Temporary amniotic membrane patching for acute chemical burns.

PURPOSE: To describe the surgical technique, and its usefulness, of temporary amniotic membrane patching (AMP) in the acute phase of ocular chemical injury. METHODS: Temporary AMP with modification in suture placement was performed on five eyes of five consecutive patients inflicted with acute chemical injury having a greater than grade II injury by the Roper-Hall classification. RESULTS: All patients reported herein presented with a large epithelial defect on the cornea and conjunctiva. Case 3 was classified as grade III while the other four cases were classified as grade II. The causative chemical agents were anhydrous acetic acid in Case 1, calcium oxide in Case 2, sodium hydroxide in Case 3, sodium silicate in Case 4, and sulphuric acid in Case 5. All cases experienced rapid relief of pain after AMP. Epithelialization of the cornea with improvement of visual acuity was observed in all cases when the amniotic membrane was removed within 2 weeks after surgery. During the mean follow-up of 19.6 months, the ocular surface remained stable and no cicatricial complications were noted. CONCLUSIONS: These results suggest that immediate AMP is quite useful for managing moderately severe acute ocular chemical injury by facilitating rapid epithelialization and pain relief, and securing ocular surface integrity.

Multi-layer amniotic membrane graft for pterygium in a patient with xeroderma pigmentosum.

BACKGROUND: Xeroderma pigmentosum (XP) is a rare, autosomal recessive, premalignant condition of the skin, and is reported to be associated with ocular surface disorders such as conjunctival malignancy and pterygium. Herein, we report a case of successful management of pterygium with multi-layer amniotic membrane graft (AMG) in a young XP patient. CASE: An 11-year-old Japanese girl, who had been diagnosed as having XP, was referred to us for treatment of her bilateral pterygium. Surgical intervention was attempted for a temporal, presumably fast-growing pterygium in her left eye. Multi-layer amniotic membrane grafting was performed. OBSERVATIONS: The surgery-induced pain and irritation disappeared within a day. The limbal conjunctival autograft survived on the AMG and re-epithelialization over the AMG was completed in 2 weeks. Best corrected visual acuity improved from 20/32 to 20/16 one month postoperatively. During the 1-year follow-up period, no recurrence was noted. CONCLUSIONS: The present case exemplifies that AMG as an adjunct to primary pterygium resection is effective even in a young patient with XP. In addition, multi-layer AMG, which we first demonstrated in this report, seems to be useful for protecting bare sclera and extraocular muscles from mechanical injury.

Two types of K+ currents modulated by arachidonic acid in bovine corneal epithelial cells.

PURPOSE: Fenamate sensitivity of the large-conductance K+ current in the corneal epithelium suggests that K+ transport could be modulated by arachidonic acid (AA) and/or its metabolites, which also regulate corneal epithelial migration. The main purpose of this study was to investigate AA-induced modulation of K+ currents expressed in the bovine corneal epithelium. METHODS: Freshly isolated bovine corneal epithelial cells were perfused with Ringer solution. Whole-cell currents were recorded by using either the conventional whole-cell-patch or the perforated-patch configuration. RESULTS: Two distinct types of K+ currents dominated the whole-cell current. The first was a voltage-gated K+ current that was inactivated completely by membrane depolarization. The inactivating voltage-gated K+ current was largest in presumptive basal cells. The second was a noisy, sustained K+ current that was never inactivated and seemed to be a counterpart of the large-conductance K+ current reported in the rabbit corneal epithelium. External application of AA (5-20 microm) inhibited the inactivating voltage-gated K+ current and augmented the noisy, sustained K+ current. Identical dual modulation was induced by other fatty acids (e.g., palmitoleic acid) that are not substrates for enzymes in the AA cascade. CONCLUSIONS: An inactivating voltage-gated K+ channel was identified for the first time in the corneal epithelium. AA and some fatty acids may directly activate the large-conductance K+ channel to augment its housekeeping functions in corneal epithelial cells.

Scanning Laser Ophthalmoscopic Microperimetry on Idiopathic Epiretinal Membrane and Vitreomacular Traction Syndrome.

