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Biomedical subjects

A Nakamura

Publications and source records attributed to A Nakamura.

At least 127 records · Page 7Linked to original sources
PubMed2000

Assembly of smooth muscle myosin by the 38k protein, a homologue of a subunit of pre-mRNA splicing factor-2.

Smooth muscle myosin in the dephosphorylated state does not form filaments in vitro. However, thick filaments, which are composed of myosin and myosin-binding protein(s), persist in smooth muscle cells, even if myosin is subjected to the phosphorylation- dephosphorylation cycle. The characterization of telokin as a myosin-assembling protein successfully explained the discrepancy. However, smooth muscle cells that are devoid of telokin have been observed. We expected to find another ubiquitous protein with a similar role, and attempted to purify it from chicken gizzard. The 38k protein bound to both phosphorylated and dephosphorylated myosin to a similar extent. The effect of the myosin-binding activity was to assemble dephosphorylated myosin into filaments, although it had no effect on the phosphorylated myosin. The 38k protein bound to myosin with both COOH-terminal 20 and NH(2)-terminal 28 residues of the 38k protein being essential for myosin binding. The amino acid sequence of the 38k protein was not homologous to telokin, but to human p32, which was originally found in nuclei as a subunit of pre-mRNA splicing factor-2. Western blotting showed that the protein was expressed in various smooth muscles. Immunofluorescence microscopy with cultured smooth muscle cells revealed colocalization of the 38k protein with myosin and with other cytoskeletal elements. The absence of nuclear immunostaining was discussed in relation to smooth muscle differentiation.

Amino Acid Sequence↗
PubMed2000

Stable transfectants of smooth muscle cell line lacking the expression of myosin light chain kinase and their characterization with respect to the actomyosin system.

We constructed a plasmid vector having a 1.4-kilobase pair insert of myosin light chain kinase (MLCK) cDNA in an antisense direction to express antisense mRNA. The construct was then transfected to SM3, a cell line from vascular smooth muscle cells, producing a few stable transfectants. The down-regulation of MLCK expression in the transfectants was confirmed by both Northern and Western blots. The control SM3 showed chemotaxic motility to platelet-derived growth factor-BB, which was supported by lamellipodia. However, the transfectants showed neither chemotaxic motility nor developed lamellipodia, indicating the essential role of MLCK in the motility. The specificity for the targeting was assessed by a few tests including the rescue experiment. Despite this importance of MLCK, platelet-derived growth factor-BB failed to induce MLC20 phosphorylation in not only the transfectants but also in SM3. The mode in which MLCK was involved in the development of membrane ruffling is discussed with special reference to the novel property of MLCK that stimulates the ATPase activity of smooth muscle myosin without phosphorylating its light chain (Ye, L.-H., Kishi, H., Nakamura, A., Okagaki, T., Tanaka, T., Oiwa, K., and Kohama, K. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 6666-6671).

Actomyosin↗
PubMed2000

Studies on the mechanisms of tumorigenesis induced by polyetherurethane in rats: production of superoxide, tumor necrosis factor, and interleukin 1 from macrophages cultured on different polyetherurethanes.

Interaction of macrophages (Mos) with polyetherurethane (PEU) was investigated to clarify the role of the Mos in the early stage of tumorigenesis of PEUs. As for the inflammatory cytokines produced from Mos, the amount of tumor necrosis factor (TNF) produced on three PEUs (PU-4, 6, and 8) was similar, but less than that on a control glass dish. Interleukin 1 (IL-1) production levels on the PEUs, especially on PU-6, also decreased in comparison with the glass dish. Superoxide (O(2)(-)) release from Mos on the PEUs was similar or more than that on the glass dishes within 24 h incubation. In particular, the O(2)(-) release on PU-6, possessing the lowest tumorigenic potential in vivo, reached the highest level among the PEUs tested. To clarify the causes that enhanced O(2)(-) release from Mos, the methanol extracts of the three PEUs and chemicals that constitute the PEUs were tested. The extracts and 1,4-butanediol did not show an effect on O(2)(-) release. However, 4, 4'-di(ethoxycarboamide) diphenylmethane, a model hard segment of PEU, was remarkably enhanced, whereas poly(tetramethyleneoxide), a soft segment of PEU, reduced the O(2)(-) release in a dose-dependent manner. From these results, although it is still unclear what affects different types of O(2)(-) production, lower tumorigenic potentials of PU-6 may be caused by O(2)(-) release from Mos, which play an important role in the tumoricidal process. Further, the low IL-1 production on PU-6 suggests that PU-6 suppresses inflammatory responses other than O(2)(-) production in vivo, resulting in lower tumorigenic potentials. On the other hand, TNF production was almost similar when Mos were cultured on three PEUs, suggesting TNF did not play a role in the different tumorigenic potentials of PEUs.

