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Biomedical subjects

H Fukuda

Publications and source records attributed to H Fukuda.

At least 145 records · Page 8Linked to original sources
PubMed2000

Preparation of Thermus thermophilus holo-chaperonin-immobilized microspheres with high ability to facilitate protein refolding.

Carboxylated poly(styrene/acrylamide) (P(St/AAm)-H) microspheres with different acrylamide contents were prepared by emulsifier-free emulsion polymerization. Thermus thermophilus holo-chaperonin (cpn) was covalently immobilized onto these microspheres with high yield. The T. thermophilus holo-cpn-immobilized microspheres were used for refolding of guanidine hydrochloride (Gdn-HCl)-denatured enzymes and showed sufficiently high ability to facilitate refolding of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase (G6PD) and pig heart lactate dehydrogenase (LDH) at 30 degrees C and Bacillus stearothermophilus LDH at 60 degrees C. The specific ability of T. thermophilus holo-cpn-immobilized microspheres increased with increasing immobilization amount and reached plateau at around 10-15 mg/m(2). When the immobilization amounts of T. thermophilus holo-cpn were approximately 10 mg/m(2), the microspheres retained sufficiently high ability to facilitate protein refolding during repeated use. Therefore, the P(St/AAm)-H microspheres on which approximately 10 mg/m(2) of T. thermophilus holo-cpn is immobilized are very effective for refolding of various (Gdn-HCl)-denatured enzymes over a wide temperature range.

Acrylic Resins↗
PubMed2000

Oxidative stress-related proteins A170 and heme oxygenase-1 are differently induced in the rat cerebellum under kainate-mediated excitotoxicity.

A170 and heme oxygenase-1 (HO-1) are characterized as oxidative stress-inducible proteins whose induction depends upon common transcription factor via antioxidant responsive element. We investigated the expression of A170 and HO-1 in the cerebellum after kainate administration. In situ hybridization showed constitutive expression of A170 and HO-1 mRNA in Purkinje cell layer; mild induction of A170 or HO-1 was detected, respectively, 8 or 24 h after kainate administration. Immunohistochemical studies also demonstrated that constitutive expression and the induction of A170 protein in Purkinje cells; the induction of HO-1 protein was detected in Bergmann glia but not in Purkinje cells. Thus, the transcription factors involved in the induction of A170 might be different from those in the induction of HO-1 under kainate-mediated excitotoxicity. The existence of cell type-specific stress response was suggested.

Animals↗
PubMed2000

Isolation of a vascular cell wall-specific monoclonal antibody recognizing a cell polarity by using a phage display subtraction method.

Using a strategy consisting of (i) the isolation of cell walls from synchronously differentiating cells of Zinnia, (ii) the generation of mAbs with an antibody phage display method, and (iii) screening with a subtraction method, we isolated mAbs recognizing vascular development-specific cell wall components without prior antigen identification. One of the isolated mAbs, designated CN 8, recognized a cell wall component contained in the hemicellulosic fraction. Immunohistochemical analyses showed that the CN 8 epitope was localized to the cell wall of immature tracheary elements and xylem parenchyma cells. In immature tracheary elements, the CN 8 epitope had a polarized localization pattern regardless of whether the cells are formed as parts of vessels in situ or as single tracheary elements in vitro, suggesting that cell polarity autonomously formed on the cell wall may function in tracheary element differentiation.

Amino Acid Sequence↗
PubMed2000

Retention of words in long-term memory: a functional neuroanatomical study with PET.

We used PET to identify brain regions associated with retention of verbal materials in long-term memory. During a PET scan, subjects repeated many sets of words one after another. In a retention condition, they were simultaneously required to retain 10 key words that were irrelevant to the repetition task. Significant increases in regional cerebral blood flow during the retention condition were found in bilateral parahippocampal regions, the left prefrontal and parietal association cortices, the supplementary motor area, the neostriatum and the cerebellum. We clearly demonstrated that retention of verbal materials was accompanied by neural activities in the medial temporal lobes. We also showed that, in the early phase, retention of words in long-term memory recruited left cortical areas surrounding those relevant to verbal short-term memory.

Acoustic Stimulation↗
PubMed2000

Passive and active recognition of one's own face.

