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Biomedical subjects

M Irie

Publications and source records attributed to M Irie.

At least 73 records · Page 4Linked to original sources

Amino acids and peptides. LIV. Application of 2-adamantyl derivatives as protecting groups to the synthesis of peptide fragments related to Sulfolobus solifataricus ribonuclease. I.

The 2-adamantyloxycarbonyl group was employed for the protection of the epsilon-amino group of Lys and the hydroxyl group of Tyr, and the 2-adamantyl ester was employed for the protection of the beta-carboxyl group of Asp in order to construct eight peptide segments as building blocks for the preparation of peptide fragments related to the sequence of Sulfolobus solifataricus Ribonuclease. The usefulness of the above protecting groups developed in our laboratory was confirmed.

Adamantane↗

Amino acids and peptides. LV. Application of 2-adamantyl derivatives as protecting groups to the synthesis of peptide fragments related to Sulfolobus solifataricus ribonuclease. II.

Segment condensations were performed to construct peptide fragments related to Sulfolobus solifataricus Ribonuclease. At each condensation step, the new protecting groups were stable. The protected peptide fragments were treated with a low-high HF procedure to give the desired peptide fragments. These peptide fragments were also prepared by the solid-phase method, and the obtained peptides were compared with those obtained by the solution method. The peptide fragments obtained by the solution method were identical with those obtained by the solid-phase method on analytical HPLC, indicating that the new protecting groups could be easily removed by HF, and no racemization occurred during the synthesis of the protected peptides.

Adamantane↗

Comparison of base specificity and other enzymatic properties of two protozoan ribonucleases from Physarum polycephalum and Dictyostelium discoideum.

Base specificity and other enzymatic properties of two protozoan RNases, RNase Phyb from a true slime mold (Physarum polycephalum) and RNase DdI from a cellular slime mold (Dictyostelium discoideum), were compared. These two RNases have high amino acid sequence similarity (83 amino acid residues, 46%). The base specificities of two base recognition sites, The B1 site (base recognition site for the base at 5'-side of scissile phosphodiester bond) and the B2 site (base recognition site for the base at 3'-side of the scissile bond) of the both enzymes were estimated by the rates of hydrolysis of 16 dinucleoside phosphates. The base specificities estimated of B1 and B2 sites of RNase Phyb and RNase DdI were A, G, U > C and A > or = G > C > U, and A > or = G, U > C and G > U > A, C, respectively. The base specificities estimated from the depolymerization of homopolynucleotides and those from the releases of four mononucleotides upon digestion of RNA coincided well with those of the B2 sites of both enzymes. Thus, in these enzymes, the contribution of the B2 site to base specificity seems to be larger than that of the B1 site. pH-stability, optimum temperature, and temperature stability, of both enzymes are discussed considering that RNase Phyb has one disulfide bridge deleted, compared to the RNase DdI with four disulfide bridges.

Amino Acid Sequence↗

Evaluation of porcine fat with fiber-optic spectroscopy.

Spectroscopy using four types of fiberoptic probes and a sensor at wavelengths of 400 to 1,100 nm was evaluated to assess porcine fat quality. The shapes of the spectrum for the leaf fat with white color and various firmnesses differed with the type of probe: surface, contact, insertion, or transmittance. The internal reflectance ratio using the insertion probe at wavelengths from approximately 600 to 1,000 nm was positively correlated with the hardness, melting point, and saturated fatty acid content of the fat, but it was negatively correlated with the refractive index and polyunsaturated fatty acid content. The correlations between the internal reflectance ratio using the insertion probe and the monounsaturated fatty acid content were strong only near 1,100 nm. Surface reflectance at more than 650 nm was negatively correlated with refractive index. Transmittance at almost all wavelengths showed positive correlations with monounsaturated fatty acid content, but it was negatively correlated with hardness, melting point, and saturated fatty acid content. The interactance using the contact probe did not have a significant correlation with any physiochemical characteristics. The strongest relationships for hardness, refractive index, saturated fatty acid content, monounsaturated fatty acid content, polyunsaturated fatty acid content, and melting point were obtained at 650 nm (r = .88), 660 nm (r = -.91), 645 nm (r = .73), 1,095 nm (r = .68), and 930 nm (r = -.76), respectively, using the insertion probe and 1,050 nm (r = -.79) using the transmittance probe (P<.05). These results indicated that fiber-optic methods were rapid and useful techniques for the evaluation of porcine fat quality.