Animals↗
PubMed2000

Beta2-adrenoceptor agonist suppresses tumour necrosis factor production in rat mesangial cells.

This study aimed to investigate the time-course of the effect of beta2-adrenoceptor stimulation with terbutaline on lipopolysaccharide (LPS)-induced tumour necrosis factor(TNF)-alpha production in rat mesangial cells. Cells were cultured from 0-24 h in the presence of LPS (1 microg/ml) and/or terbutaline (10(-7)-10(-8) mol/l). After 1 h of incubation, terbutaline inhibited TNF-alpha protein release as well as transcription and translation of TNF-alpha and mitogen activated protein kinase (MAPK, p42/p44) activity. At 3 h, terbutaline enhanced intracellular cAMP but suppressed TNF-alpha release and transcription. By 24 h, whereas terbutaline was no longer influencing transcription or translation, TNF-alpha release remained depressed which correlated with an increase in supernatant interleukin (IL)-6. Terbutaline did not affect the LPS-induced IL-10 produced in the cell. These findings indicate that beta2-adrenoceptor stimulation during an LPS challenge prevented TNF-alpha production as a consequence of MAPK inhibition and enhanced cAMP generation, which at a later stage was associated with an anti-inflammatory effect of IL-6.

Adrenergic beta-Agonists↗
PubMed2000

Ataxin-3 is translocated into the nucleus for the formation of intranuclear inclusions in normal and Machado-Joseph disease brains.

Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 (SCA3) is one of the dominantly inherited cerebellar ataxias. The gene responsible for the disease, a novel gene of unknown function, encodes ataxin-3 containing a polyglutamine stretch. Although it has been known that ataxin-3 is incorporated into neuronal intranuclear inclusions (NIIs) in neurons of affected regions, the relationship between NII formation and neuronal degeneration still remains uncertain. In the present study we show two different conditions in which ataxin-3 is recruited into the nucleus and suggest a process to form nuclear inclusions. In normal brains, wild-type ataxin-3 localizes within the ubiquitin-positive nuclear inclusion, the Marinesco body, indicating that ataxin-3 is recruited into the nuclear inclusion even in the absence of pathologically expanded polyglutamine. In MJD/SCA3 brains, immunohistochemical analyses with anti-ataxin-3 antibody, anti-ubiquitin antibody, and monoclonal antibody 1C2 known to recognize expanded polyglutamine revealed differences in frequency and in diameter among NIIs recognized by each antibody. These results were confirmed in the same inclusions by double immunofluorescent staining, suggesting that expanded ataxin-3 forms a core, thereby recruiting wild-type ataxin-3 into the nucleus around the core portion, and then followed by activation of the ubiquitin/ATP-dependent pathway. Recruitment of ataxin-3 into the nucleus and formation of nuclear inclusion under two different conditions suggest that ataxin-3 may be translocated into the nucleus under certain conditions stressful on neuronal cells such as aging and polyglutamine neurotoxicity.

Ataxin-3↗
PubMed2000

Inhibitory effects of ovoglobulins on bacillary necrosis in larvae of the pacific oyster, Crassostrea gigas.

In order to develop an alternative method to antibiotics for preventing bacillary necrosis in bivalve mollusc larvae, we examined the effects of ovoglobulins (proteins derived from the whites of hens' eggs) on the survival of larvae of the Pacific oyster Crassostrea gigas. The pathogenic Vibrio tubiashii (ATCC 19106) was used to infect larvae of the Pacific oyster. V. tubiashii showed strong pathogenicity to oyster larvae, causing 100% mortality after experimental exposure for 24 h at a concentration of 10(5) cfu (colony-forming units)/ml. In contrast, the addition of ovoglobulins at a concentration of 10 microg/ml to larval oysters, challenged with V. tubiashii at 10(5) cfu/ml, led to a marked increase in larval survival of 96.5% at 24 h after infection. The V. tubiashii culture supernatant was also shown to be pathogenic to larval oysters; however, its pathogenicity was completely inhibited by the addition of 10 microg/ml of ovoglobulins. Larval oysters infected by V. tubiashii showed typical symptoms of bacillary necrosis including anomalous swimming and detachment of cilia and/or vela. In contrast, live larvae were actively motile, and their cilia and vela were not necrotized in the ovoglobulins-added group. The addition of ovoglobulins clearly suppressed the growth of V. tubiashii in gelatin-sea water broth, but the number of viable V. tubiashii 24 h after incubation did not decrease to the initial dose level. Findings obtained in this study indicate that ovoglobulins almost completely protect larval oysters from V. tubiashii infection by nonbactericidally inhibiting the growth of V. tubiashii without affecting survival of the oysters.

