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Biomedical subjects

S Okisaka

Publications and source records attributed to S Okisaka.

At least 37 records · Page 2Linked to original sources

X-ray microanalysis and phagocytotic activity of cultured retinal pigment epithelial cells in hypoxia.

PURPOSE: To investigate the influence of the functional and morphological changes induced in retinal pigment epithelial (RPE) cells by retinal ischemia, we evaluated the phagocytotic activity, the concentration of various elements, and ultrastructure in cultured RPE cells in hypoxia. METHODS: The concentrations of oxygen in incubators were adjusted to 20, 10, and 1% by the addition of nitrogen for 72 hr. To observe phagocytotic activity and its relationship to actin filaments, the filaments of RPE cells incubated with fluoresbrite carboxylate YG microspheres were stained with rhodamine phalloidin. Some of the specimens were subjected to X-ray microanalysis by scanning electron microscope after being fixed, freeze-dried, and coated with carbon to investigate the cytoplasmic concentration of elements. A part of the latter specimens was also observed by transmission electron microscope after being embedded in epon and cut into ultrathin sections to see the ultrastructural changes inside cell. RESULTS: Lowering oxygen concentrations from 20% to 1% swelled RPE cells and decreased the number of fluoresbrite carboxylate YG microspheres phagocytized by RPE cells. Phagocytosis of a large amount of latex beads (30 microl) for 24 hr in 1% oxygen caused a disruption of RPE cells. Na, S, and P were detected in RPE cells cultured in 20% oxygen. Reducing the oxygen concentration from 20 to 10 or 1% significantly decreased Na and increased S. Mitochondria were observed in RPE cells in 20 and 10% oxygen, but many vacuoles were observed in the cytoplasm in 1% oxygen. CONCLUSION: Hypoxia as low as 1% oxygen induced malfunction of phagocytosis and the fragility of RPE cells. We could speculate the imbalance of the electrolytes such as Na or a decrease of antioxidants such as glutathione containing S as a reason of disturbance of cell viability.

Actins↗

[A case of primary orbital chondrosarcoma].

The case of a twenty-year-old male with orbital chondrosarcoma is reported. He visited National Defense Medical College Hospital because of reduced vision in the right eye since two months previously. His corrected visual acuity was 8/20 in the right eye and 20/20 in the left eye. Fifteen degrees lateral displacement of the right globe and limitation of right ocular movement were recognized. Right fundus examination revealed optic disc edema and protuberant nasal fundus. Orbital computed tomography (CT) demonstrated a high density area between the inner part of the right orbit and the ethmoid sinus. Magnetic resonance imaging (MRI) showed a smoothly outlined and low intensity (T1) space occupying lesion. This lesion was irregularly enhanced by gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA). This orbital tumor was removed by an anterior approach. Histopathological examination revealed well-differentiated chondrosarcoma (grade 1) as determined by small prominent chondromatous cell projection into the collagen fibrous stroma, and existence of binucleate cells in the hypercellular region. After the operation the disc edema disappeared and his corrected right visual acuity improved to 20/20.

Adult↗

[A case of dermatomyositis with severe retinopathy in a patient who died of acute interstitial pneumonia].

Sight threatening ocular complications are rare in adult patients with dermatomyositis. We encountered a 52-year-old female with dermatomyositis who had severe visual disturbance and rapidly progressive intersitial pneumonia. She was admitted to our hospital because of skin erythema, general fatigue, mild fever, and severe bilateral visual disturbance. Rentinal hemorrhages, cotton wool spots, and macular edema were observed in her fundus at the first ophthalmic examination. A diagnosis of dermatomyositis was made because of the myogenic pattern of her electromyogram, elevation of serum creatine kinase, and skin lesions. Oral prednisolone treatment was started and the retinopathy was improved, but was complicated by acute interstitial pneumonia. The interstitial pneumonia was not respond to steroid pulse therapy with methylprednisolone, and the patient died of respiratory failure on the 47th day after the onset of visual symptoms. In adult dermatomyositis patients, the complication of severe retinopathy should be considered as a risk factor for rapid progress of interstitial pneumonia.

