Biomedical subjects

H Fukuda

Publications and source records attributed to H Fukuda.

At least 73 records · Page 4Linked to original sources

PubMed2001

Cell type-specific roles of histone deacetylase in TR ligand-independent transcriptional repression.

Recent evidence indicates that corepressor protein with histone deacetylase (HDAC) activity mediates thyroid hormone receptor (TR) transcriptional repression. In order to examine the physiological relevance of HDAC in ligand-independent TR-mediated repression, we studied the effect of trichostatin A (TSA), a specific HDAC inhibitor, in transient transfection studies with natural reporters, and assessed the expression of TR-regulated endogenous genes. Luciferase-coupled DR4-, F2-, PAL- or GH-TREs and TRbeta1 expression vectors were cotransfected in CV-1 and GH(3) cells. We did not observe any effect of TSA on TR-induced basal repression in CV-1 cells. Instead, TSA was able to induce an increase in transcription without T(3) on all TREs tested in GH(3) cells. This increase was >7-fold on F2-, >4-fold on DR4-, and 3-fold on GH-TREs. The cotransfection of a TRbeta1 mutant that exhibits decreased affinity with N-CoR (AHT) reduced the TSA effect in GH(3) cells, demonstrating a primary role for TR/N-CoR/Sin3/HDAC complex. Next, we examined the effects of TSA on endogenous GH mRNA production in GH(3) cells by Northern blot analysis. We observed an increase of 50-70% of GH mRNA in cells treated with TSA in hypothyroid medium, and an increase of GH mRNA in T(3)-treated cells after TSA treatment. Our results show that TSA can increase the expression of endogenous genes that are susceptible to TR regulation. These results support an active role of HDAC in transcriptional repression by ligand-independent TR.

Animals↗

PubMed2001

Transcriptional regulation of insulin receptor gene promoter in rat hepatocytes.

To investigate the DNA regulatory sequences -618 to -593 (initiator ATG is +1) of the insulin receptor (IR) promoter required for IR gene expression, primary cultured hepatocytes of rats were transfected with plasmids containing the 5'-flanking sequences of the rat IR gene fused to the luciferase gene. When three copies of the nucleotides -618 to -593 of the IR promoter were transfected, the reporter activity was significantly increased in the presence of glucose and more increased in the presence of glucose/insulin. The glucose/insulin stimulation was inhibited by the addition of polyunsaturated fatty acids. These results were similar to those found earlier for the transcriptions of the fatty acid synthase, FAS(-57/-35), ATP citrate-lyase, ACL(-64/-41), and leptin(-101/-83) genes, which promoter sequences have a high similarity to IR(-618/-593). However, similarly to the leptin gene, the IR gene was more responsive to glucose stimulation than the FAS and ACL genes. Mutation of either one of the Sp1 binding sites (-618/-593) did not significantly affect the transcription, whereas mutation of three or four Sp1 binding sites resulted in a loss of responsiveness to glucose/insulin. Gel mobility shift assays revealed that nuclear factor(s) from rat liver specifically formed complexes with the sequence of IR(-618/-593). By antibody supershift assays, the transcription factors Sp1and Sp3 were found to bind with the IR(-618/-593). The IR, leptin, ACL and FAS genes contain common DNA-sequences responsible for the glucose/insulin-stimulation, suggesting that these genes are similarly regulated.

Animals↗

PubMed2001

Identification of novel membrane-bound phospholipase D from Streptoverticillium cinnamoneum, possessing only hydrolytic activity.

A membrane-bound phospholipase D (PLD) has been identified and isolated in a soluble form from an actinomycete, Streptoverticillium cinnamoneum. The enzyme has a monomeric structure with a molecular size of about 37 kDa, being the smallest among the enzymes so far reported. The enzyme catalyzes the hydrolysis of phosphatidylethanolamine and phosphatidylserine as preferred substrates, but not the transphosphatidylation reaction of their phospholipid groups to ethanol. Together with the absence of immunochemical cross-reactivity, these enzymatic properties demonstrate that the membrane-bound enzyme is distinct from the extracellular enzyme recently characterized and cloned from the same bacterial strain [C. Ogino et al., J. Biochem. 125 (1999) 263-269] and is therefore regarded as a novel prokaryotic PLD.

