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Y Uchiyama

Publications and source records attributed to Y Uchiyama.

At least 19 recordsLinked to original sources
PubMed1992

Structural organization of the gene encoding rat cystatin beta.

A genomic DNA clone encompassing the gene (cy beta) encoding rat cystatin beta (Cy beta) was isolated by screening with a rat cy beta cDNA as a probe. The gene spans about 2.6 kb and comprises three exons. The first intron is located between Lys22 and Val23 and the second between Lys56 and Val57 in the deduced amino acid sequence of Cy beta. The second exon contains the highly conserved QVVAG sequence which, unlike the sequence of other cystatin family members, is not split by an intron. In the 5'-upstream region, three SP-1-binding sites exist, but no typical TATA-box or CAAT-box sequences are found. The difference in the organization of the rat cy beta gene, encoding a family-1 cystatin, from that encoding members of the other cystatin families, suggests that cy beta diverged from a common ancestral gene earlier than the separation of genes encoding family-2 and family-3 cystatins.

Amino Acid Sequence↗
PubMed1992

Variations in immunoreactivity for phenobarbital- and 3-methylcholanthrene-inducible cytochromes P-450, and NADPH-cytochrome P-450 reductase in rat liver over twenty-four hours.

To reveal distribution patterns of phenobarbital- and 3-methylcholanthrene-inducible cytochromes P-450 (PB and MC) and NADPH-cytochrome P-450 reductase (P-450red) within the liver acinus of untreated rats, and their variations over 24 h, hepatic samples were examined by immunohistochemistry and image-analyzer at evenly spaced six time points over 24 h. When examined in semi-thin sections obtained from Epon-embedded, freeze-dried, and paraformaldehyde vapor-phase fixed materials, the immunoreactivity for these enzymes showed different distribution patterns within the liver acinus. Immunodeposits for PB were predominantly distributed in perivenous hepatocytes, whereas those for MC and P-450red were slightly more intense in periportal hepatocytes at each time point. The immunoreactivity for PB and MC in both perivenous and periportal hepatocytes increased during the dark period, peaking early in the light period. These variations coincide well with our previous morphometric results (Uchiyama and Asari, 1984); the volume and surface densities of rough endoplasmic reticulum (rER) in hepatocytes increased during the dark period. On the other hand, weak fluctuation was demonstrated in the immunoreactivity for P-450red in hepatocytes of both zones. These results suggest that PB and MC are retained in rER rather than smooth endoplasmic reticulum (sER) of hepatocytes obtained from untreated rats. These enzymes in sER may be short in their half-life spans.

Animals↗
PubMed1992

The effect of Ninjinyoeito on Werner's syndrome skin fibroblasts.

The effect of Ninjinyoeito on three cases of Werner's syndrome fibroblasts was investigated. In all three cases, groups which were treated with Ninjinyoeito showed improved life-span doubling levels compared to groups which were not treated with Ninjinyoeito. Also, all of the treated group in all three cases showed significantly higher values in the rates of DNA synthesis including two cases which showed significantly higher rates of protein synthesis.

Adult↗
PubMed1992

Intra- and extracellular localization of hyaluronic acid and proteoglycan constituents (chondroitin sulfate, keratan sulfate, and protein core) in articular cartilage of rabbit tibia.

To demonstrate the intra- and extracellular localization of hyaluronic acid (HA) in articular cartilage of the rabbit tibia, biotinylated HA binding region, which specifically binds to the HA molecule, was applied to the tissue. In comparison with the localization of HA, that of chondroitin sulfate (CS), keratan sulfate (KS), and the protein core (PC) of the proteoglycan was examined by immunohistochemistry. Strong positive staining for HA was detected in chondrocytes located in the transition between the superficial and middle zones of the tissue. Pre-treatment with chondroitinase ABC, keratanase II, or trypsin enhanced the stainability for HA in peri- and intercellular matrices. Immunohistochemistry with or without enzymatic pre-treatment demonstrated that immunoreactivity for CS, KS, and PC was distinctly discerned in chondrocytes and in the extracellular matrix located in the middle and deep zones. In particular, the immunoreactivity for KS and PC was augmented by pre-treatment with chondroitinase ABC not only in chondrocytes but in the extracellular matrix located in the middle and deep zones. Microbiochemical analysis corresponded well with histochemical and immunohistochemical results. These results suggest that HA is abundantly synthesized and secreted in chondrocytes located in the transition between the superficial and middle zones.

