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Biomedical subjects

M Matsuda

Publications and source records attributed to M Matsuda.

At least 307 records · Page 17Linked to original sources

Intergranular bridges in the anterior pituitary cell and their possible involvement in Ca2+-induced granule-granule fusion.

Quick-freeze deep-etch electron microscopy showed the presence of bridge-like structures between adjacent secretory granules in rat anterior pituitary secretory cells. These intergranular bridges were variable in length and thickness. The finest bridges were 7-8 nm in length, while the longest ones were as long as 80 nm. Annexin II, one of the Ca2+-dependent phospholipid-binding proteins, is known to interlink between two membranes and induce aggregation of liposomes and chromaffin granules under the presence of Ca2+. In anterior pituitary cells, annexin II was detected by immunoelectron microscopy at the contact sites of secretory granules with other granules. The anterior pituitary cells treated under the presence of extracellular Ca2+ with Clostridium perfringens enterotoxin which induces Ca2+ influx showed multigranular exocytosis, i.e., multiple fusions of secretory granules with each other and with the plasma membrane. The granule-granule fusion in progress could be captured by the quick-freeze deep-etch technique. The membranes of adjacent secretory granules were partially fused at their contact sites where intergranular strands were no longer seen, while there existed intergranular strands between unfused portions of the granule membranes. From these results, we consider that the intergranular bridges, some of which may be composed of annexin II, are involved in Ca2+-induced granule-granule fusion in anterior pituitary cells.

Animals↗

Usefulness of hepatitis C virus RNA counts by second generation HCV bDNA-probe in chronic hepatitis C based on the HCV genotype.

Detection of hepatitis C virus (HCV) RNA by a second generation (ver 2) HCV bDNA-probe method (bDNA-probe) was compared with detection by the first generation (ver 1) assay. The two assays were performed simultaneously with the same serum samples of HCV genotypes 1b, 2a, 2b, 3a, and 3b. The positive rates with ver 1 were 82% for HCV genotype 1b (type 1b), 57.6% for HCV genotype 2a (type 2a), 75.0% for HCV genotype 2b (type 2b), 55.6% for HCV genotype 3a (type 3a), and 93.8% for HCV genotype 3b (type 3b). The positive rates with ver 2 were 95.0% for type 1b, 93.9% for type 2a, 83.3% for type 2b, 100% for type 3a, and 93.8% for type 3b. With Fisher's exact test, the detection rate for type 2a was significantly higher (P = 0.001) with ver 2 than with ver 1. We obtained regression lines using the HCV counts measured by bDNA-probe on the y axis and the HCV counts obtained by an HCV reverse transcriptase (RT)-competitive polymerase chain reaction method (competitive PCR) on the x axis. The gradients for types 1b, 2a, and 3b were greater with ver 2 compared to ver 1. The gradients for types 2a and 3b were the highest: for type 2a, y = 0.135x + 0.6 with ver 1 and y = 0.248x + 0.1 with ver 2; for type 3b, y = 0.366x + 0.1 with ver 1 and y = 0.727x + 0.3 for ver 2. In addition, HCV-RNA counts for all the genotypes tested in this study were significantly higher with ver 2 than with ver 1. Hence, we conclude that ver 2 of the bDNA-probe measures HCV-RNA counts closer to those obtained with competitive PCR than the ver 1 assay.

DNA Probes↗

Metastatic malignant meningioma of the liver with hypoglycemia: report of a case.

We herein present the case of a 68-year-old male who suffered an episode of hypoglycemic shock 2 years after undergoing total removal of a bifrontal parasagittal malignant meningioma. Imaging studies revealed three giant hypervascular tumors with a cystic portion in the right lobe, but no confirmed preoperative diagnosis could be made. At laparotomy, liver tumors were found in the medial segment of the left lobe as well as in the right lobe, and thus an extended right lobectomy was performed. All the resected tumors were histologically diagnosed as metastatic malignant meningiomas of the liver. Despite subsequent transarterial chemoembolization for a recurrence in the residual liver, the patient died 11 months after surgery. To the best of our knowledge, only one other case of a hepatectomy for liver metastases from an intracranial malignant meningioma has been reported in the literature, but there has never been any report of surgical treatment for a metastatic meningeal tumor in the liver associated with hypoglycemia. Although our surgical treatment provided effective palliation, the prognostic significance of a surgical strategy for such patients has yet to be established.