Facial identity recognition has been studied mainly with explicit discrimination requirement and faces of social figures in previous human brain imaging studies. We performed a PET activation study with normal volunteers in facial identity recognition tasks using the subject's own face as visual stimulus. Three tasks were designed so that the activation of the visual representation of the face and the effect of sustained attention to the representation could be separately examined: a control-face recognition task (C), a passive own-face recognition task (no explicit discrimination was required) (P), and an active own-face recognition task (explicit discrimination was required) (A). Increased skin conductance responses during recognition of own face were seen in both task P and task A, suggesting the occurrence of psychophysiological changes during recognition of one's own face. The left fusiform gyrus, the right supramarginal gyrus, the left putamen, and the right hypothalamus were activated in tasks P and A compared with task C. The left fusiform gyrus and the right supramarginal gyrus are considered to be involved in the representation of one's own face. The activation in the right supramarginal gyrus may be associated with the representation of one's own face as a part of one's own body. The prefrontal cortices, the right anterior cingulate, the right presupplementary motor area, and the left insula were specifically activated during task A compared with tasks C and P, indicating that these regions may be involved in the sustained attention to the representation of one's own face.

Adult↗
PubMed2000

A PET study of visuomotor learning under optical rotation.

We measured the regional cerebral blood flow (rCBF) in six healthy volunteers with PET (positron emission tomography) and H(15)(2)O to identify the areas of the human brain involved in sensorimotor learning. The learning task was visually guided reaching with sensorimotor discrepancy caused by optical rotation. PET measurements were performed in the early and late stages of the adaptation to the sensorimotor perturbation. Control measurements were obtained during an eye movement task and a reaching task without optical rotation. The rCBF data of each learning stage were compared to those of both control conditions. During the early stage, rCBF increases were detected in the rostral premotor cortex bilaterally, the posterior part of the left superior parietal lobule (SPL), and the right SPL including the intraparietal sulcus (IPS). During the late stage, rCBF increases were detected in the left caudal premotor area, the left supplementary motor area proper, the left SPL, the right SPL including the IPS, and the right postcentral sulcus extending to the inferior parietal lobule. These results reveal that sensorimotor learning accompanies changes in the recruited cortical areas during different stages of the adaptation, reflecting the different functional roles of each area for different components of adaptation, from learning of new sensorimotor coordination to retention or retrieval of acquired coordination.

Adult↗
PubMed2000

Correlation between human personality and neural activity in cerebral cortex.

Personality traits are a variance of behavioral patterns among individuals and may reflect a variance of brain activity, but their neurobiological explanation is still a matter of debate. Cloninger proposed three dimensions of personality traits, each of which has strong correlation with activity in a specific central monoaminergic system. Although this theory has been supported by physiological and genetic studies, it is still unclear how these personality parameters are correlated with the activity of the cortical networks which control human behavior. Here we measured the regional cerebral blood flow (rCBF) at rest in 30 normal volunteers who completed the personality inventory of Cloninger. Voxel-by-voxel analysis was employed to identify cortical regions where the rCBF showed significant correlation with any of the three personality parameters. Statistically significant correlation was observed in several paralimbic and neocortical regions and was consistent with the assumed monoaminergic influence on neural activity and the distribution of its projections, in each personality dimension. The results suggest that activity in a variety of cortical regions is associated with human personality traits and lend support to Cloninger's theory concerning central monoaminergic influence on human personality traits.

Adult↗
PubMed2000

The effect of verbal feedback on motor learning--a PET study. Positron emission tomography.

The purpose of this study was to investigate brain mechanisms underlying feedback effects on motor learning. We measured human brain activity using positron emission tomography (PET) during length-of-line drawing tasks in the presence or absence of verbal feedback, i.e., information on the precision of motor performance. The average error in responses was significantly lower and the percentage of correct responses was significantly higher in the case of tasks with feedback than those in the absence of feedback. The contralateral sensorimotor, premotor, supplementary motor, the right prefrontal, bilateral parietal and temporal, and anterior cingulate cortices, and the left basal ganglia were activated during all the line-drawing tasks. The right lateral prefrontal and occipital cortices and the left basal ganglia exhibited marked increase in activity after learning. The right inferior parietal and the anterior cingulate cortices were activated in the presence of feedback which provided information on how the subjects should correct their performances. The results indicate that these brain areas may play an important role in representing knowledge of results during motor learning and that appropriate feedback may facilitate motor learning.

Adult↗
PubMed2000

Benign schwannoma of the esophagus removed by enucleation: report of a case.

