[A case of intestinal Behçet disease with duodenal ulcer perforation].
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Behcet Syndrome↗
Biomedical subjects
M Machida
Publications and source records attributed to M Machida.
Detection and mapping of mismatched base pairs in DNA molecules by atomic force microscopy.
Attempts were made to apply atomic force microscopy (AFM) imaging to the detection and mapping of the sites of base substitutions in DNA molecules. In essence, DNA fragments to be examined for possible base substitutions were mixed with an equal amount of a corresponding DNA standard and subjected to heat denaturation and subsequent annealing. The reassociated DNA was incubated with MutS protein, a protein that recognizes and binds to mismatched base pairs in duplex DNA. Bound MutS protein molecules were then detected by AFM and their positions along the DNA molecules were determined by calculating the distance from one of the DNA termini, which had been tagged with a biotin-avidin complex. Base substitutions present in DNA molecules >1 kb were effectively detected by this procedure, and the positions determined were in good agreement with the actual mutation sites. This method is quite simple, has virtually no limitations on the size of DNA fragments to be examined and requires only a very small amount of DNA sample.
Adenosine Triphosphatases↗
Global Simulations of Differentially Rotating Magnetized Disks: Formation of Low-beta Filaments and Structured Coronae.
We present the results of three-dimensional global magnetohydrodynamic simulations of the Parker-shearing instability in a differentially rotating torus initially threaded by toroidal magnetic fields. An equilibrium model of a magnetized torus is adopted as an initial condition. When beta0=Pgas&solm0;Pmag approximately 1 at the initial state, magnetic flux buoyantly escapes from the disk and creates looplike structures similar to those in the solar corona. Inside the torus, the growth of nonaxisymmetric magnetorotational (or Balbus & Hawley) instability generates magnetic turbulence. Magnetic field lines are tangled on a small scale, but on a large scale they show low azimuthal wavenumber spiral structure. After several rotation periods, the system oscillates around a state with beta approximately 5. We found that magnetic pressure-dominated (beta<1) filaments are created in the torus. The volume filling factor of the region in which beta</=0.3 is 2%-10%. Magnetic energy release in such low-beta regions may lead to violent flaring activities in accretion disks and in galactic gas disks.
Journal Article↗
A 38 kb segment containing the cdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment.
A genomic 38 kbp segment on the c1750 cosmid clone containing the cdc2 gene, located in the left arm of chromosome II from Schizosaccharomyces pombe, was sequenced. The segment was found to have five previously known genes, pht1, cdc2, his3, act1 and mei4. Among 11 coding sequences (CDSs) predicted by the gene finding software INTRON.PLOT., four CDSs, pi007, pi010, pi014 and pi016, had considerable similarity to 40S ribosomal protein, glycosyltransferase, cdc2-related protein kinase and alpha-1, 2-mannosyltransferase, respectively. Another unusually huge open reading frame (ORF) (pi011), consisting of 2233 amino acids, existed, having significant homology to alpha-amylase, granule-bound glycogen synthase and the Sz. pombe YS 1110 clone product at the N-terminal, middle and C-terminal regions, respectively. All the predicted 11 CDSs were experimentally analysed by RACE PCR. The sequencing of the RACE products revealed that there were two small overlaps at the 3' untranslated regions (UTRs) between pi004 and pi005 (17 bp) and between pi007 and pi008 (2 bp). The distances between 5' end of the 5'UTR and the putative translation initiation codon varied from 10 to 302 nucleotides (nt) among the nine CDSs successfully analysed by 5'-RACE. The expression level of each CDS on this clone was determined. Among the 16 genes on this clone, the previously determined genes, pht1, cdc2, his3 and act1, were found to be most highly expressed. Finally, cDNAs of all the newly identified genes were detected by RACE, proving the actual expression of these genes. The nucleotide sequence has been submitted to the EMBL database under Accession No. AB004534.
Base Sequence↗
Comprehensive cloning and expression analysis of glycolytic genes from the filamentous fungus, Aspergillus oryzae.
