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K Shinoda

Publications and source records attributed to K Shinoda.

At least 37 records · Page 2Linked to original sources

A melanosome-associated monoclonal antibody J1 recognizes luminal membrane of prelysosomes common to biogenesis of melanosomes and lysosomes.

Melanogenesis cascade may be directly or indirectly linked to the dynamics of endosome-lysosome biogenesis. This study aims to identify how and to what extent the endosome-lysosome system is involved in melanosome biogenesis, by utilizing a novel melanogenesis marker, J1, which we identified in the process of developing monoclonal antibodies (MoAbs) against human melanosomes. The antigenic epitope of MoAb J1 was expressed by all of the melanotic and nonmelanotic cells examined. It was expressed primarily by granular structures located in regions proximal to the Golgi complex. Most of MoAb J1 positive granules were co-stained with melanogenic markers, tyrosinase or tyrosinase-related protein (TRP-1). The epitope of MoAb J1 was also coexpressed by most, but not all, of LGP85 (a lysosomal marker) positive granules in both melanoma and non-melanoma cells, indicating that MoAb J1 recognizes a subset of lysosomal vesicles. MoAb J1 did not, however, react with vesicles with late/early (syntaxin 8/ EEA1) endosomal markers. Further examination using fluorophore-labeled pepstatin, a marker of lysosomal luminal content, confirmed that MoAb J1 specifically recognizes the luminal surface of lysosomes. These results indicate that MoAb J1 possesses an antigen epitope that is expressed in the luminal component of prelysosomal granules which are involved in the biogenesis cascade common to both melanosomes and lysosomes. We suggest that tyrosinase family protein, tyrosinase and TRP-1 are transported to melanosomes from TGN via these prelysosomal granules after being transiently transported to late endosomes.

Animals↗

Clinical application of a new compact computed tomography system for evaluating the outcome of regenerative therapy: a case report.

BACKGROUND: The limitations of conventional 2-dimensional (2-D) radiographs have been emphasized in many studies. Because the image is a 2-D map of the 3-dimensional (3-D) tooth and periodontal tissue, these structures may be superimposed on each other. Recently, 3-D image analysis by computed tomography (CT) was introduced to dentistry, but it has been used only rarely in the periodontal field. METHODS: The purpose of this case report was to introduce the clinical application of a newly developed, compact computed tomography system for evaluating the outcome of regenerative therapy in an adult patient with periodontitis (male, aged 55 years). To evaluate the bone defects including furcation involvement, we took an intraoral radiograph and performed CT scanning on tooth #19. We recorded a number of clinical parameters at the time of surgery. We compared the images of the bone defect obtained by CT images and the conventional radiograph with the true bone defect morphology at the surgical site. RESULTS: One year after the periodontal surgery, we again took CT scans and a conventional radiograph, and the new images of the bone defect were compared and evaluated. The new CT images were reconstructed for 3-D evaluation and compared with those taken before surgery. The CT provides 3-D images of excellent quality for evaluating the morphology of the periodontal bone defect and furcation involvement. CONCLUSIONS: We consider that CT is a useful aid in periodontal examination and diagnosis, and in the prediction and evaluation of periodontal treatment outcomes.

Alveolar Bone Loss↗

[Objective evaluation of visual field loss in a patient with branch retinal artery occlusion and brain infarction].

PURPOSE: To evaluate visual field loss using multifocal ERG(m-ERG), multifocal VEP(m-VEP), and Heidelberg Retina Flowmeter(HRF) in a patient with branch retinal artery occlusion(BRAO) and brain infarction. CASE: A 38-year-old man noticed inferior-nasal visual field loss in the left eye, and was referred to Keio University Hospital. He suffered from paralysis in the left leg due to brain infarction at the age of 24. However, he had not noticed visual field loss due to the brain infarction. His left fundus showed retinal edema in the area of a superior-temporal retinal artery occulusion. He was diagnosed as having BRAO. The Goldmann and Humphry perimetric examinations revealed homonymous quadrantanopia in the upper left field as well as inferior visual field defect in the left eye. RESULTS: Both m-ERG and m-VEP, especially second-kernel responses, were reduced in the affected retinal area of BRAO. But only m-VEP was affected in the corresponding area of homonymous quadrantanopia in the upper left field. The retinal flow in the area with BRAO evaluated by HRF was decreased in some areas and not in others, suggesting that retinal function was not necessarily consistent with retinal circulation. CONCLUSIONS: m-ERG and m-VEP are useful To differentiate retinal lesions from brain lesions in visual field loss.

