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Biomedical subjects

M Matsuda

Publications and source records attributed to M Matsuda.

At least 397 records · Page 22Linked to original sources
PubMed1997

Fibrin detected in plasma of patients with disseminated intravascular coagulation by fibrin-specific antibodies consists primarily of high molecular weight factor XIIIa-crosslinked and plasmin-modified complexes partially containing fibrinopeptide A.

Pooled plasma from 40 patients with severe disseminated intravascular coagulation (DIC) secondary to septic conditions was subjected to gel permeation chromatography on Sephacryl S-500 HR after sample pretreatment with KSCN for dissociation of non-covalent fibrin complexes. Fibrin antigen in eluates was detected by an array of ELISA tests, using two monoclonal antibodies against fibrin degradation product D-dimer, a monoclonal antibody against an epitope generated by plasmin cleavage of the D-domain, and an antibody against the neo-N-terminus of the alpha-chain of fibrin exposed by cleavage of fibrinopeptide A. Tag antibodies were a polyclonal antibody against the fibrinogen/ fibrin D-domain, a POD-conjugated version of the monoclonal antibody against fibrin alpha-chain neo-N-terminus, and a polyclonal antibody against fibrinopeptide A. Most fibrin-related material present in the pooled DIC plasma was of higher molecular mass than fibrinogen. Fibrin polymers were reactive with antibodies against D-dimer, plasmin cleaved D-domain, and fibrin alpha-chain neo-N-terminus. Part of the polymers reacted with antibodies against fibrinopeptide A, indicating presence of fibrinogen or desA-fibrin monomer within the covalently linked complex. In conclusion, the primary analytes detected by monoclonal antibodies for D-dimer, plasmin-specific epitopes of fibrin degradation products, as well as sites exposed by fibrinopeptide cleavage in plasma from patients with disseminated intravascular coagulation are high molecular weight factor XIIIa-crosslinked fibrin complexes, containing plasmin-cleaved D-domains, intact fibrin monomer units, and fibrinogen or desA-fibrin monomer.

Antibodies, Monoclonal↗
PubMed1997

[Activation of Rap1, antagonist to ras, by Crk-C3G].

Rap1 was identified as gene whose overexpression suppressed transformation by ras. Rap1 belongs to the Ras family. The amino acid sequences of Rap1 and Ras show 55% identity to each other. Due to this high sequence similarity, Rap1 binds to effector molecules of Ras, however, Rap1 does not activate them. Thus, Rap1 functions are antagonistic to Ras in the cells. C3G was identified as a Crk SH3-binding guanine nucleotide exchange factor. Biochemical and cell biological analyses revealed that C3G is a Rap1 activator. Since it has been considered that Crk transduces signals from tyrosine kinases, this finding suggests that the activity of Rap1 is also under the control of tyrosine kinases. Overexpression of C3G in ras-transformed cells caused the morphology of the cells to revert to that of normal cells. Moreover, a mutant cell line that was resistant to EGF-dependent transformation was isolated. In the cell line a mutation was found in crk gene that was the cause of the resistance. These findings suggest that Crk-C3G-Rap1 pathway may function as an anti-transformation machinery.

Animals↗
PubMed1997

[Antinociceptive activity of intracisternal clonidine in the mouse].

The antinociceptive activities of clonidine have been determined against three qualitatively different noxious stimuli in the mouse. The methods used to evaluate this activity were selected to include tests which employ different types of noxious stimuli, i.e. heat (hot plate), chemical (acetic acid-induced writhing) and mechanical (tail pinch). Test drug and control treatments were given by cisternal injection in a dose volume of 10 microliters.mouse-1. The results presented here show that clonidine has potent antinociceptive properties against several types of noxious stimuli. Clonidine produced steep dose-response lines in all tests. The response to the writhing assays were completely inhibited by 1.0 microgram.mouse-1 of clonidine. In contrast in both the hot plate and tail pinch assay, however, clonidine did not produce a consistent antinociceptive effect at a dose of 200 micrograms.mouse-1. Utilizing these three different types of assays, the rank order of antinociceptive potency for clonidine in different noxia was the writhing >> hot plate > tail pinch. It was concluded from these results that clonidine has potent antinociceptive properties against chemical visceral stimuli.

Adrenergic alpha-Agonists↗
PubMed1997

[Mitochondrial redox change in gerbil hippocampus before and after transient ischemia].

