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Biomedical subjects

Hideaki Tanaka

Publications and source records attributed to Hideaki Tanaka.

At least 55 records · Page 3Linked to original sources
PubMed2003

Ephrin-A2 regulates position-specific cell affinity and is involved in cartilage morphogenesis in the chick limb bud.

In the developing limb bud, mesenchymal cells show position-specific affinity, suggesting that the positional identity of the cells is represented as their surface properties. Since the affinity is regulated by glycosylphosphatidylinositol (GPI)-anchored cell surface proteins, and by EphA4 receptor tyrosine kinase, we hypothesized that the GPI-anchored ligand, the ephrin-A family, also contributes to the affinity. Here, we describe the role of ephrin-A2 in the chick limb bud. Ephrin-A2 protein is uniformly distributed in the limb bud during early limb development. As the limb bud grows, expression of ephrin-A2 is strong in its proximal-to-intermediate regions, but weak distally. The position-dependent expression is maintained in vitro, and is regulated by FGF protein, which is produced in the apical ectodermal ridge. To investigate the role of ephrin-A2 in affinity and in cartilage morphogenesis of limb mesenchyme, we ectopically expressed ephrin-A2 in the limb bud using the retrovirus vector, RCAS. Overexpressed ephrin-A2 modulated the affinity of the mesenchymal cells that differentiate into autopod elements. It also caused malformation of the autopod skeleton and interfered with cartilage nodule formation in vitro without inhibiting chondrogenesis. These results suggest that ephrin-A2 regulates the position-specific affinity of limb mesenchyme and is involved in cartilage pattern formation in the limb.

Animals↗
PubMed2003

Crystallization and preliminary crystallographic analysis of the importin-beta-SREBP-2 complex.

The nuclear-transport protein importin-beta mediates the nuclear import of the transcription factor SREBP-2 without requiring adaptor proteins such as importin-alpha. An importin-beta-SREBP-2 HLHZ domain complex was purified and crystallized. The crystals belong to space group P2(1)2(1)2(1) and show diffraction to at least 3.0 A resolution. The unit-cell parameters are a = 101.0, b = 113.2, c = 240.0 A. Structure determination using the MAD or SAD method is under way.

Animals↗
PubMed2003

Standardization of ischemic hepatic failure in pigs as a model for bioartificial liver assessment.

PURPOSE: A standard protocol of ischemic liver failure in pigs was examined to establish a system for assessing the efficacy of a bioartificial liver, based on clinical practice. METHODS: The portal blood flow was extracorporeally bypassed into the cervical jugular vein, using a centrifugal blood pump. The portal vein and hepatic artery were then ligated. RESULTS: The maintenance protocol was established as follows: (1) the concentration of the inhaled anesthetic was decreased by 0.2% when the systolic blood pressure was <100 mmHg; (2) the volume of an infusion containing 5% glucose was increased to 10 ml/kg per hour when central venous pressure was <5 mmHg; (3) 20 ml of 50% glucose was injected intravenously when the blood glucose was <50 mg/dl; (4) 2000 units of heparin was injected intravenously when the activated clotting time was <150 s; (5) sodium bicarbonate was given when the blood pH was <7.3; (6) tidal volume was increased by 1 ml/kg when the pCO(2) was >80 mmHg; (7) oxygen was increased by 25% when the pO(2) was <100 mmHg. No vasopressors were used in the experiment. CONCLUSION: Our protocol reduced the operating time and minimized the risk of data deviation that can arise from variations in operating techniques and individual animal conditions. This experimental model is also easy to use as a bridge to transplantation.

Animals↗
PubMed2003

Successful transplantation of a 20% partial liver graft in rats: a technical innovation.

