Large-scale flooding analysis in the suburbs of Tokyo Metropolis caused by levee breach of the Tone River using a 2D hydrodynamic model.

In order to assess the effects of climate change on flood disasters in urban areas, we applied a two dimensional finite element hydrodynamic model (2D-FEM) to simulate flood processes for the case analysis of levee breach caused by Kathleen Typhoon on 16 September 1947 in Kurihashi reach of Tone River, upstream of Tokyo area. The purpose is to use the model to simulate flood inundation processes under the present topography and land-use conditions with impending extreme flood scenarios due to climate change for mega-urban areas like Tokyo. Simulation used 100 m resolution topographic data (in PWRI), which was derived from original LiDAR (Light Detection and Ranging) data, and levee breach hydrographic data in 1947. In this paper, we will describe the application of the model with calibration approach and techniques when applying for such fine spatial resolution in urban environments. The fine unstructured triangular FEM mesh of the model appeared to be the most capable of introducing of constructions like roads/levees in simulations. Model results can be used to generate flood mapping, subsequently uploaded to Google Earth interface, making the modeling and presentation process much comprehensible to the general public.

Primary amyloidosis with familial vitreous opacities: an unusual case and family.

Peripheral neuropathy was not found even six to ten years after the onset of visual symptoms in a family with primary amyloidosis, except in the propositus at the terminal stage. The propositus had mainly ocular and CNS involvement. An ocular manifestation, the vitreous opacity, was the only involvement in the family members, in spite of the long clinical course. This family may have a different type of familial primary amyloidosis from that previously reported.

Retinal functional change caused by adenoviral vector-mediated transfection of LacZ gene.

We examined the effect of insertion of an exogenous gene on retinal function to assess the rationale of adenoviral vector-mediated gene transfer for future gene therapy. An adenoviral vector expressing bacterial LacZ (AdCALacZ) was injected into the eyes of adult rats either intravitreally (group A) or subretinally (group B), and the gene expression and retinal function were thus examined at different time points after gene transfer for 3 weeks. X-Gal histostaining showed that neural retinal cells were transfected in group A and that retinal pigment epithelial cells were transfected in group B. The gene transfer was more efficient in group B (54.4% of the fixed retinal area was stained) than in group A (10.4%). The electroretinogram (ERG) revealed retinal dysfunction in the AdCALacZ-transfected rats even at the stage in which the histological damage was not apparent by electron microscopy and immunohistochemical studies for cytokeratin, S-100 protein, and glial fibrillary acidic protein. The ERG change was correlated with the intensity of inflammation, and retinal function recovered to the original level by 3 weeks, along with a diminution of inflammation. Functional changes were more evident in eyes treated with AdCALacZ than in those infected with adenoviral vector with no exogenous gene; however, no histological difference was observed between these groups, indicating that the insertion of exogenous gene itself affects retinal function. The results showed that different kinds of retinal cells could be gene-transferred by an adenoviral vector, depending on the application method. The retinal dysfunction caused by each adenoviral transfection method was caused by inflammation and the insertion of exogenous gene, and this retinal dysfunction was recoverable. In future gene therapy, special attention should be given to the method of exogenous gene insertion in the retina.

Target gene transfer of tissue plasminogen activator to cornea by electric pulse inhibits intracameral fibrin formation and corneal cloudiness.

Intracameral fibrin formation, a complication of ocular inflammation and intraocular operations, sometimes results in glaucoma and/or corneal damage leading to permanent visual loss. We transferred a therapeutic gene to the corneal endothelium in order to use it as a therapeutic organ. A plasmid encoding tissue plasminogen activator (tPA) was injected into the anterior chamber of rats and electric pulses (EPs) were given subsequently, which transferred a plasmid gene to a highly selected area of corneal endothelium with no inflammation. The biologically active tPA was clearly present for 4 days after treatment. Fibrin formation induced by YAG laser-generated bleeding in the anterior chamber decreased significantly more in treated eyes than in control eyes. Corneal opacity was significantly lower in treated eyes than in control eyes and histological damage was not apparent in the treated eyes. This genetic modification allows us to use the corneal endothelium to treat various ocular diseases and could be a new and effective type of pharmacologic gene therapy.

