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Biomedical subjects

Y Yamaguchi

Publications and source records attributed to Y Yamaguchi.

At least 595 records · Page 33Linked to original sources

K-glypican: a novel GPI-anchored heparan sulfate proteoglycan that is highly expressed in developing brain and kidney.

Glypicans are a family of glycosylphosphatidylinositol (GPI)-anchored cell surface heparan sulfate proteoglycans (HSPGs). The glypican family, which currently includes glypican, the developmentally regulated rat intestinal transcript OCI-5, and cerebroglycan, is characterized by a similar core protein size and almost complete conservation of cysteine residues. By RT-PCR using degenerate oligonucleotide primers based on the sequence homologies, we isolated mouse cDNA encoding a novel member of the glypican family as well as mouse homologues of glypican and OCI-5. The novel molecule, named K-glypican, has a predicted molecular mass of 57.5 kD and potential attachment sites for heparan sulfate chains and a GPI anchor in its COOH-terminal region, like other members of the glypican family. Transfection of an epitope-tagged full-length K-glypican cDNA into MDCK cells demonstrated that K-glypican is indeed expressed as a GPI-anchored HSPG. Northern blot analyses with K-glypican, glypican, and OCI-5 probes demonstrated that kidney and developing brain, and that these three molecules show remarkable patterns of cell type- and developmental stage-specific expression. In situ hybridization revealed that the major sites of K-glypican expression in developing embryo are tubular epithelial cells in the kidney and proliferating neuroepithelial cells in the brain. These results indicate that K-glypican is a novel GPI-anchored HSPG involved in embryonic development.

Amino Acid Sequence↗

Lysine-49-phospholipases A2 from Trimeresurus flavoviridis venom are membrane-acting enzymes.

Basic proteins I and II (BP-I and BP-II) isolated from the venom of Trimeresurus flavoviridis (Habu snake) are isozymes of highly active Asp-49 phospholipase A2 (Asp-49-PLA2) and classified into the group Lys-49-PLA2. BP-II was found to elicit a strong contraction of guinea pig ileum, and this activity was inhibited completely by 1 microM indomethacin, an inhibitor of the arachidonate cascade. BP-II was inactive in the Ca(2+)-free medium, and p-bromphenacylated His-48-BP-II was also inactive. BP-II exhibited no binding affinity for the cells expressing PLA2 receptors. These results indicated that the contraction elicited by BP-II is due to the hydrolytic action of BP-II, liberating arachidonic acid from the ileum phospholipid biomembranes. In spite of its limited lipolytic activities (av. 0.9% of Asp-49-PLA2) against monomers and micelles of synthetic phospholipids, BP-II hydrolyzed considerably strongly the phospholipids in the artificial bilayer vesicles. Arachidonic acid released from liposomes of beta-arachidonoyl-gamma-stearoyl-L-alpha-phosphatidylcholine was determined by HPLC, and the activity of BP-II was estimated to be about 75% as compared to Asp-49-PLA2. Liposomes encapsulating carboxyfluorescein exhibited a strong dye-leakage induced by BP-II in a concentration-dependent manner, only in the Ca(2+)-containing buffer. The net result from all these observations was that BP-II, a Lys-49-PLA2, is an enzyme that hydrolyzes the membrane phospholipids. In contrast to BP-II, BP-I was found to be considerably weak in hydrolyzing membrane phospholipids, although its activities were distinct. BP-I and BP-II share a common sequence with the sole exception of Asp-67 (BP-I) and Asn-67 (BP-II) in the aligned sequences. This implies that the amino acid at position 67 of Lys-49-PLA2s is the residue required for discriminatory recognition of beta-arachidonoyl-phospholipid membranes.

Animals↗

Polysialogangliosides expressed by amelanotic melanoma: a possible explanation for the poor response to anti-monosialoganglioside antibody 202 in a patient with melanoma.

A 52-year-old Japanese woman developed numerous amelanotic metastatic melanomas on the skin and in various organs three years after a surgical operation for primary melanoma on the right axilla. The patient was treated with monosialoganglioside specific monoclonal antibody 202; however, no apparent clinical effects were observed. Ganglioside analysis of a metastatic tumor demonstrated that it expressed GM3, GM2, GD3, GD2, and polysialogangliosides. Since polysialogangliosides rarely appear in melanomas, their expression may explain the patient's poor response to MAb 202. The relationship between ganglioside composition and the effect of anti-ganglioside monoclonal antibody is discussed.