Adult↗

Systematic synthesis of sulfated sialyl-alpha-(2 --> 3)-neolactotetraose derivatives and their acceptor specificity for an alpha-(1 --> 3)-fucosyltransferase (Fuc-TVII) involved in the biosynthesis of L-selectin ligand.

Sulfated sialyl-alpha-(2 --> 3)-neolactotetraose (IV3NeuAcnLcOse4) derivatives at C-6 of GlcNAc (6-O-sulfo), terminal Gal (6'-O-sulfo), and both GlcNAc and Gal (6,6'-di-O-sulfo) residues have systematically been synthesized. (Methyl 5-acetamido-4,7,8,9- tetra-O-acetyl-3,5-dideoxy-D-glycero-alpha-D-galacto-2-nonulopyranosy lonate)-(2 --> 3)-2,4-di-O-benzoyl-6-O-levulinoyl-D-galactopyranosyl trichloroacetimidate was coupled with 2-(trimethylsilyl)ethyl (2-acetamido-2-deoxy- 3-O-benzyl-6-O-p-methoxyphenyl-beta-D-glucopyranosyl)-(1 --> 3)-(2,4,6-tri-O-benzyl-beta-D-galactopyranosyl)-(1 --> 4)-2,3,6-tri-O-benzyl-beta-D-glucopyranoside to give the suitably protected pentasaccharide which, upon selective removal of the p-methoxyphenyl and/or levulinoyl groups at C-6 of the GlcNAc and the terminal Gal residues, successive O-sulfation(s) and deprotection, afforded the desired three sulfated IV3NeuAcnLcOse4 derivatives. Acceptor specificity of the synthetic IV3NeuAcnLcOse4 probes for a human alpha-(1 --> 3)-fucosyltransferase (Fuc-TVII) was examined to study the biosynthetic pathway of L-selectin ligand. Only the 6-sulfated derivative at C-6 of GlcNAc was recognized by Fuc-TVII to give 6-O-sulfo sialyl LeX.

Binding, Competitive↗

Induction and inhibition of testicular germ cell apoptosis by fluoroacetate in rats.

Fluoroacetate (FA), an inhibitor of aconitase, is known to lower the intracellular level of adenosine triphosphate (ATP), which recently has been suggested to be a possible determinant of the form of cell death, apoptosis or necrosis. To investigate which form of germ cell death occurs in FA-induced testicular toxicity, adult Sprague Dawley rats were given a single oral dose of FA (0.5 or 1.0 mg/kg) and euthanized at 3, 6, 12, 24, 48, and 72 h thereafter. Germ cell degeneration was histologically first found in early round spermatids at stage I and in spermatogonia at stages II-IV of seminiferous tubules 6 and 12 h, respectively, after dosing. Degenerating spermatogonia exhibited characteristic features of apoptosis as demonstrated by both electron microscopy and in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), whereas spermatids did not. At the 24 and 48 h time points, degenerating spermatids were continually present and subsequently formed multinucleated giant cells, while the number of degenerating spermatogonia and TUNEL-labeled spermatogonia was drastically and/or significantly decreased compared to those from the control group, indicating that spontaneous male germ cell apoptosis is inhibited. Coincident with these morphological changes, DNA laddering on gel electrophoresis was apparent only 12 h after dosing. The results demonstrate that FA induces either apoptosis or necrosis of male germ cells in the early stage after dosing and subsequently inhibits spontaneous apoptosis.