We cloned all the glycolytic genes from Aspergillus oryzae and analyzed their transcriptional regulation by the carbon source in the medium. The deduced amino-acid sequences of the glycolytic genes showed high identity (approximately 41-93%) to those from other lower eukaryotes. Genomic Southern hybridization indicated that all the genes existed as a single copy in the genome. Comparison of mRNA levels between mycelia grown on glucose and on pyruvate showed that most of the A. oryzae glycolytic genes were induced by glucose in the medium. The overall expression profiles of the A. oryzae glycolytic genes resembled those of Saccharomyces cerevisiae. The expression of one of the phosphofructokinase genes (pfkB), however, was repressed by glucose while both PFK1 and PFK2 were induced in S. cerevisiae. These findings indicate that further analysis of the transcriptional regulation of the A. oryzae glycolytic genes will be useful for investigating the evolutionary change of transcription regulation in lower eukaryotes and to construct promoters for industrial applications.
Aspergillus oryzae↗
Method for stretching DNA molecules on mica surface in one direction for AFM imaging.
An efficient method was developed to stretch DNA molecules on an atomically flat surface for AFM imaging. This method involves anchoring DNA molecules from their 5' ends to amino silanized mica surfaces. N-Succinimidyl6-[3'-(2-pyridyldithio) propionamido]hexanoate (LC-SPDP), a heterobifunctional cross-linker with a flexible spacer arm was used for this purpose. The immobilization process was carried out by introducing a thiol group to the 5' end of DNA by PCR. Thiolated molecules were then reacted with the cross linker to conjugate with its 2-pyridyl disulphide group via sulfhydryl exchange. The resulting complex was deposited on amino silanized mica where NHS-ester moiety of the cross linker reacted with the primary amino group on the surface. Samples were washed by a current of water and dried by an air jet in one direction parallel to the surface. DNA molecules were shown to be fully stretched in one direction on imaging them by AFM.
Aluminum Silicates↗
Affinity selection of DNA-binding proteins displayed on bacteriophage lambda.
Two transcription factors, human ATF1, its DNA-binding domain (ATF1BD), and the DNA-binding domain (GAL4BD) of the yeast GAL4 protein, were displayed on the surface of bacteriophage lambda vectors and efficiently selected by DNA fragments immobilized in microtiter wells. The DNA-binding proteins are fused to the carboxy terminus of the tail protein gpV and head protein gpD of the vectors, lambdafoo and lambdafooDc, respectively. After a single round of affinity selection, the fusion phages were successfully enriched 60- to 4,000-fold over the vector phages. Further, the GAL4BD fusion phages were enriched 5- and 15-fold by affinity selection using specific DNA as probes over nonspecific DNA when expressed on lambdafooDc and lambdafoo, respectively. The ATF1BD fusion phages were also sequence-specifically enriched greater than 4-fold when displayed on lambdafoo. These results suggest that the lambdafoo display system is useful for in vitro studying of protein-DNA interactions and may be applied to screening of DNA-binding protein from complex cDNA libraries through DNA-binding affinity.
Activating Transcription Factor 1↗
The effects of absorption enhancers on the pulmonary absorption of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in rats.
Pulmonary absorption of recombinant human granulocyte colony-stimulating factor (rhG-CSF) with various surfactants and protease inhibitors were examined in rats. The relative bioavailabilities of rhG-CSF with surfactants, such as polyoxyethylene 9-lauryl ether (Laureth-9) and sodium glycocholate (SGC), after intratracheal (i.t.) administration by intravenous (i.v.) and subcutaneous (s.c.) means were 37% (i.v.), 88% (s.c.), 84% (i.v.) and 197% (s.c.), respectively. These values were evaluated from the ratio of the area under the curve (AUC) of the plasma rhG-CSF concentration versus time for 8 h. In the presence of various kinds of protease inhibitors, such as (p-amidinophenyl) methanesulfonyl fluoride x HCl (p-APMSF), aprotinin and bestatin, an increase in the plasma rhG-CSF concentration was observed, and the effect with p-APMSF was maximal. The relative bioavailabilities of rhG-CSF with p-APMSF after i.t. administration by i.v. and s.c. means were increased about 2-fold. To clarify the absorption mechanism of rhG-CSF, rhG-CSF was intratracheally administered with both Laureth-9 and p-APMSF. The AUC of rhG-CSF increased with both agents, and was approximately equal to that with SGC, which has both an enhancing effect on membrane permeation and an inhibitory effect on enzymatic degradation after i.t. administration. Consequently, it was considered that permeation and enzymatic degradation were rate-determining steps in the pulmonary absorption of rhG-CSF after i.t. administration.
