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Biomedical subjects

K Takeuchi

Publications and source records attributed to K Takeuchi.

At least 883 records · Page 49Linked to original sources

The poliovirus receptor protein is produced both as membrane-bound and secreted forms.

Both genomic and complementary DNA clones encoding poliovirus receptors were isolated from genomic and complementary DNA libraries prepared from HeLa S3 cells, respectively. Nucleotide sequence analysis of these cloned DNAs revealed that the poliovirus receptor gene is approximately 20 kb long and contains seven introns in the coding region, and that at least four mRNA isoforms referring to the coding sequence are generated by alternative splicing and appear to encode four different molecules, that is, PVR alpha, PVR beta, PVR gamma and PVR delta. The predicted amino acid sequences indicate that PVR alpha and PVR delta, corresponding to the previously described cDNA clones H20A and H20B, respectively, are integral membrane proteins while the other two molecules described here for the first time lack a putative transmembrane domain. Mouse cell transformants carrying PVR alpha were permissive for poliovirus infection, but those carrying PVR beta were hardly permissive. In contrast to PVR alpha, PVR beta was not detected on the surface of the mouse cell transformants but was detected in the culture fluid by an immunological method using a monoclonal antibody against poliovirus receptor. Three types of splicing products for PVR alpha, PVR beta and PVR gamma were detected by polymerase chain reactions using appropriate primers in poly(A)+ RNAs of the brain, leukocyte, liver, lung and placenta of humans; the choice of primers used did not permit detection of PVR delta. In situ hybridization using a cDNA fragment as a probe demonstrated that the PVR gene is located at the band q13.1----13.2 of human chromosome 19.

Amino Acid Sequence↗

The effect of middle ear effusions from children on in vitro ciliary activity.

In an attempt to clarify the effect of middle ear effusions (MEEs) on ciliary activity, the ciliary beat frequency (CBF) of nasal epithelium from normal subjects was determined following incubation in Hanks' solution containing effusions from pediatric otitis media with effusion. Both serous and mucoid types of effusions increased CBF 15 min after incubation. The mucoid type of effusion was then found to decrease CBF 6 h after incubation. Since the pH and osmotic pressure of the solution containing the effusions were within the physiological ranges for normal ciliary beating, our findings show that MEEs can influence CBF in vitro.

Child↗

Viscoelastic properties of middle ear effusions from pediatric otitis media with effusion and their relation to gross appearance.

The viscosity and the elasticity were measured in 146 individual middle ear effusions (MEEs) from 93 children with otitis media with effusion (OME). The effusions were classified into serous or mucoid type by their macroscopic appearance. The distribution of elasticity showed two peaks, corresponding to the peaks of serous and mucoid types of effusions. There were a small number of effusions in the intermediate range of elasticity, where the effusions were most effectively transported by the cilia. These findings suggests that the MEEs of pediatric OME are accumulations of rheologically suboptimal fluid in the middle ear cavity.

Child↗

Nucleotide sequence of the gene encoding the matrix protein of mumps virus (Miyahara strain).

The nucleotide sequence of the gene encoding the matrix (M) protein of mumps virus (MuV), Miyahara strain, has been determined from several overlapping cDNA clones. The M protein mRNA is 1248 nucleotides in length, exclusive of the poly(A) tail, and codes for a protein of 375 amino acids (Mr41,556). Comparison of the deduced amino acid sequence of the M protein of the Miyahara strain with that of the SBL-1 strain revealed that the M proteins of both strains are highly conserved. A significantly lower rate of nucleotide differences conducive to amino acid differences in the M gene compared with other genes appeared to indicate the importance of the conserved primary structure of the M protein for its function.

Amino Acid Sequence↗

Different effects of cytoprotective drugs on ethanol- and aspirin-induced gastric mucosal injury in pylorus-ligated rats.

In anesthetized rats oral administration (2 ml) of both ethanol (50% in 150 mM HCl) and aspirin (80 mM in 150 mM HCl) produced bandlike lesions in the stomach, while more generalized lesions occurred in the pylorus-ligated stomach when the irritant was given intragastrically through the fistula prepared in the rumen and the mucosal folds were removed by stomach distension. The bandlike lesions induced in the intact stomach by both irritants were significantly and dose-dependently prevented by 16,16-dimethyl PGE2 (dmPGE2: 3 and 10 micrograms/kg, subcutaneously), cysteamine (30 and 100 mg/kg, subcutaneously) or timoprazole (10 and 30 mg/kg, per os) at the doses which significantly inhibited gastric motility. In the pylorus-ligated stomach, however, neither of these agents showed any protection against the generalized lesions induced by ethanol, but such lesions caused by aspirin were significantly prevented only by dmPGE2. These agents also showed similar effects against the reduction of transmucosal PD in the pylorus-ligated stomach exposed to ethanol and aspirin. These results suggest that (1) the formation of bandlike lesions caused by ethanol and aspirin depends on the presence of mucosal folds and may be prevented by the agents that inhibit gastric motility, (2) the pathogenesis of the lesions induced by aspirin and ethanol may be different in the pylorus-ligated stomach, and (3) dmPGE2 has a unique protective ability that is not shared by usual cytoprotective agents.

