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Biomedical subjects

K Akeo

Publications and source records attributed to K Akeo.

At least 37 records · Page 2Linked to original sources
PubMed1995

[Electron microscopic studies of retinal pigment epithelial cells incubated with L-dopa in 10% and 20% oxygen].

In order to make clear how the ultrastructure of the bovine second-passage retinal pigment epithelium (RPE) was affected by exposure to L-dopa and oxygen, we compared the toxic effects of melanotic and amelanotic RPE. The oxygen levels in the incubators and the L-dopa concentrations in the media were adjusted to 10% or 20% oxygen by adding nitrogen gas and 100 microM or 250 microM L-dopa, respectively. After 24 hours, melanotic and amelanotic RPE were fixed and embedded in epon. Ultrathin sections were examined by transmission electron microscopy. Rough endoplasmic reticula were found to be dilated and ribosomes were extinguished in both melanotic and amelanotic RPE. The damage by L-dopa in melanotic RPE was less than that in amelanotic RPE and less in 10% oxygen than in 20% oxygen.

Animals↗
PubMed1994

A comparison between melanotic and amelanotic retinal pigment epithelial cells in vitro concerning the effects of L-dopa and oxygen on cell cycle.

Melanin precursors and free radicals, cytotoxic substances, are produced during melanin synthesis by tyrosinase. We compared these cytotoxic effects of L-dopa and oxygen on the cell cycle of melanotic retinal pigment epithelial (RPE) cells with amelanotic RPE cells because of the differences of tyrosinase activities between melanotic and amelanotic RPE cells. Flow cytometric DNA analysis of RPE cells exposed to L-dopa (100 microM and 250 microM) were conducted at several oxygen concentrations (20%, 10%, and 5%). The dose-dependent effect of L-dopa to arrest the cell cycle (the S phase) was more pronounced in melanotic than in amelanotic RPE cells, and oxygen caused arrest in the G1 phase.

Animals↗
PubMed1994

Autosomal dominant retinitis pigmentosa. A mutation in codon 181 (Glu-->Lys) of the rhodopsin gene in a Japanese family.

The PCR/restriction endonuclease digestion (RE) assay and PCR/SSCP analysis of the rhodopsin gene in 13 Japanese families with autosomal dominant retinitis pigmentosa (ad RP) revealed a G-A substitution of the first nucleotide of codon 181, replacing Glu (GAG) with Lys (AAG), in one family. The proband showed an early onset of symptoms in childhood with a diffuse loss of rod and cone function and a relatively good preservation of cone function, corresponding to the type with relatively rapid progression to blindness (type I category of ad RP).

Adult↗
PubMed1993

A novel Cys-214-Ser mutation in the peripherin/RDS gene in a Japanese family with autosomal dominant retinitis pigmentosa.

We have screened for possible disease-causing mutations in the peripherin/retinal degeneration slow (RDS) gene in 13 Japanese families with autosomal dominant retinitis pigmentosa (ADRP). Using polymerase chain reaction-single strand conformation polymorphism analysis, a novel mutation at codon 214 was found in which the highly conserved cysteine was replaced with a serine in one family. The mutation at codon 214 was found in all three affected siblings of this family, but none of the 40 normal control individuals had this mutation. These results strongly suggest that the mutation is pathogenic for RP in this family. The clinical phenotype for this family is a late-onset form of ADRP.

Age of Onset↗
PubMed1992

The effect of oxygen on melanin precursors released from retinal pigment epithelial cells in vitro.

The autoxidation of dopa to melanin in culture media causes toxicity to retinal pigment epithelial (RPE) cells and endothelial cells. The damage is specific to cell type and to the ambient oxygen concentration. To determine whether RPE cells influence the oxidation of dopa to media, we compared light absorbing dopa derivatives in the media exposed to cells with those found in the media incubated without cells. Dopa was extensively oxidized in the presence of RPE cells, and more light absorbing substances were generated with higher dopa and oxygen concentrations. However, an increase in ambient oxygen concentration decreased the quantity of several dopa derivatives which had been formed. The data provided evidence that RPE modulated dopa metabolism. Quinolic derivatives produced from a tyrosinase reaction and dopa-melanin formation moved the peak absorbance wavelength of dopa into the visible range. The spectrum between the dopa-derived compounds in the media has an absorbance at 240-275 nm and a maximum around 300 nm with a shoulder near 375 nm. Gaussian analysis (peak separation) resolved these spectra into five components: a sharp band at 248 nm, a band at 295 nm, a large band at 359 nm, and two broad bands at 459 and 585 nm.

Animals↗
PubMed1992

Comparison of effects of oxygen and antioxidative enzymes on cell growth between retinal pigment epithelial cells and vascular endothelial cells in vitro.