Bacterial Proteins↗

PubMed2001

The effect of K(atp)channel activation on myocardial cationic and energetic status during ischemia and reperfusion: role in cardioprotection.

The role of cation and cellular energy homeostasis in ATP-sensitive K(+)(K(ATP)) channel-induced cardioprotection is poorly understood. To evaluate this, rapidly interleaved(23)Na and(31)P NMR spectra were acquired from isolated rat hearts exposed to direct K(ATP)channel activation from nicorandil or pinacidil. Nicorandil attenuated ATP depletion and intracellular Na(+)(Na(+)(i)) accumulation, delayed the progression of acidosis during zero-flow ischemia and prevented ischemic contracture. The K(ATP)channel inhibitor 5-hydroxydecanoate abolished these effects. Pinacidil did not alter Na(+)(i)accumulation, ATP depletion or pH during ischemia under the conditions employed. Both agonists greatly improved the post-ischemic functional recovery. Both agonists also dramatically improved the rate and extent of the reperfusion recoveries of Na(+)(i), PCr and ATP. The Na(+)(i)and PCr reperfusion recovery rates were tightly correlated, suggesting a causal relationship. Separate atomic absorption tissue Ca(2+)measurements revealed a marked reperfusion Ca(2+)uptake, which was reduced two-fold by pinacidil. In conclusion, these results clearly indicate that while K(ATP)channel-induced metabolic alterations can vary, the functional cardioprotection resulting from this form of pharmacological preconditioning does not require attenuation of acidosis, cellular energy depletion, or Na(+)(i)accumulation during ischemia. Rather than preservation of cationic/energetic status during ischemia, the cardioprotective processes may involve a preserved capability for its rapid restoration during reperfusion. The enhanced reperfusion Na(+)(i)recovery may be enabled by the improved reperfusion cellular energy state. This accelerated Na(+)(i)recovery could play an important cardioprotective role via a potential causal relationship with the reduction of reperfusion tissue Ca(2+)uptake and resultant reperfusion injury.

Adenosine Triphosphate↗

PubMed2001

Hemispheric shape of European and Japanese brains: 3-D MRI analysis of intersubject variability, ethnical, and gender differences.

Hemispheric shape is studied using magnetic resonance imaging and 3-D reconstructions in right-handed, male and female, European and Japanese subjects. Japanese hemispheres are relatively shorter, but wider than European hemispheres. Regions of maximal intersubject variability in hemispheric shape are present in the occipital and temporal lobes in each sample. Deviations from this general pattern are found in the (i) right inferior parietal lobule (European hemispheres are more variable than Japanese), (ii) lower third of the pre- and postcentral gyri (female Japanese hemispheres are less variable than the other samples), (iii) right inferior frontal gyrus (male European hemispheres are more variable than the other samples), and (iv) polar part of the frontal lobe (female European hemispheres are less variable than the other samples). The distribution of intersubject variability between the hemispheres is less asymmetric in female than male brains. Male Japanese hemispheres are shorter but wider than female Japanese hemispheres, whereas European hemispheres show the inverse gender relations. These results demonstrate that hemispheric shape shows a considerable intersubject variability, which is not randomly distributed over the cortical surface but displays distinct regions of higher variability. Despite this intersubject variability significant interethnic- and gender-related differences in hemispheric shape are present, which may be relevant if individual brains have to be warped to a single or mean reference brain or realistic brain models are to be constructed.

Adult↗

PubMed2001

Activation reduction in anterior temporal cortices during repeated recognition of faces of personal acquaintances.

