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Biomedical subjects

A Nakamura

Publications and source records attributed to A Nakamura.

At least 19 recordsLinked to original sources
PubMed1992

Differences of draining lymph node cell proliferation among mice, rats and guinea pigs following exposure to metal allergens.

Contact sensitivities of three well known metal allergens (nickel sulfate, potassium dichromate and cobalt chloride) were examined using the local lymph node assay in CBA/N mice, F344 rats and Hartley guinea pigs. The effect of various species sera on lymph node cell (LNC) proliferation was also investigated. Exposure to potassium dichromate and cobalt chloride induced significant LNC proliferative responses in the three species. The LNC responses to potassium dichromate in the rats were higher than those in the mice and guinea pigs. Mice exhibited the highest response to cobalt chloride among the three species, whereas, exposure to nickel sulfate failed to induce a marked LNC proliferation. Increased draining lymph node weights and LNC numbers were also observed following exposure to the metal salts. However, these parameters were less sensitive compared with the LNC proliferative response. There was a large difference in the lymph node weight between individual guinea pigs. The [methyl-3H]thymidine incorporation into LNC of each species cultured in the presence of the homologous serum in vitro was lower than in the presence or absence of fetal calf serum. However, there was no significant difference in stimulation indices among the different culture conditions. The local lymph node assay may be performed in rats as well as in mice for the detection of metal allergens.

Allergens↗
PubMed1992

Gene cloning and characterization of a novel extracellular ribonuclease of Bacillus subtilis.

An extracellular nuclease gene of Bacillus subtilis was cloned in the same organism by detecting the amplified enzyme activity, which was secreted from the transformant cells on an RNA-containing agar medium. An open reading frame encoding 289 amino acids was identified within the cloned fragment. The transcriptional initiation site was determined by nuclease S1 mapping and the promoter region showed similarity to the conserved recognition sequences for the E sigma A and/or E sigma E RNA polymerases. The production of the nuclease by the B. subtilis transformants greatly depends on the liquid medium used. SDS/PAGE analysis of the purified enzyme showed two adjoining bands of molecular mass about 32 kDa, and the NH2-terminal amino acid sequence analysis suggested that the NH2-terminal portion of the nuclease was subjected to a limited proteolysis after or during secretion. The nuclease was uniquely characterized as a Mg(2+)-activated ribonuclease which hydrolyzes RNA apparently nonspecifically into oligonucleotides with 5'-terminal phosphate. The deduced amino acid sequence of this enzyme shows no obvious similarity with other nuclease sequences.

Amino Acid Sequence↗
PubMed1992

Cloning and characterization of gastrin receptor from ECL carcinoid tumor of Mastomys natalensis.

We report here the cDNA cloning of a putative gastrin receptor from enterochromaffin-like (ECL) carcinoid tumor of Mastomys natalensis. For this study, we used the polymerase chain reaction technique to amplify transmembrane domain sequences related to rat pancreatic cholecystokinin (CCK)-A receptor from the ECL tumor cDNA library. The amino acid sequence deduced from the cloned cDNA showed 85.7% and 49.0% identity to canine parietal cell gastrin receptor and rat pancreatic CCK-A receptor, respectively. Ligand binding studies using COS-7 cells transfected with the cDNA showed the same binding specificity for gastrin and CCK-8 as the gastrin receptor on the Mastomys carcinoid tumor membrane. Both gastrin and CCK-8 elevated free cytosolic calcium concentration in COS-7 cells expressing the cloned receptor. RNA blot analysis revealed the expression of the gastrin receptor in both Mastomys stomach and brain.

Amino Acid Sequence↗
PubMed1992

Calcitonin inhibits the growth of human gastric carcinoma cell line KATO III.

