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Biomedical subjects

K Miyazaki

Publications and source records attributed to K Miyazaki.

At least 595 records · Page 33Linked to original sources
PubMed1988

2,4-Dinitrophenylhydrazides of polysialogangliosides.

Treatment with 2,4-dinitrophenylhydrazine HCl in the presence of dicyclohexylcarbodiimide, converts gangliosides to their dinitrophenylhydrazides. This derivatization is the basis of a useful method for HPLC determination of gangliosides (K. Miyazaki, N. Okamura, Y. Kishimoto and Y. C. Lee (1986) Biochem. J. 235, 755-761). This procedure, however, yields two different GT1b products. By characterizing these two products using plasma desorption mass spectrometry, proton magnetic resonance and other chemical and physical techniques, we found that either one or two of the three sialic acid carboxyl groups in GT1b, were converted to dinitrophenylhydrazides. The remaining underivatized carboxyl groups formed lactones with hydroxyl groups from other carbohydrate residues. Also, while sialic acid residues of GD1a were fully derivatized, only one sialic acid in GD1b, two sialic acids in GT1a and two in GQ1b were converted to dinitrophenylhydrazides, the remaining carboxyl groups probably forming lactones. Sialic acid residues between galactose of the gangliotetraose chain and another sialic acid in polysialogangliosides appear to be underivatized possibly because of steric hindrance.

Chemical Phenomena↗
PubMed1988

Pancreatic polypeptide secretion from the isolated perfused ventral and dorsal areas of the rat pancreas.

The secretion of pancreatic polypeptide (PP) by neuropeptides [vasoactive intestinal polypeptide, gastrin-releasing peptide (GRP), neuromedin B, and neuromedin C] and carbachol was investigated using the isolated perfused ventral part of the rat pancreas, with or without atropine, and a specific radioimmunoassay for rat PP. The release of PP from the dorsal part of the rat pancreas by GRP and carbachol was also studied. Carbachol and GRP stimulated, in a dose-dependent manner ranging from 10(-6) to 10(-9) M, PP secretion from the ventral part of the pancreas. Maximum responses were observed with 10(-7) to 10(-8) M carbachol and 10(-7) M GRP. The response with 10(-7) M carbachol was biphasic and was abolished by 10(-5) M atropine. The response with 10(-7) M GRP, however, was transient, monophasic, and not inhibited by atropine. Vasoactive intestinal polypeptide (10(-7) M) stimulated a weak PP secretion that was not inhibited by atropine. The potency of GRP and vasoactive intestinal polypeptide relative to carbachol at 10(-7) M was 24% and 7%, respectively. Neuromedin C had almost the same bioactivity as GRP with respect to the release of PP, whereas neuromedin B was one-third less potent than GRP and neuromedin C. The relative secretion of PP from the dorsal compared with the ventral part of the pancreas after stimulation by 10(-7) M carbachol and GRP was 19% and 22%, respectively. These data show that peptidergic nerves may play some role in PP secretion. In addition, the main source of PP to the systemic circulation may be the ventral part of the pancreas.

Animals↗
PubMed1988

Gastrin-releasing peptide stimulates cholecystokinin secretion in perfused rat duodenum.

We examined the effect of porcine gastrin-releasing peptide (GRP-27) and other analogous neuropeptides on cholecystokinin (CCK) secretion from the isolated perfused rat duodenum. GRP-27 stimulated CCK secretion in a monophasic pattern and in a dose-dependent manner ranging from 10(-9) M to 10(-6) M, and 10(-7) M of GRP-27 led to an increment of 442 +/- 120.8 fmol/3 min. The stimulatory effect of GRP-27 on CCK was not inhibited by 10(-5) M of atropine. 10(-7) M of neuromedin C and B, analogs of GRP, stimulated CCK secretion to increments of 382 +/- 64.1 and 289 +/- 47.2 fmol/3 min, respectively. Carbachol (10(-9) to 10(-6) M), VIP (10(-9)M), secretin (10(-9)M) and glucose (11 mM) did not stimulate CCK secretion, and the addition of atropine (10(-5)M) to them led to no significant changes. These results suggest that GRP may directly stimulate CCK secretion from the duodenum and work as a non-cholinergic, peptidergic neurotransmitter.

