PubMed HealthSearch

Biomedical subjects

H Tabuchi

Publications and source records attributed to H Tabuchi.

10 recordsLinked to original sources

Truncation of N-terminal extracellular or C-terminal intracellular domains of human ETA receptor abrogated the binding activity to ET-1.

We have investigated the function of N-terminal and C-terminal domains of the human ETA receptor by expressing truncated mutants in COS-7 cells. Three kinds of ETA receptors truncated in the N-terminal extracellular or C-terminal intracellular domains were produced. Deletion of the entire extracellular N-terminal or intracellular C-terminal domain completely inactivated the ET-1 binding activity. However, the deletion of one half of the N-terminal extracellular domain of the ETA receptor, missing one of two N-linked glycosylation sites, maintained complete binding activity. Specific monoclonal antibodies detected all the truncated ETA receptors in the cell membrane fraction of transfected COS-7 cells. The size of the ETA receptor was heterogeneous due to differential glycosylation and distributed in 48K, 45K and 42K dalton bands in Western blot analysis. These results demonstrated that a part of the N-terminal domain in close proximity to the first transmembrane region is required for the ligand binding activity of the ETA receptor, and the C-terminal domain is perhaps necessary as an anchor for maintenance of the binding site.

Amino Acid Sequence

DNA supercoiling facilitates the assembly of transcriptionally active chromatin on the adenovirus major late promoter.

Assembly of nucleosomes on the adenovirus major late promoter blocked initiation of transcription by RNA polymerase II. However, the formation of transcription preinitiation complexes prevented subsequent assembly of promoter sequences into nucleosomes and allowed transcription on the chromatin templates. When the formation of preinitiation complexes was in competition with nucleosome assembly, transcription on linear or relaxed closed circular DNA was inactivated by nucleosome assembly over the promoter region. However, transcription on partially supercoiled DNA (mean superhelical density of -0.036) remained active because the rapid formation of preinitiation complexes prevented subsequent assembly of promoter sequences into nucleosomes.

Adenoviridae

Robot system for preparing lymphocyte chromosome.

Towards the automatization of the scoring of chromosome aberrations in radiation dosimetry with the emphasis on the improvement of biological preparations, the conventional culture and harvesting method was modified. Based on this modified method, a culture and harvest robotic system (CHROSY) for system (CHROSY) for preparing lymphocyte chromosome was developed. The targeted points of the modification are as in the following. 1. Starting culture with purified lymphocytes in a fixed cell number. 2. Avoiding the loss of cells in changing the liquids following centrifugalization. 3. Keeping the quantity of the liquids to be applied to the treatments of cells fixed. 4. Building a system even a beginner can handle. System features are as follows. 1. Operation system: Handling robot having 5 degrees of freedom; a rotator incubator with an automatic sliding door; units for setting and removing pipette tips; a centrifuge equipped with a position adjuster and an automatic sliding door; two aluminum block baths; two nozzles as pipettes and aspirators connected to air pumps; a capping unit with a nozzle for CO2 gas; a compressor; and an air manipulated syringe. 2. Control system: NEC PC-9801RX21 with CRT; and program written in Basic and Assembly languages on MS-DOS. It took this system 2 hours and 25 minutes to harvest 2 cultures. A fairly good chromosome slide was made from the sample harvested by CHROSY automatically.

Chromosomes

[Left ventricular volume decrease during isovolumic relaxation period in gated blood pool scintigraphy: observations in patients with old myocardial infarction].

We studied left ventricular (LV) volume decrease, namely, the downward displacement of the LV volume curve, during the isovolumic relaxation period on the time-activity curves obtained from gated blood pool scintigraphy in patients with old myocardial infarction (OMI). To evaluate the mechanism and clinical significance of this phenomenon, 113 consecutive patients with OMI undergoing gated blood pool scintigraphy, left ventriculography, and Doppler echocardiography were studied. 1. This phenomenon was observed only in patients with anterior OMI (13 of 51 patients: 25%). Presence (Group I) or absence (Group II) of this phenomenon was examined. 2. On left ventriculography, dyskinetic or aneurysmal wall motion was observed in the anterior or apical region more frequently in Group I (11 of 13 patients: 85%) than in Group II (20 of 51 patients: 39%) (p < 0.001). 3. Doppler echocardiography showed that the presence of abnormal LV reversed flow over 20 cm/sec from the apex to the base during the isovolumic relaxation period is more frequent in Group I (7 of 13: 54%) than in Group II (4 of 51: 8%) (p < 0.001). These results suggested that this blood shift in the left ventricle is attributed to asynchronous LV relaxation occurring simultaneously with LV volume decrease on gated blood pool scintigraphy. In conclusion, this phenomenon suggests the presence of asynchronous LV relaxation.

Echocardiography, Doppler

[A new method for endotracheal intubation with the Bullard laryngoscope].

