PubMed Health⌕ Search

Biomedical subjects

J Miyahara

Publications and source records attributed to J Miyahara.

10 recordsLinked to original sources
PubMed2001

Determination of the antigen/epitope that is recognized by human monoclonal antibody CLN-IgG.

Ever since the development of human monoclonal antibody CLN-IgG in 1982, we anticipated the identification of the antigen that is recognized by this antibody. Despite its scarce expression on the cell surface, susceptibility to proteolytic enzymes and adherence to experimental equipment, we finally succeeded in determining the antigen moiety that is recognized by this antibody by means of CLN-IgG conjugated column affinity chromatography, two-dimensional electrophoresis, MALDI-TOF/MS and use of glioblastoma cell line U-251MG. The antigen was found to be vimentin, a cytoskeletal protein, and we succeeded in determining a 79 amino acids sequence of the epitope which turned out to comprise a part of the c2 (coil 2 of the central rod) domain of vimentin.

Amino Acid Sequence↗
PubMed1997

Large-format imaging plate and weissenberg camera for accurate protein crystallographic data collection using synchrotron radiation.

Off-line and on-line protein data-collection systems using an imaging plate as a detector are described and their components reported. The off-line scanner IPR4080 was developed for a large-format imaging plate ;BASIII' of dimensions 400 x 400 mm and 400 x 800 mm. The characteristics of this scanner are a dynamic range of 10(5) photons pixel(-1), low background noise and high sensitivity. A means of reducing electronic noise and a method for finding the origin of the noise are discussed in detail. A dedicated screenless Weissenberg camera matching IPR4080 with synchrotron radiation was developed and installed on beamline BL6B at the Photon Factory. This camera can attach one or two sheets of 400 x 800 mm large-format imaging plate inside the film cassette by evacuation. The positional reproducibility of the imaging plate on the cassette is so good that the data can be processed by batch job. Data of 93% completeness up to 1.6 A resolution were collected on a single axis rotation and the value of R(merge) becomes 4% from a tetragonal lysozyme crystal using a set of two imaging-plate sheets. Comparing two types of imaging plates, the signal-to-noise ratio of the ST-VIP-type imaging plate is 25% better than that of the BASIII-type imaging plate for protein data collection using 1.0 and 0.7 A X-rays. A new on-line protein data-collection system with imaging plates is specially designed to use synchrotron radiation X-rays at maximum efficiency.

Journal Article↗
PubMed1995

Studies on the quantitative autoradiography. II. Radioluminography for quantitative autoradiography of 3H.

The process of obtaining 3H-autoradiographic (ARG) images has been expected for a long time. A few X-ray films with no protection layer are commercially available, however, they do not give a reliable image which can be quantitated and can give good contrast. We tried to fabricate a 3H-type sensor which has no protection layer on a highly sensitive sensor, and called it Imaging plate (IP). The IP is composed of one of the specially designed photo-stimulable phosphors containing of BaFX: Eu2+(X = Cl, Br or I) crystals. Results indicated a good contrast and reliable whole-body ARG image of 3H-labelled glucose with trial IP, which has never been obtained with any X-ray films even if these were subjected to a long exposure time. The ARG image can be displayed by either black-and-white hard copy or a colored one with the digital display representing relative intensity of photostimulated luminescence (PSL) without back ground (BG) or relative intensity concentration ((PSL-BG)/S), where S was equivalent to 100 pixels (1 mm2). Similarly to the experimental results of 14C, the linear relationship relative intensity and radioactivity of the 3H standard sources was demonstrated with a very wide range of 10(2) to 10(5) dpm/mg upon the exposure for 7d. Also the relationship between relative intensity and relative exposure (radioactivity x exposure time) was linear within the latitude of relative intensity 10(1) to 10(5) (PSL-BG). The trial IP was particularly effective for the quantitative autoradiography of TLC plates.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1991

Molecular cloning and nucleotide sequencing of the Arthrobacter dextranase gene and its expression in Escherichia coli and Streptococcus sanguis.