Acute Disease↗

[Pterygium and mast cells--mast cell number, phenotype, and localization of stem cell factor].

We examined the number and phenotype of mast cells, and the localization of stem cell factor (SCF) as a growth factor of mast cells in the excised tissue of 38 cases of pterygium. In histopathology with toluidine blue stain and immunohistochemistry with a monoclonal antibody to tryptase, the mean mast cell count in pterygium specimens was twice as high as in normal conjunctiva. In pterygium specimens more than 94% of tryptase-positive mast cells were found to express chymase and c-kit. There was no phenotypic difference between mast cells in pterygium and normal conjunctiva. In all immunohistochemical specimens in which we could examine the head of the pterygium, SCF was expressed in subepithelial fibroblasts at the central edge of pterygium. The results suggest that overexpression of SCF was accompanied with the augmentation of mast cells in the pterygium.

Cell Count↗

Histology of fetal eyes with oculocutaneous albinism.

The diagnosis of tyrosinase-negative oculocutaneous albinism (OCA) was made in a 19-week-old fetus by skin biopsy. Because the parents had an 11-year-old son with tyrosinase-negative OCA, they requested that the fetus be aborted at the 20th week of gestation. A histological analysis of the eyes was performed. Throughout the retina, the ganglion cell layer was separated from the inner neuroblastic layer by the inner plexiform layer. However, the number of ganglion cells was decreased and the nerve fiber layer was immature. Bipolar and horizontal cells had begun to segregate into the inner nuclear layer. Rods and cones were identifiable in the posterior, but not peripheral, retina. Cones were more numerous in the center of the retina, and no rod-free area was identifiable. In addition, the ciliary body (epithelial folds, blood vessels in the mesodermal connective tissue core, and ciliary muscle) was less developed than in a normal fetus. Melanosomes in the retinal pigment epithelium only contained filaments without melanization and were therefore classified as stage I or II melanosomes. However, the ciliary epithelium also contained some stage III melanosomes with melanin adherent to the filaments.

Abortion, Therapeutic↗

Effects of superoxide dismutase and catalase on growth of retinal pigment epithelial cells in vitro following addition of linoleic acid or linoleic acid hydroperoxide.

The rod outer segments of the retina that are phagocytized by retinal pigment epithelial (RPE) cells are susceptible to lipid peroxidation because of their high content of polyunsaturated fatty acids. Linoleic hydroperoxides (LHP), synthesized by peroxidation of linoleic acids (LA), produce greater damage to retinal function than does LA. We compared the effects of LHP and LA on the growth of cultured chick embryonic RPE cells and analyzed a model of data sets using multiple linear regression for the number of cells as a function of time. The spectrum of LA had a sharp peak at 205 nm and a broad spectrum at 235 nm, while LHP had only a broad spectrum at 235 nm. Exposure to LA and LHP caused dose-dependent damage of chick embryonic RPE cells: they were significantly more affected by the addition of LHP than LA. The antioxidative enzymes catalase and superoxide dismutase minimized damage to the RPE cells caused by LHP in proportion to the enzyme concentration. However, RPE cells incubated with LA were more affected by the enzymes than when no enzymes were added.

Animals↗

Comparative effects of linoleic acid and linoleic acid hydroperoxide on growth and morphology of bovine retinal pigment epithelial cells in vitro.

