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Biomedical subjects

S Tsuge

Publications and source records attributed to S Tsuge.

At least 19 recordsLinked to original sources
PubMed2001

Quantitative analysis of detailed lignin monomer composition by pyrolysis-gas chromatography combined with preliminary acetylation of the samples.

Detailed quantitative analysis of lignin monomer composition comprising p-coumaryl, coniferyl, and sinapyl alcohol and p-coumaraldehyde, coniferaldehyde, and sinapaldehyde in plant has not been studied from every point mainly because of artifact formation during the lignin isolation procedure, partial loss of the lignin components inherent in the chemical degradative methods, and difficulty in the explanation of the complex spectra generally observed for the lignin components. Here we propose a new method to quantify lignin monomer composition in detail by pyrolysis-gas chromatography (Py-GC) using acetylated lignin samples. The lignin acetylation procedure would contribute to prevent secondary formation of cinnamaldehydes from the corresponding alcohol forms during pyrolysis, which are otherwise unavoidable in conventional Py-GC process to some extent. On the basis of the characteristic peaks on the pyrograms of the acetylated sample, lignin monomer compositions in various dehydrogenative polymers (DHP) as lignin model compounds were determined, taking even minor components such as cinnamaldehydes into consideration. The observed compositions by Py-GC were in good agreement with the supplied lignin monomer contents on DHP synthesis. The new Py-GC method combined with sample preacetylation allowed us an accurate quantitative analysis of detailed lignin monomer composition using a microgram order of extractive-free plant samples.

Acetylation↗
PubMed2001

Stabilization of cauliflower mosaic virus P3 tetramer by covalent linkage.

Cauliflower mosaic virus (CaMV) open reading frame (ORF) III encodes a 15 kDa protein (P3) that is indispensable for viral infectivity. Although P3 has been shown to be a prerequisite for CaMV aphid transmission, its role in viral replication remains unknown. We previously showed that P3 forms a tetramer in planta and that P3 tetramer co-sediments with viral coat protein on sucrose gradient centrifugation, suggesting that a tetramer may be the functional form of P3. We presumed that disulfide bonds were involved in tetramer formation because 1) the tetramer was detected by Western blotting after electrophoresis under non-reducing conditions, and 2) the cysteine-X-cysteine motif is well conserved in CaMV P3 and P3 homologues among Caulimoviruses. Therefore we mutated either or both of the cysteine residues of CaMV P3. The mutant viruses were infectious and accumulated to a similar extent as the wild-type. An analysis of mutant proteins confirmed that the wild-type P3 molecules in the tetramer are covalently bound with one another through disulfide bonds. It was also suggested that mutant proteins are less stable than wild-type protein in planta. Furthermore, sedimentation study suggested that the disulfide bonds are involved in stable association of P3 with CaMV virions or virion-like particles, or both. The mutant viruses could be transmitted by aphids. These results suggested that the covalent bonds in P3 tetramer are dispensable for biological activity of P3 under experimental situations and may have some biological significance in natural infection in the field.

Alanine↗
PubMed2001

Identification of a H+/glucose and galactose symporter gene glt from Xanthomonas oryzae pv. oryzae.

We identified a glucose and galactose transporter gene from the plant-pathogenic bacterium Xanthomonas oryzae pv. oryzae. Sequence analysis indicated that the gene, named glt, encoded a polypeptide of 592 amino acid residues and the product was significantly homologous with members of the Na+/glucose cotransporter (SGLT) family from mammalian and bacterial origin, especially with vSGLT from Vibrio parahaemolyticus (50% identity). GLT functioned as a glucose and galactose transporter in an Escherichia coli mutant deficient in glucose and galactose transport activity. A protonophore inhibited the transport activity, suggesting that GLT is a H+-coupled glucose/galactose symporter.

Amino Acid Sequence↗
PubMed2001

Optimization of conditions for detailed compositional analysis of acrylic oligomers by supercritical fluid chromatography with temperature programming or modifier gradient technique.

Acrylic oligomers such as pentaerythritolacrylates (PEA), which are often utilized in the field of functionalized materials, generally consist of very complex homologous series. In this work, measuring conditions for packed-column supercritical fluid chromatographic separation of a complex PEA sample manufactured industrially were systematically examined using temperature programming or modifier gradient technique. Under the conditions thus optimized, not only the main components, PEA homologues containing different number of acryloyl groups, but also trace constituents such as their by-products formed through Michael addition reactions were favorably resolved on the resulting chromatograms. The fact that the coefficient of variation (CV) for relative intensity of each peak was approximately 3-6% even for the minor components suggested that detailed compositional analysis of PEAs could be performed precisely by the proposed method.

