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Biomedical subjects

N Aoki

Publications and source records attributed to N Aoki.

At least 631 records · Page 35Linked to original sources
PubMed1982

Cross-linking of alpha 2-plasmin inhibitor to fibrin catalyzed by activated fibrin-stabilizing factor.

During blood coagulation alpha 2-plasmin inhibitor (alpha 2PI) is cross-linked with fibrin by an activated fibrin-stabilizing factor (FSFa) plasma transglutaminase, activated coagulation factor XIII). When alpha 2PI was treated with FSFa in the absence of acceptor amino groups, the inhibitor lost more than 90% of its capacity to be cross-linked to fibrin because of hydrolysis of the gamma-carboxamides of FSFa-susceptible glutamine residues. Chemical modifications of the inhibitor's lysine epsilon-amino groups did not affect the cross-linking capacity of the inhibitor with fibrin, whereas the same chemical modifications in fibrinogen resulted in a remarkable loss of cross-linking capacity. These observations suggest that alpha 2PI plays a role as an acyl donor with its FSFa-susceptible glutamine residues in the cross-linking reaction with fibrin, and fibrin serves as an acyl acceptor with its lysine residues. The number of FSFa-susceptible glutamine residues/molecule of the inhibitor was estimated by measuring the maximum incorporation of [3H]histamine into the inhibitor and by analyzing the distribution of radioactivity in a tryptic digest of [14C]histamine-incorporated alpha 2PI.l It was found that each inhibitor molecule has one glutamine residue that is most susceptible to FSFa. When the radioactive histamine-incorporated inhibitor was reacted with excess amounts of plasmin, a small fragment carrying all the released radioactivity was rapidly released from the NH2-terminal part of the inhibitor moiety of the complex. The NH2-terminal amino acid sequence of the inhibitor was analyzed before and after treatment with FSFa or before and after incorporation of radioactive histamine. The glutamine residue at the second position from the NH2-terminal end was converted to a glutamic acid residue when the inhibitor was treated with FSFa. When the radioactive histamine-incorporated inhibitor ws analyzed, the radioactivity was found predominantly at the second position from the NH2-terminal end. These results indicate that the glutamine residue susceptible to FSFa in alpha 2PI is located next to the NH2-terminal residue.

Acyltransferases↗
PubMed1982

Reversible cross-linking of alpha 2-plasmin inhibitor to fibrinogen by fibrin-stabilizing factor.

alpha 2-Plasmin inhibitor, a primary inhibitor of fibrinolysis, is cross-linked to fibrin by plasma transglutaminase (glutaminyl-peptide:amine gamma-glutamyltransferase, EC 2.3.2.13, activated fibrin-stabilizing factor) when blood coagulation takes place. alpha 2-Plasmin inhibitor was found also to be cross-linked to fibrinogen by plasma transglutaminase. The inhibitor was corss-linked exclusively to the A alpha-chain of fibrinogen, and the cross-linking reaction proceeded very rapidly. The reaction was almost completed before the formation of the gamma-chain dimers of fibrinogen which precedes cross-linking polymerization of the A alpha-chain of fibrinogen. The maximum level of inhibitor cross-linking achieved was approx. 30% of the inhibitor present at the start of the reaction. The level of cross-linking of the inhibitor was not changed when the cross-linking reaction was preceded by dimerization of fibrinogen. The cross-linking reaction was found to be a reversible one, since the cross-linked complex of the inhibitor and fibrinogen was partly dissociated to each of its components when the complex was incubated with plasma transglutaminase. These results suggest that the self-limiting nature of the cross-linking reaction between alpha 2-plasmin inhibitor and fibrin(ogen) is due to the reaction equilibrium favoring dissociation of the complex, and not due to the development of structural hindrance in polymerizing fibrin(ogen).

Acyltransferases↗
PubMed1982

Plasminogen Tochigi: inactive plasmin resulting from replacement of alanine-600 by threonine in the active site.

Structural studies on a hereditarily abnormal plasminogen, plasminogen Tochigi, have been performed to identify the difference responsible for its lack of proteolytic activity. The plasminogen sample used was from a heterozygote and thus consisted of apparently equal amounts of normal and defective plasminogen molecules. Amino acid sequence analysis of a tryptic peptide isolated from the abnormal plasminogen indicated that Ala-600 (equivalent to Ala-55 in the chymotrypsin numbering system) had been replaced by Thr. No other substitutions in the active-site residues--namely, His-57, Asp-102, and Ser-195--were found. Molecular models for chymotrypsin and the bovine trypsin-pancreatic trypsin inhibitor complex indicate that Ala-55 is very near the active-site His. The Thr at position 55 in plasminogen (plasmin) Tochigi may perturb His-57 such that the proton transfers associated with the normal catalytic process cannot occur in the abnormal plasmin.

Alanine↗
PubMed1982

Mucoepidermoid carcinoma of the thymic region.

A case of mucoepidermoid carcinoma in thymus in a 59-year-old Japanese female is presented. She died of cardiac tamponade due to tumor invasion ater a 5 years' clinical course. At autopsy the main tumor was found in the thymic region with metastases to the sternum, regional lymph nodes, pericardial, and left pleural cavity. The mucoepidermoid carcinoma might be probably originated from a hen's egg-sized cyst which was located in the upper posterior aspect of the tumor-involved thymus. No teratomatous components were present. The cyst was most likely to be of thymic or bronchogenic cyst origin, though it was not determined, in view of the lining with pseudostratified ciliated columnar epithelium of the cystic wall and the surrounding with the thymic tissue outside. Moreover, there was thymic hyperplasia with germinal center that was compatible with SLE-like symptoms in her past history and autoimmune nature of the autopsy findings of pulmonary fibrosis.