Aged↗

Human monocyte-endothelial cell interaction induces platelet-derived growth factor expression.

OBJECTIVE: The purpose of this study was to investigate whether the synthesis of platelet-derived growth factor (PDGF), a major mitogen and chemoattractant for vascular smooth muscle cells, was induced by the direct cell-to-cell interaction between human monocytes and umbilical vein endothelial cells (ECs). METHODS: PDGF protein and mRNA expression were determined by cellular ELISA, immunohistochemical and Northern blot analyses. RESULTS: Coculture of monocytes and ECs secreted a large amount of PDGF into the supernatant, whereas culture of ECs or monocytes alone induced low levels of PDGF production. In Northern blot analysis, substantial amounts of PDGF-A and -B mRNA were induced by coculture of monocytes with ECs. Immunohistochemistry revealed that PDGF-B chain protein was detectable in both ECs and monocytes. PDGF production by ECs induced by conditioned medium of the coculture was significantly inhibited by Abs against interleukin-1 beta (IL-1 beta) and tumor necrosis factor- alpha (TNF alpha). CONCLUSIONS: These results indicate that the direct cell-to-cell interaction between human monocytes and ECs induces PDGF synthesis in both types of cells, suggesting that PDGF produced locally by monocyte-EC adhesive interaction plays an important role in the pathogenesis of atherosclerosis by promoting the migration and accumulation of vascular smooth muscle cells.

Antibodies, Monoclonal↗

Expression of prolactin messenger ribonucleic acid in the mouse gonads during sexual maturation.

The pituitary hormone prolactin is known to be produced in various extrapituitary tissues as well. We examined the presence of prolactin mRNA in the mouse gonads by reverse transcription (RT)-PCR and nucleotide sequence analysis. Both ovary and testis were found to express small amounts of mRNA coding prolactin, identical to that in the pituitary gland. Next, we established a sensitive competitive RT-PCR method to estimate the amount of prolactin mRNA and then measured its expression in the gonads during sexual maturation (10-80 days of age). In the ovary, the amount of prolactin mRNA (copies/microg of total RNA) gradually decreased from day 10 to 40, and the lower level was maintained until day 80. Little difference in the amount was observed between the estrous and diestrous groups on day 80. Conversely, the amount of testicular prolactin mRNA gradually increased from day 10 to 80. These results suggest that prolactin is produced in the mouse gonads and that it acts as an autocrine/paracrine factor modulating gonadal functions.

Animals↗

Crystallization, fluoridation and some properties of apatite thin films prepared through rf-sputtering from CaO-P2O5 glasses.

Using calcium phosphate glass targets with the CaO/P2O5 molar ratios of 1.50-0.50, much lower than the stoichiometric value of 3.3 for hydroxyapatite, thin films of stoichiometric hydroxy-, nonstoichiometric oxyhydroxy- and Ca-deficient oxyhydroxy-apatites were prepared on alumina ceramic substrates by rf-sputtering followed by post-annealing. Based on the present results, a phase diagram for CaO-P2O5 at low temperatures in the ambience of air was depicted for thin films. The ambient H2O vapor had an influence on the phase diagram: Tricalcium phosphate was changed to apatite in the presence of H2O vapor. Dense fluorohydroxyapatite thin films were prepared by fluoridation of those apatite thin films at a low temperature such as 200 degrees C. In the present report, some functional properties of thin films thus prepared were also shown.

Aluminum Oxide↗

Two types of brain chondroitin sulfate proteoglycan: their distribution and possible functions in the rat embryo.

The distribution of neurocan-like and 6B4 proteoglycan-like immunoreactivities in the rat embryo was investigated from gestational days 10.5-15.5 with monoclonal antibody 1G2 or 6B4 that immunoreacted with neurocan and 6B4 proteoglycan, respectively. In the brain region, the leptomeningeal layer in the myelencephalon, metencephalon, diencephalon or telencephalon was first stained with monoclonal antibody 1G2 at embryonic day 12.5. In the spinal cord, monoclonal antibody 1G2 stained the regions corresponding to the boundary caps (designated the boundary caps) after embryonic day 11.5 and the roof plate after embryonic day 12.5. The intensity of staining in the boundary caps reached a maximum at embryonic day 13.5, at around the time when the axons from the dorsal root ganglia reach this region. However, the points of contact of the axons with the boundary caps were hardly stained. By contrast, the roof plate was most strongly and widely stained at embryonic day 14.5, at around the time when the axons enter the spinal cord. Western blotting of preparations from the spinal cord that included the boundary caps revealed the presence of neurocan in this region. Thus, it is likely that neurocan serves as a barrier molecule to regulate the direction of axonal growth from the dorsal root ganglia. By contrast, in addition to staining of the future brain and spinal cord, monoclonal antibody 6B4 stained the trigeminal and sympathetic ganglia in the rat embryo on and after embryonic day 12.5, as well as the vestibular, facial and dorsal root ganglia after embryonic day 12.5. In studies in tissue culture, monoclonal antibody 6B4 prevented the inhibitory effects of 6B4 proteoglycan on the proliferation of PC12D cells. No immunostaining with monoclonal antibody 6B4 was observed in cells that had incorporated bromodeoxyuridine in vivo. Possible functions of 6B4 proteoglycan in the rat embryo are discussed.