Antibodies, Monoclonal↗

Different cholesterol deposition in aorta of Dahl salt-sensitive rats and spontaneously hypertensive rats fed a high-cholesterol diet.

1. We compared the serum and aortic lipid levels in spontaneously hypertensive rats (SHR) and Dahl salt-sensitive rats (DSR) fed a high-cholesterol (HC) diet. 2. In SHR fed the HC diet, the serum cholesterol level significantly increased, but no aortic cholesterol deposition was observed. 3. The serum cholesterol level in DSR fed the HC diet markedly increased compared to that in DSR fed the basal diet, and this change was greater with the diet containing 8% NaCl than 0.4% NaCl. A significant increase in the content of aortic cholesterol, notably cholesteryl ester, was observed in only DSR fed the HC diet containing 8% NaCl. 4. These results suggest that the combination of hypercholesterolaemia with salt-induced hypertension acts as a greater risk factor for atherosclerosis than that with genetic hypertension.

Animals↗

Comparison of the effects of hypercholesterolaemia on relaxation responses in aortas of spontaneously hypertensive rats and Dahl salt-sensitive rats.

1. We investigated the effects of hypercholesterolaemia on relaxation responses in thoracic aortas isolated from two different types of hypertensive rats; spontaneously hypertensive rats (SHR) and Dahl salt-sensitive rats (DSR). 2. All rats fed the high cholesterol diet for 8 weeks showed a significant increase in the serum cholesterol level. The high cholesterol diet did not change the blood pressure of SHR, but increased that of hypertensive DSR fed a high-salt diet. 3. In aortas of SHR, the high-cholesterol diet did not change the endothelium-dependent and -independent relaxation induced by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively. 4. In aortas of hypertensive DSR, the high-cholesterol diet notably reduced the ACh-induced relaxations and slightly reduced SNP-induced relaxation. 5. These results suggest that hypercholesterolaemia causes greater impairment of endothelium-dependent relaxation in rat aorta with salt-induced hypertension than genetic hypertension.

Acetylcholine↗

Suppression of sympathetic nervous system is involved in hypotension and bradycardia during hemofiltration in anesthetized dogs.

Hypotension occurring during hemodialysis is often accompanied by paradoxical bradycardia. However, the mechanism is poorly understood. This study was designed to determine the role of the sympathetic nervous system in hemodialysis-induced hypotension and bradycardia. We measured efferent sympathetic nerve activities to the heart (CNA), kidney (RNA), liver (HNA), spleen (SpNA), and adrenal gland (AdNA), along with heart rate (HR), blood pressure (BP), central venous pressure (CVP), and left atrial pressure (LAP) during hemofiltration performed at a rate of 0.3 mL .center dot Kg(-1) center dot min(-1) for 30 min in anesthetized dogs. The response to hemorrhage was also studied at the same bleeding speed. Hemorrhage caused a decrease in BP (-18 +/- 1 mmHg; 1 mmHg = 133.3 Pa) with reflex increases in HR (7 +/- 2 beats/min) and sympathetic nerve activities. In contrast, hemofiltration caused a decrease in CNA (85 +/- 18%), HNA (86 +/- 11%), and SpNA (88 +/- 11%) with greater decreases in BP (-43 +/- 10 mmHg) and HR (-27 +/- 14 beats/min) than hemorrhage. During hemofiltration, the decreases in BP, HR, CNA, HNA, and SpNA were attenuated after vagotomy. Hematocrit increased by 6.5% at 30 min after hemofiltration, whereas it decreased by 4.3% after hemorrhage. These results suggest that hemofiltration suppresses the sympathetic nervous system, resulting in decreases in HR and BP. Furthermore, this sympathetic suppression during hemofiltration is mediated by vagal afferents.

Animals↗

Mitosis and the presence of binucleate cells among glomerular podocytes in diseased human kidneys.

