Biomedical subjects

M Tamai

Publications and source records attributed to M Tamai.

At least 55 records · Page 3Linked to original sources

PubMed2000

Variable R1 region in varicella zoster virus in fulminant type of acute retinal necrosis syndrome.

BACKGROUND/AIMS: Varicella zoster virus (VZV) is a causative agent in acute retinal necrosis (ARN) syndrome. However, in spite of aggressive antiviral therapy, clinical characteristics among patients have varied. Different viral strains were examined to determine their respective role in producing clinical characteristics. The viral strains were also compared with those of previously reported ones. METHODS: To differentiate VZV strains R1 and R5, variable regions of VZV were amplified by nested polymerase chain reaction (PCR) in 11 eyes of 10 patients. Sequence analysis was also performed. RESULTS: Four cases had strains diverted only at the tip of the 3' end of the R1 variable region, similar to that of the H-N3 strain, which was previously reported. Conversely, other cases were diverted to other regions. Interestingly, some of the latter cases showed multiple PCR products in the R1 region that were generated by the truncation of either the 5' or 3' R1 region. Final visual acuities of these patients were less than 0.2. The former cases showed final visual acuities more than 0.4. Only two variants were from the R5 region. No patient had the same viral strain as the European Dumas type. CONCLUSION: These results showed that variable VZV strains participated in ARN. Using PCR of the R1 variable region, it was estimated that patients with a more fulminant type of ARN may have diverse viruses with extensive replication in the affected eyes.

Adolescent↗

PubMed2000

Effect of epidermal growth factor on sodium-dependent L-alanine transport in LLC-PK1 cells.

We evaluated the role of epidermal growth factor (EGF) in the regulation of L-alanine transport in LLC-PK1 renal epithelia. After 2 h of incubation, EGF had no significant effect on L-alanine uptake by LLC-PK1 cells. However, prolonged (16 h) incubation with 2 and 20 ng/ml of EGF resulted in significant increases in sodium-dependent L-alanine uptake as compared with controls. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA; 20 ng/ ml) caused a marked increase in sodium-dependent L-alanine uptake after both 2 and 16 h of incubation, and the treatment with TPA (20 ng/ml) EGF (20 ng/ml) for 16 h resulted in significant acceleration of the TPA-stimulated increase in L-alanine uptake by LLC-PK1 cells. Coincubation with H-7 (20 microM) inhibited both EGF- and TPA-stimulated increases in L-alanine uptake, and genistein (20 microg/ml) blocked the stimulatory effect of EGF in L-alanine transport to the control level. Furthermore, coincubation with cycloheximide (20 microg/ml) for 16 h inhibited both EGF- and TPA-stimulated increases in L-alanine transport to a great extent. The sodium- independent L-alanine uptake was not affected by treatment with either EGF or TPA. These results suggest that the activation of protein kinase C through tyrosine kinase activation plays a role in the EGF effect of stimulating L-alanine transport in LLC-PK1 cells and that the effect is mainly due to increased protein de novo synthesis which occurs after protein kinase C activation.

Alanine↗

PubMed2000

Ultrasound biomicroscopic analysis of the human ciliary body after 1 and 2% pilocarpine instillation.

We examined ciliary body thickness (CBT) of 11 human eyes after 1% followed by 2% pilocarpine instillation using ultrasound biomicroscopy. The examination revealed that CBT decreased in 2 eyes after 1% pilocarpine treatment but increased after 2% pilocarpine. In the eyes with decreased CBT by 1% pilocarpine, the pretreatment ciliary bodies were thicker and the changing rate of CBT induced by 2% pilocarpine, although not decreased, was also smaller than that in CBT-increased eyes. These findings showed that the ciliary body was under relatively strong stimulus of endogenous acetylcholine and has subsensitivity to both 1 and 2% pilocarpine in CBT-decreased eyes. Configurative changes in CBT-decreased eyes may be explained by the localization of different muscarinic receptor subtypes in different portions of the ciliary muscle.