Animals↗

Microbial contamination of 'sterile water' used in Japanese hospitals.

We examined the following samples of water from 10 hospitals for microbial contamination: water obtained using an ultra filtration system (UF water), a reverse osmosis system (RO water), a water distillation system (distilled water) and tap water. UF water and RO water are used for handwashing before surgery, and distilled water for the preparation of drugs. All 10 samples of tap water examined were contaminated with < 10 colony forming units (cfu)/mL. Thirteen (68%) of 19 samples of UF water, nine (53%) of 17 samples of RO water and 15 (79%) of 19 samples of distilled water were contaminated with 10(1)-10(4) cfu/mL. The majority of micro-organisms were non-fermentative bacteria such as Sphingomonas paucimobilis and CDC gr. IV C-2. Japanese hospitals commonly use UF water and RO water for preoperative handwashing under the assumption that it is sterile. Our results suggest, however, that these types of water are inferior microbiologically to tap water. Distilled water from the dispensary was also contaminated with micro-organisms. The available chlorine content of tap water was 0.17-0.42 ppm and that of UF water (from tap water) 0-0.06 ppm. There was no available chlorine in RO water or distilled water (each from tap water). The reduction or disappearance of available chlorine appears to be associated with microbial contamination of UF water, RO water and distilled water.

Chlorine↗

Constitutive association of EGF receptor with the CrkII-23 mutant that inhibits transformation of NRK cells by EGF and TGF-beta.

Crk belongs to the adapter proteins that participate in many signalling pathways from cell surface receptors. We have characterised the CrkII-23 mutant that inhibits the transformation of NRK cells induced by epidermal growth factor (EGF) and transforming growth factor (TGF)-beta. To study the biochemical difference, cDNAs of the wild-type CrkII and the CrkII-23 mutant were introduced stably into NIH 3T3 cells expressing EGF receptor (EGFR). Both CrkII and CrkII-23 were phosphorylated on tyrosine upon EGF simulation with similar time course and dose dependency. Whereas the wild-type CrkII bound to EGFR only after EGF stimulation, CrkII-23 bound to EGFR from before stimulation. Mutation in the Src homology (SH) 2 or amino-terminal SH3 domain did not abolish the binding of CrkII-23 to EGFR in the quiescent cells, suggesting that the binding is mediated by a novel mechanism. These CrkII-23-derived mutants, however, did not suppress transformation of NRK cells by EGF and TGF-beta. Hence, both the SH2 and amino-terminal SH3 domains are required to inhibit transformation of NRK cells. These results suggest that persistent signalling from CrkII-23 bound to EGFR suppresses transformation by EGF and TGF-beta in NRK23 cells.

3T3 Cells↗

Molecular genotyping by pulsed-field gel electrophoresis of restricted genomic DNA of strains of Taylorella equigenitalis isolated in Ireland and in the United States.

The profiles, after digestion with ApaI or NotI and pulsed-field gel electrophoresis (PFGE), of genomic DNA from 18 strains of Taylorella equigenitalis isolated in Ireland were of three different types. The 13 strains in one of these types gave PFGE profiles identical to that of an American prototype strain, Kentucky 188, but different from strain NCTC11184T and from a strain isolated in Japan. Eight additional strains isolated in the United States gave four distinct types of genomic PFGE profiles. All four types were different from that of T. equigenitalis NCTC11184T or that of the Japanese strain. The profile of three strains of one type was identical to that of Kentucky 188.

Animals↗

Gastrointestinal amyloidosis secondary to hypersensitivity vasculitis presenting with intestinal pseudoobstruction.