High prevalence of antineutrophil cytoplasmic antibody positivity in childhood onset Graves' disease treated with propylthiouracil.

Propylthiouracil (PTU)-induced antineutrophil cytoplasmic antibody (ANCA)-related vasculitis and nephritis were recently reported in about 30 patients with hyperthyroidism. The objective of this study was to clarify the prevalence of ANCA and the relationship between ANCA and thyroid antibodies in children with Graves' disease. Titers of myeloperoxidase (MPO)-ANCA in sera of 51 patients with childhood onset Graves' disease (16 before treatment, 25 and 10 treated with PTU and methimazole, respectively) were measured by enzyme-linked immunosolvent assay. Antithyroglobulin antibodies (TGAbs) and antithyroperoxidase antibodies (TPOAbs) were also measured by RIA in 25 PTU-treated patients. No patients had clinical manifestations of vasculitis and nephritis. MPO-ANCA was positive in 6.7% of patients before treatment and in 64.0% of those treated with PTU and in none of those treated with methimazole. MPO-ANCA had a significantly positive correlation with TGAbs (P < 0.05) and no significant correlation with TPOAbs. These findings show the high prevalence of the MPO-ANCA positivity in PTU-treated childhood onset Graves' disease, suggesting that PTU may not be preferred as the first line for the treatment of children with Graves' disease. The significant correlation between MPO-ANCA and TGAbs indicates that the severity of Graves' disease may be a factor responsible for the MPO-ANCA positivity. The cross-reactivity between MPO-ANCA and TPOAbs may not play a role in the high prevalence of MPO-ANCA in the patients exposed to PTU.

High prevalence of T354P sodium/iodide symporter gene mutation in Japanese patients with iodide transport defect who have heterogeneous clinical pictures.

A missense and loss of function mutation of the Na+/I- symporter (NIS) gene, T354P [Thr354-->Pro (ACA-->CCA)], was found in the homozygous state in two unrelated Japanese patients with iodide transport defect. In this study we have identified the homozygous T354P NIS germline mutation in seven Japanese patients, including one previously reported, from five unrelated families. No other nucleotide changes were found in the coding regions and the exon-intron boundaries of the NIS gene in these seven patients. These results suggest a common prevalence of the T354P mutation in Japanese patients. Although these seven patients have the identical NIS mutation, T354P, marked heterogeneity in clinical pictures, especially concerning goiter and hypothyroidism, were noted among them. Therefore, another factor(s), but not the nature of the NIS mutation, may account for the clinical heterogeneity among patients with the iodide transport defect. We have previously reported that the NIS messenger ribonucleic acid was markedly increased in the thyroid of a patient with the homozygous T354P mutation. In this study we demonstrated that the NIS proteins in the patients' thyroids were significantly increased (approximately 10-fold) by Western blot analysis of integral membrane proteins using an antibody against the C-terminal peptide of the human NIS. Furthermore, we showed by immunohistochemical staining that the T354P mutant NIS proteins were overexpressed in the basal and lateral plasma membranes of patients' thyrocytes.

The role of NF-kappaB in retinal neovascularization in the rat. Possible involvement of cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin-8 family.

Hypoxia precedes neovascularization in many retinal diseases that can lead to irreversible vision loss. The transcription factor NF-kappaB is activated by hypoxia and regulates the expression of many genes, including angiogenic factors. The relation between the NF-kappaB activation and the cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin-8 (IL-8) family, was investigated by immunohistochemistry in a rat model of proliferative retinopathy presumably caused by relative hypoxia. Activated NF-kappaB and CINC immunoreactivity was detected in retinal glial cells in the nonperfused retina and in neovascular cells. Activated NF-kappaB was detected before the CINC staining, and both of these events occurred before the development of neovascularization. The intensity of both activated NF-kappaB and CINC staining remained increased during the development of neovascularization and then declined as neovascularization regressed. In rat retinal glial cells in vitro, dexamethasone, an inhibitor of NF-kappaB activation, prevented the hypoxia-induced increase in the amount of CINC mRNA. Furthermore, CINC induced neovascularization in a rat corneal pocket model. These results suggest that hypoxia-induced activation of NF-kappaB results in CINC production and participates in the induction of retinal neovascularization.

