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Biomedical subjects

T Yagi

Publications and source records attributed to T Yagi.

At least 37 records · Page 2Linked to original sources
PubMed2002

Quantitative assessment of harmonic power Doppler myocardial perfusion imaging with intravenous Levovist in patients with myocardial infarction: comparison with myocardial viability evaluated by thallium-201 single-photon emission computed tomography and coronary flow reserve.

BACKGROUND: Intravenous myocardial contrast echocardiography with harmonic power Doppler imaging is a novel technique for assessing myocardial perfusion. AIMS: The aim of this study was to quantitatively assess myocardial perfusion by harmonic power Doppler imaging in patients with a previous myocardial infarction and compare myocardial contrast echocardiography results with myocardial viability evaluated by thallium-201 single-photon emission computed tomography ((201)Tl-SPECT) and the results of Doppler flow measurement of coronary flow velocity reserve. METHODS: Twenty-three patients with anterior myocardial infarction who were scheduled for adenosine stress (201)Tl-SPECT underwent myocardial contrast echocardiography with harmonic power Doppler imaging. Harmonic power Doppler imaging was performed at rest and during adenosine infusion (0.15 mg/kg/min) using an intravenous infusion of Levovist. The peak colour pixel intensity ratios of the risk area to the control area were used for quantitative analysis of myocardial perfusion by harmonic power Doppler imaging. Coronary blood flow velocity was measured using Doppler-tipped guidewire in the distal portion of left anterior descending artery and coronary flow velocity reserve was calculated. RESULTS: In patients with myocardial viability assessed by (201)Tl-SPECT, pixel intensity ratios both at rest and during hyperaemia were significantly higher compared with those in patients without myocardial viability (at rest: 0.62 +/- 0.28 vs 0.37 +/- 0.17, P=0.038, during hyperaemia 0.72 +/- 0.19 vs 0.40 +/- 0.18, P=0.003). Coronary flow velocity reserve was significantly different between two groups (2.35 +/- 0.43 vs 1.49 +/- 0.53, P <0.01). CONCLUSIONS: Quantitative assessment of microvascular integrity by harmonic power Doppler imaging corresponds to the evaluation of the microcirculation by coronary flow velocity reserve.

Adenosine↗
PubMed2001

Direct interaction between a membrane domain subunit and a connector subunit in the H(+)-translocating NADH-quinone oxidoreductase.

When Paracoccus denitrificans membranes were treated with a crosslinker, m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), a cross-linked product of M(r) approximately 31 kDa was found which reacted with antibodies against the hydrophobic subunit Nqo7 and the connector subunit Nqo6. NaI treatment of the Paracoccus membranes before, but not after, the crosslinking step prevented the formation of the 31 kDa band. When Nqo7 and Nqo6 were coexpressed in Escherichia coli, both subunits were located in the membrane fraction. MBS treatment of the E. coli membranes generated the 31 kDa band as in the Paracoccus membranes. These results indicate that Nqo7 interacts with probable N2-binding Nqo6.

Antibodies↗
PubMed2001

Assessment of the developmental totipotency of neural cells in the cerebral cortex of mouse embryo by nuclear transfer.

When neural cells were collected from the entire cerebral cortex of developing mouse fetuses (15.5-17.5 days postcoitum) and their nuclei were transferred into enucleated oocytes, 5.5% of the reconstructed oocytes developed into normal offspring. This success rate was the highest among all previous mouse cloning experiments that used somatic cells. Forty-four percent of live embryos at 10.5 days postcoitum were morphologically normal when premature and early-postmitotic neural cells from the ventricular side of the cortex were used. In contrast, the majority (95%) of embryos were morphologically abnormal (including structural abnormalities in the neural tube) when postmitotic-differentiated neurons from the pial side of the cortex were used for cloning. Whereas 4.3% of embryos cloned with ventricular-side cells developed into healthy offspring, only 0.5% of those cloned with differentiated neurons in the pial side did so. These facts seem to suggest that the nuclei of neural cells in advanced stages of differentiation had lost their developmental totipotency. The underlying mechanism for this developmental limitation could be somatic DNA rearrangements in differentiating neural cells.