BACKGROUND: Graft-size disparity negatively impacts the postoperative outcome in the setting of adult-to-adult living donor liver transplantation. MATERIALS AND METHODS: As a model for partial liver transplantation in rats using a minimized graft volume, we orthotopically transplanted a 20% reduced-size liver with the two-cuff method. Blood chemistry, graft weight restoration, and labeling indices of 5-bromo-2'-deoxyuridine were estimated. RESULTS: We achieved 100% 2-week survival. Hepatic mass was restored rapidly with early normalization of biochemical parameters, although albumin synthesis capacity required more time to recover. CONCLUSION: This technique has great potential for research as well as a therapeutic approach to regeneration and functional recovery of small liver grafts under various posttransplant circumstances.

Animals↗
PubMed2003

Different hepatitis C virus dynamics of free-virions and immune-complexes after initiation of interferon-alpha in patients with chronic hepatitis C.

BACKGROUND/AIMS: To elucidate the mechanisms of action of interferon (IFN) against hepatitis C virus (HCV), we studied the serum HCV dynamics of free-virions (FV) and immune-complexes (IC) in patients treated with IFN. METHODS: FV and IC were separated by immunoprecipitation using anti-human immunoglobulin and quantified serially using real-time detection-polymerase chain reaction. RESULTS: Initially [1st phase (0-24 h)], the FV decreased more rapidly compared to IC [exponential decay slope (EDS)=1.78+/-0.42 vs. 0.99+/-0.31 log10/day, P<0.001; half-life=5.65+/-2.02 vs. 12.5+/-2.83 h, P<0.0001], but at the 2nd phase (1-14 days), half-life of FV was significantly longer than that of IC (101+/-117 vs. 14.2+/-1.08 h, P<0.005). Regarding response to IFN, the decline slope was not significantly different at the 1st phase, but at the 2nd phase, the FV-HCV RNA decreased more slowly in non-responders than in sustained responders to IFN (EDS=0.05+/-0.02 vs. 0.34+/-0.19 log10/day, P<0.005; half-life=186+/-112 vs. 15.3+/-1.85 h, P<0.005). CONCLUSIONS: The presence of escape mutants from the neutralizing antibodies may be involved in resistance to IFN. Analyzes of FV- and IC-HCV dynamics are useful for predicting the IFN efficacy and understanding the mechanism of IFN action in chronic hepatitis patients.

Adult↗
PubMed2003

Equarin, a novel soluble molecule expressed with polarity at chick embryonic lens equator, is involved in eye formation.

The lens plays an important role in eye development. To investigate the molecular mechanisms involved, we used signal sequence trap screens with a chicken lens cDNA library and identified a novel secreted molecule, equarin. Equarin encodes consensus repeat domains conserved in human SRPX and mouse Urb. In the embryonic eye, equarin transcript is detected exclusively in the lens, and persists in the lens equatorial region in a high-dorsal-to-low-ventral gradient. In vitro analysis of equarin protein indicated that after translation, it is modified, cleaved, and secreted to extracellular locations. Microinjection of equarin mRNA into Xenopus embryos induced abnormal eye development. These data suggest that equarin is involved in eye formation.

Alternative Splicing↗
PubMed2003

Cdk5/p35 and Rho-kinase mediate ephrin-A5-induced signaling in retinal ganglion cells.

Ephrin-As are repulsive axonal guidance cues that regulate retinotectal projection. EphA tyrosine kinases, which are the receptors of ephrin-As, activate signaling cascades leading to cytosckeleton reorganization. Here, we address the role of cyclin-dependent kinase (Cdk) 5 in Eph receptor signaling induced by ephrin-A5. Ephrin-A5 induced a cell morphological response in PC-3M cells that endogenously express Cdk5 and EphA2, a receptor for ephrin-A5. This response was augmented by the transfection of p35, which is a neuronal regulator of Cdk5. While the morphological response of native PC-3M cells was not affected by olomoucine, an inhibitor of Cdk, the response was inhibited in the p35-transfected cells. In retinal ganglion cells, either olomoucine at 20 microM or Y-27632 at 10 microM, an inhibitor of Rho-kinase/ROKalpha/ROCKII, showed maximum inhibitory effect against ephrin-A5 (10 microg/ml)-induced growth cone collapse. Combined application of olomoucine and Y-27632 further suppressed the ephrin-A5-induced response. Ephrin-A5 evoked phosphorylation of Cdk5 at Tyr15 and tau, a substrate of Cdk5 in retinal growth cones. Recombinant herpes simplex virus expressing Cdk5 mutant (kinase-negative or Tyr15 to Ala) showed a dominant-negative effect on the ephrin-A5-induced growth cone collapse. These findings demonstrate that both Cdk5 and the Rho kinase pathway independently contribute to the downstream of ephrin-A-induced signaling in retinal ganglion cells.