Repeated recognition of the face of a familiar individual is known to show semantic repetition priming effect. In this study, normal subjects were repeatedly presented faces of their colleagues, and the effect of repetition on the regional cerebral blood flow change was measured using positron emission tomography. They repeated a set of three tasks: the familiar-face detection (F) task, the facial direction discrimination (D) task, and the perceptual control (C) task. During five repetitions of the F task, familiar faces were presented six times from different views in a pseudorandom order. Activation reduction through the repetition of the F tasks was observed in the bilateral anterior (anterolateral to the polar region) temporal cortices which are suggested to be involved in the access to the long-term memory concerning people. The bilateral amygdala, the hypothalamus, and the medial frontal cortices, were constantly activated during the F tasks, and considered to be associated with the behavioral significance of the presented familiar faces. Constant activation was also observed in the bilateral occipitotemporal regions and fusiform gyri and the right medial temporal regions during perception of the faces, and in the left medial temporal regions during the facial familiarity detection task, which are consistent with the results of previous functional brain imaging studies. The results have provided further information about the functional segregation of the anterior temporal regions in face recognition and long-term memory.

Adult↗

PubMed2001

Different distribution of the activated areas in the dorsal premotor cortex during visual and auditory reaction-time tasks.

Sensorimotor association is an essential aspect of behavior. The dorsal part of the premotor cortex (PMd) is known to have an important role in sensorimotor association. Although it is suggested that the partially segregated groups of neurons are involved in sensorimotor association in different sensory modalities, it is not yet clear whether these groups occupy the PMd to the same or different extent. Therefore, we performed a functional magnetic resonance imaging study to compare activated regions in the PMd during simple reaction-time tasks with visual and auditory cues. Eight normal volunteers performed two simple reaction time tasks with a conventional on-off design; one is with a visual cue and the other is with an auditory cue. In both tasks, two regions in the left primary motor area (M1) (4a and 4p) and the bilateral PMd were activated. The two activated regions in the left M1 occupied the same areas in both the visual and the auditory tasks. However, in the PMd, the activated regions were situate medially during the visual task and laterally during the auditory task, along the precentral sulci. There was no overlap of significantly activated regions between two tasks, and areas specifically activated during the visual task were observed in the middle of the precentral sulci, bilaterally. The results suggest that the distribution of PMd subregions involved in sensorimotor association differ when the sensory cues are in different modalities.

Adult↗

PubMed2001

Activation in the ipsilateral posterior parietal cortex during tool use: a PET study.

The basis of perceptual assimilation of tool and hand has been considered to be in modification of body schemata, for which integration of multimodal sensory information about our body parts is required. Using positron emission tomography and H(2)(15)O, we aimed to identify brain regions that change their neural activity in association with changes in neural processing of visual and/or somatosensory information when humans use a simple tool. Normal subjects were instructed to manipulate a small graspable object with a pair of tongs or with the fingers of their right or left hand. The only site activated during manipulation with the tool, compared with the fingers, with the right hand was the lateral edge of the right intraparietal sulcus (IPS). During manipulation using the left hand with the tool, compared with using the fingers, an area in the middle part of the left IPS was activated. Areas in the contralateral hemisphere were activated during both the tool-use and the finger-use tasks compared to the control task, but there was no statistically significant difference between the tool-use and the finger-use tasks. Therefore, the results suggest that the ipsilateral posterior parietal cortex was recruited during the tool-use tasks to integrate visuosomatosensory information.

Adult↗

PubMed2001

Yeast whole-cell biocatalyst constructed by intracellular overproduction of Rhizopus oryzae lipase is applicable to biodiesel fuel production.

Yeast whole-cell biocatalysts for lipase-catalyzed reactions were constructed by intracellularly overproducing Rhizopus oryzae lipase (ROL) in Saccharomvces cerevisiae MT8-1. The gene encoding lipase from R. orvzae IFO4697 was cloned, and intracellular overproduction systems of a recombinant ROL with a pro-sequence (rProROL) were constructed. When rProROL from R. oryzae IFO4697 was produced under the control of the 5'-upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL) at 30 degrees C for 98 h by two-stage cultivation using SDC medium (SD medium with 2% casamino acids) containing 2.0% and 0.5% glucose, intracellular lipase activity reached levels up to 474.5 IU/l. These whole-cell biocatalysts were permeabilized by air-drying and used for the synthesis of methyl esters (MEs), a potential biodiesel fuel, from plant oil and methanol in a solvent-free and water-containing system. The ME content in the reaction mixture was 71 wt% after a 165-h reaction at 37 degrres C with stepwise addition of methanol. These results indicate that an efficient whole-cell biocatalyst can be prepared by intracellular overproduction of lipase in yeast cells and their permeabilization.