Aconitate Hydratase↗

Changes in serum alpha2u-globulin levels in male rats given diethylstilbestrol and applicability to a screening test for endocrine-disrupting chemicals.

alpha2u-Globulin (AUG) is a major rat urinary protein, which has a molecular weight of 16 kDa (kidney type) or 19 kDa (native type). The biosynthesis of this protein is under multi-hormonal regulation. In this study, we investigated changes in serum AUG level and their association with changes in the reproductive organs of male rats after the administration of the estrogenic chemical, diethylstilbestrol (DES) at doses ranging from 0.01 mg/kg per day to 100 mg/kg per day by gavage for 14 days. Our aim was to establish basic data for the development of a new screening method for endocrine disrupting chemicals based on serum AUG levels. DES treatment decreased the weight of testes in a dose-dependent manner; and was accompanied by atrophic histopathological changes in testes. Testis weights were significantly decreased by the group given 1 mg/kg per day DES; however, histopathological abnormalities were found in the group given 0.1 mg/kg per day DES. In four of five animals in the group given 1 mg/kg per day there was no significant decrease in testis weight and only a slight or moderate degeneration of the pachytene spermatocytes. Despite these findings, serum AUG levels in this group decreased markedly, while the serum AUG level markedly decreased even in the animals with no histopathological change in the 1 mg/kg per day or 0.1 mg/kg per day groups with no histopathological change also showed decreased serum AUG level. These results suggest that the serum AUG level may be a sensitive parameter for detecting the activity of estrogenic chemicals in intact male rats. Although a uterotropic assay has been proposed for immature female or ovariectomized female rats and is currently undergoing validation studies internationally, there is no screening method for estrogenic chemicals in intact male animals. More data on AUG changes by treatment with other estrogenic chemicals are needed in order to determine the sensitivity and specificity of this response to estrogens. Nonetheless, an AUG-based screening test for estrogenic chemicals may be useful owing to its applicability to conventional toxicity studies and an apparently higher sensitivity of this parameter compared to organ weight change or histology of testis in intact male rats and applicability to conventional toxicity studies.

Alpha-Globulins↗

A new method for quantification of metamorphopsia in patients with epiretinal membrane

Purpose: We have developed a new method for quantification of metamorphopsia and applied it to study distorted vision resulting from epiretinal membrane (ERM).Patients and Methods: We prepared a modified Amsler chart, which was a square grid formed by black lines on a white background with 12 cm on a side and divided into 2-cm quadrants. The patients were asked to trace any straight lines on the chart which appeared irregular or curved. The length of all lines traced by the patients was measured except for the outer frame. The total length of the chart itself was 1.200 mm, but it would appear longer in patients with metamorphopsia. In addition, the severity of metamorphopsia was scored subjectively as follows: 1, absent; 2, slight; 3, mild; 4, moderate; and 5, severe. The relationship of the length to the score and to the visual acuity were analysed. Sixty-three patients with unilateral ERM were examined.Results: The length of the line ranged from 1,200 to 1,259 mm (mean 1,223.3 mm) and was correlated significantly to the score. Conclusions: This method might be applied usefully in evaluating the severity of metamorphopsia and the surgical outcome of eyes with ERM.

Journal Article↗

Intracellular vesicular trafficking of tyrosinase gene family protein in eu- and pheomelanosome biogenesis.