Animals↗
PubMed2003

Reduced brain electric activities of frontal lobe in cortical cerebellar atrophy.

OBJECTIVE: To assess the relationship between cerebellum and brain cortical activity without motor factors, we recorded the mid-latency auditory evoked responses (MLRs) with simultaneous recording of the electroencephalography (EEG) at rest in patients with 'pure cortical cerebellar atrophy (CCA)'. METHODS: We studied 12 normal control subjects and non-demented 'pure CCA' patients determined by quantitative magnetic resonance imaging analysis. A comprehensive neuropsychological test battery assessed intelligence, frontal lobe function and word fluency. Spontaneous eyes-closed resting EEG and MLRs were recorded from 20 scalp electrodes and analysed with low-resolution electromagnetic tomography (LORETA) to compute the 3-dimensional intracerebral distribution of electric activity. RESULTS: Neuropsychological tests revealed no differences between CCA and the control. Analysis of EEG and MLRs using classical methods also did not reveal any differences. LORETA analysis indicated significant decrease of alpha2 activity in the left inferior frontal gyrus in CCA. On MLRs, the most significant difference was observed at P1 component, and CCA patients showed significant decrease at the right superior frontal gyrus. CONCLUSIONS: Our results indicated that the frontal lobe and ascending reticular activating system are inhibited in CCA patients, and suggested the involvement of the cerebellum in cortical electric activities irrespective of motor adjunct. SIGNIFICANCE: Quantitative EEG and MLR measurements with LORETA pointed out frontal lobe hypoactivities in pure CCA patients.

Aged↗
PubMed2003

Deletion of a chaperonin 60 beta gene leads to cell death in the Arabidopsis lesion initiation 1 mutant.

Lesion mimic mutants develop spontaneous cell death without pathogen attack. Some of the genes defined by these mutations may function as regulators of cell death, whereas others may perturb cellular metabolism in a way that leads to cell death. To understand the molecular mechanism of cell death in lesion mimic mutants, we isolated a lesion initiation 1 (len1) mutant by a T-DNA tagging method. The len1 mutant develops lesions on its leaves and expresses systemic acquired resistance (SAR). LEN1 was identified to encode a chloroplast chaperonin 60 beta (Cpn60 beta), a homologue of bacterial GroEL. The recombinant LEN1 had molecular chaperone activity for suppressing protein aggregation in vitro. Moreover, len1 plants develop accelerated cell death to heat shock stress in comparison with wild-type plants. The chlorophyll a/b binding protein (CAB) was present in len1 plants at a lower level than in the wild-type plants. These results indicate that LEN1 functions as a molecular chaperone in chloroplasts and its deletion leads to cell death in Arabidopsis.

Adaptation, Physiological↗
PubMed2003

Cognitive dysfunction in cortical cerebellar atrophy correlates with impairment of the inhibitory system.