Animals↗
PubMed2001

Interaction of the SH2 domain of Fyn with a cytoskeletal protein, beta-adducin.

Fyn is a Src family tyrosine kinase expressed abundantly in neurons and believed to have specific functions in the brain. To understand the function of Fyn tyrosine kinase, we attempted to identify Fyn Src homology 2 (SH2) domain-binding proteins from a Nonidet P-40-insoluble fraction of the mouse brain. beta-Adducin, an actin filament-associated cytoskeletal protein, was isolated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry. beta-Adducin was tyrosine phosphorylated by coexpression with wild type but not with a kinase-negative form of Fyn in COS-7 cells. Cell staining analysis showed that coexpression of beta-adducin with Fyn induced translocation of beta-adducin from the cytoplasm to the periphery of the cells where it was colocalized with actin filaments and Fyn. These findings suggest that tyrosine-phosphorylated beta-adducin associates with the SH2 domain of Fyn and colocalizes under plasma membranes.

Amino Acid Sequence↗
PubMed2001

Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging.

Detection of protein-protein interactions is a fundamental step to understanding gene function. Here we report a sensitive and rapid method for assaying protein-protein interactions at the single-molecule level. Protein molecules were synthesized in a cell-free translation system in the presence of Cy5-puro, a fluorescent puromycin, using mRNA without a stop codon. The interaction of proteins thus prepared was visualized using a single-molecule imaging technique. As a demonstration of this method, a motor protein, kinesin, was labeled with Cy5-puro at an efficiency of about 90%, and the processive movement of kinesin along microtubules was observed by using total internal reflection microscopy. It took only 2 h from the synthesis of proteins to the functional analysis. This method is applicable to the functional analysis of various kinds of proteins.

Adenosine Triphosphate↗
PubMed2001

Lack of complex I activity in human cells carrying a mutation in MtDNA-encoded ND4 subunit is corrected by the Saccharomyces cerevisiae NADH-quinone oxidoreductase (NDI1) gene.

The gene for the single subunit, rotenone-insensitive, and flavone-sensitive internal NADH-quinone oxidoreductase of Saccharomyces cerevisiae (NDI1) can completely restore the NADH dehydrogenase activity in mutant human cells that lack the essential mitochondrial DNA (mtDNA)-encoded subunit ND4. In particular, the NDI1 gene was introduced into the nuclear genome of the human 143B.TK(-) cell line derivative C4T, which carries a homoplasmic frameshift mutation in the ND4 gene. Two transformants with a low or high level of expression of the exogenous gene were chosen for a detailed analysis. In these cells the corresponding protein is localized in mitochondria, its NADH-binding site faces the matrix compartment as in yeast mitochondria, and in perfect correlation with its abundance restores partially or fully NADH-dependent respiration that is rotenone-insensitive, flavone-sensitive, and antimycin A-sensitive. Thus the yeast enzyme has become coupled to the downstream portion of the human respiratory chain. Furthermore, the P:O ratio with malate/glutamate-dependent respiration in the transformants is approximately two-thirds of that of the wild-type 143B.TK(-) cells, as expected from the lack of proton pumping activity in the yeast enzyme. Finally, whereas the original mutant cell line C4T fails to grow in medium containing galactose instead of glucose, the high NDI1-expressing transformant has a fully restored capacity to grow in galactose medium. The present observations substantially expand the potential of the yeast NDI1 gene for the therapy of mitochondrial diseases involving complex I deficiency.

Binding Sites↗
PubMed2001

N-Methyl-D-aspartate-induced c-Fos expression is enhanced in the forebrain structures related to emotion in Fyn-deficient mice.

