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Biomedical subjects

Y Obara

Publications and source records attributed to Y Obara.

At least 55 records · Page 3Linked to original sources

Histopathological Features of the Trabecular Meshwork in a Case of Presumed Bilateral Chandler Syndrome.

Background: We describe a patient who had no-table decrease in the number of corneal endothelial cells in both eyes and developed open angle glaucoma without evident iris atrophy and peripheral anterior synechia.Case: A 74-year-old woman. The trabeculectomized angle tissue and iris tissue of her left eye were observed under light and transmission electron microscopy. From Schwalbe's line to the anterior chamber angle, degenerated endothelium-like cells were observed and overgrowth of layers of collagen fiber with varied cycles and basement membrane-like material were noticed. In addition, the morphology of intra-trabecular space was atrophic and occlusive with marked degeneration and exfoliation of endothelial cells in the trabecular space. On the side of the anterior chamber, degenerated endothelium-like cells were observed, the morphology of which was considered to result from abnormal metabolic function in corneal endothelium. Overgrowth of basement membrane-like material onto iridic tissue was not observed.Conclusion: Although slit lamp examination revealed no obvious abnormality, the result of histological examination suggested that this was a case of Chandler's syndrome in a broad sense or was in the early stage of this disease. We also discussed differential diagnosis from other diseases.

Journal Article↗

Differentiation of rat pheochromocytoma cells by fomitellic acids, specific DNA polymerase inhibitors.

Fomitellic acid (FA) A and B are specific inhibitors of DNA polymerase alpha and beta. They showed cytotoxicity against rat pheochromocytoma cells (PC-12 cells) in a concentration-dependent manner. However, after PC-12 cells were cultivated with low concentrations of FAs, the cells extended neurites in greater degree similar to the cells cultivated with nerve growth factor. Another DNA polymerase alpha inhibitor, aphidicolin, also induced neurite outgrowth. Furthermore, PC-12 cells were strongly immunostained with anti-alpha-tubulin or anti-tau antibody after the treatment with FAs. These results suggest that weak inhibition of DNA polymerase activity induces the neurite outgrowth in PC-12 cells.

Animals↗

Changes in cytoskeletal proteins in childhood cataract lenses

Purpose: Approximately 50% of congenital and childhood cataracts seen in the clinic is of undetermined origin. Biochemical analysis of the cataracts is rare. This study analyzes lens proteins to determine the mechanism of congenital and childhood cataracts. Method: We analyzed the lens proteins from 10 young patients after cataract operations, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), densitometry analysis, and Western immunoblotting.Results: Densitometry of separated proteins showed a decrease in the high molecular protein bands of posterior subcapsular cataract (PSC). Specifically, spectrin (235 kDa), filensin (100 kDa), and vimentin (57 kDa) were absent from the SDS-PAGE of PSC. Increases in filensin and vimentin were observed in a Christmas tree cataract and lamellar cataracts. Western immunoblots confirmed the densitometry of SDS-PAGE.Conclusion: These results suggest that changes in cytoskeletal proteins may contribute to congenital and childhood cataracts.

Journal Article↗

[Histopathological features of the trabecular meshwork in a case of presumed bilateral Chandler syndrome].

BACKGROUND: We describe a patient who had notable decrease in the number of corneal endothelial cells in both eyes and developed open angle glaucoma without evident iris atrophy and peripheral anterior synechia. CASE: A 74-year-old woman. The trabeculectomized angle tissue and iris tissue of her left eye were observed under light and transmission electron microscopy. From Schwalbe's line to the anterior chamber angle, degenerated endothelium-like cells were observed and overgrowth of layers of collagen fiber with varied cycles and basement membrane-like material were noticed. In addition, the morphology of intra-trabecular space was atrophic and occlusive with marked degeneration and exfoliation of endothelial cells in the trabecular space. On the side of the anterior chamber, degenerated endothelium-like cells were observed, the morphology of which was considered to result from abnormal metabolic function in corneal endothelium. Overgrowth of basement membrane-like material onto iridic tissue was not observed. CONCLUSION: Although slit lamp examination revealed no obvious abnormality, the result of histological examination suggested that this was a case of Chandler's syndrome in a broad sense or was in the early stage of this disease. We also discussed differential diagnosis from other diseases.

Aged↗

[Analysis of cytoskeletal proteins in rat selenium cataract using two-dimensional electrophoresis].