Adult↗

PubMed2000

Effective treatment with topical cyclosporin A of a patient with Cogan syndrome.

The purpose of this report is to describe the effective treatment of severe anterior segment inflammation due to Cogan syndrome through the use of topical administration of cyclosporin A. A 47-year-old female patient had been experiencing headaches and difficulties with her vision. Subsequent examination revealed the sudden onset of bilateral conjunctival injection and swelling of bilateral auricles. Despite the multiple treatment (systemic and topical corticosteroid and antibiotic therapy), necrotizing scleritis had appeared bilaterally and the scleral wall was thinning. Topical administration of 1% cyclosporin A was applied to both eyes 4 times a day. After 2 months of this therapy, the epithelial tissue covered the necrotizing tissue and her symptom of ocular pain was relieved and her corrected visual acuity was improved. This is the first case exhibiting that topical cyclosporin A is an effective treatment for severe anterior segment inflammation associated with Cogan syndrome.

Administration, Topical↗

PubMed2000

Ceramide-induced cell death in cultured rat retinal pigment epithelial cells.

We investigated whether retinal pigment epithelial (RPE) responds to ceramide, a known second messenger of apoptosis. RPE cells were isolated by 6-8 day old Long Evans rat eye. We used MTS assay for viability test, and used Hoechst 33552 and propidium iodide for apoptotic cell staining. In cultured rat RPE cells, the addition of membrane-permeable ceramide induced apoptosis-like cell death rapidly. RPE cell death was dependent on C2-ceramide concentration. The effective dose (ED50) of C2-ceramide was 23.64 microM. Ceramide-induced RPE cell death was inhibited by zVAD-fmk, a CPP32-like protease inhibitor. Our findings indicated that ceramide in RPE cell death functions upstream of CPP32-like proteases.

Amino Acid Chloromethyl Ketones↗

PubMed2000

Visual processing in patients with macular hole.

Macular hole is a specific disease of the central retina that affects central visual acuity and central visual field. The purpose of this study is to investigate the alteration of visual processing in patients with macular hole who had small central scotoma. Six patients with macular hole participated in this study. We used positron emission tomography (PET) to measure task-related changes in regional cerebral blood flow to identify regions of the brain activated during visual stimulation. Three tasks were performed in each eye: control task, checkerboard task, shape-discrimination task. Checkerboard stimuli caused a greater blood flow activation response in normal eyes than in affected eyes at the occipital cortex. The area involved in the macular hole appeared to be 20 mm or more anteriorly from the occipital pole. The Shape-discrimination task in affected eyes activated angular gyrus, inferoparietal lobule, and middle frontal gyrus. Our findings demonstrated greater confidence in Horton's new retinotopic map than in Holmes' retinotopic map in cortical areas involved in macular function. The dorsal pathway of the visual system was activated more than the ventral pathway in patients with macular hole.

Aged↗

PubMed2000

Time course of the size of the dark area in macular holes by scanning laser ophthalmoscopy.

To investigate the time course of idiopathic macular hole development by scanning laser ophthalmoscopy (SLO), we examined 28 eyes of 25 patients with macular holes. Cases included four eyes with stage 1-B disease (foveal detachment), 3 with stage 2 (break at the fovea), 13 with stage 3 (full-thickness macular hole), and 8 with stage 4 (hole with pseudo-operculum, posterior vitreous detachment), according to Gass's classification. The diameter of the dark area around the macular holes was recorded by SLO using a helium-neon laser. We compared the percent difference of dark area recorded between initial findings and these after three months. The size of the dark area increased after three months in eyes with stages 1, 2, and 3 disease. The size of the dark area in stage 4 disease decreased after 3 months. In the stage before posterior vitreous detachment (PVD), the dark areas increased. Once PVD occurred, these areas decreased. These findings suggested that the time course of the dark area may be related to the development of PVD.

Aged↗

PubMed2000

Müller cells in developing rats with inherited retinal dystrophy.