Fyn tyrosine kinase is thought to be involved in the control of neuronal intracellular signal transduction elicited by neurotransmitter stimulation. Emotional disorders, such as fearfulness in Fyn-deficient mice, prompted us to investigate the neural mechanisms that lead to defective emotional expression by using functional neuroanatomical methods. In order to examine the reactivity of a specific neural network to excitatory neurotransmitter administration, we mapped the distribution of c-Fos-immunoreactive neurons after administering N-methyl-D-aspartate (NMDA) to control and mutant mice at the subthreshold dose for seizure induction. The induction of neuronal c-Fos-immunoreactivity by NMDA was enhanced in the Fyn-deficient mice, and there was a much greater increase in immunopositive neurons in certain well-defined areas, such as the amygdaloid medial nuclear subdivisions, hypothalamic paraventricular nucleus, and midbrain periaqueductal gray, of the mutant. NMDA-induced c-Fos expression was attenuated by pretreatment with D-(-)-2-amino-5-phosphonovaleric acid, a competitive NMDA antagonist, both in the control and the mutant mice. These findings suggest that the excitability of the projection system from the amygdala to the hypothalamus and midbrain, the main pathways of emotional expression, is enhanced in Fyn-deficient mice.

Amygdala↗
PubMed2001

Identification of the protein product of the Coch gene (hereditary deafness gene) as the major component of bovine inner ear protein.

In order to better understand the cause of hereditary hearing impairment, we have performed a proteomic analysis of the inner ear proteins using two-dimensional gel electrophoresis. In the process of analysis, we have found very unique properties of the bovine homologue of the human COCH gene product. The COCH gene is responsible for one of the hereditary hearing impairments, DFNA9, and was recently suggested to be a possible genetic factor contributing to Ménière's disease. The Coch protein constitutes 70% of bovine inner ear proteins and is composed of 16 different protein spots, with charge and size heterogeneity. Heterogeneity of this protein suggests that the Coch gene is processed in several ways, at the transcriptional and/or posttranslational level. Much knowledge has accumulated about the hereditary hearing impairment genes; however, little research has been done regarding the protein products of those genes. This is the first report to characterize the Coch protein. Study of the Coch protein might provide more information on the mechanism of hearing and vestibular disorders.

Amino Acid Sequence↗
PubMed2001

The effect of nitric oxide production by sinusoidal endothelial cells on preservation injury during cold ischemia.

Multifactorial elements are responsible for preservation and reperfusion injury in liver allografts. Sinusoidal endothelial cells (SECs) are a primary target of cold preservation injury of the liver. We examined the correlation between nitric oxide (NO) production by SECs and their injury during cold preservation. SECs were isolated from rat livers and preserved in either Euro-Collins (EC) or University of Wisconsin (UW) solution. Injury to the SECs was more severe when preserved in the EC solution than in the UW solution during cold ischemia. In addition, NO production by SECs was found to be proportionate to the cell injury. Cell viability was not improved by the addition of NO inhibitor, L-NMMA. Further, NO inhibitor was detrimental to the SECs in a 24-h preservation in UW solution. LDH release by SECs preserved in UW solution supplemented with L-NMMA was 11.10+/-2.03 IU/l, while that in UW solution alone was 3.70+/-0.70 IU/l (P<0.01). Together, our results suggest that NO protects SECs during cold preservation and that NO from SECs may have beneficial effects on the liver during cold ischemia.

Journal Article↗
PubMed2001

Two novel CNRs from the CNR gene cluster have molecular features distinct from those of CNR1 to 8.

Cadherin-related neuronal receptor (CNR) family proteins are known as synaptic cadherins and Reelin receptors. Here we have identified two novel mouse CNR genes, CNRc1 and CNRc2, orthologues of human protocadherin (Pcdh) alpha-c1 and Pcdhalpha-c2, respectively. While the variable large exons of CNRc1 and c2 contain six conserved extracellular cadherin repeats (EC1-6) and are linked to the constant exons, both contain several molecular features distinct from CNR1-8. CNRc1 and c2 lack the Arg-Gly-Asp (RGD) sequence that is conserved in the EC1 of CNR1-8, which is necessary for binding to Reelin. The present studies confirm that CNRc1 and c2 failed to immunoprecipitate with Reelin. In addition, the regulation of novel CNR expression patterns during brain development is slightly different from that of CNR1. The identification of these new CNR genes characterized by their distinct extracellular function and expression is indicative of the novel diversity of the processes of brain structuring and synapse regulation.

Alternative Splicing↗
PubMed2001

Immunohistochemical investigation of caspase-1 and effect of caspase-1 inhibitor in delayed neuronal death after transient cerebral ischemia.