Relation between superficial capillaries and foveal structures in the human retina.

When examining semithin Epon sections of human retinas, it became evident that superficial capillaries showed four different positions, according to the thickness of the ganglion cell layer. For the clear view of the distribution of the position of superficial capillaries, the intrafoveal region was subdivided into four zones, based on the thickness of the ganglion cell layer; the foveola and the A-, B-, and C-zone. The foveola has no ganglion cell layer, and the A-zone has a ganglion cell layer thinner than 15 microns. These regions lack superficial capillaries. In the B-zone, the ganglion cell layer is 15-45 microns thick, and here the superficial capillaries lie in the outer boundary of the ganglion cell layer. The C-zone and parafovea have a ganglion cell layer thicker than 45 microns, and superficial capillaries are present within the ganglion cell layer. The perifovea has a ganglion cell layer 15-45 microns thick. In the temporal perifovea, where the nerve fiber layer is not so distinct, superficial capillaries are located on the outer boundary of the ganglion cell layer. In the other portion of the perifovea, superficial capillaries lie in the inner boundary of the ganglion cell layer. Out of the perifovea, where the ganglion cell layer is thinner than 15 microns, most of superficial capillaries touch both boundaries of the ganglion cell layer. Major retinal vessels touch the ganglion cell layer and lie in the similar position to that of superficial capillaries.

Lipofuscin granules in human photoreceptor cells.

Twenty-four human retinas were structurally examined in order to study the degradative pathway in inner segment turnover. Lipofuscin granules were found in myoids of photoreceptor inner segments. Cone lipofuscin granules exhibited autofluorescence, in ultraviolet light. Both the cone lipofuscin granules (-1.6 micron) and the rod ones (-0.6 micron) were membrane-limited inclusions comprising different contents. Various vacuoles related to autophagy were found in the myoids of rods and cones. Acid phosphatase activity was demonstrated in lipofuscin granules of the rods and cones, as well as adjacent various vacuoles. A survey of the cone lipofuscin granules in the semithin Epon sections revealed that more than one-half of the eyes from humans over 30 years of age contained cone lipofuscin granules, whereas eyes from those under 30 years of age did not. These results strongly suggest that lipofuscin granules represent an accumulation of residual bodies of autophagy in the photoreceptor inner segments.

Differential effects of diltiazem and nitroglycerin on cytosolic Ca2+ concentration and on force in the bovine ophthalmic artery.

PURPOSE: To determine the mechanisms of inhibition by diltiazem (Dil) and nitroglycerin (NG) of the contraction induced by serotonin (5-HT) in the ophthalmic artery. METHODS: Using front-surface fluorometry of fura-2 and the medial strips of the bovine ophthalmic artery, [Ca2+]i and force were monitored simultaneously. Changes in the force at a constant [Ca2+]i were determined by use of receptor-coupled membrane permeabilization with alpha-toxin. RESULTS: In the presence of extracellular Ca2+, 5-HT (10(-5) M) induced an initial transient and subsequently lower steady state elevation of [Ca2+]i. The transient elevation of [Ca2+]i was dependent on both intracellular and extracellular [Ca2+]i, whereas the steady state elevation was dependent on only extracellular Ca2+. For a given level of elevation of [Ca2+]i, 5-HT produced a greater force than the depolarization with high external K+ (118 mM) solution. In the permeabilized ophthalmic artery smooth muscle, 5-HT enhanced the contractile response to constant cytosolic Ca2+ (pCa 6.5) in the presence of guanosine triphosphate (GTP, 10 microM), but not in its absence. Therefore, 5-HT induces [Ca2+]i elevation, depending on both extracellular (Ca2+ influx) and intracellular Ca2+ (Ca2+ release), and it potentiates the Ca2+ sensitivity of the contractile apparatus through the activation of G-proteins. 5-HT-induced release of Ca2+ from the store was inhibited by NG, but not by Dil, in a concentration-dependent manner. However, neither NG nor Dil inhibited caffeine (20 mM)-induced release of Ca2+ from the store. Dil (10 microM) and NG (10 microM) inhibited in a concentration-dependent manner the steady state elevations of [Ca2+]i (Ca2+ influx) and force induced by 5-HT (10 microM) in the presence of extracellular Ca2+. Dil equally inhibited the steady state elevations of [Ca2+]i and force induced by 5-HT, whereas NG inhibited the force to a greater extent than expected from the reduction in [Ca2+]i. In the permeabilized ophthalmic artery smooth muscle, NG (10 microM), but not Dil (10 microM), decreased the force development induced by GTP (10 microM) and 5-HT (10 microM) at constant [Ca2+]i (pCa 6.5). These results indicate that NG, but not Dil, decreases the Ca2+ sensitivity of contractile apparatus. CONCLUSIONS: The authors found that 5-HT contracts the ophthalmic artery smooth muscle by the elevation of [Ca2+]i mediated by the release of intracellular Ca2+ and the influx of extracellular Ca2+, as well as by an increase in the Ca2+ sensitivity of the contractile apparatus through the activation of G-proteins, and that Dil relaxes 5-HT-mediated contraction of ophthalmic artery primarily by inhibiting the Ca2+ influx and, hence, by decreasing [Ca2+]i without having any effect on the Ca2+ sensitivity of the contractile apparatus. Nitroglycerin relaxes the ophthalmic artery not only by decreasing [Ca2+]i (inhibition of both the Ca2+ release and Ca2+ influx) but also by decreasing the Ca2+ sensitivity of the contractile apparatus.