Carcinoma↗
PubMed1982

Antipsychotic effects of caerulein, a decapeptide chemically related to cholecystokinin octapeptide, on schizophrenia.

Caerulein, a decapeptide chemically related to CCK-8, was administered intramuscularly to 20 patients with chronic schizophrenia in two different doses of 0.3 and 0.6 microgram/kg. BPRS ratings were made before and 3 weeks after the injection. The neuroleptic therapy was not discontinued, but both drug and dose were not changed at least 3 weeks before the first injection and during the study period. Clinically obvious and statistically significant improvement in psychotic symptoms occurred shortly after the injection of caerulein. The greatest change occurred 1--2 weeks later. There was an evident correlation between the observed changes and the dose injected. Our findings suggest that caerulein has a long-acting, antipsychotic activity in chronic schizophrenia. Furthermore, our findings suggest the involvement of CCK-like peptides in the pathogenesis of schizophrenia.

Adult↗
PubMed1982

Significance of cross-linking of alpha 2-plasmin inhibitor to fibrin in inhibition of fibrinolysis and in hemostasis.

When blood is clotted, alpha(2)-plasmin inhibitor (alpha(2)PI) is cross-linked to fibrin by activated fibrin-stabilizing factor (activated coagulation Factor XIII, plasma transglutaminase). The amount of cross-linked alpha(2)-PI is proportional to the amount of alpha(2)PI present at the time of clotting. Plasma from a patient with congenital deficiency of alpha(2)PI was supplemented with various amounts of purified alpha(2)PI. Clots were prepared from these plasmas and were suspended in plasma containing a normal concentration of alpha(2)PI, and spontaneous clot lysis was observed. When the clot was formed in the presence of calcium ions and thereby allowing cross-linking to occur, the rate and extent of fibrinolysis were found to be inversely proportional to the concentrations of alpha(2)PI present in the clot at the time of clotting. When the clot was formed in the absence of calcium ions so that no cross-linking occurred, the clot underwent fibrinolysis at similar rates, regardless of the concentrations of alpha(2)PI in the clot. When the clot formed in the presence of calcium ions was squeezed and washed to remove unbound proteins before being suspended in plasma, the extent of fibrinolysis was also inversely proportional to the amount of alpha(2)PI cross-linked to fibrin. Similar results were obtained when the clot was suspended in buffered saline instead of plasma. These observations suggest that spontaneous fibrinolysis is mainly carried out by plasminogen/plasminogen activator bound to fibrin, and this fibrinolysis caused by fibrin-associated activation of plasminogen was mainly inhibited by alpha(2)PI cross-linked to fibrin. To further support this concept, alpha(2)PI treated with activated fibrin-stabilizing factor and that had lost most of its cross-linking capacity was used in similar experiments. This modified alpha(2)PI had the same inhibitory activity on plasmin as the native inhibitor, but gave significantly less inhibition of fibrinolysis in every experiment, particularly when the clot was compacted by platelet-mediated clot retraction or by squeezing. Thus, it was concluded that alpha(2)PI cross-linked to fibrin plays a significant role in inhibition of physiologically occurring fibrinolysis. It is further suggested that the absence of cross-linked alpha(2)PI contributes to accelerated fibrinolysis and hemorrhagic tendency in patients with congenital deficiency of fibrin-stabilizing factor.

Calcium↗
PubMed1982

Mutagenic/carcinogenic potential of DEHP and MEHP.

The mutagenic/carcinogenic activities of DEHP and MEHP were studied in bacteria and mammalian cells. MEHP but not DEHP exerted a dose-dependent DNA damaging effect to B. subtilis in Rec-assay. DEHP and MEHP showed mutagenic activities to S. typhimurium TA-100, with and without S-9 mix, respectively. MEHP produced not only the mutation in E. coli WP2B/r but also sister chromatid exchanges (SCE) in Chinese hamster V79 cells. It also induced 8AG/6TG-resistant gene mutations and chromosomal aberrations in the V79 cells. Transplacental administration of DEHP or MEHP to the Syrian golden hamster embryos was carried out by administering DEHP or MEHP to gravid animals on day 11 of gestation, followed by the cultivation of embryonic cells for 15-20 days. Both DEHP and MEHP induced 8AG/6TG-resistant mutation, chromosomal aberrations and morphological transformation in the embryonic cells of the Syrian golden hamster.

Animals↗
PubMed1982

Ultrastructural analysis of hepatitis B surface antigen production in vitro.

Recently several continuous cell lines (among these PLC/PRF/5 cells) producing hepatitis B surface antigen (HBsAg) were established from human hepatocellular carcinomas. The cultured cells provide the first opportunity to study HBsAg synthesis and secretion in vitro. HBsAg, but not HBcAg, was localized by the fluorescent antibody technique in the cytoplasm and on the surface of the cultured cells. Under the electron microscope, th PLC/PRF/5 cells displayed morphologic characteristics of both hepatocytes and hepatocellular carcinoma cells. However, 22-nm. spherical or filamentous HBsAg particles were not seen in the cells, although spherical HBsAg particles were observed in the supernatant culture media. Therefore, the indirect immunoperoxidase technique was used to demonstrate HBsAg at the ultrastructural level. Electron-dense reaction product was detected along the nuclear envelope, on rough-surfaced endoplasmic reticulum, and in cisternae of endoplasmic reticulum. These findings suggest that HBsAg is synthesized on rough-surfaced endoplasmic reticulum and transferred into endoplasmic cisternae for processing and secretion. This mode of HBsAg production is identical with that observed in hepatocytes of patients infected with hepatitis B virus. The absence of detectable intracellular HBsAg particles suggests that the cultured cells secrete the particles very rapidly or that they may have a defect in intracisternal packaging of HBsAg into particles.

Adult↗