We assayed the proliferation of porcine retinal pigment epithelial (RPE) cells, bovine melanotic and amelanotic RPE cells, and bovine aortic endothelial cells exposed to 20, 10 and 5% oxygen and compared their responses to oxygen and antioxidative enzymes (superoxide dismutase and catalase). Irrespective of the cell type, the cell growth was optimal in 10% oxygen that is most closely approximating to the oxygen concentration prevailing in the cellular environment of the choroid and the retina in vivo. However, the effects of oxygen concentrations were cell specific because bovine endothelial cells were influenced by lowering of oxygen concentrations more significantly than bovine and porcine RPE cells. Moreover, addition of antioxidative enzymes caused significant improvement in growth of porcine RPE cells, but had no significant effects on bovine RPE cells. On the contrary, the bovine vascular endothelial cells represented the only one cell type significantly inhibited by antioxidative enzymes, i.e., a decrease in reactive intermediates of oxygen was seen in the media. Our results show that responses of vascular endothelial cells to reactive species of oxygen were distinctly different from those of RPE cells and more easily influenced by the environment related to hypoxia than RPE cells.

Animals↗
PubMed1990

Growth of cultured RPE and endothelial cells is inhibited by blue light but not green or red light.

Blue light, but not green or red light, inhibited growth of retinal pigment epithelial (RPE) cells, aortic endothelial cells, and fibroblasts in vitro. Significant inhibition was observed in all 3 cell types exposed for 18 hr to blue light (425-500 nm) at 42 J/cm2. Damage was prevented by inclusion of superoxide dismutase (SOD) and catalase, providing evidence for a photooxidative mechanism. Dopa (100 microM) also caused oxidative damage that suppressed growth of all 3 cell types. A synergism of dopa and light effects was observed in endothelial cells and fibroblasts, but the agents caused additive effects on RPE cells. Endothelial cells were the most sensitive to dopa, light, and the two combined. Fibroblasts were the only cell type that exhibited greater sensitivity to light than to dopa. These data suggest that oxygen-mediated damage to the growing blood vessels in the retina of a premature infant may be exacerbated by exposure to blue light. A further implication is that restriction of RPE melanogenesis to the prenatal period of darkness and lower oxygen protects the retina from simultaneous oxidative challenge by light and by reactive species generated during oxidation of dopa released to the extracellular environment.

Analysis of Variance↗
PubMed1989

Dopa and oxygen inhibit proliferation of retinal pigment epithelial cells, fibroblasts and endothelial cells in vitro.

Some quinones catalyze superoxide formation in a futile cycle involving electron transfer to molecular oxygen. If the quinolic precursors of melanin participated in the futile cycle, the high ambient oxygen surrounding postnatal RPE would make continued melanogenesis risk for the retina. To probe the possibility that arrest of melanogenesis in postnatal RPE is a protective mechanism, we assayed the growth rates of RPE cells, aortic endothelial cells and skin fibroblasts exposed to 0, 10, 50, 100 and 250 microM dopa. To assess the contribution of the futile cycle, we studied the effects of oxygen concentration and the antioxidants, superoxide dismutase and catalase. We found that all three cell types were significantly and dose-dependently inhibited by dopa and that the effects of dopa were oxygen dependent. The powerful inhibition of RPE cells by dopa was counteracted by inclusion of superoxide dismutase and catalase, but not by an inhibitor of dopa oxidase (phenylthiourea), indicating that the mechanism of growth suppression did not involve melanogenesis but, rather, dopa-dependent formation of superoxide in the media. Endothelial cells were more sensitive to the dopa-mediated oxidative damage than were RPE cells or fibroblasts. Fibroblasts were most affected by oxygen alone, and least affected by dopa. These data suggest that suppression of melanogenesis in the postnatal RPE may be an important mechanism for preventing oxidative damage to the retina and the choriocapillaris We propose that the generation of oxygen radicals by the quinone futile cycle is a viable model of the damage of cells in culture by dopa.

Animals↗
PubMed1988

Electron microscopic studies of the endocytotic process of cationized ferritin in cultured human retinal pigment epithelial cells.

The endocytotic process in cultured human RPE cells was observed after 1 min, 20 min, and 2 h incubation with cationized ferritin. Within 1 min the ferritin particles were seen to attach to the cell membrane, especially between microvilli. Uncoated and coated pits could be recognized on the cell membranes, and uncoated and coated endocytotic vesicles were found in the cytoplasm after 20 min of incubation. These vesicles were surrounded by abundant microfilaments and had no visible membranes. Loss of membrane may be an initial step in the process of developing into the irregular clumps of ferritin particles found inside the plasma membrane. With time, more irregular clumps of ferritin, smaller than the particles introduced during incubation, appeared just beneath the cell membrane. Lysosomes were adjacent to the clumps of ferritin particles associated with microtubules and finally degraded these particles. The phagolysosomes containing many particles were surrounded by many microtubules. Small ferritin particles surrounded but had not entered the rough endoplasmic reticulums, and no particles were seen either around or in the Golgi apparatus.