A submucosal tumor of the esophagus was enucleated by a thoracotomy. A microscopic examination showed the tumor to be composed of spindle-shaped cells showing mild nuclear atypia with vague nuclear palisading and scarce mitotic figures. The tumor was surrounded by peripheral lymphoid cuffs. An immunohistochemical study demonstrated diffuse positive staining for S-100 protein in the tumor cells. The lesion was diagnosed to be an esophageal schwannoma based on these pathological features. Benign schwannoma of the esophagus has been described in five cases in four reports in the English literature. This is the sixth case diagnosed by immunohistochemical studies.

Esophageal Neoplasms↗
PubMed2000

The rates of deceleration of nuclear and organellar DNA syntheses differ in the progenitor cells of the apical meristems during carrot somatic embryogenesis.

The synthesis of DNA in nuclei and organellar nucleoids at the various stages of somatic embryogenesis in carrot (Daucus carota L. cv. Kurodagosun) was analyzed using anti-5-bromo-2'-deoxyuridine (BrdU) immunofluorescence microscopy. The active syntheses of both nuclear and organellar DNA started in the cells forming the embryo proper 3 d after the initiation of embryogenesis, but not in cells forming suspensor-like cell aggregates. In the early globular embryo, active DNA syntheses were continuously observed in the whole embryo proper, except for the progenitor cells of the root apical meristem (RAM) and shoot apical meristem (SAM). These were recognized as slowly cycling cells with a non-BrdU-labelled nucleus and strongly BrdU-labelled organellar nucleoids. At the heart- and torpedo-shaped embryo stages, both nuclear and organellar DNA syntheses were inactive in the presumptive RAM and SAM. Thus, slowing down of organellar DNA synthesis is not coupled with, but is later than, that of nuclear DNA synthesis in the progenitor cells of the embryonic RAM and SAM. These findings clearly indicate that the timing of DNA synthesis is similar in the progenitor cells of both the RAM and SAM in the early stages of somatic embryogenesis.

Cell Nucleus↗
PubMed2000

Preparation of high activity whole cell biocatalyst by permeabilization of recombinant flocculent yeast with alcohol.

Flocculent yeast Saccharomyces cerevisiae YF234 (MATa ura3-52 trp1Delta2 his ade 2-1 can1-100 sta1 FLO8) cells overexpressing glyoxalase I and having strong flocculation ability were permeabilized with isopropyl alcohol and ethanol under various conditions. The treatment with 40% isopropyl alcohol significantly improves the initial reaction rates of recombinant flocculent yeast cells. Moreover, the reactivity of permeabilized flocculent yeast cells was similar to that of dispersed cells with EDTA. On the other hand, the flocculation ability of yeast cells was not affected by the treatment with alcohol solutions of various concentrations and treatment time length. Therefore, the recombinant flocculent yeast cells permeabilized with alcohol are very effective whole cell biocatalysts.

Journal Article↗
PubMed2000

Relationship between copper, zinc and metallothionein in hepatocellular carcinoma and its surrounding liver parenchyma.

BACKGROUND/AIM: Accumulation of copper (Cu) in hepatocellular carcinoma (HCC), especially in small tumors, is greater than that in the surrounding liver parenchyma. Metallothionein (MT) is considered to be present as Cu-MT, Zn,Cu-MT or Zn-MT. The aim of this study was to determine the presence and localization of Cu-MT and Zn-MT in HCC and surrounding liver parenchyma. METHODS: In 16 HCC patients, surgically resected specimens including HCC and surrounding liver parenchyma were evaluated. RESULTS: The level of Cu present in small HCC (<4 cm in diameter) was significantly greater than that in the surrounding liver parenchyma (p<0.05). However, the level of Cu in large HCC (>4 cm in diameter) was similar to that in the surrounding liver parenchyma. Analysis by Sephadex G-75 gel filtration revealed that the peak fraction due to Cu was identical to that due to MT in 14 (87.5%) of 16 HCC, the peak fraction due to Cu and Zn was identical to that due to MT in 2 (12.5%) HCC, and the peak fraction due to Zn was identical to that of MT in none of 16 HCC. CONCLUSIONS: Accumulation of Cu in small HCC, in which Cu was present as Cu-MT or Zn, Cu-MT, was greater than that in the surrounding liver parenchyma. Cu accumulation and the presence of MT in the liver may be related to carcinogenesis of HCC, because of the similarity of these findings in the experimental data of Long-Evans rats with a cinnamon-like coat color who develop HCC spontaneously.

Adult↗
PubMed2000

Clinical evaluation of sonoelasticity measurement in liver using ultrasonic imaging of internal forced low-frequency vibration.