16,16-Dimethylprostaglandin E2↗

Body temperature-dependent action of baclofen in rat stomach. Relation to acid secretion and ulcerogenicity.

The effect of baclofen (PCPGABA) on acid secretion, motility, and mucosa was investigated in the anesthetized rat stomach under various body temperatures (BT: 28-38 degrees C), and they were compared with those of 2-deoxy-D-glucose (2DG), an acid stimulant through cytoglycopenia. Under these conditions PCPGABA induces lesions dose-dependently (greater than 1 mg/kg, subcutaneously) in both the stomach and duodenum, and this action was dependent on BT; lowering of BT enhanced the ulcerogenicity. PCPGABA (3 mg/kg) had no effect on acid secretion at higher BT (36-38 degrees C) but produced a marked increase of acid output at lower BT (30-32 degrees C). 2DG caused a stimulation of acid output and gastric lesions without BT dependency, but the duodenal ulcerogenicity enhanced at lower BT. Gastric motility was enhanced significantly by these two agents to similar degrees, at either high or low BT. Neither PCPGABA nor 2DG affected alkaline secretion in the duodenum, while lowering of BT by itself reduced alkaline secretory responses. The above changes caused by PCPGABA and 2DG were blocked by both atropine and vagotomy. These results suggest that (1) acid stimulatory and ulcerogenic action of PCPGABA may involve a temperature-dependent process but does not relate to a cytoglycopenia, and (2) the vagus nerve mediating acid secretion and motility may be different in the temperature dependency.

Animals↗

Possible mechanisms involved in gastric hypermotility caused by indomethacin in the rat. Role of glycoprivic response.

Pathogenesis of indomethacin-induced gastric lesions was investigated in the rat by measuring lesions, gastric motility, and terminal blood glucose levels and correlating them with each other. Subcutaneously administered indomethacin (3-25 mg/kg) dose-dependently produced lesions in the stomach with concomitant gastric hypermotility and reduction of blood glucose levels. When the lesion score and the motility were plotted against terminal glucose levels, a highly significant relationship was found among these three factors (P less than 0.01). Gastric lesions and hypermotility induced by indomethacin (25 mg/kg) were suppressed significantly by 16,16-dmPGE2 (10 micrograms/kg) with no effect on the glucose levels, while intravenous infusion of glucose (25% w/w, 1.4 ml/hr) prevented these responses and restored the reduced glucose levels above the basal values. In addition, both 16,16-dmPGE2 and glucose infusion afforded a significant protection against gastric lesions induced by indomethacin even in the acid-perfused stomach (150 mM HCl). These results confirmed gastric hypermotility as a key element in the pathogenesis of indomethacin-induced lesions and further suggested that indomethacin may sensitive gastric contractility through glycoprivic receptors by inducing hypoglycemia and PG deficiency.

16,16-Dimethylprostaglandin E2↗

Effect of dimethadione derived from repeated oral administration of trimethadione on pancreatic secretion in dogs.

The effect of the weak organic acid of dimethadione (DMO) on secretin-stimulated pancreatic secretion was studied with repeated oral administration of trimethadione (TMO), the precursor of DMO, to dogs at a dose of 10 to 160mg/kg/day for a period of 14 days. The bicarbonate concentration in pancreatic juice at a steady state decreased significantly, reflecting a close correlation with the dose of TMO and DMO concentrations in plasma and pancreatic juice. The maximal decrement from the control of cases of no TMO administration was 18.8 mEq/l (12.1% of the control level). The chloride concentration in pancreatic juice showed a reciprocal relation to the bicarbonate concentration. The sum of both anion concentration was constant, irrespective of the dose of TMO. The average carbon dioxide tension of pancreatic juice in all doses of TMO was lower than that of the control, but differences were not statistically significant. The pH, flow rate, sodium and potassium concentrations in pancreatic juice at a steady state did not differ significantly in relation to the dose of TMO. These findings suggest that repeated oral administration of TMO cause a significant decrease in bicarbonate concentration in pancreatic juice, resulting probably from the buffer action of bicarbonate on protons provided from the undissociated form of DMO.

Administration, Oral↗

Actin cytoskeleton of resting bovine platelets.