The aim of the present study was to evaluate the profile of cognitive impairment in patients with cortical cerebellar atrophy (CCA) by measurement of event-related potentials (ERP) and neuropsychological tests. We studied 13 CCA patients and 13 age-, sex- and education-matched normal controls. For ERP recording, we used the conventional auditory oddball task as well as the continuous performance task, which evaluates the attentional performance and ability to control a motor response, i.e., to execute ("Go") or inhibit a motor reaction ("No Go"). Brain electric activity was recorded using 20 scalp electrodes and computed into series of potential distribution maps. For components of ERP, reference-independent measures [global field power (GFP)] were determined, and low-resolution brain electromagnetic tomography (LORETA) was used to compute the three-dimensional intracerebral distribution of electric activity of the P3 component of Go and No Go responses. A comprehensive neuropsychological test battery was also assessed. GFP peak latency was prolonged and GFP peak was attenuated under the No Go condition in patients with CCA, although there were no differences in the auditory oddball task and in the Go condition between the two groups. LORETA showed low activation of frontal source in CCA patients in No Go P3 compared with the controls. However, neuropsychological tests revealed no differences between the two groups. Our results indicate that degeneration of the cerebellum contributes to frontal dysfunction, and suggest this dysfunction is characterized by an impairment of the inhibitory system.

Aged↗
PubMed2003

[Quinacrine administration to a patient with Creutzfeldt-Jakob disease who received a cadaveric dura mater graft--an EEG evaluation].

An old antimalarial drug quinacrine is one of the compounds that can inhibit the accumulation of pathogenic prion protein in cultured neuroblastoma cells. Here we report the result of a clinical trial of quinacrine administration to a patient with Creutzfeldt-Jakob disease (CJD). A 37-year-old woman was admitted to our hospital for ataxic gait, hallucination and dementia in August 2001. As a past history she had received an operation for cerebellar astrocytoma and had a cadaveric dura mater graft at 14 years old. After hospitalization she showed signs of anxiety, stimulus-sensitive myoclonus, decorticate rigidity, then deteriorated pursuit eye movement and voluntary limb movement. The ethics committee of Dokkyo University hospital approved this treatment, and the family of the subject was fully informed as to the nature and purpose of the treatment and gave their consent. One week after the administration of 300 mg/day of quinacrine, voluntary limb movement and pursuit eye movement appeared. There was PSD on EEG, but one week after administration of the quinacrine, PSD disappeared, and a theta wave was recognized as the basic activity on the EEG. But her symptoms deteriorated again after 3 weeks, and PSD appeared. Liver dysfunction developed 2 months after administration, and the administration of quinacrine had to be stopped. Quinacrine was administrated to a CJD patient who received a cadaveric dura mater graft and developed CJD 23 years later. The symptoms of the patient were transiently improved with the disappearance of PSD on EEG. Although the mechanism of quinacrine for the central nervous system is still unclear, this case suggests that quinacrine may be clinically one possible drug for treating CJD.

Adult↗
PubMed2002

Identification and characterization of a soluble cadherin-7 isoform produced by alternative splicing.

We identified an alternative mRNA encoding a novel cadherin-7 isoform by reverse transcriptase-PCR of RNA from day 12 chicken embryos. The alternative mRNA contains 49 bases of insertion in the premembrane region, leading to the substitution of 14 amino acids and the introduction of a premature stop codon. Identification of a 49-bp insertion sequence in the genomic DNA corresponding to the intron of the cadherin-7 gene suggests that alternative splicing is the cause of the alternative mRNA. Transient expression of the variant form in COS-7 or 293 cells produced a soluble protein. Aggregation assays and immunoprecipitation showed that the variant protein interacts with full-length cadherin-7 in vitro and in vivo and inhibits full-length cadherin-7-mediated cell adhesion. Immunohistochemistry revealed that the variant form was strongly expressed in dermomyotomes rather than in migrating neural crest cells, in contrast to the full-length cadherin-7, suggesting differential regulation of splicing and possible roles of variant cadherin-7 in the development of dermomyotomes and other tissues.

Alternative Splicing↗
PubMed2002

Ephrin-A5 restricts topographically specific arborization in the chick retinotectal projection in vivo.