Aggrecans↗
PubMed1992

Changes in subcellular structures of parietal cells in the rat gastric gland after omeprazole.

Omeprazole, an inhibitor of gastric acid secretion, was administered to rats at a dosage of 20 mg/kg/day for 14 and 35 days, and subsequent changes in subcellular structures of parietal cells were analyzed using morphometry and immunocytochemistry. Plasma gastrin levels were also examined, showing two times higher levels in the experimental groups than in the non-treated control. The volume and surface densities significantly decreased in tubulovesicles of the cells in the experimental rats. In the long term treatment of omeprazole (35 days), the volume density of microvilli on the membranes of secretory canaliculi in the cells also decreased significantly, whereas that of lysosomes clearly increased. By electron microscopy, many dense bodies of various shapes often appeared in the cytoplasm of parietal cells after the omeprazole treatment. Immunocytochemistry revealed that large granular immunodeposits for cathepsin B increase in the epithelial cells of the gastric glands after omeprazole treatment. These results suggest that omeprazole induces quantitatively significant decreases in both tubulovesicles and canalicular microvilli. The decreases in these membrane structures may possibly be ascribed to the degradation of the membrane in lysosomes; the proton pump on the membranes bound irreversibly with omeprazole is believed destined to be degraded in lysosomes.

Animals↗
PubMed1992

Localization of hyaluronic acid, chondroitin sulfate, and CD44 in rabbit cornea.

To demonstrate the localization of hyaluronic acid (HA) in rabbit cornea, the biotinylated HA-binding region, which specifically binds to the HA molecule, was applied to the tissue. Localization of chondroitin sulfate (CS) and CD44, a possible cell surface receptor for HA, were also examined by immunohistochemistry. The stainability of HA changed depending on the fixatives used. Reaction products for HA were distinctly detected in epithelial cells and stromal keratocytes, but faintly in the extracellular matrix of the stroma when unfixed cryosections were applied. No positive reaction was found in the endothelium, except that the positive deposits formed a continuous layer on the apical surface of the endothelium. Electron microscopy using samples fixed with 2% paraformaldehyde revealed gold particles indicating HA labeling the intercellular space of the epithelium and stromal extracellular matrix. No intracellular deposition was detected in epithelial cells, whereas the gold labeling was seen in vacuolar structures of stromal keratocytes. Immunodeposits for CS were intensely localized in the epithelium and stroma, and weakly in the endothelium. Immunoreactivity for CD44 was found in the epithelial, endothelial and stromal cells. In particular, immuno-deposits for CD44 were detected in basal parts of epithelial cells, while they were localized in the apical surface of endothelial cells. These results suggest that HA is synthesized in and secreted from epithelial and stromal cells of rabbit cornea, while the localization of HA in the apical surface of the endothelium is closely associated with that of CD44. Moreover, the presence of CS in corneal tissue may play a role in its transparency, as has been suggested for keratan sulfate and dermatan sulfate.

Animals↗
PubMed1992

Heterogeneity of monoclonal immunoglobulins with antistreptolysin-O activity detected in the cases of essential monoclonal gammopathy and multiple myeloma.

Two different types of monoclonal human immunoglobulins (M-components) with antistreptolysin-O (ASO) activity were investigated. The M-component FM with essential monoclonal gammopathy revealed to have an ASO activity, demonstrated not only by streptolysin-O neutralizing assay according to Ranz-Randall's method, but also by passive agglutination assays and precipitation on agar. The ASO activity was shown to reside in the Feb. These findings suggest that the M-component FM have a true antibody activity. On the other hand, ASO activity of M-component TT with multiple myeloma was detected only by streptolysin-O neutralizing assay, but the passive agglutinating assays and precipitation on agar showed no positive results. It has not been fully confirmed if the M-component TT behaves as a true antibody activity. Heterogeneity of the M-components with ASO activity was discussed.

