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Biomedical subjects

K Nishikawa

Publications and source records attributed to K Nishikawa.

At least 325 records · Page 18Linked to original sources

Protein structural similarities predicted by a sequence-structure compatibility method.

A method for protein structure prediction has been developed, which evaluates the compatibility of an amino acid sequence with known 3-dimensional structures and identifies the most likely structure. The method was applied to a large number of sequences in a database, and the structures of the following proteins were predicted: (1) shikimate kinase (SKase), (2) the hydrophilic subunit of mannose permease (IIABMan), (3) rat tyrosine aminotransferase (Tyr AT), and (4) threonine dehydratase (TDH). The functional and evolutionary implications of the predictions are discussed. (1) The structural similarity between SKase and adenylate kinase was predicted. Alignment of their sequences reveals that the ATP-binding type A sequence motif and 2 ATP-binding arginine residues are conserved. The prediction suggests a similarity in their functional mechanisms as well as an evolutionary relationship. (2) The structural similarity between IIABMan and galactose/glucose-binding protein (GGBP) was predicted. The IIA and IIB domains are aligned with the N- and C-terminal domains of GGBP, respectively. The 2 phosphorylated residues, His 10 and His 175, of IIABMan are threaded onto loops located in the substrate-binding cleft of GGBP. The prediction accounts for the phosphoryl transfer from His 10 to His 175, and to the sugar substrate. (3) The structural similarity between rat Tyr AT and Escherichia coli aspartate AT was predicted, as well as (4) the structural similarity between TDH and the tryptophan synthase beta subunit. Predictions (3) and (4) support the previous predictions based on observations of the functional similarities between the proteins.

Amino Acid Sequence↗

Differences in plaque size and VP4 sequence found in SA11 virus clones having simian authentic VP4.

We isolated five stable SA11 clones (TN-S1, TN-S2, TN-L1, TN-L2, and BN-S4) with different plaque sizes from two SA11 stocks. In polyacrylamide gel electrophoresis, the mobilities of the fourth, fifth, and seventh RNA segments of SA11 clones with large plaque size (TN-L1 and TN-L2) were faster than those of clones with small plaque size (TN-S1, TN-S2, and BN-S4). Nucleotide sequence determination of the fourth RNA segment identified a five-amino-acid difference in VP4s between the clones with large and small plaque sizes. The VP4 sequences of two clones with small plaque size (TN-S1 and BN-S4) were the same as the sequence of SA11-SEM reported by K. Nishikawa et al. (1988, J. Virol. 62, 4022-4026), while the VP4 sequences of the clones with large plaque size (TN-L1 and TN-L2) were similar, but not identical, to that reported by D. B. Mitchell and G. W. Both (1989, Nucleic Acids Res., 17, 2122). A single-gene reassortant, K8-L2.4, in which RNA segment 4 was derived from clone TN-L2 of SA11 virus and the other RNA segments were from strain K8, produced large plaques like TN-L2, suggesting that the five-amino-acid difference in VP4 between the clones with large and small plaque sizes might be associated with the difference in plaque size.

Animals↗

Sequencing and characterization of the kinesin-related genes katB and katC of Arabidopsis thaliana.

Complementary DNAs of two kinesin-related genes, katB and katC, were isolated from Arabidopsis thaliana and sequenced. The carboxyl-terminal regions of the polypeptides encoded by these genes, especially the presumptive ATP-binding and microtubule-binding domains, share significant sequence homology with the mechanochemical motor domain of the kinesin heavy chain. The predicted secondary structures of KatB and KatC proteins include a large globular domain in the carboxyl-terminal region and a small globular domain in the amino-terminal region that are separated by a long alpha-helical coiled-coil with heptad repeats. A truncated KatC polypeptide (KatC(207-754)), which includes the carboxyl-terminal region of KatC, was expressed in Escherichia coli and was shown to possess microtubule-stimulated ATPase activity and to bind to microtubules in an ATP-sensitive manner, both of which are characteristics of kinesin and kinesin-like proteins.

Adenosine Triphosphatases↗

Relationship among coelacanths, lungfishes, and tetrapods: a phylogenetic analysis based on mitochondrial cytochrome oxidase I gene sequences.

To clarify the relationship among coelacanths, lungfishes, and tetrapods, the amino acid sequences deduced from the nucleotide sequences of mitochondrial cytochrome oxidase subunit I (COI) genes were compared. The phylogenetic tree of these animals, including the coelacanth Latimeria chalumnae and the lungfish Lepidosiren paradoxa, was inferred by several methods. These analyses consistently indicate a coelacanth/lungfish clade, to which little attention has been paid by previous authors with the exception of some morphologists. Overall evidence of other mitochondrial genes reported previously and the results of this study equally support the coelacanth/lungfish and lungfish/tetrapod clades, ruling out the coelacanth/tetrapod clade.