BACKGROUND AND PURPOSE: Previous studies used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to characterize the modifications to lens proteins during opacification in the selenite model for cataract. To analyze the protein modifications in more detail, two dimensional electrophoresis (2 DE), which is more sensitive than 1 D electrophoresis, was used. RESULTS: The results of 2 DE demonstrated rapid changes in cytoskeletal proteins including spectrin and vimentin at the earliest stages of opacification. CONCLUSION: The results suggested that the mechanism of opacification might involve changes in cytoskeletal proteins during the earliest stages of cataract formation.

Animals↗

[Analysis of proteins during recovery from lens opacity--analysis of selenite cataract model using Sprague-Dawley and Wistar rat].

BACKGROUND AND PURPOSE: The cataract in Sprague-Dawley rats injected with selenite is a dense nuclear opacity that appears by 4 or 5 days after selenite injection and becomes irreversible by 7 days. Injection of Wistar rats with selenite resulted in a similar nuclear opacity by 4 or 5 days that began to recover transparency by 7 days. In this report, the cytoplasmic proteins were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in lenses from Sprague-Dawley and Wistar rats at 4 and 7 days after injection. RESULTS: In the opaque lens cells, degradation of the 31 kDa protein and cytoskeletal proteins (vimentin, spectrin, and actin) was observed during cataract development using SDS-PAGE and western blot analysis. During recovery from opacity, the decreased 31 kDa protein and the vimentin increased. CONCLUSION: The results suggest that the 31 kDa protein and the vimentin may be important for recovery of transparency in a reversible model of cataract formation.

Animals↗

EDTA: a promoter of proliferation in human corneal endothelium.

PURPOSE: To determine whether it is possible to induce proliferation in the endothelium of older donor corneas by treatment of the intact monolayer with EDTA. METHODS: Corneas from donors 52 to 75 years of age were obtained from an eye bank and were usually cut in quarters to increase sample size. The effect of EDTA dose (0.02-2.0 mg/ml) and incubation time (6, 30, and 60 minutes) on endothelial cell-cell contacts was evaluated by staining for ZO-1, a cell junction marker. Cell death was tested by a commercial live-dead assay. Corneal pieces were incubated for 0, 24, 48, or 60 hours in culture medium (M-199, 10% fetal bovine serum, 10 ng/ml epidermal growth factor, 20 ng/ml fibroblast growth factor) before EDTA treatment. After treatment, pieces were incubated in the same medium for 24, 48, 72, or 96 hours to permit cell cycle entry. Tissue was fixed, stained for Ki67 (a marker for late G1-phase through the M-phase), and mounted in medium containing propidium iodide to visualize all nuclei. Confocal images were evaluated by computer (Image software; NIH, Bethesda, MD) to count Ki67-positive and propidium iodide-stained cells. RESULTS: EDTA released corneal endothelial cell-cell contacts in a dose- and time-dependent manner. At doses and incubation times tested, EDTA did not induce significant cell death. Preincubation in culture medium for 24 hours was needed for endothelial cells to efficiently initiate proliferation in response to EDTA. The endothelium of corneas incubated in mitogen-containing medium for up to 108 hours without EDTA treatment did not stain for Ki67. EDTA at 2.0 mg/ml for 60 minutes appeared optimal and stimulated 16% to 18% of the cells to proliferate. Ki67-positive mitotic figures were visible 48 hours after exposure to EDTA. Formation of daughter cells was visible after double-staining for Ki67 and ZO-1. CONCLUSIONS: EDTA released cells from contact inhibition and promoted proliferation in corneal endothelium from older donors. The authors hypothesize that corneal endothelium from older individuals divide in situ when exposed to positive growth factors under conditions in which cells have been transiently released from contact inhibition.

Aged↗

Stimulation of neurotrophic factor secretion from 1321N1 human astrocytoma cells by novel diterpenoids, scabronines A and G.

Glial cells release neurotrophic factors that maintain neurons functionally. When rat pheochromocytoma cells (PC-12) were cultivated with the conditioned medium of human astrocytoma cells (1321N1) incubated with the new diterpenoids, scabronines A and G, isolated from Sarcodon scabrosus, they changed their morphology and there was neurite outgrowth. The scabronines increased the expression of mRNA for nerve growth factor (NGF), and the secretion of NGF from 1321N1 cells in a concentration-dependent manner. However, the enhanced neurite outgrowth produced by the conditioned media was slightly inhibited by NGF neutralizing antibody, and the concentration of NGF released in response to the scabronines was insufficient to cause differentiation. These results suggest that scabronines cause the secretion of other factors together with NGF from 1321N1 cells. The diterpenoids are useful drugs to clarify the mechanism of synthesis and secretion of neurotrophic factors.