Journal Article↗
PubMed1999

Determination of benzodiazepine in tablets studied by thermal desorption gas chromatography.

Thermal desorption gas chromatography (TDGC) was applied to the analysis of 13 kinds of tablets containing different benzodiazepines (BZDs). An aliquot of ground tablet sample (0.1-1 mg), weighed into a platinum sample cup, was placed in a furnace pyrolyzer where the sample was heated up to a suitable temperature (150-500 degrees C) so that BZD was desorbed from the sample powder. The desorbed components of BZDs were immediately transferred into a separation column by a helium carrier gas without using any trapping techniques. The desorbed components were identified by TDGC-mass spectrometry. Among various BZDs, fludiazepam, nimetazepam and mexazolam in tablet samples were determined by the present method. Thus, the results obtained were in good agreement with the specified values. Correlation coefficients of the calibration lines for the three BZDs' ranged from 0.997 to 0.999 for several micrograms to about 10 microg of the components. Relative standard deviations of this method were < 4.1% for 4 or 5 runs.

Adsorption↗
PubMed1999

Cauliflower mosaic virus ORF III product forms a tetramer in planta: its implication in viral DNA folding during encapsidation.

Cauliflower mosaic virus (CaMV) open reading frame (ORF) III encodes a 15 kDa protein; the function of which is as yet unknown. This protein has non-sequence-specific DNA binding activity and is associated with viral particles, suggesting that the ORF III product (P3) is involved in the folding of CaMV DNA during encapsidation. In this study, we demonstrated that P3 forms a tetramer in CaMV-infected plants. A P3-related protein with an apparent molecular weight of 60 kDa was detected by Western blotting analysis using anti-P3 antiserum under non-reducing conditions, while only 15 kDa P3 was detected under reducing conditions. Analysis of P3 using viable mutants with a 27-bp insertion in either ORF III or IV revealed that the 60 kDa protein was a tetramer of P3. The P3 tetramer co-sedimented with viral coat protein in multiple fractions on sucrose gradient centrifugation, suggesting that P3 tetramer binds to mature and immature virions. These results strongly suggested that CaMV P3 forms a tetramer in planta and that disulfide bonds are involved in its formation and/or stabilization. The finding of P3 tetramer in planta suggested that viral DNA would be folded compactly by the interaction with multiple P3 molecules, which would form tetramers, while being packaged into the capsid shell.

Brassica↗
PubMed1998

Correlation analysis between fatty acid compositions of zooplankter individuals, fed on different phytoplankton species by means of pyrolysis-gas chromatography combined with on-line methylation.

Pyrolysis-gas chromatography (Py-GC) combined with on-line methylation was applied to a correlation analysis between the distributions of fatty acid components in the lipids of zooplankter individuals and those of ingested algae using principal component analysis (PCA). Py-GC in the presence of organic alkali, tetramethylammonium hydroxide (TMAH), was used to estimate the apparent distributions of fatty acid components contained in a single individual zooplankter weighing several tens of micrograms and a small sample size of ingested algae samples in the order of 10 microg. The observed fatty acid compositions were used as a database for the PCA in order to discriminate the zooplankton and ingested algae samples. The result obtained indicated that the fatty acid compositions of zooplankton individuals used in this work were significantly reflected in those of their ingested food in spite of some contribution from isomerization and/or elongation of fatty acid components during digestion of the ingested algae phytoplankton in living zooplankters.

Animals↗
PubMed1998

Accumulation kinetics of viral gene products in cauliflower mosaic virus-infected turnip protoplasts.

The expression of cauliflower mosaic virus (CaMV) genes was studied in a turnip protoplast system. Six CaMV-encoded gene products were detected in infected turnip protoplasts by means of Western blotting. The infected turnip protoplasts showed different patterns of protein accumulation; e.g. an open reading frame (ORF) I-encoded movement protein, an ORF V-encoded reverse transcriptase and an ORF VI-encoded posttranscriptional transactivator representing the early accumulated proteins, an ORF II-encoded aphid transmission factor and an ORF IV-encoded coat protein the late accumulated proteins and an ORF III-encoded DNA binding protein the intermediate protein. The results suggest that the expression of CaMV genes is differentially regulated.

Blotting, Western↗
PubMed1998

Evidence for a dual strategy in the expression of cauliflower mosaic virus open reading frames I and IV.