The aim of this study is to evaluate liver elasticity noninvasively. We have already proposed an ultrasonic imaging system that can reconstruct vibration maps inside tissue under forced mechanical vibration. With this system, shear elastic properties of soft tissue can be evaluated as vibration velocities. Theoretically, these velocities increase with the increase of tissue elasticity. Sonoelasticity imaging was performed on 236 patients with chronic hepatitis and liver cirrhosis, and 50 healthy volunteers. The average of the velocities was 598.8 +/- 151.7 cm/s in healthy volunteers, 984.4 +/- 362.5 cm/s in chronic hepatitis and 1189.0 +/- 411.7 cm/s in liver cirrhosis. The average velocity of Child C group was statistically faster than those of Child A and B groups. Fibrotic rate from biopsy specimens statistically had the strongest positive correlation with velocities. With our system, the degree of liver fibrosis and function can be estimated objectively and noninvasively.

Adult↗
PubMed2000

Preparation of high-activity whole cell biocatalysts by permeabilization of recombinant yeasts with alcohol.

Recombinant yeast cells intracellularly overexpressing three different enzymes were permeabilized with alcohol under various conditions. The effects of enzyme stability in alcohol and enzyme molecular weight on the activities of permeabilized cells and enzyme leakage during incubation were examined. Saccharomyces cerevisiae YPH250 overexpressing glyoxalase I (GloI), S. cerevisiae MT8-1 overexpressing isocitrate lyase (ICL), and Pichia pastoris GS115 overexpressing beta-galactosidase (beta-gal) were used as model recombinant yeast systems. In all cases, the percentage of alcohol used for the treatment significantly affected the activity of permeabilized whole cell biocatalysts; cells showed high activity when treated with 40% isopropyl alcohol. The activity of whole cell biocatalysts was also significantly affected by the stability of the enzyme in alcohol solution; permeabilized yeast cells overexpressing ICL, which had low stability, showed rather low activity. Although the enzyme leakage from permeabilized cells was rather low in all cases, the molecular weight of the enzyme appeared to affect the extent of enzyme leakage during incubation. Permeabilized cells of P. pastoris overexpressing beta-gal (540 kDa) retained particularly high activity during incubation and could be used as an immobilized whole cell biocatalysts.

Journal Article↗
PubMed2000

Production of recombinant protein by baculovirus-infected insect cells in immobilized culture using porous biomass support particles.

Immobilization of insect cells using porous biomass support particles (BSPs) and production of a recombinant protein by the immobilized cells after infection with a baculovirus were investigated in a shake-flask culture. Sf9 cells were passively immobilized in reticulated polyvinyl formal (PVF) resin BSPs (2 x 2 x 2 mm cubes) with matrices of 60 mum mean pore diameter in situ in shake-flasks. The cell density in the BSPs was over 5 x 10(7) cells/cm3-BSP in cultures with regular replacement of the culture medium, as estimated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. After infection with a recombinant baculovirus carrying the beta-galactosidase gene, immobilized cells within the BSPs showed a high specific productivity, comparable to the maximum productivity in shake-flask cultures of non-immobilized cells, as long as nutrients in the medium were not depleted. Even when immobilized cells at a high density of 5 x 10(7) cells/cm3-BSP were infected with the baculovirus, efficient beta-galactosidase production with a high specific productivity was possible by replacing the medium at appropriate intervals to avoid nutrient depletion.

Journal Article↗
PubMed2000

A solid-state culture system using a cellulose carrier containing defined medium as a useful tool for investigating characteristics of koji culture.

The purpose of this article is to establish a new solid-state culture system that uses a defined medium and allows control of the chemical factors in the medium in solid-state cultivation. To achieve this purpose, we produced a solid substrate, in which the defined medium was packed in a cellulose carrier with agar, and named it "cellulose agar cube (CA-cube)". In order to obtain growth conditions similar to koji culture, we then determined the moisture content of the CA-cube. Five runs of column reactor operations were carried out at various moisture contents of the CA-cube in the range of 50-90%. A very similar mycelial morphology to that of koji culture was observed on the surface of CA-cube with moisture content lower than 63%. The glucoamylase production and the production ratio of glucoamylase to alpha-amylase (G/A ratio) were both increased exponentially with decreasing moisture content and reached 237.5 units per g dry solid and 13.59, respectively, at the moisture content of 50%, which are almost comparable to those of koji culture. In addition, the glucoamylase "glaB", which is mainly produced in koji culture, was observed in the CA-cube culture with low moisture content. These results indicate that growth conditions similar to koji culture could be attained by the column reactor culture with the CA-cube.

Journal Article↗