Calcitonin has a wide variety of actions on gastrointestinal function. In this study, we investigated the effects of calcitonin on the growth of human gastric carcinoma cell line KATO III in comparison with those of calcitonin gene-related peptide (CGRP). Calcitonin, but not CGRP, significantly and dose-dependently inhibited the growth of KATO III cells. This inhibition of cell growth was accompanied by an increase in cyclic AMP production. The proliferation of KATO III cells was also inhibited by forskolin and dibutyryl cyclic AMP, although agents which do not stimulate cyclic AMP production had no effect. Furthermore, in the presence of GTP, calcitonin stimulated adenylate cyclase activity in KATO III cell membranes, and this increase was reduced in the absence of GTP. On the other had, neither calcitonin nor CGRP enhanced the turnover of inositolphospholipid or the intracellular Ca2+ level. In addition, 125I-labeled human calcitonin was specifically bound to KATO III cell membranes, and this binding was dose-dependently displaced by unlabeled calcitonin but not CGRP. Furthermore, the specific binding of 125I-labeled human calcitonin to KATO III cell membranes was significantly reduced by addition of GTP but not ATP. These results suggest that calcitonin inhibits the growth of human gastric carcinoma cell line KATO III by stimulating cyclic AMP production via a GTP-dependent process coupled to specific calcitonin receptors.

Adenylyl Cyclases↗
PubMed1992

Functional relationships between cyclodextrin glucanotransferase from an alkalophilic Bacillus and alpha-amylases. Site-directed mutagenesis of the conserved two Asp and one Glu residues.

Comparison of the amino acid sequences of cyclodextrin glucanotransferases (CGTases) with those of alpha-amylases revealed that two Asp and one Glu residues, which are considered to be the catalytic residues in alpha-amylases, were also conserved in CGTases. To analyze the function of the three conserved amino acid residues in CGTases, site-directed mutagenesis was carried out. The three mutant CGTases, in which Asp229, Glu257 and Asp328 were individually replaced by Asn or Gln, completely lost both their starch-degrading and beta-cyclodextrin-forming activities, whereas another mutant CGTase, in which Glu264 was replaced by Gln, retained these activities. The three inactive enzymes retained the ability to be bound to starch. These results suggest that Asp229, Glu257 and Asp328 play an important role in the enzymatic reaction catalyzed by CGTase and that a similar catalytic mechanism is present in both CGTases and alpha-amylases.

Aspartic Acid↗
PubMed1992

A synthetic analogue for the active site of plant-type ferredoxin: two different coordination isomers by a four-cys-containing [20]-peptide.

The (Fe2S2)2+ complex of an artificial 20-peptide ligand, Ac-Pro-Tyr-Ser-Cys-Arg-Ala-Gly-Ala-Cys-Ser-Thr-Cys-Ala-Gly-Pro-Leu-Leu-T hr-Cys- Val-NH2, containing an invariant Cys-A-B-C-D-Cys-X-Y-Cys (A, B, C, D, X, Y = amino acid residues) fragment of plant-type ferredoxins was synthesized by a ligand exchange method with [Fe2S2(S-t-Bu)4]2-. 1H-nmr spectroscopic and electrochemical data of the complex indicate the presence of two coordination isomers. One of them having a Cys-X-Y-Cys bridging coordination to the two Fe(III) ions, has the (Fe2S2)2+ core environment similar to those of the denatured plant-type ferredoxins and exhibits a positive shifted redox potential at -0.64 V vs saturated colonel electrode (SCE) in N,N-dimethylformamide (DMF). Another isomer with the Cys-A-B-C-D-Cys bridging coordination shows a negative redox potential at -0.96 V vs SCE in DMF.

Amino Acid Sequence↗
PubMed1992

Comparative studies by cell culture and in vivo implantation test on the toxicity of natural rubber latex materials.

Colony assay using V79 cells, the agar diffusion assay with L929 cells, and the 7-day rabbit muscle implantation test were employed to evaluate the cytotoxicity and tissue toxicity of natural rubber latex (NRL) materials. The in vivo implantation test showed that, among 13 histological parameters, thickness of inflammatory layer was the most useful index to evaluate tissue responses quantitatively. A comparison of the in vivo and in vitro parameters revealed the following correlations between the thickness of the inflammatory layer and cytotoxicity indices: Colony assay of the extracts, IC50: r = 0.80; Agar diffusion assay, Zone index: r = 0.73; Lysis index: r = 0.61. From these results, it appears that the colony assay provides a more reliable prediction of the tissue response than the agar diffusion assay.

Animals↗
PubMed1992

Difference in tumor incidence and other tissue responses to polyetherurethanes and polydimethylsiloxane in long-term subcutaneous implantation into rats.

