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Biomedical subjects

H Nojima

Publications and source records attributed to H Nojima.

At least 163 records · Page 9Linked to original sources
PubMed1986

Primary structure of the alpha-subunit of human Na,K-ATPase deduced from cDNA sequence.

Clones carrying cDNA sequences for the alpha-subunit of the Na,K-ATPase from HeLa cells have been isolated. Nucleotide sequence analysis of the cloned cDNA has revealed the primary structure of this polypeptide, which consists of 1,023 amino acids. The alpha-subunit of the human Na,K-ATPase exhibited 87% homology with its Torpedo counterpart and 98% homology with its sheep counterpart. The six putative transmembrane segments M1-M6 showed higher conservation than the total segments. Total genomic Southern hybridization indicated the existence of at most two copies, possibly only one, of the gene encoding the Na,K-ATPase alpha-subunit in the human genome.

Amino Acid Sequence↗
PubMed1986

Hypersensitivity of acetylcholine receptor in diabetic skeletal muscle to neuromuscular blockers: the effect of myotubes cultured with spinal cord or its extract.

The hypersensitivity of the neuromuscular junctions of diabetic mice to succinylcholine (SuCh), but not to d-tubocurarine (d-TC), was investigated using a cross culture preparation of diabetic skeletal muscle or spinal cord extract with normal tissues. Whether the hypersensitivity is due to the muscle cells themselves was examined using adult muscle of diabetic KK-CAy, prediabetic KK-CAy and normal ddY mice cocultured with embryonic spinal cord of normal ddY mice. The cultured neuromuscular junctions between diabetic KK-CAy muscle and normal ddY spinal cord was hypersensitive to SuCh, but not to d-TC. In contrast, such junctions between prediabetic KK-CAy muscle and normal ddY spinal cord were not hypersensitive to either drug. The involvement of neuronal factors in hypersensitivity to SuCh in diabetic KK-CAy neuromuscular junctions was examined using adult spinal cord extract (SCE) from diabetic KK-CAy and from normal ddY mice. We followed the time course of change in sensitivity of the acetylcholine (ACh) receptors in normal ddY embryonic myotubes to SuCh and d-TC. Both diabetic SCE and normal SCE reduced the sensitivity of myotubes to ACh; the reduction of ACh potential amplitudes by the former was less than that by the latter. Myotubes cultured with diabetic SCE was hypersensitive to both 1.51 microM SuCh and 0.134 microM d-TC. These results suggest that the hypersensitivity of the neuromuscular junctions in diabetic KK-CAy mice to SuCh but not to d-TC is mainly attributable to the diabetic muscle cells themselves.

Animals↗
PubMed1986

Diabetes mellitus-induced hypersensitivity of mouse skeletal muscles to acetylcholine and succinylcholine.

The myopathy in skeletal muscles of genetically diabetic male KK-CAy mice or alloxan-induced diabetic mice was investigated. In these diabetic mice, nerve-stimulated twitch tensions of in situ sciatic nerve-gastrocnemius muscle preparations were inhibited by intraarterially administered succinylcholine (SuCh) to a greater extent than in normal (non-diabetic) ones. Despite the high blood glucose level, at one week after alloxan administration, no hypersensitivity to SuCh was induced in mice, but mice at 2 weeks and 4 weeks after alloxan showed greater sensitivities. In isolated diaphragm muscles of diabetic KK-CAy mice, the acetylcholine (ACh, iontophoretically applied) potential amplitude was greater than in KK-CAy prediabetic muscles. SuCh in diabetic KK-CAy muscles inhibited ACh potentials to a greater extent than in normal ddY muscles. Hill coefficients obtained from the inhibition curve by SuCh of the nerve-stimulation response were decreased by the diabetic state. The sensitivities to d-tubocurarine and alpha-bungarotoxin were the same in both kinds of muscles. Both extrajunctional ACh receptors in denervated muscles of normal ddY and diabetic KK-CAy mice revealed the lower sensitivity to SuCh than junctional receptors in non-denervated normal muscles. In conclusion, diabetic muscles showed the hypersensitivity restricted to SuCh. These phenomena are neither due to glycosylation nor to denervation supersensitivity.

Acetylcholine↗
PubMed1985

Depolarizing neuromuscular blocking action induced by electropharmacological coupling in the combined effect of paeoniflorin and glycyrrhizin.

