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Biomedical subjects

H Doi

Publications and source records attributed to H Doi.

At least 55 records · Page 3Linked to original sources
PubMed2000

New technique for producing hybridoma by using laser radiation.

BACKGROUND AND OBJECTIVE: With conventional methods, cell fusion rate is extremely low, and fusion of two specific cells is not possible. We developed a new method for inducing cell fusion under the microscope by using a microprocessing device by laser. STUDY DESIGN/MATERIALS AND METHODS: Under a microscope, the target cells were irradiated with laser beams for trapping. Then, the trapped cells were transferred and placed in contact with the corresponding cells, which were also fixed by laser beam. The pulse laser beams are focused on the contact surface to cut small perforations for mutual communication of the cytoplasms. RESULTS: The fusion rate of mouse myeloma cells was 38%. The rate of hybridoma production of myeloma cell and lymphocyte was 2%. We confirmed the proliferation of the newly formed hybridoma in HAT medium and the production of immunoglobulin G. CONCLUSION: This new cell fusion method is characterized by production of hybridomas of target cells, lower cell toxicity, and a high rate of hybrid production.

Animals↗
PubMed2000

Mechanisms of tubulin modification by phosphatidylcholine hydroperoxides.

The interaction of lipid peroxides with cellular proteins has been postulated to contribute to cellular aging. A potential target for such effects is tubulin, the building block of microtubules. We examined the concentration-dependent effects of phosphatidylcholine hydroperoxides on the ability of tubulin to polymerize into microtubules. The results demonstrated that even very low concentrations of peroxides were sufficient to interfere with the tubulin and, therefore, the microtubule function. Decreased tubulin activity (as measured by tubulin GTPase activity) showed correlation with the modification of methionine and cysteine in tubulin and a change in the tubulin conformational state as indicated by fluorescence and ultraviolet spectroscopic measurements. As no effect on electric conductivity was observed, indicating that modulation of ionic binding was not involved, the interaction mechanism may be a hydrophobic one.

Amino Acids↗
PubMed2000

Comparison of the effect of imidaprilat and ramiprilat on broncho-constriction and hypotension induced by bradykinin in guinea pigs.

OBJECTIVE: To demonstrate tissue selective brady-kinin (BK) potentiating action of angiotensin converting enzyme inhibitors, we studied effects of imidaprilat and ramiprilat, active metabolites of imidapril and ramipril, respectively, on bronchoconstriction and hypotension both induced by BK in vasopressin-infused anesthetized guinea pigs. METHODS: We measured pulmonary inflation pressure and blood pressure in vasopressin-infused anesthetized guinea pigs at the same time. BK-induced changes in pulmonary inflation pressure and blood pressure before and after the administration of ACE inhibitor were compared. RESULTS: Imidaprilat and ramiprilat enhanced BK-induced hypotension comparably, and this effect was inhibited by Nomega-nitro-L-arginin-methylester (L-NAME, a nitric oxide synthetase inhibitor). Although imidaprilat did not affect BK-induced bronchoconstriction, ramiprilat enhanced the broncho-constriction significantly. SR48968, a selective NK2 receptor antagonist, significantly inhibited the enhancing effect of ramiprilat on BK-induced bronchoconstriction. CONCLUSION: These results suggest that enhancement of BK-induced hypotension by imidaprilat and ramiprilat is mediated by nitric oxide (NO), but the mediator of the enhancing action of ramiprilat on BK-induced bronchoconstriction is mainly neurokinin A.

Angiotensin I↗
PubMed2000

Structural and functional study of K453E mutant protective protein/cathepsin A causing the late infantile form of galactosialidosis.

To clarify the molecular basis of the late infantile form of galactosialidosis, we characterized a defective protective protein/cathepsin A (PPCA) gene product with the K453E mutation newly found in an Arabic patient with this disease. Immunocytochemical, expression, and metabolic studies revealed that the precursor PPCA was synthesized but not processed to the mature form, and it was degraded in the mutant. A structural model of the mutant PPCA was constructed by amino acid substitution of 453glutamic acid for lysine in the crystal structure of the wild type PPCA precursor reported. The results show that the K453E mutation is located at the dimer interface of the PPCA and reduces the hydrogen bond formation in the dimer. This structural change may cause instability of the PPCA dimer.