A 22-year-old woman developed sudden hepatic encephalopathy and severe intestinal bleeding. She was diagnosed with acute fatty liver and hypersensitivity vasculitis and was successfully treated with whole plasma exchange, methylprednisolone pulse therapy, and transcatheter arterial embolization. Twenty-seven months later, she began complaining of lower abdominal fullness, tenderness, and nausea and vomiting. Histologic examination showed that she had developed gastrointestinal and renal amyloidosis with intestinal pseudoobstruction and proteinuria. The immunohistochemical study of the stomach, rectum, and kidney with anti-amyloid A fluorescent antibody showed that the systemic amyloid deposit was secondary to her underlying disease. This is the first report of amyloidosis occurring secondary to hypersensitivity vasculitis.

Adult↗

Decreased expression of signal-transducing CD3 zeta chains in T cells from the joints and peripheral blood of rheumatoid arthritis patients.

Although T cells from patients with rheumatoid arthritis (RA) have previously been determined to have poor proliferative responses to a variety of stimuli, the underlying mechanism is not known. We have investigated the expression of the signal-transducing zeta molecule in subsets of T cells and natural killer (NK) cells derived from the peripheral blood mononuclear cells (PBMC) and synovial fluid mononuclear cells (SFMC) of RA patients using quantitative flow cytometry, Western blot analysis and immunohistochemistry. A decrease of zeta expression was apparent in all investigated lymphocyte subsets from the PBMC and SFMC of RA patients, as compared to the corresponding subsets from healthy age- and sex-matched controls. A less pronounced reduction of cell surface-located CD3 epsilon, CD4 and CD8 was also located in T cells from SFMC as compared to PBMC from RA patients. Biochemical demonstration of the low or absent CD3 zeta in PBMC from patients with RA was achieved by Western blot analysis. Immunohistochemical staining and image analysis also confirmed the low expression of zeta chains in synovial tissue of RA patients. The possibility that the decreased expression of zeta and of immune functions of T cells from RA patients may be related to the presence of free oxygen radicals, as we have previously reported in cancer patients, should be considered.

Arthritis, Rheumatoid↗

Fluorine-18 fluorodeoxyglucose positron emission tomography imaging of parotid mass lesions.

Using X-ray CT or magnetic resonance imaging (MRI), fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) studies were performed in 28 patients with parotid lesions. All lesions except two showed higher accumulation of FDG than normal parotid gland. High accumulation was found in all of 5 carcinomas, all of 6 Warthin's tumors, and 8 of 12 pleomorphic adenomas, with standardized uptake values (SUVs) over 3.0. The mean SUVs of carcinomas, Warthin's tumors, and pleomorphic adenomas were 5.92 +/- 2.05, 7.03 +/- 2.49, and 4.07 +/- 1.55, respectively. On the other hand, all 5 inflammatory lesions demonstrated low accumulation, with mean SUVs of 1.66 +/- 0.89. It is thus difficult to differentiate malignant from benign parotid lesions by SUV on FDG-PET.

Adult↗

Role of the kringle domain in plasminogen activation with staphylokinase.

We have evaluated the effect of lysine binding sites in kringle structures on the activation of plasminogen with plasmin and staphylokinase (SAK) complex and on the binding of plasminogen to SAK. Activation of native plasminogen (Glu-plasminogen) by a catalytic amount of plasmin-SAK complex increased in the presence of epsilon-amino-n-caproic acid (EACA) and then decreased with higher concentrations of EACA. By contrast, activation of modified plasminogen (Lys-plasminogen) decreased in an EACA-concentration-dependent manner. This decrease was explained by a more than 10-fold higher Km for activation of Lys-plasminogen with a catalytic amount of plasmin-SAK complex in the presence of EACA. EACA was a competitive inhibitor with Ki 0.23 mM. In addition, the Km for activation of mini-plasminogen, which lacks first four kringle structures (K1+2+3+4), was at least 3.5-fold higher than that for the activation of Lys-plasminogen. Furthermore, EACA showed a negligible inhibitory effect on the activation of mini-plasminogen by the plasmin-SAK complex. We observed a similar biphasic effect of EACA on the binding of Glu-plasminogen to SAK and a dose-dependent effect on the Lys-plasminogen binding to SAK by gel filtration methods. Since EACA binds to plasminogen via lysine binding sites in the kringle structure, we propose that the lysine binding site in K1+2+3+4 domain plays a role in the activation of plasminogen by plasmin SAK complex, and in the binding of plasminogen to SAK.