The retinotectal map is the best characterized model system to study how axons respond to guidance cues during the formation of the nervous system. Recent studies have shown that the critical event in forming this map is topographic-specific axon branching. To elucidate the in vivo role of the repulsive cue ephrin-A5 in this event, we used chromophore-assisted laser inactivation (CALI) to generate acute loss of ephrin-A5 function in localized areas of the posterior tectum of chick embryos in ovo and analyzed the resulting changes of retinal projections during initial outgrowth (E11) and when retinal axons arborize in the deep layers in the tectum (E12). We confirmed that ephrin-A5 functions to restrict initial axon outgrowth at E11. At E12, CALI of ephrin-A5 did not affect the extent of axon outgrowth on the tectal surface but instead caused ectopic arborization posterior to the topographically correct site in deeper layers of the tectum. This shows that ephrin-A5 restricts arborization during this critical process for developing the retinotopic map. CALI provides an approach to inactivate in vivo function in higher vertebrates with high temporal and spatial specificity that may have wide application.

Animals↗
PubMed2002

Structural analysis of chick ephrin-A2 by function-blocking and non-blocking monoclonal antibodies.

Ephrins, ligands for the Eph family of receptor tyrosine kinases, play key roles in diverse biological processes. In this study, we determined the epitopes and kinetic parameters of function-blocking (B3) and non-blocking (IV) monoclonal antibodies (mAbs) recognizing chick ephrin-A2. We show that the epitope for the non-blocking mAb is the residue Asp(105) of chick ephrin-A2. However, the binding of the function-blocking mAb depends mostly on residue Ser(108) and its epitope may reside within residues 105-132, which appear crucial for the receptor interaction site. Kinetic studies suggest a possible mechanism why mAb IV, despite recognizing a region very close to the mAb B3 epitope, fails to block the ligand-receptor interaction.

Animals↗
PubMed2002

Insufficient processing resources in Parkinson's disease: evaluation using multimodal event-related potentials paradigm.

The purpose of our study was to demonstrate impaired allocation of processing resources in non-demented patients with early-stage mild Parkinson's disease (PD) using a multimodal event-related potential (ERP) paradigm. The multimodal ERP paradigm was performed in 18 non-demented medicated patients with early-stage PD (Mini-Mental State Examination Score >26) and 16 matched normal controls, the Global Field Power (GFP) was employed for ERP components analysis, and the new modified Wisconsin Card Sorting Test (WCST) was used to evaluate frontal lobe function. Patients with PD did not exhibit novelty P3s, and P3 latency to non-target novel stimuli in visual and auditory modalities was significantly longer in PD patients than in controls. P3 amplitude for the target stimuli (P3b) was higher in PD in both auditory and visual modalities; however, P3b latency was not different between the two groups. Patients with PD showed a significantly lower score of achieved categories and made more perseverative errors in WCST as compared to controls. Our results showed that there were no natural novelty P3s in patients with PD; this finding suggests that non-demented patients with mild PD do not have sufficient mental resources to allocate to the central executive, due to dysfunction of the frontal lobe.

Acoustic Stimulation↗
PubMed2002

Ephrin-B3-EphA4 interactions regulate the growth of specific thalamocortical axon populations in vitro.

The role was studied of ephrin-B3, a ligand of the Eph family of tyrosine kinase receptors, in the formation of cortical connectivity. In situ hybridization and immunohistochemistry showed that EphA4, a receptor of ephrin-B3, was expressed in the lateral thalamus (visual and somaotosensory thalamus) of the developing rat brain, but not in the medial thalamic nuclei which project to the limbic cortex. Correspondingly, ephrin-B3 was expressed strongly in the developing limbic cortex including amygdala, entorhinal cortex and hippocampus. To examine the action of ephrin-B3 on thalamic axons, either lateral or medial thalamic explants were cultured on membranes obtained from ephrin-B3-expressing COS cells. Axonal growth was inhibited for cells from the lateral thalamus but not from the medial thalamus. These results suggest that ephrin-B3 contributes to regional specificity by suppressing axonal growth of lateral thalamic neurons.

Animals↗