Amino Acid Substitution↗
PubMed2000

Molecular and structural studies of Japanese patients with sialidosis type 1.

To gain insight into the pathogenesis of sialidosis type 1, we performed molecular investigations of two unrelated Japanese patients. Both of them are compound heterozygotes for base substitutions of 649G-to-A and 727G-to-A, which result in amino acid alterations V217M and G243R, respectively. Using homology modeling, the structure of human lysosomal neuraminidase was constructed and the structural changes caused by these missense mutations were deduced. The predicted change due to V217M was smaller than that caused by G243R, the latter resulting in a drastic, widespread alteration. The overexpressed gene products containing these mutations had the same molecular weight as that of the wild type, although the amounts of the products were moderately decreased. A biochemical study demonstrated that the expressed neuraminidase containing a V217M mutation was partly transported to lysosomes and showed residual enzyme activity, although a G243R mutant was retained in the endoplasmic reticulum/Golgi area and had completely lost the enzyme activity. Considering the data, we surmise that the V217M substitution may be closely associated with the phenotype of sialidosis type 1 with a late onset and moderate clinical course.

Adult↗
PubMed2000

Increase in plasma levels of secretory type II phospholipase A(2) in patients with coronary spastic angina.

OBJECTIVE: Plasma levels of sPLA(2) were increased in various chronic inflammatory diseases including coronary artery disease. Lipid products mediated through PLA(2) have been shown to induce impairment of endothelium-dependent dilation, contraction of smooth muscle and proliferation of smooth muscle cells, all of which might lead to coronary spasm. Thus, this study investigated whether plasma levels of secretory non-pancreatic type II phospholipase A(2) (sPLA(2)) may be increased in patients with coronary spastic angina, considering the possible link of sPLA(2) with pathogenesis of coronary artery spasm. METHODS: Plasma levels of sPLA(2) in peripheral circulation, in coronary sinus and in aortic root were measured in 57 patients with coronary spastic angina, 46 patients with stable effort angina and 53 control patients by radioimmunoassay. RESULTS: The peripheral plasma levels of sPLA(2) were increased in patients with coronary spastic angina compared with control patients. In multivariate statistical analysis, the increase in sPLA(2) levels was a significant risk for the presence of coronary spasm independent of other risk factors including C-reactive protein levels. The coronary sinus-arterial difference of plasma sPLA(2) levels, reflecting sPLA(2) released into the coronary circulation, was increased during coronary spasm induced by the intracoronary infusion of acetylcholine in patients with coronary spastic angina, but it remained unchanged both during the acetylcholine infusion and during myocardial ischemia provoked by rapid atrial pacing in patients with stable effort angina and in control patients. CONCLUSION: The increase in peripheral plasma levels of sPLA(2) is a significant risk factor for the presence of coronary spasm and it may possibly reflect inflammatory activity in spasm coronary arteries.

Acetylcholine↗
PubMed2000

Tensile properties and transformation temperatures of Pd added Ti-Ni alloy dental castings.

The effect of palladium (Pd) addition to Ti-Ni alloy as the third element was investigated to improve the super-elasticity of the alloy castings at body temperature for dental application. Ti-50.8Ni (at %) alloy, which exhibited super-elasticity at 310 K in castings, was used for comparison. 5.0, 7.5, 10.0 and 15.0 at % Pd was added to Ti-50.0Ni alloy by the substitution for Ni. The change in the proportion of Ti and Ni was also examined at the fixed Pd addition of 7.5 at %. The properties of the alloys were investigated in tensile test and differential scanning calorimetry (DSC). Ti-42.5Ni-7.5Pd alloy castings showed good super-elasticity among the examined alloys from the viewpoint of residual strain and elongation. Moreover, apparent proof stress could be changeable by the proportion of Ti and Ni with residual strain being kept low. Ti-42.5Ni-7.5Pd alloy castings exhibited better super-elastic flexibility than Ti-50.8Ni alloy, which is proven by lower apparent proof stress and larger elongation. This flexibility appears to be caused by its relatively high martensitic transformation starting temperature point. It is suggested that this flexibility with super-elasticity could widen the clinical application of the alloy casting in dentistry.