Animals↗
PubMed1988

Distribution of pancreatic polypeptide-like immunoreactivity in rat tissues.

The distribution of pancreatic polypeptide (PP)-like immunoreactivity (LI) in rat tissue was determined by a specific radioimmunoassay (RIA) after extraction with boiling 1 N acetic acid. The concentration of PP-LI in the ventral area of the pancreas (0.917 +/- 0.106 micrograms/g tissue) was about 10 times greater than that in the dorsal area of the pancreas (0.085 +/- 0.006 micrograms/g tissue). Extrapancreatic PP-LI was present in the colon (0.034 +/- 0.010 micrograms/g tissue) and rectum (0.019 +/- 0.001 micrograms/g tissue). The remainder of the gastrointestinal tract, the lung, kidney, liver, spleen, heart, adrenal gland, and central nervous system contained no measurable PP-LI. Reverse-phase high performance liquid chromatography analysis of the PP-LI materials from the pancreas, colon, and rectum revealed one peak which corresponds to the rat PP standard, under conditions of elution which clearly separated PP, NPY, PYY. These results show that distribution of PP-LI in the rat is different from other known distributions in the PP family of peptides.

Animals↗
PubMed1988

DNA repair synthesis in primary culture of bovine bile duct epithelial cells induced by chemical agents in relation to bile duct cancer.

Unscheduled DNA synthesis (UDS) was measured autoradiographically in a primary culture of extrahepatic bile duct epithelial cells of Holstein cows following exposure to chemicals known to be capable of developing bile duct cancers in experimental animals, i.e., N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, CAS No. 70-25-7), N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG, CAS No. 4255-77-6), 20-methylcholanthrene (MCA, CAS No. 56-49-5), N-nitrosodimethylamine (DMN, CAS No. 62-75-9) and aflatoxin B1 (AFB1, CAS No. 1162-65-8). MNNG and ENNG induced UDS without addition of S9 mixture. MCA elicited UDS only if S9 mixture was added. Regardless of the presence or absence of S9 mixture, DMN failed to induce UDS. DNA repair by AFB1 was enhanced by the presence of S9 mixture. Therefore, for MNNG, ENNG and high doses of AFB1 activation by the liver is not necessary to exert genotoxic effects and they seem to be capable of direct action on bile duct epithelial cells in the presence of a bilioenteric fistula or anastomosis.

Animals↗
PubMed1988

Multiple forms of growth inhibitors secreted from cultured rat liver cells: purification and characterization.

It was found that a non-tumorigenic epithelial cell line from the liver of a Buffalo-strain rat (BRL) secreted into the culture medium various inhibitors of the growth of BRL and RSV-BRL (tumorigenic BRL transformed by infection of Rous sarcoma virus). The secreted inhibitors were classified into two types: one inhibited the growth of BRL to a greater extent than that of RSV-BRL (non-tumorigenic BRL growth inhibitor, NGI), and the other, vice versa (tumorigenic BRL growth inhibitor, TGI). Two NGI (NGI-I and NGI-II) and two TGI (TGI-I and TGI-II) were highly purified from the serum-free conditioned medium. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis without 2-mercaptoethanol, NGI-I and II gave protein bands with molecular weights (Mr) of 56,000 and 21,000, respectively. TGI-I and II gave a band that migrated faster than bromophenol blue marker dye, but they did not pass through an ultrafiltration membrane with an Mr cutoff of 5,000. In the presence of a reducing reagent, only NGI-II showed a decrease of Mr, from 21,000 to 11,000. NGI and TGI showed 50% growth inhibition with BRL and RSV-BRL, respectively, at 5-15 ng/ml in the medium containing 10% fetal calf serum. NGI and TGI all were stable to 1 M acetic acid (pH 2.3) and 6 M urea, but labile to 5 mM dithiothreitol or trypsin. Of the eight cell lines tested, NGI-I was most effective on BRL, NGI-II on BRL and HSC-3 (human tongue squamous carcinoma), and both TGI-I and II on RSV-BRL.

