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Biomedical subjects

M Imai

Publications and source records attributed to M Imai.

At least 91 records · Page 5Linked to original sources
PubMed2000

[Anesthetic management of heart transplantation].

We anesthetized a 47-yr-old man with end-stage hypertrophic cardiomyopathy for heart transplantation. This is the first case of heart transplantation from a patient with brain death, since the organ transplantation law had become valid in Japan. Anesthesia was induced and maintained with fentanyl and diazepam. Aseptic technique was used in inserting and securing all catheters. The patient was assisted by left ventricular assist system, and hemodynamic suppression at anesthetic induction was trivial. Since complete AV block was present at the termination of cardiopulmonary bypass (CPB), VVI pacing and infusion of isoproterenol were started. In addition, nitroglycerin was given for pulmonary vasodilation. The cardiovascular support used for weaning from CPB included dobutamine, isoprote-renol, dopamine and milrinone. Following weaning from CPB sinus rhythm appeared spontaneously and function of the transplanted heart was satisfactory. When the patient was transported to ICU reduction in doses of catecholamines was possible, and dopamine and milrinone were infused. The patient was extubated 10 hours after admission to ICU.

Anesthesia, Inhalation↗
PubMed2000

CD39 modulates endothelial cell activation and apoptosis.

BACKGROUND: CD39 is the dominant vascular nucleoside triphosphate diphosphohydrolase (NTPDase) that exerts major effects on platelet reactivity by the regulated hydrolysis of extracellular adenine nucleotides. The effects of NTPDases on endothelial cell (EC) activation and apoptosis remain unexplored. MATERIAL AND METHODS: Recombinant replication-deficient adenoviruses were constructed with human CD39 cDNA (rAdCD39) or the bacterial beta-galactosidase (rAdbetagal). RESULTS: Intact human umbilical vein EC cultures infected with rAdCD39 had substantial and stable increases in NTPDase biochemical activity (14.50 +/- 3.50 Pi nmole/well/min), when contrasted with noninfected cells (0.95 +/- 0.002) and rAdbetagal infected cells (1.01 +/- 0.02; p<0.005). Increased NTPDase activity efficiently inhibited immediate type 2Y purinergic receptor (P2Y)-mediated EC activation responses viz. von Willebrand factor secretion in response to extracellular ATP. In addition, CD39 up-regulation blocked ATP-induced translocation of the transcription nuclear factor (NF)-kappaB to the cell nucleus, and abrogated transcription of mRNA encoding E-selectin, and consequent protein synthesis. CD39 also decreased the extent of apoptosis triggered by putative type-2X purinergic (P2X7) receptors in response to high concentrations of extracellular ATP in vitro. CONCLUSION: These properties of CD39 indicate primary vascular protective effects with potential therapeutic applications.

Adenosine Triphosphatases↗
PubMed1999

Suppression of ATP diphosphohydrolase/CD39 in human vascular endothelial cells.

Vascular ATP diphosphohydrolase/CD39 is an endothelial cell membrane protein with both ecto-ATPase and ecto-ADPase activities. Suppression of constitutive CD39 expression may result in elevated concentrations of ATP and ADP at the vascular interface that could predispose to thrombosis and inflammation. To study the effects of suppression of CD39 synthesis, stable 25-base antisense chimeric oligonucleotides targeting sequences at the 5' region of CD39 were designed. Transfection of these stable oligomers into cultured human endothelial cells resulted in dramatic decreases in levels of CD39 mRNA transcripts. Following transfection with antisense oligonucleotides, total ADPase activity fell from 26.0 +/- 3.1 in control cultures to 9.5 +/- 3.4 nmol of P(i) min(-1) (mg of protein)(-1) (p < 0.005); suppression of CD39 protein expression was also observed by Western blotting. Decreases in ATP diphosphohydrolase activity were associated with increases in concentrations of extracellular purine nucleotides released following stimulation of endothelial cells. Rates of initial hydrolysis of extracellular ATP released from purinergic agonist-stimulated endothelial cells decreased from 17.9 +/- 5.0 to 4.8 +/- 0.5 pmol min(-1) per 10(6) cells (p < 0.005) in antisense transfected cells. Therefore, CD39 regulates extracellular ATP concentrations and may be an important modulator of purinergic receptor activity in vascular endothelial cells.

Adenosine Diphosphate↗
PubMed1999

Identification of four new members of the rat prolactin/growth hormone gene family.