Amino Acid Sequence↗

PubMed2001

Development of novel whole-cell immunoadsorbents by yeast surface display of the IgG-binding domain.

The ZZ domain derived from Staphylococcus aureus, which binds to the Fc part of immunoglobulin G (IgG), was displayed on the cell surfaces of yeast Saccharomyces cerevisiae by cell-surface engineering using the C-terminal half of alpha-agglutinin under control of the 5'-upstream region of the isocitrate lyase gene from Candida tropicalis (UPR-ICL). Display of ZZ on the cell surface was confirmed by immunofluorescence microscopy. Enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA using the S. cerevisiae cells displaying ZZ detected IgG and antigen (human serum albumin) down to a concentration of 1-10 ng/ml in both cases. The detection range covered by these assay systems was wide and could be varied by adjusting the amount of cells and reaction times with horseradish peroxidase (HRP) substrate. Moreover, yeast cells displaying ZZ were successfully used for repeated affinity purification of IgG from serum. These results indicate that S. cerevisiae displaying ZZ may constitute novel and genetically renewable whole-cell immunoadsorbents widely applicable to immunoassays and affinity purification.

Animals↗

PubMed2001

Advantage of delayed whole-body FDG-PET imaging for tumour detection.

Delayed imaging that coincides with the highest uptake of fluorine-18 fluorodeoxyglucose (FDG) by tumour may be advantageous in oncological positron emission tomography (PET), where delineation of metastasis from normal tissue background is important. In order to identify the better imaging protocol for tumour detection, whole-body FDG-PET images acquired at 1 h and 2 h after injection were evaluated in 22 subjects, with a post-injection transmission scan at 90 min for attenuation correction. After visual interpretation, tumour uptake [tumour standardised uptake ratio (SUR)], normal tissue uptake (normal SUR) and tumour to background contrast (tumour SUR/normal tissue SUR) were evaluated in the images acquired at 1 h and at 2 h. Most malignant lesions, including primary lung cancer, metastatic mediastinal lymph nodes and lymphoma lesions, showed higher FDG uptake at 2 h than at 1 h. By contrast, benign lesions, with the exception of sarcoidosis, showed lower uptake of FDG at 2 h than at 1 h. Among normal tissues, the kidney, liver, mediastinum, lung, upper abdomen and left abdomen showed significant falls in FDG uptake from 1 h to 2 h. The lower abdomen, right abdomen and muscles (shoulder and thigh) showed no significant changes. Consequently, malignant lesions of the lung, mediastinum and upper abdomen showed significant increases in tumour to background contrast from 1 to 2 h. Three lesions (two lung cancers and a malignant lymphoma) that were equivocal on 1-h images became evident on 2-h images, changing the results of interpretation. All other malignant lesions were detected on 1-h images, but were clearer, with higher contrast, on 2-h images. Lesion-based sensitivity was improved from 92% (49/53) to 98% (52/53), and patient-based sensitivity from 78% (14/18) to 94% (17/18). It is concluded that delayed whole-body FDG-PET imaging is a better and more reliable imaging protocol for tumour detection.

Fluorodeoxyglucose F18↗

PubMed2001

Relationship between functional deficit and severity of experimental fast-strain injury of rat skeletal muscle.