Twitch tensions of indirectly stimulated diaphragm muscles of mice were blocked by a combination of paeoniflorin (PF) and glycyrrhizin (GLR). The mechanism of this effect was studied electropharmacologically. When twitch responses were completely prevented, miniature end-plate, end-plate, and muscle action potentials were still observed when PF and GLR were combined, suggesting that the mechanism is postsynaptic. Potential amplitudes induced by acetylcholine (ACh), which was injected iontophoretically, were inhibited by about 70% by PF (25 micrograms/ml) plus GLR (75 micrograms/ml), although neither agent alone caused an inhibition. The combined ratio (PF:GLR = 1:3) by concentrations (g/ml) potentiated both the inhibition of ACh potential amplitudes and the depolarization of resting membrane potentials. These results indicate that the effect of combined PF and GLR is to depolarize the muscle membrane and to block ACh-receptor-linked processes. In chemically skinned (saponin-treated) muscles, the tension induced by 0.39 microM of free calcium was inhibited by PF (300 micrograms/ml), but it tended to be increased by GLR (300 micrograms/ml). Caffeine-induced contractures in the skinned muscles was not influenced by PF, and they tended to be decreased with GLR treatment. Thus, in muscles with sustained depolarization, these combined compounds seem to block intracellular Ca2+ movement.

Acetylcholine↗
PubMed1984

High guanine plus cytosine content in the third letter of codons of an extreme thermophile. DNA sequence of the isopropylmalate dehydrogenase of Thermus thermophilus.

In studies on the cause of the extreme stability of the macromolecules of Thermus thermophilus HB8, the leuB gene coding for 3-isopropylmalate dehydrogenase of the leucine synthesis pathway and its flanking regions were cloned and sequenced. The leuB gene of T. thermophilus was expressed in a leuB-less mutant of Escherichia coli, and thermostable dehydrogenase was purified from an extract of the cells. The primary structure of the thermophilic isopropylmalate dehydrogenase was deduced from the nucleotide sequence leuB gene (1017 base pairs) and the amino acid sequence of the peptides isolated from the purified dehydrogenase. The thermophilic dehydrogenase has Mr = 35,968, and the value was close to that determined for the monomer of dehydrogenase (36,000) by gel electrophoresis. The molecular weight of active dimeric dehydrogenase was found to be 73,000 by high speed liquid chromatography. The primary structure of dehydrogenase was consistent with the amino acid composition of the dehydrogenase. In contrast to the isopropylmalate dehydrogenase of E. coli which contains 8 cysteine residues, there was no cysteine in thermophilic isopropylmalate dehydrogenase. The 5'-noncoding region contained a typical Shine-Dalgarno sequence. The guanine plus cytosine content of the coding region was 70.1%, and that of the third letter of the codons was extremely high (89.4%).

3-Isopropylmalate Dehydrogenase↗
PubMed1984

Blocking effects of a new component, paeoniflorigenone, in paeony root on neuromuscular junctions of frogs and mice.

A new monoterpene, paeoniflorigenone (PFG) (100-900 micrograms/ml), which was isolated from paeony roots and identified chemically, suppressed both indirectly and directly stimulated muscle twitchings of frog sciatic nerve-sartorius muscle preparation, and it indirectly stimulated muscle twitchings of phrenic nerve-diaphragm muscle preparations. The suppression effect by PFG (300 micrograms/ml) on twitching was not reversed by neostigmine (60 micrograms/ml) and was restored by washing out of PFG. PFG (150 micrograms/ml) depolarized the diaphragm muscle membranes by 10 mV and did not change the electrotonic potentials. PFG (100 micrograms/ml) inhibited weakly acetylcholine (5 micrograms/ml)-induced slow contractions. These results demonstrated that PFG is a depolarizing neuromuscular blocking agent, being similar to succinylcholine, except that PFG did not produce any contraction, but succinylcholine did.

Acetylcholine↗
PubMed1983

Genes and pseudogenes for mouse U1 and U2 small nuclear RNAs.

Regions of the mouse genome homologous to U1 and U2 small nuclear RNAs have been isolated. The U1 regions are identified as pseudogenes on the basis of: 1) insertions, deletions, and substitutions in the sequence of the DNA as compared with that of U1 RNA; 2) absence of complementary RNA in mouse tissues; and 3) lack of expression upon microinjection in Xenopus oocyte nuclei. A U2 region is identified as a gene on the basis of near identity of sequence with U2 RNA and capacity to direct the synthesis of U2-like RNA in Xenopus oocyte nuclei. Both U1 and U2 sequences are repeated in the mouse genome, the U2 sequences analyzed here in an inverted repeat arrangement.

