Search PubMed⌕ Search

Biomedical subjects

N Kaneda

Publications and source records attributed to N Kaneda.

At least 109 records · Page 6Linked to original sources
PubMed1986

Simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue by high-performance liquid chromatography with electrochemical detection.

A simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue was developed by using high-performance liquid chromatography with electrochemical detection. These compounds are analysed in a single chromatographic run within 30 min with a simple sample clean-up procedure. The detection system consists of two electrochemical detector cells aligned in series: a glassy-carbon electrode for catecholamines and serotonin, and a platinum electrode for acetylcholine and choline. For the detection of the latter compounds, they were converted enzymatically into hydrogen peroxide through a column reactor with immobilized acetylcholinesterase and choline oxidase. A column of boronic acid gel was placed just ahead of the immobilized enzyme column to remove catecholamines, which caused interfering responses on the platinum electrode. Two equivalent analytical columns and a column switching were employed to speed up the serotonin assay. Simultaneous determination of these major neurotransmitters in rat brain regions was successfully carried out with the system described.

Acetylcholine↗
PubMed1985

Action of cobra venom cardiotoxin on chick embryonal fibroblasts transformed with a temperature-sensitive mutant of Rous sarcoma virus.

The cytolytic action of cardiotoxin analogue III from the venom of the Formosan cobra on chick embryonal fibroblasts transformed with a temperature-sensitive mutant of Rous sarcoma virus was investigated. The 50% effective dose of the toxin for the cells cultured at a non-permissive temperature (41 degrees C) or for noninfected normal cells was about 8 micrograms/ml whereas the value was 2 micrograms/ml for the cells cultured at a permissive temperature (36 degrees C). This indicates that the transformed cells became more susceptible to the cytolytic action of the toxin than the non-transformed cells.

Animals↗
PubMed1985

Highly sensitive assay for acetylcholinesterase activity by high-performance liquid chromatography with electrochemical detection.

A highly sensitive assay for acetylcholinesterase (AChE) activity was devised by high-performance liquid chromatography with electrochemical detection. It is based on the separation of acetylcholine and choline on an octadecylsilane reversed-phase column, followed by their enzymatic conversion into hydrogen peroxide through the post-column reaction with immobilized AChE and choline oxidase. The system is highly sensitive, and the relationship between the peak height and the amount of choline is linear over the range 5 pmol to 5 nmol. When homogenate of bovine caudate nucleus was used as enzyme, the Michaelis constant of the enzyme for acetylcholine was 0.4 mM. The regional distribution of AChE activity in rat brain was examined, and the order of the activity from the highest to the lowest agreed with the reported brain distribution of AChE: striatum, thalamus plus hypothalamus, pons plus medulla oblongata, cerebral cortex, olfactory bulb, and cerebellum.

Acetylcholine↗
PubMed1985

Highly sensitive assay for choline acetyltransferase activity by high-performance liquid chromatography with electrochemical detection.

A highly sensitive assay for choline acetyltransferase activity by high-performance liquid chromatography with electrochemical detection was devised. This assay method is based on the separation of acetylcholine and choline on a Develosil Ph-5 reversed-phase column (a phenyl column), followed by their enzymatic conversion to hydrogen peroxide through post-column reaction with acetylcholinesterase and choline oxidase. The sensitivity of the system is excellent and 5 pmol of acetylcholine enzymatically formed could be detected. The linearity between the peak height and the amount of acetylcholine was observed over the range of 5 pmol to 5 nmol. Some enzymatic properties were investigated by using a soluble fraction of bovine caudate nucleus as enzyme. The Michaelis constants of the enzyme for choline and acetyl coenzyme A were 0.3 mM and 0.03 mM, respectively. The enzyme exhibited the maximum activity over the pH range 7.4-9.5. The regional distribution of choline acetyltransferase activity in rat brain was examined. The order of the activity from the highest to the lowest agreed with the reported brain distribution of the enzyme: striatum, pons plus medulla oblongata, cerebral cortex, thalamus plus hypothalamus, olfactory bulb and cerebellum.

Acetylcholine↗
PubMed1984

Amino acid sequence of cytotoxin IIa isolated from the venom of the Indian cobra (Naja naja).

A cytotoxic basic polypeptide, designated as cytotoxin IIa, was purified to homogeneous state from the venom of the Indian cobra (Naja naja) by a combination of gel filtration on Sephadex G-50, CM-cellulose chromatography, and fast protein liquid chromatography. Cytotoxin IIa is a single polypeptide consisting of 60 amino acid residues with four intramolecular disulfide linkages. The toxin showed high cytotoxicity toward Yoshida sarcoma and ascites hepatoma cells as did cytotoxins I and II isolated from the same venom. Analysis of the amino acid sequence revealed that cytotoxin I, IIa, and II are highly homologous in their primary structures and that cytotoxin IIa differs from cytotoxin I only in having Phe 25 and Val 52 in place of Tyr 25 and Glu 52 residues.

Amino Acid Sequence↗
PubMed1983

Major proteins released by a protein-producing bacterium, Bacillus brevis 47, are derived from cell wall protein.

B. brevis 47 secretes a vast amount of protein consisting mainly of two kinds with approximate molecular weights of 130,000 and 150,000. The two major extracellular proteins were indistinguishable from those of cell wall protein by SDS-polyacrylamide gel electrophoresis. Based on the results of analysis of amino acid composition, limited proteolysis followed by electrophoresis, and the cross-reactivity of antisera, we conclude that the 130K and 150K extracellular proteins are derived from the respective cell wall proteins. Furthermore, the NH2-terminal amino acid analysis suggests that the two major extracellular proteins are released from the cell wall without any modification of the NH2-terminal portion.