The long-term (1- and 2-year) adverse tissue responses including tumor formation by subcutaneous implantation of polyurethanes (PUs) and silicone (Sil) films into rats were compared. The weight-averaged molecular weights (Mw) of the PUs prepared from 4,4'-diphenylmethanediisocyanate, poly(tetramethyleneglycol) of Mn = 1000 and 1,4-butanediol are 220,000 (U-4), 124,000 (U-6), and 55,600 (U-8). The 50:50 mixed film of U-6 and silicone (U-6/sil) was prepared by roll-mixing of the noncured silicone and the U-6 solution followed by evaporation of the solvent and heat-curing at 70 degrees C. The tissue responses around implants were classified into four groups as follows: (A) tumor, (B) atypical cell proliferation accompanied by preneoplastic changes, (C) cell proliferation without preneoplastic changes, (D) no obvious responses. In both implantation periods, the PUs gave higher incidents of the adverse responses including tumor formation in comparison to Sil. No significant molecular weight-dependent trend was found in a 1-year study using U-4, 6, and 8. Significant PU-dose-dependent trends were found in a 2-year study: the total active incidence (A+B+C), U-6(22/29) greater than U-6/sil(11/29) greater than sil(7/28); tumor incidence (A), U-6(11/29) greater than U-6/sil(2/29) = sil(2/28). No detectable amounts of 4,4'-methylenedianiline (MDA) were found in the PUs. The methanol extracts from the PUs were negative in the mutagenicity tests. These indicate no relationship between the tumor formation by the PU films and the mutagenicities of the chemicals (mainly oligomers) leached from the PUs.

Aniline Compounds↗
PubMed1992

Effects of injecting fibronectin and antifibronectin antibodies on cushion mesenchyme formation in the chick. An in vivo study.

During heart development in the chick some of the endocardial cells that cover the cushion areas leave the cushion endocardium, seed the underlying cardiac jelly, and are transformed into mesenchyme. Cushion mesenchymal (CM) cells migrate from the endocardium toward the myocardium using the cardiac jelly as substratum. Developing cushions have been microinjected with fibronectin (FN), antifibronectin antibodies (AbFN), and four synthetic peptide probes. Two of these peptides (P7 and P10) contained the sequence Arg-Gly-Asp-Ser (RGDS), while the other two (P15 and PColl) did not. Cushion area, individual cell area, cell density, cell orientation and a factor of form were evaluated in both experimental and control cushions. CM cell migration was inhibited by FN and AbFN, only partially inhibited by P10 and unaffected by P7. Cushions injected with P15 and PColl were unaffected. These results can be explained by steric modifications of the extracellular matrix, that may render cardiac jelly nonpermissive for CM cell migration, or by interaction of the substances injected at the endocardial cell surface. Migrating CM cells do not present any preferential orientation in any particular direction. CM cell migration seems to depend upon intrinsic migratory behaviour and the presence of FN at the CM cell surface. The enforcement of the direction of CM cell migration does not appear to rely upon matrix signals but be the result of randomly migrating cells becoming distributed more evenly in the matrix.

Amino Acid Sequence↗
PubMed1992

Factors contributing to deficiencies in cell-mediated immunity in esophageal cancer patients.

Based on the previous data which indicated a preoperative decrease in cell-mediated immunity (CMI) is associated with the occurrence of infectious complications following surgery on patients with esophageal cancer, we examined possible factors contributing to a decrease in CMI levels. A multiple linear regression analysis was made on data from 76 patients with esophageal cancer and 53 with gastric cancer as the control. In patients with esophageal cancer, both protein-calorie malnutrition (PCM) and age factor contributed to a decrease in CMI, although the contribution of the latter was weak while the stage of the cancer and the grade of dysphagia showed no such contribution. The PCM and stage of the cancer were contributing factors in patients with gastric cancer. Thus, these results indicate that PCM and old age, and not the presence of malignant tumors, play a significant role in deficiency in CMI in patients with esophageal cancer.

Age Factors↗
PubMed1992

Primary structure of chain I of the heterodimeric hemoglobin from the blood clam Barbatia virescens.