Morphology and enzymes of Müller cells in the developing retina of RCS (Royal College of Surgeons) rats were investigated. RCS (rdy/rdy) rats with inherited retinal dystrophy were studied and RCS (-/+) rats served as normal controls. Rats underwent intracardiac perfusion with 4% paraformaldehyde and the eyes were enucleated on postnatal days P1, 4,10, 21, 35, and 100. Eyes were then fixed with 4% paraformaldehyde, and silver enhancing technique was applied to show glutamine synthetase (GS) and glial fibrillary acidic protein (GFAP). For solubilized retinas, Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to detect GS and GFAP in the extracts. Immunohistochemistry showed GS expression first on P10. It increased later in both normal and dystrophic retinas. GFAP was not expressed in normal retinas, but Müller cells of dystrophic retinas were stained on P35 and P100. GS immunoblots were recognized on P21 and later in both normal and dystrophic retinas with similar densities, while GFAP immunoblots were observed only on P35 and P100, and only in dystrophic retinas. ELISA demonstrated increased GS concentrations with the development in both normal and dystrophic retinas, but no significant difference was observed between them. GFAP concentrations had no significant difference on P21 between both groups, those of normal ones remained unchanged later, while those of dystrophic rats were remarkably increased on P35 and P100. Müller cells might be affected following the progressive degeneration of photoreceptor cells and react to the glio-neuronal relationship.

Animals↗

PubMed2000

Müller cells in the preconditioned retinal ischemic injury rat.

The role of Müller cells in the preconditioned retinal ischemic injury rat was investigated. In anesthetized Sprague Dawley rats, retinal ischemia for 5 minutes constituted the preconditioning stimulus for the left eye. After 24 hours, both eyes were clamped for 60 minutes. In 30, 60, 90, and 120, minutes and 1 day, 3 days, and 7 days after ischemia, electroretinograms were recorded, and the eyeballs were enucleated. After fixation with 4% paraformaldehyde, the avidin-biotin-peroxidase technique was applied to show glutamine synthetase (GS) and glial fibrillary acidic protein (GFAP). Furthermore, for the solubilized retinas, Western blot analysis and enzyme-linked immunosorbent assay were performed to detect GS and GFAP in the extracts. Preconditioning performed 24 hours before ischemia significantly improved the recovery of the a-, and b-waves 1 day after 60 minute ischemia. In the 30, 60, 90, and 120 minutes after ischemia, the recovery of the a-wave only was observed. There was a nonsignificant trend toward greater recovery in the first 120 minutes after 60 minute ischemia, especially in the b-wave. GS immunoreactivity had no significant difference between non-preconditioned and preconditioned groups 30, 60, 90, and 120 minutes after ischemia. In 1 day after ischemia, GS immunoreactivity decreased in both groups. In 3 and 7 days after ischemia, GS immunoreactivity recovered only in the preconditioned group. The retinas at 3 and 7 days after 1 hour of ischemia showed increased GFAP immunoreactivity in the non-preconditioned group. In the preconditioned group, only slight GFAP immunoreactivity was observed. These results suggested that the mechanism of preconditioned retinal ischemia may be related to Müller cells in the retina.

Animals↗

PubMed2000

Auto iris pigment epithelial cell transplantation in patients with age-related macular degeneration: short-term results.

Autologous iris pigment epithelial cell transplantation was performed on patients with exudative age-related macular degeneration (AMD). Autologous IPE cell culture was performed using autologous serum after iridectomy in 7 patients with AMD. The cell suspensions (2 approximately 20 x 10(4) cells) were transplanted into the submacular lesion of individuals after removal of neovascular membranes. Subsequent ophthalmological examinations, including best corrected visual acuity and fluorescein or indocyanine green angiography, were performed. In addition, 15 patients with AMD, who underwent removal of neovascular membrane without transplantation, were evaluated as non randomized controls. Varying degrees of atrophy or defects of choriocapillaris and retinal pigment epithelium were observed in all of the patients. No cystoid macular edema or fluorescein leakage was observed after treatment, but window defects were present. No patient had decreased visual acuity. One treated patient developed mild subretinal fibrosis and an other patient developed mild preretinal fibrosis, however no difference was significant when compared with the control. In conclusion, the treatment resulted in no significant improvement in macular function, as compared with the control; however, no rejection or deterioration in visual acuity occurred up to the 13 month follow up.

