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Biomedical subjects

K Fukui

Publications and source records attributed to K Fukui.

At least 73 records · Page 4Linked to original sources

[Surgical treatment of infectious endocarditis complicated by discrete subaortic stenosis: report of a case].

A 39-year-old male with a history of cardiac murmur from early childhood was diagnosed as having infectious endocarditis (IE) complicated by discrete subaortic stenosis (DSS). Echocardiography revealed severe aortic regurgitation, subaortic membranous structure and mild mitral regurgitation. The pressure gradient across the subaortic stenosis was 105 mmHg according to continuous Doppler wave ultrasonography. Aortic valve replacement and resection of subaortic membranous tissue and mitral annuloplasty were performed. Postoperative cardiac catheterization demonstrated that the hemodynamic data were remarkably improved, and the patient was free of symptoms. He is currently well at 6 months after the operation.

Adult↗

[Multiple focal cortical dysplasias presenting with intractable epilepsy: case report].

A 14-year-old female, who had intractable epilepsy associated with multiple focal cortical dysplasias (FCD), was reported. She developed intractable epilepsy at the age of 7 and was diagnosed as having frontal lobe epilepsy based on the seizure semiology and interictal EEG. MRI revealed multiple lesions in the right frontal, bilateral occipital and left parietal lobes. EEG demonstrated that ictal discharge was preceded by spike on the right frontal region and FDG-PET showed hypometabolic area in the right frontal lobe. Chronic subdural electrode recordings from the right frontal lobe indicated that ictal onset zone was located around the right frontal lesion, especially frontal tip and base, and these areas including the lesion were resected. Postoperatively, residual seizure was noted although seizure frequency was decreased. It is well known that, postoperatively, satisfactory seizure outcome can be obtained in patients with FCD. However, further investigation in terms of surgical indication and strategies for multiple FCD should be needed.

Adolescent↗

Chromosome painting as a tool for rice genetics and breeding.

Chromosome painting and genomic in situ hybridization (GISH) are both effective methods for basic genetic research and practical breeding. These methods were applied even in the typically small chromosomes of rice. This manuscript describes in detail, highly reproducible, complete protocols for chromosome painting and GISH in rice chromosomes. Examples of useful applications of these methods are also presented.

Breeding↗

Both corticotropin releasing factor and neuropeptide Y are involved in the effect of orexin (hypocretin) on the food intake in rats.

Orexin (hypocretin) is a peptide that has been found to stimulate food intake in rats. However, we have recently demonstrated that orexin stimulates the release of corticotropin releasing hormone (CRF) which has been known to decrease the food intake. Therefore, we examined the mechanism of effect of orexin on food intake. Although the other appetite stimulating peptides; neuropeptide Y (NPY), agouti-related peptide (AGRP) and one of the growth hormone releasing secretagogue (GHRP-6) stimulated dose-dependently the food intake during 2 h in the early light period, orexin did not increase significantly the food intake. No significant increase was also observed during 2 h in the early dark period. However, pretreatment with alpha-helical CRF, an antagonist of CRF, or anti-CRF antiserum resulted in significant increase of food intake by orexin. Orexin-stimulated feeding under these conditions was blocked by NPY Y1 receptor antagonist (1229U91). In an 8 h-fasting rat, anti-orexin serum decreased slightly the food intake. These results suggest that effect of orexin on the food intake may be complex because of orexin-CRF and orexin-NPY linkage.

Agouti-Related Protein↗

Brain regions involved in verbal or non-verbal aspects of facial emotion recognition.

To depict the neural substrates for facial emotion recognition and to determine whether their activation is confounded by a verbal factor, we studied eight normal volunteers with functional magnetic resonance imaging (fMRI). Verbal and non-verbal sample stimuli were used in a facial emotion matching task and a gender matching task (control condition). Compared with the gender tasks, the emotion tasks significantly activated the right ventral prefrontal cortex, the right lingual cortex, and the left lateral fusiform cortex, irrespective of sample stimuli. The visual association cortices showed a significant interaction between the task and the material presented, as the activation for verbal materials was higher than for non-verbal materials during the emotion matching tasks. By contrast, no significant interaction was found in the right ventral prefrontal cortex. These results suggest that the verbal factor has a different effect on the neural networks for facial emotion, processing.