We developed a rat model of fast-strain muscle injury to examine the relationship between functional deficit and the degree of muscle damage. A single fast strain was applied to the right plantaris (Plt) muscle of adult male rats. A tetanic contraction was induced by stimulating the sciatic nerve. Three types of strain injuries were produced by manipulating the timing of strain and contraction conditions, including applying the strain at: (1) the shortening phase (SP), (2) the full contraction phase (FCP) during tetanic contraction and (3) when the muscle is not contracting, i.e. non-contraction (NC). The contralateral Plt muscle was used as the control. Morphological and functional analysis were performed and the severity of strain injury was estimated by measuring [3H]thymidine labelling 48 h after the strain was applied. The results show a significant decrease in tension output in all three groups at 5 min after strain application (P < 0.01). The functional deficit lasted for 3 weeks in the NC group, while rats of the SP and FCP groups showed recovery 1 week after strain. Increased uptake of [3H]thymidine was similar in SP and FCP groups, but 3.5-fold higher in NC than in SP and FCP groups (P < 0.01). Histological analysis revealed an increase in the size of interstitial spaces of the muscle in NC compared to SP and FCP groups (P<0.05). Our results suggest that the contraction of muscle fibres limits the severity of connective tissue damage, and that muscle damage accompanied by disruption of the muscle connective tissue network requires a relatively longer recovery time.

Animals↗

PubMed2001

Involvement of local intercellular communication in the differentiation of zinnia mesophyll cells into tracheary elements.

The transdifferentiation of isolated mesophyll cells of zinnia (Zinnia elegans L.) into tracheary elements (TEs) has been well studied as a model of plant cell differentiation. In order to investigate intercellular communication in this phenomenon, two types of culture method were developed, in which mesophyll cells were embedded in a thin sheet of agarose gel and cultured on solid medium, or embedded in microbeads of agarose gel and cultured in liquid medium. A statistical analysis of the two-dimensional distribution of TEs in the thin-sheet cultures demonstrated their aggregation. In the microbead cultures, the frequency of TE differentiation was shown to depend on the local cell density (the cell density in each microbead): TE differentiation required local cell densities of more than 10(5) cells ml(-1). These results suggest that TE differentiation involves cell-cell communication mediated by a locally acting diffusible factor. This presumptive factor was characterized by applying a modified version of the sheet culture, which used two sheets of different cell densities, a low-density sheet and a high-density sheet. Differentiation of TEs in the former could be induced only by bringing it into contact with the latter. Insertion of a 25-kDa-cutoff membrane between the high-density and low-density sheets severely suppressed such induction of TEs in the low-density sheet while a 300-kDa-cutoff membrane suppressed induction only slightly. Insertion of agarose sheets containing immobilized pronase E or trypsin also interfered with the induction of TEs in the low-density sheets. Thus, a proteinaceous macromolecule of 25-300 kDa in molecular weight was assumed to mediate the local intercellular communication required for TE differentiation. This substance was designated "xylogen" with reference to its xylogenic activity. The time of requirement for xylogen during TE differentiation was assessed by experiments in which cells in the low-density sheet were separated from xylogen produced in the high-density sheet at various times by insertion of a 25-kDa-cutoff membrane between the two sheets, and was estimated to be from the 36th hour to the 60th hour of culture (12-36 h before visible thickening of secondary cell walls of TEs).

Asteraceae↗

PubMed2001

Gastrointestinal stromal tumor of the lesser omentum: report of a case.

We describe herein an extremely unusual case of a gastrointestinal stromal tumor (GIST) of the lesser omentum. A 45-year-old man was admitted to our hospital with an intra-abdominal mass that was subsequently misdiagnosed as a submucosal tumor of the stomach. The tumor arose from the lesser omentum and was removed without difficulty. Histologically, the tumor was composed of spindle-shaped cells with an interlacing bundle pattern, and immunohistochemical examination showed that it was positive for myeloid stem cell antigen (CD34), but negative for HHF35 and S-100 protein. These findings were consistent with a GIST lacking myogenic features and neural attributes. The patient had an uneventful postoperative course, and was free of recurrence when last seen 11 months after his operation.

Gastrointestinal Neoplasms↗

PubMed2001

Pure V1 trigeminal neuralgia caused by a cryptic trigeminal neurinoma.