Aged↗

PubMed2000

A new Leu253Arg mutation in the RP2 gene in a Japanese family with X-linked retinitis pigmentosa.

PURPOSE: To identify the clinical findings in a Japanese family with X-linked retinitis pigmentosa associated with mutation in codon 253 (Leu253Arg) in the RP2 gene. METHODS: Case reports included clinical features and results of fluorescein angiography, electroretinogram, kinetic visual field testing, and DNA analysis. Two affected hemizygotes with retinitis pigmentosa associated with transversion mutations in codon 253 (Leu253Arg) of the RP2 gene and the obligate carriers were examined. RESULTS: A novel Leu253Arg mutation of the RP2 gene was found to cosegregate with retinal degeneration in two affected males and two carriers in female heterozygote in a Japanese family. The ophthalmic findings in hemizygote showed severe retinal degeneration. In the obligate carrier, mild chorioretinal degeneration was observed in both eyes but a tapetal-like reflex of the fundus was not apparent. CONCLUSIONS: The mutation at codon 253 of the RP2 gene is the first mutation reported in a Japanese family. It is concluded that the mutation of the RP2 gene also causes the X-linked retinitis pigmentosa in Japanese patients.

Adult↗

PubMed2000

[A case of acute diffuse atrophy of retinal pigment epithelium].

PURPOSE: To report a case of acute bilateral visual disturbance where the ocular fundus changed like retinitis pigmentosa in a short time. PATIENT: 26-year-old man. FINDINGS: In initial examination, the patient's visual acuity was 0.01 OD and 0.02 OS with myopic correction, but his fundus did not look abnormal. Fluorescein angiogram showed marked background hyperfluorescence and dye leaking to the vitreous. After 2-3 weeks, the fundus appearance changed like retinitis pigmentosa. Best corrected visual acuity became 1.0 OD and 0.9 OS after steroid pulse therapy. We were unable to find the cause of this disease in spite of blood tests and other examinations. CONCLUSION: This case with acute diffuse atrophy of retinal pigment epithelium and damage of the function of blood-retinal barrier was considered remarkably rare.

Acute Disease↗

PubMed2000

Autologous iris pigment epithelial cell transplantation in monkey subretinal region.

PURPOSE: To establish autologous iris pigment epithelial (IPE) cell transplantation in monkey eyes. METHODS: Autologous IPE cells from three monkeys were obtained by peripheral iridectomy, and cell culture was performed with autologous serum. The cultured cells were labeled with DiI, and transplantation was performed by transvitreal approach to the submacular region of each monkey. Fundus examination, photography, and fluorescein angiography were performed monthly. Histochemical analysis and electron microscopy were also performed six months after transplantation. RESULTS: Auto IPE cells grew well in auto serum. No fluorescein leakage or retinal thickening was observed after submacular transplantation. We demonstrated the presence of autologous IPE cells in the region 6 months after transplantation by histologic and electron microscopic examination. Electron microscopy also demonstrated that the transplanted cells appeared to be less pigmented and to have less mitochondria than did the host retinal pigment epithelial cells. However, some transplanted cells looked as if they embrace the photoreceptor outer segments. DISCUSSION: Autologous IPE cell culture of the monkey was established using auto serum. Six months after transplantation of the cultured IPE cells, we could still observe the cells in the region. This method may have potential use in human disease that requires autologous cell transplantation to prevent host-graft rejection, and to provide an alternative substance that functions like retinal pigment epithelium.

Animals↗

PubMed2000

[Expression of the varicella zoster virus thymidine kinase and cytokines in patients with acute retinal necrosis syndrome].