The rodent prolactin (PRL)/growth hormone (GH) gene family currently consists of at least 14 distinct genes that are expressed mainly in pituitary, uterus, and/or placenta. We report here the identification of novel four members from rat with significant homology to PRL. The encoding proteins are not homologs of other known members of this hormone family. The four new cDNAs were assigned to PRL family based on sequence homology and were referred to as PRL-like protein-I (PLP-I), PLP-J, PLP-K, and PLP-L, following the current naming order of rodent PLP family, where PLP-H is the most recent gene. They encode amino acids with 211-228 amino acids, and 34-38% identity with PRL. All have one or two N-linked glycosylation sites. Among the examined rat tissues by Northern blot analysis, only PLP-I was expressed in testis. Our results indicate that the rodent PRL/GH gene family is large with at least 18 distinct genes.

Amino Acid Sequence↗
PubMed1999

The eukaryotic polypeptide chain releasing factor (eRF3/GSPT) carrying the translation termination signal to the 3'-Poly(A) tail of mRNA. Direct association of erf3/GSPT with polyadenylate-binding protein.

The mammalian GTP-binding protein GSPT, whose carboxyl-terminal sequence is homologous to the eukaryotic elongation factor EF1alpha, binds to the polypeptide chain releasing factor eRF1 to function as eRF3 in the translation termination. The amino-terminal domain of GSPT was, however, not required for the binding. Search for other GSPT-binding proteins in yeast two-hybrid screening system resulted in the identification of a cDNA encoding polyadenylate-binding protein (PABP), whose amino terminus is associating with the poly(A) tail of mRNAs presumably for their stabilization. The interaction appeared to be mediated through the carboxyl-terminal domain of PABP and the amino-terminal region of GSPT. Interestingly, multimerization of PABP with poly(A), which is ascribed to the action of its carboxyl-terminal domain, was completely inhibited by the interaction with the amino-terminal domain of GSPT. These results indicate that GSPT/eRF3 may play important roles not only in the termination of protein synthesis but also in the regulation of mRNA stability. Thus, the present study is the first report showing that GSPT/eRF3 carries the translation termination signal to 3'-poly(A) tail ubiquitously present in eukaryotic mRNAs.

Eukaryotic Cells↗
PubMed1999

Cloning of a stretch-inhibitable nonselective cation channel.

A homologue of the capsaicin receptor-nonselective cation channel was cloned from the rat kidney to investigate a mechanosensitive channel. We found this channel to be inactivated by membrane stretch and have designated it stretch-inactivated channel (SIC). SIC encodes a 563-amino acid protein with putative six transmembrane segments. The cDNA was expressed in mammalian cells, and electophysiological studies were performed. SIC-induced large cation currents were found to be regulated by cell volume, with currents being stimulated by cell shrinkage and inhibited by cell swelling. Single channel analysis showed a conductance of 250 pS with cation permeability (PCl/PNa < 0.1), and the channel possessed some of the characteristics of a stretch-inactivated channel in that it was permeable to calcium, sensitive to membrane stretch, and blocked by Gd3+. Therefore, we cloned one of the mechanosensitive cation channels of mammals, which is considered to regulate Ca2+ influx in response to mechanical stress on the cell membrane.

Amino Acid Sequence↗
PubMed1999

Structural elements and limited proteolysis of CD39 influence ATP diphosphohydrolase activity.

CD39, the mammalian ATP diphosphohydrolase (ATPDase), is thought to contain two transmembrane domains and five "apyrase conserved regions" (ACR) within a large extracellular region. To study the structure of this ectoenzyme, human CD39 was modified by directed mutations within these ACRs or by sequential deletions at both termini. ATPDase activity was well preserved with FLAG tagging, followed by the removal of either of the demonstrated C- or N-transmembrane regions. However, deletions within ACR-1 (aa 54-61) or -4 (aa 212-220), as well as truncation mutants that included ACR-1, -4, or -5 (aa 447-454), resulted in substantive loss of biochemical activity. Intact ACR-1, -4, and -5 within CD39 are therefore required for maintenance of biochemical activity. Native and mutant forms of CD39 lacking TMR were observed to undergo multimerization, associated with the formation of intermolecular disulfide bonds. Limited tryptic cleavage of intact CD39 resulted in two noncovalently membrane-associated fragments (56 and 27 kDa) that substantially augmented ATPDase activity. Glycosylation variation accounted for minor heterogeneity in native and mutant forms of CD39 but did not influence ATPDase function. Enzymatic activity of ATPDase may be influenced by certain posttranslational modifications that are relevant to vascular inflammation.

Adenosine Triphosphatases↗
PubMed1999

Cloning and characterization of 5'-flanking region of mouse non-selective cation channel 1.