Animals↗

Maitotoxin-induced nerve growth factor production accompanied by the activation of a voltage-insensitive Ca2+ channel in C6-BU-1 glioma cells.

1. The aim of the present study was to determine the effects of maitotoxin on nerve growth factor production and the Ca2+ influx in clonal rat glioma cells (C6-BU-1). 2. Maitotoxin (1 - 10 ng ml-1) induced a profound increase in 45Ca2+ influx in an extracellular Ca2+-dependent manner. However, high KCl had no effect at all. These effects were supported by the results from the analysis of intracellular Ca2+ concentration using fura 2. 3. The maitotoxin-induced 45Ca2+ influx was inhibited by inorganic Ca2+ antagonists, such as Mg2+, Mn2+ and Co2+. The inhibitory effect of Co2+ was antagonized by increasing the extracellular Ca2+ concentrations. 4. Maitotoxin (3 ng ml-1) as well as A-23187 (1microM) and dibutyryl cyclic AMP (0.5 mM) caused an acceleration of nerve growth factor (NGF) production in C6-BU-1 cells, as determined by NGF enzyme immunoassay. 5. Reverse transcription polymerase chain reaction (RT - PCR) analysis showed that maitotoxin (10 ng ml-1) enhanced the expression of NGF mRNA, which was abolished by the removal of extracellular Ca2+. A-23187 also accelerated its expression. 6. These results suggest that maitotoxin activates a voltage-insensitive Ca2+ channel and accelerates NGF production mediated through a Ca2+ signalling pathway in C6-BU-1 glioma cells.

Animals↗

[Changes in cytoskeletal proteins in childhood cataract lenses].

PURPOSE: Approximately 50% of congenital and childhood cataract seen in the clinic is of undetermined origin. Biochemical analysis of the cataracts is rare. This study analyzes lens proteins to determine the mechanism of congenital and childhood cataracts. METHOD: We analyzed the lens proteins from 10 young patients after cataract operations, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), densitometry analysis, and western immunoblotting. RESULTS: Densitometry of separated proteins showed a decrease in the high molecular protein bands of posterior subcapsular cataract (PSC). Specifically, spectrin (235 kDa), filensin (100 kDa), and vimentin (57 kDa) were absent from the SDS-PAGE of PSC. Increases in filensin and vimentin were observed in a Christmas tree cataract and lamellar cataracts. Western immunoblots confirmed the densitometry of SDS-PAGE. CONCLUSION: These results suggest that changes in cytoskeletal proteins may contribute to congenital and childhood cataract.

Adolescent↗

A new factor derived from 1321N1 human astrocytoma cells causes differentiation of PC-12 cells mediated through mitogen-activated protein kinase cascade.

Glial cells play an important role in maintaining neural function. In the present study, we examined the effects of a factor derived from human astrocytoma cells (1321N1) on differentiation of rat pheochromocytoma cells (PC-12). The conditioned medium which had been used for culture of 1321N1 cells caused the differentiation of PC-12 cells, suggesting that 1321N1 cells release a neurotrophic factor. The factor was apparently distinct from well-known neurotrophic factors, such as nerve growth factor (NGF), since it was resistant to boiling and trypsin treatment. The molecular size of the factor was assumed to be below 1000 through dialysis and ultrafiltration experiments. Furthermore, PC-12 cells were differentiated synergistically by the combined addition of NGF and the conditioned medium of 1321N1 cells. Partially purified fraction of the factor by Sephadex G-15 gel filtration column caused the prolonged activation of mitogen-activated protein kinase (MAPK). The differentiation of PC-12 cells induced by the fraction or NGF disappeared after the treatment with PD98059, a specific inhibitor of MAPK kinase (MEK), suggesting the involvement of MAPK in the differentiation. These results suggest that the new low-molecular factor derived from glial cells causes differentiation of PC-12 cells mediated through an activation of MAPK.

Animals↗

Heat influences on plasma insulin and glucagon in response to secretogogues in non-lactating dairy cows.

Four non-lactating cows were offered a maintenance diet of hay wafer and a commercial concentrate. They were housed in a thermoneutral (TN; 20 degrees C) and then a hot (30 degrees C) environment in an artificial climate chamber. Glucose, arginine, butyrate, and insulin were administered through one jugular catheter, and from a catheter on the other side venous blood was collected. The peak increments in plasma insulin after the glucose and butyrate administrations were lower during heat exposure. The response of insulin after arginine injection was smaller in the hot compared with the thermoneutral environment; however, arginine injection resulted in a significantly higher secretion of glucagon in the hot environment. The response area of insulin after the insulin injection was smaller in the hot environment; however, insulin clearance rate was not changed. It is concluded that in non-lactating cows, insulin release is probably lower during heat exposure. With respect to plasma glucose during heat exposure, the lower basal values, lower concentrations after the end of the glucose infusion, and delayed recovery to basal values after the butyrate and insulin administrations observed, may indicate lower gluconeogenesis and glycogenolysis in the hot environment.