Actin filaments in resting discoid bovine platelets were examined by fluorescence and electron microscopy. Rhodamine-phalloidin staining patterns showed a characteristic wheel-like structure which consisted of a central small circle connected by several radial spokes to a large peripheral circle. This wheel-like structure was composed of actin filaments forming a characteristic arrowhead structure with heavy meromyosin from muscle. Actin filaments were densely arrayed in parallel with a marginal microtubule band and radiated out from the center to the periphery. Platelets treated with colchicine lost their marginal microtubule band but retained their wheel-like structure and normal discoid form. Cytochalasin B disrupted the wheel-like structure but not the marginal microtubule band or the normal discoid form. After simultaneous treatment with both cytochalasin B and colchicine, platelets lost their discoid shape. These results suggest that actin filaments and microtubules both play important roles in the maintenance of the discoid shape of resting bovine platelets.

Actins↗

Detection and characterization of mumps virus V protein.

By cDNA cloning, DNA sequencing, and RNAse mapping analyses two distinct mRNA species were shown to be transcribed from the mumps virus P gene; one faithful and the other edited copies. The former was found to direct the synthesis of the V protein (25K), while the latter, after the addition of two nontemplated G residues, was found to direct the synthesis of the P protein (40K-42K). The carboxy terminal of the V protein contained a cysteine-rich region which was similar but not identical to the metal binding domain motif found in several nucleic acid binding proteins. The V protein was detected not only in mumps virus-infected cells but also in the virions by antiserum raised against a synthetic peptide.

Amino Acid Sequence↗

Nucleotide sequence of the leader and nucleocapsid protein gene of mumps virus and epitope mapping with the in vitro expressed nucleocapsid protein.

The nucleotide sequence of the leader and the gene encoding the nucleocapsid protein (NP) of mumps virus Miyahara strain have been determined. The leader sequence is 55 nucleotides in length and the NP gene is 1845 nucleotides in length, exclusive of poly(A). The NP gene codes for a protein of 549 amino acids, with a calculated molecular weight of 61,365. For epitope mapping, a series of NPs from which C-termini were serially deleted were expressed in vitro from five mRNA constructs and were examined by radioimmunoprecipitation assay (RIPA) with eight nonoverlapping monoclonal antibodies (MoAbs) against the mumps virus NP. It was found that seven out of eight MoAbs reacted with the NP synthesized in vitro. Five recognized the epitopes located within the C-terminal 74 amino acids region and one within the adjacent 64 amino acids upstream. The epitope of the remaining one was in the N-terminal half of the NP.

Amino Acid Sequence↗

Determination of gastroduodenal alkaline responses in the rat.

We set up a new system for measuring the gastroduodenal HCO3- responses using pH change and potential difference (PD) in the anesthetized rat. The stomach or the proximal duodenum was perfused at the flow rate of 0.7 mL/min with saline (pH 4.5), the pH of the perfusate and PD were continuously monitored, and HCO3- output was determined by back-titrating the perfusate and by measuring the area of pH change. In the case of the stomach, acid secretion was inhibited by omeprazole (60 mg/kg, i.p.). Output of both pH and HCO3- in these tissues was significantly increased by intravenous administration of prostaglandin (PGE2), 16, 16-dimethyl PGE2, carbachol, and YM-14673 (a thyrotropin-releasing hormone (TRH) analog), whereas the PD responded to these agents by a significant rise in the duodenum and decrease in the stomach. These parameters also responded to physiological stimulation such as mucosal acidification. When the area of pH change caused by various agents was plotted against the net amount of HCO3- output determined from back-titration, a significant relationship was found between these two factors (r = 0.98). These results indicate that this system using pH change may be useful for quantitative determination of HCO3- response in the gastroduodenal mucosa.

Animals↗

Differentiation of the mumps vaccine strains from the wild viruses by the nucleotide sequences of the P gene.

Nucleotide sequence analysis of a part of the P gene of mumps virus allowed the differentiation of live attenuated mumps vaccine strains from each other and from wild mumps viruses. Restriction enzyme analysis was found to serve as a convenient method of screening for one strain of mumps vaccine. Examination by the method of virus isolates obtained from the patients who developed mumps parotitis or meningitis following vaccination revealed that most of those viruses were related to the respective vaccine viruses used for immunization.

Base Sequence↗

Circulating immunoreactive endothelin in ischemic heart disease.

Circulating immunoreactive endothelin (ir-ET) was measured in nine patients with acute myocardial infarction (AMI), 10 patients with stable angina pectoris (SAP), and 25 normal control subjects. In patients with AMI, the plasma ir-ET level was elevated in the acute phase and was highest on the day of onset (AMI: 3.8 +/- 1.7 pg/ml, normal control value: 0.5 +/- 0.2 pg/ml). The plasma ir-ET level showed a positive correlation with the wall motion abnormality index (rs = 0.56, p less than 0.01), thrombin-antithrombin III complex (rs = 0.55, p less than 0.01), and beta thromboglobulin (rs = 0.39, p less than 0.05). An especially high plasma ir-ET level was detected in patients in whom the Killip subset was IV. The plasma ir-ET level was not increased in patients with SAP (0.8 +/- 0.3 pg/ml). The plasma ir-ET level is increased in the acute phase of AMI. A pathophysiologic state characterized by cardiac dysfunction, an activated coagulation system, and platelet hyperactivity may be associated with this increase in plasma ir-ET.

