Search PubMed⌕ Search

Biomedical subjects

S Yamazaki

Publications and source records attributed to S Yamazaki.

At least 541 records · Page 30Linked to original sources
PubMed1984

Evolution of enterovirus type 70: oligonucleotide mapping analysis of RNA genome.

Different isolates of enterovirus type 70 (EV70) taken between 1971 and 1981 were studied by molecular biological methods to elucidate their evolutional change. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that only a few isolates had a slight alteration in mobility in some viral proteins. On the contrary, oligonucleotide mapping of virion RNA could clearly delineate the molecular changes among isolates. The number of base changes of isolates became greater as the years elapsed. In addition, the number of base changes among recent isolates from different areas of the world was much greater than those among early isolates. Thus, when the isolates were arranged three-dimensionally according to the number of base changes between each other, the constellation of the strains gave rise to a conical shape. The central axis of the figure was the time of isolation of the strains. The early isolates clustered near the top of the conical figure and the recent isolates tended to disperse divergently at the bottom. The figure indicated that all EV70 strains were derived from a common ancestor, and its top would be the time of emergence of the original strain. It was estimated to be around 1966, 3 years prior to the first epidemic in Accra, Ghana. From the results, it was presumed that EV70 would have emerged at a single focus in Africa as a novel human virus. After having circulated there for a few years, the virus spread to the other parts of the world. Based on the difference in the oligonucleotide spots between the recent isolates and early isolates, the base changes of EV70 that occurred during 10 years was estimated to be 320, about 4% (0.4% a year on the average) of the bases of the total RNA genome.

Biological Evolution↗
PubMed1984

Segmental assessment on ordinary scintigrams and SPECT images of the liver.

By examining the area of defect elucidated on the scintigrams and SPECT (single photon emission computed tomography) images, the extension of the tumor into segments was assessed in cases where the SOL (space-occupying lesion) was determined by other methods. In this study, we followed the Couinaud 's segmentation. As a result, it was found that the segment with a SOL could be diagnosed fairly well on radionuclide images. Some incorrect assumptions were also found about the segments on ordinary scintigrams in previously published articles. These incorrect assumptions have been accepted for a long time. Although we do not deny that TCT (transmission X-ray computed tomography), echography, angiography, etc. are necessary for a precise judgment on liver segment and also that there are some cases in which the segmental assessment of these radionuclide images is impossible, we do believe that this kind of effort will improve the diagnostic capability of the radionuclide images.

Adult↗
PubMed1984

Brain extract induces the tetrodotoxin-sensitive action potentials in a rat skeletal muscle cell line (L6).

Tetrodotoxin (TTX) usually blocks the action potential in adult vertebrate innervated skeletal muscle, i.e. the action potential is operated by a TTX-sensitive sodium channel. The L6 cultured myotube, however, has TTX-resistant action potential that is usually found in skeletal muscle following denervation or during fetal and neonatal stages in rat. The TTX-resistivity of the L6 myotubes might be due to a lack of a neurotrophic substance. The present study revealed that brain extract had a trophic substance that is capable of inducing the TTX-sensitive sodium channels in uninnervated cultured L6 myotubes. Seventeen to 18 days old cultures were used and the maximum rate of rise of action potential was measured as an index of the sodium channel density. In control cultures, the maximum rate of rise was about 120 V/s and did not change by TTX application. However, in cultures with the brain extract the value was 142 V/s and reduced to 86 V/s by TTX. The reduced maximum rate of rise by TTX in the latter cultures means that exposure of L6 myotubes to brain extract resulted in an increase in TTX-sensitive sodium channels and a decrease in TTX-resistant sodium channels. Therefore, exposure to a factor in brain extract can elicit development of TTX-sensitive sodium channels in isolated L6 muscle cells that have been grown as a cell line in the absence of neurons for over 10 years.

Action Potentials↗
PubMed1984

Expression of normal and transforming H-ras genes in Escherichia coli and purification of their encoded p21 proteins.

The H-ras gene of the BALB murine sarcoma virus (BALB-MSV) was placed under the transcriptional control of the tightly regulated PL promoter of bacteriophage lambda in the expression vectors pEV-vrf-1 and pRC23. Upon derepression of the PL promoter, large amounts (10-20% of total cellular protein) of the H-ras gene product p21 are synthesized in Escherichia coli. We constructed three H-ras gene expression vectors, designated pJCL-H5, pJCL-E30, and pJCL-33. pJCL-H5 directs the synthesis of p21, a fusion protein whose four amino-terminal residues are replaced by eight amino acids coded for by plasmid sequences. The 13 5' coding nucleotides of the BALB-MSV H-ras gene missing in pJCL-H5 were regenerated in pJCL-E30 by inserting a pair of complementary synthetic oligodeoxynucleotides. As a result, pJCL-E30 encodes a p21 protein, p21T, of sequence identical to that of the transforming p21 protein of BALB-MSV. pJCL-33 is a derivative of pJCL-E30 in which the 12th codon, AAA, a lysine codon, was replaced by GGA, a glycine codon. Thus, pJCL-33 directs the synthesis of a p21 protein, p21N, whose sequence corresponds to that of a normal cellular p21 protein. We report the purification of H-ras p21 proteins to apparent homogeneity by a method involving solubilization with chaotropic agents followed by reverse-phase high-performance liquid chromatography.

Animals↗
PubMed1984

Guanosine nucleotide binding by highly purified Ha-ras-encoded p21 protein produced in Escherichia coli.