Absorption↗
Screening search for organic fluorophores: syntheses and fluorescence properties of 3-azolyl-7-diethylaminocoumarin derivatives.
In order to find a highly sensitive fluorophore, 3-azolyl-7-diethylaminocoumarin derivatives were synthesized. Both the absorption and fluorescence maxima of the coumarin-thiazole compounds showed red shifts with increases of the molar absorptivities and fluorescence intensities, in comparison with those of the corresponding coumarin-oxazole compounds. Among them, 3-(5-ethoxycarbonyl-1,3-thiazol-2-yl)-7-diethylamino-2H-chromen -2-one (3e) was one of the most promising candidates as a fluorophore accessible for analytical purposes in the fields of analytical and biological chemistry.
Azoles↗
Molecular cloning of genomic DNA for fructose-1,6-bisphosphatase. From Aspergillus oryzae.
We have cloned and sequenced an Aspergillus oryzae genomic DNA fragment that encodes a fructose-1,6-bisphosphatase gene (fbpA) with the aim of studying transcriptional regulation mechanisms involved in basic metabolism. Expression of fbpA was repressed in the presence of glucose, but not in the presence of pyruvate or sodium acetate in the medium. The CreA and FacB element found in the fbpA 5'-flanking region may be important in fbpA regulation.
Amino Acid Sequence↗
Molecular cloning and characterization of rpbA encoding RNA polymerase II largest subunit from a filamentous fungus, Aspergillus oryzae.
We have cloned rpbA encoding the RNA polymerase II largest subunit (polIIL) from a filamentous fungus, Aspergillus oryzae. The rpbA product included eight highly conserved regions and the carboxyl-terminal domain (CTD). A. oryzae polIIL CTD with 184 amino acids was composed of 25 CTD consensus repeats, which was a similar number to those of lower eukaryotes. The amino acids in each repeat of A. oryzae polIIL, however, conformed less to the CTD consensus than those of polIILs from other lower eukaryotes.
Amino Acid Sequence↗
Histopathological study of the effects of a single intratracheal instillation of surface active agents on lung in rats.
Pulmonary drug administration of most peptide/protein drugs is characterized by low bioavailability due to low permeability. Surface active agents have been tested as an absorption enhancer, but few studies have been carried out on the local toxicity of these additives. In the present study, to clarify the toxic effects of surface active agents on the lung, a relatively high concentration (1%) of polyoxyethylene 9 lauryl ether (Laureth-9) and sodium glycocholate (SGC) was given to rats in a single intratracheal instillation (100 microliters/rat), and the lung was evaluated histopathologically. In the rats treated with Laureth-9, lung lesions were observed in the bronchi to alveoli. At 1 day after administration, edema, hemorrhage and inflammatory cell infiltration due to degeneration and desquamation of epithelium were observed. At 3 and 7 days after administration, the wound healing process resulting from the lung injury, such as hyperplasia of epithelium and sporadic fibrosis, was noted. SGC also induced lung lesions with a similar histopathological nature, whereas the lesions were mostly confined to the alveoli. These results suggest that surface active agents induce acute inflammation of the lung by intratracheal instillation, that the distribution of lesions is different among surface active agents, and moreover that pathological examination is indispensable for clarifying local toxicity of absorption enhancers in pulmonary drug-delivery studies.
Absorption↗
[Age-related clinical features in ischemic colitis].