Angina Pectoris↗

Effect of the secretions from the IUD-bearing uterus on peri-implantation mouse embryos.

This study was performed to evaluate the effect of IUD-bearing uterine secretions on peri-implantation mouse embryo. 83% of blastocysts degenerated by 96 hr after co-culture with uterine fluids from the IUD-bearing uterus. This degeneration rate was contrasted to 78% after heat treatment (56 degrees C for 30 min) of the IUD-bearing uterine secretions. These results suggest that embryotoxic agents are present in the fluid of the IUD-bearing uterus, especially in the supernatant component and that these agents may be related to the presence of macrophage or a chemical mediator produced by macrophage.

Animals↗

A therapy against vasospasm after subarachnoidal haemorrhage: clinical experience of balloon angioplasty.

Vasospasm is a phenomenon often present in human cases after subarachnoidal haemorrhage. Its aetiology and pathophysiology are unknown. As a result, cases of vasospasm do not respond to any known pharmacological therapy. We present in this paper, a practical application of intravascular surgery for vasospasm. In this procedure the requirement for a portable digital subtraction unit, transcranial Doppler sonography, and being able to determine quickly the neurological state of the patient are very important points. The use of monitoring equipment is especially indispensable during the carrying out of percutaneous transluminal angioplasty (PTA), which should be performed as soon as possible after the occurrence of vasospasm. The case of a 54-year-old male patient suffering from a ruptured anterior cerebral artery aneurysm was studied. Neck clipping was performed on the second day. On the fifth day the neurological state was stupor, and transcranial Doppler sonography (TCD) revealed a flow velocity of over 100 cm s-1. PTA was performed on the sixth day. The bilateral middle cerebral artery was dilated and the flow velocity slowed to within the normal range. The neurological state improved to clouding of consciousness after PTA. On the eighteenth day, the patient's consciousness and bilateral carotid angiograms revealed the vessels to be dilated still. It was concluded that PTA therapy, and the use of a portable digital subtraction unit and TCD sonography, for vasospasm caused by subarachnoidal haemorrhage is useful and effective.

Angioplasty, Balloon↗

Transcranial Doppler monitoring in severe brain damage: relationships between intracranial haemodynamics, brain dysfunction and outcome.

Transcranial Doppler (TCD) monitoring in the middle cerebral (MCA) and common carotid arteries (CCA) was studied in 105 comatose patients with severe brain damage. TCD-measured velocity waveforms in the MCA were evaluated in relation to loss of cerebral function as assessed by EEG and compressed spectral arrays, loss of brain stem function as measured by somatosensory and auditory evoked potentials, loss of all brain function and the clinical outcome. The velocity waveforms exhibited six patterns: continuous forward flow (FF); diastolic no flow (NF); diastolic reverse flow with (RF) or without (DRF) diastolic FF; brief systolic FF (SFF); and undetectable (U). In the 58 fatal cases, the appearance of RF/DRF or SFF in the MCA more often preceded loss of brain stem function than loss of cerebral function. A U pattern in the MCA, which was confirmed by loss of FF in the CCA, was correlated with loss of brain stem function. Only one of the 47 survivors showed DRF in the MCA. No patients in whom SFF or U was observed in the MCA survived. Therefore the presence of these patterns is reliably predictive of brain death. Intracranial diastolic reverse flow (DRF), however, indicates imminent loss of brain function and the need for prompt resuscitative measures.

Adolescent↗

Chronic effects of synthetic endothelin on blood pressure and sodium excretion in rats.

Continuous infusion of endothelin at a rate of 60 micrograms/kg/day into the jugular vein of rats via osmotic minipumps induced a significant increase in systolic blood pressure, but did not induce any significant changes in urine volume and urinary sodium excretion, compared to those in vehicle-infused rats. When 6 mg/kg/day of benidipine, a newly developed calcium channel blocker, was administered simultaneously with 60 micrograms/kg/day of endothelin, the systolic blood pressure of conscious rats rose on Day 1 to only 137.0 +/- 2.4 mm Hg (P less than .05) compared to the rise to 163.8 +/- 4.7 mm Hg when endothelin alone was infused. The antihypertensive effect of benidipine was sustained. The present results suggest that endothelin can act as a circulating hormone. In addition, they clearly demonstrate that the calcium channel blocker attenuates the elevation of blood pressure induced by endothelin.

Animals↗