Biomedical subjects

H Itoh

Publications and source records attributed to H Itoh.

At least 181 records · Page 10Linked to original sources

PubMed2000

Effects of geometry of hydroxyapatite as a cell substratum in BMP-induced ectopic bone formation.

Three different types of porous hydroxyapatite with pore sizes of 100-200 micrometer in diameter-porous particles of hydroxyapatite (PPHAP), porous blocks of hydroxyapatite (PBHAP), and honeycomb-shaped hydroxyapatite (HCHAP)-were compared in terms of their abilities to induce osteogenesis when implanted subcutaneously with recombinant human BMP-2 into rats and extracted at 1, 2, 3, and 4 weeks. Histologically, direct bone formation occurred in PPHAP and PBHAP while only endochondral ossification took place in HCHAP. Interestingly, cartilage in the central zones and bone in the orifice zones of the tunnels of the HCHAP were observed at 2 weeks. After 3 weeks, the cartilage disappeared and bone formation occurred throughout the inner surface of the tunnels of the HCHAP, always leaving space for capillaries within the tunnels. Alkaline phosphatase activity and osteocalcin content were the highest in HCHAP among the three hydroxyapatite implants. These results clearly indicate that BMP-induced bone formation is highly dependent on the geometry of the carrier, which provides feasible structural factors for vascularization.

Alkaline Phosphatase↗

PubMed2000

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

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

Brain Mapping↗

PubMed2000

Synthesis and receptor sites of endothelin-1 in the rat liver vasculature.

Immunocytochemical localization of big endothelin-1 (big ET-1), ET-1, and ET receptor A and B (ET(A) and ET(B)), and gene expression of prepro ET-1 mRNA were examined on the rat liver vasculature. Immunoreactivities for big ET-1 and ET-1 were preferentially seen along the endothelium of interlobular veins (IV) and artery (IA), although the staining intensity was more pronounced in IV. Expression of preproET-1 mRNA was detected in both vascular endothelia and the signal intensity was more prevalent in IV. Immunoelectron microscopy showed that rough endoplasmic cisterns were immunoreactive for big ET-1, while Weibel-Palade (WP) bodies, a storage site for ET-1, were immunoreactive for ET-1 in endothelial cells of IV. These results indicate that endothelial cells of IV are the major site of synthesis of ET-1, which is extracellularly secreted by degranulation and/or exocytosis of WP bodies. Hepatic stellate cells (HSCs), especially of the plasma membrane of perisinusoidal and interhepatocellular processes, were immunoreactive for both ET(A) and ET(B) receptor antibodies. These findings suggest that ET-1 receptor-mediated HSC contraction is involved in the regulation of hepatic sinusoidal blood flow as previously cited in mammalian liver cirrhosis. We also showed that sarcolemma and caveoles in the smooth muscle cells of the media of IV, and its branches before reaching the hepatic sinusoids, were immunoreactive for ET(A) receptor antibody. The results suggest that such vessels, which contains a large amount of hepatic blood inflow, participate in pump mechanism toward hepatic sinusoidal circulation in a receptor-mediated paracrine fashion.

Animals↗

PubMed2000

A milestone for normal development of the infantile brain detected by functional MRI.

OBJECTIVE: To investigate the relationship between cerebral cortical function and white matter myelination in the visual pathway in the evaluation of normal brain development. METHODS: The authors performed quantitative analysis of white matter myelination detected with conventional T1-weighted spin echo (SE) MRI and brain functional MRI (fMRI) using echoplanar imaging with photic stimulation in 27 neurologically normal infants (age range, 0 to 22 weeks). RESULTS: An age-dependent gradual increase in signal intensity was observed in optic radiation on the T1-weighted SE images, indicating progression of white matter myelination. A rapid age-dependent reverse in signal response was observed on fMRI. Infants older than 8 weeks showed a stimulus-induced signal decrease in the visual cortex, whereas infants younger than 7 weeks showed a signal increase. CONCLUSIONS: A rapid inversion of response revealed by fMRI with photic stimulation in infants suggests a change in oxygen consumption during neuronal activation, which is related to rapid synapse formation and accompanying increased metabolism. fMRI can detect dynamic metabolic changes during brain maturation, which is a different developmental process from white matter myelination. The metabolic changes detected by fMRI provide a milestone for the evaluation of normal brain development.