Purpose: To evaluate visual function before and after vitrectomy for idiopathic epiretinal membrane and vitreomacular traction syndrome.Methods: Microperimetry with scanning laser ophthalmoscope (SLO) was performed in 4 eyes with idiopathic epiretinal membrane (IERM) and 2 eyes with vitreomacular traction syndrome (VMTS), before and at least 3 months after vitrectomy.Results: Five eyes had preoperative scotoma, which disappeared postoperatively. In 1 eye, absolute and relative scotomas developed postoperatively over the areas where the retinal surface was vigorously scratched to peel off an overlying membrane, and in another eye new scatomas appeared over a newly formed epiretinal membrane 8 months after vitrectomy.Conclusions: To our knowledge, the present report is the first to describe preoperative absolute scotoma in IERM, VMTS and also iatrogenic absolute scotoma due to retinal scratching. SLO microperimetry is useful for evaluation of visual function before and after vitrectomy for epiretinal membrane and vitreomacular traction syndrome.

Identification of a novel fluorophore, xanthurenic acid 8- O -beta-D-glucoside in human brunescent cataract.

We have identified the chemical structure of a novel protein-unbound fluorescent glucoside (Fl-Glc), found to be far more abundant in the human brunescent cataractous lens nuclei than in non-brunescent ones. Our earlier experiments showed that long-term incubation of the protein-free filtrate of non-brunescent cataractous nuclei generated increasing amounts of a particular yet to be characterized fluorophore (Fl-X). High performance liquid chromatography (HPLC) analyses revealed Fl-X and Fl-Glc to be identical. HPLC-electrospray ionization-mass spectrometry (HPLC-ESI-MS) disclosed the molecular weights (MW) of Fl-X and its beta-glucosidase-digest (Fl-X-aglycon) to be 367 and 205, respectively. Fl-X-aglycon and authentic xanthurenic acid (MW = 205) not only eluted at exactly the same retention time on HPLC but also revealed their protonated ions at the same m/z of 206.1 by positive ion analysis on HPLC-ESI-MS. These results suggest that Fl-X ( = Fl-Glc) is a beta-glucoside of xanthurenic acid. Fl-Glc was finally identified as xanthurenic acid 8- O -beta- D -glucoside because the retention times of both completely agreed with three kinds of HPLC conditions.

Identification of a Free Non-tryptophan Fluorophore in Water-soluble Fraction of Human Brunescent Cataractous Lens Nucleus.

Purpose: We previously reported that a unique free fluorophore (Fl-Glc), presumably a beta-glucoside, is particularly abundant in human brunescent cataractous lens nuclei. Our preliminary experiments indicated that incubation of low-molecular weight (MW) fraction of non-brunescent lens nuclei causes an increase in a particular fluorophore (Fl-X). This study was undertaken to compare the Fl-Glc with the Fl-X and subsequently to identify the Fl-X.Methods: Experiment (1) The purified Fl-X and its beta-glucosidase digest (aglycon) were compared with the Fl-Glc and its aglycon, respectively, by high-performance liquid chromatography (HPLC). Experiment (2) i) The Fl-X and its aglycon were analysed by liquid chromatography/mass spectrometry (LC/MS). ii) Authentic xanthurenic acid was analysed by HPLC and LC/MS.Results: Experiment (1) The retention times of the Fl-X and the Fl-Glc exactly coincided. The fluorescence peaks of both disappeared after beta-glucosidase treatment. Experiment (2) i) LC/MS results suggested that the MWs of the Fl-X and its aglycon were 367 and 205, respectively. ii) HPLC and LC/MS results for xanthurenic acid (MW = 205) were exactly the same as those for the aglycon of the Fl-X.Conclusions: The Fl-Glc and the Fl-X are identifical, and the Fl-X (= Fl-Glc) is a glucoside of xanthurenic acid.

[Scanning laser ophthalmoscopic microperimetry on idiopathic epiretinal membrane and vitreomacular traction syndrome].

