<0.01), (2) the HbA(1c) level on admission (p<0.005), (3) the daily rate of plasma glucose reduction over the first 7 days of treatment (p<0.001), (4) the number of days required for hyperopia to reach its peak (p<0.001), and (5) the number of days required for the development and resolution of hyperopic changes (p<0.0001). There was a negative correlation between the maximum hyperopic change of an eye and baseline value of refraction (p<0.01). During transient hyperopia, no significant changes were observed in the radius of the anterior corneal curvature, axial length, lens thickness, or depth of anterior chamber. CONCLUSIONS: The degree of transient hyperopia associated with rapid correction of hyperglycaemia is highly dependent on the rate of reduction of the plasma glucose level. A reduction of refractive index in intraocular tissues, especially in lens, appears to be responsible for this hyperopic change.

Corneal endothelial changes and aqueous flare intensity in pseudoexfoliation syndrome.

PURPOSE: To determine the changes of corneal endothelium in pseudoexfoliation (PEX) syndrome and the relations between their changes and aqueous flare intensity. METHODS: 26 eyes with PEX syndrome, 17 clinically unaffected fellow eyes and 27 normal age-matched eyes were studied. The corneal endothelium of the eyes was examined using a specular microscope, and the aqueous flare intensity of the eyes was measured using a laser flare cell meter. RESULTS: The corneal endothelial density significantly decreased both in the eyes with PEX and in the clinically unaffected fellow eyes compared to the normal control eyes (p < 0.001, p < 0.01). The acqueous flare intensity significantly increased in PEX syndrome (p < 0.01). There was a significantly inverse correlation between the corneal endothelial cell density and the aqueous flare intensity in PEX. CONCLUSION: A decrease in corneal endothelial cells was quantitatively identified in PEX syndrome. This change might have a relationship with a disorder of the blood-aqueous barrier in this syndrome.

[Vascular endothelial growth factor levels in the aqueous and serum in diabetic retinopathy with or without neovascular glaucoma].

We determined the levels of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) in the aqueous and serum in non-insulin dependent diabetic patients with proliferative retinopathy (n = 12) and neovascular glaucoma (n = 11). The aqueous levels of PEGF/VPF were significantly higher in both groups than in 10 diabetics without such complications. The levels were very high in patients with neovascular glaucoma, suggesting that VEGF/VPF is involved in the pathogenesis and progression of diabetic neovascular glaucoma. The serum levels were not significantly related to the presence or the stage of retinopathy. The findings suggest the possibility of treatment of neovascular glaucoma using anti- VRGF/VFP preparations.

[Quantitative evaluation of aqueous flare in psoriasis using a laser flare-cell meter].

We evaluated aqueous humor protein concentration in psoriasis using a laser flare-cell meter, which can quantify aqueous flare precisely and objectively. Psoriatic severity was evaluated on the basis of psoriasis area and severity index (PASI) score. Aqueous flare was measured in 40 eyes of 20 psoriasis patients (sixteen psoriasis vulgaris, three guttate psoriasis, and one psoriatic arthritis) and 28 eyes of 14 normal controls. Aqueous flare value was significantly higher in psoriatic patients than in normal controls (p < 0.01). There was no difference between psoriasis vulgaris and the other types of psoriasis. Aqueous flare value was higher in patients with psoriatic history longer than 10 years than in those with less than 10 years (p < 0.05), and also higher in patients with severe psoriasis (PASI score > 10) than in those with mild psoriasis (PASI score < 10) (p < 0.05). But no statistically significant differences in aqueous flare value were found among cyclosporin, etretinate, and psoralen ultra violet A therapies. These findings strongly suggest that patients suffering from psoriasis have slight damage of the blood-aqueous barrier even if they have no ocular symptoms, and that the degree of blood-aqueous barrier damage increases with time and severity of psoriasis.

[A specular microscopic study on irradiated rabbit corneas--report I. Early injury of the cornea].