Animals↗
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

Extrastriatal mean regional uptake of fluorine-18-FDOPA in the normal aged brain--an approach using MRI-aided spatial normalization.

The aim of this study was to define the mean regional 6-[(18)F]fluoro-l-dopa (FDOPA) uptake rate constant (K(i)) values in the striatal and extrastriatal regions of the brain of normal subjects using magnetic resonance imaging (MRI)-aided spatial normalization of the FDOPA K(i) image and using automatic region of interest (ROI) analysis. Dynamic three-dimensional FDOPA positron emission tomography (PET) and three-dimensional magnetic resonance (MR) images were acquired in 13 aged normal subjects. The FDOPA add image and the K(i) image of each subject were transformed into standard stereotactic space with the aid of individual coregistered MR image. The mean regional K(i) values of the striatal and extrastriatal regions before normalization were compared with the respective values after normalization. Then automatic ROI analysis was performed on the MRI-aided spatially normalized K(i) images of the 13 normal subjects. The K(i) values on original images and those on spatially normalized images were in good agreement, indicating that the spatial normalization technique did not change the regional K(i) values appreciably. Automatic ROI analysis of the spatially normalized FDOPA K(i) images of the normal subjects, showed high K(i) values in ventral and dorsal regions of the midbrain, amygdala, hippocampus, and medial prefrontal cortex, in addition to caudate nucleus and putamen, which correspond to the dopaminergic projections in the brain. Spatial normalization technique helped to establish a database of FDOPA K(i) images of normal subjects and high K(i) values were observed widely besides striatal regions corresponding to the dopaminergic projections in the brain.

Aged↗
PubMed2000

Video-assisted mediastinoscopic resection of a thymic cyst by sternum lifting.

We performed mediastinoscopic surgery on two patients with a thymic cyst using a new instrument by sternum-lifting with tape. The merits of this technique are that a resection of the thymus and cyst can be achieved without a midline sternotomy, and under bilateral ventilation, and in cases with pleural adhesion. This technique can be applicable to most benign thymic lesions including thymectomy for myasthenia gravis and to selected cases with a malignant thymic tumor.

Aged↗
PubMed2000

Tau-positive neurons in corticobasal degeneration and Alzheimer's disease--distinction by thiazin red and silver impregnations.

Thiazin red (TR), a fluorochrome that has an affinity to fibrillary structures such as neurofibrillary tangles (NFTs) or senile plaques, was utilized to investigate assembly of tau protein into fibrils in tau-immunopositive neocortical neurons of corticobasal degeneration (CBD) and of Alzheimer's disease (AD). Double fluorescence with anti-paired helical filament monoclonal antibody (AT8) and TR was followed by either the Gallyas or Bodian silver impregnation method, which enabled a comparison of the staining features by three different methods on the same neuron. NFTs of AD were uniformly stained by TR and Gallyas method. Most of tau-immunopositive neurons of CBD were similarly stained by Gallyas method but barely or only weakly by TR or Bodian method, suggesting that tau in neocortical neurons of CBD is less liable to form fibrillary structures than in those of AD, easily distinguishable by TR staining. Clarifying the process of tau assembly using this fluorochrome will give a clue to understanding mechanisms of tau deposition, which may be different in various neurological disorders.

Aged↗
PubMed2000

Dual enhancement of double immunofluorescent signals by CARD: participation of ubiquitin during formation of neurofibrillary tangles.

Amplification with catalyzed reporter deposition (CARD) greatly enhances peroxidase signals, which has been utilized to amplify immunohistochemical labelings including fluorochromes. Here we describe a strategy to amplify each of two immunofluorescent signals without crosstalk on double-stained histological sections from human autopsied brains with Alzheimer's disease (AD). One of the two primary antibodies (anti-Abeta or anti-PHF-tau) was probed by a species-specific secondary antibody conjugated with horseradish peroxidase (HRP), which was visualized by FITC-labeled tyramide. After inactivation of HRP, the other primary antibody was probed by another species-specific secondary antibody conjugated with HRP. Amplification with biotinylated tyramide was followed by streptavidin-conjugated Cy-5, which specifically labeled the latter epitope. It was found that Abeta and PHF-tau were localized to senile plaques and neurofibrillary tangles (NFTs), respectively, which verified lack of crosstalk on the double-stained section. Localization of ubiquitin and PHF-tau was looked for at higher magnification in NFT-bearing neurons. Although these two epitopes were colocalized in some neurons, ubiquitin was not always present in PHF-tau positive NFTs. Discrepancy between PFH-tau and ubiquitin, verified inter- and intracellularly, may represent different stages of NFT formation. This is the first report of successful CARD amplification of two different fluorescent signals on double-labeling immunohistochemistry, which is now proved to be powerful in detecting epitopes in relation to AD-related lesions. Improved intensity over tenfold of the two fluorescent signals without crosstalk will expand the application of the multilabeling method with fluorochromes.