PURPOSE: To evaluate visual function before and after vitrectomy for idiopathic epiretinal membrane and vitreomacular traction syndrome. METHODS: Microperimetry with scanning laser ophthalmoscope (SLO) was performed in 4 eyes with idiopathic epiretinal membrane (IERM) and 2 eyes with vitreomacular traction syndrome (VMTS), before and at least 3 months after vitrectomy. RESULTS: Five eyes had preoperative scotoma, which disappeared postoperatively. In 1 eye, absolute and relative scotomas developed postoperatively over the areas where the retinal surface was vigorously scratched to peel off an overlying membrane, and in another eye new scotomas appeared over a newly formed epiretinal membrane 8 months after vitrectomy. CONCLUSIONS: To our knowledge, the present report is the first to describe preoperative absolute scotoma in IERM, VMTS and also iatrogenic absolute scotoma due to retinal scratching. SLO microperimetry is useful for evaluation of visual function before and after vitrectomy for epiretinal membrane and vitreomacular traction syndrome.

Remission of antiepiligrin (laminin-5) cicatricial pemphigoid after excision of gastric carcinoma.

PURPOSE: To describe a case of antiepiligrin cicatricial pemphigoid with unusual ocular manifestations and its remission after surgical removal of gastric carcinoma. METHODS: We describe a 61-year-old Japanese man with antiepiligrin cicatricial pemphigoid. RESULTS: He presented with conjunctival injection and discharge preceded by a 6-month period of erosive lesions in the oral mucosa and the truncal skin. An advanced gastric carcinoma was found and his serum immunoprecipitated laminin-5. Despite topical treatment with betamethasone, ofloxacin, and artificial tear solutions, serious symblepharon along the Meibomian line developed with little shortening of the inferior conjunctival sac. Following radical gastrectomy, the ocular and cutaneous lesions turned completely quiet. CONCLUSION: The present case differed from past cases by lacking inferior conjunctival sac shortening and by showing erosive lesions solely at the mucocutaneous junctions. The ocular involvement in this case correlated very well with the severity of gastric carcinoma.

Comparison of non-tryptophan fluorophores in protein-free extract of brunescent and non-brunescent human cataract.

PURPOSE: To investigate whether any of the fluorophores in the human lens nuclei might be responsible for human brunescent cataract formation. METHODS: Human lens nuclei (non-brunescent, from 13; brunescent, from 8) were obtained after extracapsular cataract extraction in nondiabetic patients. Protein-free extract, prepared by filtrating the water-soluble fraction of each nucleus through a centrifugal ultrafilter (molecular weight < 5,000), was analytically separated by high-performance liquid chromatography. RESULTS: No significant differences between non-brunescent and brunescent nuclei were observed in the concentrations (mean +/- SD) of 3-hydroxykynurenine O-beta-glucoside (0.67 +/- 0.38 vs. 0.85 +/- 0. 62 micromol/g wet weight), 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid O-glucoside (4.1 x 10(5) +/- 2.9 x 10(5) vs. 6.3 x 10(5) +/- 5. 0 x 10(5) area unit/g wet weight), and kynurenine (0.016 +/- 0.011 vs. 0.029 +/- 0.021 micromol/g wet weight). A novel fluorophore that has not been identified so far was significantly present more in brunescent than in non-brunescent nuclei (brunescent: 1.5 x 10(5) +/- 1.0 x 10(5) vs. non-brunescent: 2.6 x 10(3) +/- 6.3 x 10(3) area unit/g wet weight, P <.01). Digestion of the protein-free extract with beta-glucosidase eliminated the peak corresponding to the novel unidentified fluorophore. CONCLUSION: The present results imply a novel protein-unbound fluorophore, presumably a beta-glucoside, might possibly be involved in brunescent cataract formation.

[Identification of a free non-tryptophan fluorophore in water-soluble fraction of human brunescent cataractous lens nucleus].