To evaluate the effects of ionizing radiation on the cornea, one eye of healthy mature albino rabbits (n = 5) was irradiated with 20 Gy of X-rays (250 kV, 12 mA). Corneal observation was done with a slit-lamp biomicroscope and a specular microscope before irradiation, and follow-up observations were done at one week and 4 weeks after irradiation. Specular microscopic photographs were analyzed with a digitizer. Although slit-lamp biomicroscopy did not show any abnormal findings in the cornea, specular microscopy revealed a significant enlargement of the mean cell area of the superficial corneal epithelium at 4 weeks after irradiation. There were no significant changes in the corneal endothelium during this study. These results strongly suggest that high doses of X-ray irradiation could induce corneal epithelial injury at the early phase, even when by slit-lamp biomicroscopy shows no abnormal findings in the cornea.

Survey of risk factors for expulsive choroidal hemorrhage: case reports. Substantiation of the risk factors and their incidence.

To evaluate the interaction of risk factors for expulsive hemorrhage, we evaluated 3 cases of expulsive hemorrhage and 2 of acute intraoperative choroidal effusion that occurred at the University of Tsukuba Hospital over a 16-year-period and investigated the common findings in these 5 cases. We also selected randomly 500 cataractous cases operated at the University of Tsukuba Hospital and investigated the probabilities of risk factors for expulsive hemorrhage in these control cases. The 5 affected patients exhibited more than four risk factors of expulsive hemorrhage. The probability of more than four risk factors being involved in any cataract operation was approximately 0.33% (0.17 approximately 0.67%; 95% confidence interval). The incidence of expulsive hemorrhage at our hospital was approximately 0.2%, which is similar to the probability rate. Thus, we concluded that the development of expulsive hemorrhage was proved to be related to an accumulation of the risk factors for expulsive hemorrhage.

[The effect of subconjunctival injection of endothelin-1 on intraocular pressure in the rabbit].

Endothelin-1 (ET-1) is a potent vasoconstrictive peptide produced in part by vascular endothelial cells. In order to investigate its effects on the episcleral vascular system and intraocular pressure (IOP), we injected ET-1 (3, 10, 30, 100, 300, 1,000 pmol) into the subconjunctival space of rabbits and measured IOP with a manometer. Injection of a dose higher than 10 pmol caused a transient increase of IOP. Mean maximum elevation rate of IOP for each dose of ET-1 was 14.9 +/- 0.9%, (mean +/- standard error) 43.5% +/- 9.5%, 40.8 +/- 7.5%, 46.9 +/- 9.8%, and 84.1 +/- 22.6. Next, we injected 1,000 pmol into the subconjunctival space, and continuously measured IOP and ocular pulse pressure with a manometer. IOP increased rapidly after ET-1 injection. Maximal increase of IOP was observed at 22.7 +/- 9.2 min after ET-1 injection, and IOP decreased after the peak. The ocular pulse pressure increased with IOP elevation and decreased with the IOP reduction. We speculated that the transient elevation of IOP was caused by increase of aqueous outflow resistance, and the decrease of IOP was caused by decrease of aqueous outflow resistance and decrease of blood flow in the ciliary body and the choroid. This strongly suggests that subconjunctival injection of ET-1 could have a large effect on the episcleral vascular system, aqueous outflow, and blood flow in the ciliary body and the choroid.

Increase in choroidal blood flow in rabbits with endothelin-1 induced transient complete obstruction of retinal vessels.