High-level expression of the p21 protein product of the BALB murine sarcoma virus v-ras gene (similar to the product of the Harvey murine sarcoma virus v-Ha-ras gene) has been reported recently, and highly purified preparations of this protein have been obtained. We used a nitrocellulose filter assay for measuring the binding of GDP and GTP to the purified protein. Previously p21 antibodies had been used to precipitate p21-guanosine nucleotide complexes from crude extracts containing the protein. Using the filter assay, we find that the v-Ha-ras gene product binds [3H]GDP stoichiometrically. The binding is time-dependent and is faster at 30 degrees C than at 0 degrees C. Optimum binding is obtained in the presence of dithiothreitol and magnesium ions and at pH 7.4. In terms of its GDP binding activity, p21 is heat stable and pronase sensitive. The dissociation constants (Kd) of p21 for [3H]GDP and [3H]GTP, determined by Scatchard analysis, are 6 X 10(-8) M and 2.5 X 10(-8) M, respectively.

Cloning, Molecular↗
PubMed1984

Biological and biochemical characterization of macrophage activating factor (MAF) in murine lymphocytes: physiocochemical similarity of MAF to gamma interferon (IFN-gamma).

During the course of an investigation designed to separate macrophage activating factor (MAF) activity from interferon (IFN) antiviral activity in the lymphokine-rich fraction (LKF) produced by stimulation of murine splenic cells with concanavalin A (Con A), we found molecular evidence for the similarity of the two activities. MAF activity was expressed as the rate of inhibition of intracellular growth of Salmonella typhimurium in macrophages based on the linear correlation between relative MAF activity and LKF concentration. The antiviral substance in LKF was identified as IFN-gamma based on the observation that its activity was inactivated at pH 2 and neutralized with anti-mouse IFN-gamma serum but not with anti-mouse IFN-alpha/beta serum. MAF and IFN antiviral activities displayed identical sensitivity to pH 2 and temperature. Further, neither activity was affected by beta-mercaptoethanol, but both were inactivated by guanidine hydrochloride and by sodium dodecyl sulfate, suggesting that the structures related to conformation of the protein of the two molecules may be similar. In affinity chromatography of the LKF on a Con A-Sepharose column, MAF and IFN activities were found in both the nonadsorbing (F I) and adsorbing (F II) fractions. However, the rates of F II of MAF and IFN activities increased proportionally when the sample was applied on a column of higher capacity, suggesting that the molecular structure of the mannose-containing glycosyl moiety of the two molecules may also be similar. Moreover, the intact or modified form of MAF and IFN activities of different LKF preparations showed a strong correlation, indicating that the production and denaturation of MAF activity were proportional to those of IFN antiviral activity. The results of this study provide strong evidence that MAF and IFN antiviral activities may reside in virtually the same molecular species.

Animals↗
PubMed1984

Characterization of a temperature-sensitive defect of enterovirus 70: effect of elevated temperature on in vitro transcription.

A crude replication complex prepared from enterovirus 70-infected cells was used to study the temperature-sensitive characteristic of the virus. The complex showed a temperature sensitivity in the in vitro incorporation of radiolabeled ribonucleoside triphosphate. The endonuclease itself did not account for the restricted RNA synthesis at the nonpermissive temperature. Analyses of the in vitro products by both gel electrophoresis and sucrose density gradient centrifugation showed that the complex synthesized three types of viral RNA only when incubated for a short period of time at the nonpermissive temperature. When the replication complex was treated with a detergent (deoxycholic acid), incorporation of ribonucleoside triphosphate into RNA at the permissive temperature was reduced to the level of that at the nonpermissive temperature. In addition, the in vitro RNA synthesis by the enterovirus 70 replication complex at the permissive temperature required a higher concentration of ATP than of other ribonucleoside triphosphates, whereas such a preference for ATP was not found in the reaction at the nonpermissive temperature. The results indicate that the initiation step of RNA synthesis by the complex is blocked at the nonpermissive temperature. The possible implications of these findings are discussed.

Adenosine Triphosphate↗
PubMed1984

Primary Budd-Chiari syndrome: ultrasonic demonstration.

Three cases of primary Budd-Chiari syndrome were found by ultrasonic examination. These were confirmed by hepatic venography and inferior vena cavography. The ultrasound findings in these patients included communicating vessels between hepatic veins, enlarged inferior right hepatic vein, reversed blood flow in the hepatic vein, and obstruction of the inferior vena cava. With these findings, ultrasound can be used to diagnose primary Budd-Chiari syndrome without hepatic venography or inferior cavography.

Aged↗
PubMed1984

Genetic relationship between two enteroviruses causing the acute hemorrhagic conjunctivitis syndrome.

The genetic relationship between two enteroviruses causing the acute hemorrhagic conjunctivitis (AHC) syndrome was examined by the RNA-RNA hybridization technique. Nucleotide sequence homology between the J670/71 strain of enterovirus type 70 (EV70) and the EH 24/70 strain of Singapore epidemic conjunctivitis (SEC) virus was found to be only 2% or less of the total RNA genome, while more than 90% homology was detected between the prototype (J670/71) and other isolates of EV70. The results indicate that the SEC virus, an agent of conjunctivitis epidemics in limited areas of southeast Asia and India since 1970, was distinguished serologically and genetically from EV70, which caused pandemics of AHC over the African and Eurasian continents from 1969 to 1972.

Acute Disease↗