We have previously cloned mouse non-selective cation channel 1 (mNSC1) cDNA inducing cation current, from a mouse insulin secreting beta-cell line, MIN6. The current has characteristics of the Ca2+-activated non-selective (CAN) cation channel, and the mRNA is localized in the brain, heart, and lung. To understand the molecular mechanisms of the transcriptional regulation, we have cloned and characterized the 5'-flanking region of mNSC1. By the PCR method, we obtained 987 bp of mouse genomic fragment. The computer program-based analysis revealed that it contained several consensus motifs; insulin responsive element (IRE), AP-2, PEA3, and GC box-like region. But there were neither typical TATA box nor CAAT box. Primer extension analysis and RNase protection assay were performed to identify the transcription start site. Transient transfection analyses using a series of 5'-end deletion and reporter gene constructs with CHO and LA-4 cells demonstrated some relatively active regions. The significantly active border correlated with IRE consensus with CHO cells. This observation may support that CAN current is activated by insulin.

Animals↗
PubMed1999

Expression of an mNSC1 in mammalian cells.

We have cloned a cDNA inducing a cation-permeable current (mNSC1) from pancreatic beta-cells, which shows niflumate-sensitive current in Xenopus oocytes. To elucidate the expression in mammalian cells, mNSC1 was expressed in CHO cells. The reversal potential by mNSC1 was shifted toward positive which was significantly reversed by flufenamic acid. Single-channel analysis showed a characteristic of a Ca-activated nonselective cation channel. Therefore, we may conclude that mNSC1 expresses a fenamates-sensitive cation channel, inducing membrane depolarization in a mammalian cell.

Animals↗
PubMed1999

Plasminogen activator inhibitor 1 in central serous chorioretinopathy.

PURPOSE: To describe the plasma level of plasminogen activator inhibitor 1, the major antifibrinolytic agent, in eyes with central serous chorioretinopathy, in which choroidal thrombosis is suspected as the underlying condition based on the findings of choroidal hyperpermeability in indocyanine green angiograms. METHODS: Plasminogen activator inhibitor 1 concentrations in the plasma of 17 patients with central serous chorioretinopathy were measured by enzyme-linked immunosorbent assay and compared with those in 12 age-matched normal volunteers. RESULTS: The plasma levels of plasminogen activator inhibitor 1 in patients with central serous chorioretinopathy (range, 25 to 439 ng/ml; median, 87 ng/ml) were significantly increased compared with those in normal volunteers (range, 7 to 84 ng/ml; median, 36 ng/ml) (P = .0013, Mann-Whitney U test). CONCLUSION: Increased plasminogen activator inhibitor 1 concentrations in patients with central serous chorioretinopathy support the hypothesis that the choroidal hyperpermeability disclosed by indocyanine green angiography is caused from impaired fibrinolysis and the resulting thrombotic occlusion in the choroidal veins.

Adult↗
PubMed1999

Progression of defects in the central 10-degree visual field of patients with retinitis pigmentosa and choroideremia.

PURPOSE: To determine the progression of defects in the central 10-degree visual field in patients with retinitis pigmentosa and choroideremia by means of univariate linear regression with respect to the mean deviation. METHODS: In a retrospective study, results of automated static perimetry in the central 10-degree visual field were reviewed for patients with retinitis pigmentosa and choroideremia. Univariate linear regression of mean deviation was undertaken for each eye that had field data from at least five tests in addition to the prior static perimetric experience during a period of 3.5 years or more. Data from 30 eyes of 16 patients fulfilled eligibility requirements; the mean number of fields was 5.8 (range, five to 10) and the mean follow-up period was 64 months (range, 42 to 97 months). Eyes were classified as progressive if the regression coefficient was negative and significantly different from zero, with a P value of less than .05. RESULTS: Among 14 patients in whom both eyes were eligible for univariate linear regression analysis, a statistically significant progression was demonstrated in both eyes in four patients, only in one eye in five patients, and in neither eye in five patients. One of two patients, in whom only one eye was eligible for the study, showed significant progression in the eye. Overall, 14 (47%) of 30 eyes showed statistically significant progression with respect to the mean deviation. No eye showed significant improvement. CONCLUSIONS: Automated static perimetry of the central 10-degree visual field measured approximately once or twice a year demonstrated the progressive nature of the disease with the use of univariate linear regression of mean deviation in 47% of eyes with retinitis pigmentosa and choroideremia during the follow-up period of 3.5 years or more. These results may be useful in understanding the clinical course of the diseases and counseling patients.

Adolescent↗
PubMed1999

Optical coherence tomography of idiopathic polypoidal choroidal vasculopathy.