To investigate the basis of neuronal vulnerability we studied mitochondrial redox changes in gerbil hippocampus before and after 5 minutes forebrain ischemia. The brain was frozen by in situ funnel freezing method, and grinned off coronally until exposure of hippocampus. Relative value of regional redox ratio (NAD+/NADH) was obtained from fluorescence signals of intrinsic fluorochromes, i.e., NADH (PN) and flavoproteins (Fp), using a high resolution fluorometer. We calculated a modified redox ratio MRR = FP/(Fp + PN). Each point is displayed in gray scales ranged 16 degrees corresponding to the MRR value of the point; black represents a low MRR value (reduced) and white represents a high value (oxidized). Pyramidal cell layers and the granule cell layers were seen as linear areas of high MRR. The stratum radiatum and stratum orience of the CA 1 subfield showed low MRR compared with other hippocampal regions. During ischemic period, MRR in all subfield of hippocampus had decreased but the decrease was more severe in CA 1 region than in another. Just after recirculation, MRR decreased transiently in dentate and CA 3 areas but was fully recovered in all hippocampal areas with the exception of CA 1 region, where the MRR decreased again 12 hours after recirculation. These results suggest that CA 1 area suffers more pronounced hyoxic condition (state V) than other less vulnerable regions during 5 minutes ischemia. The irreversible reduction of MRR in CA 1 area may result from continuing mitochondrial dysfunction, and this may cause lasting energy shortage in CA 1 neurons that eventually results in slowly progressive cell death.

Animals↗
PubMed1997

Random amplified polymorphic DNA observed in Eucalyptus by PCR study with random primers.

A total DNA extracted from Eucalyptus globlus, E. citriodora, or a subspecies of E. gunnii (Maruha Yu-kari) being used as an template, Polymerase Chain Reaction (PCR) was performed with 40 kinds of commercially available random primers (Operon, OPA-01-20 and OPB-01-20). After electrophoresis of PCR-products on agarose gel, we analyzed the band pattern formed by DNA-fragments with 0.5-3.0 kbp as molecular weight. As a remuneration for the examination in which forty 10-mer arbitrary oligonucleotides were tried to be used as primers in PCR for each species, we found that 9 primers (for E. globlus) leads polymorphism, 6 primers for E. citriodora, and 6 primers for Maruha Yu-kari.

Base Sequence↗
PubMed1997

Novel primers designed for microsatellite loci in Eucalyptus and identification by PCR fingerprints.

We found a novel PCR-primer which can be used for the identification on "elite-tree-selection". This primer was designed for selective hybridization at the both ends of microsatellite loci, which is well known as one of the most highpervariable region of DNA. After PCR-fingerprinting on five Eucalyptus species (E. globulus, E.citriodora, E.grandis, E. maidenii, E.bicostata), with our primer, DNA-polymorphism was observed all over the cases.

Base Sequence↗
PubMed1997

Identification of Eucalyptus citriodora clones micropropagated in tissue culture.

The extent of genetic identity observed in the young individuals which were micropropagated from a single Eucalyptus individual was analyzed by using DNA-fingerprinting. Among 40,000 tissue-cultured-seedings of E.citriodora, 200 plants were randomly chosen so that each total DNA might be extracted from their leaves. Using these DNAs as template, PCR was performed with some primers we found in advance that leads polymorphism for DNA of E. citriodora. In this study, all over the 200 cases, the band pattern formed cDNA fragment on a gel after electrophoresis was the identical one mutually.

Base Sequence↗
PubMed1996

Occult Breast Cancer: A Case Report and Review of the Literature.

We report a case of a 44-yar-old woman with occult breast cancer presenting as an axillary mass in whom sonography was able to detecct an involved internal mammary node also, thus helping to establish a diagnosis of breast cancer. The patient underwent extended radical mastectomy, including internal mammary lymphadenectomy. Microscopy of the removed specimen failed to find a primary breast cancer lesion. Metastatic cancer was seen in the palpable axillary node, another resected axillary node and a removed internal mammary node. The estrogen and progesterone receptor analysis of the axillary node were negative. Since occult breast cancer was found highly potential for metastasizing to the infraclavicular or internal mammary nodes, it is reasonable to treat such patients in the same way as those with palpable breast cancer, with adjuvant chemotherapy.

Journal Article↗
PubMed1996

B cell antigen receptor signaling induces the formation of complexes containing the Crk adapter proteins.