PURPOSE: Outer segments of the photoreceptor rods that are phagocytized by the retinal pigment epithelial (RPE) cells contain a high proportion of polyunsaturated fatty acids (PUFA). PUFA are susceptible to lipid peroxidation. We hypothesized that the resulting peroxides could injure RPE cells leading to retinal degeneration. Accordingly, we compared the effects of linoleic acid (LA) and its hydroperoxide (LHP) on the growth and morphology of RPE cells using laser scanning microscopy and transmission microscopy. METHODS: We counted the number of RPE cells after incubation for 24 and 48 hrs with concentrations of LA or LHP of 0.035, 0.175, and 0.35 mM. To observe the actin filaments, cultured RPE cells were stained with rhodamine phalloidin. The cells were prefixed with 2% glutaraldehyde and postfixed in 1% osmium tetroxide. Specimens were embedded in Epon 812 after dehydration, and the ultrathin sections were doubly stained with 2% uranyl acetate and 2% lead acetate for examination by transmission electron microscopy. RESULTS: Exposure to LA or LHP produced dose-dependent damage to RPE cells with a significantly greater effects of LHP than LA. After incubation for 24 hrs with 0.35 mM LA, the number of vacuoles in RPE cells exceeded that observed in control RPE cells by 365 nm laser microscopy. Exposure to 0.35 mM LHP for 24 hrs produced a pycnotic nucleus, with diffuse and granular autofluorescences observed in and around it. Exposure of RPE cells to 0.35 mM LA for 24 hrs showed that the LA incorporated into the lysosomes was digested and released extracellularly from lysosomes via exocytotic vesicles. However, such exposure to LHP damaged the RPE cells, including the membranes in the pinocytotic vesicles. The packed membranes resembled myelin. CONCLUSIONS: While the LA incorporated into the lysosomes was released extracellularly, LHP persisted in the RPE cells, being observed as autofluorescent lipofuscin-like materials. LHP was cytotoxic, and caused damage to the membranes of pinocytotic vesicles and lysosomes.

Animals↗

[Aging changes in conjunctiva visualized by fluorescein angiography and histopathology].

In order to investigate the aging changes of the conjunctiva, we examined fluorescein-positive areas by fluorescein angiography. The deposition of melanin in basal cells, the degranulation of mast cells, and the lumens of capillaries were examined by light microscopy in 35 cataractous eyes without diabetes or hypertension. The fluorescein leakage increased with aging. The number of basal cells with deposition of melanin granules and mast cells with degranulation correlated significantly with aging. On the other hand, the narrowing of capillary lumens in conjunctival stroma was not observed with aging. Based on the above results, we conclude that the weakness of the conjunctival vessels without the narrowing of capillary lumens might be caused by functional disorder of the endothelium and pericytes. The increase of deposition of melanin granules in basal cells may be caused by the breakdown of the metabolic pathway of melanin. Long-time mechanical stimuli and metabolic disorders of active substances may increase the degranulation of mast cells.

Adult↗

[Immunohistochemical localization of acidic and basic fibroblast growth factor through corneal neovascularization in vivo and in vitro].

Neovascularization on the center of rabbit cornea was induced by 5N.NaOH alkali burns. We studied the change in localization of acidic fibroblast growth factor (a-FGF) and basic fibroblast growth factor (b-FGF) through corneal neovascularization with immunohistochemistry, using eyes which we enucleated on the 3rd, 7th, and 14th day. Moreover, by co-cultivation of rabbit corneal stromal cells and adrenal cortical vascular endothelium of bovine, a capillary-like code was induced, in which we also studied the localization of a-FGF and b-FGF on the 3rd, 7th, and 14th day. On the 14th day after the alkali burn we recognized intrastromal neovascularization and positive staining of a-FGF and b-FGF around it. Strong staining of a-FGF was observed in goblet cells through the experimental period. In control eyes we recognized positive reaction of a-FGF in corneal and conjunctival epithelium. In co-cultured cells, we recognized positive staining of both a-FGF and b-FGF around the capillary-like code which was induced among the corneal stromal cells.

Animals↗

[Natural process of wound healing of photocoagulated retinal pigment epithelium in culture--observation of DNA synthesis by BrDu incorporation].