Most idiopathic trigeminal neuralgias are caused by vascular compression at the root entry zone of the trigeminal nerve. (Dandy [1] and Jannetta [2]) We report a case of pure V1 trigeminal neuralgia caused by a small trigeminal neurinoma which had not been detected on preoperative neuroradiological examination.

Diagnosis, Differential↗

PubMed2001

Neural substrates for recognition of familiar voices: a PET study.

Identification of familiar people is essential in our social life. We can identify familiar people by hearing their voices as well as by viewing their faces. By measuring regional cerebral blood flow (rCBF) by positron emission tomography (PET), we identified neural substrates for the recognition of familiar voices. The brain activity during discrimination of voices of the subjects' associates and friends from those of unfamiliar people was compared with that during an analogous discrimination of their own voice from unfamiliar voices as well as during vowel discrimination. The left frontal pole, right temporal pole, right entorhinal cortex, and left precuneus were activated to a greater extent during discrimination of familiar voice than during control discriminations, suggesting that these brain regions are involved in the recognition of familiar voices. Furthermore, the adjusted values of rCBF in the left frontal pole and right temporal pole correlated with the number of subjects' correct identification of familiar voices. The present results suggest that these two regions are coactively associated with matching the currently heard voice to familiar voices in one's memory.

Adult↗

PubMed2001

Different neural systems for recognizing plants, animals, and artifacts.

The purpose of this study was to investigate functional organization in the human brain involved in the representation of knowledge regarding plants. We measured the brain activity of eight male volunteers during the recognition of visual stimuli representing plants, animals and artifacts, using positron emission tomography. The participants were presented with and were required to name silently two different images each of 15 entities belonging to three ontological categories, and 30 series of four to six digits. Marked increases in regional cerebral blood flow were found in the hippocampus and the parahippocampal areas bilaterally and the right lateral occipital cortex during the silent naming of all three categories, compared with that during the silent reading of digits. The right lateral occipital cortex was specifically activated in association with the naming of plants, and the right fusiform cortex was specifically activated in association with the naming of animals. In addition, the right temporo-occipital cortex was activated only during animals and plants, not artifacts. Our results indicate that there were a few characteristic activations for the different categories, and that entities belonging to the different categories are not necessarily represented in different locations of the brain.

Cerebral Cortex↗

PubMed2001

Study of movements of individual structures of the larynx during swallowing.

OBJECTIVE: The purpose of this study was to reveal movements of individual structures of the larynx during swallowing. METHODS: Subjects were 7 healthy adults, aged 24 to 32 years (average 27 years), who had no organic or functional disease of the pharynx and larynx and 2 adults with unilateral recurrent nerve palsy, aged 49 and 60 years, respectively. We used the Toshiba IIDR system, which is composed of an X-ray TV system and a digital image managing circuit. One-fifth diluted Omnipark 300 was used as contrast medium (lohexol), with 15 cc for each swallow. The mask image for subtraction was designated as the frame before laryngeal elevation during swallowing for subtraction. We obtained the images for observation and analysis after subtracting the mask image from continuously obtained images. These images were captured into a personal computer at 30 frames per s and thereafter frame-by-frame observation and analysis were performed by means of NIH image 1.56. RESULTS: We observed that the vocal folds underwent a series of movements during swallowing. (1) They adduct slowly and do not ascend; (2) then begin to ascend and continue adducting; whereby (3) they abduct for a moment while ascending. (4) Again, they adduct and achieve closure. (5) While maintaining closure, vocal folds elevate further to reach their maximal elevation; and (6) begin to abduct rapidly while maintaining maximal elevation. Finally, (7) they begin to descend and undergo repeated irregular abduction and adduction while descending. In examining the relationship between closure and opening at levels of the vocal fold and false vocal fold, we found that closure at the false vocal fold level precedes that at the vocal fold level and that opening at the vocal fold level precedes that at the level of the false vocal fold. CONCLUSION: Closure of the false vocal fold level appears important in the protection of the lower respiratory tract during swallowing.

Adult↗