Aminocaproic Acid↗

Opposing effects of low and high molecular weight kininogens on cell adhesion.

High molecular weight kininogen (HK) blocks cell spreading but not cell attachment to surfaces coated with vitronectin and other ligands of beta3 integrins. We sought to learn the structural basis of this phenomenon. Monoclonal antibodies against the histidine-rich D5 domain in the light chain of 2-chain HK abolished the inhibitory effect of 2-chain HK on spreading of MG-63 osteosarcoma cells on vitronectin-coated tissue-culture plastic. The antibodies were effective only if incubated with 2-chain HK in solution and did not abolish the anti-cell-spreading effect of 2-chain HK that was pre-adsorbed to tissue-culture plastic. Exposure of an epitope in the histidine-rich domain was less when HK was adsorbed to tissue-culture plastic (oxidized polystyrene) than when it was adsorbed to ELISA plastic (untreated polystyrene). Loss of the epitope correlated with increased anti-cell-spreading activity of HK on tissue-culture plastic. The light chain of 2-chain HK containing D5 and that containing recombinant D5 both had anti-cell-spreading activity, but only when present in solution during adhesion assays. Pre-adsorption of recombinant D5 to tissue-culture plastic resulted in a surface on which adsorbed 2-chain HK had little anti-cell-spreading activity. Binding study revealed that HKa bound to immobilized vitronectin. The histidine-rich D5 domain of light chain of HK was identified as one of the binding sites of vitronectin, suggesting that the masking of the RGD cell-binding site of immobilized vitronectin is the molecular mechanism of anti-cell-spreading effect of HKa. In contrast, low molecular weight kininogen (LK), which lacks D5, augmented cell spreading on vitronectin-coated tissue-culture plastic. Thus, HK and LK have opposing effects on VN-dependent cell adhesion. The augmenting effect of LK was greater if LK was preincubated with cells or adsorbed to the surface at pH>7.0. Analysis of fragments of LK and antibody inhibition studies localized the cell-adhesion activity to the D3 domain that is common to LK and HK. These findings indicate that the D5 domain mediates the adsorption of HK or 2-chain HK to vitronectin substratum in anti-adhesive conformations, i.e., masking of the RGD cell-binding site of vitronectin. Such conformers inhibit cell spreading on vitronectin even though a cell-adhesion site is present in D3.

Antibodies, Monoclonal↗

Gastric small-cell carcinoma in Japan: a case report and review of the literature.

The authors encountered a patient with primary gastric small-cell carcinoma (primary gastric SCC). Histologic examination revealed combined carcinoma containing non-small-cell elements such as adenocarcinoma and squamous cell differentiation. On preoperative biopsy, only well-differentiated adenocarcinoma was collected. Since gastric SCC was first reported in 1976, this type of carcinoma has been detected in 54 patients in Japan. Concerning these cases, the authors reviewed the light and/or electron microscopic findings of neuroendocrine granules, positive reaction for neuron-specific enolase staining, and the histologic type of resected specimens. There were only three patients positive for neuron-specific enolase in which light and electron microscopy demonstrated neuroendocrine granules. However, 50 patients (approximately 93%) overall satisfied at least one of these findings. Concerning the histologic characteristics of the resected specimens, multidirectional differentiation was more markedly observed compared with that in SCC of the lung. Adenocarcinomatous and/or squamous differentiation was observed in more than half the cases, and preoperative biopsy suggested carcinoma other than SCC or undifferentiated carcinoma in several cases.

Aged↗

Endothelin-1 in the heart during exercise.

In addition to its potent vasocontractile effects, endothelin-1 (ET-1) has potent positive inotropic and chronotropic effects on isolated heart muscles in vitro. However, it is not known whether the production of ET-1 in the heart is altered by exercise. In this study we investigated the production of ET-1 in the heart during exercise. Rats performed treadmill running of 45-min duration. Sedentary rats served as controls. Immediately after exercise, the heart was quickly removed. The peptide level of ET-1 in the heart was measured by a sandwich-enzyme immunoassay. The peptide level of ET-1 in the heart was significantly higher in the exercise group than in the control sedentary group. Therefore, we have demonstrated that production of ET-1 in the heart is increased by exercise. The present study suggests that myocardial ET-1 participates in the modulation of cardiac function during exercise.

Animals↗