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Masahiro Kajiwara

Publications and source records attributed to Masahiro Kajiwara.

11 recordsLinked to original sources

Ligand-induced structural changes of the CD44 hyaluronan-binding domain revealed by NMR.

CD44, a major cell surface receptor for hyaluronan (HA), contains a functional domain responsible for HA binding at its N terminus (residues 21-178). Accumulating evidence indicates that proteolytic cleavage of CD44 in its extracellular region (residues 21-268) leads to enhanced tumor cell migration and invasion. Hence, understanding the mechanisms underlying the CD44 proteolytic cleavage is important for understanding the mechanism of CD44-mediated tumor progression. Here we present the NMR structure of the HA-binding domain of CD44 in its HA-bound state. The structure is composed of the Link module (residues 32-124) and an extended lobe (residues 21-31 and 125-152). Interestingly, a comparison of its unbound and HA-bound structures revealed that rearrangement of the beta-strands in the extended lobe (residues 143-148) and disorder of the structure in the following C-terminal region (residues 153-169) occurred upon HA binding, which is consistent with the results of trypsin proteolysis studies of the CD44 HA-binding domain. The order-to-disorder transition of the C-terminal region by HA binding may be involved in the CD44-mediated cell migration.

Amino Acid Sequence↗

Proposal for internet-based Digital Dental Chart for personal dental identification in forensics.

A dental chart is very useful as a standard source of evidence in the personal identification of bodies. However, the kind of dental chart available will often vary as a number of types of odontogram have been developed where the visual representation of dental conditions has relied on hand-drawn representation. We propose the Digital Dental Chart (DDC) as a new style of dental chart, especially for open investigations aimed at establishing the identity of unknown bodies. Each DDC is constructed using actual oral digital images and dental data, and is easy to upload onto an Internet website. The DDC is a more useful forensic resource than the standard types of dental chart in current use as it has several advantages, among which are its ability to carry a large volume of information and reproduce dental conditions clearly and in detail on a cost-effective basis.

Forensic Dentistry↗

In vivo studies of phenylalanine hydroxylase by phenylalanine breath test: diagnosis of tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency.

Tetrahydrobiopterin (BH4)-responsive phenylalanine hydroxylase (PAH) deficiency is characterized by reduction of blood phenylalanine level after a BH4-loading test. Most cases of BH4-responsive PAH deficiency include mild phenylketonuria (PKU) or mild hyperphenylalaninemia (HPA), but not all patients with mild PKU respond to BH4. We performed the phenylalanine breath test as reliable method to determine the BH4 responsiveness. Phenylalanine breath test quantitatively measures the conversion of L-[1-13C] phenylalanine to 13CO2 and is a noninvasive and rapid test. Twenty Japanese patients with HPA were examined with a dose of 10 mg/kg of 13C-phenylalanine with or without a dose of 10 mg . kg(-1) . d(-1) of BH4 for 3 d. The phenylalanine breath test [cumulative recovery rate (CRR)] could distinguish control subjects (15.4 +/- 1.5%); heterozygotes (10.3 +/- 1.0%); and mild HPA (2.74%), mild PKU (1.13 +/- 0.14%), and classical PKU patients (0.29 +/- 0.14%). The genotypes in mild PKU cases were compound heterozygotes with mild (L52S, R241C, R408Q) and severe mutations, whereas a mild HPA case was homozygote of R241C. CRR correlated inversely with pretreatment phenylalanine levels, indicating the gene dosage effects on PKU. BH4 loading increased CRR from 1.13 +/- 0.14 to 2.95 +/- 1.14% (2.6-fold) in mild PKU and from 2.74 to 7.22% (2.6-fold) in mild HPA. A CRR of 5 to 6% reflected maintenance of appropriate serum phenylalanine level. The phenylalanine breath test is useful for the diagnosis of BH4-responsive PAH deficiency and determination of the optimal dosage of BH4 without increasing blood phenylalanine level.

Adolescent↗

[Simple quantitation of arsenic by energy dispersive fluorescence X-ray spectrometer using Reinsch's test].

We examined the clinical usefulness of the Reinsch's test for the detection of the small amounts of the heavy metals such as arsenic and mercury using the fluorescence X-ray spectrometry. We tried t o measure various kinds of biological samples, including serum, urine, and gastric contents using this method. 0.4 ml or 1 ml of hydrochloric acid were added to 2 ml of serum or 6 ml urine and gastric content, respectively, and a copper plate (5.0x 0.8 cm) was immersed into this solution. The mixture heated at 90 degrees C by a heating block for 30 minutes. After heating, the copper plate was washed with water and dried. The copper arsenide that stuck to the copper plate due to Reinsch's test dissolved by methanol/ammonia (8:2) solution at 60 degrees C for 15 minutes. A drop gave the solution to a filter paper fluorescence X-ray analysis and completely dried the filter paper, and applied to the fluorescence X-ray spectrometer. As a result, this method showed about 20 times high sensitivity in comparison with the measurement with condition of solution. The minimal detectable limits of rsenic was 0.4 ppm in serum and were 0.2 ppm in rine and gastric content. The calibration curve could be made for 0.5 to 50 ppm. It will take about 90 min for the measurement using this method for the detection of arsenic in biological samples. We showed the usefulness of the Reinsch's test using the fluorescence X-ray spectrometry in the clinical toxicology.