Amino Acid Sequence↗

A study of CD45RO, CD45RA and CD29 antigen expression on human decidual T cells in an early stage of pregnancy.

The decidua is the place where the fertilized egg is implanted and where the immunocompetent cells of the mother come into direct contact with genetically disparate cells of the conceptus. Although the T cells in the decidua are exposed to fetal antigens, the fetus is not rejected by maternal immunocompetent cells. In the present study, we examined surface markers to determine whether the T cells in the human decidua are naive T cells without or memory T cells with a history of antigen stimulation. Although few T cells were present in the decidua, as compared to the peripheral blood, CD45RO+, CD29+ and CD45RA- CD4+ T cells as well as CD45RO+, CD29+ and CD45RA- CD8+ T cells, which are considered to be memory T cells, were in the majority, with only small numbers of CD45RO-, CD29- and CD45RA+ CD4+ and CD8+ cells, which are naive T cells, present. Also, the decidual mononuclear cells secreted IL-2 and IL-4. Since IL-4 is secreted only by memory T cells, it is suggested that in the decidua memory T cells increase in number and secrete cytokines, thereby in some way influencing the phenomenon of fertility.

Antigens, CD↗

Miniature endplate currents at the newly formed neuromuscular junction in Drosophila embryos and larvae.

Miniature endplate currents (mepcs) were recorded in embryos and larvae of Drosophila melanogaster with the patch clamp technique in the whole cell configuration from abdominal ventral oblique muscles. Shortly after nerve-muscle contacts (16 h after egg laying) mepcs were small and infrequent. The mean amplitude of mepcs increased abruptly during late embryonic stages but did not change any further up to second instar larvae. The amplitude distribution of mepcs was skewed toward larger amplitudes and changed drastically to a more broader distribution at the time of hatching, but did not change any further. These muscle cells were found to be electrically coupled to those in the rostral neighbor segment. The synaptic current has a fast rising phase and decayed with a single exponential throughout the developmental period examined. The decay time constant increased with amplitude but the rise time did not. Neither the rise time nor the decay time constant changed with development. Many synaptic currents decayed in a stepwise manner revealing underlying single channel events. Two discrete steps were discerned, with amplitudes of about 7.6 and 16 pA at -60 mV. These two types of channels were observed throughout the stages examined.

Animals↗

Basic fibroblast growth factor and steroid receptors in the aging hippocampus of the brown Norway rat: immunocytochemical analysis in combination with stereology.

The effect of aging on the hippocampal formation of the male Brown Norway rat was studied by immunohistochemistry and measurements of the immunoreactive hippocampal cells using stereological techniques. The total estimated number of glucocorticoid receptor (GR) immunoreactive neurons of the CA1-CA2 area did not differ in the 3- and the 36-month-old rat. However, the intensity of the GR immunoreactivity was decreased in the aged animals. A gradual decrease of the immunoreactivity for the mineralocorticoid receptor was also observed in the CA1-CA2 area. In the stratum oriens and the stratum radiatum of the CA1-CA2 area the immunoreactivity for basic fibroblast growth factor (bFGF) present in the glia was found to be reduced [20,000 +/- 2100 (n = 6)] in the 36-month-old rat vs the 3-month-old rat [28,500 +/- 4500 (n = 4) (*P = 0.05)]. However, there was no difference in the number of glial fibrillary acidic protein immunoreactive cells of this area in these two age groups. The present findings give evidence that in the Brown Norway rat there is no loss of the neuronal population containing glucocorticoid receptors of the CA1-CA2 area during aging but suggest that aging is characterized by deficits of glially derived growth factors, such as bFGF.

Aging↗

Unique structure of new serine tRNAs responsible for decoding leucine codon CUG in various Candida species and their putative ancestral tRNA genes.

In an asporogenic yeast, Candida cylindracea, codon CUG is not translated as leucine but as serine. On the basis of our recent work on the determination of the genetic code using in vitro translation systems coupled with isolation of the corresponding tRNA molecules, it appears that this non-universal genetic code is unitized not only in C cylindracea but also in various Hemiascomycetes. Here we show that in addition to the species already reported, three pathogenic yeasts, C guilliermondii, C lusitaniae and C tropicalis, have tRNA(Ser)CAG, indicating that this non-universal genetic code (CUG=Ser) also exists in these species. Determination of their primary structures revealed that the uridine conserved at position 33 in usual tRNAs, is replaced by guanosine or cytidine. This suggests that the three-dimensional structures of the anticodon loop of these tRNAs differ from the conventional structure comprising the U turn in this position. Moreover, we succeeded in isolating putative ancestral serine tRNA genes whose sequences are highly homologous to tRNA(Ser)CAG in each case. These tRNA genes all have the anticodon sequence CGA corresponding to the codon UCG, indicating that tRNA(Ser)CAG might have emerged from tRNA(Ser)CGA during evolutionary change of the assignment of codon CUG.