Podocytes are highly differentiated cells that appear to be postmitotic. However, it is unclear whether cell division or the proliferation of podocytes takes place in the setting of glomerular injury. In this study, we analyzed the presence of mitotic figures and of binucleate or multinucleate podocytes in diseased human kidneys by light and electron microscopy. Among the 164 cases analyzed, mitotic figures in podocytes were found in only one cell, in a case of focal and segmental glomerulosclerosis, and 6 cases provided evidence of two or more nuclei in a single podocyte. Serial sections by light microscopy confirmed that the nuclei in multinucleate cells were separate and of approximately the same size. Electron-microscopic analysis of the binucleate podocytes revealed hypertrophy and effacement of foot processes. These findings indicate that, in human podocytes, mitosis without cytokinesis (cell division) results in binucleate cells in cases of glomerular injury.

Biopsy↗

Immunological study of anti-M2 in antimitochondrial antibody-negative primary biliary cirrhosis.

We investigated the specificity of various autoantibodies in antimitochondrial antibody (AMA)-negative patients with primary biliary cirrhosis (PBC). We examined sera from 144 patients with PBC, 17 of whom were AMA negative by indirect immunofluorescence. The AMA-negative group showed a significantly higher positivity for smooth muscle antibody, but not for antinuclear antibody, as compared with the AMA-positive group. IgG class anti-PDH by enzyme-linked immunosorbent assay (ELISA) were detected in 13% of the AMA-negative group. However, all PBC patients showed positive IgG, IgA, and/or IgM class anti-M2 to the four M2 proteins by immunoblotting. These results suggest that the detection of IgG and IgM class anti-PDH and that of antibodies to the four M2 proteins increases the positivity of this ELISA method, and that detection of IgG, IgA, and IgM class anti-M2 to the four M2 proteins by immunoblotting is useful in diagnosing AMA-negative patients with PBC.

Adult↗

[A case of male infertility with a chromosomal abnormality of 45, X/46, X + mar].

A 28-year-old man visited our hospital with a complaint of male infertility. His development was normal with body size of 169.5 cm in height and muscular type. His beard and axillary and pubic hairs were male type. The penis and scrotum were also normal, but testes were small (8 ml, bilateral). Semen analysis revealed azoospermia. Serum luteinizing hormone (LH) and testosterone levels are within normal range but follicle stimulating hormone (FSH) was moderately elevated. Testicular biopsy specimen showed severely hyalinized seminiferous tubuli, incarceration of basement membrane and hyperplasia of Leydig cells. Johnsen's score count was two. Chromosomal analysis revealed a mosaic type of 45, X/46, X + mar. We tried to confirm the existence of sex-determining region of the Y chromosome (SRY) by the DNA analysis. It was shown that SRY on genomic DNA fragment. This marker chromosome may be translocated to a fragment of the short arm of Y chromosome.

Adult↗

Amidepsines, inhibitors of diacylglycerol acyltransferase produced by Humicola sp. FO-2942. I. Production, isolation and biological properties.

Humicola sp. FO-2942, a soil isolate, was found to produce a series of new inhibitors of diacylglycerol acyltransferase (DGAT). Three active compounds, designated amidepsines A, B and C, were isolated from the fermentation broth of the producing strain by solvent extraction, silica gel column chromatography, ODS column chromatography and HPLC. Amidepsines inhibit DGAT activity with IC50 values of 10.2 approximately 51.6 microM in an enzyme assay system using rat liver microsomes. Amidepsines also showed specific inhibition of triacylglycerol formation in intact Raji cells, indicating that they inhibit DGAT activity in living cells.

Acyltransferases↗

Endoscopic therapy for early gastric cancer. Utility of endosonography and evaluation of prognosis.

We analyzed 30 endoscopically treated patients with early gastric cancer in terms of the utility of endoscopic ultrasonography (EUS) and the evaluation of prognosis. Endoscopic therapy for early gastric cancer included strip biopsy, laser irradiation, and OK-432 injection. Examination of the depth of cancer invasion in the strip biopsy specimens showed that the diagnostic accuracy of EUS was 85.7%. Complete resection rate in strip biopsy was 82.8%, and the patients with incompletely resected cancer were successfully treated with laser irradiation and/or OK432 injection. The prognosis of the patients treated with strip biopsy was good. Three patients died of causes other than gastric cancer. Local recurrence and a heterochronous new lesion developed in six patients, but all of these cases were successfully treated with additional strip biopsy. All but one patient treated with laser irradiation and/or OK 432 injection survived during the study period. These results suggest that EUS is useful in determining the indication of endoscopic therapies and that prognosis is good in patients with early gastric cancer treated endoscopically.

Aged↗

[The structure and function of the renal interstitial cells].