Brain↗

PubMed2000

Functional interaction of the immunosuppressant mizoribine with the 14-3-3 protein.

Mizoribine (MIZ) is a novel imidazole nucleoside with immunosuppressive activity. MIZ has been approved in Japan and combination therapy with MIZ and glucocorticoids has been used after renal transplantation and for lupus nephritis and rheumatoid arthritis. In this study, we identify 14-3-3 proteins as MIZ-binding proteins. 14-3-3 proteins interact with many proteins involved in cellular signaling, including the glucocorticoid receptor (GR). The 14-3-3/GR interaction enhances the transcriptional activity of the receptor. We show that MIZ affects the conformation of 14-3-3 proteins and enhances the interaction of GR and 14-3-3eta dose dependently in vitro. MIZ also has a stimulatory effect on transcriptional activation by the GR. Our results point to the possibility that one mechanism for the therapeutic effect of MIZ could be to regulate the GR function via 14-3-3 proteins.

14-3-3 Proteins↗

PubMed2000

NESK, a member of the germinal center kinase family that activates the c-Jun N-terminal kinase pathway and is expressed during the late stages of embryogenesis.

The c-Jun N-terminal kinase (JNK) signaling pathway plays a crucial role in cellular responses stimulated by stress-inducing agents and proinflammatory cytokines. The group I germinal center kinase family members selectively activate the JNK pathway. In this study, we have isolated a mouse cDNA encoding a protein kinase homologous to Nck-interacting kinase (NIK), a member of the group I germinal center kinase family. This protein kinase is expressed during the late stages of embryogenesis, but not in adult tissues, and thus named NESK (NIK-like embryo-specific kinase). NESK selectively activated the JNK pathway when overexpressed in HEK 293 cells but did not stimulate the p38 kinase or extracellular signal-regulated kinase (ERK) pathways. NESK-induced JNK activation was inhibited by the dominant negative mutants of MEKK1 and MKK4. Tumor necrosis factor (TNF)-alpha or TNF receptor-associated factor 2 (TRAF2) stimulated the NESK activity. Furthermore, the dominant negative NESK mutant inhibited the JNK activation induced by TNF-alpha or TRAF2. These results suggest that NESK, a novel activator of the JNK pathway, functions in coupling TRAF2 to the MEKK1 --> MKK4 --> JNK kinase cascade during the late stages of mammalian embryogenesis.

Amino Acid Sequence↗

PubMed2000

Identification and characterization of negative regulatory elements of the human telomerase catalytic subunit (hTERT) gene promoter: possible role of MZF-2 in transcriptional repression of hTERT.

Human telomerase reverse transcriptase (hTERT) is a catalytic subunit of human telomerase and is a critical determinant of the enzymatic activity of telomerase. Expression of hTERT is known to be regulated mainly at the transcriptional level. In the present study, using transient expression assays, we identified a 400 bp silencer of the hTERT promoter between -776 and -378 upstream of the proximal core promoter. The inhibitory effects of this silencer were enhanced with cellular differentiation. A computer-assisted homology search identified multiple binding motifs for myeloid-specific zinc finger protein 2 (MZF-2) within this region. Mutation introduced in these sites resulted in significant activation of hTERT transcription. Gel shift assays demonstrated that MZF-2 proteins specifically bound to these sites. Overexpression of MZF-2 in cells led to down-regulation of hTERT transcription as well as telomerase activity. These findings suggest that the 400 bp region upstream of the hTERT core promoter that we identified functions as a negative regulatory region and that MZF-2 may be an effector of negative regulation of hTERT.

Base Sequence↗

PubMed2000

Stepping rotation of F1-ATPase visualized through angle-resolved single-fluorophore imaging.

Orientation dependence of single-fluorophore intensity was exploited in order to videotape conformational changes in a protein machine in real time. The fluorophore Cy3 attached to the central subunit of F(1)-ATPase revealed that the subunit rotates in the molecule in discrete 120 degrees steps and that each step is driven by the hydrolysis of one ATP molecule. These results, unlike those from the previous study under a frictional load, show that the 120 degrees stepping is a genuine property of this molecular motor. The data also show that the rate of ATP binding is insensitive to the load exerted on the rotor subunit.