The blood clam Barbatia virescens has a heterodimeric hemoglobin in erythrocytes. Interestingly, the congeneric clams B. reeveana and B. lima contain quite different hemoglobins: tetramer and polymeric hemoglobin consisting of unusual didomain chain. The complete amino acid sequence of chain I of B. virescens has been determined. The sequence was mainly determined from CNBr peptides and their subpeptides, and the alignment of the peptides was confirmed by sequencing of PCR-amplified cDNA for B. virescens chain I. The cDNA-derived amino acid sequence matched completely with the sequence proposed from protein sequencing. B. virescens chain I is composed of 156 amino acid residues, and the molecular mass was calculated to be 18,387 D, including a heme group. The sequence of B. virescens chain I showed 35-42% sequence identity with those of the related clam Anadara trapezia and the congeneric clam B. reeveana. An evolutionary tree for Anadara and Barbatia chains clearly indicates that all of the chains are evolved from one ancestral globin gene, and that the divergence of chains has occurred in each clam after the speciation. The evolutionary rate for clam hemoglobins was estimated to be about four times faster than that of vertebrate hemoglobin. We suggest that blood clam hemoglobin is a physiologically less important molecule when compared with vertebrate hemoglobins, and so it evolved rapidly and resulted in a remarkable diversity in quaternary and subunit structure within a relatively short period.

Amino Acid Sequence↗
PubMed1992

Effects of islet amyloid polypeptide (amylin) and calcitonin gene-related peptide (CGRP) on glucose metabolism in the rat.

In this study, we compared the effects of islet amyloid polypeptide (IAPP) and calcitonin gene-related peptide (CGRP) on glucose metabolism both in vivo and in vitro in the rat. Intravenous injection of rat CGRP caused a significant increase in plasma glucose concentration with a simultaneous increase in plasma insulin levels, whereas neither IAPP-NH2 nor IAPP-COOH had any effect. Moreover, intravenous infusion of CGRP decreased tolerance to intragastric administration of glucose (O-GTT) without altering plasma insulin levels, but again IAPPs had no effect. On the other hand, 125I-[Tyr0]rat CGRP specifically bound to the liver plasma membrane, and not only CGRP but also IAPP-NH2 dose-dependently displaced the specific binding of 125I-[Tyr0] CGRP, whereas IAPP-COOH had no effect. Conversely, CGRP as well as IAPP-NH2 but not IAPP-COOH evoked dose-dependent activation of adenylate cyclase in the membranes, and these effects were significantly inhibited by a CGRP receptor antagonist, human CGRP-I(8-37). However, neither CGRP nor IAPP-NH2 had any effect on glucose production in rat isolated hepatocytes. These results suggest that (1) IAPP-NH2 but not IAPP-COOH induces adenylate cyclase activation via CGRP receptors on rat liver plasma membranes, and (2) CGRP might not involve its action on the liver in the changes of glucose metabolism.

Amyloid↗
PubMed1992

Positive feedback action of pituitary beta-endorphin on acupuncture analgesia afferent pathway.

Potentials in the final sector of the afferent pathway from the acupuncture point (AP) were enhanced by intraperitoneal 0.5 mg/kg morphine without changing the threshold of AP stimulation and greatly decreased by hypophysectomy. The decreased potentials were restored to the control level by morphine (0.5 mg/kg, IP). Potentials evoked in the final sector of the afferent pathway from the nonacupuncture point (NAP) by NAP stimulation after lesion of the analgesia inhibitory system were greatly enhanced by corticotropin (ACTH) (0.25 mg/kg, IP) and greatly decreased by hypophysectomy. Diminished potentials were restored to the control level by ACTH (0.25 mg/kg, IP). Both morphine (0.5 mg/kg, IP) and ACTH (0.25 mg/kg, IP) produced analgesia, but morphine did not affect acupuncture analgesia (AA) and ACTH did not affect nonacupuncture point stimulation-produced analgesia (NAA). All analgesia, that due to 0.5 mg/kg morphine or 0.25 mg/kg ACTH, AA, and NAA were abolished by hypophysectomy. The abolished AA and NAA were restored by 0.5 mg/kg morphine and 0.25 mg/kg ACTH, respectively. Hence, beta-E and ACTH liberated from the pituitary gland by stimulation of an AP and NAP may act as positive feedback on the AA and NAA afferent pathways, respectively.