Base Sequence↗

KP-10, a novel protein kinase C substrate in intact mouse epidermal cells, is phosphorylated by novel protein kinase C eta and/or zeta.

Recently this group found an endogenous substrate protein for Ca(2+)-independent novel protein kinase C (nPKC), i.e. KP-10 (pI 4.7/25,500 M(r)), in primary cultured mouse epidermal cells [Nishikawa, K. et al. (1992) Cell. Signal. 4, 757-776]. In the present study, the nPKC isozymes which phosphorylate KP-10 in these cells were determined. Western blot analysis revealed that PKC alpha, eta and zeta were present in epidermal cell 105,000 g supernatants and that the content of PKC zeta was much higher than those of PKC alpha and eta. Neither PKC beta, delta nor epsilon was detected in the 105,000 g supernatants. Phosphatidylserine and phorbol 12-myristate 13-acetate (PMA)-dependent KP-10 phosphorylating activity was immunoprecipitated by anti-PKC eta and zeta antibodies, but not by antiPKC alpha antibody. These results suggest that PKC eta and/or zeta phosphorylate KP-10 and play pivotal roles in intracellular signal pathways in intact epidermal cells.

Amino Acid Sequence↗

Effects of unusual polyamines on phenylalanyl-tRNA formation.

The effects of novel polyamines on aminoacyl-tRNA formation catalyzed by Escherichia coli., Sulfolobus acidocaldarius, and Thermus thermophilus HB8 S-100 extracts were investigated. These effects were diverse and differed depending on the amino acid and the enzyme used. A quaternary polyamine, tetrakis(3-aminopropyl)ammonium, inhibited phenylalanyl-tRNA synthesis catalyzed by the T. thermophilus extract, but not inhibit the other aminoacyl-tRNA formations tested. The inhibition was observed in hybrid reactions where the thermophile tRNA or extract was replaced by its E. coli counterpart, although the quaternary amine did not inhibit Phe-tRNA formation by the E. coli homologous system. Spermine relieved the inhibition of the reaction of thermophile enzyme and tRNA, but not the inhibition of the hybrid reactions. These results suggest that the branched polyamine interacts with both the thermophile enzyme and tRNA(Phe).

Biogenic Polyamines↗

Bullous eruption of systemic lupus erythematosus.

A bullous eruption is now recognized as a specific complication of systemic lupus erythematosus (SLE). We described a 22-year-old woman with SLE whose vesiculobullous eruptions exacerbated with a flare of visceral manifestations of the disease and resolved when the systemic disease was controlled by oral prednisolone.

Adult↗

Effects of intravenous lidocaine on cardiac sympathetic nerve activity and A-V conduction in halothane-anesthetized cats.

To study neural contributions to the alterations in intracardiac conduction induced by i.v. lidocaine, we measured cardiac sympathetic nerve activity (CSNA) simultaneously with sinus cycle length (SCL) and A-V cats. Sixteen cats were anesthetized with halothane in oxygen and mid-sternotomized. The His-bundle electrogram and CSNA were recorded from an electrode placed in the interatrial septum and from the left ventrolateral or ventromedial nerve, respectively. Atrium-His (A-H), His-Purkinje (H-V), and total intraventricular (H-S) conduction times were measured during atrial pacing conducted at a cycle length of 300 ms. In eight cats, 1 MAC, 2 MAC, and 3 MAC halothane were given during i.v. lidocaine (Groups H-1, H-2 and H-3). In the other eight cats, anesthesia was switched from halothane to i.v. alpha-chloralose (30-50 mg.kg BW-1; Group C). A significant decrease in CSNA with i.v. lidocaine, 2 mg.kg BW-1 was recognized in Groups C and H-1, but not in Groups H-2 and H-3. Prolongations of SCL during the spontaneous cycle, A-H and H-V in the paced mode following i.v. lidocaine were significant in Groups C, H-1 and H-2, but not significant in Group H-3. We conclude that i.v. lidocaine induces a significant decrease in CSNA during alpha-chloralose or 1 MAC halothane anesthesia which partly contributes to the control of intracardiac conduction.