Adenosine Triphosphate↗

PubMed2000

Inhibition of rho-associated kinase results in suppression of neointimal formation of balloon-injured arteries.

BACKGROUND: Rho-associated kinase (ROCK), an effector of small GTPase Rho, regulates vascular tone via a calcium sensitization mechanism and plays a key role in the pathogenesis of hypertension. However, its role in vascular growth remains unclear. METHODS AND RESULTS: Y-27632, a specific ROCK inhibitor, and the overexpression of dominant-negative ROCK suppressed the mitogen-induced DNA synthesis of cultured vascular smooth muscle cells (VSMCs), which indicates the essential role of ROCK in the control of VSMC proliferation in vitro. Y-27632 also suppressed the chemotaxis of VSMCs. Male Wistar rats were systemically given Y-27632 (35 to 70 mg. kg(-1). day(-1)) through an intraperitoneal infusion. The neointimal formation of balloon-injured carotid arteries was significantly suppressed in Y-27632-treated rats (intima/media ratio, 0.22+/-0.02) compared with vehicle-treated rats (intima/media ratio, 0.92+/-0.21) or hydralazine-treated rats with a similar blood pressure decrease (intima/media ratio, 1.03+/-0.15). The phosphorylation of myosin phosphatase and myosin light chain was elevated in injured arteries in a Y-27632-sensitive manner, indicating the augmentation of ROCK activity in neointimal formation. The downregulation of the cyclin-dependent kinase inhibitor p27(kip1) in injured vessels was reversed by Y-27632 treatment, reflecting the antiproliferative effect of ROCK inhibition in vivo. CONCLUSIONS: We conclude that ROCK plays a key role in the process of neointimal formation after balloon injury. Thus, the inhibition of ROCK may be a potential therapeutic strategy for treating vascular proliferative disorders and hypertension.

Amides↗

PubMed2000

Thrombin-induced inhibition of myoblast differentiation is mediated by Gbetagamma.

Thrombin has been shown to inhibit skeletal muscle differentiation. However, the mechanisms by which thrombin represses myogenesis remain unknown. Since the thrombin receptor couples to G(i), G(q/11) and G(12), we examined which subunits of heterotrimeric guanine nucleotide-binding regulatory proteins (Galpha(i), Galpha(q/11), Galpha(12) or Gbetagamma) participate in the thrombin-induced inhibition of C2C12 myoblast differentiation. Galpha(i2) and Galpha(11) had no inhibitory effect on the myogenic differentiation. Galpha(12) prevented only myoblast fusion, whereas Gbetagamma inhibited both the induction of skeletal muscle-specific markers and the myotube formation. In addition, the thrombin-induced reduction of creatine kinase activity was blocked by the C-terminal peptide of beta-adrenergic receptor kinase, which is known to sequester free Gbetagamma. These results suggest that the thrombin-induced inhibition of muscle differentiation is mainly mediated by Gbetagamma.

Animals↗

PubMed2000

Mouse hepatocyte growth factor activator gene: its expression not only in the liver but also in the gastrointestinal tract.

A cDNA encoding mouse hepatocyte growth factor activator (HGFA) has been cloned by RT-PCR, based on the screening result from the database of expressed sequence tags. Subsequently, its gene was cloned from a mouse genomic bacterial artificial chromosome library using the cDNA as a probe. Sequencing analysis revealed that mouse HGFA protein deduced from the cDNA, similar to its human and rat counterparts, has two epidermal growth factor-like domains, type 1 and 2 fibronectin homology domains, a single kringle domain and a catalytic domain of serine proteinase, and the gene consists of 14 exon spanning approximately 7.5 kb. Interestingly, mouse HGFA mRNA was detected not only in the liver but also in the gastrointestinal tract by RNA blot analysis. Since hepatocyte growth factor (HGF) is up-regulated in the damaged gastrointestinal mucosa, our present data suggest that HGFA might activate proHGF directly in the gastrointestinal mucosa and play an important role in wound repair throughout the gastrointestinal tract.

Amino Acid Sequence↗

PubMed2000

Cardiac fibrosis in mice lacking brain natriuretic peptide.