Role of NF-kappaB-mediated interleukin-8 expression in intraocular neovascularization.

PURPOSE: To investigate the role of interleukin (IL)-8 in intraocular neovascularization and the mechanism of its production. METHODS: Interleukin-8 was measured with enzyme-linked immunosorbent assays in vitreous and aqueous fluid obtained from patients with neovascular diseases. Localization of IL-8 was examined by immunohistochemistry. An in vitro angiogenesis assay was performed on collagen gels, by using bovine aortic endothelial cells to determine the effect of the vitreous fluid. In bovine retinal glial cells under hypoxia, NF-kappaB activation was evaluated by immunoblot analysis and by electrophoretic mobility shift assay, and IL-8 and vascular endothelial growth factor (VEGF) mRNA expression was determined by semiquantitative reverse transcription-polymerase chain reaction. RESULTS: The concentration of IL-8 in vitreous fluid of patients with retinal neovascularization was significantly higher than that of patients without neovascular disease. Interleukin-8 immunostaining was detected in vascular endothelial cells and glial cells in the retinas with neovascularization. Vitreous fluid with high concentrations of IL-8 induced tubular morphogenesis in endothelial cells, and this effect was inhibited to a similar extent by neutralizing antibodies to IL-8 or to VEGF. In glial cells, in vitro, hypoxia induced NF-kappaB activation and increased IL-8 and VEGF mRNA. Furthermore, pyrrolidine dithiocarbamate, a specific inhibitor of NF-kappaB activation, prevented the induction of the IL-8 gene, but not that of the VEGF gene. CONCLUSIONS: These results suggest that IL-8 induced by hypoxia and mediated by NF-kappaB may contribute to the pathogenesis of intraocular neovascularization.

Suppression of retinal neovascularization by the NF-kappaB inhibitor pyrrolidine dithiocarbamate in mice.

PURPOSE: To evaluate the effect of pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappaB (NF-kappaB), on retinal neovascularization in a murine model of ischemic retinopathy. METHODS: One-week-old C57BL/6N mice were exposed to 75%+/-2% oxygen for 5 days and then were returned to room air to induce retinal neovascularization. After the return to room air, the left and right eyes were injected intravitreally with PDTC or a vehicle, respectively. Retinal neovascularization was examined by injecting fluorescein dextran and angiography after 5 days in room air and was quantitated histologically with a masked protocol. The effects of PDTC on NF-kappaB activation were evaluated by immunohistochemistry. To examine the toxicity of PDTC, the histologic change in the retina was examined by light and electron microscopy. RESULTS: Retinal neovascularization in the eye injected with PDTC by intravitreal methods was reduced in 100% of animals compared with that apparent in the vehicle-treated eye. The inhibitory effect was dose-dependent, with a maximal inhibition of 39% (P < 0.01) at a dose of 1 nmole. The immunostaining intensity for NF-KB in the retina was reduced by PDTC injections. No side effects by PDTC in the retina were observed by light and electron microscopy. CONCLUSIONS: NF-kappaB activation appears to be required for retinal angiogenesis, given that the administration of PDTC suppressed retinal neovascularization. PDTC may prove beneficial in the treatment of ischemic neovascular diseases such as diabetic retinopathy and retinal vein occlusion.