Amino Acids↗
PubMed1983

Change in aggregation state of insulin upon conjugation with 5-dimethylaminonaphthalene-1-sulfonyl group.

Change in aggregation state of insulin upon conjugation with 5-dimethylaminonaphthalene-1-sulfonyl (DNS) group was investigated at neutral pH. DNS group was introduced exclusively into B1 phenylalanine, the N-terminus of the B-chain of insulin. The association state of insulin shifted toward a more highly aggregated one upon conjugation, depending on the mole fraction (d) of DNS group to insulin monomer; at d equal 0.3 the equilibrium between dimer and hexamer was dominant over the range of 1-600 microM, while at d equal 1.0-1.5 DNS-insulin formed a larger aggregate (dodecamer) which is stable over the range of 67-600 microM. The dissociation constant of dimer-hexamer equilibrium at d=0.3 was evaluated to be 2.5 x 10(-10) M2 from the fluorescence anisotropy of the DNS group, which was about one order of magnitude smaller than that of the dimer-hexamer equilibrium in native insulin. Spectroscopic data and fluorescence decay analyses indicated that there exist at least two different environments surrounding the dye bound to B1 phenylalanine and that they are both relatively hydrophilic. It is considered that the major part of DNS group has excitation and emission maxima at longer wavelength with relatively low quantum yield, while the minor part has excitation and emission maxima at shorter wavelengths with relatively high quantum yield. The fluorescence lifetime of the dye was modified by the change in quaternary structure of DNS-lifetime of the dye was modified by the change in quaternary structure of DNS-insulin. Remarkable depolarization of DNS fluorescence was observed at d equal 1.0 and d equal 1.5 due to energy transfer between DNS groups conjugated to B1 phenylalanine in the hexamer or the dodecamer. Critical transfer distance for inter-DNS energy transfer was evaluated to be 15 A. From the molecular model of the insulin crystal, this energy transfer is ascribed to the close proximity, within about 15 A, between DNS groups in dimer units of the hexamer or the dodecamer.

Amino Acids↗
PubMed1983

[The present status of mass population screening for cancer of the uterine cervix and the therapeutic results in the detected cases in Miyagi Prefecture].

The mass screening for cervical cancer in the population of Miyagi Prefecture has covered the total 1,078,999 women by March 1981. Invasive cancer of the uterine cervix was discovered in 1032 women, carcinoma in situ in 829. 2) By methods of the examination, the institutional examination was on 72.5% of the examined in 1980. 3) With regards to the rates of examinees by age and the detection rate of the cancer of uterine cervix in 1980, the low detection rate in the age range of 30-50 years whose examination rate was over 20% compares against the high detection rate in the high age range, whose examination rate was low. 4) CAI in years 1965, 1970, 1975 and 1980 were 60.0, 116.7, 193.2 and 229.8 respectively. 5) The 302 cases of invasive cancer detected and treated before the end of 1974 were classified in clinical stages and investigated the results of treatment. The detection in Stage I was in 84.1%, this high percentage being a main factor to make the total treatment results of uterine cervical cancer pushed up to the high figure.

Female↗
PubMed1980

Pulse fluorimetry of N-(1-pyrenesulfonyl)dipalmitoyl-L-alpha-phosphatidylethanolamine in concanavalin A-stimulated human lymphocytes.

Human peripheral lymphocytes were cultured with a fluorescent probe, N-(1-pyrenesulfonyl)dipalmitoyl-L-alpha-phosphatidylethanolamine, and with concanavalin A. Fluorescence microscopic observations revealed that in lymphoblasts, pyrenesulfonyl dye was distributed mainly in vacuoles whereas in normal cells cultured without concanavalin A the dye was distributed exclusively in plasma membranes. The fluorescence spectra of the pyrenesulfonyl group incorporated into the cells exhibited two emission maxima, band A (monomer fluorescence of the pyrenesulfonyl group at about 400 nm) and band B (dimer fluorescence of the dye at about 500 nm). The values of the fluorescence lifetime measured at bands A and B indicated that in the absence of concanavalin A, the environment surrounding the pyrenesulfonyl group at the lipid/water interface became more hydrophilic with cultivation time. Concanavalin A made the environment of the interface more hydrophobic than that of lymphocytes cultured without concanavalin A. Fluorescence polarization measured at band A revealed that the mobility of pyrenesulfonyl monomers at the aqueous interface of the membranes was reduced upon concanavalin A stimulation.

Cell Membrane↗
PubMed1977

Primary structures of cardiotoxin analogues II and IV from the venom of Naja jaja atra.

Cardiotoxin analogues II and IV were isolated from the venom of Naja naja atra by gel filtration on Sephadex G-50 followed by CM-cellulose chromatography. The venom contains at least four cardiotoxin analogues that account for about 54% of the weight of the lyophilized crude venom. These four cardiotoxin analogues, named cardiotoxin analogues I, II, III, and IV, show strong cytotoxicity to Yoshida sarcoma cells but the lethal toxicity is one-order less. These toxins contain 60 amino acid residues in a single peptide chain. Cardiotoxin analogue IV differs from cardiotoxin analogue II only by the presence of arginine in place of a leucine residue at position 1. A comparison of the amino acid sequences of these toxins with that of cobrotoxin, a neurotoxin containing 62 amino acid residues obtained from the same snake venom, shows that about 20 amino acid residues, including 8 half cystine residues, are identical, assuming 3 residues deletion and 2 residues insertion in the cardiotoxin molecule.

Amino Acid Sequence↗