Cations↗
PubMed1988

Electron microscopic comparative studies of phagocytic processes between outer segments and latex microspheres in cultured human retinal pigment epithelial cells.

The process of phagocytosis in cultured human retinal pigment epithelium (RPE) cells was observed after more than 2 h of incubation with human outer segments and latex microspheres. Fingerlike microvilli were attached to outer segments and entwined around both its ends. The microvilli enveloped the outer segment and cut into the membranous structure, and the same process as that seen in in vivo shedding was observed. The looplike disk membranes and the whole of the outer segment were ingested into the cytoplasm and degraded by the lysosome. Latex microspheres were also ingested into the cytoplasm so as to be enveloped in many fingerlike microvilli. Microfilaments were concentrated in the vicinity of latex microspheres and outer segments, and latex microspheres were placed between two microtubules. Furthermore, when latex microspheres and outer segments were ingested into the cytoplasm, the dilated rough endoplasmic reticulum (rER) contained materials of high electron density and attached ribosomes increased. The rER of the cultured human RPE cells seemed to show a high level of protein synthesis during the phagocytic process. It was observed that cytoskeletons, such as microfilaments and microtubules, and lysosomes had important functions in the phagocytic process, and that there were basically no differences among the objects phagocytized by the cultured RPE cells.

Cells, Cultured↗
PubMed1988

Superoxide dismutase activity and growth of retinal pigment epithelial cells are suppressed by 20% oxygen in vitro.

Despite knowledge of the toxicity of oxygen to the retina, its effects on the retinal pigment epithelium have not been considered. We examined the effect of 20%, 10% and 5% oxygen on growth and superoxide dismutase (SOD) activity of porcine retinal pigment epithelial cells (RPE). Growth of RPE cells was very significantly lower in 20% oxygen than in either 10% or 5%; optimal growth occurred at 10% oxygen, the concentration most like their environment in vivo. Inclusion of SOD and catalase in the media very significantly stimulated growth in 20% oxygen. The SOD activity of RPE cells was significantly related to ambient oxygen. In first passage (P1) cells, SOD activity was 44% lower on day 7 than on day 1 of culture in 20% oxygen (p less than or equal to 0.05). Transfer of cells growing in 20% oxygen to 5% oxygen arrested the decrease in SOD and resulted in significantly higher SOD levels. In fourth passage (P4) cells grown in 20% oxygen, SOD was 25% and 44% lower than cells in 10% and 5% oxygen, respectively. After one week, SOD levels in the P4 cells were significantly higher than in P1. A statistical model of SOD activity in RPE cells indicated significant negative correlations with both oxygen concentration and the cell number. Growth of RPE cells was significantly influenced by oxygen level, days of culture and passage number, but not SOD activity. We conclude that traditional culture conditions support generation of free radicals in tissue culture media that suppress both growth and superoxide dismutase activity.

Catalase↗
PubMed1983

Acute retinal necrosis (Kirisawa's uveitis).

Four cases of acute retinal necrosis accompanied by acute uveitis, retinal obliterative vasculitis and confluent yellow-white lesions in the peripheral fundus followed by retinal detachment are reported. The clinical course was rapidly progressive and the prognosis was poor despite various treatments. No systemic abnormality was found and etiology could not be identified. In reviewing the 40 cases so far reported, including our 4 cases, we came to the conclusion that acute retinal necrosis and Kirisawa's uveitis fall into the same clinical study.

Adolescent↗
PubMedDate not supplied

Multicenter genetic study of retinitis pigmentosa in Japan: I. Genetic heterogeneity in typical retinitis pigmentosa.

A nationwide, multicenter study of typical retinitis pigmentosa (RP) was carried out in collaboration with 18 hospitals throughout Japan to obtain current information for genetic counseling. We analyzed the genetic heterogeneity of RP based on the parental consanguinity of 434 probands registered during a 6-month period in 1990. A gradual decline in the frequency of consanguineous marriage was recognized among the normal parents of RP patients. The relative frequencies of inheritance patterns were estimated as: autosomal recessive, 25.2%; autosomal dominant, 16.9%; X-linked, 1.6%; and simplex, 56.3%. A comparison of these results with previous reports in Japan revealed a decline in the relative frequency of autosomal recessive cases and an increase in simplex cases. This suggests a decrease in the incidence of autosomal recessive retinitis pigmentosa in Japan, as well as the necessity for exhaustive investigations aimed at identifying inheritance patterns for RP patients seeking genetic counseling.

Adolescent↗