The localization of caspase-1 protein, interleukin-1beta (IL-1beta)-converting enzyme, was immunohistochemically examined in the hippocampal CA-1 subfield by a transient occlusion of bilateral common carotid arteries in Mongolian gerbils. Immunoreactivities for caspase-1 were found in microglias, astrocytes, endothelial cells of capillaries and some non-pyramidal neurons. Immunopositive microglias increased in number from 3 days until 7 days from the transient ischemia, and astrocytes also increased in number from 3 days until 28 days. At the electron microscopic level, caspase-1 immunoreaction endproducts were associated with Golgi apparatus in glial cells, endothelial cells of blood vessels and non-pyramidal neurons. The delayed neuronal death of CA-1 pyramidal cells was significantly protected by the treatment of specific caspase-1 inhibitor (Ac-WEHD-CHO) or broad caspase family inhibitor (z-VAD-FMK). Cell death was protected in a dose dependent manner by the former by 43-57%, and by the latter by 66-91% when injected at 1 and 10 microg, respectively. On the other hand, the protective effect of specific caspase-3 inhibitor (Ac-DMQD-CHO) was less significant at higher dose (10 microg) by 33% (P<0.05), and not detectable at lower dose (1 microg) by 13% (P=0.27). Furthermore, a significant decrease of microglias and astrocytes was found in the CA-1 as well as the reduction of IL-1beta and caspase-1 immunoreactivities by the treatment of Ac-WEHD-CHO. Extravasation of serum albumin was also extremely reduced by this treatment. These findings suggest that the inhibition of caspase-1 activity ameliorates the ischemic injury by inhibiting the activity of IL-1beta.

Amino Acid Chloromethyl Ketones↗
PubMed2001

Myelin basic protein is necessary for the regulation of myelin-associated glycoprotein expression in mouse oligodendroglia.

Myelin-associated glycoprotein (MAG) has currently been proposed to be a possible igniter of myelination in the central nervous system. We propose here that myelin basic protein (MBP) is prerequisite, which is more diffusely expressed than MAG in early stages of myelination. MBP-deficient mice show significant reduction of MAG-positive immature oligodendroglia at early stages of myelination, although significant increase of such cells is noted in the adult. MBP/Fyn-double deficient adult mice also show moderate increase of the same type of immature glia, but to a lesser degree than MBP-deficiency alone. The present study suggests that MBP is required for the regulation of MAG expression in oligodendroglia, involving Fyn therein. We discuss the possibility that hitherto unknown molecule, not MAG, may be in responsible for the ignition of the myelination.

Animals↗
PubMed2001

Influence of humoral immunoreaction on hepatic nonparenchymal cells in ex situ xenoperfused rat livers.

BACKGROUND: The influence of xenogeneic humoral immunoreaction on hepatic nonparenchymal cells (NPCs) was evaluated ex situ in xenoperfused rat livers. METHODS: Isolated rat livers were perfused via the portal vein (PV) for 240 min. The perfusates consisted of fresh rat blood (group 1), fresh human blood (group 2), and fresh human blood containing 5 microg/mL soluble complement receptor type 1 (Group 3). RESULTS: Deposition of human IgM and C(5b-9) complement was observed in group 2, although only human IgM deposition was detected in group 3. Portal vein pressure in group 2 rose drastically during the first 10 min. Creatine kinase BB component gradually increased in all groups, followed by an elevation in alanine aminotransferase and both parameters were significantly higher in group 2 than in groups 1 and 3. In group 2, platelet thrombi in the peripheral PVs and periportal hemorrhage were observed after 10 min, and massive necrosis around the central veins after 240 min; these changes were not observed in group 1 or 3. Production of tumor necrosis factor alpha and alpha interferon and expression of intercellular adhesion molecule 1 (ICAM-1) were lower in group 2 than in groups 1 and 3. In group 2, there were negative areas for ICAM-1 and tumor necrosis factor alpha staining around the central veins after 240 min, which were consistent with necrotic areas. CONCLUSIONS: In xenoperfused rat livers, humoral mediators initially caused the disturbance of microcirculation, which would induce long ischemia in the pericentral areas, resulting in massive necrosis. NPC necrosis may be responsible for less production of cytokines and adhesion molecules in the xenoperfused livers.