Cardiac fibrosis, defined as a proliferation of interstitial fibroblasts and biosynthesis of extracellular matrix components in the ventricles of the heart, is a consequence of remodeling processes initiated by pathologic events associated with a variety of cardiovascular disorders, which leads to abnormal myocardial stiffness and, ultimately, ventricular dysfunction. Brain natriuretic peptide (BNP) is a cardiac hormone produced primarily by ventricular myocytes, and its plasma concentrations are markedly elevated in patients with congestive heart failure and acute myocardial infarction. However, its precise functional significance has been undefined. In this paper, we report the generation of mice with targeted disruption of BNP (Nppb(-/-) mice). We observed multifocal fibrotic lesions in the ventricles from Nppb(-/-) mice. No signs of systemic hypertension and ventricular hypertrophy are noted in Nppb(-/-) mice. In response to ventricular pressure overload, focal fibrotic lesions are increased in size and number in Nppb(-/-) mice, whereas no focal fibrotic changes are found in wild-type littermates (Nppb(+/+) mice). This study establishes BNP as a cardiomyocyte-derived antifibrotic factor in vivo and provides evidence for its role as a local regulator of ventricular remodeling.

Animals↗

PubMed2000

A new system for the anterior restoration and fixation of thoracic spinal deformities using an endoscopic approach.

STUDY DESIGN: A report on the results of animal experiments in which the authors' new system that enables the anterior correction and fixation of thoracic spinal deformity by use of a thoracic endoscope and small incisions was used. The results suggest that the new approach involving thoracoscopic anterior correction and fusion for thoracic spinal deformity could be adapted successfully for the clinic. OBJECTIVE: To develop a new system that enables the anterior correction and fixation of thoracic spinal deformity by use of a thoracic endoscope and small incisions. SUMMARY OF BACKGROUND DATA: Anterior correction and fusion through open thoracotomy have been applied mainly for thoracic scoliosis because this approach provides effective correction with short fusion. However, excellent visualization of the spine during thoracic surgical procedures led to the development of thoracoscopic spine surgery. Therefore, the authors postulated that a thoracoscopic approach could allow not only discectomy but also correction and fusion of the deformed thoracic spine in a single surgical event. METHODS: The vertebral columns and attached thoracic walls were dissected from fresh porcine cadavers and used in the experimental surgery to evaluate the new system. As a next step, thoracoscopic surgery using this new system was performed on four or five vertebrae of five live pigs with an average weight of 50 kg. Initially, the discs and rib heads were removed through the thoracic wall. For each vertebra, a screw connected to a shaft used as a screw holder was inserted through the thoracic wall in a posterior to anterior direction. Each shaft then was linked outside the body to an outrigger. This outrigger was used to both create and restore scoliosis. A rod then was introduced through the thoracic wall and fixed to the screw heads. RESULTS: The animal experiments clearly showed that it was possible to change the Cobb angles of the spine through the use of the outrigger apparatus. In cadaver experiments, it was possible to create scoliosis and re-store it by 25 degrees to 35 degrees. Also, surgery on live pigs resulted in scoliosis of approximately 30 degrees, which means approximately 5 degrees to 10 degrees for each disc space. The procedures used also demonstrated that it was possible to fix a rod, introduced into the pleural cavity through a port, with screw heads. CONCLUSIONS: The use of this system successfully changed the Cobb angle of the spine, which suggests that its use should make it possible to correct spinal deformities. This apparatus also succeeded in fixing the rods in the screw heads, which raises the possibility of its use in spinal fixation. The authors believe that this apparatus could be adapted successfully for clinical use. Studies currently are under way in clinics using the new system.

Animals↗

PubMed2000

Regulation of the golgi structure by the alpha subunits of heterotrimeric G proteins.

Disassembly of the Golgi apparatus is elicited by the action of nordihydroguaiaretic acid (NDGA) and this disassembly is prevented by the activation of heterotrimeric G proteins. In the present study we showed that overexpression of Galpha(z) or Galpha(i2) significantly suppresses the disassembly of the Golgi apparatus induced by NDGA. Overexpression of Gbeta(1)gamma(2), on the other hand, had no effect on NDGA-induced Golgi disassembly. Galpha(z) neither blocked Golgi disassembly induced by brefeldin A or nocodazole, nor interfered with protein transport, suggesting its specificity on the action of NDGA. Our results suggest that the alpha subunits of heterotrimeric G proteins are responsible for the maintenance of the Golgi structure.