A bacterial strain, which assimilated dextran and water-insoluble glucan produced by Streptococcus mutans, was isolated from soil. The bacterium produced and secreted potent dextranase activity, which was identified as Arthrobacter sp. and named CB-8. The dextranase was purified and some enzymatic properties were characterized. The enzyme efficiently decomposed the water-insoluble glucan as well as dextran. A gene library from the bacteria was constructed with Escherichia coli, using plasmid pUC19, and clones producing dextranase activity were selected. Based on the result of nucleotide sequencing analysis, it was deduced that the dextranase was synthesized in CB-8 cells as a polypeptide precursor consisting of 640 amino acid residues, including 49 N-terminal amino acid residues which could be regarded as a signal peptide. In the E. coli transformant, the dextranase activity was detected mostly in the periplasmic space. The gene for the dextranase was introduced into Streptococcus sanguis, using an E. coli-S. sanguis shuttle vector that contained the promoter sequence of a gene for glucosyltransferase derived from a strain of S. mutans. The active dextranase was also expressed and accumulated in S. sanguis cells.

Amino Acid Sequence↗
PubMed1988

Imaging plate illuminates many fields.

The erasable phosphor imaging plate developed for medical radiography has found new uses in the laboratory. X-ray diffraction, protein crystallography and autoradiography have all benefited from this technology transfer from the clinic.

Autoradiography↗
PubMed1988

Application of the "imaging plate" to TEM image recording.

The "imaging plate" is a highly sensitive image recording plate for X-ray radiography, which is coated with photo-stimulable phosphor. The imaging plate is exposed to electrons in a transmission electron microscope. Its fundamental properties (sensitivity, dynamic range and sharpness) have been estimated in detail. Also, the image quality of the imaging plate for some specimens in a transmission electron microscope has been estimated. As a result, it has been ascertained that the imaging plate has superior properties and high practicability as an image recording material in a transmission electron microscope.

Animals↗
PubMed1987

Laser-stimulated luminescence used to measure x-ray diffraction of a contracting striated muscle.

An integrating x-ray area detector that operates on the basis of laser-stimulated luminescence was used in a diffraction study of muscle contraction. The area detector has a dynamic range of 1 to 10(5), a sensitivity about 60 times greater with approximately 1/300 as much fog background as x-ray film. It is erasable and reusable but, like film, can integrate at a practically unlimited counting rate. The high sensitivity and wide dynamic range of the detector resulted in a sufficient reduction in the exposure time to make possible the recording of a clear x-ray diffraction pattern, with up to 2.0-nanometer axial spacing, from a contracting frog skeletal muscle in as little as 10 seconds with synchrotron radiation. During the isometric contraction of the muscle, most of the actin diffraction lines increased in intensity without noticeable changes in their peak positions. Changes also occurred in diffraction intensities from the myosin heads. The results indicate that during contraction the structure of the actin filaments differs from that in the rigor state, suggesting a possible structural change in the actin subunits themselves; the myosin heads during contraction retain the axial periodicity of the myosin filament and become aligned in a more perpendicular manner to the actin filaments.

Actin Cytoskeleton↗
PubMed1985

New computed radiography using scanning laser stimulated luminescence.

This short paper summarizes the basic concept and the technology of a new computed radiographic system which uses an energy-storage phosphorus panel called "Imaging Plate" as an image sensor. The "Imaging Plate" can be used to obtain radiographs in exactly the same way as the screen-film combination is used in conventional radiography. The system eliminates the drawbacks of conventional screen-film radiography by combining digital image processing and digitization of the x-ray energy pattern utilizing scanning laser stimulated luminescence.

Analog-Digital Conversion↗
PubMed1983

Computed radiography utilizing scanning laser stimulated luminescence.

A new system of computed radiography that is based on new concepts and the latest computer technologies has been developed. This system eliminates the drawbacks of conventional screen-film radiography. The basic principle of the system is the conversion of the x-ray energy pattern into digital signals utilizing scanning laser stimulated luminescence (SLSL).

Computers↗