Brain Mapping↗

Pseudogap formation in the intermetallic compounds (Fe1-xVx)3Al

Optical conductivity data of the intermetallic compounds (Fe1-xVx)3Al ( 0</=x</=0.33) reveal that their density of states around the Fermi energy ( E(F)) is strongly reduced as x is increased. In particular, Fe2VAl ( x = 0.33) has a deep, well-developed pseudogap of 0.1-0.2 eV at E(F) and a small density ( approximately 5x10(20) cm(-3)) of carriers, which is highly unusual for intermetallic compounds. It is shown that the pseudogap results from the band structure of Fe2VAl, rather than from temperature-dependent correlation effects. Based on the present results, we propose a simple model that consistently explains both the semiconductorlike transport and the metallic photoemission results previously observed for Fe2VAl.

Journal Article↗

NBP1 (Nap1 binding protein 1), an essential gene for G2/M transition of Saccharomyces cerevisiae, encodes a protein of distinct sub-nuclear localization.

Nap1p is identified in mammalian cell extract by its intrinsic activity to facilitate nucleosome assembly in vitro in the physiological ionic condition. The homologous proteins are present in most eukaryotes, and their functional analyses in vitro have suggested that they are necessary to keep proper nucleosome structures in transcription and replication. This protein is also identified for its interaction with Clb2p in vitro. To address the function of Nap1p in vivo, we have surveyed for proteins to interact with Nap1p by two-hybrid system and obtained two genes, NBP1 and NBP2 (Nap1 Binding Protein 1 and 2). NBP1 is an essential gene and encodes a novel protein consisting of 319 amino acids, with a coiled-coil structure in the center of the predicted amino acid sequence. Several A-kinase dependent phosphorylation sites and Cdc28p kinase-dependent sites are also observed. By isolating the temperature-sensitive mutant, we demonstrate that the nuclear division at a non-permissive temperature is delayed and that the population of cells with a large bud carrying a single nucleus with a short spindle are increased. This mutant also confers resistance against benomyl, a microtubule-destabilizing agent. Judging from the green fluorescent protein (GFP) signal fused with Nbp1p, this protein localizes in the nucleus as one or two tiny dots.

Amino Acid Sequence↗

Design and properties of human D-amino acid oxidase with covalently attached flavin.

An "artificial flavinylation" approach was developed to replace a native noncovalent flavin prosthetic group with a covalently attached flavin analogue in recombinant human d-amino acid oxidase. The protein residue Gly-281 was replaced with Cys by site-directed mutagenesis, followed by reaction between mutated apoenzyme and the thiol-reactive flavin analogue, 8-methylsulfonyl FAD. The stoichiometric process of flavin attachment was accompanied by gain in enzymatic activity, reaching up to 26% activity of the recombinant native enzyme. The steady-state kinetic data together with the results of limited proteolysis and benzoate-binding studies suggest that, although mutation perturbs protein structural and catalytic properties, the flavinylation alone does not have any negative impact. We conclude that, despite the implemented restraints on its mobility, the covalently attached flavin is properly positioned within the protein active site and acts efficiently during d-amino acid oxidase catalytic turnover.

Base Sequence↗

Nafamostat mesilate modulates the release of platelet-activating factor during left ventricular assistance with hemofiltration in canine heart failure.

OBJECTIVE: The enhanced generation of various chemical mediators is regarded as one of the mechanisms by which severe heart failure progresses to multiple organ failure. Platelet-activating factor is a phospholipid mediator which plays an important role in inflammatory reactions and circulatory shock. We studied the changes in platelet-activating factor levels in a canine heart failure model treated with a left ventricular assist device and hemofiltration, and assessed the effect of a protease inhibitor, nafamostat mesilate. METHODS: Twenty dogs underwent multiple coronary ligations, and at 2 hours after the ligations they were maintained on left ventricular assist device support with continuous hemofiltration. The animals were divided into two groups: a nafamostat group (n = 10) that received nafamostat mesilate (2 mg/kg/hr), and a control group (n = 10) that received vehicle only. RESULTS: The blood platelet-activating factor level, before coronary ligations, in the control and nafamostat groups was 2.3 +/- 0.4 and 2.0 +/- 0.7 ng/ml, respectively, and the coronary ligations had little effect on the platelet-activating factor. However, after the initiation of left ventricular assist device, the platelet-activating factor in the control group (5.6 +/- 2.2) was significantly higher (p < 0.05) than that in the nafamostat group (1.1 +/- 0.3). Nafamostat administration was also effective in controlling the increase in the blood lactate level. Hemofiltration did not change the platelet-activating factor. CONCLUSIONS: We concluded that platelet-activating factor may play a critical role in the development of severe heart failure with left ventricular assistance, and nafamostat administration is likely to be beneficial in such a critical condition by suppressing the platelet-activating factor level.