Animals↗
PubMed1988

Binding of organic cations to brush border membrane from rat small intestine.

The binding of six organic cations (chlorpromazine, promethazine, imipramine, diphenhydramine, methochlorpromazine and propantheline) to the brush border membrane isolated from rat small intestine has been investigated. The cations were bound to the membrane to varying extents, the order of binding being chlorpromazine greater than promethazine greater than methochlorpromazine greater than imipramine greater than propantheline greater than diphenhydramine. There was no relation between binding and the chloroform-water partition coefficient. Chlorpromazine binding was significantly decreased in the presence of imipramine, methochlorpromazine and propantheline. Anionic compounds (indomethacin and xanthene-9-carboxylic acid) did not affect chlorpromazine binding. High and low affinity binding of the cations to the intestinal brush border membrane was demonstrated with Scatchard plots and Hill plots. Imipramine and methochlorpromazine inhibited chlorpromazine binding at both binding sites. From the results, it was suggested that the organic cations tested were specifically bound to common binding sites on the brush border membrane.

Animals↗
PubMed1988

Transport characteristics of propantheline across rat intestinal brush border membrane.

The transport mechanism of propantheline, an anti-acetylcholine quaternary ammonium compound, has been studied using brush border membrane vesicles isolated from rat small intestine. The uptake of propantheline was facilitated by the transmembrane electrical potential difference (cell interior negative) induced by NaSCN, NaI or valinomycin. But this effect was a secondary action; in the initial phase of propantheline uptake (less than 5 min), there was no facilitating effect. When the transmembrane potential difference was induced after propantheline uptake had reached a steady state, there was an overshoot of the drug. Therefore, it is suggested that the transport of propantheline across the brush border membrane consists of at least two processes. In the first, propantheline rapidly binds to the brush border membrane, in the second it enters into epithelium driven by the negative transmembrane electrical potential difference. Cationic tertiary amines such as chlorpromazine, imipramine and promethazine markedly inhibited propantheline uptake. These results suggest that there is a common absorption process for tertiary amines and quaternary ammonium compounds.

Animals↗
PubMed1988

Effect of chlorpromazine on the permeability of beta-lactam antibiotics across rat intestinal brush border membrane vesicles.

The effect of chlorpromazine on the membrane permeability of beta-lactam antibiotics (benzylpenicillin, ampicillin, cephradine and cephalexin) and actively transported substances (glycylglycine and D-glucose) has been studied using rat intestinal brush border membrane vesicles. Except for cephalexin, the initial uptakes at 25 degrees C of these antibiotics were significantly enhanced in the presence of chlorpromazine. In contrast, the transport of glycylglycine and D-glucose was significantly inhibited. These results suggest that the two groups, drugs and actively transported substances, have a different permeation process. The effect of chlorpromazine concentration on membrane lipid fluidity, as assessed by the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-anilino-8-naphthalene sulphonate (ANS), was also examined. The fluorescence polarization of ANS decreased with increasing concentration of chlorpromazine, while that of DPH increased suggesting an increase of membrane surface fluidity might affect the permeation of beta-lactam antibiotics and actively transported substances in a different manner.

Animals↗
PubMed1988

Interstitial deletion 2q32.1----q34 in a child with half normal activity of ribulose 5-phosphate 3-epimerase (RPE).

High resolution banding analysis showed a de novo interstitial deletion, 46,XX, del(2) (q32.1q34), in a malformed and severely mentally retarded girl aged nine years. Biochemical studies showed that the proband had half normal activities of both erythrocyte isocitrate dehydrogenase (IDH1) and ribulose 5-phosphate 3-epimerase (RPE). It is suggested that the gene for RPE is located on the segment 2q32.1----q34.

Carbohydrate Epimerases↗
PubMed1988

[Experimental knee pain model in rats and analgesic effect of sodium hyaluronate (SPH)].