Alzheimer Disease↗
PubMed2000

Post-ischemic intraportal adenosine administration protects against reperfusion injury of canine liver.

Although adenosine has been postulated to inhibit ischemia-reperfusion injury in various tissues, its in vivo cytoprotective mechanism is not fully known. The aim of this study was to determine the effect of intraportally infused adenosine on reperfusion injury in the canine liver. Two h ischemia and reperfusion of the liver were induced in beagle dogs by clamping the portal triad. Either adenosine or saline was infused in the portal vein after reperfusion for 60 min. Levels of serum aspartate aminotransferase and alanine aminotransferase and the survival of animals were examined. Hepatic levels of protein carbonyls and glutathione were also measured, as markers of oxidative stress. One h after reperfusion, the liver was perfused with nitroblue tetrazolium and the formation of formazan was observed to evaluate superoxide formation. Twenty-four h after reperfusion, 100% of animals in the adenosine group and 33% of animals in the control group survived. Adenosine significantly decreased the reperfusion-induced increase in serum levels of aspartate aminotransferase and alanine aminotransferase. Adenosine also suppressed the formation of protein carbonyls and the decrease in glutathione levels. Histologically, neutrophil infiltration, superoxide formation, and apoptosis were decreased by adenosine. These results suggest that intraportally infused adenosine attenuates reperfusion injury of the liver, presumably by suppressing the activation of neutrophils and oxidative stress.

Adenosine↗
PubMed2000

Effect of heat treatment of poly(L-lactide) on the response of osteoblast-like MC3T3-E1 cells.

Poly(L-lactide) (PLLA) products are molded by heat extrusion. These treatments may change chemical properties and biological response of the PLLA to cells. In this study, the effect of heat treatment of PLLA on osteoblast proliferation and differentiation was examined in vitro. Osteoblast-like MC3T3-E1 cells were cultured for 2 weeks on the PLLA subjected to various heating temperature and time combinations. The protein, DNA, and hydroxyproline (HYP) contents and alkaline phosphatase (ALP) activity of cells cultured on the untreated (non-heated) PLLA with a weight average molecular weight (Mw) of 1,000,000 (high Mw PLLA) were not significantly different from those of cells cultured on glass. The activation of osteoblast differentiation by the high Mw PLLA was weak. In contrast, increases in ALP activity and HYP content were found for cells cultured on the PLLA heated at a high temperature of 200 or 250 degrees C. Heat treatment of high Mw PLLA increased differentiation of MC3T3-E1 cells cultured upon it. Significant degradation of PLLA (decrease in molecular weight and increase in molecular weight distribution) were observed following heat treatment. The Mw of PLLA decreased from 1,000,000 to below 20,000, and 14.4 microg of L-lactic acid was released from 10 mg of PLLA by heating at 250 degrees C. Therefore, the effect of low Mw chemicals, which were expected to be the degradation products of high Mw PLLA after heat treatment, on MC3T3-E1 cell activities was examined. Increases in the protein, DNA and HYP amounts and ALP activity for cells cultured with L-lactide or L-lactic acid were observed at 100 microg/ml, but not at 10 microg/ml. When the cells were cultured on the low Mw PLLA (Mw 20,000), their biological parameters also increased. Twelve micrograms of L-lactic acid released from 10 mg of the low Mw PLLA during 2 weeks incubation. The concentration of L-lactic acid in the incubation solution of low Mw PLLA or heat-treated PLLA was too small to cause cell activation. These results suggested that increases in osteoblast differentiation on the heat-treated PLLA was not to due to soluble degradation chemicals, such as L-lactic acid, rather than the remaining low Mw PLLA.

Absorbable Implants↗
PubMed2000

The somatosensory evoked magnetic fields.