Anesthesia, Inhalation↗

Synthesis and platelet-activating factor (PAF)-antagonistic activities of 1,4-disubstituted piperazine derivatives.

During the screening of novel platelet-activating factor (PAF) antagonists, we found that 1-(6-methoxy-3,4-dihydro-2-naphthoyl)-4- (3,4,5-trimethoxybenzyl)piperazine and its 4-(3,4,5- trimethoxybenzoyl)piperazine derivatives (1b, 2b) exerted in vitro and in vivo PAF-antagonistic activities. Modifications of the 1-acyl group, the substituent at the 4-position and the piperazine ring of 1a and 2b were examined and from this series 1-(2,3-dimethoxy-6,7-dihydro-5H-benzocyclohepten-8-ylcarbonyl++ +)-4- (3,4,5-trimethoxybenzoyl)piperazine (2g) was found to be one of the most potent PAF antagonists.

Animals↗

Synthesis and platelet-activating factor (PAF)-antagonistic activities of trisubstituted piperazine derivatives.

2- or 3-Substituted 1-(2,3-dimethoxy-6,7-dihydro-5H-benzocyclohepten-8- ylcarbonyl)-4-(3,4,5-trimethoxybenzoyl)- and 4-(3,4,5-trimethoxybenzyl)piperazines (2a-s, 3a, b) were prepared and evaluated for antagonistic activities against platelet-activating factor (PAF)-induced platelet aggregation and blood pressure reduction. The 2-methoxymethyl derivative (2f) showed the most potent activities in this series. The enantiomers (R)-(+)-2f and (S)-(-)-2f were synthesized from carbobenzoxy-O-benzyl-L- and D-serine in several steps. In the binding experiment, (S)-(-)-2f showed thirty times greater affinity than the R isomer for the PAF receptor.

Animals↗

Synthesis and angiotensin converting enzyme-inhibitory activity of N-[(1S)-1-carboxy-5-(4-piperidyl)pentyl]-L-alanine derivatives.

As part of a search for potent and long-lasting angiotensin converting enzyme (ACE) inhibitors, various types of N-[(1S)-1-carboxy-5-(4-piperidyl)pentyl]-L-alanine derivatives (7a, 8-11) were prepared. The key synthetic intermediate, N-[(1S)-5-(1-benzyloxycarbonyl-4-piperidyl)-1- ethoxycarbonylpentyl]-L-alanine (17a), was synthesized by asymmetric reduction of the alpha-oxoester (13) with Lactobacillus paracasei subsp. paracasei followed by a substitution reaction with tert-butyl L-alaninate (15) and subsequent treatment with hydrogen chloride. Compounds 7a and 8-11 showed potent and long-lasting ACE-inhibitory activity in rats.

Alanine↗

[Nonpeptide angiotensin II-receptor antagonists].

The renin-angiotensin system (RAS) is known to play a very important role in cardiovascular diseases. The present state of the research on orally active nonpeptide angiotensin II (A II)-receptor antagonists and their pharmacological characterization will be reviewed. In the late 1970s, the first nonpeptide A II-receptor antagonists were discovered among some derivatives of imidazoleacetic acid, and this was followed by the development of losartan in 1989. TCV-116, synthesized in our laboratories, is a prodrug that is converted in vivo to the active form CV-11974. TCV-116 and CV-11974 were demonstrated to be effective antagonists for many A II-induced cardiovascular actions and are effective antihypertensive agents in several animal models of hypertension. TCV-116 also demonstrated secondary benefits in treating congestive heart failure (CHF), preventing stroke, delaying the progression of renal disease and preventing the intimal thickening of vascular injury in animal models. Clinical studies confirmed the efficacy of TCV-116 in the treatment of essential hypertension. The utility of A II antagonist may extend beyond that of hypertension and CHF, as suggested by the potential usefulness of ACE inhibitors in the treatment or prevention of many other cardiovascular diseases. The A II antagonists will help to determine the role of the RAS in the physiologic regulation and in the pathophysiology of various cardiovascular diseases.

Angiotensin Receptor Antagonists↗

Quantitative assessment of image quality using niobium filtration for cephalometric radiography.

The effect of added niobium filtration on image quality in cephalometric radiography was quantitatively evaluated using the Modulation Transfer Function (MTF). A conventional full-wave cephalometric X-ray generator (Quint Sectograph) and a constant potential, high frequency X-ray generator (Orthophos CD) were compared with respect to the effects on the MTF of the added filtration. There was a slight reduction in image quality with both machines when adding niobium filtration.

Cephalometry↗