Alanine Transaminase↗
PubMed2001

Isolation of Bacillus subtilis (chungkookjang), a poly-gamma-glutamate producer with high genetic competence.

A bacterium with high poly-gamma-glutamate (PGA) productivity was isolated from the traditional Korean seasoning, Chung-Kook-Jang. This bacterium could be classified as a Bacillus subtilis, but sporulation in culture was infrequent in the absence of Mn2+. It was judged to be a variety of B. subtilis and designated B. subtilis (chungkookjang). L-Glutamate significantly induced PGA production, and highly elongated PGAs were synthesized. The volumetric yield reached 13.5 mg ml(-1) in the presence of 2% L-glutamate. The D-glutamate content was over 50% in every PGA produced under the conditions used. During PGA production, glutamate racemase activity was found in the cells, suggesting that the enzyme is involved in the D-glutamate supply. Molecular sizes of PGAs were changed by the salt concentration in the medium; PGAs with comparatively low molecular masses were produced in culture media containing high concentrations of NaCl. B. subtilis (chungkookjang) harbors no plasmid and is the first B. subtilis strain reported with both naturally high PGA productivity and high genetic competence.

Alanine Transaminase↗
PubMed2001

Expression of myeloperoxidase and cochlear dysfunction in the lipopolysaccharide-treated guinea pig.

In this study, the effect of endotoxin on the guinea pig cochlea was examined by electrophysiology and immunohistochemistry. Bacterial lipopolysaccharide (LPS) was injected into the middle ear transtympanically. Electrocochleograms were measured immediately and 48 h after the injection with an electrode inserted into the facial canal. After the electrophysiological measurement, the animals were killed with an intracardiac perfusion of fixative and the temporal bones were removed and processed for immunohistochemistry with anti-myeloperoxidase (MPO) antibody. MPO could be detected after 48 h in the lateral wall and the organ of Corti. After injection of LPS, the threshold of the compound action potential worsened significantly at 48 h in the LPS group. These results suggest that MPO and reactive oxygen species are involved in cochlea dysfunction under inflammatory conditions.

Acute Disease↗
PubMed2001

Apoptosis in the hydropic cochlea of guinea pigs following immune reaction of the endolymphatic sac: immunohistochemical analysis.

Apoptotic change in the cochlea was studied by immunohistochemistry after the injection of keyhole limpet haemocyanin (KLH) into the right endolymphatic sac of guinea pigs. Apoptosis was examined with the specific antibody to single-stranded DNA (ssDNA). Endolymphatic hydrops became evident in the cochlea 1 day after the injection of KLH (n = 6). Increased ssDNA expression could be detected in the spiral ligament and the stria vascularis. The temporal bones in the control group did not show any ssDNA immunoreactivity (n = 6). Apoptosis is the process of the cell death. Our findings imply that apoptotic changes are involved in endolymphatic hydrops. These phenomena could lead to cochlear dysfunction as seen in endolymphatic hydrops.

Animals↗
PubMed2001

Differential effect of Fyn tyrosine kinase deletion on offensive and defensive aggression.

Fyn tyrosine kinase is highly expressed in the limbic system and mice lacking Fyn tyrosine kinase showed increased fearfulness in a variety of tests for anxiety-related behaviors. To investigate the possible role of Fyn tyrosine kinase in aggression, we assessed the aggressive behaviors of the mice lacking the Fyn tyrosine kinase using the resident-intruder and restraint-induced target biting paradigms. The percentage of Fyn-deficient mice that attacked an inanimate target in a restraint tube was higher than that of the control mice. On the contrary, in the resident-intruder paradigm, the percentage of Fyn-deficient mice that attacked the intruder was lower and the Fyn-deficient mice showed a longer latency to attack an intruder. These results suggest a distinct role of Fyn tyrosine kinase in enhancing the offensive aggression and decreasing the defensive aggression. A possible influence of anxiety-phenotype of the Fyn-deficient mice on their abnormal aggressive behavior was discussed.

Aggression↗