We examined the proliferation of retinal pigment epithelial (RPE) cells after krypton laser photocoagulation in culture. A pigmented monolayer of chick embryonic RPE cells was cultured on a collagen membrane placed on collagen gel. RPE cells were labeled with bromodeoxyuridine (BrDu) every 12 hours until 7 1/2 days after the photocoagulation and stained immunocytochemically with anti BrDu antibody. Immediately after the photocoagulation, RPE cells became detached at the burned lesion and the collagen membrane beneath the RPE layer was exposed. Some cells adjacent to the burned lesion showed DNA synthesis and subsequent mitosis between 12 to 24 hours after the photocoagulation. Cells with labeled nuclei migrated into the denuded burned area after 24 hours and covered the whole burned area within three days after the photocoagulation. DNA synthesis continued in these on the burned lesion after complete coverage of the lesion but stopped temporarily 3 1/2 to 4 1/2 days after the photocoagulation. Thereafter DNA synthesis increased again and continued until the end of the experiment. Such use of the cultured RPE cells might be useful in studying cellular reaction after photocoagulation.

Animals↗

[Comparative study on diode laser (680 nm) and krypton laser mild photocoagulation--histological findings and therapeutic outcome for diabetic maculopathy].

A histopathological study of macaca monkey retina-choroid was carried out for one month after mild photocoagulation with a diode laser (680 nm) and a krypton laser (647 nm). Both lasers produced the same damage to the outer layer of retina and to the melanocytes of 1/3 of the inner choroid. Mild focal photocoagulation was done for 6 patients (12 eyes) with diabetic maculopathy. The patients were 41 to 67 years old, suffering from diabetes mellitus for 5 to 25 years. The grade of their retinopathy was Fukuda's classification AI, AII, and BI. AII eyes were tested before and 3-6 months after photocoagulation for corrected visual acuity, fluorescein angiography, and visual field sensitivity by Octopus automated static perimetry. Visual acuity was stable in all cases. Central sensitivity was better in 2 eyes and unchanged in 4 eyes in both types of laser photocoagulation. In cases of total loss, one eye was improved, 4 eyes were unchanged and one eye was worsened by diode laser photocoagulation and one eye was improved, 2 eyes were unchanged and 3 eyes were worsened by krypton laser photocoagulation. The diode laser (680 nm) might be as useful for ophthalmic laser treatment as the krypton laser.

Adult↗

Missense mutation of rhodopsin gene codon 15 found in Japanese autosomal dominant retinitis pigmentosa.

Heterozygous missense mutation in codon 15 of the rhodopsin gene was detected in a patient with autosomal dominant retinitis pigmentosa (ADRP), where a transition of adenine to guanine at the second nucleotide in codon 15 (AAT-->AGT), corresponding to a substitution of serine residue for asparagine residue (Asn-15-Ser) was detected. None of the remaining unrelated 42 ADRP, 24 autosomal recessive RP (ARRP) and 34 normal individuals had this alteration. Her funduscopic findings were sectorial in type similar to that of the patients with the same mutation found in an Australian pedigree (Sullivan et al., 1993). This study shows phenotypic similarities in patients with the same mutation of a different ancestry.

Amino Acid Sequence↗

Clinical and histopathologic features of corneal dystrophies in Japan.

PURPOSE: To examine retrospectively the frequency of various corneal dystrophies among Japanese patients who underwent keratoplasty or keratectomy at the authors' institution over a 34-year period, and to compare the histopathologic features of these disorders in the Japanese population with those reported in the Western literature. METHODS: Corneal specimens obtained during keratectomy or keratoplasty (lamellar and penetrating) performed at the authors' institution from 1959 through 1992 were reviewed. Immunohistochemical studies were performed using monoclonal antibodies to keratan sulfate and gelsolin, as well as two lectins (concanavalin A and wheat germ agglutinin). RESULTS: Of 1259 corneal specimens, 159 (12.6%) specimens from a total of 80 patients showed corneal dystrophy. Virtually all were non-Fuchs dystrophies; only one case of primary Fuchs dystrophy was identified histologically. Granular dystrophy and gelatinous drop-like dystrophy were the most common dystrophies identified in the specimens, largely because of multiple specimens from individual patients with recurrent disease. These two disorders accounted for 86 of the 159 specimens. In terms of numbers of patients, lattice dystrophy was the most common (26 patients, 32.5%), followed by macular dystrophy (16 patients, 20%), gelatinous drop-like dystrophy (15 patients, 18.8%), granular dystrophy (14 patients, 17.5%), and Avellino dystrophy (3 patients, 3.75%). Dystrophies represented by only one or two patients included congenital hereditary endothelial dystrophy, primary spheroidal keratopathy, posterior polymorphous dystrophy, Schnyder crystalline dystrophy, and Fuchs dystrophy. CONCLUSIONS: This histopathologic study showed a very low incidence of Fuchs dystrophy in the authors' Japanese patient population, compared with the incidences seen in studies of populations in Western countries. Of the non-Fuchs dystrophies, lattice dystrophy was the most common among the patients, although there were large numbers of specimens with granular dystrophy and gelatinous drop-like dystrophy due to their recurrent character. The causes of clinical and histopathologic differences and similarities among the Japanese patients and the patients described in the Western literature are likely related to genetic factors, but a complete understanding of their specific mechanisms awaits future molecular biologic and genetic elucidation.