Studies have indicated that cauliflower mosaic virus (CaMV) gene expression is mediated by the translation of polycistronic 35S pregenomic RNA, but the involvement of some minor subgenomic RNA species is also suspected. We examined the involvement of the 35S promoter in the expression of CaMV open reading frames (ORFs) I and IV using both 35S RNA-driven and promoter-less ORF I- and ORF IV-beta-glucuronidase (GUS) fusion constructs. In addition to the 35S promoter-dependent expression of both ORF I- and IV-GUS fusions, we detected the 35S promoter-independent expression of both fusion genes via subgenomic mRNAs, which were detected by Northern blotting in the protoplasts transfected with the 35S promoter-driven constructs as well as in those transfected with the promoter-less constructs. These results suggest the involvement of subgenomic RNAs in the expression of CaMV ORFs I and IV, and the operation of a dual strategy in the expression of two viral genes.

Caulimovirus↗
PubMed1998

Requirement of cauliflower mosaic virus open reading frame VI product for viral gene expression and multiplication in turnip protoplasts.

Cauliflower mosaic virus (CaMV) open reading frame (ORF) VI product (P6) has been shown to be the major constituent of viral inclusion body, to function as a post-transcriptional transactivator, and to be essential for infectivity on whole plants. Although these findings suggest that P6 has an important role in viral multiplication, it is unknown whether P6 is required for viral multiplication in a single cell. To address this question, we transfected turnip protoplasts with an ORF VI frame-shift (4 bp deletion) mutant (pCaFS6) of an infectious CaMV DNA clone (pCa122). The mutant was uninfectious. Co-transfection of plasmids expressing P6 complemented the mutant. Overexpression of P6 elevated the infection rate in co-transfection experiments with either pCa122 or pCaFS6. This would have been achieved by elevating the level of pregenomic 35S RNA, a putative polycistronic mRNA for ORFs I, II, III, IV and V, and by enhancing the accumulation of these five viral gene products. When CaMV ORFs I, II, III, IV and V were expressed from monocistronic constructs in which each of the ORFs was placed just downstream of the 35S promoter, the accumulation of ORF III, IV and V products depended on the co-expression of P6. The accumulation of ORF I and II products was not detected, even in the presence of P6. These results suggest that P6 is involved in the stabilization of other viral gene products as well as in the activation of viral gene expression, and thus, is a prerequisite for CaMV multiplication.

Blotting, Northern↗
PubMed1998

Analysis of lipid components in zooplankter individuals by reactive pyrolysis-gas chromatography in the presence of organic alkali.

It has been eagerly requested to develop a highly-sensitive method to characterize extremely minute amounts of natural organic materials occluded in meteorites and/or space dusts in order to confirm the existence of life in the extraterrestrial space. In this article, the reactive pyrolysis-gas chromatography (Py-GC) applied to the analysis of the lipid components contained in every zooplankter individual is introduced for the sake of its potential extension to the characterization of trace amounts of the extraterrestrial organic materials. Here, Py-GC was applied to 1) the discriminative analysis among zooplankter individuals cultured in different food concentrations, and 2) the correlation analysis between the distributions of fatty acid components in the lipids of zooplankter individuals and ingested algae phytoplankton.

Animals↗
PubMed1997

A method for determining the distribution of fluoride, calcium and phosphorus in human dental plaque and the effect of a single in vivo fluoride rinse.

A new sampling method, capable of sampling plaque from its surface to its interior for quantitative studies, was modified to meet some of the requirements for the determination of the fluoride and mineral (Ca and P) profiles within dental plaque formed in vivo. Plaque samples were repeatedly collected from the same individual, using special devices, before a single fluoride rinse (900 parts/10(6) fluoride) and 10 min and 24 hr after rinse. The method allowed examination of fluoride, calcium and phosphorus distribution along the entire thickness of plaque. Fluoride content significantly increased throughout the sample 10 min after rinsing, indicating the fluoride had rapidly penetrated into the plaque. Although the elevated fluoride concentrations diminished almost to baseline with 24 hr, a high correlation was found between fluoride and minerals in each plaque fraction. It is concluded that this technique will be useful for evaluating the fluoride and mineral behaviour in the saliva/plaque and plaque/enamel interfaces, and the anti caries efficacy of fluoride applications.

Calcium↗
PubMed1993

Anatomical risk factors for Kienböck's disease.

Contralateral unaffected wrists from 41 males with Kienböck's disease were compared with wrists from 66 normal males. From X-rays, various features of the lunate and radius were measured. In patients with Kienböck's disease, the lunate tended to be smaller and inclined more radially than in normal subjects and the radial inclination was flatter. Discriminant analysis showed that 85% of the unaffected contralateral wrists in patients with Kienböck's disease and 74% of the wrists in normal subjects were accurately discriminated to their respective groups. It may be possible to identify subjects who are at risk for Kienböck's disease prior to onset using discriminant analysis.