Adipose Tissue↗

The effect of primers on bond strength of polyacid-modified resin composites (compomers).

This study evaluated the effect of primer on shear bond strength and marginal gaps of six new compomers immediately after light-activation. A resin-modified glass ionomer cement, a conventional glass-ionomer cement and a microfilled composite were used for comparison. The marginal gap widths of each of the four compomers and a microfilled composite used with the primer were significantly smaller compared with those used without the primer. The bond strength values of five compomers used with the primer were significantly higher than those used without the primer. The bond strength of conventional glass-ionomer was not affected by the primer (or the conditioner).

Adhesiveness↗

[Fundamental and clinical study of three-dimensional compensating filters and direct dose monitoring system for total body irradiation].

We devised 3D-compensating filters to improve dose distribution during total body irradiation (TBI). This study investigated the effect of these 3D-compensating filters and related complications in patients who had undergone bone marrow transplantation (BMT) followed by TBI. The 3D-compensating filters were fabricated by CT measurement of body thickness. The effectiveness of the 3D-compensating filters in producing a homogeneous dose distribution was checked by a thermoluminescent dosimeter (TLD) and semiconductor detectors in all patients. At the pelvis, the dose was measured simultaneously with an ionization chamber. The average dose distribution to each site when the 3D-compensating filter was used was 93% to the head, 97% to the neck, 99% to the thorax, and 98% to the pelvis in TLD when the scheduled dose was taken as 100%. There was no significant difference between the TBI and non-TBI groups with regard to the frequency of lung toxicity. Clinical interstitial pneumonitis occurred in 22.7% of the patients, interstitial pneumonitis with CMV in 13.6%, and idiopathic pneumonitis in 6.1% without any virus infection. Only one patient was regarded as having radiation-induced pneumonitis. 3D-compensating filters can be conveniently produced within a short time following CT measurement, and they seem to be safe and useful for dose flattening during TBI.

Adolescent↗

BEGAIN (brain-enriched guanylate kinase-associated protein), a novel neuronal PSD-95/SAP90-binding protein.

PSD-95/SAP90 is a synaptic membrane-associated guanylate kinase with three PDZ, one SH3, and one guanylate kinase (GK) domain. PSD-95/SAP90 binds various proteins through the PDZ domains and organizes synaptic junctions. PSD-95/SAP90 also interacts with the postsynaptic density (PSD) fraction-enriched protein, named SAPAP (also called GKAP and DAP), through the GK domain. SAPAP is Triton X-100-insoluble and recruits PSD-95/SAP90 into the Triton X-100-insoluble fraction in the transfected cells, suggesting that SAPAP may fix PSD-95/SAP90 to the PSD. Here we report a novel protein interacting with the GK domain of PSD-95/SAP90, BEGAIN. BEGAIN is specifically expressed in brain and enriched in the PSD fraction. BEGAIN is Triton X-100-soluble in the transfected cells but is recruited to the Triton X-100-insoluble fraction by SAPAP when coexpressed with PSD-95/SAP90. BEGAIN may be a novel PSD component associated with the core complex of PSD-95/SAP90 and SAPAP.

Amino Acid Sequence↗

A novel multiple PDZ domain-containing molecule interacting with N-methyl-D-aspartate receptors and neuronal cell adhesion proteins.