Acute Disease↗

Hyaluronan recognition mode of CD44 revealed by cross-saturation and chemical shift perturbation experiments.

CD44 is the main cell surface receptor for hyaluronic acid (HA) and contains a functional HA-binding domain (HABD) composed of a Link module with N- and C-terminal extensions. The contact residues of human CD44 HABD for HA have been determined by cross-saturation experiments and mapped on the topology of CD44 HABD, which we elucidated by NMR. The contact residues are distributed in both the consensus fold for the Link module superfamily and the additional structural elements consisting of the flanking regions. Interestingly, the contact residues exhibit small changes in chemical shift upon HA binding. In contrast, the residues with large chemical shift changes are localized in the C-terminal extension and the first alpha-helix and are generally inconsistent with the contact residues. These results suggest that, upon ligand binding, the C-terminal extension and the first alpha-helix undergo significant conformational changes, which may account for the broad ligand specificity of CD44 HABD.

Amino Acid Sequence↗

DNA analysis of neonatal human remains wrapped and kept in a vinyl bag for 15 years.

DNA analysis of a newborn baby wrapped and kept in a vinyl bag for 15 years was performed. DNA isolated from the femur and humerus was used to determine the sex and kinship between the infant and the putative parents. Amplification of mtDNA, ABO, HLA, CST3, CST5, VWA, D12S66, D21S11, CSF1PO, TPOX, THO1 and 10Y polymorphisms and the amelogenin gene was carried out. Several mtDNA types were obtained, suggesting that the sample was contaminated by exogenous DNAs. One of the DNA samples obtained from the femur showed an identical mtDNA sequence to that of the mother except for one site, and this pattern was also found in another DNA sample. None of our laboratory personnel had that type, so we thought it was possible that this sample contained the target DNA. However, maternity was denied by the CST3 polymorphism. Finally, we concluded that the sample had been contaminated with exogenous DNA before we started to examine the body. Although it is difficult to determine the sources of this contamination, PCR amplification from highly degraded DNA is very sensitive to such contamination, and we must be even more careful in DNA analysis of such samples than in that of not so severely degraded specimens.

DNA↗

[13C-urea breath test].

Infection with Helicobacter pylori(H. pylori) is thought to be an important factor in the pathogenesis of gastritis, peptic ulcer and gastric cancer, and the 13C-urea breath test (13C-UBT) is a convenient and non-invasive method for the detection of H. pylori in the stomach. We have examined the sensitivity, specificity and accuracy of 13C-UBT, comparing it with histology. The 13CO2/12CO2 ratio was measured by using infra-red spectroscopy(IR) and gas chromatography/mass spectrometry(GC-MS). The IR analyzer should be particularly useful for the diagnostic test of H. pylori infection using 13C-UBT.

Breath Tests↗

Evaluation of intraoral CCD camera for dental examination in forensic inspection.

This study was performed to assess the effectiveness of an intraoral CCD camera for dental examinations when sufficient jaw opening or adequate lighting cannot be obtained. A handpiece-type intraoral CCD camera (Crystal Cam; GC Corp., Japan) was used for the study. Because a full view taken by the intraoral CCD camera covers only one or two teeth, all the teeth were individually photographed and a view of the dentition assembled on a personal computer. Assuming that the jaw could not be opened widely enough to inspect an occlusal view, a dry skull and a volunteer were restricted to open the mouth and all the teeth were photographed with an intraoral CCD camera. These were compared to intraoral photographs taken by the conventional method using a single-lens reflex camera and mirror. When the intraoral CCD camera was used to photograph teeth, the color tone of metal restorations could be readily identified, but special care was required to identify carious lesions, discoloration of tooth structure, and esthetic restorations. The dentition photographs assembled from the original intraoral CCD images were transferred via the Internet as e-mail attachment files to allow preparation of the dental chart at the destination. Based on the transferred images, it was possible to prepare a dental chart agreeing satisfactorily with actual oral conditions. The easy transfer of digital images provides various advantages in evaluating and discussing certain cases in cooperation with other forensic odontologists via the Internet. The camera may be made more effective or useful through improvement of the tip portion of the camera and the entire system to achieve a more compact design and better portability.

Journal Article↗

Evaluation of two biosynthetic pathways to delta-aminolevulinic acid in Euglena gracilis.

delta-Aminolevulinic acid (ALA), which is an intermediate in the biosynthesis of chlorophyll a, can be biosynthesized via the C5 pathway and the Shemin pathway in Euglena gracilis. Analysis of the (13)C-NMR spectrum of (13)C-labeled methyl pheophorbide a, derived from 13C-labeled chlorophyll a biosynthesized from d-[1-(13)C]glucose by E. gracilis, provided evidence suggesting that ALA incorporated in the (13)C-labeled chlorophyll a was synthesized via both the C5 pathway and the Shemin pathway in a ratio of between 1.5 and 1.7 to one. The methoxyl carbon of the methoxycarbonyl group at C-132 of chlorophyll a was labeled with (13)C. The phytyl moiety of chlorophyll a was labeled on C-P2, C-P3(1), C-P4, C-P6, C-P7(1), C-P8, C-P10, C-P11(1), C-P12, C-P14, C-P15(1) and C-P16.

Aminolevulinic Acid↗