Antibodies, Monoclonal↗

[A case of radiation optic neuropathy after resection of a pituitary adenoma].

A case of optic neuropathy after postoperative radiation therapy is reported. A 69-year-old woman had a partial resection of a pituitary adenoma in 1990 and was treated with 45Gy of irradiation to the postoperative pituitary lesion for one month. Seven months later she had sudden right visual field loss. Goldmann perimetry examination revealed remarked visual field defect in her right eye with visual acuity of 1.0. The right relative afferent pupillary defect was positive. The value of critical flicker fusion for her right eye was reduced and the amplitude of steady-state pattern-reversal visually evoked cortical potential was significantly less for the right monocular stimulation than that for the fellow eye stimulation, but Ganzfeld electroretinograms were normal for both eyes. Magnetic resonance imaging using Gadolinium-diethylene triaminepenta-acetic acid revealed enhancement on the right optic nerve, which had not been recognized immediately after the radiation therapy, without any suggestion of right optic nerve compression by the residual pituitary adenoma.

Adenoma↗

[Effects of anticoagulant therapy on retinal vein occlusion].

The combined effect of prostaglandin E1, stellate ganglion block, warfarin potassium and photocoagulation on 66 cases (eyes) of retinal vein occlusion was investigated immediately after, and 3 months and 6 months after final treatment. The time between onset and treatment was 2 months or less. Macular hemorrhage and edema were observed in all cases. The effectiveness rate 6 months after treatment was 35% in hemorrhagic central retinal vein occlusion and 63% in branch retinal vein occlusion. Visual acuity increased gradually with time from immediately after to 6 months after treatment. The effectiveness of treatment including warfarin potassium was lower than that of treatment without warfarin potassium in branch retinal vein occlusion, and many cases showed a marked decrease of visual acuity which might be related to the prolonged hemorrhage. These results suggest that combining prostaglandin E1, stellate ganglion block and photocoagulation in retinal vein occlusion produces better effects on visual acuity than ordinary treatment or spontaneous regression.

Aged↗

[Incidence of tumors and tumor-like lesions in the conjunctiva and the cornea].

Incidence of tumors and tumor-like lesions in conjunctiva and cornea seen at Juntendo University during the 13-year period from 1980 to 1992 was analysed histopathologically. There were 126 cases including 116 benign (92.1%) and 10 malignant lesions (7.9%). Pigmented nevi (32 cases) were most frequently found, accounting for 27.6% of the benign tumors and tumor-like lesions, half of which were compound nevi (16 cases). Nevi were closely followed by cysts (24 cases, 20.7%) and dermoids (15 cases, 12.9%). Carcinoma in situ (3 cases) was most frequently found, accounting for 30% of the malignant tumors (10 cases), closely followed by malignant lymphoma (2 cases), squamous cell carcinoma (2 cases), mucinous carcinoma (1 case), sebaceous gland carcinoma (1 case) and metastatic tumor (1 case). The ratio of males to females was 4:6. Age distribution of the patients was 47-92 years.

Adolescent↗