Animals↗

Effects of heat exposure on plasma insulin, glucagon and metabolites in response to nutrient injection in heifers.

Effects of heat exposure on plasma insulin, glucagon, and metabolite responses following injection of various nutrients were investigated in heifers. Four heifers, fed hay wafer and a commercial concentrate, were exposed to thermoneutral (20 degrees C) and hot (30 degrees C) environments. Glucose, arginine and butyrate (each injection at 0.625 mmol/kg) and insulin (0.2 U/kg) were injected intravenously, and then blood samples were collected at regular intervals through jugular vein catheters. Insulin secretion in response to glucose and arginine injection was not affected by heat exposure. However, the insulin response following butyrate injection was inhibited in heifers exposed to heat. In the hot environment, glucagon responses following the arginine and butyrate injections were augmented significantly, however glucagon levels were inhibited following the glucose injection. It is concluded that heat stress causes an inhibition of the insulin response to butyrate injection, and an increase in the glucagon response following arginine and butyrate injection. Plasma metabolite concentrations altered in accordance with the changes in the concentration of pancreatic hormones.

3-Hydroxybutyric Acid↗

Maternal Nanos regulates zygotic gene expression in germline progenitors of Drosophila melanogaster.

Maternal Nanos (Nos) protein is required for germline development in Drosophila embryos. Here we show that Nos regulates zygotic gene expression in the germline progenitors, or pole cells. In order to probe the gene expression in pole cells, we screened ten enhancer-trap lines which showed beta-gal expression in pole cells. All of these enhancer-trap markers were fully activated in pole cells after their migration to the embryonic gonads. In the pole cells lacking Nos, the expression of nine out of ten enhancer-trap markers was affected. Among nine markers, five (Type-A) were prematurely expressed in the pole cells during the course of their migration. The expression of other four markers (Type-B) initiated correctly after pole-cell migration, but their expression was significantly reduced. Thus, we conclude that the maternal Nos plays a dual role in zygotic gene regulation in pole cells: to define the stages of expression for Type-A markers, and to enhance expression for Type-B markers. Contrary to our results, "Heller and Steinmann-Zwicky (1998)" have recently reported that no premature expression of Type-A markers occurs in the pole cells of embryos derived from nos mutant females. This discrepancy is due to the difference in the nos mutant alleles used for these analyses. We used the much stronger allele, nosBN.

Animals↗

Insulin-independent glucose uptake in growth hormone treated dairy cows.

Our aim was to determine the effect of growth hormone on non-insulin-mediated glucose disposal in lactating dairy cows. Following 5 d of subcutaneous injections of either saline or growth hormone, insulin, somatostatin or insulin plus somatostatin were infused for 2 h each, in a series of experiments. Coincident with this, unlabelled glucose was infused at a variable rate to maintain a constant plasma glucose concentration. Glucose, doubly labelled with deuterium, was also infused for the calculations of glucose turnover. Plasma insulin levels were reduced to nearly zero by the infusion of somatostatin; under such conditions whole body glucose disposal should be non-insulin-mediated. Dairy cows treated with growth hormone, which had significantly increased milk yields on the day before the experimental infusions, did not have different levels of whole body non-insulin-mediated glucose disposal when expressed in absolute terms. Growth hormone did not affect non-mammary non-insulin-mediated glucose uptake estimated by calculation. Growth hormone significantly inhibited insulin-mediated glucose uptake when plasma insulin levels were elevated. Glucose uptake during insulin plus somatostatin infusion was not significantly different from that of the insulin only infusion.

Animals↗

Insulin and glucagon secretion in lactating cows during heat exposure.

Heat stress affects endocrine systems in cows. This study investigated changes in insulin and glucagon secretion between thermoneutral (TN; 18 degrees C, relative humidity [RH] 60%) and hot (28 degrees C, RH 60%) environments in lactating cows. Glucose, arginine, and butyrate were administered i.v. to four cows (mean, at 83 d postpartum) in each environment. Blood was collected via a jugular catheter at regular intervals. Heat exposure resulted in a marked increase in respiration rate and rectal temperature. A decrease in milk yield was also observed during heat exposure. Basal insulin concentrations were elevated, and basal glucose concentrations tended to be lower in the hot environment. Peak values of insulin and glucagon following the arginine injection were significantly higher in the hot than in the TN environment. The insulin peak value in response to the butyrate infusion was also higher during the heat exposure. However, insulin and glucagon responses to the glucose load were not affected by heat stress. The increase in plasma glucose concentration following arginine injection was inhibited by the heat exposure. In conclusion, heat stress resulted in a higher insulin secretion in lactating cows. Glucagon secretion in response to the arginine injection was enhanced, but the rise in plasma glucose was inhibited by heat exposure. These changes would be related to a reduction in milk yield during heat stress.