The intracellular vesicular trafficking in the melanosome biogenesis (melanogenesis) is reviewed with the incorporation of our own experimental findings. The melanosome biogenesis involves four stages of melanosome maturation, which reflect the transport of structural and enzymatic proteins from Golgi (trans-Golgi network: TGN) to the melanosomal compartment and their organization therein. The major melanosomal proteins include tyrosinase gene family protein (tyrosinase and tyrosinase-related protein; TRP), lysosome-associated membrane protein (Lamp) and gp100 (pmel 17). They are glycosylated in the endoplasmic reticulum, and transported by vesicles from the TGN to the melanosomal compartment. During the formation of transport vesicles, they assemble on the cytoplasmic face of the TGN to select cargo by interacting directly or indirectly with coat proteins. Tyrosinase and TRP-1 possess the dileucine motifs at the cytoplasmic domain, to which adapter protein-3 binds to transport them from the TGN to stage I melanosomes (related to late endosomes) and then to stage II melanosomes. A number of small guanosine triphosphate-binding proteins, including rab 7, appear to be involved in this vesicular transport. Phosphatidyl inositol 3 kinase also regulates this membrane trafficking of melanosomal glycoprotein. Eumelanogenesis is controlled by melanocyte-stimulating hormone, and all three tyrosinase gene family proteins are transported from the TGN to stage II melanosomes that are elliposoidal and contain the structural matrix of filaments/lamellae. In contrast, pheomelanogenesis is primarily regulated by agouti signal protein, and only tyrosinase is transported from stage I melanosomes to stage II melanosomes that are spherical and related to lysosomes. Because of the absence of TRP-1 and TRP-2 in pheomelanogenesis, it may be suggested that tyrosinase is involved in lysosomal degradation after forming dopaquinone, to which the cysteine present in the lysosomal granule binds to form cysteinyldopas that will then be auto-oxidized to become pheomelanin.

Animals↗

Ultrasound biomicroscopy for examination of the sclerotomy site in eyes with proliferative diabetic retinopathy after vitrectomy.

PURPOSE: We evaluated the capability of ultrasound biomicroscopy (UBM) to predict fibrovascular proliferation at sclerotomy sites in eyes with postoperative vitreous hemorrhage due to proliferative diabetic retinopathy (PDR). METHODS: Ultrasound biomicroscopy was used for examining the sclerotomy sites in 13 eyes of 11 patients with PDR experiencing postoperative vitreous hemorrhage (PDR group). Thirty-nine sclerotomy sites (all entry sites of each eye) were examined before reoperation, and the UBM images were compared with findings obtained during revision of the vitrectomy. Thirteen eyes of 13 patients undergoing vitrectomy for nondiabetic diseases were used as controls and examined after vitrectomy. RESULTS: The UBM images were classified into the following four categories: A, tent; B, spheroid; C, trapezoid; and N, none. The findings were distributed as follows in the PDR group: category A, 18%; B, 5%; C, 56%; and N, 21 %; and as follows in the control group: category A, 28%; B, 5%; C, 5%; and N, 62%. In the PDR group, 11 of 12 sclerotomy sites disclosing fibrovascular proliferation possessed the trapezoidal image. Mean length of trapezoidal base was 2.49+/-0.97 mm and 1.51+/-0.75 mm in the groups with and without fibrovascular proliferation, respectively (P<0.01). The average relative reflectivity of the trapezoidal image against the sclera was 0.501+/-0.169 in the fibrovascular proliferation group and 0.891+/-0.183 in the fibrous ingrowth group (P<0.01). CONCLUSION: Ultrasound biomicroscopy is useful in detecting fibrovascular proliferation at sclerotomy sites because a large and low-reflecting trapezoidal UBM image is highly correlated to its presence.

Adult↗

Ultrastructural and immunohistochemical findings in five patients with vitreomacular traction syndrome.

OBJECTIVE: To evaluate pathologic features of vitreomacular traction syndrome (VMT). METHODS: Preretinal membranes removed from five patients during vitreous surgery for VMT syndrome were evaluated by electron microscopy (n = 4) and immunohistochemistry (n = 1). RESULTS: Electron microscopic examination revealed large segments of internal limiting lamina (ILL) in three of the four cases. Other extracellular features included two types of abnormal collagen fibrils, determined to be type I and fibrous long-spacing collagen. The myofibrocyte was the predominant cell type in all cases. Collagen types I, II, and III, as well as glial fibrillary acidic protein and vimentin, were identified by immunohistochemistry. CONCLUSIONS: Epiretinal membranes in eyes with VMT syndrome adhered tightly to the ILL with abnormal collagen and contractile elements that included myofibrocytes.