At synaptic junctions, pre- and postsynaptic membranes are connected by cell adhesion and have distinct structures for specialized functions. The presynaptic membranes have a machinery for fast neurotransmitter release, and the postsynaptic membranes have clusters of neurotransmitter receptors. The molecular mechanism of the assembly of synaptic junctions is not yet clear. Pioneering studies identified postsynaptic density (PSD)-95/SAP90 as a prototypic synaptic scaffolding protein to maintain the structure of synaptic junctions. PSD-95/SAP90 belongs to a family of membrane-associated guanylate kinases and binds N-methyl-D-aspartate receptors, potassium channels, and neuroligins through the PDZ domains and GKAP/SAPAP/DAP through the guanylate kinase (GK) domain. We performed here a yeast two-hybrid screening for SAPAP-interacting molecules and identified a novel protein that has an inverse structure of membrane-associated guanylate kinases with an NH2-terminal GK-like domain followed by two WW and five PDZ domains. It binds SAPAP through the GK-like domain and NMDA receptors and neuroligins through the PDZ domains. We named this protein S-SCAM (synaptic scaffolding molecule) because S-SCAM may assemble receptors and cell adhesion proteins at synaptic junctions.

Adaptor Proteins, Signal Transducing↗

Interaction of rat lin-10 with brain-enriched F-actin-binding protein, neurabin-II/spinophilin.

We have recently isolated a rat homologue of the Caenorrhabditis elegans lin-10 product. Although rat lin-10 is expressed in the cytosol and membrane fractions of various tissues, it is distributed only in the membrane fraction in brain where it is enriched in the synaptic plasma membrane and postsynaptic density fractions. We have isolated here a rat lin-10-interacting protein from rat brain and identified it to be neurabin-II/spinophilin, which has recently been isolated as a protein interacting with protein phosphatase I and F-actin. Neurabin-II/spinophilin is ubiquitously expressed but enriched in brain, especially in the synaptic plasma membrane and postsynaptic density fractions. We discuss the physiological significance of the interaction of rat lin-10 with neurabin-II/spinophilin.

Actins↗

Molecular cloning and characterization of rat lin-10.

In Caenorhabditis elegans, the vulval induction is mediated by tyrosine kinase receptor/Ras signal transduction pathway composed of the lin-3, let-23, and let-60 products. In addition to these gene products, the lin-2, lin-7, and lin-10 products are also implicated in this pathway. Lin-2 encodes a MAGUK and lin-7 encodes a small protein with one PDZ domain. The lin-10 product has no homology to known proteins. Here, we have cloned a rat homologue of the lin-10 product and characterized it. Rat lin-10 is ubiquitously expressed in various rat tissues and distributed in both the cytosol and membrane fractions. In brain, however, rat lin-10 is distributed only in the membrane fraction and enriched in the synaptic plasma membrane and postsynaptic density fractions. These results suggest that rat lin-10 is involved at least in synaptic functions in brain.

Animals↗

Biochemistry of frog ribonucleases.

Frogs have unique pyrimidine base-specific RNases, with structures similar to those of turtle, iguana and chicken RNases. Among the four frog RNases discussed here, three from Rana pipiens, R. catesbeiana and R. japonica oocyte cells show antitumour activity, and the latter two show lectin activity towards sialic acid-rich glycoproteins. In this review, (i) we compare their unique primary structures with respect to the locations of their disulphide bridges, three-dimensional structure, base specificity and heat stability as compared with RNase A, and (ii) we summarize current knowledge about the mode of action of lectin and the antitumour activities of the three frog RNases.

Amino Acid Sequence↗

Changes in colonic motility in dogs after a resection of the inferior mesenteric ganglion and plexus.