PURPOSE: Acute retinal necrosis (ARN) is caused by varicella zoster virus (VZV) infection. In this study, we investigated the activity of this virus and expressions of some cytokines. PATIENTS AND METHODS: The expression of VZV thymidine kinase and some cytokines were investigated by reverse transcriptase-polymerase chain reaction (RT-PCR) in 9 eyes of 8 patients with ARN. RESULTS: Thymidine kinase expression was observed in all samples except one. Several cytokines, such as interferon (IFN) gamma, tumor necrosis factor (TNF) alpha, interleukin (IL)-1 beta, IL-6, and transforming growth factor (TGF) beta 1 were observed in the samples. Among these cytokines, a statistically significant expression of IFN gamma was observed in the samples of ARN, when compared to those of proliferative vitreoretinopathy (PVR) or other uveitis. The expression of IFN gamma also decreased during successive follow-ups. CONCLUSION: These cytokines may play an important role in the immune response in ARN.

Adolescent↗

PubMed2000

A frequent 1085delC/insGAAG mutation in the RDH5 gene in Japanese patients with fundus albipunctatus.

PURPOSE: To identify the frequency of a mutation of the RDH5 gene in Japanese patients with hereditary retinal degeneration and to characterize clinical findings for the patients associated with a 1085delC/insGAAG mutation in the RDH5 gene. METHODS: Mutation screening by single-strand conformation polymorphism was performed on 6 patients with fundus albipunctatus and 150 patients with autosomal recessive retinitis pigmentosa. The DNA fragment that showed abnormal mobility on SSCP was then sequenced. Clinical features were characterized by visual acuity, slit-lamp biomicroscopy, electroretinography, fluorescein angiography, kinetic visual field testing, and dark adaptometry. RESULTS: A novel 1085delC/insGAAG mutation in the RDH5 gene was identified in all 6 patients, from 4 unrelated families with fundus albipunctatus. The ophthalmic findings of each affected member were very similar, which may provide the natural course of the phenotype produced by the 1085delC/insGAAG mutation. CONCLUSIONS: A homozygous1085delC/insGAAG mutation in the RDH5 gene produces fundus albipunctatus in Japanese patients. These findings suggest that this mutation was a founder effect in Japanese patients with fundus albipunctatus.

Adult↗

PubMed2000

Adrenomedullin in cultured human retinal pigment epithelial cells.

PURPOSE: To determine whether adrenomedullin (ADM), a vasorelaxant peptide is produced and secreted by human retinal pigment epithelial (RPE) cells, whether ADM expression is regulated by inflammatory cytokines and a growth factor, and whether ADM has proliferative effects on these cells. METHODS: Production and secretion of ADM by cultured human RPE cells were examined by Northern blot analysis and radioimmunoassay. Regulation of the ADM expression by basic fibroblast growth factor, interferon (IFN)-gamma, tumor necrosis factor-alpha, interleukin (IL)1beta, or all-trans-retinoic acid was studied. In addition, proliferative effects of ADM on human RPE cells were examined by modified 3-(4,5-dimetylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: ADM mRNA was expressed constitutively in all three human RPE cell lines (F-0202, D407, and ARPE-19) examined. Immunoreactive ADM was detected in the cultured media by radioimmunoassay. Sephadex G-50 column chromatography of the cultured medium showed a single peak eluting in the position of ADM-(1-52). Treatment with IFN-gamma or IL-beta increased ADM mRNA levels and immunoreactive-ADM levels in the medium in dose- and time-dependent manners in ARPE-19 cells. Exogenously added ADM increased the number of F-0202 cells and ARPE-19 cells, and the treatment with ADM antibody or ADM-(22-52) (an ADM antagonist) decreased it. CONCLUSIONS: Human RPE cells produced and secreted ADM. IFN-gamma and IL-1beta induced ADM expression in ARPE-19 cells. Furthermore, ADM stimulated proliferation of RPE cells. These results raise the possibility that ADM is related to the pathophysiology of some inflammatory and proliferative ocular diseases.