Adult↗
PubMed1993

Reasons for the low accumulation level of aphid transmission factor protein in infected leaves with an aphid-non-transmissible cauliflower mosaic virus isolate, CM1841.

The synthesis and accumulation of aphid transmission factor protein (p18) in cauliflower mosaic virus (CaMV)-infected turnip protoplasts were examined in time course and pulse-labelling experiments, comparing an aphid-non-transmissible isolate (CM1841) with an in vitro recombinant aphid-transmissible CaMV (CMBX) generated from the CM1841 isolate. There was little difference in the synthesis and accumulation of p18 between CM1841- and CMBX-infected protoplasts. When the accumulation of p18 in infected leaves was monitored from 3 to 28 days post-symptom emergence (p.e.) by Western blotting, the amount of p18 accumulated in CM1841-infected leaves continuously decreased from 3 days p.e. throughout the experimental period, whereas the amount of p18 in CMBX-infected leaves was lowest at 3 days p.e. and increased thereafter. These results suggested that CM1841 differed from CMBX not in the synthesis of p18 but in the stability of p18 in infected leaves.

Animals↗
PubMed1992

Nucleotide sequence responsible for the synthesis of a truncated coat protein of brome mosaic virus strain ATCC66.

The ATCC66 strain of brome mosaic virus (BMV) (propagated at Kyoto University) contained two types of coat protein in its virion whereas the Russian strain of BMV has been known to contain a single coat protein; both strains have two initiation codons for coat protein in the same reading frame at the 5'-proximal end of the gene in RNA 4. Comparative studies on the nucleotide sequences of the ATCC66 and Russian strains of BMV demonstrated that in the ATCC66 strain, two adjacent adenine residues were absent from RNA 3 in the leader sequences of the coat protein gene just a few nucleotides 5' to the first initiation codon of the coat protein gene. Using biologically active cDNA clones of BMV RNA of the ATCC66 strain, we inserted two adjacent adenine residues into the cDNA of RNA 3 to obtain an RNA 3 transcript which has the same nucleotide sequence as the Russian strain in the non-coding leader sequence of the coat protein gene. Barley protoplasts inoculated with this RNA 3 transcript together with RNA 1 and 2 produced a single coat protein. To obtain further insight into the mechanism of translation of the BMV coat protein, we constructed several types of RNA 4 by changing the sequence surrounding the first AUG codon in the coat protein gene and analysed the in vitro translation products of the mutant RNA 4. The results confirmed that the absence of the two adjacent adenine residues was responsible for the production of two types of coat protein in the ATCC66 strain. The deletion of the two adjacent adenine residues in ATCC66 resulted in a base substitution of A with U three nucleotides 5' to the first AUG in the coat protein gene. The base substitution reduced translational activity from the first AUG codon and concomitantly increased translational activity from the second AUG codon from which a truncated coat protein was translated.

Capsid↗
PubMed1991

Scaphoid non-union with D.I.S.I. deformity. A survey of clinical cases with special reference to ligamentous injury.

To clarify whether or not scapho-lunate ligamentous injury is common in scaphoid non-union with D.I.S.I. deformity and whether ligamentous repair is required to treat scaphoid non-union with D.I.S.I. deformity, roentgenographic, arthrographic and arthroscopic findings were analysed and compared to those of scaphoid non-union without D.I.S.I. deformity. The carpal alignment of scaphoid non-union with D.I.S.I. deformity treated by reduction and anterior wedge grafting without ligamentous repair was also studied. No significant difference in the scapho-lunate gap was present between 26 patients with D.I.S.I. deformity and 20 without. No statistically significant difference was detected in the incidence of arthrographically-proved scapho-lunate ligamentous tears between the two groups. Arthroscopic examination carried out in 22 patients did not detect massive ligamentous injury, and carpal alignment was satisfactory in most patients who underwent anterior wedge grafting. These results show that ligamentous injury rarely causes D.I.S.I. deformity in scaphoid non-union, and anterior wedge grafting alone is sufficient to correct D.I.S.I. deformity in most cases.

Adolescent↗
PubMed1991

Radial wedge osteotomy for Kienböck disease.

A radial wedge osteotomy, with reduction of the inclination angle of the distal part of the radius, was performed in twenty-seven patients who had Kienböck disease. After two to five years of follow-up, all of the patients were free of pain or had only mild pain in the wrist with strenuous activity. Flexion-extension of the wrist improved by more than 10 degrees in about half of the patients, and in most of the patients the grip strength increased by five kilograms or more. All but two of the postoperative results were satisfactory, as evaluated by the criteria of Lichtman et al. The osteotomy was effective even in patients who had zero or positive ulnar variance.

Adolescent↗