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Biomedical subjects

N Yoshimura

Publications and source records attributed to N Yoshimura.

At least 415 records · Page 23Linked to original sources

Identification of transforming growth factor-beta expressed in cultured human retinal pigment epithelial cells.

PURPOSE: This study examined whether retinal pigment epithelial (RPE) cells have the capacity to express transforming growth factor-beta (TGF-beta). Also examined were TGF-beta isoform genes expressed in the RPE cells. METHODS: Complementary DNA (cDNA) was generated from polyA+ RNA extracted from human RPE cells in culture, and polymerase chain reaction (PCR) using degenerate and specific primers of the known TGF-beta isoforms was carried out by using the cDNAs as templates. Sequencing and Southern blot analysis of the PCR products, and Northern blot analysis were performed to identify which isoforms are expressed in RPE cells. RESULTS: PCR using degenerate primers showed that most of the amplified sequences were derived from TGF-beta 2, although TGF-beta 1 expression was shown by using specific primers. Northern blot analyses confirmed not only the expression of the TGF-beta gene, but also suggested that alternative splicing of TGF-beta mRNA in the RPE cells occurred. CONCLUSION: Cultured human RPE cells expressed TGF-beta 1 and beta 2 genes, whereas gene expression of TGF-beta 3 was not confirmed. Our data suggest that RPE cell are intraocular origin of TGF-beta production.

Amino Acid Sequence↗

Synergistic suppression of retinal pigment epithelial cell proliferation in culture by radiation and hyperthermia.

PURPOSE: To study combination effects of radiation and hyperthermia on the in vitro cell proliferation of cultured human retinal pigment epithelial cells; to explore possible application of the combination treatment for proliferative vitreoretinopathy. METHOD: Cultured human retinal pigment epithelial cells were treated by radiation, hyperthermia, or a combination of the two. Cell proliferation was evaluated by counting the cell number and by the uptake of bromodeoxyuridine. RESULTS: X-ray irradiation of 100 cGy or 300 cGy was not effective in suppressing proliferation of the retinal pigment epithelial cells. Similarly, heat treatment at 42 degrees C or 43 degrees C for 30 min did not suppress proliferation of the retinal pigment epithelial cells. However, combination of hyperthermia at 42 degrees C for 30 min with 300 cGy irradiation suppressed cellular growth of the retinal pigment epithelial cells to 35.7% of the control as estimated by the cell counting and to 48.3% by bromodeoxyuridine uptake study. Combination treatment of 43 degrees C, 30 min hyperthermia and 300 cGy irradiation was more effective. CONCLUSIONS: A combination of low-dose radiation and mild hyperthermia is effective in the suppression of growth of cultured human retinal pigment epithelial cells, and the effects were found to be synergistic. It is expected that the synergistic effects will lower the radiation dose and also reduce the possible side effects of radiation in the treatment of proliferative vitreoretinopathy.

Cell Count↗

Platelet-derived growth factor gene expression in cultured human retinal pigment epithelial cells.

Gene expression of platelet-derived growth factor (PDGF) and its receptors in cultured human retinal pigment epithelial (RPE) cells was studied by using semiquantitative polymerase chain reaction. The RPE cells were found to express PDGF A- and B-chain genes as well as alpha- and beta-receptor genes with dominant expression of B-chain and beta-receptor isoforms. Phorbol myristate acetate (PMA) and thrombin increased the expression of PDGF B-chain gene to 19.8 +/- 1.75 and 15.9 +/- 1.84 fold (n = 3) of the control without affecting beta-receptor gene expression. PDGF produced by the RPE cells may play an important role in the pathogenesis of some ocular proliferative diseases.

Base Sequence↗

Tumor necrosis factor-alpha gene is expressed in stimulated retinal pigment epithelial cells in culture.

Expression of TNF-alpha gene in the retinal pigment epithelial (RPE) cells was studied by using polymerase chain reaction (PCR). PCR for human TNF-alpha gene using complementary DNA (cDNA) of control RNA prepared from non-stimulated RPE cells failed to show expression of TNF-alpha gene. However, PCR by using cDNA prepared from interleukin-1 beta (IL-1 beta)-stimulated RPE cells revealed expression of the gene, suggesting in vivo production of TNF-alpha from RPE cells in response to IL-1 beta.