The Bullard laryngoscope is an anatomically shaped rigid fiber optic instrument designed for indirect laryngoscopy and intubation. It requires no neck extension nor flexion to perform laryngeal intubation. This characteristic is especially useful in the case of difficult airway. The Bullard laryngoscope, however, with which it is easy to visualize the cords, has not been popular yet, because using the intubating forceps mechanism requires a certain amount of skill. The intubation method developed by us with Bullard laryngoscope has been tried on 26 patients with difficult airway and ten patients with normal airway. An endotracheal tube is inserted through nostril, and intubation is performed viewing the tube and larynx during all the processes of intubation. Three kinds of endotracheal tubes, i.e. Magill type plain tubes, styletted tubes, and directional tip tubes, are compared. The method using a directional tip tube (Endotrol) is concluded as the best among them. The Endotrol tube itself has such a suitable shape for nasal intubation that it can be introduced to the larynx with little directional change. Therefore, our method is mastered with a shorter training period than the intubation method with fiber-optic laryngoscopes. In conclusion, our intubation method with the Bullard laryngoscopes using the directional tip tubes (Endotrol) is useful for patients with difficult airways, and is also nontraumatic and easy to perform.

Humans

Recombinant 55-kDa tumor necrosis factor (TNF) receptor. Stoichiometry of binding to TNF alpha and TNF beta and inhibition of TNF activity.

The extracellular domain of the 55-kDa TNF receptor (rsTNFR beta) has been expressed as a secreted protein in baculovirus-infected insect cells and Chinese hamster ovary (CHO)/dhfr- cells. A chimeric fusion protein (rsTNFR beta-h gamma 3) constructed by inserting the extracellular part of the receptor in front of the hinge region of the human IgG C gamma 3 chain has been expressed in mouse myeloma cells. The recombinant receptor proteins were purified from transfected cell culture supernatants by TNF alpha- or protein G affinity chromatography and gel filtration. In a solid phase binding assay rsTNFR beta was found to bind TNF alpha with high affinity comparable with the membrane-bound full-length receptor. The affinity for TNF beta was slightly impaired. However, the bivalent rsTNFR beta-h gamma 3 fusion protein bound both ligands with a significantly higher affinity than monovalent rsTNFR beta reflecting most likely an increased avidity of the bivalent construct. A molecular mass of about 140 kDa for both rsTNFR beta.TNF alpha and rsTNFR beta.TNF beta complexes was determined in analytical ultracentrifugation studies strongly suggesting a stoichiometry of three rsTNFR beta molecules bound to one TNF alpha or TNF beta trimer. Sedimentation velocity and quasielastic light scattering measurements indicated an extended structure for rsTNFR beta and its TNF alpha and TNF beta complexes. Multiple receptor binding sites on TNF alpha trimers could also be demonstrated by a TNF alpha-induced agglutination of Latex beads coated with the rsTNFR beta-h gamma 3 fusion protein. Both rsTNFR beta and rsTNFR beta-h gamma 3 were found to inhibit binding of TNF alpha and TNF beta to native 55- and 75-kDa TNF receptors and to prevent TNF alpha and TNF beta bioactivity in a cellular cytotoxicity assay. Concentrations of rsTNFR beta-h gamma 3 equimolar to TNF alpha were sufficient to neutralize TNF activity almost completely, whereas a 10-100-fold excess of rsTNFR beta was needed for similar inhibitory effects. In view of their potent TNF antagonizing activity, recombinant soluble TNF receptor fragments might be useful as therapeutic agents in TNF-mediated disorders.

Animals

Recombinant soluble tumor necrosis factor receptor proteins protect mice from lipopolysaccharide-induced lethality.

The in vivo efficacy of human recombinant soluble tumor necrosis factor (TNF) receptor protein to prevent and to treat lipopolysaccharide (LPS)-induced lethal toxicity in D-galactosamine-treated mice was investigated. Chimeric proteins of the receptor extracellular domains fused to the hinge region of human IgG3 were expressed in myeloma cells (rsTNFR-h gamma 3). The fusion proteins had a disulfide-bonded dimeric structure. Upon intravenous injection, their serum concentration decreased relatively slowly after an initial phase of rapid elimination. D-galactosamine-sensitized mice were fully protected from the toxic effects of LPS, if the animal were pretreated with rsTNFR-h gamma 3 at 20 micrograms/animal. Partial protection was seen at significantly lower doses and when rsTNFR-h gamma 3 was given up to 3 h after LPS.

Animals

[Three cases of intramural cyst of the esophagus with ciliated epithelium].

We had 3 cases of intramural cyst of the esophagus with ciliated epithelium. In Case 1, 13-year-old man, the cyst was lined with ciliated columnar epithelium and had a smooth muscle layer. In Case 2, 37-year-old female, the cyst was lined with ciliated stratified epithelium and had some layers of smooth muscle. In Case 3, 42-year-old male, the cyst was lined with ciliated columnar epithelium and had a smooth muscle layer. In all cases, cartilaginous tissue was not found. They had no complaints and the abnormal shadow was found by chance Chest X-ray film. When the cyst did not have typical diagnostic criteria, clear division into esophageal cyst or bronchogenic cyst is difficult. The bronchus and the esophagus are of the foregut origin, so we suppose the middle type cyst, between esophageal and bronchial, may exist. In Case 2, the cyst was diagnosed as esophageal, because it had some muscle layers. But in Case 1 and Case 3, as the cyst did not have typical diagnostic criteria, we diagnosed them as the inclusion cyst of the esophagus, not as esophageal, not as bronchogenic.

Adolescent