The renal interstitial cells consist of fibroblast-like cells, lipid-laden interstitial cells, macrophages, interstitial dendritic cells and perivascular cells. The fibroblast-like cells are mainly located in renal cortex and medulla, and have long cytoplasmic processes forming a reticular network between the tubules and capillaries. In disease state, interstitial cells show expressions of HLA-class 2 antigen, ICAM-1 and VCAM-1, and the expressions are related with leukocyte infiltrate and tubulitis. In tubular injury and obstruction, the proliferation of interstitial cells occurs around damaged tubules, and form a lymphatic system. In the interstitial fibrosis, interstitial and tubular epithelial cells secrete procollagen and interstitial cells have the action of resolving them. Therefore, the relationship between interstitial fibrosis and interstitial cells is not yet clear. The medullary interstitial cells have receptors for angiotensin 2 or bradykinin and secrete PGE2. The interstitial cells show an interaction of blood components and tubular epithelial cells.

Animals↗

[T cell receptor V beta repertoire of locoregional lymphocytes in malignant effusions].

We analyzed the T cell receptor (TCR) V beta repertoire of lymphocytes obtained from patients with malignant effusion treated by locoregional immunotherapy. Polymerase chain reaction using a panel of V beta subfamily specific oligonucleotide primers(V beta 1-20) was employed after reverse transcription of mRNA isolated from the locoregional cells. No major oligoclonality of TCR repertoire was observed in effusion lymphocytes before the treatment. The expression level of V beta 13.1 repertoire was significantly higher in effusion lymphocytes than in peripheral blood lymphocytes before treatment. V beta 20 gene expression increased significantly after locoregional administration of OK-432. It is suggested that TCR V beta 13.1 may be responsible for effusion lymphocytes and TCR V beta 20 for OK-432-related antigenic stimulus.

Adult↗

[Mutation of the K-ras oncogene in pancreatic carcinoma, and application of its detection in pancreatic juice to diagnose pancreatic carcinoma].

The incidence of the point mutation of K-ras oncogene at codon 12 in surgical or autopsy specimens of pancreatic carcinoma (PC) is reportedly extremely high ranging from 75 to over 90%, and the K-ras mutation found in PC is almost exclusively present in codon 12. The GGT to GAT transition of the 12th codon is observed in more than 50% of PC of Japanese patients, although the incidences of transition of GGT to CGT, GTT, and GAT are essentially equal in PC of European countries. With this point in mind, the authors attempted to detect K-ras mutations in DNA obtained from pancreatic juice (PJ) collected endoscopically. K-ras mutations at codon 12 were found in 55% of 20 PC patients by PCR and ASO probe hybridization method, and in 80% of 25 PC patients by PCR-RFLP method. On the other hand, K-ras mutations were negative in PJ from chronic pancreatitis with one exception by PCR-RFLP method. Other authors also reported almost the same results. These results suggest that analysis of K-ras oncogene in PJ can be useful for qualitative diagnosis of PC, and would be expected as a novel tool for early detection of PC.

DNA, Neoplasm↗

[Correlation between acute rejection and the role of donor specific cytotoxic T lymphocyte in canine lung allotransplantation].

We investigated the correlation between the donor specific CTL activities and pathological findings of the grafted lungs by using mononuclear cells obtained from BAL fluids (BAL), peripheral blood (PBL), spleen (SpL) and grafted lungs (GIL) in canine lung allotransplantation. Twenty eight mongrel dogs underwent left lung allotransplantation. Fifteen dogs were not given immunosuppressive drugs. Thirteen dogs were given cyclosporine a 20 mg/kg/day per os for ten days postoperatively. Dogs were monitored by chest roentgenograms and cytotoxic activity tests of which targets were donor's fibroblasts. Pathological findings were classified by Prop's classification. The results were as follows. In latent and vascular phases no elevations of cytotoxic activities were demonstrated in PBL, BAL and GIL. But in early alveolar phase the cytotoxic activity was elevated to 19.83 +/- 14.54% in BAL. In late alveolar phase the highest cytotoxicities of 30.83 +/- 15.28% in PBL, 72.63 +/- 7.36% in BAL and 70.07 +/- 6.34% in GIL at E:T = 100:1 were demonstrated. In pneumonia cytotoxicity did not elevate at all in any effectors. As a result of them, it clearly shows that BAL is a good tool for the diagnosis of lung rejection.

Animals↗