Averaged magnetoencephalography (MEG) following somatosensory stimulation, somatosensory evoked magnetic field(s) (SEF), in humans are reviewed. The equivalent current dipole(s) (ECD) of the primary and the following middle-latency components of SEF following electrical stimulation within 80-100 ms are estimated in area 3b of the primary somatosensory cortex (SI), the posterior bank of the central sulcus, in the hemisphere contralateral to the stimulated site. Their sites are generally compatible with the homunculus which was reported by Penfield using direct cortical stimulation during surgery. SEF to passive finger movement is generated in area 3a or 2 of SI, unlike with electrical stimulation. Long-latency components with peaks of approximately 80-120 ms are recorded in the bilateral hemispheres and their ECD are estimated in the secondary somatosensory cortex (SII) in the bilateral hemispheres. We also summarized (1) the gating effects on SEF by interference tactile stimulation or movement applied to the stimulus site, (2) clinical applications of SEF in the fields of neurosurgery and neurology and (3) cortical plasticity (reorganization) of the SI. SEF specific to painful stimulation is also recorded following painful stimulation by CO(2) laser beam. Pain-specific components are recorded over 150 ms after the stimulus and their ECD are estimated in the bilateral SII and the limbic system. We introduced a newly-developed multi (12)-channel gradiometer system with the smallest and highest quality superconducting quantum interference device (micro-SQUID) available to non-invasively detect the magnetic fields of a human peripheral nerve. Clear nerve action fields (NAFs) were consistently recorded from all subjects.

Brain Mapping↗
PubMed2000

Antibiotic resistance among recent clinical isolates of Haemophilus influenzae in Japanese children.

From January 1997 to July 1999, a total of 867 isolates of Haemophilus influenzae were recovered in the microbiology laboratory of Chiba Children's Hospital. The overall prevalence of beta-lactamase production was 12.8%. Ampicillin-MICs for all of the 111 beta-lactamase-producing isolates was > or =4 microg/ml. A total of 26 beta-lactamase-negative isolates (3.4% of all beta-lactamase-negative isolates and 3.0% of all isolates) were found to be resistant to ampicillin. The prevalence of beta-lactamase negative ampicillin-resistant strains (BLNAR) increased remarkably to 8.9% during the last 7-month period. It is noteworthy that the MICs not only of penicillins but also of cephems for BLNAR were significantly higher than those for ampicillin-susceptible isolates. Eight beta-lactamase-producing isolates of H. influenzae (7.2% of all beta-lactamase-producing isolates) were resistant to amoxicillin-clavulanate (AMPC/CVA). Consequently, the overall resistance to ampicillin was 15.8%, and that to AMPC/CVA was 3.0%. The results of this study corroborate the findings of previous investigators in the US (Doern et al., 1997) regarding the emergence of BLNAR and beta-lactamase-producing AMPC/CVA-resistant strains (BLPACR) of H. influenzae. Continued monitoring of susceptibility trends will be required to guide appropriate chemotherapy.

Anti-Bacterial Agents↗
PubMed2000

Orally active cephalosporins. Part 2: synthesis, structure-activity relationships and oral absorption of cephalosporins having a C-3 pyridyl side chain.

A series of 7beta-[(Z)-2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamid o]cephalosporins having a pyridine ring connected through various spacer moieties at the C-3 position was designed and synthesized and evaluated for antibacterial activity and oral absorption in rats. All compounds showed potent antibacterial activity against Staphylococcus aureus, whereas antibacterial activity against gram-negative bacteria was markedly influenced by the spacer moiety between the pyridine and cephem nucleus. Oral absorption was influenced by the position of the pyridine nitrogen as well as by the spacer moiety. Among these compounds, FR86830 (14), having a 4-pyridylmethylthio moiety at the C-3 position, showed the most well balanced activity and moderate oral absorption.

Administration, Oral↗
PubMed2000

VASA mediates translation through interaction with a Drosophila yIF2 homolog.

The Drosophila gene vasa (vas) encodes an RNA-binding protein required for embryonic patterning and germ cell specification. In vas mutants, translation of several germline mRNAs is reduced. Here we show that VAS interacts directly with the Drosophila homolog of yeast translation initiation factor 2, encoded by a novel gene, dIF2. Embryos produced by vas/+; dIF2/+ females have pattern defects and fewer germline progenitor cells, indicating a functional interaction between endogenous vas and dIF2 activities. Mutations in other translation initiation factors do not enhance the vas phenotype, suggesting that dIF2 has a particular role in germ plasm function. We conclude that VAS regulates translation of germline mRNAs by specific interaction with dIF2, an essential factor conserved from bacteria to humans.

Amino Acid Sequence↗