Aged↗

Melanocytoma of the left optic nerve head and right retrobulbar optic neuropathy compressed by a tuberculum sellae meningioma.

A 40-year-old woman had a highly pigmented, slightly elevated tumor on the left optic disc. She had no visual disturbance in the left eye. The tumor was stationary for 5 years. At age 45 years, she complained of decreased visual acuity in the right eye. Magnetic resonance imaging showed a right-shifted homogeneous lesion at the tuberculum sellae. Histopathologic study of the excised lesion revealed interlacing bundles of spindle-shaped fibroblast-like cells with whorl formation. We believe that a relationship between melanocytoma of the optic nerve head and a tuberculum sellae meningioma may exist rather than a chance occurrence, as previously suggested by others.

Adult↗

Infant leukemia suggestive of natural killer cell precursor origin followed an unusual clinical course.

We report a patient with infant leukemia showing the rare phenotypes of CD2+, CD4+, CD7+, CD56+, CD3-, CD5- who followed an unusual clinical course. The patient was a 3-month-old girl who was admitted because of unusual purpura looking like a black ring. On admission WBC count was 85.8 x 10(9)/l and bone marrow aspiration revealed a nucleated cell count of 112.0 x 10(9)/l with 70.8% atypical lymphocytes. On the 3rd hospital day, the WBC count decreased by about 1/5 without chemotherapy and partial remission was obtained. But about 3 weeks later, the WBC count increased again and she died. Based on surface marker analysis, genotypic analysis and autopsy, we diagnosed infant leukemia suggestive of natural killer (NK) cell precursor origin.

Antigens, CD↗

Cytotoxic alkaloids of Pachysandra terminalis.

Four steroidal alkaloids, epipachysamines B (1) and E (2), pachystermine A (3) and pachysamine E (4), were isolated as cytotoxic principles from the MeOH extract of the stems of Pachysandra terminalis SIEB. et ZUCC. (Buxaceae). These alkaloids showed cytotoxic activity against P388 and P388/ADR leukemia cells in vitro. Three of the alkaloids (1-3) were previously isolated from this plant material, and this is the first report of their cytotoxic activity. Pachysamine E (4) is a new alkaloid.

Alkaloids↗

Diagnostic efficacy of three-dimensional images by helical CT for lesions in the maxillofacial region.

The purpose of this study was to evaluate the efficacy of three-dimensional (3-D) images produced with a helical CT for the diagnosis of lesions occurring in the maxillofacial region. Thirty-four patients, who had lesions in the maxillofacial region, were examined by plain radiography (intra and extraoral) and the helical CT. Further, 3-D images were reconstructed from the data provided by the helical CT using the volume rendering method. These images were compared with plain radiographic images and conventional two-dimensional (2-D) CT images in terms of the information they provided for diagnosis. Using the 3-D images for tumors, bone destruction, inner components, extent of the lesion,the relationship between the lesion and surrounding anatomical landmarks, and the roots of the adjacent teeth were observed in overall views. We conclude that 3-D images produced by helical CT may provide useful information for the diagnosis of lesions.

Adult↗

[Comparison of scleral buckling and vitrectomy for superior retinal detachment caused by flap tears].