PURPOSE: We previously reported that a unique free fluorophore (Fl-Glc), presumably a beta-glucoside, is particularly abundant in human brunescent cataractous lens nuclei. Our preliminary experiments indicated that incubation of low-molecular weight (MW) fraction of non-brunescent lens nuclei causes an increase in a particular fluorophore (Fl-X). This study was undertaken to compare the Fl-Glc with the Fl-X and subsequently to identify the Fl-X. METHODS: Experiment 1. The purified Fl-X and its beta-glucosidase digest (aglycon) were compared with the Fl-Glc and its aglycon, respectively, by high-performance liquid chromatography (HPLC). Experiment 2. i) The Fl-X and its aglycon were analysed by liquid chromatography/mass spectrometry (LC/MS). ii) Authentic xanthurenic acid was analysed by HPLC and LC/MS. RESULTS: Experiment 1. The retention times of the Fl-X and the Fl-Glc exactly coincided. The fluorescence peaks of both disappeared after beta-glucosidase treatment. Experiment 2. i) LC/MS results suggested that the MWs of the Fl-X and its aglycon were 367 and 205, respectively. ii) HPLC and LC/MS results for xanthurenic acid (MW = 205) were exactly the same as those for the aglycon of the Fl-X. CONCLUSIONS: The Fl-Glc and the Fl-X are identical, and the Fl-X (= Fl-Glc) is a glucoside of xanthurenic acid.

A retrospective study of ocular toxocariasis in Japan: correlation with antibody prevalence and ophthalmological findings of patients with uveitis.

To classify the clinical characteristic of ocular toxocariasis in Japan, the prevalence of antibodies to Toxocara antigens was examined in patients with uveitis of unknown aetiology. From 1982 to 1993, serum specimens of 383 cases and intraocular fluid samples of 22 cases were serologically screened for Toxocara infection with five immunodiagnostic tests. Fifty-five sera and 11 intravitreous fluid samples were estimated to have significantly high antibody levels against larval excretory-secretory (ES) antigens of T. canis. Eight cases were positive in both serum and vitreous fluid, and three were positive only in the vitreous fluid. Among the 58 antibody positive samples, 20 cases were omitted due to a lack of detailed description of ocular findings. The remaining 38 cases are described in this study. Of these 38 cases, 34 (89%) were older than 20 years of age. Ocular lesions were located in the posterior fundus in 11 cases, in the peripheral fundus in 18 cases, and in both areas in seven cases. Of the eight cases in which papillary oedema or redness was observed, chorioretinal lesions were also present in seven of them. Tractional retinal detachment was present in five cases. These observations suggest that ocular toxocariasis in Japan has a different clinical profile compared with those in the other countries, and indicate a need for revised classification of ocular toxocariasis.

Upregulation of retinal vascular endothelial growth factor mRNAs in spontaneously diabetic rats without ophthalmoscopic retinopathy. A possible participation of advanced glycation end products in the development of the early phase of diabetic retinopathy.

Vascular endothelial growth factor (VEGF) has recently been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, its involvement in the development of the early phase of diabetic retinopathy is not fully understood. In this study we investigated the retinal VEGF mRNA level in spontaneously diabetic Otsuka Long-Evans Tokushima fatty (OLETF) rats, a model of non-insulin-dependent diabetes, without overt retinopathy, using quantitative reverse-transcription polymerase chain reaction. The retinal VEGF mRNA level was 2.2 times higher (p < 0.0005) in OLETF rats than in control rats at the age of 60 weeks. Moreover, their retinal mRNA level was positively correlated with serum concentration of advanced glycation end products (AGEs) but not to serum glucose concentration. Furthermore, the peak latency of the oscillatory potentials in the electroretinogram, one of the most sensitive markers for the early phase of diabetic retinopathy, was significantly prolonged in OLETF rats (p < 0.05), being also correlated with the serum AGE concentration. The results thus suggest that AGEs, which are formed acceleratedly in diabetic conditions, are involved in the development of the early phase of diabetic retinopathy probably through the induction of retinal VEGF mRNAs.

Electrical responses from diabetic retina.

Diabetic retinopathy has long been considered to be a retinal manifestation of systemic diabetic angiopathy. Indeed, it is therapeutically true. However, the prolongation of OP peak latency in diabetic eyes without any angiographic evidence of angiopathy leads us to presume that certain neuronal disorders occur early in diabetic eyes. Even though we cannot neglect the possibility that the prolongation of the OP peak latency may derive from undetectable retinal hypoperfusion, it is still far from conventional diabetic angiopathy. Rather, the status should be properly termed "intraretinal diabetic neuropathy" in that the neurones are the disturbed cells to cause visual dysfunction. Thereafter, the OP amplitude diminishes as retinopathy advances, probably depending on the degree of retinal circulatory disturbance. Marked diminution of the OP amplitude predicts rapid progression and poor prognosis of retinopathy. Diabetic retinal pigment epitheliopathy as manifested by one of our non-photic EOG responses is another kind of early ocular involvement of diabetes. Because its mechanisms are not yet known, so far we have not succeeded in correlating it to any kind of subjective visual index. Routine fundus inspection or fluorescent fundus angiography is incapable of detecting the compromised neural retina and/or retinal pigment epithelial integrity and thus the electrophysiology of vision has the edge in ophthalmology.