In a curative resection for advanced sigmoid or rectal cancer, an extensive dissection of the regional lymph nodes is generally required. This often necessitates the removal of the autonomic nerves around the inferior mesenteric artery. The present study was done in an attempt to clarify the influence of a neurectomy around the inferior mesenteric ganglion and plexus on the motility of the colon. In eight dogs, we resected the ganglion and plexus around the inferior mesenteric artery, together with an implantation of strain gauge force transducers in various parts of the colon, and 7-10 days later, colonic motility was examined. The percentage of contractile states and contractile forces increased at both the distal colon in fasting dogs, as well as at the middle colon in the late postprandial period. At the distal colon, contractile forces were noted in the early and late postprandial periods. These contractile abnormalities at the middle and distal colon may thus explain the frequent bowel movements or diarrhea often observed after extensive surgery in patients with sigmoid or rectal cancer.

Animals↗

Role of bile acids in duodenal migrating motor complexes in dogs.

Previous studies have suggested that enterohepatic circulation of bile acids is essential for regular cycling of duodenal migrating motor complexes (MMCs). The present study was an attempt to clarify the role of bile acids in the enterohepatic circulation system in initiating duodenal MMCs. Seven dogs underwent total external biliary diversion that resulted in the loss of MMCs originating from the duodenum. Sodium ursodeoxycholate (6 mg/kg/hr) was then given either through the portal vein or a peripheral vein, and motility of the gastrointestinal tract was serially recorded. When sodium ursodeoxycholate was given either through the portal vein or a peripheral vein during external biliary diversion, duodenal MMCs restarted. The cyclic change in plasma motilin levels during an MMC cycle as induced by sodium ursodeoxycholate was almost the same as in a normal MMC cycle. Total bile acid concentration in the portal vein changed cyclically with MMC cycles when bile flow was intact but did not change cyclically with MMC cycles restarted by intravenous bile salt infusion. Bile acid stimulation of putative receptors existing between the portal vein and intrahepatic bile ducts may be involved in initiating normal duodenal MMC cycles.

Animals↗

Characterization and primary structure of a base non-specific and acid ribonuclease from Dictyostelium discoideum.

A base non-specific and acid RNase was isolated from cellular slime mold (Dictyostelium discoideum) cells in a homogeneous state (about 2.4 kDa) by SDS-polyacrylamide gel electrophoresis. The RNase (RNase DdI) has a pH optimum of 5.0. The amino acid sequence of RNase DdI was determined by a combination of protein chemistry, a search of Data base, Dicty cDB and further sequence analysis of cDNA from the same bank. RNase DdI consists of 198 amino acid residues, and about 13.3, 0.9, 1.2, 3.3, and 1.0 residues of mannose, xylose, glucose, GlcNAc, and GalNAc, respectively. RNase DdI has two characteristic conserved segments of the RNase T2 family, and thus belongs to the RNase T2 family. Considering the fact that most of the RNase activity of D. discoideum is present in the lysosomal fraction [Wiener and Ashworth (1970) Biochem. J. 118, 505-512], it was concluded that the lysosomal RNase in D. discoideum is a member of the RNase T2 family. The amino acid sequence of RNase DdI is highly homologous with that of Physarum polycephalum RNase (RNase Phyb), and its amino acid sequence seems to be similar to those of plant/animal type RNases, rather than fungal RNases. The location of RNase DdI in the phylogenetic tree of the RNase T2 family was estimated.

Amino Acid Sequence↗

A protease sensitive region of plant and animal ribonucleases belonging to the RNase T2 family.

Recombinant RNase LE from tomato and squid liver RNase Tp, typical plant/animal type RNases belonging to the RNase T2 family, were subjected to limited digestion with several proteases, and the cleavage sites were analyzed by Edman degradation. Recombinant RNase LE was cleaved specifically at the 24th Lys by lysylendopeptidase and trypsin, and RNase Tp was cleaved at the 21st Glu by V8 protease. These cleavage sites are located very close to those where the cleavage during preparation of several animal RNase T2 family enzymes was observed. From this finding, it was concluded that the short segment around the 20th amino acid residue in plant/animal RNases is located on the surface of the molecules and forms loops, and is thus very sensitive to proteases.