Base Sequence↗

Cardiac metastasis of lung cancer. A study of metastatic pathways and clinical manifestations.

BACKGROUND: Although lung cancer frequently spreads to the heart, details of cardiac metastases of lung cancer have not been fully discussed. The authors attempted to elucidate the relationship between the mechanisms of cardiac metastasis and a variety of clinical manifestations caused by cardiac metastasis. METHODS: Clinical and autopsy records were reviewed in 74 autopsied cases of lung cancer. In cases with cardiac metastasis, the metastatic pathways to the heart were determined by the macroscopic examinations, and the relationship between the metastatic pathways and the clinical manifestations were studied. RESULTS: Metastases to the pericardium or heart were seen in 23 cases (31%). A lymphatic metastatic pathway was detected in 18 cases (hilar lymphatic routing in 12 cases, and mediastinal lymphatic routing in 6 cases), and a hematogenous metastatic pathway was detected in 5 cases. Malignant pericardial effusion was documented in 15 of 23 cases. The metastatic pathway in 14 of 15 cases was lymphatic (hilar lymphatic routing in 10 cases, and mediastinal lymphatic routing in 4 cases). Patients showing lymphatic metastasis had higher incidence of malignant pericardial effusion than those with hematogenous metastasis (P less than 0.05). Of 23 cases of cardiac metastasis, myocardial infarction was found in 1 case, resulting from the compression of the coronary arteries by the tumor. Concurrent supraventricular arrhythmias were recorded in eight cases with cardiac metastasis. Patients with cardiac metastasis had higher incidence of arrhythmia than those without cardiac metastasis (P less than 0.05). In cases of cardiac metastasis, patients with arrhythmia were older (P less than 0.01) than those without arrhythmia. CONCLUSIONS: The authors concluded that the hilar lymphatic pathway is essential for early development of malignant pericardial effusion in lung cancer and that aging and cardiac metastasis may be responsible for arrhythmia in patients with lung cancer.

Age Factors↗

Neuropeptide-induced cytosolic Ca2+ transients and phosphatidylinositol turnover in cultured human retinal pigment epithelial cells.

Neuropeptide-induced mobilization of cytosolic free Ca2+ concentration ([Ca2+]i) and phosphatidylinositol (PI) turnover in cultured human retinal pigment epithelial (RPE) cells were studied and their temporal relationship was compared. After RPE cells were loaded with fura-2/AM, [Ca2+]i was analyzed using a digital imaging microscopy system. Bombesin-related peptides which include bombesin, neuromedin B, and neuromedin C induced significant [Ca2+]i transients in RPE cells, whereas other neuropeptides, neuropeptide Y, vasoactive intestinal polypeptide (VIP), and substance P were not effective to produce [Ca2+]i transients. The percentage of reactive cells which showed positive [Ca2+]i transients induced by bombesin-related peptides was around 50%. Bombesin (1 microM) showed a peak concentration of 663 +/- 27.0 nM (mean +/- S.E.M., n = 61), neuromedin B (1 microM), 327 +/- 28.7 nM (mean +/- S.E.M., n = 38), and neuromedin C (1 microM), 357 +/- 22.7 nM (mean +/- S.E.M., n = 32). Ca2+ transients occurred within 30 s and lasted less than 5 min after the application of the neuropeptides. Chelation of the extracellular Ca2+ by EGTA significantly shortened the total time of [Ca2+]i transients induced by the above. The measurements of phosphoinositides in RPE cells revealed that neuropeptide-induced PI turnover was as quick as [Ca2+]i transients. Inositol biphosphate (IP2) and inositol triphosphate (IP3) in RPE cells showed transient increases at 15 s after the stimulation by bombesin-related peptides. These data show that changes in [Ca2+]i and PI turnover are directly linked and both are important in the signal transduction system of bombesin-related peptides in RPE cells. The data also suggest that bombesin-related peptides may play some possible roles in RPE cells.

Calcium↗

Clinical factors which influence the long-term survival of kidney allografts donated from haploidentical donors.

All patients with renal transplants are very much concerned about their chance of long-term graft function. Chronic rejection is the most common cause of decline in function and graft failure, and after the first post-transplant year, 3-4% of recipients lose their graft every year. However, the cause, time of onset and mechanism of decline in graft function are not clear. Also unclear is whether clinical and laboratory parameters may predict patients who are at risk of developing chronic rejection. The aim of this study was to find the marker which may determine long-term graft survival in the azathioprine era and in the cyclosporine era.