Adrenomedullin↗

PubMed1999

Possible mechanism for the decrease of mitochondrial aspartate aminotransferase activity in ischemic and hypoxic rat retinas.

Glutamate is believed to be an excitatory amino acid neurotransmitter in the retina. Enzymes for glutamate metabolism, such as glutamate dehydrogenase, ornithine aminotransferase, glutaminase, and aspartate aminotransferase (AAT), exist mainly in the mitochondria. The abnormal increase of intracellular calcium ions in ischemic retinal cells may cause an influx of calcium ions into the mitochondria, subsequently affecting various mitochondrial enzyme activities through the activity of mitochondrial calpain. As AAT has the highest level of activity among enzymes involved in glutamate metabolism, we investigated the change of AAT activity in ischemic and hypoxic rat retinas and the protection against such activity by calpain inhibitors. We used normal RCS (rdy+/rdy+) rats. For the in vivo studies, we clamped the optic nerve of anesthetized rats to induce ischemia. In the in vitro studies, the eye cups were incubated with Locke's solution saturated with 95% N2/5% CO2. The activity of cytosolic AAT (cAAT) was about 20% of total activity, whereas mitochondrial AAT (mAAT) was about 75% in rat retina. Ninety minutes of ischemia or hypoxia caused a 20% decrease in mAAT activity, whereas cAAT activity remained unchanged. To examine the contribution of intracellular calcium ions to the degradation of mAAT, we used Ca2+-free Locke's solution containing 1 mM EGTA, ryanodine (Ca2+ channel blocker), and thapsigargin (Ca2+-ATPase inhibitor). In the present study, thapsigargin in Ca2+-free Locke's solution, but not ryanodine in this solution, was found to prevent AAT degradation. AAT degradation was also prevented by calpain inhibitors (Ca2+-dependent protease inhibitor) such as calpeptin at 1 nM, 10 nM, 0.1 microM, 1 microM and 10 microM, and by calpain inhibitor peptide, but not by other protease inhibitors (10 microM leupeptin, pepstatin, chymostatin). Additionally, we determined the subcellular localization of calpain activity and examined the change of calpain activity in ischemic rat retinas. Our results suggest that decreased activity of mAAT in ischemic and hypoxic rat retinas might be evoked by the degradation by calpain-catalyzed proteolysis in mitochondria.

Animals↗

PubMed1999

The genomic organization of type I keratin genes in mice.

We isolated two new keratin cDNAs by screening a cDNA library constructed from poly(A)+ RNA of the dorsal and abdominal skin of C57BL/10J mice with a probe of human KRT14. Due to its high sequence homology to human keratin 17 cDNA, one full-length cDNA is most likely to be mouse keratin 17 (Krt1-17) cDNA. The other is the putative full-length cDNA of a novel type I keratin gene, designated Krt1-c29. These two keratin genes were mapped to the distal portion of Chromosome 11, where the mouse keratin gene complex-1 (Krt1) is localized. To elucidate the genomic organization of Krt1 in mice, we carried out genetic and physical analyses of Krt1. A large-scale linkage analysis using intersubspecific backcrosses suggested that there are two major clusters in Krt1, one containing Krt1-c29, Krt1-10, and Krt1-12 and the other containing Krt1-14, -15, -17, and -19. Truncation experiments with two yeast artificial chromosome clones containing the two clusters above have revealed that the gene order of Krt1 is centromere-Krt1-c29-Krt1-10-Krt1-12-Krt1-13-K rt1-15-Krt1-19-Krt1-14-K rt1-17-telomere. Finally, we analyzed sequence divergence between the genes belonging to the Krt1 complex. The results clearly indicated that genes are classified into two major groups with respect to phylogenetic relationship. Each group consists of the respective gene cluster demonstrated by genetic and physical analyses in this study, suggesting that the physical organization of the Krt1 complex reflects the evolutionary process of gene duplication of this complex.

Amino Acid Sequence↗