Electroporation and bleomycin in glaucoma-filtering surgery.

PURPOSE: To develop a new treatment in glaucoma-filtering surgery using combined treatment with electroporation and the antiproliferative drug bleomycin. METHODS: Pigmented rabbits were treated with both bleomycin (10 micrograms/ml) and localized electric pulses (EP) using a special probe (5 V/cm, 100 msec, 8 pulses) (n = 10; group A). After EP treatment, bleomycin was washed out with 50 ml balanced salt solution, and then a posterior lip sclerectomy was performed on the same area. We also studied rabbits undergoing a posterior lip sclerectomy with bleomycin treatment alone (n = 10; group B), a posterior lip sclerectomy with EP treatment alone (n = 5; group C) and a posterior lip sclerectomy alone using the same operation (n = 5; group D, negative control). The intraocular pressure (IOP) was measured before and 1, 3, 5, 7, 10, 15, and 20 days after surgery. The formation of blebs, the conjunctiva, and the cornea were periodically examined by slit-lamp biomicrography. RESULTS: In every group, the IOP decreased until day 7, and no significant difference was observed among the four groups. In groups B, C, and D (control), the IOP increased gradually from day 10 and thereafter returned to the preoperative level after 15 days. However, in group A, the IOP remained lower than the preoperative level for 20 days; it was also significantly lower than each of the other three groups (P < 0.01). The survival rate of a filtering bleb was significantly higher in group A than in groups B, C, or D, but the survival rate in group B was not higher than groups C or D. No adverse effects were clinically observed in the ocular tissue, such as the cornea and conjunctiva. CONCLUSIONS: The combined treatment with EP and bleomycin was found to decrease IOP more prominently than EP or bleomycin treatment alone in filtering surgery. This treatment thus makes filtering surgery effective by decreasing the dose of the antiproliferative drug and by possibly localizing the drug delivery.

Inhibition of corneal inflammation by the topical use of Ras farnesyltransferase inhibitors: selective inhibition of macrophage localization.

PURPOSE: Ras farnesyltransferase inhibitors are known to block the membrane translocalization of oncogenic Ras protein. They inhibit the cytoplasmic mitogen-activated protein kinase signaling cascade related to Ras protein. Thus far, Ras farnesyltransferase inhibitors have been exclusively regarded with the anticancer drugs. The object of this study was to elucidate the role of Ras farnesyltransferase inhibitors on the corneal opacity induced by an inflammatory stimulus. METHODS: We used a cauterization-induced corneal inflammation model. The central corneas of BALB/c mice were cauterized with silver nitrate (1 mm in diameter). Ras farnesyltransferase inhibitors, either manumycin or gliotoxin eye drops (each drug dissolved in balanced salt solution [BSS] at concentrations of 1 mM), were topically delivered to the cauterized cornea every 8 hours; BSS eye drops were used as a control. Clinical signs such as corneal edema, opacity, and corneal neovascularization, which are major causes of visual disturbance, were then examined 96 hours after the cauterization. The corneal edema and opacity were clinically scored under a stereoscopic microscope. The corneal neovascularization was evaluated by the length of the blood vessels from the limbus and the sum of extension central angle of vascularized limbus. Furthermore, the corneas were examined histologically, and the phenotypes of the cornea-infiltrating cells were analyzed by flow cytometry. RESULTS: The control corneas showed prominent edema, neovascularization, and opacity. Histologic analysis revealed corneal epithelial and endothelial cell loss and a large amount of inflammatory cell infiltration into the corneal stroma. Flow cytometric analysis revealed that most of the infiltrating cells were neutrophils and macrophages. In contrast, the degree of corneal edema, neovascularization, and opacity was significantly less in the manumycin- or gliotoxin-treated corneas than in the control corneas. Histologically, the manumycin- and gliotoxin-treated corneas showed minimum edema and good epithelialization. Flow cytometric analysis showed corneal infiltration of macrophages to be selectively and clearly inhibited. Neither manumycin nor gliotoxin produced any side effects in the noncauterized normal cornea either clinically or histologically. CONCLUSIONS: Ras proteins play an important role in cauterization-induced corneal inflammation and the opacity it induces. Ras farnesyltransferase inhibitors thus have a great potential for improving the treatment of corneal opacity induced by a corneal inflammatory stimulus.