Aged↗
PubMed1992

[A case of primary acinic cell tumor of the trachea].

A 62-year-old woman complained cough and hemosputum. Chest X-ray film and thoracic computed tomography revealed tumor shadow, localizing in the upper trachea, measuring 2 x 1 cm. Through a median sternotomy, resection of the trachea of 4 cartilagous rings was made, followed by end-to-end anastomosis. Fixing of tracheal anastomosis was performed for 2 weeks using SOMI brace. The size of tumor was 1.3 cm in diameter. Histologically, nodular tumor foci were seen close to the normal trachea gland, localing in tracheal mucosa. Tumor cells with basophilic cytoplasma developed acinical structure. Postoperative course was uneventful. This patient is alive 42 months after resection. This disease is extremely rare, we have found only 4 reported cases including our case in the world literature and only 2 cases in the Japanese literature.

Female↗
PubMed1991

Changes in immunoreactivity for cathepsin H in rat type II alveolar epithelial cells and its proteolytic activity in bronchoalveolar lavage fluid over 24 hours.

Variations in immunoreactivity for cathepsin H in rat type II alveolar epithelial (type II) cells and in its proteolytic activity in bronchoalveolar lavage fluid (BALF) were examined at six evenly spaced times over 24 hr (light period: 0600-1800 hr). Ring-shaped immunodeposits for cathepsin H were detected in type II cells (lamellar bodies), their sizes varying over 24 hr. The large ring-shaped immunodeposits increased during the light period, decreasing rapidly from 2000 to 0000 hr. Small intense immunodeposits abundantly appeared in the cells at 0000 hr. The area densities of immunodeposits in type II cells and their optical densities also varied with the time of day; both densities were high during the dark period, peaking at the mid dark period, whereas they were decreased during the light period. Proteolytic activities of cathepsin H in BALF (6 ml/rat) examined at each time revealed a distinct variation over 24 hr, corresponding to the variation in the immunoreactivity in type II cells. The activities in BALF were high from 1600 to 0400 hr and low at 0800 hr. These results suggest that the variation in immunoreactivity for cathepsin H in type II cells over 24 hr reflects the intracellular growth of lamellar bodies and secretory activity of the cells. Similar variations in the immunoreactivity and proteolytic activity of cathepsin H in the cells and BALF indicate its cosecretion with surfactants.

Animals↗
PubMed1991

Immunocytochemical localization of cathepsins B, H, and L in the rat gastro-duodenal mucosa.

Cathepsins B, H, and L are representative cysteine proteinases in lysosomes of a large variety of cells. Previous immunochemical studies indicated the presence of these enzymes also in the gastrointestinal wall. Using specific antisera, the cellular and subcellular distribution of cathepsins B, H, and L in rat gastric (oxyntic and pyloric part) and duodenal mucosa was investigated by light and electron microscopical immunocytochemistry. The subtypes of cathepsins were distributed differently in the cellular constituents of the epithelia: Cathepsin B was localized to lysosomes of all cells except goblet cells. Cathepsin H was found predominantly in gastric parietal cells (lysosomes) and in secretion granules of pyloric gastrin and duodenal cholecystokinin cells. Cathepsin L immunoreactivities were weak and restricted to a minority of cells (gastric mucous cells, enterocytes). Interstitial cells of the lamina propria immunoreactive for cathepsins H and L were identified as macrophages. The present findings suggest a dual function of cathepsins in the gastro-duodenal mucosa. They (1) cleave enzymatically proteins and peptides ingested in lysosomes, and (2) they may be involved in the processing of biologically active peptides (enteric hormones) from their precursor proteins.

Animals↗
PubMed1991

Topology of chromogranin A and secretogranin II in the rat anterior pituitary: potential marker proteins for distinct secretory pathways in gonadotrophs.