Biomarkers↗

Pathological features of the pulmonary artery in Takayasu arteritis.

Little attention has been paid to the pathological features of the pulmonary artery in Takayasu arteritis. Autopsy specimens of 6 cases of this disease were studied. Lesions were found in the aortic arch and its brachiocephalic branches in all cases and in both the aortic arch and thoracoabdominal aorta in 5 cases. The pathohistologic characters of the pulmonary artery were very similar to those of the systemic artery. Stenosis-recanalization, so-called blood vessels-in-blood vessels, of the pulmonary elastic arteries were found in four cases. These lesions were not observed in the systemic arteries, and most of the newly formed channels in them seemed to be branches of bronchial arteries. Luminal obstruction of pulmonary muscular arteries was observed in 4 cases, cellular arteritis of muscular arteries in 2 cases, and angiomatoid dilatation of small blood vessels in 2 cases. Thus in this study we found peculiar stenosis-recanalization lesions of the pulmonary elastic arteries, and also showed that the pulmonary elastic and muscular arteries are frequently involved in Takayasu arteritis. These findings suggest that pulmonary hypertension could influence morbidity and long-term mortality in this disease.

Adult↗

Changes in erythrocyte membrane fluidity by endotoxin in rats.

The effect of endotoxin on fluidity and lipid composition of the erythrocyte membrane was studied in rats following the intraperitoneal administration of endotoxin (30 mg.kg(-1) body weight). Erythrocyte membrane fluidity measured with 16-stearic acid spin label (16-SAL) was significantly decreased in the endotoxin-treated rats as compared with control. A decrease of lysophosphatidylcholine in the membrane lipid was evident in the endotoxin-treated rats. The cholesterol to phospholipid molar ratios and other phospholipid fractions did not differ significantly in the two groups. The levels of plasma Beta-glucuronidase activity and lipoperoxide were significantly increased in the endotoxin-treated rats when compared to controls. There were significant correlations between the parameter of 16-SAL in erythrocytes and plasma Beta-glucuronidase activities or lipoperoxide from both endotoxin-treated and control groups, P < 0.005 or P < 0.02 respectively. In conclusion endotoxin decreased rat erythrocyte membrane fluidity in vivo. Since membrane fluidity is closely related to the vital functions of the membranes, the change described could be related to the abnormality of cell membrane functions in endotoxin shock state.

Journal Article↗

Systemic atrioventricular valve replacement in an infant with corrected transposition of the great arteries.

A 3-month-old infant with corrected transposition of the great arteries and severe systemic atrioventricular valve regurgitation due to "Ebstein-like anomaly" is reported. Through a right thoracotomy and longitudinal left atrial incision, a 19-mm St. Jude Medical valve was implanted into the annulus without removing the native valve. He is doing well 7 months after operation.

Ebstein Anomaly↗

The effects of pravastatin on hyperlipidemia in renal transplant recipients.

Hyperlipidemia may be one of the risk factors in the development of atherosclerotic disease in renal transplant recipients. In the present study, 24 kidney recipients with hyperlipidemia were treated with an HMG-CoA reductase inhibitor, pravastatin (10 mg/day). All recipients had been treated with cyclosporine (CsA), azathioprine (Az), and prednisolone (Pred). The mean total cholesterol (T-chol) level decreased from 323 +/- 7.4 to 261 +/- 7.9 mg/dl at one month after starting treatment (P less than 0.01) and this level did not change during treatment for further 6 months. The mean LDL cholesterol level was also decreased from 205.9 +/- 11.2 to 118.7 +/- 8.1 mg/dl at 3 months after starting treatment (P less than 0.01). On the other hand, pravastatin did not affect the levels of HDL-cholesterol and triglycerides. Pravastatin did not show any effects on the white blood cell, monocyte, and lymphocyte counts, or the hemoglobin concentration (NS). One patient displayed a slight elevation of aspartate aminotransferase and alanine aminotransferase levels, but this was not sufficient to cease treatment. Pravastatin did not adversely affect the renal function or creatinine phosphokinase (CPK) levels. Two recipients developed nausea and vomiting and their treatment was stopped. Pravastatin appears to be a safe and efficacious method of treating hyperlipidemia in renal transplant recipients.