Gene transfer of a soluble receptor of VEGF inhibits the growth of experimental eyelid malignant melanoma.

PURPOSE: To determine the effect of adenovirus-mediated gene transfer of a soluble receptor of vascular endothelial growth factor (VEGF) on the growth of experimental eyelid malignant melanoma. METHODS: An adenovirus vector encoding a soluble VEGF receptor/flt-1 (Adflt-ExR) was constructed. The bovine retinal endothelial cells (ECs) were incubated in a culture medium of 293E1 cells infected by means of an adenovirus vector or uninfected (control), which contained human recombinant VEGF, and the [3H]thymidine uptake was tested. The experimental eyelid malignant melanoma was induced by the injection of B16 melanoma cells (4 x 10(6) cells) into the right upper eyelid of BALB/c nu/nu mice, and the size of the tumor was recorded for 3 weeks after tumor cell injection. The effect of Adflt-ExR was examined in three ways. Model 1: B16 cells were infected by Adflt-ExR beforehand (at a multiplicity of infection [MOI] of 10) and injected into the eyelid. Model 2: Adflt-ExR was injected into pre-established B16 cell-induced eyelid malignant melanoma. Model 3: Adflt-ExR was injected into the femoral muscle of mice before B16 cell injection into the eyelid, and the remote effect was evaluated. An adenovirus vector bearing the LacZ gene (AdLacZ) or phosphate-buffered saline was used as a control. The amount of VEGF and the flt-ExR protein was measured by sandwich enzyme-linked immunosorbent assay (ELISA). Vascularization was evaluated by counting the number and the size of the vessels. RESULTS: The supernatant of Adflt-ExR-transfected cells clearly inhibited VEGF-induced bovine retinal EC proliferation in vitro. In models 1 and 2, the tumor growth in Adflt-ExR-treated mice was significantly lower than that of controls (P < 0.05). In model 3, no significant difference was found (P = 0.14). The molar ratio of VEGF/flt-ExR protein was clearly low in the tumors of Adflt-ExR-treated mice in models 1 and 2 (P < 0.01) but not in model 3 (P > 0.05). In vessel density, the tumors in Adflt-ExR-treated mice had fewer vessels than tumors in control animals in models 1 and 2 (P < 0.05). CONCLUSIONS: Adenovirus-mediated gene transfer of a soluble form of VEGF receptor (flt-1) gene inhibited the growth of the experimental eyelid malignant melanoma. This method may be useful as an antiangiogenic therapy for eyelid malignant melanoma.

Role of cyclic AMP-induced Cl conductance in aqueous humour formation by the dog ciliary epithelium.

1. The effects of isoprenaline, a forskolin derivative NKH-477, and dibutyryl cyclic AMP (db cyclic AMP) on the membrane potential, conductance and cell volume of the dog non-pigmented ciliary epithelium (NPE) were investigated by intracellular potential recording, nystatin-perforated patch clamp technique and videomicroscopic cytometry. 2. The resting membrane potential of NPE was about -70 mV in physiological saline and was depolarized by isoprenaline in a dose-dependent manner with an ED50 of about 3 nM. This depolarization was competitively antagonized by the beta-adrenoceptor antagonist, timolol (pA2 = ca. 9) and almost completely blocked by the Cl transport blocker, DIDS. 3. In single dissociated NPE cells, 10 microM isoprenaline induced an inward current and caused a concomitant decrease in cell volume. The reversal potential measurement indicated that this inward current was carried mainly by Cl ion. DIDS (10 microM) abolished both the current and cell volume decrease. 4. NKH-477 (10 microM) or db cyclic AMP (1 mM) also induced an inward current together with a cell volume decrease, the properties of which were similar to those caused by isoprenaline. 5. These results suggest that beta-adrenoceptor stimulation in NPE leads to an increased rate of aqueous humour production by increasing Cl- efflux via an elevation of cyclic AMP and this effect is efficiently blocked by timolol.