Chromogranins (Cg)/secretogranins (Sg) are representative acidic glycoproteins in secretory granules of many endocrine cells where they are co-stored and co-released with resident amines or peptides. The exact distribution of these proteins in the rat anterior pituitary is unknown. Therefore, pituitaries from untreated male rats were investigated by light- and electron-microscopical immunocytochemistry for the cellular and subcellular localization of CgA, CgB, and SgII. Endocrine cells, identified light-microscopically as gonadotrophs in adjacent semithin sections immunostained for follicle-stimulating hormone (FSH) and luteinizing hormone (LH), concomitantly were immunoreactive for CgA, CgB, and SgII. Ultrastructurally, gonadotrophs exhibited two types of secretory granules which varied in their immunoreactivities for gonadotropins and Cg/Sg. Large-sized (500 nm), moderately electron-dense granules showed antigenicities for FSH, LH, and CgA. Smaller-sized (200 nm), electron-dense granules were immunoreactive exclusively for LH and SgII. The distinct localization of CgA and SgII to morphologically and hormonally different secretory granules indicates the existence of two regulated secretory pathways in rat pituitary gonadotrophs. Hence, these proteins are considered as valuable tools to analyze the intracellular trafficking during granule biogenesis and the possible different regulation of FSH and LH secretion.

Animals↗
PubMed1991

Cysteine proteinases in bronchoalveolar epithelial cells and lavage fluid of rat lung.

We examined the presence of cathepsins B, H, and L in bronchoalveolar epithelial cells, including alveolar macrophages, and in bronchoalveolar lavage fluid (BALF), using immunocytochemistry and immunoblotting. By light and electron microscopy, immunoreactivity for cathepsins B, H, and L was detected in lysosomes of ciliated and non-ciliated epithelial cells of bronchi and bronchioles, and in macrophages. Immunodeposits for cathepsin H only were demonstrated in lamellar bodies of Type II alveolar epithelial cells, suggesting the cosecretion of surfactants and cathepsin H from the cells into the alveolar space. By immunoblotting, cathepsins B and H were found to be present in BALF. To further investigate the origin of these enzymes in BALF, alveolar macrophages obtained from BALF were cultured for 6 hr in a serum-free medium. Immunoblotting revealed that protein bands corresponding to the pro-form and mature form of cathepsin B and the mature form of cathepsin H were present in the culture medium. From these results, the presence of cathepsins B and H in BALF can be explained by the fact that cathepsin B is secreted from alveolar macrophages and cathepsin H is secreted mainly with surfactants from Type II cells and also from macrophages.

Animals↗
PubMed1991

Immunocytochemical localization of cathepsin B in rat anterior pituitary endocrine cells, with special reference to its co-localization with renin and prorenin in gonadotrophs.

We examined by immunocytochemistry the localization of cathepsin B in endocrine cells of rat anterior pituitary lobe, using a monospecific antibody to cathepsin B. By light microscopy, granular immunodeposits for cathepsin B were detected in most endocrine cells of anterior pituitary lobe. Cells immunoreactive for luteinizing hormone (LH) were diffusely immunostained by anti-cathepsin B. By electron microscopy, immunogold particles for cathepsin B were localized in lysosomes of thyrotrophs, somatotrophs, and mammotrophs. In mammotrophs, immunogold particles for cathepsin B were also detected in crinophagic bodies. Double immunostaining co-localized immunogold particles for LH and cathepsin B in secretory granules of gonadotrophs. Immunocytochemistry was also applied to demonstrate localization of renin and prorenin in LH-producing gonadotrophs; immunogold particles for renin were co-localized with those for LH, cathepsin B, or prorenin in their secretory granules. Immunogold particles for prorenin were also co-localized with those for LH or cathepsin B in secretory granules, but prorenin-positive granules appeared less frequently than renin-positive granules. These results suggest that cathepsin B not only plays a role in the protein degradation in lysosomes of anterior pituitary endocrine cells but also participates in the activation of renin in gonadotrophs, as has been demonstrated in secretory granules of juxtaglomerular cells.