Animals↗

Morphogenesis in cucumber seedlings is negatively controlled by gravity.

Seedlings of most cucurbitaceous plants develop a peg (protuberance caused by cell outgrowth) on the transition zone between the hypocotyl and root. The peg is necessary for removing the seed coat after germination. In our spaceflight experiments on the STS-95 space shuttle, Discovery, we found that cucumber (Cucumis sativus L.) seedlings grown under microgravity conditions developed two pegs symmetrically at the transition zone. Thus, cucumber seedlings potentially develop two pegs and do not require gravity for peg formation itself, but on the ground the development of one peg is suppressed in response to gravity. This may be considered as negative control of morphogenesis by gravity.

Cucumis sativus↗

Quantification of total genomic DNA and selected repetitive sequences reveals concurrent changes in different DNA families in indica and japonica rice.

This paper describes a fluorescence in situ hybridization (FISH) analysis of three different repetitive sequence families, which were mapped to mitotic metaphase chromosomes and extended DNA fibers (EDFs) of the two subspecies of rice (OrYza sativa), indica and japonica (2n = 2x = 24). The repeat families studied were (1) the tandem repeat sequence A (TrsA), a functionally non-significant repeat; (2) the [TTTA-GGG]n telomere sequence, a non-transcribed, tandemly repeated but functionally significant repeat; and (3) the 5S ribosomal RNA (5S rDNA). FISH of the TrsA repeat to metaphase chromosomes of indica and japonica cultivars revealed clear signals at the distal ends of twelve and four chromosomes, respectively. As shown in a previous report, the 17S ribosomal RNA genes (17S rDNA) are located at the nucleolus organizers (NORs) on chromosomes 9 and 10 of the indica cultivar. However, the japonica rice lacked the rDNA signals on chromosome 10. The size of the 5S rDNA repeat block, which was mapped on the chromosome 11 of both cultivars, was 1.22 times larger in the indica than in the japonica genome. The telomeric repeat arrays at the distal ends of all chromosome arms were on average three times longer in the indica genome than in the japonica genome. Flow cytometric measurements revealed that the nuclear DNA content of indica rice is 9.7% higher than that of japonica rice. Our data suggest that different repetitive sequence families contribute significantly to the variation in genome size between indica and japonica rice, though to different extents. The increase or decrease in the copy number of several repetitive sequences examined here may indicate the existence of a directed change in genome size in rice. Possible reasons for this phenomenon of concurrent evolution of various repeat families are discussed.

Blotting, Southern↗

Ribosomal DNAs: an exception to the conservation of gene order in rice genomes.

rDNA (18S-5.8S-25S rDNA) and 5S rDNA loci were visualized on the chromosomes of six species of the genus Oryza by fluorescence in situ hybridization (FISH) and the labeled rice chromosomes were identified based on their condensation patterns. As a result, the chromosomes harboring rDNA and/or 5S rDNA loci were determined in the complement for all the known rice genomes. Variation in the location of the rDNA loci indicated the transpositional nature of the rDNAs in the genus Oryza, as also suggested in Triticeae and Allium. Comparative analysis of the locations of rDNA loci among rice, maize and wheat revealed that variability in the physical location of the rDNA loci was characteristic of the genus Oryza and also of the genera of Gramineae. This variability in the location of the rDNA loci between evolutionarily related species is in sharp contrast to the conservation of the general order of genes in their genomes.

Chromosome Mapping↗

Soy protein isolate and its hydrolysate reduce body fat of dietary obese rats and genetically obese mice (yellow KK)

This study examined in dietary obese and genetically obese rodents the effects of soy protein isolate (SPI) and its hydrolysate (SPI-H) on the rate of body-fat disappearance. Male Sprague-Dawley rats (4-16 wk old) and yellow KK mice (6-10 wk old) were made obese by feeding high-fat diets containing 30% fat. They were then fed energy-restricted, low-fat (5.0%), and high-protein (35% casein, SPI, or SPI-H) diets for 4 wk at 60% of the level of the energy intake of rodents on laboratory chow. The body-fat contents of rats and mice fed a high-fat diet were 27.3 and 33.6 g/100 g body weight, respectively, at the end of the obese period. For rats, the apparent absorbability of dietary energy and fat was significantly lower in the SPI and SPI-H groups than in the casein group, but vice-versa for nitrogen balance. Body-fat content in mice fed SPI and SPI-H diets was significantly lower than in those fed the casein diet. In rats, plasma total cholesterol level was lower with the SPI-H diet, and plasma glucose level was lower with the SPI and SPI-H diets than with the casein diet. These results indicate that SPI and SPI-H are suitable protein sources in energy-restricted diets for the treatment of obesity.