BACKGROUND: In a previous paper, we reported that retinal blood flow (RBF) ceased immediately after injection of 1 nmol endothelin-1 (ET-1) and no recovery of RBF was detected for at least 50 min. In this study, we confirmed the same duration of RBF cessation and measured choroidal blood flow (CBF) for 180 min. METHODS: We measured CBF in a rabbit model of transient complete obstruction of retinal vessels induced by intravitreal injection of a high dose of ET-1, using the hydrogen clearance method. We also investigated the effects of intravitreal injection of ET-1 on intraocular pressure (IOP), blood pressure, pulse rate and blood gases. RESULTS: CBF was significantly greater in the ET-1-injected eyes than in the control eyes 40-130 min after injection of ET-1 (P < 0.05). The maximal CBF ratio in the ET-1-injected eyes was 128 +/- 7.4% at 40 min. CBF decreased to the pre-injection level at 140 min after the injection of ET-1. There was no significant change in blood pressure, pulse rate and blood gases throughout this experiment, and there was no significant difference in IOP between ET-1-injected eyes and control eyes. CONCLUSION: It seems likely that the increase in CBF resulted from some local mechanisms of control that compensated for the decrease in RBF induced by intravitreal injection of ET-1. This model may be useful for investigation of the regulatory system of intraocular circulation, including endothelin receptors.

Frequency of glutathione-S-transferase 1 gene deletion and its possible correlation with cataract formation.

This experiment was conducted to investigate the possible association between an increased frequency of glutathione-S-transferase (GST)1 gene deletion and the presence of cataracts in elderly patients. Genomic DNA was isolated from blood samples obtained from 138 elderly patients who had undergone cataract surgery, and from 62 random blood donors. All subjects lived in the same geographic area (Ibaraki Prefecture, Japan). The DNA sequences among three different exon ranges (exons 3-5, exons 4-5 and exons 5-6) of the GST1 gene were amplified by the polymerase chain reaction (PCR) technique to determine if GST1 gene deletion occurred. Cataract patients had a significantly higher frequency of GST1 gene deletion than random controls did (P < 0.001, odds ratio = 2.91, 1.56-5.44; 95% of confidence interval). Mean age of cataract patients lacking GST1 gene was significantly younger (n = 101, mean age = 70.4, s.d. = 10.2) than that of patients possessing the GST1 gene (n = 37, mean age = 75.0, s.d. = 8.7) (P < 0.02). These results show that the deletion of the GST1 gene may be one of determinants of genetic susceptibility to cataractgenic agents.

Optimum form of posterior chamber intraocular lenses to minimize aberrational astigmatism.

To optically determine the optimum form for a posterior chamber intraocular lens (PC IOL), we calculated the aberrational astigmatism induced by tilt and decentration of the PCIOL using an exact raytracing. First, the position and the radii of curvatures of the IOL were determined to make an emmetropic eye model using a paraxial raytracing. Next, the chief rays originating from the fovea centralis were traced backward through the tilted and/or decentrated PC IOL, the center of the pupil and the cornea, using trigonometric raytracing. Finally, the maximum and minimum aberrational astigmatism were calculated based on the Coddington's Equations for the sagittal and the tangential foci of the ray. All the refractive parameters in Gullstrand's No. 1 schematic eye were adopted. The effect of varying anterior corneal asphericity on the results was also examined. Four forms of polymethylmethacrylate PC IOLs (refractive index: 1.491) were analyzed; a plano-convex IOL with the curved surface facing the cornea, and three bi-convex forms with the ratio of anterior-to-posterior radii of curvatures of 1:4, 1:2 and 1:1, respectively. The 1:4 bi-convex form showed the lowest values for the maximum aberrational astigmatism calculated at every combination of tilt and decentration except 0 degrees tilt and/or 0 mm decentration. The aberrational astigmatism with the 1:4 bi-convex form of PC IOL did not exceed 1.0 D at the maximum tilt and decentration. The variation of anterior corneal asphericity did not influence the results. We conclude that the 1:4 bi-convex form of PC IOL minimizes the postoperative astigmatism induced by tilt and/or decentration of the lens.

[Acanthamoeba isolation from contact lens solution of contact lens wearers without keratitis].