PURPOSE: We compared the surgical results of vitrectomy and scleral buckling for uncomplicated superior retinal detachment caused by flap tears. SUBJECTS AND METHODS: Included in the study were 225 phakic eyes of 225 patients undergoing primary surgery from January 1990 to December of 1996 for superior retinal detachment caused by flap tears; all eyes had been followed for longer than six months after surgery. The choice of procedures was based on each surgeon's preference. The cases were evaluated retrospectively and the surgical outcome and the rate of complications compared between the two groups of eyes. RESULTS: Initial and final anatomical success rate were 92% and 100% after each procedure. Redetachment after the first procedure was due to new retinal breaks in 5 eyes, reopening of original breaks in 2 eyes of vitrectomy cases, and malpositioned buckle in 11 eyes of scleral buckling cases. Proliferative vitreoretinopathy occurred in 3 eyes of vitrectomy cases. CONCLUSION: Primary vitrectomy was as successful as scleral buckling for superior rhegmatogenous retinal detachment. Although a high incidence of postoperative cataract formation was a major drawback, vitrectomy had some advantages over to scleral buckling.

Adult↗

Macular hole retinal detachment in highly myopic eyes: ultrastructure of surgically removed epiretinal membrane and clinicopathologic correlation.

PURPOSE: To elucidate the pathogenesis of macular hole retinal detachment (RD) in highly myopic eyes by investigating the ultrastructure of surgically removed epiretinal membranes (ERM). METHODS: Five consecutive Japanese patients with macular hole RD in highly myopic eyes underwent vitrectomy with attempted removal of the ERM around the hole. The surgical specimens were examined by light microscopy and by scanning and transmission electron microscopy. RESULTS: Extremely thin, translucent sheets of epiretinal tissue were harvested from all patients during surgery, resulting in successful retinal reattachment. Ultrastructural examination revealed that the ERM consisted of cortical vitreous and various cellular components. Fibrous astrocytes were the major cell population and extended cytoplasmic processes with membrane-associated vesicles onto the cortical vitreous. Gap junctions were observed between the interdigitating processes of fibrous astrocytes. The cortical vitreous contained abundant newly formed collagen, including fibrous long-spacing collagen, surrounded by sparsely distributed native vitreous collagen. CONCLUSIONS: Active synthesis of new collagen may be regulated by fibrous astrocytes by means of transmission of metabolic substances through gap junctions and cytoplasmic vesicles. The frequent occurrence of newly formed collagen aggregates may subsequently lead to a diffusely condensed posterior cortical vitreous that exerts tangential traction on the posterior retina, causing macular hole RD.

Aged↗

Coexpression of VEGF receptors VEGF-R2 and neuropilin-1 in proliferative diabetic retinopathy.

PURPOSE: To elucidate vascular endothelial growth factor (VEGF)-mediated pathogenesis of fibrovascular proliferation in diabetic retinopathy. METHODS: Fibrovascular tissues were obtained at vitrectomy from 22 cases with proliferative diabetic retinopathy. The half-divided tissues were processed for reverse transcription-polymerase chain reaction (RT-PCR) analysis to examine the expression of VEGF isoforms and their receptors. Paraffin sections of the other half were used for immunohistochemistry for CD34, glial fibrillary acidic protein and VEGF, and in situ hybridization for VEGF. RESULTS: RT-PCR analysis demonstrated the expression of VEGF receptors VEGF-R1, VEGF-R2, and neuropilin-1 in 12, 14, and 14 of 22 cases, respectively. Notably, VEGF-R2 and neuropilin-1 were simultaneously expressed in the identical 14 tissues. The isoform VEGF121 was constitutively expressed in all the tissues examined, whereas the expression of VEGF165 was confined to the 7 tissues that also expressed VEGF-R2 and neuropilin-1. The vascular density of fibrovascular tissues evaluated by immunohistochemistry for CD34 was significantly higher in the cases with the expression of VEGF-R2 and neuropilin-1 than in those without their expression (P < 0.01), whereas VEGF-R1 expression had no such relationship with the vascular density. The fibrovascular tissues that expressed VEGF165 together with VEGF-R2 and neuropilin-1 were found in significantly younger patients (P < 0.01). In situ hybridization and immunohistochemical studies demonstrated that glial cells in the fibrovascular tissues express and produce VEGF. CONCLUSIONS: Coexpression of VEGF-R2 and neuropilin-1 is suggested to facilitate fibrovascular proliferation in diabetic retinopathy.

Adult↗