Ischemic colitis has been considered to have relatively high prevalence in the elderly population with underlying vascular disorder such as hypertension. However, this disease has been recently reported increased in the young population so that it is not necessarily limited to the aged. The aim of the present study was to elucidate the characteristics of age-related clinical features in ischemic colitis. The subjects consisted of 30 patients with ischemic colitis admitted to our hospital during the last 5 years. They were divided into the aged group more than 65 years old and the young group aged 65 or less. As a result, there were no significant differences in symptoms, resulted serological examination, endoscopic findings, and treatment period. Lesion sites were more extended in the aged group. Concerning underlying disease and etiologic factors, the vascular factor was important in the aged group, while the peristaltic factor, especially constipation was important in the young group. Ten of the 30 patients had habitual constipation, and the aged group had a high percentage of paralytic constipation, while the young group had a high rate of spastic constipation. Many patients with paralytic constipation had a history of underlying diseases and laparotomy, while the patients with spastic constipation did not have such a history. Therefore, it is presumed that the spastic type of constipation is an etiologic factor in ischemic colitis.
Adult↗
Rapid detection of homologously integrated DNA fragments and accurate quantitation of their copy number in transgenic Aspergillus oryzae by PCR.
A simple and rapid method for the analysis of artificially introduced DNA fragments has been developed using competitive PCR and long and accurate PCR. The locus and the copy number of the DNA fragments in each Aspergillus oryzae transformant could be detected more rapidly and accurately by this method than by the conventional Southern hybridization method.
Journal Article↗
Cause of idiopathic scoliosis.
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Adolescent↗
Mesoporous Magnetic Materials Based on Rare Earth Oxides.
A novel family of mesoporous oxides of rare earth metals (Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu) were synthesized with specific surface areas of 253-348 m(2) g(-1) and pore diameters of 2.5-3.0 nm (see Scheme). The mesoporous solids and their surfactant mesophase precursors are paramagnetic and exhibit a magnetic anomaly due to the mesostructured atomic arrangement.
Journal Article↗
Pathogenesis of idiopathic scoliosis. Experimental study in rats.
STUDY DESIGN: A radiographic examination of pinealectomized rats to observe the development of scoliosis and halt the condition by administration of melatonin. OBJECTIVES: To discover whether pinealectomy has the same effect in mammals as shown in the chicken, and to determine whether the bipedal condition is important for development of scoliosis. SUMMARY OF BACKGROUND DATA: Pinealectomizing chickens shortly after hatching consistently resulted in scoliosis closely resembling human idiopathic scoliosis. It has not been determined whether this phenomenon is restricted solely to chickens, or if this experimental model is applicable to other animals, especially those more closely related to humans. METHODS: A sham operation in five bipedal rats served as the control in this study. Pinealectomy was performed in 10 quadrupedal rats, pinealectomy in 20 bipedal rats, and pinealectomy with implantation of melatonin pellet in 10 bipedal rats. Spinal radiographs were used to measure the degree of scoliosis at 3 months after surgery. RESULTS: Scoliosis developed only in pinealectomized bipedal rats and not in quadrupedal rats. It developed in none of the sham operation group and in only 1 of 10 pinealectomized bipedal rats with melatonin treatment. CONCLUSIONS: Melatonin deficiency secondary to pinealectomy alone does not produce scoliosis if the quadrupedal condition is maintained. The bipedal condition, such as that in chickens or humans, plays an important role in the development of scoliosis. The findings suggest a critical influence of a postural mechanism for the development of scoliosis.
Animals↗
Cloning and characterization of a gene (arpA) from Aspergillus oryzae encoding an actin-related protein required for normal nuclear distribution and morphology of conidiophores.
We have isolated the arpA gene from Aspergillus oryzae as a homologue of the Neurospora crassa ro-4 gene. In N. crassa, mutations in the ro-4 gene, which encodes a major component of the dynactin complex Arp1, causes curling of hyphae and abnormalities in nuclear distribution. The arpA gene contains two introns and encodes a polypeptide of 381 amino acids, with a 78% sequence identity to the N. crassa Arp1. Overexpression of the arpA gene causes a defect in nuclear migration into elongating hyphae of germlings in A. oryzae. We constructed arpA disruptant strains of A. oryzae. The arpA null mutants showed poor growth and hyper-branched mycelia, as well as a nuclear distribution defect. Scanning electron microscopy revealed that the arpA null mutant has an aberrant conidiophore morphology with irregular phialides.
Actins↗