Suppression of infectious virus spread and corneal opacification by the combined use of recombinant interferon beta and interleukin-10 following corneal infection with herpes simplex virus-1 in mice.

The effects of Interleukin-10 (IL-10) and recombinant murine interferon-beta (rMuIFN-beta) on experimental corneal herpes simplex virus 1 (HSV-1) inoculation in BALB/c mice were examined. The mice were inoculated with the HSV-1 strain KOS at their corneas after abrasion. IL-10 was then administered topically once a day for 10 days beginning 2 days post inoculation, while rMuIFN-beta was administered once a day for 10 days beginning 1 day post inoculation. The local viral growth in the inoculated eyes and trigeminal ganglia was reduced in the rMuIFN-beta-treated mice but not in the IL-10-treated mice. In the mice treated with both rMuIFN-beta and IL-10, the degree of both the local viral growth and corneal opacification decreased. The establishment of HSV-1 latency in the trigeminal ganglia was partially prevented by rMuIFN-beta treatment but not by IL-10 treatment. The combined use of the cytokines resulted in both the suppression of viral spread and the prevention of corneal inflammation induced by HSV-1 infection.

Necrotic changes of choroidal melanocytes in sympathetic ophthalmia.

There are various theories as to the origin of epithelioid cells in the choroid with sympathetic ophthalmia. Some investigators propose a transformation of choroidal melanocytes as the origin, and others suggest a histiomonocytic derivation. One reason this controversy exists may be the relative lack of investigations into necrotic changes of choroidal melanocytes. The structural alterations in the choroidal melanocytes of an injured eye with sympathetic ophthalmia were studied, and the sequence of events involved in degeneration and necrotic changes were elucidated. The damaged melanocytes developed vacuolation, and the melanin granules were gathered into autophagosomes or had disappeared. The nuclei of the severely damaged melanocytes became pyknotic. Degenerated cell nuclei were phagocytized by macrophages. It is concluded that choroidal melanocytes may not transform into epithelioid cells and that they disappear from the choroid following degeneration.

Class II major histocompatibility complex on melanocytes of Vogt-Koyanagi-Harada disease.

The eyes obtained from two Japanese autopsy cases of patients with Vogt-Koyanagi-Harada disease were immunohistochemically examined. Both patients, a 63-year-old woman and a 68-year-old man, were clinically and histologically diagnosed as having Vogt-Koyanagi-Harada disease. Immunohistochemically, the choroidal infiltrate was composed predominantly of T lymphocytes with a larger proportion of helper/inducer T cells than suppressor/cytotoxic T cells and it also included activated lymphocytes expressing CD26 and CD25 antigens. Class II major histocompatibility complex was expressed in the choroidal melanocytes as well as in the endothelium of the choriocapillaris. Depositions of complement, however, were focally noticed in the choroid. Our results indicate that the cell-mediated immune process plays an important role in the development and progression of Vogt-Koyanagi-Harada disease, while choroidal melanocytes appear to play a pathogenic role in this disease.

Epiretinal membrane formation. Light and electron microscopic study in an experimental rabbit model.

OBJECTIVE: To clarify the role of retinal glial cells in epiretinal membrane formation. METHODS: We injected autologous whole blood into the vitreous cavity of albino rabbits and studied the events in the vitreoretinal interface at intervals during the course of 1 year by light and electron microscopy. RESULTS: Epiretinal membranes were first found 2 weeks after the treatment. At this stage, epiretinal membranes were composed of both glial cells and macrophages. Mitotic figures of glial cells were found in the retina. The nuclei of glial cells migrated, passing through the inner limiting membrane and onto the retinal surface. At 6 months, macrophages and red blood cells disappeared from the epiretinal membranes. The epiretinal membranes became thicker with time. Finally, these epiretinal membranes were composed solely of glial cells. CONCLUSIONS: At the early stage, macrophages participate with glial cells in epiretinal membrane formation; however, glial cells are the main constituent of epiretinal membranes during the late stage.