PURPOSE: To document and study the cross-sectional structures of polypoidal elements using optical coherence tomography in eyes with idiopathic polypoidal choroidal vasculopathy. METHODS: Optical coherence tomography images of two eyes with idiopathic polypoidal choroidal vasculopathy were correlated with slit-lamp biomicroscopic findings, fundus photographs, fluorescein angiograms, and indocyanine green angiograms. RESULTS: Cross-sectional optical coherence tomographic images of retinochoroidal structures showed prominent anterior protrusion of the orange subretinal mass corresponding to the polypoidal structure in the indocyanine green angiogram. Hemorrhagic detachment of the retinal pigment epithelium was contiguous with the cone-shaped nodule beneath the retinal pigment epithelium in one eye, and an apparent discontinuity was observed in the highly reflective layer that delineates the polypoidal structure. CONCLUSIONS: Some of the polypoidal structures in eyes with idiopathic polypoidal choroidal vasculopathy are anteriorly protruding lesions in the inner choroid that may cause serosanguineous detachment of the retinal pigment epithelium through damage of the overlying Bruch membrane, retinal pigment epithelium, and the adhesion between them. Cross-sectional optical coherence tomographic images may increase understanding of the pathophysiology of idiopathic polypoidal choroidal vasculopathy.

Aged↗
PubMed1999

Optical coherence tomography of successfully repaired idiopathic macular holes.

PURPOSE: To present the cross-sectional retinal imaging results of optical coherence tomography in eyes with successfully repaired idiopathic macular hole and their relevance to visual recovery. METHODS: We studied 33 eyes with successful repair of an idiopathic macular hole through vitrectomy and fluid-gas exchange from 32 patients (11 men and 21 women) with ages ranging from 48 to 78 years, with a median age of 66 years. Preoperative conditions in eyes with primary surgery disclosed nine eyes with stage 2, 14 eyes with stage 3, and four eyes with stage 4 macular hole. An additional six eyes underwent a second surgery because the previous surgery was unsuccessful. Measurement of best-corrected visual acuity, slit-lamp biomicroscopy with fundus contact lens, fundus photographs, and optical coherence tomographic examination were performed between 6 and 9 months after surgery in 29 eyes and between 15 and 36 months after surgery in four eyes. RESULTS: Optical coherence tomographic images of the repaired macular holes were categorized into three patterns. U-type (normal foveal contour; 13 eyes) showed mildly to moderately backscattering layers with a smooth circular surface covering retinal pigment epithelium and choriocapillaris layers. In eyes with V-type (steep foveal contour; 13 eyes), the retinal pigment epithelium and choriocapillaris layers were covered with moderately backscattering layers with a notch. W-type (foveal defect of neurosensory retina; seven eyes) showed abruptly or gradually terminating sensory retinal layers to expose the surface of the retinal pigment epithelium and choriocapillaris layers. Postoperative acuity was well correlated with these patterns of optical coherence tomographic images. CONCLUSION: Assessment of successfully repaired idiopathic macular holes with optical coherence tomographic images provides a useful correlation with postoperative visual recovery.

Aged↗
PubMed1999

Hydroxyapatite applied as direct pulp capping medicine substitutes for osteodentin.

Tissue changes in rat incisors after direct pulp capping with hydroxyapatite (OH-AP) have been investigated using light and electron microscopy. Immediately after direct pulp capping, necrotic cells and cell debris were scattered only in the superficial zone of an exposed site. At day 1, both neutrophils having many vacuoles and macrophages phagocytizing numerous OH-AP granules appeared in the lower part of the OH-AP layer. At day 3, the cells surrounding the basal part of the OH-AP layer had large nuclei and well-developed rough-surfaced endoplasmic reticulum. At day 5, the newly formed mineralized tissue consisting of needle-like crystals was observed directly adjacent to the deepest zone of the OH-AP layer. However, fibroblast-like cells phagocytosing applied OH-AP granules were seen even at this period. These findings suggest that the applied OH-AP could directly substitute for osteodentin.

Animals↗
PubMed1999

Targeted disruption of cd39/ATP diphosphohydrolase results in disordered hemostasis and thromboregulation.

CD39, or vascular adenosine triphosphate diphosphohydrolase, has been considered an important inhibitor of platelet activation. Unexpectedly, cd39-deficient mice had prolonged bleeding times with minimally perturbed coagulation parameters. Platelet interactions with injured mesenteric vasculature were considerably reduced in vivo and purified mutant platelets failed to aggregate to standard agonists in vitro. This platelet hypofunction was reversible and associated with purinergic type P2Y1 receptor desensitization. In keeping with deficient vascular protective mechanisms, fibrin deposition was found at multiple organ sites in cd39-deficient mice and in transplanted cardiac grafts. Our data indicate a dual role for adenosine triphosphate diphosphohydrolase in modulating hemostasis and thrombotic reactions.

Adenosine Triphosphatases↗