Journal Article↗
PubMed2000

Stress transmission through Ti-Ni alloy, titanium and stainless steel in impact compression test.

Impact stress transmission of Ti-Ni alloy was evaluated for biomedical stress shielding. Transformation temperatures of the alloy were investigated by means of DSC. An impact compression test was carried out with use of split-Hopkinson pressure-bar technique with cylindrical specimens of Ti-Ni alloy, titanium and stainless steel. As a result, the transmitted pulse through Ti-Ni alloy was considerably depressed as compared with those through titanium and stainless steel. The initial stress reduction was large through Ti-Ni alloy and titanium, but the stress reduction through Ti-Ni alloy was more continuous than titanium. The maximum value in the stress difference between incident and transmitted pulses through Ti-Ni alloy or titanium was higher than that through stainless steel, while the stress reduction in the maximum stress through Ti-Ni alloy was statistically larger than that through titanium or stainless steel. Ti-Ni alloy transmitted less impact stress than titanium or stainless steel, which suggested that the loading stress to adjacent tissues could be decreased with use of Ti-Ni alloy as a component material in an implant system.

Journal Article↗
PubMed2000

Hyperbaric oxygen (HBO) therapy for acute traumatic cervical spinal cord injury.

STUDY DESIGN: A retrospective study of spinal cord injury (SCI) treated with and without hyperbaric oxygen (HBO) therapy. OBJECTIVES: To report on the use of HBO in spinal cord injury. SETTING: Neurosurgical Unit, Tokyo, Japan. METHODS: Thirty-four cases of hyperextension spinal cord injury without bone damage and previous history of surgical intervention were divided into two groups, with (HBO) or without (non-HBO) therapy. The neurological findings at admission and their outcomes were evaluated by means of Neurological Cervical Spine Scale (NCSS)1 and the average improvement rates in individual groups were compared. RESULTS: The improvement rate ranged from 100% to 27.3% with the mean value of 75. 2% in the HBO group, while these values were 100%, 25.0% and 65.1% respectively in the non HBO group. CONCLUSION: In the HBO group, the improvement rate indicated effectiveness in acute traumatic cervical spinal cord injury.

Adult↗
PubMed2000

Lysophosphatidylcholine induces urokinase-type plasminogen activator and its receptor in human macrophages partly through redox-sensitive pathway.

Urokinase-type plasminogen activator (uPA) and its cell surface receptor (uPAR) have been shown to be expressed in macrophages in atherosclerotic arterial walls, but the regulatory mechanisms of their expression remain unclear. The present study was performed to examine the effects of lysophosphatidylcholine (lysoPC), an important atherogenic lipid, on the expression of uPA and uPAR in human monocyte-derived macrophages. LysoPC upregulated the mRNA expression of uPA and uPAR, and it increased the protein expression of uPA in the culture medium and bound to the cell surface and of uPAR in the particulate fraction of the cells. LysoPC significantly increased the binding of the amino-terminal fragment of uPA to the treated cells and the cell-associated plasminogen activator activity. LysoPC stimulated superoxide anion production and increased intracellular oxidant levels in the cells. The combined incubation with reduced glutathione diethyl ester or N-acetylcysteine, antioxidants, suppressed the upregulation of uPA and uPAR mRNA and the increase in plasminogen activator activity by lysoPC. uPA and uPAR mRNA expression was also induced by the incubation with xanthine and xanthine oxidase, a superoxide anion-generating system. The results suggest that lysoPC increased the expression of uPA and uPAR and their functional activities in human monocyte-derived macrophages, at least in part through a redox-sensitive mechanism. This coordinate increase in the expression of uPA and uPAR in human macrophages by lysoPC could play an important role in plaque formation and disruption, arterial remodeling, and angiogenesis in atherosclerotic arterial walls.

Acetylcysteine↗
PubMed2000

Super-elastic property of Ti-Ni alloy for use in dentistry.