Animals↗

[A case of the double mitral valve orifices].

In 1996 student course of gross anatomy dissection at Iwate Medical University School of Medicine, a case of the double mitral valve orifices was found in a heart of a 73-year-old male cadaver who died of acute pneumonia. No other congenital anomalies were detected in this heart. There was no prior history of heart disease. The left atrioventricular ostium was an oval with a long axis of 4 cm and a short axis of 2.5 cm. There were double mitral valve orifices, main and accessory (Fig. 1). The main orifice situated left ventrally with a long axis of 2.5 cm and a short axis of 1.7 cm, had a larger cleft. The accessory orifice situated right dorsally with a long axis of 2.5 cm and a short axis of 1.2 cm, had a smaller cleft. There was a bridge-like valvular leaflet between the two clefts (Fig. 1). Disregarding small incisions, both the main and the accessory mitral valves had two cusps, anterior and posterior. The anterior and posterior cusps of the main and accessory valves were continuous and considered to be homologous with the anterior and posterior cusps of the normal mitral valve. In other wards, this double mitral valve orifices was appeared to be formed by bridging the cleft between the anterior and posterior cusps of the normal mitral valve. The valvular cusps of the main or the accessory mitral valve had proper chordae tendineae and papillary muscles. The papillary muscles were classified into four groups, anterior (A), anterolateral (AL), posterior (P) and posterolateral (PL) according to Taniya (1974). The anterior papillary muscles group had a base at the anterior wall of the left ventricle, the anterolateral papillary muscles group had at the whole lateral wall, the posterior papillary muscles group had at the boundary between the interventricular septum and the posterior wall, and the posterolateral papillary muscles group had at the posterior part of the lateral wall (Fig. 3). The anterior and anterolateral papillary muscles groups, and the posterior and posterolateral papillary muscles groups of Taniya (1974) constituted the anterior and the posterior papillary muscles respectively in the general use of the anatomical term. The chordae tendineae attached to the cusps of the main orifice arose from the anterior, anterolateral and posterior papillary muscles groups (Fig. 2a). And those attached to the cusps of the accessory orifice arose from the posterolateral papillary muscles group (Fig. 2b). The chordae tendineae were also classified into three types after Taniya (1974). The A type chordae tendineae were thick and attached to the base of the cusps, the type B were moderately thick and attached to the midway between the base and the margin of the cusps, and the C type were thin and attached to the margin of the cusps. The main mitral valve had 6 type A, 8 type B and 40 type C chordae. The accessory mitral valve had 0, 6 and 30 chordae respectively (Fig. 2). This case of the double mitral valve orifices is considered to be included in the central type of Cascos et al. (1967) and the bridge type of Elfenbein et al. (1967). A morphogenetic mechanism of this type of the double mitral valve orifices is explained by an abnormal adhesion between the dorsal endocardial cushion and the left lateral endocardial cushion like mass in early developmental stage according to Wimsatt and Lewis (1948). But a possibility of an abnormal orifice in the posterior cusp which is produced in the process of formation of cusps, chordae tendineae and papillary muscles in later stage, still remains to be discussed.

Aged↗

[Relationship between vascular endothelial growth factors and advanced glycation end products in the human vitreous].

The relation between vascular endothelial growth factor (VEGF) and advanced glycation end product (AGE) is considered a primary factor in the development of diabetic retinopathy. Regarding the relation between VEGF in the vitreous body and pentosidine, an AGE, we compared a diabetic (DM) group (7 eyes) with a nondiabetic (nonDM) group (7 eyes), and investigated the correlation between VEGF and pentosidine by calculating the correlation coefficient. Levels of both VEGF and pentosidine were significantly higher in the DM group (p < 0.01, p < 0.05), and a positive correlation was observed between the levels of VEGF and pentosidine (r = 0.770, p < 0.001). Since it is clear that there is a relation between VEGF and pentosidine in the vitreous body, we speculated that AGE is related to the secretion of cytokine in patients with diabetic retinopathy, and that it affects the development and progression of the disease.

Adult↗