Animals↗

PubMed2000

G(i)-dependent activation of c-Jun N-terminal kinase in human embryonal kidney 293 cells.

Heterotrimeric G proteins stimulate the activities of two stress-activated protein kinases, c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase in mammalian cells. In this study, we examined whether alpha subunits of G(i) family activate JNK using transient expression system in human embryonal kidney 293 cells. Constitutively activated mutants of Galpha(i1), Galpha(i2), and Galpha(i3) increased JNK activity. In contrast, constitutively activated Galpha(o) and Galpha(z) mutants did not stimulate JNK activity. To examine the mechanism of JNK activation by Galpha(i), kinase-deficient mutants of mitogen-activated protein kinase kinase 4 (MKK4) and 7 (MKK7), which are known to be JNK activators, were transfected into the cells. However, Galpha(i)-induced JNK activation was not blocked effectively by kinase-deficient MKK4 and MKK7. In addition, activated Galpha(i) mutant failed to stimulate MKK4 and MKK7 activities. Furthermore, JNK activation by Galpha(i) was inhibited by dominant-negative Rho and Cdc42 and tyrosine kinase inhibitors, but not dominant-negative Rac and phosphatidylinositol 3-kinase inhibitors. These results indicate that Galpha(i) regulates JNK activity dependent on small GTPases Rho and Cdc42 and on tyrosine kinase but not on MKK4 and MKK7.

Cells, Cultured↗

PubMed2000

Magnetoresistance of ferromagnetic tunnel junctions in the double-exchange model

We conduct a theoretical study of the temperature dependence of the spin polarization ( P) and the magnetoresistance (MR) ratio using the double exchange (DE) model for ferromagnetic tunnel junctions with half-metallic systems. It is shown that the strong exchange coupling in the DE model plays an important role in the temperature dependence of both P and the MR ratio; their values can be less than the maximum values expected for half-metallic systems at low temperatures, and the MR ratio decreases more rapidly than P with increasing temperature. The calculated results, however, indicate that the MR ratio may still be large at high temperatures near the Curie temperature.

Journal Article↗

PubMed2000

Plasma homovanillic acid in the prodromal phase of schizophrenia.

BACKGROUND: Plasma levels of homovanillic acid (pHVA) have been used as a peripheral measure of central dopaminergic activity. Despite a large body of studies investigating pHVA in schizophrenia, little is known about pHVA in patients in the prodromal phase of the illness. METHODS: Plasma HVA levels of 12 male outpatients meeting DSM-III-R criteria for the prodromal phase of schizophrenia at the time of blood sampling (who later developed psychotic symptoms) were compared with those of 12 normal male healthy volunteers. Task amounts in the Kraepelin arithmetic test at the time of blood sampling were compared between the prodromal patients and normal controls and were correlated with pHVA levels. RESULTS: The prodromal patients had significantly higher pHVA levels compared with normal control subjects. The mean amount of the arithmetic task for the prodromal patients was significantly less than that for controls. In the patient group, a significant negative correlation was observed between pHVA levels and the task amounts. CONCLUSIONS: Data from the present study indicate the presence of dopaminergic dysfunction in the prodromal stage of schizophrenia that is associated with neuropsychological impairment. Increased pHVA levels in the prodromal patients may have implications for early detection of schizophrenia.

Acute Disease↗

PubMed2000

[Healing in the intestinal anastomosis--comparison of the Albert-Lembert and Gambee methods].

An experimental comparative histomorphological study was made on intestinal healing processes following an Albert-Lembert suture with approximation of the serosal surface and Gambee's layer to layer anastomosis of a dog. There was no obvious complications such as postoperative hemorrhage, anastomotic stenosis or anastomotic leakage. Although both anastomoses resulted in a good healing process, Gambee's layer to layer suture, which caused the submucosal layers to face each other, showed better wound healing at the anastomosis in terms of layer to layer attachment. As a conclusion, Gambee's layer to layer anastomosis seemed to be a better anastomotic technique in terms of wound healing for the intestinal anastomosis.

Anastomosis, Surgical↗