Animals↗
PubMed1991

[Proliferative activity assessed on the basis of DNA ploidy patterns in primary lung cancer].

Flow cytometric cellular DNA-RNA content analysis by acridine orange staining was conducted on surgical fresh specimens of primary lung carcinomas from 66 patients (31 squamous cell carcinomas, 34 adenocarcinomas and 1 large cell carcinoma). The frequency of aneuploid tumors was 84.6% among the tumors. RNA content (RNA Index) in the DNA aneuploid tumor much more significantly (p less than 0.05) increased than the DNA diploid tumor. Tumor doubling time in the DNA aneuploid tumor was significantly (p less than 0.05) shorter than in the DNA diploid tumor. In the patients with lung cancers that recurred within 1 years, recurrence of the DNA aneuploid tumor was higher than the DNA diploid tumor. It is evident from the above results that proliferative activity in the DNA aneuploid tumor increases much more than in the DNA diploid tumor. This in turn may induce early recurrence in patients with lung cancer.

Adenocarcinoma↗
PubMed1991

[Analysis of flow cytometric nuclear DNA content of the thymoma and its relationship to malignant intensity].

Determination was made of the nuclear DNA content of paraffin-embedded specimens of resected thymoma using flow cytometry in 39 patients. Relationships among nuclear DNA content, clinicopathological findings and prognosis were studied. The frequency of DNA aneuploid tumors was 30.8%, 6.7% in stage I (15 patients), 28.6% in stage II (7 patients), 44% in stage III (9 patients) and 62.5% in stage IV (8 patients) according to Masaoka's classification, indicating more in increase with advancing stage and significant (p less than 0.05) more increase invasive thymoma than in noninvasive thymoma. The 5 year and 10 year survival rates of DNA diploid tumors were each 94%, while DNA aneuploid tumors, 75% and 45%. Patients with DNA aneuploid tumors showed less favorable prognosis than those with DNA diploid tumors. Similar results were found in patients with invasive thymoma and in those on whom subtotal or partial resection were performed. The present results indicate DNA aneuploid tumors to have greater malignant intensity than DNA diploid tumors in resected thymoma. Flow cytometric nuclear DNA content analysis provides useful biological data and new indices for evaluating the malignancy of resected thymoma.

Adult↗
PubMed1990

Effect of metabolic alterations on the density and the contents of cathepsins B, H and L of lysosomes in rat macrophages.

Crude lysosomal preparations from non-cultured peritoneal rat macrophages were shown to separate into high-density fractions rich in cathepsin B and H and low-density fractions rich in cathepsin L when layered on Percoll density gradients. Morphologically, the heavy lysosome fractions were found to consist mainly of lysosomes labeled with gold particles for anti-(cathepsin B, H and L). The light lysosome fractions contained lysosomes labeled with anti-(cathepsin B, H and L) and many other contaminants. In addition, small vesicles labeled by anti-(cathepsin L) were detected in these fractions. Addition of calf serum to the cultured macrophages induced an increase in the density of lysosomes in both dose-dependent and time-dependent fashions. Cathepsins B, H and L all shifted to the heavy lysosome fractions following the addition of serum. Progressive increase in fluorescence-labeled calf IgG in the heavy lysosome fractions after its addition suggests that the continuous entrance of excess proteins to lysosomes causes an increase in their density. This idea is supported by the fact that the density of lysosomes increased in parallel with the accumulation of horseradish peroxidase taken up in the heavy lysosome fractions. Increase in the density of lysosomes after treatment with ethyl(2S,3S)-3[(S)-3-methyl-1-(3-methyl-butylcarbamoyl)]oxirane-2- carboxylate (E-64-d) was marked in the cells cultured with serum-containing medium but slight in serum-deprived cells. However, the level of pyruvate kinase, an autophagic sequestration marker in heavy autolysosomes from E-64-d-treated cells, was much higher in serum-deprived cells, indicating that the contribution of heterophagic sequestration towards an increase in the density of lysosomes is much greater than that of autophagy.

Animals↗