Acanthamoeba keratitis seems to be associated with wearing contact lenses. As controls, we surveyed contact lens wearers without keratitis. Contact lens solutions of 93 persons were examined in order to identify risk factors for contamination by Acanthamoeba. Therefore, the contact lens disinfection system and storage schedules were studied in each case. Acanthamoeba organisms were isolated from 4 specimens (4.3%). The incidence of Acanthamoeba was higher in specimens of soft contact lens solution than in those of hard contact lens solution, and all the Acanthamoeba positive cases had been using tap water.

A new model of transient complete obstruction of retinal vessels induced by endothelin-1 injection into the posterior vitreous body in rabbits.

Endothelin-1 (ET-1) is a potent vasoconstrictor peptide produced by vascular endothelial cells. In order to investigate the effects of ET-1 on retinal vessels, ET-1 (1-1000 pmol) was injected into the posterior vitreous body in rabbits. A high dose of ET-1 induced transient complete obstruction of the retinal vessels. In this experimental model of transient complete obstruction of the retinal vessels, the effects of ET-1 on retinal function were further analyzed by means of electroretinograms. The scotopic a-wave was not affected, but the amplitude of the scotopic b-wave was significantly elevated. The amplitude of oscillatory potentials was significantly reduced. These phenomena suggested that retinal ischemia without choroidal ischemia was brought about due to severe vasoconstriction of the retinal arteries. These findings indicate that intravitreal injection of ET-1 causes a transient cessation of blood supply from retinal vessels and that oscillatory potentials in electroretinograms appear to be sensitive for detecting changes of retinal circulation. This new model of transient complete obstruction of retinal vessels might be useful for studying the pathophysiology of severe retinal ischemia.

[Endothelin-1-induced vasoconstriction in retinal blood vessels in the rabbit].

Endothelin-1 (ET-1) is an endothelium-derived potent vasoconstrictor peptide which is synthesized in retinal vessels. In order to investigate its effect on retinal blood vessels, we injected ET-1 intravitreally (10(-12), 3 x 10(-12), 10(-11), 3 x 10(-11), 10(-10) mole) and examined the change in diameters of retinal arteries in rabbits by fundus photography. After injection of a high dose (10(-10) mole) of ET-1, fluorescein angiography was used to investigate whether complete occlusions of retinal blood vessels occurred or not. 3 x 10(-12) mole ET-1 induced contraction of retinal arteries and the contraction of those induced by 10(-10) mole ET-1 showed complete occlusion. A medium dose (10(-11) mole) of ET-1 caused repeated vasospasms and segmental vasoconstrictions of retinal blood vessels. Fluorescein angiograms revealed that retinal blood flow was interrupted transiently by injection of a high dose (10(-10) mole) of ET-1. These findings suggested that intravitreal injection of ET-1 could provide a new model of experimental transient occlusion of retinal vessels in rabbits.

Analysis of the components of electrically evoked response using a monopolar recording technique.

PURPOSE: To assess the significance of the components of the electrically evoked response. METHODS: Twenty-three normal subjects, six patients with macular holes, and three patients with retinal artery occlusion were tested. The electrically evoked responses were recorded from an occipital electrode by applying an electric current of 0.3 mA-2.0 mA with a duration of 5 msec and a frequency of 1.85 Hz. RESULTS: In the normal subjects, three rhythmic waves (N1P1, N2P2, N3P3) were frequently found with fewer individual differences in their latencies than in flash visual evoked potentials. It was found that the amplitude of N1P1 became larger in proportion to the stimulus strength, and that the amplitudes of N2P2 and N3P3 reached their ceiling peaks. The amplitudes of N2P2 were significantly reduced in the affected eyes of patients with a unilateral complete break of the macula and branch retinal artery occlusion including the macular area. Conversely, in one patient with central retinal artery occlusion, whose visual acuity was good because the cilioretinal artery was patent, the amplitude of N1P1 was significantly reduced in the affected eye. CONCLUSIONS: N2P2 in electrically evoked response might originate mainly in the macular area. The analysis of N2P2 may be useful for further clinical applications of electrically evoked response.