Amino Acid Sequence↗

Characterization of ribonucleases from culture medium of Lentinus edodes.

Lentinus edodes (shiitake) cultivated in potato dextrose medium produced five RNases in the culture filtrate. The two major RNases (RNase Le37 and RNase Le45) were highly purified and their molecular masses, base specificities, N-terminal amino acid sequences, and amino acid compositions were analyzed and compared to RNase Le2 isolated from the fruit bodies of the same mushroom. RNase Le37 and RNase Le45 are base non-specific and adenylic acid preferential RNases like RNase Le2 and their N-terminal sequences are very similar to RNase Le2, but they are glycoproteins and their amino acid compositions are significantly different from that of RNase Le2. In addition to these enzymes, a guanylic acid-specific RNase with a molecular mass 13 kDa was partially purified. Since RNase Le2, which has very similar N-terminal sequence to RNase Le37 and RNase Le45, was not excreted from the mycelia, the analysis of the structures of these two excreted RNase may shade a light on the mechanism of excretion of RNases in this organism.

Amino Acids↗

Primary structure of a squid acid and base non-specific ribonuclease.

Squid (Todarodes pacificus) liver RNase (RNase Tp) was purified. RNase Tp was a base non-specific and acid RNase. Upon hydrolysis of RNA, RNase Tp released four mononucleotides in the order of G > A > U > C. RNase Tp consisted of two peptides with 198 and 23 amino acid residues. The amino acid sequences of these peptides were analyzed. The large peptide had two unique segments containing most of the active site amino acid residues of RNase T2 family enzymes. From the comparison of the sequence of short peptide with the sequences of the other RNase belonging to RNase T2 family RNases, it was found that the amino acid sequence of the short peptide was very similar to that of the C-terminal portion of RNases of the RNase T2 family. Thus, we concluded that the short peptide was a C-terminal part of RNase Tp. The molecular mass of the protein moiety of RNase Tp was 25,582 daltons. The amino acid sequence of RNase Tp most resembles that of oyster RNase (91 amino acid residues identical) in the RNase T2 family RNases. However, the N-terminal portion of RNase Tp was unusually similar to those of plant RNases, rather than the other animal RNases.

Acids↗

Registration system for growth hormone (GH) treatment with standardized immunoreactive GH values in Japan.

The Foundation for Growth Science has been controlling the use of GH by its registration system, which includes a scoring system for the eligibility for GH treatment according to the diagnostic criteria for GH deficiency (GHD) established by the Study Group for Hypothalamo-pituitary Disorder of the Ministry of Health and Welfare. Until 1995, 28,876 patients with GHD (19,432 boys and 9,444 girls) had been registered as eligible for GH treatment. The number of patients registered in a year increased gradually till 1990 due to the unlimited hGH supply by recombinant techniques and the change in the criteria for GH treatment and the number registered became stable after 1990. The frequency of GH-treated patients is calculated to be 55.2/100,000 persons (72.2/100,000 in boys and 37.1/100,000 in girls) in patients born between 1960 and 1990. The highest frequency was 148.4/100,000 persons (191.7/ 100,000 boys and 103.7/100,000 girls) in 1981, when 2,278 patients (1,508 boys and 770 girls) were born. Eligibility for GH treatment is assessed according to the scoring system which is basically dependent on peak GH values in provocation tests so that standardization of GH values measured with the various commercial GH kits is required to avoid inequality of patients' access to the treatment. In samples obtained by GRF test in 10 normal volunteers, hGH was measured with seven human GH (hGH) kits at a laboratory center. Since the RIA value has been used historically for the diagnosis of GHD, the mean of two RIA measurements was selected as the basis for the standardization procedure and the linear regression formula was used for each hGH kit. After the freely available supply of hGH obtained by recombinant DNA techniques, the role of the Foundation for Growth Science has changed to avoid hGH abuse. Even with this regulation, the frequency of registered patients may indicate a tendency to GH overuse.

Child↗