Cholesterol↗

Diffuse Mallory bodies in the liver, diffuse Lewy bodies in the brain and diffuse fat replacement (lipomatous pseudohypertrophy) of the pancreas in a patient with juvenile Parkinson's disease.

A 38-year-old male patient with the juvenile variant of Parkinson's disease, in whom onset had occurred at the age of 24 yr, was autopsied. There were no clear symptoms of pancreatic or hepatic insufficiency during the entire clinical course. The only notable features were a slightly delayed decrease of the blood glucose level in an oral glucose tolerance test, slightly elevated levels of serum alkaline phosphatase and serum lactate dehydrogenase, and episodic loose stools. Autopsy revealed uniform enlargement of the pancreas due to massive fat replacement (lipomatous pseudohypertrophy): the exocrine glandular elements showed marked atrophy and loss, while the islets of Langerhans were preserved. The liver exhibited a histology closely mimicking alcoholic hepatitis associated with the diffuse presence of Mallory bodies (MBs), possibly indicative of a disturbance of protein metabolism. The nervous system showed the diffuse presence of Lewy bodies (LBs) in the cerebrum in addition to the ordinary lesions of Parkinson's disease. Although the etiopathogenesis of none of these three lesions has been well elucidated, common epitopes of MBs and LBs have recently been demonstrated. Therefore, the present case study suggests that a specific underlying toxic agent may cause diffuse LBs in the brain on the one hand, and diffuse MBs in the liver and lipomatous pseudohypertrophy of the pancreas on the other.

Adult↗

Wegener's granulomatosis. Associated with diffuse pulmonary hemorrhage.

The authors report a case of Wegener's granulomatosis with the unusual manifestation of diffuse pulmonary hemorrhage. A 58-year-old man complained of bloody sputum and fever. Chest X-ray films showed multiple nodular shadows in both lung fields. He was diagnosed as having Wegener's granulomatosis by transbronchial lung biopsy, which revealed necrotizing granulomatous inflammation with necrotizing vasculitis. Despite treatment with cyclophosphamide and prednisolone, his condition rapidly deteriorated. An extensive diffuse alveolar shadow appeared in both lung fields in chest X-ray films, anemia became worse, and he died of respiratory failure. Autopsy revealed diffuse alveolar hemorrhage with necrotizing capillaritis in addition to the typical pathological findings in Wegener's granulomatosis. The capillaritis was characterized by neutrophilic infiltration of alveolar septa, and fibrin thrombi in alveolar capillaries. Diffuse pulmonary hemorrhage is uncommon in Wegener's granulomatosis. However, once diffuse pulmonary hemorrhage occurs, the respiratory condition rapidly deteriorates and is life-threatening. Therefore, accurate diagnosis and appropriate treatment are required.

Granulomatosis with Polyangiitis↗

Immunohistochemical localization of c-myc oncogene product in oral papilloma.

The expression of c-myc oncogene products in oral papillomas was studied by using an immunohistochemical method. The oncogene products were detected in 17(70.8%) of the 24 oral papillomas under study. The expression of the products was evaluated, and the histologic localization in proliferating epithelial cells of the oral papillomas was determined. In the basal cell layer, the products were detected in the nuclei of 16 oral papillomas, and in the cytoplasm of 12 oral papillomas. In the nuclei of cells in the spinous cell layer, the products were detected in 4 oral papillomas, and in the cytoplasm of 13 oral papillomas. In the keratinized cells, the products were not detected in the nuclei, but they were identified in the cytoplasm of three oral papillomas. The results suggested that the c-myc oncogene product might play an important role for proliferation and differentiation of the oral papilloma.

Adolescent↗

Distribution of two types of infiltrating helper T cells among rat renal allografts, spleen, peripheral blood, and regional lymph node cells.