Animals↗
PubMed1983

Close structural resemblance among small nuclear RNAs.

The published primary and secondary structures of small nuclear RNAs (snRNAs) (1-9) were reexamined and a new common model for their secondary structures was proposed. What is surprising is the resemblance of their secondary structures. U1, U2, U4, and U5 snRNAs share a common sequence of A(U)nG (n greater than or equal to 3) at the single-stranded junction between two hairpins. U6 RNA does not have the A(U)nG sequence, however, all of its possible loops have common sequences with those of U1 RNA at exactly the same positions. Other remarkable common sequences are also noted and their possible biological significance is discussed.

Animals↗
PubMed1980

The emergence of Schistosoma japonicum cercariae from Oncomelania quadrasi.

The release of Schistosoma japonicum cercariae from Leytean Oncomelania quadrasi snails was observed under laboratory conditions. Two patterns of emergence were noted. The initial, nonperiodic emergence occurred immediately after submerging the snails in water and was followed by a periodic, diurnal emergence which peaked in the afternoon. The daily cercarial output of the periodic emergence appeared to be affected by exogenous light intensity. Furthermore, there was a cessation or reduction in cercarial output every 3rd or 4th day.

Animals↗
PubMed1980

Serial implantations of larval Schistosoma mansoni from infected to uninfected snails.

The long-term maintenance of Schistosoma mansoni in intermediate snail hosts is described. Snails were infected with one or five miracidia to obtain the initial parasitized liver tissue. The serial implantations were conducted. When the materials for serial implantation were obtained 20 to 60 days after the hosts had begun to release cercariae, the proportion of snails becoming infected was higher than when materials obtained earlier were used. It made no difference whether the initial infection was with one or five miracidia. Our findings suggest the possibility of cloning of unisexual infections in experimental infections.

Animals↗
PubMed1979

Purification and properties of phosphoglycerate kinase from Thermus thermophilus strain HB8.

(1) A glycolytic enzyme, phosphoglycerate kinase [EC 2.7.2.3], was purified from cells of an extreme thermophile, Thermus thermophilus strain HB8. The enzyme was resistant to heat, and no loss of activity was observed after incubation for 10--20 min at 79 degrees C. (2) Catalytic properties such as pH optimum (pH 6--8.5), kinetic parameters (Km=0.28 mM for ATP, 1.79 mM for glycerate 3-phosphate), substrate specificity and inhibitors of the enzyme were investigated and compared with those of phosphoglycerate kinase from other sources. (3) The enzyme protein consists of a single polypeptide chain of molecular weight 44,600. The isoelectric point is 5.0 The amino acid composition of the enzyme was studied. The contents of ordered secondary structures were estimated to be 29% alpha-helix and 11% pleated sheet from the circular dichroic spectrum of the enzyme protein. (4) The fluorescence spectrum of the enzyme protein showed an emission maximum at 320 nm when excited at 280 nm. The quantum yield was 0.19. Tryptophyl fluorescence was not quenched, in contrast to the fluorescence reported for yeast phosphoglycerate kinase.

Amino Acids↗
PubMed1979

Kietics of thermal unfolding and refolding of thermostable phosphoglycerate kinase.

The kinetics of denaturation by guanidine hydrochloride (GuHCl) of a thermostable phosphoglycerate kinase (PGK) extracted from Thermus thermophilus and of yeast PGK at neutral pH were studied by circular dichroism. Denaturation by GuHCl proceeded as a first-order reaction. The activation free energy of the denaturation reactions (delta Gf not identical to ) in the absence of GuHCl was estimated to be 32.7 kcal/mol for T. thermophilus PGK and 27.9 kcal/mol for yeast PGK (at 25 degrees C). Measurements of the rate constants at various temperatures indicated that delta Gf not identical to has maximum values at 29 degrees C for T. thermophilus PGK and at 20 degrees C for yeast PGK, and that the temperature dependences of delta Gf not identical to, delta Hf not identical to, and delta Sf not identical to for T. thermophilus PGK are smaller than those of yeast PGK. Values of delta Sf not identical to for thermal denaturation for both PGK's are approximately 200 e.u.

Calorimetry↗