Retinal pigment epithelial origin of bicarbonate response.

The mechanisms of the bicarbonate-induced decrease in the ocular standing potential (bicarbonate response) were investigated in the cat. An intravenous infusion of 1.4% sodium bicarbonate solution caused a decrease in the standing potential. A high-bicarbonate solution decreased or increased the potential across the retinal pigment epithelium-choroid tissue of the excised cat eye when applied basally or apically, respectively, but did not affect the potential across the anterior portion of the excised eye or across the isolated neural retina. A high-bicarbonate solution principally depolarized the apical membrane of the retinal pigment epithelium (RPE) when applied basally, and hyperpolarized it when applied apically. These results suggest that the bicarbonate response in the cat is primarily due to the effects of an increase in bicarbonate concentration on the basal membrane of the RPE.

Differentiation of neovascular maculopathies by nonphotic electrooculogram responses.

Four kinds of electrooculographic (EOG) responses for the retinal pigment epithelium (RPE) were studied to assess RPE integrity in neovascular macular diseases. In 20 eyes with age-related macular degeneration (ARMD), 13 fellow eyes of ARMD patients, eight eyes with central exudative chorioretinopathy (CECR), and seven unaffected eyes of CECR patients, we evaluated the light peak/dark trough ratio, and the hyperosmolarity, Diamox, and bicarbonate responses. We found no abnormalities in any ARMD eyes or fellow eyes of ARMD patients. In both the CECR eyes and unaffected eyes of CECR patients, however, the Diamox response was subnormal whereas the other three EOG responses remained normal. There was no significant difference in Diamox response between the CECR eyes and unaffected eyes of CECR patients. The selective deterioration of the Diamox response in CECR, but not in ARMD, can serve as a new index for differentiating between these similar conditions.

Neural retinal contribution to the slow negative potential of the canine electroretinogram.

To establish the normal waveform of the electrical responses from canine eyes, electroretinograms and the light peak were recorded in hybrid and beagle dogs under general anesthesia and artificial ventilation. The neural retinal and retinal pigment epithelial components were pharmacologically isolated by intravitreal glutamate injection and systemic sodium iodate administration, respectively. The a- and b-waves elicited by either flash or rectangular stimuli, the oscillatory potentials elicited by flash stimuli and the light peak elicited either by a single maintained illumination or by repetitive stimuli were almost identical with those of other vertebrates thus far studied. In contrast, in response to rectangular (several-second duration) stimuli, the c-wave was usually absent and was replaced by a slow cornea-negative potential that had a time course similar to that of the c-wave in other species. This slow negative potential was elicited at such low stimulus intensities that the a-wave was absent, was deepened by an intravenous administration of sodium iodate, was not affected by an intravitreal injection of sodium glutamate and was shallowed during the light peak. These results suggest that the slow negative potential that replaces the c-wave consists of large slow PIII, small retinal pigment epithelial c-wave and negligible contribution from the late photoreceptor potential and the dc component of the PII.

Normal S cone electroretinogram b-wave in Oguchi's disease.

AIM: The short wavelength sensitive (S) cone electroretinograms (ERGs) were examined in two patients with Oguchi's disease to study S cone function. METHODS: Ganzfeld colour flashes under bright white background illumination were used to elicit S cone, and mixed long (L) and middle (M) wavelength sensitive cone ERGs. RESULTS: The S cone ERG b-wave was normal with short wavelength stimuli with normal L and M cone responses in both patients. CONCLUSIONS: These ERG results indicate that the S cone system as well as the L and M cone system is not defective in Oguchi's disease, while the S cone responses are not detectable in the complete type of congenital stationary night blindness as reported previously. The results imply that there is a significant difference in the way the S cone system is affected in different forms of stationary night blindness, and that the S cone and rod do not share the same arrestin system.