Cytokine production by graft-infiltrating cells (GIC) of helper T cell (Th) subtypes were investigated in rat kidney allograft compared to those infiltrating the spleen, those in peripheral blood, and in regional lymph nodes. The relative proportion of OX-22 positive cells (Th type 1) and the W3/25-positive cells (Th) of GIC was 49.4 +/- 2.1% on day 3 and 52.7 +/- 2.2% on day 6, whereas those in spleen cells was 80.3 +/- 4.1% (p less than 0.05) and 95.5 +/- 2.5% (p less than 0.01), respectively. GIC produced more B cell-stimulating factor 2 (BSF-2, 106.3 +/- 8.2 U/ml) when compared to spleen cells (15.9 +/- 4.6 U/ml; p less than 0.01), peripheral blood mononuclear cells (4.2 +/- 1.0 U/ml; p less than 0.01) or lymph node cells (24.9 +/- 4.0 U/ml; p less than 0.01) upon stimulation with donor BN lymphocytes in vitro. However, GIC produced less interleukin 2 (IL-2, less than 1.0 U/ml) and IL-3 (less than 1.0 U/ml) than produced by spleen cells (5.1 +/- 0.9 U/ml, p less than 0.05, and 10.5 +/- 2.3 U/ml, p less than 0.01, respectively) or lymph node cells (4.5 +/- 0.4 U/ml, p less than 0.01; 23.5 +/- 3.1 U/ml, p less than 0.01, respectively). These results demonstrate that the subtype of Th cells which produce BSF-2 migrates selectively to allografts.

Animals↗

Mass cultivation of human retinal pigment epithelial cells with microcarrier.

Microcarrier cell culture permits mass cultivation of anchorage-dependent cells. In this study, mass cultivation of human retinal pigment epithelial (RPE) cells was studied using Cytodex 3 (Pharmacia) as a microcarrier. Human RPE cells were established from aborted fetuses and cultured in Dulbecco's modified Eagle's Medium (DMEM). After the 3rd-5th passages, RPE cells were suspended in 50 ml of DMEM in a spinner flask at a density of 2 x 10(5)/ml, and Cytodex 3 was added to the spinner flask at a bead density of 10 mg/ml. Cultures were maintained at 20-50 rpm (final speed) on a magnetic stirrer, and DMEM was added up to 100 ml. Fifty milliliters of DMEM were decanted and replaced with fresh DMEM every 2 days. After 1 week, a cell density of 10(6)/ml DMEM was obtained. Phase contrast microscopy showed bridging formation between microcarriers, which suggests tight cell adhesion. Microcarrier cell culture has a variety of advantages which include greater cell production, use of less medium and less risk of contamination compared to the conventional monolayer culture technique, and it also allows passaging without using proteases. Using this culture system, greater possibilities for wider application of new cell cultures can be expected.

Cell Count↗

Acute relapsing sensory neuropathy associated with IgM antibody against B-series gangliosides containing a GalNAc beta 1-4(Gal3-2 alpha NeuAc8-2 alpha NeuAc)beta 1 configuration.

We report a patient with a relapsing form of the acute sensory neuropathy syndrome associated with IgM-kappa type monoclonal gammopathy of undetermined significance. He rapidly developed marked sensory ataxia without weakness following an upper respiratory tract infection at age 44. The symptoms reached their maximum in a few days, followed by subsequent gradual improvement over a few weeks. However, unsteady gait remained as a chronic deficit. Stepwise progression of his symptoms occurred over 15 years with 10 similar relapses. Sensory nerve conduction studies showed the absence of action potentials, and sural nerve biopsy revealed the marked loss of large myelinated fibers. The patient's serum had an extremely high titer of an IgM monoclonal antibody directed against gangliosides GD2, GD1b, GT1b, and GQ1b.

Acute Disease↗

[Inhibitory effects of Hachimijiogan on micturition reflex via the locus coeruleus].

Pharmacological studies were undertaken to elucidate the role of Hachimijiogan in the micturition reflex via the locus coeruleus, using alpha-chloralose-anesthetized cats. Rhythmic contractions of the urinary bladder induced by continuous infusion of saline into the bladder were dose-dependently inhibited by intravenous injection of Hachimijiogan (10, 30 and 90 mg/kg), as well as flavoxate hydrochloride (1 and 3 mg/kg). In contrast, contraction of the urinary bladder elicited by electrical stimulation of the locus coeruleus was significantly suppressed by intravenous injection of flavoxate, but not affected by that of Hachimijiogan. These results suggest that Hachimijiogan acts on the afferent pathway from the urinary bladder to the locus coeruleus, thereby inhibiting the micturition reflex, while it has no effects on the efferent pathway from the locus coeruleus to the urinary bladder.

Animals↗