Using the model of knee pain reaction induced by intra-articular injection of endogenous pain substances, especially bradykinin (BK) in rats, the mechanism of the analgesic effect of sodium hyaluronate (SPH) was investigated. The simultaneous administration of prostaglandin E2 with BK or hyaluronidase digestion of endogenous hyaluronic acid (HA) in our experiments brought remarkable hyperalgesia on BK-induced knee pain. These results suggest that higher sensitivity to the pain reaction is induced in a diseased joint (higher prostaglandin content, lower concentration and molecular size of HA in synovial fluid) than in a normal one. SPH definitely decreased BK-induced pain, and its analgesic effect was observed for a longer period, depending on its dose in pre-treatment and the degree of its distribution in synovial tissues. As the analgesic effect of SPH was observed in the hyaluronidase-treated joint as well, it is suggested that the increasing viscosity of synovial fluid caused by increasing HA concentration can decrease the pain even without normalizing molecular size of HA in the joint. HA oligomer and other compounds with similar viscosity or with similar polyanionic character as SPH showed no analgesic effect. From these results, it seems that the characteristic steric configurations of higher molecular HA are needed for the manifestation of the analgesic effect. SPH seems to show its analgesic effect by covering pain receptors in synovial tissues and holding endogenous pain substances in its molecule.

Analgesics↗
PubMed1988

Presence of high concentration of 7B2 in pleural effusion.

7B2 (a novel pituitary protein) is a secretory protein in the neuroendocrine tissues and an increase in the plasma 7B2 concentration was noted in some patients with various endocrine tumors, including small cell carcinoma of the lung and acromegaly, suggesting that 7B2 is a possible marker for these tumors. Using a radioimmunoassay, the 7B2 concentration was measured in pleural fluid samples obtained from 36 patients with lung cancer and benign pulmonary disease to assess its concentration as a marker for small cell carcinoma of the lung (SCCL) or malignant effusion. 7B2-immunoreactivity (IR) was present in pleural fluid and its concentration was much higher than in plasma. However, there was no significant difference between pleural fluid 7B2 in patients with SCCL and in other histological types of lung carcinoma or in malignant and nonmalignant patients. In the chromatographic analysis of pleural fluid on gel permeation chromatography and reverse-phase high-performance liquid chromatography, there was no molecular heterogeneity between malignant and nonmalignant effusion. These results suggest that pleural fluid 7B2-IR is not a useful marker for SCCL or malignant effusion.

Chromatography, Liquid↗
PubMed1988

Molecular analysis of Lesch-Nyhan syndrome found in Japan.

Molecular analysis of four unrelated patients with Lesch-Nyhan (L-N) syndrome was performed. All four cases had typical clinical features of L-N syndrome, and the activities of hypoxanthine-guanine phosphoribosyltransferase (HPRT) were absent. No structural gene abnormalities were found by Southern blot analysis in all cases, and a decrease of HPRT mRNA was not detected by dot blot analysis in two of the four cases. However, HPRT enzyme proteins were detected by Western blot analysis in all cases. Our results showed that the production of immunologically reactive but enzymatically inactive mutant HPRT protein was pathogenic for all four patients.

Adult↗
PubMed1988

Plasma basal concentration of pancreatic polypeptide (PP) does not increase with ageing in the female Fischer (F-344) conscious rat.

Plasma concentrations of pancreatic polypeptide (PP) were compared for young (6 to 12 month-old) and old (24 to 25 month-old) conscious female Fischer rats by means of a radioimmunoassay. Basal plasma PP concentration was not different between young and old rats, nor was PP release stimulated by cerulein in either age group. The mechanism of PP release in rats is quite different from that in other species such as dogs and humans.

Aging↗
PubMed1988

Synthesis of histargin and related compounds and their inhibition of enzymes.

A novel method for the synthesis of histargin and its analogs is described. It includes two kinds of N-alkylation reactions that prevent the formation of side products. The inhibition of enzymes by these compounds was also measured. Some of the compounds strongly inhibited carboxypeptidase B, carboxypeptidase A, carboxypeptidase N (kininase I), and angiotensin converting enzyme.

Angiotensin-Converting Enzyme Inhibitors↗