Adipose Tissue↗

Effect of polyamines on histone-induced DNA compaction of hyperthermophilic archaea.

The effect of polyamines on histone-mediated DNA compaction was examined in vitro with archaeal histone HpkA from Pyrococcus kodakaraensis KOD1. An agarose gel mobility-shift experiment indicated that histone-bound DNA (compacted DNA) was further compacted by addition of a polyamine (putrescine, spermidine, or spermine) or its acetylated form (N-acetylputrescine, N1-acetylspermidine, N8-acetylspermidine, or N1-acetylspermine) when the mixture was incubated at above 75 degrees C. Spermine was most effective in compaction enhancement among all the polyamines tested. A high concentration of potassium ion (1.0 M) did not stabilize the compacted form of DNA even though double-stranded DNA was stably maintained against thermal denaturation at elevated temperatures under this condition. It appears likely that multivalent polyamines have a nucleosome maintenance function in hyperthermophilic archaea in high-temperature environments.

Journal Article↗

Alteration of product specificity of cyclodextrin glucanotransferase from Thermococcus sp. B1001 by site-directed mutagenesis.

Cyclodextrin glucanotransferase (CGTase) from the hyperthermophilic archaeon Thermococcus sp. B1001 catalyzed the production predominantly of alpha-cyclodextrin (CD) from starch (Tachibana, Y. et al., Appl. Environ. Microbiol., 65, 1991-1997, 1999). The CGTase gene (cgtA) from this strain was cloned and sequenced. It was composed of 2217 nucleotides, and encoded a protein (739 amino acids) with a molecular mass of 83,240 Da. Recombinant CgtA expressed in Escherichia coli also catalyzed the production predominantly of alpha-CD from starch, as did native CgtA from strain B1001. Based on a substrate binding model of Bacillus circulans no. 8 CGTase, Tyr100, Trp191 and Tyr267 were specified to locate the spiral amylose and to minimize the size of the CD by saccharide aromatics interaction. In order to determine the critical residue for catalyzing production predominantly of alpha-CD, site-directed mutations were introduced in CgtA (Y100W, Tyr100-->Trp; W191Y, Trp191-->Tyr; W191F, Trp191-->Phe; Y267W, Tyr267-->Trp; Y267F, Tyr267-->Phe). Analysis of the reaction products by HPLC revealed that the mutant enzyme Y267W produced more beta- and gamma-CD than the wild-type enzyme. However, the other mutants still produced high levels of alpha-CD, suggesting that Tyr267 plays a critical role in alpha-CD production catalyzed by B1001 CGTase.

Journal Article↗

Site-specific accumulation of a LINE-like retrotransposon in a sex chromosome of the dioecious plant Cannabis sativa.

Male-associated DNA sequences were analysed in hemp (Cannabis sativa L.), a dioecious plant with heteromorphic sex chromosomes. A male-associated DNA sequence in C. sativa (MADC1) and its flanking sequence encoded a reverse transcriptase that was strongly homologous to those of LINE-like retrotransposons from various plants and other organisms, as well as another open reading frame (ORF). Fluorescence in situ hybridization (FISH) with MADC1 as probe, which yielded strong signals specific for male genomic DNA in gel blot analysis, generated a clear doublet signal at the end of the long arm of the Y chromosome. FISH using pachytene chromosomes of pollen mother cells at meiotic prophase I revealed that pairing of X and Y chromosomes occurred at the short arm of the Y chromosome where MADC1 was not present. Furthermore, FISH using extended DNA fibers, with MADC1 and its flanking DNA as probes, revealed that 100 to 200 copies of the retrotransposon were located in tandem on the Y chromosome. These results support the hypothesis that accumulation of a specific LINE-like retrotransposon at the terminal region of the long arm of the Y chromosome might be one cause of heteromorphism of sex chromosomes.

Amino Acid Sequence↗