Crk proteins are Src homology (SH) 2/SH3-containing adapter proteins that can mediate the formation of signaling complexes. We show that engaging the B cell antigen receptor (BCR) on the RAMOS B cell line caused both Crk-L and Crk II to associate with several tyrosine-phosphorylated proteins. We identified two of these phosphoproteins as Cas and Cbl and showed that both bound to the Crk SH2 domain after BCR engagement. BCR ligation also increased the amount of Crk proteins in the particulate fraction of the cells and induced the formation of Crk.Cas and Crk.Cbl complexes in the particulate fraction. We propose that tyrosine phosphorylation of membrane-associated Cas and Cbl creates binding sites for the Crk SH2 domain and recruits Crk complexes to cellular membranes. Thus, Crk proteins may participate in BCR signaling by using their SH2 domains to direct the interactions and subcellular localization of proteins that bind to their SH3 domains. In RAMOS cells, we found that the SH3 domains of Crk-L and Crk II bound C3G. Since C3G activates Rap, a negative regulator of the Ras pathway, Crk proteins may participate in regulation of Ras signaling by the BCR.

Adaptor Proteins, Signal Transducing↗
PubMed1996

CrkII signals from epidermal growth factor receptor to Ras.

A rat fibroblast mutant defective in oncogenic transformation and signaling from epidermal growth factor receptor to Ras has been isolated. The mutant contains dominant negative-type point mutations in the C-terminal SH3 domain of one crkII gene. Among the adapters tested, the mutant is complemented only by crkII cDNA. Expression of the mutated crkII in parent cells generates the phenotype indistinguishable from the mutant cell. Yet overexpression or reduced expression of Grb2 in the mutant before and after complementation with crkII have little effect on its phenotype. We conclude that adapter molecules are highly specific and that the oncogenic growth signal from epidermal growth factor receptor to Ras is predominantly mediated by CrkII in rat fibroblast.

Alleles↗
PubMed1996

A monoclonal antibody that recognizes a neo-antigen exposed in the E domain of fibrin monomer complexed with fibrinogen or its derivatives: its application to the measurement of soluble fibrin in plasma.

Using urea-solubilized human fibrin monomer as an immunogen, we raised in mice a battery of monoclonal antibodies that reacted with the immunogen but not with urea-treated or native fibrinogen. Although they all failed to react with acid-solubilized fibrin monomer (acid-FM) alone, an antibody designated as IF-43 was found to recognize acid-FM, which was bound with fibrinogen or its derivatives to form a 1:2 complex of soluble fibrin. The epitope for this antibody, thus, appears to be exposed most probably by conformation changes induced in the acid-FM molecule upon formation of the complex. Because IF-43 was able to recognize fibrin-derived plasmic fragment E treated with urea but not the thrombin- and urea-treated amino-terminal disulfide knot of fibrinogen, the presence of the A alpha (52-78) residue segment seems to be prerequiste for the epitope expression. The antibody was found to react with soluble fibrin monomer spiked to normal plasma dose-dependently up to 200 micrograms/mL. By an aggregation assay using latex beads coated with IF-43, we found that concentrations of soluble fibrin monomer in plasma derived from patients with thrombotic diseases were mostly elevated, but not necessarily correlated with those of the D-dimer, reflecting another aspects of the disease. Furthermore, the soluble fibrin monomer in plasma derived from patients with thrombotic diseases was found to be depleted solely of the A peptides, but not the B peptides, based on its subunit polypeptide compositions lacking the beta-chain on immunoblotting.

Animals↗
PubMed1996

Interaction between the amino-terminal SH3 domain of CRK and its natural target proteins.

CRK is a human homolog of chichen v-Crk, which is an adaptor protein. The SH2 domain of CRK binds to several tyrosine-phosphorylated proteins, including the epidermal growth factor receptor, p130(Cas), Shc, and paxillin. The SH3 domain, in turn, binds to cytosolic proteins of 135-145, 160, 180, and 220 kDa. We screened expression libraries by Far Western blotting, using CRK SH3 as a probe, and identified partial cDNA sequences of four distinct proteins, including C3G, DOCK180, EPS15, and clone ST12. The consensus sequence of the CRK SH3 binding sites as deduced from their amino acid sequences was Pro+3-Pro+2-X+1-Leu0-Pro-1-X-2-Lys-3. The interaction of the CRK SH3 domain with the DOCK180 peptide was examined with an optical biosensor, based on the principles of surface plasmon resonance. A low dissociation constant of the order of 10(-7) resulted from a high association rate constant (kassoc = 3 x 10(4)) and low dissociation rate constant (kdiss = 3 x 10(-3)). All CRK-binding proteins except clone ST12 also bound to another adaptor protein, Grb2. Mutational analysis revealed that glycine at position +1 of ST12 inhibited the binding to Grb2 while retaining the high affinity binding to CRK SH3. The result suggests that the amino acid at position +1 also contributes to the high affinity binding of the peptides to the SH3 domain of Grb2, but not to that of CRK.

Adaptor Proteins, Signal Transducing↗