The super-elasticity of Ti-Ni alloy was investigated in tensile and bending tests to evaluate the mechanical properties of the castings and heat treatment effect on the bending properties of the wires for new clinical applications of the alloy in prosthodontics and orthodontics. In terms of the tensile properties of the castings, apparent proof stress increased and elongation decreased with the small increase in nickel content or with the decrease in titanium purity. This result suggests that precise control of the materials is indispensable to utilize the super-elasticity in dental cast appliances. In terms of the bending properties of Ti-Ni alloy wires, low residual deflection and small load/deflection ratio were obtained by the second heat treatment between 733 and 813 K, which was found to be suitable for the shape memory treatment of orthodontic appliances. Moreover, the functional force was changeable within this treatment temperature range.

Biocompatible Materials↗
PubMed2000

Large-scale cDNA analysis reveals phased gene expression patterns during preimplantation mouse development.

Little is known about gene action in the preimplantation events that initiate mammalian development. Based on cDNA collections made from each stage from egg to blastocyst, 25438 3'-ESTs were derived, and represent 9718 genes, half of them novel. Thus, a considerable fraction of mammalian genes is dedicated to embryonic expression. This study reveals profound changes in gene expression that include the transient induction of transcripts at each stage. These results raise the possibility that development is driven by the action of a series of stage-specific expressed genes. The new genes, 798 of them placed on the mouse genetic map, provide entry points for analyses of human and mouse developmental disorders.

Animals↗
PubMed2000

Unclassified connective tissue disease presenting as cardiac tamponade: a case report.

This report describes a case of cardiac tamponade as the initial manifestation of unclassified connective tissue disease (UCTD). A 68-year-old Japanese woman was admitted to hospital because of dyspnea and edema. She had undergone a radical left mastectomy for the treatment of breast cancer 18 years before. On admission, bilateral leg edema, hepatomegaly, and a paradoxical pulse were noted on physical examination. The erythrocyte sedimentation rate was elevated and the C-reactive protein was 2.8 mg/dl. Antinuclear antibodies and anti-SS-A/Ro antibodies were present. The scl-70 and anticentromere antibodies were elevated. Chest radiography showed cardiomegaly. Echocardiography revealed a large pericardial effusion, but the pericardial fluid did not contain malignant cells or bacteria. She did not meet the diagnostic criteria for any known connective tissue diseases, so was diagnosed with cardiac tamponade due to UCTD. Prednisolone (30 mg/day) was administered, which resulted in a gradual resolution of the pericardial effusion. Although connective tissue diseases are known to cause pericardial effusion, cardiac tamponade as the initial manifestation of the disease in the absence of other symptoms is quite rare.

Aged↗
PubMed2000

Protective effect of TA-993, a novel therapeutic agent for peripheral circulatory insufficiency, on skeletal muscle fatigue in a rat model of hindlimb ischemia.

TA-993 (cis-(-)-2-(4-methylphenyl)-3-acetoxy-2,3-dihydro-5-(2-dimethylaminoethy l)-8-methyl1,5-benzothiazepine-4(5H)-one maleate), a new 1,5-benzothiazepine derivative, has a selective increasing action on limb blood flow in addition to an antiplatelet action. In this report we studied the effect of TA-993 on a time dependent decrease in developed tension of electrically-induced contraction of tibialis anterior muscle in a rat model of peripheral circulatory insufficiency induced by occlusion of abdominal aorta. In our preparation, the developed tension decreased by 20-30% in a sham-operated group and 30-40% in an abdominal aorta-occluded group at the end of the experimental period of 60 min. Intraduodenal administration (i.d.) of TA-993 (10 mg/kg) to the abdominal aorta-occluded rats ameliorated the decrease in developed tension to the level of the sham-operated group. Moreover, TA-993 at 10 mg/kg, i.d. significantly increased femoral arterial blood flow supplied through collateral circulation and decreased the whole blood viscosity in this model. These results suggest that TA-993 improves dysfunction of skeletal muscle contraction due to peripheral circulating insufficiency through an increase in collateral blood flow and an improvement of red blood cell deformability.

Animals↗