Acupuncture Analgesia↗
PubMed1992

Serotonergic neurons in the brainstem modulate animal hypnosis.

We previously found that the center of animal hypnosis production in the rabbit is located around the locus ceruleus and brachium conjunctivum (LC-BC) of the brainstem. The involvement of serotonergic neurons in this area of animal hypnosis was investigated by microinjection of serotonin into these regions. The duration of animal hypnosis (DAH) induced by inversion was diminished to about 65% of the controls by serotonin microinjection into the LC-BC and microinjection of methysergide prolonged the DAH to 3.2 times that of the controls. Flexor muscle contraction (CFM) of the upper extremities induced by electrical stimulation of the motor cortex was enhanced by serotonin. In normal rabbits, hard pressure on the ear base or the lumbar paravertebral area reduced CFM and this effect was partially antagonized by serotonin microinjected into the LC-BC. The results suggest that serotonergic neurons in the LC-BC modulate animal hypnosis.

Animals↗
PubMed1992

Detection of contact sensitivity of metal salts using the murine local lymph node assay.

The local lymph node assay (LLNA) is a predictive test for the detection of contact allergens. Nickel and chromium sensitization are common cases in man. However, in a previous study topical application of nickel sulfate and potassium dichromate in aqueous solution failed to induce activation in the draining lymph node. This study describes the application of LLNA to evaluate the contact sensitivity of metal salts. The metal salts were applied in dimethylsulfoxide or aqueous ethanol solution. In some experiments, the skin of the ears was gently abraded using a needle prior to application of metal salts. The ability of seven metal salts to induce lymph node cell (LNC) proliferation was compared. Nickel, cobalt, chromium and copper salts increased LNC proliferation, whereas zinc, manganese and iron salts failed to induce LNC proliferation in this assay.

Animals↗
PubMed1992

alpha-Hydroperoxy diethyl peroxide, an unusual natural compound isolated from an ascidian (Phallusia mammillata): acute toxicity in DDY mice.

alpha-Hydroperoxy diethyl peroxide, a novel compound found in the tunic of ascidians, has two peroxide moieties per molecule. Since ascidians are a widely served food item in Japan, human exposure to this compound potentially exists in the seafood preparation industries. No toxicological data have so far been published on this compound, and so we determined the intraperitoneal 6-day LD50 in mice and conducted histopathological examinations. The 6-day LD50 was found to be 199 mg/kg with 95% confidence limits of 126-314 mg/kg. Histopathological examination revealed necrosis induced in a variety of cells that had been directly exposed to the compound. These cells included hepatocytes, parenchymal pancreatic cells and fat cells. It is concluded that direct contact with this compound is likely to elicit cellular necrosis of various organs. The specific toxicological effects are probably dependent on the route of exposure.

Animals↗
PubMed1992

A method for identifying causative chemicals of allergic contact dermatitis using a combination of chemical analysis and patch testing in patients and animal groups: application to a case of rubber boot dermatitis.

A 63-year-old woman developed allergic contact dermatitis from rubber boots. Initial investigation, by patch testing in the patient and chemical analysis of the causative rubber boots, revealed that mercaptobenzothiazole (MBT) and dibenzothiazyl disulfide (MBTS) were the causative chemicals. Subsequent investigations were performed by patch testing in animal groups. An extract of the causative rubber boots, MBT and MBTS were used for sensitization of guinea pigs by the guinea pig maximization test (GPMT). 3 animal groups, A (with the boot extract), B (with MBT) and C (with MBTS) were successfully prepared. The boot extract was fractionated by column chromatography and thin-layer chromatography (TLC). Each fraction was subjected to patch testing in the animal groups. Positive reactions in all groups would show that the active fractions contained MBT-type compounds, whereas a positive reaction in group A but negative ones in group B and C would show that the active fractions did not contain any MBT-type compounds. Each fraction was then analyzed by gas chromatography (GC), GC-mass spectrometry (GC-MS), direct inlet-MS (DI-MS) and high-performance liquid chromatography (HPLC). By this investigation, we found not only known allergens (MBT, MBTS), but also unknown allergens: S-substituted MBT-type compounds and styrenated phenol (SP). Thus, SP was shown to be a candidate as a human sensitizer even though the patient did not react to it.

Allergens↗